Objective To construct DNA vaccine expressing Mycobacterium tuberculosis(Mtb) immunodominant antigen Ag85A and analyze its anti-tuberculosis T cell responses in BCG primed-mice after DNA vaccination boosting.Methods The coding gene of Ag85A mature fragment was amplified by PCR with H37Rv genomic DNA as template,and then cloned into the eukaryotic expression vector pVAX1 to construct Ag85A DNA vaccine.After purification,Ag85A DNA vaccine was injected intramuscularly twice in BCG primed-mice with BCG vaccination and DNA vaccination alone as control.Eight weeks post-vaccination,spleen lymphocytes were separated and were then used to analyze Mtb antigen specific effector T cell response and polyfuntional IFN-γ/TNF-α/IL-2 secreting CD4+ T cell frequencies and intensities,and CD8+T cell responses by IFN-γ ELISPOT assay and intracellular staining,respectively.Results Compared to BCG vaccinated-and DNA vaccinated-mice,Ag85A DNA boosting not only enhanced significantly BCG primed-mice IFN-γ+TNF-α+IL-2+,IFN-γ+ IL-2+,TNF-α+IL-2+ and IL-2+ CD4+ T cell frequencies and IL-2 secretion,but also improved significantly IFN-γ-secreting and IL-2-secreting CD8+ T cell frequencies.Condusion Ag85A DNA vaccine was constructed successfully and was demonstrated to enhance significantly BCG primed-mice Mtb antigen specific CD4+ and CD8+ T cell responses when boosting,which is beneficial to improve BCG immunogenicity and its waning immune protection against Mtb.

Objective To investigate the expression of poly (ADP-ribose) polymerase-1 (PARP-1 ), cytokeratins (CK) 7/20, and p53 in patients with Barrett's esophagus and esophageal adenocareinoma,and to evaluate their significance. Methods Expression of PARP-1, CK7/20 and p53 were determined by immunohistochemistry in 108 patients (including 40 Barrett's esophagus, 28 esophageal adenocarcinoma and 40 cardiac mucosa). Results The expression of PARP-1 was found in Barrett's esophagus, esophageal ade-nocarcinoma and cardiac epithelium with a significantly higher level in esophageal adenocarcinoma than the other two groups (P <0. 01 ). CK7/20 was expressed in much of intestinal metaplasia, part of cardiac epi-thelium and adenocarcinoma cells. The positive expression of p53 was observed in all three groups, and it was significantly higher in adenocarcinoma group than in other two groups (P < 0. 05 ). PARP-1 expression is highly correlated with that of p53 in Barrett's esophagus ( r= 0.49, P < 0.01 ). Conclusion CK7/20 is a sensitive but less specific indicator for intestinal metaplasia. Both PARP-1 and p53 are involved in the patho-genesis of esophageal adenocarcinoma and might help to determine the risk of Barrett's esophagus developing into esophageal adenocarcinoma.

Objective To explore the mediating effect of cognitive emotion regulation strategies between emotional intelligence and loneliness. Methods 542 university students were surveyed by the Chinese version of Cognitive Emotion Regulation Questionnaire ( CERQ) ,the Emotional Intelligence Questionnaire and UCLA Loneli?ness Scale,and the correlation analysis was conducted by using SPSS17.0. The path model was set up and its path analysis was conducted by using AMOS17. 0. Results Positive cognitive emotion regulation strategies ( 28. 64 ± 5.90) were positively correlated with emotional intelligence(2.38±0.27, r=0.346, P<0.05) and negatively correla?ted with loneliness(42.29±7.88, r=0.301, P<0.05);Path analysis showed that the estimates of indirect and direct effects support the hypothesis respectively(P<0.05). Conclusion Positive cognitive emotion regulation strategies serve as partial mediators between emotional intelligence and loneliness.

Aim To investigate the effect of progester-one ( PROG) in protecting the neurons against impair-ment induced by the Aβ1-42 activated astrocytes, and the underlying molecular mechanism. Methods The astrocytes were divided into 5 groups: control, Aβ, and Aβplus PROG groups treated with 3 different con-centrations of progesterone for 24h. Then, Aβand pro-gesterone were removed, and neurons were co-cultured with the treated astrocytes. MTT assay was used to e-valuate the viability of cultured neurons; ELISA was used to detect the levels of IL-1βand TNF-αin culture media of astrocytes; immunofluorescence and Western blot were performed to detect the activation of NF-κB in astrocytes. Results PROG dose dependently pro-tected against Aβ1-42 activated astrocytes induced via-bility decrease in co-cultured neurons. Aβ induced release of IL-1β and TNF-α from astrocytes, and in-crease of NF-κB activity was abolished by progesterone treatment. Conclusion PROG protects the neurons through inhibiting the reactivity of astrocytes, and the underlying mechanism involves the NF-κB signal trans-duction.

Objective To evaluate the changes of left ventricular (LV) bulk rotation and untwisting in transplanted hearts using 2-dimensional speckle tracking imaging(STI).Methods Basal and apical LV short-axis images were acquired in 15 heart transplant recipients 3 months post surgery(HT group) and 56 healthy control subjects.Basal and apical rotation versus time profiles were drawn using 2-dimensional STI software.Appropriate values were chosen from the dataset obtained and compared between two groups.Results ①Compared with the control group,the heart rate,anterior-posterior diameter of left atrium,enddiastolic interventricular septum thickness,left ventricular posterior wall thickness,isovolumic relaxation time and E/e ratio were significantly increased,e and a values were decreased significantly in HT group (P ＜ 0.05).② No significant difference was noticed in the peak degrees of LV bulk rotation,the degrees of LV bulk rotation at the time of aortic valve closure and mitral valve opening (P =0.700,0.984,0.495,respectively) between 2 groups.In both groups,systolic rotation reached its maximum at end-systole [(96.1 ± 8.4) ％ in HT group vs (100.5 ± 6.3) ％ in control group,P =0.065].③Significant decreases in untwisting rate and trend untwisting variables were observed in the HT group(P ＜0.001).Conclusions 3 months after transplanted,left ventricular bulk rotation of cardiac allografts remained normal,and significant decreases in both untwisting rate and trend untwisting variables showed that the diastolic function of cardiac allografts was impaired.

Objective To investigate the causes and risk factors of postpartum hemorrhage (PPH) in hepatitis B virus (HBV) infected parturient.Methods Retrospective analysis was performed on the 1021 HBV infected parturient from Shanghai Public Health Clinical Center from July 2005 to June 2011.The comparisons were done by chi-square test.Results Among 1021 cases of HBV infected parturient,868 (85.01％) were asymptomatic and the PPH rate was 2.76％ (24/868) ;the remaining 153 cases (14.99％) were chronic active hepatitis B and the PPH rate was 16.99％(26/153).The difference between two groups was statistically significant (x2 =56.541,P＜0.01).The total incidence rate of PPH was 4.89％ (50/1021) and 17 cases (34.00％) were postpartum hemorrhage＞1000mL.The causes of PPH included uterine inertia (30/50,60.00％),abnormal placenta (11/50,22.00％),dysfunction of coagulation (5/50,10.00％) and lesion of birth canal (4/50,8.00％).The risk factors of PPH included delivery mode (x2 =6.528,P=0.038),abortion times (x2 =16.269,P=0.000),delivery times (x2 =6.990,P=0.008),ALT levels (x2=56.541,P=0.000) and HBV DNA (x2 =64.706,P=0.000).Conclusions The main causes of PPH in HBV infected parturient include uterine inertia,abnormal placenta,lesion of birth canal and dysfunction of blood coagulation.PPH is correlated with abortion times,delivery times,delivery mode,liver function and HBV DNA.The incidence of PPH in parturient with chronic active hepatitis B is higher than asymptomatic parturient.

Objective To assess the clinical significance of ankle-brachial index (ABI)in the diagnosis of peripheral arterial disease and its relationship with some interfering factors in type 2 diabetes mellitus. Methods One hundred and ten type 2 diabetes mellitus patients were selected.The systolic blood pressure and blood flow waveform in brachial artery,dorsal artery of foot or posterior tibial artery were surveyed with Doppler flow imaging survey meter ES-1000 SPM.ABI was calculated and the patients were divided into peripheral arterial disease group (group A) and non-peripheral arterial disease group (group B).Clinical data and risk factors were analyzed.Results In 110 patients,there were 20 cases (18.2%) with ABI<0.9.Age,TC,LDL-C,24 hours urinary microalbumin (24hUMA) and the prevalence of coronary heart disease were all higher in group A than those in group B (P＜0.05).LDL-C and the prevalence of coronary heart disease were independent risks variable for peripheral arterial disease. Conclusions ABI can assess objectively peripheral arterial disease in type 2 diabetes mellitus.which offers reliable base to active therapy.Advanced age,hypercholesteremia and hyper 24hUMA are the main factors of peripheral arterial disease.

This study was to investigate the permeability and absorbability of capsaicin cubosome across abdominal skin of the SD rats in vitro. Diffusion of capsaicin cubosome and cream was performed with the modified Franz diffusion cell technique. The capsaicin cubosome showed no enhancement of skin permeation within 24 hours. However, the deposition amounts of capsaicin in the rat skin in the cubosome group was markedly higher than those in the commercial cream group (P < 0.01). Cubosome showed excellent characetristic of skin-targed which could be a good carrier for the local transdermal drug delivery system.
Administration, Cutaneous ; Animals ; Capsaicin ; administration & dosage ; chemistry ; Kinetics ; Male ; Particle Size ; Permeability ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; metabolism ; Skin Absorption

OBJECTIVETo investigate the effects of different concentrations of lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), dexamethasone (Dex), and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro.

METHODSMCF7 breasr cancer cells were cultured and treated with different concentrations of LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L). Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot, respectively, after 6, 24, 48, and 72 hours of treatment.

RESULTSCompared with the blank con- trol, LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L) significantly increased the expressions of GPR54 mRNA (P < 0.05) and protein (P < 0.05).

CONCLUSIONLPS, TNFα, IL-6, Dex, and insulin evidently increase the expression of GPR54 in the MCF7 cell line, indicating their influence on the function of gonads by regulating the GPR54 level.

Blotting, Western ; Dexamethasone ; administration & dosage ; pharmacology ; Glucocorticoids ; administration & dosage ; pharmacology ; Gonads ; drug effects ; metabolism ; Humans ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Interleukin-6 ; administration & dosage ; pharmacology ; Lipopolysaccharides ; administration & dosage ; pharmacology ; MCF-7 Cells ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, G-Protein-Coupled ; drug effects ; genetics ; metabolism ; Receptors, Kisspeptin-1 ; Time Factors ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.