Objective To construct a recombinant adenovirus contained respiratory syncytial virus F gene fragment. Methods The F gene fragment was amplified by RT-PCR. Then the recombinant shuttle plasmid pShuttle-CMV/F was constructed. The iinearized pShuttle-CMV/F was transformed into BJ5183-AD-1 electroporation competent cells which contained plasmid pAdEasy-1. The obtained homologous recom-binant plasmid was named pAdEasy/F, and transfected into 293 cells. Then the recombinant adenovirus rAd/F was obtained, and it was tested by electron microscope, RT-PCR, Western blot and immuno-fluores-cence assay(IFA). The vires titer and genetic stability were also studied initially. Results The recombi-nant adenovirus rAd/F was constructed, which was visualized as typical adenovirus morphology under elec-tron microscope. The transcription and expression of RSV F gene fragment integrated in the rAd/F were con-finned by RT-PCR, Western blot and IFA. Conclusion The recombinant adenovirus rAd/F contained RSV F gene fragment is successfully constructed and lay an important foundation for the further vaccine study.

Objective: To evaluate the applications and problems of statistical methods for articles published in《Chinese Circulation Journal》. Methods: We reviewed the statistical methods for original articles published in《Chinese Circulation Journal》in 2010, 2011 and 2015; general status with problems was analyzed and the situations were compared among different years. Results: A total of 404 articles were enrolled, statistical hypothesis testing was performed in 366 of them, 32 had descriptive statistics only and 6 articles were meta-analysis. The most frequently used statistical methods were student'st-test (57.4%), followed by chi-squared analysis (56.7%), analysis of variance (35.2%) and regression analysis (23.0%). The main statistical problems in Methods included in described method un-matched to really used method and insufifcient or wrong description. The proportion of insufifcient or wrong description in 2010 and 2011 was 36.6% (71/194), while in 2015 was 26.2% (55/210),P=0.02. The major wrong application of statistical approach was ignoring design with improper method and using unsatisifed precondition for speciifc analysis. The proportion of improper statistical method application in 2010 and 2011 was 19.1% (33/173), in 2015 was 19.7% (38/193),P=0.88. Conclusion: The quality of statistics was improved in 2015 than 2010 and 2011 for articles published in《Chinese Circulation Journal》; while there were still problems at certain degree, enhanced statistical review should be conducted in medical journals.

Objective To study the expression and clinical significance of thyroid transcription factor-1 (TTF-1) in stage Ⅰ non-small cell lung cancer (NSCLC) after operation and to 1 investigate the values in identification of the prognosis of stage Ⅰ NSCLC.Methods The expression of TTF-1 in 129 specimens of stage Ⅰ NSCLC was detected by immunohistochemistry.Results The positive rate of TTF-1 in stage Ⅰ NSCLC was 64.3％.There were significant differences in TTF-1 expression rate among pathological subtypes (x2 =25.231,P ＜ 0.001),different ages (x2 =4.581,P =0.032),sex (x2 =4.900,P =0.027) and differentiation degrees(x2 =11.519,P =0.019).Univariate analysis suggested that in the stage Ⅰ NSCLC patients,the median disease-free survival and overall survival of those positive for TTF-1 expression were superior to those negative:38.9 months versus 27.8 months (P =0.023) and 64.10 months versus 50.68months (P =0.013).Cox regression analysis suggested that TTF-1 expression,tumor differentiation degrees were independent factors affecting the prognosis of stage Ⅰ NSCLC.Conclusion Patients with TTF-1 positive expression often had better survival.Positive expression of TTF-1 and differentiation degree of tumor were independent prognostic factors to affect the prognosis of NSCLC.

Objective: To evaluate the intervention effect of clinical pharmacists in hypertension chronic disease management. Methods:All the patients with hypertensive chronic diseases from Ziyang community, Xingan Street, Beilun district were involved in the study. Combined with community doctors, clinical pharmacists provided pharmaceutical care for the patients, such as regular face-to-face medication guide, telephone communication, home follow-up, special lectures on health and so on. The cognitive level, blood pressure control level and medication compliance were statistically analyzed and compared before and after the pharmacy intervention. Results:After the intervention of clinical pharmacists, the level of hypertension cognition and the level of antihypertensive drug under-standingof the patients was improved significantly (P<0. 05 or 0. 01), the level of blood pressure control and medication compliance of the patients were improved significantly (P<0. 01), and unscheduled outpatient rate, emergency rate, hospitalization rate and fre-quency were decreased (P<0. 05 or 0. 01). Conclusion: Pharmacy intervention carried out by clinical pharmacists for the patients with hypertension chronic diseases can provide reasonable medication security and improve the quality of life, and the pharmacy inter-vention mode for the hypertension chronic disease management is worthy of promotion.

Objective To investigate the relationship between level of serum tumor necrosis factorα (TNF-α) and interleukin-12 (IL-12) and the antitumor effects of cantharidin sodium vitamin B6.Methods Forty advanced non-small cell lung cancer patients aged 30 to 80 were divided into experimental groups [paclitaxel + cisplatin (TP program) and sodium cantharidate vitamin B6] and control group (only TP program).Peripheral blood was drawn in the day of admission and the seventh day,the level of serum TNF-α and IL-12 were detected by method of enzyme-linked immunosorbent assay (ELISA),the results were analyzed.The World Health Organization (WHO) anticancer drugs in acute and subacute toxicity grading criteria were used to grade the chemotherapy side effects.Results The level of serum TNF-α and IL-12 were no significant difference on admission.After chemotherapy,serum TNF-α and IL-12 of experimental group was significantly higher than control group (P ＜ 0.000 and P ＜ 0.037,respectively).The serum TNF-α and IL-12 of experimental group had upward trend after chemotherapy.After 3 months of treatment,the remission rate was 85.7％ in experimental group,significantly higher than 57.8％ in the control group (P ＜ 0.05).White blood cells,and platelets decrease in the experimental group were significantly reduced than the control group (P ＜ 0.001 and P ＜ 0.009,respectively),and there are no significant difference in nausea and vomiting reaction between experimental and control groups.Conclusions The antitumor effects of cantharidin sodium vitamin B6 might be related to the high level of serum TNF-α and IL-12,cantharidin sodium vitamin B6 in combination with chemotherapy in patients can reduce white blood cell and platelet decline after chemotherapy.

Objective To evaluate the role of c-Jun N-terminal kinase (JNK) signaling pathway in paclitaxel-induced apoptosis in hippocampal neurons of rats, and the relationship with nuclear factor kappa B (NF-κB) pathway.Methods The primarily cultured hippocampal neurons were seeded in 96-well plate at a density of 1×106 cells/ml (200 μl/hole) , and were randomly divided into 4 groups (n=8 each) using a random number table: control group (C group), paclitaxel group (P group), JNK inhibitor SP600125 group (S group), and SP600125 + paclitaxel group (S+P group).Paclitaxel 2 ml (1 μmol/L) was added to group P.SP600125 2 ml (10 μmol/L) was added to group S.In group S+P, SP600125 2 ml (10 μmol/L) was added, the cell were then incubated for 1 h, and then paclitaxel 2 ml (1 μmol/L) was added.The cells were then incubated for 24 h.At 24 h of incubation, the apoptosis in hippocampal neurons was detected by flow cytometry, and the expression of NF-κB p65 was measured by Western blot.The apoptosis rate was calculated.Results Compared with group C, the apoptosis rate was significantly increased, and the expression of NF-κB p65 was up-regulated in P and S+P groups, and the apoptosis rate was significantly decreased, and the expression of NF-κB p65 was down-regulated in group S (P＜0.05).Compared with group P, the apoptosis rate was significantly decreased, and the expression of NF-κB p65 was down-regulated in group S+P (P＜0.05).Conclusion JNK signaling pathway mediates paclitaxel-induced apoptosis in hippocampal neurons of rats, and the mechanism is likely related to inhibition of NF-κB pathway activation.

Objective To investigate whether rat adipose-derived stem cells (rASCs) could resist human xenoantibody-dependent complement-mediated lysis and to explore its possible mechanisms.Method SD rat ASCs were isolated,rASCs at passage 2 to 8 were used for the following studies and rat lymphocytes were harvested as control cells.α-Gal expression was detected by flow cytometry.After incubation of rASCs with 20％ normal human serum (NHS) or heat inactivated normal human serum (HINHS),flow cytometry was used to detect cytotoxicity,IgG or IgM binding,and C3c,C4c and C5b-9 deposition.Result We successfully established the method to isolate and culture rASCs.The morphology of rASCs remained unchanged after passages.rASCs were positive for tell surface markers of CD44 and CD90,while negative for CD45 and MHC-Ⅱ.As compared with rLCs,rASCs significantly resisted human natural antibody and complement-mediated lysis when incubated with 20％ NHS in vitro (20.42％ ± 2.80％ vs 51.84％ ± 6.70％,P ＜ 0.01).Mechanistically,rASCs expressed lower level of α-Gal (13.97 ± 0.33 vs.24.47 ± 3.03,P＜0.05),which was correlated with decreased binding of human xenoreactive IgG and IgM (IgM:9.4 ± 2.0 vs.107.2± 4.8,P＜0.01; IgG:5.73 ± 1.0 vs.27.49 ± 3.9,P＜0.01) and reduced deposition of complements C3c,C4c and C5b-9 (C3c:294.6 ± 38.02 vs.1924 ± 509.4,P＜0.05; C4c:35.23 ± 3.1vs.177.3 ± 37.17,P＜0.05; C5b-9:5.63 ± 1.74 vs.37.05 ± 7.4,P＜0.01).Conclusion These data demonstrated that the resistance of rASCs to human xenoantibody and complement-mediated lysis is associated with low expression of xenoantigen a-Gal and inhibition of MAC (membrane attack complex) formation.

The serum TSH levels of 107 infants with subclinical hypothyroidism (SH) were > 20 mIU/L after 1-8 check-up, along with FT4, TT4, within low normal range. After given small dosage L-T4, for 4 weeks, blood TSH level obviously descended while FT4, TT4, ascended (all P <0.01). Seven cases of thyroid hypogenesis and 7 strumas were found by ultrasonography. It seems appropriate to use dosage of 3-4 μg·kg·-1·d-1 L-T4 in treating SH.

Positive linear correlation was found between TSH level and detection rate of congenital hypothyroidism (CH). When measured with a cut-off point at 9, 10, 15 and 20 mU/L of TSH, the sensitivity of positive detection rate for CH was 99.77%, 96.80%, 81.25% and 71.88%, respectively. TSH > 9 mU/L seems to be a reasonable cut-off value.

Objective To explore the differentiation of allogeneic naive T cells to regulatory T cells (Tregs) and T helper (TH) 1/2/17 cells by coculture with bone marrow-derivedimmature dendritic cells (irnDC).Method Bone marrow-derived imDC were cultivated from Balb/c mice.Lipopolysaccharide-stimulated DC were harvested as mature dendritic cells (mDC) and unstimulated cells were collected as imDC.Then irnDC or mDC were cocultured with allogeniec naive T cells,respectively.TH1 cytokines [interferon-γ (IFN-γ) and interleukin (IL)-2],TH2 cytokines (IL-4 and IL-10),and TH17 cytokine (IL-17) of co-cultured cells were detected by enzyme linked immunospot assay.CD4+ Forkhead box p3 (FoxP3) + Treg proportion in CD4+ cells in the co-cultured system with IL-2 and transforming growth factor-β1 (TGF-β1) was analyzed by flow cytometry.Result As compared with mDC,na(i)ve T cells cocultured with imDC secreted much less IFN-γ (11.67 ± 2.18 vs.182.00±23.71,P＜0.01),IL-2 (26.67±2.96 vs.318.30± 18.62,P＜0.01),IL-4 (17.00±3.78 vs.45.33±3.48,P＜0.01),IL-10(7.00±1.00vs.158.70±10.90,P＜0.001) and IL-17 (0.66 ± 0.33 vs.238.30 ± 24.39,P＜0.001).Furthermore,imDC induced more CD4+ FoxP3+ Tregs than mDC after adding IL-2 and TGF-β1 in the coculture system for 7 days (22.70 ± 1.53 ％ vs.5.42 ± 1.27％,P＜0.01).Conclusion imDC are more effective to induce na ve T cells to Tregs,but not differentiate to TH 1/TH 2/TH 17 cells.These findings provide in vitro experimental evidence for induction of transplant tolerance by adoptive transfer of imDC.