Objective To explore the role of leptin,COX-2 and p27 in bile-induced gastric mucosal injury in rats.Methods SD rats were divided into 4 groups: DGR(duodenogastric reflux) group,DGR+BDL(ligation of bile duct) group,BDL group and control group.The pathological changes of gastric mucosa and tight junction were observed,DIXON scores were warked out after 3 months.Immunohistochemistry was used to detect the expression of leptin,COX-2 and p27 in gastric mucosa with different lesions. Results The DIXON scores in DGR and DGR+BDL group were higher than those in BDL group and control group(P

Objective To observe the therapeutic effect of herbal decoction Kugan Yin for cystic hyperplasia of breast.Methods The 131 cases of cystic hyperplasia of breast in phlegm combining qi stagnation syndrome were randomized into treatment group(68 cases)which was treated by Kugan Yin and control group(63 cases)which was treated by Rupixiao Tablets(Tablets for dissolving breast nodules).The changes of symptoms and signs,level of serum sexual hormone and molybdenum target mammography analysis of two groups were observed for comparison.Results The total effective rate of treatment group was obviously better than that of con- trol group(P

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

OBJECTIVETo study the effect of Shenqi Fuzheng Injection (SQFZI) on erythrocyte immune function in patients after cardiopulmonary bypass (CPB).

METHODSTwenty patients ready for receiving CPB were randomly assigned to two groups, 10 in each group. Patients in the SQFZI group were treated with 250 mL SQFZI via intravenous dripping starting from 5 days before operation and after ending CPB once a day. Peripheral venous blood samples of all patients were collected at the time points of before anesthesia, and 6 h, 24 h, 72 h and 7 days after CPB, which were anticoagulated with heparin for dynamically detecting the changes of RBC-C3bRR and RBC-ICR.

RESULTSRBC-C3bRR and RBC-ICR in the SQFZI group were significantly higher at 6 h, 24 h, 72 h, and day 7 after operation than those in the control group respectively at the corresponding time points, and they restored to the normal levels at day 7, suggesting the erythrocyte immune function after CPB in the SQFZI group was superior to that in the control group (P < 0.05).

CONCLUSIONApplication of SQFZI in the peri-operative period can significantly improve the hypo-immunological function of erythrocyte caused by CPB, and promote the recovery of erythrocyte immunity.

Adult ; Aged ; Cardiopulmonary Bypass ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Erythrocytes ; drug effects ; immunology ; Female ; Heart Valve Diseases ; blood ; drug therapy ; surgery ; Heart Valve Prosthesis Implantation ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Phytotherapy ; Treatment Outcome

Objective To obtain a strain of glycoengineering yeast with higher N-glycosylation efficiency by overexpressing N-glycosyltransferase.Methods Through the selecting marker URA3 gene, a new glycoengineering yeast strain named 4-32-STT3D was constructed, which could overexpress the Leishmania major N-glycosyltransferase staurosporine and temperature sensitivity3 D subunit(STT3D) under the control of an inducible alcohol oxidase 1(AOX1) promoter.We analyzed the N-glycosylation status of anti-human epidermal growth factor receptor 2 ( HER2 ) antibody and granulocyte macrophage colony stimulating factor (GM-CSF) expressed in 4-32-STT3D using SDS-PAGE,Western blotting and peptide-N-asparigineamidase F(PNGase F).Finally the effect of STT3D on the growth rate of glycoengineering yeast was detected.Results SDS-PAGE showed that anti-HER2 antibody expressed in 4-32-HL had two components:the first one with a relative molecular mass 55 ×103 was glycosylated,while the second one with 50 ×103 was non-glycosylated,but anti-HER2 antibody expressed in 4-32-HL-STT3D had the component of 55 ×103 only without any non-glycosylated 50 ×103 .The above components became 50 ×103 with the digestion of PNGaseF.All of them proved to be antibodies by Western blotting.As a report protein,GM-CSF expressed in 4-32-GM-CSF had two components: 22 ×103 and 20 ×103, while in 4-32-GM-CSF-STT3D there was only one with 22 ×103 .All these components became 18 ×103 with the digestion of PNGase F.Statistical analysis showed that without induction,STT3D had no effect on the growth rate of glycoengineering yeast, while great effect was observed when STT3D was induced.Conclusion Glycoengineering yeast with the overexpression of N-glycosyltransferase has higher N-glycosylation efficiency.

Objective To investigate the improvement of ischemic hypoxic injury of brain after the transplantation of bone marrow mesenchymal stem cells (BMSCs).Methods Rats were randomly (random number) divided into sham operation group,cardiac arrest group and BMSCs treatment group (n =10 in each group).The model of cardiac arrest was induced by asphyxia.One hour after restoration of spontaneous circulation (ROSC),green fluorescent protein labeled BMSCs were transplanted via tail vein injection.At 3 and 7 days after transplantation,frozen sections of hippocampus was stained with hematoxylin-eosin (HE).The rest of brain tissue was weighed by an electronic balance.Brain water content (％) was calculated as (wet weight-dry weight) / wet weight × 100％.Results ①BMSCs were observed in hippocampus at 3 and 7 days after transplantation under fluorescent microscopy.②Compared with sham operation group and BMSCs treatment group,brain water content in cardiac arrest group was higher (all P ＜ 0.05).HE staining results showed that BMSCs transplantation could lessen hypoxia ischemia damage on brain.Conclusions BMSCs reduced the neurons damage induced by cardiac arrest and promoted neurological function recovery.

Background:As the empirical studies on human body are restricted extremely,the establishment and selection of suitable animal models are important for researches on ulcerative colitis( UC ). Aims:To compare the symptoms and colonic pathology of rat models with experimental colitis induced by dextran sulfate sodium( DSS ) and trinitrobenzene sulfonic acid( TNBS),so as to provide a reference for selecting animal models in UC-related studies. Methods:Drinking 4% DSS freely for 7 days or intrarectal administration of single dose 100 mg/kg TNBS-50% ethanol were used to establish experimental colitis model in Sprague-Dawley rats. The disease activity index( DAI)was assessed dynamically during the course of experiment. The whole colon was removed in batches for measurements of colonic damage score and activity of myeloperoxidase(MPO)at different time points. Results:The DAI score reached the peak at the 7th day and the 2nd day in DSS group and TNBS group,respectively,and decreased gradually afterwards. Six and one deaths occurred during the experimental course in DSS and TNBS groups,respectively. In DSS group,the duration of inflammation was short,the colonic injury was moderate and recovered after drug withdrawal. At the 18th day,the colonic damage score and MPO activity was 0. 25 ± 0. 50 and(0. 80 ± 0. 33)U/g,respectively,and no significant differences were seen between DSS group and normal control group. In TNBS group,the duration of inflammation was longer and the colonic injury was more severe. At the 21st day,the colonic damage score and MPO activity was 3. 60 ± 0. 55 and( 1. 60 ± 0. 39 ) U/g, respectively,and chronic inflammation was observed histologically. Conclusions:Both DSS and TNBS can induce experimental colitis model in rats. The course of TNBS-induced colitis model presents a transformation of acute to chronic inflammation,and may be more suitable for treatment-related studies of UC.

Objective Influence of cover statement and clarity of the messages upon risk decision making about thrombolysis therapy in patients with cerebral infarction was discussed in the Scenarios of thrombolytic therapy for acute cerebral infarction.Methods Inpatients and outpatients with cerebral infarction were randomly divided into two groups:detailed / simple cover statement group.Every participant underwent obscure messages and clear messages with only one frame:positive or negative frame.Results In the simple cover statement group,both obscure messages subgroup and clear messages subgroup were not influenced by the framing effect with preference to the risk seeking.In the detailed cover statement group,participants in the clear messages group were not influenced by the framing effect(x 2 =0.19,P＞0.05),while the subjects in the obscure messages group were actually affected by the framing effect,with the preference to risk seeking in the positive framework (x 2=7.90,P＜0.01).Conclusion The risk decision making about thrombolysis therapy in patients with cerebral infarction is influenced by cover statements and clarity of the messages.To improve the rate of thrombolysis therapy,the patients should be exposed to the obscure messages in the positive framing information under the detailed cover statement.

Objective To prepare endo-beta-N-acetylglucosaminidase H (Endo-H) expressed in Pichia pastoris, and apply it to N-glycosylation analysis .Methods One complete gene was synthesized on the basis of the cDNA sequence encoding Streptomyces plicatus reported in GenBank .The gene was cloned into the expression vector pPIC 9.The expression vector pPIC9-Endo-H was transformed into P.pastoris(JC308).The expression products were induced by methanol , puri-fied by two-step chromatography , used to analyze the glycan structures of RNaseB by the DNA sequencer assisted fluoro-phore-assisted carbohydrate electrophoresis (DSA-FACE)methods, and finally compared with peptide-N-asparagine amidase F(PNGase F).Results This enzyme expressed in P.pastoris(JC308) had the ability to hydrolyze natural or denatured high-mannose type of oligosaccharide linked by β-1,4-glycosidic bonds , but not complex-type oligosaccharide .The result of DSA-FACE showed that carbohydrate chains of Man 5 GlcNAc-Man9 GlcNAc could be obtained when RNaseB was hydrolyzed by Endo-H, and that Man5 GlcNAc2-Man9 GlcNAc2 chains became available when RNaseB was hydrolyzed by PNGase F . Conclusion Endo-H expressed in P.pastoris has bioactivity which can be used to analyze N-glycosylation with the method of DSA-FACE.

Objective To discuss the influence of framing information and time window information upon risk decision making about thrombolysis therapy in patients with cerebral infarction or their relatives in the scenarios of thrombolytic therapy for acute cerebral infarction.Methods Inpatients and outpatients with cerebral infarction or their relatives(n=327) were randomly divided into positive framing group(n=187) and negative framing group(n=140).Every participant underwent risky messages about thrombolytic treatment in three different time windows:0-90 minutes,91-180 minutes and 181-270 minutes.Results In any thrombolytic time window condition,there was no significant difference in thrombolytic tendency of participants between the positive framing group and the negative framing group (0-90 min:x2=1.24,P＞ 0.05;91-180 minutes:x2 =0.35,P＞0.05;181-270 minutes:x2 =0.07,P＞ 0.05).In a negative framework conditions,compared with the participants receiving thrombolytic information in 181-270 minutes time window,subjects receiving thrombolytic information in 0-90 minutes window were more tolerant of thrombolytic treatment (69.3 ％ vs 51.5 ％,P＜ 0.0125).Conclusion There are mutual effects of time window and framing information upon participants' preferences for thrombolysis.Different thrombolytic information are presented according to their corresponding time windows.