Objective:To study the expressions of Vascular Cell Adhesion Molecule 1 (VCAM-1) and Intercellular Adhesion Molecule 1 (iCAM-1) in patients with endometriosis and to explore its effect on the onset of endometriosis.Methods:VCAM-1 mRNA and ICAM-1 mRNA were detected by real-time PCR method in 15 specimens of eutopic endometrium and 15 specimens of ectopic endometrium from patients with endometriosis(EMT group)and 15 specimens of endometrium from patients without endometriosis(control group).VCAM-1 and ICAM-1 protein were detected by Western-Blot method.Soluble VCAM-1 and soluble ICAM-1 in 44 serum of patients with endometriosis(EMT group) and 28 serum of patients without endometriosis(control group) were tested by ELISA.Meanwhile the correlation of expressions of VCAM-1 and ICAM-1 were analyzed in different groups.Results:①The expressions of VCAM-1 mRNA and ICAM-1 mRNA in ectopic endometrium were significantly higher than those of eutopic endometrium and the controls(P ＜0.01).There was no significant different mRNA expression between eutopic group and control group(P＞0.05).②The protein expressions of VCAM-1 and ICAM-1 in eutopic group were significantly higher than those of ectopic group and the control group(P ＜0.01).The protein expression of VCAM-1 in eutopic group were higher than those in the controls group(P＜0.01).There was no significant different protein expression of ICAM-1 between eutopic group and the controls group(P ＞0.05).③The concentration of sVCAM-1 and sICAM-1 in serum from patients with EMT were significantly higher than those in the control group(P＜0.01).The concentration of slCAM-1 in serum from patients with EMT in stage Ⅲ-Ⅳ were significantly higher than those in stage Ⅰ-Ⅱ (P ＜ 0.01).④There was no significant correlation in the expression of mRNA,protein and serum between VCAM 1 and ICAM-1 in ectopic group and eutopic group(P ＞ 0.05).Conclusions:The abnormal expressions of VCAM-1 and ICAM-1 in ectopic endometrium may play an important role in the development of endometriosis.The signaling pathways and protein expression way may be different between VCAM 1 and ICAM 1 in endometriosis.

Objective To study a functional variable fragment of heavy chain(VH)antibody against the terminal protein(TP)region of hepatitis B virus(HBV)polymerase introduced by human immunodeficiency virus Tat protein transduction domain(TAT)and the inhibitive activity of TAT-VH on the replication of HBV in vitro.Methods The gene encoding TAT-VH was cloned into prokaryotic expression vector pET28a(+).Recombinant plasmid was transduced into E coli BL21(DE3)LysS,then the protein was expressed and purified.The purified TAT-VH fusion protein was added into HepG2.2.15 cell culture.The transduction efficiency was evaluated by indirect fluorescence assay(IFA).The cytotoxicity of TAT-VH was detected by Methabenzthiazuron(MTT)assay.HBV DNA level in HepG2.2.15 cell culture was measured using quantitative polymerase chain reaction(PCR).The data were analyzed by one-factor analysis of variance and t test.Results TAT-VH fusion protein was successfully expressed and purified.It was confirmed by IFA and MTT assay that TAT-VH was introduced into HepG2.2.15 cells and the cell growth was not affected.The level of HBV DNA in supernatant of HeDG2.2.15 cell culture with 5 000 nmol/L TAT-VH was(1.211±0.132)lg copy/mL,which was significantly lower than control group[(5.325±0.041)lg copy/mL,t=72.91,P＜0.05].Meanwhile,the level of intracellular HBV DNA was(3.521±0.411)lg copy/mL,which was significantly lower than control group[(8.532±0.132)lg copy/mL.t=28.41,P＜0.05].Conclusion The HBV replication is inhibited by anti-TP TAT-VH antibodies in vitro,which provides valuable experimemal basis for developing therapy of HBV infection with intracellular antibody.

Objective:To isolate the variable fragments of heavy chain(VH) against the terminal protein(TP) region of hepatitis B virus(HBV) DNA polymerase(Pol) with protein fragment complementation assay(PCA).Methods:The TP region of HBV secreted by the HepG2.2.15 cells was used as an antigen,and the antibodies were selected with PCA.In this assay,two interacting proteins(target and antibody) are genetically fused to the two halves of the dissected enzyme dihydrofolate reductase.Binding of the two partners reassembles this enzyme and reconstitutes its activity,thus allowing growth on minimal mediem.Results:There were three TP region antigen-specific VHs could be directly in vivo selected with PCA.Sequence analysis showed that each TP-specific VH had a different sequence.Conclusion:Our results show that TP region antigen-specific VH could be directly in vivo selected with PCA.This system were powerful as a routine system for generating antibodies,especially in functional genomics.

Objective To investigate the possible pathogenesis of the cognitive function in unilateral frontal bottom laceration by follow-up study in patients after one month of the onset. Methods MMSE, Loewenstein Occupational Therapy Cognitive Assessment (LOTCA), Montreal Cognitive Assessment (MoCA), Wisconsin Card Sorting Test (WCST) scales were used to evaluate neurocognitie function in 42 patients after one month of onset of unilateral frontal bottom laceration and 45 normal controls. The wave amplitude and the latency of the endogenous composition N2, P3 of P300 were measured at the cognitive potential instrument. Level of AChE was determined by ELISA and active AChE was analyzed by the ration analyses. Stepwise multivariate regression analyzed the correlation of the overall cognitive function and the lever and active of AChE. Results The cognitive test scores in patients were significantly worse than those in normal controls. The ability of recite sentences, fluency of words, reading, understanding language,cognitive transfering decreases in the left frontal bottom laceration patients (Group A, 23 cases), while the ability of attention, action, organization, graphics depicting, abstract epitoming, logical thinking were all seriously impaired in the patients with right frontal bottom laceration (Group B, 19 cases). The latency of the endogenous composition N2, P3 in patients ( Group A: (322. 4 ± 17.0), (410. 1 ± 19.9) ms; Group B:( 308.4 ± 15.6), (385.5 ± 17.4) ms) is more lengthen ( F = 4. 084, P = 0. 018; F = 3.467, P = 0. 038 )than the normal controls ( (268.6 ± 14. 7 ), ( 369. 2 ± 15. 4 ) ms) and the wave amplitude is lower ( F =2. 986 ,P =0. 047 ;F =3. 313 ,P =0. 041 ). The latency of N2 ,P3 in Group A of is more lengthen than Group B, while the wave amplitude is higher. The difference of the active of AChE in patients and control groups had no statistical significance, however, the level of AChE in two groups had statistical significance. The comparison of the active and the total AChE in patients has also not statistical significance. The correlation of the overall cognitive function has the linear regression with the parts of the brain and the level of AChE ( rY1.2 = 0. 584, P = 0. 039; rY2.1 = 0. 726, P = 0. 017 ). The standardized regression coefficients showed the level of AChE has the biggest influence to the overall cognitive function ( |Beta| =0. 3601, rY2.1 =0. 726).Conclusions AChE may be one of the important factors in the cognitive function after frontal bottom laceration. The specific damages of cognitive function in unilateral frontal bottom laceration patients closely relate with the lesion locations in the injured frontal bottom laceration.

Objective To observe the curative effect of methylprednisolone combined with entecavir in treatment of hepatitis B virus (HBV) related early stage liver failure.Methods One hundred and twenty-six patients with HBV related early stage liver failure were divided into treatment group (68 cases) and control group (58 cases) by random digits table method.The patients in 2 groups were given conventional hepatinica treatment and entecavir antiviral treatment,but the patients in treatment group were added methylprednisolone and pantoprazole.The alanine aminotransferase (ALT),total bilirubin (TBil),albumin,prothrombin time (PT),HBV-DNA,tumor necrosis factor (TNF)-α,interleukin (IL)-6 levels were compared between 2 groups,and the adverse reaction of methylprednisolone was observed.Results The ALT,TBil,PT and albumin levels after the first,second,fourth,sixth and eighth week of treatment in treatment group were significantly better than those in control group,and there were statistical differences (P ＜ 0.05).There was no statistical difference in HBV-DNA between 2 groups (P ＞ 0.05).The TNF-α and IL-6 levels after the first and second week of treatment in treatment group were (4.13 ± 1.25) and (1.98 ± 0.67) p g/L,(3.21 ± 0.75)and (1.23 ± 0.29) μ g/L,and in control groups were (5.89 ± 1.78) and (3.67 ± 0.87)μ g/L,(4.12 ± 0.88) and (2.68 ± 0.81) μ g/L.The TNF-α and IL-6 levels in treatment group were significantly lower than those in control group,and there were statistical differences (P ＜ 0.05).The effective rate in treatment group (79.41％,54168) was significantly higher than that in control group (51.72％,30/58),the fatality rate in treatment group (2.94％,2/68) was significantly lower than that in control group (24.14％,14/58),and there were statistical differences (P ＜ 0.05).The adverse reaction of methylprednisolone in treatment group was not found.Conclusion The methylprednisolone combined with entecavir can improve liver function and survival rate in patients with HBV related early stage liver failure,and adverse reaction of methylprednisolone is rare.

Four-week-old male Sprague-Dawley rats were rendered diabetic by intraperitoneal injection of streptozotocin (STZ) after feeding high-fat-diet for 8 weeks,and divided into diabetes group and tumor necrosis factorrelated apoptosis ligand(TRAIL) group.Normal rats severed as a control group.Treatment with TRAIL lasted for 3 months.Total cholesterol,triglycerides,low-density lipoprotein-cholesterol,blood glucose,and insulin levels were decreased in TRAIL group,as compared with diabetes group.Area of atherosclerotic lesion in TRAIL group [(23.8 ± 5.7) ％] was significantly smaller than that in diabetes group [(47.6 ± 7.8) ％].It suggested that TRAIL may reduce the area of atherosclerotic lesion in diabetic rats.

Objective To investigate the effects and mechanisms of valproic acid on brain edema,neurobehavioral outcome and inflammatory response after traumatic brain injury (TBI) in rats.Methods TBI animal models were established using Feeney's method.Fifty-four SD male rats,weighting 220-250 g,were randomly divided into 3 groups (n =18):sham operation group (group sham),traumatic brain injury group (group TBI) and valproic acid treatment group (group TBI + VPA).Experimental rats were treated with valproic acid (300 mg/kg,twice daily) by intraperitoneal injection.Rat behavioral outcomes were measured by modified neurologic severity score (mNSS) tests at day 1,3,and 7 after TBI.Brain water content was measured with wet-dry weight method.The blood cells infiltration into cerebral cortex were tested with immunohistochemistry staining against ED-1 for macrophage.Inflammatory cytokines (INF-γ,tumor necrosis factor-α,interleukin-6) were measured by Western blotting.The statistical analysis were performed by ANOVA and chi-square tests using the statistical software program SPSS 13.0.Results Compared with the Sham group,the levels of brain edema,mNSS and macrophage cell infiltration were significantly increased after TBI (all P =0.00).The expressions of inflammatory cytokines were also increased significantly (all P =0.00).Compared with the TBI group,TBI + VAP group had significantly lower brain water content[3day:(80.12 ±0.59)％ vs.(82.14 ±0.67)％,P=0.04;7day:(74.74 ±0.72)％ vs.(77.93 ±0.48)％,P=0.01],and mNSS scores [3 day:(10.53 ±0.32) vs.(11.74 ±0.48),P =0.02;7 day:(7.97 ± 0.32) vs.(10.73 ± 0.42),P =0.01].VPA suppressed macrophage cell infiltration into cerebral cortex [(36.44 ± 0.72) ％ vs.(25.93 ± 0.48) ％ P =0.00].Meanwhile,VPA inhibited the expressions of inflammatory cytokines (INF-γ,TNF-α,IL-6) (P ＜ 0.05).Conclusions Treatment with VPA markedly reduced brain edema and improved neurological outcomes after TBI,possibly mediated by inhibited TBI-induced cerebral inflammatory responses and macrophage cell infiltrating into cerebral cortex.

Objective: To explore the effect of irisin on atherosclerosis with possible mechanisms in diabetes mellitus (DM) mice. Methods: A total of 30 ApoE-/- mice were randomly divided into 2 groups: Control group, the mice received citrate buffer solution for modeling control,n=10. DM group, the mice received streptozotocin injection for DM modeling,n=20; the DM group was further divided into 2 subgroups as DM control (DM-C) group, the mice received normal saline injection for 12 weeks and DM + irisin group, the diabetic mice received irisin injection for 12 weeks.n=10 in each subgroup. With 4 weeks of irisin intervention, the endothelium-dependent vasodilatation was detected. With 12 weeks of intervention, the blood levels of tumor necrosis factor-α (TNF-α), high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6) and oxidized low-density lipoprotein (ox-LDL) were examined by ELISA, the plaque areas in aortic en face and cross sections were measured by Oil red O or HE staining, the macrophages/T lymphocytes inifltration in plaques were detected with immunohistochemistry, and the mRNA expressions of IL-6, IL-10, TNF-α were determined by RT-PCR. Results: Compared with DM-C group, DM + irisin group presented improved endothelium-dependent vasodilatation, decreased levels of blood inlfammatory factors, reduced plaque on face area sections (22.57 ± 2.17) % vs (35.09 ± 2.38) % and cross sections (19.36 ± 1.85) % vs (25.53 ± 7.87) %,P < 0.05, less macrophages (30.5 ± 2.79) % vs (41.34 ± 9.13) % T and lymphocytes infiltration (28.11 ± 4.24) % vs (35.79 ± 9.11) % in plaques and lower mRNA expressions of inflammatory factors(IL-6: 1.76 ± 0.50 vs 3.78 ± 1.15; TNF-α: 1.05 ± 0.30 vs 2.11 ± 0.48; ICAM-1: 1.96 ± 0.69 vs 2.71 ± 0.72; VCAM-1: 0.87 ± 0.21vs 1.45±0.25; MCP-1: 1.34 ± 0.34 vs 1.77 ± 0.55) at aortic wall, P<0.05.Conclusion: Irisin may improve atherosclerosis condition in ApoE-/- DM mice, the endothelial protection and antiinflammatoryreaction were the important mechanisms. Irisin has the potential for preventing/treating atherosclerosis.

Objective To study the association between transforming growth factor-β1 (TGF-β1) polymorphism and type 2 diabetes mellitus in Han population of Shanghai. Methods In this case-control study , 1 234 cases of T2DM patients were recruited and 1 272 healthy individuals were selected as control. Five ml of blood sample was collected from each subject ,from which the whole genomic DNA was extracted.The polymorphism was detected by the Taqman technology. Result Significant association was observed in TGF-β1 T896C genotypes and alleles with T2DM (P = 0.0001 and P = 0.004, OR = 1.18 [1.05 ~ 1.33], respectively). Conclusion The polymorphism of T896C in TGF-β1 gene may be associated with T2DM in Han population from Shanghai.

Objective To examine the clinical features of sporadic patients with H7N9 avian influenzain Taizhou city of Zhejiang province and to characterize its viral genes.Methods Fifteen patients with H7N9 influenza infection confirmed by Zhejiang Provincial Centre for Disease Control and Prevention during January 201 4 and January 201 5 were included in the study.The basic diseases,poultry exposure history,clinical manifestation,laboratory examination,imaging features,treatment and outcome and viral gene sequencing were analyzed retrospectively.Results The first clinical symptoms were fever and cough in all patients,acuterespiratory distress syndrome(ARDS)occurred in 1 3 patients,the average time from onset to antiviral therapy was (7 ±2)d.Among 1 5 patients,9 survived and 6 died,including 2 died of multiple organ failure (MOF).The phylogenetic tree showed that there was highly homologous in hemagglutinin (HA)and neuraminidase(NA)genes between human H7N9 virus strains and poultry reference strains.The result of genetic sequencing indicated that human H7N9 virus strains had mutations at 226 (Q226L)sites in HA protein.Conclusions ARDS is likely to occur in patients with H7N9 viral infection,and early antiviral treatment usually leads to a good prognosis.With the occurrence of adaptive mutation in avian influenza virus H7N9,spread from poultry to the human beings may take place.