Objective To explore the structure and type of sugar chain in 33 500 pro-nucleating protein, and its role in gallstone formation. Methods The 33 500 vesicular protein was examined by dot-immunobinding assay of lectin coupled to a peroxidase (HRP-DSA,HRP-ConA,HRP-WGA). The morphology of biliary vesicles was observed under transmission electron microscopy in process of massive vesicular aggregation, culmination and crystal formation. The protein and its enzymatic deglycosylation fractions nucleation promoting activity were detected by cholesterol crystal growth assay. Results 33 500 vesicular protein with multiantennary and complicated glycan displayed apparent potency of nucleation promotion, which clearly reflected by HRP-DSA immunobinding, and derived crystal growth curve indices. It, Ig, Ic were presented as 0.57, 1.52, 1.63 respectively, but after treated by N-glycanase enzyme, no promoting activity was found. Conclusions Our data suggest the sugar chain play an important role in pro-nucleating process, and may be involved in the gallstone formation.

Objective To investigate the effects of angiotensin II type I receptor antagonist losartan on apoptosis and the expression of Fas and Fas-L in the kidney of diabetic rats. Methods Uninephrectomized, STZ(65 mg/kg) -induced diabetic male SD rats were used. Losartan(40 mg/kg) was delivered daily by gavage from the next day of the induction to diabetes for 12 weeks. Apoptosis was examined by means of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL) . Flow cytometry and immunohistochemistry were used to detect the expression of Fas and Fas-L. Results Compared with those in the kidneys of control group, apoptotic cells were more in number and the expression of Fas and Fas-L was higher in the diabetic kidneys( P

OBJECTIVE: To establish a rapid and precise detective method of 33.5 kd vesicular protein and to screen an effective treatment of cholelithiasis. METHODS: Specific antibody of the biliary vesicular protein was obtained by immunizing rabbits and enzyme-linked immunosorbent assay (ELISA) kit was developed. The concentrations of 33.5 kd vesicular protein in serum and bile of gallstone patients and control were examined respectively. The effects of Cholagogue Dry Syrup and Eulektrol Capsule on decreasing 33.5 kd vesicular protein were also studied by ELISA kit. RESULTS: One-step ELISA equation was Y=0.035 X (r=0.99). The vesicular protein concentrations in serum and bile of cholesterol gallstone group [(179.8+/-97.9) mg/L and (213.4+/-70.1) mg/L respectively] were significantly (P<0.05) higher than in the pigment stone group and control. Data showed that, with 2-week administration, Cholagogue Dry Syrup significantly decreased both biliary and serum 33.5 kd vesicular protein of cholesterol gallstone patients, while Eulekrol Capsule and control groups didn't have the same results. CONCLUSION: The concentrations of 33.5 kd protein are different in cholesterol gallstone patients and healthy groups which might be related to cholesterol nucleation process. Cholagogue Dry Syrup is of cholagogic and litholytic effect by decreasing biliary lithogenesis.

The respiratory burst of neutrophils is an important response in inflammation. The alterations in extracellular pH per se is not a sufficient trigger to the respiratory burst in neutrophils. While neutrophils in respose to conventional agonists, the respiratory burst in neutrophils is affected by the alterations in extracellular pH, and there is a direct relationship between intracellular and extracellular pH. O-? 2 production in neutrophils is decreased while H 2O 2 production is increased at acidic pH. O-? 2 release in neutrophils is enhanced at weak alkaline pH. Na+/H+ exchanger plays an important role in the regulation of the respiratory burst of neutrophils.

Objective To investigate the effects of electrical stimulation on the expression of vascular endothelial growth factor(VEGF)mRNA and receptor FLK 1/KDR in a ischemic model of rat hindlimb. Methods The model of hindlimb ischemia on the right side was established by ligation of the superficial femoral artery in 10 rats. The rats were then randomized into an experimental group and a control group. The rats in the experimental group were intervened with electrical stimulation ( 25 Hz, 0.1 V) on the tibialis anterior (TA) of the right side, while those in the control group were not. RT PCR and immunohistological methods were used to detect the expressions of VEGF mRNA and protein in TA muscles. FLK 1/KDR was detected by means of Western blot and immunofluorescence. Results After 7 days of continuous stimulation, there was a significant increase in blood flow within the muscle. VEGF mRNA and VEGF protein had 4 fold and 2 fold increases, respectively, in the stimulated TA muscles as compared to the control(2.58 vs 0.93, 0.48 vs 0.24, P

Objective To investigate the protective effect and mechanism of Astragaloside Ⅳ (AS-Ⅳ) on hypoxia/re-oxygenation induced by RAW264.7 murine macrophages.Methods The hypoxia/re- oxygenation induced RAW264.7 murine macrophages served as a model of I/R injury. The cells were divided into the control group, the model group, and the AS-Ⅳ group. The cell morphological changes of each group were observed directly under inverted microscope. Inducible nitric oxide synthase (iNOS), cluster of differentiation 206 (CD206), and peroxisome proliferator activated receptor-γ (PPAR-γ) were separately examined by RT-PCR, Western blot analysis and immunofluorescence staining.Results Compared with the model group, the numbers of cell in AS-Ⅳ group significantly increased , and the cell physiological status were much better. Compared with the model group, the iNOS immunofluorescence semi quantitative (0.62 ± 0.02 vs. 1.32 ± 0.09), the expression of mRNA (1.51 ± 0.07 vs. 3.46 ± 0.39), and protein (2.30 ± 0.14 vs. 5.16 ± 0.49) significantly reduced in AS-Ⅳ group (P<0.01). The CD206 immunofluorescence semi quantitative (1.01 ± 0.03 vs.0.61 ± 0.01), the expression of mRNA (0.91 ± 0.03 vs.0.51 ± 0.01), and protein (0.61 ± 0.04 vs.0.19 ± 0.01) significantly reduced in AS-Ⅳ group (P<0.01). Compared with the model group, the PPAR-γ immunofluorescence semi quantitative (0.60 ± 0.14 vs. 0.34 ± 0.03), mRNA (2.00 ± 0.14 vs.1.04 ± 0.03), andprotein (0.67 ± 0.05 vs.0.19 ± 0.01) significantly increased (P<0.01). Conclusions The AS-Ⅳ could attenuate ischemia-reperfusion injury by altering macrophages phenotype through upregulation PPAR-γ.

Objective To investigate the relationship between thyroid nodules with calcification and thyroid carcinoma and its significance in the screening of thyroid carcinoma in high risk group.Methods The clinical data of 1771 patient undergoing surgery for thyroid nodules from March, 2006 to March, 2009 in Huashan Hospital, Fudan University were retrospectively analyzed. Results Among 1771 patients, 500 were finally identified as having malignant tumors. Incidence of calcification in thyroid carcinoma was 68. 4%, and that in benign thyroid nodules was 27.0% ( χ2 = 259. 5, P ＜ 0. 05 ). The specificity of microcalcification for the diagnosis of carcinoma was 89. 4%, and its positive predictive value was 66. 3% ( χ2 = 368.6, P ＜ 0. 01 ). The incidence of thyroid carcinoma in patients ＜ 45 years was 39.2%, while that in patients ≥ 45 years was 22.9% ( χ2 = 51.12, P ＜ 0. 05 ). The incidence of carcinoma in patients of single thyroid nodule was 31.7% and that in those with multiple nodules was 26. 4% (χ2 =4. 766,P ＜ 0. 05). Metastasis was pathologically diagnosed in 26. 8% of lymph nodes found by preoperative ultrasonography. Conclusions The specificity of thyroid nodule calcification, especially microcalcification is high for the diagnosis of thyroid carcinoma. High-risk index for carcinoma includes thyroid nodules with microcalcification, ＜ 45 years old and single thyroid nodule.

Objective To evaluate the value of echocardiography in transcatheter closure of ruptured sinus of Valsalva aneurysm(TC-RSVA).Methods TC-RSVA was attempted in 11 patients.The location,shape,size of defects and its relationship with the neighbor structures were revealed before the procedure.Then the deployment of occluder was monitored during the procedure,and the effectiveness was observed in the follow-up.Results Eleven patients were diagnosed as the isolated RSVA by echocardiography.The size of defects was 2 - 13 mm estimated by echo.The procedures were successful in all patients.Usually the Amplatzer duct occluders were chosen to be 1 to 5 mm larger than the size of defects.Three patients had mild residual shunt during the procedure,which all dispeared in the first month of follow-up,but one of them demonstrated recurrent mild residual shunt in the 32nd month of follow-up.There was no aggravating aortic regurgitation in the follow-up.Conclusions TC-RSVA is relatively safe and effective.Observation of long-term effectiveness is still necessary.Echocardiagraphy plays an important role in TC-RSVA.

Objective To investigate the data of gene expression microarray by protein interaction network analysis,establish an interaction network of differentially expressed genes between Kashin-Beck disease (KBD) and osteoarthritis (OA) and choose the central nodes of the network.Methods The articular cartilage samples of degrees Ⅱ ° and Ⅲ ° KBD and OA patients were selected according to the national diagnosis criteria for KBD and the Western Ontario and McMaster Universities (WOMAC) for OA.Chondrocytes of 8 patients with KBD and 7 with OA were selected.About 1 000 different genes detected by gene expression microarray were inputted into STRING 9.1 database online for analysis and establishment of the interaction network.The interaction data were imported into Cytoscape 3.2.1 software for screening the central nodes of the network.KEGG database was exploited for pathway analysis and functional study of the central node genes,Real-time PCR (RT-PCR) was used for verification.Results The protein products of 334 differentially expressed genes between KBD and OA had interrelation,forming a complicated interaction network.About 150 central nodes were selected by Cytoscape 3.2.1 that involved in more than ten signal pathways involved in mitochondria,bone metabolism and inflammatory cytokine.Conclusion The interaction network of the differentially expressed genes between KBD and OA,especially the central nodes of this network,can provide clues to the mechanism and early diagnosis and molecular targeted therapy of KBD and OA.

Objective To evaluate the diagnostic accuracy of ELISA kit for cholesterol gallstone. Methods The ELISA kit of 33?500 vesicular protein was established by sandwich method, and the concentrations of the protein in gallbladder bile were examined among cholesterol, pigmental gallstone patients and control groups. Results The gallbladder 33?500 vesicular protein (213?70) ?g/ml is much higher in cholesterol gallstone patients than in pigmental gallstone patients (72?55) ?g/ml and control groups (65?52) ?g/ml (F=60.9, P