OBJECTIVE: To establish spectrophotometry and HPLC methods for the determination of ferrous succinate tablet and to compare the results of above two methods with that of cerium sulphate method.METHODS: Ferrous reacted with 1,10-phenanthroline to form colored complex and spectrophotometry was applied to measure its absorbance.The detection wavelength was set at 510 nm.HPLC method was performed on C18 column with mobile phase consisted of 0.05 mol?L-1 KH2PO4 (pH=2.5) at detection wavelength of 215 nm.The method was quantified by an external standard mode.RESULTS: The linear range of ferrous in spectrophotometry was 0.042 44～2.546 4 mg?L-1 (r=0.999 8) with an average recovery of 99.5% (RSD

ObjectiveTo investigate the adjuvant effect of heatshock protein 110(HSP110) on the immune responses induced by an altered peptide ligand of human papilloma virus type 16 E711-20 peptide (HPV16E711-20).Methods The complex of HSP110 and an altered peptide ligand of HPV16E711-20 was constructed in vitro.Fifteen 6-week-old C57BL/6 female mice were randonly and equally divided into 3 groups,including complex group,ligand group,and phosphate buffered solution (PBS) group,to receive intraperitoneal immunization with the complex (100 μg),peptide (10 μg),and PBS (100 μl) respectively.Immunization was carried out with an interval of 2 weeks for 2 times.Two weeks after the last immunization,the mice were sacrificed followed by the isolation of splenocytes.MTT assay was performed to evaluate the proliferation activity of splenocytes,intracellular staining for interferon(INF)-γ to detect cytotoxic T lymphocytes (CTLs),standard chromium-51 (51Cr) release assay to estimate the lethal effect of specific CTLs on target cells.Statistical analysis was performed by using t test with SPSS 10.0 software.Differences were considered as statistically significant when the P value was less than 0.05.ResultsA significant increase was observed in the proliferation index ( 1.87 ± 0.122 vs.0.32 ± 0.071,t =4.01,P ＜ 0.01 ) of,and percentage of CD8+IFN-γ+ T lymphocytes(3.9％ vs.0.4％,t =3.88,P ＜ 0.01 ) among splenocytes from the complex group compared with the ligand group.At the effector-to-target ratio of 100 ∶ 1,50 ∶ 1,25∶ 1 and 12.5 ∶ 1,the death rate of target cells was 54.7％,72.2％,61.5％ and 39.8％ respectively after incubation with CTLs from the compleximmunized mice,higher than that from the ligand-immunized mice (35.2％,49.3％,28.1％,17.4％,respectively).ConclusionHSP110 could enhance the immunological effect of the altered peptide ligand of HPV16E711-20,and can serve as an immunological adjuvant.

Objective To analyze the difference of salivary microbiome between the patients with functional dyspepsia and healthy controls.Methods Saliva samples were collected from the patients with functional dyspepsia and healthy control.Genomic DNA of the samples was extracted,and the high-throughput sequencing technology was used to conduct DNA sequence of 16S rRNA-V4 region.Subsequently,all the data were performed by the bioinformatic analysis.Results The salivary microbiome in the functional dyspepsia group was dominated by Proteobacteria,while Bacteroidetes was the top microbiota in the heathy control group.In the functional dyspepsia group,the Chao1 index,ACE index,Shannon index and Simpson index were 1 295,1 351,4.93 and 0.90 respectively.In the healthy control group,the above indexes were 1 001,1 351,5.28 and 0.92 respectively.The PCoA ba sically separated the microbiome composition of the two groups.Sixteen kinds of microbiota were significantly different between two groups using linear discriminant analysis effect size tool,including Bacteroidetes,Prevotella,Prevotellaceae,Neisseria,Betaproteobacteria and Proteobacteria,etc.Conclusion Saliva in the patients with functional dyspesia has characteristic microbiome composition,which is dominated by Proteobacteria,but the richness and diversity between the two groups have no difference.Neisseria is one of the significantly different bacteria between the two groups.

ObjectiveTo assess the relationship between the estimated glomerular filtration rate (eGFR) and the severity of coronary artery stenosis in elder people. MethodsSix hundred and fifty-seven patients who underwent selective coronary artery angiography were divided into 3 groups according to age: group Ⅰ (202 patients aged <60 years), group Ⅱ (181 patients aged from 60 to 69 years) and group Ⅲ (274 patients aged ≥70 years). According to the simplified MDRD equation,impaired renal function and severe renal dysfunction, respectively. Clinical features and the severity of coronary artery stenosis were compared among the 3 groups. ResultsThe level of average eGFR tended to he decreased with the narrowed coronary arteries. The difference of average eGFR between patients with muhivessel versus two-vessel coronary artery disease was of statistical significance in Group Ⅲ(P = 0. 035). After controlling for the effects of smoking, hypertension, dyslipidemia,aging, and diabetes, the Gensini score which reflected the severity of coronary artery stenosis was negatively correlated with the level of eGFR (r =-0. 110, P = 0. 005). ConclusionsIt suggests that the eGFR decrease is associated with the severity of coronary artery stenosis, and there is a linear correlation between eGFR and Gensini score in elder people, eGFR can be used to evaluate the severity of coronary artery disease and prognosis.

Objective To assess the characteristics and satisfaction of outpatients in walk-in clinic of a grade A tertiary hospital.Methods A total of 1 100 outpatients visiting walk-in clinic of the Second Hospital of Hebei Medical University during March 2016 and June 2016 were randomly selected for the questionnaire survey.The self-designed questionnaire included the general information of patients,the satisfaction degree to the walk-in clinic,to special clinics of the hospital or to community clinics and related reasons,and suggestions for further improvement.Results Total 947 valid questionnaires were received with a retrieval rate of 86.09％.Most of participants had one chronic disease (594,62.72％).The satisfaction degree of walk-in clinic (1.78) was higher than that of hospital special clinics (1.93) (P =0.010),in turn the latter(2.33)was higher than that of conmmnity clinics (P ＜ 0.001).The reasons for patient to choose the walk-in clinic were wide range of medications (389,41.08％),reliable quality (285,27.98％),short waiting time (249,26.29％) and so on.Suggestions for improvement of the walk-in clinic were further reducing the waiting time (428,45.20％),need specialists for chronic disease management (210,22.18％),raising reimbursement rate (172,18.16％) and so on.The reasons for patients to choose special clinic were need readjusting prescription (587,61.99％),special disease or drug (427,45.09％).The reasons of dissatisfaction with special clinic were long waiting time (476,50.26％),frequent transfer to multiple departments (221,23.34％),high registration fee (221,23.34％).The reasons for patients to choose community clinic were short distance (350,36.96％),better environment (350,36.96％),short waiting time (184,19.43％).The reason of dissatisfaction with community clinic were low qualification of medical staff (405,42.77％),insufficient medications (392,42.39％);distrusting the quality of drugs (263,27.77％).Suggestions for improvement of the community clinics were upgrading the qualification of medical staff (317,33.47％),enlarging drug variety (298,31.47％),improving medical environment (94,9.93％) and so on.Conclusion Most outpatients visiting walk-in clinic of grade A tertiary hospital are highly satisfied the clinic.To meet the patients' needs the primary medical unit should further improve their overall service.

Exocyst complex is known to function in the exocytosis network, however, the molecular mechanism is unclear yet. Using UV/trimethylpsoralen mutagenesis, the sec-10 (one component of the exocyst complex) knockout mutant of C. elegans was obtained for the first time. The drug sensitive assays revealed clearly that the sec-10 gene affected the neural signal transmission, however, the electrophysiological assay showed the function of the ionotropic receptors in the neuromuscular junctions (NMJs) were unaltered compared with the wild type (WT). Thus it was assumed that the sec-10 gene might not influence the known ionotropic receptors in the NMJs, but some other pathways instead.

Mimecan belongs to a family of leucine-rich proteoglycans that are secreted into the extracellular matrix. In order to investigate the function of mimecan, the coding region of mimecan was amplifed from a human pituitary cDNA by PCR and the recombinant prokaryotic expression vector pGEX-M was constructed. The vactor was transformed into E.coli BL21(DE3)and the GST-M fusion protein of 38 Kd was ecpressed in the bacteria under induction of IPTG. After purification, the fusion protein was infucted into New Zealand rabbits to prepare polyclonal antibody. The antibody was tested by Western blotting for their specificity and sensitivity. Using the antibody it was found the mimecan was expressed highly in certain types of human pituitary tumor tissues. These results make it possible for studying the biological function of mimecan.

Objective To observe expression of resistin gene in adipose tissue of SD rats fed with high-fat diet and its correlation to insulin resistance, diabetes mellitus and leptin. Methods Using RT-PCR to mRNA from adipose tissue, the products were obtained, purified, constructed and squenced. Once its squence was verified, semi-quantitative RT-PCR and Northern Blot were employed to study its expressions in adipose tissues of rats with different diets. Oral glucose tolerance test and insulin release test were performed in 2 groups of rats. Results (1) The resistin gene was successfully constructed and sequenced, which was in accordance with the data of genbank. (2) Compared with the control, the expression of resistin in adipose tissue of high-fat diet rats was significantly enhanced, and blood glucose, serum insulin and leptin levels were also elevated in high-fat diet rats (P

Objective To observe the effect of berberine on glucose transport in 3T3-L1 adipocytes and to investigate its mechanism. Methods The glucose consumption of the cells was determined by the glucose oxidase method. The glucose transportation rate of the cells was assayed by the uptake of 2-deoxy-〔 3H〕-D-glucose. Protein kinase B (Akt) activity was detected by immunoprecipitation and Western blot. The gene expression of c-Cbl-associated protein (CAP) was detected by Northern blot. Results 0.1～200 ?mol/L berberine significantly increased glucose consumption in 3T3-L1 adipocytes with a dose-dependent effect, which was independent of insulin. The glucose transportation was significantly increased in adipocytes incubated with 0.1～10 ?mol/L berberine; the action began at 2 h and reached a peak value at 12 h. The results of immunoprecipitation and Western blot showed that berberine did not enhance Akt activity. The result of Northern blot indicated that berberine significantly decreased CAP mRNA expression. Conclusion Adipocytes are the important target cells of berberine. Berberine significantly increases glucose transportation and consumption in adipocytes, the action appeares to be independent of insulin signal pathway.

Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.