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.

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.

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

Objective To investigate the expression of Nogo-66 receptor (NgR) in primary cultured rat astrocytes.Methods RT-PCR and Western blot were applied to verify mRNA and protein expression of NgR in primary cultured and purified astrocytes. Indirect immunofluorensence and confocal microscopic technique were used to study the distribution of NgR. Results Specific NgR cDNA product could be amplified from the total RNA of primary cultured astrocytes by RTPCR; Western blot of the extracts of astrocytes demonstrated a specific NgR band at about 64 kD. Indirect immunofluorescence and confocal scanning further revealed the intracellular localization of NgR protein in astrocytes. Simultaneously,the NgR protein was detected in C6 rat glioma cells by western blot and immunofluoresence staining. Conclusion NgR is expressed in primary cultured astrocytes, which offers strong and direct support for the expression of NgR in astrocytes in vivo.

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.

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 investigate the expression of CXCR4 in cultured human dental pulp cells (HDPC) in vitro and the corresponding ligand SDF-1α level of HDPC supernatants stimulated by lipopolysaccharide(LPS)and tumor necrosis factor-α(TNF-α),and to explore the role of SDF-1αon the proliferation and the migration of HDPC.Methods The expression of CXCR4 in HDPC was detected by immunocytochemistry technique and indirect immunofluorescence technique.The culture supernatants of HDPC were collected after HDPC had been simulated by LPS and TNF-α of difierent concentrations for 48h and then the SDF-1α level was assayed by quantitative sandwich ELISA.Meanwhile,the effects of recombinant human SDF-1α(rhSDF-1α)on the proliferation and the migration of HDPC at different concentrations were observed by MTT and Boyden Chamber Assay.Results CXCR4 was expressed in cytomembrane of HDPC and SDF-1α was secreted into their normal cell supematants with a concentration of(4513.55±962.92)ng/L. The secretion of SDF-1α was both significantly decreased by stimulation with LPS and TNF-α(P＜0.05).In addition,rhSDF-1α stimulated the HDPC proliferation at the concentrations of 50,100,200μg/L(P＜0.01)and increased the chemotactic migration of HDPC significantly after 9h's incubation with the concentrations of 50,100μg/L(P＜0.05).Conclusions SDF-1α accelerated the proliferation and the migration of HDPC which expressed CXCR4.SDF-1-CXCR4 axis may play a role in repair of pulp injury.

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.