Objective To investigate the relationship between neonatal health and TCM constitution types of early to mid-pregnancy expectant parents;To provide basis for the targeted interventions for couples of childbearing age before pregnancy-periconceptional TCM constitution. Methods Compiling scales for pregnant couple’ TCM constitution type assessment and postpartum visits to newborns, the research team got access to the information of neonatal physical problems, constitutions and changes of 675 babies’ father and mother who had established maternal manual from April, 2012 to March, 2013 at the Sanlin Community Health Center of Pudong District of Shanghai. Results Early to mid-pregnancy expectant parents mainly had peace constitution type. In detail, 55% of pregnant women had gentle constitution type, while others mainly had qi deficiency, yang deficiency and damp-heat constitution types;76% expectant fathers got peace constitution, while others mainly got damp-heat, yin deficiency and qi deficiency constitution types. Marital pregnancy physique did not change significantly. Compared with the peace constitution type, newborns, whose fathers got damp-heat constitution type, had higher risk of jaundice, and whose mothers got yin deficiency, the babies had higher risk of spits (P<0.05). The constitution types of expectant parents did not have obvious combined action on neonatal health. Conclusion There is a certain relationship between neonatal health and TCM constitution types of early to mid-pregnancy expectant parents. It is necessary to distinguish constitution types of prospective parents, and thus to take targeted health interventions.

Objective To investigate relationship between CD133 expression and progression of hepatocellular carcinoma (HCC) by detecting expressions of CD133. Methods 65 cases with HCC were enrolled and their HCC tissues and peripheral blood samples were taken. Expression of CD133 in HCC tissue was detected with immunohistochemical method and the percentages of CD45- CD133+ cells in peripheral blood were assayed by the method of flow cytometry (FCM), and 10 healthy persons were chosen as controls. Results In HCC patients, the positive expression rate of CD133 in HCC tissues was 6.4 % and the percentage of CD45- CD133+ cells in peripheral blood was 2.0 %. The expression rate of CD133 was not found in normal liver tissues and peripheral blood (P <0.01). The amount of CD133+ cells was significantly related to portal vein tumor embolus (P<0.01), tumor amicula integrality (P<0.01), pathology grade (P<0.01) and TNM stage (P <0.01), but not related to tumor diameter(P0.05) and AFP value (P0.05). It showed a positive correlation between HCC tissues and peripheral blood (r=0.36, P <0.01). Conclusion The percentage of CD133+ cell in HCC tissues and peripheral blood from HCC patient is greatly correlated to the progression of HCC. It is useful to exam the CD133 expression in predicting the diagnosis and prognosis evaluation.

Objective To study the effect of IL-8 on cell proliferation and invasion,and to analyze the correlations between chemokine and epidermal growth factor receptor(EGFR)signaling pathways in breast carcinoma cells.Methods IL-8 secretion responded to treatment with rhEGF and anti-EGFR and expression of its receptors CXCR1,CXCR2 in MDA-231 cells were measured by ELISA and immunocytochemistry,respectively.Effect of rhIL-8 and neutralizing antibody on cell proliferation and invasion were analyzed by using MTT and matrigel invasion assay.EGFR transactivation stimulated with rhIL-8 and neutralizing an tibody was assessed by Western blot using anti-phosphotyrosine antibody.Results MDA-231 cells released hish level of IL-8 and two receptom of IL-8(CXCR1 and CXCR2)both expressed on cell membrane.Exogenous IL-8 and its neutralizing antibody did not significantly influence the proliferation of breast carcinoma cells,but rhIL-8 stimulated invasive activity in MDA-231 cells and its neutralizing antibody inhibited the in vasive activity(P＜0.05).EGF and anti-EGFR both inhibited the secretion of IL-8 in breast carcinoma cells,and IL-8 had no effect on EGFR phosphorylation,but anti-IL-8 induced transactivation of EGFR after 24h.Conclusion IL-8 contributes to tumor progression in breast carcinoma through its enhancement of in vasive activitv but not act as an autocrine growth factor.The correlation of competitive inhibition rather than cross-talk is found between G protein coupled receptor(GPCR)-mediated IL-8 signaling pathway and EGFR pathway in breast carcinoma.

OBJECTIVE: To optimize the formula and preparation technology of gel-matrix sustained release tablet of nicotinic acid(GSTNA).METHODS: The formula of GSTNA was optimized by orthogonal experiment with the amount of hydrophilic gel-matrix material HPMC(K15M,E15-LV) and that of adjuvant calcium hydrogen phosphate(CHP) as factors and with the in vitro release rates as index.Meanwhile,the verification test on the intra-and inter-batch release rates of the samples was performed.RESULTS: The optimum formula could be seen as follows: the ratios of HPMC(K15M,E15-LV) and CHP were 4%,40% and 25% respectively.The GSTNA prepared in this formula achieved a sustained drug release of up to 12 h,and both the intra-batch homogenicity and the inter-batch reproducibility were satisfactory.CONCLUSION: The GSTNA is reasonable in formula and simple in preparation technology.

Objective To establish the triplex Taqman probes real-time RT-PCR method for simultaneously detecting of EV71,CA16 and EV.Methods Retrospective study.Specific primers and probes were designed based on conserved regions of EV71,CA16 and EV.The sensitivity,specificity and reproducibility were assessed by the optimized reaction system.A total of 176 throat swabs as the experimental group were collected from children with suspected hand foot mouth disease (HFMD),who admitted from April 2012 to July 2012 in Nanjing Children's Hospital affiliated to Nanjing Medical University.During this time,10 cases of healthy children,10 cases of outpatients with flu-like symptoms and 90 cases of inpatients in pneumology department of our hospital were recruited as control group,whose throat swabs were also collected.All of 286 samples were tested by the triplex Taqman probes real-time RT-PCR for simultaneously detecting EV71,CA16 and EV.SPSS13.0 was used to analyze the results.Results The sensitivities of the triplex Taqman probes real-time RT-PCR was 1.0 × 103 copies per milliliter for EV71,CA16 and EV.It showed 100％ specificity for 9 enterovirus and 3 non-enterovirus.Analysis with 1.0 × 103-1.0 × 105 copies per milliliter constructed plasmids demonstrated high reproducibility with coefficient of variation of 0.44％-1.04％ for EV71,0.38％-0.73％ for CA16,and 0.46％-0.90％ for EV.More over 176 samples collected from children with suspected HFMD were detected by triplex Taqman probes real-time RTPCR and real-time RT-PCR.The results showed 97.2％ (171/176)agreement and 0.94 Kappa value with high concordance.Conclusions The triplex Taqman probes real-time RT-PCR detecting EV71,CA16 and EV simultaneously has been established successfully.The assay,with high sensitivity and specificity,provide good basis for the rapid clinical diagnosis of EV71,CA16 and EV and open up broad prospects for clinical and relevant researches.

Objective To explore the relationship between peripheral Th17 cell number and chronic gastritis in Helicobacter pylori(H.pylori)-infected children.Methods Children were diagnosed as chronic gastritis by endoscopy.The degree and activity of inflammation were graded by histopathology examinations.The patients with both 13C urea breath test and urease test positive were diagnosed as H.pylori infection.The peripheral Th17 cell number was measured by flow cytometry and expressed as a ratio to total T cell.Results The Th17 cell number in HP group (chronic gastritis with H.pylori infection,n =33),non-HP group (chronic gastritis without H.pylori infection,n =24) and normal controls (n =15) were (1.55 ±0.30)％,(1.06 ±0.33)％,and (1.04 ±0.35)％,respectively.HP group included a statistically higher Th17 cell number than the other groups (all P ＜ 0.05),while no obvious difference was found between non-HP group and controls (P ＞ 0.05).According to the degree of inflammation,the chronic gastritis with H.pylori infection was categorized into non-apparent (n =10),mild (n =8),moderate (n =9) and severe (n =6) subgroups.The Th17 cell number in each subgroup was (1.64 ± 0.21)％ (non-apparent),(1.61 ± 0.23)％(mild),(1.25 ± 0.29) ％ (moderate) and (1.75 ± 0.20) ％ (severe),respectively.The moderate group had a lowest Th17 cell number among 4 groups (P ＜ 0.05).And significant differences did not exit in the other 3 groups (P ＞ 0.05).The HP group patients with different inflammatory activity had a Th17 cell number of (1.23 ±0.25)％ in nonapparent (n=15),(1.53 ±0.15)％ in mild (n=6),(1.55 ±0.32)％ in moderate (n=6) and (1.71 ±0.35)％ in severe (n =6) subgroup,respectively.However,there were no significant differences among 4 subgroups (P ＞ 0.05).Conclusions In the progress of chronic gastritis with H.pylori infection,Th17 cells may play a role as a double-edged sword by protecting and fighting against H.pylori infection and immunopathologic insults.This would provide more insights into the treatment of H.pylori infection.

Objective In order to identify cytotoxicity and specificity of the suicide gene systems which vascular endothelial growth factor receptor 2(KDR) and CMV were used as promoters respectively. Methods Two cell lines, HUVEC and CNE-2, one expresses KDR highly,another doesn't, were used as target cells for identifying the cytotoxicity and specificity of recombinant adenoviral vectors with herpes simplex virus thymidine kinase (HSV-tk) gene expression driven by KDR promoter and CMV promoter respectively (pAdKDR-tk and pAdCMV-tk) in Vitro by adding GCV. Results HUVEC and CNE-2 showed similarly sensitivity to GCV when they were transfected by pAdCMV-tk. However, They shown different sensitivity to GCV when they were transfected by pAdKDR-tk. When HUVEC was transfected in high MOI (MOI=100) and concentration of GCV was 50 ?g/ml, The cell growth rate decreased to (23.12?4.90)% .On the contrary, CNE-2 cells growth rate remained as high as (70.46 ?3.27)%. Conclusion Two kinds of cell line CNE-2 and HUVEC can be killed by pAdCMV-tk suicide gene system, but pAdKDR-tk suicide gene system only can kill HUVEC cell line. The result implied that pAdKDR-tk could kill the cells that express KDR specifitly.

Objective To discuss the effectiveness between arthroscope cleaning combined with exercise therapy and exercise therapy in senile osteoarthritis patients.Methods Two hundred and ten senile osteoarthritis patients were divided into operation group and non-operation group by random digits table method with 105 cases each.Patients in operation group received the arthroscope cleaning and exercise therapy,and patients in non-operation group received the exercise therapy alone.The visual analogue score (VAS) and international knee documentation committee knee evaluation form (IKDC) score before treatment and 1,6 and 12 months after treatment were compared.Results A total of 210 patients received the analysis of result without drop-out.There were no statistical differences in VAS and IKDC scores before treatment and 6 and 12 months after treatment between operation group and non-operation group (P ＞ 0.05).The VAS and IKDC scores 1,6,12 months after treatment were significantly improved compared with those before treatment in operation group,and there were statistical differences (P ＜ 0.05).The VAS and IKDC scores 6,12 months after treatment were significantly improved compared with those before treatment in non-operation group,and there were statistical differences (P ＜ 0.05).According to the IKDC score,in operation group 15 patients were excellent,61 patients were fine,19 patients were general and 10 patients were bad after treatment,and good effectiveness rate was 72.38％ (76/105).In non-operation group,14 patients were excellent,59 patients were fine,20 patients were general and 12 patients were bad after treatment,and good effectiveness rate was 69.52％ (73/105).There was no statistical difference in the good effectiveness rate between 2 groups (P ＞ 0.05).Conclusions Exercise therapy with guidance can significantly improve the function of senile knee joint osteoarthritis,and ease joint pain symptoms.Arthroscope cleaning combined with exercise therapy can relieve the joint acute inflammatory symptoms,but it has no significant clinical effectiveness in long-term treatment of osteoarthritis.

Objective:To get specific monoclonal antibodies ( mAbs) against PD-L1 which can block PD-1/PD-L1 binding, and explore the feasibility of its application on the treatment of chronic HBV infection preliminarily by in vitro and in vivo model. Methods:E. coli expression and series chromatography purification system were employed to get human and mouse PD-1/PD-L1 that had binding activity in vitro. By immunizing BALB/c mouse with purified recombination proteins of PD-L1,mAb hybridoma cell lines against PD-L1 were obtained. The reactivity with human/mice PD-L1 of individual antibody and the interaction blocking activity of the mAbs to PD-1/PD-L1 in vitro were examined by indirect chemiluminescence immune assay. Results: 8 cell lines against PD-L1 were obtained and 2 Anti-PDL1 mAbs (Ab5 &Ab6) performed strong immune activity to human/mice PD-L1 and blocking activity to PD-1/PD-L1. In the PBMC stimulation experiment of chronic HBV patient,Ab5 and Ab6 could promote theγ-IFN levels. With HBV in-fecting mice model,intravenous injections of these mAbs induced dramatically decrease of HBV DNA copies about 20 times, HBsAg levels in serum reduced to 30% of the baseline level. Conclusion:We obtained 2 PD-L1 mAbs with the reactivity to human/mice PD-L1 and blocking activity to PD-1/PD-L1. The 2 mAbs can promote T cell function in PBMC stimulation culture of chronic HBV patient, have significant antiviral effect in HBV transgenic mice and can be candidates for immunotherapy applications.

Objective To investigate the expressions of hepatorna stem cell surface marker CD133 CD90 in tissues of hepatocellular carcinoma (HCC) and evaluate their related clinical significances. Method The expressions of CD133 CD90 were detected by immunohistochemical method in HCC tissues of 93 patients, and normal liver tissues of 10 cases. Results Among 93 cases with HCC, the positive expression of CD133 were found in 71 cases (76.3%), and CD90 positive expression in 64 cases (68.8%), and the percentage of positive cells were (6.4±3.3)% and (4.3±3.9)% respectively. No positive expression of CD133 and CD90 was found in normal liver tissues (P<0.01). CD133, CD90 expressions in the HCC tissues of TNM stage Ⅲ-Ⅳ [(8.1±3.7)%,(5.7±4.2)%] were higher than those of TNM stage Ⅰ - Ⅱ [(4.1±2.3)%,(2.3±1.9)%] (P<0.01). Spearman correlation analysis indicated that the expressions of CD133 and CD90 were up-regulated as the pathohistology grades increased (P<0.05). Positive correlation was observed between CD133 and CD90 expression (r=0.402, P<0.01). Conclusions CD133, CD90 positive cells exist in HCC tissues, their expressions positively relate to the TNM stage and the pathohistology grades for HCC patients.