Abstract:Objective:Taking PDCD10 (homo sapiens programmed cell death 10) and MST4 (Mst3 and SOK1-related kinase) as drug targets, to establish a low molecular weight inhibitors screening method in vitro. Methods:The open reading frames of PDCD10 and MST4 were reconstructed into the prokaryotic expressive vector pGEX4T-2. The recombinant proteins were purified using GST affinity chromatograph, and then analyzed using Western blot. ELISA assay was utilized to detect the activity of purified proteins of GST-PDCD10 and GST-MST4 and screen for the inhibitor in 825 compounds, which were verified by the dual luciferase reporter gene assay in Elk1 signaling pathway. Results:Drug targets PDCD10 and MST4 inhibitors screening method was established successfully in vitro. One compound screened by this method was confirmed to inhibit the activity of PDCD10 and MST4. Conclusion: The method is of better parallelism and stability. The screened compound could also inhibit the activity of PDCD10 and MST4 in Elk1 signaling pathway.

Abstract:Objective: To evaluate the expression of Wnt/β-catenin and NF-κB pathway in hypopspadiac male rats treated by utero exposure to di-n-butyl phthalate(DBP). Methods:Twenty pregnant rats were randomly divided into two groups, and treated group was given DBP by gastric intubation at a dose of 800 mg/kg from gestation day(GD) 14 to GD18. On GD19,GT samples of hypospadiac male rats(from DBP treated group)and normal rats(from control group)were collected. Furthermore,Western blot analysis was used to quantify expression of β-catenin,GSK-3β and NF-κB in genital tubercle(GT) of hypospadiac male rats. Results:The body weight and the anogenital distance(AGD) of fetal male rats significantly decreased in DBP treated group. Moreover,the expression of β-catenin was significantly decreased, and an increased expression of GSK-3β and NF-κB in GT was observed(P < 0.05) in hypospadiac male rats. Conclusion:DBP could possibly affect the development of GT by regulating Wnt/β-catenin and NF-κB pathway in fetal male rats.

Abstract:Objective:To study the effects of polychlorinated biphenyls(PCBs) on the development of zebrafish embryo. Methods: We studied the influences of different concentrations (3.000, 2.000, 1.000, 0.500, 0.250, 0.125, 0.050 mg/L) of PCB1254 on morphological development, mortality rate and percentage of hatching using zebrafish as the model animal. Results: ①PCB1254 got teratogenic and mortality effect on zebrafish both at high and moderate concentration (above 0.25 mg/L). With the concentration increasing, hatching rate of zebrafish egg decreased while the mortality rate and the teratogenic rate risd obviously, which indicated that there was an obvious dose-depending effect; ②Deformities of zebrafish embryo exposed to PCB1254 mainly manifested spinal curvature and pericardialites; ③No effect was observed with the concentration of PCB1254 below 0.125 mg/L on the zebrafish; ④The 96 h LC50 of PCB1254 on the zebrafish was 1.082 mg/L, with 95% confidence interval ranging from 0.917 to 1.327 mg/L. Conclusion:Teratogenic and mortality effects are obersved on the development of zebrafish embryo exposed to different concentrations of PCB1254, which suggests the value of monitoring organic contaminant.

Abstract:Objective: To clone and express the human cytochrome P450 2E1(CYP2E1) gene. Methods: Using RT-PCR, a full length gene encoding CYP2E1 was successfully cloned and its expression was analyzed in the host cell E. coli BL21(DE3)by SDS-PAGE. Results: Except a C→T mutation at 1 263 nucleotide site, the nucleotide sequences of this CYP2E1 gene are consistent with the sequences in GenBank. The CYP2E1 gene was successfully inserted into the expression vector pET-32a (+) and expressed in the host cell E. coli BL21(DE3). The CYP2E1 protein with relative molecular mass of approximately 67ku was observed on the SDS-PAGE gel. Conclusion:The cloning and expression of CYP2E1 provide the foundation for following pre-clinical drug safety evaluation and the analysis of drug metabolism.

Abstract:Objective:To study the feasibility of selective extraction of chemical constituents from Polygonum cuspidatum with β-cyclodextrin. Methods:Chemical constituents of Polygonum cuspidatum were selectively extracted by crystallization of β-cyclodextrin inclusion complex in aqueous solution. Results:Six constituents in complex were shown by HPLC spectroscopy. Six compounds were isolated by semi-preparative reversed phase liquid chromatography and recrystallized. Five of the six compounds were identified:emodin, physcion, resveratrol, emodin-8-O-β-D-glucopyranoside and polydation. Conclusion:Chemical constituents of Polygonum cuspidatum could be selectively extracted with β-cyclodextrin. This method is a new way of extraction of chemical constituents from Chinese Herbal Medical.

Abstract:Objective:To prepare and characterize docetaxel-loaded nanoparticles(Doc-np) and to evaluate antitumor efficiency of Doc-np in vivo and in vitro. Methods: Doc-np was prepared by nano-precipitation method. Particle size was detected by dynamic light scattering,and the form was observed by atom force microscopy and transmission electron microscopy. Drug load content and encapsulation efficiency were measured by UV. In vitro release curve was also spotted. In vitro cytotoxicity was performed by MTT assay. Nude mice with transplanted tumor were used to measure the in vivo antitumor efficiency of Doc-np. Results: It was found in our study that Doc could be incorporated into the nanoparticles with high encapsulation efficiency more than 90%. In vitro release study showed that Doc was released from Doc-np in a sustained manner,and cytotoxicity studies indicated that IC50 of Doc-np against SKOV3 cells was significantly lower than that of free Doc. Furthermore, intratumoral administration was applied to improve the tumor-targeted delivery in the evaluation in vivo. Compared with free Doc, Doc-np exhibited superior antitumor effect by delaying tumor growth when delivered intratumorally. Conclusion: These results suggest that Doc-np are effective to inhibit the growth of human ovarian cancer, and intratumoral delivery of Doc-np could be a clinically useful therapeutic regimen and merit more research to evaluate the feasibility of clinical application.

Abstract:Objective: To research the effects of flavonoids of Astragalus complanatus(FAC) on proliferation,apoptosis and expressions of p53,c-My and NF-кB in human breast cancer MCF-7 cells. Methods:The inhibit rate of MCF-7 cells treated by different concentration of FAC was studied by MTT at different times. The apoptosis rate of MCF-7 cell was essayed by flow cytometry after treated with effective concentration of FAC and time points according to MTT. The expressions of p53, c-Myc and NF-кB were quantified by fluorescence immunoassay and Western blot. Results:According to MTT,0.0625,0.125,0,25,0.5 mg/ml of FAC treated for 24 h, 48 h,72 h markedly inhibited proliferation of MCF-7(P < 0.05,vs control). According to flow cytometry, MCF-7 was induced to apoptosis by FAC. After 72 h treatment with FAC(0.0625,0.125,0.25,0.5 mg/ml), the apoptosis rates were 6.5%,18.3%,49.6% and 57.9%,respectively. The expression of p53 was up-regulated, while c-Myc and NF-κB were markedly reduced detected by fluorescence immunoassay and Western blot. Conclusion:FAC could inhibit proliferation and induce apoptosis of MCF-7. Its role may be related to decreased expression of NF-кB and c-Myc, increased expression of p53.

Abstract:Objective: To observe whether reactive oxygen species(ROS) is involved in large dose ketamine-induced apoptosis on cultured primary neuron. Methods: Neuronal culture media (C group), 1 mmol/L ketamine (K group) or 1 mmol/L ketamine combined with 2.5 μmol/L superoxide dismutase mimetic M40403 (M group) was applied to primary neuron at day 6 for 24 h. Intraneuronal ROS production, the percentage of apoptotic cells and pro-apoptotic protein Bax expression were measured. Results: ROS production, the percentage of apoptotic cells and Bax expression in K group were 1.7, 4.2, and 2.0-fold of those in C group, respectively(all P < 0.05). Compared with C group, M group had no statistically significant changes in ROS production, the percentage of apoptotic cells and Bax expression (all P > 0.05). Conclusion: ROS could mediate the apoptosis induced by large dose ketamine in cultured primary neuron.

Abstract:Objective: To explore the effect of islet neogenesis-associated protein(INGAP) on islet function, apoptosis, gene expression and insulin secretion in vitro. Methods:Islets isolated from male adult Wistar rats were cultured in RPMI 1640 with or without INGAP(10 μg/ml). After 24 h, cultured islets were collected, the degree of apoptosis, glucose-stimulated insulin secretion were measured and the expression of genes Bcl-2 and Akt were detected by RT-PCR assay. Results: ① Glucose stimulated insulin secretion(GSIS) stimulated by 16.7 mmol/L glucose was increased in islets co-cultured with INGAP of compared with islets cultured without INGAP [(185.00±20.01) μU/ml vs.(58.67±17.03) μU/ml]; the stimulating index of islets of INGAP group(5.10±2.35) was higher than that of control (no INGAP group:1.92±0.42). ②Gene expression of Bcl-2 and Akt in INGAP group were increased compared with those of control group(0.607±0.217 vs. 0.503±0.209;1.115±0.189 vs. 0.935±0.162).③Islets survival in co-cultured pancreatic islet group(2 685.45±5.08)was higher than that of control group(2 108.65±4.02). Conclusion:INGAP can reduce the apoptosis of pancreatic islets and maintain islet function after co-culture in vitro.

Abstract:Objective: To define key genes relevant to insulin synthesis and exocytosis, and further to investigate the improved process of insulin secretion. Methods: Pancreata of SD rats at embryonic day 12.5(E12.5), E15.5, E18.5 and newborn, 21 days after birth, adult rats were dissected respectively. Genechips from Affymetrix company were applied to explore gene expression profiles. Some genes related to insulin synthesis and secretion were verified by real-time PCR. Intraperitoneal glucose tolerance tests (IPGTT) and ELISA were done to detect the function of pancreatic islet. Islet cells of newborn and adult rats were cultured to observe its shape. Results:Among genes related to insulin synthesis and release, insulin and relevant transcription factors such as PDX-1 began to express from E12.5; Munc13-1, Syntaxin-1a, Rab3a, Vamp-2, Glut-2 began to express from E15.5, while GCK expressed at late stage of embryonic day. Basic blood insulin level was detected at E18.5 and increased rapidly thereafter. However, in response to a glucose load, newborn rats were glucose intolerant,only rats older than a week began to show normal insulin secretory response gradually. The shape of islet cell in newborn rats was similar to adult one, but was much smaller and not so bright. Conclusion:Potential ability of insulin synthesis and release in rat occurs from early and middle stages of pancreatic development. But the normal insulin secretion response to glucose challenge starts later.

Abstract:Objective: To explore the relationship between human leucocyte antigen(HLA)-DRB1,DQB1 alleles and type 1 diabetes mellitus(T1DM) in Jiangsu area. Methods: We recruited 112 T1DM patients and 69 controls. The alleles were analyzed with polymerase chain reaction-sequence special oligonucleotide primers(PCR-SSO). The frequency of allele and haplotype were calculated by the chi-square test or Arlequin software. Results:17 DRB1 alleles and 7 DQB1 alleles were detected in 112 T1DM patients,the frenquencies of DRB1*0901, DRB1*0405, DRB1*0301, DQB1*0201 and DQB1*0303 alleles in the patients were significantly higher than that in healthy control group; the frequencies of DRB1*0901-DQB1*0303,DRB1*0301-DQB1*0201, DRB1*0405-DQB1*0401 and DRB1*0405-DQB1*0302 haplotypes were higher in T1DM than that in the controls. Conclusion: T1DM is associated with DRB1*0901, DRB1*0405, DRB1*0301, DQB1*0201 and DQB1*0303. And we find four susceptible DR-DQ haplotypes, DRB1*0901-DQB1*0303, DRB1*0301-DQB1*0201, DRB1*0405-DQB1*0401 and DRB1*0405-DQB1*0302 in Jiangsu area.

Abstract:Objective:To explore the relationship between islet autoantibodies of glutamic acid decarboxylase antibody (GADA), islet cell antibody(ICA) and insulin autoantibody(IAA) and HLA-A gene in patients with type 1 diabetes mellitus(T1DM) in Jiangsu province. Methods: One hundred and four patients with type 1 diabetes in Chinese Han of Jiangsu area and 110 healthy controls were recruited and measured for GADA, IAA, ICA by ELISA. HLA-A typing was performed by polymerase chain reaction-sequence specific oligonucleotide primer(PCR-SSO) technology after extracted genomic DNA from peripheral blood cells. The HLA-A alleles frequencies were studied by Arlequin software. Then an analysis was made on the correlation between islet autoantibodies of GADA, IAA, ICA and HLA-A alleles in type 1 diabetes mellitus by SPSS 11.0. Results: The positivity frequencies of GADA, ICA and IAA in 104 type 1 diabetes patients were 64.4%, 39.4% and 37.5%, respectively. The frenquencies of HLA-A*03 and HLA-A*24 alleles in the patients were significantly higher than that in healthy control group. The frenquencies of HLA-A*01, HLA-A*11, HLA-A*31 alleles in the patients of acute autoimmune diabetes were significantly lower than those in the healthy control group. In patients, whose onset age <20 years, the frequency of A*24 alleles was higher than that in the patients with onset age ≥20 years, but the frequency of A*31 alleles in acute-onset autoimmune diabetes with onset age <20 years was lower than that in the patients with onset age ≥20 years. The frequencies of GADA, ICA positivity were higher in those patients with A*03 alleles than those in patients lacking it; the frequency of ICA positivity was higher in those patients with A*24 alleles than those in patients lacking it. Conclusion: Our findings show that HLA-A*03 and A*24 are susceptibility to acute-onset autoimmune diabetes, whereas HLA-A*01, A*11 and A*31 are protective in Jiangsu province. A*31 may delay the onset of acute-onset autoimmune diabetes, A*31 is correlated positively with age at onset; while ICA, A*24 are negatively correlated with age at onset. A*3, A*24 are positively correlated with GADA, and A*24 is positively correlated with ICA.

Abstract:Objective: To explore a feasible method to detect tumor stem-like cells in human lung adenocarcinoma cells(LAC),and to analyze the properties of the sorted side population(SP)cells. Methods:Fluorescence-activated cell sorting assay(FACS)was performed to sort the SP and non-SP cells of LAC. MTT,colony formation and in vivo tumorigenesis assays were performed to analyze the proliferation capacity of SP and non-SP cells in vitro and in vivo. Flow cytometric assay was performed to detect the cell cycle and differentiation of SP cells. RT-PCR assay was performed to detect the expression of ATP-binding cassette transporter G2(ABCG2) mRNA in SP and non-SP cells. Results:Results from FACS showed that SP cells were detected in approximately 1.88% of LAC. MTT and in vitro colony formation assays showed that the in vitro proliferation capacity of SP cells was significantly stronger than that of non-SP cells. Additionally,the in vivo tumorigenesis capacity of SP cells was significantly stronger than that of non-SP cells. Cell cycle analysis indicated that the cell number of G0/G1 phase in SP cells was higher than that in non-SP cells(P < 0.05). The rates of SP cells in SP and non-SP populations were approximately (1.46±0.08)% and (0.12±0.04)%,respectively. Moreover,the level of ABCG2 mRNA in SP cells was 3.3 times of that in non-SP cells(P < 0.05). Conclusion:The SP cells sorted from LAC possessed characteristics of cancer stem-like cells,and SP cells sorted by FACS could be an effective method used to sort lung cancer stem cells.

Abstract:Objective: To induce allogeneic antigen-specific CD4+CD25+ regulatory T cells in vivo and identify the function of these cells. Methods: Allogeneic antigen-specific CD4+CD25+ regulatory T cells were generated by the addition of multiple intravenous injections of ICR mice splenocytes in vivo. Spleen cells were then pooled and enriched for CD4+CD25+ T cells and CD4+CD25- T cells by negative and positive selection using MACS in vitro. Characterizations of allogeneic antigen-specific CD4+CD25+ regulatory T cells were tested by mixed lymphocyte reaction(MLR) consisting of BALB/cByJ splenocytes as responder and MMC-treated ICR splenocytes as stimulator. The abilities of these cells to respond to various stimulators and to suppress alloreactive T cells responses were tested in vitro. Results: In the MLR, allogeneic antigen-specific T cells displayed little proliferation with allogeneic-antigen existence. Furthermore, these cells inhibited the proliferative responses and IFN-γ production of na-觙ve CD4+CD25- T cells to alloantigens in vitro. Conclusion: Donor-specific splenocyte transfusion-induced CD4+CD25+ T cells were allogeneic antigen-specific. They were anergic,and retained their ability to suppress antigen-driven responses of CD4+CD25- T cells in vitro.

Abstract:Objective:To investigate the effects of exogenous wild CSK-binding protein(CBP) gene stably transfection on growth and invastion of lung squamous carcinoma cells in vitro. Methods:A recombinant eukaryotic expression plasmid pcDNA3.0-CBP was constructed. Human lung cancer cell line NCI-H520 was transfected with pcDNA3.0-CBP or mock transfected plasmid pcDNA3.0(-)with lipofectamine2000, and cells stably expressing CBP were screened out by G418(800 mg/L).Changes of CBP protein and mRNA levels were measured by Western blotting and RT-PCR, cell viability was tested by MTT assay. Transwell and wound-healing methods were used to detect the difference of invasion and migration between transfected and non-transfected cells. Results: CBP protein and mRNA levels in pcDNA3.0-CBP transfected NCI-H520 cells were significantly higher than those in control NCI-H520 cells and pcDNA3.0(-) transfected NCI-H520 cells. The MTT assay revealed that the CBP gene transfected cells had less proliferative ability than control cells and pcDNA3.0(-) transfected NCI-H520 cells(both P < 0.01). As compared with control cells and pcDNA3.0(-) transfected NCI-H520 cells, the invasion and migration ability of pcDNA3.0-CBP transfected cells decreased obviously(P < 0.01). Conclusion: The CBP gene can restrain malignant phenotypes of the human lung squamous carcinoma in vitro and may participate in construction of negative feedback loop of SFKS to inhibit growth, migration and invasion of lung cancer cells.

Abstract:Objective: To investigate the effect of an antiangiogenesis therapy on inhibition of endometrium growth in the nude mouse model. Methods:Recombinant adenovirus was constructed with AdEasy-1 system and AAV 293 cells. Apoptosis of ECV-304 cells was induced by Ad-ES and growth inhibition was observed. The nude mouse endometriosis model was established by subcutaneous implantation. Injected in local focus with Ad-ES,Ad-Track or physiologic saline, the ectopic focuses were detected by microscopy after HE staining. The microvessel densities (MVD) were detected by immunohistochemistry and the apoptosis were detected by TUNEL. Results:The conduction of Ad-ES was proved by PCR, and titer of Ad-ES was 2.06×1010 pfu/ml. The apoptosis of ECV-304 cells induced by Ad-ES was analyzed by flow cytometer and Hoechst 33258 staining. The nude mouse endometriosis model was established by subcutaneous implantation, and the volume of endometriotic lesions treated by Ad-ES was deminished significantly compared with the other two control groups (P < 0.05). MVD and the apoptosis were decreased significantly in the group using Ad-ES compared with the two control groups. Conclusion:ES can induce ECV-304 cells to apoptosis and can inhibit the growth of endometrium in the nude mouse model. Antiangiogenesis may be a potential way for therapy of endometriosis.

Abstract:Objective: To characterize the effects of growth hormone (GH) on human colon cancer in vitro. Methods: Lovo cells were divided into control group and experimental groups and treated with different concentration of GH. MTT assay was used to determine the growth of the cells. The cell cycle and cell apoptosis of Lovo cells were detected by flow cytometer with PI and annexin V/PI double staining. Results: GH promoted the proliferation of Lovo cells, and the effects began at 12~24 h and peaked at 48 h. GH got the best effect on proliferation of Lovo cells when the concentration reached 200 ng/ml. After Lovo cells were treated by 200 ng/ml growth hormone for 24 h, the rate of G0/G1 stage decreased significantly, the rate of S stage increased significantly and the rate of G2/M stage increased slightly. For 72 h, the rate of G0/G1 stage decreased more significantly, the rate of S stage increased more significantly and the rate of G2/M stage increased significantly,the percentage of apoptotic cells also increased significantly. Conclusion: GH is a powerful promoter on proliferation of Lovo cells growth in culture. The effect of growth hormone manifesed a dose-dependent manner, and the saturation effect of growth hormone occurred at the concentration of 200 ng/ml, and its mechanism could be performed by promotion the cells G0/G1 stage to S and G2/M stage.

Abstract:Objective: To investigate the expression of prostate specific membrane antigen(PSMA) gene and protein in prostate cancer. Method:Expression of PSMA gene and protein were assayed by semiquantitive RT-PCR and immunohistochemistry method in 52 cases of prostate cancer and 35 cases of benign prostatic hyperplasia tissues. Result:①The positive rates of PSMA gene in the tissues of prostate cancer and benign prostatic hyperplasia were 84.6% and 68.6% respectively. Althought there was no significant difference between the two groups, the semiquantitive results showed that the expression level of PSMA gene in prostate cancer was significantly higher than in benign prostatic hyperplasia tissues. ②The positive expression rate of PSMA protein was significantly higher in prostate cancer tissue(94.2%) than that of in benign prostatic hyperplasia tissues(65.7%). Conclusion:The expression levels of PSMA gene and protein were higher in prostate cancer tissues than those of benign prostatic hyperplasia tissues, suggesting that PSMA may be a potential marker for prostate cancer.

Abstract:Objective: To study the expression and significance of C-erbB-2 and Ki67 in gastric carcinoma. Methods:The expression of C-erbB-2 and Ki67 were examined by SP immunohistochemical technique in 77 cases of primary gastric carcinoma samples. Results:The positive rates of C-erbB-2 in gastric carcinoma and gastritis cases were 62.3% and 6.7%, respectively, a significance existed between the two groups(P < 0.05). The expression rate of C-erbB-2 in gastric carcinoma was positively correlated with the invasion depth in gastric carcinoma(P < 0.05), and was not correlated with age, gender, differentiation and lymph node metastasis(P > 0.05). The positive rate of Ki67 in gastric carcinoma and gastritis cases were 77.9% and 10.0%, respectively, a significance was found between the two groups(P < 0.05). The positive rate of Ki67 in gastric carcinoma was not correlated with age, gender, invasion depth, differentiation and lymph node metastasis(P < 0.05). Conclusion:The over-expression of C-erbB-2 and Ki-67 may be involved in early stage of gastric carcinoma.

Abstract:Objective: To investigate the risk factors, clinical characteristics and coronary angiographic features in patients younger than 40 years with acute myocardial infarction(AMI). Methods: We collected data of patients with AMI from January 1999 to January 2009. Baseline clinical characteristics and coronary anatomy were compared between young (≤40 y) and older patients(60~75 y). Results: There were 106 cases in the young group and 112 cases in the older group. ST segment elevated myocardial infarction(STEMI) was presented in 91.5% patients of the young group, who were predominantly male,more likely to smoke, drinking, have higher coronary heart disease family history positive rate,and higher triglyceride(TG) levels,compared to older patients. Also,patients in the young group had less hypertension history and diabetic history, had lower pulse pressure and fasting blood glucose. The young group had higher rates of coronary angiography(64.2% vs. 40.2%,P < 0.05). Young patients were more likely to have single-vessel disease(58.82% vs. 13.33%,P < 0.05).Analysis of coronary angiogram also showed that the left anterior descending artery was the most commonly involved infarct related artery(56.63% vs. 32.29%,P < 0.01), no left main artery was involved in young group(0 vs. 10.42%, P < 0.01). Genisini scores were lower in young patients compared to the older group(23.69±22.80 vs. 39.21±40.37, P < 0.05). Multiple linear regression analysis showed that history of hypertension, Lp(a) and smoking index were independent influent factors for genisini scores in young patients with AMI. Conclusion:Smoking, drinking, family history and high TG were major risk factors for AMI in young who had less coronary pathological change, but strengthen the health education and improve the lifestyle such as hypertension control and smoking cessation should be strengthen to reduce the incidence of AMI in youngs.

Abstract:Objective: To explore the three dimensional distribution of the target sites of right ventricular outflow tract idiopathic ventricular arrhythmia(RVOT-PVC/VT). Methods:Sixty consecutive patients were included in the study. The target sites were identified as the earliest activation point by non-contact mapping, and were confirmed by successful ablation. The distribution of the target sites was observed in three dimensional geometry of RVOT. Results:Catheter ablation was successful in 49 patients whose target sites were located below the pulmonary valve. Among them,the target sites were located in the superior area in 15 patients, the inferior area in 34 patients, the anterior area in 30 patients, the posterior area in 19 patients, the free wall in 17 patients and the septum in 32 patients respectively. Catheter ablation was successful in 9 of 11 patients whose target sites were located above the pulmonary valve. Conclusion:Non-contact mapping could be helpful for identification the catheter ablation target sites of RVOT-PVC/VT.

Abstract:Objective: To estimate the therapeutic effect of intra-aortic balloon pump(IABP)on serious patients undergoing coronary artery bypass grafting. Methods:IAB catheters were inserted in the bodies of 31 serious patients undergoing coronary artery bypass grafting(LVEF≤40%) before operation and IABP worked after heart rebeating. Diastolic blood pressure,cardiac output,urine volume and dose of cardiovascular drugs were recorded before use and stopping of IABP. Results:Only one patient undergoing coronary artery bypass grafting died because of right heart insufficiency,and the total mortality was 3.2%.The other patients stayed in ICU for two to eight days. Diastolic blood pressure,cardiac output,urine volume before use of IABP were significantly lower than those before stopping of IABP. Dose of cardiovascular drugs before use of IABP was significantly higher than that before stopping of IABP. Conclusion:Application of IABP in the serious patients undergoing coronary artery bypass grafting can obstain good circulation support and elevate ratio of operation achievement.

Abstract:Objective: To investigate the endocrine and metabolic differences between polycystic ovary syndrome(PCOS) with nonalcoholic fatty liver disease(NAFLD) and without NAFLD. Methods: Body mass index(BMI), waist-to-hip ratio(WHR), blood pressure(BP), uric acid(UA), alanine transaminase(ALT), cortiso1(F), follicle stimulating hormone(FSH), prolactin (PRL), testosterone(T), estradiol(E2), luteinizing hormone(LH), total cholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C), lipoprotein a[LP(a)], fasting blood glucose(FBG), fasting serum insulin(FINS) and leptin(LEP) levels were measured in PCOS with NAFLD group(PCOS1 group,total number is 27) and PCOS without NAFLD(PCOS2 group,total number is 23). The HOMA-IR was measured too. Results: BMI,WHR,ALT,TG,LP(a),FINS,LEP were lower while HDL-C, F were higher in PCOS2 group than those in PCOS1 group(P < 0.05). HOMA-IR was positively correlated to LEP,BMI,Lp(a) in PCOS1 group, whereas HOMA-IR was positively correlated to T in PCOS2 group. Conclusion: Insulin resistance and dyslipidemia were more frequently observed in PCOS with NAFLD, while PCOS without NAFLD was demonstrated with higher T level. It is definitely that there are a clinical association between PCOS and NAFLD. The intervention and treatment of PCOS should be adopted differently between PCOS with NAFLD and PCOS without NAFLD patients.

Abstract:Objective: To observe average blood sugar,lipid profiles,serum uric acid level,cardio-cerebrovascular diseases and urinary albumin excretion incidence in type 2 diabetic patients with or without hypertension. Methods:Seven hundred and fifty five patients with type 2 diabetes were recruited and divided into normal blood pressure group(n=428)and hypertension group(n=324). Age,body mass index(BMI),HbA1c,total cholesterol(TC),triglyceride(TG),HDL-C,LDL-C,serum uric acid(SUA),urinary albumin excretion(UAE)rate were detected,the incidence of cardiovascular and cerebrovascular diseases were calculated. Results:Subjects with type 2 diabetes and primary hypertension accounted for 43.1% in this study population;age,gender,HbA1c and duration of diabetes in hypertension group were not different to those in normal blood pressure group(P > 0.05). BMI[(24.9±3.7) kg/m2],TC[(4.9±1.3) mmol/L],LDL-C[(3.1±1.2) mmol/L],TG[(2.5±3.0) mmol/L],SUA[(325.2±122.9) μmol/L] in hypertension group were higher than those in normal blood pressure group. The UAE rate[(47.7±91.4) mg/g] and positive ratio of UAE(24.3%),the cardiovascular and cerebrovascular diseases rate(15.1%)in hypertension group were higher significantly(P < 0.05,P < 0.01,respectively). Conclusion:Patients with type 2 diabetes mellitus and hypertension are prone to suffer from dyslipidemia,hyperuricemia,cadio-celebrovascular diseases and nephropathy than patients without hypertension.More attention should be paid to improve blood glucose,lipid profiles,uric acid and blood pressure control and prevent diabetic cardia-cerebrovascular and renal complications.

Abstract:Objective: To explore the value of quantitative analysis of contrast-enhanced ultrasonography for diagnosis and differential diagnosis of benign and malignant thyroid nodules. Methods: Sixty-four patients with thyroid nodules who had surgery in our hospital were selected to take contrast-enhanced ultrasonography,and then time-intensity curve(TIC) of contrast-enhanced ultrasonography was analyzed by computer software. Results:①These thyroid diseases which were diagnosed by conventional ultrasound and contrast-enhanced ultrasonography were highly consistent with the pathology results. ②The features of TIC: Most of papillary thyroid carcinomas were “quickly in and slowly out”, most of benign thyroid nodules (including nodular goiters and adenomas) were “simultaneously in and slowly out”. ③Features of quantitative parameters about TIC: The papillary thyroid carcinoma showed “more quickly in, the same intensity and more slowly out”; the nodular goiter showed “simultaneously in, lower intensity and more slowly out”; the adenoma showed “simultaneously in, the same intensity and more slowly out”. The speed of contrast excretion:The papillary carcinoma group was significantly slower than the adenoma group(P < 0.05); the papillary carcinoma group was significantly slower than the benign lesions(P < 0.01). Conclusion: Quantitative analysis of contrast-enhanced ultrasonography is useful in the diagnosis and differential diagnosis of papillary thyroid carcinoma and benign thyroid nodules, and has a good prospect.

Abstract:Objective:To investigate early diagnosis and treatment of tumor of duodenal papilla. Methods:The clinical data of 63 cases with tumor of duodenal papilla confirmed by operation and pathology from 1992 to 2009 were analyzed retrospectively. Results: The main clinical symptoms were jaundice(41 cases), digestive tract symptoms(18 cases) and hemorrhage of upper digestive tract (4 case). The diagnosis rate of FiScroptic duodcnocndoscopy, endoscopic retrograde cholangio pancreaticography (ERCP), magnetic resonance cholangiopancreatography(MRCP), type-B ultrasonic(BUS) and CT were 90.9%, 100%, 81.8%, 82.5% and 88.1%,respectively. Sixty-one cases underwent Whipple operation and two cases received local resection. Conclusion:ERCP and FiScroptic duodcnocndoscopy are the most effective methods for diagnosis of tumor of duodenal papilla. It is essential to early select radical resection operation so as to improve the result of surgical treatment.

Abstract:Objective: To evaluate the long-term effect of pancreaticoduodenectomy with no-touch isolation technique. Methods:Fifty-seven patients with ductal adenocarcinoma in the pancreatic head treated from 2004 to 2009 in Yancheng hospital were studied retrospectively. The patients were divided into two groups: NTIT(group A,n=32) and conventional techniques (group B,n=25). Blood samples(10 ml) were obtained through a catheter in the portal vein to detect CK20 mRNA expression by RT-PCR before and after surgery. The survival rate, liver metastasis rate and mortality were compared between the two groups. Results: As far as the mean volume of blood loss and the postoperative complications, there were no significant difference in the two groups(P=0.791,P > 0.05). Before the tumor resection, the overall positive rate of CK20 mRNA expression in portal vein blood was 17.54 %(10/54) , including 16.00% in conventional group and 18.75 % in NTIT group. This difference wasn＇t statistically significant(P=0.786,P > 0.05). After surgery, in 13 out of 25 cases (52.00%) of the conventional procedure group, CK20 mRNA was identified in the portal blood, while only 8 out of 32 cases (25.00%) was positive for portal CK20 mRNA in the NTIT group(P=0.024, P < 0.05). The survival rates of 1-, 3-, 5-year in group B were 76.6%,50.0%,40.0%, respectively, which were higher than those in group A. The rate of liver metastasis was significantly higher in group B than in group A(P=0.024, P < 0.05). Conclusion:In pancreatic cancer, the operation could cause the dissemination of cancer cells. The no-touch isolation technique may be a potential operative procedure to prevent cancer cells from being shed into the portal vein during surgical manipulation. NTIT could reduce the rate of liver metastasis so as to improve the long-term survival rate.

Abstract:Objective: To investigate the effects of dendritic cells(DCs) loaded with pancreatic cancer cells antigen on the immune function of the patients with unresectable pancreatic cancer. Methods:DCs isolated from peripheral blood mononuclear cell of pancreatic cancer patients were cultured and stimulated using rhGM-CSF and rhIL-4 in vitro,and then were pulsed with pancreatic cancer cells antigen. Thirty patients with unresectable pancreatic cancer were randomly divided into 2 groups. The dendritic cell vaccine was used in 15 cases(group A),and chemotherapy was applied in 15 cases(group B) as controls. Immune function and clinical effects were compared between the two groups. Results:The levels of CD3+-CD3+CD4+-CD4+/CD8+ and NK cell in group A were increased after vaccination(P < 0.05). So did the levels of IL-2,IL-12,IFN-γ(P < 0.05). Conclusion:The DCs vaccines loaded with pancreatic cancer cells antigen could improve the immune function of the patients with unresectable pancreatic cancer.

Abstract:Objective: To evaluate the efficacy and toxicity of paclitaxel liposome combined with 5-Fu/CF and DDP in treatment of patients with advanced gastric cancer. Methods:Fifty-six patients with advanced gastric cancer were divided into two groups randomly, the test group and the control group. The test group was treated with paclitaxel liposome 135 mg/m2 on day 1, 5-Fu 500 mg/m2 and CF 200 mg on day 1~5, DDP 20 mg/m2 on day 2~5. The control group was treated with paclitaxel 135 mg/m2 on day 1, 5-Fu 500 mg/m2 and CF 200mg on day 1~5, DDP 20 mg/m2 on day 2~5. The regimen was repeated every 21 days. All patients received at least for 2 cycles. Result: In the test group, the response rate(RR) and the disease controlled rate(DCR) were 46.4%, 78.5% respectively. One-year survival was 39.1%. In the control group, the RR, DCR and one-year survival were 32.1%, 50.0% and 17.8% respectively. The DCR in two groups had significant deviation. About adverse reactions, granulocytopenia, nausea, vomiting and allergy in the test group were decreased significantly compared with the control group. Conclusion: Paclitaxel liposome combined with 5-Fu, CF and DDP is active in patients with advanced gastric cancer and has less toxicity than paclitaxel.