AIM:Vascular smooth muscle cell (SMC) proliferation and migration from the arterial wall media into the intima are believed to play a critical role in the pathogenesis of restenosis. Several studies have demonstrated that phosphothioate (PS) oligodeoxynucleotides targeted against genes involved in SMC proliferation inhibits in vitro SMC proliferation and migration. However, the therapeutic effect of antisense ODN on the individual who receives the treatment of delivery of the agent depends on the efficacy of this agent in great degree. We investigated the inhibition effect of a novel agent, insulin-antisense-c-myb-PS-ODN on SMC proliferation in vitro.　METHODS：The rat aortic artery SMCs were cultured in Dulbecco's modified Eagel's medium. The passage 8 to 13 were used as the experiment. Cell surface receptor binding assay was quantified through counting gamma particles emitted from 125　　　　I labeled insulin. SMC rapid proliferation was brought by stimulation of high concentration of fetal bovine serum (FBS). The novel agent of insulin conjugated to the antisense-c-myb-PS-ODN was obtained via incubation of both in condition of certain reagents, pH, temperature, and ion concentration. The characterization and purification of the agent was performed through HPLC. Inhibition of SMC proliferation was reflected by incorporation rate of trillium labeled thymidine deoxyribonucleotide.RESULTS：The binding efficacy of insulin to the receptor was remarkably increased in SMC cultured in supplement of 20% FBS. The inhibition effect of conjugator insulin-c-myb-antisense-PS-ODN was stronger than that of the simple c-myb-antisense-PS-ODN. The inhibition rate of conjugator and simple form on SMC proliferation were 48.34% and 29.54%, respectively. CONCLUSION：The binding efficacy and specificity of c-myb-antisense-PS-ODN to SMC may be enhanced by the insulin receptor mediation through the insulin-insulin receptor interaction. The insulin-receptor targeted method may be a potential and specific therapeutic pathway for restenosis.

Objective To explore the expression of breast cancer susceptibility gene 1 (BRCA1) in human colorectal cancer and its correlation with the efficacy of platinum-based chemotherapy.Methods Seventy-eight cases of colorectal cancer patients treated with platinum-based chemotherapy after surgery were collected.The surgical specimens of them were taken.Fourteen normal colonic mucosa specimens and 12 non-cancerous tissues of colorectal cancer specimens were obtained.The expression of BRCA1 in tissues was detected by immunohistochemistry and analyzed by x2 test.The colorectal cancer patients were followed up for survival time.Comparison of survival analysis was performed by Kaplan-Meier survival curves and Log-rank test.Results The positive rate of the BRCA1 expression in colorectal cancer tissue (52.6 ％,41/78) was lower than that of para-cancer tissue (11/12,x2 =6.518,P=0.011) and normal colonic mucosa tissue (13/14,x2 =7.949,P=0.005).The poorer the differentiation of colorectal cancer,the lower the positive rate of BRCA1 (x2=14.160,P=0.001).The median disease-free survival time of BRCA1 negative colorectal cancer patients was 51.0 months (95 ％ CI:47.7 to 54.4 months),which was longer than that of BRCA1 positive patients (45.0 months,95 ％CI:36.6 to 53.4 months,x2 =4.367,P=0.032).Conclusions Receiving oxaliplatin based chemotherapy may be a survival benefit for BRCA1 negative colorectal cancer patients.The expression of BRCA1 may be an index for chemotherapy options and prognosis evaluation for colorectal cancer patients after surgery.

Objective To investigate the value of Cytochrome P450 (CYP3A5) * 3 gene polymorphism in providing individualized administration for the use of tacrolimus (Tac) in renal transplantation recipients.Method Pyrophosphate sequencing method was used to determine the CYP3A5 * 3 genotype of renal transplant patients in the first day after surgery.Sixty recipients were divided into experiment group and control group.Both groups of patients were routinely given the initial dose of Tac-4.0 mg/day in the first day after surgery.The experiment group of patients were given different doses of Tac based on the different CYP3A5 * 3 genotypes at the third day after surgery [for AA:0.12 mg/(kg· day),and for GG:0.06 mg/(kg· day)],and the control group of patients were given different dosages of Tac according to drug concentration.Different parameters were compared between two groups of patients:percentage of patients reaching the target concentration (3-8 μg/L) at the fifth day after surgery,days required to reach the target concentration level,times needed to adjust the dosage of Tac within two weeks.Result The percentage of patients reaching the target concentration in experiment group and control group was 90％ and 46.67％,respectively (P＜ 0.05).Days required to reach the target concentration were (3.67 ± 1.32) and (7.57 ± 3.42) on average,respectively (P ＜ 0.05).Times of adjusting the Tac dose in experiment group was significantly less than those in the control group (P＜0.05).In the experiment group,the target concentration was obtained even without dosage adjustment (70％).Conclusion Individualized adjustment of Tac doses for patients according to recipients' different CYP3A5 * 3 genotypes is beneficial for reaching target concentration as soon as possible,which is superior to traditional dosage regimen.

Objective To evaluate renal hemodynamic changes in hydronephrosis children with no image on intravenous urography by color doppler flow image(CDFI). Methods The resistance indexes(RIs)in main renal arteries(MRAs),interlobar renal arteries(IRAs)and arcuate renal arteries(ARAs)were measured in 48 children with hydronephrosis with no image on intravenous urography.The thickness and area were also measured in hydronephrotic kidneys.Resistive index ration(RIR=hydronephrotic kidneys RI/contralateral RI)was calculated. And pathological changes of kidneys were graded in these cases.Spearman's correlation test was used to compare RIR and pathological grades in these kidneys. Results In healthy and hydronephrotic kidneys,the relevant parameters were as fol1.13±0.14,1.14±0.09,respectively in MRAs,IRAs,ARAs in hydronephrotic kidneys(P＞ness,area of hydronephrotic kidneys(P＜0.05),also between RIR and thickness,araa(P＜0.05),MRAs,IRAs,ARAs and pathologic grades(P＜0.05),especially in IRP(s(r=0.795,P＜0.01).Conclusions RIR of IRA could be an ideal parameter of renal hemodynamics to evaluate renal damage in children with congenital hydronephrosis with no image on intravenous urography.

Objective To construct a retrovirus-mediated expression system containing double strands DNA for RNA interference on human Notch1 gene and study its inhibitory effect on human glioma cell lines U251 and CHG-5 in vitro.Methods A recombinant retroviral vector pSiRNA-Notch1 was generated by cloning a double strands DNA for RNA interference on human Notch1 gene into a retroviral vector Psilencer 5.1-H1 Retro.Human glioma cell lines U251 and CHG-5 were infected with the viral supernatant from the PT67 clones.After 3 d,the viability,Notch1 mRNA and protein of the transfected cells were examined by WST-8 assay,RT-PCR and Western blot.Results The pSiRNA-Notch1 recombinant retroviral vector had been constructed correctly.The titer assayed on NIH3T3 cells was up to 224?104 cfu/ml.Three days after transfection,the viability,Notch1 mRNA and protein of the transfected cells decreased significantly.Conclusion The constructed pSiRNA-Notch1 retroviral vector shows effective inhibition effect on the viability in human malignant glioma cells,with potential utility in the gene therapy for human malignant glioma.

Objective To investigate the feasibility and efficacy of laparoscopic orchiopexy for inguinal palpable undescended testes. Methods Ninety patients with 103 inguinal undescended pal-pable testes were treated by laparoscopic orchiopexy performed by the same surgeon. There were 24 (26.7 %) left and 53 (58. 9%) right palpable cryptorchidism cases, plus 13 cases (14.4 % ) with bilat-eral undescended testes. The mean age of the patients was 17 months (range 8 months-6 years). The surgical procedures were described as following. First, the peritoneum was opened at the anterior sur-face of the spermatic vessels and vas deferens before the testis was pulled into the abdominal cavity so that the spermatic vessels and vas deferens were released and easily manipulated. Second, the inter-space between internal spermatic fascia (transversalic fascia) and processus vaginalis was divided so, that the testis could be pulled into the abdominal cavity. Finally, the trocar which was inserted from scrotum was placed through the outer ring. Finally, the testis was placed at position and fixed. Re-suits The mean operative time was 32.7±5.2 min. The processus vaginalis unclose was found in 93 (90.3%) cases; and contralateral process us vaginalis unclose were found in 12 (15.6%) cases of 77 unilateral undescended testes. The complication rate was 3(3.3%)cases, all in bilateral cases. All 103 testes were descended by laparoscopy. On follow-up ranging 6-12 months, all testes maintained good size and a proper position. Conclusions The laparoscopic approach may be a safe way to descend the inguinal palpable testes. Orchiopexy of palpable undescended testes can be done with advantages in the laparoscopic approach.

To observe the effects of diabetogenic (high fat high sucrose, lacking choleserol) diet on atherogenesis in New Zealand white rabbits. Two groups of New Zealand white rabbits received regular rabbit chow (the normal control), or high fat high sucrose diet for 4 months. The levels of plasma total cholesterol, HDL cholesterol, triglycerides, insulin, and glucose were investigated, the areas of fatty streak of the aortae were measured after staining with Sodan IV, and the aortic, coronary specimens were observed with light and electron microscopies. The plasma glucose, triglycerides, and total cholesterol were increased significantly by high fat high sucrose feeding. At the end of 4 months, the early charateristics of atherosclerosis were present in the animals' vascular specimens. Our findings suggest that high fat high sucrose feeding can induce hyperglycemia, hypertriglyceridemia and atherosclerosis in New Zealand white rabbits, and this could be a potential animal model for studying the mechanisms of diabetes-accelerated atherosclerosis. This study raised a question: What is the mechanism by which high fat high sucrose feeding induces atherosclerosis?. The related hypothesis was given in this article.

AIM: To study the action of ATP binding cassette transporter(ABC) A 1 on cholesterol efflux in THP-1 macrophage-derived foam cells. METHODS: After exposure of the cultured THP-1 macrophage-derived foam cells to 22(R)-hydroxycholesterol and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) at different concentration for 24 hours, cholesterol efflux and ABCA1 mRNA level were determined by FJ-2107P type liquid scintillator and reverse trancriptase-polymerase chaim reaction(RT-PCR), respectively. RESULTS: Oxidized LDL promoted cholesterol efflux in THP-1 macrophages and 22(R)-hydroxycholesterol increased cholesterol efflux in THP-1 macrophage-derived foam cells in a dose-dependent manner and DIDS inhibited cholesterol efflux in THP-1 macrophage-derived foam cells in a dose-dependent manner. Exposure of the cultured THP-1 macrophage-derived foam cells to 22(R)-hydroxycholesterol and DIDS at different concentration for 24 hours, resulted in increase and decrease in the expression of ABCA1 mRNA in THP-1 macrophage-derived foam cells in a dose-dependent manner, respectively. CONCLUSION: ABCA1 playes an important role in cholesterol efflux in THP-1 macrophage-derived foam cells.

AIM: Anti-atherosclerosis effects of Momordica charantia L was further studied in a New Zealand rabbit atherosclerotic model at the basis of anti-inflammation and antioxidant effects. METHODS: Animals were divided into 3 groups: normal group (normal rabbit diet), atherosclerosis group(diet containing 2% cholesterol), and Momordica charantia L group(diet containing 2% cholesterol and 1 5% sarcocarp of Momordica charantia L ). Ninety days later, all animals were sacrificed. The effect of Momordica charantia L on atherosclerosis was evaluated by measuring serum lipid and total cholesterol content of artery wall, observing fatty liver degree, aorta arteriosclerotic area, and the thickness of intima. RESULTS: The level of total serum cholesterol and LDL-C in Momordica charantia L treatment group were obviously lower than those in atherosclerosis group, so were the total cholesterol content of artery wall, fatty liver degree, atherosclerotic area, intima thickness and I/M ratio, but no significantly difference was found between the two groups in TG level. The level of HDL-C in Momordica charantia L treatment group was evidently lower than that in normal control group. CONCLUSION: Momordica charantia L has an anti-atherosclerosis action in rabbits.

AIM: To investigate the relationship betw een ADRP and the development of atherosclerosis. METHODS: Antisense oligodeoxynucleotide of mouse ADRP was constr ucted. The mouse peritoneum macrophages were cultured with Ox-LDL or Ox-LDL plus the antisense fragment. The cellular cholesterol was measured and the expressio n of ADRP was observed with RT-PCR and western blotting. New Zealand white rabbi ts were fed with high cholesterol chow for 12 weeks. The levels of serum lipid a nd cholesterol content of aortic wall were investigated. The areas of fatty stre ak of the aortas was measured after staining with Sudan Ⅳ. The aortic, and live r specimens with HE and immunohistochemistry staining were observed under light microscopes. RESULTS: Antisense oligodeoxynucleotides of mouse ADRP decreased cellular cholesterol ester, induced cellular lipid droplets and the expression of ADRP. The expression of ADRP was induced by high-cholesterol-diet feeding in rabbit atherosclerotic lesions. The fatty streak of the aorta with immunohistoch emistry staining was strongly positive for ADRP in animals fed with high cholest erol chow, and the liver was negative with or without high cholesterol chow. CONCLUSIONS: The expression of ADRP in vessel walls is related t o the atherosclerosis, and has a potential role in lipid accumulation in macroph ages and pathogenesis of atherosclerosis.