Objective To investigate the pathogenesis of portal hypertensive gastropathy(PHG). Methods Two rat models with portal hypertension(PHT) and a sham operation group were established to detect the pathological changes in histology and ultrastructure of the gastric mucosa as well as quantitative changes in histological morphology by graphic analysis computer system. Results Prominent edema, scattered red dots/ecchymosis were found in gastric mucosa in rats with PHT. Light and scan electron microscopy showed swelling or exfoliating of the epithelium cell, reduction of gastric gland number, thin of gastric mucosa, while infiltration of inflammatory cells, epithelium metaplasia were not found. The most characteristic findings were the changes of the mucosal capillaries and venules in both mucosal basal lamina and submucosa layer light microscopically, as well as the transmutation, stenosis and loose intercellular joining of the capillaries on electron microscopy. Ultrastructure observation revealed numerous pinocytes in epithelial cells as well as proliferation and hyperplasia of smooth muscle, collagenous fiber and extracellular matrix in venules. Quantitative analysis showed that the ratios of the cross sectional area and the vascular wall area between the gastric wall area(CSA/GWA & VWA/GWA) was higher in liver cirrhosis(LC) and portal vein stenosis(PVS) groups than that in sham operation(SO) group. There was a positive correlation between portal vein pressure and the ratio of the length of damaged mucosa and the length of mucosa(LDM/LM that was higher in LC group than in PVS group). Conclusions In rats with cirrhotic portal hypertension, distinct gastric microvascular morphological changes are the major etiological factor of PHG as a part of pathological changes in portal hypertension.

OBJECTIVETo study the proliferation and differentiation of neural stem cells in the subventricular zone (SVZ) in neonatal rats after bilateral common arteries occlusion.

METHODSNinety-six 3-day-old Sparuge-Dawley rats were randomly divided into two groups: ischemia and control. Rats in the ischemia group were subjected to bilateral common arteries occlusion and the rats in the control group were sham-operated. All rats were administrated with 5-bromodeoxyuridine (BrdU) (50 mg/kg) via intraperitoneal injection. Rats were sacrificed and their brains were removed 1, 4, 7, 10, 14 and 35 days after ischemia. Using brain paraffin sections and immunofluorescence assays, the number of newborn cells in the SVZ was counted. Newborn neural stem cells and oligodendrocytes in the SVZ were observed, and then double marked with BrdU and nestin or osmium tetroxide (O4).

RESULTSThe number of BrdU+ cells (neural stem cells) in the SVZ in the ischemia group was greater than in the control group 4, 7, 10 and 14 days after ischemia, and reached a peak at 4 days after ischemia (253.1+/- 49.3 vs 133.5+/- 17.7; P< 0.01). By 35 days after ischemia, the number of BrdU+/O4+ cells (oligodendrocytes) in the corpus callosum (56.0+/- 7.2 vs 17.0+/- 6.4; P< 0.01), the septal nuclei (45.0+/- 11.9 vs 20.5+/- 5.0; P< 0.01), the striatum (34.5+/- 4.2 vs 14.5+/- 5.8; P< 0.01) and the olfactory bulb (46.5+/- 6.6 vs 23.5+/- 8.4; P< 0.01) in the ischemia group increased significantly as compared to the control group (P< 0.01).

CONCLUSIONSBrain ischemia can activate the proliferation of neural stem cells in the SVZ and promote neural stem cells differentiation into oligodendrocytes. The immature brain may have the capacity for self-repair after ischemic brain injury.

Animals ; Animals, Newborn ; Brain Ischemia ; physiopathology ; therapy ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Female ; Male ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation

OBJECTIVE Dynamics of serum and urine metabolites in hepatic injury rats induced by ethanolic extract from Rhizoma Dioscoreae Bulbiferae(RDB)was investigated by 1H-NMR-based metabo?nomic methods in order to discover early biomarkers of liver toxicity induced by RDB. METHODS Rats were ig adminisetred with RDB at a dose of 5 g·kg-1 for 28 d. Rats were sacrificed 3,7,14 and 28 d af?ter RDB administration,as well as after a recovery period,respectively. Blood was taken for routine bio?chemical analysis by an automatic biochemical analyzer. Liver/body mass indexes were calculated ,and liver pathological changes were observed with hematoxylin-eosin staining. Urine samples were collected before and 3,7,14 and 28 d after RDB administration,respectively,as well as after withdrawal. Metabo?nomic analysis was carried out for serum and urine samples. Principal component analysis and orthogonal partial least squares-discriminant analysis were used for screening and identifiying early biomarkers. RESULTS Compared with the control group,total bilirubin (TB) and total cholesterol (TC) values were increased in 3-28 d in RDB group(P<0.05,P<0.01). Total bile acid(TBA)was elevated in 7-28 d (P<0.05,P<0.01). TB,TC and TBA became normal after discontinuation with RDB. There was no significant difference between RBD-treated group and control group in the activity of glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase,and the content of glucose also was not different between the two groups. The ratio of liver/body mass was elevated at 3-28 d(P<0.01)but returned to normal after withdraval of RDB. The enlargement and necrosis of hepatocytes were observed 7 d after RDB administration,and lesion degree was aggravated with the extension of RDB delivery time. Meta?bonomic analysis showed that the serum lipids (low density lipoprotein/very low density lipoprotein (LDL/VLDL),glutamic acid,choline phosphate and glycerolphosphatecholine were increased in the early stage. Pyruvate and N-acetylglutamate were decreased in urine. These metabolites became normal 7 d after discontinuation with RDB. CONCLUSION The serum lipids (LDL/VLDL),glutamic acid,glycerol phosphate choline,as well as urine pyruvic acid salt and N-acetyl glutamate may be used as the early biomarkers for liver toxicity induced by RDB.

Objective To establish a key technological system for spider fibroin gene code tandem connection , vector construction , prokaryotic expression and purification using genetic engineering in order to achieve MaSp 1 heterologous ex-pression in Escherichia coli and its separation and purification .Methods Isocaudarner ligation method was used to connect synthetic spider fibroin gene monomer code in tandem , and a recombinant clone concatemer was obtained .The identified recombinant clones were connected with prokaryotic expression vector pET 28a(+), and then transformed into E.coli BL21 (DE3).After being induced by IPTG for 6 hours, the expression product was identified by SDS-PAGE and Western blot-ting.Engineering bacteria were fermented in high density , and the obtained protein was purified through ammonium sulfate fractionation.Results and Conclusion The expression plasmids of MaSp1concatemers were successfully constructed , and the induced expression genetic engineering MaSp 1 protein was of the expected relative molecular mass .In addition, the pu-rity of the purified protein was above 80%.This study has developed crucial technologies for mass production of genetic en-gineering spider silk proteins .

Objective:To explore the effect of SimMan 3G simulator based scenario simulation teaching method and case-based learning (CBL) in emergency medicine teaching.Methods:Sixty students from Batch 2013 eight-year program of clinical medicine were selected as subjects. They were randomly divided into an experimental group and a control group, with 30 students in each group. In the teaching of emergency medicine, the experimental group used the combination of scenario simulation with CBL teaching methods, and the control group used classic teaching methods. The test scores and the questionnaires satisfaction of the two groups were compared to evaluate the teaching effects. SPSS 17.0 was used for the statistical analysis, measurement data were compared between the groups by t test, and counting data were compared between groups by chi-square test.Results:The scores of the experimental group (94.24±1.13) were better than those of the control group (90.6±0.59), with significant differences ( t=12.85, P<0.05). The results of the questionnaires showed that the students of experimental group were more satisfied with the learning experience than those of the control group. Conclusion:The teaching method can improve the teaching effects, the students' emergency clinical thinking, skills, comprehensive analysis and judgment ability, team cooperation consciousness and leadership ability.

Objective To investigate the effect of amiloride on the invasion capacity of esophageal carcinoma EC9706 cell line in vitro and to elucidate its possible mechanism.Methods The invasion capacities of EC9706 cells pretreated with amiloride were measured by transwell chamber assay. The urokinase-type plasminogen activator (uPA) transcription were determined by RT-PCR.The protein expression of uPA were assessed by Western blot.Results After the EC9706 cells were pretreated with amiloride at different concentrations,the number of invaded cells was obviously less than those of control group with obvious dosage dependent pattern (96±7,78±6,57±6,33±4,15±3,F =43.46,P ＜ 0.01).The transcription levels of uPA mRNA and the protein expression levels of uPA in EC9706 cells decreased significantly compared with the control (mRNA:0.623±0.065,0.526±0.054,0.389±0.041,0.312±0.038,0.247±0.025,F =6.71,P ＜0.01; protein:0.732±0.064,0.644±0.057,0.533±0.058,0.391±0.036,0.267±0.043,F =6.71,P ＜0.01).Conclusion Amiloride inhibits the invasion capacity of esophageal carcinoma EC9706 cells.The mechanism might be associated with down-regulation of the expression of uPA.

Objective To investigate the expression and influence to tumor angiogenesis of urokinase-type plasminogen activator (uPA) and vascular endothelial growth factor (VEGF) in esophageal carcinoma. Methods The expression of uPA and VEGF in the tissue of normal (18 cases) and esophageal carcinoma (68 cases) were evaluated by SP immunohistochemistry, CD34 was detected as marking tumor microvessel density (MVD). uPA and VEGF expression were assessed as to the pathologically biological features of esophageal cancer and to the influence to tumor angiogenesis. Results The positive rates of uPA were 27.8 % (5/18) and 70.6 % (48/68) in the tissue of normal and esophageal carcinoma, respectively, there was significant difference in two tissues (x2 =11.63, P <0.05). The positive rates of VEGF were 22.2 % (4/18)and 63.2 % (43/68) in the tissue of normal and esophageal carcinoma, respectively, there was significant difference in two eissues (x2 =9.78, P <0.05). The expressions of uPA and VEGF in esophageal carcinoma were uniformity (x2 =9.72, P <0.05). The mean of MVD was 42.38±11.62. The positive rates of uPA and VEGF were higher in the high MVD group than those in the low MVD group (x2 =6.13, P <0.05, x2 =10.12, P <0.05,respectively). uPA and VEGF expressions in malignant tumors weren' t associated with age, gender and pathological types (P >0.05), but associated with clinical stage, histologic grading and lymph node metastasis (P <0.05). Conclusion Rising expression levels of uPA and VEGF are common in esophageal carcinoma. Altered expression of uPA and VEGF may contribute to tumor angiogenesis of esophageal carcinoma, whose overexpression indicate worse prognosis.

Objective:To amendment a tool of theory of mind (TOM) tests that can be applied to children with autism spectrum disorder (ASD) in mainland China,and assess the ability of ToM of children with ASD.Methods:The items of the ToM tests were revised by observing and recording events of children's life.Totally 200 normal children were selected in line with the standard from kindergarten to Grade 6 for formal tes-ting.With 156 valid data,Pearson correlation coefficient and Cronbach α coefficient were established for the test.Three experts were invited to make sure the content validity.Researchers randomly selected 30 normal children for retest purpose after 2 months.Twenty five children with ASD were tested and compared with normal children's test scores.Results:The revised test included 39 entries,which was divided into three sub-tests,42 points in total.The correlation coefficient of three subtests of the tests was 0.54-0.77,the correlation coefficient between the test and the subtest was 0.62-0.93 (Ps ＜ 0.01).The scores of three experts for the test were 114,108,and 105.The total scores and subtest scores were lower in children with ASD than in normal children (Ps ＜0.01).The Cronbach a coefficient of the test was 0.84,Cronbach a coefficients for three subtest were 0.83,0.80,and 0.78,respectively.The retest reliability was 0.84,and reliabilities for three subtest were 0.75,0.74,and 1.00.Conclusion:The revised Theory of Mind Tests for Children with Autism Spectrum Disorder are fulfilled mostly in line with psychometric testing requirement.It might be a selection to measure the ability of theory of mind of children with ASD in mainland China.

Objective:To investigate the relationship between Creactive protein(CRP)/albumin ratio (CAR) and lymph node metastasis of gastric cancer. Methods:A total of 96 cases of gastric cancer were included in the study. The clinical pathological stage and lymph node detection in patients with gastric cancer were analyzed, with the preoperative CAR as the dependent variable. Results:1) The pa-tients with preoperative CAR>0.04 has higher transfer rate and metastasis than the patients with preoperative CAR≤0.04, and the dif-ference was significant (P<0.05). The lymph node stage of patients with preoperative CAR>0.04 was significantly higher than that of the patients with preoperative CAR≤0.04, and the difference was significant (P<0.05). 2) The mean CAR of patients with Borrmann typesⅠ,Ⅱ,Ⅲ, andⅣgradually increased;with the progression of the pathological stage of gastric cancer, the mean value of CAR in-creased. 3) During the operation, the total number of lymph node dissections was high and the mean value of CAR before the opera-tion was high. Conclusion:The correlation between preoperative CAR and lymph node metastasis in patients with gastric cancer may reflect the degree of lymph node metastasis to a certain extent.

Objective To determine the malignant grade related proteins implicated in human astrocytoma with proteomic techniques. Methods Two-dimensional electrophoresis (2-DE) was used to compare the protein expression profiles of normal human brain tissues (n=12) and astrocytoma tissue samples of various malignant grades (23 with grade Ⅱ, 15 with grade Ⅲ and 14 with grade Ⅳ ).After identifying the differential protein spots, electrospray ionization (ESI) tandem mass spectrum and bioinformatics were employed to determine the protein ID. Results A total of 15 proteins with differential expressions were identified,which included 10 up-regulated ones (heat shock protein 27,prohibitin,78-kDa glucose-regulated protein,peroxiredoxin 1,peroxiredoxin 6,annexin Ⅱ,cathepsin D,plasminogen activator inhibitor-1, actin [cytoplasmic 1], Glutathione S-transferase M) and 5 down-regulated proteins (disulfide isomerase A3 protein,alpha-crystallin B chain protein,T-complex protein 1 ε subunit,ubiquitin carboxyl-terminal hydrolase isozyme L1 and glial fibrillary acidic protein) in astrocytoma tissues of grade Ⅲ and Ⅳ. Conclusion Proteomic techniques can screen the malignant-grade associated proteins effectively,which provides rationale for individualized diagnosis and therapeutic targets of astrocytoma.