043194   HUANG Q, MA B, SU Y, CHAN K, QU H, HUANG J, WANG D, QIU J, LIU H, YANG X, et al. (Sun Yat-sen Univ, Guangdong 510655, China, Email: yangxl28@mail.sysu.edu.cn) : MiR-197-3p represses the proliferation of prostate cancer by regulating the VDAC1/AKT/β-catenin Signaling Axis. Int J Biol Sci 2020, 16(8), 1417- 26.

Accumulating investigations have demonstrated that microRNAs (miRNAs) are promising efficient targets for the next generation of molecular therapeutics. The development of miRNA-based therapies requires the identification and validation of cancer-associated miRNAs. Herein, we identified that miR-197-3p regulates the carcinogenesis and development of prostate cancer (PCa) via bioinformatics analysis. Next, we investigated the function and regulatory mechanisms of miR-197-3p in PCa. Overexpression of miR-197-3p suppressed PCa cell proliferation and colony formation. In contrast, inhibition of miR-197-3p activity enhanced PCa cell proliferation and colony formation. Mechanistic investigations identified that voltage dependent anion channel 1 (VDAC1) is a direct target of miR-197-3p. miR-197-3p targeting of VDAC1 resulted in downregulation of p-Akt and β-catenin. Subsequently, we found that restoration of VDAC1 abolished the effects of miR-197-3p on PCa cell proliferation and AKT signaling pathway. Furthermore, we confirmed that miR-197-3p suppressed tumor xenograft growth in vivo. In conclusion, our study offers an empirical investigation of miR-197-3p, a tumor suppressor that may be a potential therapeutic target in PCa.

8 illus, 32 ref

045125   VAGHELA J F, ANAND T (Community Health Dep, St. Stephen’s Hospital, Sunder Nagari, Delhi) : Long-term follow-up of multidrug resistant tuberculosis patients of Delhi, who had received dots plus treatment along with home care and counseling. J Commun Dis 2020, 52(2), 50-6.

To realize the vision of a world free of TB, it becomes mandatory to disallow Ex-TB and Ex-MDR TB patients to have relapse or become Extremely Drug Resistant (XDR). Long follow-up of treated cases, who also received home care and counseling hasn’t been done thus far in India. Objective:To find out whether 50 % of Ex-MDR TB patients who had also received home based support with counseling are healthy after five years. This retrospective study of 109 Ex-MDR-TB patients was carried out by Community Health Department (CHD) of a Tertiary Care Hospital in Delhi. They had received daily MDR Regimen (starting from August 2009 to August 2010 from their respective DOTS Providers) and home care and strong counseling support. They completed it by 2011 or 2012. The study period was from May to August 2015.  After a long follow up of these patients in 2015 it was found that 71(65.14 %) were alive & healthy, 26(23.85 %) dead, 5(4.59 %) defaulters, 1(0.92 %) failure cases, 4(3.67 %) lost to follow-up, whereas 2(1.83 %) were on treatment as they had relapsed. Our study concludes that more than 50 % Ex-MDR TB cases who had also received home based support with counseling are alive & healthy after five years. We recommend that a mechanism for home-based care & counseling and Long Follow Up for MDR TB patients be developed.

6 tables, 20 ref

043193   DENG X, LIU J, LIU L, SUN X, HUANG J, DONG J (Integrative Medicine Dep, Fudan Univ, Shanghai 200040, China, Email: jcdong2004@126.com) : Drp1-mediated mitochondrial fission contributes to baicalein-induced apoptosis and autophagy in lung cancer via activation of AMPK signaling pathway. Int J Biol Sci 2020, 16(8), 1403-16.

Baicalein (BA), a natural compound extracted from Scutellaria baicalensis Georgi, has been reported to exert antitumor effect in various cancers. However, the underlying mechanisms have not been well demonstrated. In the present study, we focused on the relationship between mitochondrial fission and BA-induced apoptosis and autophagy. We showed that BA inhibited cell viability and induced mitochondrial apoptosis in A549 and H1299 lung cancer cells. BA induced the loss of mitochondrial membrane potential (MMP) and the release of cytochrome c and apoptosis inducing factor (Aif) from mitochondria to cytoplasm. Meanwhile, BA induced autophagy and activated autophagic flux. Furthermore, we found that BA induced mitochondrial fission and mitochondrial impairment. Blocking mitochondrial fission by mdivi-1 attenuated BA-induced apoptosis and autophagy. Moreover, BA activated AMP-activated protein kinase (AMPK) pathway. Knockdown of AMPK with lentivirus encoded AMPKα also attenuated BA-induced mitochondrial fission, apoptosis and autophagy. Our in vivo data confirmed that BA inhibited tumor growth and induced apoptosis and autophagy in a Lewis lung carcinoma (LLC) xenograft model via activation of AMPK/mitochondrial fission pathway. Our study highlights the critical role of AMPK/mitochondrial fission pathway in the regulation of BA-induced apoptosis and autophagy. These results revealed the molecular mechanism of the anti-lung cancer property of BA and provided novel perspectives for the application of BA in the treatment of lung cancer.

8 illus, 43 ref

045124   SINGH S K, SHARMA S N (National Centre for Disease Control, 22 Sham Nath Marg, Delhi) : Situational analysis and trend of covid-19 virus in India. J Commun Dis 2020, 52(2), 38-45.

COVID-19 virus has emerged a serious threat to the universe engulfing lakhs of human lives across the world. This is in the form of very unique and virulent deadly disaster on the earth spreading disease so fast being contagious in nature that it has given no time to medical professional to think and prepare to fight against this deadly disease. Loss of heavy lives in the world  can not be forgotten for long in the history as it has been reported from USA, Spain, Italy and France. It is said to have originated initially from Wuhan of China and spread over to other parts of the world. The point of spread or transmission has become a matter of debate and controversy around the world as to whether its transmission has happened  due to man to man contact after making shift from wild animals i.e. bats, pangolins (food market) or it was as a result of some leakage from a testing laboratory in Wuhan. Unfortunately, no drug or vaccine has been yet found effective to deal with this deadly virus. The spread of  this deadly virus is very rapid in the community due to its very high contagious nature and fast multiplication. There have been some updates in the knowledge about the clinical signs and symptoms of this virus across the world. However, the disease is changing its epidemiological profile with disease spectrum  and making difficult to deal with it. An attempt has been made in this manuscript to undertake the situational analysis and study the trend of COVID-19 in Indian perspective.

3 illus, 2 tables, 23 ref

043192   XU W, HUANG M, GUO J, ZHANG H, WANG D, LIU T, LIU H, CHEN S, GAO P, MU K (Pathology Dep, Shandong Univ, Wen Hua Xi Road 44, Jinan, 250012, China, Email: mukun@sdu.edu.cn) : The role of CHK1 varies with the status of oestrogenreceptor and progesterone-receptor in the targeted therapy for breast cancer. Int J Biol Sci 2020, 16(8), 1388- 402.

The therapeutic effects of the checkpoint kinase 1 (CHK1)-targeted inhibition in tumor therapy have been confirmed, but how to choose an effective application method in breast cancer with heterogeneous molecular characteristics has remained unclear. We evaluated the status of CHK1 in breast cancer using the cancer genome atlas database. Chemosensitivity and single-agent antitumor activity of CHK1 inhibition were measured by drug sensitivity assay, cell proliferation assay, cell cycle and apoptosis analysis in breast cancer with different ER/PR status. And based on the conjoint transcriptome atlas analyses, the corresponding mechanism were explored. In ER− /PR− /HER2− breast cancer, CHK1 inhibition enhanced adriamycin (ADR) chemosensitivity which was mediated by the mitotic checkpoint complex (MCC)–anaphase-promoting complex/cyclosome (APC/C)–cyclin B1 axis, Msh homeobox 2 (MSX2) and Bcl-2–like protein 11 (BIM). However, in ER+/PR+/HER2− breast cancer, because of the significant suppression for centromere protein F (CENPF)-mediated transcriptional activation of CHK1 induced by ADR itself, CHK1 inhibition fails to sensitize ADR toxicity. Interestingly, CHK1 inhibition showed the single-agent antitumor activity in ER+/PR+/HER2− breast cancer which was mediated by the cyclin dependent kinase inhibitor 1A (p21), kinesin family member 11 (Eg5) and cell surface death receptor (Fas). CHK1’s variable role determines the application of CHK1 inhibition in breast cancer with ER/PR heterogeneity.

7 illus, 44 ref

043191   LI L, HAN Z, QIU L, KANG D, ZHAN Z, TU H, CHEN J (Fujian Normal Univ, Fuzhou 350007, P. R. China, Email: lhli@fjnu.edu.cn) : Label-free multiphoton imaging to assess neoadjuvant therapy responses in breast carcinoma. Int J Biol Sci 2020, 16(8), 1376-87.

Neoadjuvant chemotherapy has been used increasingly in patients with early-stage or locally advanced breast carcinoma, and has been recommended as a general approach in locally advanced-stage diseases. Assessing therapy response could offer prognostic information to help determine subsequent nursing plan; particularly it is essential to identify responders and non-responders for the sake of helping develop follow-up treatment strategies. However, at present, diagnostic accuracy of preoperative clinical examination are still not satisfactory. Here we presented an alternate approach to monitor tumor and stroma changes associated with neoadjuvant therapy responses in breast carcinoma, with a great potential for becoming a new diagnostic tool—multiphoton microscopy. Imaging results showed that multiphoton imaging techniques have the ability to label-freely visualize tumor response such as tumor necrosis, and stromal response including fibrosis, mucinous response, inflammatory response as well as vascular hyperplasia in situ at cellular and subcellular levels. Moreover, using automated image analysis and a set of scoring methods, we found significant differences in the area of cell nucleus and in the content of collagen fibers between the pre-treatment and post-treatment breast carcinoma tissues. In summary, this study was conducted to pathologically evaluate the response of breast carcinoma to preoperative chemotherapy as well as to assess the efficacy of multiphoton microscopy in detecting these pathological changes, and experimental results demonstrated that this microscope may be a promising tool for label-free, real-time assessment of treatment response without the use of any exogenous contrast agents.

9 illus, 1 table, 36 ref

045123   RAJENDRAN R, REGU K, ANUSREE S B, RAJENDRAN A, JAIN S K, SINGH S K (National Centre for Disease Control, Calicut, Kerala) : The first covid-19 incidence in India: A lesson of struggle and survival. J Commun Dis 2020, 52(2), 25-31.

The coronavirus disease 2019 (COVID-19) outbreak, which originated in Wuhan, China, has now spread to more than 200 countries and administrative regions infecting 3,09,04,45 individuals of all ages as of 3rd April, 2020. Though most of the infected individuals exhibit mild symptoms including fever, upper respiratory tract infections, shortness of breath and diarrhea or are asymptomatic altogether. Severe cases of infection can lead to pneumonia, multiple organ failure and death. Globally, at least 2, 07,973 deaths have been directly attributed to COVID-19 and this number is expected to rise with the ongoing epidemic. WHO declared the outbreak to be a public health Emergency of International concern on January 30, 2020. The same day, a laboratory confirmed case of COVID-19 was reported in Kerala. That was the first reported case of COVID-19 in India. Since then 498 disease cases were reported in Kerala, while in India this has gone up to 33,050 with 1,074 deaths. During the first phase of the COVID-19 outbreak in Kerala, the health authorities have responded in a stellar manner. Kerala has not only traced hundreds of contacts of the confirmed cases and notified them to the Integrated Disease Surveillance Programme (IDSP) for monitoring, but also used unique community-based isolation methods, innovated while dealing with the Nipah virus outbreaks of 2018 and 2019. The model of monitoring with the District Collector as the administrative unit has been shared as a best practice with all states.

3 illus, 14 ref

045122   RAO A M K M, KANCHANA N D, SHARMA S N (WHO, Geneva) : Sampling model on surveillance of covid-19 virus among human populations. J Commun Dis 2020, 52(2), 12-7.

India stands at 90,927 confirmed cases of Covid-19 infection with WHO classification of clusters of cases. The governments around the world including India are mostly testing only people showing symptoms of flu, which are underestimating the real numbers due to presence of asymptomatic humans under virus incubation period. Random surveillance testing should, therefore, start as soon as possible to establish the mortality rate and virality of the Coronavirus. Such surveillance for Covid-19 provides valuable early warning information about the spread of the virus and also reduces the uncertainty surrounding the true extent of the pandemic and its mortality rate. Considered one of the most reliable forms of data collection, surveillance testing is takes small number of demographically representative random people to use as an estimate for the whole population. Due to these reasons, attempt was made to develop a model for Covid-19 virus surveillance using rapid antibody testing method for screening the human populations on disease prevalence, which can be followed by rRT-PCR testing. This screening method was test checked in virtual situation in Qutbullapur Municipality of Medak district, Telangana state, wherein three different areas – High risk, Moderate risk and Low risk, each having 3 km area were identified in the Municipality with 2118 houses and projections were made for virtual sampling in 4 replicated unit areas. The projection of cost analysis came as Rs. 11,900 for first round rapid sample collection from randomly selected houses in entire Municipality, which appeared as a feasible financial projection for a disease like Covid-19. Major town/cities can consider to adopt this as surveillance method for covid-19 virus.

2 tables, 6 ref

043190   MOROSI L, MATTEO C, CERUTI T, GIORDANO S, PONZO M, FRAPOLLI R, ZUCCHETTI M, DAVOLI E, D’INCALCI M, UBEZIO P (Istituto di Ricerche Farmacologiche Mario Negri IRCCS, M. Negri 2, 20156 Milano, Email: lavinia.morosi@marionegri.it) : Quantitative determination of niraparib and olaparib tumor distribution by mass spectrometry imaging. Int J Biol Sci 2020, 16(8), 1363-75.

Optimal intratumor distribution of an anticancer drug is fundamental to reach an active concentration in neoplastic cells, ensuring the therapeutic effect. Determination of drug concentration in tumor homogenates by LC-MS/MS gives important information about this issue but the spatial information gets lost. Targeted mass spectrometry imaging (MSI) has great potential to visualize drug distribution in the different areas of tumor sections, with good spatial resolution and superior specificity. MSI is rapidly evolving as a quantitative technique to measure the absolute drug concentration in each single pixel. Different inorganic nanoparticles were tested as matrices to visualize the PARP inhibitors (PARPi) niraparib and olaparib. Normalization by deuterated internal standard and a custom preprocessing pipeline were applied to achieve a reliable single pixel quantification of the two drugs in human ovarian tumors from treated mice. A quantitative method to visualize niraparib and olaparib in tumor tissue of treated mice was set up and validated regarding precision, accuracy, linearity, repeatability and limit of detection. The different tumor penetration of the two drugs was visualized by MSI and confirmed by LC-MS/MS, indicating the homogeneous distribution and higher tumor exposure reached by niraparib compared to olaparib. On the other hand, niraparib distribution was heterogeneous in an ovarian tumor model overexpressing the multidrug resistance protein P-gp, a possible cause of resistance to PARPi. The current work highlights for the first time quantitative distribution of PAPRi in tumor tissue. The different tumor distribution of niraparib and olaparib could have important clinical implications. These data confirm the validity of MSI for spatial quantitative measurement of drug distribution providing fundamental information for pharmacokinetic studies, drug discovery and the study of resistance mechanisms.

7 illus, 4 tables, 58 ref

043189   ZHANG P, LI Y, FU Y, HUANG L, LIU B, ZHANG L, SHAO X M, XIAO D (Basic Sciences Dep, Loma Linda Univ School of Medicine, Loma Linda, CA 92350, Email: Dxiao@llu.edu) : Inhibition of autophagy signaling via 3-methyladenine rescued nicotine-mediated cardiac pathological effects and heart dysfunctions. Int J Biol Sci 2020, 16(8), 1349-62.

Cigarette smoking is a well-established risk factor for myocardial infarction and sudden cardiac death. The deleterious effects are mainly due to nicotine, but the mechanisms involved and theranostics remain unclear. Thus, we tested the hypothesis that nicotine exposure increases the heart sensitivity to ischemia/reperfusion injury and dysfunction, which can be rescued by autophagy inhibitor. Nicotine or saline was administered to adult rats via subcutaneous osmotic minipumps in the absence or presence of an autophagy inhibitor, 3-methyladenine (3-MA). After 30 days of nicotine treatment, the rats underwent the cardiac ischemia/reperfusion (I/R) procedure and echocardiography analysis, and the heart tissues were isolated for molecular biological studies. Nicotine exposure increased I/R-induced cardiac injury and cardiac dysfunction as compared to the control. The levels of autophagy-related proteins including LC3 II, P62, Beclin1, and Atg5 were upregulated in the reperfused hearts isolated from nicotine-treated group. In addition, nicotine enhanced cardiac and plasma ROS production, and increased the phosphorylation of GSK3β (ser9) in the left ventricle tissues. Treatment with 3-MA abolished nicotine-mediated increase in the levels of autophagy-related proteins and phosphorylation of GSK3β, but had no effect on ROS production. Of importance, 3-MA ameliorated the augmented I/R-induced cardiac injury and dysfunction in the nicotine-treated group as compared to the control. Our results demonstrate that nicotine exposure enhances autophagy signaling pathway, resulting in development of ischemic-sensitive phenotype of heart. It suggests a potentially novel therapeutic strategy of autophagy inhibition for the treatment of ischemic heart disease.

8 illus, 55 ref

043188   GUA S, LIU Y, GAO L, XIAO F, SHEN J, XING S, YANG F, ZHANG W, SHI Q, LI Y, et al. (Cardiology Dep, The First Affiliated Hospital of Zhengzhou Univ, No.1 Jianshe East Road, Zhengzhou, China, Email: zlszzu@126.com) : TBC1D25 regulates cardiac remodeling through TAK1 signaling pathway. Int J Biol Sci 2020, 16(8), 1335-48.

Cardiac remodeling is a major early event of heart failure, which is regulated by multiple signaling pathways. Here, we demonstrate that TBC1D25 is upregulated during pathological cardiac remodeling. The aim of this study is to determine the role of TBC1D25 in cardiac remodeling and to illustrate the underlying molecular mechanism. Specifically, cardiac remodeling was induced in TBC1D25-KO mice and their wild-type control mice through partial transverse aortic constriction (TAC) of aortic arch. Knockout TBC1D25 exacerbated cardiac hypertrophy, fibrosis and dysfunction. Meanwhile, TBC1D25 overexpression in both H9C2 cells and NRCMs alleviate Angiotensin II-induced cardiomyocyte hypertrophy in vitro. Moreover, TBC1D25 deficiency increases the phosphorylation levels of TAK1 and its downstream molecular (JNK and p38), whereas overexpressed TBC1D25 inhibits phosphorylation of TAK1, JNK and p38. And TAK1 is the key molecule in this process. Furthermore, we demonstrated that TBC1D25 could directly interacts with TAK1 by immunoprecipitation assay and GST pull-down assay, and the interaction needs the amino acids from at least 138 to 226 in the C-terminal region of TBC1D25 and from 1 to 300 in the C-terminal region of TAK1. We conclude that TBC1D25 suppresses pathological cardiac remodeling via regulating TAK1-JNK/p38 signaling pathway, which suggests that TBC1D25 will likely become a promising therapeutic target for heart failure.

8 illus, 37 ref

043187   GAO L, YANG M, WEI Z, GU M, YANG L, BAI C, WU Y, LI G (Inner Mongolia Univ, Hohhot, 010070, China, Email: gpengli@imu.edu.cn) : MSTN mutant promotes myogenic differentiation by increasing demethylase TET1 expression via the SMAD2/SMAD3 pathway. Int J Biol Sci 2020, 16(8), 1324-34.

Myostatin (MSTN) is mostly expressed in skeletal muscle and plays crucial roles in the negative regulation of muscle mass development. The methylation and demethylation of myogenesis-specific genes are major regulatory factors in muscle satellite cell differentiation. The present study was designed to investigate the mechanism of myogenic differentiation regulated by MSTN mutation (MT) and the methylation/demethylation state of downstream genes. The results showed that, in the MSTN-/+ satellite cells, a higher myotube fusion index and a larger myotube length were observed compared to the wild type controls; the genes associated with myogenesis were all up-regulated compared to the WT controls. The methylation of the promoters and gene bodies of PAX3, PAX7, MyoD, and MyoG were all down-regulated, while the expression of the key demethylase TET1 was significantly promoted. ChIP-qPCR was used to demonstrate that the SMAD2/SMAD3 complex combined with the promoter of TET1 to inhibit the activity of TET1 promoter, indicating that MSTN may regulate TET1 via SMAD2/SMAD3. The overexpression of TET1 in wild type cells promoted myogenic differentiation, increased the myotube index, and reduced the methylation of the associated genes. On the contrary, the knockdown of TET1 in the MSTN mutant cells resulted in the opposite phenomena as in the overexpressed cells. In conclusion, the myostatin mutant showed an increased transcriptional activity of TET1, inducing higher levels of demethylation and improving the transcriptional activity levels of myogenic differentiation-associated genes. The binding of SMAD2/SMAD3 directly to the TET1 promoter region indicated that the MSTN mutant demethylated the myogenesis-specific genes by up-regulating TET1, which is directly controlled by SMAD2/SMAD3.

7 illus, 41 ref

043186   XIA P, ZHANG F, YUAN Y, CHEN C, HUANG Y, LI L, WANG E, GUO Q, YE Z (Anesthesiology Dep, Xiangya Hospital of Central South Univ, Changsha, 410078, Hunan Province, China, Email: yezhi523@csu.edu.cn) : ALDH 2 conferred neuroprotection on cerebral ischemic injury by alleviating mitochondria-related apoptosis through JNK/caspase-3 signing pathway. Int J Biol Sci 2020, 16(8), 1303-23.

Past studies have indicated that the dysregulation of Aldehyde dehydrogenase 2 (ALDH2) is related to the pathogenesis of acute stroke. However, the underlying mechanisms of ALDH2-mediated acute stroke are still not well understood. Thus, our study was designed to explore the influence of ALDH2 in acute stroke and determine whether its related mechanisms are involved in regulating mitochondria-associated apoptosis modulating JNK/caspase-3 pathway. In vitro analysis on the gain and loss of ALDH2 and JNK function were performed to explore its influence on OGD/R injury and relevant signaling pathways. Our findings suggested that ALDH2 expression was significantly down-regulated in rats suffering from acute stroke and also in primary cortical cultured neurons and PC12 cells upon OGD/R stimulation. ALDH2 overexpression markedly decreased infarct size and improved neurological outcomes. Furthermore, ALDH2 overexpression significantly suppressed stroke-induced mitochondria-associated apoptosis and inhibited p-JNK activation and p-JNK/caspase-3 complex formation. Similarly, in in vitro OGD/R models, ALDH2 reintroduction not only promoted cellular viability and moderated LDH release, but also inhibited mitochondria-related apoptosis. Moreover, JNK inhibition relieved OGD/R-induced cellular injury and apoptosis while JNK activation aggravated them. Furthermore, ALDH2 overexpression and JNK inhibition significantly reduced caspase-3 activation and transcription which was triggered by OGD/R damage. Caspase-3 activation and transcription also re-elevated during activation of JNK in ALDH2-reintroduced cells. Finally, ChIP assay revealed that p-JNK was bound to caspase-3 promoter. Collectively, ALDH2 overexpression led to a significant reduction in mitochondria-related apoptosis via JNK-mediated caspase-3 activation and transcription in both in vitro and in vivo cerebral ischemia models.

9 illus, 54 ref

043185   RAJAMANICKAM V, YAN T, XU S, HUI J, XU X, REN L, LIU Z, LIANG G, WANG O, WANG Y (Wenzhou Medical Univ, Wenzhou, Zhejiang, 325035, P. R. China, Email: yi.wang1122@wmu.edu.cn) : Selective targeting of the TLR4 co-receptor, MD2, prevents colon cancer growth and lung metastasis. Int J Biol Sci 2020, 16(7), 1288-301.

Toll-like receptor (TLR) signaling is an emerging pathway in tumor cell invasion and metastasis. Myeloid differentiation protein-2 (MD2) contributes to ligand recognition and activation of TLRs in response to exogenous microbial insults or endogenous agents. We hypothesized that blocking MD2 using a specific inhibitor would prevent TLR4-mediated inflammatory responses and metastatic cancer growth. Here, we report that a MD2 inhibitor, L6H21, inhibited migration and invasion of LPS-activated colon cancer CT26.WT cells. These activities were accompanied by inhibition of nuclear factor-κB (NF-κB) activation, and thereby inhibition of the production of pro-inflammatory cytokines and adhesive molecules in colon cancer cells. Furthermore, L6H21 inhibited CT26.WT metastasis to the lung in BALB/c mice as well as suppressed colitis-induced colon cancer induced by azoxymethane/dextran sulfate sodium (AOM/DSS). Taken together, our results demonstrated that L6H21 suppressed tumor invasion and metastasis through blocking TLR4-MD2/NF-κB signaling axis. These findings reveal that inhibition of MD2 may be an important target for the development of colon cancer therapies.

6 illus, 32 ref

043184   LIN W, ZHOU Q, WANG C-Q, ZHU L, BI C, ZHANG S, WANG X, JIN H (Medical Oncology Dep, Zhejiang Univ School of Medicine, Hangzhou 310016, Zhejiang, China, Email: zhouqiyin@zju.edu.cn) : LncRNAs regulate metabolism in cancer. Int J Biol Sci 2020, 16(7), 1194-206.

Metabolic reprogramming is a hallmark of cancer. Mammalian genome is characterized by pervasive transcription, generating abundant non-coding RNAs (ncRNAs). Long non-coding RNAs (lncRNAs) are freshly discovered functional ncRNAs exerting extensive regulatory impact through diverse mechanisms. Emerging studies have revealed widespread roles of lncRNAs in the regulation of various cellular activities, including metabolic pathways. In this review, we summarize the latest advances regarding the regulatory roles of lncRNAs in cancer metabolism, particularly their roles in mitochondrial function, glucose, glutamine, and lipid metabolism. Moreover, we discuss the clinical application and challenges of targeting lncRNAs in cancer metabolism. Understanding the complex and special behavior of lncRNAs will allow a better depiction of cancer metabolic networks and permit the development of lncRNA-based clinical therapies by targeting cancer metabolism.

3 illus, 122 ref

043184   PAN Z, HUANG Y, QIAN H, DU X, QIN W, LIU T (Shanghai Univ of Traditional Chinese Medicine, Shanghai, China, 200031, Email: liute1979@126.com) : Superparamagnetic iron oxide nanoparticles drive miR-485-5p inhibition in glioma stem cells by silencing Tie1 expression. Int J Biol Sci 2020, 16(7), 1274-87.

Gliomas are highly malignant nervous system tumours. Studies shown that cancer stem cells are one of the main reasons underlying recurrence, metastasis, and poor prognosis in glioma cases. Our previous studies have found that superparamagnetic iron oxide nanoparticles (SPIONs) can act as nucleic acid carriers to drive intracellular overexpression of these nucleic acids. In this study, CD44+/CD133+ glioma stem cells (HuGSCs) were first isolated from surgically resected tissues from patients. qPCR and western blot results showed that Tie1 expression in HuGSCs was significantly higher thanexpression in CD44-/CD133- glioma cells. Bioinformatic analysis and luciferase reporter assays showed that miR-485-5p binds to specific loci on the 3′-UTR of Tie1 mRNA to inhibit Tie1 expression. Subsequently, miR-485-5p/miR-mut and SPION complexes were transfected into HuGSCs. Transmission electron microscopy showed that a highly dense metallic electron cloud is present in HuGSCs. At the same time, in vivo and in vitro studies showed that miR-485-5p@SPIONs can significantly inhibit HuGSC proliferation, invasion, tumourigenicity, and angiogenesis. In-depth analysis showed that Tie1 interacts with neuronal growth factors such as FGF2, BDNF, GDNF, and GFAP. qPCR and western blot results showed that in miR-485-5p@SPIONs-HuGSCs, the expression levels of Tie1 and stem cell markers (Oct4, Sox2, Nanog, CD44, and CD133), and even FGF2, BDNF, GDNF, and GFAP were significantly lower than thelevels in the control group (miR-mut@SPIONs-HuGSCs). Therefore, this study showedthat Tie1 is an important factor that maintains glioma stem cell activity. SPIONs drive miR-485-5p overexpression in cells and inhibit endogenous Tie1 expression to downregulate the protein expression levels of Fgf2/GDNF/GFAP/BDNF and significantly weaken the in vivo and in vitro viability of gliomas.

7 illus, 37 ref

043183   CUI H, GUO S, HE H, GUO H, ZHANG Y, WANG B (Dermatology Dep, Shanxi Medical Univ, Taiyuan, Shanxi, China, Email: wbq_xy@sxent.org) : SASH1 promotes melanin synthesis and migration via suppression of TGF-β1 secretion in melanocytes resulting in pathologic hyperpigmentation. Int J Biol Sci 2020, 16(7), 1264-73.

Dyschromatosis universalis hereditaria (DUH) is an autosomal dominant pigmentary genodermatosis characterized by the presence of patches of hyperpigmentation and hypopigmented macules distributed over the body, with most cases reported in Asia. DUH is a heterogeneous disease and a small portion of patients carry the ABCB6 variant. In the present study, exome sequencing of four generations of a Chinese family with DUH identified a c.1761C>G (p.Ser587Arg) mutation in exon 15 of SAM and SH3 domain containing 1 (SASH1) that was found to co-segregate in some family members. Immunohistological analysis of biopsy specimens showed that SASH1 was diffusely distributed in all layers of the epidermis, suggesting increased transepithelial migration of melanocytes (MCs). The point mutation c.1761C>G of SASH1 was successfully induced in immortalized human melanocyte (PIG1) cells, which resulted in the downregulation of SASH1 expression. Bioinformatics analysis showed that mutated SASH1 downregulated thrombospondin 1 (THBS1) expression and inactivated transforming growth factor beta 1 (TGF-β1) signaling. TGF-β1 expression by PIG1cells was found to negatively regulate SASH1 protein expression. Transwell migration and wound-healing assays showed an increase in the migration and invasion capabilities of the cells carrying the mutation. Further, SASH1 mutations induced downregulation of melanin content. The study results suggest cross-talking between SASH1-TGF-β1 signaling, demonstrating the proposed MC migration modulation models and affecting melanin trafficking in the epithelium.

6 illus, 32 ref

043182   CHEN T-Q, HU N, HUO B, MASAU J F, YI X, ZHONG X-X, CHEN Y-J, GUO X, ZHU X-H, et al. (Huazhong Univ of Science and Technology, 1095 Jiefang Ave., Wuhan 430030, China, Email: jds@hust.edu.cn) : EHMT2/G9a inhibits aortic smooth muscle cell death by suppressing autophagy activation. Int J Biol Sci 2020, 16(7), 1252-63.

Although EHMT2 (also known as G9a) plays a critical role in several kinds of cancers and cardiac remodeling, its function in vascular smooth muscle cells (VSMCs) remains unknown. In the present study, we revealed a novel function of EHMT2 in regulating autophagic cell death (ACD) of VSMC. Inhibition of EHMT2 by BIX01294 or knockdown of EHMT2 resulted in reduced VSMC numbers which were independent of proliferation and apoptosis. Interestingly, EHMT2 protein levels were significantly decreased in VSMCs treated with autophagic inducers. Moreover, more autophagic vacuoles and accumulated LC3II were detected in VSMCs treated with BIX01294 or lenti-shEHMT2 than their counterparts. Furthermore, we found that EHMT2 inhibited the ACD of VSMCs by suppressing autophagosome formation. Mechanistically, the pro-autophagic effect elicited by EHMT2 inhibition was associated with SQSTM1 and BECN1 overexpression. Moreover, these detrimental effects were largely nullified by SQSTM1 or BECN1 knockdown. More importantly, similar results were observed in primary human aortic VSMCs. Overall, these findings suggest that EHMT2 functions as a crucial negative regulator of ACD via decreasing SQSTM1 or BECN1 expression and that EHMT2 could be a potent therapeutic target for cardiovascular diseases (e.g., aortic dissection).

6 illus, 44 ref

043181   QUIRICO L, ORSO F, ESPOSITO C L, BERTONE S, COPPO R, CONTI L, CATUOGNO S, CAVALLO F, FRANCISCIS V D, TAVERNA D (Torino Univ, Via Nizza, 52, 10126 Torino, Italy, Email: daniela.taverna@unito.it) : Axl-148b chimeric aptamers inhibit breast cancer and melanoma progression. Int J Biol Sci 2020, 16(7), 1238-51.

MicroRNAs (miRNAs) are small non-coding RNAs acting as negative regulators of gene expression and involved in tumor progression. We recently showed that inhibition of the pro-metastatic miR-214 and simultaneous overexpression of its downstream player, the anti-metastatic miR-148b, strongly reduced metastasis formation. To explore the therapeutic potential of miR-148b, we generated a conjugated molecule aimed to target miR-148b expression selectively to tumor cells. Precisely, we linked miR-148b to GL21.T, an aptamer able to specifically bind to AXL, an oncogenic tyrosine kinase receptor highly expressed on cancer cells. Axl-148b conjugate was able to inhibit migration and invasion of AXL-positive, but not AXL-negative, cancer cells, demonstrating high efficacy and selectivity in vitro. In parallel, expression of ALCAM and ITGA5, two miR-148b direct targets, was reduced. More importantly, axl-148b chimeric aptamers were able to inhibit formation and growth of 3D-mammospheres, to induce necrosis and apoptosis of treated xenotransplants, as well as to block breast cancer and melanoma dissemination and metastatization in mice. Relevantly, axl aptamer acted as specific delivery tool for miR-148b, but it also actively contributed to inhibit metastasis formation, together with miR-148b. In conclusion, our data show that axl-148b conjugate is able to inhibit tumor progression in an axl- and miR-148b-dependent manner, suggesting its potential development as therapeutic molecule.

6 illus, 35 ref

043180   MAO M, ZHANG A, HE Y, ZHANG L, LIU W, SONG Y, CHEN S, JIANG G, WANG X (Clinical Laboratory Dep, Sun Yat-sen Univ Cancer Center, Guangzhou 510060, Guangdong Province, China, Email: wangxuep@sysucc.org.cn.) : Development and validation of a novel nomogram to predict overall survival in gastric cancer with lymph node metastasis. Int J Biol Sci 2020, 16(7), 1230-7.

Gastric cancer (GC) with lymph node metastasis (LNM) at diagnosis is associated with a unstable prognosis and indefinite survival times. The aim of the present study was to construct and validate a model for the Overall survival (OS) estimation for patients with LNM. The nomogram was constructed to predict the OS for LNM-positive GC using the primary group of 836 patients and validated using an independent cohort of 411 patients. Factors in the nomogram were identified by multivariate Cox hazard analysis. The predictive capability of nomogram was evaluated by calibration analysis and decision curve analysis. Multivariate analysis suggested that eight pre-treatment characteristics were used for developing the nomogram. In the primary cohort, the C-index for OS prediction was 0.788 (95 % CI: 0.753-0.823), while in validation cohort, the C-index for OS prediction was 0.769 (95 % CI: 0. 720-0.818). The calibration plot for the probability of OS and decision curve analyses showed an optimal agreement. Based on the nomogram, we could divided patients into three groups: low-risk group, middle-risk group and a high-risk group(p <0.001).Taken together, we have provided an easy-to-used and accurate tool for predicting OS, furthermore could be used for risk stratification of OS of LNM-positive GC patients.

5 illus, 2 tables, 26 ref

046528   MALAR A, SURESH J, RAJAMANI K, BOOMIGA M (Medicinal and Aromatic Crops Dep, Tamilnadu Agricultural Univ, Coimbatore- 641 003, Email: boomigamani@gmail.com) : Effect of different growing media on growth and development of rooted cuttings of Indian sarsaparilla (Hemidesmus indicus L.). Med Plants 2020, 12 (1), 11-4.

Indian sarsaparilla (Hemidesmus indicus L.) is an important medicinal plant widely used in Indian systems of medicine as an antipyretic, blood purifier and refrigerant. It is under the list of endangered species in India due to the indiscriminate collection and utilization. To bring the plant under commercial cultivation, standardization of growing medium for rooted cuttings in Indian sarsaparilla was done at Department of Medicinal and Aromatic Crops, Horticultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. The rooted cuttings under mist chamber were transferred to PVC pipes measuring 5 feet length and 2 ½ inch diameter kept under open condition. The media like coir pith compost, vermicompost and red soil: sand: FYM (2:1:1) mixture and their combinations were used. Observations on root length, number of roots, fresh weight of the root, dry weight of the root, shoot length, number of laterals, number of leaves per plant, fresh weight of shoot and dry weight of shoot were recorded after 60 days of planting. Among the five treatments, growing medium M1 (coir pith compost) recorded longest root length (76.63 cm), more number of roots (67.25), maximum fresh weight of the roots (9.38 g), maximum dry weight of roots (1.73 g), highest shoot length (34.28 cm), more number of laterals (3.25), more number of leaves (42.25), maximum fresh weight of shoot (5.23 g) and maximum dry weight of shoots (1.25 g). Hence, from this study we can conclude that, use of coir pith compost medium provides good ancourage and high root yield to the Hemidesmus plants and thus can be utilized for commercial cultivation of the plants.

4 tables, 13 ref

043179   HOU J, ZHUO H, CHEN X, CHEN J, ZHENG W, ZHONG M, CAI J (Zhongshan Hospital affiliated to Xiamen Univ, Xiamen, China 361004, Email: caijianchun@xmu.edu.cn) : MiR-139-5p negatively regulates PMP22 to repress cell proliferation by targeting the NF-κB signaling pathway in gastric cancer. Int J Biol Sci 2020, 16(7), 1218-29.

Gastric cancer (GC) is one of the most common malignant tumors worldwide. Peripheral myelin protein 22 (PMP22) is a 22-kDa tetraspan glycoprotein that is predominantly expressed by myelinating Schwann cells. However, recent studies have shown that PMP22 is closely related to cell proliferation and tumorigenesis in different cancers. In this study, we discovered a new miRNA that regulates PMP22 and gastric cancer cell prolifration. Our bioinformatics analysis suggested that there is a conserved miRNA recognition site for miR-139-5p on the 3’ UTR of PMP22. Interestingly, our results showed overexpression of miR-139-5p significantly suppressed growth and prolifration in GC cells and inhibited tumor growth in nude mice xenografted with GC cells. MiR-139-5p suppressed the activity of a luciferase reporter containing the PMP22-3’ UTR, and the ectopic expression of PMP22 rescued the miR-139-5p-mediated inhibition of cell proliferation in GC cells. Mechanistically, miR-139-5p may negatively regulate PMP22 to repress cell proliferation by targeting the NF-κB signaling pathway in gastric cancer. Finally, overexpression of miR-139-5p significantly inhibited tumor growth in nude mice xenografted with GC cells.and the miR-139-5p levels were inversely correlated with PMP22 expression in nude mice tumor. Taken together, our data suggest an important regulatory role of miR-139-5p in gastric cancer, suggesting that miR-139-5p and PMP22 might be important diagnostic or therapeutic targets for gastric cancer and other human diseases.

7 illus, 2 tables, 43 ref

043178   CHEN H-D, HUANG C-S, XU Q-C, LI F, HUANG X-T, WANG J-Q, LI S-J, ZHAO W, YIN X-Y (Pancreato-Biliary Surgery Dep, The First Affiliated Hospital of Sun Yat-sen Univ, Guangzhou 510080, China, Email: yinxy@mail.sysu.edu.cn) : Therapeutic targeting of CDK7 suppresses tumor progression in intrahepatic cholangiocarcinoma. Int J Biol Sci 2020, 16(7), 1207-17.

Intrahepatic cholangiocarcinoma (ICC) is a lethal malignancy with high mortality and lack of effective therapeutic targets. Here, we found that expression of cyclin-dependent kinase 7 (CDK7) was significantly associated with higher tumor grade and worse prognosis in 96 ICC specimens. Depletion of CDK7 significantly inhibited cell growth, induced a G2/M cell cycle arrest, and reduced the migratory and invasive potential in ICC cells. Subsequent experiments demonstrated that ICC cells were highly sensitive to the CDK7 inhibitor THZ1. A low concentration of THZ1 markedly inhibited cell growth, cell cycle, migration, and invasion in ICC cell lines. RNA-sequencing (RNA-seq) analysis revealed that THZ1 treatment decreased the levels of massive oncogene transcripts, particularly those associated with cell cycle and cell migration. Quantitative reverse transcriptase PCR (qRT-PCR) analysis confirmed that transcription of oncogenes involved in cell cycle regulation (AURKA, AURKB, CDC25B, CDK1, CCNA2, and MKI67) and the c-Met pathway (c-Met, AKT1, PTK2, CRK, PDPK1, and ARF6) was selectively repressed by THZ1. In addition, THZ1 exhibited significant anti-tumor activity in a patient-derived xenograft (PDX) model of ICC, without causing detectable side effects.

7 illus, 1 table, 24 ref

046527   MINAEIAN S, MOHEBALI M, ELIKAEE S, AKHOUNDI B, SALEMI B, KHANALIHA K (Iran Univ of Medical Sciences, Tehran, Iran, Email: khanaliha.kh@iums.ac.ir) : Antileishmanial activity of Allium hirtifolium on promastigote and amastigote developmental stages of the parasite, Leishmania major. Med Plants 2020, 12(1), 5-10.

Cutaneous leishmaniasis is one of the most common parasitic diseases with great public health concern. The aim of this study is to investigate the effect of Allium hirtifolium on promastigote and amastigote developmental stages of Leishmania major. L. major promastigote was cultured in RPMI-1640 medium. The viability of promastigote stage after 24, 48, and 72 hr incubation using alcoholic extracts of A. hirtifolium and Meglumine evaluated by MTT Assay and IC₅₀ values (50 % inhibitory concentrations) were calculated. Amastigote growth in mouse macrophages was evaluated. The efficacy of different doses of the A. hirtifolium and Meglumine were calculated as 24, 48, and 72 hr after addition. The promastigote growth stage of L. major was inhibited by A. hirtifolium extracts after 24, 48 and 72 hours. The IC₅₀ of Allium hirtifolium extracts on amastigote growth of L. major were µg /ml,70.1 µg/ml and 51.4 µg/ml after 24, 48, and 72 hr respectively. Alcoholic extract of A. hirtifolium was reported to have anti-parasite effect in vitro on both the promastigote and amastigote development stages.

2 illus, 3 tables, 26 ref

043177   WANG D, QI H, ZHANG H, ZHOU W, LI Y, LI A, LIU Q, WANG Y (Shenzhen Univ, Shenzhen, Guangdong (518055), P.R.China, Email: yunw@szu.edu.cn.) : TAF1L promotes development of oral squamous cell carcinoma via decreasing autophagy-dependent apoptosis. Int J Biol Sci 2020, 16(7), 1180-93.

This study focused on investigating the relationships of TAF1L expression and clinical features or pathological stages of oral squamous cell carcinoma (OSCC), and its potential roles of TAF1L on OSCC development. Western blot and immunohistochemical staining were used to detect TAF1L expression in OSCC tissues and cells. Effects of TAF1L on OSCC cells in vitro were examined by cell proliferation assay, wound healing assay, transwell chamber assay, flow cytometry analysis and siRNA technique. Cellular key proteins related to cell autophagy and apoptosis were evaluated by Western blot and immunofluorescent staining. Moreover, functions of TAF1L on OSCC process were observed in nude mouse model. Testing results showed that expression of TAF1L protein was higher in OSCC tissues than that in normal oral epithelial or paracancerous tissues. Additionally, the level of TAF1L protein expression was upregulated in OSCC cell lines, compared to that in normal oral epithelial cells. Furthermore, cell proliferation, migration, autophagy and apoptosis were modulated post siRNA-TAF1L treatment in vitro. Especially, TAF1L knockdown-induced apoptotic activation on OSCC cells could be rescued by autophagic activator (Rapamycin). Moreover, that overexpression of TAF1L protein could promote the growth of OSCC cell xenografts was confirmed in nude mouse model. Taken together, it suggests that TAF1L may facilitate OSCC cells to escape cell apoptosis via autophagic activation for enhancing OSCC development.

7 illus, 3 tables, 46 ref

043176   ZHENG Y, CUI B, SUN W, WANG S, HUANG X, GAO H, GAO F, CHEN Q, LU L, AN Y, et al. (Physiology and Pathophysiology Dep, Fudan Univ, Shanghai 200032, China, Email: xbli@fudan.edu.cn) : Potential crosstalk between liver and extra-liver organs in mouse models of acute liver injury. Int J Biol Sci 2020, 16(7), 1166-79.

Carbon tetrachloride (CCl4), Concanavalin A (ConA), bile duct ligation (BDL), and liver resection (LR) are four types of commonly used mouse models of acute liver injury. However, these four models belong to different types of liver cell damage while their application situations are often confounded. In addition, the systematic changes of multiple extra-liver organs after acute liver injury and the crosstalk between liver and extra-liver organs remain unclear. Here, we aim to map the morphological, metabolomic and transcriptomic changes systematically after acute liver injury and search for the potential crosstalk between the liver and the extra-liver organs. Significant changes of transcriptome were observed in multiple extra-liver organs after different types of acute liver injury despite dramatic morphological damage only occurred in lung tissues of the ConA/BDL models and spleen tissues in the ConA model. Liver transcriptomic changes initiated the serum metabolomic alterations which correlated to transcriptomic variation in lung, kidney, and brain tissues of BDL and LR models. The potential crosstalk might lead to pulmonary damage and development of hepatorenal syndrome (HRS) and hepatic encephalopathy (HE) during liver injury. Serum derived from acute liver injury mice damaged alveolar epithelial cells and human podocytes in vitro. Our data indicated that different types of acute liver injury led to different transcriptomic changes within extra-liver organs. Integration of serum metabolomics and transcriptomics from multiple tissues can improve our understanding of acute liver injury and its effect on the other organs.

7 illus, 43 ref

046526   BABUJI A, INAMDAR S S (Pathology Dep, S. N. Medical Coll, Bagalkot, Karnataka, Email: abhijababuji.ab@gmail.com) : Haematological profile of dengue fever. Med Inn 2020, 9(1), 17-20.

Dengue is one of the most important viral diseases especially in the tropical regions. Dengue is caused by dengue virus (DEN) and is transmitted to humans by the bite of Aedes aegypti mosquito. Early clinical features of dengue infection are variable among patients, and initial symptoms are often nonspecific resembling any viral illness. Therefore, specific laboratory tests are necessary for an accurate diagnosis. To study the hematological profile of serologically diagnosed dengue patients in a tertiary care hospital. A total of 175 cases of dengue with serological confirmation of either dengue specific NS1 antigen assay and/or IgM and/or IgG antibodies were studied for the evaluation of haematological parameters from May 2019 to July 2019. Maximum patients 55 (31.4 %) were in age group of 21-30 years. A total leukocyte count of less than 4,000 cell/mm³ was present in 145 (82.85 %) patients. 26.28 % (46) of patients were having hematocrit >45 %. 47 (26.85 %) patients had mild thrombocytopenia; 105 (60 %) had moderate thrombocytopenia, and 18 (10.28 %) had severe thrombocytopenia. 21.14 % had a hemoglobin level of more than 15 g/dl. Haemoconcentration, leucopenia, thrombocytopenia gives clues to test for dengue serology so that dengue cases can be diagnosed in their initial stages as these cases does not have specific medical therapy, hence clinical recovery monitoring is largely dependent on haematological parameters.

6 tables, 7 ref

046525   DORLE A S, MANNAPUR B S, KULKARNI P, DESAI S, MUDARADDI R (Community Medicine Dep, S. N. Medical Coll, Bagalkot, Karnataka, Email: drmannapur15@gmail.com) : Effect of health education and IFA and albendazole on anemia among adolescent girls in rural field practice area of Bagalkot: A non-randomized interventional trial without control. Med Inn 2020, 9(1), 11-6.

Anemia is one of the most important public health problem among adolescent girls in developing countries. The incidence of anemia is more in rural areas. Health education plays a very important role in changing the food habits and behavior. To determine the prevalence of anemia & its influencing socio demographic factors and the effectiveness of intervention (health education and IFA & Albendezole) among adolescent girls. A Non-randomized interventional trial without control was conducted in the rural field practice area. The study participants were in the age group of 14-19 years. Pre-test was conducted to know the existing knowledge about anemia and a pre-test Hb % estimation was done. Health education sessions were conducted in the schools by peer educators, link workers and the staff. Anemic adolescent girls were made to take 100 IFA tablets and one albendazole tablet. Post-test evaluation survey was done to know the effectiveness of health education sessions and Iron and Folic Acid(IFA) Supplementation and albendazole tablets. Prevalence of anemia in adolescent girls was 53.18 %. The association between socio-economic status and anemia was found to be significant. There was significant increase in level of knowledge among adolescent girls regarding anemia after health education intervention and there was significant increase in Hemoglobin level among adolescent girls after IFA and Albendazole ingestion. The intensive health education and IFA and albendazole administration brings down the anemia problem which can prevent the anemia related complications in the future.

9 tables, 20 ref

046524   MUGHAL M B, SHAH S T M, FAROOQ M (General surgery Dep, Univ Hospital Hairmyres, Eaglesham Road, East Kilbride, Email: drmbm@yahoo.com) : Experience with mesh free tissue hernioplasty for primary inguinal hernia. Med Inn 2020, 9(1), 7-10.

Mesh-free tissue hernioplasty technique by Desarda has given a new trend in inguinal hernia treatment. Least complication rate simplicity, reproducibility, and cost- effectiveness are major hallmarks. This longitudinal study was conducted in a peripheral public hospital from December 2016 to December 2017. Eighty-one patients of primary inguinal hernia were included. Post-operative follow-up was done on the 4th and 8th day, then at one, three, six, twelve and twenty-four months. Post-operative assessment was done and early and late complications were noted. Eighty-one patients of primary inguinal hernia, all men, forty-nine right sided and thirty-two left sided, nine direct and seventy two indirect hernias, mean age was 37.9 years. All patients were able to be discharged on first post-operative day. There was no case of surgical site infection. Only one recurrence and no patient had chronic groin pain. This physiological mesh free tissue hernioplasty is a promising technique for primary inguinal hernia repair. It is simple, reproducible and cost effective. 

4 tables, 15 ref

046523   RAYKAR R R (Neurosurgery Dep, St. John Medical Coll and Hospital, Bangalore, Karnataka, Email: rajeshraykar1@gmail.com) : Clinical profile and surgical outcome of spinal tumors at a tertiary care hospital. Med Inn 2020, 9(1), 1-6.

Published literature on spinal cord tumors is relatively lesser compared to tumors of central nervous system. In case of spinal tumors, early diagnosis followed by surgery can bring back almost all functions of affected compared to other body organs, To study clinical profile and surgical outcome of spinal tumors. : A hospital based follow up study was carried out among 55 eligible cases of spinal tumors for three years. After recruitment, patients were operated and followed for six months at six weeks, three months and six months. At recruitment, and during follow up detailed history, thorough clinical examination was done. Samples for histopathology were sent after surgery.  Females (54.5 %) were more than males (45.5 %). Most commonly affected age group was 40-49 years (25.5 %). Most common location of spinal tumors was dorsal (50.9 %). Pain was most common presenting symptoms (74.5 %). Total resection was performed in 61.8 % cases. Schwannoma was most common histopathology type (32.7 %) followed by meningioma (21.8 %). Only three cases developed complications immediately after surgery and among them; two improved at follow-up. One patient was incontinent post-operatively and continued to be same at 3 months follow-up and became continent at 6 months. One case developed urinary incontinence at 3 months and continued to be same. Overall 51 cases (85 %) were absolutely normal from surgery to 6 months. 40-49 years of age people are commonly affected. It occurs more dorsally and presents commonly with pain. Surgical resection was successful in more than 90 % of the cases. 

4 tables, 15 ref

043175   BAI Y, WEI C, ZHONG Y, ZHANG Y, LONG J, HUANG S, XIE F, TIAN Y, WANG X, et al. (Immunology Dep, Capital Medical Univ, Beijing, China, Email: xiwang@ccmu.edu.cn) : Development and validation of a prognostic nomogram for gastric cancer based on DNA methylation-driven differentially expressed genes. Int J Biol Sci 2020, 16(7), 1153-65.

The incidence of gastric cancer (GC) ranks fifth among common tumors and GC is the third leading cause of cancer-related death worldwide. The aim of this study was to develop and validate a nomogram for predicting the overall survival (OS) of patients with GC. DNA methylation (DNAm)-driven genes were identified by integrating DNAm and gene expression profiling analyses from The Cancer Genome Atlas (TCGA) GC cohort. Then, a risk score model was built based on Kaplan-Meier (K-M), least absolute shrinkage and selector operation (LASSO), and multivariate Cox regression analyses. After analyzing the clinical parameters, a nomogram was constructed and assessed. Another cohort (GSE62254) was used for external validation. Thirteen differentially expressed DNAm-driven genes were narrowed down to a six-gene signature (PODN, NPY, MICU3, TUBB6 and RHOJ were hypermethylated, and MYO1A was hypomethylated), which was associated with OS (P < 0.05) after survival and LASSO regression analyses. These differentially expressed genes (DEGs) with altered DNAm statuses were included in the prognostic risk score model. The univariate Cox regression analysis indicated that risk score, age, and number of positive lymph nodes were significantly associated with survival time in GC patients. The multivariate Cox regression analysis also indicated that these variables were significant prognostic factors for GC. A nomogram including these variables was constructed, and its performance in predicting the 1-, 3- and 5-year survival outcomes of GC patients was estimated through time-dependent receiver operating characteristic (ROC) curves. In addition, the clinical benefit of this model was revealed by decision curve analysis (DCA). Pathway enrichment analysis suggested that these DNAm-driven genes might impact tumor progression by affecting signaling pathways such as the “ECM RECEPTOR INTERACTION” and “DNA REPLICATION” pathways. The altered status of the DNAm-driven gene signature (PODN, MYO1A, NPY, MICU3, TUBB6 and RHOJ) was significantly associated with the OS of GC patients. A nomogram incorporating risk score, age and number of positive lymph nodes can be conveniently used to facilitate the individualized prediction of OS in patients with GC.

7 illus, 44 ref

043174   PISA D, ALONSO R, CARRASCO L (Autónoma de Madrid Univ, Cantoblanco. 28049 Madrid. Spain, Email: mailto:lcarrasco@cbm.csic.es) : Parkinson’s disease: A comprehensive analysis of fungi and bacteria in brain tissue. Int J Biol Sci 2020, 16(7), 1135-52.

Parkinson’s disease (PD) is characterized by motor disorders and the destruction of dopaminergic neurons in the substantia nigra pars compacta. In addition to motor disability, many patients with PD present a spectrum of clinical symptoms, including cognitive decline, psychiatric alterations, loss of smell and bladder dysfunction, among others. Neuroinflammation is one of the most salient features of PD, but the nature of the trigger remains unknown. A plausible mechanism to explain inflammation and the range of clinical symptoms in these patients is the presence of systemic microbial infection. Accordingly, the present study provides extensive evidence for the existence of mixed microbial infections in the central nervous system (CNS) of patients with PD. Assessment of CNS sections by immunohistochemistry using specific antibodies revealed the presence of both fungi and bacteria. Moreover, different regions of the CNS were positive for a variety of microbial morphologies, suggesting infection by a number of microorganisms. Identification of specific fungal and bacterial species in different CNS regions from six PD patients was accomplished using nested PCR analysis and next-generation sequencing, providing compelling evidence of polymicrobial infections in the CNS of PD. Most of the fungal species identified belong to the genera Botrytis, Candida, Fusarium and Malassezia. Some relevant bacterial genera were Streptococcus and Pseudomonas, with most bacterial species belonging to the phyla Actinobacteria and Proteobacteria. Interestingly, we noted similarities and differences between the microbiota present in the CNS of patients with PD and that in other neurodegenerative diseases. Overall, our observations lend strong support to the concept that mixed microbial infections contribute to or are a risk factor for the neuropathology of PD. Importantly, these results provide the basis for effective treatments of this disease using already approved and safe antimicrobial therapeutics.

10 illus, 2 tables, 87 ref

043173   LU X, YANG F, CHEN D, ZHAO Q, CHEN D, PING H, XING N (Urology Dep, Capital Medical Univ, Beijing, China, Email: xingnianzeng@126.com) : Quercetin reverses docetaxel resistance in prostate cancer via androgen receptor and PI3K/Akt signaling pathways. Int J Biol Sci 2020, 16(7), 1121-34.

Docetaxel is the first-line chemotherapy agent for metastatic prostate cancer. However, the emergence of resistance diminishes its efficacy and limits the survival benefit. Quercetin is a dietary flavonoid which has been shown to have multiple anti-cancer effects. Also, quercetin has been reported to reverse chemo-resistance in many other cancers. This study was to determine whether quercetin could reverse docetaxel resistance in prostate cancer cells and xenograft models, thereby exploring the underlying mechanism. Depending on the docetaxel-resistant cells (LNCaP/R, PC-3/R) which were established from docetaxel-sensitive cells (LNCaP, PC-3), it was demonstrated that quercetin could reverse docetaxel resistance in prostate cancer on proliferation, colony formation, migration, invasion and apoptosis. Although single docetaxel application had little effect on docetaxel-resistant cells, combining docetaxel with quercetin was significantly effective. Combination therapy could maximally inhibited PI3K/Akt pathway and promoted apoptosis. As shown by in-vivo study, xenograft tumors treated by docetaxel with quercetin had poorest growth. Then, to investigate the underlying mechanisms, the differences among parental cells, docetaxel-resistant subclones and quercetin treated resistant subclones were evaluated. It was found that docetaxel-resistant subclones had stronger activation of androgen receptor and PI3K/Akt pathway, more remarkable mesenchymal and stem-like cell phenotypes, and more P-gp expression than that of parental cells. Interestingly, quercetin could reverse these transformations. Our data revealed that quercetin had docetaxel-resistance reversal effect both in vitro and in vivo and provided in-depth support for clinical use of quercetin in docetaxel-resistant prostate cancer.

8 illus, 32 ref

043172   LI H, ZHANG C, YANG C, BLEVINS M, NORRIES D, ZHAO R, HUANG M (Dermatology Dep, Colorado Anschutz Medical Campus Univ, Aurora CO 80045, USA, Email: mingxia.huang@cuanschutz.edu) : C-terminal binding proteins 1 and 2 in traumatic brain injury-induced inflammation and their inhibition as an approach for anti-inflammatory treatment. Int J Biol Sci 2020, 16(6), 1107-20.

Traumatic brain injury (TBI) induces an acute inflammatory response in the central nervous system that involves both resident and peripheral immune cells. The ensuing chronic neuroinflammation causes cell death and tissue damage and may contribute to neurodegeneration. The molecular mechanisms involved in the maintenance of this chronic inflammation state remain underexplored. C-terminal binding protein (CtBP) 1 and 2 are transcriptional coregulators that repress diverse cellular processes. Unexpectedly, we find that the CtBPs can transactivate a common set of proinflammatory genes both in lipopolysaccharide-activated microglia, astrocytes and macrophages, and in a mouse model of the mild form of TBI. We also find that the expression of these genes is markedly enhanced by a single mild injury in both brain and peripheral blood leukocytes in a severity- and time-dependent manner. Moreover, we were able to demonstrate that specific inhibitors of the CtBPs effectively suppress the expression of the CtBP target genes and thus improve neurological outcome in mice receiving single and repeated mild TBIs. This discovery suggests new avenues for therapeutic modulation of the inflammatory response to brain injury.

6 illus, 63 ref

043171   BIAN L, MENG Y, ZHANG M, GUO Z, LIU F, ZHANG W, KE X, SU Y, WANG M, YAO Y, et al. (Radiation Oncology Dep, Shanghai Jiaotong Univ School of Medicine, Shanghai, China, Email: lidong@shsmu.edu.cn) : ATM expression is elevated in established radiation-resistant breast cancer cells and improves DNA repair efficiency. Int J Biol Sci 2020, 16(6), 1096-106.

Repair of damaged DNA induced by radiation plays an important role in the development of radioresistance, which greatly restricts patients’ benefit from radiotherapy. However, the relation between radioresistance development and DNA double-strand break repair pathways (mainly non-homologous end joining and homologous recombination) and how these pathways contribute to radioresistance are unclear. Here, we established a radioresistant breast cancer cell line by repeated ionizing radiation and studied the alteration in DNA repair capacity. Compared with parental sham-treated cells, radioresistant breast cancer cells present elevated radioresistance, enhanced malignancy, increased expression of Ataxia-telangiectasia mutated (ATM), and increased DNA damage repair efficiency, as reflected by accelerated γ-H2AX kinetic. These defects can be reversed by ATM inhibition or ATM knockdown, indicating a potential link between ATM, DNA repair pathway and radiosensitivity. We propose that cancer cells develop elevated radioresistance through enhanced DNA damage repair efficiency mediated by increased ATM expression. Our work might provide a new evidence supporting the potential of ATM as a potential target of cancer therapy.

6 illus, 41 ref

043170   WANG H, WANG B, WEI J, MENG L, ZHANG Q, QU C, XIN Y, JIANG X (Jilin Univ, 126 Xinmin Street, Changchun, 130021 China, Email: xiny@jlu.edu.cn) : Molecular mechanisms underlying increased radiosensitivity in human papillomavirus-associated oropharyngeal squamous cell carcinoma. Int J Biol Sci 2020, 16(6), 1035-43.

Oropharyngeal squamous cell carcinoma (OPSCC) is an important type of head and neck squamous cell carcinoma (HNSCC). The traditional risk factors for OPSCC include carcinogen intake, smoking, alcohol consumption, and lifestyle. In recent years, cases of human papillomavirus (HPV)-related OPSCC have gradually increased. At present, HPV-related OPSCC in developed Western countries comprise up to 90 % of all OPSCC cases, while in other developing countries, the proportion of HPV-related OPSCC cases is also gradually increasing. Compared with HPV-negative OPSCC, HPV-positive OPSCC patients have better overall survival rates and local control rates and this improved prognosis may be related to the increased radiosensitivity of HPV-positive tumors. Due to this more favorable prognosis, many downgraded treatment schemes are gradually emerging, including simple radiotherapy instead of concurrent radiotherapy or reduced radiotherapy dose. However, there is insufficient theoretical basis for such schemes. Some studies have shown that delayed repair of DNA damage after radiation, G2/M arrest, increased hypoxia, and decreased proliferation capacity are the main reasons for the increased radiosensitivity of HPV-positive tumor cells. In this review, we discuss the four principles of tumor cell damage caused by radiation, including repair, reoxygenation, redistribution, and regeneration in order to reveal the mechanism whereby HPV increases the radiosensitivity of tumor cells. An attempt was made to provide sufficient information to facilitate more individualized treatment for HPV-positive OPSCC patients, under the premise of good tumor control.

1 illus, 1 tables, 83 ref

043169   CHANG R-Q, ZHOU W-J, LI D-J, LI M-Q (Fudan Univ Shanghai Medical Coll, Shanghai 200082, People’s Republic of China, Email: djli@shmu.edu.cn) : Innate lymphoid cells at the maternal-fetal interface in human pregnancy. Int J Biol Sci 2020, 16(6), 957-69.

Pregnancy constitutes a major challenge to the maternal immune system, which must tolerate fetal alloantigen encoded by paternal genes. In addition to their role in inducing maternal-fetal immune tolerance, accumulating evidence indicates that decidual immune cells are involved in several processes required for a successful pregnancy, including trophoblast invasion as well as tissue and spiral artery remodeling. Innate lymphoid cells (ILCs), an important branch of the innate immune system, which has expanded rapidly in recent years, are strong actors in mucosal immunity, tissue homeostasis and metabolism regulation. With the recent identification of ILCs in the human decidua, the role of ILCs at the maternal-fetal interface raises concern. Herein, we review the presence and characterization of ILCs in the human decidua, as well as their function in normal pregnancy and pathological pregnancy, including reproductive failure, preeclampsia and others.

4 illus, 1 table, 171 ref

043168   WANG W, WANG C, XU H, GAO Y (Hepatology Dep, The First Hospital of Jilin Univ, Changchun, Jilin, 130021, China, Email: yanhang@mail.jlu.edu.cn.) : Aldehyde dehydrogenase, liver disease and cancer. Int J Biol Sci 2020, 16(6), 921-34.

Acetaldehyde dehydrogenase 2 (ALDH2) is the key enzyme responsible for metabolism of the alcohol metabolite acetaldehyde in the liver. In addition to conversion of the acetaldehyde molecule, ALDH is also involved in other cellular functions. Recently, many studies have investigated the involvement of ALDH expression in viral hepatitis, alcoholic liver disease (ALD), non-alcoholic fatty liver disease (NAFLD), liver fibrosis, and liver cancer. Notably, ALDH2 expression has been linked with liver cancer risk, as well as pathogenesis and prognosis, and has emerged as a promising therapeutic target. Of note, approximately 8 % of the world’s population, and approximately 30-40 % of the population in East Asia carry an inactive ALDH2 gene. This review summarizes new progress in understanding tissue-specific acetaldehyde metabolism by ALDH2 as well as the association of ALDH2 gene polymorphisms with liver disease and cancer. New research directions emerging in the field are also briefly discussed.

3 illus, 1 table, 123 ref

043167   XIAO X, ZHUANG X, XU C, CHEN H, ZHU W, PANG C P, ZHANG M (Shantou Univ, Shantou, China 515041, Email: lijingyu19801980@163.com) : ROBO4 deletion ameliorates PAF-mediated skin inflammation via regulating the mRNA translation efficiency of LPCAT1/LPCAT2 and the expression of PAF receptor. Int J Biol Sci 2020, 16(6), 1086-95.

The diminished level of platelet-activating factor acetylhydrolase (PAFAH) in milk causes an enhanced level of platelet activating factor (PAF) in the skin, leading to a severe hair loss phenotype during neonatal pup’s lactation. The deletion of very-low-density-lipoprotein receptor (VLDLR) prevents the expression and secretion of PAFAH. Here we revealed that deletion of Roundabout 4 (ROBO4) in mice ameliorated hair loss phenotype via reducing PAF concentration in skin. As a consequence, the neonatal pups with ROBO4 deletion lactated by mother with VLDLR deletion showed normal hair phenotype during lactation. In details,ROBO4 deletion reduced the protein but not mRNA expression of two PAF synthetic enzymes LPCAT1/LPCAT2 in macrophage as well as the expression of PAF receptor in both macrophage and ocular tissue, but increased PAFAH protein in serum. On the other hand, RNA expression profile analysis in macrophages revealed that the genes involving in oxidative phosphorylation and ribosome obviously decreased their expression in response to ROBO4 deletion. Moreover, through High Performance Liquid Chromatography (HPLC) analysis, we found that ATP concentration also reduced in ROBO4 deletion macrophages. Because ribosome and energy are very important factors for the mRNA translation, we then tested whether ROBO4 deletion affects LPCAT1/LPCAT2 mRNA translation using polyribosome assay. As expected, the mRNA level of LPCAT1/LPCAT2 significantly decreased in polyribosome in ROBO4 deletion macrophage comparing to that of wild type. Additionally, mice with ROBO4 deletion suppressed LPS-induced IL-6 expression as well as the phosphorylation of p44/42 and p65, but enhanced the AKT phosphorylation. Collectively, ROBO4 deletion alleviates PAF- and LPS-mediated inflammation. And above results also indicate PAF signal might be a crosstalk point of ROBO4- and VLDLR-activated pathways.

4 illus, 1 table, 21 ref

043166   LU W, YANG C, HE H, LIU H (Integrated Traditional and Western Medicine Dep, West China Hospital of Sichuan Univ, Chengdu 610041, Sichuan, China, Email: liuhong1980@scu.edu.cn) : The CARM1-p300-c-Myc-Max (CPCM) transcriptional complex regulates the expression of CUL4A/4B and affects the stability of CRL4 E3 ligases in colorectal cancer. Int J Biol Sci 2020, 16(6), 1071-85.

The transcription factor c-Myc and two cullin family members CUL4A/4B function as oncogenes in colorectal cancer. Our recent publication reveals that c-Myc specifically activates the expression of CUL4A/4B through binding to their promoters. However, the underlying mechanism of how c-Myc actions in this process is still unknown. Using mass spectrometry and immunoprecipitation assays, we identified c-Myc formed a transcriptional complex with its partner Max (Myc-associated factor X), a histone acetyltransferase p300 and a coactivator associated arginine methyltransferase 1 (CARM1) in the present study. Knockdown or overexpression of the components of CARM1-p300-c-Myc-Max (CPCM) complex resulted in a decrease or increase of CUL4A/4B levels, respectively. Individual knockdown or inhibition of CPCM components decreased cell proliferation, colony formation, and cell invasion. Biochemically, knockdown or inhibition of CPCM components decreased their occupancies on the promoters of CUL4A/4B and resulted in their downregulation. Importantly, inhibition of CPCM components also caused a decrease of CRL4 E3 ligase activities and eventually led to an accumulation of ST7 (suppression of tumorigenicity 7), the specific substrate of CRL4 E3 ligases in colorectal cancer. Moreover, the in vivo tumor formation results indicated that knockdown or inhibition of CPCM components significantly decreased the tumor volumes. Together, our results suggest that the CPCM complex mediates explicitly the expression of CUL4A/4B, and thus affects the stability of CRL4 E3 ligases and the ubiquitination of ST7. These results provide more options by targeting the CPCM components to inhibit tumor growth in the therapy of colorectal cancer.

9 illus, 61 ref

043165   YANG C, WU J, HE H, LIU H (Integrated Traditional and Western Medicine Dep, West China Hospital of Sichuan Univ, Chengdu 610041, Sichuan, China, Email: liuhong1980@scu.edu.cn) : Small molecule NSC1892 targets the CUL4A/4B-DDB1 interactions and causes impairment of CRL4^DCAF4 E3 ligases to inhibit colorectal cancer cell growth. Int J Biol Sci 2020, 16(6), 1059-70.

Cullin 4A and 4B (CUL4A and 4B) function as oncogenes in colorectal cancer (CRC) cells. Both of them conservatively associate with DNA damage–binding protein 1 (DDB1) and DDB1– CUL4-associated factor 4 (DCAF4) to form Cullin-RING E3 ligases known as CRL4^DCAF4, which specifically ubiquitinate and degrade tumor suppressor ST7 (suppression of tumorigenicity 7). Knockdown either CUL4A/4B or DDB1 significantly inhibits tumor cell growth in vitro and in vivo. Thus, targeting these CRL4^DCAF4 components and their interactions may be an effective strategy for the therapy of CRC. In this study, we developed an in vitro AlphaScreen assay to identify small molecules targeting the CUL4A-DDB1 interaction. We obtained a compound NSC1892, which strongly disrupted the CUL4A-DDB1 interaction (IC50 = 1.8 μM). Oncogenic phenotype analyses indicated that NSC1892 showed significant cytotoxicity to decrease cell proliferation, colony formation and invasion in CRC cells. Biochemical analyses demonstrated that NSC1892 treatment did not change CUL4A and CUL4B protein levels, but caused the degradation of DDB1, thereby leading to the impaired assembly of CRL4^DCAF4 E3 ligases and resulting in the accumulation of ST7. The administration of NSC1892 in mice also significantly inhibited tumor growth through degrading DDB1 and accumulating ST7. Interestingly, NSC1892 also showed promising cytotoxicity to decrease the growth of other CUL4A- or CUL4B-overexpressing tumor cells such as SKOV3 ovarian cells and Saos2 osteosarcoma cells. Our results provide a new avenue for the development of a therapeutic compound targeting tumors through disrupting the CUL4-DDB1 interaction.

7 illus, 26 ref

043164   GAO Q, LEI F, ZENG Q, GAO Z, NIU P, JUNNAN, NING, LI J, ZHANG J (General Surgery Dep, Capital Medical Univ, Beijing 100050. China, Email: zhangjun5986@ccmu.edu.cn) : Functional passenger-strand miRNAs in exosomes derived from human colon cancer cells and their heterogeneous paracrine effects. Int J Biol Sci 2020, 16(6), 1044-58.

Exosome-mediated microRNAs (miRNAs) are closely related to the occurrence, development, invasion, metastasis, therapeutic resistance, diagnosis and treatment of malignant tumors. Guide-strand miRNA and passenger-strand miRNA (miRNA*) exist in miRNA processing, but the function of passenger-strand miRNA is often overlooked. In this study, we attempted to identify functional miRNA*s in exosomes derived from human colon cancer SW620 cells. miRNA expression profiles of human normal colonic epithelial cells NCM460 and colon cancer cells SW620 were compared by high-throughput sequencing. According to the sequencing results, we defined two sets of differentially expressed miRNAs: “high in exosome and high in cell” (HEHC) and “high in exosome but low in cell” (HELC). As passenger-strand miRNAs, miR-2277-3p and miR-26b-3p, which belong to different sets, have diametrically opposite functions. MiR-2277-3p promotes proliferation, migration, and invasion of SW620 cells by targeting NUPR1L, while miR-26b-3p exerts an inhibitory effect by targeting PFDN1. Using exosomes as transport vectors, the effect of exosomes rich in miR-2277-3p on cells is consistent with the effect of liposome-transfected overexpressed miR-2277-3p, resulting in a cancer-promoting effect. However, exosomes rich in miR-26b-3p did not have a tumor suppressor effect. Further analysis revealed that exosomes rich in miR-2277-3p also had a high abundance of integrin β4. Altering the abundance of integrin β4 in exosomes changes the ability of exosomes to be taken up by cells, thereby altering the paracrine effects of exosomes. In summary, we revealed the fact that a large number of passenger-strand miRNAs exist in exosomes of colon cancer cells, these miRNAs are preliminarily categorized into two sets, and miR-2277-3p and miR-26b-3p, as representatives of each set, showed opposite functions. In addition, we revealed that integrin β4 is a marker of exosome heterogeneity in colon cancer cells, which directly correlates with the ability of exosomes to be uptaken by cells of the same kind, thus regulating the paracrine effect of exosomes.

10 illus, 31 ref

043163   LIU Q, SONG J, PAN Y, SHI D, YANG C, WANG S, XIONG B (Hubei Cancer Clinical Study Center, Wuhan 430071, China, Email: binxiong1961@whu.edu.cn) : Wnt5a/CaMKII/ERK/CCL2 axis is required for tumor-associated macrophages to promote colorectal cancer progression. Int J Biol Sci 2020, 16(6), 1023-34.

Tumor-associated macrophages (TAMs) are closely correlated with tumor occurrence, invasion, and metastasis. However, factors affecting the biological functions of TAMs in colorectal cancer (CRC) are incompletely understood. Here, we found that Wnt5a was mainly expressed on TAMs of tumor stroma but not on CRC cells. Subsequently, we found that Wnt5a+ TAMs facilitated tumor cell proliferation and migration, and recruited macrophages infiltration. Furthermore, Wnt5a knockdown impaired the pro-tumor roles of TAMs in vivo and in vitro. Mechanistically, the cancer-promoting roles of Wnt5a in TAMs depended on CaMKII-ERK pathway-mediated CCL2 secretion. Our data reveal the crucial role played by TAM-expressed Wnt5a in CRC tumorigenesis through paracrine secretion of CCL2. We first report the connection between Wnt5a/CaMKII/ERK/CCL2 axis and biological functions of TAMs in tumor microenvironment, indicating that Wnt5a may be a novel therapeutic target for CRC.

5 illus, 43 ref

043162   NIU H, HUANG Y, YAN L, ZHANG L, ZHAO M, LU T, YANG X, CHEN Z, ZHAN C, et al. (Thoracic Surgery Dep, Fudan Univ, Shanghai, China, Email: czhan10@fudan.edu.cn) : Knockdown of SMAD3 inhibits the growth and enhances the radiosensitivity of lung adenocarcinoma via p21 in vitro and in vivo. Int J Biol Sci 2020, 16(6), 1010-22.

Radiotherapy is an effective approach for the treatment of lung adenocarcinoma. However, evidence suggests that lung adenocarcinoma can easily develop tolerance to radiotherapy. The purpose of this study was to investigate the effect and mechanism of SMAD3 on the radiosensitivity of lung adenocarcinoma in vitro and in vivo. We found that knockdown of SMAD3 using two short hairpin RNAs in lentivirus vectors significantly inhibited cell growth and increased radiosensitivity of the lung adenocarcinoma cell lines A549, H1299, and H1975. Using RNA sequencing and bioinformatics analyses, we found that the significantly differentially expressed genes in SMAD3 knockdown cells were mainly enriched in the cell cycle process. We then showed that knockdown of SMAD3 significantly reduced expression of cyclin-dependent kinase inhibitor 1 (p21) and increased the proportion of G2/M phase cells and the radiosensitivity of lung adenocarcinoma. Chromatin immunoprecipitation results in the Gene Expression Omnibus (GEO) database and our luciferase assay verified that SMAD3 directly bound the p21 promoter. A series of rescue experiments showed that overexpression of p21 partly reversed the effect of SMAD3 on proliferation and radioresistance in vitro and in vivo. Moreover, we found that the expression levels of SMAD3 and p21 were highly correlated, and both correlated with the patients’ survival in online databases and clinical specimens. Expression of SMAD3 and p21 was also significantly different between radioresistant and radiosensitive patients in our hospital. Our results indicate that SMAD3 is a potential prognosis and radiosensitivity indicator as well as a target for radiotherapy and other treatments of patients with lung adenocarcinoma.

8 illus, 1 table, 33 ref

043161   SUN X, XIAO L, CHEN J, CHEN X, CHEN X, YAO S, LI H, ZHAO G, MA J (Orthopaedics Dep, Xi'an Jiaotong Univ, Xi'an 710054, Shaanxi, China, Email: zeltasun@163.com) : DNA methylation is involved in the pathogenesis of osteoarthritis by regulating CtBP expression and CtBP-mediated signaling. Int J Biol Sci 2020, 16(6), 994-1009.

Osteoarthritis (OA) is a common type of arthritis. Chronic inflammation is an important contributor to the pathogenesis of OA. The maturation and secretion of proinflammatory cytokines are controlled by inflammasomes, especially NLRP1 (NLR Family Pyrin Domain Containing 1) and NLRP3. In this study, we identified a transactivation mechanism of NLRP3 mediated by CtBPs (C-terminal-binding proteins). We found that both the mRNA and protein levels of CtBPs were significantly increased in OA biopsies. Analyzing the profiles of differentially expressed genes in CtBP-knockdown and overexpression cells, we found that the expression of NLRP3 was dependent on CtBP levels. By the knockdown or overexpression of transcription factors that potentially bind to the promoter of NLRP3, we found that only AP1 could specifically regulate the expression of NLRP3. Using immunoprecipitation (IP) and Co-IP assays, we found that AP1 formed a transcriptional complex with a histone acetyltransferase p300 and CtBPs. The knockdown of any member of this transcriptional complex resulted in a decrease in the expression of NLRP3. To explore the underlying mechanism of CtBP overexpression, we analyzed their promoters and found that they were abundant in CpG islands. Treatment with the DNA methylation inhibitor 5-aza-2′- deoxycytidine (AZA) or knockdown of DNMTs (DNA methyltransferases) resulted in the overexpression of CtBPs, while overexpression of DNMTs caused the reverse effects on CtBP expression. Collectively, our results suggest that the decreased DNA methylation levels in the promoters of CtBPs upregulate their expression. Increased CtBPs associated with p300 and AP1 to form a transcriptional complex and activate the expression of NLRP3 and its downstream signaling, eventually aggravating the inflammatory response and leading to the pathogenesis of OA.

8 illus, 51 ref

043160   JIANG Y, LIN L, CHEN S, JIANG L, KRIEGBAUM M C, GARDSVOLL H, HANSEN L V, LI J, PLOUG M, YUAN C, et al. (Minjiang Univ, Fuzhou, 350108, China, Email: HMD_lab@fzu.edu.cn) : Crystal structures of human C4.4A reveal the unique association of Ly6/uPAR/α-neurotoxin domain. Int J Biol Sci 2020, 16(6), 981-93.

Ly6/uPAR/α-neurotoxin domain (LU-domain) is characterized by the presence of 4–5 disulfide bonds and three flexible loops that extend from a core stacked by several conversed disulfide bonds (thus also named three-fingered protein domain). This highly structurally stable protein domain is typically a protein-binder at extracellular space. Most LU proteins contain only single LU-domain as represented by Ly6 proteins in immunology and α-neurotoxins in snake venom. For Ly6 proteins, many are expressed in specific cell lineages and in differentiation stages, and are used as markers. In this study, we report the crystal structures of the two LU-domains of human C4.4A alone and its complex with a Fab fragment of a monoclonal anti-C4.4A antibody. Interestingly, both structures showed that C4.4A forms a very compact globule with two LU-domain packed face to face. This is in contrast to the flexible nature of most LU-domain-containing proteins in mammals. The Fab combining site of C4.4A involves both LU-domains, and appears to be the binding site for AGR2, a reported ligand of C4.4A. This work reports the first structure that contain two LU-domains and provides insights on how LU-domains fold into a compact protein and interacts with ligands.

6 illus, 1 table, 88 ref

043159   LIAO L, ZHANG S, ZHAO L, CHANG X, HAN L, HUANG J, CHEN D (Orthopedic Surgery Dep, Rush Univ Medical Center, Chicago, Illinois, USA, Email: jian_huang@rush.edu) : Acute synovitis after trauma precedes and is associated with osteoarthritis onset and progression. Int J Biol Sci 2020, 16(6), 970-80.

Osteoarthritis (OA) is a whole-joint disease characterized by cartilage destruction, subchondral bone sclerosis, osteophyte formation, and synovitis. However, it remains unclear which part of the joint undergoes initial pathological changes that drives OA onset and progression. In the present study, we investigated the longitudinal alterations of the entire knee joint using a surgically-induced OA mouse model. Histology analysis showed that synovitis occurred as early as 1 week after destabilization of the medial meniscus (DMM), which preceded the events of cartilage degradation, subchondral sclerosis and osteophyte formation. Importantly, key pro-inflammatory cytokines such as IL-1β, IL-6, TNFα, and Ccl2, major matrix degrading enzymes including Adamts4, Mmp3 and Mmp13, as well as nerve growth factor (NGF), all increased significantly in both synovium and articular cartilage. It is notable that the inductions of these factors in synovium are far more extensive than those in articular cartilage. Results from behavioral tests demonstrated that sensitization of knee joint pain developed after 8 weeks, later than histological and molecular changes. In addition, the nanoindentation modulus of the medial tibiae decreased 4 weeks after DMM surgery, simultaneous with histological OA signs, which is also later than appearance of synovitis. Collectively, our data suggested that synovitis precedes and is associated with OA, and thus synovium may be an important target to intervene in OA treatment.

7 illus, 1 table, 38 ref

043158   ZHOU Y, ZHOU J, SUN B, XU W, ZHONG M, LI Y, HE C, CHEN Y, WANG X, JONES P M, et al. (Endocrinology Dep, Southeast Univ, Nanjing, China, Email: sunzilin1963@seu.edu.cn) : Vitamin A deficiency causes islet dysfunction by inducing islet stellate cell activation via cellular retinol binding protein 1. Int J Biol Sci 2020, 16(6), 947-56.

Vitamin A (VA) plays an essential role in pancreatic homeostasis. Islet stellate cells (ISCs) are VA-storing cells in pancreatic islets. Herein, we have investigated the effect of VA on glucose homeostasis trough regulation of ISCs function in dietary VA deficiency model mice. Male C57BL/6 mice were randomly fed a VA-sufficient, a VA-deficient (VAD) or a VAD-rescued diet. Glucose metabolism was assessed by glucose tolerance tests and immunohistochemistry. ISCs activation degree was evaluated by immunofluorescence, quantitative PCR and western blotting in both, retinol-treated cultured ISCs and model mice. Changes in ISCs phenotype and their effect on islets were assessed by lentiviral transduction and enzyme-linked immunosorbent assays in a co-culture system. VAD mice showed irregular shaped islet, glucose intolerance, islet size distribution excursions, and upregulated expression of α-smooth muscle actin (α-SMA, marker of ISCs activation). Reintroduction of dietary VA restored pancreatic VA levels, endocrine hormone profiles, and inhibited ISCs activation. Incubation with retinol increased the expression of VA signaling factors in ISCs, including cellular retinol binding protein 1 (CRBP1). The knockdown of CRBP1 maintained the quiescent ISCs phenotype and reduced the damage of activated ISCs on islet function. VA deficiency reduced islet function by activating ISCs in VAD mice. Restoring ISCs quiescence via CRBP1 inhibition could reverse the impairment of islet function caused by activated ISCs exposure.

4 illus, 44 ref

043157   LIU S-H, WANG P-P, CHEN C-T, LI D, LIU Q-Y, LV L, LIU X, WANG L-N, LI B-X, WENG C-Y, et al. (Oncology Dep, South China Univ of Technology, Guangzhou, Guangdong 510180, China, Email: eywuyong@scut.edu.cn) : MicroRNA-148b enhances the radiosensitivity of B-cell lymphoma cells by targeting Bcl-w to promote apoptosis. Int J Biol Sci 2020, 16(5), 935-46.

Lymphoma is a malignant disease of the hematopoietic system that typically affects B cells. The up-regulation of miR-148b is associated with radiosensitization in B-cell lymphoma (BCL). This study aimed to explore the role of miR-148b in regulating the radiosensitivity of BCL cells and to investigate the underlying mechanism. miR-148b directly targeted Bcl-w, decreased the cell viability and colony formation, while promoted apoptosis, in irradiated BCL cells. These changes were accompanied by decreased mitochondrial membrane potential, release of cytochrome C, increased levels of the cleaved caspase 9 and caspase 3, and increased expression of other proteins related to the mitochondrial apoptosis pathway. These effects of miR-148b were effectively inhibited by Bcl-w. In addition, miR-148b inhibited the growth of tumors in nude mice implanted with xenografts of irradiated Raji cells. In patients with BCL, levels of miR-148b were downregulated, while levels of Bcl-w were upregulated; a significant negative correlation between levels of miR-148b and Bcl-w was confirmed. Taken together, these experiments showed that miR-148b promoted radiation-induced apoptosis in BCL cells by targeting anti-apoptotic Bcl-w. miR-148b might be used as a marker to predict the radiosensitivity of BCL.

9 illus, 2 tables, 44 ref

043156   YE F, WANG Y, HE Q, CUI C, YU H, LU Y, ZHU S, XU H, ZHAO X, YIN H, et al. (Foshan Univ, Foshan, 528231, Guangdong, China, Email: zhuqingsicau@163.com) : Exosomes transmit viral genetic information and immune signals may cause immunosuppression and immune tolerance in ALV-J Infected HD11 cells. Int J Biol Sci 2020, 16(5), 904-20.

Avian leukosis virus (ALV) is oncogenic retrovirus that not only causes immunosuppression but also enhances the host’s susceptibility to secondary infection. Exosomes play vital role in the signal transduction cascades that occur in response to viral infection. We want to explore the function of exosomes in the spread of ALV and the body’s subsequent immunological response. RNA-sequencing and the isobaric tags for relative and absolute quantitation (iTRAQ) method were used to detect differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) in exosomes secreted by macrophage cells in response to injection with ALV subgroup J (ALV-J). RNA-sequencing identified 513 DEGs in infected cells, with specific differential regulation in mRNA involved in tight junction signaling, TNF signaling, salmonella infection response, and immune response, among other important cellular processes. Differential regulation was observed in 843 lncRNAs, with particular enrichment in those lncRNA targets involved in Rap1 signaling, HTLV-I infection, tight junction signaling, and other signaling pathways. A total of 50 DEPs were identified in the infected cells by iTRAQ. The proteins enriched are involved in immune response, antigen processing, the formation of both MHC protein and myosin complexes, and transport. Combined analysis of the transcriptome and proteome revealed that there were 337 correlations between RNA and protein enrichment, five of which were significant. Pathways that were enriched on both the RNA and protein levels were involved in pathways in cancer, PI3K-Akt signaling pathway, Endocytosis, Epstein-Barr virus infection. These data show that exosomes are transmitters of intercellular signaling in response to viral infection. Exosomes can carry both viral nucleic acids and proteins, making it possible for exosomes to be involved in the viral infection of other cells and the transmission of immune signals between cells. Our sequencing results confirme previous studies on exosomes and further find exosomes may cause immunosuppression and immune tolerance.

13 illus, 6 tables, 54 ref