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.

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

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

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.

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.

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 screen and identify proteins interacting with hematopoietic stem/progenitor cell differentiation-related gene HSPC016, and to explore the molecular mechanism involved in the regulation by HSPC016 on the aggregative behavior of dermal papilla cells. Methods By using yeast two-hybridization,HSPC016 gene was sub-cloned into pGBKT7 to construct the bait plasmid (named as pGBKT7-HSPC016) in yeast AH109. The cDNA yeast expression library of human dermal papillae cells in yeast Y187 was screened with the bait plasmid and the proteins interacting with HSPC016 were identified. Yeast two-hybridization retransformation experiment was conducted to exclude the false positive clones and verify the interactions, then,the positive clones were sequenced and analyzed by using bioinformatic methods. Results The bait plasmid pGBKT7-HSPC016 was constructed successfully and there was no self-activation in or toxicity against yeast AH 109. Four proteins,including forkhead family of transcription factors (FOXO 1 ), mitogen-activated protein kinase 11 (MAPK 11 ), phosphoinositide-3-kinase (PIK3R3) and liver X receptor were screened and identified. Bioinformatic analysis revealed that these proteins had close relationship with intracellular energy metabolism and translational regulation. Conclusions HSPC016 may regulate the aggregative behavior of DPCs by regulating the levels of intracellular reactive oxygen species (ROS) and interacting with signaling molecules involved in intracellular energy metabolism and translational regulation.

Objective To study the protective effects of rosiglitazone against early skin changes in experimental diabetic rats. Methods Diabetic models were established in male Wistar rats aged 8 to 10 weeks by using streptozotocin (STZ). Then, 39 experimental diabetic rats were equally divided into insulin-treated diabetic group (DI group), rosiglitazone-treated diabetic group (DR group), diabetic control group (DC group),and 13 normal rats served as the control (C group). The rats were given subcutaneous insulin (1 - 2 U) twice daily in DI group, intragastric rosiglitazone (5 mg/kg) once daily in DR group, and intragastric sterile water in DC and C groups. Sixteen weeks later, heart blood samples were collected from all the rats for the measurement of tumor necrosis factor (TNF)-α, interleukin (IL)-6, C reactive protein (CRP), superoxide dismutase (SOD) and P substance (SP) levels, then the rats were killed and tissue samples were obtained from the medial area of the dorsal skin and subjected to pathological observation, measurements of skin as well as dermal thickness, and immunohistochemical examinations for the expression of advanced glycation end products(AGEs) and peroxisome proliferator activated receptor-γ (PPAR-γ). Results The levels of IL-6, TNF-α and CRP in the DC group were significantly higher than those in the C group (135.05 ± 43.39 ng/L vs. 99.92 ±32.36 ng/L, 1.45 ± 0.67 μg/L vs. 0.86 ± 0.60 pg/L, 3.51 ± 0.62 mg/L vs. 2.54 ± 1.31 mg/L, all P < 0.05),while no significant difference was found between C group and DI group or DR group (all P> 0.05). Decreased levels of SOD and SP were noted in the DC group (70.71 ± 37.52 U/ml, 22.22 ± 7.93 ng/L), compared withthe C group (137.76 ± 27.6 U/mL, 29.57 ± 3.74 ng/L, both P< 0.01), DI group (149.96 ± 13.25 U/mL, P<0.01; 29.79 ± 5.21 ng/L, P< 0.05) and DR group (128.50 ± 38.27 U/mL, P< 0.01; 33.35 ± 15.0 ng/L, P<0.05 ). Micrometer measurements indicated that the skin thickness was significantly lower in the DC group than in the C group, DI group and DR group (0.77 ± 0.18 mm vs. 1.59 ± 0.26 mm, 1.47 ± 0.50 mm and 1.22 ±0.47 mm, P < 0.01, 0.01, 0.05 respectively). Histological observation found a mild disarrangement of epidermal cells, shrinkage, swelling and degeneration of dermal collagen as well as progressive atrophy or disappearance of subcutaneous fat in the DC group. No obvious histological changes appeared in the DI or DR group. Immunohistochemistry indicated that AGEs and PPAR-γ proteins, which were stained into brown granules, were located in the vascular basement membrane, stromal cells, etc, of skin. The DC group showed the highest expression of AGEs but lowest expression of PPAR-γ. Conclusions There is an accumulation of AGEs and elevated expressions of inflammatory factors in the skin of experimental diabetic rats, and rosigLItazone shows a protective effect against the early skin changes.

Objective To investigate the genotype and antifungal susceptibility of AspergiIlus species isolated from clinical settings and environmental sources. Methods Random amplification of polymorphic DNA (RAPD) analysis was performed to profile the genotypes of 48 Aspergillus fumigatus strains, 59 Aspergillus flavus strains and 30 Aspergillus niger strains isolated from clinical settings and environmental sources. NCCLS M38-A protocol was carried out for antifungal susceptibility testing of these Aspergillus isolates. Results As RAPD analysis showed, the 48 Aspergillus fumigatus strains were classified into 8 genotypes, 59 Aspergillus flavus strains into 12 genotypes, and 30 Aspergillus niger strains into 5 genotypes. A significant difference was observed in the susceptibility to terbinafine among the 8 genotypes of Aspergillus fumigatus (χ2 = 33.092, P ＜0.01 ) as well as in that to amphotericin B among the 5 genotypes of Aspergillus niger (χ2 = 15.185, P＜ 0.05).No statistical difference was found in the susceptibility to amphoteriein B, itraconazole, fluconazole or flucytosine among the 8 genotypes of Aspergillus fumigatus or in that to terbinafine, itraconazole, fluconazole or flucytosine among the 5 genotypes of Aspergillus niger. Conclusion There is some difference in the susceptibility to some antifungal agents between different genotypes of Aspergillus fumigatus and Aspergillus niger.

Objective To observe the histopathological features of early skin lesions and serum inflammatory mediators changes in the diabetes mellitus(DM) rats.Methods A total of 50 Wistar rats at 8 to 10 weeks of age were divided into normal control group(C group,n=25) and diabetic group(DC group,n=25).The rats of DC group were given a high-glucose and high-fat diet for 2 weeks to induce insulin resistance and then an intraperitoneal injection of 35 mg/kg streptozotocin.All 25 rats were identified to be diabetic models with their fasting blood glucose over ≥7.0 mmol/L.Four rats of C group or DC group were killed to take the skin tissues in the 4th,8th and 12th weekend.In the 16th week,the heart blood samples of the left 13 rats of C group and 10 rats of DC group were collected to measure TNF-?,IL-6 and C-RP levels,and the skin samples were also taken for pathological observation and full-thickness and dermal thickness measurement.Results In the 16th week,the TNF-?,IL-6 and C-RP levels of DC group were significantly higher than the normal group(P