Abstract:objective:to observe the therapeutic effects of human fetal liver mesenchymal stem cells (hflmscs) induced by tat－pdx1 protein when transplanted into nod/scid diabetic mice. Methods:the hflmscs which were isolated and purified from fetal liver by adherent method,were induced to differentiate into islet β cells by tat－pdx1 protein. Nod/scid mice models of diabetes mellitus were established by injecting stz. The cells induced by tat－pdx1 protein were transplanted into left renal capsule,the blood glucose of diabetic nod/scid mice was detected. Results:the hflmscs induced by tat－pdx1 protein were expressed the mark of insulin in vitro. The level of mice blood glucose was degraded when the cells were transplanted into left renal capsule and was increased again when the left renal was excised. Conclusion:the hflmscs induced by tat－pdx1 protein can effectively reduce the level of nod/scid mice blood glucose. In the future,they will be the seed cells to treat diabetes mellitus.

Abstract:objective:to investigate the effects of exogenous wild csk－binding protein(cbp)gene stably transfection on growth of lung adencarcinoma cells in vitro. Methods:at first,a recombinant eukaryotic expression plasmid pcdna3.0－cbp was constructed. Human lung cancer cell line spc－a1 was transfected with pcdna3.0－cbp or mock transfected plasmid pcdna3.0(-)with lipofectamine 2000,and cells that can express cbp stably were screened out by g418(800 mg/l). Change of cbp protein level was measured by western－blot;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 level in pcdna3.0－cbp transfected spc－a1 cells was significantly higher than that in control spc－a1 cells and pcdna3.0 (-)transfected spc－a1 cells. The mtt assay revealed that the cbp gene transfected cells had less proliferation ability than control cells and pcdna3.0(-)transfected spc－a1 cells (p < 0.01). As compared with control cells and pcdna3.0(-)transfected spc－a1 cells,the invasion and migration ability of pcdna3.0－cbp transfected cells decreased obviously(p < 0.01). Conclusion:the cbp gene,as a recently identified transmembrane protein,can restrain malignant phenotypes of the human lung adencarcinoma in vitro and may participate in construction of negative feedback loop of sfks to inhibit lung cancer cells from growth,migration and invasion.

Abstract:objective:to compare the different features of t－cell－stimulation of dendritic－like cells (dc－like cells)induced from monocyte－macrophages derived from placentas in different gestational ages. Methods:monocyte－macrophages were separated mechanically from normal term placental tissues and second trimester placental tissues,then cultured in the transendothelial trafficking system and induced into dc－like cells. The dc－like cells were co－cultured with allogeneic t cells,in order to detect their abilities to stimulate allogeneic t cells into proliferation by cck－8 incorporation assay. Expressions of intracellular cytokines by stimulated t cells and the phenotype of dc－like cells were analyzed by facs. Results:after being cultured in the transendothelial trafficking system for 48 h,the cells which derived from the normal term placental monocyte－macrophages displayed characteristic morphology and expressed higher level surface antigen of cd80,cd86,hla－dr,and obviously stimulated the proliferation of allogeneic t cells,which could be led to a higher percentage of ifn－γ producing cells,but fewer of il－10 producing cells. However,dc－like cells,originated from second trimester placental tissues,expressed quite little levels of cd80,cd86 and hla－dr(p < 0.05)and stimulated very few allogeneic t cells to proliferation (p < 0.05). Furthermore,in the t cells stimulated by these dc－like cells ,there were small percentages of ifn－γ producing cells in cd8+ subset (p < 0.05)and large percentages of il－10 producing cells in both cd8- and cd8+ subsets. Conclusion:dc－like cells induced from monocyte－macrophages separated from different gestational ages placental tissues display functions in immunology with different biological characteristics.

Abstract:objective:to investigate the reversal effect of 4’－methylether－scutellarein(4’－m－s) on multidrug resistance in human choriocarcinoma jar/vp16 cell line. Methods:quantitative accumulation and efflux of rhodamine 123 (rh123),a p－glycoprotein(p－gp)substrate,were determined using a flow cytometric assay (fcm) for measuring p－gp function. Mdr1 mrna levels were detected by semi－quantitative reverse transcriptase－polymerase chain reaction (rt－pcr). The expressions of p－gp were analyzed by western blot and immunohistochemistry(ihc) method. Results:the results of fcm suggested that 20 μg/ml 4’－m－s could significantly increase the intracellular accumulation of rh123 in jar/vp16 cells. Rt－pcr and western blot results revealed that the expression levels of mdr1 mrna and p－gp protein were low in 4’－m－s treated cells. The ihc study showed that mild immunostaining for p－gp was seen in jar/vp16 cells when treated with 4’－m－s. Conclusion:the reversal effect of 4’－m－s in jar/vp16 cell may be related to regulating expression of mdr1/p－gp and increaseing intracellular concentration of chemotherapeutic drugs.

Abstract:objective:to investigate the cytotoxic effects of human peripheral blood γδt cells on lung cancer cell lines a549. methods:the γδt cells in peripheral blood of healthy volunteers were cultured using interleukin－2(il－2)and isopentenyl pyrophodphate (ipp)in prmi1640 medium. The purity of γδt cells was analyzed by flow cytometry .the cytotoxic activity of the purified γδt cells against human lung cancer cell lines a549 was tested in different effector－to－target ratios (t/e ratios). Results:γδt cells were obtained with a high purity from peripheral blood mononuclear cells,the cytotoxic effects of γδt cells on a549 depended on γδt cells’ purity and effector－to－target ratios. When effector－to－target ratios and purity of γδt cells increased,the cytotoxcity effect on a549 increased. Γδt cells showed more remarkable cytolysis than cik cells. Conclusion:γδt cells of human peripheral blood could significantly kill lung cancer cell lines a549 in vitro.

Abstract:objective:to explore the profiles of hepatoma cell lines－specific gene expression and identify the differential expressed cell apoptosis inhibitory genes for exploring hepatocarcinogenesis and new therapeutic targets. Methods:the profiles of gene expression of hepatoma cell lines bel－7402,hep－3b and fetal liver cells were analyzed by gene chips. The differential expressed cell apoptosis inhibitory genes were screened. The selected candidate genes were confirmed by rt－pcr in three kinds of cell lines and by immunohistochemistry in 20 specimens of hcc and adjacent noncancerous tissues. Results:of the detected 54 614 probe sets,compared with the fetal liver cells,1 452 genes were up－regulated by more than 2 folds and 2 054 genes were down－regulated by more than 2 folds in the hepatoma cell lines bel－7402 and hep－3b,in which 12 differential expressed cell apoptosis inhibitory genes were screened. The results of rt－pcr showed that the expressions of sodd and survivin in hepatoma cell lines were higher than those in fetal liver cells (p < 0.05). The positive rates of sodd and survivin were significantly higher in hcc than in adjacent noncancerous tissues(65% vs. 15%,and 75% vs. 10%,p < 0.05). Conclusion:many cell apoptosis inhibitory genes were up－regulated in hcc,which can inhibit cancer cell apoptosis and promote cancer cell proliferation by effecting tnf pathway,bcl－2 protein expression and so on. Analysis of these genes can provide important clues for disclosing the mechanisms of hepatocellular carcinoma pathogenesis and supplying possible new therapeutic targets.

Abstract:objective:to explore the significance of matrix metalloproteinase 2(mmp2) gene in colorectal cancer with liver metastasis. Methods:during operation,the fresh specimens of primary cancer lesions,surrounding normal colorectal mucosas,hepatic metastases and surrounding normal liver tissues were collected in 21 patients who suffered from colorectal cancer with liver metastasis. The mrna and protein expression of mmp2 among these specimens were detected by sybr green real－time rt－pcr and western blot method. Results:there were significant differences of mrna expression level in these specimens among 21 cases,which was highest in hepatic metastases and lowest in surrounding normal colorectal mucosas. The protein expression in most of cases (18/21) were lower in surrounding normal colorectal mucosas than other specimens by western blot. There was no difference between primary cancer lesions and hepatic metastases,and both of them were higher than surrounding normal liver tissues among all cases. Conclusion:the overexpression of mmp2 gene in primary cancer lesions and hepatic metastases were beneficial for degrading the extracellular matrix. It might be one of molecular mechanisms in colorectal cancer liver metastasis that the expression of mmp2 mrna in hepatic metastases was higher than that of primary cancer lesions due to the up－regulation of mmp2 gene. The overexpression of mmp2 in normal liver tissues surrounding hepatic metastases might be correlated with the regeneration of hepatic metastasis.

Abstract:objective:to investigate the effect of vagus nerve on first－phase insulin secretion. Methods:five healthy beagle dogs were anesthetized and cut off anterior vagal trunk and post vagal trunk of vagal celiac trunk. Intravenous glucose tolerance test and the continuous infusion of glucose via the carotid artery were performed in five beagle dogs prior to and after truncal vagotomy. Blood samples were collected from jugular vein of opposite side each time at the 0,1,2,4,6 and 10 min to measure plasma insulin and blood glucose levels. Results:during intravenous glucose tolerance test,first－phase insulin secretion obviously dropped after truncal vagotomy compared to prior(p < 0.05). But blood glucose failed to show significant difference before and after truncal vagotomy. The continuous infusion of low dose glucose via the carotid artery failed to induce any change in vein glucose level but caused an increasing of insulin secretion. It reached the peak at 2~4 min. Insulin secretion obviously dropped after truncal vagotomy(p < 0.05). Conclusion:the central nervous system may play a role in regulating first－phase insulin secretion,which is mediated by vagus.

Abstract:objective:to explore the therapeutic mechanism of tongxinluo capsule treating atherosclerosis in coronary heart disease by measuring the changes of coronary plaques and myocardial histopathology of apolipoprotein e gene knockout mice[apoe (-/-)]intervented by tongxinluo capsule. Methods:forty apolipoprotein e gene knockout mice[apoe (-/-)]were taken as the experimental group,to establish the models of coronary atherosclerosis,then randomly divided to 5 groups(model group ,simvastatin group and tongxinluo group),the laters were intervented for 8 weeks by simvastatin and tongxinluo capsule with high dose,middle dose and low dose respectively,while other 8 isogeneic mice as the normal control group. The effect and potential mechanism of tongxinluo capsule on atherosclerosis in[apoe (-/-)]mice were researched by observating the changes of some indexes in each group,such as coronary plaques and myocardial histopathology,and so on. Results:the lesions of coronary in mice of model group were mostly located at small brancha in cardiac muscle,cardiac muscle cell,myocardial interstitium,and the average score is 6.63±1.48. compared with model group,simvastatin group and different dosage of tongxinluo capsule groups had less coronary and cardiac muscle lessions,the average score of the simvastatin group was 2.62±2.25(p < 0.01),the low dose of tongxinluo capsule was 4.5±1.71(p < 0.05),the middle dose group was 3.12±1.81(p < 0.01),and the high dose group was 2.38±2.34(p < 0.01). There were no statistically significant differences between simvastatin group and high－and middle－dose tongxinluo group(p > 0.05). Conclusion:tongxinluo capsule had significant inhibition effect in coronary atherosclerosis formation in the simvastatin group compared with high－and middle－dose tongxinluo group in adose－dependent model. While the high－dose and middle－dose tongxinluo had the similar effect with simvastatin group.

Abstract:objective:to evaluate the effects of tlr3 single nucleotide polymorphisms (snps) on asthma risk and asthma－related phenotypes in a southeastern chinese han population. Methods:rs3775291,a nonsynonymous snp which was located in the fourth exon of tlr3,was selected. Three hundred and eighteen unrelated adult asthmatic patients and 352 healthy volunteers from the same area of southeast china were recruited in this study. Genotyping of the selected snp was performed using multiplex pcr in conjunction with tagged array single base extension technology. We conducted case－control association studies between the selected snp in tlr3 and asthma or asthma－related phenotypes. Results:rs3775291 snps was not associated with asthma morbid risk in whole. However,ga+aa genotypes had a significant reduced risk of atopic asthma by 31% (adjusted or= 0.69,95%ci = 0.49~0.97),which showed an independent role. The carriers of ga/aa genotype appeared less nocturnal asthma symptom compared with those of gg genotype. No correlation was found between the snp and eosinophil counts,total serum ige levels or asthma severity. Conclusion:tlr3 polymorphisms may play a considerable role in the pathogenesis of atopic asthma. It could be clinically useful both for identifying patients at risk of asthma and for preventing its occurrence.

Abstract:objective:to investigate the association between c－reaction protein (crp) gene c+1444t polymorphism and unstable angina pectoris (uap) in the chinese han population of suwan region. Methods:one hundred and forty－two patients with uap and 307 control subjects free from coronary artery disease were recruited into the study. The c+1444t polymorphism in crp gene was determined by polymerase chain reaction and restriction fragment length polymorphism analysis(pcr－rflp). Results:there was no statistical difference of the frequencies of cc,ct and tt genotype in the uap group and control group (p was 0.117,0.061 and 0.131,respectively) . The prevalence of c allele was 90.5% and 92.86%(p = 0.221) in the uap group and control group,respectively. Conclusion:the crp gene c+1444t polymorphism in the promoter region was not strongly associated with uap in the chinese han population of suwan region.

Abstract:objective:to study the qualitative and quantitative methods of single herb in anti－virus oral liquid,and improve the quality standard of anti－virus oral liquid. Methods:on qualitative study,fructus forsythiae,rhizoma anemarrhenaeand and rhizoma acori tatarinowii were identified by thin－layer chromatography(tlc),and herba pogostemonis,radix rehmanniae were also studied. On quantitative study,the method of high performance liquid chromatography (hplc) for determination of phillyrin in anti－virus oral liquid was established. Results:rhizoma anemarrhenae,rhizoma acori tatarinowii and fructus forsythiae can be identified by tlc. The average recovery of phillyrin was 99.42%,rsd =1.75%. Conclusion:the method is simple,accurate and specific,and can be used for the quality control of anti－virus oral liquid.

Abstract:objective:to study xiaoaijiedufang,which under the guidance of “cancer toxin”theory of professor zhou zhongying for long－term clinical practice as an effective anti－cancer drugs,the immune function of the chinese medicine on patients with advanced malignant tumor. Methods:one hundred and ninety－eight cases of stage Ⅲ~Ⅳ patients with malignant tumor were randomly divided into the treatment group (chemotherapy combined with chinese medicine group) of 120 cases and the control group (chemotherapy alone) of 78 cases. The patients in the treatment group took xiaoaijiedufang of chinese medicine at the begining of the first cycle of chemotherapy,and took 2 to 3 months consecutively. Results:the number of cd3+,cd4+,cd4+/cd8+ and nk activity had a significant difference between the control group and treatment group (p <0.01). Conclusion:the results indicated that xiaoaijiedufang combined with chemotherapy has a certain therapeutic effect on improving anti－tumor immunity of patients.

Abstract:objective:to study the correlation of mammographic signs to the expressions of er,pr(estrogen/progesterone recepter)and c－erbb－2 in infiltrating ductal carcinoma of breast(idc). Methods:pre－operative mammography was performed in all the 68 cases of patients with pathologically－proved idc of breast. The specimens of mastectomy of the idc were stained with immunohistochemistry,and the expression of er,pr,and c－erbb－2 were measured. Results:on the mammograms of the 68 cases,masses were shown in 54 cases(79.4%),spicule sign in 31 cases(45.6%),calcifications in 27 cases(39.4%)and breast architecture distortion only in 6 cases(8.8%). The positive expression of er,pr and c－erbb－2 were found in 41cases(60.3%),45 cases(66.2%)and 39 cases(57.4%)respectively. Er and pr positive expression was negatively correlated with the calcification of infiltrating duct carcinoma of breast(p﹤0.05),but c－erbb－2 positive expression was positively correlated with the calcification of infiltrating duct carcinoma of breast(p﹤0.05). Conclusion:the molybdenum target mammographic features of idc were closely correlated to the expressions of er,pr and c－erbb－2,and these features were in a considerable extent reflecting the biological behaviors of tumor cells and prognosis of cases. Therefore,molybdenum target mammographic features were helpful in treatment planning of breast cancer.

Abstract:objective:to know the anemia status of patients with cleft lip and palate,further analyze certain specific related factors with their anemia prevalence,as well as furnish evidence for prevention to anemia. Methods:346 patients with cleft lip and palate who had received operation in stomatology hospital affiliated to njmu from january 2007 to november 2008,were analyzed statistically. The changes of rbc and hgb between before restoration and after restoration of 12 patients whose operations were all conducted in this hospital were also investigated. Results:according to the deformity type and restoration level,346 patients with cleft lip and palate are divided into 4 groups:cleft lip and palate group (a),cleft lip group (b),cleft palate group (c) and after restoration group (d). The anemia prevalence rates of these groups are 43.6%,40.5%,19.7% and 17.0%,respectively. The anemia prevalence rates of group a and group b are all higher than that of group d,and the anemia prevalence rates of group c and group d have no difference statistically. After standardization of urban－rural and gender composition,the anemia prevalence rates of group a,group b and group c,are all higher than that of chinese population in a same age group. The anemia prevalence rate of patients whose cleft lip and palate have not been restored completely did not obviously depend on the urban－rural and gender differences. In all the patients with cleft lip and palate,there were 81 anemia patients. In the 81 anemia patients,there were 44 microcytic hypochromic anemia patients. In 12 patients,the values of hgb increased significantly and the average values of rbc were in normal range when they were restored completely. Conclusion:the cleft lip and palate enhance the possibility of anemia,and the main type of anemia is microcytic hypochromic anemia. Scientific feed fashion should be widely published in the family of the patients with cleft lip and palate,especially when the patients are young,and let them pay attention to iron－content food intake.

Abstract:objective:to explore the clinical value of mscta in the visualization of gastric arteries. Methods:dual－phase cta using a siemens sensation 16 scanner was performed in 80 cases without abdominal surgery and stomach disease. Cta of gastric arteries were made using vr,mip and tsmip techniques. The frequency and degree of visualization of the feeding arteries of stomach were recorded and evaluated by two radiologists together. Results:the left gastric artery and right gastroepiploic artery were all seen in each technique. In vr,mip,tsmip,visualization rates of right gastric artery were 50.00%,70.00%,90.00%,respectively;left gastroepiploic artery,30.00%,60.00%,77.50%;posterior gastric artery,6.25%,16.25%,25.00%;short gastric artery 10.00%,25.00%,40.00%. There were significant difference in visualization rates of right gastric artery,left gastroepiploic artery,posterior gastric artery,short gastric artery between vr and tsmip,as well as between vr and mip. There were significant difference in visualization rates of right gastric artery,short gastric artery between mip and tsmip. There were significant difference in the degree of visualization of the feeding arteries of stomach among tsmip,mip and vr. Conclusion:the gastric arteries are displayed better on tsmip than on vr and mip. The gastric arteries have three－dimensional sense of space on vr .the anatomic features and the variations of gastric arteries can be accurately displayed by vr and tsmip.

Abstract:objective:to study the effects of different anesthetic techniques on neonate perioperative stress response in the radical operation of hirschsprung’s disease. Methods:forty asaⅠ~Ⅱ neonate patients were randomly assigned into ga group and cb group with 20 each. Anesthesia in ga group was induced with fentanyl,midazolam,rocuronium and maintained with sevoflurane inhalation. Caudual block was performed in cb group after induction. Blood samples were taken before induction of anesthesia (t1),at beginning of operation (t2),30 min after incision(t3),30 min after operation(t4) and 24 hours after operation(t5). Serum levels of cortisol and interleukin－6(il－6) were measured. Systolic blood pressure(sbp),diastolic blood pressure(dbp) and heart rate(hr) were recorded at the same time. Results:in ga group,hr,sbp and dbp were significantly higher at t4 than those at t1 (p < 0.05). Hr,sbp and dbp were significantly higher at t4 in ga group than those in cb group(p < 0.05).compared with those at t1,the concentrations of cortisol at t4,5 were significantly higher in ga group(p < 0.05). The concentrations of cortisol at t4,5 in ga group were significantly higher than those in cb group(p < 0.05). Compared with those at t1,the concentrations of il－6 at t3-5 were significantly higher in both groups(p < 0.05). The concentrations of il－6 at t4,5 in ga group were significantly higher than those in cb group(p < 0.05). Conclusion:severe perioperative stress response may occur in neonate. General anesthesia combined with caudual block is superior to general anesthesia in alleviating the neonate perioperative stress response.

Abstract:objective:to investigate the therapeutic effect and post－operative complications of modified ileal orthotopic neobladder after radical cystectomy. Methods:radical cystectomy and ileal neobladder were applied in 21 male patients with improved surgical method. Results:twenty－one cases of ileal neobladders have been followed up for 1 to 4 years with a mean of 2.1 years. The operation was taken 5.5 to 7.5 hours with a mean of 5.9 hours. The intraoperative blood transfusion was 150 ml to 1 200 ml with a mean of 450 ml. All patients achieved excellent follow－up. The mean bladder capacity of the entire group was 330 ml,and residual urine volume was less than 80 ml. No patient suffered for electrolytical disturbance and urethral tumour recurrence. Conclusion:the modified radical cystectomy and ileal neobledder was a perfect surgery artifice.