Antigens, CD20 ; metabolism ; CD79 Antigens ; metabolism ; Carcinoma, Neuroendocrine ; metabolism ; pathology ; Carcinoma, Renal Cell ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Middle Aged ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Nephrectomy ; PAX5 Transcription Factor ; metabolism ; Wilms Tumor ; metabolism ; pathology

OBJECTIVE:To establish UV-spectrophotometry method in the determination of metformin hydrochloride in compound metformin hydrochloride tablets.METHODS:The determination was carried out with water as the solvent under the detection wavelength of233nm.RESULTS:There was a good linear relationship between metformin hydrochloride and its absorbability(r=0.9999)within the concentration range of1～10?g/ml.The average recovery was99.3%with RSD at0.24%(n=9).CONCLUSION:The method is accurate,fast,convenient,and which can be used to determine the contents of metformin hydrochloride in compound metformin hydrochloride tablets directly without isolation.

To investigate the induction of apoptosis of mouse colonic adenocarcinoma CT26 cells by recombinant Clostridium difficile toxin B (rTcdB), CT26 cells were exposed to different concentrations of rTcd B. Inhibition of cell proliferation was measured by MTT assay. The activation of Caspase 3 was measured by colorimetric method. Cell morphological analysis and flow cytometry were performed to confirm cell apoptosis. rTcd B inhibited the proliferation of CT26 cells in a timeand dose-dependent manner. Caspase 3 activity in CT26 cells was elevated remarkably after rTcd B exposure for 6 h, 12 h, 18 h or 24 h, as compared with the control group. Morphological changes were observed by fluorescence microscopy. The exposure of rTcd B to CT26 cells induced a timeand dose-dependent apoptotic cell death as determined by flow cytometry analysis. The results showed that recombinant Clostridium difficile toxin B induced apoptosis of CT26 cells.

Objective To investigate the effect of folic acid on the DNA methylation of tumorrelated genes promoters in healthy human peripheral blood mononuclear cells(PBMC). Methods Ten healthy volunteers were divided into two groups, and were randomized to receive either 5 mg folic acid (n=5)or placebo(n = 5) , one time per day for 3 months. The serum folic acid concentration was detected with chemiluminescence enzyme immunoassay kit before and after the intervention. The methylation statuses of five tumor-related genes promoter, including oncogenes c-myc, c-Ha-ras,tumor suppressor genes p16INK4A, E-cadherin and mismatch repair gene hMLH1 in PBMC were detected by bisufite sequencing. Results After folic acid intervention, the level of serum folic acid increased significantly in intervention group (t= -4. 739,P<0. 05) , however no significant difference in control group. After three-month folic acid intervention, the level of methylation of oncogene c-myc promoter increased from 4%, 3. 3%, 4. 1% before intervention, one week after intervention, one month after intervention respectively to 8%(t= -4. 079,P<0. 05), while no significant change in placebo taken group. Before and after the folic acid intervention, there was no significant difference of DNA methylation of other tumor-related genes promoter, including c-Ha-ras、E-cadherin、p16INK4Aand hMLH1. Conclusion Folic acid intervention can up-regulate DNA methylation of oncogene c-myc promoter, but can not affect the promoter methylation status of tumor suppressor genes E-cadherin,p16INK4Aand hMLH1.

Objective To study the effects of extracellular-signal regulated kinase mitogenactivated protein kinase (ERK-MAPK) signaling pathway inhibition on histone phosphorylation and the related gene expression in human colorectal cancer cells.Methods Two human colorectal cancer cell lines (SW1116 and HCT116) were cultured and treated with gradient(0,20,40/μmol/L) doses of ERK-MAPK signaling pathway inhibitor U0126.Cell viability was determined by cell counting kit 8 (CCK-8) assay.Cell cycle distribution was assessed by flow cytometry.The expression levels of histone H3 kinases including ribosomal S6 serine-threonine kinase (RSK-2) and mitogen-and stressactivated protein kinase 1 and 2 (MSK1 and MSK2),and the levels of histone H3 (Ser10) phosphorylation and c-Fos protein were detected using Western blotting.Results Treatment of these two human colorectal cancer cell lines with ERK-MAPK inhibitor resulted in a time and dose-dependent inhibition of cell proliferation significantly. Proliferation rate of HCT116 was reduced to 47% at 72 hours after 40/μmol/L U0126 treatment. Cell cycle analysis showed that the percentage of phase G0/G1 cells significantly increased (P<0. 01) and the percentage of phase S cells decreased (P<0.01) after treatment with ERK-MAPK inhibitor. The expression of MSK1 and RSK2 reduced obviously in both of human colorectal cancer cell lines treated with U0126, which resulted in a 28% and 40% reduction of levels of MSK1 and RSK2 as compared with control HCT116 cells respectively,while no detectable change in the expression of MSK2 was found. Consistent with this, the expression level of histone H3 (ser10) phosphorylation was markedly down-regulated by ERK-MAPK inhibitor, and the related protein c-Fos expression decreased accordantly. Conclusions Decreased ERK-MAPK signaling pathway may reduce histone H3 (Ser10) phosphorylation via suppression of the activity of histone H3 kinase including MSK1 and RSK2, but not MSK2, consequently decrease the expression of c-Fos protein, which results in the inhibition of colorectal cancer cells proliferation.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Objective:To express secreted protein-pgp3 of Chlamydia trachomatis(Ct)plasmid,produce monoclonal antibodies (mAbs)and identify their basic biological characteristics.Methods: Construction pGEX-6p2-pgp3 prokaryotic expression vector,then expressed GST-pgp3 fusion protein in E.coli as antigen used to immune BALB/c mice, spleen cells were fused with SP2/0 mouse myeloma cells.The hybridoma cell lines of screening mAbs were secreted by ELISA,and mAbs specificity,type,class and titer were identified.Results:GST-pgp3 fusion protein was successful expressed,5 strains stable hybridoma cell lines that secreted mAbs were screened out,including 4 strains secreted anti-pgp3 mAbs(P1B3,P2A1,P2B6,P2C2),mAbs type were IgG1/κ,the other strain secretion anti-GST mAbs(P3B4),mAbs type was IgG2b/κ.The titer of P1B3,P2A1,P2B6,P2C2,P3B4 were 1∶6 400,1∶3 200,1∶12 800,1∶6 400 and 1∶6 400 respectively.Conclusion:Successful prepared anti-pgp3 mAbs,and lay a foundation for further study the function of Ct plasmid protein pgp3 and the establishment of Ct detection method.

Objective To discuss the effect of light invasive treatment of endometrial in husband sperm intrauterine insemination outcomes.Methods A total of 248 patients receiving husband sperm intrauterine insemination were divided into two groups by random digits table method.The patients in light invasive treatment of endometrial group (138 cases with 246 cycles) were performed with light invasive treatment of endometrial,and the patients in control group (110 cases with 201 cycles) were not perfomed with light invasive treatment of endometrial.Human chorionic gonadotropin day of endometrial thickness,intrauterine insemination tube placed successfully or not,whether fertilization tube bloody and clinical pregnancy rates and so on were compared between two groups.Results There was no significant difference in human chorionic gonadotropin day of endometrial thickness between two groups (P ＞ 0.05).The success rate of intrauterine insemination tube placement in light invasive treatment of endometrial group was 78.5％ (193/246),which was higher than that in control group (65.2％,131/201),and there was significant difference (P ＜ 0.05).The intrauterine insemination tube bloody rate in light invasive treatment of endometrial group was 13.4％ (33/246),which was lower than that in control group (34.8％,70/201),and there was significant difference (P ＜ 0.05).The pregnancy rate in light invasive treatment of endometrial group was 31.9％ (44/138),which was higher than that in control group (18.2％,20/110),and there was significant difference (P ＜ 0.05).There were no significant differences in ectopic pregnancy rate and abortion rate between two groups (P ＞ 0.05).Conclusion Through light invasive treatment of endometrial the husband sperm intrauterine insemination outcomes can be improved.

Purpose:To study the influence of treatment with high dose progesterone on telomerase activity in patients with endometrial adenocarcinoma.Methods:20 patients were treated for adenocarcinoma of endometrium with high dose progesterone and the change of telomerase activity in tumor tissue was examined by PCR-TRAP ELISA before and after treatment.Results:The average A value of tumor telomerase before and after treatment were 0.3636(0～1.11) and 0.1134 (0～0.556),respectively.It shows that telomerase activity in tumor was decreased by treatment with progesterone (P

Objective To investigate the application validity of the adjusted norm of WAIS-RC in the identification of intellectual disability.Methods128 patients with mental retardation (MR) were chosen for identification of the intellectual disability, measured their full scale IQs (FIQ) according to the original norm, calculated their four-subtest short forms FIQ respectively by Tellegen way according to the original standard norm and the new adjusted norm, then compared and analyzed the outcomes.ResultsThe short forms FIQ according to the original and the new norm had high correlation to full forms FIQ (P<0.01). The average of the short forms FIQ was higher than full forms FIQ according to the original norm (P<0.01), showing no significant difference according to the new norm(P＞0.05). In severe intelligence defected group according to full forms IQ, the grade classification corresponding rate of short forms FIQ according to the new norm was 0.00%, as well as the original norm. That in medium and mild intelligence defected groups was higher than that of original norm(P＜0.01).ConclusionThe test validity of adjusted norm short forms of WAIS-RC is superior to the original norm, but not suitable for severe intelligence defected.