Purpose: To study the mechanism of chemoprevention of colorectal cancer by aspirin. Methods: Different concentrations of aspirin have been used in the culture of colonic cell-Caco-2, its proliferation and apoptosis was observed during the culture and the expression of Fas/FasL mRNA was assayed by RT-PCR. Results: Major portion of cells died in the culture medium that was added high concentration of aspirin, but the lower concentration promoted apoptosis. The Fas mRNA was expressed in cells added with 25 mmol/L aspirin, but not in that of 1640. The FasL mRNA expressed in 25mmol/L aspirin was lower than in 1640. Conclusions: The lower concentration of aspirin can promote apoptosis of coloniccell, the mechanism has correlation with induction of the expression of Fas, and down-regulation of the expression of FasL.

Background and purpose:Gallbladder carcinoma, lacks of specific clinical manifestations,and is usually diagnosed at advanced stages of the diseases and even fewer can be resected.Chemotherapy has not shown significant activity in gallbladder carcinoma,so new agents should be applied to its treatment.Arsenic trioxide,verified as a breakthrough in the management of acute promyelocytic leukemia,has been applied in the research of a variety of solid tumors.This study is to investigate the biological effect of As_(2)O_(3) on human gallbladder carcinoma cell GBC cells and its mechanism.Methods:Hoechest33258 staining,DNA gel electrophoresis and flowcytometric were used to determine apoptosis.Western blot was performed to analysis apoptotic proteins expression.Results:The treatment of As_(2)O_(3) in gallbladder carcinoma cells could induce apoptosis in a dose-dependent manner and downregulate the expression of anti-apoptotic protein Bcl-2.Moreover,Bcl-2 overexpression could protect gallbladder carcinoma cells from As_(2)O_(3)-induced apoptosis.Conclusions:As_(2)O_(3) induces gallbladder carcinoma cell apoptosis via down regulation of Bcl-2.

Purpose　The aim is to establish a simple method to extract superoxide dismutase(SOD) from pig blood red cells.Methods　A combined method-microwave heating and adding Cu2+ into the suspension-was used to extract SOD from the red cells. Results　The undesired proteins were denatured by microwave heating and SOD was partly purificated in comparison with hemolysis. The processing time of the former was much shorter than the latter. Conclusion　A novel cell disruption microwave heating, was a rapid and effective technique for the primary extraction of SOD.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of C-terminal Src kinase (Csk) in Ang Ⅱ-infused rat model and cultured podocytes,and to explore the role of Csk in Ang Ⅱ-induced cytoskeletal rearrangement of podocytes.Methods Twenty-four Wista rats were randomly subjected to normal saline infusion,or Ang Ⅱ infusion at 400 ng · kg1 · min-1 (via subcutaneous osmotic minipumps) for 2 or 4 weeks.Renal histomorphology was evaluated through electron microscopy.The expression of glomerular Csk was analyzed by immunofluorescence and Western blotting.In vitro,conditionally immortalized mouse podocytes were cultured and treated with Ang Ⅱ doses ranging from 10-9 mol/L to 10-5 mol/L and for different hours.The expression of podocytes Csk was assessed by Western blotting.After transfection to podocytes with Csk siRNA,FITC-conjugated phalloidin was used to stain F-actin,to investigate the role of Csk in Ang Ⅱ-induced or cytochalasin D-induced cytoskeletal rearrangement.Results (1) Examination of Ang Ⅱ infusion rats glomerular and podocyte ultrastructure by electron microscopy revealed foot process effacement and fusion; (2) In Ang Ⅱ infusion rats,the expression of glomerular Csk was increased (P ＜ 0.05); (3) In vitro,Ang Ⅱ-stimuli up-regulated the expression of Csk (P ＜ 0.05),and the effects of Ang Ⅱ were on dose-dependent and time-dependent manner; (4) Ang Ⅱ-induced disruption of F-actin was alleviated by Csk siRNA transfection in cultured podocytes; furthermore,cytochalasin D depolymerized the F-actin cytoskeleton,while Csk siRNA stabilized the actin filaments.Conclusion The enhanced expression of Csk may be involved in Ang II-induced podocytes cytoskeletal rearrangement and foot process fusion.

OBJECTIVE:To optimize the preparation technology of Compound bovis calculus sativus gel. METHODS:The ul-trasonic emulsifying technology was optimized by orthogonal test using ultrasonic power,ratio of ultrasonic time to interval time, total ultrasonic time as factors,using centrifugal stability constant(KE)as index.Ultrasonic emulsifying method was applied to pre-pare O/W emulsions using paeonol,berberine hydrochloride and eucalyptus oil;then calculus bovis sativus powder was added into O/W emulsions,and then mixed with carbomer(940)gel matrix to prepare gel. The formulation of gel was optimized by orthogo-nal test with the amount of carbomer (940),glycerool and triethanolamine as factors,using compactibility score,comprehensive score of release rate in vitro as index. Validation test,stability test and content determination of bilirubin were conducted for gel pre-pared by optimized technology. RESULTS:The optimal ultrasonic emulsifying technology was as follows as ultrasonic power 450 W,ratio of ultrasonic time to interval time 2:1,and total ultrasonic time 5 min. The optimal formulation of gel was as follows as carbomer(940)0.5%,glycerool 15%,triethanolamine 0.20%(g/100 g). The average of KE of validation test and average compre-hensive score were 0.175 and 98.67(RSD<2%,n=3);the appearance of the preparation had no obvious change in stability test, and average percentage of bilirubin in labeled content was 100.8%. CONCLUSIONS:The optimal formulation and preparation tech-nology of gel is feasible,and the prepared gel is stable and controllable in quality.

Purpose:To evaluate the role of COX 2 in the carcinogenesis and progression of human colorectal neoplasms. Methods:The expression level of COX 2 in 122 colorectal neoplasm tissues(including 35 colorectal adenomas, 67 colorectal carcinoma and 23 colorectal cancer with synchoronous hepatic metastasis) was assayed by immunohistochemical methods. All specimens was analyzed by Conformation quantitative assay system, their stain strength was calculated.Results:Conformation quantitative assay showed the mean stain strength is 704.5 131.8 in colorectal adenoma, 1197.2 204.3 in colorectal cancer without hepatic metastasis and 1901.2 324.8 in colorectal cancer with synchoronous hepatic metastasis, there are significant differences among them statistically. According to the Dukes stages, mean stain strength is 1145.3 187.0 in B stage,1237.0?298.7 in C stage,1901.2 in D stage. There is statistic difference between Dukes stage D and B, C. Also our study indicated there was no relationship between age, sex, tumor location, tumor differentiation or tumor size with the level of COX 2 protein expression.Conclusions:In our test,higher COX 2 expression is seen in colorectal cancer than colorectal adenomas, colorectal cancer with synchoronous hepatic metastasis than colorectal cancer without hepatic metastasis,and Dukes D stage than the B?C. Thus COX 2 responses is important in the carcinogenesis and progression of colorectal cancer. NSAIDs or the others may play a role in the chemoprevention strategies of colorectal neoplasm as the COX 2 inhibitor.

Currently endoscopic retrograde cholangiopancreatograhp(ERCP) has been one of the major treatments to diagnose and cure pancreatic and choledochal diseases in many large and medium-sized hospitals.To meet the requirements of the army,ERCP treatment is practiced in field battle X-ray diagnosis vehicle(Model XCY2002-1/200) and has achieved satisfactory effects.The functions of field battle X-ray diagnosis vehicle in model of XCY2002-1/200 are expanded and new access are explored to cure patients with pancreatic and choledochal diseases in the field.

To construct a bait expression vector containing the duck circovirus Cap gene for use in the yeast two-hybrid system, the whole cap codon-optimized gene was inserted into pGBKT7 vector and confirmed by PCR, restriction enzyme digestion, and sequence analysis. After transformation into a Y2HGold yeast strain, the expression of Cap protein was analyzed by Western blotting. Toxicity and self-activation of the bait protein were detected using different dropout minimal base. PCR reaction, restriction enzyme digestion, and sequencing analyses indicated that the duck circovirus Cap gene was correctly inserted into pG- BKT7. Western blotting showed that the whole Cap protein was expressed. The recombinant bait protein had no toxicity and self-activation. Therefore, the bait vector with the Cap gene was constructed successfully, providing a foundation for future screening for interacting proteins in the yeast two-hybrid system.
Animals ; Capsid Proteins ; genetics ; metabolism ; Circovirus ; classification ; genetics ; isolation & purification ; Cloning, Molecular ; Ducks ; Genetic Vectors ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Two-Hybrid System Techniques

Objective To investigate the effect of aloe polysaccharides (AP) pre-emptive treatment on the expression of nuclear factor kappa B (NF-κB),ntercellulor adhesion molecule-1 (ICAM-1) and cell apoptosis in hippocampal brain tissue in rats with severely hemorrhagic shock for the first time of entering high altitude.Methods Forty healthy male SD rats weighing 250-300 g were randomly (random number) divided into 5 groups (n =8 each):sham group,shock group and AP group which was further divided into 3 subgroups as per different dosages of AP administered (AP1:0.75 mg/kg; AP2:1.50 mg/kg; AP3:3.00 mg/kg).Rats in sham group were treated with surgical procedure without exsanguination.Rats in shock group were exsanguinated until hemorrhagic shock emerged without resuscitation.Rats in AP subgroups were intravenously infused with given doses of AP in different AP subgroups at 30 min before hemorrhagic shock.MAP was dropped to (35 ±5) mmHg (1 mmHg =0.133 kPa) in 15 min by bleeding from femoral artery,the mean arterial pressure (MAP) was maintained at (35 ±5) mmHg for 60 min with bleeding or re-transfusing.At 3 h after resuscitation,rats were sacrificed immediately by bleeding,and the hippocampus of brain was harvested on the ice.The expressions of NF-κB and ICAM-1 in the hippocampus of rats were determined by immuno-histochemical method,and number of cell apoptosis in the hippocampus of rats was determined by TUNEL.The means were compared with analysis of variance and Student-NewmanKeuls test,and statistical significance was established at a P value of less than 0.05.Results Compared with sham group,the expressions of NF-κB (5.03 ±0.42),ICAM-1 (4.14 ±0.29) and number of cell apoptosis (44.3 ± 7.2) in hippocampal tissue were significantly increased in shock group (P ＜ 0.05).There were no significant differences in these three variables between shock group and AP1 group.Compared with shock group,the expressions of NF-κB (3.12 ±0.34),ICAM-1 (2.93 ±0.21) and number of cell apoptosis (24.8 ± 3.6) in hippocampal tissue were significantly decreased in AP2 group (P ＜ 0.05).There were no significant differences in these three variables between AP2 and AP3 groups.Conclusion AP pre-emptive treatment can significantly attenuate the expressions of NF-κB,ICAM-1 and number of cell apoptosis in hippocampal tissue in hemorrhagic shock rats.

Objective To explore the risk and treatment of laparoscopic cholecystectomy(LC) in patients with liver cirrhosis and cholelithiasis.Methods To summarize the clinical data of 28 patients with liver cirrhosis and cholelithiasis.The patients were repeated right upper quadrant pain,including 4 cases of gallbladder neck stones incarcerated,gallbladder effusion.Among them,there were 20 cases of hepatitis B cirrhosis,5 cases of schistosomiasis cirrhosis,3 cases of alcohol.Results There were 6 patients with hemorrhage during operation.Among them,5 patients were treated with gelatin sponge,hemostatic gauze and bio-glue spray to stop bleeding,1 case was transferred to open surgery because of bleeding.The remaining 27 cases of LC were successful.Complications occurred in 8 patients,3 cases of increased liver function abnormalities,1 case of upper gastrointestinal bleeding,1 case of mild hepatic encephalopathy,2 cases of significant ascites formation,1 patient underwent subtotal resection of the gallbladder with Hartmanns bag,and bile was found in the ascites after surgery,but the bile disappeared after five days.All patients with complications after symptomatic treatment were cured,no death,no major bleeding and liver failure,bile duct injury,severe biliary fistula and other serious complications.Conclusion Although the risk of LC in patients with liver cirrhosis is higher than that in the general population,with the help of current high-tech surgical instruments,we can safely accomplish this with an improved surgical procedure.As these patients require high technical requirements of surgery,surgeons must have a wealth of experience and familiar laparoscopic liver and gallbladder anatomy.