0.05). Of 98 cagE positive isolates, 14(14.3%) were cagA negative. Conclusion Most of the H. pylori isolates in Shanghai region may have intact cag PAI. cagE, could be taken as the marker for the presence of cag PAI. There is no correlation between the integrality of cag PAI and the clinical outcome of H. pylori of infection.

The invasive model of gastric carcinoma in nude mice was constructed with gastric carcinoma cells SGC-7901 and SGC-7901-T2. The expression of type IVcollagenase (MT-MMP) and the changes of basement membrane (BM) in normal kidey tissue and malignant lesion of gastric were studied by immunohistochemical technique. The results showed that the positive rate of type IV collagenase(MT-MMP)was 36.6%in normal kidey tissue and 70% in malignant lesions (P

Objective To investigate the metabolism characteristics and the potetial biomarker candidates of osteonecro?sis of the femoral head (ONFH) using metabolomic technology. Methods The femoral head specimens from 23 ONFH patients (25 necrotic femoral heads) and 18 normal femoral heads from femoral neck fracture patients were collected for histopathological examination to confirm the diagnosis of all samples. All the metabolites of bone trabecula were extracted for ultra-high perfor?mance liquid chromatography-MS/MS analyzed. The measured variables was pretreat, and PCA (principal component analysis), PLS?DA (partial least squares?discriminant analysis) and OPLS?DA (orthogonal?partial least squares?discriminant analysis) models were employed to confirm the difference between these two groups after UPLC?MS/MS (ultra?high performance liquid chromatogra?phy?mass spectrometry/mass spectrometry) analysis. At last, the differential variables were screened out by PLS?DA and variate analysis (Kruskal?Wallis H test). The changed metabolites were confirmed by MS and MS/MS aligned in HMDB (human metabolo?mic database) and Massbank. The changed metabolites with the most obviously changed peak abundance, D?arginine, L?proline and L?glutamine, were picked out as the potential diagnostic biomarkers. After binary logistic regression analysis, the combined biomarkers candidates were further analyzed by receiver operating characteristic (ROC) curve to evaluate the significance of the combined biomarkers. Results Significant distinction of metabolites expression mode can be seen in PCA, PLS?DA and OPLS?DA models scoring plots between ONFH and control groups. Twelve changed metabolites in ONFH bone trabeculas were con?firmed by multi?variate statistical analysis and variate statistical analysis. Compared with the femoral neck fracture patients, the in?creased metabolites included D?arginine, L?proline, L?glutamine, creatine, uracil, uridine, LysoPC(20∶4(5Z, 8Z, 11Z, 14Z)), Ly?soPC(16∶0), PC(20∶1(11Z)/18∶3(6Z, 9Z, 12Z)) and PE(P?16∶0e/0∶0). The decreased metabolites were reticulataxanthin and β?cryptoxanthin. According to the change fold of peak abundance and variable weight projection in PLS?DA, the most obviously dif? ferential metabolites were picked out as the biomarker candidates of ONFH. The potential biomarkers candidates were identified as D?arginine, L?proline and L?glutamine. The area under the curve of D?arginine, L?proline and L?glutamine ROC were 0.873, 0.712 and 0.862. The area under the curve of ROC was 0.946 after combining D?arginine, L?proline, L?glutamine using binary lo?gistic regression analysis. Conclusion PCA, PLS?DA and OPLS?DA models were used to find out the differential variables in the metabolites of bone trabeculas in ONFH and femoral neck fracture patients. Twelve metabolites were identified by MS/MS, and 3 obviously changed metabolites, D?arginine, L?proline, L?glutamine, were indicated as biomarker candidates. These 3 obviously changed metabolites showed a good diagnostic significance.

【Objective】To explore the long-term storage method for the seeds of Aquilaria sinensis(Lour.)Gilg..【Methods】The seeds of Aquilaria sinensis(Lour.)Gilg.with different water contents were preserved with liquid nitrogen for 55 days.And then the seeds were taken out to detect their germination capacity.The effect of cryopreservation methods on the germination capacity was also observed.【Results】The seeds with water content being 16.30% lost their germination capacity completely,the seeds with water content being 13.02% and 9.34% had a lower germination capacity,and the seeds with water content being 7.35% had a higher germination capacity.Quickly-frozen seeds had a higher germination capacity than the slowly-frozen seeds.【Conclusion】Cryopreservation with liquid nitrogen is suitable for the long-term preservation of the seeds of Aquilaria sinensis (Lour.)Gilg.with water content being 7.35%.

Objective To understand awareness of basic knowledge on control and prevention for chronic obstructive pulmonary disease (COPD) among general practitioner in community health service centers. Methods Questionnaire survey was conducted among 593 general practitioners at 15 community health service centers in a district of Shanghai in the morning of February 26, 2008. Results Five hundred and forty-two of 593 ( 91.4% ) general practitioners surveyed knew that chronic obstructive pulmonary disease can be abbreviated as COPD, and 315 of them (53.1%) thought they knew the main contents of the "Guidelines for Prevention and Control of Chronic Obstructive Pulmonary Disease". No pulmonary function instrument was equipped at all 15 community health service centers surveyed, and 501 of 593 ( 84.5% ) general practitioners knew spirometry is gold standard for diagnosis of COPD. Only 115 ( 19.4% ) general practitioners knew that COPD can be categorized as acute exacerbation and stable stage. One hundred and sixty-three of them (27.5%) knew that drug therapy should be maintained in COPD patients for long term following effective treatment, 328 (55.3%) recognized that treatment for COPD patients was still needed in their stable stage, and 312 (52.6%) believed that antibiotics may not be necessary and 465 (78.4%) thought that inhalation therapy was the choice of treatment for acute exacerbation of COPD. Two hundred and thirty-five of them (39.6%) knew that oxygen inhalation should be more than 15 hours per day in regular family therapy. The investigation showed that 365 (61.6%) general practitioners had participated in training for COPD-related knowledge, only one or two times a year. Awareness of COPD knowledge was significantly poorer in rural practitioners than that in urban community ones. Conclusions Basic knowledge of prevention and treatment for COPD was seriously lack in general practitioners, so it is a necessity to strengthen training for them.

To understand molecular modules related to polyunsaturated fatty acids (PUFA) synthesis and eventually produce PUFA at high efficiency, we developed a protein complex analysis technology in Synechocystis sp. PCC 6803, and applied it to identify possible partner proteins interacting with the key enzymes that catalyze PUFA biosynthesis. We first constructed a recombinant expression of protein of slr1609 encoding the fatty acid activation enzyme, by fusing 3xFLAG tag with the target protein. Then we verified its expression by Western blotting targeting 3xFLAG tag. To maximize purification of Slr1609 protein complex, we optimized the protein expression conditions of Slr1609 in Synechocystis in a 5 L fermenter by monitoring its gene expression using RT-qPCR. The purification of the Slr1609 protein complexes was demonstrated by a Native-PAGE analysis. Finally, LC-MS/MS proteomic analysis allowed identification of the possible partner proteins interacting with Slr1609.
Bacterial Proteins ; chemistry ; Chromatography, Liquid ; Fatty Acid Synthases ; chemistry ; Fatty Acids, Unsaturated ; biosynthesis ; Proteome ; chemistry ; Proteomics ; Synechocystis ; enzymology ; Tandem Mass Spectrometry

Objective To introduce surgical repair methods of cross-bridge transplantation of free latissimus dorsi muscular flap and free fibula for complex lower leg defect and discuss its clinical feasibility.Methods The study included 12 patients with tibial defect larger than 8 cm (range,9-12 cm) combined with soft tissue defect of 17 cm × 12 cm to 20 cm × 18 cm treated from May 2008 to May 2012.Cross-bridge transplantation of free latissimus dorsi muscular flap and free fibula was performed at the first phase.The flap pedicled with subscapular vessel was anastomosed to posterior tibial artery and vein of normal lower leg.The flap pedicled with anterior serratus muscle of distal thoracodorsal artery was anastomosed to peroneal vessel of fibular flap.External fixators were used to immobilize the bilateral lower legs postoperatively.Results All patients were followed up for 13-32 months (mean 21 months).According to Enneking system,mean leg function was scored 23 points after tandem transplantation of free latissimus dorsi muscle and free fibula,with recovery rate of 77％.Conclusions Cross-bridge transplantation of free latissimus dorsi muscular flap and free fibula tackles the problem of recipient vessel limitation.Further,the technique is effective in repair of large area of complex defect in lower legs.

This study was aimed to screen candidate genes involved in the triterpenoid saponins biosynthetic pathway of the Ilex asprella root. The Illumina platform was applied to perform transcriptomic sequencing of I. asprella root, followed by a series of bioinformatics analysis. The results showed that a total of 272 candidate unigenes were anno-tated to be involved in the biosynthetic pathway of terpenoid in the transcriptome of I. asprella root, including 72 u-nigenes for the upstream pathway and 26 unigenes for cyclization, oxidation and glycosylation in the downstream pathway. Phylogenetic analysis was carried out to further analyze the evolution relationship of some candidate uni-genes and their homologous genes. Two genes IaA S1 and IaA S2 were proved to be mixed amyrin synthases in yeast expression system. Moreover, IaA S1 was identified to one of the rare ASs with α-amyrin as the major product. It was concluded that a series of candidate genes, which might be involved in the biosynthetic pathway of triterpenoid saponins, were screened out from the transcriptome of I. asprella root. Further investigation of these candidate genes will provide insight into their actual functions in the triterpenoid saponins biosynthetic pathway in I. asprella.

ObjectiveTo investigate the expression change of cathepsin B (CathB) in the ventroposterior nucleus (VPN) of the ipsilateral thalamus after cortical infarction in rats.MethodsThe adult male Sprague-Dawley rats were randomly divided into either a sham operation group or a model group.The latter was further divided into postoperative 1-, 2-, 3-, 4-, and 8-week groups.A model of cerebral cortical infarction was induced by electrocoagulation the cortical branch of middle cerebral artery.Immunohistochemical staining and immunofluorescence were used to detect the protein expression and cellular localization of CathB in the VPN at each time point.ResultsThe expression level of VPN CathB in thalamus increased gradually after cerebral cortical infarction.It reached the peak at 4 weeks, and decreased at 8 weeks, however it was still higher than the control group (all P<0.05).The release of CathB from the lysosomes into the cytoplasm were found.In addition, the expression level of CathB in the activated astrocytes was significantly increased at 3 weeks after cerebral cortical infarction.ConclusionsDuring 1-8 week after cerebral cortex infarction, CathB in the VPN of the ipsilateral thalamus maintained higher expression level, suggesting that it might play a certain role in secondary degeneration in the thalamus after cerebral cortical infarction.

Aim To explore the effects of hydrocortisone on intracellular calcium in microglial cells.Methods The intracellular calcium was measured by instantaneous scanning with confocal laser microscope(CLM) in BV-2 cells, and fluo3-AM was used to dye the intracellular calcium.Results Both hydrocortisone and nicotine could obviously increase intracellular calcium in BV-2 cells(P<0.05).It was indicated by instantaneous scanning with CLM that hydrocortisone induced the rising of intracellular calcium immediately, and reached the peak about at the fifteenth second, and sustained for 10 seconds, then declined to baseline at 200th second.The effect of hydrocortisone on intracellular calcium exhibited a highly consistency with nicotine.Antagonist of glucocorticoid receptors RU486 could not abolish the rising of intracellular calcium induced by hydrocortisone(P>0.05);but the blocker of α7 nicotinic acetylcholine receptor(α7nAChR) methyllycaconitine could suppress the rising of intracellular calcium induced by hydrocortisone(P<0.05).Conclusion Hydrocortisone enhances intracellular calcium via α7nAChR in microglial cells, which not only demonstrates the non-genomic effect of glucocorticoid, but also suggests that glucocorticoid could serve as endogenous ligand of α7nAChR.