objective To determine the existence of virulence-associated invA gene in different genospeeies of Leptospira interrogans reference strains in China.and to understand the alterations of invA gene transcription and expression of L.interrogans strain Lai before or after infecting cells.Methods PCR was applied to detect the invA gene of four L.interrogans strains belonging to four different genospecies and L.biflexa strain Patoc Ⅰ.The entire invA genes from the L.interrogans strains were cloned and then sequenced.The prokaryotic expression system of invA gene of L.interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai was constructed.Using Ni-NTA affinity chromatography,the target recombinant protein rInvA was extracted and purified.Rabbits were immunized with rInvA to obtain antiserum and the titer of antiserum was determined by immunodiffusion test.A model of L interrogans strain Lai infecting human embryo kidney cell line HEK293 was established to detect the alterations of invA gene transcription and expression of the leptospiral strain before or after infecting the host cells by real-time fluorescent quantitative RTPCR and western blot assay.Results All the four tested L.interrogans strains had invA gene whereas L.biflexa strain Patoc Ⅰ not.The similarity of nucleotide and putative amino acid sequences of invA genes from the four L.interrogans strains belonging four different genospecies were 99.33%-100%and 98.66%-100%,respectively.The constructed prokaryotic expression system could efficiently express rInvA and the immunodiffusion titer of rabbit anti-rInvA serum was 1:16.After L.interrogans strain Lai infecting HEK293 cells for 30 min or above,the microbe could adhere the surface of the cells.On the 30 min after the infection,the mRNA level of invA gene of L.interrogans strain Lai was remarkably upregulated,and on the 45 min after infection the mRNA level presented a peak value and then graduated decreased.On the 45 min or 60 min after L.interrogans strain Lai infecting HEK293 cells,InvA protein could be detectable but before infection or after infection for over 90 min the InvA protein expression was negative.Conclusion The invA gene is a unique gene for pathogenic L.interrogans.The invA gene of L interrogans has characteristics of cell-touched expression and transient expression,which may have a close correlation with adhering and invading host cells.

Objective To explore the ultrasonographic appearance of dermatofibrosarcoma protuberans. Methods Clinical data, sonographic features and color Doppler flow imaging characteristics of 12 patients surgically and pathologically confirmed dermatofibrosarcoma protuberans were retrospectively analyzed.Results Single mass was detected in 11 patients (11/12, 91.67%) with the diameter ranged 0.9-8.0 cm. The tumors distributed in the trunk (8/12, 66.67%), proximal limbs (2/12, 16.67%) or face and neck (2/12, 16.67%). Most of the tumors (10/12, 83.33%) developed from skin and subcutaneous fat layer of the trunk and limbs and were hypoechoic and heterogeneous (11/12, 91.67%), some (8/12,66.67%) with distinct boundary and regular shape without liquification, calcification nor regional lymph node metastasis. Affluent color Doppler flow signals of tumor were found in 11 patients (91.67%).Conclusion Ultrasonographic characteristics of dermatofibrosarcoma protuberans include distinct boundary, regular shape, heterogeneous hypoechogenicity and rich color Doppler flow signals without regional lymph node metastasis.

Percutaneous coronary intervention (PCI) is becoming a common practice in the treatment of patients with coronary heart disease (CHD) of all age.Depression is considered to be a risk factor for the development of CHD and deteriorates the outcome after cardiac rehabilitation efforts.The aim of our study was to evaluate the presence of clinically relevant anxiety and depression in patients before and after PCI.Additionally we evaluated their relationship to age because of the increasing number of elderly patients undergoing PCI.Methods One hundred and twelve consecutive patients in three Sanatoria for Retired Cadres in Beijing who underwent PCI were asked to fill in the Hospital Anxiety and Depression Scale (HADS) to measure depression and anxiety scores two days before and ten days after PCI.Differences between these pre- and post-surgical scores were then calculated as means for changes,and the amount of elevated scores was appraised,in order to investigate the relationship between age and anxiety and depression,respectively,Spearman correlations between age and the difference scores were calculated.In addition,ANOVA procedures with the factor age group and McNemar tests were calculated.Results 25.8% of the patients were clinically depressed before and 17.5% after PCI;34.0% of the patients were clinically anxious before and 24.7% after PCI.This overall change is not significant.We found a significant negative correlation between age and the difference between the two time points for anxiety (Spearman rho = -.218,P = 0.03),but not for depression (Spearman rho = -.128,P = 0.21).ANOVA and McNemar tests revealed that anxiety scores and the number of patients high in anxiety declined statistically meaningful only in the youngest patient group.Such a relationship could not be found for depression.Conclusions Our data show a relationship between age and anxiety.Younger patients are more anxious before PCI than older ones and show a decline in symptoms while elderly patients show hardly any change.(J Geriatr Cardiol 2008;5:203-206)

Objective:To investigate the association patterns of chronic disease-relevant risk factors for the adults in Haidian District.Methods:Data for chronic disease-relevant risk factors for 3 219 adults in Haidian District in 2014 were collected and analyzed.SPSS 18.0 was used for statistical description and logistic regression.SPSS Modeler 14.1 was used to explore the association among the chronic disease-relevant risk factors.Results:Among men,5 patterns of chronic disease-relevant risk factors were identified,which suggested that heavy drinking,inadequate intake of fruit and vegetables,and physical inactivity were associated with smoking while inadequate intake of fruit and vegetables and smoking were associated with physical inactivity.Among women,one pattern of chronic disease-relevant risk factor was identified,which suggested that inadequate intake of fruit and vegetables was associated with physical inactivity.Conclusion:Chronic disease-relevant risk factors are intercorrelated among the adults in Haidian District.Information on patterns of chronic disease-relevant risk factors could assist interventions targeting multiple behaviors simultaneously.

Background Subretinal transplantation of retinal pigment epithelium (RPE) cells for the treatment of age-related macular degeneration (AMD) have accelerated the drive to develop xeno-free cultivation system that support the rapid differentiation of human embryonic stem cells (hESCs) into ES-RPE cells.Objective This study was to report a modified xeno-free culture system and method for accelerating derivation of hESCs to differentiate into RPE cells.Methods This study was approved by Ethic Committee of Zhongshan Ophthalmic Center.HESC H1 line was cloned and cuhured in Vitronectin XFTM-coated 6-well dish with xenogenetic-free medium.Cells were cultured in 50 ng/ml noggin,10 ng/ml DKK-1 and 10 ng/ml insulin like growth factor-1 (IGF-1) medium for 2 days,and then the concentration of noggin was decreased to 10 ng/ml and 5 ng/ml basic fibroblast growth factor (bFGF) and cultured for the following 2 days.Sequentially,noggin and bFGF were removed and cultured for 2 days.Finally,1 μmol/L CHIR99021 was added in medium for 6 days.Morphological changes in the progress of ESCs differentiation into RPE were observed by Living Cell Imaging System.The expression of Mitf and RPE65,RPE cellsspecific markers,in the cells were detected by immunofluorescence technique,and the relative expression levels of RPE cells-specific marker mRNA were assayed using real time fluorescent quantitation PCR.Results Polygonalshape monolayer cells which contained pigments were initially observed at day 14 after cultured with the cobblestonelike arrangement.Mitf and RPE65 were strongly expressed in the hES-derived RPE cells 35 days after induced,showing red fluorescence,and the cells presented hexagonal shape at cultured day 60 with numerous pigment granules in cytoplasm.Compared with before differentiation,the expression levels of Mitf mRNA in hES-RPE cells increased by (3.43±2.77) folds and (8.91 ± 2.83) folds,and the expression levels of RPE65 mRNA increased by (14.60 ± 3.94) folds and (87.16 ±9.32) folds at day 7 and day 14 after differentiation,respectively (all at P＜0.05).Conclusions A defined xeno-free culture system is successfully established by adding niacinamide,DKK-l,noggin,IGF-1 and CHIR99021 in xeno-free medium,and this system can accelerate the derivation and differentiation of hESCs into RPE-like cells.

Aim at the two problems in the field of traditional Chinese medicine (TCM) mechanism elucidation, one is the lack of detailed biological processes information, next is the low efficient in constructing network models, we constructed an auxiliary elucidation system for the TCM mechanism and realize the automatic establishment of biological network model. This study used the Entity Grammar Systems (EGS) as the theoretical framework, integrated the data of formulae, herbs, chemical components, targets of component, biological reactions, signaling pathways and disease related proteins, established the formal models, wrote the reasoning engine, constructed the auxiliary elucidation system for the TCM mechanism elucidation. The platform provides an automatic modeling method for biological network model of TCM mechanism. It would be benefit to perform the in-depth research on TCM theory of natures and combination and provides the scientific references for R&D of TCM.
Animals ; Automation ; instrumentation ; methods ; Databases, Factual ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Regulatory Networks ; Humans ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry

prognosis of patients with acute upper gastrointestinal bleeding (AUGIB). Methods A total of 130 patients with AUGIB in our hospital were enrolled in this study from August 2014 to August 2016 . Three kinds of scoring systems including Rockall, Blatchford and AIMS65 were used to evaluate the risk classification of the patients. Patients were followed up for 2 months. The incidence rates of re-bleeding and death within 2 months after admission were observed. Data of prognosis in patients with AUGIB were compared between the three scoring systems in the prognosis of patients with AUGIB, including the risk classification and the prediction accuracy of re-bleeding and death. Results There were no significant differences in the re-bleeding rate and fatality rate between the groups divided by Rockall and Blatchford scoring systems ( P>0.05). According to the grouping results of AIMS65 scoring system, the re-bleeding rate of low-risk group was lower than that in high risk group (1.45%vs. 13.11%, P=0.01), but there was no significant difference in fatality rate between the two groups (P>0.05). The values of area under the curve (AUC) of predicting re-bleeding rates by using Rockall, Blatchford and AIMS65 scoring systems were 0.6258, 0.6910, and 0.7241, and the values of AUC of predicting fatality rates were 0.7031, 0.7969, and 0.7031 by using receiver operating characteristic curve (ROC) analysis. There were no significant differences between them (P>0.05). The values of AUC of predicting re-bleeding rates by using the risk grading calculation by Rockall, Blatchford and AIMS65 scoring systems were 0.6189, 0.6139 and 0.7254 (P>0.05). But the values of AUC of predicting fatality rates were 0.6211, 0.6641 and 0.7695 (P<0.01). Conclusion The operation method of AIMS65 scoring system is simple and convenient, which is applicable to a wide range of patients with AUGIB. In the prediction of re-bleeding and mortality, AIMS65 scoring system has high accuracy and stability, which is worthy of promoting in clinical application.

Humans ; Infant ; Infant, Low Birth Weight ; Infant, Newborn ; Male ; Tyrosinemias ; diagnosis ; therapy

Objective To investigate the effect of suppressor of cytokine signaling 3 (SOCS3)on the proliferation of human mesangial cells stimulated by aggregated IgA1 (aIgA1) from patients with IgA nephropathy(IgAN),and explore its possible mechanism.Methods Serum monomeric IgA1 was isolated with jacalin affinity and Sephacryl S-200 HR chromatography from IgAN patients,and then heated to aggregated form (aIgA1).Human glomerular mesangial cells(HMC) were transfected with AdvSOCS3-IRES2-EGFP for 48 hours,and incubated with aIgA1 for 12-48 h.The cells were divided into blank control group,IgA1 group,IgA1 +Adv-EGFP group and IgA 1 +Adv-SOCS3-IRES2-EGFP group.The mesangial cell proliferation was observed through MTT,and the levels of SOCS3,TLR4,TGF-β1 protein and mRNA were detected through Western blotting and real-time PCR.Results HMC proliferation was promoted significantly after IgA1 stimulated at 24 h.Compared with control group,the protein and mRNA expression of SOCS3,TLR4,TGF-β1 were significantly increased in IgA1 group (P ＜ 0.05).Compared with IgA1 group and IgA1 +Adv-EGFP group,MTT absorbency was obviously reduced after incubation with aIgA1 for 24 h and 48 h in IgA+Adv-SOCS3-IRES2-EGFP group,and the protein and mRNA expression of TLR4 and TGF-β1 were significantly decreased in IgA1 +AdvSOCS3-EGFP group (P＜ 0.05).Conclusion Over-expression of SOCS3 may inhibit the proliferation of HMC stimulated by aIgA1,partly through down-regulating the expression of TLR4 and TGF-β1.

Objective To observe the effects of infrasound(IS)of different frequencies and intensities on the pathological morphology of the gastric mucosa(GM)in rats.Methods One hundred and forty male Sprague- Dawley rats were randomly and evenly divided into a control group(group A),an 8 Hz 90 dB group(group B),an 8 Hz 130 dB group(group C)and a 16 Hz 130 dB group(group D).IS with these frequencies and intensities was administered daily for 2 h to all groups except group A,which received sham infrasound.The other 70 rats were ran- domly and evenly divided into a second control group(group E)and an exposure group(group F),in which the rats were continuously stimulated by IS at 8 Hz and 130 dB for 2 h a day for 14 d.The pathological morphology of the GM in each group was observed at 1 d,7 d,14 d,21 d and 28 d after IS exposure.Results①Compared with group A,GM lesion scores were significantly increased in groups B,C and D at 1 d,7 d,14 d,21 d and 28 d(P＜0.01),but not in group B at 1 d.②Compared with group B,the GM lesion scores in group C were obviously in- creased at 1 d,7 d,14 d,21 d and 28 d,while scores were also obviously improved in group C in comparison with those in group D at 14 d,21 d and 28 d(P＜0.01).③The GM lesion scores in group F decreased gradually after IS,but were still higher than those in group E at 1 d,7 d,14 d,21 d and 28 d after IS.④The ultrastructures of the chondrosome and endocytoplasmic reticulum in GM cells were deformed after 8 Hz and 16 Hz IS.Conclusion 8 Hz 90 dB,8 Hz 130 dB and 16 Hz 130 dB IS can all result in GM damage in rats.The injury severity was closely related to the frequency,intensity and duration of the IS.Rats can adapt to IS after several exposures,and the damage tends to recover automatically.