To identify the cell wall-binding proteins of Streptococcus suis serotype 2(SS2), according to their structure feature, the substrates of sortase and I type secreted proteins with LysM domain, WxL domain, choline-binding domain,GW domain or S layer homologous domain in the recently published genome of SS2 strain 98HAH33 were firstly identified and then the putative functions were attributed to individual proteins by reference to the identification of conserved domains of InterPro and BlastP servers. Homologous proteins were identified by unfiltered BlastP homology searches (including conserved domain detection). Among the 23 putative proteins with a C-terminal LPXTG recognition signal for covalent attachment to peptidoglycan by sortase, 9 with I signal peptide were identified as sortase substrates. Among 9 substrates , YP_001201232, YP_001201531 and YP_001201656 had been experimentally verified to anchor to bacterial cell wall , and YP_001201232 known as the opacity factor of S. suis (OFS) was proved to be the virulence factors. According to function analysis, YP_001201484, YP_001201544, YP_001199825, YP_001197640, YP_001197840 and YP_001199755 appeared to be involved in SS2 pathogenesis. and YP_001200959, YP_001201233 and two proteins with LysM domain( YP_001199784 and YP_001201729) were the hypothetical proteins. These data suggest the majority of putative sortase substrates may implicate in the virulence of SS2 and could serve as a basis for targeted experimental studies into the function of these proteins.

Photosensitive skin diseases belong to physical skin diseases, which are caused by sunlight. Its clinical features are exposed parts of skin erythema, blisters or pleomorphic lesions, skin burning and itching consciously, and vary in different seasons. Traditional Chinese medicine believes that the pathogenesis of the disease lies in accountability phototoxic invasion, in vivo accumulation of heat, and natural tolerance to poor light. Traditional Chinese medicine therapy is often used in typing, syndrome differentiation, combination of traditional Chinese and Western medicine, Chinese medicine for external use and other means, and has achieved remarkable curative effects. However, there are still many problems about photosensitive skin diseases in the aspect of clinical and basic researches, and should be solved and improved.

Objective To develop cadres' stress scale for the Chinese people' s armed police forces.Methods Based on the stress theory and principles of the psychometrics,combined with characteristics of armed police forces. The cadres' stress scale was developed by ourselves. 802 cadres were evaluated as samples and statistic the data by item analysis ,factor analysis, reliability and valid analysis. Results The scale included four dimensions: task stress, economy stress, interpersonal stress and development stress. The internal consistency reliability was 0.893 ,the split-half reliability coefficient was 0.812. Retest reliability coefficient was 0.813. The criterion related validity to the stress scale and SCL-90 was good and the correlation coefficient with somatization, anxiety,depression, interpersonal sensitivity was 0. 376,0. 383,0. 396,0. 387 individually (P ＜ 0.05 ). Conclusion Cadres stress scale for cadres of Chinese people's armed police forces has good reliability and validity.

This study was aimed to compare the criteria on quality of questionnaire development and psychometric property evaluation. The references listed in the ISOQOL and consequent included criteria were searched. Two reviewers independently screened papers to include articles with inclusion criteria as follows: fully evaluating the questionnaire development and/or psychometric property; published or unpublished, but not book. Two reviewers independently extracted the information which contained criteria name, types of questionnaire assessed, criteria purpose, research methods, content, evaluation method and etc. Qualitative analysis was conducted to synthesize common characteristics of criteria, and to compare what each criterion emphasized on. The results showed that 9 included criteria sharing the same purpose which evaluated the questionnaire development process and/or psychometric properties of the health related quality of life questionnaire or patient reported outcome. Experts consensus based on previous studies were the criteria development methods. The standard content included conceptual framework, targeted population, content validity, internal consistency, test-retest reliability, structure validity and responsiveness with interpretability. Assessment scoring system used in the criteria was categorized as 2- or 4-point Likert scales. Or document of evaluation evidence was required to be provided. It was concluded that nine criteria can be used to develop new questionnaires, evaluate existed/new questionnaires, or choose the right questionnaires for clinic, based on the following elements such as their research purpose, trials types, questionnaire types or questionnaire administrations.

Aim To develop a sensitive,rapid and ac-curate LC-MS /MS method for the simultaneous deter-mination of cytochrome P450 probe substrates,inclu-ding caffeine and its metabolite paraxanthine for CYP1 A2,omeprazole and its metabolite 5-hydroxyome-prazole for CYP2C1 9,dextromethorphan and its metab-olite dextrorphan for CYP2D6,midazolam and its me-tabolite 1 ′-hydroxymidazolam for CYP3A4.Methods Probe drugs with the IS diazepam were extracted using ethyl acetate.Gradient elution was performed on an Agilent Eclipse Plus-C1 8 column (50 mm ×2.1 mm, 3.5 μm).The mobile phase consisted of 0.01 % for-mic acid(1 mmol·L -1 ammonium formate)and aceto-nitrile.The flow rate was 0.3 mL·min -1 ,and the in-jection volume was 1 0 μL.The analyte was detected u-sing electrospray ionization(ESI)in positive multiple reaction monitoring(MRM+)mode.The reaction se-lected ions were 1 95.0 /1 38.1 m /z for caffeine, 1 81 .1 /1 24.1 m /z for paraxanthine,346.1 /1 98.1 m /z for omeprazole,362.1 /21 4.1 m /z for 5-hydroxyome-prazole, 272.2 /1 47.1 m /z for dextromethorphan, 258.1 /1 57.1 m /z for dextrorphan,326.1 /291 .1 m /z for midazolam,342.1 /324.1 m /z for 1 ′-hydroxymid-azolam and 285.1 /1 54.0 m /z for diazepam as internal standard.Results The linear ranges of caffeine,pa-raxanthine,omeprazole,5-hydroxyomeprazole,dextro-methorphan, dextrophan, midazolam and 1 ′-hydroxymidazolam were 1 .95 ~2 000,0.98 ~250, 0.48 ~2 000,0.98 ~250,0.98 ~2 000,0.48 ~1 25,1 .95 ~2 000 and 1 .95 ~250 μg·L -1 respec-tively.The RSD of all probe drugs was less than 1 5%and matrix effects in plasma on the ionization of probe drugs were negligible.Conclusion This sensitive and rapid LC-MS /MS method is suitable for determination of the drug/metabolite concentrations in plasma,so as to study the metabolism of CYP1 A2, CYP2C1 9, CYP2D6 and CYP3A4 in depth.

To obtain and analyze the sequence of the nucleocapsid gene from bovine coronavirus, and to produce the fusion protein of the N gene in E.coli in order to use this recombinant protein for the study of bovine coronavirus. The N gene of BCV-DQ strain was amplified by RT-PCR, in which the primers were designed on the basis of N gene sequence of BCV-Mebus strain. The PCR products of 1 347 bp in length were cloned and sequenced, and then inserted into the prokaryotic vector pET30a. The recombinant plasmids were then transformed into Escherichia coli BL21 and identified by SDS-PAGE and Western blot assay. ELISA assay was optimized of N protein as the coating antigen to detect the viruses in the clinical samples. In comparison with 6 BCV strains in GenBank, the sequence identity was proved to be more than 98.3%. Result in SDS-PAGE showed that the fusion protein had a molecular weight of 60 ku, and could be specifically recognized by mouse serum against BCV. The indirect ELISA was used to test 256 serum samples collected from Heilongjiang province and 65.23% samples were positive. On testing field samples, an overall agreement of 95.31% was generated between the the neutralization test of viruses (VN) and indirect ELISA. It is apparent that the N gene was highly conservative and is expressed in E. coli in high level,also the prokaryotic expression products of this gene show a fine reactiongenicity in immune responses. It was also suggested that the N protein may be a useful antigen for sero-diagnosis and epidemiological investigation of BCV.

Objective To understand the risk factors of neurogenic pulmonary edema in the patients with severe hand foot and mouth disease(HFMD). Methods According to neurogenic pulmonary edema or not ,79 patients with severe HFMD were divided into two groups. The difference was analyzed on the clinical symptoms, signs,the outcomes of laboratory and electroencephalogram (EEG) examination between the two groups. Then the risk factors of neurogenic pulmonary edema was analyzed by logistic regression analysis.Results There were significant differences of the EV71 infection rate,high body temperature,myoclonia,limb weakness,the disability of eyeball regulation,tachycardia, hypertension or hypotension, the extension of capillary filling time, leucocytosist, creatine kinase isoenzyme, hyperglycaemia between two groups. However, there were no significant differences of fever, fever time, vomiting, somnolence, convulsion, limb tremor, c-reactive protein and EEG between two groups. Tachycardia, hypertension or hypotension, hyperglycaemia were significant risk factors for neurogenic pulmonary edema by logistic regression analysis. And hyperglycaemia was the most significant prognostic factor(odd ratio 27. 075, P = 0. 000 2). Conclusion Tachycardia, hypertension or hypotension,hyperglycaemia are the significant risk factors for neurogenic pulmonary edema. It is especially important for hyperglycaemia to predict neurogenic pulmonary edema.

Objective: To investigate the properties of radiation-induced changes of cell cycle and apoptosis in breast cancer cell lines MDA-MB-231 and MCF-7, and to further explore its relationship with radiosensitivity. Methods: The two cell lines MCF-7 and MDA-MB-231 were irradiated with 6 MV X-rays. The cell survival curves were fitted by clonogenic formation assay, then according to the radiosensitivity parameters average lethal dose (D₀), quasi-threshold dose (D_q) and survival fraction after 2 Gy irradiation (SF₂), the radiosensitivity of the two cell lines was compared. The apoptosis rate and cell cycle distribution of the two cell lines were detected by Hoechst 33342 and PI staining. Results: The survival curve of MDA-MB-231 cell line shifted to the right compared with that of MCF-7 cell line. The values of D₀, D_q and SF₂ of MDA-MB-231 cell line were higher than those of MCF-7 cell line (1.603±0.023 vs. 1.233±0.027, 1.76±0.04 vs. 1.03±0.10, 0.639 3±0.008 2 vs. 0.398 1±0.018 5, t values were -17.981, -11.745, -20.596, P values were 0.000, 0.003, 0.000). The apoptosis rate of MCF-7 cell line was significantly higher than that of MDA-MB-231 cell line at the doses of 2, 4, 8 Gy irradiated 24 h and at the 12, 48 h after 6 Gy X-irradiation (t values were 4.441, 7.299, 10.499, 6.375, 7.743, P values were 0.011, 0.002, 0.000, 0.003, 0.001). Compared with the non-irradiated group, G₂ phase arrest appeared in both cell lines after irradiation. The percentage of the G₂/M phase in MDA-MB-231 cell line increased as the time or dosage accumulated. Furthmore, the percentage didn't go down even after 48 hours later. However, the blockage began to gradually release in MCF-7 cell line at the dose of 8 Gy irradiated 24 h and the 48 h after 6 Gy X-irradiation. Followed with that, it turned out the percentage of the G₀/G₁ phase increased [(65.80±0.56)%, (62.53±0.67)%]. Conclusions 6 MV X-irradiation with the doses of 2-8 Gy can induce the cell cycle arrest at G₁ and G₂ phase in MCF-7 cell line, G₂ phase in MDA-MB-231 cell line. Thus more apoptosis appears in MCF-7 cell line, which may cause the difference in radiosensitivity between the two cell lines.

AIM: The aim of this work is to investigate the protective effects of Ginsenoside Rb 1(Rb 1) on apoptosis induced by hypoxia /hypoglycemia and reoxygenation in cultured rat hippocampal neurons. METHODS: Apoptosis were measured by flow cytometry; Morphological changes and neuronal necrosis were examined under microscope; The leakage of lactic dehydrogenase(LDH) and the product of nitric oxide(NO) were measured. RESULTS: hypoxia /hypoglycemia cultures for 5 h and reoxygenation induced neuronal apoptosis and necrosis,and significantly increased the leakage of LDH and the product of NO. The effects were enhanced with the extending time of reoxygenation. Rb 1 could significantly decrease the percentage of neuronal apoptosis and necrosis, and reduce the leakage of LDH and the product of NO. CONCLUSION: Rb 1 had an effect of anti-neuronal apoptosis. This effect might be related to the inhibition of the activity of NO synthase and NO production.

BACKGROUND:Shengjiyuhong ointment has been reported to inhibit hypertrophic scarring. OBJECTIVE:To verify the effects of Shengjiyuhong ointment on hypertrophic scarring of in a rabbit ear model. METHODS:Each ear of thirty-six Japanese rabbits was used to make four 1-cm-diameter circular ful-thickness skin wounds with the entire perichondrium removed. Final y, 288 wounds were made and randomly divided into 6 groups:model, negative control (no drugs were administered), low-, moderate-, high-crude herbal dose drugs (Shengjiyuhong ointment was administered topical y at concentrations of 8.39%, 25.18%, and 75.54%), and positive control (recombinant bovine basic fibroblast growth factor was administered topical y). Shengjiyuhong ointment was administered twice daily til wound healing. The wounds were evaluated by the Vancouver scar scale (VSS). Scar elevation index (SEI) of scar specimens was calculated under a microscope at 40× magnification. mRNA expression levels of type I and III col agen, connective tissue growth factor, fibronectin, andα-smooth muscle actin (α-SMA) were determined by fluorescent quantitative PCR. Protein expression levels of type I and III col agen andα-SMA were detected by western blot assay.α-SMA immunoreactivity was determined by immunofluorescent staining. RESULTS AND CONCLUSION:VSS scores and SEI were significantly increased in each group at 30 days (P<0.05). VSS scores and SEI were significantly decreased in the moderate-and high-crude herbal dose drug groups and positive control groups compared with the model, negative control, and low-crude herbal dose drug groups (P<0.05 or P<0.01). mRNA expression levels of type I and III col agen, connective tissue growth factor and fibronectin, and protein expression levels of type I and III col agen andα-SMA were significantly inhibited after moderate-crude herbal dose Shengjiyuhong ointment and positive drug treatment (P<0.01). These findings suggest that Shengjiyuhong ointment can reduce hypertrophic scars by inhibiting fibroblast proliferation and col agen deposition.