Objective To lower immunogenesis of human adult islets,reduce the dose of immunosuppressors and promote wide development of transplantation of adult islets,isolated adult islets were in vitro pretreated.Methods Isolated adult islets were cultured at 24 ℃ for 2 days(24 ℃ group) or pretreated using MHC-II monoclonal antibody(monoclonal antibody and complements were added and cultivated for another day following one-day-24 ℃-culture,antibody group) as observation groups.Adult islets were cultured at 37 ℃ in CMRL 1066 medium plus 20 % fetal bovine serum for 2 days as control.The immunogenics of the islets was detected using lymphocyte-islet mixed culture(MILC) and immunohistochemical staining of HLA-DR and lymphocyte common antigen(LCA).The function and activity of the islets were identified using()~3H-leucine incorporation assay,insulin release test and in situ apoptosis assay.Results Compared with control,MILC stimulation index was remarkably lower in the antibody group(P(0.05)).The percentage of HLA-DR and LCA positive cells in the antibody groupand in the 24 ℃ group was much lower than that in control(All P

Objective To explore the clinical efficacy of polyp of vocal cord. Methods The clinical data of 187 cases were retrospectively analyzed. Results According to the 187 cases, 164/187 cases were cured, 21 cases had obvious improvement, 2 cases have no effect, general effective rate is 98.9%. Conclusion Suspension laryngoscope were visual field widen and visual, every kind of surgical instruments binding equipments improve operation accuracy and reduce complication to acquire better clinical therapeutic effects.

Objective To explore the difference involved in the activation of inflammation pathway and the plasma level of inflammatory factors in the subjects with different sorts of insulin sensitivity. Methods The study was carried out in 38 women, consisting of obesity (n = 22 ) and control (n = 16 ) groups according to body mass index. The insulin sensitivity was assessed by homeostasis model assessment of insulin resistance (HOMAIR). Plasma concentrations of interleukin-6 (II-6) and IL-1β were determined by enzyme immunoassay. Western blot analysis was used to examine total protein expression and phosphorylation levels of IκB kinase (IKK) ,inhibitor of nuclear factor-κB ( IκB ) in peripheral blood leukcocytes. Electrophoretic mobility shift assay (EMSA)was used to detect the binding activity of NFκB. Results The levels of fasting plasma insulin[62.2 ( 20.0-127. 0) pmol/L vs 19. 15 ( 14. 2-47. 8 ) pmol/L, P<0. 01], HOMA-IR[2. 32 ( 0. 76-5.49 ) vs 0.70(0.53-1.7),P<0.0l], HbA1 C[(5.42±0. 45 ) % vs ( 5.08 ±0. 38) %, P<0. 05], triglyceride[( 1.75 ±0. 68 vs 1.22 ±0. 58 )mmol/L, P<0. 05], plasma IL-6[3. 15 (0. 03-22. 2) pg/ml vs 1.26 (0. 74-6.06 ) pg/ml, P<0. 01], and IL-1 β[6. 53 ( 0. 84-36 ) pg/ml vs 3. 16( 1.48-8. 86 ) pg/ml, P<0. 01]in obesity group were significantly higher than those in control group. Compared with control group, the levels of IKKo, IKKβ expression and IκBα serine phosphorylation in obesity group were markedly increased, while the expression of IκBα was significantly reduced. Accompanied with the degradation of IκBα protein, the binding activity of NFκB in obesity group was significantly increased. Conclusions The plasma levels of IL-6 and IL-1β were significantly raised in obesity group. The activation of IKK-IκB-NFκB pathway is closely associated with the genesis and development of insulin resistance in obese subjects.

To investigate the effects and the mechanism of visfatin on MIN6 cell signaling pathway and apoptosis induced by palmitate.Human recombinant visfatin promotes protein kinase B (Akt) and extracellularsignal regulating kinase (ERK)1/2 phosphorylation in dose-and time-dependent manner,and prevents MIN6 cell from apoptosis induced by palmitate (P<0.05 or P<0.01).The activation of Akt and ERK1/2 signaling pathway may be one of the molecular mechanisms of visfatin.

Visfatin was expressed in rat anti mouse islets,as well as in MIN6 cells. The visfatin expression was affected by various concentrations of environmental glucose (5.5 and 33.3 mmoL/L) and palmitate(0.5 mmol/L). As compared with low-level glucose medium (5.5 mmol/L, 1.0±0.11) , visfatin expression increased in media with high glucose and palmitate (1.32 ±0. 18, 1. 33±0. 15,1.72±0.27, all P<0. 05). The result suggests that visfatin seems to be involved in the regulation of insulin secretion.

Objective To investigate the effects of differentiated 3T3-L1 adipoeytes on inflammation of rat islet cells,as well as the protective effect of a-lipoic acid on the inflammation in vitro.Methotis Rat islet cells were divided into three groups:the control group,the experimental co-culture system group(cocuhured with differentiated mature 3T3-LI adipoeytes)and the intervention group (cocuhured with mature 3T3-LI adipocytes containing 4 μg/ml a-lipoic acid).Insulin releasing lest wag performed for estinmting the function of islet cells in culture supernatant of difierent groups.At the same time,the expression level of IKKIβin islet cells Was detected by western blot and realtime PCR.Results There was significant decrease of insulin stimulation index (SI) in experimental co-culture system group compared with the control group and intervention group(1.0 ±0.1 vs 2.6±0.2,2.5±0.5 respectively;P<0.01),while,the mRNA(4.62±0.60 vs1.00±0.46 and 2.25±0.75;P<0.01)and protein expression of IKKβ were significandy increased in the experimental group as compared with the other two groups.Conclusions In the co-culture system of adipocytes/islet cells,impaired function of islet cells could be induced by IKKβ activation,IKKβ Was a key molecule in inflamnmtion signal pathway in islet cells and could be activated by 3T3-LI adipocytes.a-lipoic acid Was able to reverse the impaired function of islet cells by suppressing IKKβ expression.

Objective To test whether glucagon like peptide-1 (GLP-1) would regulate the expression of visfatin in adipocytes,and to explore the mechanism of this effect.Methods Fully differentiated 3T3-L1 adipocytes were treated with GLP-1.Total RNA was extracted for analyzing the level of visfatin mRNA by quantitative RT-PCR.The media were collected for measuring the level of visfatin protein by enzyme linked immuno-assay (ELISA).In order to test the involvement of PKA pathway,the adipocytes were pretreated with a specific pharmacological PKA inhibitor H89 for 30 min before GLP-1 was added.Results GLP-1 increased visfatin expression in a time-and dose-dependent manner.The level of visfatin significantly increased at the concentration of 10 10 mol/L GLP-1 (P＜0.05),and reached the peak at 10-9 mol/L (P＜0.01).After incubation for 18 hours,GLP-1 dominantly increased the level of visfatin (P＜0.05).Inhibition of PKA pathway by H89 partially blocked the effect of GLP-1 on visfatin expression.Conclusions GLP-1 may enhance the expression of visfatin in 3T3-L1 adipocyte via the PKA pathway,which might contribute to the improvement in glucose homeostasis.

Heparinase III (HepIII) is a 73-kDa polysaccharide lyase (PL) that degrades the heparan sulfate (HS) polysaccharides at sulfate-rare regions, which are important co-factors for a vast array of functional distinct proteins including the well-characterized antithrombin and the FGF/FGFR signal transduction system. It functions in cleaving metazoan heparan sulfate (HS) and providing carbon, nitrogen and sulfate sources for host microorganisms. It has long been used to deduce the structure of HS and heparin motifs; however, the structure of its own is unknown. Here we report the crystal structure of the HepIII from Bacteroides thetaiotaomicron at a resolution of 1.6 Å. The overall architecture of HepIII belongs to the (α/α)₅ toroid subclass with an N-terminal toroid-like domain and a C-terminal β-sandwich domain. Analysis of this high-resolution structure allows us to identify a potential HS substrate binding site in a tunnel between the two domains. A tetrasaccharide substrate bound model suggests an elimination mechanism in the HS degradation. Asn260 and His464 neutralize the carboxylic group, whereas Tyr314 serves both as a general base in C-5 proton abstraction, and a general acid in a proton donation to reconstitute the terminal hydroxyl group, respectively. The structure of HepIII and the proposed reaction model provide a molecular basis for its potential practical utilization and the mechanism of its eliminative degradation for HS polysaccarides.
Amino Acid Sequence ; Bacteroides ; enzymology ; Catalytic Domain ; Crystallography, X-Ray ; Heparitin Sulfate ; metabolism ; Kinetics ; Models, Molecular ; Molecular Sequence Data ; Polysaccharide-Lyases ; chemistry ; metabolism ; Substrate Specificity

Objective To explore the levels of serum adiponectin (ADPN) and retinol-binding protein 4 (RBP4) in patients with type 2 diabetes mellitus (T2DM) and obstructive sleep apnea-hypopnea syndrome (OSAHS).Methods A total of 155 patients with T2DM and OSAHS were selected and divided into mild group (n=62),moderate group (n=53) and severe group (n=40) according to apnea-hypopnea index (AHI),35 OSAHS patients were designed as OSAHS group,42 T2DM patients were selected as T2DM group,and 38 healthy controls were selected as control group.Serum ADPN and RBP4 levels were detected by ELISA.Results Compared with healthy controls,the serum level of RBP4 significantly increased and the ADPN level significantly decreased in the other three groups (P<0.01).In subgroups of OSAHS and T2DM group,the level of RBP4 increased with increasing of AHI level,ADPN level is on the contrary (P<0.01).Conclusion The serum levels of ADPN and RBP4 are closely related with the severity of T2DM patients with OSAHS.

Objective To explore the levels of serum adiponectin (ADPN) and retinol-binding protein 4 (RBP4) in patients with type 2 diabetes mellitus (T2DM) and obstructive sleep apnea-hypopnea syndrome (OSAHS).Methods A total of 155 patients with T2DM and OSAHS were selected and divided into mild group (n=62),moderate group (n=53) and severe group (n=40) according to apnea-hypopnea index (AHI),35 OSAHS patients were designed as OSAHS group,42 T2DM patients were selected as T2DM group,and 38 healthy controls were selected as control group.Serum ADPN and RBP4 levels were detected by ELISA.Results Compared with healthy controls,the serum level of RBP4 significantly increased and the ADPN level significantly decreased in the other three groups (P<0.01).In subgroups of OSAHS and T2DM group,the level of RBP4 increased with increasing of AHI level,ADPN level is on the contrary (P<0.01).Conclusion The serum levels of ADPN and RBP4 are closely related with the severity of T2DM patients with OSAHS.