Objective To evaluate the role of O-GlcNAc protein modification in attenuation of brain damage by glutamine in septic rats.Methods Sixty male SD rata weighing 180-240 g were randomly divided into 4 groups:sham operation group(group S,n =12),sepsis group(group CLP,n =16),glutamine group(group G,n =16),an inhibitor of O-linked-N-acetyl glucosamine transferase Alloxan + glutamine group(group G + A,n =16).Rats were submitted to sepsis by cecal ligation and perforation(CLP).Glutamine(Gln)0.75 g/kg was injected iv after CLP in group G.Gln 0.75 g/kg was injected iv and Alloxan 90 mg/kg was injected ip after CLP in group G + A.Equal volume of normal saline was given in group S and group CLP.A1 24 h afler CLP,the neural reflex score was evaluated,then rat was sacrificed.The brain was removed for measurement of brain water content,observation of histopathology and determination of O-GlcNAc-modified protein expression.Results Compared with group S,neural reflex score and brain water content were significantly increased in groups CLP,G and G + A(P ＜ 0.05).Compared with group CLP,neural reflex score and brain water content were significantly decreased in groups G and G + A(P ＜ 0.05),and the expression of O-GlcNAc-modified protein was upregulated in group G(P ＜ 0.05),Compared with groups G,neural reflex score and brain water content were significantly increased,and the expression of O-GlcNAc-modified protein downregulated in group G + A(P ＜ 0.05).There was no significant difference in O-GlcNAc-modified protein expression among groups S,CLP and G + A.Conclusion Glutamine attenuates brain damage through O-GlcNAc protein modification in septic rats.

Serum B7-H4 protein levels were measured by enzyme-link immunosorbent assay (ELISA) in 62 patients with epithelial ovarian cancer, 20 patients with benign ovarian diseases, and 30healthy subjects.Serum B7-H4 protein levels in epithelial ovarian cancer, benign ovarian diseases and normal controls were 0.72(0.32,1.88 )μg/L, (0.45±0.21)μg/L,(0.38 ± 0.19 ) μg/L respectively (P＜0.01).The serum level of B7-H4 protein in epithelial ovarian carcinoma was correlated with FIGO stages, lymph node metastasis, histological grade and histological types ( all P ＜ 0.01 ), but not with menopause and ascites (P＞0.05).The receiver operating characteristics curve analysis revealed that B7-H4protein had a sensitivity and specificity of 69% (43/62)and 86%(43/50) for diagnosis of epithelial ovarian cancer.

Objective To investigate the change and clinical significance of the serum level of human epididymis secretory protein 4 (HE4) in elderly patients with epithelial ovarian cancer (EOC).Methods The serum levels of HE4 were determined by ELISA in EOC patients (n= 33), benign ovarian disease patients ( n = 17) and healthy women ( n = 20), all of them aged 60 years or over. And there were other 31 EOC patients and 20 healthy women aged less than 60 years at the same period as controls. Results In healthy people, the serum level of HE4 was higher in the group with age ≥60 years than with age ＜60 years [(32.25± 13. 15) pmol/L vs. (24.59±8.60) pmol/L]. The serum HE4 level had a positive correlation with age (r=0. 525, P＜0.01). The median level of HE4 was significantly higher in elderly EOC patients than in benign ovarian disease patients (81.50 pmol/L vs.45.60 pmol L, U=168.5, P＜0.05) and healthy women (81.50 pmol L vs. 33.30 pmol L, U=76.5, P＜0. 01). The HE4 had a significantly higher area under ROC curve when compared with CA125 (0. 799 vs. 0. 782), while the cut-off value was set 65.43 pmol L with a sensibility and specificity of 66.67% and 86.49% respectively. The diagnostic efficacy of HE4 was higher than CA125. There was no significantly statistical difference between elderly and non-elderly EOC patients in serum HE4 levels (U= 101.2, P＞0. 05). Conclusions Age is an important factor which can affect human serum level of HE4. HE4 determination is helpful in the diagnosis of ovarian epithelial carcinoma in the elderly patients, and the combined detection of HE4 and CA125 is an useful adjuvant diagnostic measure.

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.