BACKGROUND: How to acquire high-purity epidermal stem cells (ESCs) and maintain ESC phenotype and in vitro proliferation characteristics are urgently solved problems in the in vitro culture of ESCs.OBJECTIVE: To establish methods to in vitro isolate and culture human ESCs.DESIGN,TIME AND SETTING: Cell observation was performed in the Department of Bum,First Affiliated Hospital of Nanchang University.MATERIALS: Foreskin was obtained front 2-12-year-old patients who were subjected to circumcision.These patients did not suffer from urinary system infection.METHODS: Keratinocytes and fibroblasts were isolated from foreskin using Dispase Ⅱ and trypsin.ESCs were isolated by rapidly adhering to type Ⅳ collagen.The supematant fluid of fibroblasts of passages 2-3,which were in the exponential growth phase,was collected.After screening,the supematant fluid was mixed with serum-free medium of keratinocytes at the proportion of 1:1.Simultaneously,fetal bovine serum,epidermal growth factor,and bovine pituitary extract were added to prepare medium of ESCs,which was used for in vitro proliferation of human ESCs.Homologous keratinocytes were used as controls.MAIN OUTCOME MEASURES: After 2 passages,cloning efficiency and cloning sustaining time were calculated by plate clone formation assay.Expression levels of β 1 integrin and keratin 19 were detected by immunohistochemistry.RESULTS: The cloning efficiency of human ESCs was significantly increased,and the cloning sustaining time was significantly prolonged as compared with homologous keratinocytes cultured in the same batch (P ＜ 0.01).lmmunohistochemistry results demonstrated that both β 1 integrin and keratin 19 were positive in the human ESCs.CONCLUSION: Keratinocytes could be successfully isolated by means of rapid adherence to type Ⅳ collagen and cultured with conditioned medium in vitro.

Objective To investigate CT features of XGC in order to improve pre-operative diagnosis of the disease.Methods The CT features of 6 cases with XGC confirmed by histopathology were analyzed retrospectively.Results The gallbladder walls thick-ened in all 6 cases,diffusely thickened in 4 cases and focally thickened in 2 cases.Hypodensity nodules or bands in thickened walls were seen in all 6 cases.The continuous mucosal lines were observed in 4 cases.After contrast administration,the hypodensity band sign was seen in most cases in artery phase.Stones were found in all 6 gallbladders and in 4 common bile ducts,dilatation of biliary tract was seen in 3 cases.There were hepatic involvement in 5 cases,duodenal,hepatic flexure of the colon and antrum of the stom-ach adhesion with gallbladder was seen in 1 case respectively.Conclusion Hypodensity nodules or bands in thickened gallbladder walls and continuous mucosal line are characteristics of XGC,which are helpful in diagnosis and differential diagnosis.

Objective:To study the anti-proliferative effect of lupeol on human bladder cancer T24 cell line and the regulating mechanism for p53/miR-34a signaling. Methods:CCK-8 assay was performed to evaluate the effects of lupeol at different concentra-tions on cell viability in 24 h and 48 h. Caspase inhibitors were used to identify the subtypes of Caspase during lupeol induced cell death. The effects of lupeol on the expression of total p53 protein and miR-34a were evaluated by western blot and real-time PCR, re-spectively. The effects of lupeol on downstream targets of miR-34a were quantified by real-time PCR. Results:Lupeol could inhibit the proliferation of T24 cells in a dose-dependent manner. The IC50 of lupeol was (77. 23 ± 6. 78) μmol·L-1 in 24 h. Compared with the control group, lupeol could elevate the expressions of p53 and miR-34a (P<0. 01). Moreover, the mRNA expression of miR-34a tar-gets, Bcl-2, CD44 and c-Myc were significantly down-regulated after the treatment with lupeol (P<0. 01). Conclusion:Lupeol can inhibit T24 cell proliferation, which is related with the regulating effects on p53/miR-34a signaling.

We collected the clinical information of 21 in-patients diagnosed as Lyme disease in Peking Union College Univemity Hospital from Apr. 1994 to Jan. 2008. The clinical manifestations, laboratory tests, therapy and prognosis were analyzed retrospectively. 16 of 21 (76%) patients had fever, 14 of 21 (67%) eases had skin lesion, 17 patients had nervous symptoms, 10 cases showed arthrosis and only 2 of 21 eases involved in heart. The antibiotics of penieilin,ceftriaxone and doxyeycline were used for therapy. All of the 21 patients improved clinically and discharged. 12 cases were followed up for 1 month to 12 years and 2 of them relapsed, both of whom were in Stage Ⅲ. 8 cases remained persistent symptoms. As the manifestations of Lyme disease are complicated, for clinical borderline cases the prompt detection of etiological agent may be useful for early diagnosis. The antibiotic therapy should begin as early as possible to avoid developing chronic Lyme disease.

Objective To investigate the effect of dehydroascorbate on reactive oxygen species (ROS) in mesangial cell induced by high glucose. Methods Mesangial cells were cultured in RPMI-1640 medium containing 10% newborn calf serum. Intracellular AA and DHA contents were measured with vitamin C assay system. The intracellular formation of ROS was detected with the fluorescent probe CM-H2DCFDA by using confocal microscopy. Activity of AP-1 was detected by EMSA. Results AA entry into cells was not significantly different from background noise. At a DHA concentration of 1 mmol/L, increasing concentrations of glucose competitively inhibited DHA entry into the cells such that the accumulation of DHA was smaller than half maximal at about 22 mmol/L glucose. Cytochalasin B,a kind of hexose transporter inhibitor,inhibited DHA entry into the cells. At a glucose concentration of 25 mmol/L, DHA inhibited intracellular ROS formation in a dose-dependent manner when DHA level was smaller than 4 mmol/L. In addition, the inhibitory effect of DHA on ROS generation was accompanied by lowering AP-1 activity in mesangial cell incubated by high glucose. Conclusions Mesangial cells are DHA dependent. VitC exclusion from mesangial cells through competition of glucose and DHA for common transport mechanism will deprive the cells of the central antioxidant and can lead to ROS accumulation. Proper doses of DHA will protect mesangial cell from injury of high glucose by inhibition on ROS formation and AP-1 activation.

Objective To evaluate the effects of patient controlled analgesia(PCEA)on the perioperative changes of circulatory and pulmonary function of elderly with hypertensions after abdominal surgery.Methods Twenty-eight patients of ASAⅡ-Ⅲ aged more than 60 years undergoing uratomy were randomly divided into two groups:control group and PCEA group.Preoperative and postoperative circulatory and pulmonary functions were measured with noninvasion circulatory monitor and pocket lung function meter respectively.Results In control group,the systolic pressure,diastolic pressure,and heart rate increased by 19%,17% and 19%,respectively,as compared with preoperation.The percentage of forced vital capacity(FVC%),percentage of forced expiratory volume in first second to forced vital capacity(FEV1%) and percentage of maximal ventilatory volume(MVV%) of postoperation in control group were significantly decreased compared with preoperation(P

Objective To investigate the genetic heterogeneity of 5’-NCR of HCV genotype 1b.Methods We selected 147 HBV genotype 1b serum samples. HCV 5’-NCR fragments were amplified from these samples by use of RT-PCR assay and sequenced after using restriction endonuclease Mbo Ⅰ and BamH Ⅰ. We analyzed the phylogenetic trees of the samples and compared them with 40 isolates of HCV genotype 1b.Results The sequencing reports indicated the isolates of HCV genotype 1b recognized by BamHⅠat position 117 by a substitution (C-T) at position 120 in HCV genotype 1b viruses in China. Of 147 samples, 17 (11.56%) samples for one BamHⅠrecognization position, 26 (17.69%) for neither BamH Ⅰ recognization position nor Mbo Ⅰ,6 samples (4.08%) for two Mbo I recognization position, 18(12.24%)for one MboⅠrecognization position.Conclusions Of 147 HBV genotype 1b serum samples, 54.42% for one Mbo I recognization position, whether this have relationship with the response to IFN therapy provide the framwork for future detailed investigation of HCV antviral therapy. There is specific BamH I recognization position in isolates of HCV genotype 1b in China as compared to the other 40 HCV 1b isolates. The role of this specific mutation needs to be further researched.

Objectives In our studies before, we reported mutation C-T at -222 of hepatitis C virus type 1b genotyped on 5′ noncoding region (5′NCR). There was no this mutation of type 1b recorded in GenBank. Since genotypiog based on 5′NCR cannot differentiate subtypes, it is necessary to investigate whether the hepatitis C virus genotype 1b strains with mutation C-T belongs to other subtypes of genotype 1. Methods 64 HCV genotype 1b samples were amplified from 5′NCR by RT-PCR.Then the products were digested by use of BamHⅠ restriction enzymes. We randomly chose 6 samples with BamHⅠ restriction site and subjected them to amplification of 5′NCR and NS5B. The sequences of 5′NCR were analyzed. Sequences of NS5B fragments from the 6 samples were respectively subjected to phylogenetic analysis with subtypes of genotype 1 and 38 complete genomes of genotype 1b from GenBank.Results The phylogenetic analysis of 6 samples and subtypes of genotype 1 indicated that the genotypes of 6 strains with BamHⅠ restriction site in 5′NCR were type 1b, instead of subtype of type 1. Tree construction with 38 complete genomes of genotype 1b showed that the 6 mutants of genotype 1b did not belong to the same branch of the tree and there were no genetic differences between the mutants and the other strains of genotype 1b.Conclusions Our research indicated that the hepatitis C virus stains of C-T in 5′NCR were mutants of type 1b. Since the 5′NCR is a highly conserved region of HCV, the mutation might have some relationship to the long-time IFN therapy.

Over-expression of APP and Swedish mutation could cause some familial early onset AD. In this study, a primary screening was conducted of effective small interference RNAs (siRNAs) targeted wild type APP (APPwt) and Swedish mutant APP (APPswe). One siRNA targeting APPwt and the other siRNA targeting APPswe were designed. All these siRNAs were endogenously expressed by siRNAs expressing plasmids. COS-7 cells were transiently co-transfected with APP-GFP recombinant plasmids and siRNA expression vector. The silencing effect of each siRNA was quantitatively assessed by the level of expression of green fluorescent protein (GFP). It was found that the siRNAs silenced APPwt and APPswe to different degrees. siRNA directed against APPswe was more effective in suppressing the expression of fusion gene of APPswe than that of APPwt. The silencing effect of siRNA directed against APPswe indicating allele-specific silencing property of the siRNAs. Therefore, siRNAs directed against APP play an important role both in the therapeutic study of Alzheimer disease and functional exploration ofAPP gene.

Objective To evaluate the necessity of construction of hospital information system integration platform for county Hospital as the urban and rural medical hub in the context of the development of modern medical information.Methods By referring to the development of the domestic and international integration platform,and comprehensively considered the development status of the county hospital,and analyzed based on the perspective of sustainable development.Results The hospital information integration platform was formed to meet the needs of the county hospital,and it platform was used to open up the internal and external medical information resources,realize the integration of business resources and process optimization..Conclusion The county hospital building information system integration platform is the inevitable trend of the current construction of information technology,and its comple-ted construction can enhance province -city-county tertiary care facility data communication,reduce the coupling within the hospital business systems,and make the county hospital more flexible in the future construction and devel-opment.