043318   DAI Y-J, ZHANG W-N, WANG W-D, HE S-Y, LIANG C-C, WANG D-W (Collaborative Innovation Center for Cancer Medicine, 651 Dongfeng East Road, Guangzhou, 510060, China, Email: daiyj@sysucc.org.cn) : Comprehensive analysis of two potential novel SARS-CoV-2 entries, TMPRSS2 and IFITM3, in healthy individuals and cancer patients. Int J Biol Sci 2020, 16(15), 3028-36.

Coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2, with acute respiratory failure as the most significant symptom, has led to a global pandemic. Angiotensin-converting enzyme 2 (ACE2) is considered as the most important receptor of SARS-CoV-2 and wildly expressed in human tissues. Whereas, the extremely low expression of ACE2 in lung could hardly interpret the severe symptom of pneumonia in COVID-19 patients. Here we profiled two SARS-CoV-2 infection related genes, the transmembrane serine protease 2 (TMPRSS2) and the interferon-inducible transmembrane protein 3 (IFITM3), in human tissues and organs. Consistent with the expression and distribution of ACE2, TMPRSS2 was also highly expressed in digestive, urinary and reproductive systems, but low expressed in lung. Notably, the anti-virus protein IFITM3 also expressed much lower in lung than other tissues, which might be related to the severe lung symptoms of COVID-19. In addition, the low expression of IFITM3 in immune cells suggested that SARS-CoV-2 might attack lymphocytes and induce the cytokine release syndrome (CRS). Furthermore, cancer patients were considered as more susceptible to SARS-CoV-2 infection. Our data supposed that fourteen types of tumors might have different susceptibility to the virus according to ACE2, TMPRSS2 and IFITM3 expression patterns. Interestingly the prognosis of six types of cancers including breast carcinoma (BRCA), lung adenocarcinoma (LUAD), uterine corpus endometrial carcinoma (UCEC), renal clear cell carcinoma (KIRC), prostate adenocarcinoma (PRAD), and hepatocellular carcinoma (LIHC) were closely related to these gene expressions. Our study explored the expression and distribution profiles of two potential novel molecules that might participate in SARS-CoV-2 infection and involved in immunity, which may provide a functional basis for preventing infection of SARS-CoV-2.

5 illus, 45 ref

043317   LONG Q, HUA Y, HE L, ZHANG C, SUI S, LI Y, QIU H, TIAN T, AN X, LUO G, et al. (Cancer Center Univ, Guangzhou, China, Email: chenmiao@sysucc.org.cn) : Poly(U) binding splicing factor 60 promotes renal cell carcinoma growth by transcriptionally upregulating telomerase reverse transcriptase. Int J Biol Sci 2020, 16(15), 3002-17.

Abnormal transcriptional upregulation of telomerase reverse transcriptase (TERT) plays a dominant role in telomerase activation in various cancers. TERT promoter mutations (TPMs) have been identified as a key mechanism in TERT upregulation. However, the mechanism of TERT upregulation in cancers with low frequency of TPMs are not fully elucidated so far. The expression of PUF60 and TERT was detected by real-time PCR, western blot and immunohistochemistry. TERT promoter binding proteins were identified by streptavidin-agarose pulldown assay and mass spectrum (MS) analysis. The role of PUF60/TERT in renal cancer was evaluated on cell growth in vitro and in vivo. In this study, we identify the regulation mechanism of TERT in renal cell carcinoma (RCC) cells which have rare TPMs but exert significant upregulation of TERT. We found that TERT was highly expressed in RCC tumor tissues, and elevated TERT expression was associated with poor prognosis for patients. We also detected the relatively rare TPM status in both RCC tumor tissues and RCC cell lines. Mechanistically, PUF60, a RNA binding protein, was identified as a novel TERT regulator which bound to the TERT and transcriptionally upregulated TERT expression in RCC cells. The in vitro and in vivo experiments also demonstrated that PUF60 could promote RCC cell growth through activation of TERT expression in a TPM status independent way. Furthermore, we showed that there was a strong correlation of the expression of PUF60 and TERT in RCC tumor tissues and RCC cell lines, and the patients with high expression of PUF60 and TERT had significantly shorter survival. Collectively, these results indicated that PUF60 transcriptionally upregulated TERT expression to promote RCC growth and progression in a TPM status independent way, suggesting that the PUF60/TERT signaling pathway may serve as potential prognostic biomarkers and therapeutic targets for RCC.

6 illus, 2 tables, 52 ref

043316   PENG Q, XU H, XIAO M, WANG L (Gastroenterology and Research Center of Clinical Medicine Dep, Affiliated Hospital of Nantong Univ, Nantong 226001, Jiangsu, China, Email: xmb73@163.com) : The small molecule PSSM0332 disassociates the CRL4A^DCAF8 E3 ligase complex to decrease the ubiquitination of NcoR1 and inhibit the inflammatory response in a mouse sepsis-induced myocardial dysfunction model. Int J Biol Sci 2020, 16(15), 2974-88.

Sepsis-induced myocardial dysfunction (SIMD) is a life-threatening complication caused by inflammation, but how it is initiated is still unclear. Several studies have shown that extracellular high mobility group box 1 (HMGB1), an important cytokine triggering inflammation, is overexpressed during the pathogenesis of SIMD, but the underlying mechanism regarding its overexpression is still unknown. Herein, we discovered that CUL4A (cullin 4A) assembled an E3 ligase complex with RBX1 (ring-box 1), DDB1 (DNA damage-binding protein 1), and DCAF8 (DDB1 and CUL4 associated factor 8), termed CRL4A^DCAF8, which ubiquitinated and degraded NcoR1 (nuclear receptor corepressor 1) in an LPS-induced SIMD mouse model. The degradation of NcoR1 failed to form a complex with the SP1 transcription factor, leading to the upregulation of HMGB1. Mature HMGB1 functioned as an effector to induce the expression of proinflammatory cytokines, causing inflammation and resulting in SIMD pathology. Using an in vitro AlphaScreen technology, we identified three small molecules that could inhibit the CUL4A-RBX1 interaction. Of them, PSSM0332 showed the strongest ability to inhibit the ubiquitination of NcoR1, and its administration in SIMD mice exhibited promising effects on decreasing the inflammatory response. Collectively, our results reveal that the CRL4A^DCAF8 E3 ligase is critical for the initiation of SIMD by regulating the expression of HMGB1 and proinflammatory cytokines. Our results suggest that PSSM0332 is a promising candidate to inhibit the inflammatory response in the pathogenesis of SIMD, which will provide a new option for the therapy of SIMD.

8 illus, 34 ref

043315   XU X, HU M, YING R, ZOU J, LIN L, CHENG H, ZHOU R (Wuhan Univ, Wuhan 430072, P. R. China, Email: hhcheng@whu.edu.cn) : RAB37 multiple alleles, transcription activation and evolution in mammals. Int J Biol Sci 2020, 16(15), 2964-73.

Detecting selection signatures in genomes that relates to transcription regulation has been challenges in genetic analysis. Here, we report a set of transcription factors EBF1, E2F1 and EGR2 for transcription activation of RAB37 promoter by a comparative analysis of promoter activities of RAB37 in humans, mice, and pigs. Two of the transcription factors bound to and co-regulated RAB37 promoter in each species. SNPs were further screened in pig RAB37 gene by population genomics in pig populations from both China and Europe. Three SNPs were identified in second CpG island upstream of core promoter of RAB37. These SNP variations led to at least 5 haplotypes, representing 5 multiple alleles of RAB37 in pig population. Distribution of these alleles in different genetic background of breeds showed a role of artificial selection for the variations of these multiple alleles. Of them, RAB37-c acquired the highest ability to activate gene expression in comparison with the other promoters, thus enhanced autophagy efficiently. These findings provide better understanding of transcription activation of RAB37 and artificial selection via RAB37 for autophagy regulation.

6 illus, 1 table, 25 ref

043314   XU L, WANG J, YUAN X, YANG S, XU X, LI K, HE Y, WEI L, ZHANG J, TIAN Y (Human Anatomy Dep, Wuhan Univ, Wuhan, Hubei 430071, P.R. China, Email: yihaotian@whu.edu.cn) : IU1 suppresses proliferation of cervical cancer cells through MDM2 degradation. Int J Biol Sci 2020, 16(15), 2951-63.

Previous studies have demonstrated that the antitumor potential of IU1 (a pharmacological compound), which was mediated by selective inhibition of proteasome-associated deubiquitinase ubiquitin-specific protease 14 (USP14). However, the underlying molecular mechanisms remain elusive. It has been well established that mdm2 (Murine double minute 2) gene was amplified and/or overexpressed in a variety of human neoplasms, including cervical cancer. Furthermore, MDM2 is critical to cervical cancer development and progression. Relatively studies have reported that USP15 and USP7 stabilized MDM2 protein levels by removing its ubiquitin chain. In the current study, we studied the cell proliferation status after IU1 treatment and the USP14-MDM2 protein interaction in cervical cancer cells. This study experimentally revealed that IU1 treatment reduced MDM2 protein expression in HeLa cervical cancer cells, along with the activation of autophagy-lysosomal protein degradation and promotion of ubiquitin-proteasome system (UPS) function, thereby blocked G0/G1 to S phase transition, decreased cell growth and triggered cell apoptosis. Thus, these results indicate that IU1 treatment simultaneously targets two major intracellular protein degradation systems, ubiquitin-proteasome and autophagy-lysosome systems, which leads to MDM2 degradation and contributes to the antitumor effect of IU1.

8 illus, 43 ref

043313   LIU G, GUO W, CHEN G, LI W, CUI Y, QIN J, PENG J (Cardiothoracic surgery Dep, The Affiliated Zhuzhou Hospital of Xiangya Medical Coll CSU, Zhuzhou 412000, China, Email: drjingpeng@163.com) : Lnc-MCEI mediated the chemosensitivity of esophageal squamous cell carcinoma via miR-6759-5p to competitively regulate IGF2. Int J Biol Sci 2020, 16(15), 2938-50.

Large amounts of long non-coding RNAs (lncRNAs) have been annotated whereas most of them have not been functionally characterized. Here we identified lncRNA ENST00000441932 as an oncogenic lncRNA in esophageal squamous cell carcinoma (ESCC) and named lnc-MCEI (lncRNA mediated the chemosensitivity of ESCC by regulating IGF2). What’s more, the effect of lnc-MCEI on the chemosensitivity of ESCC was further evaluated. Bioinformatics analysis demonstrated that lnc-MCEI was involved in the tumorigenesis of ESCC and lnc-MCEI levels were significantly increased in ESCC cells and tissues. Additionally, lnc-MCEI knockdown retarded cell proliferation, colony formation of ESCC cells, but induced cell apoptosis. Moreover, lnc-MCEI knockdown significantly improved the chemosensitivity of ESCC to cisplatin (DDP) both in vivo and in vitro. Further mechanisms disclosed that lnc-MCEI functioned as a competing endogenous RNA (ceRNA) via sponging miR-6759-5p and IGF2 was a target of miR-6759-5p. Meanwhile, we found that IGF2 suppressed chemosensitivity of ESCC cells via PI3K/AKT pathway. These data suggested that lnc-MCEI was an oncogenic lncRNA and lnc-MCEI knockdown enhanced chemosensitivity of ESCC cells to cisplatin by targeting miR-6759-5p /IGF2/PI3K/AKT axis.

8 illus, 38 ref

046603   RADHA P, UDHAYAVANI C, NAGARAJ R, SIVARANJANI K (Central Council for Research in Siddha, Salem District, Tamil Nadu, Email: radhasudar@rediffmail.com) : Ethno-gynaecological knowledge on medicinal plants among the rural communities of Tiruppur district, Tamil Nadu, India. Med Plants 2020, 12(4), 656-65.

The gynaecological disorder is one of the most common problem for most of the women in the recent years because of hormonal changes, fungal/bacterial infections, malnutrition and mental stress. The present work is a part of medicoethnobotanical survey conducted to document the medicinal knowledge of traditional practitioners among the rural and urban communities of Tiruppur District, Tamil Nadu, India. Frequent surveys are being conducted during the period of December 2017 - February 2020 to document the medicinal plants used for different gynaecological ailments. Information on folk/traditional medicine used to cure various gynaecological disorders has been documented using questionnaires and personal interviews with folk/traditional healers and housewives/elders of Tiruppur District. Demographic profile of the informants and local names of plant/animal/minerals used, mode of application and administration have documented. A total of 48 plant species, belonging to 47 genera spreading over 36 families were documented are found to be used in the treatment of various gynaecological disorders. It was observed that, the herbs are predominantly used for the gynaecological disorders. In addition to the medicinal plants, three minerals, two herbo-minerals and a marine animal product are also used. The documentation of the present work can be used for further research on bioprospecting on medicinal plants to find a novel drug discovery to cure various gynaecological diseases.

4 illus, 3 tables, 19 ref

041520   HU J, HU X, SHAN Z, WANG R (Anhui Sanlian Univ, Anhui Province, China, Email: erhu1999@126.com) : The efficacy of PBL model in pathology and pathophysiology teaching in China: A meta-analysis. Indian J Pharm Edu Res 2020, 54(3), 556-64.

To systematically evaluate the effect of PBL in Pathology and Pathophysiology teaching. CNKI, PubMed, WanFang Data, EMbase databases were electronically searched to collect randomized controlled trials (RCTs) of PBL model used in pathology and pathophysiology teaching in China from the database that has been constructed by September 20, 2019. Two researchers from the same research department independently screened and extracted literature materials for studying the evaluation bias risk and conducted a Meta-analysis using RevMan5.3 software. A total of 45 RCTs were enrolled, including 7,739 subjects. The Meta-analysis results indicate that PBL model in pathology and pathophysiology teaching is superior to traditional teaching model (LBL) in terms of final examination score [MD=6.68, 95 % CI(5.29,8.06), P<0.00001], case analysis score [MD=4.15, 95 % CI (2.88,5.42), P<0.00001], increased learning interest [RR=1.46, 95 %CI(1.28,1.66), P<0.00001], the ability to analyze and solve problems [RR=2.21, 95 %CI(1.49,3.27), P<0.00001] and teamwork ability [RR=1.7, 95 %CI(1.3,2.22), P<0.00001]. The research results shown that PBL model can improve the teaching effect of Pathology and Pathophysiology, which, however, needs to be further verified by more high quality researches due to the limitation of literature quality in this research.

7 illus, 2 tables, 49 ref

041519   DESAI S, TATKE P, GAITONDE V, GABHE S (SNDT Women’s Univ, Mumbai - 400 049, Maharashtra, Email: patatke@gmail.com) : Fast HPLC-DAD method for estimation of catechin for standardization of extracts and antiasthmatic polyherbal formulations containing Albizia lebbeck. Indian J Pharm Edu Res 2020, 54(3), 550-5.

Many marketed polyherbal formulations containing Albizia lebbeck are available for treatment of allergic disorders such as asthma and bronchitis. But these formulations are not standardized in terms of markers. The present work discusses development and validation of a new, simple and rapid HPLC-DAD method for quantification of phytomarker- catechin in methanol extracts for standardization of extracts and polyherbal formulations containing Albizia lebbeck HPLC analysis was carried out using C18 column (100 mm X 4.6 mm, 2.6 µm). 0.1 % Phosphoric acid and acetonitrile in gradient mode was used as mobile phase. The column oven was equilibrated at 28°C. The injection volume was 2 µl and detection was carried out at 279 nm. The developed HPLC method was found to be linear over the range of 3.0-60 µg/ml with correlation coefficient of 0.9998. The LOD and LOQ were found to be 1.0 µg/ml and 3.0 µg/ml, respectively. The recovery of the catechin was between 97.12 % -100.04 %. The low RSD values of intra and inter-day precision studies indicated good precision of the developed method. The method was found to be robust as small changes in method parameters did not significantly change peak symmetry and content of catechin. The developed method was sensitive, rapid and requires no special technique for sample preparation. The method was successfully applied for standardization of three bark extracts and marketed polyherbal formulations containing Albizia lebbeck.

2 illus, 4 tables, 15 ref

041518   GANAPA M S, MHATRE S S, RAUT S Y, PANDEY A, KALTHUR G, MUTALIK S (Pharmaceutics Dep, Manipal Coll of Pharmaceutical Sciences, Manipal - 576 104, Karnataka, Email: ss.mutalik@manipal.edu) : Novel oral drug delivery systems for steroids: Overview and recent updates. Indian J Pharm Edu Res 2020, 54(3), 541-9.

This review summarises the oral route of drug delivery of steroids. Oral drug delivery is widespread owing to its non-invasive nature which complements high patient compliance. This article summarises the problems associated with oral delivery of steroids including bioavailability. In this review, a brief description of the novel types of drug delivery systems of steroids such as microspheres, nanoparticles, solid lipid nanoparticles, multimatrix tablets, pellet technology, liposomes are presented. The review also presents the latest innovations and advances that have been achieved in recent years.

1 table, 39 ref

041517   BADHE R V, GODSE A, AHINKAR A (Pharmaceutical Quality Assurance Dep, Dr. D.Y. Patil Institute of Pharmaceutical Sciences and Research, Pune - 411 018, Maharashtra, Email: ravindra.badhe@dypvp.edu.in) : Biomaterials in 3D printing: A special emphasis on nanocellulose. Indian J Pharm Edu Res 2020, 54(3), 526-40.

The main objective of this review is to highlight nanocellulosic materials in 3D bioprinting. Three-dimensional (3D) bioprinting is on the verge of fabricating the artificial organ and living tissues. For the target construction the process of this 3D bioprinting involves layer-by-layer deposition of suitable biomaterials using predesigned data made by using Computer Aided Design (CAD) as an outline. However, only a handful of biomaterials are able to fulfil the considerable requirements for suitable bioink formulation, which is a critical component of efficient 3D bioprinting. Cellulose, a naturally occurring polysaccharide, is clearly the most commonly employed material in current bioinks. Here, in this review we discuss the advantages, reasons, applications, disadvantages of the use of cellulosic bioink in 3D bioprinting by summarizing the most recent studies that used cellulose for printing vascular tissue, bone and cartilage. In addition, other breakthroughs in the use of cellulose in bioprinting are discussed, including strategies to improve its structural and degradation characteristics. In this review, we organize the available literature in order to inspire and accelerate novel cellulose-based bioink formulations with enhanced properties for future applications in basic research, drug screening and regenerative medicine.

5 illus, 3 tables, 92 ref

043312   SONG Y, WU X, XU Y, ZHU J, LI J, ZOU Z, CHEN L, ZHANG B, HUA C, RUI H, et al. (Dermatology and Venereology Dep, Sir Run Run Shaw Hospital Zhejiang Univ School of Medicine, Zhejiang Province, China, 310016, Email: wqq@zju.edu.cn) : HPV E7 inhibits cell pyroptosis by promoting TRIM21- mediated degradation and ubiquitination of the IFI16 inflammasome. Int J Biol Sci 2020, 16(15), 2924-37.

Human papillomavirus (HPV) is a DNA virus that causes sexually transmitted infections. The HPV oncoprotein E7 plays a critical role in the regulation of host immunity to promote the immune escape of HPV and the occurrence of cervical cancer or genital warts. Pyroptosis, a highly inflammatory form of programmed cell death, can be induced by inflammasomes and acts as a defense against pathogenic infection. However, whether HPV E7 can regulate cell pyroptosis to evade immune surveillance has not been determined. In this study, we found that HPV E7 could inhibit cell pyroptosis induced by transfection with dsDNA. The activation of the inflammasome, and the production of IL-18 and IL-1β were also restrained by HPV E7. Mass spectrometry and immunoprecipitation showed that HPV E7 interacted with IFI16 and TRIM21. We also discovered that HPV E7 recruited the E3 ligase TRIM21 to ubiquitinate and degrade the IFI16 inflammasome, leading to the inhibition of cell pyroptosis and self-escape from immune surveillance. Thus, our study reveals an important immune escape mechanism in HPV infection and may provide targets for the development of a novel immunotherapeutic strategy to effectively restore antiviral immunity.

8 illus, 59 ref

041516   YU P, WILSON B J, BOBBINS A C, SRINIVAS S C, YIN G (Jinan Univ, Guangdong Province, China, Email: pennypeiyu@163.com) : Pharm D program: Is it necessary in BRICS?. Indian J Pharm Edu Res 2020, 54(3), 517-25.

The various trajectories of the development of the Doctor of Pharmacy (Pharm D) programs in BRICS countries were reviewed individually, showing the progress of Pharm D programs in each context. Evidence from developed countries indicates that Pharm D programs have contributed towards the positive development of public health and a country’s local health requirements, with well-trained clinical pharmacists having a key role in the health and economic development of these emerging powers. The health systems of BRICS countries are heavily challenged with a dual burden of disease, weak health infrastructure and public health initiatives. Thus, not only will the development of Pharm D programs in BRICS countries strengthen pharmacy education contributing to a pharmacy workforce more readily able to target health-related problems, demographic and epidemiological shifts; but will also better equip public health systems with more patient-centered pharmaceutical care necessary for evidence-based, individualized and cost-effective healthcare.

1 table, 47 ref

043311   WANG C, TIAN Q, ZHAO P, ZIONG M, LATKIN C A, GAN Y, HALL B J, YANG B (Southern Medical Univ, Guangdong, China, Email: yangbin101@hotmail.com) : Disease knowledge and attitudes during the COVID-19 epidemic among international migrants in China: A national cross-sectional study. Int J Biol Sci 2020, 16(15), 2895-905.

There are more than 258 million international migrants worldwide and the majority reside in countries with ongoing novel coronavirus disease 2019 (COVID-19) epidemic outbreaks. International migrants may not receive adequate and timely disease information during epidemics, increasing vulnerability to disease transmission. This is one of very limited studies focusing on international migrants’ COVID-19 prevention knowledge and attitudes during the epidemic. A national cross-sectional online survey was conducted across 100 cities and 26 regions in China from February 17 and March 1, 2020. The sample included 1,426 international migrants representing 77 countries and 6 continents. Knowledge was defined as the number of correct responses to questions about COVID-19. Attitudes included worries, expectations, and general preparedness. Multivariable ordinal logistic regressions evaluated correlates of knowledge and attitudes including information channels and preferences, and trust in Chinese institutions and groups.  Just half of the sample, 730/1426 (51.2 %) had a good level of knowledge and 656/1426 (46.0 %) had a positive attitude towards the COVID-19 epidemic. Knowledge was associated with receiving information through social media (aOR: 2.0, 95 %CI: 1.2-3.2), the Internet (aOR: 1.4, 95 %CI: 1.2-1.8), the community (aOR: 1.5, 95 %CI: 1.2-1.8), and encountering language barriers when receiving medical services (aOR: 0.8, 95 %CI: 0.7-1.0). Positive attitude was associated with the level of trust in various Chinese institutions and groups. Roughly half of the sample reported inadequate knowledge and poor attitudes toward prevention and control of COVID-19. Tailored public health campaigns are needed to ensure that international migrants possess adequate knowledge to protect their health during future epidemics and disasters.

5 tables, 20 ref

041515   AGA Q A A K, NAIR A B, HOURANI W, PARASURAMAN S, AHMED KK M, SREEHARSHA Nq (Pharmaceutical Sciences Dep, Philadelphia Univ, Amman, Jordan, Email: qutaiba975@gmail.com) : Therapeutic potential of chymase inhibitors in cardiovascular diseases: An overview. Indian J Pharm Edu Res 2020, 54(3), 509-16.

Chymase belongs to the family of serine proteases and is mainly warehoused in a heparin proteoglycan macromolecular complex within the mast cells. Extensive studies have been carried out in the last few decades to assess the role of chymase in human diseases. Recent studies have shown the significance of chymase in blood pressure regulation owing to its efficient angiotensin II forming activity. Angiotensin II-generation routes that are associated with human cardiovascular diseases have pathophysiological importance which is still argumentative. Chymase inhibitors play a distinctive role in regulating the renin-angiotensin system as compared to the inhibitors of angiotensin-converting enzyme and type 1 angiotensin II receptor. Therefore, this mechanism may have a role in medical applications of chymase inhibitors by inactivation of the local renin-angiotensin system to avoid cardiovascular diseases. This review highlights the significant role of chymase inhibitors as a potential approach for the management of cardiovascular diseases such as atherosclerosis, hypertension, vascular proliferation, myocardial infarction and heart failure.

5 illus, 59 ref

043310   YAO Y, CUI L, YE J, YANG G, LU G, FANG X, ZENG Z, ZHOU J (Respiratory Medicine Dep, Zhejiang Univ, Hangzhou, China, Email: zjyhz@zju.edu.cn) : Dioscin facilitates ROS-induced apoptosis via the p38-MAPK/HSP27-mediated pathways in lung squamous cell carcinoma. Int J Biol Sci 2020, 16(15), 2883-94.

Lung squamous cell carcinoma (SCC) is one of the deadliest cancers both in China and worldwide. To date, the efficacy of lung SCC treatments is limited. Recent studies have elucidated the powerful anti-tumour role of dioscin in different human cancers. Here, our study aims to investigate the effect of dioscin on lung SCC and its underlying mechanism. First, we found that dioscin not only inhibited cell proliferation and cell migration and induced cell apoptosis in lung SCC cells but also suppressed tumour growth in tumour-bearing mice. Furthermore, we noted that the accumulation of intracellular reactive oxygen species (ROS) was triggered by dioscin in lung SCC cells, leading to the phosphorylation of HSP27 through p38-MAPK and consequent cell apoptosis. The activation of p38-MAPK/HSP27 induced by the p38-MAPK activator Anisomycin enhanced the apoptosis of lung SCC cells, while the ROS inhibitor N-acetyl-L-cysteine (NAC) and the p38-MAPK inhibitor SB203580 both attenuated dioscin-mediated cell apoptosis. Moreover, NAC suppressed the activation of p38-MAPK/HSP27 that induced by dioscin. In conclusion, these results confirm that dioscin facilitates ROS-induced apoptosis via the p38-MAPK/HSP27-mediated pathway in lung SCC.

7 illus, 38 ref

041514   SAQIB A, PATTAR S, KARIGAR C S, SEKHAR S (Mysore Univ, Mysuru - 570 006, Karnataka, Email: shailasree@ioe.uni-mysore.ac.in) : Caspase activators: Phytochemicals with apoptotic properties targeting cancer, a health care strategy to combat this disease. Indian J Pharm Edu Res 2020, 54(3), 496-508.

Caspases, a family of cysteine-aspartic proteases have a pivotal role in apoptotic pathways. Their down-regulation is reported to induce inappropriate cell survival and enhanced carcinogenic potential. Screening of phytochemicals with a capacity to activate caspases enhancing apoptotic capacity has been proven to be effective anticancer agents. This review consolidates data on phtochemicals traditionally used to treat cancerous conditions. The scientific validation of caspaseactivated apoptosis for this traditional application has been compiled. nternet assisted scientific literature was collected from Google, Google Scholar, Research Gate and NCI, restricted to publications from 1997 to 2019. Search terms ‘caspases and cancer’, ‘assay of caspases’, ‘traditionally used medicinal plants’, ‘Kani tribes’, ‘plant extracts activating caspase’, ‘cytotoxicity assay’, ‘docking phytochemicals to caspase’, ‘technological advancement for anticancer therapy’, ‘clinical studies of plant extracts and phytochemicals’ and ‘herbal drugs approved by FDA’ was included. The compilation revealed significance of multiple experimental strategies, traditional research laboratory practices and advanced in silico molecular docking techniques in anticancer therapy. Technological advancement such as MALDI-TOF assisted phytochemical mediated protein target identification and designing promoter for caspases activation and synthesizing functionalized nano carriers for clinical studies has been included for identification of hit molecule and lead optimization. Eugenol and berberine were identified as phytochemicals with potential drug characteristics by both in silico and in vivo studies. The phytochemicals from important Kani tribal medicinal plants via in silico docking and in vivo studies identified could be explored at clinical trials.

4 illus, 5 tables, 86 ref

043309   ZHANG F, WANG C, JIANG Y, HUANG K, LIU F, DU M, LUO X, HUANG D, HUANG K (Cardiovascular Diseases Dep, Huazhong Univ of Science and Technology, Hubei, China, 430074, Email: xiaoyou_321@foxmail.com) : Yin and yang regulation of liver x receptor α signaling control of cholesterol metabolism by poly(adp-ribose) polymerase 1. Int J Biol Sci 2020, 16(15), 2868-82.

Liver X receptor α (LXRα) controls a set of key genes involved in cholesterol metabolism. However, the molecular mechanism of this regulation remains unknown. The regulatory role of poly(ADP-ribose) polymerase 1 (PARP1) in cholesterol metabolism in the liver was examined. Activation of PARP1 in the liver suppressed LXRα sensing and prevented upregulation of genes involved in HCD-induced cholesterol disposal. Mechanistically, LXRα was poly(ADP-ribosyl)ated by activated PARP1, which decreased DNA binding capacity of LXRα, thus preventing its recruitment to the target promoter. Intriguingly, we found that unactivated PARP1 was indispensable for LXRα transactivation and target expression. Further exploration identified unactivated PARP1 as an essential component of the LXRα-promoter complex. Taken together, the results indicate that activated PARP1 suppresses LXRα activation through poly(ADP-ribosyl)ation, while unactivated PARP1 promotes LXRα activation through physical interaction. PARP1 is a pivotal regulator of LXRα signaling and cholesterol metabolism in the liver.

7 illus, 1 table, 39 ref

041513   DEVI S, SINGH K (Medical Lab Technology Dep, Galgotias Univ, Greater Noida - 201 310, Uttar Pradesh, Email: kamaldeepbilkhu53@gmail.com) : Is Covid-19 sibling of SARS AND MERS? a review on: Novel Covid-19. Indian J Pharm Edu Res 2020, 54(3), 491-5.

Pandemic times: Contemporary public health catastrophe terrifying the world over with the rapidly spreading of 2019-nCoV or SARS-CoV-2. Coronaviruses have pass over the species barriers to cause fatal disease in human since the 2002 and 2012; Severe Acute Respiratory Syndrome and Middle East Respiratory Syndrome respectively. It was suggested that SARS-CoV and SARS-CoV-2 are firmly related to each and hypothesized it originated in animal (bats) and was transmitted to humans. Though Novel Covid-19’s intermediary animal is not clear. As a matter of fact, transmission of Covid-19 is direct contact with infected people or inhalation of droplets thus; escalate its spread. World Health Organization stated the novel SARS-CoV-2 is one of the epidemic disease on January 30, 2020 and on March 11, 2020 declared a pandemic the rapidly spreading SARS-CoV-2- a globe first for corona virus. Additionally, the incubation phase is from 2 to 14 days. Usually the sign and symptoms are high temperature, dry cough, tiredness, sore throat, difficulty breathing (severe cases) AND malaise among others. The diagnosis can be done with Real Time Polymerase Chain Reaction test to suspected SARS-CoV-2 infection along with chest X-ray. Till time, no medicine or vaccines are prepared that can approved against SARS-CoV-2 or any human-infecting corona viruses. But some researchers have confirmed to curtail its effects by administrating antiviral drugs such as Oseltamivir, Ribavirin, Ganciclovir, Lopinavir AND Ritonavir.

1 illus, 1 table, 30 ref

041512   PATIL R Y, MORE H N (Pharmacology Dep, D.S.T.S Mandal’s Coll of Pharmacy, Solapur - 413 004, Maharashtra, Email: rypatilsir@gmail.com) : Antioxidants with multivitamin and mineral supplementation attenuates chemotherapy or radiotherapy-induced oxidative stress in cancer patients. Indian J Pharm Edu Res 2020, 54(2), 484-90.

The present study was planned to evaluate the combined effects of antioxidant multivitamin and minerals (AMM) in reducing cancer and cancer therapy-induced oxidative stress. In addition cell viability of human malignant melanoma and normal mouse fibroblast cell lines after treatment with different concentrations of AMM and methotrexate were also estimated. Amongst the 120 cancer patients recruited for the study, 60 patients each were treated with radiotherapy and chemotherapy. In both radiotherapy and chemotherapy, 30 patients each were supplemented with AMM for 30 days. Thirty healthy human volunteers were recruited for comparison. On day 31, blood samples were collected for estimation of oxidative stress markers (MDA and nitrite), endogenous (SOD and GPx) and exogenous (Vitamin C and Vitamin E) antioxidants and essential trace elements (zinc, copper and selenium). In addition, cell viability of human malignant melanoma and normal mouse fibroblast cell lines were estimated after treatment with different concentrations of AMM and methotrexate. Supplementation of AMM during cancer therapy minimized the burden of free reactive radicals and restored the endogenous and exogenous antioxidants and essential trace element levels. In addition, AMM has no cytotoxic effect on normal mouse fibroblast cell lines. But AMM imparts suppression of cell proliferation in human malignant melanoma. Supplementation of antioxidants with multivitamin and mineral during cancer therapy is found to be beneficial as they minimize the burden of free reactive radicals and restores the endogenous and exogenous antioxidants and essential trace element levels.

4 illus, 2 tables, 18 ref

043308   HU C, YU M, LI C, WANG Y, LI X, ULRICH B, SU R, DONG L, WENG H, HUANG H, et al. (Hematology Dep, First Affiliated Hospital of Zhejiang Univ, Zhejiang 310003, China, Email: jiej0503@zju.edu.cn) : Mir-550-1 functions as a tumor suppressor in acute myeloid leukemia via the hippo signaling pathway. Int J Biol Sci 2020, 16(15), 2853-67.

MicroRNAs (miRNAs) and N6-methyladenosine (m6A) are known to serve as key regulators of acute myeloid leukemia (AML). Our previous microarray analysis indicated miR-550-1 was significantly downregulated in AML. The specific biological roles of miR-550-1 and its indirect interactions and regulation of m6A in AML, however, remain poorly understood. At the present study, we found that miR-550-1 was significantly down-regulated in primary AML samples from human patients, likely owing to hypermethylation of the associated CpG islands. When miR-550-1 expression was induced, it impaired AML cell proliferation both in vitro and in vivo, thus suppressing tumor development. When ectopically expressed, miR-550-1 drove the G0/1 cell cycle phase arrest, differentiation, and apoptotic death of affected cells. We confirmed mechanistically that WW-domain containing transcription regulator-1 (WWTR1) gene was a downstream target of miR-550-1. Moreover, we also identified Wilms tumor 1-associated protein (WTAP), a vital component of the m6A methyltransferase complex, as a target of miR-550-1. These data indicated that miR-550-1 might mediate a decrease in m6A levels via targeting WTAP, which led to a further reduction in WWTR1 stability. Using gain- and loss-of-function approaches, we were able to determine that miR-550-1 disrupted the proliferation and tumorigenesis of AML cells at least in part via the direct targeting of WWTR1. Taken together, our results provide direct evidence that miR-550-1 acts as a tumor suppressor in the context of AML pathogenesis, suggesting that efforts to bolster miR-550-1 expression in AML patients may thus be a viable clinical strategy to improve patient outcomes.

7 illus, 2 tables, 60 ref

043307   DONG X, FENG M, YANG H, LIU H, GUO H, GAO X, LIU Y, LIU R, ZHANG N, CHEN R, et al. (Tianjin Univ, Tianjin, 300072, P.R. China, Email: rbchen@tju.edu.cn) : Rictor promotes cell migration and actin polymerization through regulating ABLIM1 phosphorylation in hepatocellular carcinoma. Int J Biol Sci 2020, 16(15), 2835-52.

As one of the most ominous malignancies, hepatocellular carcinoma (HCC) is frequently diagnosed at an advanced stage, owing to its aggressive invasion and metastatic spread. Emerging evidence has demonstrated that Rictor, as a unique component of the mTORC2, plays a role in cell migration, as it is dysregulated in various cancers, including HCC. However, the underlying molecular mechanism has not been well-characterized. Here, evaluation on a tissue-array panel and bioinformatics analysis revealed that Rictor is highly expressed in HCC tissues. Moreover, increased Rictor expression predicts poor survival of HCC patients. Rictor knockdown significantly suppressed cell migration and actin polymerization, thereby leading to decreased nuclear accumulation of MKL1 and subsequent inactivation of SRF/MKL1-dependent gene transcription, i.e. Arp3 and c-Fos. Mechanistically, we identified ABLIM1 as a previously unknown phosphorylation target of Rictor. Rictor interacts with ABLIM1 and regulates its serine phosphorylation in HCC cells. We generated ABLIM1 knockout cell lines of HCC, in which dominant negative mutations of Ser 214 and Ser 431 residues inhibited the ABLIM1-mediated actin polymerization and the MKL1 signaling pathway. Overall, ABLIM1 phosphorylation induced by Rictor plays an important role in controlling actin polymerization in HCC cells.

8 illus, 44 ref

043306   HE Z, CHEN J, PAN K, YUE Y, CHEUNG T, YUAN Y, DU N, ZHAO Y, FENG Y, DHOU D, et al. (Electronic Science and Technology of China Univ, Chengdu, 610054, PR China, Email: 2357622298@qq.com) : The development of the ‘covid-19 psychological resilience model’ and its efficacy during the covid-19 pandemic in China. Int J Biol Sci 2020, 16(15), 2828-34.

During the novel coronavirus disease 2019 (COVID-19) outbreak, traditional face-to-face psychological interventions have been suspended due to high risks of rapid transmission. Developing an effective online model of psychological intervention is deemed necessary to deal with the mental health challenges brought up by this disease. An integrated psychological intervention model coined ‘COVID-19 Psychological Resilience Model’ was developed in Chengdu, China including live media, 24-hour hotline consultations, online video intervention and on-site crisis intervention sessions to provide services to those in need. A total of 45 episodes of live media programs on COVID-19 outbreak-related psychological problems were broadcasted with over 10 million views. A total of 4,236 hotline consultations were completed. More than 50 % of the clients had positive feedback about the hotline consultations. A total of 223 cases received online video intervention, of which 84.97 % were redirected from the hotline consultation and 15.03 % from COVID-19-designated hospital and community-based observation spots. Seventy one-on-one psychological interventions were conducted with 39 COVID-19 patients, and one-third were treated with medication. Additionally, 5 training sessions were conducted to 98 frontline medical staff. This ‘COVID-19 Psychological Resilience Model’ is proven effective to the general population during the COVID-19 pandemic. We have greatly improved the overall mental health of our target population during the COVID-19 pandemic. This model could provide valuable experiences and serve as a reference guide for other countries to offer effective psychological intervention, and reduce detrimental negative mental health outcomes in public health emergency.

4 illus, 1 table, 24 ref

043305   ZHANG D, LIU J, XIE T, JIANG Q, DING L, ZHU J, YE Q (Medical Molecular Biology Dep, Beijing Institute of Biotechnology, Beijing 100850, China, Email: yeqn66@yahoo.com) : Oleate acid-stimulated HMMR expression by CEBPα is associated with nonalcoholic steatohepatitis and hepatocellular carcinoma. Int J Biol Sci 2020, 16(15), 2812-27.

Non-alcoholic steatohepatitis (NASH) is a type of nonalcoholic fatty liver disease and has become a major risk factor for hepatocellular carcinoma (HCC). However, the underlying pathophysiological mechanisms are still elusive. Here, we identify hyaluronan-mediated motility receptor (HMMR) as a critical gene associated with NASH/HCC by combination of bioinformatic analysis and functional experiments. Analysis of differentially expressed genes (DEGs) between normal controls and NASH/HCC identified 5 hub genes (HMMR, UBE2T, TYMS, PTTG1 and GINS2). Based on the common DEGs, analyses of univariate and multivariate Cox regression and the area under the curve (AUC) value of the receiver operating characteristic (ROC) indicate that HMMR is the most significant gene associated with NASH/HCC among five hub genes. Oleate acid (OA), one of fatty acids that induce cellular adipogenesis, stimulates HMMR expression via CCAAT/enhancer-binding protein α (CEBPα). CEBPα increases the expression of HMMR through binding to its promoter. HMMR promotes HCC cell proliferation in vitro via activation of G1/S and G2/M checkpoint transitions, concomitant with a marked increase of the positive cell cycle regulators, including cyclin D1, cyclin E, and cyclin B1. Knockdown of HMMR suppresses HCC tumor growth in nude mice. Our study identifies an important role of HMMR in NASH/HCC, and suggests that HMMR may be a useful target for therapy and prognostic prediction of NASH/HCC patients.

7 illus, 75 ref

043304   HE M, MA Y, WANG R, ZHANG J, JING L, LI P A (Pharmaceutical Sciences Dep, North Carolina Central Univ, Durham, NC 27707, USA, Email: pli@nccu.edu) : Deletion of mitochondrial uncoupling protein 2 exacerbates mitochondrial damage in mice subjected to cerebral ischemia and reperfusion injury under both normo- and hyperglycemic conditions. Int J Biol Sci 2020, 16(15), 2788-802.

Deletion of mitochondrial uncoupling protein 2 (UCP2) has been shown to aggravate ischemic damage in the brain. However, the underlying mechanisms are not fully understood. The objective of this study is to explore the impact of homozygous UCP2 deletion (UCP2^-/- ) on mitochondrial fission and fusion dynamic balance in ischemic mice under normo- and hyperglycemic conditions. UCP2^-/- and wildtype mice were subjected to a 60 min middle cerebral artery occlusion (MCAO) and allowed reperfusion for 6h, 24h and 72h. Our results demonstrated that deletion of UCP2 enlarged infarct volumes and increased numbers of cell death in both normo- and hyperglycemic ischemic mice compared with their wildtype counterparts subjected to the same duration of ischemia and reperfusion. The detrimental effects of UCP deletion were associated with increased ROS production, elevated mitochondrial fission markers Drp1 and Fis1 and suppressed fusion markers Opa1 and Mfn2 in UCP2^-/- mice. Electron microscopic study demonstrated a marked mitochondrial swolling after 6h of reperfusion in UCP2^-/- mice, contrasting to a mild mitochondrial swolling in wildtype ischemic animals. It is concluded that the exacerbating effects of UCP2^-/- on ischemic outcome in both normo- and hyperglycemic animals are associated with increased ROS production, disturbed mitochondrial dynamic balance towards fission and early damage to mitochondrial ultrastructure.

9 illus, 1 table, 53 ref

043303   YAN F, WANG R, LI S, ZHAO X, JIANG Y, LIU L, FANG J, ZHEN X, LAZAROVICI P, ZHENG W (Macau Univ, 999078, P. R China, Email: wenhuazheng@um.edu.mo) : FoxO3a suppresses neuropeptide W expression in neuronal cells and in rat hypothalamus and its implication in hypothalamic-pituitary-adrenal (HPA) axis. Int J Biol Sci 2020, 16(15), 2775-87.

FoxO3a, a forkhead family member of transcription factors, is involved in the regulation of cell metabolism, proliferation, differentiation and apoptosis. However, whether FoxO3a participates in the regulation of glucocorticoids induced-hypothalamic-pituitary-adrenal (HPA) dysfunction is still unknown. Our present results indicate that dexamethasone(DEX) increased FoxO3a expression in PC12 and hypothalamic neuronal cultures in correlation to reduced expression of NPW, a process that could be blocked by GR2 antagonist. DEX restrained the phosphorylation of Akt and FoxO3a, but not ERK1/2 phosphorylation, resulting with FoxO3a nuclear localization. Overexpression of FoxO3a inhibited NPW expression, while FoxO3a knockdown by siRNA had the opposite effect. The regulatory region of NPW promoter contains multiple FoxO3a binding sites, and FoxO3a bonding to these sites inhibited its transcriptional activity. In a rat model, chronic administration of corticosterone reduced animals’ body weight and sucrose consumption and caused stress- depression like behavior. Corticosterone treatment induced a marked increase in FoxO3a levels, while decreased the expression of NPW protein in the hypothalamus. Immunofluorescent double labeling demonstrated that FoxO3a and NPW were collocated in the hypothalamus. Taken together, these data indicate that NPW is a new direct downstream target gene of FoxO3a. FoxO3a suppressed the transcription of NPW and modulated glucocorticoids-induced HPA dysfunction by directly regulating the expression of NPW. Thus, present findings suggest that FoxO3a and NPW may be potential therapeutic targets for endocrine and psychiatric disorders.

7 illus, 38 ref

041509   MAHAJAN M R, PATIL D D (Quality Assurance Dep, H. R. Patel Institute of Pharmaceutical Education and Research, Dhule - 425 405, Maharashtra, Email: dipakpatil888@gmail.com) : Robust analytical method for iron estimation by experimental design approach. Indian J Pharm Edu Res 2020, 54(2), 456-64.

To perform Iron estimation by UV-Visible spectroscopy using an Experimental design approach. The robust analytical method was developed for the estimation of iron (III) using 1, 10-phenanthroline reagent. The analytical method is an exploration of the chemical reaction of iron with 1, 10- phenanthroline reagent to form a colored complex which was measured in the UV-Visible region at 509 nm. To monitor the effect of diverse factors like the concentration of reagent (A), volume of reagent (B), pH (C) and time (D) on the formation of iron 1, 10 – phenanthroline complex the full factorial design (two-level) was used. From the Pareto chart, Normal plot and half normal plot, it was studied that a combination of all factors was initiate to be significant. Then significant variables are optimized by response surface methodology (RSM) via Box-Behnken design. The evaluation of design was performed to study the effect on the selected response by quadratic effects and main interaction effects. The contour plot and surface plot used for the determined response of the selected factors for their optimum value. The prime reaction state, Beer’ s law were obeyed in 2.0-10.0 µg/ml concentration range with a correlation coefficient of 0.998. The method was successfully applied for the estimation of iron in iron sucrose injection. The optimized method was used for the quantitative analysis of iron sucrose injection.

12 illus, 6 tables, 28 ref

043302   WANG H, SHI X, QIU M, LV S, ZHENG H, NIU B, LIU H (Henan Univ, Henan, 475000, China, Email: whg197167@vip.henu.edu.cn) : Hydrogen sulfide plays an important role by influencing NLRP3 inflammasome. Int J Biol Sci 2020, 16(14), 2752-60.

Inflammasome is a complex composed of several proteins and an important part of the natural immune system. Nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome is composed of NLRP3, apoptosis associated speck like protein (ASC) and pro-caspase-1. It plays an important role in many diseases. Hydrogen sulfide (H2S) is an important signaling molecule that regulates many physiological and pathological processes. Recent studies indicated that H2S played anti-inflammatory and pro-inflammatory roles in many diseases through influencing NLRP3 inflammasome, but its mechanism was not fully understood. This article reviewed the progress about the effects of H2S on NLRP3 inflammasome and its mechanisms involved in recent years to provide theoretical basis for in-depth study

4 illus, 102 ref

041507   RAO T M, REDDY C B, BABU P S (Pharmaceutical Sciences Dep, Jawaharlal Nehru Technological Univ, Kakinada - 533 003, Andhra Pradesh, Email: mohanjntuk.sch@gmail.com) : LC-MS/MS determination and pharmacokinetics study of apremilast after oral administration in rabbits. Indian J Pharm Edu Res 2020, 54(2), 440-7.

A selective and reproducible method has been optimised for evaluation of Apremilast in rabbit biological matrices by UPLC-ESI-MS/MS. The analytical technique was implemented to quantify the apremilast in rabbit plasma samples with Apremilast-D5 as deuterated internal standard. The chromatographic separation tuned with 10mM Ammonium Acetate Buffer (pH: 4.0): Methanol: Acetonitrile, (20:40:40 %, v/v/v) using the CORTECS C₁₈, 2.7 µ.m, 4.6 m.m X 150 m.m analytical column with analysis time four minutes. The flow of mobile phase through column is 0.5 m.L/min. The mass spectrometric ions of Apremilast and Apremilast-D5 obtained were m/z 461.5→257.1 and 466.5→257.1. The curve indicates correlation coefficient (r²) was superior than 0.998 with linear range of 0.03-48.0 n.g/m.L. The developed method was tuned to apply efficaciously for analyzing the pharmacokinetic parameters of Apremilast in rabbit plasma samples. An accurate and reproducible novel method was fabricated for estimation of Apremilast in rabbit biological matrices by UPLC-ESI-MS/MS will be used for regular analysis and appropriate for therapeutic drug monitoring.

6 illus, 4 tables, 37 ref

043301   ZHUGE Y, ZHANG J, QIAN F, WEN Z, WEN Z, NIU C, XU K, JI H, RONG X, CHU M, JIA C (Hospital of Wenzhou Medical Univ, Wenzhou 325027, China, Email: chmping@hotmail.com) : Role of smooth muscle cells in Cardiovascular Disease. Int J Biol Sci 2020, 16(14), 2741-51.

Normally, smooth muscle cells (SMCs) are localized in the tunica media of the vasculature, where they take responsibility for vascular contraction and extracellular matrix (ECM) generation. SMCs also play a significant role in obedience and elastic rebound of the artery in response to the haemodynamic condition. However, under pathological or stressed conditions, phenotype switching from contractile to synthetic state or other cell types will occur in SMCs to positively or negatively contribute to disease progression. Various studies demonstrated that functional changes of SMCs are implicated in several cardiovascular diseases. In this review, we present the function of vascular SMCs (VSMCs) and the involved molecular mechanisms about phenotype switching, and summarize the roles of SMCs in atherosclerosis, hypertension, arterial aneurysms and myocardial infarction, hoping to obtain potential therapeutic targets against cardiovascular disease in the clinical practices.

4 illus, 98 ref

043300   ZHANG Y, QIAN H, WU B, YOU S, WU S, LU S, WANG P, CAO L, ZHANG N, SUN Y (Cardiology Dep, First Hospital of China Medical Univ, Shenyang, Liaoning, P.R. China, Email: njzhang@cmu.edu.cn) : E3 Ubiquitin ligase NEDD4 family‑regulatory network in cardiovascular disease. Int J Biol Sci 2020, 16(14), 2727-40.

Protein ubiquitination represents a critical modification occurring after translation. E3 ligase catalyzes the covalent binding of ubiquitin to the protein substrate, which could be degraded. Ubiquitination as an important protein post-translational modification is closely related to cardiovascular disease. The NEDD4 family, belonging to HECT class of E3 ubiquitin ligases can recognize different substrate proteins, including PTEN, ENaC, Nav1.5, SMAD2, PARP1, Septin4, ALK1, SERCA2a, TGFβR3 and so on, via the WW domain to catalyze ubiquitination, thus participating in multiple cardiovascular-related disease such as hypertension, arrhythmia, myocardial infarction, heart failure, cardiotoxicity, cardiac hypertrophy, myocardial fibrosis, cardiac remodeling, atherosclerosis, pulmonary hypertension and heart valve disease. However, there is currently no review comprehensively clarifying the important role of NEDD4 family proteins in the cardiovascular system. Therefore, the present review summarized recent studies about NEDD4 family members in cardiovascular disease, providing novel insights into the prevention and treatment of cardiovascular disease. In addition, assessing transgenic animals and performing gene silencing would further identify the ubiquitination targets of NEDD4. NEDD4 quantification in clinical samples would also constitute an important method for determining NEDD4 significance in cardiovascular disease.

3illus, 1 table, 177 ref

043298   ZHANG Q, DONG J, YU Z (Affiliated Cancer Hospital and Institute of Guangzhou Medical Univ, Heng Zhi Gang Road 78, Guangzhou, 510095, Email: gjw94@nefu.edu.cn) : Pleiotropic use of statins as non-lipid-lowering drugs. Int J Biol Sci 2020, 16(14), 2704-11.

Statins, known as HMG-CoA reductase (HMGCR) inhibitors, have primarily been utilized for metabolic and angiographic medical applications because of their cholesterol-lowering effects. Similar to other drugs, statins may also induce a series of potential side effects. Statins inhibit the HMGCR (rate-limiting enzyme) activity in early stages of mevalonate pathway and then indirectly affect a number of intermediate products, including non-sterol isoprenoids (coenzyme Q10, dolichol etc.), which can result in impaired functions of body organs. Recently, scores of studies have uncovered additional functional mechanisms of statins in other diseases, such as diabetes mellitus, nervous system diseases, coronary heart disease, inflammation and cancers. This review aims to summarize the positive and adverse mechanisms of statin therapy. Statin care should be taken in the treatment of many diseases including cancers. Since the underlying mechanisms are not fully elucidated, future studies should spend more time and efforts on basic research to explore the mechanisms of statins.

5 illus, 72 ref

043297   WANG S, DENG Z, MA Y, JIN J, QI F, LI S, LIU C, LYU F-J, ZHENG Q (Orthopedics Dep, South China Univ, Yuexiu District, Guangzhou, 510080, China, Email: zhengqiujian@gdph.org.cn) : The role of autophagy and mitophagy in bone metabolic disorders. Int J Biol Sci 2020, 16(14), 2675-91.

Bone metabolic disorders include osteolysis, osteoporosis, osteoarthritis and rheumatoid arthritis. Osteoblasts and osteoclasts are two major types of cells in bone constituting homeostasis. The imbalance between bone formation by osteoblasts and bone resorption by osteoclasts has been shown to have a direct contribution to the onset of these diseases. Recent evidence indicates that autophagy and mitophagy, the selective autophagy of mitochondria, may play a vital role in regulating the proliferation, differentiation and function of osteoblasts and osteoclasts. Several signaling pathways, including PINK1/Parkin, SIRT1, MAPK8/FOXO3, Beclin-1/BECN1, p62/SQSTM1, and mTOR pathways, have been implied in the regulation of autophagy and mitophagy in these cells. Here we review the current progress about the regulation of autophagy and mitophagy in osteoblasts and osteoclasts in these bone metabolic disorders, as well as the molecular signaling activated or deactivated during this process. Together, we hope to draw attention to the role of autophagy and mitophagy in bone metabolic disorders, and their potential as a new target for the treatment of bone metabolic diseases and the requirements of further mechanism studies.

6 illus, 1 table, 159 ref

043296   HE B, ZHAO Z, CAI Q, ZHANG Y, ZHANG P, SHI S, XIE H, PENG X, YIN W, TAO Y, et al. (Thoracic Surgery Dep, Central South Univ, Changsha, Hunan 410011, China, Email: wangxiang@csu.edu.cn) : Mirna-based biomarkers, therapies, and resistance in cancer. Int J Biol Sci 2020, 16(14), 2628-47.

MicroRNAs (miRNAs), small non-coding RNAs (ncRNAs) of about 22 nucleotides in size, play important roles in gene regulation, and their dysregulation is implicated in human diseases including cancer. A variety of miRNAs could take roles in the cancer progression, participate in the process of tumor immune, and function with miRNA sponges. During the last two decades, the connection between miRNAs and various cancers has been widely researched. Based on evidence about miRNA, numerous potential cancer biomarkers for the diagnosis and prognosis have been put forward, providing a new perspective on cancer screening. Besides, there are several miRNA-based therapies among different cancers being conducted, advanced treatments such as the combination of synergistic strategies and the use of complementary miRNAs provide significant clinical benefits to cancer patients potentially. Furthermore, it is demonstrated that many miRNAs are engaged in the resistance of cancer therapies with their complex underlying regulatory mechanisms, whose comprehensive cognition can help clinicians and improve patient prognosis. With the belief that studies about miRNAs in human cancer would have great clinical implications, we attempt to summarize the current situation and potential development prospects in this review.

3 illus, 2 tables, 227 ref

041503   PATEL V B, ACHARYA N (Pharmacognosy Dep, Nirma Univ, Ahmedabad - 382 481 Gujarat, Email: niyati20103@gmail.com) : Anti-urolithiatic activities of Macrotyloma uniflorum mediated through multiple pathway. Indian J Pharm Edu Res 2020, 54(2), 403-15.

Macrotyloma uniflorum Linn. (Fabaceae) seeds are widely used for their diuretic and urolithiatic effects in India. The present study investigated the effect of n-butanol fraction of M. uniflorum (nBFMU) on kidney stone using in vitro and in vivo method. Nucleation, crystal growth, crystal aggregation and crystal dissolution assays were performed for nBFMU. Two doses of nBFMU (400 and 800 mg/kg) were studied for their diuretic activity and sodium oxalate (NaOx) (70 mg/kg, i.p) induced urolithasis in male Wistar albino rats. Determination of body and kidney weight, measurement of various biochemical parameters in biological samples and examination of histology of kidney at the end of experiment were also done. nBFMU exhibited a concentration dependent inhibitory activity on nucleation and aggregation along with decreased number of crystals of calcium oxalate (CaOX) produced in metastable solutions of CaOX in the in vitro experiments. Co-administration of nBFMU with NaOX has significantly (p<0.001) increased the urine volume and the level of calculus inhibitors like magnesium, citrate and decreased the level of calculus promoters like oxalate, calcium, urea and uric acid. nBFMU supplement also prevented the pathological changes in kidney and increased the glomerulus activity of the kidney. These results indicate that nBFMU showed significant activity in urolithiasis which might be due to its diuretic, CaOX crystal formation inhibitory effects and its ability to increase the levels of inhibitors and decrease the level of promoters of urolithiasis.

5 illus, 8 tables, 45 ref

043295   GAN T, LI Y, ZHOU X-J, ZHANG H (Peking Univ First Hospital, Xicheng District, Beijing 100034, P.R China, Email: zhouxujie@bjmu.edu.cn) : Immunoproteasome in iga nephropathy: State-of-art and future perspectives. Int J Biol Sci 2020, 16(14), 2518-26.

IgA nephropathy (IgAN) is a leading cause of chronic kidney disease and renal failure. The exact pathogenesis of IgAN is not well defined, but some genetic studies have led to a novel discovery that the immunoproteasome probably plays an important role in IgAN. The immunoproteasome is a proteasome variant that is expressed when cells are stressed or receive inflammatory signals. While immunoproteasome is suggested to be mainly involved in major histocompatibility complex-I (MHC-I) antigen presentation, recent studies indicate that it may assert broad functions in trafficking events that activate both innate and adaptive immunity. In this review, we first summarize new insights into its functions in immunity, and discuss how it underlies its associations with IgAN. We also highlight its potential as a therapeutic target for the future.

3 illus, 1 table, 67 ref

043294   XU Q, FANG M, ZHU J, DONG H, CAO J, YAN L, LEONARD F, OPPEL F, SUDHOFF H, KAUFMAN A M, ALBERS A E, QIAN X (Clinical Laboratory Dep, Cancer Hospital of the Univ of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou, P.R. China, Email: qian_michelle2014@163.com (X.Q)) : Insights into nanomedicine for immunotherapeutics in squamous cell carcinoma of the head and neck. Int J Biol Sci 2020, 16(14), 2506-17.

Immunotherapies such as immune checkpoint blockade benefit only a portion of patients with head and neck squamous cell carcinoma. The multidisciplinary field of nanomedicine is emerging as a promising strategy to achieve maximal anti-tumor effect in cancer immunotherapy and to turn non-responders into responders. Various methods have been developed to deliver therapeutic agents that can overcome bio-barriers, improve therapeutic delivery into the tumor and lymphoid tissues and reduce adverse effects in normal tissues. Additional modification strategies also have been employed to improve targeting and boost cytotoxic T cell-based immune responses. Here, we review the state-of-the-art use of nanotechnologies in the laboratory, in advanced preclinical phases as well as those running through clinical trials assessing their advantages and challenges.

2 illus, 1 table, 17 ref

041500   HONMANE S M, CHIMANE S M, BANDGAR S A, PATIL S S (Pharmaceutics Dep, Shivaji Univ, Kolhapur - 416 112, Maharashtra, Email: sandiphonmane@gmail.com) : Development and optimization of capecitabine loaded nanoliposomal system for cancer delivery. Indian J Pharm Edu Res 2020, 54(2), 376-84.

The Main objective of this study was to develop and optimize Capecitabine loaded nanoliposomes for prolonged drug delivery in cancer treatment. Liposomes were prepared by the thin film hydration method followed by sonication. The parameters affecting the vesicle size and percentage drug entrapment of liposome are amount of soyaphosphatidyl choline and cholesterol used in their preparation. The Capecitabine liposomal formulation was optimized using 3² factorial design in this amount of soya Phosphatidylcholine and cholesterol were selected as two independent variables to obtain stable liposome with small vesicle size and maximum entrapment efficiency. Compatibility studies were carried out by using FT-IR and DSC, the results showed that there was no significant interaction between drug and excipients. The formulated liposomal prepartions were evaluated for various parameters and results were obtained for optimized batch (B3) Showed vesicle size 178.9nm, zeta potential -77.9mV to -82.7mV, entrapment efficiency 79.65 % and percentage drug release 92.07 % up to 12 h. Liposomal drug delivery is targeted as to provide more drug concentration at the site of action and with a sustainable drug release followed Higuchi-matrix model. Ultimately, reducing the dosing frequency with minimizing the side effects related to high drug intake. Liposome has been provided a spectrum of options and opportunities for designing and practicing site specific, targeted drug therapy.

9 illus, 4 tables, 29 ref

041499   PANDA S (MIT Coll of Pharmacy, Moradabad, Uttar Pradesh - 244 001, Email: drsubhran@gmail.com) : Formulation and evaluation by appling 3² (three squire) factorial design of lercanidipine hydrochloride buccal tablets with mucoadhesive polymers. Indian J Pharm Edu Res 2020, 54(2), 367-75.

The aim of the present research work is to develop buccal tablets of Lercanidipine hydrochloride to reduce dosage frequency; obtain optimized and controlled therapy, better patient compliance. Lercanidipine HCl was obtained as a gift sample from Sun Pharma, Baroda, India. Other ingredients like Na-alginate, Carbopol 934 P, Micro Crystalline Cellulose, Mannitol, Magnesium stearate, Ethyl cellulose and Hydroxy Propyl Methyl CelluloseK4M were purchased from various sources. All other ingredients used were of analytical grade. Attempt was made by using mucoadhesive polymers HPMC K4 M, sodium alginate and carbopol 934 P in combination with Ethyl Cellulose as an impermeable backing layer. Combination of polymer HPMC K4 M and sodium alginate release of drug was found in desired manner than other combinations. On the basis of the preliminary trials a 3² full factorial design was employed to study the effect of independent variables such as concentration of sodium alginate: HPMC K4M (X1) and type of filler (X2) on dependent variables. Factorial batches of F1 to F9 were formulated. HPMC K4M exhibited a much greater sustained effect on the release rate compared with sodium alginate. F4 shown the highest f2 value 70.46 and also all the h drug release was within the specified range. Based on the f2 value and targeted release profile the F4 batch was considered as optimized batch. Formulation F4 was subjected to an in vitro buccal permeation study. The results showed drug permeation of 99.04 % in 12 h. The correlation between in vitro drug release rate and in vitro drug permeation across the chicken mucosa was found to be positive, with a correlation coefficient (R²) of 0.9921. From kinetic modelling of the dissolution profile of the optimized formulation, it was concluded that there is erosion-controlled release of Lercanidipine from the buccal adhesive drug delivery system.

2 illus, 11 tables, 15 ref

041495   ACHARYA S D, TAMANE P K, KHANTE S N, POKHARKAR V B (Pharmaceutics Dep, Poona Coll of Pharmacy, Pune - 411 038, Maharashtra, Email: varsha.pokharkar@bharatividyapeeth.edu) : QbD based optimization of curcumin nanoemulsion: DoE and cytotoxicity studies. Indian J Pharm Edu Res 2020, 54(2), 329-36.

The aim of this study was to employ a Quality by design (QbD) based approach for the development and optimization of stable curcumin (CUR) nanoemulsion (NE). The NE was developed using Tween 80 as surfactant, Tocopheryl polyethylene glycol 1000 succinate (TPGS) as cosurfactant and Kollisolv MCT 70 as oil phase. The Design of Experiment (DoE) consisted of a 3 level- 2 factor full factorial design for the optimization. The independent variables were concentration of surfactant mix (Smix) and oil phase while particle size, zeta potential and polydispersity index (PDI) were the dependent variables. The NE was evaluated for particle size, zeta potential, pH, viscosity, transmittance, drug content, in-vitro drug release, stability and in vitro anticancer activity. The optimized formulation showed approximate particle size of 17nm, zeta potential -12.8 and polydispersity index 0.161 and was stable for three months at room temperature. The nanoemulsion showed better activity as compared to pure drug against pituitary and colon cancer cell-lines. QbD based approach can aid in the development of robust and thermodynamically stable NEs.

4 illus, 1 table, 26 ref

041494   SHETE D D, PATMAS M A, PATIL S P (Pharmacognosy Dep, SCES’s Indira Coll of Pharmacy, Pune - 411 033, Maharashtra, Email: minakshipatmas@gmail.com) : Pro-argin technology based formulation of eugenol containing toothpaste. Indian J Pharm Edu Res 2020, 54(2), 323-8.

Formation of cavities and hyper sensitivity are few dental problems, manifested by dental pain. Since long back, volatile oil present in clove buds (Myrtaceae) have been used as dental analgesic. Various toothpastes containing clove oil are marketed. Eugenol is chemically, terpenoids, active principle present in clove oil. It has several pharmacological potentials. Pro-argin technology is novel approach for formulation of toothpaste with arginine (8 %) and calcium carbonate. This research was aimed towards formulation of toothpaste containing eugenol by Pro-argin technology and its evaluation. Toothpaste containing eugenol was formulated with arginine (8 %) and calcium carbonate by simple trituration. Formulation was then evaluated for various parameters like pH, spreadability, foaming ability, content determination and also analysed for texture. On formulation, brownish, aromatic fragrant toothpaste was obtained with acceptable pH, spreadability and foaming ability. About 92.48 % of eugenol content of added quantity was determined. Eugenol can successfully be formulated in toothpaste by Pro-Argin approach. More than 90 % content determination would facilitate maximum release of added eugenol.

3 illus, 1 table, 21 ref

041493   KHANNA K, SHARMA D, KARWASRA R, SHARMA N, NISHAD D K, POPLI H, BHATNAGAR A (Pharmacy Dep, Research Univ, Vihar, New Delh, Email: popli.harvinder@gmail.com) : Intranasal nalbuphine formulation for faster management of pain in prehospital scenario; its safety and comparative efficacy in animal models. Indian J Pharm Edu Res 2020, 54(2), 310-22.

Nalbuphine (NLB) is an approved opioid analgesic for the management of severe pain in wounds, battlefield injuries and is recommended to subsidize labour pain during childbirth. The study aims to develop an intranasal opioid formulation for faster pain relief. Secondly to avoid patient inconvenience and make it available for buddy care. In case of war, accident or any natural calamities the feasibility of giving drugs from the parenteral route is not feasible as it requires high skilled medic/paramedic staff. Different formulations were made using NLB as an Active Pharmaceutical Ingredient (API), prepared formulations (F1-F10) were characterized and evaluated for various parameters. The formulation F5 was optimized as it met the desired criteria. F5 was further studied for its efficacy in an established pain model using Eddy’s hot plate method and pain scoring in animals. Pharmacodynamics study was conducted in Sprague dawley rats which were further verified by gamma scintigraphy using ^99mTc-pertechnetate labelled NLB showing significant and rapid deposition in the brain tissue. NLB nasal formulation was optimized successfully; at pH 6.4±0.10 and viscosity 2.5±0.13 (cps). Osmolarity and percent drug release of optimized nasal drop at 240 min was found to be 288±18 milliosmol/litre and 96.98±3.1 % respectively. Gamma scintigraphy study results revealed that the formulation (F5) was able to deliver NLB to the brain within 10 mins of administration and remained localized up to 240 min. Pre-clinical subacute toxicity profiling of optimized formulation was done in Sprague dawley rats at 3X dose. Data obtained from the study indicate that the developed NLB nasal formulation has better efficacy for pain management than conventional I.M injection.

9 illus, 7 tables, 42 ref

041491   DANDAGI P M, PANDEY P, GADAD A P, MASTIHOLIMATH V S (Pharmaceutics Dep, KLE Coll of Pharmacy (A constituent unit of KAHER), Belagavi - 590 010, Karnataka, Email: cool.pratibha25@gmail.com) : Formulation and evaluation of microemulsion based luliconazole gel for topical delivery. Indian J Pharm Edu Res 2020, 54(2), 293-301.

Drug delivery through topical route is mostly preferred for local dermatological action. Topical action of preparations has certain limitations in terms of drug solubility, residence time, lipophilicity and permeability. Conventional dosage forms such as creams, ointments etc exhibit drawbacks like problem in stability, stickiness, poor absorption as well as permeation mainly in case of large molecule. To overcome this, the origination of emulgel came into existence which basically focus on the delivery of hydrophobic drugs.¹ The present research is proposed to prepare and evaluate microemulsion based luliconazole gel for its enhanced solubility and permeability for better antifungal activity. The present study was aimed to prepare and evaluate microemulgel of luliconazole for topical application by using poly unsaturated fatty acids such as linseed oil. Sodium alginate was used as a gelling agent. Luliconazole microemulsion was prepared by varying the concentration of surfactants while the oil concentration was kept fixed. It was characterized for globule size and physical stability. Microemulgel was prepared and characterized for drug content, viscosity, spreadability, in vitro permeation study, skin irritation test, in vitro antifungal study and stability study. Skin irritation test was carried out after taking approval of Institutional Animal Ethics Committee. Two microemulsion were selected based on the globule area, PDI and physical stability and incorporated into the gel base of sodium alginate. The optimized microemulgel (F1) showed acceptable physical properties with lowest viscosity value (2325 cps) and showed pseudoplastic behaviour, highest spreadability value (3.7 gm.cm/sec), Highest drug permeation (63.50 %) after 5 hr. Skin irritation test showed no signs of irritation in any rats. Optimized formulation F1 represents greater zone of inhibition (38 mm) as compared to pure gel (34 mm) and showed acceptable stability profile. It can be concluded that luliconazole microemulgel prepared with linseed oil and sodium alginate can be a choice for topical antifungal treatment.

8 illus, 8 tables, 15 ref

046592   THOMAS A B, SHAIKH A S, RAJE A, LOKHANDE K B, SWAMY K V, NAGORE D H, DOKE R (Pharmaceutical Quality Assurance Dep, Dr. D.Y. Patil Institute of Pharmaceutical Sciences and Research, Pune, Maharashtra, Email: asha.thomas@dypvp.edu.in) : Herb drug interaction of Tinospora cordifolia (Willd.) Miers extract and Glimepiride: In vivo and in silico studies. Med Plants 2020, 12(4), 555-67.

Tinospora cordifolia (Willd.) Miers (TC), family Menispermaceae is a well-known traditional herb reported in traditional Indian literature for the treatment of various disorders such as diabetes and cardiovascular diseases. Berberine, the chief active constituent of TC is reported to possess anti-diabetic potential along with other isoquinoline alkaloids. TC and its active constituent berberine both have been reported to be potent CYP2C9 inhibitor, thereby raising herb drug interaction (HDI) potential on co-administration with drugs metabolized through the CYP pathway. The present study evaluated the pharmacokinetic HDI of TC hydroalcoholic extract with Glimepiride (GP) after oral co-administration in Wistar rats using in vivo pharmacokinetic and in silico studies. After oral co-administration of TC extract (100 mg/kg) with GP (20 mg/kg) in rats, drug concentrations in plasma were determined using HPLC method. The main pharmacokinetic parameters of C_max, T_max, AUC, Vd, CL, and MRT_0-t were calculated using Win Nonlin software. Also in silico molecular docking study using CYP2C9 (PDB ID: 1R90) as target protein and phytoconstituents (isoquinoline alkaloids) of TC as ligands were carried out using AutoDock Vina 1.1.2 to determine their CYP2C9 interaction potential. The PK interactions studies demonstrated an increase in the systemic levels of GP. A significant increase in C_max, AUC_0-t and MRT_0-t of GP (p<0.01) was observed, along with a substantial decrease in Vd and CL. In silico molecular docking studies on CYP2C9 demonstrated high inhibition potential of berberine (binding affinity: -9.6 kcal/mol) with formation of two hydrogen bonds with Ser 209 and Asn 474 in the active site of enzyme, complementing the literature reports as well as our in vivo PK findings. The results obtained from in vivo and in silico studies proposed that, for a co-administration of GP and TC extract, there exists a potentially significant PK HDI. This knowledge concerning potential HDI of GP might prove helpful for healthcare professionals as well as diabetic patients on GP therapy. It eventually warrants further studies to predict the pharmacokinetics and pharmacodynamics of GP in humans in case of HDI.

7 illus, 5 tables, 29 ref

046591   PANDEY M, BAJPAI M (GLA Univ, Mathura- 281 406, Email: minakshi.pandey_phd18@gla.ac.in) : Natural remedies for the treatment of arthritis: A review. Med Plants 2020, 12(4), 545-54.

Arthritis is a chronic joint disorder which imparts a major cause of disability globally. It usually targets the older population and arised due to improper regulation of pro-inflammatory cytokines and enzymes which lead to elevated levels of chemical mediators like prostaglandins, leukotrienes, nitric oxide, adhesion molecules and matrix metalloproteinase enzyme. Arthritis is treated by certain synthetic drugs which only provide symptomatic relief to the patients. Synthetic drugs have no any evidence to cure the disease, however they exhibits various toxic effects, side effects or recurring of symptoms on discontinuation of drugs. Therefore, it is an urgent need of alternative treatments against arthritis. On view of these facts, this review emphasizes the medicinal plants, herbal supplements and spices, herbal formulations and some traditional approach to treat arthritis.

1 illus, 2 tables, 56 ref

041490   SHARMA A, SHARMA S, TOMAR D S (R.J Coll of Pharmacy, Aligarh - 202 001, Uttar Pradesh, Email: xs2akhil@gmail.com) : Optimization of in-situ nanoparticulate gel of ofloxacin using factorial design to improve treatment strategy for conjunctivitis and corneal ulcers. Indian J Pharm Edu Res 2020, 54(2), 284-92.

One of the main problems in ophthalmic drug delivery is the rapid elimination of conventional liquid eye drops from the eye due to rapid tear turnover resulting precorneal loss because of irritation caused by the drug preparation from large volume of the administered eye drop (~50 µl), a very small amount i.e. <5 % of the dosage actually penetrates and is able to reach intraocular tissues. Ofloxacin (OFL), a fluoroquinolone drug is used for the treatment of conjunctivitis and corneal ulcers and it is given quarterly in a day because its bioavailability is very low. The present study was designed to increase the ocular residence time of drug by formulating nanoparticulate in-situ gel for ensuring low irritation, better release and compatibility with ocular tissue. Initially, OFL nanoparticles were prepared using single emulsion solvent evaporation method and evaluated for various evaluation parameters. Based on the results, an optimized batch (B3) was selected by applying factorial design; which was further converted into in-situ gel. Results of evaluation parameters revealed that optimized batch (B3) showed significant (p=0.0001 and p=0.0090) increased in particle size and entrapment efficiency in comparison to rest batches. Moreover, nanoparticulate in-situ gel showed satisfactory results. Hence, Ofloxacin nanoparticulate in-situ gel thus formulated showed controlled drug release with lesser dosing frequency.

12 illus, 9 tables, 17 ref

041489   WEN J, LU W, CHEN Z (Pharmacology Dep, Anhui Medical Univ, Anhui Province, China, Email: chpharmzw@163.com) : Innovation and construction of examination database of pharmacology. Indian J Pharm Edu Res 2020, 54(2), 279-83.

Pharmacology is one of the cornerstones in medical curricula, besides, examination is an effective way to evaluate the teaching quality and student outcome. This study was undertaken to demonstrate the examination innovation and construction of online exam questions bank of Pharmacology. The pharmacological exercises compiled previously were attached with standarded parameters and used as main source of online exam questions bank, the multifaceted parameters of questions were determined for easy to construct test paper. We next sought to apply this new established online exam database to the fourth semester students of nursing and the fifth semester students of clinical medicine. We successfully established an online exam questions bank of 3000 questions containing single choice, multiple choice and true/false and covering all of knowledge points in syllabus. And then, we successfully formed the exam papers from online questions bank abide by the examination syllabus and applied to the fourth semester students of nursing and fifth semester students of clinical medicine. The exam scores were respectively higher than those of sixth semester students of nursing and the seventh semester students of clinical medicine who used traditional examination method last year. These findings indicated that the new established online exam questions bank may better promote the teaching quality and student outcome and lay the foundation for the further implementation of the online exam.

2 illus, 1 table, 13 ref

043293   LÓPEZ-COLLAZO E, AVENDAÑO-ORTIZ J, MARTÍN-QUIRÓS A, AGUIRRE L A (La Paz Univ Hospital, Madrid 28046, Spain, Email: elopezc@salud.madrid.org) : Immune response and COVID-19: A mirror image of sepsis. Int J Biol Sci 2020, 16(14), 2479-89.

The emergence of SARS-CoV-2 virus and its associated disease COVID-19 have triggered significant threats to public health, in addition to political and social changes. An important number of studies have reported the onset of symptoms compatible with pneumonia accompanied by coagulopathy and lymphocytopenia during COVID-19. Increased cytokine levels, the emergence of acute phase reactants, platelet activation and immune checkpoint expression are some of the biomarkers postulated in this context. As previously observed in prolonged sepsis, T-cell exhaustion due to SARS-CoV-2 and even their reduction in numbers due to apoptosis hinder the response to the infection. In this review, we synthesized the immune changes observed during COVID-19, the role of immune molecules as severity markers for patient stratification and their associated therapeutic options.

2 illus, 2 tables, 17 ref

043292   CHEN H, HU X, YANG R, WU G, TAN Q, GOLTZMAN D, MIAO D (Nanjing Medical Univ, Nanjing, Jiangsu, 210029, The People’s Republic of China, Email: dsmiao@njmu.edu.cn) : SIRT1/FOXO3a axis plays an important role in the prevention of mandibular bone loss induced by 1,25(OH)_{2D deficiency.} Int J Biol Sci 2020, 16(14), 2712-26.

It has been reported that 1,25 dihydroxyvitamin D [1,25(OH)₂D] deficiency leads to the loss of mandibular bone, however the mechanism is unclear. We investigated whether the Sirt1/FOXO3a signaling pathway is involved in this process. Using a 1,25(OH)₂D deficiency model induced by genetic deletion in mice of 25-hydroxyvitamin D-1α hydroxylase [1α(OH)ase^-/- mice]. We first documented a sharp reduction of expression levels of Sirt1 in the 1α(OH)ase^-/- mice in vivo. Next, we demonstrated dose-dependent upregulation of Sirt1 by treatment with exogenous 1,25(OH)₂D₃ in vitro. We then identified a functional VDR binding site in the Sirt1 promoter. By crossing Prx1-Sirt1 transgenic mice with 1α(OH)ase^-/- mice we demonstrated that the overexpression of Sirt1 in mesenchymal stem cells (MSCs) greatly improved the 1α(OH)ase^-/- mandibular bone loss phenotype by increasing osteoblastic bone formation and reducing osteoclastic bone resorption. In mechanistic studies, we showed, in 1α(OH)ase^-/- mice, decreases of Sirt1 and FoxO3a, an increase in oxidative stress as reflected by a reduction of the antioxidant enzymes peroxiredoxin1 (Prdx1), SOD1 and SOD2 expression, and an increase of markers for osteocyte senescence and senescence associated secretory phenotypes (SASP), including β-galactosidase (β-gal), p16, p53 and p21. The targeted overexpression of Sirt1 in the 1α(OH)ase^-/- mice restored the expression levels of these molecules. Finally, we demonstrated that a Sirt1 agonist can upregulate FOXO3a activity by increasing deacetylation and nuclear translocation. Overall, results from this study support the concept that targeted increases in Sirt1/FOXO3a signaling levels can greatly improve the bone loss caused by 1,25(OH)₂D deficiency.

6 illus, 1 table, 51 ref

041487   HAZRA A, BOSE P, PATTANAYAK S P (Pharmaceutical Sciences and Technology Dep, Birla Institute of Technology, Ranchi - 835 215, Jharkhand, Email: sppattanayak@bitmesra.ac.in) : Epidermodysplasia verruciformis: Functional role of e6 and e7 oncoproteins as the link between viral infection to carcinogenesis. Indian J Pharm Edu Res 2020, 54(2), 251-63.

Cancer is the most dreadful disease since last few decades. Besides all causative reasons, it is observed that one in every five cancers globally is linked with infectious diseases. Among all the infectious oncogenic viruses, 35 % of the human cancers have been caused by Human Papilloma Viruses (HPVs). One such type is Epidermodysplasia Verruciformis (EV), a tropical disease also known as the Tree man syndrome. EV is one of the most potential illustrations linking viral infection with skin carcinogenesis. The HPV virus acts as a cofactor with the UV radiation. The lesions caused by invasion of the HPV in the sun exposed wounded areas of the EV patients clearly links βHPV and cellular proliferation where EVER1 and EVER2 gene mutations are equally cardinal. When the viral infection persists, the somatic mutations over time accumulate, leading to precancerous lesions at first, followed by the malignant transformation. Two out of the eight proteins that the HPV genome codes for namely E6 and E7, account for the high-risk type HPVs that cause cancer. Both work similarly but on different sites. Both E6 and E7 proteins encode the main HPV's oncoprotein which promotes cell progression and viral replication. The role of E6 and/or E7 in tumor progression, metastasis promotion, NOTCH and TGFβ pathway modification, DNA methyltransferase stimulation, histone modification, E-cathedrine modification etc. have been described in this review for the better understanding of the linkage between viral infection and carcinogenesis process.

9 illus, 52 ref