Microbial lipase,one of important industrial biocatalysts,has been used widely in many industrial and agricultural fields.It is always the research focus to screen,mine and develop the microbial lipases with novel catalytic activity and high stability.This paper introduces briefly the pathways and methods to mine novel microbial lipase resources from six aspects,including extremophile,metagenome,genome database,protein engineering,immobilization,chemical modification,etc.

Objective:Preparation and immune characteristic analysis of polyclonal antibody against hypervariable region protein of Taura syndrome virus major capsid protein VP 1 as a reference for studies on immunological diagnosis reagent.Methods:The recombinant vector pET-VP1 was transformed into E.coli BL21 for protein expression.Immunizing a New Zealand rabbit with purified VP1 protein,the titer of anti-VP1 serum was determined by Agar diffusion test and ELISA.Monoclonal phage specific binding to the purified VP1 protein was used for competitive inhibition test.Results: The VP1 protein was soluble and high expression in E.coli BL21.The biological activity titer of anti-VP1 serum reached 1∶26 ,1∶217 determined by Agar diffusion test and ELISA respectively.A litter binding activity of antiserum and VP 1 protein could be blocked by monoclonal phage , but would not affect the final positive result.Conclusion:High titer antibody Preparation of the VP 1 hypervariable region protein.The binding activity of the polyclonal antibody with VP1 protein was not affected by the mutations of VP 1 protein in minority areas ,so the antiserum could be used as immu-nological detection diagnosis agent.

Objective To investigate the expression and clinical features of amplified in breast cancer 1 (AIB1) and epithelial mesenchymal transition (EMT) markers in human hepatocellular carcinoma and to observe the effect of AIB1 silencing by RNA interference (RNAi) on expression of EMT markers and invasiveness of HepG2 cells.Methods In this study,expression of AIB1,E-cadherin,Vimentin,ZO-1,and N-cadherin protein in 81 hepatocellular carcinomas were assessed through immunohistochemistry and clinicopathological significance was analyzed.After the lentiviral vector of AIB1 RNA interference was transfected into HepG2 cells,the expression of AIB1 and EMT markers was detected by real-time PCR and Western blot.The invasion and metastasis was evaluated by Transwell analysis.Results The expression of AIB1 protein was significantly up-regulated in the hepatocellular carcinoma tissue compared to the normal tumor adjacent tissue.The frequency of AIB1 overexpression in hepatocellular carcinomas with lymph node metastasis is 63％ (P ＜ 0.05).Correlation analysis demonstrated that the AIB1 protein expression was inversely correlated with E-cadherin,and positively correlated with Vimentin in hepatocellular carcinomas.After transfection with AIB1 targeting siRNA,the expression of AIB1 mRNA and protein decreased significantly (P ＜ 0.05).Knockdown of AIB1 expression increased the expression of E-cadherin and inhibited the expression of Vimentin.In addition,the invasion of HepG2 cells silenced AIB1 were significantly descented.Conclusion Above data suggests that overexpression of AIB1 might promote invasiveness and metastasis of cancer cells through regulation of E-cadherin and Vimentin expression in hepatocellular carcinomas.

We used metabolomics to investigate the ability of a traditional Mongolian medicine called modified Tabusen-2 (MT-2) to improve kidney yang deficiency (KYD) in rats. All animal experiments were conducted under the guidance and standards of the Medical Ethics Committee of Inner Mongolia Medical University. SD rats were divided into 6 groups of six rats: a normal group, a model group, Jinkuishenqi pill administration group (1.26 g·kg^-1), and MT-2 administration in high-, medium- and low-dose groups (1.512, 0.756, and 0.378 g·kg^-1). KYD was established by intramuscular injection of hydrocortisone (HC) and biochemical indicators and clinical characterization was used to confirm that KYD was established. All groups received intragastrically administered drug (Jinkuishenqi pill or MT-2) or saline. Serum from each group was collected after 8 weeks and analyzed by UPLC-Q-exactive-MS to measure various biochemical indicators. The biomarkers affected by MT-2 were identified and the metabolic pathways of KYD regulated by MT-2 were analyzed by metabolomic analysis. The results show that MT-2 can decrease serum creatinine (Cr) in KYD rats and significantly increase (P<0.05) the content of thyroid stimulating hormone (TSH) and luteinizing hormone (LH). In serum samples, 38 biomarkers such as corticosterone, L-phenylalanine, and DL-tryptophan were measured as possible indicators for disease development in KYD rats. MT-2 lowered 18 biomarkers of KYD, including corticosterone, deoxycorticosterone, and L-phenylalanine, and altered 13 related metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and steroid hormone biosynthesis, resulting in an overall improvement in KYD. MT-2 appears to be important in improving KYD in rats mainly by regulating metabolites such as amino acids, steroids and lipids.

Objective To compare the application of the 2013 Chinese Diabetes Society (CDS 2013) criteria, the 2009 joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention(JIS)criteria, the 2005 International Diabetes Federation(IDF)criteria and the 2005 US National Cholesterol Education Program Adult Treatment PanelⅢ (ATPⅢ) criteria among the elderly in Nanjing. Methods A total of 1 832 elderly (≥60 years old) were recruited in Nanjing using cluster random sampling and were surveyed with questionnaires, physical examination, and laboratory test between March and December 2013. The different prevalence under CDS 2013, JIS, IDF and ATPⅢ criteria were calculated respectively. SPSS 19.0 was used to conduct t test, chi-square test and kappa test to compare the prevalence of metabolic syndrome and the consistency of four diagnostic criteria in the elderly. Results According to CDS 2013, JIS, IDF, and ATPIII, prevalence of MS in the elderly in Nanjing was 39.9%, 58.2%, 46.5%, and 54.5%, respectively. The highest value of kappa was 0.920 between JIS and ATPⅢ, with a compliance rate of 96.1%．The value of kappa was 0.841 between ATPⅢ and IDF, with a compliance rate of 92.0%. Conclusions The prevalence of metabolic syndrome among the elderly in Nanjing is relatively high under different criteria. The higher detection rate of MS by the JIS standard and ATPIII standard can help to find MS earlier and reduce the cardiovascular damage.

OBJECTIVETo investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.

METHODThe effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.

RESULTAfter 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.

CONCLUSIONCur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.

Cell Cycle ; drug effects ; radiation effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Curcumin ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; radiation effects ; Humans ; RNA, Long Noncoding ; genetics ; Radiation Tolerance ; drug effects

OBJECTIVETo screen and identify proteins that interact with hepatitis C virus NS3 by means of T7-phage display system.

METHODSHepatitis C virus NS3 was expressed by prokaryotic expression and used as a selected molecule to biopan the T7 select human liver cDNA library; the selected positive clones were identified using DNA sequencing and analyzed with BLAST program in GenBank.

RESULTSAfter BLAST analysis in all the positive clones, the proteins which interacted with the hepatitis C virus NS3 were found to be serpin peptidase inhibitor, clade A, member 1 (SERPINA1) and cyclophilin-LC.

CONCLUSIONT7-phage display system is a convenient, rapid and effective method for screening interacting proteins. The proteins thus selected will provide an important means for studying the pathogenesis and carcinogenesis of HCV.

Cell Line ; Gene Library ; Hepacivirus ; metabolism ; Humans ; Peptide Library ; Protein Interaction Mapping ; methods ; Viral Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

Objective To understand the risk situation of imported schistosomiasis and its main influencing factors in Shang-hai City from 2005 to 2015. Methods A retrospective survey was performed,and the database was established based on the da-ta collection of imported schistosomiasis,Oncomelania hupensis snail situation and mobile population in Shanghai City from 2005 to 2015. Results From 2005 to 2015,the mobile population in Shanghai City increased by 123.92％,which reached about 9816500,and 54.70％of them were located in the inner suburban districts. The accumulated areas with snails of 7.13 hm2 were found in 16 towns of 4 outer suburbs(excluding Chongming District). A total of 23 cases of imported schistosomiasis were found in Shanghai City,and the number of the imported schistosomiasis cases was on the downward trend(rs=-0.782,P=0.004). From 2005 to 2009,11 out of 13 imported schistosomiasis cases(84.62％)were distributed in the inner suburban dis-tricts where no O. hupensis snails were found at the same time. From 2010 to 2015,9 out of 10 imported schistosomiasis cases (90％)were distributed in the outer suburban districts where O. hupensis snails were found at the same time. Conclusion The number of imported schistosomiasis cases in Shanghai is on the downward trend. However,the threat of imported schistosomiasis to the prevention and control of schistosomiasis in Shanghai City should not be ignored.

OBJECTIVETo assess the effect of different schemes of premenopausal tamoxifen therapy on the endometrium.

METHODSTotally 109 normal premenopausal women positive for high-risk factors of breast cancer were divided into two groups, namely periodic and consecutive tamoxifen treatment groups. Endometrial thickness as examined by vaginal sonography was assessed in relation to duration of tamoxifen use and time from discontinuation of the drug.

RESULTSAfter one year of tamoxifen use, the mean endometrial thickness in periodic treatment group was 6.5-/+1.4 mm, and 10.2-/+2.0 mm in consecutive treatment group. Endometrial thickness increased with the duration of tamoxifen use at the rate of 0.51 mm/year in the periodic treatment group, and 0.73 mm/year in consecutive treatment group. After discontinuation of tamoxifen, the endometrial thickness in the former group decreased by 1.29 mm/year, and by 1.33 mm/year in the latter.

CONCLUSIONSEndometrial hyperplasia is obviously milder in premenopausal women receiving periodic tamoxifen treatment who are at risk for breast cancer than that in women with consecutive treatment. After discontinuation of the drug, the endometrial thickness decreases at a roughly equal slow rate in the two groups.

Adult ; Breast Neoplasms ; drug therapy ; Drug Administration Schedule ; Endometrium ; anatomy & histology ; diagnostic imaging ; drug effects ; Female ; Humans ; Premenopause ; drug effects ; Risk ; Tamoxifen ; administration & dosage ; pharmacology ; therapeutic use ; Time Factors ; Ultrasonography

BACKGROUNDEsophageal cancer is the sixth most common cause of cancer-related death worldwide. Prior studies had demonstrated potential synergistic antitumor activity of gemcitabine in combination with cisplatin. Therefore, we studied the efficacy and tolerability of such combination for esophageal cancer.

METHODSBetween October 2003 and October 2006, thirty-eight patients with metastatic or recurrent advanced squamous cell carcinoma of the esophagus were enrolled. The median number of treatment cycles per patient was 4 (range 1 - 7). Gemcitabine was given at 1000 mg/m(2) over 30 minutes on days 1, 8 and cisplatin 40 mg/m2 was given on days 1, 2 in an every 21-day cycle.

RESULTSThe median follow-up for all 38 patients was 76 months (range 11 - 88 months). The overall response rate was 42.1% (95%CI, 25.5% - 56.5%). Median progression-free survival and median survival for all patients were 4.1 months (95%CI, 3.0 - 5.7 months) and 10 months (95%CI, 7 - 12 months), respectively. Patients with a response had significantly longer median survival compared with the patients without a response (11 months vs. 7.5 months, P = 0.0069). Overall survival at 1 year was 36.8%, at 2 years was 10.5%, and at 5 years was 5.3%. The most common grade 3 - 4 toxicity for all patients was leucopenia (44.7%).

CONCLUSIONSThis cisplatin-gemcitabine regimen was manageable and had significant efficacy in patients with esophageal squamous cell carcinoma. Patients with a response had improved survival time. Furthermore, a small number of the patients with metastatic esophageal cancer were still alive in 5 years with this regimen.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; Carcinoma, Squamous Cell ; drug therapy ; mortality ; Cisplatin ; therapeutic use ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Esophageal Neoplasms ; drug therapy ; mortality ; Female ; Humans ; Male ; Middle Aged