Objective To investigate the DNA methylation feature and DNA methylation regulation to its transcription and expression of O6-methylguanine-DNA methyltransferase gene (MGMT) in NaAsO2-treated HaCaT cells. Methods HaCaT cells were treated 72 hours at intervals and repeatedly by 3.13, 6.25,12.50, and 25.00 μmol/L NaAsO2, MGMT gene promoter region was amplified in the transcription initiation site - 329 - + 93 region by bisulfate-sequencing polymerase chain reaction (BSP), the mRNA transcription and the protein expression of MGMT was detected by real-time quantitative PCR and Western blotting. NaAsO2-untreated HaCaT cell was set as a blank control, and human epidermal squamous carcinoma cell strain A431 was set as a positive control. Results Among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, the positive rates of the DNA methylation of promoter region in MGMT gene were 0.63%(l/160), 6.25% (10/160), 10.63%( 17/160) and 18.75% (30/160), respectively, and methylated CpG sites were mainly located in - 249--146 region relative to transcription start site. There was no DNA methylation in the blank control. There were significant differences between the blank control and the NaAsO2-treated cells (x2 = 76.687, P< 0.05). Average levels of MGMT mRNA were 1.518 31 ± 0.180 54, 1.425 22 ± 0.180 39, 1.014 54 ± 0.096 79 and 0.887 72 ± 0.020 00, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells(1.198 29 ± 0.159 97), there were significant differences(F = 37.359, P < 0.05). Average levels of MGMT protein were 1.174 47 ± 0.064 75, 0.848 83 ± 0.057 01, 0.471 63 ± 0.023 34 and 0.240 34 ± 0.014 43, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells (1.066 19 ± 0.061 24), there were significant differences(F = 20.687, P < 0.05). Conclusions Arsenic can cause CpC island hypermethylation in the promoter region of MGMT gene, which results in inhibited MGMT mRNA transcription and protein expression. It might be one of the important mechanisms of arsenic-induced skin lesion.

The aim of this study was to evaluate the clinical application of fecal elastase test in exocrine insufficiency of chronic pancreatitic patieats. The fecal elastase 1 was detected by ELISA method in 55 cases with chronic pancreatitis(CP) and 25 cases with nonpancreatic digestive disease, and the urine BT PABA was measured by DACA method simultaniously.The results showed that the fecal elastase 1 and urine BT PABA excretion in patients with CP were much lower than those in patients with nonpancreatic disease ( P

The purpose of this study was to evaluate the killing effects of CD/5-FC suicide gene system mediated by adenovirus vector on human pancreatic carcinoma. Recombinant adenoviruses containing cytosine deaminase(CD) gene were constructed by the homologous recombination method in bacteria. The newly recombinated Ad-CD containing green fluorescent protein (GFP) was propagated in 293 cells and purified by cesium chloride gradient centrifugatioa Human pancreatic carcinoma cell line was infected with this virus, then 5-FC was added, and XTT assay was used to estimate relative numbers of viable cells. A heterotopic human pancreatic adenocarcinoma was successfully established in the flank of Balb/c nude mice. Adenoviral liquid containing CD gene was directly injected into xenografts following administration of 5-FC to observe the antitumor effects. Positive clones were selected by using endonuclease to digest the recombinants and the concentration of viral liquid containing CD gene was 2 1011 pfu/ml. It was found that significant cytotoxic activities were possessed by 5-FC for CD gene-transduced pancreatic carcinoma cells, but there was little effect on nontransduced ones. The anticancer effect was seen in vivo in xenografts of nude mice with in situ CD gene transduction. Our conclusian is that CD gene mediated by adenovirus has a high infectivity and is efficient for gene therapy of pancreatic carcinoma. These data demonstrate the therapeutic efficacy of an enzyme prodrug strategy in treatment of experimental pancreatic

Objective To study on the efficacy of Octreotide prophylaxis of post ERCP pancreatitis (PEP) and hyperamylasemia. Methods The study was conducted in 12 digestive endoscopic units in China. Patients were randomized into two groups. Octreotide group: Octreotide (0. 3mg) were dissolved in 500 ml of 0.9% saline solution and administrated by continues intravenous infusion, beginning 1 hr before the endoscopic examination and continuing for 6 hr afterward, 0. 1 mg Octreotide were injected subcutaneously at 6h, 12h after the intravenous injection stopped. Control group was given a placebo (saline solution) intravenously without subcutaneous injection. Results A total of 961 patients were accepted in the study, 832 patients were enrolled in the final analysis, Octreotide group 414 cases, and control group 418 cases. The overall incidence of acute pancreatitis was 3.85% (32/832) .which includec 2.42% (10/414) in Octreotide group and 5. 26% (22/418) in control group (P =0. 046). Incidence of hyperamylasemia was 14. 9% (124/ 832) which included 12. 32% (51/414) in Octreotide group and 17. 46% (73/418) in controlled group (P = 0. 041). The two groups were matched in many basic aspects, such as sex, age, contrast agent, indication of ERCP, the times of visualization of pancreatic duct and bile duct, etc. There was no side effect associated with Octreotide found. Conclusion The results of this trial indicate that Octreotide can prevent post ERCP pancreatitis and hyperamylasemia.

Objective To evaluate the killing effects of cytosine deaminase (CD) gene mediated by adeno virus vector on human pancreatic cancer cell lines in vitro. Methods After CD gene was cloned into pAdTrack CMV CD, pAdTrack CMV CD and pAdEasy 1 were recombinated in bacteria. The newly recombinated Ad CD containing green fluorescent protein (GFP) was propagated in 293 cells and purified by cesium chloride gradient centrifugation. Human pancreatic cancer cell lines Patu 8988 and SW 1990 were infected with this virus, then 5 FC was added, XTT assay was used to estimate relative numbers of viable cell. Results The positive clones were selected by using endonuclease to digest the combinatants and the concentration of viral liquids containing CD gene was 2?10 11 PFU/ml. It was found that significant cytotoxic activities were possessed by 5 FC for CD gene transduced pancreatic cell lines, but little effects on the nontransduced pancreatic carcinoma cells. Conclusions CD gene mediated by adenovirus has a high infectivity and is efficient for gene therapy of pancreatic carcinoma cell lines. These data demonstrate the therapeutic efficacy of an enzyme as a prodrug strategy in experimental pancreatic cancer.

Objective To evaluate the clinical application of fecal elastase test in exocrine insufficiency of pancreatic disease. Methods The fecal elastase 1 was detected by ELISA method in 55 patients with chronic pancreatitis, 21 with pancreatic cancer and 25 with nonpancreatic digestive disease, and the urine BT PABA was measured by DACA method simultaneously. Results The fecal elastase 1 and urine BT PABA excretion in patients with chronic pancreatitis and pancreatic cancer were much lower than those in patients with nonpancreatic disease ( P

AIM: To investigate the anti-proliferative and apoptosis-inducing effect of honokiol on U937 cell line in vitro. METHODS: Proliferation of U937 cells and PBMCs were analyzed by MTT assay. Flow Cytometry and cell morphological observation were performed to find out whether honokiol could affect cell cycle and induce apoptosis of U937 as well as PBMCs in vitro. RT-PCR and Western blotting techniques were used to detect the changes in mRNA expression and protein production of bcl-2 and bax in U937 cells after treated with honokiol. RESULTS: Honokiol could significantly inhibit the proliferation of U937 cells at IC_ 50 concentration of 11.8 ?g/mL, but slightly inhibit the proliferation of PBMCs, at IC_ 50 concentration of 40.3 ?g/mL, respectively. Most honokiol-treated cells were arrested at G_0/G_1 phase. CONCLUSION: Honokiol could inhibit the proliferation and induce apoptosis of U937 cells, while has little effect on the proliferation and survival of PBMCs. Bax might be involved in the gene regulation related to honokiol-induced apoptosis.

Objective To evaluate the clinical efficacy of multiple target burst frequency thermocoagulation in treatment of lumbar disc herniation.Methods One hundred and twelve patients suffering from lumbar disc herniation were treated with stepped heating multiple target burst frequency thermocoagulation using of domestic R-2000B radiofrequency ablation machine.The visual analogue scale (VAS) score,Oswestry disability index (ODI) and effect of grade before operation and after operation were compared.Results The VAS score before operation was (7.60 ± 1.12) scores,3 d after operation was (3.10 ± 1.05) scores,6 months after operation was (2.90 ± 0.92) scores,there was significant difference between before operation and 3 d,6 months after operation (P ＜ 0.05).The ODI before operation was 47.6 ± 8.3,3 months after operation was 25.5 ± 6.7,6 months after operation was 23.7 ± 6.2,there was significant difference between before operation and 3,6 months after operation (P ＜0.05).The clinical efficacy:excellent grade was in 66 cases,the good was in 32 cases,the improvement was in 10 cases,the inefficacy was in 4 cases,the excellent and good rate was 87.5％ (98/112).There was no nerve injury,infection or death after operation.Conclusion Multiple target burst frequency thermocoagulation in the treatment of lumbar disc herniation has the advantages of little incision and tissue damage,fast recovery,good clinical outcome,it is worth to clinical practice.

Objective To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma.Methods The expression of claudin-4 in 62 endonetrioid endometrial carcinoma (EEC),30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and realtime PCR.Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells,which were divided into three groups with different intraperitoneal treatments with cisplatin,paclitaxel and saline solution.After the observation period,the tumors were extracted and stained with monoclonal antibody against claudin-4.The messenger RNA expression of claudin-4 was also detected using real-time PCR.Results Among the EEC samples,34％ (21/62) showed medium staining for claudin-4 and 66％ (41/62) showed intense staining.In atypical hyperplasia group,27％ (8/30) showed weak staining,53％ (16/30) showed medium staining and 20％ (6/30) showed intense staining for claudin-4.Of the normal endometrial tissue,47％ (28/60) showed weak staining and 53％ (32/60) showed no staining for claudin-4.According to real-time PCR,the relative quantity of claudin-4 was 170 ± 12 in EEC group,89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium.Compared with those in atypical hyperplasia group and normal endometrium group,the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P ＜ 0.05).In the study of Ishikawa xenografts,no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group.Nevertheless,a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group.After cisplatin treatment,the tumor volume was significantly decreased [(0.51 ±0.21) versus (0.73 ±0.12) cm3],and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27).Conclusion These results demonstrate that claudin-4 is strongly expressed in EEC,which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.

Aged ; Case-Control Studies ; Cerebral Hemorrhage ; etiology ; Cerebral Infarction ; etiology ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Middle Aged ; Plasma ; chemistry ; Polymorphism, Genetic