043291   HUANG B, ZHOU Z, LIU J, WU X, LI X, HE Q, ZHANG P, TANG X (Guangdong Medical Univ, Zhanjiang, Guangdong 524023, P.R. China, Email: tangxudong2599@126.com) : The role of monoamine oxidase A in HPV-16 E7-induced epithelial-mesenchymal transition and HIF-1α protein accumulation in non-small cell lung cancer cells. Int J Biol Sci 2020, 16(14), 2692-703.

Our previous studies have found that human papillomavirus (HPV)-16 E7 oncoprotein promotes epithelial-mesenchymal transition (EMT) and hypoxia-inducible factor-1α (HIF-1α) protein accumulation in non-small cell lung cancer (NSCLC) cells and monoamine oxidase A (MAOA) is highly expressed in NSCLC tissues. Here, we further explored the role of MAOA in HPV-16 E7-induced EMT and HIF-1α protein accumulation in A549 and NCI-H460 NSCLC cells. Our results showed that HPV-16 E7 enhanced MAOA expression in NSCLC cells. Additionally, MAOA knockout inhibited HPV-16 E7-induced migration, invasion, and EMT, and significantly reduced HPV-16 E7-induced ROS generation and HIF-1α protein accumulation via promoting its degradation. Furthermore, MAOA knockout suppressed HPV-16 E7-induced ERK1/2 activation. In vivo, MAOA knockout inhibited tumor growth, metastasis, and the expression of EMT-related markers and HIF-1α proteins induced by HPV-16 E7 in NCI-H460 NSCLC subcutaneous xenograft and in situ intrapulmonary models of nude mice. Taken together, our findings provide evidence that MAOA plays a key role in EMT and HIF-1α protein accumulation induced by HPV-16 E7 in NSCLC cells, suggesting that MAOA may be a potential therapeutic target for HPV-related NSCLC.

7 illus, 53 ref

041485   ACHARYA R, PATTANAYAK S P ( Pharmaceutical Sciences and Technology Dep, Birla Institute of Technology, Ranchi - 835 215, Jharkhand, Email: sppattanayak@bitmesra.ac.in) : Small molecules polypharmacology for the treatment of breast cancer: A roadmap for the future chemotherapy. Indian J Pharm Edu Res 2020, 54(2), 225-33.

Breast cancer is considered as the most common ailment in women across the globe. There are a variety of major factors for the growth and progression of breast cancer. Although genetic changes are key factors for cancer progression, epigenetic and environmental factors also have their fair share of contribution. An attempt has been made in the review to highlight about small molecule polypharmacology for the treatment of breast cancer which is the design of small molecules (molecular weight <900Da) that aim for two or more cancer targets (tyrosine kinases). Small molecules are advantageous over conventional chemotherapy with higher target specificity, lesser known side effects, simplified dosing schedule, non-invasive dosing and improved patient compliance. Small molecules have been introduced since the past two decades and has been a potential success in the field of cancer chemotherapy, but there are lot more to explore. The review mainly focuses on the essential cellular signaling pathways involved in breast cancer progression and the recent small molecule inhibitors that show promising activity against the pathways.

4 illus, 1 table, 65 ref

043290   ZHOU Y, JIA E, PAN M, ZHAO X, GE Q (Southeast Univ, Nanjing 210096, China, Email: geqinyu@seu.edu.cn) : Encoding method of single-cell spatial transcriptomics sequencing. Int J Biol Sci 2020, 16(14), 2663-74.

Despite significant advances in parallel single-cell RNA sequencing revealing astonishing cellular heterogeneity in many tissue types, the spatial information in the tissue context remains missing. Spatial transcriptome sequencing technology is designed to distinguish the gene expression of individual cells in their original location. The technology is important for the identification of tissue function, tracking developmental processes, and pathological and molecular detection. Encoding the position information is the key to spatial transcriptomics because different methods have different encoding efficiencies and application scenarios. In this review, we focus on the latest technologies of single-cell spatial transcriptomics, including technologies based on microwell plates, barcoded bead arrays, microdissection, in situ hybridization, and barcode in situ targeting, as well as mixed separation-based technologies. Moreover, we compare these encoding methods for use as a reference when choosing the appropriate technology.

3 illus, 1 table, 68 ref

043289   CHEN Y-T, HUANG C-R, CHANG C-L, CHIANG J-Y, LUO C-W, CHEN H-W, YIP H-K (Surgery Dep, Medicine Univ Coll, Kaohsiung City 83301, Taiwan) : Jagged2 progressively increased expression from stage i to iii of bladder cancer and melatonin-mediated downregulation of notch/jagged2 suppresses the bladder tumorigenesis via inhibiting PI3K/AKT/mTOR/MMPs signaling. Int J Biol Sci 2020, 16(14), 2648-62.

This study assessed the expression of Jagged2 in human bladder cancer (BC) tested the hypothesis that melatonin (Mel) inhibited the tumorigenesis of BC cells mainly through downregulating the Notch/Jagged2 and PI3K/AKT/mTOR/MMPs^(2&9) signaling pathways. Tissue array from BC patients showed that the gene and protein expressions of JAG2/Jagged2 were significantly upregulated from T1 to T3 (primary tumor size) and from stage I to III (all p<0.001). In vitro study showed that in BC cell line of UMUC3, the cellular and protein expressions of Jagged2 were significantly attenuated in Mel-treated UMUC3 and further attenuated in UMUC3 shRNA silenced Notch/JAG2 (UMUC3^KD) than in UMUC3 only (all p<0.0001). The protein expressions of Notch/Jagged2/MMPs^(2&9)/PI3K/p-AKT/mTOR/p53/ratio of LC3BII/LC3B-I were significantly progressively reduced from UMUC3 to UMUC3+Mel/1.0mM, further to UMUC3+Mel/2.0mM and furthermore to UMUC3^KD (all p<0.0001). The cell proliferation/invasion/colony formation/healing-process were significantly inhibited in Mel-treated/2.0mM UMUC3 and further significantly inhibited in UMUC3^KD regardless of Mel treatment as compared with UMUC3 only (all p<0.0001). By day 28 after UMUC3 implanted into nude mouse back, the BC weight/volume were significantly reduced in UMUC3+Mel (100 mg/kg/day) and furthermore reduced in UMUC3^KD (all p<0.0001) as compared with UMUC3 only (all p<0.0001). The cellular (MMPs^(2&9)/Notch/Jagged2) and protein (Notch/Jagged2/PI3K/p-AKT/mTOR/MMPs^(2&9)) exhibited a similar trend, whereas the PTEN protein level exhibited an opposite pattern of PI3K among three groups (all p<0.0001). Notch/Jagged-PI3K/p-AKT/mTOR/MMPs is one essential signaling pathway for BC survival, proliferation and invasion that were remarkably suppressed by Mel treatment.

12 illus, 41 ref

041482   JAGWANI S, JALALPURE S, DHAMECHA D, HUA G S, JADHAV K (KLE Coll of Pharmacy, Belagavi - 590 010 Karnataka, Email: jalalpuresunil@rediffmail.com) : Development and validation of reverse-phase high-performance liquid chromatographic method for determination of resveratrol in human and rat plasma for preclinical and clinical studies. Indian J Pharm Edu Res 2020, 54(1), 187-93.

The aim of the study was to develop and validate a simple and precise reverse-phase High Performance Liquid Chromatography (RP-HPLC) method for quantitative analysis of trans-resveratrol in human and rat plasma. HPLC method was developed by using Phenomenex Luna C₁₈ column (150 x 4.6 mm, 5 µm) and the optimized mobile phase comprised of acetonitrile/water in isocratic mode (30:70, v/v) with the flow rate of 1.0 mL/min. Trans-resveratrol was detected at a UV wavelength of 306 nm. Developed method was validated as per International Conference on Harmonization (ICH) M10 guidelines. The proposed method was found simple, precise and linear with regression coefficient of 0.999 which could analyse the samples in nanograms levels with mean percent recovery in the acceptable range of 94.44 – 97.44 %. The method was precise at the intra-day and inter-day levels as reflected by the relative standard deviation values (less than 3.36 %). Trans resveratrol was found to be stable in plasma under different storage conditions. The present investigation demonstrated that the developed method was succesfully applied to accurately determine transresveratrol in human and rat plasma and therefore can be applicable for pre-clinical and clinical studies.

3 illus, 6 tables, 16 ref

041481   NAGARAJAPPA M H, SRIVATSA H S (M. S. Ramaiah Univ of Applied Sciences, Bengaluru - 560 054, Karnataka, Email: hs.srivatsa@gmail.com) : Modelling customers’ buying behaviour of Jan Aushadhi (Generic Medicines). Indian J Pharm Edu Res 2020, 54(1), 175-86.

Jan Aushadhi, a scheme to make affordable generic drugs available to large sections of population was launched by the Government of India across the country in the year 2008. This exploratory research was conducted to study the attitude of customers (Bengaluru, India) towards acceptance of Jan Aushadhi and an attempt was made to model buying behavior in order to suggest mechanisms to speed up the acceptance. Data was collected using structured questionnaire. Z-test and Exploratory factor analysis was conducted for hypothesis testing. Discriminant analysis was conducted to model consumption. The variables that discriminated consumers from non-consumers were ‘effectiveness’, ‘doctor’s opinion’, ‘lower price’, ‘Quality’, ‘less expensive than other branded medicines’, ‘doctor’s prescription’, ‘convenience’, ‘doctor informs’, ‘home delivery’ and ‘brand reputation’. Results found that ‘doctor’s prescription’, ‘lower price’, ‘availability of Jan Aushadhi outlet’, ‘quality of generic medicine’ and ‘recommendation from others’ had a significant influence on acceptance of Jan Aushadhi. Therefore, the study recommends the doctors to prescribe generic medicine, increase the number of Jan Aushadhi outlets and provide awareness about the quality and efficacy of Jan Aushadhi.

13 tables, 23 ref

041480   CHOHAN M S, ELGORASHE R E E, BALGONAME A A, ATTIMARAD M, SREEHARSHA N, VENUGOPALA K N, NAIR A B, POTTATHIL S (Pharmaceutical Sciences Dep, King Faisal Univ, Al-Ahsa-31982, Saudi Arabia, Email: mattimarad@kfu.edu.sa) : Eco-friendly derivative UV spectrophotometric methods for simultaneous determination of diclofenac sodium and moxifloxacin in laboratory mixed ophthalmic preparation. Indian J Pharm Edu Res 2020, 54(1), 166-74.

Diclofenac Sodium (DCL) and Moxifloxacin HCl (MOX) were simultaneously used after cataract surgery to reduce the post-operative inflammation and to control infection respectively. Three simple, accurate, eco-friendly and reproducible UV spectroscopic methods were established for concurrent determination of diclofenac sodium and moxifloxacin in ophthalmic preparation without prior separation. The first technique was established on the measurement of a peak amplitude of the first derivative spectra at the zero-crossing wavelength of one analyte. The second method was the determination of peak amplitude difference between peak and trough of ratio spectra. The third method involves the measurement of the peak amplitude of the first derivative of ratio spectra. Water has been used as a solvent. The analytes exhibited good linearity in the range of 1 -15 µg/mL for DCL and 1-18 µg/mL for MOX with excellent correlation coefficient (r²>0.999). Low percent relative standard deviation confirmed the precision of the methods. Excellent recovery with low percent relative error proved the accuracy of the methods. The specificity of the methods was evaluated by analyzing the laboratory prepared solutions of DCL and MOX. Proposed three techniques were effectively utilized for the simultaneous determination of DCL and MOX from ophthalmic preparation. The outcomes of the proposed procedures were compared with the earlier described methods and no statistical difference was found between the methods in terms of accuracy and precision.

7 illus, 5 tables, 29 ref

041479   PATEL K G, PATEL S G K, SHAH P A, TANDEL D B, GANDHI T R (Quality Assurance Dep, Anand Pharmacy Coll, Anand, Gujarat, Email: kalpana_jpatel@ yahoo.com) : Development and validation of HPTLC Method along with forced degradation study for the simultaneous estimation of azelastine hydrochloride and fluticasone propionate in nasal spray formulation using design of experiment approach. Indian J Pharm Edu Res 2020, 54(1), 155-65.

The present research study focuses on the development of the highperformance thin layer chromatographic method using design of experiment approach for the simultaneous estimation of Azelastine hydrochloride and Fluticasone propionate along with forced degradation study. High performance thin layer chromatographic separation was performed on aluminium plates precoated with silica gel 60 F₂₅₄ using toluene: chloroform: methanol (5: 4: 2, v/v/v) as optimized mobile phase. Azelastine hydrochloride and Fluticasone propionate were exposed to different forced degradation conditions. Full factorial design was applied on acid and base induced degradation and statistical analysis by ANOVA was performed with interpretation of various plots. The method was validated by determination of linearity, precision, accuracy, specificity and robustness according to ICH guidelines. From different forced degradation conditions, major degradation was observed in acidic and basic condition. R_f for Azelastine hydrochloride and Fluticasone propionate was 0.33 and 0.53 at 232 nm. The factor, temperature showed maximum % contribution in acid and base induced degradation. Linear concentration range was 280-1680 ng/band for Azelastine hydrochloride, 100-600 ng/band for Fluticasone propionate. The % recovery ranged within 101.15- 102.65 % for Azelastine hydrochloride and 99.09-103.40 % for Fluticasone propionate. The % R.S.D values were less than 2 % for both drugs indicating that the method is accurate, sensitive and precise. In summary, a novel, simple, accurate and reproducible high-performance thin layer chromatographic method was developed, for routine quality control testing of pharmaceutical formulation.

16 illus, 5 tables, 29 ref

043288   ZHANG J, GAO S, ZHANG Y, YI H, XU M, XU J, LIU H, DING Z, HE H, WANG H, et al. (Jilin Univ First Hospital of Changchun, China, Email: wei_feng@jlu.edu.cn) : MiR-216a-5p inhibits tumorigenesis in pancreatic cancer by targeting TPT1/mTORC1 and is mediated by LINC01133. Int J Biol Sci 2020, 16(14), 2612-27.

MiR-216a-5p has opposite effects on tumorigenesis and progression in the context of different tumors, acting as either a tumor suppressor or an oncogene. However, the expression and function of miR-216a-5p in pancreatic cancer (PC) is not well characterized. In this study, we found miR-216a-5p was significantly downregulated in PC tissues and cell lines, which showed a negative correlation with peripancreatic lymph, perineural invasion and TNM stage of PCs patients. We made use of functional assays to reveal that miR-216a-5p inhibited growth and migration of PC cells in vitro and in vivo. Then, by employing the bioinformatics analysis and luciferase reporter assay, we demonstrated TPT1 was a potential target of miR-216a-5p, which contributes to tumor malignance by mediating mTORC1 pathway-associated autophagy. Furthermore, bioinformatics analysis and RNA pulldown confirmed that miR-216a-5p was mediated by LINC01133, which sponge miR-216a-5p, as a competing endogenous RNA (ceRNA). Collectively, our study revealed an important role of LINC01133/miR-216a-5p/TPT1 axis in the genesis and progression of PCs, which provides potential biomarkers for clinical diagnosis and therapy of PCs.

7 illus, 1 table, 30 ref

043287   DING N, YOU A, TIAN W, GU L, DENG D (Peking Univ, Beijing, 100142, China, Email: dengdajun@bjmu.edu.cn) : Chidamide increases the sensitivity of non-small cell lung cancer to crizotinib by decreasing c-met mrna methylation. Int J Biol Sci 2020, 16(14), 2595-611.

Crizotinib is a kinase inhibitor targeting c-MET/ALK/ROS1 used as the first-line chemical for the treatment of non-small cell lung cancer (NSCLC) with ALK mutations. Although c-MET is frequently overexpressed in 35-72 % of NSCLC, most NSCLCs are primarily resistant to crizotinib treatment. A set of NSCLC cell lines were used to test the effect of chidamide on the primary crizotinib resistance in vitro and in vivo. Relationships between the synergistic effect of chidamide and c-MET expression and RNA methylation were systemically studied with a battery of molecular biological assays. We found for the first time that chidamide could sensitize the effect of crizotinib in a set of ALK mutation-free NSCLC cell lines, especially those with high levels of c-MET expression. Notably, chidamide could not increase the sensitivity of NSCLC cells to crizotinib cultured in serum-free medium without hepatocyte growth factor (HGF; a c-MET ligand). In contrast, the addition of HGF into the serum-/HGF-free medium could restore the synergistic effect of chidamide. Moreover, the synergistic effect of chidamide could also be abolished either by treatment with c-MET antibody or siRNA-knockdown of c-MET expression. While cells with low or no c-MET expression were primarily resistant to chidamide-crizotinib cotreatment, enforced c-MET overexpression could increase the sensitivity of these cells to chidamide-crizotinib cotreatment. Furthermore, chidamide could decrease c-MET expression by inhibiting mRNA N6-methyladenosine (m6A) modification through the downregulation of METTL3 and WTAP expression. Chidamide-crizotinib cotreatment significantly suppressed the activity of c-MET downstream molecules. Chidamide downregulated c-MET expression by decreasing its mRNA m6A methylation, subsequently increasing the crizotinib sensitivity of NSCLC cells in a c-MET-/HGF-dependent manner.

8 illus, 1 table, 47 ref

043286   LUO A, XU Y, LI S, BAO J, LU J, DING N, ZHAO Q, FU Y, LIU F, CHO W C, et al. (Tongji Univ School of Medicine, Shanghai 200120, China, Email: zuoren.yu@tongji.edu.cn) : Cancer stem cell property and gene signature in bonemetastatic breast cancer cells. Int J Biol Sci 2020, 16(14), 2580-94.

The majority of the deaths from breast cancer is due to metastasis. Bone is the most common organ to which breast cancer cells metastasize. The mechanism regulating the bone-metastatic preference remains unclear; there is a lack of a gene signature to distinguish bone-metastatic breast cancer cells. Herein, florescence-labeled MDA-MB-231 cells were transplanted into the fat pads of of the mammary gland in nude mice to generate breast tumors. Tumor cells invaded into the circulation were tracked by in vivo flow cytometry system. Metastatic tumor cells in the bone were isolated using fluorescent-activated cell sorting technique, followed by assays of cell colony formation, migration and invasion, mammosphere formation in vitro, mammary gland tumorigenesis in vivo, and Next-Generation Sequencing analysis as well. Through tumor regeneration and cell sorting, two bone-metastatic cell sublines were derived from MDA-MB-231 cells; which showed higher abilities to proliferate, migrate, invade and epithelial-to-mesenchymal transit in vitro, and stronger ability to regenerate tumors and metastasize to the bone in vivo. Both cell sublines exhibited cancer stem cell-like characteristics including higher expression levels of stem cell markers and stronger ability for mommaspheres formation. Furthermore, a Normal Distribution-like pattern of the bone-metastatic cells invading into circulation was firstly identified. Deep-sequencing analysis indicated upregulation of multiple signaling pathways in regulating EMT, cell membrane budding and morphologic change, lipid metabolism, and protein translation, which are required to provide adequate metabolic enzymes, structural proteins, and energy for the cells undergoing metastasis. In conclusion, we established two bone-metastatic breast cancer cell sublines, carrying higher degree of stemness and malignancy. The gene signature distinguishing the bone-metastatic breast cancer cells holds therapeutic potentials in prevention of breast cancer metastasis to the bone.

7 illus, 28 ref

043285   HUANG K-Y, QUE J-Q, HU Z-S, YU Y-W, ZHOU Y-Y, WANG L, XUE Y-J, JI K-T, ZHANG X-M (Cardiology Dep, Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical Univ, Wenzhou 325027, Zhejiang, China, Email: jikt@wzmc.edu.cn) : Metformin suppresses inflammation and apoptosis of myocardiocytes by inhibiting autophagy in a model of ischemia-reperfusion injury. Int J Biol Sci 2020, 16(14), 2559-79.

Metformin (Met) is a major widely used oral glucose lowering drug for the treatment of type 2 diabetes. It is reported that metformin could regulate autophagy in various diseases of cardiovascular system including in I/R injury, diabetic cardiomyopathy and heart failure. Autophagy plays a controversial role in ischemia/reperfusion (I/R) injury, and this research was performed to explore the cardioprotective effect of Met on I/R injury and discuss the underlying mechanism of autophagy in it. In vivo and in vitro, Met exerted cardioprotection function of decreasing myocardial inflammation and apoptosis with a decrease in the level of autophagy. Moreover, Met significantly inhibited autophagosome formation and restore the impairment of autophagosome processing, which lead to cardioprotection effect of Met. Akt was up-regulated in Met-treated I/R hearts and miransertib, a pan-AKT inhibitor, was able to reverse the alleviating autophagy effect of Met. We demonstrate that Met protects cardiomyocytes from I/R-induced apoptosis and inflammation through down regulation of autophagy mediated by Akt signaling pathway.

6 illus, 51 ref

043284   JIA C, WANG G, WANG T, FU B, ZHANG Y, HUANG L, DENG Y, CHEN G, WU X, CHEN J, et al. (Third Affiliated Hospital of Sun Yat-sen Univ, Guangzhou, China, Email: chgh1955@263.net) : Cancer-associated fibroblasts induce epithelialmesenchymal transition via the transglutaminase 2-dependent IL-6/IL6R/STAT3 axis in hepatocellular carcinoma. Int J Biol Sci 2020, 16(14), 2542- 58.

Cancer-associated fibroblasts (CAFs) play crucial roles in enhancing cell survival, proliferation, invasion, and metastasis. We previously showed that hepatocellular carcinoma-derived CAFs (H-CAFs) promoted proliferation of hepatocellular carcinoma (HCC) cells. This study aimed to further explore the role of CAFs in HCC epithelial-mesenchymal transition (EMT) and the underlying mechanism. High CAF density was significantly associated with liver cirrhosis, inferior clinicopathologic characteristics, elevated EMT-associated markers, and poorer survival in human HCC. Within HCC cells, EMT was induced after co-culture with H-CAFs. Secretomic analysis showed that IL-6 and HGF were the key EMT-stimulating cytokines secreted by H-CAFs. Proteomic analysis revealed that TG2 was significantly upregulated in HCC cells with EMT phenotypes. Overexpression of TG2 promoted EMT of HCC cells, and knockdown of TG2 remarkably attenuated the H-CAF-induced EMT. Furthermore, during EMT, TG2 expression was enhanced after HCC cells were stimulated by IL-6, but not HGF. Inhibition of the IL-6/STAT3 signaling decreased TG2 expression. The principal TG2 transcription control element and a potential STAT3 binding site were identified using promoter analysis. Hence, H-CAFs facilitates HCC cells EMT mediated by IL-6, which in turn activates IL-6/IL6R/STAT3 axis to promote TG2 expression.

8 illus, 3 tables, 41 ref

043283   MENG Z, YU Y, ZHANG Y, YANG X, LV X, GUANG F, HATCH G M, ZHANG M, CHEN L (Pharmacology Dep, Jilin Univ, Changchun, Jilin, China, Email: chenl@jlu.edu.cn) : Highly bioavailable berberine formulation improves glucocorticoid receptor-mediated insulin resistance via reduction in association of the glucocorticoid receptor with phosphatidylinositol-3-kinase. Int J Biol Sci 2020, 16(14), 2527-41.

Excess glucocorticoid (GC) production is known to induce obesity and insulin resistance through increased activation of the glucocorticoid receptor (GR). The molecular mechanism for the non-genomic effects of excessive circulating GC on the insulin-signalling pathway in skeletal muscle is unknown. The plant alkaloid berberine has been shown to attenuate insulin resistance and inhibit gluconeogenesis in type 2 diabetic animals. A highly bioavailable berberine formulation termed Huang-Gui solid dispersion (HGSD), is a preparation of berberine coupled to sodium caprate and this markedly improving berberines bioavailability. Here we examined how HGSD treatment attenuated GR-mediated alteration in PI3K signalling and insulin resistance in diabetic rats, dexamethasone-treated mice and in insulin resistant C2C12 skeletal muscle cells. Blood glucose and skeletal muscle GC levels were increased and insulin signalling impaired in skeletal muscle of type 2 diabetic rats compared to controls. Treatment of these animals with HGSD restored blood glucose and skeletal muscle GC levels to that of controls. Insulin resistant C2C12 skeletal muscle cells exhibited impaired insulin signalling compared to controls and treatment of HGSD and RU486, an antagonist of GR, restored insulin signalling to that of control cells. Administration of dexamethasone to mice increased GR/GRα-associated PI3K and reduced IRS1-associated PI3K, phosphorylated-AKT, and membrane GLUT4 translocation resulting in a higher blood glucose concentration compared to controls. HGSD treatment of these mice improved insulin resistance by reducing the association of GR/GRα with PI3K. Excess GC-induced insulin resistance is mediated by increased association of GR with PI3K and treatment with HGSD attenuates these effects. We hypothesize that HGSD may be a promising candidate drug for the treatment of type 2 diabetes by reducing the association of GR with PI3K in skeletal muscle.

7 illus, 41 ref

043282   SHAHID M, KIM M, JIN P, ZHOU B, WANG Y, YANG W, YOU S, KIM J (Surgery and Biomedical Sciences Dep, Cedars-Sinai Medical Center, Los Angeles, CA 90048, Email: Jayoung.kim@csmc.edu) : S-palmitoylation as a functional regulator of proteins associated with cisplatin resistance in bladder cancer. Int J Biol Sci 2020, 16(14), 2490-505.

Protein S-palmitoylation is a powerful post-translational modification that regulates protein trafficking, localization, turnover, and signal transduction. Palmitoylation controls several important cellular processes, and, if dysregulated, can lead to cancer, cardiovascular disease, and neurological disorders. The role of protein palmitoylation in mediating resistance to systemic cisplatin-based chemotherapies in cancer is currently unknown. This is of particular interest because cisplatin is currently the gold standard of treatment for bladder cancer (BC), and there are no feasible options after resistance is acquired. Using unbiased global proteomic profiling of purified S-palmitoylated peptides combined with intensive bioinformatics analyses, we identified 506 candidate palmitoylated proteins significantly enriched in cisplatin-resistant BC cells. One of these proteins included PD-L1, which is highly palmitoylated in resistant cells. Pharmacological inhibition of fatty acid synthase (FASN) suppressed PD-L1 palmitoylation and expression, which suggests the potential use of FASN-PD-L1-targeted therapeutic strategies in BC patients. Taken together, these results highlight the role of protein palmitoylation in mediating BC chemoresistance.

7 illus, 1 table, 71 ref

043281   CHEN X, YUAN W, LI Y, LUO J, HOU N (Guangzhou Medical Univ, Guangzhou 511436, China, Email: houning@gzhmu.edu.cn) : Role of Hippo-YAP1/TAZ pathway and its crosstalk in cardiac biology. Int J Biol Sci 2020, 16(13), 2454-63.

The Hippo pathway undertakes a pivotal role in organ size control and the process of physiology and pathology in tissue. Its downstream effectors YAP1 and TAZ receive upstream stimuli and exert transcription activity to produce biological output. Studies have demonstrated that the Hippo pathway contributes to maintenance of cardiac homeostasis and occurrence of cardiac disease. And these cardiac biological events are affected by crosstalk among Hippo-YAP1/TAZ, Wnt/β-catenin, Bone morphogenetic protein (BMP) and G-protein-coupled receptor (GPCR) signaling, which provides new insights into the Hippo pathway in heart. This review delineates the interaction among Hippo, Wnt, BMP and GPCR pathways and discusses the effects of these pathways in cardiac biology.

4 illus, 95 ref

043280   ZHENG X, PENG Q, WANG L, ZHANG X, HUANG L, WANG J, QIN Z (Guangxi Birth Defects Research and Prevention Institute, Nanning, Guangxi, 530003, China, Email: jiaicq@csu.edu.cn) : Serine/arginine-rich splicing factors: The bridge linking alternative splicing and cancer. Int J Biol Sci 2020, 16(13), 2442-53.

The serine/arginine-rich splicing factors (SRs) belong to the serine arginine-rich protein family, which plays an extremely important role in the splicing process of precursor RNA. The SRs recognize the splicing elements on precursor RNA, then recruit and assemble spliceosome to promote or inhibit the occurrence of splicing events. In tumors, aberrant expression of SRs causes abnormal splicing of RNA, contributing to proliferation, migration and apoptosis resistance of tumor cells. Here, we reviewed the vital role of SRs in various tumors and discussed the promise of analyzing mRNA alternative splicing events in tumor. Further, we highlight the challenges and discussed the perspectives for the identification of new potential targets for cancer therapy via SRs family members.

5 illus, 1 table, 138 ref

043279   YANG D, LIU H-Q, LIU F-Y, TANG N, GUO Z, MA S-Q, AN P, WANG M-Y, WU H-M, YANG Z, et al. (Cardiology Dep, Renmin Hospital of Wuhan Univ, Wuhan 430060, RP China, Email: qztang@whu.edu.cn) : The roles of noncardiomyocytes in cardiac remodeling. Int J Biol Sci 2020, 16(13), 2414-29.

Cardiac remodeling is a common characteristic of almost all forms of heart disease, including cardiac infarction, valvular diseases, hypertension, arrhythmia, dilated cardiomyopathy and other conditions. It is not merely a simple outcome induced by an increase in the workload of cardiomyocytes (CMs). The remodeling process is accompanied by abnormalities of cardiac structure as well as disturbance of cardiac function, and emerging evidence suggests that a wide range of cells in the heart participate in the initiation and development of cardiac remodeling. Other than CMs, there are numerous noncardiomyocytes (non-CMs) that regulate the process of cardiac remodeling, such as cardiac fibroblasts and immune cells (including macrophages, lymphocytes, neutrophils, and mast cells). In this review, we summarize recent knowledge regarding the definition and significant effects of various non-CMs in the pathogenesis of cardiac remodeling, with a particular emphasis on the involved signaling mechanisms. In addition, we discuss the properties of non-CMs, which serve as targets of many cardiovascular drugs that reduce adverse cardiac remodeling.

3 illus, 1 table, 149 ref

043278   WU Y, MIN J, GE C, SHU J, TIAN D, YUAN Y, ZHOU D (First Affiliated Hospital of Anhui Medical Univ, Hefei, Anhui, 230022, China, Email: ayfy_zhoudian@163.com) : Interleukin 22 in liver injury, inflammation and cancer. Int J Biol Sci 2020, 16(13), 2405-13.

Interleukin 22(IL-22), a member of the IL-10 cytokine family and is an emerging CD4+Th cytokine that plays an important role in anti-microbial defense, homeostasis and tissue repair. We are interested in IL-22 as it has the double function of suppressing or encouraging inflammation in various disease models including hepatic inflammation. As a survival factor for hepatocytes, IL-22 plays a protective role in many kinds of liver diseases, such as hepatitis, liverfibrosis, or hepatocellular carcinoma (HCC) by binding to the receptors IL-22R1 and IL-10R2. Overexpression of IL-22 reduces liver fibrosis by attenuating the activation of hepatic stellate cell (the main cell types involved in hepatic fibrosis), and down-regulating the levels of inflammatory cytokines. Administration of exogenous IL-22 increases the replication of hepatocytes by inhibiting cell apoptosis and promoting mitosis, ultimately plays a contributing role in liver regeneration. Furthermore, treatment with IL-22 activates hepatic signal transducer and activator of transcription 3 (STAT3), ameliorates hepatic oxidative stress and alcoholic fatty liver, effectively alleviate the liver damage caused by alcohol and toxicant. In conclusion, the hepatoprotective functions and liver regeneration promoting effect of IL-22 suggests the therapeutic potential of IL-22 in the treatment of human hepatic diseases.

2 illus, 87 ref

043277   CHEN J, DENG X, LIU Y, TAN Q, HUANG G, CHE Q, GUO J, SU Z (Guangdong Pharmaceutical Univ, Guangzhou 510006, China, Email: suzhq@scnu.edu.cn) : Kupffer cells in non-alcoholic fatty liver disease: Friend or foe?. Int J Biol Sci 2020, 16(13), 2367-78.

The prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing all around the world and it may become the primary cause of terminal liver disease in adults and children in the next few decades. However, the pathogenesis of NAFLD is complex, and the Food and Drug Administration (FDA) has not approved any drugs for its treatment. Kupffer cells are the key cells regulating immunity in the liver, and the effect of their unique polarization on NAFLD has received increasing attention. Kupffer cells mainly reside in the lumen of hepatic sinusoids and account for 80 % to 90 % of colonized macrophages in the human body. They are phagocytic cells with the capacity for self-renewal that rarely migrate from their niche in the liver, and play a crucial role in regulating and maintaining homeostasis. Upon liver damage, Kupffer cells will be activated, releasing a good deal of inflammatory cytokines and chemokines. This review summarizes the multiple roles of Kupffer cells in the pathogenesis of NAFLD, the role of infiltrating macrophages in the pathogenesis of NAFLD is also briefly discussed, and aims to provide a theoretical basis for designing an NAFLD treatment strategy with Kupffer cells as the therapeutic target.

3 illus, 2 tables, 118 ref

043276   YANG X, LU D, ZHUO J, LIN Z, YANG M, XU X (Surgery Dep, Zhejiang Univ School of Medicine, Hangzhou, 310003, China, Email: zjxu@zju.edu.cn) : The gut-liver axis in immune remodeling: New insight into liver diseases. Int J Biol Sci 2020, 16(13), 2357-66.

The gut microbiota consists of a dynamic multispecies community of bacteria, fungi, archaea, and protozoans, playing a fundamental role in the induction, training, and function of the host immune system. The liver is anatomically and physiologically linked to the gut microbiota via enterohepatic circulation, specifically receiving intestine-derived blood through the portal vein. The gut microbiota is crucial for maintaining immune homeostasis of the gut-liver axis. A shift in gut microbiota composition can result in activation of the mucosal immune response causing homeostasis imbalance. This imbalance results in translocation of bacteria and migration of immune cells to the liver, which is related to inflammation-mediated liver injury and tumor progression. In this review, we outline the role of the gut microbiota in modulating host immunity and summarize novel findings and recent advances in immune-based therapeutics associated with the gut–liver axis. Moving forward, a deep understanding of the microbiome-immune-liver axis will provide insight into the basic mechanisms of gut microbiota dysbiosis affecting liver diseases.

1 illus, 1 table, 119 ref

043275   LI J, HUANG L, ZHAO H, YAN Y, LU J (Pathophysiology Dep, Zhengzhou Univ, Zhengzhou Univ, Email: lujing@zzu.edu.cn) : The role of interleukins in colorectal cancer. Int J Biol Sci 2020, 16(13), 2323-39.

Despite great progress has been made in treatment strategies, colorectal cancer (CRC) remains the predominant life-threatening malignancy with the feature of high morbidity and mortality. It has been widely acknowledged that the dysfunction of immune system, including aberrantly expressed cytokines, is strongly correlated with the pathogenesis and progression of colorectal cancer. As one of the most well-known cytokines that were discovered centuries ago, interleukins are now uncovering new insights into colorectal cancer therapy. Herein, we divide currently known interleukins into 6 families, including IL-1 family, IL-2 family, IL-6 family, IL-8 family, IL-10 family and IL-17 family. In addition, we comprehensively reviewed the oncogenic or antitumour function of each interleukin involved in CRC pathogenesis and progression by elucidating the underlying mechanisms. Furthermore, by providing interleukins-associated clinical trials, we have further driven the profound prospect of interleukins in the treatment of colorectal cancer.

4 illus, 2 tables, 143 ref

043274   KUMARI A, PATHAK D P, ASTHANA S (Translational Health Science and Technology Institute, Gurgaon Expressway-121 001, Haryana, Email: sasthana@thsti.res.in) : Bile acids mediated potential functional interaction between FXR and FATP5 in the regulation of lipid metabolism. Int J Biol Sci 2020, 16(13), 2308-22.

Perturbation in lipid homeostasis is one of the major bottlenecks in metabolic diseases, especially Non-alcoholic Fatty Liver Disease (NAFLD), which has emerged as a leading global cause of chronic liver disease. The bile acids (BAs) and their derivatives exert a variety of metabolic effects through complex and intertwined pathways, thus becoming the attractive target for metabolic syndrome treatment. To modulate the lipid homeostasis, the role of BAs, turn out to be paramount as it is essential for the absorption, transport of dietary lipids, regulation of metabolic enzymes and transporters that are essential for lipid modulation, flux, and excretion. The synthesis and transport of BAs (conjugated and unconjugated) is chiefly controlled by nuclear receptors and the uptake of long-chain fatty acids (LCFA) and BA conjugation via transporters. Among them, from in-vivo studies, farnesoid X receptor (FXR) and liver-specific fatty acid transport protein 5 (FATP5) have shown convincing evidence for their key roles in lipid homeostasis and reversal of fatty liver disease substantially. BAs have a wider range of biological effects as they are identified as modulators for FXR and FATP5 both and therefore hold a significant promise for altering the lipid content in the treatment of a metabolic disorder. BAs also have received noteworthy interest in drug delivery research due to its peculiar physicochemical properties and biocompatibility. Here, we are highlighting the connecting possibility of BAs as an agonist for FXR and antagonist for FATP5, paving an avenue to target them for designing synthetic small molecules for lipid homeostasis.

7 illus, 2 tables, 93 ref

043273   XU T, LI L, LIU Y-C, CAO W, CHEN J-S, HU S, LIU Y, LI L-Y, ZHOU H, MENG X-M, et al. (Anhui Medical Univ, Hefei, Anhui Province, 230032, China, Email: lj@ahmu.edu.cn) : CRISPR/Cas9-related technologies in liver diseases: From feasibility to future diversity. Int J Biol Sci 2020, 16(13), 2283-95.

Liver diseases are one of the leading causes of mortality in the world, mainly caused by different etiological agents, alcohol consumption, viruses, drug intoxication, and malnutrition. The maturation of gene therapy has heralded new avenues for developing effective interventions for these diseases. Derived from a remarkable microbial defense system, clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins 9 system (CRISPR/Cas9 system) is driving innovative applications from basic biology to biotechnology and medicine. Recently, the mutagenic function of CRISPR/Cas9 system has been widely adopted for genome and disease research. In this review, we describe the development and applications of CRISPR/Cas9 system on liver diseases for research or translational applications, while highlighting challenges as well as future avenues for innovation.

5 illus, 2 tables, 78 ref

043272   XIANG Y-T, ZHAO N, ZHAO Y-J, LIU Z, ZHANG Q, FENG Y, YAN X-N, CHEUNG T, NG⁶ C S (Macau Univ, Taipa, Macau SAR, China, Email: xyutly@gmail.com) : An overview of the expert consensus on the mental health treatment and services for major psychiatric disorders during COVID-19 outbreak: China's experiences. Int J Biol Sci 2020, 16(13), 2265-70.

The 2019 novel coronavirus disease (COVID-19) epidemic in China has presented substantial challenges to traditional forms of mental health service delivery. This review summarizes the expert consensus on the mental health treatment and services for severe psychiatric disorders during the COVID-19 outbreak developed by the Chinese Society of Psychiatry and other academic associations. The Expert Recommendations on Managing Patients with Mental Disorders during a Serious Infectious Disease Outbreak (COVID-19) outline the appropriate measures for psychiatric hospitals or psychiatric units in general hospitals, including the delivery of outpatient, inpatient, and community mental health services. The Expert Recommendations on Internet and Telehealth in Psychiatry during Major Public Health Crises (COVID-19) describe the assessment and treatment issues of internet-based mental health services during the COVID-19 outbreak. The expert consensus recommendations provide guidance for mental health professionals in managing psychiatric services during the COVID-19 outbreak in China. The experiences from China in addressing the challenges in the management of major psychiatric disorders may be useful and relevant to other countries who are combating the COVID-19 pandemic.

12 ref

043271   HUANG X, HE C, HUA X, KAN A, SUN S, WANG J, LI S (Pancreatobiliary Surgery Dep, Sun Yat-sen Univ Cancer Center, Guangzhou 510060, People’s Republic of China, Email: lishengp@mail.sysu.edu.cn) : Bioinformatic analysis of correlation between immune infiltration and COVID-19 in cancer patients. Int J Biol Sci 2020, 16(13), 2464-76.

In 2020, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused infections worldwide. However, the correlation between the immune infiltration and coronavirus disease 2019 (COVID-19) susceptibility or severity in cancer patients remains to be fully elucidated. ACE2 expressions in normal tissues, cancers and cell lines were comprehensively assessed. Furthermore, we compared ACE2 expression between cancers and matched normal tissues through Gene Expression Profiling Interactive Analysis (GEPIA). In addition, we performed gene set enrichment analysis (GSEA) to investigate the related signaling pathways. Finally, the correlations between ACE2 expression and immune infiltration were investigated via Tumor Immune Estimation Resource (TIMER) and GEPIA. We found that ACE2 was predominantly expressed in both adult and fetal tissues from the digestive, urinary and male reproductive tracts; moreover, ACE2 expressions in corresponding cancers were generally higher than that in matched healthy tissues. GSEA showed that various metabolic and immune-related pathways were significantly associated with ACE2 expression across multiple cancer types. Intriguingly, we found that ACE2 expression correlated significantly with immune cell infiltration in both normal and cancer tissues, especially in the stomach and colon. These findings proposed a possible fecal-oral and maternal-fetal transmission of SARS-CoV-2 and suggested that cancers of the respiratory, digestive or urinary tracts would be more vulnerable to SARS-CoV-2 infection.

5 illus, 3 tables, 45 ref

043270   LIANG J-Y, WANG D-S, LIN H-C, CHEN X-X, YANG H, ZHENG Y, LI Y-H (Medical Oncology Dep, Sun Yat-sen Univ Cancer Center, Guangzhou, People's Republic of China, Email: liyh@sysucc.org.cn) : A novel ferroptosis-related gene signature for overall survival prediction in patients with hepatocellular carcinoma. Int J Biol Sci 2020, 16(13), 2430-41.

Hepatocellular carcinoma (HCC) is a highly heterogeneous disease, which makes the prognostic prediction challenging. Ferroptosis, an iron-dependent form of regulated cell death, can be induced by sorafenib. However, the prognostic value of ferroptosis-related genes in HCC remains to be further elucidated. In this study, the mRNA expression profiles and corresponding clinical data of HCC patients were downloaded from public databases. The least absolute shrinkage and selection operator (LASSO) Cox regression model was utilized to construct a multigene signature in the TCGA cohort. HCC patients from the ICGC cohort were used for validation. Our results showed that most of the ferroptosis-related genes (81.7 %) were differentially expressed between HCC and adjacent normal tissues in the TCGA cohort. Twenty-six differentially expressed genes (DEGs) were correlated with overall survival (OS) in the univariate Cox regression analysis (all adjusted P< 0.05). A 10-gene signature was constructed to stratify patients into two risk groups. Patients in the high-risk group showed significantly reduced OS compared with patients in the low-risk group (P < 0.001 in the TCGA cohort and P = 0.001 in the ICGC cohort). The risk score was an independent predictor for OS in multivariate Cox regression analyses (HR> 1, P< 0.01). Receiver operating characteristic (ROC) curve analysis confirmed the signature's predictive capacity. Functional analysis revealed that immune-related pathways were enriched, and immune status were different between two risk groups. In conclusion, a novel ferroptosis-related gene signature can be used for prognostic prediction in HCC. Targeting ferroptosis may be a therapeutic alternative for HCC.

7 illus, 2 tables, 32 ref

043269   WANG X, ZHAO Y, FEI X, LU Q, LI Y, YUAN Y, LU C, LI C, CHEN H (Thoracic Surgery Dep, Second Military Medical Univ, Shanghai 200433, China, Email: chzchanghai@163.com) : LEF1/Id3/HRAS axis promotes the tumorigenesis and progression of esophageal squamous cell carcinoma. Int J Biol Sci 2020, 16(13), 2392- 404.

Our previous study demonstrated that lymphoid enhancer-binding factor 1 (LEF1) could promote the progression of esophageal squamous cell carcinoma (ESCC). However, the regulatory mechanism of LEF1 was not clear thoroughly. Herein, we continued to explore the downstream mechanism of LEF1 in ESCC. In this study, we applied western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry, RNA-Seq analysis, a luciferase reporter assay, chromatin immunoprecipitation (ChIP), bioinformatics analysis, and a series of functional assays in vitro and in vivo. The results demonstrated that LEF1 regulated directly the expression of Id3. Id3 was highly expressed in ESCC tissues and correlated with histologic differentiation (p=0.011), pT stage (p<0.01) and AJCC stage (p<0.01) in ESCC patients. Moreover, Id3 could serve as a prognostic factor of ESCC. By various functional experiments, overexpression of Id3 promoted the proliferation, migration, invasion, EMT, and tumorgenicity. Mechanistically, Id3 could regulate ERK/MAPK signaling pathway via activating HRAS to perform its biological function. Furthermore, activating ERK/MAPK signaling pathway promoted the expression of Id3 gene in turn, indicating that a positive regulatory loop between Id3 and ERK/MAPK pathway may exist in ESCC. In summary, LEF1/Id3/HRAS axis could promote the tumorigenesis and progression of ESCC via activating ERK/MAPK signaling pathway. Targeting this cascade may provide a valid antitumor strategy to delay ESCC progress.

8 illus, 4 tables, 38 ref

043268   XIANG Z, LIU J, SHI D, CHEN W, LI J, YAN R, BI Y, HU W, ZHU Z, YU Y, et al. (Emergency Dep, Shanghai Jiaotong Univ School of Medicine, 200025, Shanghai, China, Email: yangzhitao@hotmail.fr) : Glucocorticoids improve severe or critical COVID-19 by activating ACE2 and reducing IL-6 levels. Int J Biol Sci 2020, 16(13), 2382-91.

COVID-19 is a public health emergency that has rapidly spread to over 200 countries and regions, and no effective treatment has been established to date. Severe and critical cases have been associated with higher mortality due to acute respiratory distress syndrome (ARDS) and cytokine storm. Based on the novelty and recent emergence of COVID-19, no effective treatment regimen has been identified, thus prompting clinicians to engage in drug repurposing to address the immediate therapeutic need. This study focused on the molecular target angiotensin-converting enzyme 2 (ACE2) of SARS-CoV-2 and screened a group of ACE2 agonists by bioinformatics. Glucocorticoids are a type of ACE2 activator. We verified the efficacy of nine chemicals on regulating ACE2 expression in human GES-1, an upper digestive tract epithelial cell line, and THP-1, a human monocyte cell line, and found that several glucocorticoids imparted activating effects on ACE2 in both cell lines. The drugs triciribine and kinetin riboside activate ACE2 expression or inhibit IL-6 production in macrophages to some extent. In addition, we compared the efficacies of several glucocorticoids. Hydrocortisone showed the strongest effect on ACE2 activation, followed by prednisolone, dexamethasone, and methylprednisolone. We retrospectively analyzed the therapeutic efficacy of nine severe or critical patients from a cohort of 90 COVID-19 cases, who received medium to small doses of glucocorticoids from our integrated medical team in Wuhan. Seven out of nine patients revealed significant improvement in clinical parameters and chest CT images. This study provides experimental and clinical evidence that medium-to-low-dose glucocorticoids may play a protective role in the respiratory and digestive systems by activating ACE2 and suppressing cytokine storm.

4 illus, 2 tables, 30 ref

043267   YU T, GONG Y, LIU Y, XIA L, ZHAO C, LIU L, XIE M, WU Z, ZHAO D, QIU W, et al. (Immunology Dep, Nanjing Medical Univ, Jiangsu 211166, China, Email: zhangjing@njmu.edu.cn) : KLF6 acetylation promotes sublytic C5b-9-induced production of MCP-1 and RANTES in experimental mesangial proliferative glomerulonephritis. Int J Biol Sci 2020, 16(13), 2340-56.

Rat Thy-1 nephritis (Thy-1N) is an experimental mesangial proliferative glomerulonephritis (MsPGN) for studying human MsPGN. Although sublytic C5b-9 complex formation on glomerular mesangial cells (GMCs) and renal MCP-1 and RANTES production in rats with Thy-1N have been proved, the role and mechanism of MCP-1 or RANTES synthesis in GMCs induced by sublytic C5b-9 are poorly elucidated. In this study, we first found the expression of transcription factor (KLF6), co-activator (KAT7) and chemokines (MCP-1 and RANTES) was all up-regulated both in renal tissue of Thy-1N rats (in vivo) and in sublytic C5b-9-induced GMCs (in vitro). Further in vitro experiments revealed that KLF6 bound to MCP-1 promoter (-297 to -123 nt) and RANTES promoter (-343 to -191 nt), leading to MCP-1 and RANTES gene transcription. Meanwhile, KAT7 also bound to the same region of MCP-1 and RANTES promoter in a KLF6-dependent manner, and KLF6 was acetylated by KAT7 at lysine residue 100, which finally promoted MCP-1 and RANTES expression. Moreover, our in vivo experiments discovered that knockdown of renal KAT7 or KLF6 gene obviously reduced MCP-1 and RANTES production, GMCs proliferation, ECM accumulation, and proteinuria secretion in Thy-1N rats. Collectively, our study indicates that sublytic C5b-9-induced MCP-1 and RANTES synthesis is associated with KAT7-mediated KLF6 acetylation and elevated KLF6 transcriptional activity, which might provide a new insight into the pathogenesis of rat Thy-1N and human MsPGN.

7 illus, 47 ref

046576   PATEL J R, JOSHI H V, SHAH U A, PATEL J K (Pharmaceutical Chemistry Dep, Shri B.M. Shah Coll of Pharmaceutical Education and Research, Modasa- 383 315, Email: jimish_patel_1986@yahoo.co.in) : Exploring novel endothelin receptor blocker as anti-hypertensive agents identified from a natural drugs library using induced fit docking and biological assay. Med Plants 2020, 12(3), 405-13.

For millions of years, nature has provided many potent drugs for complicated diseases. Natural products have exhibited paramount sources of novel drugs and have gained a dominant role in drug design and discovery. Dual endothelin receptor blockers are established as novel anti-hypertensive agents in recent years. Therefore, an attempt has been made to discover molecules from different plant active chemical constituents against anti-hypertensive target. To discover the definite antihypertensive sources from natural products, in silico induced fit docking and virtual screening were implemented to obtain potential compounds, which were further identified by biological screening to check the efficiency of identified compounds with bosentan. Out of all compounds, bacoside A had a good affinity towards (Docking score; 10.7 kcal/mol) the antagonism of endothelin receptor. The docking affinity was also confirmed through biological assay. The bacoside A showed more inhibition of endothelin receptors as compared to bosentan. Therefore, our computational study suggested that bacoside A as a lead compound for further exploring more potent compounds as endothelin receptor blockers.

6 illus, 4 tables, 22 ref

043266   XIAO Z, LIU Y, ZHAO J, LI L, HU L, LU Q, ZENG Z, LIU X, HUANG D, YANG W, et al. (Zhejiang Provincial People’s Hospital (People’s Hospital of Hangzhou Medical Coll), Zhejiang 310014, China, Email: liuxin@hmc.edu.cn) : Long noncoding RNA LINC01123 promotes the proliferation and invasion of hepatocellular carcinoma cells by modulating the miR-34a-5p/TUFT1 axis. Int J Biol Sci 2020, 16(13), 2296-305.

Hepatocellular carcinoma (HCC), one of the main causes of cancer-related deaths globally, is characterized by rapid growth and high invasiveness. Accumulating evidence demonstrates that long noncoding RNAs (lncRNAs) play critical roles in the growth and metastasis of HCC. Recently, lncRNA LINC01123 has been found to contribute to cell proliferation and aerobic glycolysis in lung cancer. However, the function of LINC01123 in HCC, as well as the underlying mechanism of its action, remain unclear. Here, we found that the expression of LINC01123 was clearly upregulated in HCC tissues compared to nontumor tissues. Furthermore, expression of LINC01123 in HCC cells was significantly higher than in LO2 cells. Importantly, the upregulated level of LINC01123 was related to unfavorable clinical features and poor prognosis of HCC. Next, we demonstrated that LINC01123 knockdown suppressed the proliferation, migration and invasion of HCC cells in vitro. Depletion of LINC01123 inhibited HCC xenograft growth in vivo. Conversely, ectopic expression of LINC01123 facilitated HCC cell proliferation and invasion. Mechanistically, LINC01123 acted as a molecular sponge for miR-34a-5p in HCC cells. Tuftelin1 (TUFT1) was identified as the target gene of miR-34a-5p. LINC01123 positively regulated TUFT1 level by targeting of miR-34a-5p in HCC cells. Notably, TUFT1 restoration can abolish miR-34a-5p-induced inhibitory effects on HCC cell proliferation, migration and invasion. In conclusion, LINC01123 was overexpressed in HCC and accelerated cancer cell proliferation and invasion by regulating the miR-34a-5p/TUFT1 axis.

7 illus, 1 table, 33 ref

043265   LI C, CAO Y, ZHANG L, LI J, WU H, LING F, ZHENG J, WANG J, LI B, HE J, et al. (Southern Medical Univ, Foshan, China, Email: zhang_40_1@163.com) : LncRNA IGFBP4-1 promotes tumor development by activating Janus kinase-signal transducer and activator of transcription pathway in bladder urothelial carcinoma. Int J Biol Sci 2020, 16(13), 2271-82.

Insulin-like growth factor binding protein 4–1 (IGFBP4–1), a new long noncoding RNA (lncRNA), has been reported to contribute to tumorigenesis and has been suggested to be a poor prognostic marker in human lung cancer. However, there still lacks basic studies that investigated the biological role of IGFBP4–1 in bladder urothelial carcinoma to date. In this study, we investigated the relationship between IGFBP4–1 expression and prognosis in patients with bladder cancer. Cell proliferation, cell cycle and cell apoptosis assays were performed to assess IGFBP4–1 function by up-regulating or down-regulating IGFBP4–1 in bladder cancer cells. A xenograft mice model was used to validate the in vitro results. Blockade of Janus kinase-signal transducer and activator of transcription pathway (JAK/STAT) was used to evaluate JAK/STAT signaling activity. The results showed that IGFBP4–1 was overexpressed in bladder cancer tissues compared with that in normal bladder tissues, and its expression level was positively correlated with poor prognosis in bladder cancer patients. Overexpression of IGFBP4–1 markedly promoted cell proliferation and cell cycle progression, and inhibited cell apoptosis, while knockdown of IGFBP4–1 notably suppressed the proliferation, promoted cell apoptosis, and induced cell cycle arrest at the G0/G1 phase. Mechanistically, we revealed that IGFBP4–1 promotes the activation of the JAK/STAT pathway in bladder cancer cells. Moreover, the JAK/STAT inhibitor dramatically blocked the tumor-promoting activity of IGFBP4–1. Tumor growth in vivo was also suppressed by knocking down of IGFBP4–1. In conclusion, IGFBP4–1 promoted bladder cancer progression by activating the JAK/STAT signaling pathway. These findings suggest that IGFBP4–1 exhibits an oncogenic role in the development of human bladder cancer.

6 illus, 3 tables, 32 ref

043264   SHI J-W, LAI Z-Z, YANG H-L, YANG S-L, WANG C-J, AO D, RUAN L-Y, SHEN H-H, ZHUA W-J, MEI J, et al. (Fudan Univ, Shanghai 200080, People’s Republic of China, Email: mqli@fudan.edu.cn) : Collagen at the maternal-fetal interface in human pregnancy. Int J Biol Sci 2020, 16(12), 2220-34.

The survival and development of a semi-allogenic fetus during pregnancy require special immune tolerance microenvironment at the maternal fetal interface. During the establishment of a successful pregnancy, the endometrium undergoes a series of changes, and the extracellular matrix (ECM) breaks down and remodels. Collagen is one of the most abundant ECM. Emerging evidence has shown that collagen and its fragment are expressed at the maternal fetal interface. The regulation of expression of collagen is quite complex, and this process involves a multitude of factors. Collagen exerts a critical role during the successful pregnancy. In addition, the abnormal expressions of collagen and its fragments are associated with certain pathological states associated with pregnancy, including recurrent miscarriage, diabetes mellitus with pregnancy, preeclampsia and so on. In this review, the expression and potential roles of collagen under conditions of physiological and pathological pregnancy are systematically discussed.

2 illus, 2 tables, 152 ref

041462   PHERWANI P U, SATHAYE S (Pharmaceutical Sciences and Technology Dep, Institute of Chemical Technology (ICT), Mumbai - 400 019, Maharashtra, Email: sadhanasathaye@hotmail.com) : Treatment of osteoporosis: Current scenario from a research perspective. Indian J Pharm Edu Res 2020, 54(1), 8-16.

Osteoporosis is a condition of compromised bone quality and bone density increasing the chances of fracture. In osteoporotic bone, there is a mismatch between bone formation and resorption. The high probability in both women (33.33 %) and males (20 %) over the age of 50 worldwide to suffer a fracture draws attention to the importance of treatment of osteoporosis. The benefits of using bisphosphonates, the first line of therapy in osteoporosis lasts for 5 years. The other drugs are recommended for a still shorter time. The state of usage of current drugs is discussed. Two drugs Abaloparatide and Romosozumab have been recently approved. There is a better understanding of the means of communication amongst the different players in bone remodeling. These players involved in remodeling, are subjected to local and systemic influences. This increased understanding of physiology led to molecules being investigated which acts on newer targets such as Cathepsin K, ɑ_v β₃ integrin, chloride channel 7, tryptophan hydroxylase 1, Src, calcium-sensing receptor. Also, a bone anabolic effect for certain drugs belonging to different therapeutic classes such as statins, nitrates, thiazides, beta-blockers has been noted. However, for these drugs, further studies regarding the dose and frequency of drug administration and the effectiveness in the prevention of fractures along with safety profiles are needed. The drugs acting on new targets and new uses of old drugs hold hope to increase our arsenal against osteoporosis and thus preventing the mortality and debility due to fractures.

2 illus, 1 table, 50 ref

041461   CHANDRASEKAR K, SAI N G, JOHN P S, NINAN S, DURAI R, PONNUSANKAR S (Pharmacy Practice Dep, JSS Coll of Pharmacy, Ooty - 643 001, Tamil Nadu, Email: ponnusankarsivas@gmail.com) : Emerging non-pharmacological therapies for post-stroke depression and its future aspects: A review. Indian J Pharm Edu Res 2020, 54(1), 1-7.

Post-Stroke Depression (PSD) is a psychiatric disorder associated with stroke which has an adverse effect on the cognitive function and survival. It usually develops in 40 % of the stroke survivors within 3 months. We performed a thorough literature review using PsychInfo, PubMed, Science Direct and PLOS databases for the non-pharmacological treatment of PSD. Early rehabilitation and psychological therapies are effective in treating depression in PSD while physiotherapy and music therapy improves both cognitive and movement disability. Transcranial stimulation and electroconvulsive therapy are invasive procedures used to treat cognitive impairment in PSD. Computerized therapy helps, enhance the attention, memory and executive functioning. DepReT stroke is a package which addresses patients along with their caregiver to have a better understanding of the depression. Robot assisted rehabilitation along with traditional rehabilitation is effective in treating motor function associated with PSD. Early assessment, treatment and rehabilitation are the most effective ways to prevent depression in stroke survivors and thereby improving their quality of life. Non-pharmacological treatment for post-stroke depression seem to be efficacious as antidepressants in improving depressive symptoms with minimal side effects.

1 illus, 54 ref

043263   ZHOU L, ZHU Y, SUN D, ZHANG Q (Taizhou Univ, Zhejiang, China, Email: sunds@tzc.edu.cn) : Emerging roles of long non-coding RNAs in the tumor microenvironment. Int J Biol Sci 2020, 16(12), 2094-03.

Long non-coding RNAs (lncRNAs) are a diverse class of longer than 200 nucleotides RNA transcripts that have limited protein coding capacity. LncRNAs display diverse cellular functions and widely participate in both physiological and pathophysiological processes. Aberrant expressions of lncRNAs are correlated with tumor progression, providing sound rationale for their targeting as attractive anti-tumor therapeutic strategies. Emerging evidences support that lncRNAs participate in tumor-stroma crosstalk and stimulate a distinctive and suitable tumor microenvironment (TME). The TME comprises several stromal cells such as cancer stem cells (CSCs), cancer-associated endothelial cells (CAEs), cancer-associated fibroblasts (CAFs) and infiltrated immune cells, all of which are involved in the complicated crosstalk with tumor cells to affect tumor progression. In this review, we summarize the essential properties and functional roles of known lncRNAs in related to the TME to validate lncRNAs as potential biomarkers and promising anti-cancer targets.

3 illus, 2 tables, 97 ref

041460   KAUR S, KAUR H, KOMAL K, KAUR P, KAUR D, JARIYAL V L, KAVITA K, BAJAJ L, THAKUR J S (National Institute of Nursing Education, Chandigarh - 160 012, Email: gaurikavita@rediffmail.com) : Need of palliative care services in rural area of Northern India. Indian J Palliat Care 2020, 26(4), 528-30.

Palliative care is comprehensive care that provides symptomatic relief and enhances the quality of life for people experiencing serious health‑related suffering. There is an increasing need for palliative care services in India. Estimates for population requiring these services are essential in order to meet the increasing need for palliative care services. The objective was to assess the proportion of population requiring and receiving palliative care services. A descriptive cross‑sectional research design was adopted for the study. The study was carried out in the village, Dhanas, Chandigarh. All the residents residing in the selected rural area were included in the study. Tools used for data collection were a screening questionnaire consisting of three questions, a sociodemographic sheet, a clinical profile, the Barthel Index of activities of daily living (ADL), and a pain rating scale. Analysis of the data was done using SPSS version 19 (IBM SPSS Statistics for Windows, Version 19.0.: IBM Corp). Atotal of 10,021 people from 884 households were screened in the study. The results revealed that the prevalence of need for palliative care services was 2/1000 population. None of the 19 participants with unmet palliative care needs were receiving any home‑ or institutional‑based palliative care services at the time of assessment. Nearly, one‑fourth of the participants had total dependency on caregivers for ADL. The study concluded that there is a need of palliative care services in the studied rural community of Chandigarh. These data can be used for planning and implementing community‑based palliative care services in the studied area.

2 tables, 15 ref

043262   HU X, CHEN H, XU H, WU Y, WU C, JIA C, LI Y, SHENG S, XU C, XU H, et al. (Orthopaedics Dep, Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical Univ, Wenzhou 325000, Zhejiang, China, Email: wenfeini@yeah.net) : Role of pyroptosis in traumatic brain and spinal cord injuries. Int J Biol Sci 2020, 16(12), 2042-50.

Central nervous system (CNS) trauma, including traumatic brain injury (TBI) and spinal cord injury (SCI), remains a leading cause for morbidity and mortality worldwide. Past research has shown that cell death plays a critical role in the pathophysiology of CNS injuries. More recently, pyroptosis has been identified as a form of programmed inflammatory cell death, and it is a unique form of cell death in various aspects. Mechanistically, pyroptosis can be categorized into canonical (mediated by caspase-1) and non-canonical (mediated by caspase-4/5/11). In canonical pyroptosis, Nod-like receptors (NLRs) inflammasomes play a critical role, and their activation promotes the maturation and secretion of the inflammatory cytokines interleukin-1β/18 (IL-1β/18), cleavage of gasdermin D (GSDMD), and ultimately pyroptotic cell death. Despite a plethora of new knowledge regarding pyroptosis, detailed understanding of how pyroptosis is involved in CNS injuries and possible ways to improve clinical outcomes following CNS injuries remain elusive. This review discusses the current knowledge on how pyroptosis is involved in CNS injuries, focusing on new discoveries regarding how pyroptosis activation occurs, differences between CNS cell types following injury, time-course of inflammatory responses, and key regulatory steps of pyroptosis. In addition, we highlight various investigational agents that are capable of regulating key steps in pyroptotic cell death, and we discuss how these agents may be used as therapies to improve outcomes following CNS trauma.

2 illus, 1 table, 87 ref

041459   AGARWAL S, GARG R, MINHAS V, BHATNAGAR S, MISHRA S, KUMAR V, BHARATI S J, GUPTA N, KHAN M A (Onco?Anaesthesia and Palliative Medicine Dep, All India Institute of Medical Sciences, Ansari Nagar, New Delhi ? 110 029, Email: drrgarg@hotmail.com) : To assess the prevalence and predictors of cancer related fatigue and its impact on quality of life in advanced cancer patients receiving palliative care in a tertiary care hospital: A cross sectional descriptive study. Indian J Palliat Care 2020, 26(4), 523-7.

Cancer‑related fatigue (CRF) is one of the adverse outcomes of cancer and its treatment. Despite its high prevalence; the data are scarce from the Indian population on the prevalence of CRF and its predictors in advanced cancer patients. Hence, we aim to find the prevalence of the fatigue, its impact of fatigue on quality of life (QOL), and possible predictors. This study was conducted after approval of the ethical committee in adult patients of advanced cancer receiving palliative care. The data collected included demographic details, nutritional status, any comorbidities involving cardiorespiratory, renal, pulmonary, and neurological system, type and stage of cancer, site of metastasis, any previous or ongoing chemotherapy or radiotherapy, history of drug intake, hemoglobin, and albumin. The study parameters included assessment of fatigue, QOL, and symptom assessment as per the validated tools. The primary objective of the study was to find the prevalence of fatigue in advanced cancer patients receiving palliative care. The secondary objectives were to find predictive factors of fatigue, its impact on QOL of patients, and the relation between the fatigue and QOL receiving palliative care. The correlation between fatigue score and QOL was analyzed using Pearson’s correlation coefficient. Multiple linear regression analysis was performed for identifying the predictors of CRF. The fatigue was observed in all 110 patients in this study. Of these, severe fatigue was seen in 97 patients (Functional Assessment of Chronic Illness Therapy [FACIT]‑F < 30). The median (interquartile range [IQR]) FACIT‑F score was 14 (8–23). The median (IQR) of the overall QOL was 16.66 (16.6–50).The correlation between the fatigue (FACIT‑F) and QOL was + 0.64 (P < 0.001). The predictors of fatigue included pain, physical functioning, Eastern Cooperative Oncology Group, tiredness, and the level of albumin. We conclude that the prevalence of fatigue in Indian patients with advanced cancer receiving palliative care was high and it has a negative impact on QOL. Pain, physical functioning, performance status, and albumin were found to be independent predictors of CRF.

6 tables, 32 ref

041458   ALSAATI B A, ALJISHI M N, ALSHAMAKH S S, BASHARAHEEL H A, BANJAR N S, ALAMRI R S, ALKHAYYAT S (Medicine Dep, King Abdul-Aziz Univ, Jeddah, Saudi Arabia., Email: bashayer.alsaati@hotmail.com) : The concept of do not resuscitate for the families of the patients at king abdul-aziz university hospital. Indian J Palliat Care 2020, 26(4), 518-22.

Do not resuscitate (DNR) is an order in medical practice for the patients who are suffering from a grave medical condition, and their life is in danger to end. DNR decision-making varies from one hospital to another. This study is aimed to assess the knowledge of the patients’ relatives about DNR concept and their opinion about the DNR decision-making. This was a nonintervention cross-sectional study conducted, during 2016, among 420 patients’ relatives in the Emergency Department at King Abdul-Aziz University Hospital in KSA. Data were collected by interviewing the participants. Data were further analyzed using SPSS software. The Chi-square test was used to determine the associations. Variation in responses related to the DNR concept was observed. Around 44 % of participants thought that DNR involved maximum intervention in the hospital, including intensive care. Further, the majority (55.2 %) of the participants were assured about the quality of the services the patient would receive. Furthermore, 51 % of the participants believed that ultimately, it should always be the doctor who decides on a DNR decision. Meanwhile, 36.4 % of the relatives opined that the family members should be involved in the discussion regarding the DNR order. We observed a gap in the understanding of the concept and decision-making of DNR-order among the participants. Health-care providers should provide a greater explanation about DNR orders to the families of the patients to avoid any misunderstandings, and also support them psychologically to avoid any stress they might encounter in such situations.

1 illus, 7 tables, 17 ref

043261   ZHU X, XUAN Z, CHEN J, LI Z, ZHENG S, SONG P (Key Laboratory of Organ Transplantation, Hangzhou 310003, China, Email: songpenghong@zju.edu.cn) : How DNA methylation affects the warburg effect. Int J Biol Sci 2020, 16(12), 2029-41.

Significant enhancement of the glycolysis pathway is a major feature of tumor cells, even in the presence of abundant oxygen; this enhancement is known as the Warburg effect, and also called aerobic glycolysis. The Warburg effect was discovered nearly a hundred years ago, but its specific mechanism remains difficult to explain. DNA methylation is considered to be a potential trigger for the Warburg effect, as the two processes have many overlapping links during tumorigenesis. Based on a widely recognized potential mechanism of the Warburg effect, we here summarized the relationship between DNA methylation and the Warburg effect with regard to cellular energy metabolism factors, such as glycolysis related enzymes, mitochondrial function, glycolysis bypass pathways, the tumor oxygen sensing pathway and abnormal methylation conditions. We believe that clarifying the relationship between these different mechanisms may further help us understand how DNA methylation works on tumorigenesis and provide new opportunities for cancer therapy.

3 illus, 164 ref

041457   AGARWAL A, GAUTAM A, RASTOGI S, MALVIYA D, DAS P K, HARJAI M (Anesthesiology Dep, Dr RMLIMS, Lucknow, Uttar Pradesh, Email: neuromanu@yahoo.com) : Effect of celiac plexus neurolysis for pain relief in patients with upper abdominal malignancy: A retrospective observational study and review of literature. Indian J Palliat Care 2020, 26(4), 512-7.

Abdominal pain from primary cancer or metastatic disease is a significant cause of pain for patients undergoing treatment for the disease. Patient’s pain may be resistant or non-responsive to the pharmacological management, hence minimal invasive pain intervention like celiac plexus neurolysis or splanchnic nerve rhizolysis may be required to relieve pain of such patients. The aim of this retrospective study is to assess the effect of celiac plexus neurolysis for pain relief in patients with upper gastro-intestinal malignancies. This is a retrospective, observational study with short review. This retrospective observational study was done in the Pain Medicine unit from 2016 and November 2018. Ninety-four patients with upper abdominal malignancy and unrelenting pain, non-responsive or poorly responsive to pharmacological treatment as per WHO ladder of analgesics, received fluoroscopy-guided celiac plexus neurolysis (CPN). All the patients underwent celiac plexus neurolysis through Trans-Aortic approach and the primary outcome measure was pain as assessed with Visual Analogue Scale (VAS) ranging from 0 to 10; the secondary outcome measures were morphine consumption per day (M), quality of life (QOL) as assessed by comparing the percent of positive responses and complications, if any. These were noted and analyzed prior to intervention and then on day 1, and months 1, 2, 3, 4, 5, 6 following CPN. Follow up was completed 6 months after the procedure. VAS score, daily morphine consumption, and the quality of life showed improvement for the duration of the study. There was some relapse in pain and deterioration in QOL during the fourth to sixth month of pain intervention due to disease progression. Some transient known side effects also occurred. Trans-Aortic celiac plexus neurolysis with low volume of alcohol is a safe procedure providing up to 6 months of pain relief and is an effective, well established, minimally invasive procedure for abdominal pain due to primary malignancy or metastatic spread.

5 illus, 4 tables, 21 ref

043260   LI X, WANG J (Obstetrics and Gynecology Dep, Peking Univ People's Hospital, Xicheng District, Beijing 100044, China, Email: wangjianliu@pkuph.edu.cn) : Mechanical tumor microenvironment and transduction: cytoskeleton mediates cancer cell invasion and metastasis. Int J Biol Sci 2020, 16(12), 2014-28.

Metastasis is a complicated, multistep process that is responsible for over 90% of cancer-related death. Metastatic disease or the movement of cancer cells from one site to another requires dramatic remodeling of the cytoskeleton. The regulation of cancer cell migration is determined not only by biochemical factors in the microenvironment but also by the biomechanical contextual information provided by the extracellular matrix (ECM). The responses of the cytoskeleton to chemical signals are well characterized and understood. However, the mechanisms of response to mechanical signals in the form of externally applied force and forces generated by the ECM are still poorly understood. Furthermore, understanding the way cellular mechanosensors interact with the physical properties of the microenvironment and transmit the signals to activate the cytoskeletal movements may help identify an effective strategy for the treatment of cancer. Here, we will discuss the role of tumor microenvironment during cancer metastasis and how physical forces remodel the cytoskeleton through mechanosensing and transduction.

3 illus, 191 ref

043259   ZHU X, LIU H, ZHANG Z, WEI R, ZHOU X, WANG Z, ZHAO L, GUO Q, ZHANG Y, CHU C, et al. (Obstetrics and Gynecology Dep, First Affiliated Hospital of Shandong First Medical Univ, Jinan 250014, Shandong, China, Email: wangli1415@sdhospital.com.cn) : MiR-103 protects from recurrent spontaneous abortion via inhibiting STAT1 mediated M1 macrophage polarization. Int J Biol Sci 2020, 16(12), 2248-64.

Recurrent spontaneous abortion (RSA) is a common complication of early pregnancy. Excessive M1 macrophage was found to be involved in RSA, but the underlying mechanisms remains unclear. MicroRNAs play critical roles in RSA as well as the polarization of macrophages; however, the regulatory effect of miRNAs on M1 differentiation in RSA has not been fully investigated. In this study, miRNA microarray assay revealed that miR-103 was significantly decreased in RAW264.7-derived M1 macrophages upon IFNγ and LPS stimulation. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that in RSA patients, miR-103 expression was decreased substantially, and negatively correlated with that of STAT1. Moreover, down-regulation of miR-103 could sensitively discriminate RSA patients from normal pregnancies (NP) subjects. Experiments in vitro showed that overexpression of miR-103 suppressed M1 polarization by inhibiting STAT1/IRF1 signaling pathway and vice versa. miR-103 regulated STAT1 expression by direct binding to its 3’-UTR. Moreover, our in vivo study demonstrated that overexpressed miR-103 could reduce mice embryo resorption and M1 polarization effectively. Overall, the results suggested that decreased miR-103 was involved in RSA by increasing M1 macrophage polarization via promoting STAT1/IRF1 signaling pathway. miR-103 may be explored as a promising diagnostic marker and therapeutic target for RSA.

8 illus, 59 ref

043258   TANG L, GAO Y, SONG Y, LI Y, LI Y, ZHANG H, LI D, LI J, LIU C, LI F (China Medical Univ, Shenyang, 110122, Liaoning, China, Email: lifeng@cmu.edu.cn) : PAK4 phosphorylating RUNX1 promotes ERα-positive breast cancer-induced osteolytic bone destruction. Int J Biol Sci 2020, 16(12), 2235-47.

The biological function of nuclear PAK4 in ERα-positive breast cancer osteolytic bone destruction remains unclear. Here, we find that the nuclear PAK4 promotes osteoclastogenesis and tumor-induced osteolysis via phosphorylating RUNX1. We show that nuclear PAK4 interacts with and phosphorylates RUNX1 at Thr-207, which induces its localization from the nucleus to the cytoplasm and influences direct interaction with SIN3A/HDAC1 and PRMT1. Furthermore, we reveal that RUNX1 phosphorylation by PAK4 at Thr-207 promotes osteolytic bone destruction via targeting downstream genes related to osteoclast differentiation and maturation. Importantly, we verify changes in RUNX1 subcellular localization when nuclear PAK4 is positive in breast cancer bone metastasis tissues. Functionally, we demonstrate that RUNX1 phosphorylation promotes osteolytic bone maturation and ERα-positive breast cancer-induced osteolytic bone damage in the mouse model of orthotopic breast cancer bone metastasis. Our results suggest PAK4 can be a therapeutic target for ERα-positive breast cancer osteolytic bone destruction.

6 illus, 56 ref

043257   CHEN Z, YANG X, BI G, LIANG J, HU Z, ZHAO M, LI M, LU T, ZHENG Y, SUI Q, et al. (Thoracic Surgery Dep, Fudan Univ, No. 180, Fenglin Road, Shanghai, 200032, China, Email: czhan10@fudan.edu.cn) : Ligand-receptor interaction atlas within and between tumor cells and T cells in lung adenocarcinoma. Int J Biol Sci 2020, 16(12), 2205-19.

Lung adenocarcinoma (LUAD) is the leading cause of cancer-related deaths worldwide. Although tumor cell–T cell interactions are known to play a fundamental role in promoting tumor progression, these interactions have not been explored in LUAD. The 10x genomics single-cell RNA sequencing (scRNA-seq) and gene expression data of LUAD patients were obtained from ArrayExpress, TCGA, and GEO databases. scRNA-seq data were analyzed and infiltrating tumor cells, epithelial cells, and T cells were identified in the tumor microenvironment. Differentially expressed ligand-receptor pairs were identified in tumor cells/normal epithelial cells and tumor T cells/non-tumor T cells based on corresponding scRNA-seq and gene expression data, respectively. These important interactions inside/across cancer cells and T cells in LUAD were systematically analyzed. Furthermore, a valid prognostic machine-learning model based on ligand-receptor interactions was built to predict the prognosis of LUAD patients. Flow cytometry and qRT-PCR were performed to validate the significantly differently expressed ligand-receptor pairs. Overall, 39,692 cells in scRNA-seq data were included in our study after quality filtering. A total of 65 ligand-receptor pairs (17 upregulated and 48 downregulated), including LAMB1-ITGB1, CD70-CD27, and HLA-B-LILRB2, and 96 ligand-receptor pairs (41 upregulated and 55 downregulated), including CCL5-CCR5, SELPLG-ITGB2, and CXCL13-CXCR5, were identified in LUAD cancer cells and T cells, respectively. To explore the crosstalk between cancer cells and T cells, 114 ligand-receptor pairs, including 11 ligand-receptor pair genes that could significantly affect survival outcomes, were identified in our research. A machine-learning model was established to accurately predict the prognosis of LUAD patients and ITGB4, CXCR5, and MET were found to play an important role in prognosis in our model. Flow cytometry and qRT-PCR analyses indicated the reliability of our study. Our study revealed functionally significant interactions within and between cancer cells and T cells. We believe these observations will improve our understanding of potential mechanisms of tumor microenvironment contributions to cancer progression and help identify potential targets for immunotherapy in the future. 

10 illus, 55 ref

043256   XIA X, HE J, LIU B, SHAO Z, XU Q, HU T, YU C, LIU X, LIAO Y, LIU N, et al. (Second Affiliated Hospital of Guangzhou Medical Univ, Guangdong, 510260, China, Email: liuningning@gzhmu.edu.cn ) : Targeting ERα degradation by L-tetrahydropalmatine provides a novel strategy for breast cancer treatment. Int J Biol Sci 2020, 16(12), 2192-204.

The incidence and mortality of breast cancer (BCa) are the highest among female cancers. There are approximate 70 % BCa that are classified as estrogen receptor alpha (ERα) positive. Therefore, targeting ERα is the most significantly therapeutic schedule. However, patients with breast cancer develop resistance to ERα or estrogen (E2) antagonists such as fulvestrant and tamoxifen. In the present study, we found that L-Tetrahydropalmatine (L-THP) significantly suppressed cell proliferation in ERα+ BCa cells via inducing cell cycle arrest rather than apoptosis. Additionally, L-THP enhanced the sensitivity of ERα+ BCa cells to tamoxifen and fulvestrant. Mechanically, the application of L-THP promotes ERα degradation through accumulating ubiquitin chains on ERα. Overexpressing ERα abrogates L-THP induced-antiproliferation in ERα+ BCa cells. Collectively, our work indicates that L-THP may represent a potentially novel therapeutic medicine for ERα+ breast cancer patient.

8 illus, 37 ref

043255   YING L, FEI X, JIALUN L, JIANPENG X, JIE W, ZHAOLIN M, HONGJIA F, HUAN F, SHA L, QIUJU W, et al. (Laboratory Medicine & Central Laboratory Dep, Southern Medical Univ Affiliated Fengxian Hospital, Shanghai 201499, China, Email: fengjing8801530@163.com) : SETDB2 promoted breast cancer stem cell maintenance by interaction with and stabilization of ΔNp63α protein. Int J Biol Sci 2020, 16(12), 2180-91.

The histone H3K9 methyltransferase SETDB2 is involved in cell cycle dysregulation in acute leukemia and has oncogenic roles in gastric cancer. In our study, we found that SETDB2 plays essential roles in breast cancer stem cell maintenance. Depleted SETDB2 significantly decreased the breast cancer stem cell population and mammosphere formation in vitro and also inhibited breast tumor initiation and growth in vivo. Restoring SETDB2 expression rescued the defect in breast cancer stem cell maintenance. A mechanistic analysis showed that SETDB2 upregulated the transcription of the ΔNp63α downstream Hedgehog pathway gene. SETDB2 also interacted with and methylated ΔNp63α, and stabilized ΔNp63α protein. Restoring ΔNp63α expression rescued the breast cancer stem cell maintenance defect which mediated by SETDB2 knockdown. In conclusion, our study reveals a novel function of SETDB2 in cancer stem cell maintenance in breast cancer.

7 illus, 51 ref

041456   SHAHEEN N A, ALQAHTANI M, ALAWBTHANI N S, THOMAS A, ALASKAR A (Biostatistics and Bioinformatics Dep, King Saud bin Abdulaziz Univ for Health Sciences, Riyadh 11426, Kingdom of Saudi Arabia, Email: drnaila@hotmail.com) : Chemotherapy induced peripheral neuropathy and its impact on health related quality of life among multiple myeloma patients: A single center experience. Indian J Palliat Care 2020, 26(4), 506-11.

Chemotherapy‑induced peripheral neuropathy (CIPN) is a long‑term neurological health issue in patients diagnosed with multiple myeloma (MM). The aim of this study was to assess CIPN symptoms and health‑related quality of life (HRQOL) among MM patients. A cross‑sectional survey was conducted among patients diagnosed with MM in a tertiary care hospital using a self‑reported Arabic questionnaire, European Organization for Research and Treatment of Cancer Quality‑of‑Life Questionnaire for CIPN scale (QLQ‑CIPN20). The HRQOL was assessed using EORTC multiple myeloma module (QLQ-MY20). Categorical variables were reported in frequency tables and percentages. Age and duration of MM diagnosis were reported as mean and standard deviation. Survey responses were presented using descriptive statistics. In total, 62 patients had participated. Males were 60 %. The average age was 58.74 ± 11.49 years. On sensory scale, 20 % reported “quite a bit”/“very much” tingling in fingers/hands, 23 % in toes/feet, 39 % numbness in fingers/hands, 37 % in toes/feet, and 43 % reported trouble standing or walking. On motor scale, 40 % reported trouble walking and 60 % had difficulty in climbing stairs/standing up from chair. On autonomic scale, 27 % reported orthostatic hypotension and only 13/37 (46 %) males reported erectile dysfunction. For HRQOL, 50 % reported bone aches/pain, 42 % reported back pain, 57 % reported feeling ill, 33 % reported lost hair, 35 % had been thinking about their illness, whereas 28 % were worried about future health and 22 % had reported being worried about dying. MM patients encounter CIPN symptoms with impaired HRQOL. Capturing CIPN as a patient‑reported outcome needs to be considered in routine clinical practice.

2 illus, 3 tables, 35 ref