Objective To investigate the differentiation potenitial of ES cell derived epidermal like stem cells,to lay a base for the study of their differentiation mechanism,as well as seek new source to provide seed cells for skin engineering. Methods Mouse ES cells labeled or unlabeled by Hoechst 33342 were cocultrued with human amnion for 4 days.The epidermal like stem cell clones formed on the surface of amnion were digested with trypsin and transplanted into hypodermis of nude mice for 10,20 and 45 days,then the differentiation pattern of the donor cells were observed and estimated with morphological and immunohistochemical method. Results The grafts may differentiate into tubular or follicular like structures lined with simple or stratified epithelial like cells which expressed ? 1 integin,CK19,CK15,PK involucrin and CEA respectively after 10 to 30 days of transplantation.Keratinized stratified squamous epithelium,sebaceous gland like,sweat gland like and hair follicle like structures were observed after 45 days ofter transplantation.Conclusion ES cell derived epidermal like cells might have differentation potential to diffreentiate into keratinized stratified squamous epithelium,sebaceous like,sweat gland like and hair follicle like structures.

AIM: To investigate the mechanism by which human amnion induced mouse embryonic stem (ES) cells to differentiate into epidermal like cells. METHODS: ES-BALB/c cells were cocultured with human amnion in transwells for 4-5 days, and those cultured alone without amnion were taken as control group. The morphological differentiation were observed. The committed differentiation of ES cells into epidermal like cells were detected by integrin-?_1, CK19, CK15 and involucrin immunohistochemistry, respectively. RESULTS: After 4-5 days of coculture, ES cells differentiated into single layer of epidermal like cells, fitted tightly, with polyhedral in shape. The immunohistochemical staining results showed that, most of the cells were integin-?_1 positive, only a few cells were CK19 and CK15 positively stained. Most of the cells in control group died, the survived ones were different in morphological shapes, and no integrin-?_1, CK19 and CK15 positive cells were found. CONCLUSION: Soluble substances secreted by human amnion may play an important role in inducing the differentiation of mouse ES cells into epidermal like cells. [

Objective To investigate water treatment technology and pollution status of Giardia lamblia and Cryptosporidium parvum in resource water and finished water of the centralized water supply enterprises in Shenzhen,and to provide scientific basis for waterborn infectious disease control.Methods Field hygiene assessment were conducted on the resource water and water treatment technology of 19 water supply enterprises during May to July,2008.One water sample was collected from resource water and finished water in each centralized water supply enterprise respectively,and was detected for Giardia lamblia and Cryptosporidium parvum.Results The routine traditional water treatment technology including coagulation deposition,quartz sand filtration and chlorine disinfection was applied in all of the 19 centralized water supply enterprises.Giardia lamblia and Cryptosporidium parvum were not found in samples from the source water and finished water.Conclusion The centralized water supply system in Shenzhen isn't polluted by Giardia lamblia and Cryptosporidium parvum.

The significant differences in prevalence rate of essential hypertension among the races in Xinjiang, i.e. in Kazakh, Han, and Uighur had been shown in 1979 screening. They were 6.6%, 3.5% and 2.6% respectively.The present investigation shows that some dietary constituents as well as the nutritional imbalance was involved in the causal effect in the development of hypertension.Three race people living in three quite different areas (geographic, climatic environmental conditions etc,) were investigated in April to July 1981. Blood pressures of 9,313 persons were measured. The dietary pattern and staple food items were studied for 491 persons within 69 families.The findings suggested that: l)the prevalence rate of hypertension was identical to the previous screening, 2) there was a positive correlation between the high consumption of protien (r= 0.974), fat(r = 0.622) and the prevalence rate of this disease, but a negative correlation in vitamin C intake (r = -0.620).The excessive intake of salt was accounted as a risk factor for poss- ible association with this disease only in Kazakh people who were living in various areas.

Objective To investigate the condition which can induce mouse ES cells to differentiate into epidermal-like stem cells for the clinical application of ES cells-derived epidermal-like stem cells and the research on the mechanism of committed differentiation of ES cells. Methods Coculture mouse ES cells with human amnion for 3-4 days, and the committed differentiation were detected by flow cytometry and immunohistochemistry. In experimental group 1, amnion was pasted covering the whole bottom of the wells, with the epithelial surface upward, and in group 2 covering the half bottom. No amnion was used in control. Results After 3-4 days of co-culture, epidermal-like stem cell clones were formed on the epithelial surface of amnion in group 1 and group 2, and expressed high levels of integrin-? 1, CK19 and CK15. The percentages of integrin-? 1, CK19 and CK15 positive cells counted in group 1 by flow cytometry were 89.2%, 86.8% and 71.2% respectively, versus the control group of 8.4%,9.6% and 11.8%, the differences were significant in all the three indices (Z tests P

Objective To compare the expression level of calpain and calpastatin mRNA in rheumatoid arthritis (RA) and osteoarthritis (OA) synovial tissues (ST) and analyze the antigen epitopes of human calpastatin recognized by RA sera.Methods Total RNA of three RA and two OA patients was isolated from ST and expression levels of calpain and calpastatin mRNA were detected by reverse transcription polymerase chain reaction (RT PCR).According to hydrophilicity of human calpastatin (Accession,U 31346), overlapping oligopeptides were synthesized by auto spot Robot in N terminal (L domain) and C terminal (Ⅳ domain) of calpastatin.The epitopes of synthesized peptides were analyzed by RA sera using dot ELISA.Results mRNA expression of calpain showed higher than that of calpastatin in both OA and RA.Meanwhile,the mRNA expression level of calpain increased in RA more than that in OA.Three antigen epitopes of human calpastatin (with the sequence DKDLDDALD,DTIPPEYRH and QDPIDALSG) were identified by RA sera,while control sera failed to react with synthetic peptides.Conclusion The calpain calpastatin system may participate in pathogenic mechanism of RA,and the calpastatin may be a target for autoantibody of RA,which implies that it is not only possible to treat RA patients with synthetic peptide of human calpastatin,as a chemotherapeutant,but also a materials for immunodiagnosis of RA.

Objective To explore the efficacy and protective mechanism of calpain, a calcium activated neutral proteinase as a vaccine candidate molecule. Methods Anti calpain sera were prepared by immunized BALB/c mice with recombinant calpain. Anti calpain sera, schistosomula of Schistosoma japonicum , and mouse eosinophils were incubated at 37 ?C , 5% CO 2 for 48 hours, and the adherence of eosinophils to schistosomula and its schistosomulacidal efficacy were observed. Results Mice immunized with recombinant calpain produced a high level IgG antibodies specific to the antigen immunized. Immunoblotting analysis showed murine anti recombinant calpain sera bound specifically to recombinant calpain. Ninety six percent of schistosomula were surrounded by cells when incubated with mouse eosinophils, and a significant number of dead schistosomula was observed at 48 hours when incubated with mouse serum and eosinophils as compared with control serum and eosinophils ( P

Objective To investigate the effect of recombinant human interferon α-2b on influenza virus in vitro.Methods Influenza A virus subtype H1N1 and influenza B/Y virus were inoculated into Vero cells and different concentrations of interferon α-2b and oseltamivir were added.Numbers of virus plaques were observed and calculated,and quantitative RT-PCR were used to assess the inhibitory effect of interferon α-2b and oseltamivir in vitro.The nuclear export of viral ribonucleoprotein (RNP) complexes were monitored under fluorescence microscope.Results Virus plaque test showed that influenza A viruses subtype H1N1 were significantly inhibited when 10 μg/μL interferon α-2b and 10 μg/μL oseltamivir were added,and the numbers of plaques were 7.5 × 108 and 15 × 108 PFU/mL,respectively;the inhibitory effect of oseltamivir was better than that of interferon α-2b.Influenza B/Y viruses were also inhibited when 10 μg/μL interferon α-2b and 10 μg/μL oseltamivir were added,and the numbers of plaques were 1.1 × 108 and 1.5 × 108 PFU/mL,respectively.Quantitative RT-PCR results showed that the cycle threshold (CT) values of influenza A virus subtype H1N1 and influenza B/Y virus were much higher when 10 μmol/L interferon α-2b and 10 μmol/L oseltamivir were added.CT values of influenza A virus subtype H1N1 were 16,26 and 35 before and after inferferon α-2b and oseltamivir were added.CT values of influenza B/Y virus were 18,27 and 31 before and after interferon α-2b and oseltamivir were added.Reduction in the nuclear export of viral RNP in influenza A virus subtype H1N1-infected Vero cells was also observed when 10 μmol/L interferon α-2b were added.Conclusion Interferon α-2b has significantly inhibitory effect on both influenza A virus subtype H1N1 and influenza B/Y virus in vitro.

Objective:To study the effect ofmetformin combined with insulin aspart on the serum cholesterol(TC),total Bilirubin (TBil),uric Acid(UA),urinary Micro Protein(mAlb) levels and Maternal and Infant Outcomes of gravida with Gestational Diabetes Mellitus.Methods:84 patients ofgestational diabetes mellitus who received therapy from June 2014 to June 2016 in our hospital were selected.According to random number table,those patients were divided into the observation group (n=42) and the control group (n=42),on the basis of routine treatment,The control group was treated with insulin aspart,while the observation group was combined with metformin hydrochloride.The blood glucose index and the levels of TC,TBil,UA,mAlb and maternal and infant outcomes were compared.Results:After treatment,the levels of fasting blood glucose (FBG),postprandial 2h blood glucose (2hPG),glycosylated hemoglobin (HbA1c),TC,TBil,UA and mAlb in the observation group were significantly lower than the control group,the levels of TBil was significantly higher than the control group (P＜0.05);the incidence ofgestational hypertension,hydramnios,premature birth,cesarean section,giant child and neonatal jaundice were significantly lower than the control group (P ＜0.05).Conclusion:Metformin combined with insulin aspart was well for gestational diabetes mellitus,which could effectively improve the blood glucose indicators and TC,TBil,UA,mAlb levels,maternal and infant outcomes.