Abstract:Objective:To explore the involvement of signal transducer and activator of transcription 3(STAT3) in angiotensin Ⅱ(AngⅡ) induced vascular endothelial growth factor(VEGF) expression and the role of angiotensin converting enzyme(ACE) in regulating STAT3 phosphorylation in mesenchymal stem cells(MSCs). Methods:STAT3 phosphorylation under AngⅡ stimulation was analyzed by Western blot analysis. siRNA targeting STAT3 (STAT3 siRNA) was used to inhibit STAT3,and its effect on VEGF production was examined by measuring VEGF protein level in conditioned media and VEGF mRNA expression in MSCs with ELISA and real-time PCR,respectively. Besides,captopril was used to specifically inhibit ACE,and its effect on STAT3 phosphorylation was analyzed by Western blot. Results:① AngⅡ stimulation significantly increased STAT3 phosphorylation level;② Pretreatment with STAT3 siRNA markedly abolished the AngⅡ-induced increase of VEGF mRNA expression and protein synthesis;③ Captopril significantly inhibited the AngⅡ-induced STAT3 phosphorylation. Conclusion:STAT3 is involved in the AngⅡ-stimulated VEGF expression,and the AngⅡ-induced STAT3 phosphorylation is ACE dependent in MSCs.

Abstract:Objective:To explore the molecular regulatory mechanism of down-regulation of E-cadherin inducing GSK-3β phosphorylation deactivation in hepatocellular carcinoma (HCC). Methods:The stable cell line without E-cadherin expression was constructed by RNA interference method. The expression levels of E-cadherin,Akt,p-Akt,GSK-3β and p-GSK-3β were examined by Western blot analysis;The expression levels of p-GSK-3β in HepG2-E-cadherin-shRNA stable cell line were examined by Western blot analysis after 10 μmol/L LY294002 (PI3K inhibitors) treatment;The mRNA level of Egr-1 and PTEN in HepG2-E-cadherin-shRNA cells were tested by RT-PCR;The expression of PTEN was also examined by Western blot analysis. Results:Compared with the blank,E-cadherin expression level down-regulated 82.54%,and the phosphorylation level of GSK-3β and Akt upregulated 298.93% and 218.98%,respectively. With the stimulation of PI3K inhibitor (LY294002),the phosphorylation level of GSK-3β down-regulated to 48.13% when compared with the blank. The mRNA level of PTEN dropped to 76.14%,and Egr-1 to 66.89% when compared with the blank. The expression of PTEN protein also down-regulated to 53.77% when compared with the blank. Conclusion:Down-regulation of E-cadherin deactivated GSK-3β via PI3K/Akt signaling in HCC cells.

Abstract:Objective:To investigate the expression and cellular localization of Nemo-like kinase(NLK) in hippocampus of rat following cerebral ischemia reperfusion(IR). Methods:Sixty adult male Sprague-Dawley rats were randomly assigned to two groups:sham-operated group and IR group. According to different observation time after surgery,IR rats were individed into six subgroups(1 h,4 h,8 h,1 d,3 d and 7 d groups). A middle cerebral artery occlusion (MCAO) model in adult rats was constructed. The expression changes of NLK and active Caspase-3 in hippocampus of rat following cerebral IR were detected by Western blot. The cellular location and possible biological behavior of NLK was investigated by immunohistochemistry and immunofluorescence staining. Results:Western blot result revealed that NLK expression gradually increased,reached a peak at 8 h after cerebral IR,then decreased,and finally increased. There were significant differences in NLK expression between sham-operated group and IR group at 8 h after reperfusion (P < 0.05). Moreover,Caspase-3 expression progressively increased and reached a peak at the 3rd day,then decreased. There were significant differences in caspase-3 expression between these two groups at the 3rd day after reperfusion(P < 0.05). Immunofluorescence staining showed that NLK expression was principally located in neurons,especially pyramidal neurons of CA1 area in the hippocampus. Conclusion:The expression of NLK remarkably increased after cerebral IR,suggesting that NLK was involved in the development of cerebral ischemia in adult rats.

Abstract:Objective:To explore the effect of fluvastatin(Flu) on the expression of fibronectin(FN) in human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysate (HGPDS). Methods:After incubated in DMEM with 0.01%FBS for 24 h,cultured HPMCs were randomly divided into control group,HGPDS group,HGPDS+Flu group,Flu alone group and DMSO control group. HPMCs were treated for 3,6,12,24,36,48,72 h respectively and the cellular viability were detected by MTT colorimetry. The mRNA expression of FN was observed by RT-PCR. HPMCs were treated for 24 h and the levels of FN in the culture media were measured by ELISA. Results:The results of MTT revealed that lower viability was found in the cells treated with HGPDS and partialy restored in cells treated with different concentrations of Flu at 24 h and 36 h when compared with control group. The significant improvement of cell viability was shown in Flu treatment at concentration of 1 × 10-6 mol/L at the time of 24 h(P < 0.05). Compared with the normal control group,the mRNA and protein expressions of FN increased time-dependently in HPMCs treated with HGPDS(P < 0.05),which peaked at 6 h and 24 h respectively. The fluvastatin significantly inhibited the effects of HGPDS on the expression of FN in a dose-dependent manner(P < 0.05). Conclusion:HGPDS can increase FN expression in HPMCs,which will be inhibited by Flu.

Abstract:Objective:To investigate inhibitory effect of lactate dehydrogenase-A (LDHA) short hairpin RNA (shRNA) on the expression of LDHA in human cholangiocarcinoma cell line Hucct-1 and observe the effect of LDHA down-expression on the proliferation of Hucct-1 cells. Methods: Three pairs of oligonucleotides targeting LDHA were cloned into linearized pGPU6/GFP/Neo eukaryotic expression vector after annealing. Hucct-1 cells were transfected with the recombinant plasmids that were identified by sequencing. Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to evaluate the effect of RNA interference on the transcription of LDHA gene and the expression of LDHA protein respectively. Results: DNA sequencing confirmed that three recombinant plasmids were successfully constructed. The si-LDHA-1 and si-LDHA-3 showed higher interfering efficiency. Compared with that of negative control,the protein expression inhibition rates of si-LDHA-1 and si-LDHA-3 were (50 ± 3)% (P < 0.05) and(70 ± 2)%(P < 0.05). MTT assay showed that the proliferation of LDHA knocked-down Hucct1 cell (si-LDHA-3) was slower than negative control Hucct1 cell(si-NC),and there was significant difference in the absorbance value between transfected cells and negative control cells at 96 h after transfection(P < 0.05). Conclusion: The recombinant plasmid can effectively knockdown the expression of LDHA in Hucct1 cells,and two efficient interfering targets in LDHA gene are screened out. Inhibition of LDHA expression evidently results in inhibition of Hucct-1 proliferation.

Abstract:Objective:To investigate the expression of cyclin A2 in rat cardiac myocytes after acute myocardial infarction(AMI). Methods:The total 30 adult male SD rats were randomly divided into 6 groups averagely,sham-operated group and other 5 AMI groups,3d,1w,2w,3w and 4w after AMI. The expressions of cyclin A2 and phospho-histone H3(H3P) were detected by immunohistochemistry in each group. Results:Compared with sham-operated group,the number of cyclin A2 positive myocytes increased remarkably after AMI,but decreased 2 weeks later gradually. And after 4 weeks after AMI,there was no difference in cyclin A2 expression in heart tissue between AMI group and the sham-operated group. In each group H3P expression increased remarkably at 3d,1w,2w,3w after AMI in nucleus of some myocytes. The positive expression rate of H3P was 1.56% to 3.12% in heart tissue after AMI. Conclusion:The expression of cyclin A2 and H3P indicates that some adult cardiac myocytes can reenter cell cycle after AMI.

Abstract:Objective:To investigate the effect of human insulin and insulin analogues combined with or without metformin on the proliferation of human mammary carcinoma cell line MCF-7. Methods:Firstly,human mammary carcinoma cell line(MCF-7) was treated with different concentrations(1,10,100,1 000 nmol/L) of insulin analogues or human insulin. Secondly,MCF-7 cells was treated with insulin analogues or human insulin at the concentration of 1 000 nmol/L,and the cell proliferation was tested by CCK-8 assay after 24 h,48 h and 72 h treatment. Thirdly,MCF-7 cells were treated with different concentrations(0,2.5,5,10,20 mmol/L) of metformin combined with human insulin or insulin analogues(1 000 nmol/L each). The proliferation of MCF-7 cells was also examined by CCK-8 assay. Results:Both insulin analogues and human insulin could increase the proliferation of cell line MCF-7. Glargine,Lispro and human insulin had a dose-and time-dependent effect on the cell proliferation while Detemir and Aspart had only a time-dependent effect. Metfomin could inhibit the proliferation of MCF-7 cells in a dose-dependent response. Metformin could decrease the effect on the proliferation that was induced by human insulin and insulin analogues. Conclusion:Human insulin and insulin analogues couid increase the proliferation of human mammary carcinoma cell line MCF-7. Metformin could decrease the proliferation of MCF-7 induced by human insulin and insulin analogues.

Abstract:Objective:To illustrate the influence of ＂autoantibodies＂ in a post-partum period of a Graves’ disease animal model.Methods:Female Balb/c mice aged 6 weeks were randomly divided into experimental group and control group,and injected intramuscularly with Ad-TSHR-289(3.3 × 108 pfu/50 μl PBS) and control adenovirus(Ad-β-gal 3.3 × 108 pfu/50 μl PBS) respectively,three times at 3 weeks intervals.When the immunization was over,both groups were mated with male mice after getting a bit of serum from venae angularis for measurement of TRAb titer and T4 level. The mice were sacrificed 1 day after delivery,blood and thyroids were obtained. T4 level was detected by radio immunoassay. Levels of TRAb,TPOAb,TGAb were detected by electrochemiluminescence immunoassay. The thyroids were used for histological examination. Results:TRAb positive rates were 96.7% in experiment group(P < 0.05). The incidence of hyperthyroidism in mice after delivery was 73.3%,which was higher than that after the 3rd immunization (56.7%). TPOAb positive rates were 16.7% in experiment group. The titre of TGAb in experiment group increased siginificantly higher than that in control group(P < 0.05). Conclusion:An animal model of Graves’ disease is successfully prepared by immunization with Ad-TSHR-289. The TRAb maybe involved in the increase of TPOAb and TGAb in post-partum mice with Graves’ disease.

Abstract:Objective:To investigate the effects of BVT.2733 on the expression of Visfatin and insulin resistance in the diet-induced obese(DIO) mice model. Methods:The C57BL/6J mice were randomly divided into the normal diet group and high-fat diet (HFD) group. After 20 weeks,the obese mice were randomly divided into obese control group and BVT.2733 treatment group. And they were orally administered placebo and BVT.2733,respectively,for two weeks. The levels of plasma glucose and serum insulin were measured by biochemistry technology and radioimmunity. The visfatin mRNA expression in adipose tissue were analyzed by real-time quantitative PCR. And the morphology changes of adipocyte were also observed. Results:Comparing to normal controls,the weight,plasma glucose and serum insulin levels increased in HFD group(P < 0.05),and the size of adipocyte in HFD group was big. Comparing to obese controls,in BVT.2733 treatment group,the serum insulin significantly decreased(P < 0.01) and the adipocyte sizes reduced,while the mRNA expression of Visfatin down-regulated(P < 0.05). Conclusion:BVT.2733 could reduce body weight significantly,ameliorate insulin resistance,and down-regulate the expression of Visfatin.

Abstract:Objective:To explore the clinical value of chorionic villus cell cytogenetic analysis in early spontaneous abortion between natural and assisted reproductive technology(ART) pregnancy. Methods:Total 108 chorionic villus specimens that were obtained from 107 cases of early spontaneous abortion were cultured for karyotype analysis. Chorionic villus was successfully cultured in 106 cases,with 46 cases of natural pregnancy group(N group) and 60 cases of ART group,in which there were 7 cases of intrauterine insemination and 53 cases of in vitro fertilization coupled with intracytoplasmic sperm injection. Results:①The success culture rate were 98.15%(106/108). ②The normal karyotypes were identified in 28 cases of N group and 41 cases of ART group. The abnormal karyotypes were identified in 18 cases of N group and 19 cases of ART group. The abnormal karyotypes detection rate were 34.91%(37/106) in all cases. The type of numerical abnormalities was counting for 33.96%(36/106) and the type of structural abnormalities accounting for 0.94%(1/106). ③The majority type of abnormal karyotypes was numerical abnormalities,counting for 97.3%(36/37),and trisomies for chromosomes were common,counting for 75.68%(28/37). In N group,there was 1 case of triploids,1 case of sexual chromosome number abnomalities,and 16 cases of autosomal abnormality. In ART group,there were 2 cases of triploids,2 cases of tetraploids,1 case of robertsonian translocation,2 cases of sexual chromosome number abnomalities,and 12 cases of autosomal abnormality. ④ Males embryo were found 47.17%(50/106),females embryo were found 52.83%(56/106),there was no significant difference in sexuality ratio;In chromosome abnormalities,males embryo were found 48.65%(18/37),females embryo were found 51.35%(19/37),there was no significant difference in sexuality ratio of chromosome abnormality. ⑤Mean ages of patients were (29.61 ± 3.60)years old in N group and (31.98 ± 4.38)years old in ART group,with significant differences between 2 groups. Conclusion:Chromosome abnormalities are important factors of early spontaneous abortion,and analysis of chorionic villus chromosome provides an important theoretical basis for clinical etiological diagnosis and next pregnancy treatment.

Abstract:Objective:To investigate the predictive value of intrinsic QRS duration(IQRSd) for left ventricular function deterioration in patients dependent on right ventricular apical(RVA) pacing. Methods:In this study,42 patients with third degree atrioventricular block who underwent pacemaker implantation were included. Twenty-two patients with absolute reduction value of left ventricular ejection fraction(ΔLVEF)≥5% after pacing (group with ΔLVEF≥5%) and 20 patients with ΔLVEF<5% after pacing(group with ΔLVEF<5%) were enrolled to investigate the possible reasons or predictors for the cardiac function deterioration. The examination results of 12-lead electrocardiography and echocardiography,and clinical conditions were recorded for every patient before the implantation and during the follow-up. The cumulative pacing rate of pateints were also obtained at each follow-up visit. Results:After 77.3 months of follow-up,left ventricular ejection fraction(LVEF) decreased significantly from (64.20 ± 6.30)% to (40.60 ± 10.00)%(P < 0.001) while left atrial diameter (LAD) and left ventricular end-diastolic dimension(LVEDD) enlarged significantly in group with ΔLVEF≥5%[LAD:(34.77 ± 6.42)mm vs. (41.00 ± 7.45)mm;LVEDD:(49.82 ± 4.86)mm vs. (55.59 ± 8.44)mm;P < 0.001)]. However,the average LVEF,LAD and LVEDD levels in group with ΔLVEF<5% after pacing were not statistically different from those before pacing[LVEF:(65.40 ± 3.25)% vs. (64.94 ± 3.00)%;LAD:(37.40 ± 4.84)mm vs. (38.15 ± 5.83)mm;LVEDD:(48.30 ± 3.95)mm vs.(49.00 ± 3.87)mm]. Kaplan-Meier analysis revealed that LVEF diminution after pacing appeared significantly earlier in patients with IQRSd≥110 ms than that in patients with IQRS<110 ms. Chi Square Test showed the rate of LVEF diminution was significantly higher in patients with IQRSd≥110 ms than that in patients with IQRSd<110 ms(P < 0.05). COX regression analysis indicated that IQRSd≥110 ms was the independent predictor of left ventricular systolic function decrease (P < 0.05). Conclusion:Long-term RVA pacing leads to deterioration of left ventricular systolic function and cardiac remodeling. In patients with IQRSd≥110 ms,deterioration of left ventricular systolic function occured earlier and more serious than in those with IQRSd<110 ms. IQRSd≥110 ms is the independent predictor of LVEF reduction in patients dependent on right ventricular pacing.

Abstract:Objective:To establish a method of detecting cyclooxygenase (COX)-1,COX-2 and P-selectin in platelet-rich plasma (PRP) by flow cytometry(FCM),and to detect the expressions of COX-1,COX-2 and P-selectin in PRP from coronary artery disease (CAD) patients without using anti-platelet agents. Methods:Twenty CAD patients were recruited,who were going to receive coronary artery bypass graft(CABG) and had withdrawn aspirin and clopidogrel for at least 7 days. After overnight fasting,the patients’ venous blood was drawn and processed,then the PRP was incubated with fluorescence labeled antibody,and COX-1,COX-2 and P-selectin were detected by FCM. Results:The positive percent of COX-1,COX-2 in PRP of the enrolled 20 patients were (39.67 ± 9.20)% and (4.11 ± 2.74)%,respectively,and the positive percent of P-selectin in platelet was (13.56 ± 8.07)% after incubated with saline and(24.08 ± 13.49)% after incubated with arachidonic acid,the real positive percent of P-selectin in platelet was(10.27 ± 7.40)%. Conclusion:FCM can effectively detect the platelet expressions of COX-1,COX-2 and P-selectin. There would be ethnic differences between Chinese Han and Caucasian regarding the platelet expressions of COX-1,COX-2 and P-selectin in CAD patients.

Abstract:Objective:To evaluate the association of 25-hydroxyvitamin D3[25 (OH)D3] with macroangiopathy in type 2 diabetes mellitus (T2DM). Methods:There were 189 T2DM patients and 100 controls in study. The patients were divided into two groups (T2DM group and T2DM complicated with macroangiopathy group) based on their clinical features. Clinical data,metabolic indices and serum 25(OH)D3 levels were detected compared among all groups. According to the level of 25 (OH)D3,all patients were divided into 3 groups,low 25 (OH)D3 group,middle 25 (OH)D3 group and high 25 (OH)D3 group. The relevance of 25 (OH)D3 and other parameters were analyzed by Pearson correlation analysis. Results:The serum 25 (OH)D3 level in T2DM group or T2DM complicated with macroangiopathy group was significantly lower than that in the control group(P < 0.05),and the serum 25 (OH)D3 level in T2DM complicated with macroangiopathy group was significantly lower than that in the T2DM group (P < 0.05). From high 25(OH)D3 level group to low 25 (OH)D3 level group,the prevalence of macroangiopathy were increased from 14.29%,25.40% to 48.73%,respectively. The macroangiopathy prevalence in T2DM of low 25 (OH)D3 level group was significantly higher than the other two groups with high 25 (OH)D3 level(P < 0.05). Conclusion:The serum 25(OH)D3 level was negatively related with macroangiopathy in T2DM. The decreased serum levels of 25(OH)D3 maybe increase risk of macroangiopathy.

Abstract:Objective:To observe the effects of continuous sugar monitoring system(CGMS) association with insulin pump (continuous subcutaneous insulin infusion,CSII) to treat type 2 diabetic mellitus patients. Methods:CGMS or finger blood (capilary blood glucose) 8 times per day was used to monitor and guide the insulin pump to treat diabetic patients,and then the treatment effects of patients were compared between the finger blood(capilary blood glucose) group and CGMS group. Results:The time of blood glucose becoming normal in CGMS group was significantly shorter than finger blood group,sever hypoglycemia occurred less in CGMS group than in finger blood group. Conclusion:Using CGMS association with CSII to control blood glucose was safer and faster than using finger blood to monitoring blood glucose group.

Abstract:Objective:To investigate the relationship between serum levels of cystatin C(Cys C)and and venous leg ulceration(VLU). Methods:Subjects were divided into three groups,chronic venous insufficiency(CVI) patients without ulcers,patients with VLU and normal subjects. Enzyme-linked immunosorbent assay(ELISA) was used for detection of plasma TNF-α,IL-1β,IL-2R,Cys C levels and Cys C level of venous tissue. Results:① The plasma levels of TNF-α and IL-1β in the VLU patients with venous ulcers were(185.46 ± 20.90) pmol/L and(100.60 ± 17.43) pmol/L respectively,and was significantly higher than those in the control group [(32.28 ± 7.76) pmol/L of TNF-α and (58.72± 16.20) pmol/L of IL-1β](P < 0.05). The plasma levels of TNF-α and IL-1β in the CVI without ulcers group were(41.68 ± 7.35) pmol/L and(65.36 ± 25.51) pmol/L,and had no significant difference compared to the control group (P > 0.05);The plasma IL-2R levels in the VLU group and CVI without ulcers group were (138.56 ± 78.75) pmol/L and(94.45 ± 35.26) pmol/L,and both of them had no significant difference compared to the control group[(93.67 ± 12.28) pmol/L](P > 0.05);②Cys C expression in venous tissue was significantly lower in VLU patients than in the CVI without ulcers and control group(P < 0.05);The plasma Cys C level of VLU group was significantly lower than that of control before surgery (P < 0.05),and the plasma Cys C level of VLU group four days after surgery was significantly higher than that before surgery(P < 0.05). ③ The correlation analysis showed that Cys C expression was significantly negatively correlated with expressions of TNF-α and IL-1β(r = -0.878 and -0.990,P <0.05). ④After surgery,ulcer recurrence rate in patients with low expression of Cys C were higher than that with the high Cys C expression. Conclusion:Cys C expression is closely related to the incidence of VLU. Low expression of the Cys C may be an important factor of the incidence and recurrence of VLU.

Abstract:Objective:To evaluate effect of fresh aloe treating phlebitis and find a way to cure phlebitis. Methods:Thirty healthy large-eared New Zealand rabbits were chozed. Two edged-veins in both ears were set intravenous indwelling needle mimicing real clinic transfusion. The indwelling needles were extracted when phlebitis occurred 5 days after needles setting. Then they were randomly divided into model group,experimental group Ⅰ and experimental group Ⅱ,10 rabbits each. Model group did not give any treatment,and the specimens were taken immediately after group divided. The experimental group Ⅰ were given wet compress with 50% magnesium sulfate. Experimental group Ⅱ were given wet compress with fresh aloe, both groups given extra plastic wrap,4 h each time,2 times per day. After 5 days specimens were taken for pathological observation. Hematoxylin-eosin staining was used to observe pathological changes of vascular intima and beneath connective tissue,transmission electron microscope to observe ultrastructural changes of vascular endothelial cells. Results:There were significant statistical differences in inflammatory effect among three groups (P < 0.001);Transmission electron microscope showed that cytoplasm mitochondria and rough endoplasmic reticulum swelling in experimental group Ⅰ and Ⅱ was less than that in model group. And the swelling in experimental group Ⅱ was less than that in experimental group Ⅰ. Conclusion:Fresh aloe and 50% of magnesium sulfate are both useful to treat phlebitis,but fresh aloe treatment is more effective than 50% magnesium sulfate.

Abstract:Objective:To investigate the changes of inflammatory cytokines interleukin-8 (IL-8),tumor necrosis factor alpha(TNF-α)and high-sensitivity C-reactive protein(hs-CRP) in children in different phases of mycoplasma pneumoniae pneumonia(MPP). Methods:Sixty children with MPP and thirty healthy children were recruited from January 2009 to December 2011 in Nanjing Hospital Affiliated to NJMU. The mass concentration of the IL-8 and TNF-α was measured by enzyme linked immunosorbent assay(ELISA),the concentration of hs-CRP was measured by immune turbidimetry. The serum levels of the IL-8,TNF-α and hs-CRP were compared in different phases and stages with one-way analysis of variance(one-way ANOVA). Results:The serum levels of the IL-8 and TNF-α in the children with MPP in both acute stage and convalescence stage were significantly higher than normal control(P < 0.01). The levels of IL-8 and TNF-α in the severe cases were higher than those in the mild cases(P < 0.01). The level of the hs-CRP in the severe cases was higher than those in the mild cases and normal control in acute stage(P < 0.01). While there was no significant difference in the level of hs-CRP among three groups in convalescence stage (P > 0.05). Conclusion:Inflammatory cytokines of IL-8,TNF-α and hs-CRP play important roles in the pathogenesis of MPP. They can be used as monitoring indicators to reflect the severity of pneumonia.

Abstract:Objective:To investigate the protein and mRNA expression of monocarboxylate transporters 1(MCT1),CD147 and CD44 in non-small cell lung cancer(NSCLC),and significance of their clinical pathology. Methods:Fifty-six specimens of the NSCLC and peritumoral tissues and twenty-one control specimens of pulmonary benign diseases were studied. The mRNA and protein expressions of MCT1,CD147 and CD44 in specimens were analyzed by real-time quantitative reverse transcriptase polymerase chain reaction(real-time quantitative RT-PCR) and immunohistochemistry. Results:①The positive expression rates of MCT1,CD147 and CD44 were significantly higher in NSCLC tissues than those in the peritumoral and control tissues(P < 0.05);② The MCT1 expression in the cell membrane was correlated with both CD147 and CD44 expressions(P < 0.05);③ The expressions of MCT1,CD147 and CD44 in NSCLC were not correlated with age,gender,lesion dimension and TNM stage,but significantly correlated with histological types of tumor and lymph node metastasis (P < 0.05). And the mRNA and protein expressions of MCT1 and CD44 in NSCLC was correlated significantly with tumor differentiation degree (P < 0.05). Conclusion: MCT1,CD147 and CD44 may participate in the occurrence and development of NSCLC,and may be act as the diagnose and prognosis index of NSCLC.

Abstract:Objective:To investigate the expressions of thymidine kinase 1(TK1) and Ki-67 in non-small cell lung cancer(NSCLC)and their prognostic value in NSCLC. Methods:The tissue microarrays which contain 202 NSCLC cases and 63 normal lung cases were constructed. The expressions of TK1 and Ki-67 were detected in these tissues by immunohistochemistry method. The relationship between expression levels of two makers and the clinicopathological and survival parameters of NSCLC were evaluated. Results:The positive expression rates of TK1(60.89%)and Ki-67(55.45%)in the NSCLC tissues were significantly higher than those in normal tissue group (P < 0.000 1,P < 0.000 5). The Spearman correlation analysis showed that the expression of TK1 were positively correlated with Ki-67 expression (r = 0.518,P < 0.001). In the group of NSCLC, both TK1 and Ki-67 expression were significant correlated with tumor grade and TNM stage. Kaplan-Meier survival analysis indicated that the survival rate was markedly lower in patients with high expression level of two markers than those with low expression level (P < 0.05);Multivariate analysis using Cox regression model identified that TNM stage and TK1 expression were independent prognostic factors of NSCLC (P < 0.05). Conclusion:The expression of TK1 was significantly correlated with tumor grade and TNM stage. TK1 might be employed to evaluate the prognosis of NSCLC patients.

Abstract:Objective:To study the expression and clinical features of P53,Bcl-2,C-erbB-2 and (epidermal growth factor receptor,EGFR) in multiple primary bladder cancers and single bladder cancers,and provide evidence for diagnosis and targeted treatment of multiple primary bladder cancer. Methods:Immunohistochemical SP methods were used to detect the expression of P53,Bcl-2,C-erbB-2 and EGFR in 15 patients with multiple primary bladder cancers,15 patients with single bladder cancers(The clinical stage and pathological grade of patients with single bladder cancers were similar to those with multiple primary bladder cancers.) and 15 patients with normal bladder,the differences of the expression of P53,Bcl-2,C-erbB-2 and EGFR in each group were analyzed. Results:The positive expressions of P53,Bcl-2,C-erbB-2 and EGFR in patients with multiple primary bladder cancers and single bladder cancers were significantly higher than those in patients with normal bladders(P < 0.05). The positive expressions of P53 and Bcl-2 in multiple primary bladder cancer tissues were higher than those in single bladder cancer tissues(P < 0.05),while the positive expressions of C-erbB-2 and EGFR in multiple primary bladder cancer tissues were significantly lower than those in single bladder cancer tissues (P < 0.001). Conclusion:The positive expressions of P53,Bcl-2,C-erbB-2 and EGFR were different between multiple primary bladder cancers and single bladder cancers (P < 0.05). The combined detection of P53,bcl-2,C-erbB-2 and EGFR is valuable for the clinical diagnosis and identification of multiple primary bladder cancers. This study provides the possibility of the targeting therapy of multiple primary bladder cancers.

Abstract:Objective:To analyze expression of breast cancer suscepbility gene 2(BRCA2) in ovarian cancer tissues and its clinical significance. Methods:BRCA2 was detected using a pair of primer on 14 exon of BRCA2 by polymerase chain reaction amplification,and the mutations of BRCA2 were analyzed by DNA sequencing. The expression of BRCA2 protein in ovarian tumor tissues were detected by immunohistochemistry and Western blot. Results:No mutation was found in any mucinous cystadenoma and normal tissues,but the base T of 128 mutated C in 14 exon of BRCA2 in serous tumors and mucinous carcinoma. Western blot showed that BRCA2 protein expression of the malignant tumor in ovarian was lower than that of normal tissues and benign tumors. The results of immunohistochemistry showed that the BRCA2 expression rates in mucinous cystadenoma and mucinous carcinoma were 87.50% and 53.33% respectively,and there were significant differences between the mucinous cystadenoma and mucinous carcinoma (P < 0.05);whereas the BRCA2 expression rates in serous cystadenoma and serous carcinoma were 53.85% and 73.17% respectively,and there was no significant difference between the serous cystadenoma and serous carcinoma(P > 0.05). The BRCA2 expression was not significantly associated with patient’s age,clinical stage,differentiation and histological types of ovarian cancer (P > 0.05). Conclusion:BRCA2 expression in ovarian tumor tissues can be used as a diagnosis marker,it also may provide a target for the ovarian cancer treatment.

Abstract:Objective:To investigate the clinical significance of Fascin expression in esophagus carcinoma. Methods:Fascin protein expression was detected by immunohistochemistry(MaxVision method) using the monoclonal antibody and its intensity was assessed quantitatively by Image-pro plus image analysis system. Results:The expression of Fascin in the tissues of esophagus carcinoma was higher than that in normal esophagus tissues(P < 0.05),which correlated with clinical stage(P < 0.05),infiltration degree(P < 0.05),lymphatic metastasis(P < 0.01). The expression of Fascin was also associated with histological grade of esophagus squamous carcinoma. Conclusion:The higher expression of Fascin is the marker of high invasive and low histological grade of esophagus carcinoma,and Fascin may become a target in the therapy of esophagus carcinoma.

Abstract:Objective:To investigate whether ERβ gene expression and polymorphisms can modify the risk of gastric cancer. Methods:Total 100 cases of primary gastric cancer were enrolled as the test group,and 90 cases of gastric ulcer as the control group. Immunohistochemical SP method was used to detect ERβ protein,RT-PCR method to detect the polymorphisms of CA repeat sequence and G1082A in ERβ gene. Results:Positive expression rates of ERβ were 68.0% and 23.3% respectively in the test group and the control group,which had significant differences between two groups (P < 0.01). The positive ERβ expression rates of the subjects with low differentiation degree,high invasion depth,lymph node metastasis and five years death were higher than those with high differentiation degree,shallow invasion depth,without lymph node metastasis and five years survival. And the positive expression rate of ERβ in clinical stages ofⅠand Ⅱ were significantly lower than that in stage Ⅲ (P < 0.01,P < 0.05 respectively). ERβ expression had nothing to do with onset recurrence,age,tumor size,and histological types (P > 0.05). Genotype distribution accords with the Hardy-Weinberg (H-W) genetic balance in CA repeat sequence of ERβ of gastric cancer group and control group(P > 0.05);There was no siginifican difference in comparative genotype distribution of SS,SL,LL and allele frequency of S and L(P > 0.05). In G1082A polymorphism of ERβ,analysis result of RsaⅠand AluⅠdigestion showed that two groups were accord with the H-W genetic balance(P > 0.05);There was no significant difference in comparative genotype of rr,Rr,RR and genotype frequency of r,R/a,A(P > 0.05);but genotype frequency of aa in the test group was higher than that in the control group (P < 0.05). Genotype in G1082A polymorphism of ERβ was closely related to clinical stages,differentiation degree,infiltration depth,metastases situation and the five-year survival condition(P < 0.05),and it wasn’t related to onset recurrence,age,tumor size and histological types(P > 0.05). Only frequency of a/A was closely related to differentiation degree and infiltration depth of gastric cancer(P < 0.01). Conclusion:ERβ polymorphism has close correlation with clinical stages,differentiation degree,infiltration depth,metastases situation and the five-year survival condition in gastric cancer,and genotypes of aa,alleles of a/A in ERβ are critical factors in gastric cancer.

Abstract:Objective:To analyze HIV and sexual transmitted diseases related epidemiological characteristics on drug users in Jiangsu Province in 2011. Methods:According to the data of HIV surveillance on drug users in 2011,Excel database was established and statistical analysis of data was performed with SPSS 15.0 software. Results:Of 3 290 drug users,39.3%was traditional drug user,mainly heroin;54.0% was new type drug user,mainly methamphetamine. Methamphetamine users were more likely to be single,female,young and low-condom-use with commercial sex workers(27.34% has reported commercial sex,33.11% reported consistent condom use);prevalence of syphilis was 17.11% and 9.10% by ELISA and TRUST repeated tests respectively. Methamphetamine users have higher prevalence of syphilis than traditional drug users. Conclusion:The traditional drugs,mainly heroin users are relatively small in Jiangsu province. Methamphetamine as the representative of new drugs occupys the main status of new drug users. The infection rate of syphilis,unsafe sex behavior and risk of HIV spreading is higher in new type drug users than traditional drug users,and intensive intervention should be followed.

Abstract:Objective:To investigate the prevalence and risk factors for syphilis among men who have sex with men (MSM) in Nanjing. Methods:A face-face questionnaire interview was conducted among MSM by snowball sampling method in Nanjing from March to July,2009. Socio-demographic,behavioral and other characteristics were included in the questionnaire. Bivariate and multivariate logistic regression model was constructed to evaluate independent factor associated with syphilis. Results:A total of 661 MSM were recruited. The mean age of these participants was 27.44 years. Among them 84.0% were unmarried,and 90.5% had an education beyond senior middle school; in the past 6 months,5.0% had sexual contact with more than 10 male sex partners,the rate of protected anal sex during the past 6 months was 36.0%. The percentage of people who had ever paid for male sex service was 3.9%,who had provided commercial sex service to male was 4.5%,and who had sexual contact with female was 21.0%. Syphilis was diagnosed in 11.5% of these participants,the risk factors independently associated with syphilis clude being older than 25 years (adjusted OR=4.20,95%CI:2.26~7.80),non-local residency(adjusted OR=1.67,95%CI:1.00~2.78),looking for male sex partners from bathhouse(adjusted OR=2.29,95%CI:1.26~4.15),sexually transmitted diseases examination and treatment in 1 year before interview(adjusted OR=2.15,95%CI:1.25~3.69). Conclusion:The rate of unprotected anal sex was high, so was the prevalence of syphilis among MSM. Further efforts are urgently needed to scale-up the prevention of syphilis among MSM.

Abstract:Objective:To explore the differences of male infertility in different fluorine exposure populations. Methods:Villages were divided into high-fluorine-region group (n = 94) and low-fluorine-region group (n = 114). Their living situations,diet,fertility and other fluorine exposure related aspects were collected and compared between two groups. Results:Compared with low-fluorine-region group,it was significantly higher in the frequency of using coal and drinking tea,degree of dental fluorosis,and rate of infertility(P < 0.05),while significantly lower in the use of improved stoves and fluorine toothpaste in high-fluorine-region group(P < 0.05). The factors of “survey objects from high-fluorine region” and “stove unimprovement” significantly affected on male infertility in multiple Logistic regression(P < 0.05). Conclusion:Coal-burning-type fluorine exposure and the incidence rate of infertility in high-fluorine-region group was more serious than low-fluorine-region group,which suggested that long-term fluorine exposure was a risk factor of infertility,which could increase the risk of male infertility.

Abstract:Objective:To establish the expression profiles of peptides in sera of ovarian cancer and explore the early-diagnosis value of expression profiles of peptides in ovarian cancer. Methods:Magnetic binding-weak cation exchange beads (MB-WCX) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis were used to detect expression files of peptides. The early-diagnosis pattern was established by differential peptides peaks using the ClinProTool data analysis program. Results:In the mass range from 1 000 to 15 000 Da,8 peaks with clear differences in amplitude between the patients and health controls were detected,5 peaks with mass charge ratio(m/z) of 2 662.02, 2 653.68,5 859.21,5 901.69 and 5 937.06 were up-regulated notably,and 3 peaks with m/z of 1 857.82,2 074.28 and 2 856.33 were markedly down-regulated in patients with ovarian cancer. In specimens of early-stage ovarian cancer,the accuracy of characteristic proteomic patterns detection could be 81.8%(9/11),and the specificity showed 100%. Conclusion:MB-WCX separation coupled with MALDI-TOF-MS and ClinProTools analysis could be a desired method to find bio-markers in ovarian cancer,and could have a potential value in the early diagnosis of ovarian cancer.

Abstract:Objective:To detect urinary polypeptide patterns in stages of the type 2 diabetic nephropathy (DN) in order to screen and find out the potential biomarkers. Methods:A total of 68 urine samples from type 2 diabetic patients with normoalbuminuria(DM1,n = 24),microalbuminuria (DM2,n = 22)and macroalbuminuria (DM3,n = 22),and healthy controls (C,n = 20) were condensed by magnetic beads based weak cation exchange chromatography (MB-WCX),and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS). ClinProtTM software was used to profile and screen the polypeptide patterns in urine of the patients. Results:When the relative molecular mass < 12 000,there were significant differences in 15 protein peaks between group DM1 and group C(P < 0.01). There was one protein peak of significantly low expression in the group DM2,when compared to group C (P = 0.029 8). And there were significant differences in 10 protein peaks between group DM3 and group C (P < 0.01). Conclusion:MB-WCX coupled to MALDI-TOF-MS is a fast,convenient and high throughput analyzing method capable of screening some relatively specific and potential biomarkers from the urine of DN with different stages.

Abstract:Objective:To establish a simple method for determination of 7 phthalate esters (PEs) by gas chromatography/mass spectrometry (GC/MS). Methods:Vacuum cleaner was used for gathering the house dust,and n-hexane was added and extraction was performed in an ultrasonic bath. The supernatant was concentrated and then added with n-hexane to 0.2 ml. PEs were determined by GC/MS. Results:Seven PEs had highly efficient separation within 22 minutes and showed good linearity within 5~2 000 ng/ml range with correlation coefficients all beyond 0.998. The relative standard deviations were 5.66%~18.03%,and the recoveries were 87.73%~110.63%. Conclusion:This method was a economic,fast,simple,sensitive and reproducible enough for settled-dust PEs measurements in house dust with large sample size.