Objective To investigate the functional role of mur in e hepatocyte growth factor (HGF) in inducing in vitro the mouse embryonic st em cells (ESC) to differentiate unidirectionally into hepatocyte. Method s Both spontaneous and HGF induced differentiations of the primordial c ell elusters formed by suspended culture in vitro of ESC were daily observed with microscope for 5-7 days. After 4 weeks of culture, the cells were stained with HE and glycogen staining, their morphology were observed, The synthesis of urea nitrogen and triglycerides in the culture medium, the expression of myocard ial MHC, albumin, AFP and CK18, and the indocyanine green (ICG) staining were a lso detected. Results It was hard to control the spontaneous di fferentiation of ESC. HGF could promote the differentiation of ESC into endoderm , and more likely into myocardium. HGF could also induce the expression o f albumin, AFP and CK18, and positive staining of ICG and FDA. Conclusio ns HGF may induce the differentiation of ESC into hepatocyte, but the f unctional role is limited. It implies that a comprehensive effort of multiple fa ctors might be needed in inducing the hepatocyte differentiation.

Objective To establish a LC-MS analytical method for determination of N,N-dimethylaniline which is genotoxic impurity in quetiapine fumarate.Methods The method was achieved by an Waters ACQUITY UPLC CSHTM PhenylHexyl(2.1 mm× 100 mm,1.7 μm) utilizing a mobile phase of buffer-methanol(900∶ 100) (A)-acetonitrile(B) with gradient elution at the flow rate of 0.4 mL·min-1.The temperature of column was set at 50 ℃;The DIONEX Ultimate 3000 HPLC-AB Science 4000 QTrap Tripling Four bar LC-MS to detect N,N-Dimethylaniline (ESI source,in MRM positive mode).Results Standard curve was linear in the range of 0.4-8.0 ng(r =0.999 3);The limit of detection was 0.2 ng;The limit of quantification of N,N-dimethylaniline was 0.4 ng,respectively.The average recovery of N,N-dimethylaniline was 103.3 ％;RSD was 4.3％ (n =9),respectively.Conclusion The method is convenient and sensitive for the determination of N,N-dimethylaniline in quetiapine fumarate.

AIM: To establish the quality control for Lidanzhitong Tablet (Radix Paeoniae Rubra,Fructus Aurantii,Rhizoma Corydalis,etc.). METHODS:Radix Paeoniae Rubra, Fructus Aurantii and Rhizoma Corydalis were identified by TLC, and the content of paeoniflorin in Radix Paeoniae Rubra was determined by HPLC. RESULTS: Paeoniflorin showed a good linear relationship at a range of 0.597-4.997 ?g, r=0.999 9. The average recovery was 99.06%, and RSD was 3.4%. CONCLUSION: The method is accurate and reliable, and can be used to control the quality of this preparation effectively。

Objective The purpose of this study was to investigate the effects of gossypol acetic acid (GAA) on protein and mRNA expressions of hMLH1 gene in human tongue carcinoma cell line Tca8113 in vitro in order to discuss the mechanism of tumor suppression of GAA. Methods (1) Western-blot was used to study the effects of GAA on protein expressions of hMLH1 gene in Tca8113 cell line treated by different concentrations of GAA for 48 h. (2) Real-time fluorescence quantitative PCR (RFQ-PCR) was used to investigate the effects on the mRNA expressions of hMLH1 gene in Tca8113 cell line treated by GAA for 48 h. Results (1) Compared with the control group, the results of Western-blot showed that the protein expression of hMLH1 gene was increased after treatment by GAA for 48 h ( <0.05) . (2) The results of RFQ-PCR indicated that the mRNA expression of hMLH1 gene was increased after GAA treatment for 48 h ( <0.05) . Conclusion GAA could up-regulate protein and mRNA expression of hMLH1 in Tca8113 cell line, which indicated that it may be one of the mechanisms of tumor suppression effect of GAA.

Objective:To study the effects of gossypol acetic acid(GAA)on the methylation level and the mRNA expression of hM-LH1 in human tongue cancer Tca8113 cells.Methods:Tca8113 cells were treated by GAA at various doses for 24 h,48 h and 72 h respectively.MTT assay was used to detect the cell proliferation.Nested methylation specific PCR(nMSP)was used to detect methyl-ation level of hMLH1 .Real-time fluorescence quantitative PCR(RFQ-PCR)was applied to investigate the mRNA expression of hM-LH1 gene.Results:GAA inhibited the proliferation of Tca8113 cells dose-and-time dependently,decreased the DNA methylation level of hMLH1(P<0.05)and increased hMLH1 mRNA expression in Tca8113 cells(P<0.05).Conclusion:GAA can suppress proliferation of Tca8113 cells by demethylation of hMLH1 gene and increase of hMLH1 mRNA expression.

Objective:To explore molecule mechanism about anti-aging of bifidobacterium by investigating the effect of lipoteichoic acid (LTA) of bifidobacterium on thymus of aging mice induced by D-galactose.Methods:Male Kunming mice were divided into different groups at random. These mice were induced aging by subcutaneous injection of D-galactose on the back of mice and LTA was simultaneously administered to them daily for 40 days. Body weight, thymus index ,thymus morphology, thymus c-fos, p16 production and peripheral blood IL-2 concentration were tested by HE staining, immunochemistry method, ELISA and other methods. Results:There were significant differences for body weight ,thymus index, thymus morphology,thymus p16, c-fos production and peripheral blood IL-2 concentration between the young control group and the aging model group. Whereas, LTA can improve thymus morphology ,increase body weight ,thymus index and c-fos production, meamwhile, decrease thymus p16 and peripheral blood IL-2 concentration of aging model mice. Conclusion:The results showed that LTA can remarkably delay thymus aging, and it may be a molecular mechanism of bifidobacterium delaying immunosenescent.

Objective To investigate the changes of blood lipid,endothelin(ET)and carbohydrate tolerance in simple obesity children.Methods Serum total cholesterol(TC),triglyeride(TG),low density lipoprotein(LDL),endothelin(ET) and oral glucose tolerance test(OGTT) of 436 cases(light obesity group 242,moderate obesity group 138,heavy obesity group 56) were detected respectively.Results The levels of TC,TG,LDL,ET and OGTT in 3 simple obesity groups were all higher than those in control group obviously and there were significant differences.The significant positive correlation between the obesity children serum lipids,ET and obese degrees were observed.Four insulin resistant children were found in heavy obesity group and none in other groups.Conclusions The levels of serum lipids and ET increases in obesity children and the OGTT decreases in some heavy obesity children.For preventing the obesity related health problems,the simple obesity children should be paid more attention to earlier.

Objective To investigate the epidemic characteristics of mycoplasmal pneumonia outbreak in a middle school, and to provide suggestions for outbreak prevention and control.Methods Principle and method of field epidemiology was used and,speciously,questionnaire investigation was carried out for the outbreak of mycoplasmal pneumonia.Outbreak characteristics and corresponding emergency intervention were discussed.Results There was an outbreak of mycoplasmal pneumonia in 2 classes of the middle school,and the attack rate was 42.50%.The attack rate of other classes was 6.75%.Timely identification and diagnosis of the disease,and comprehensive control to prevent the spread of the epidemic,brought the disease under control.Conclusion The outbreak of mycoplasmal pneumoniae infection is associated with the close contact in daily activities.Emergency intervention can prevent the further spread of the disease.

Objective To explore the effect of hypoxia inducible factor-1α silencing by siRNA on proliferation, apoptosis and necrosis of human renal proximal epithelial cell ( HK-2 )under hypoxia. Methods CoCl2 was used to mimic hypoxia, and siRNA technique was used to silence HIF-1α. HK-2 cells were divided into five groups, including normal culture group,hypoxia culture group, transfection reagent group, negative control group and HIF-1α siRNA group.MTT assay was used to detect the growth inhibition ratio of cells. TUNEL and caspase-3 quantity was used to detect apoptosis. LDH activity in supernatant was detected to evaluate cell necrosis.Real-time PCR and Western blotting were used to detect mRNA and protein of HIF-1α, HIF-2α,glucose transporter 1(Glut-1) and vascular endothelial growth factor (VEGF). Results (1) The transfection efficiency of siRNA was 95%-100%. Under normoxia, the efficiency of siRNA silencing HIF-1α gene reached to 70%, while under hypoxia, it was 97%. (2) The growth inhibition ratio of cells in HIF-1α siRNA group was significantly higher than that of other groups including hypoxia culture group, transfection reagent group and negative control group (all P＜0.05). No significant difference was found in apoptotic ratio of the other four groups except normal culture group (P＞0.05). The LDH level in HIF-1α siRNA group was significantly higher than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). (3) The expression of HIF1α, Glut-1, VEGF mRNA and protein in HIF-1α siRNA group was significantly lower than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). No significant difference was observed on the level of HIF-2α mRNA and protein in the other four groups except normal culture group (P＞0.05). Conclusion HIF-1α gene silencing can inhibit the mRNA and protein expression of Glut-1 and VEGF, and can aggravate growth inhibition and necrosis of HK-2 cells under hypoxia.

Adult ; Granuloma, Respiratory Tract ; microbiology ; Humans ; Male ; Mycoses ; diagnosis ; therapy ; Pharyngeal Diseases ; diagnosis ; microbiology ; therapy