The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21-CodonPlus (DE3)-RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM<'4B> by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD<,tested serum>/OD<,negative serum>≥2.1- The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.

Objective To investigate the changes of gastrin(GAS) and somatostatin(SS) in diabetic gastroparesis (DGP). Methods Totally 103 patients were divided into normal control group (n =17),simple gastroparesis(GP) group (n =30),simple diabetes mellitus(DM) group (n =23) and diabetes mellitus complicated with gastroparesis (DGP) group (n =33). Radioimmunity and RT-PCR were performed to detect the serum GAS and SS levels and the mRNA expression in the gastric mucosa tissue of 4 groups. Results There were significant differences in the serum levels and protein expression of GAS and SS between simple gastroparesis group,simple diabetes mellitus group, DGP group and the normal control group(P< 0.05). The differences between DGP group and normal control group was the most significant (P< 0.01), the differences between DGP group and the simple gastroparesis group or simple diabetes mellitus group were significant (P< 0.05). Contusion The increase of GAS and decrease of SS may play an important role in the occurrence and development of DGP.

Objective To compare the quality of life before ischemia postconditioning and 6 months after ischemia postconditioning in patients with acute ST segment elevation myocardial infarction,and investigate the related risk factors.Methods One hundred and twenty-nine patients with acute ST segment elevation myocardial infarction were given the ischemie postconditioning.The quality of life was evaluated before ischemia postconditioning and 6 months after ischemia postconditioning by 36-item short-form health survey (SF-36) and seattle angina questionnaire (SAQ).Results In acute ST segment elevation myocardial infarction patients treated with ischemia postconditioning,the scores of SF-36 and SAQ were significantly lower at 6 months after ischemia postconditioning than those before ischemia postconditioning [SF-36:total score of body health (76.4 ± 17.3) scores vs.(56.3 ± 16.8) scores,physiologic function (75.3 ± 18.1) scores vs.(52.4 ± 19.2) scores,physiologic functional authority (75.6 ± 16.5) scores vs.(48.5 ± 20.3) scores,body pain (77.2 ± 15.4) scores vs.(58.7 ± 16.6) scores,total health state (73.6 ± 17.8) scores vs.(50.6 ± 14.7)scores,total score of mental status (77.5 ± 15.3) scores vs.(55.3 ± 17.3) scores,vitality (69.3 ± 18.1)scores vs.(43.2 ± 17.4) scores,society function (70.3 ± 17.5) scores vs.(41.3 ± 14.2)scores,affection functional authority (80.2 ± 17.4)scores vs.(63.5 ± 14.3)scores,mental health (77.6 ± 18.4) scores vs.(55.6 ± 20.1) scores,health change (76.3 ± 17.4) scores vs.(53.4 ± 16.6) scores;SAQ:body limitation of activity (78.46 ± 12.21) scores vs.(35.34 ± 15.33)scores,angina stable state (74.23 ± 8.53) scores vs.(30.12 ± 5.38) scores,angina episode state (72.34 ± 10.33) scores vs.(27.33 ± 9.12) scores,satisfaction degree of treatment (76.42 ± 12.13)scores vs.(30.56 ± 15.57) scores,knowledge of diseases (74.22 ± 9.35)scores vs.(37.25 ± 20.32) scores] (P ＜ 0.05).The results of multiple linear regression equation showed that the older,more severe vascular lesions,and lower total score of body health score before ischemia postconditioning indicated lower total score of body health,and worse quality of life after ischemia postconditioning.Conclusions The quality of life of patients with acute ST segment elevation myocardial infarction after ischemia postconditioning is significantly improved,compared with that before ischemia postconditioning.Advanced age,vascular disease and lower total scores of body health before ischemia postconditioning are independent risk factors of total scores of body health after ischemia postconditioning.

Aim To set up a novel animal model of intra-abdominal adhesion and to determine whether the tissue plasminogen activator activity (PAA) in exudate can be taken as an indicator to judge the formation of the adhesion. Methods　Rats were randomly divided into 2 groups. Each animal in both groups was opened the abdominal cavity via midline laparotomy without any disinfectant measures. 2-cm section from the cecal end was clamped and ligated, 1-cm cecum of the section was cut, and another 1-cm end from the ligated site was kept. After the content in the end was extruded, the cecum was put back without using any antibacterial agent. Before the skin closure, an effective combination AMD (allantoin, metronidazole and dexamethasone in combination), was given (ip) according to 1.5 ml per 100 g body weight (60.6 mg·kg-1). The control group was injected (ip) the same volume of normal saline. After 6 h, the exudate was extracted from drainage-tube, with the rats varying posture, and after 1 kw, the rats were killed for examining the intra-abdominal adhesion. The values of PAA of exudate in both groups were analyzed by the relative operating characteristic curve (ROC). Results　In the control group, all 20 rats formed the adhesions, the amount of exudate=(1.25±0.09) ml, the number of WBC(×103)=(23.1±6.6) mm3 and PAA=(0.9±0.4) IU·ml-1 in the exudate of abdominal cavity. In AMD group, however, there was not the adhesion formations (0/20), the amount of exuade was (0.52±0.04) ml (P<0.01), the number of WBC (×103) (10.6±4.2) mm3 (P<0.01), and PAA (23.1±6.6) IU·ml-1(P<0.01) in exuade.ROC analysis indicated that if PAA >1.24 IU·ml-1 in the exuade, the adhesion was difficult to form, and vice versa. Conclusion　This animal model can be taken as an effective tool to evaluate the human adhesion related to multi-links on the pathogenesis, and the PAA in exudate an indicator to judge intra-abdominal adhesion formation.

Objective To investigate the clinical characteristics between elderly and non-elderly patients with ulcerative colitis (UC) in order to provide the base for diagnosis and management of the disease.Methods Two hundred and four patients with UC admitted to Shengjing Hospital from January 2009 to December 2011 were enrolled in this study.The patients were divided into elderly group (≥60 years of age) and non-elderly group(＜ 60 years of age).The clinical manifestation,laboratory examination indices and endoscopic feature were recorded and analyzed.Results Among the 204 cases with UC,33 (16.2％,33/204) were elderly patients,and 171 (83.8％,171/204) were non-elderly patients.Onset peak of U C was ranged from 30-39 years old.There was no significant difference in terms of other indices of clinical presentations,extraintestinal manifestations,complications,clinical and endoscopic score between two groups.The rate of abdominal distension,constipation,difficult defecation,alteration of intestinal flora,hypokalemia and hypoproteinemia in elderly group were 36.4％,9.1％,6.1％,66.7％,45.5％,36.4％ respectively,higher than that in in nonelderly group (14.6％,1.2％,0.6％,48.0％,14.0％,20.5％ respectively),and there was significant differences between groups (x2 =8.808,7.260,5.724,3.876,17.660 and 3.942 respectively,P ＜ 0.05).Endoscopic examination revealed that the colonic lesion parts in elderly patients was limited in left-side colon and protosigmoid,which was different from that of no-elderly group (P =0.044).Conclusion The elderly patients showed a similar clinical manifestation with non-elderly patients.However,the colonic lesion parts in elder group was limited to left-side colon and protosigmoid.They also had the higher rate of hypoproteinemia,hypokalemia and alteration of intestinal flora,which needed to pay more attention.

Antibodies against LSP in the sera of 168 patients with various types of viral hepatitis were determined with SPA-RIA. The sera of another group of 178 patients (109 of them were from the first group) with viral hepatitis were studied with ELISA for the same antibodies, which were further divided into three categories, that is, IgG, IgM and IgA classes. The results of 109 patients examined with both of the two methods, indicated that anti-LSP antibodies measured by SPA-RIA might mainly represent anti-LSP IgG class. It was found that circu-lating anti-LSP antibodies could easily be detected in most patients with either acute or chronic hepatitis. After analyzing the-results, the authors suggest that the humoral immune response against LSP might not be the sole initiating factor in the pathogenesis of viral hepatitis, they are more likely the result of the antigen variation of the injured liver cells.

AIM: To investigate the therapeutic effect of tea polyphenols (TP) on cationic bovine serum albumin (C BSA) glomerulonephritis. METHODS: C BSA glomerulonephritic model was induced in rabbits. TP in three different doses (30, 100 and 300 mg?kg -1 ) was administered (ig) once daily for 14 days. RESULTS: TP not only significantly reduced the urinary protein excretion, decreased blood urea nitrogen (BUN) and plasma creatinine (Cr) levels, but also significantly relieved gloermular lesions in the rabbits treated and there was a significant dose dependent relationship between high dosage (300 mg?kg -1 ) and low dosage ( 30 mg?kg -1 ). CONCLUSION: TP can reduce proteinuria, suppress the development of glomerular impairments, and ameliorate the kidney function of rabbits with C BSA glomerulonephritis.

Objective To investigate the effects of environmental enrichment (EE) on the neurological functions in patients with stroke. Methods From May, 2013 to November, 2015, 76 patients with stroke were selected and randomly divided into control group (n=39) and EE group (n=37). Both groups received routine therapy and the EE group received EE in addition. They were assessed with simplified Fugl-Meyer Assessment (FMA), Montreal Cognitive Assessment (MoCA) and modified Barthel Index (MBI) before and after treatment. Re-sults The scores of FMA, MoCA and MBI increased in both groups after treatment (t>2.339, P<0.05), and the scores of MoCA and MBI in-creased more in the EE group than in the control group (t>2.187, P<0.05). Conclusion EE could promote the recovery of cognitive function and activities of daily living in patients with stroke.

Objective To investigate the expression level of collagen triple helix repeat containing 1 ( CTHRC1) in human gastric carcinoma and the relationship with the clinicopathological characteristics of gastric cancer?Methods The expression of CTHRC1 in human gastric carcinoma and normal gastric mucosa were detected by immunohistochemistry ( S?P method ) , and the correlation with various clinical characteristics, including gender,age,tumor diameter,degree of differentiation,depth of invasion,lymph node metastasis,TNM stage,was analyzed?Results ( 1 ) CTHRC1 expressed positive for 41 cases ( positive rate=53?95%) in 76 gastric carcinoma specimens, but only 1 case ( positive rate=3?33%) expressed positive in 30 normal gastric mucosa,the difference was statistically significant (χ2 =23?0332, P=0?000 )? ( 2 ) In early stage of gastric carcinoma,CTHRC1 was predominantly positive in the nucleus,but with the progression of the tumor,CTHRC1 expressed predominantly in cytoplasm?( 3) The expression of CTHRC1 was correlated with the depth of invasion (P=0?000),lymph node metastasis(P=0?009) and TNM?stage(P=0?007),but not with age,gender,size of the tumor and differentiated degree ( P>0?05 )?Conclusion CTHRC1 might play important roles in the occurrence,invasion and metastasis in human gastric carcinoma,and may be new therapy targets.

BACKGROUND: Pathophysiological mechanism of local microenvironment is complex after central nerve injury; especially,both inflammatory reaction at an acute phase and formation of secondary glial scar have tremendous effects on effective regeneration of axon, regeneration and arrangement of local nerve cells, proliferation and migration of local stem cells;therefore, it becomes a basic reason for blocking nerve repair in an early period. Thus, how to effectively resist inflammatory factors in injured region at an acute phase and how to optimize microenvironment of neural regeneration are the most important strategies for repairing spinal cord injury in recent years.OBJECTIVE: To design, establish and screen the best expression of interleukin-6 receptor (IL-6R) α to inhibit shRNA adenovirus expression vector by using spinal cord injury models.DESIGN: Duplicative measurement study.SETTING: Department of Spine Surgery, the Second People's Hospital of Shenzhen.MATERIALS: A total of 40 healthy Wistar rats, either gender, 8-10 weeks old, were selected in this study. Rabbit-anti-rat glial fibrillary acidic protein (GFAP) antibody Ⅰ was provided by Santa Cruz Compan; siRNA eukaryon expression plasmid pGenesil (cohtaining green fluorescent expression system) was provided by Wuhan Jingsai Bioengineering Company.METHODS: The experiment was carried out in ImmuneOpening Laboratory, Basic Medical Faulty, Tongji Medical College, Huazhong University of Science and Technology, and Medica Laboratory Center, the Second People's Hospital of Shenzhen in November 2006. Three pairs of shRNA template which composed of 19 bp reverse repeated motif of IL-6 receptor (IL-6R) α target sequence with 9 bp spacer were designed and synthesized, then the recombinant adenovirus expression vectors with green fluorescence protein were constructed in vitro respectively. The acute spinal cord injury models were completed, and the adenovirus recombinants were regionally injected post 12 hours after spinal cord injury;in addition, the inhibitory effect of RNA interference (RNAi) on expression of IL-6R in local region after spinal cord injury were detected by using real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot so as to screen adenovirus expression vector which had the best inhibitory effect on expression of IL-6R.MAIN OUTCOME MEASURES: Inhibitory effect of RNAi on expressions of IL-6R RNA and protein in local region after spinal cord injury.RESULTS: Sequence analysis showed that IL-6R-shRNA recombinant adenovirus expression vector was successfully constructed, and optimal IL-6R-shRNA recombinant adenovirus expression vector was screened by using real-time fluorescence quantitative RT-PCR and Western blot. The IL-6R expressions were 49% and 56% at the levels of mRNA and protein, respectively.CONCLUSION: The IL-6R--shRNA recombinant adenovirus expression vector is successfully constructed and screened.The gene expression of IL-6R can be highly inhibited after acute spinal cord injury.