Objective To establish a cell model that is close to the HCV replication in vivo and can support long-term HCV replication in vitro.Methods A human liver cell line 7721 was inoculated with serum from a chronic hepatitis C patient for 8 hours.After incubation,the presence of HCV RNA,the expression of HCV antigen and the location of HCV RNA in cells and/or supernatant were detected by RTPCR,immunohistochemistry and in situ hybridization respectively.Results It was found that plus-and minus-strand of HCV RNA could be discontinuously detected in both cells and supernatant as late as 66 days after inoculation even if cells had been subcultured for 6 times.HCV NS3、NS5 antigens could be expressed in cells and HCV RNA was mainly located within cytoplasm.Conclusion The above results suggested that 7721 cell line was not only susceptible to HCV but also could support its long-term replication in vitro.This HCV infection model in vitro was proved to be a useful tool for studying the mechanism of HCV infection and replication,as well as evaluating the antiviral compounds and screening the protective antibodies/vaccines.

Four synthetic peptides,CP10,CP9,CP33c and CP100,with the sequences corresponding to the capsid protein and nonstructural protein of hepatitis C virus(HCV)respectively,were employed to establish ELISA to detect anti-HCV.Serum HCV RNA was identified with reverse transcriptive polymerase chain reaction(RT - PCR)for comparison.Out of 320 sera collected from patients with viral hepatitis,patients with other diseases and blood donors,49 were positive for antibodies against a single or several peptides.Among the positive rates of antibodies against a single peptide,anti-CP10 rate(9.69%)was the highest and anti-CP33c(3.44%)the lowest.The positive rate for anti-HCV antibodies was significantly higher when 4 peptides were combinedly used than when only one peptide(P

AIM: To study whether there was a correlation between central foveal thickness (CFT) assessed with optical coherence tomography (OCT) and visual acuity of patient with high myopia after phacoemulsification and intraocular lens implantation.METHODS: Totally 67 patients with high myopia underwent phacoemulsification and intraocular lens implantation were enrolled in the study.Best corrected visual acuity (BCVA) was recorded and CFT was measured using OCT at 1wk,1 and 3mo after oerations.BCVA and CFT were compared before and after the operation.All patents were divided into two groups by the BCVA at 3mo after operation,BCVA>0.5 in Group A and BCVA≤0.5 in Group B.ANOVA,Spearman correlation analysis and independent t test were used for statistical analysis.RESULTS: There was a statistically significant difference between preoperative BCVA and postoperative BCVA (F=115.04,P<0.01).Preoperative CFT was different compared with that 1wk and 1mo after operation (P=0.04,0.02) and was not different with that 3mo after operation(P=0.52).There was a statistically significant difference in CFT of postoperative 3-month compared with that of postoperative 1-week(P<0.01) or that of postoperative 1-month (P<0.01).BCVA showed significant positive correlation with CFT without foveal lesion on postoperative 3mo (r=0.28,P=0.03).CFT of Group A and Group B was significantly different at 3mo after the operation (t=-2.24,P=0.03).There was no significant difference in age and intraocular lens of two groups.CONCLUSION: Optical coherence tomography allow for objective assessment of retinal construction changes in eyes with high myopia are correlated to visual acuity.

There is remarkable heterogeneity in the history of the bladder cancer,which makes it essential to evaluate individualized prognosis risk of each patient.The Nomograms model is more convenient and accurate than other prognostic models,can provide significantly accurate individualized risk estimations that facilitate management decisions.The Nomograms model has been regarded as the most accurate and discriminating prognosis model to the patient of bladder cancer,and its development has a bright future.While the Nomograms Model has several weakness itself,which would be remedied through more researches and new predictors.Before Nomograms Model can be accepted in practice,we really need better evidence that they improve patient care and outcomes.

Objective To investigate the inhibition effects of different antihistamines on the expressions of chemokines and cytokines released by nasal polyps in vitro.Methods The fresh nasal polyps resected under endoscope from the patients admitted to our hospital were cultured with mizolastine,cetirizine,loratadine,fexofenadine,dexamethasone,or ciclosporin A respectively for 24 h,with the addition of histamine and arachidonic acid.The total RNA of polyps was obtained and purified with tripure isolation reagent.The mRNA expression levels of monocyte chemoattractant protein-1(MCP-1),monocyte chemoattractant protein-3(MCP-3),regulated on activation,normal T cell expressed and secreted(RANTES),eotaxin were analyzed by RT-PCR.The concentrations of interleukin(IL)-4,IL-5 and tumor necrosis factor-?(TNF-?)in cultured supernatant was detected by ELISA.Results The MCP-1 mRNA expression in nasal polyps treated by mizolastine was weaker than those by loratadine and fexofenadine;the MCP-3 mRNA expression by mizolastine was weaker than those by cetirizine and loratadine;the RANTES mRNA expression by mizolastine was weaker than those by loratadine and fexofenadine;the eotaxin mRNA expression by mizolastine was weaker than that by cetirizine(all P

Objective To establish practical cell model of HCV infection, and investigate the susceptibility of a human liver cell line HepG2 to hepatitis C virus in vitro. Methods A human liver cell line HepG2 was incubated with serum from a chronic hepatitis C patient for 6～8 hours. Both the plus and minus strands of HCV RNA in infected cells or supernatant were examined by reverse transcription polymerase chain reaction (RT PCR). The HCV NS 3,NS 5 antigens in infected cells were respectively detected with the monoclonal antibodies to antigens of their own by immunohistochemical assay. The minus strand of HCV RNA in infected cells were localized by in situ hybridization. Results The intracellular plus or minus stands of HCV RNA were first detected on day 3 post incubation and then could be intermittently detected until day 35 post incubation in cells or supernatant. The positive signals of NS 3,NS 5 antigens could be expressed within cytoplasm of infected cells. The minus strand of HCV RNA was located within cytoplasm by in situ hybridization. Conclustions These results show that a human liver cell line HepG2 is not only susceptible to HCV but also able to support its long time replication in vitro.

Objective To explore the expression of caspase-3 and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors in the CD4+ and CD8+ T cells of systemic lupus erythematosus (SLE)patients. Methods The CD4+ and CD8+T cells from peripheral blood of 20 SLE patients and 10 healthy volunteers were separated using magnetic cell sorting system (MACS), reverse transcription-polymerase chain reaction (RT-PCR) was used to test the expression of caspase-3 and TRAIL receptors in CD4+ and CD8+T cells of SLE patients and healthy volunteers. Results The CD8+T cells from SLE patients had significantly higher caspase-3 (P=0.003) and TRAIL-R2 (P=0.024) expression than those from healthy volunteers. Conclusion The TRAIL-R2 signal pathway may be a possible important pathway to the apoptosis of T cells in SLE patients.

Objective To clone, express and purificate human MRPS17 cDNA in Escherichia coli (E. coli). Methods MRPS17 cDNA was obtained from total RNA isolated from primary cultured human hair papillary cells by RT-PCR and sequencing method, and then it was inserted into prokaryotic expression vector pET28a for the IPTG-induced expression in E. coli BL21 (DE3). The expression product fused with 6?His at C-terminal was analyzed by Western blotting, and purified by using Ni 2+-NTA ion exchange resin. The purity of MRPS17 protein was analyzed by SDS-PAGE. Results Human MRPS17 cDNA was obtained, and the expression plasmid pET28a-MRPS17 was constructed successfully. Western blot analysis showed that human MRPS17 with 13kD molecular weight was expressed in E. coli at 20℃. The purity of the recombinant MRPS17 was more than 90% after purification using Ni 2+-2NTA ion exchange resin. Conclusion The sequence of MRPS17 cDNA was consistent with the known sequence from the Genbank. MRPS17 is successfully induced and expressed in E. coli.

Humans ; Infant ; Male ; Mycoses ; pathology ; Penicillium ; isolation & purification

Objective To investigate the expression of T cell factor-4 (TCF4) gene in dermal papilla cells of hair follicles.Methods The expression of TCF4 gene was examined by in situ hybridization in scalp tissues of patients with alopecia areata and normal human controls,The protein and mRNA exprcssions of TCF4 were detected by immunochemistry and RT-PCR method,respectively,in aggregated and non-aggregated human dermal papilla cells.ResultsAs shown by in situ hybridization,TCF4 gene was expressed in the dermal papilla cells from healthy controls,but not in those from patients with alopecia areata.Both cell immunochemistry and RT-PCR showed that TCF4 gene expressed in aggregated dermal papilla cells,but not in non-aggregated dermal papilla cells.ConclusionsTCF4 gene is expressed in dermal papilla cells.The growth cycle Of follicles may be related to wnt signal.