Objective: To express the fusion protein of hOct4 and cell penetrating peptides using prokaryotic expression systems, and to optimize its expression methods and observe the membrane penetrating ability of the fusion proteins. Methods: The pET-based prokaryotic expression system was constructed by genetic engineering, and the fusion plasmid was transferred into E. coli BL21(DE3) and Rosetta2(DE3). The protein was purified by Ni affinity chromatography and identified by Western blotting analysis. The penetrating ability of the Rhodamine-labelled fusion protein was investigated using BJ cells. Results: We successfully constructed pET21a(+)-hOct4-11R-His and pET21a(+)-EGFP-11R-His vectors. Fusion proteins hOct4-11-His and EGFP-11R-His were generated by transfering the plasmids into E. coli. The fusion protein was verified by Western blotting analysis and was detected in BJ cells. Conclusion: We have successfully generated EGFP-11R-His and hOct4-11R-His fusion proteins, and the proteins can effectively enter the BJ cells and locate around the nuclei.

Hepatitis virus NAT reagents are now widely used clinically. However, the qulity of domestic and foreign NAT reagents varies dramatically. The main reasons for these differences including the manufacture technique, test principle and assay procedure were discussed in this paper and current status of the quality control of the NAT reagents were also described. Finally, it was pointed out that strengthening public supervision and laboratory internal control are very important for the quality improvement of the domestic reagents.

Apoptosis ; Down-Regulation ; Electron Transport Complex IV ; chemistry ; genetics ; metabolism ; Humans ; Mitochondria ; metabolism ; Mutation ; Neoplasms ; genetics ; metabolism ; pathology

Aneuploidy ; Gene Rearrangement ; Glutathione S-Transferase pi ; metabolism ; Humans ; Male ; Methylation ; Neoplasm Grading ; Neoplasm Staging ; Neoplasms, Multiple Primary ; genetics ; metabolism ; pathology ; surgery ; Oncogene Proteins, Fusion ; genetics ; Prostate-Specific Antigen ; metabolism ; Prostatic Intraepithelial Neoplasia ; genetics ; pathology ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery

Objective To investigate the role of cancer stem cells in radiation resistance of esophageal cancer and its molecular mechanism, and to provide a theoretical basis for radiotherapy for esophageal cancer.Methods Esophageal cancer cell line TE1 was treated with 8 Gy of radiation. Esophageal cancer cell line with resistance to radiation, TE1-res, was established and screened.Cell counting was used to evaluate cell proliferation.Flow cytometry was used to determine the expression of CD44 (high) CD24(-) CD133(+) and apoptosis in cells.The colony formation assay was used to determine the colony-forming rate and cell survival curve.Bisulfite sequencing PCR was used to determine the methylation status of cancer suppressor genes.Comparison of the data was made by group t test or analysis of variance. Results Compared with TE1 cells, TE1-res cells had significantly enhanced proliferation, a significantly higher proportion of CD44( high) CD24(-) CD133(+) cells, and significantly enhanced resistance to apoptosis (mean value 20.84×105 vs.4.46×105/day, P=0.008;(38.0±2.9)%vs.(10.1±1.3)%, P=0.001;mean value 33.23% vs.10.50%, P=0.003 ) .After treatment with 8 Gy of radiation, TE1-res cells had significantly higher colony-forming rate and D0 value than TE1 cells ((14.3±2.6)%vs.(0.9±0.3)%, P=0.011;3.28 vs.2.19 Gy, P=0.125 ) .Moreover, the promoter methylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B was significantly enhanced in TE1-res cells than in TE1 cells ((89.7±4.9)%vs.(5.0±0.5)%, P=0.001;(92.3±4.7)%vs.(10.4±0.7)%, P=0.001;(90.7±3.7)%vs.(7.9±0.4)%, P=0.001;(83.4±5.7)%vs.(17.2±1.2)%, P=0.002;(90.2±
6.7)%vs.(4.4±1.2)%, P=0.002).Conclusions Cancer stem cells play an important role in radiation resistance of esophageal cancer. The resistance to radiation is closely associated with promoter hypermethylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B.

Objective To establish a radioresistant esophageal squamous cancer cell line,and to identify the radioresistant genes and mechanisms.Methods The radioresistant KYSE410-res cell line was established by repeated exposure of cell line KYSE410 to radiation.The proliferation and apoptosis of esophageal squamous cancer cells were evaluated before and after radiation.The changes in gene expression of the esophageal squamous cancer cells after radiation were determined by gene microarray and analyzed by group t test.The genes with significant difference in expression after radiation were validated.Results The KYSE410-res cells had significantly enhanced proliferation and anti-apoptosis than the KYSE410 cells (all P＜0.05).The result of gene microarray showed that compared with the KYSE410 cells,the KYSE410-res cells had the expression of 463 and 251 genes upregulated and downregulated by no less than 4 folds,respectively.Those genes with different expression levels after radiation were mainly responsible for cell proliferation,adhesion,signal transduction,angiogenesis,reactive oxygen metabolism,cell damage repair,and the MAPK/ERK signaling pathway.OAS2 and UBD were key proteins in the network.In the KYSE410-res cells,the expression of HLA-DQBI,MMP1,NCAM1,ZNF521,GPC6,SELENBP1,LCN15,and TFPI-2 genes measured by real-time PCR was consistent with that measured by gene microarray.Conclusions Abnormal activation of the MAPK/ERK signaling pathway,upregulated expression of OAS2 and UBD,downregulated expression of TFPI-2,and upregulated expression of MMPs may play a role in radioresistance of esophageal cancer cells.

OBJECTIVE:To determine the content of ?-schizandrin,one of the effective ingredients,in Shuangjia Wuling capsules METHODS:The RP-HPLC method was performed with YWG C18 column(4 6mm?250mm) The mobile phase consisted of methanol-water(72∶28) The detecting wavelength was 254nm RESULTS:The calibration curve for ?-schizandrin was linear in the range of 0 0 207～0 4 130mg/ml(r=0 9 999) The average recovery was 97 68% with RSD=1 85% CONCLUSION:The method is simple,rapid and reliable for quality control of the capsules

Objective To study the relationship between asthma and mRNA expression of β_2AR and IL-12 gene;to determine the expression level of β_2AR, IL-12 related to CysLT_1-receptor;to analyze the correlation between the severity of asthma and the level of the genes expression. Methods White blood cells was drawn from peripheral blood mononuelear cells of asthmatic children. DNA was taken and related genes were analyzed for half quantitative analysis by D-congeal glue analyzer software. Results The level of IL-12. β_2AR mRNA expression in the asthma group is obviously lower than healthy controls (P < 0.01) ;when asthma attacks, the expression of IL-12 is positively correlated to that of β_2AR (r = 0.34, P = 0.001) and negatively correlated to that of CysLT_1-receptor (r = -0.92, P = 0.001), the expression of β_2AR is negatively correlated to the expression of CysLT_1-receptor (r = -0.85, P = 0.003). The express of IL-12 is not related to the severity of asthma (P = 0.16), while there was a negative correlation between the expression of β_2AR and the severity of asthma (P = 0.003). Conclusions The expression level of IL-12, β_22AR in the asthmatic children is obviously lower than normal;The expression level of IL-12 is positively correlated to β_2AR in PBMC of asthmatic children;The expression level of CysLT_1-receptor is negatively correlated to IL-12 and β_2AR in the PBMC of asthmatic children;The expression of β_2AR is negatively correlated to the severity of asthma.

AIM: To study the effect of bioactive compounds from Entomogenous fungi (BCEF0083), on AVP content of hypothalamus, pituitary and expression of glucocorticoid receptor (GR) mRNA of hippocampus in chronic unpredictable stress model of depression in rats. METHODS: The depression animal model was induced by chronic unpredictable stress. The effect of BCEF0083 on AVP content in hypothalamus and pituitary was tested by radioimmunoassay. RT-PCR was used to test the expression of GR mRNA in hippocampus. RESULTS: BCEF0083 decreased the AVP content of hypothalamus and pituitary in chronic unpredictable stressed rats, and increased the expression of GR mRNA of hippocampus in chronic stressed rats. CONCLUSION: BCEF0083 exhibits antidepressant effect by attenuating the stimulated function of hypothalamus-pituitary-adrenal axis in chronic unpredictable stress model of depression in rats.

OBJECTIVE:To explore the use of antibiotics in the department of respiratory diseases and pediatric outpatient department after the introduction of initiative intervention stragety on the rational use.METHODS:A field investigation was carried out on the international RDU index of antibiotics in the department of respiratory diseases and pediatric outpatient department by self-control test.Initiative intervention on physicians' prescribing practices was carried out at four stages:befter intervention,following first intervention,second intervention,and third intervention,respectively.Data statistics was analyzed by SPSS11.0 software.RESULTS:The percentages of patients treated with antibiotics and injcetable antibiotics in the department of respiratory diseases dropped from 61.6% and 19.2% before intervention to 30.0% and 14.0% after third intervention(P