Objective To compare the clinical effect of penehyclidine hydrochloride and atropine in the treatment of acute organophosphorus pesticide poisoning.Methods 86 patients with organophosphorus pesticide poisoning were selected and randomly assigned to the observation group and the control group according to the digital table,43 cases in each group.The two groups were given conventional treatment,while the control group was treated with atropine,the observation group was given penehyclidine hydrochloride.The disappearance time of main symptoms, rescue success rate and adverse reactions of the two groups were observed.Results After salvage treatment, the rescue success rate of the observation group was 97.8%,which of the control group was 88.4%,the difference was statistically significant (x2=1.433,P<0.05).The disappear time of M like symptoms,N like symptoms and central nervous system symptoms in the observation group was significantly shorter compared with the control group (all P<0.05).The number of drugs,dosage,cholinesterase recovery time and hospitalization time between the two groups had statistically significant differences (P<0.05).The incidence rates of blurred vision,restlessness,heart rate and urinary retention in the control group were significantly higher than those in the observation group (all P<0.05).Conclusion Penehyclidine hydrochloride for acute organophosphorus pesticide poisoning can significantly reduce the incidence of symptoms,shorten the disappearance time of symptoms,reduce hospitalization time,improve the efficacy of rescue,reduce the incidence of adverse reactions, it is safe,effective and has great clinical significance.

Objective To investigate the role of Wnt/β-Catenin signaling pathway in the endotoxin induced acute lung injury (ALI) during the treatment by mesenchymal stem cells (MSCs).Methods Six SPF male SD rats were isolated and bone marrow mesenchymal stem cells were cultured.A total of 72 SPF male SD rats with 6-week-old were randomly (random number) divided into 4 groups:control group (n =18) in which phosphate buffered solution (PBS) used instead of lipopolysaccharide (LPS);LPS group (n =18) in which LPS used to induce acute lung injury;LPS + MSCs group (n =18) in which MSCs directly transplanted after injection of LPS;Control + MSCs group (n =18) in which MSCs transplanted after injection of PBS.And then 6 rats of each group were sacrificed at 6 h,24 h,and 48 h separately after injection of LPS.At 24 h after the modeling,lung tissue was taken and the levels of Wrnt signaling pathway components were detected by using immunohistochemistry and Western blot.In addition,quantitative realtime PCR was used to detect the expression of Wnt signaling pathway target genes.Results Compare with the PBS control group,significant decrease in lung dry-to-wet ratio and increase in arterial oxygen partial pressure (PaO2) were found in MSCS transplantation groups.According the immunohistological results,Wnt 5a was significantly increased in the LPS-induced ALI rats and decreased after MSCs transplantation.Moreover,decrease in levels of GSK-3β phosphorylation and β-catenin was found in the lung tissue after MSCs transplantation.In addition,the expressions of Wnt signaling target genes Vegf,Axin2 and Klf4 were decreased significantly after MSCs transplantation.Conclusions In the setting of ALI,the therapeutic effect of MSCs was exerted by decreasing the expressions of Wnt 5a,GSK-3β phosphorylation,β-catenin,and Wnt signaling target genes Vegf,Axin2 and Klf4.Wnt signaling implicated in the therapeutic effect of MSC in the setting of ALI.

Objective To investigate the effects of trichostatin A (TSA) on cell proliferation and cell cycle in human gastric cancer cell line SGC-7901 in vitro and its mechanism. Methods SGC-7901 cells were treated with 0.1, 0.5 and 2.0 μmol/L of TSA for 24 hrs. Growth inhibition rates of cells were measured by MTT assay and cell cycles were detected by flow cytometery (FCM). Expressions of cyclin D1 and p21 mRNA were measured by real-time PCR. Results The proliferations of SGC-7901 cells were inhibited when treated with TSA for 24 hrs. The inhibition rates in groups treat with 0.1, 0.5 or 2.0 μmol/L of TSA were 3.52%±6.11%, 13.29%±4.13% or 14.24%±2.80% ,respectively. The cell percentage of G0/G1 phase were higher in 0. 5 pznol/L group (71.26%±0.51%) and 2.0 μmol/L group (71.03%±0.12%) compared with control group (51.12%±1.17%). The cell percentage of S phase were lower in 0.5 μmol/L group (13.55%±0.44%) and 2.0 μmol/L group (10.63%±0.63%) compared with control group (34.60%±0.60%). The expression of cyclin D1 mRNA was down-regulated, whereas p21 mRNA expression was up-regulated. Conclusions TSA inhibits SCG-7901 gastric cancer cell proliferation by affecting the cell cycle control gene eyclin D1 and p21 mRNA expressions, which induce G0/G1 cell phase cycle arrest and ultimately impact on the growth of tumor cells.

Objective To investigate the early risk factors of the formation of pancreatic pseudocysts after severe acute pancreatitis. Methods One hundred patients with severe acute pancreatitis admitted from Jul. 2005 to Mar. 2007 were included. Clinical and laboratory data within 24 hours of admission and radiological tests of chest, abdominal dynamic contrast-enhanced computed tomography and abdominal ultrasound within 3 days after admission were analyzed and multiple stepwise logistic regression analysis was performed. Results 30 patients developed pancreatic pseudocysts and the incidence of pancreatic pseudocysts in the clinical course of severe acute pancreatitis was 30%. There were significant difference between group A (pancreatic pseudocyst group) and group B (non-pancreatic pseudocyst group) in serum albumin[(33.23±4.810g/L vs (36.07±4.92)g/L], CT severity index (CTSI) (3~6 vs 2~4 points) ,length of hospital stay[(26.83±19.760) day vs (14.51±7.71) days, (P<0.05)]. Meanwhile, there were no significant differences between the two groups in age, gender proportion, body temperature, heart rate, breath rate and mean arterial pressure in admission,urine volume within 24 hours, early defaecation within 24 hours after admission, blood routine, liver function, kidney function, electrolytes, blood cholesterol and triglycerol, PT, APTr, arterial blood gas analysis, blood amylase, C-reaction protein, APACHE Ⅱ, RANSON scoring, early ascites and pleural effusion. But multiple stepwise logistic regression analysis showed that the serum albumin and CTSI were associated with the formation of pancreatic pseudocysts after severe acute pancreatids. Conclusions The serum albumin and CTSI were the independent risk factors of the formation of pancreatic pseudocysts after severe acute pancreatitis.

Objective To investigate effects of histone deacetylase 1 (HDAC1) gene silencing on proliferation and apoptosis of pancreatic cancer cell line PaTu8988. Methods The PaTu8988 cells were cultured and divided into control group (untreated), negative control group (treated with 30 nmol/L negative siRNA), low HDAC1 group (treated with 15 nmol/L HDAC1 siRNA) and high HDAC1 group (treated with 30 nmol/L HDAC1 siRNA). The real-time PCR and Western blot were used to detect the efficiency of HDAC1 gene silencing on mRNA and protein levels ,respectively, at 48 hours after transfection of HDAC1 siRNA. Cell proliferation and apoptosis were evaluated using cell counting kit and flow cytometry, respectively. Results Forty-eight hours after transfection of HDAC1 siRNA, the expression of HDAC1 mRNA level in PaTu8988 cells was 46.1%±6.1% in low HDAC1 group and 32.3%±1.4% in high HDACI group, which were lower than that in control group (100.0%±3.4%) and negative control group (87.4%±28.3%,P<0.05). The expression of HDAC1 protein was higher in control and negative control groups than in low and high HDAC1 groups. Cell survival rate was 100.0%±17.1% in control group, 87.1%±5.0% in negative control group, 68.7%±4.7% in low HDAC1 group and 61.6%±2.0% in high HDAC1 group with significant difference (P<0.05). Meanwhile, the cell apoptic rate in control (4.20%±0.95%) and negative control (4.59%±1.26%) groups was lower than that in low (10.09%±1.36%) and high (11.19%±6.07%) HDACI groups (P<0.05). Conclusions HDACI siRNA can effectively and specifically inhibit the expression of HDAC1 and proliferation of PaTu8988 cells and induce cell apoptosis.

Objective To investigate the effects of histone deacetylase 1 (HDAC1) gene silencing on cell cycle of pancreatic cancer cell line PaTu8988 and possible mechanism.Methods The PaTu8988 cells were routinely cultured and divided into control group,negative control siRNA group (c-siRNA),HDAC1 siRNA 15 nmol/L group and HDAC1 siRNA 30nmol/L group.After Liposomes 2000 transfection for 48 h,the Western blotting was used to detect the efficiency of HDAC1 gene silencing on protein levels and the expression of p21 protein.Cell cycle was evaluated by using flow cytometry.Results Compared with that of control group,the expression of HDAC1 protein in PaTu8988 cell of HDAC1 siRNA 30nmol/L group was significantly decreased,and the expression of p21 protein was significantly increased; the percentage of G2/M phase cells were significantly decreased[(21.48 ±3.67)％ vs.(28.28 ±2.94) ％,P＜0.05]; while the percentage of S phase cells were significantly increased[( 50.20 ± 6.85 ) ％ vs.( 32.49 ± 2.78 ) ％,P ＜ 0.05].Conclusions HDAC1 siRNA can efficiently and specifically inhibit the expression of HDAC1 in PaTu8988 cells,and can induce S phase cells arrest,which may be related with up-regulation of p21 protein expression.

Objective To investigate the clinic significance of HDAC1 expression in human pancreatic carcinoma.Methods 30 samples of pancreatic carcer tissues and paracancerous tissues were collected.HDAC1 expression was detected by real-time PCR and immunohistochemistry techniques.The relationships between clinicopathological findings and the expression of HDAC1 were analyzed. Results The RQ level of HDAC1 mRNA expression in human pancreatic carcinoma tissues and paracancerous tissues was 2.60(0.42～12.81)and 1.02(0.19～3.58),respectively(P=0.001).The percentage of positive cells expressed HDAC1 protein in human pancreatic carcinoma tissues and paracancerous tissues were(56 ±26)%and(6±6)%,respectively(P=0.000).The highly expressed HDAC1 correlated significantly with an advanced TNM stage and lymph node metastasis.Conclusions In pancreatic carcinoma tissues,highly expressed HDAC1may indicate the status of advanced TNM stage and poor prognosis.

Objective DNA methytransferase 1(DNMT1)is highly expressed in many cancers and lowly expressed in normal adult cells.This study was to assess effects of DNMT1 gene silencing on proliferation and apoptosis of pancreatic cancer cell line PaTu8988.Methods It is divided into four groups:Control group,Lipofectamine 2000 group,Negative control siRNA(N-siRNA)group(30 nM)and siRNA group(30nM).The expression levels of DNMT1 mRNA were detected by real-time PCR to assess the efficiency of DNMT1 gene silencing.Cell proliferation was analyzed by WST-8 assay;Cell apoptosis was evaluated by flow cytometry.Results Relative to Control group,48h after transfection of DNMT1 siRNA,The inhibitory rates of DNMT1 mRNA levels in PaTu8988 cells was(86.0±4.3)%.Cell survival rate was declined to 47.6±5.6%from Control group(100.7±3.0%),Lipofectamine 2000 group(64.5±6.8%),N-siRNA group(68.1±4.1%)(P＜0.05);Cell apoptosis rate was increased from Control group(7.51±1.12)%to siRNA group (14.94±2.89)%(P＜0.05),respectively,48h after transfection of DNMT1 siRNA.Conclusions DNMT1siRNA can efficiently and specifically knockdown the expression of DNMT1 mRNA and inhibit the proliferation of PaTu8988 cells,and induce cell apoptosis.It provides evidence for gene therapy of pancreatic cancer.

ObjectiveTo assess the effects of DNA methyhransferase 1 ( DNMT1 ) gene silencing on DNMTs activity and methylated CpG sites of hMLH-1 in pancreatic cancer cell line PaTu8988.Methods DNMT1 siRNA and negative control siRNA was constructed by Ambion Company of United States.Then they were transfected into pancreatic cancer cell line PaTu8988 at the concentrations of 15,30 nmol/L,and the cells without transfection was used as the control group.Real-time PCR and Western blotting were applied to detect the DNMT1 mRNA and protein expression,and DNMTs activity was detected by using DNMTs activity assay kit.Change of methylation of CpG island of hMLH-1 was detected by bisulfite sequencing PCR (BSP).The expression of hMLH-1 mRNA was detected by Real-time PCR.ResultsAt 48 h after transfection,Realtime RT-PCR analysis showed that the levels of DNMT1 mRNA in DNMT1 siRNA group ( 15 nmol/L) and DNMT 1 siRNA group (30 nmol/L) were 0.573 ± 0.026 and 0.143 ± 0.044,which were significantly lower than those in control group 1.020 ±0.217 and negative siRNA 15 nmol/L group 0.900 ±0.475,and negative siRNA 30 nmol/L group 0.938 ± 0.327 (P ＜0.05 ).Western blotting analysis showed that the level of DNMT1 protein of DNMT1 siRNA group was also lower than those of negative siRNA and control groups.DNMT activity in DNMT1 siRNA15,30 nmol/L groups was 0.364 ± 0.124and 0.250 ± 0.072,which were significantly lower than those in control group 0.931 ± 0.065and negative siRNA group 0.665 ± 0.055 and 0.472 ± 0.040.DNMT activity was positively correlated with DNMT1 mRNA expression ( r =0.69,P ＜ 0.01 ).DNMT1 RNA interference decreased 8 methylated CpG sites of hMLH-1 to 1 site.Concluslons DNMT1siRNA can specifically inhibit the expression of DNMT1 gene of PaTu8988 and DNMT activity,and can decrease methylated CpG sites of hMLH-1 gene.

Objective To investigate the effect of trichostatin A (TSA) on inhibition of cell proliferation and cell cycle of human pancreatic cancer cell line PaTu-8988 in vitro. Methods PaTu-8988 cells were treated with different concentration TSA (0.1,0.5,2.0μmol/L), and blank control group was used. The growth inhibition rates of cells were measured by WST-8 method. Cell cycle and apoptosis were detected by flow cytometry; the expressions of p21 mRNA and HDAC1 mRNA were measured by Real-Time PCR. Results Survival rates of TSA 0.1μmol/L,0.5μmol/Land2.0μmol/L group was (88.5±4.2)%, (79.7±5.O)% and (64.3±7.2)%, respectively, which were all significantly lower than (100.0±4.2)% of the blank control group (P<0.01). Cells in the group of TSA O.1μmol/L and 0.5μmol/L were hindered at G1 phase, while (50.29±7.53)% of the cells in the group of TSA 2.0μmol/L were hindered at G2/M phase. The expression rates of p21 mRNA in different TSA group were 5.29±1.16, 7.79±0.41, 8.61± 0.73, respectively, which were higher than that in the control group (1.00±0.08, P<0.01); the expression rates of Cyclin DI mRNA were 1.13±0.12, 0.42±0.06, 0.19±0.06, respectively, which were higher than that in the control group (1.00±0.07, P<0.05). Conclusions TSA may up-regulate the expression of p21 and down-regulate the expression of cyclin D1, and induce cell cycle blockade.