Objective:To investigate whether His-tag to change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.Methods:The G1 and F/M2 gene fragments were amplified by PCR method and then ligated into the expressing vector pET-His or pET-DsbA-His.Each recombinant plasmid was transferred into E.coli BL21(DE3) and the expression was induced by IPTG.The expressed His-G1F/M2 or DsbA-His-G1F/M2 was purified by affinity chromatography.The latter was digested with thrombase and G1F/M2 was purified by affinity chromatography.His-G1F/M2 or G1F/M2 was used to immunize BALB/c mice.Anti-RSV antibody was measured by ELISA and RSV-specific CTL responses by MTT.Results:No significant difference was observed between the level of anti-RSV antibody or RSV-specific CTL response induced by G1F/M2 and that by His-G1F/M2.Conclusion:His-tag does not change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.

In the basis of a large amount of literatures, this article sumed up the characteristics and application of eggshell membrane.

Four salts of ticagrelor, ticagrelor-3,5-dinitrobenzoic acid, ticagrelor-pyrazinamide, ticagrelor-D-proline and ticagrelor-L-proline were prepared by solvent suspension and liquid-assisted grinding to improve the solubility of ticagrelor. The compounds were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, elemental analysis, and the intermolecular salt-bonding forces were analyzed. The equilibrium solubility of salts and pure drug in hydrochloride buffer pH 1.2 and phosphate buffer pH 6.8 were measured by high-performance liquid chromatography. Ticagrelor was salted with 3,5-dinitrobenzoic acid, pyrazinamide, D-proline, L-proline all in a stoichiometric ratio of 1∶1; with the exception of ticagrelor-D-proline, the solubility of the other three salts provided significantly improved solubility in hydrochloride buffer pH 1.2, and the equilibrium solubility of ticagrelor-3,5-dinitrobenzoic acid was increased by approximately 1.7 folds as compared to pure drug. Salt-forming technology is convenient and can improve the solubility of ticagrelor.

The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer, the physical status of HPV-16 in patients with cervical lesions, and the role of HPV-16 integration in cervical carcinogenesis were investigated. HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women. Multiple PCR was done to detect physical status of HPV-16 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection, including 112 specimens with cervical cancer, 151 specimens with CIN I, 246 specimens with CIN and 120 specimens with CINIII. The results showed that there were 112 cervical cancer samples (56.57% of total cervical cancer patients) with HPV-16 infection. The frequency of HPV-16 pure integration was 65.18% (73/112), 56.57% (47/120), 23.58% (58/246) and 7.95% (12/151) in cervical cancer, CINIII, CINII and CINI patients respectively. In situ hybridization was performed on some paraffin-embedded sections of CINII, CINIII and cervical cancer to verify the physical status of HPV-16 infection. Significant difference was observed between cervical cancer and CIN I, CINII, CINIII in the frequency of HPV-16 integration (P<0.01). It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer. In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration. The application of testing HPV genotyping and physical status based on detection of HC-II HPV DNA would be in favor of predicting the prognosis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.

Piroxicam has polymorphism. Different crystalline forms can exhibit different physicochemical properties and biological activities. Analysis of the intermolecular interactions is essential to reveal the formation mechanism and differences of polymorphs. In this paper, Hirshfeld surface analysis and semi-empirical methods were used to calculate and analyze the intermolecular interactions in seven polymorphic forms of piroxicam. The results show that the Hirshfeld surface analysis method can clearly and intuitively reveal the intermolecular interactions, among which H…H, O…H/H…O and N…H/H…N interactions account for 95% of the total energy. There are differences in the proportion and distribution of the forces of different crystal forms. The energy calculation shows that the lattice energy of the hydrate is significantly lower than that of the anhydrous forms, and in the specific energy distribution, the contribution of the dispersion force is the most prominent. Further interaction energy analysis was found that within the distance of 3.8 Å from the center of the piroxicam molecule, different crystalline forms of piroxicam molecule have different interaction energies with surrounding molecules.

Objective To observe the effects of mycophenolate mofetil (MMF) on the differentiation and proliferation of Helper T cells 17 (TH 17),so as to reveal its role and the possible mechanism in inducing immunological suppression.Methods Sixteen Balb/c mice of SPF level aged 8 weeks were randomly divided into two groups:MMF group and control group,with 8 mice in each group.In MMF group,the mice received intragastric administration of MMF (40 mg·kg-1· day-1 ),and those in control group received intragastric administration of identical volumetric saline every day.After three weeks,peripheral blood was collected and spleen cells were prepared.Flow cytometry was used to determine the proportions of CD4+ TH 17 and CD4+ CD25+ Tregs,then the ratio of TH 17/Tregs was calculated,and the concentrations of interleukin-1 7 (IL-1 7) and interleukin-23 (IL-23) in serum were measured by ELISA.Results The proportion of CD4+ TH 17 in the peripheral blood and spleen was (1.95 ± 0.08) and (2.42 ± 0.06) in MMF group,and (3.19 ± 0.07)％ and (4.21 ± 0.25)％ in control group,respectively.There were significant differences between the two groups (P ＜0.05).Meanwhile,the ratio of TH 17/Tregs in MMF group,both in the peripheral blood and spleen,was significantly decreased as compared with the control group (P＜0.05).The concentration of IL-17 in MMF group was lower,but that of IL-23 in MMF group was higher than in the control group (P＜0.05).Conclusion MMF could obviously suppress the differentiation and proliferation of CD4+ TH 17 in vivo,reduce the ratio of TH17/Tregs and the IL-17 secretion,thus facilitate the induction of immune tolerance.

Nasal polyp (NP) is a common chronic inflammatory disease of the nasal cavity and sinuses.Although some authors have suggested that NP is related to inflammatory factors such as interleukin (IL)-1β,IL-5,IL-8,granulocyte-macrophage colony-stimulating factor (GM-CSF),tumor necrosis factor (TNF)-α,and IL-17,the mechanisms underlying the pathogenesis and progression of NP remain obscure.This study investigated the expression and distribution of IL-17 and syndecan-1 in NP,and explored the roles of these two molecules in the pathogenesis of eosinophilic chronic rhinosinusitis with nasal polyps (Eos CRSwNP) and non-Eos CRSwNP.Real-time PCR and immunohistochemistry were used to detect the expression of IL-17 and syndecan-1 in samples [NP,unciform process (UP) from patients with CRS,and middle turbinate (MT) from healthy controls undergoing pituitary tumor surgery].The results showed that the expression levels of IL-17 and syndecan-1 were upregulated in both NP and UP tissues,but both factors were higher in NP tissues than in UP tissues.There was no significant difference in IL-17 levels between the Eos CRSwNP and non-Eos CRSwNP samples,and syndecan-1 levels were increased in the non-Eos CRSwNP tissues as compared with those in Eos CRSwNP tissues.In all of the groups,there was a close correlation between the expression of IL-17 and syndecan-1 in nasal mucosa epithelial cells,glandular epithelial cells,and inflammatory cells,suggesting that IL-17 and syndecan-1 may play a role,and interact with each other,in the pathogenesis ofnon-Eos CRSwNP.

Objective To investigate the attitudes of urban and rural community members toward total banning on smoking in public places and to explore the factors associated with these attitudes, in three counties/cities in China. Methods A cross-sectional study was conducted in three counties/cities in 2004, including Xin' an county of Henan province, Anyi county of Jiangxi province, and Mianzhu city of Sichuan province. A total of 5642 residents at age of 18-69 years old were interviewed face-to-face with a uniform questionnaire by locally-trained interviewers, through a random three-stage stratified sampling in each county. Factors were assumed to be associated with attitudes, using chi-square test in univariate analysis and non-conditional logistic regression model in multivariate analysis. Results The prevalence of current smoking among respondents was 44.3%. 80.7% (1379/1709) of the current smokers reported smoking often or sometimes in public places. Only 9.6% (479/4983) of the respondents reported that their indoor workplaces had totally banned on smoking. 43.5% of the respondents supported a total smoking ban strategy in pubic venues. The results of multivariate logistic regression model showed that eight factors were significantly associated with support for the total smoking ban in public places included region, residency, age, gender, education, smoking status, awareness about passive smoking hazards, and hearing of any message on tobacco control through media differences of rates regarding the factors as: residents in urban to rural areas (OR=1.29), elderly to youngsters (30-49 vs. 18-29, OR=1.46; 50-69 vs. 18-29, OR=1.71), female to male(OR=1.27), high-educated to less-educated ones, quitters to current smokers (OR=1.90), nonsmokers to current smokers (OR=2.01). Those who know messages on health hazards of passive smoking (OR=2.26), or heard of message on tobacco control through media (OR= 1.43). Conclusion Results from our study revealed that a thorough smoke-flee policy in public places should be developed and implemented in these three counties/cities.

OBJECTIVE: To establish 2-dimensional electrophoresis (2-DE) maps of Helicobac-ter pylori in human gastritis, and gastric cancer, to identify the differentially expressed proteins,and to discuss the role of bacterial factor in pathogenesis. METHODS: The total proteins of Helicobacter pylori in human gastritis and gastric cancer were separated by immobilized pH gradient-based 2-DE. The differentially expressed proteins were screened by PDQuest analysis software and identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry, and searched on database. RESULTS: A well-resolved and reproducible 2-DE pattern of Helicobacter pylori was obtained from patients with human gastritis and gastric cancer. Fourteen differentially expressed proteins were identified, including proteins related to anti-oxidation,molecular chape-rones and detoxification, enzymes related to metabolism,proteins related to cytoarchitecture,and proteins related to signal conduction. CONCLUSION A well-resolved and reproducible 2-DE pattern of Helicobacter pylori in human gastritis and gastric cancer is established and differentially expressed proteins from these 2 diseases are identified. The differentiation of protein expression may play an important role in the pathogenesis of gastric cancer.
Bacterial Proteins ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Female ; Gastritis ; microbiology ; Helicobacter Infections ; microbiology ; Helicobacter pylori ; Humans ; Male ; Proteome ; analysis ; Proteomics ; methods ; Stomach Neoplasms ; microbiology

Objective:To observe the application effect of formative evaluation of embedded standard patient (SP) on internal medicine practice teaching.Methods:In this study, 110 students from our school in Batch 2014 who were interned in the internal medicine of the same affiliated hospital were selected as the research objects, and they were randomized into the experimental group and the control group, with 55 students in each group. The control group was trained with traditional clinical practice teaching methods. The experimental group used SP combined mini-clinical evaluation exercise (Mini-CEX) to participate in the evaluation during the clinical practice teaching of internal medicine. After the internal medical practice, the two groups of students were evaluated by Mini-CEX, and the teaching effect of both groups was evaluated through questionnaires.Results:There was no significant difference in the degree of difficulty in the examinations ( P>0.05), but the test scores of the experimental group were significantly higher than those of the control group. The results of Mini-CEX used in inquiry skills, physical examination, humanistic care, and organizational effectiveness were significantly higher in the experimental group than in the control group. The experimental group was highly satisfied with the formative evaluation of SP participation in improving the effect of internal medicine practice, with statistical significance ( P<0.05). Conclusion:The formative evaluation with SP participation in this study can improve the clinical practice teaching effect of medical students to a certain extent.