Objective To investigate the insulin and glucagon levels in the patients with different islet cell antibody (ICA) subtypes and to explore the pathogenesis of latent autoimmune diabetes of adult (LADA). Methods Subjects were classified by immunohistology and 29 ICA-peripheral-positive DM patients, 28 ICA-diffused-positive DM patients and 17 controls (ICA-negative) were included. Serum glucose, insulin and plasma glucagon were measured at 0, 30, 60, 120 min after standard meal. Results (1) As compared with controls, glucagon in ICA positive groups were higher (both P

Objective To investigate the distribution status of cytochrome CYP2C19 polymorphisms among patients with cardio-vascular diseases in Dongguan area .Methods A total of 1662 patients with cardiovascular diseases (CVD) in our hospital were se-lected .The peripheral blood was collected for extracting genomic DNA .The CYP2C19 genotype was detected by the polymerase chain reaction (PCR) combined with gene chip technology .The allele frequency and metabolic phenotype of CYP2C19 were com-pared between the CVD patients aged lower than 45 years and aged higher than 45 years old .Results Among 1662 cases ,713 cases (42 .90% ) were CYP2C19 extensive metabolizer ,740 cases(44 .52% ) were moderate metabolizer and 209 cases (12 .58% ) were poor metabolizer .The allele frequencies of CYP2C19﹡1 ,CYP2C19﹡2 and CYP2C19﹡3 were 65 .16% ,30 .08% and 4 .75% re-spectively .One hundred and four cases (40 .00% ) of extensive metabolisms were detected in the lower than 45 years old group ,104 cases(45 .38% ) of moderate metabolisms and 38 cases (14 .62% ) were detected .In the higher than 45 years old group ,609 cases (43 .44% ) of extensive metabolisms ,622 cases of moderate metabolisms and 171 cases(12 .20% ) of poor metabolism were detec-ted .The proportion of various genotypes had no statistical difference between the lower than 45 year old group and higher than 45 year old group(P>0 .05) .Conclusion Detecting CYP2C19 genotype for determining the genetic characteristics can evaluate the clopidogrel resistance risk and formulate the personalized anti-platelet therapeutic scheme .

AIM: To deplore the adjustion of whole peptidoglycan of bifidobacterium bifidum to functions of macrophages.METHODS:Levels of interleukin 6, interleukin 12 produced by peritoneal macrophages of nude mice was detected by ELISA method, and the content of nitric oxide (NO) was detected by using Griess reagent, after whole peptidoglycan were injected into nude mice peritoneally. RESULTS:The content of interleukin 6, interleukin 12 and NO secreted from peritoneal macrophages of nude mice in the whole peptidoglycan injection group was 732.54?190.30(pg/mL)、816.37?96.40(pg/mL) and 48.90?6.51(?mol/L) respectively.It was 303.78?171.75(pg/mL), 510.27?123.46(pg/mL) and 30.67?12.83(?mol/L) respectively in the control group. In view of the content of interleukin 6, interleukin 12 and NO, there existed significant difference when whole peptidoglycan injection group was compared with control group( P

Objective To study the feasibility,effectiveness and safety of temporary pacing at the right ventricular outflow tract area in patients with inferior acute myocardial infarction(AMI)and two or three degree atrioventricular block(AVB). Methods Ninety-five patients with inferior AMI and two or three degree AVB admitted into Cangzhou People's Hospital were randomly divided into right ventricular apex pacing group(47 cases) and right ventricular outflow tract region pacing group(48 cases). Seldinger method was used to perform the right femoral vein puncture in which a 6F sheath tube was placed,and a diode temporary pacing electrode was introduced into it in the two groups. Under the X-ray guidance,in the right ventricular apex pacing group,the electrode was pushed from the inferior vena cava to enter into the right atrium,then cross the tricuspid and finally reach the right ventricular apical portion pacing. In the right ventricular outflow tract region pacing group,under the X-ray guidance, after the electrode was withdrawn and rotated slightly clockwise,it was sent forward to the right ventricular outflow tract region pacing. The duration from the puncture at the right femoral vein to the implanted electrode reaching the pacing region,the pacing threshold,the un-favorable pacing,the incidence of ventricular arrhythmia and prognosis were observed in the two groups. Results The pacing time and pacing threshold in right ventricular apex pacing group were obviously higher than those in the right ventricular outflow tract region pacing group〔pacing time(s):336±150 vs. 354±152,pacing threshold(V):0.9±0.4 vs. 0.7±0.3,both P<0.05〕. The mortality in intensive care unit (ICU)〔0(0/48)vs. 2.13%(1/47)〕,the incidence of bad pacing〔10.42%(5/48)vs. 17.02%(8/47)〕and ventricular fibrillation(VF),sustained ventricular tachycardia,contraction of non-sustained ventricular tachycardia or frequent ventricular premature beat(VPB)and other arrhythmia incidence of complications〔10.42%(5/48)vs. 12.77%(6/47)〕in the right ventricular outflow tract region pacing group were all lower than those in the right ventricular apex pacing group. Conclusion The application of right ventricular outflow tract pacing in patients with inferior AMI and two or three degree AVB is safe and effective,and its stability is fine.

Objective To investigate the drug resistance tendency of Klebsiella pneumoniae isolated from the clinical samples during 2012-2014 to provide reliable evidence for clinical treatment .Methods The Klebsiella pneumoniae strains isolated from the submitted specimens were collected and identified according to the national clinical test procedures ,and the drug sensitivity test was performed by using MIC method .The confirmation test of ESBLs was conducted by using K‐B method and the phenotype of carbap‐enemases producing was confirmed by using the improved Hodge test .Results Totally 410 strains of Klebsiella pneumoniae were i‐solated ,55 .87% of which were derived from sputum ,and the rest was derived from pus(9 .53% ) ,secretion(9 .47% ) and blood (8 .78% );Klebsiella pneumoniae was mainly originated from ICU ,respiration department and oncology department ,accounting for 16 .10% ,9 .02% and 7 .80% respectively ;the resistance rates of Klebsiella pneumoniae against imipenem was 0 .74% ,the resist‐ance rates of Klebsiella pneumoniae against ampicillin/sulbactam ,cafazolin ,cefepime ,cefotaxime ,cefatriaxone ,ceftazidine ,compound sulfamethoxazole were decreased year by year ,while which against amoxicillin/clavulanic acid showed the increasing trend as a whole .Conclusion Timely conducting the identification and drug susceptibility analysis on local Klebsiella pneumoniae and tracking its drug resistance trend can guide the rational and standardized use of antibacterial drugs ,reduces the pressure for selecting anti‐bacterial drugs in order to reduce the generation of drug resistant strains .

Objective To identify the hemolysin BL(HBL) gene from Bacillus cereus of patients with endophthalmitis comfirmed by API bacterial identification test strip,and detect its biological activity in vitro.Methods Three pairs of specific primers were designed according to the gene sequence of HBL(B,L1 and L2 components),then the PCR assay were established through condition optimization,and to further detect five Bacillus cereus strains isolated from clinical patients with endophthalmitis.HBL with biological activity was extracted by salt fractionation from a randomly selected strain,and a series of different concentrations of HBL were prepared and acted on sheep red blood cells(SRBC),Vero cells and Hela cells; virulence of HBL was also assessed through observating lethal effect of which on mice with intraperitoneal injection.Results Three genes of HBL were detected in all B.cereus strains from clinical patients; Strong hemolytic activity of HBL showed obvious time-and dose-dependent.In the study,we found the morphological changes of Vero and Hela cells caused by HBL were different,but cell death were the same result with contents released; Within 48 h after infection,lethality of HBL for mice showed 100％ with the concentration of more than 2.0 HU/ml,and was also in a time-and dose-dependent manner.Conclusion HBL isolated from B.Cereus had high hemolysis activity and low concentration.The expression of all BL genes provided a strong basis for the clinical feature of B.Cereus infection,which was developed rapidly and with a poor prognosis.It also provided a new method for rapid diagnosis and molecular epidemiology investigation in clinical.

BACKGROUND: Tumor necrosis factor-α (TNF-α) is one of important cytokines to promote the maturation of dendritic cells. Blockage of TNF-α action by binding with soluble tumor necrosis factor receptor 1 (sTNFR1) may arrest dendritic cells in an immature state and induce stable, long-term tolerance. OBJECTIVE: To construct the lentiviral vectors carrying sTNFR1 gene and investigate sTNFR1 expression in immature dendritic cells. METHODS: Total RNA of human peripheral blood mononuclear cells was taken as a template. The sTNFR1 gene fragment was amplified by RT-PCR, subcloned to the lentiviral vectors pXZ208, and ligated to the enhanced green fluorescent protein (eGFP) reporter gene to establish lentiviral vector, called pXZ9-sTNFR1. DNA sequencing was performed for lentiviral vector identification. Lentivirus was prepared by transfection of 293 FT cells with pXZ9-sTNFR1. Viral titer was determined by eGFP expression. C57BL/6 mouse bone marrow-derived dendritic cells were in vitro cultured with low-dose granulocyte-macrophage colony stimulating factors and interleukin 4. On day 5 of culture, immature dendritic cells were transfected with pXZ9-sTNFR1 recombinant lentiviral supernatant, sTNFR1 transcription was detected by RT-PCR, sTNFR1 protein expression by Western blot analysis. Following sTNFR1 gene modification and lipopolysaccharide stimulation, the phenotype characteristics of dendritic cells were observed. RESULTS AND CONCLUSION: Recombinant plasmid pXZ9-sTNFR1 was successfully constructed. Twenty-four hours after 293 FT cell transfection, eGFP expression was observed and viral titer was over 10<'6> U/L. RT-PCR demonstrated that pXZ9-sTNFRl-transfected immature dendritic cells showed sTNFR1 positive expression. Western blot analysis revealed that sTNFR1 protein appeared in the immature dendritic cells and supernatant following 293 FT cell transfection. On day 5 of culture, dendritic cells expressed low level of class Ⅱ major histocompatibility complex (MHC Ⅱ), as well as CD40, CD86, CD80, molecules. However, following lipopolysaccharide stimulation, dendritic cells expressed high level of MHC Ⅱ, as well as CD40, CD80, and CD86, molecules, exhibiting the phenotype characteristics of mature dendritic cells. But after sTNFR modification, the expression level of MHC Ⅱ, as well as CD40, CD80, and CD86, molecules was not altered obviously. Lentiviral vectors carrying sTNFR1 gene and eGFP reporter gene were successfully constructed, and recombinant lentiviral plasmids with high titer were acquired. Following high efficacy of lentiviral gene transfection, immature dendritic cells stably express sTNFR1 mRNA and protein, which prevents immature dendritic cells from activation by exogenous lipopolysaccharide and maintains the immature state.

Objective To determine the relationship of cytochrome P-450 epoxygenase metabolites of arachidonic acids with impaired endothelium-dependent vasorelaxation in the healthy elderly. Methods A case-control study was employed.The study enrolled 60 healthy young adults (group A) and 60 healthy senior citizens (group B) of Han population in Shanghai. Serum contents of 14,15-dihydroxyeicosatrienoic acid (14,15-DHET) (a stable metabolite of 14,15-EET), 6-Kote-PGF1a (a stable metabolite of prostaglandin) and TXB2 (a stable metabolite of thromboxane A2) were measured using ELISA kits. The endothelium-dependent vasorelaxation (EDV) and endothelium-independent vasorelaxation (NEDV) in brachial arteries were determined by color Doppler ultrasound. Results Compared with group A, group B had significant higher levels of hemoglobin A1c, triglyceride and cholesterol levels (P＜0.01), and significant lower levels of 14,15-DHET and 6-Kote-PGF1a (P＜0.001), leading to increased values of TXB2/6-Kote-PGF1a and TXB2/14,15-DHET. There was bigger basal interior diameter of brachial arterials with reduced EDV and NEDV response (P＜0.01 and P＜0.05, vs. group A respectively) in group B. Moreover, the age was negatively correlated with 14,15-DHET and TXB2/14,15-DHET. Conclusions Our results indicate that the impaired EDV and NEDV in aging are associated with reduced production of arachidonic acid metabolites through cytochrome P-450 epoxygenase pathway and cyooxygenase pathway of endothelium in the healthy elderly.

Objective:To explore the association of the genes GSTM1 and GSTT1 with oligozoospermia infertility.Methods:PCR technique was used to analyze the GSTM1 and GSTT1 genes polymorphisms in 75 with oligospermia infertility and 36 healthy individuals of Zhuang population from Guangxi Baise area,and then the possibility function of GSTM1 and GSTT1 genes in human oligozoospermia were studied.Results:Analyses of the polymorphisms in GSTM1 and GSTT1 genes showed that GSTM1 defect or combined defects of both GSTM1 and GSTT1 was found more frequent in patients with infertile oligospermia than in the healthy control.Conclusion:GSTM1 and GSTT1 genes may have modulating effects on human spermatogenesis,whose mechanism needs further studies.

Objective:To develop a two-step method for purification of monoclonal antibody rhTF243 protein from mouse ascites by using hydrophobic charge induction chromatography(HCIC) and affinity chromatography with protein A sepharose CL-4B.Methods:The ascites was first purified by HCIC after centrifugation and filtration.Then the fraction containing the protein of interest was directly purified by affinity chromatography with protein A sepharose CL-4B.Results:The purity of the obtained monoclonal antibody was up to 97% with recovery of 73% and of high activity.Conclusion:The method for purification of monoclonal antibody is developed using HCIC and Protein A affinity chromatography and the obtained antibodies are of high purity and activity.