Glucagon-like peptide-1 is one of the most important members of incretin, which has an unusual therapeutic potential in type 2 diabetes mellitus and obesity. Its analogues are resistant to dipeptidyl peptidase Ⅳ hydrolysis and begin to be used clinically. These preparations may have important application perspective as new therapeutic agents in the treatment of diabetes mellitus and obesity.

Objective The monoclone antibody against insulin was prepared and used to measure plasma level of insulin in patients with type 2 diabetes mellitus. Methods Balb/c mice were immunedwith cross-linking insulin-BSA as immunogen. Four monoelone cell strains stably secreting insulin monoclone antibody were prepared by applying hybridoma technique. The valence, subclass and affini-ty constant of the monoclone antibody were analyzed. The serum insulin content was measured with competitive inhibition ELISA in 90 patients with type 2 diabetes mellitus and 90 healthy controls. Re-suits The affinity constant of the established monoclone antibody reached to 1011. The subclass of 1 strain was Igab(k),and the other 3 strains were Ig1(k) srbclass.The mean level of insulin was significantly lower in patients with type 2 diabetes mellitus than that of healthy controls. Conclusion These results implicated that the insulin insufficient might be due to the depletion of the pancreatic be-ta cell function of patients with type 2 diabetes mellitus, and the pathogenetic condition might be alle-viated by supplying insulin. About 10% patients are with higher insulin level than healthy controls,which may be due to insulin resistance. It is inadvisable to treat these patients with insulin therapy.

ObjectiveTo investigate the effects and mechanisms of IL-32γ-shRNA-mediated gene silencing on the proliferation and apoptosis of fibroblast-like synoviocytes in patients with rheumatoid arthritis (RA).MethodsA eukaryotic expression plasmid of shRNA targeting IL-32γ was transfected into fibroblastlike synoviocytes by liposome in patients with rheumatoid arthritis.RT-PCR was used to determine the expression level of IL-32γ.Western blotting was used to detect the levels of cyclin D1 and p-Akt.The proliferation of RA-FLS was examined by MTT.Cell cycles were analyzed by flow-cytometry.The apoptosis of cells were measured by TUNNEL.Comparisons between groups were tested by t test.Results ① The expression of IL-32γwas significantly inhibited by shRNA-IL-32γ-expressing plamid PGCsi 3.0 targeting sequence 1,2 and 3,and the inhibition rate had reached 75.6％,66.2％ and 64.1％,respectively.② The absorbance value of proliferation of RA-FLS in EASY-shRNA-IL-32γ group was significantly lower than that in the shRNA-control group and normal group at day 3 [(0.23±0.03) vs (0.35±0.03) and (0.36±0.04),P＜0.05] and 5 [(0.27±0.03) vs (0.52±0.05) and (0.53±0.04),P＜0.01 ] after transfection.③ The rate of RA-FLS at phase G1 in the EASY-shRNA-IL-32γ group was significantly higher than that in the shRNA-control group and normal group respectively [(88±6)％ vs (69±5)％ and (68±4)％,P＜0.05],while those at phase S+G2 in the EASY-shRNA-IL-32γgroup was significantly lower than that in the shRNA-control group and normal group [ ( 13.6±3.0)％ vs (30.2±4.1)％ and(32.1±4.3)％,P＜0.01].④The rate of RA-FLS apoptosis in the EASY-shRNA-IL-32γ group was significantly higher than that in the shRNA-control group and normal group[(20.50±3.21 )％ vs (9.20±0.32)％ and (8.60±0.22)％,P＜0.01].⑤ The expression of cyclin D1(0.36±0.04) and p-Akt(0.31±0.03) in the EASY-shRNA-IL-32γ group was significantly lower than that in the shRNA-control group [ (0.59±0.08) and (0.53±0.06)] and normal group [(0.61±0.07) and (0.52±0.06),P＜0.01].ConclusionEASYshRNA-IL-32γ can inhibit RA-FLS proliferation by down-regulating the expression of cyclin D1 and induce RA-FLS apoptosis by down-regulating the expression of p-Akt.

[ ABSTRACT] AIM: To investigate the effect of chronic intermittent hypoxia on AMP-activated protein kinase ( AMPK) pathway in the brain of young rats.METHODS:Part one:SD mice (3~4 weeks old) were randomly divided into 4 groups (n=8): simulated air control group for 2 weeks (2AC), chronic intermittent hypoxia group for 2 weeks (2IH), simulated air control group for 4 weeks (4AC) and chronic intermittent hypoxia group for 4 weeks (4IH).Part two:SD mice (3~4 weeks old) were randomly divided into 2 groups (n=8): chronic intermittent hypoxia group for 4 weeks (4IH) and chronic intermittent hypoxia group treated with AMPK inhibitor for 4 weeks (4IHI).After modeling, the eight-arm maze test was performed.TUNEL method was used to detect the neuronal apoptosis in the hippocampal and pre-frontal cortical tissues.The mRNA expression of adenosine A2a receptor was examined by RT-qPCR, and the protein levels of phosphorylated AMPK (p-AMPK) and mammalian target of rapamycin (p-mTOR) were determined by Western blot.
RESULTS:Compared with control group, the numbers of reference memory error ( RME) , working memory error ( WME) and total error (TE) in 2IH group and 4IH group significantly increased (P<0.01).Compared with 2IH group, the num-bers of errors in 4IH group also increased significantly (P<0.01).Compared with 4IH group, the values in 4IHI group significantly decreased.Compared with control group, the neuronal apoptosis of hippocampus and prefrontal cortex in 2IH group and 4IH group increased, and that in 4IH group was more evident (P<0.05).In 4IHI group, the neuronal apopto-sis decreased.The mRNA expression of adenosine A2a receptor in the hippocampal and cortical tissues in 2IH group and 4IH group was higher than that in control group.The protein level of p-AMPK was higher, and p-mTOR was lower in 2IH group and 4IH group, and those in 4IH group were more evident (P<0.05).Compared with 4IH group, the protein level of p-AMPK was lower, and p-mTOR was higher in 4IHI group.CONCLUSION: Chronic intermittent hypoxia induces neuronal apoptosis, resulting in impairment of learning and memory in a time-dependent manner by upregulating adenosine A2a receptor, activating AMPK activity, and inhibiting mTOR phosphorylation in rats.

Objective To study the feasibility and safety of measles vaccination in women of childbearing age and to understand the influence of measles vaccination on the fetal transmission measles antibody level of the infants.Methods From January 1,2012 to December 31,2012,600 women of childbearing age were included in this study.The measles IgG antibody was detected,then all participants were randomized according to the IgG level.The measles IgG antibody of participants in non-vaccinated group was detected predelivery in hospital by enzyme linked-immunosorbent assay (ELISA).That of participants in vaccinated group was detected 3 months after vaccination with measles mumps rubella combined vaccine and predelivery by ELISA.And measles nuclear protein fragment gene (measles virus nucleoprotein,MVN) in the blood was detected by reverse transcription polymerase chain reaction (RT PCR).A total of 368 participants gave birth within 2 years after vaccination,and 357 infants 8 months after birth were healthy.In non-vaccinated group,8 infants were diagnosed with measles within 8 months after birth.Finally,349 8-month infants were enrolled in the study,including 52 whose mothers in high antibody without vaccination group,65 whose mothers in high antibody with vaccination group,110 whose mothers in low antibody without vaccination group and 122 whose mothers in low antibody with vaccination group.The measles IgG antibody levels in the blood of all the 8-month infants were detected.Data were analyzed using t test,one-way ANOVA (Newman-Keuls was used for comparison between groups) and Pearson analysis.Results The measles IgG antibody level of expectant women in high antibody without vaccination group was significantly lower compared to that in high antibody with vaccination group ([268.5±74.9] IU/mL vs [578.3t208.1] IU/mL,Q=15.57,P＜0.01).That in low antibody without vaccination group was also significantly lower than low antibody with vaccination group ([169.4+42.3] IU/mL vs [584.7+195.8] IU/mL,Q=29.54,P＜0.01).The results of MVN RT-PCR after 3 months of vaccination showed no positive bands in all blood samples.Two of the expectant women in high antibody without vaccination group and one in low antibody without vaccination group were positive for MVN bands.Among 8-month infants,the levels of antibody in high antibody without vaccination group and high antibody with vaccination group were (106.3 ± 36.8) IU/mL and (291.8±86.5) IU/mL,respectively,which was statistically different (t=23.33,P＜0.01).Those in low antibody without vaccination group and low antibody with vaccination group were (87.1 ± 26.4) IU/mL and (274.0±72.5) IU/mL,respectively,which was statistically different (t =33.27,P＜0.01).The measles antibody level of expectant women was positively correlated with their 8-month infants (r=0.652,P＜0.01).All 8 infants who were diagnosed with measles were delivered by women without vaccination,and the measles infection rate of infants was significantly different between women with and without vaccination (P=0.002).Condusion It is feasible for women of childbearing age to receive measles vaccination,which can increase the measles IgG antibody level of both expectant women and their infants.

Objective: To investigate the relationship between the EBV-induced liver injury and caspase-3/6 in children. Methods: Data of 249 patients seen from July 2016 to June 2017 who got infection with EBV were collected in second affiliated hospital of Wenzhou Medical University and the patients were divided into two groups; 168 patients who were diagnosed with hepatitis were selected for abnormal liver function group, laboratory tests were performed in acute phase and convalescent phase. Meanwhile the 81 patients, whose liver function were normal were selected for normal liver function group. Two ml of blood plasma was collected from each patient from both groups at the beginning and after a week’s treatment of the abnormal liver function group. The patients were aged from 1 to 14 years. The abnormal liver function group was further divided into four groups: young children (1-3 years old), preschool children (4-6 years old), school children (7-10 years old), teenagers (10-14 years old). Firstly, we recorded their alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), gamma glutamyl transpeptidase (GGT) and direct bilirubin (DBIL), then by the use of enzyme linked immunosorbent assay (ELISA) we measured the protein level of caspase-3 and caspase-6. Results: In the group of abnormal liver function group, in the acute phase of the teenagers the level of ALT, caspase-3 and caspase-6 are improved significantly(927.2±82.5 vs. 158.5±41.4, P<0.001, 169.8±52.8 vs. 91.5±10.1, P<0.001, P<0.05, 82.2± 2.6, P<0.001, 965.8±31.5, P<0.001, 184.8±14.3, P<0.001), while the AST, TBLL, GGT and DBLL had no significant difference. Conclusions In the process of EBV-induced liver injury, there was a linear correlation between the increase of caspase-3/6 and ALT in the acute stage, suggesting that EBV-induced liver injury is closely related to caspase-3/6.