Objective To study the association of TCF7L2 gene rs11196218,rs290487 polymorphisms with metabolic syndrome in type 2 diabetes mellitus population.Methods According to the diagnostic criteria of international diabetes federation (IDF),680 cases of type 2 diabetes patients were divided into metabolic syndrome (MS) group and non metabolic syndrome (control) group.DNA was extracted from peripheral mononuclear cells,and then PCR was performed to specifically amplify TCF7L2 gene fragments.Gene polymorphisms were determined by connected enzyme detection reaction.After population representative was checked by Hardy-Weinberg equilibrium,statistical analysis was completed by software SPSS 13.0.Results The population was accorded with Hardy-Weinberg equilibrium and possessed the population representative.Frequency distributions of genotypes (GG,AG and AA) in TCF7L2 gene rs11196218 in MS and control groups were 55.6％(233/419),35.8％(150/419),8.6％ (36/419) and 54.8％ (126/230),39.1％ (90/230),6.1％ (14/230),respectively.Frequency distributions of alleles(G and A) in TCF7L2 gene rs11196218 in MS and control groups were 73.5％(616/838),26.5％(222/838)and 74.3％(342/460),25.7％(118/460),respectively.Frequency distributions of genotypes (GG,AG and AA) in TCF7L2 gene rs290487 in MS and control groups were 14.8％(62/418),42.3％(177/418),42.9％(179/418) and 15.0％(34/226),48.2％(109/226),36.8％(83/226),respectively.Frequency distributions of alleles(G and A) in TCF7L2 gene rs11196218 in MS and control groups were 36.0％ (301/836),64.0％ (535/836) and 39.1％ (177/452),60.9％ (275/452),respectively.Frequency distribution of allele and genotype in TCF7L2 genes rsl 1196218 and rs290487 between the two groups were not associated with metabolic syndrome in type 2 diabetes population (P ＞ 0.05).Conclusions TCF7L2 gene rs11196218,rs290487 polymorphisms has not association with metabolic syndrome of type 2 diabetes.

This study was aimed to investigate the effect of fetal bone marrow-derived mesenchymal stem cells (FBM-MSC) on the development of human Th1 cells. FBM-MSC were isolated, cultured and expanded in vitro. The cells were identified by their phenotype profiles and differential capacity. Human CD4(+) T cells from healthy donors were cultured alone or co-cultured with FBM-MSC (FBM-MSC/CD4). In these two cultures, the quantities of Th1 cells (interferon-γ(+)) were analyzed by flow cytometry. The results indicated that the immunophenotype and multilineage differentiation of FBM-MSC satisfied the generally accepted criteria. FBM-MSC played an inhibitory role in the development of Th1 cells. Flow cytometry analysis showed that the percentage of Th1 cells in FBM-MSC/CD4 was significantly lower than that in CD4(+) T cells cultured alone. The protein level of IFN-γ in FBM-MSC/CD4 detected by ELISA was also lower than that in CD4(+) T cells cultured alone. It was also demonstrated that the expression level of IL-6 in FBM-MSC/CD4 was much higher than that in CD4(+) T cells cultured alone or FBM-MSC. The neutralizing antibody of IL-6 could increase the quantities of Th1 cells and the expression levels of IFN-γ. It is concluded that FBM-MSC may play an inhibitory role in the development of human Th1 cells, and the IL-6 pathway may be one of mechanisms involved in the inhibitory role.
Bone Marrow Cells ; cytology ; Cell Differentiation ; Flow Cytometry ; Humans ; Immunophenotyping ; Interleukin-6 ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Th1 Cells ; cytology

It is commonly accepted that airway hyperresponsiveness (AHR) is a chronic airway inflammation although the exact mechanism of its pathogenesis is still unclear. In the past ten years, an epithelial defect hypothesis has gradually gained supports from the main stream. Airway epithelium is no longer considered only as a simple mechanic barrier but an active interface between the inner and outer environment. Bronchial epithelial cells play a critical role in maintenance of homeostasis in the airway local microenvironment through a wide range of physiologic functions including anti-oxidation, exocrine/endocrine secretions, mucus production and antigen presentation under health and stressed/inflamed/injured conditions. It is reasonably hypothesized that disruption of these functional processes or defects in airway epithelium integrity may be the initial steps leading to airway hyperresponsiveness such as in asthma and chronic obstructive pulmonary disease.
Animals ; Bronchi ; cytology ; Bronchial Hyperreactivity ; physiopathology ; Epithelial Cells ; pathology ; Humans

OBJECTIVETraditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.

METHODSSix virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.

RESULTSThe sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.

CONCLUSIONThe method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.

Escherichia coli Infections ; diagnosis ; microbiology ; Escherichia coli Proteins ; genetics ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Shiga-Toxigenic Escherichia coli ; genetics ; isolation & purification ; Virulence Factors ; genetics

In this study, we constructed the plasmid of Sendai virus (SeV) BB1 strain minigenome with Gaussia luciferase (Gluc) as reporter and compared the rescue efficiency of SeV minigenome mediated by T7 promoter with that by CMV promoter. Firstly, the sequence was designed and synthesized which contained hammerhead ribozyme, sequence composed of the trailer, untranslated region of L gene, untranslated region of N gene, and the leader from SeV, and mutant hepatitis delta virus ribozyme sequence. Then, the synthesized sequence was inserted into pVAX1 containing CMV and T7 promoters and the general vector for SeV minigenome pVAX-miniSeV was obtained. Furthermore, pVAX-miniSeV-Gluc (+) and pVAX-miniSeV-Gluc(-) were obtained by inserting Gluc gene into pVAX-miniSeV. From the supernatant of BHK-21 cell transfected with pVAX-miniSeV-Gluc(+), high level of Gluc expression was detection indicating the normal transcription function of CMV promoter. pVAX-SeV-miniGluc(-) and plasmids expressing N,P and L protein of SeV were co-transfected into BST T7/5 cell which derived from BHK-21 and expressed T7 RNA polymerase stably. And high expression of Gluc was found, which indicated that SeV minigenome was efficiently rescued. However, we failed to repeat the result on BHK-21 cell, implying that T7 promoter and CMV promoter may have different effects on the rescue efficiency of SeV minigenome. Therefore, we further constructed SeV minigenome vectors pT7-miniSeV-Gluc (-) and pCMV-miniSeV-Gluc(-) with single promoter of T7 or CMV. Then, these vectors were transfected into BSR T7/ 5 cells respectively accompanied with the N, P, and L protein expression vectors. The result demonstrated that high expression of Gluc was found in the group of pT7-miniSeV-Gluc(-), which failed in the group of pCMV-miniSeV-Gluc(-). It indicated that T7 promoter significantly increased the rescue efficiency of SeV minigenome. We successfully constructed a SeV minigenome vector with secreted luciferase gene as report er and proved T7 promoter can enhance the rescue efficiency of SeV minigenome, which provides basis for construction of infectious clone containing SeV full-length genome.
Animals ; Cell Line ; Cricetinae ; Cytomegalovirus ; genetics ; DNA-Directed RNA Polymerases ; genetics ; Genome, Viral ; Humans ; Promoter Regions, Genetic ; Respirovirus Infections ; virology ; Sendai virus ; genetics ; physiology ; Viral Proteins ; genetics ; metabolism

OBJECTIVETo evaluate the effect of Chai Shao Liu Jun Tang in combination with Lamivudine for the treatment chronic hepatitis B (CHB) patients.

METHODS405 CHB patients in Guangdong Provincial Hospital were randomly divided into 2 groups, 220 in the treated group, and 185 in the control group. The control group was treated with Lamivudine for 18 months. The treated group was treated with Lamivudine in combination with Chai Shao Liu Jun Tang for 18months. At the 3rd, 6th, 9th, 12th and 18th month during the treatment, the clinical symptoms, ALT normalization rate, HBeAg seroconversion rate, the proportion of patients with undetectable serum HBV DNA, and YMDD mutation rate were observed.

RESULTSALT normalization rates at the 3rd, 6th, 12th, 18th month of the treatment group (69.5%, 85.9%, 90.5%, 82.7%) were higher than those in the control group (50.3%, 65.4%, 78.4%, 69.7%; P < 0.01). HBeAg seroconversion rate, rate of HBV DNA undetectable, and YMDD mutation rate at he 12th and18th month are 77.7%, 57.7%, 25.5%, 6.8%; 86.8%, 74.1%, 33.2%, 8.6% in the treatment group, and 54.6%, 36.8%, 13.0%, 14.6%; 69.2%, 37.3%, 19.5%, 20.5% in the control group (P < 0.01, or P < 0.05).

CONCLUSIONCompared to lamivudine alone, Cai Shao Liu Ju Tang in combination with lamivudine is more effective and induces less YMDD mutation rate in CHB patients.

Adult ; Alanine Transaminase ; blood ; DNA, Viral ; blood ; genetics ; Drug Combinations ; Drug Resistance, Viral ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Genes, Viral ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; administration & dosage ; therapeutic use ; Liver Function Tests ; Male ; Medicine, Chinese Traditional ; Mutation ; Phytotherapy ; Treatment Outcome ; Virus Replication ; drug effects

OBJECTIVETo study separation and purification of flavonids with ethanol/phosphate aqueous two-phase system.

METHODThe diversity of phase separation ability and the distribution of target products in various systems were taken as indicators to analyze aqueous two-phase extraction systems and phase diagrams formed by ethanol and some common salts, screen out EtOH/ K2HPO4 system as the optimla system for extracting total flavonids, and study the impact of proportion of components in EtOH/K2 HPO4 system on the partition coefficient and phase ratio of flavonids.

RESULT AND CONCLUSIONThe EtOH/K2 HPO4 system with omegaEtOH 36.05% and omegaKHPO4 18.20% has been proved as the optimal conditions for separating and purifying total flavonoids of Astragalus (TFA). Under this optimal condition, the partition coefficient and the extraction yield of TFA reached 10.33 and 96.6%, respectively. After extraction, the contents of A. membranaceus saponins and A. membranaceus polysaccharides in top and bottom phases were determined at the same time, showing that A. membranaceus saponins in the removal rate reached 92.01%, and A. membranaceus polysaccharides were totally concentrated in bottom water phase, indicating a removal rate of 100%. Therefore, this is beneficial to separate and purify total flavonids from A. membranaceus crude extracts.

Astragalus membranaceus ; chemistry ; Chemical Fractionation ; methods ; Ethanol ; chemistry ; Flavonoids ; analysis ; isolation & purification ; Phosphates ; chemistry ; Plant Extracts ; analysis ; isolation & purification

BACKGROUNDWith the rapid development of computer technology, digital medicine has become a new direction in surgery. The application of digital medicine in hepatic surgery is still at the early stage and less reported in the literature. The aim of this study was to apply digital medical technology in the context of hepatic surgery.

METHODSData from 64-slice helical computed tomography of 17 patients, including 13 with hepatocellular carcinoma and 4 with hepatic hemangioma, were imported into independently developed medical image software program, segmentation and three-dimensional reconstruction were performed. The three-dimensional models were then processed with the FreeForm Modeling System. We used virtual surgical instruments to perform surgery on the models. Simulated surgeries included six hepatic segmentectomies, four left hemihepatectomies, three right hemihepatectomies for hepatocellular carcinoma, one hepatic segmentectomy, two stripping surgeries, and one irregular segmentectomy combined with stripping surgery for hemangioma. For resections involving more than three hepatic segments, total and residual functional hepatic volumes were measured before and after simulation surgery, and the resection ratio was calculated.

RESULTSThe anatomy of the models was distinct and was used to localize lesions. We used virtual surgical instruments to perform simulated surgeries and used the models to optimize actual surgeries. We were able to minimize resection volume as well as surgical risk.

CONCLUSIONSDigital medical technology is helpful in the diagnosis of hepatic disease and in optimizing surgical plans. Three-dimensional models can decrease surgical risk and help prevent postoperative hepatic failure.

Adult ; Aged ; Female ; Humans ; Liver ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods

Objective To provide scientific evidences for improving children early integrated development,through analyzing the mental developmental status and characteristics of 322 cases of 2-year-old children in Nanjing.Methods Intelligence and motor development condition in 322 cases of 2-year-old children were assessed by using Bayley Scales of Infant Development test,and the assessed results were analyzed.Results 1.The incidences of the children whose mental development index(MDI)or psychomotor development index(PDI)were under 69 were 3.1% and 5.6%,respectively;2.The MDI mean score(114.34?19.65)was significantly higher than that of PDI(101.73?21.53)(t=9.71,P0.05).Conclusions The incidences of mental retardation in this study were consistent with the result reported by World Health Organization.There were differences between motor and intelligence development in children,as well as the intelligence development between male and female.Therefore,it should be implemented early childhood developmental screening in child health care.Parents should be given scientific guides about intelligence and motor development of children.

OBJECTIVETo study the correlations between clinical features and liver pathohistological changes of chronic hepatitis B virus (HBV) carriers and to discuss the factors which may influence the prognosis.

METHODSNinety HBV carriers who had liver biopsies were enrolled in this study.

RESULTS(1) The mean follow-up period of the patients was 118 weeks. (2) Fifty-four patients (60.0%) had G1 hepatitis and 21 (23.3%) had G2 hepatitis. The fibrosis stages were graded as S1(42) and S2(21). (3) There were significant age differences among S0, S1 and S2. (4) There were significant differences in aminotransferase levels between patients who had a normal liver histology and those who had mild hepatitis. (5) The grades of liver inflammation were not correlated with the titers of HBeAg and HBV DNA in sera. The stages of liver fibrosis were not correlated with the titers of HBVDNA in sera. Most of the HBeAg negative patients progressed to S2. (6) There were significant differences in spleen dimensions measured by ultrasonography between S0, S1 and S2 patients. (7) During the follow-up period serum aminotransferase (ALT) levels remained normal in 60 patients (group A); 22 patients had transient elevations (group B), and 8 patients had persistent increases (group C). There were significant differences of the ratios of S0 and S2 cases among patients in groups A, B and C. (8) Age and fibrosis stages were predictive factors of liver cirrhosis.

CONCLUSIONSMost chronic HBV carriers had mild inflammatory histological changes in their livers and also had different degrees of liver fibrosis. This follow-up study shows that some of those carriers should have had antiviral therapy.

Adult ; Carrier State ; diagnosis ; pathology ; virology ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; diagnosis ; pathology ; Humans ; Liver Cirrhosis ; diagnosis ; pathology ; virology ; Male ; Middle Aged ; Prognosis