Objective To investigate the relationship between adiponectin and the first-phase of pancreatic P-cell insulin secretion in subjects with different statuses of glucose tolerance. Methods Thirty-seven patients with newly diagnosed type 2 diabetes mellitus (DM) , 30 patients with abnormal glucose tolerance (IGR) , and 40 normal control subjects (NGT) underwent intravenous glucose tolerance test (IVGTT). Fasting adiponectin and proinsulin (PI) was assayed by EL1SA. Fasting free fatty acid ( FFA) was measured by colorimetry. Insulin area under the curve ( AUC ) , incremental AUC (iAUC) from 0 min to 10 min, AIR3-5, homeostasis model assessment for insnlin resistance (HOMA-IR) , and for β cell function ( HOMA-p) were calculated. The relationship between adiponectin and AUC, iAUC, AIR3-5, proinsulin, FFA, and HOMA-IR was explored. Results (1) The levels of AUC, iAUC, AIR3-5, and adiponectin in DM group and IGR group were significantly lower than those in NGT group (P<0.05), reduced in DM group than those in IGR group(P<0.05). (2) The levels of PI in DM group and IGR group were significantly higher than that in NGT (P<0.05). (3) Adiponectin was positively correlated with HOMA-p,AUC,iAUC,AIR3-5, and HDL-C,while negatively correlated with proinsulin, HOMA-IR, and LDL-C. (4) Proinsulin was positively correlated with HOMA-IR. (5 ) Multiple regression stepwise analysis showed that adiponectin was independently associated with AUC. Conclusions Adiponectin was an independent factor affecting the first phase of pancreatic p-cell insulin secretion. Low adiponectin level could predict the dysfunction of the first phase pancreatic p-cell secretion as well as insulin resistance in patients with type 2 diabetes.

CD8-Positive T-Lymphocytes ; metabolism ; Case-Control Studies ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; biosynthesis ; Lymphocytes ; immunology ; Oligodeoxyribonucleotides ; pharmacology

AIM: To establish an animal model of experimental autoimmune myocarditis (EAM) in BALB/c mice and to investigate the expression and significance of Toll-like receptor 3 in mouse EAM. METHODS: BALB/c mice were immunized with cardiac myosin extracted from porcine ventricular myocardium covered by complete freund's adjuvant (CFA) on 0 d and 7 d, then divided into immunized with CFA only. Serum and myocardium samples were collected at 14 d and 21 d after the first immunization. HE staining was used to identify the areas of inflammation. The myosin IgG antibody was examined by indirect ELISA assay. The changes of TLR3 protein and mRNA expression in myocardial tissue were measured by immunohistochemistry and real time-PCR. RESULTS: Compared to control group, immunohistochemistry results showed that there was positive expression of TLR3 in the myocardium of mice with EAM and the mRNA of TLR3 were more than 20 times (P<0.05). The expression of interferon beta mRNA in EAM group was more than 14 times as many as basal expression, that of tumor necrosis factor alpha was more than 18 times (P<0.05). CONCLUSION: The expression of Toll-like receptor 3 in myocardium is up-regulated in experimental autoimmune myocarditis. The inflammatory response to cardiac myosin may associate with the TLR3 signal transduction pathway.

OBJECTIVETo study the molecular types of Staphylococcus aureus isolated from a severe food-poisoning and to trace the possible strains.

METHODSReal-time PCR was applied to detect nuc gene as a specific marker for S. aureus, mecA gene encoding methicillin resistance and 5 other genes encoding staphylococcal enterotoxins (sea, seb, see, sed, see). Isolates were also performed with 16S rRNA oligonucleotide sequence analyzing by DNAStar MegAlign 5.0 software and pulse-field gel electrophoresis (PFGE) by BioNumerics Version 4.0 software.

RESULTSThe nuc gene was detected from the 10 isolated strains, sea and seb genes were detected from 7 strains. There were 4 16 S rRNA types and 5 PFGE types found from all the strains.

CONCLUSIONSThree relative S. aureus strains were involved in the severe food-poisoning at least. Molecular subtyping might give a molecular epidemiological evidence and support the source tracing of an outbreak.

Bacterial Typing Techniques ; China ; Electrophoresis, Gel, Pulsed-Field ; Enterotoxins ; Humans ; Staphylococcal Food Poisoning ; epidemiology ; microbiology ; Staphylococcus aureus ; classification ; genetics ; isolation & purification

Objective To investigate the risk factors of clopidogrel resistance in type 2 diabetes mellitus and cerebral infarction patients.Methods A total of 112 patients were divided into treatment group (clopidogrel resistant group,n =25) and control group (clopidogrel non-resistant group,n =87) according to the platelet aggregation.All the patients were taken clopidogrel 75 mg daily at least 7 d.The general data of the patients were recorded.The National institute of health stroke scale (NIHSS) and Alberta stroke program early computed tomography (ASPECT) score were recorded in two groups.The platelet aggregation function were analyzed by adenosine diphosphate (ADP) induced optical turbid metric method and analyze their correlation.Results The NIHSS in treatment group and control group were (3.91 ± 4.95),(2.40 ± 2.09) point,the ASPECT score in treatment group and control group were (11.57 ±6.05),(13.40 ± 1.29) point,with significant difference (P ＜ 0.05).Conclusion Clopidogrel resistance is associated with increased clinical severity and infarct volume in type 2 diabetes mellitus and cerebral infarction patients.

Objective To know the information about the past disease history and HBV infection in the male hit-the-pipe patients. Methods 452 male addicts that are accepting the treatment of abandoning drug habits in a drug-relief reformatory in Changsha City were randomly taken as the subjects, A questionnaire about the past disease history and daily life contact history was dispatched and collected. ELISA assay was used to detect the mark of the HBV infection and the liver function of the subjects. Results There was a statistical relationship between whether the persons in the studied sample have a past disease history of other viral hepatitis, and the HBV total infection, HbsAg infection, and large triple yang and little triple yang infections. Whether the addicts accepted blood products is statistically related to the HbsAg infection rate. Whether the addicts pricked holes on their ears for earrings is related to HbsAg infection, and little triple yang infection. Conclusions It is the most important influencing factors for HBV infection that the male addicts have a disease history of other viral hepatitis and other related medical history.

OBJECTIVETo investigate the effects of microwave irradiation on the expression and regulation of heat shock proteins (HSPs) in primary cultured rat hippocampal neurons.

METHODSNeurons were exposed to 90 mW/cm(2) microwave irradiation for 10 minutes. Western blot was used to determine the expression of HSP27, HSP70, HSP90 and heat shock factor 1 (HSF1) at 0, 3, 6, 12 and 24 hour respectively. Real-time RT-PCR was used to determine the mRNA expression of HSF1. DNA-binding activity of HSF1 was measured by electrophoretic mobility shift assay (EMSA).

RESULTSThe protein expression of HSP27 was significantly increased by 22%, 36%, 18% at 3, 6, 12 h, respectively (P < 0.05). The protein expression of HSP70 was significantly increased by 23%, 32%, 26% at 3, 6, 12 h, respectively (P < 0.05, P < 0.01). The protein expression of HSP90 was significantly increased by 27%, 33% at 6, 12 h, respectively (P < 0.05, P < 0.01). The DNA-binding activity of HSF1 was stimulated, however, no significant change of the expression of HSF1 was observed on both the mRNA and protein levels.

CONCLUSIONThe transcriptional activity of HSF1 is activated by microwave irradiation, which promotes the expression of HSPs. Heat shock response which contributes to establish a cytoprotective state is induced by microwave irradiation in primary cultured rat hippocampal neurons.

Animals ; Cells, Cultured ; Heat-Shock Proteins ; metabolism ; Hippocampus ; metabolism ; radiation effects ; Microwaves ; adverse effects ; Neurons ; metabolism ; Rats

OBJECTIVETo study the effects of Qingli Shengjing Pills (QSP) on the apoptosis of germ cells and expressions of Fas and FasL in male mice infected with Escherichia coli (E. coli), and to clarify the molecular mechanism of QSP in the treatment of male infertility induced by E. coli infection.

METHODSFifty male mice were injected with E. coli via the bladder to make infection models, and at 15 dpi equally randomized into five groups: an untreated, a high-dose QSP (22.5 g/ml), a medium-dose QSP (13.5 g/ml), a low-dose QSP (4.50 g/ml) and a Furadantin treatment group, which were coded as MN, MTa, MTb, MTc and MTd, respectively. Another 10 mice were injected with saline and included in the control group coded as CT. After 10 days of oral medication, the apoptosis of germ cells in the testis of the mice was detected by flow cytometry, the expressions of Fas and FasL determined by immunohistochemistry and the histopathological changes observed simultaneously.

RESULTSAfter the treatment, the apoptosis of germ cells was observed in all the infected groups, and the apoptosis level in MN (57.44%) was significantly higher than that in CT (28.54%), MTb (28.59%) or MTa (30.11%) (P < 0.01) but had no significant difference from that in MTc (46.54%) or MTd (43.41%) (P > 0.05). The expressions of Fas and FasL proteins were significantly higher in MN than in CT, MTa, MTb, MTc and MTd (P < 0.01). Histopathological changes of the testis tissue were observed in MN, but not in other groups.

CONCLUSIONE. coli infection could increase the apoptosis rate of germ cells and the expressions of Fas and FasL proteins. Qingli Shengjing Pills, capable of enhancing reproductivity by reducing the expressions of Fas and FasL and apoptosis of germ cells, can be used as one of the effective drugs for infertility induced by E. coli infection.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Escherichia coli ; Fas Ligand Protein ; metabolism ; Infertility, Male ; drug therapy ; metabolism ; microbiology ; Male ; Mice ; Mice, Inbred Strains ; Testis ; cytology ; drug effects ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo apply multiplex polymerase chain reaction (MPCR) assay and sequencing in study of the carrying status of four pathogenicity-related genes of Vibrio cholerae (V.cholerae) and the variation of ctxA.

METHODSPrimers targeting cholera toxin sub-unit A gene (ctxA), toxin-coregulated pilus gene (tcpA), accessory cholera enterotoxin gene (ace), zonula occludens toxin gene (zot) were designed and the MPCR method was applied to detect the pathogenicity-related genes of 276 strains of V.cholerae isolates. The amplified fragments of ctxA gene were sequenced and the genetic homology of the amplified fragments of ctxA was analyzed.

RESULTSOf the 276 strains of V.cholerae, 93.9% strains from human sources belong to the pathogenicity-related genes type A (ctxA(+)tcpA(+)ace(+)zot(+) type) and 6.1% belong to pathogenicity-related genes type C (ctxA(-)tcpA(-)ace(-)zot(-) type). Type A strains from clinical sources were isolated from patients with mild to severe symptom and carriers, among which 68.5% were isolated from patients with mild symptom and 21.9% from carriers. All 63.6% of type C strains from clinical sources were isolated from patients with mild symptom and 36.4% from carriers. The proportion of type C strains that caused mild symptom was higher than that of type A strains. Of the 78 strains isolated from the environment, 9.0% strains belong to pathogenicity-related type A and 35.9% belong to the pathogenicity-related genes type B (ctxA(-)tcpA(-)ace(+)zot(+) type), while 55.1% belong to pathogenicity-related genes type C. The sequencing results showed little genetic variation among the amplified fragments for ctxA.

CONCLUSIONMPCR disclosed the polymorphic status of pathogenicity-related gene patterns in V.cholerae isolates of Guangzhou, providing effective means for further study on evolution of pathogenicity-related genes among V.cholerae isolates from human and environmental sources. This study also offers significant guidance for effective prevention, control and warning against cholera epidemic in local area.

China ; Cholera Toxin ; genetics ; DNA, Bacterial ; Genes, Bacterial ; genetics ; Genotype ; Humans ; Polymerase Chain Reaction ; Sequence Analysis ; Vibrio cholerae ; classification ; genetics ; isolation & purification

OBJECTIVETo construct different mutants of human p53 for expression in eukaryotic cells and investigate the effects of these mutants on stress-induced cell apoptosis.

METHODSHuman p53 cDNA was amplified by PCR and cloned into pcDNA3/HA vector following the routine procedures. The Ser15 and Ser46 of p53 were mutated to Ala and identified by enzyme digestion and PCR, and these mutants were expressed in NIH3T3 cells and detected by Western blotting. After transfection with the plasmids of different p53 mutants, the NIH3T3 cells were double-stained with AnnexinV-FITC and propidium iodide for apoptotic analysis using flow cytometry.

RESULTSThe recombinant plasmids of HA-tagged wild-type p53, HA-p53(WT), and its mutants, HA-p53(S15A) and HA-p53(S46A), were successfully constructed and expressed efficiently in NIH3T3 cells. The apoptotic ratio of p53(WT)-transfected cells induced by arsenite increased and that of p53(S15A)-transfected cells decreased significantly after arsenite stimulation, but no significant changes occurred in the apoptosis of p53(S46A)-transfected cells.

CONCLUSIONThe phosphorylation on Ser15 of p53 plays an important role in mediating arsenite-induced cell apoptosis.

Animals ; Apoptosis ; drug effects ; Arsenites ; pharmacology ; Base Sequence ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Humans ; Mice ; Molecular Sequence Data ; Mutation ; NIH 3T3 Cells ; Phosphorylation ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism