Sorting nexins (SNXs) are a large group of proteins that contain Phox (PX) domain and involve in regulating endocytosis and endosome sorting. SNX7, a member of SNXs family, contains a PX domain and a BAR domain. In zebrafish, SNX7 is a liver-enriched anti-apoptotic protein and indispensible for the liver development. A fragment of SNX7 cDNA ((px-bar)snx7), encoding the PX domain and the BAR domain, was inserted into the expressing vector p28a, transformed into E. coli Rosseta 2 (DE3), and then induced by isopropyl β-D-1-Thiogalactopyranoside (IPTG). After affinity, ion exchange and gel filtration purification, the purity of (PX-BAR)SNX7 reached over 95%. Dynamic light scattering (DLS) experiment indicated that (PX-BAR)SNX7 was homogeneous in solution. Lipid overlay assay showed that (PX-BAR)SNX7 can bind to PtdIns(5)P, PtdIns(4,5)P2 and PtdIns(3,4,5)P3.
Escherichia coli ; metabolism ; Genetic Vectors ; Humans ; Phosphatidylinositols ; metabolism ; Recombinant Proteins ; biosynthesis ; Sorting Nexins ; biosynthesis ; Substrate Specificity

Objective To investigate the effect of somatic sense interactive game Kinect on executive dysfunction for patients with stroke. Methods 78 inpatients with first stroke were divided into intervention group and control group, and 35 cases in the intervention group and 36 cases in the control group finished the study. The control group accepted conventional rehabilitation, and the intervention group played the game of Sailing for Gold Coins and Golf of the Kinect in addition. They were assessed with Behavioral Assessment of Dysexecutive Syndrome (BADS) before and 4 weeks after intervention. Results The difference of scores of all the sub-scales and total of BADS before and after intervention were more in the intervention group than in the control group (P<0.05). Conclusion The somatic sense interactive game Kinect can be used to improve the executive function of the patients with stroke.

Objective:To analyze the consistency of HER-2 expression among endoscopic biopsy and radical operation specimens of gastric adenocarcinoma and to investigate the clinical application value of HER-2 detection in trastuzumab treated patients. Methods:From March 2013 to February 2014, 167 patients from Shanghai Changhai Hospital were diagnosed with gastric adenocarcinoma using endoscopic biopsy specimens. The corresponding surgical specimens were collected for pathological analysis. The relevant clinical and pathological data were collected. HER-2 protein expression of endoscopic biopsy specimens was detected by immunohistochemistry (IHC). HER-2 protein expression and gene amplification status of the corresponding tumor resection specimens were detected by IHC and fluorescence in situ hybridization (FISH). Results were analyzed for clinicopathological characteristics. Results:Among the 167 cas-es, 18 cases (10.8%) were HER-2 positive, including 10 cases showing IHC3+and 8 cases showing IHC2+with FISH positive. The consis-tency rate result among endoscopic biopsy and surgical operation specimens was 82%. Excluding the cases showing IHC2+, the true positive rate and the true negative rate were 73.3%and 97.0%, respectively. Conclusion:HER-2 detection of endoscopic biopsy speci-men by IHC shows great predictive value. The main reason for the difference of surgery and biopsy specimens is the heterogeneity of tumor expression. Increasing the number of specimens and combined testing with FISH are important methods to reduce misjudge-ment.

Objective To investigate the effect and mechanism of STAT3 gene silencing on the growth of xenografts in human pancreatic cancer SW1990 cells in nude mice.Methods The expression vector inserted with shRNA targeting at STAT3 gene was constructed and was stably transfected into SW1990 cells (SW1990-RNAi group).SW1990 cells transfected with negative control shRNA expression vector (SW1990-Con group) and parent SW1990 (SW1990 group) were used as controls.STAT3,VEGF,MMP-2 protein expressions in these groups were determined by using Western blot.The subcutaneous xenografts models were established in nude mice,and the growth of xenografts was observed,CD34 expressions were determined by immunohistochemistry and MVD was measured.Results The expression of STAT3 protein was 84.69 ± 9.31,82.00 ± 7.76,7.93 ± 1.24,repectively,in SW1990 group,SW1990-Con group,SW1990-RNAi group,and the expression of VEGF protein was 82.94 ± 8.97,80.86 ± 10.28,39.04 ± 6.23,respectively,and the expression of MMP-2 protein was 40.88 ± 5.09,38.26 ± 5.71,12.54 ± 2.15,respectively.The expression in SW1990-RNAi group was significantly lower than those in other 2 groups (P ＜ 0.05),while the expression of all three proteins between SW1990-Con group and SW1990 group was not significantly different.The weight of the xenografts in SW1990 group,SW1990-Con group,SW1990-RNAi group was (2.2 ± 0.4),(2.2 ± 0.3),(0.5 ± 0.3) g,respectively ; the MVD of the xenografts was (20.35 ± 2.41),(18.79 ± 1.94),(9.62 ± 1.06) per high power field,respectively,and the number in SW1990-RNAi group was significantly lower than those in other 2 groups (P ＜ 0.05 or P ＜ 0.01),while the difference between SW1990-Con group and SW1990 group were not significant.Conclusions Inhibition of STAT3 gene expression can significantly slow the growth of SW1990 xenografts in nude mice,and the mechanism may be related with down-regulation of VEGF and MMP-2 expression and inhibition of the angiogenesis of pancreatic cancer.

To explore the anti-angiogenic effects of genistein on synovium in a rat model of type II collagen-induced arthritis (CIA).

Objective To investigate the effect of genistein (gen) on the secretion of vascular endo-thelial growth factor (VEGF) and matrix metalloproteinases (MMPs-1, 2, 3, 9) by fibroblast-like synoviocytes (FLS) from rats with collagen Ⅱ induced arthritis (CIA). Methods The CIA was induced by collagen Ⅱ and complete Freund's adjuvant (CFA) into rats. The rats were scored based on the arthritis index(AI) once a week. At the sixth week, the X-ray of joints was taken. The synovial tissues from knee joints were examined pathologically. The primary fibroblast-like synowocytes were separated from the synovial tissue by collagenase digestion and cultured. Then the expression of VCAM-1 was estimated by flow cytometry. After adding gen (100, 200, 400 μmol/L) at different concentrations into the FLS, VEGF and MMP-1, 2, 3, and 9 of the supernatants were tested by indirect ELISA. Results After 3 days of type Ⅱ collagen and CFA injec-tion, the rats started to catch arthrocele and their arthritis index increased gradually. The arthrocele was most remarkable at the 3rd week. The AI, X-ray and pathological examination indicated that the model group were significantly different from the control group. After the synoviocytes were cultured to the 4th generation, the expression of VCAM-1 was as high as about 85.5%. It showed that most synoviocytes were changed to fibro-blast-like synoviocytes. Different concentrations of gen (100, 200, 400 μmol/L) added to FLS were compared and revealed that the VEGF and MMP-1, 2, 3, and 9 in the supematants were suppressed evidently and in a dose-dependent manner. Conclusion The CIA model can be successfully constructed by collagen type Ⅱ and CFA. Tthe primary FLS of rats' joint can be separated and cultured well by collagenase digestion. Certain levels of gen can suppress the secretion of VEGF and MMP-1, 2, 3 and 9 hy FLS. The affect is dose-depen-dent.

Objective To observe the formation of the biofilm in endotracheal tubes,the characteris-tics of etiology, drug resistance and relationship between the biofilm and ventilator-associated pneumonia ( VAP) . Methods A total of 60 cases of ventilated children patients whose mechanical ventilation time were≥48 h in the ICU from September 2010 to September 2012,according to the mechanical ventilation time,all cases were divided into 2 to 6 d group,7 to 14 d group and ≥15 d group. The incidence of VAP, biofilm structure under the electron microscope,etiology culture positive rate of the lower airway secretions and bio-film,etiological characteristics and drug resistance were prospectively studied. Results ( 1 ) A total of 19 cases occurred VAP in 60 cases of mechanical ventilation,the incidence of VAP was 31. 7%. (2) Observed by electron microscope,biofilm had formed in the endotracheal tube inner wall in early period of mechanical ventilation. With prolonged mechanical ventilation,biofilm structure had improved,as well as VAP incidence rate from 9. 1%(2 to 6 d group ) increased to 44. 4%(7 to 14 d group) and 88. 9%(≥15 d group). (3) A large number of pathogenic bacteria colonized in the biofilm. Gram-negative bacilli were dominate and drug resistance was high. (4) With prolonged mechanical ventilation,the cultured pathogens from the lower airway secretions and biofilm converged. Conclusion Biofilm could form in the endotracheal tube with mechanical ventilation patients,and is associated with the occurrence of VAP and refractory infections.

matK is one of the core DNA barcode markers for plant DNA barcode identification and its universality using single makers has been in controversy. However, the universalities of different matK primer pairs in same seed plant group (order) and same matK primer pairs in different seed plant groups (order) are lack of systematic research. In this study, we collected 14563 full-length matK sequences of 11429 species of 3292 genera in 239 families belonging to 36 orders in seed plants. The universalities of 13 matK primer pairs and its 78 primer com-binations have been assessed using bioinformatics methods. The results indicated that xf/5r, 1F/8R, 390F/1326R and 3F_KIM/1R_KIM were the four most universality primer pairs. The four markers' universalities were 91.18%, 84.65%, 79.81% and 80.94% respectively in all 11429 seed plants. The most universality primer pairs in different orders were different. For each order, the primer pair with maximum universality was different. the xf/5r was the basal primer pair for primer combination and 1F/8R, 1F/1R, M3/M4 and 3F_KIM/1R_KIM could be the complementary primer pairs. This study could be a valuable resource for the primer selection of the research DNA barcoding identification in seed plants.

Aim To study the anti-stress depression effect of Jiaweisinisan(JWSNS)and it's mechanism of N-methyl-D-aspartate(NMDA)receptor channel in hippocampus.Methods Chronic mild unpredictable stress(CMUS)was adopted to establish the rat depres-sion model.Before and after model establishing,the sucrose consumptions and the weight of rats were detected.TTC dyeing of hippocampus brain slices ex vivo were used to detect the situation of hippocampus neuron damage and the protective effect of serum containing components of JWSNS.The opening probability and the average opening time of NMDA receptor channels were detected by attach on cell patch clamp,the Ca2+ concentrations in single cell of hippocampus neurons were detected by Tillvis ION software.Results CMUS could decrease the sucrose preference and the weights of rats(P

Objective: To explore the relationship between atrial MMP-9 with its inhibitor (TIMP-1), anti-apoptosis gene (BCL-2) with apoptosis gene (BAX) and the aging with atrial remodeling in experimental dog model during atrial ifbrillation (AF), in order to better deal with the aging caused AF.
Methods: The experimental dogs were divided into 4 groups: ①Adult with sinus rhythm (ASR) group, ②Elder with sinus rhythm (ESR) group and③Adult with AF (AAF) group,④Elder with AF (EAF) group. n=7 in each group. Chronic AF model was induced by rapid and persistent atrial pacing. The mRNA and protein expressions of MMP-9, TIMP-1 and BCL-2, BAX were measured by real time quantitative RT-PCR and Western blot analysis. The cellular ultra structural remodeling was examined by optical/electron microscopy, and the apoptosis index was determined by TUNEL method,
Results: Compared with adult dogs, the elder dogs showed obviously increased expressions of MMP-9, BAX, and decreased expressions of TIMP-1, BCL-2, all P<0.05. Compared with SR gods, the AF dogs presented up-regulated expressions
of MMP-9, BAX, all P<0.05, and down-regulated expressions of TIMP-1, BCL-2, all P<0.05, such changes were most obvious in elder AF dogs. Accompanying with the aging and AF, the degree of atrial ifbrosis, cellular ultra structure and the apoptosis index were changed with the statistic meaning.
Conclusion: The abnormal expressions of MMP-9/TIMP-1 and BCL-2/BAX might be one of the molecular mechanisms for aging caused AF in experimental dog model.