Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.

A pilot-scale research on purification of odorous gas emitted from wastewater treatment plant using a biofilter was conducted. The aim of this study is to check on the performance of biofilter running in various conditions and the effect of pH fluctuations on the performance of biofilter. The relation between distribution of microorganism and removal of odorous gases were also discussed here. The experimental results show that the predominant odor-causing gas can be efficiently eliminated by a biofilter inoculated with deodoring microorganism which were isolated previously. Moreover the biofilter had been proved having good tolerance to shocking loads of pollutant and can operate well in the condition of low pH.

In order to explore the dormancy physiological and biochemical mechanism of Paris seeds, the seed embryo growth courses, and the dynamic change of 5 enzymes, include SOD, POD, CAT, MDH, G-6-PDH were measured during variable temperature stratification. The results indicated that Paris seeds embryo grew quickly after 40 d in warm-stratification (18 ± 1) °C, at the meantime the metabolic activity was significantly strengthened. These facts showed that Paris seeds turned into physiological after-ripening process. After 60-80 d, the morphological embryo after-ripping process basically completed, and the following cold-stratification (4 ± 1) °C furthered Paris seed to finish physiological after-ripening. After 40 d, the activity of MDH decreased while G-6-PDH increased significantly. This showed that the main respiratory pathway of seed changed from TCA to PPP, which benifited breaking seed dormancy. In the whole period of stratification process, the activity variation of SOD and CAT was insignificantly and the activity of POD was enhanced significantly after shifting the seed in cold stratification process. This showed that SOD, CAT had no direct effects on breaking Paris seed dormancy but keeping the seed vigor, while the POD might involve in the process of Paris seed dormancy breaking.
Germination ; Liliaceae ; chemistry ; embryology ; enzymology ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; Temperature

OBJECTIVETo investigate the efficacy of PEG-interferon alpha (PEG-IFN alpha) treatment of HBeAg-positive chronic hepatitis B and HBV genotypes and liver tissues effect of HBeAg seroconversion.

METHODS54 cases confirmed by liver biopsy, genotype clear HBeAg positive chronic hepatitis B (CHB) patients according to body weight, respectively, subcutaneous injection of PEG-IFN-alpha2a 135 microg or 180 microg, or PEG-IFN-alpha2b 50 microg, 80 microg or 100 microg once weekly treatment for 48 weeks and followed for 24 weeks after discontinuation. Statistics of HBeAg seroconvertion, HBV genoty pes and liver histology e antigen seroconversion after the end of treatment.

RESULTS54 patients were followed up at the end of HBeAg seroconversion rate was 29.63% (16/54). Genotype B patients with HBeAg seroconversion rate was 35.29%, 27.03% higher than the C-type patients, but the difference was not statistically significant (chi2 = 0.382, P = 0.537). Inflammation of the liver activity highter ( > G2) , the degree of fibrosis heavier ( > S1) HBeAg seroconversion rate (50.00% vs. 25.00%, 40.90% vs. 21.88%), but were not statistically significant (chi2 = 1.391, 1.444, P = 0.238, 0.229). Activity of HBV genotype, liver inflammation, liver fibrosis and other factors by multivariate Logistic regression analysis, only liver inflammation activity of the important factors of HBeAg seroconversion.

CONCLUSIONImportant factors, liver inflammation activity of PEG-interferon alpha treatment of HBeAg-position chronic hepatitis B patients and HBV genotypes and liver fibrosis may be of little significance.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Genotype ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; pathology ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Liver ; pathology ; Logistic Models ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use

Objective To observe the dynamic changes in mesenteric lymph microcirculation and its kinetics in rats suffering from severe heatstroke (SHS), and to explore the role of mesenteric lymph in the pathogenesis of SHS. Methods SHS rat models were reproduced in an incubator with high temperature and high humidity. The vital signs and the time of onset of SHS in rats were recorded continuously during the process of heat stress. Parameters of mesenteric lymph microcirculation including Index-I, Index-II, L.D-Index, and intra-lymphatic pressure before heat exposure, 60min after heat exposure, and onset of SHS were collected and analyzed. Mesenteric lymph was collected at 30-min interval, and its volume of production was measured dynamically. Results Rat SHS model was reproduced successfully. After exposure to the environment with high temperature and high humidity, the core temperatures of the rats raised to 42°C at the time point of 60min, and HS onset occurred at about 77min. Mesenteric lymph-vessel contraction indices including Index-I, Index-II, L.D-Index, lymph-vessel pressure and mesenteric lymph flow decreased significantly at the time point of 60min (P<0.05). However, all the above parameters increased at the time point of SHS onset (P<0.05), but had not yet reached the normal levels before hyperthermia and high humidity exposure (P<0.05). Conclusion Changes of mesenteric lymph microcirculation in rats with SHS shows a dynamical regularity, which may take part in the pathogenesis of SHS.

Objective To explore the effect of mesenteric lymph from rats suffering from heat stroke (SHS) on the inflammatory activity of vascular endothelial cells. Methods Rats were placed in a prewarmed incubator to reproduce heat stroke. Human umbilical vein endothelial cells (HUVECs ECV-340) were incubated in 5% mesenteric lymph collected pre-, during-, and post-HS for 3 and 6 h respectively in vitro. The levels of high mobility group protein B1 (HMGB1), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 of ECV-340 cells were determined. The expression of intercellular adhesion molecule 1 (ICAM-1) mRNA and NF-κB activity of ECV-340 were also assayed. Results The levels of HMGB1, TNF-α, IL-1β and IL-6 induced by SHS mesenteric lymph of rats significantly increased after heat treatment (P<0.05), and this effect was enhanced with prolongation of exposure time (P<0.05). The expression of ICAM-1 mRNA and NF-κB activity were significantly elevated after being activated by SHS mesenteric lymph (P<0.05). Conclusion HS mesenteric lymph probably activates the inflammatory responses of vascular endothelium, which is closely associated with the pathogenesis of SHS.

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.

Studies concerning the association between arsenic exposure and hepatitis B virus (HBV) infection have been lacking.The present study aimed to examine the association between total urinary arsenic (TUA) and infection of HBV.A total of 5186 participants from National Health and Nutrition Examination Survey (NHANES) 2003-2014 were included in the analysis.We used logistic regression to evaluate the association.We defined two measures of TUA.TUA1 was the sum of arsenous acid,arsenicacid,monomethylarsonic acid and dimethylarsenic acid.TUA2 was defined as TUA minus arsenobetaine and arsenocholine.The results showed that the weighted overall prevalence of HBV infection was 6.08％.For NHANES 2003-2014,the medians (interquartile range) of TUA1 and TUA2 were 5.60 μg/L (3.97-8.09 μg/L) and 4.91 μg/L (2.36-9.11 μg/L),respectively.Comparing the highest quartile to the lowest quartile after multivariable adjustment showed that the odds ratios (ORs) and 95％ confidence intervals (CIs) for TUA1 and TUA2 were 2.44 (1.40-4.27) and 2.84 (1.60-5.05),respectively.In conclusion,elevated urinary arsenic was associated with the risk of HBV infection.Further studies,especially prospective studies,are needed to confirm the causal relationship between arsenic exposure and HBV infection.

OBJECTIVETo investigate the association between oral neoplasm genetic susceptibility and genetic polymorphism of p53 intron 7.

METHODSThe intron 7 ApaI polymorphism of p53 was analyzed in 95 oral neoplasm patients and 105 healthy individuals by utilizing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotyping assay technique, and direct sequencing was performed in 30 cases which were selected from the patients and controls by random sampling.

RESULTSIn oral neoplasms cases, haplotype combinations were T-G 43.2%, C-T 56.8%, and frequencies of genotype were T-G/T-G 15.8%, C-T/T-G 54.7%, C-T/C-T 29.5%, while in controls they were T-G 30.9%, C-T 69.1% and T-G/T-G 10.5%, C-T/T-G 41.0%, C-T/C-T 48.5%. There was a significant difference in the allelic frequency and the genotypical distributions between the oral neoplasm patients and the controls. The individuals with the T-G allele had a slight increasing neoplasm risk than individuals with C-T allele; the OR for T-G versus C-T was 1.69 (95% CI, 1.12 - 2.51). The risk of suffering from oral neoplasms was higher in the individuals of T-G/T-G genotype and of T-G/C-T genotype than in individuals of C-T/CT genotype with odds ratio of 2.48 versus 2.20.

CONCLUSIONSThere are two polymorphic points in the 7th intron of human p53 gene, which could be associated with genetic susceptibility of oral neoplasms. T-G allele may be the risk factor of oral neoplasms.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Introns ; Male ; Middle Aged ; Mouth Neoplasms ; genetics ; Polymorphism, Genetic ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo compare two rat models of myocardial infarction established by coronary artery ligation and harmonic scalpel method for their advantages, disadvantages and applications.

METHODSSixty Sprague Dawley rats were randomly divided into sham-operated group, coronary artery ligation group and harmonic scalpel group. In the latter two groups, myocardial infarction was induced by ligation of the left anterior descending coronary artery and coagulation of the artery, respectively. Four weeks after the operations, the rats were sacrificed to measure the serum levels of CK, CK-MB, LDH and AST, and the hearts were collected for histopathological observation with HE staining.

RESULTSThe rats in the sham-operated group and coronary artery ligation group showed no significant differences in serum levels of CK , CK-MB , LDH or AST. Histologically, the myocardial tissues of the two models all showed coagulation necrosis and inflammatory cell infiltration, with granulation tissue ingrowth in the infracted tissues and scar formation. The mortality rate of the rats in coronary artery ligation group was 50% at 4 weeks, significantly higher than that of the harmonic scalpel group (20%, P<0.05).

CONCLUSIONCoronary artery ligation and harmonic scalpel method are both effective means to produce rat models of myocardial infarction. The latter method can establish a more stable model for studying the mechanism and treatment of myocardial infarction.

Animals ; Disease Models, Animal ; Male ; Myocardial Infarction ; Rats ; Rats, Sprague-Dawley