Objective To investigate the effects of tumor necrosis factor-alpha (TNF-?) on insulin sensitivity and glucose-lipid metabolism in TNF-?-induced IR mice. Methods Male C57BL/6J mice were given an intraperitoneal injection of TNF-? (H group,6?g/kg; M group,3?g/kg; L group,1?g/kg;twice daily) and saline (NC group) for 7 days. The plasma glucose and insulin were assayed during intravenous glucose tolerance test (IVGTT) and hyperinsulinemic-euglycemic clamp combined with 3-[3H] glucose as a tracer was carried out. Results After TNF-? treatment,fasting blood glucose (FBG),plasma insulin and free fatty acids (FFA) were significantly elevated in H group compared with NC,L and M groups (P

Objective To investigate the roles of myeloid differentiation factor 88 (MyD88) and TIR domain-containing adaptor inducing interferon-β (TRIF) in sepsis-induced myocardial dysfunction, and to analyze whether strain rate (SR) can be early sensitive evaluation for septic heart failure.Methods Sixty-four healthy male C57BL/6 mice were divided into four groups by random number table (n = 16 in each group): sham group, cecum ligation and puncture (CLP)-induced sepsis model group, anti-MyD88 group and anti-TRIF group. The anti-MyD88 group and anti-TRIF group were injected with 5μL/g of anti-MyD88 antibody or anti-TRIF antibody through the tail veins 2 hours before CLP. Eight animals in each group were used to observe the survival of 24 hours, and the other 8 myocardial tissues were harvested for examination. The cardiac function was evaluated by echocardiography before and 6 hours and 12 hours after operation. The mRNA expressions of MyD88, TRIF and inflammatory factors in myocardium were measured by polymerase chain reaction (PCR) at 24 hours after operation, and the degree of neutrophils infiltration was detected by myeloperoxidase (MPO) activity.Results The number of 24-hour survive in anti-MyD88 group and anti-TRIF group were higher than that in CLP group (number: 4, 3 vs. 2,P = 0.044,P = 0.047). Compared with sham group, the cardiac function was significantly decreased, the mRNA expressions of myocardial tissues MyD88, TRIF, interleukin (IL-1, IL-6) and tumor necrosis factor-α (TNF-α) were significantly increased, and the infiltration of neutrophils were obvious in CLP group. Compared with CLP group, the left ventricular short axis fractional shortening rate (FS) and SR were significantly increased after 12 hours in anti-MyD88 group and anti-TRIF group [FS: (49.52±1.78)%, (49.89±1.49)%vs. (41.11±1.63)%, SR (s-1): 17.63±2.16, 17.85±1.64 vs. 12.55±1.84]; the mRNA expressions of MyD88, TRIF and inflammatory factors were significantly decreased [MyD88 mRNA (A value): 0.463±0.046, 0.505±0.048 vs. 0.638±0.102, TRIF mRNA (A value): 0.413±0.031, 0.410±0.021 vs. 0.625±0.057, IL-1 mRNA (A value):0.569±0.101, 0.570±0.091 vs. 0.946±0.171, IL-6 mRNA (A value): 0.551±0.143, 0.431±0.157 vs. 0.850±0.194, TNF-α mRNA (A value): 0.471±0.082, 0.444±0.093 vs. 0.707±0.094]; and the infiltration of neutrophils were significantly decreased [MPO (U/L): 62.34±2.60, 60.87±2.40 vs. 73.83±4.90], with statistically significant differences (allP < 0.05). There was no statistical difference in above parameters between the anti-MyD88 group and anti-TRIF group (allP > 0.05).Conclusions Blocking MyD88 and TRIF expression play significant and similar roles in protecting cardiac deterioration from sepsis by attenuating cytokine release, reducing neutrophil infiltration. SR can sensitively assess septic cardiac dysfunction.

Polydatin is a monocrystaline compound isolated from Polygonum cuspidatum Sieb. et Zucc. (Polygonaceae) with biological properties, such as anti-inflammation, anti-oxidative and nephroprotective effects. Increasing number of studies have demonstrated the protective effect of polydatin on renal ischemia reperfusion injury. However, the possible mechanisms of this protection are not fully elucidated. This study aimed to investigate the effect of polydatin on ischemia-reperfusion induced expression of toll-like receptor4 (TLR4) in rat renal tubular epithelia cells (NRK-52E), and analyze the mechanism of polydatin on TLR4 signal pathway. The cultured NRK-52E cells were incubated in three gas incubators for a period of 6 h at hypoxia and 24h at reoxygenation to simulate the ischemia-reperfusion injury in vitro. TLR4 mRNA level was analyzed by real-time-PCR, and the protein expression of TLR4 and NF-κB by Western blotting, while TNF-α and IL-1β proteins expressions were detected by ELISA. Polydatin downregulated I/R induced mRNA and protein expressions of TLR4, and decreased the protein expression of NF-κB, TNF-α and IL-1β. The TLR4 blocker partially antagonized the effect of I/R on NF-κB signaling, and such inhibitory effect was markedly enhanced by polydatin. In the present study, polydatin protects NRK-52E cells from I/R injury possibly by relieving the inflammatory response through regulation of TLR4/NF-κB signaling pathway.
Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; drug effects ; Glucosides ; pharmacology ; Humans ; Rats ; Reperfusion Injury ; drug therapy ; genetics ; metabolism ; Stilbenes ; pharmacology ; Toll-Like Receptor 4 ; genetics ; metabolism

Objective To investigate the clinical characteristics, the association with the risk factors of macrovascular complications, the prevalence of microvascular complications between LADA patients and Type 2 diabetes(T2DM). Methods A cross-sectional study of 30 cases of LADA(ICA positive) patients and 60 cases of ICA negative T2DM subjects matched with age, sex and duration.Results As compared with T2DM, LADA cases showed a lower BMI(21 3?0 5 vs 23 3?0.4kg/m 2, P

Objective To investigate the expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse (POP). Methods Between November 2006 and June 2008, 68 patients with POP underwent surgical treatment in Shengjing Hospital of China Medical University were enrolled in this study, who were classified into 10 patients with grade Ⅰ , 21 patients with grade Ⅱ , 25 patients with grade Ⅲ and 12 patients with grade Ⅳ in accordance with pelvic organ prolapse quantitation (POP-Q). Meanwhile, 18 cases with early cervical cancer at stage of Ⅰ b were treated by total hysterectomy and bilateral salpingo-oophorectomy, their anterior vaginal tissues were selected as controls. Immunohistochemical staining was performed to detect the expression of elastin and fibulin-5. Results (1 ) Elastin and fibulin-5 were mainly expressed at extracellular matrix(ECM). (2)The positive rate of fibulin-5 expression in anterior vaginal wall were 5% (2/37 ) in grade Ⅲ/Ⅳ and 26% (8/31 ) in grade Ⅰ/Ⅱ POP patients, which reached statistical difference (P = 0. 035). However, no statistical different expression was found between postmenopausal (13%, 8/60) and non-menopausal patients (2/8), vaginal delivery ≤ 2 (19%, 5/27 ) and ＞ 2 patients (12%, 5/41, P ＞ 0. 05 ). (3 ) The positive rate of elastin expression in anterior vaginal wall in POP group was 31% (21/68), which was significantly lower than 72% (13/18) of control group (P = 0. 002). Among POP group, 19% (7/37) of elastin expression in grade Ⅲ/Ⅳ POP was significantly lower than 45% (14/31 ) in grade Ⅰ/Ⅱ of POP patients. However, no statistical difference was found between postmenopausal (30%, 18/60 ) and non-menopausal patients (3/8 ), vaginal delivery ≤ 2 (26%, 7/27) and ＞2 patients(34%, 14/41, P＞0. 05). (4)In POP group, both positive expression of fibulin-5 and elastin of anterior vaginal wall was in 6 cases, both negative expression of fibulin-5 and elastin was in 43 cases. It was illustrated that elastin and fibulin-5 had an positive relationship (P = 0. 031 ). Conclusion The decreased expression of elastin and fibolin-5 was correlated with degree of POP, which indicated that elastin and fibulin-5 may play a role in the pathogenesis of POP.

To obtain the immunized mouse spleen cells, we immnnized the female BALB/C mice I.P. with schizonts or merozoites of the cultured blood forms of Plasmodium falciparum. The immunized spleen cells were fused with the SP2/0 myeloma cells, by which we obtained two strains of hybridoma secreting monoclonal antibodies against human Plasmodium falciparum. Using indirect immunofluorescence assay, we identified that the antibodies reacted only with the surface membrane antigens of the schizonts. These hybridomas were cultured continuously in vitro for over ten months and stably produced antibodies-IgG. The two hybridoma cell lines were designated as AEB2 and AGA4, and the number of their chromosomes was 98 and 100 respectively. The hybridoma cells were injected I. P. into the paraffin oil treated BALB/C mice to obtain the hybridoma ascitic fluid containing monoclonal antibodies. The ascitic fluid inhibited the growth of P. falciparum in vitro up to 89%. The inhibition test showed that antibodies were protective.

Objective To investigate the effects of tumor necrosis factor-alpha (TNF-α) on insulin sensitivity and glucose-lipid metabolism in TNF-α-induced IR mice. Methods Male C57BL/6J mice were given an intraperitoneal injection of TNF-α (H group,6μg/kg; M group,3μg/kg; L group,1μg/kg;twice daily) and saline (NC group) for 7 days. The plasma glucose and insulin were assayed during intravenous glucose tolerance test (IVGTT) and hyperinsulinemic-euglycemic clamp combined with 3-[3H] glucose as a tracer was carried out. Results After TNF-α treatment,fasting blood glucose (FBG),plasma insulin and free fatty acids (FFA) were significantly elevated in H group compared with NC,L and M groups (P<0.01 and P<0.05,respectively). There was a lower glucose tolerance in H group versus other three groups during IVGTT. The insulin release by glucose stimulation was higher in H group versus NC and L groups (P<0.01 and P<0.05). Basal glucose disappearance rate (GDR) and hepatic glucose production (HGP) were significantly increased in H group compared with NC group (P<0.01). During the steady-state of clamp,plasma insulin levels were significantly increased in H group versus NC group (341.7±17.7 vs 84.7±5.5mU/L,P<0.01). The suppressive effect of insulin on FFA was significantly blunted in H group compared with NC group (0.82±0.03 vs 0.43±0.07mmol/L,P<0.01). Steady-state glucose infusion rate (GIR) was significantly decreased in H group compared with NC group (39.1±2.3 vs 54.2±2.2 mg·kg-1·min-1,P<0.01). Although GDR was increased in both group,but it was still lower in H group than in control group(47.9±0.8 vs 53.9±2.0 mg.kg-1.min-1,P<0.01).As compared with baseline,HGP in the controls was almost completely suppressed during steady state of clamp,but in H group suppressed by approximately 41%. Conclusions High-dose TNF-α treatment induces the abnormality of glucose-lipid metabolism and the insulin resistance of hepatic and peripheral tissue in mice

Objective:To investigate the distribution of aldose reductase (AR) C-106T genetic polymorphism in Chinese Han population and its association with the risk for essential hypertension (EH).Methods:The AR C-106T polymorphism was genotyped in 148 Chinese EH patients and 137controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).The genotype distribution between groups was contrasted by x2- test and the degree of genetic association was evaluated by 95％ confidence interval (CI).Results:Frequency of the variant AR C-106T allele was 13.9％ (95％ CI:11.2％-16.6％) in the controls,which was significantly lower than that in the Japanese (18.4％ in 712 individuals,P=0.0063),the Australians (37.9％ in 240 individuals,P＜0.0001) and the Brazilians (34.7％ in 62individuals,P＜ 0.0001).The frequency ofAR C-106T allele was 11.7％ (95％ CI:7.9％-15.5％)in the EH patients.No significant difference in the allele frequency was observed between the EH patients and the controls (P=0.147).Conclusion:There is obvious racial difference in the distribution of AR C-106T polymorphism.The polymorphism is not associated with the risk for EH.

Objective To investigate the association of interferon (IFN) γ gene polymorphisms and risk and prognosis of HPV cervical infection.Methods PCR-ASP was used for detectiug IFN-γ rs2430561 polymorphism in 179 HPV positive patients and 328 HPV negative normal controls.Results The frequency of A allele of 63.7％ (228/358) was significantly higher than the frequency of T allele of 36.3％ (130/358) in HPV positive group (P =0.045).The frequencies were 41.3％ (74/179) in AA genotype and 14.0％ (25/179) in TT genotype,women carrying AA genotype increased the risk of HPV infection compare with those with TT genotype (OR =1.784,95％ CI:1.031-3.088,P =0.039).During follow-up,the rate of HPV positive again in AA genotype was 83.8％ (62/74),while TT genotype was 20.0％ (5/25).In the analysis of Kaplan-Meier,the cumulative HPV negative rates of AA,TA and TT genotype exhibited significantly different (P =0.008).The cumulative HPV negative rate of AA genotype was the lowest (1.1％-5.9％).Conclusions IFN-γ rs2430561 polymorphisms confer the susceptibility to HPV infection.Women with AA genotype exhibited higher risk of infection and inclined to be continuous status and recurrence after HPV infection.

Plasma obestatin level was determined in patients with impaired glucose regulation and type 2 diabetes mellitus.The plasma obestatin levels in patients of both groups were significantly decreased as compared with that in controls.Plasma obestatin level was negatively correlated with body mass index,HbA_(1C),waist-to-hip ratio,plasma insulin and HOMA-IR.Obestatin level seems to be related with metabolic disorder.