Objective To evaluate the prospects of recombinant 38000 protein of Mycobacterium tuberculosis in tuberculosis epidemic investigation and subunit vaccine preparation.Methods Physicochemical characteristics of recombinant 38000 protein was detected by P I, peptide-mapping analysis and circular dichroism,guinea pig skin test,MTT stain,and peripheral blood macrophage phagocytosis were used to investigate the roles of recombinant 38000 protein in the cell immunity.Results Recombinant 38000 protein was acidic protein,its P I, was 4 67.The number of alkaline amino acid correspond with theoretic number;The secondary structure of recombinant 38000 protein was composed of ?-helix(32 6%),?-turn(31 6%) and random coil(35 8%) Recombinant 38000 protein could induce DTH in guinea pig sensitized by Mycobacterium tuberculosis Recombinant 38000 protein enhanced phagocytosis of macrophage in mice . PBMC from 30 8% healthy donors and 25% tuberculosis patients were stimulated by the recombinant 38000 protein.Conclusion Recombinant 38000 protein may be used as diagnostic reagent and as an candidate in development of subunit vaccine.

Objective To compare expression of extracellular proteins of virulent H37Rv and attenuated H37Ra in order to search differential proteins,to provide a train of thought for studing M.TB toxicity further.Methods Extracellular proteins were extracted from H37Rv and H37Ra which were inoculated and cultured on Suton's medium for three weeks.The first dimensional electrophoresis was performed on immobilized pH gradient rod gels(pH 3～10).Then the proteins in the rod gels were separated using SDS-PAGE gels.The silver-stained gels were dried and scanned with image scanner.The 2D image analysis was performed with image Master 2D Elite 3 10.Results The most protein spots deriving from extracellular proteins of H37Rv and H37Ra strains were in acidic range.In the basic range(pI more than 9 0),the number of protein spots belong to extracellular proteins of H37Rv and H37Ra was few.Three protein spots belong to low molecular range in H37Rv strain.However,absent in H37Ra strain.Conclusions Two-dimensional gel electrophoresis is useful to separate protein in Mycobacterium tuberculosis.

Objective To understand the rpoB gene mutation in M.tuberculosis isolates,and to evaluate their clinical value.Method 335 clinical isolates of mycobacterium tuberculosis(109 isolates drug susceptible,246 isolates rifampin-resistance or multidrug resistance including rifampin) were detected using polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP).Results SSCP pattern of reference mycobacterium tuberculosis H37Rv as control,no mutation was found in rifampin-suscepitible 109 strains.SSCP patterns of 225/246 rifampin resistant clinical isolates were different from the normal control.The sensitivity was 91 5%.31 resistant isolates,included 25 abnormal isolates and 6 normal isolates of SCCP were identified.Sequencing showed 29 isolates had rpoB gene mutations and 3 isolates were not found rpoB gene mutations.The 3 most frequent rpoB gene mutation situs were Leu-531(19 isolates.TCGTTG) and His-526(7 isolates,CACTAC) and Asp-516(3 isolates,GACGTC).Conclusions The results confirm that rpoB gene mutation is the most important mechanism in rifampin resistant tuberculosis mutation situs are 531 tryptophan and 526 histidine;respectively.It is feasible that using PCR-SSCP to detect drug resistance in mycobacterium tuberculosis.

Under the new situation of rapidly expanded medical education scale and its great development,combined with the characteristics and position of higher medical education, and keeping the quality of teaching in this school lifeline,we explore the specific measures and methods to improve the quality of medical education, from the quality monitoring system of classroom teaching,clinical teaching and teaching management.

Objective:To research whether ectopic over-expression of Pim-2 could cause chang-liver cell (LO2) malignant transformation,to explore the relationship between Pim-2 protein and hepatocellular carcinoma.Methods:Three groups of cells were arranged including human chang liver cell line LO2 (group C),LO2 cells transfected with empty-vector (group B) and LO2 cells transfected with Pim-2 gene (group A).Pim-2 expression levels were detected.The morphology,proliferation level,apoptosis rate and migration ability of the cells were detected respectively.The cells were subcutaneously inoculated into athymic mice and the microstructures of the neoplasm were observed by optical and electron microscopy.Results:Compared with group B,Pim-2 expression levels were significantly higher in group A (P＜0.05),and their morphology had obvious malignant changes.They also showed a significantly increased proliferation rate (P＜0.05) and migration capacity (P＜0.05),as well as a significantly decreased apoptosis rate (P＜0.05).Only the athymic mice inoculated with group A could generate neoplasm,and the morphology of the neoplasm coincided with that of the hepatoma.Conclusion:Both the morphological and biological changes of LO2/Pim-2 cells indicate the trend of malignant transformation,which could generate hepatoma in athymic mice.Pim-2 could induce malignant transformation of human liver.

OBJECTIVE:To establish a method for the content determination of paeoniflorin in Bazhen pill(concentrated pill). METHODS:HPLC was performed on the column of wondasil C18 with mobile phase of acetonitrile-1% phosphoric acid(15∶85, V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,and column temperature was 30 ℃. RESULTS:The lin-ear range of paeoniflorin was 0219-1.32 μg(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 95.82%-101.82%(RSD=2.13%,n=6). CONCLUSIONS:The method is simple,stable and reproducible, and can be used for the content determination of Bazhen pill(concentrated pill).

Objective To study the value of MSCT angiography(MSCTA)in diagnosing lower extremities arterial disease (LEAD) of diabetic foot ulcers and analyze the possible influential factors on the onset of diabetic foot ulcers.Methods 80 diabetic patients for MSCTA examination were divided into two groups by whether combined with foot ulcers.The tibial artery calcification score (TACS) and peripheral arterial occlusion index (PAOI) were calculated and the possible risk factors of diabetic foot ulcer were analyzed.Results There were 30 cases suffered with foot ulcer and 18 cases occurred lower extremity arterial occlusion (LEAO), the average PAOI was 4.40(3.60, 5.75).Non-foot ulcer cases were 50, and there were 8 cases suffered with LEAO, the average PAOI was 2.05(1.43, 3.10).Compared to patients without foot ulcer, patients with foot ulcer were older, had a lower BMI, and were more likely to have a history of tobacco use.They usually had higher TACS and severer peripheral arterial occlusive disease (PAOD).Stepwise regression model showed that foot ulcers had significant correlation with TACS and PAOI.Partial correlation analysis showed TACS was independent from PAOI in impacting diabetic foot ulcer.Conclusion MSCTA has important clinical value in diagnosing and valuing diabetic foot ulcer LEAD.Age, smoking history,BMI,TACS and PAOI affect the incidence of diabetic foot ulcers, and TACS and PAOI are the independent risk factors for diabetic foot ulcers.

Objective To investigate the role of β-arrestin2 in intestinal inflammation and illustrate the mechanisms from the perspective of epithelial barrier function. Methods Dextran sodium sulfate(DSS)is used to induce acute intestinal colitis in mice. The experiment groups are designed as the wild type control(WT),the wild type colitis (WT+DSS) and the β-arrestin2- knockout colitis (KO+DSS). The expression of β-arrestin2 gene by mRNA and protein level is compared between the WT and WT + DSS groups. The difference of weight loss , disease activity index(DAI),spleen weight,colon length,histological score,intestinal permeability and important tight junction proteins (occludin ,claudin1 and ZO-1) were detected in the WT+DSS and KO+DSS groups. Results Compared with the WT group,the expression of β-arrestin2 was significantly higher in the colon of the WT+DSS group. Compared with the WT+DSS group,the KO+DSS group had less weight loss(P < 0.05),lower DAI(P<0.05),smaller spleen,longer colon and lower histological score(P=0.002). The KO+DSS group had a lower intestinal permeability(P = 0.009)and higher protein level of occludin and claudin1.There was no signifi-cant difference of ZO-1 in the two groups. Conclusion β-arrestin2 may promote mouse colitis through impairment of epithelial barrier function.

Limonin existed in citrus fruits has been shown to have anti-bacterial, anti-viral, anti-feedant, anti-nociceptive, anti-inflammatory activities and anti-carcinogenic activities. But the clinical use is limited by its low bioavailability. The aim of this study is to observe the absorption and secretion transport mechanisms of limonin in intestine which can pave the way for the further study and clinical use. The transport characteristics and mechanisms of limonin in rat were studied by in situ intestine perfusion and in vitro Caco-2 cells method. The intestinal absorption of limonin was probably via a facilitated diffusion pathway which was poor and without segment-selection. Verapamil and ketoconazole improved the absorption remarkably according to the result of in vitro Caco-2 cells study; however, probenecid had no significant effect on the absorption. The P-gp efflux and CYP3A4 metabolism were involved in the poor intestinal absorption and low bioavailability of limonin. The exploration of the intestinal absorption mechanism is crucial to the design of dosage form and clinical use of limonin.

OBJECTIVE:To evaluate the effect of psychological intervention based on pharmaceutical care on the communi-ty patients with essential hypertension. METHODS:172 patients with essential hypertension in a community health service certer in Nanjing Gulou area in 2013 were divided into control group(n=87)and intervention group(n=85)by simple random meth-od. Control group received conventional antihypertensive drugs and management,intervention group additionally received pharma-ceutical care and psychological intervention,it lasted 6 months. Blood pressure controlling,drug compliance,mastery of drug knowledge and health status in 2 groups before and after treatment were compared. RESULTS:Before treatment,there were no significant differences in the systolic blood pressure(SBP)and diastolic blood pressure(DBP)between 2 groups(P>0.05);af-ter treatment, SBP and DBP in 2 groups were lower than before,SBP and DBP in intervention group were lower than control group,the decrease degree of SBP and DBP was bigger than control group,the differences were statistically significant (P<0.01). 1 month after the beginning of intervention,there was no significant difference in the proportion of drug compliance be-tween 2 groups (P>0.05);1 month after the end of the intervention,the proportion of drug compliance in intervention group was higher than 1 month after the beginning of intervention and control group,the differences were statistically significant(P<0.05). Before treatment,there was no significant difference in mastery of drug knowledge between 2 groups (P>0.05);after treatment,the proportion of patients showed good and excellent mastery of drug knowledge was higher than before and control group,the differences were statistically significant (P<0.05). Before treatment,there were no significant differences in scores of physical,mental,social health subscale and SRHMS between 2 groups(P>0.05);after treatment,scores of physical health subscale and SRHMS in control group,each subscale and to-tal scale scores in intervention group were higher than be-fore, and scores of mental, social health subscale and SRHMS in intervention group were higher than control group, the differences were statistically significant (P<0.05). CONCLUSIONS: The psychological intervention based on pharmaceutical care can effectively promote the con-trol of blood pressure,improve drug compliance,mastery of drug knowledge,health status and mental situation,which is worth of popularizing and applying in community health services.