Zou Shengquan

The relationship of CyclinD1 and estrogen receptor expression in the process of proliferation and metastasis in breast neoplasm.

SummaryThe role of CyclinD1 and estrogen receptor (ER) in the process of proliferation and metastasis of breast neoplasm and their relationship were studied. The expression levels of CyclinD1 and ER in the tissue samples were detected by using flow cytometry and L SAB immunohistochemistry staining, respectively. The results showed that CyclinD1 and ER expression levels in breast cancer were significantly higher than in benign breast neoplasm (P<0.05). The CyclinDl expression levels in stage I was much lower than in stages II, III, IV (P<0.05). The positive rate of ER was not related with tumor size, lymph node metastasis and TNM stage (P>0.05), but the CyclinD1 expression level in ER (+) group was significantly higher than in ER (-) group (P<0. 05). It was concluded that CyclinDl expression level might be obviously related with the proliferation and metastasis of breast neoplasm and ER.The role of CyclinD1 and estrogen receptor (ER) in the process of proliferation and metastasis of breast neoplasm and their relationship were studied. The expression levels of CyclinD1 and ER in the tissue samples were detected by using flow cytometry and L SAB immunohistochemistry staining, respectively. The results showed that CyclinD1 and ER expression levels in breast cancer were significantly higher than in benign breast neoplasm (P < 0.05). The CyclinD1 expression levels in stage I was much lower than in stages II, III, IV (P < 0.05). The positive rate of ER was not related with tumor size, lymph node metastasis and TNM stage (P > 0.05), but the CyclinD1 expression level in ER (+) group was significantly higher than in ER (-) group (P < 0.05). It was concluded that CyclinD1 expression level might be obviously related with the proliferation and metastasis of breast neoplasm and ER.

Celecoxib inhibits proliferation and induces apoptosis via cyclooxygenase-2 pathway in human pancreatic carcinoma cells.

SummaryIn order to evaluate the effects and mechanisms of celecoxib in inhibiting proliferation and inducing apoptosis on human pancreatic carcinoma cells, the anti-proliferative effect was measured by using methabenzthiazuron (MTT) assay. Cell cycle and apoptosis were analyzed by using flow cytometry (FCM), and the PGE2 levels in the supernatant of cultured pancreatic carcinoma cells were quantitated by enzyme-linked immunoabsordent assay (ELISA). Our results showed that celecoxib suppressed the production of PGE2 and inhibited the growth of JF-305 cells, and the anti-proliferative effect of celecoxib could be abolished by addition of PGE2. FCM revealed that celecoxib could inhibit proliferation and induce apoptosis by G1-S cell cycle arrest. It was concluded that cyclooxygenase-2 specific inhibitor celecoxib could inhibit proliferation and induced apoptosis of human pancreatic carcinoma cells via suppression of PGE2 production in vitro.

Analysis of etiology and drug resistance of biliary infections.

SummaryThe bile was collected from fro patients with biliary infections, with the bacterium isolated to study the sensitivity of each kind of the bacterium to several antibiotics in common use. Except G bacterium, we also found some kinds of G− bacterium in infection bile, G− bacterium were not sensitive to Clindamycin. G− bacterium were sensitive to Ciprofloxacin. Escherichia coli. Xanthomonas maltophilia. Enterobacter cloacae. Pseudomonas aeruginosa were sensitive to Ampicillin, G− bacterium were not sensitive to Azactam. Enterococcus faccalis. Enterococcus faecium. Enterobacter cloacae were not sensitive to Ceftazidime. Enterococcus faccalis. Staphylococcus coagulase negative. Staphylococcus epidermidis. Pseudomonas aeruginosa were not sensitive to Ceftriaxone Sodium. We didn’t found any bacterium resistance Imipenem. The possibility of the existence of G− bacterium as well as drug resistance should be considered n patients with biliary infections. The value of susceptibility test should be respected to avoid drug abuse of antibiotics.

The bcl-2 mRNA Expression in GCDC-induced Obstructive Jaundice in Rats and Its Implication in Hepatocellular Apoptosis

SummaryThe modulatory role of bcl-2 gene in hepatocellular apoptosis of rats with glycochenodeoxycholate (GCDC)-induced obstructive jaundice was investigated. The hepatocytes in normal rats and those with bile duct-ligation for 7 days, 14 days and 21 days were isolated and obtained by in situ collagenase perfusion and primary culture. The expression of bcl-2 mRNA in the hepatocytes was detected by RT-PCR. Primary culture was performed on the hepatocytes from normal rats and those with bile duct-ligation for 14 days. 100 μmol/L GCDC was added to the hepatocytes for incubation for 24 h. The hepatocellular apoptotic ratio was measured by using FCM and hepatocellular apoptosis detected in situ by using TUNEL technique. Results showed that the expression of bcl-2 mRNA was not detectable in the hepatocytes of normal rats by RT-PCR technique, while detectable in the hepatocytes of those with bile duct ligation (BDL) for 7, 14 and 21 days. Hepatocellular apoptosis in the BDL group was obviously decreased as compared with normal control group after addition of 100 μmol/L GCDC to the cells for 24 h. It was concluded that the hepatocytes in the BDL rats expressed bcl-2. During obstructive jaundice, expression of bcl-2 from the hepatocytes can inhibit the bile salt-induced hepatocellular apoptosis.

Influence of Connexin43 on the Bystander Effect Induced by Double Suicide Genes System in Vitro and in Vivo

AbstractObjective: To observe the influence of connexin 43 (Cx43) on the bystander effect induced by cytosine deaminase (CD) and herpes simplex virus thymidine kinase (HSV-tk) coexpression suicide genes system in human cholangiocarcinoma QBC939 cells and transplantation tumors in nude mice. Methods: In vitro, the CD+tk+ and CD+tk+Cx+ cells were respectively treated with 5-fluorocytosine (5-Fc) and Ganciclovir (GCV). The cytotoxic effect was evaluated by MTT method. In order to investigate the influence of Cx43 on the bystander effect, the size of transplantation tumors of the CD+tk+ and CD+tk+Cx+ cells was measured before and after application of 5-Fc and GCV. Results: CD and tk genes were stably expressed in transfected QBC939 cells. The increased expression of Cx43 was determined by testing for the presence of Cx43 mRNA by RT-PCR and the presence of Cx43 protein by Western Blot in CD+tk+Cx+ cells. The killing effect of 5-Fc and GCV on CD+tk+Cx+ cells was more effective than that on CD+tk+ cells both in vitro and in vivo. Conclusion: Double suicide genes system CD/5-Fc+tk/GCV could induce remarkable killing effect on cholangiocarcinoma cells in vitro and transplantation tumors in vivo. The cotransfection of Cx43 gene could enhance the bystander effect and hence the inhibition of carcinoma cells.

Roles of protein kinase C and fructose in hepatic injury caused by obstructive jaundice.

SummaryThe regulating mechanism in hepatic injury caused by obstructive jaundice (OJ) was examined in this study. Rat hepatocytes were harvested byin situ collagenase perfusion and subjected to primary culture. The heptocytes were pre-treated with various concentrations of protein kinase C (PKC) agonist PMA and its inhibitor chelerythrine and cultured for 20 min. After the treatment, 50 μmol/L glycochenodeoxycholate (GCDC) was added and the cells were cultured for an additional 24 h. Cells were then detected by flow cytometry (FCM) and TUNEL. After hepatocytes were treated with different concentrations of fructose and 100 μM GCDC, the cells were examined by FCM and TUNEL. Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct. After BDL, the rats were fed with or without fructos and sacrificed 3, 7, 14 and 21 days after the ligation. The apoptotic status was observed in liver of all rats with TUNEL and PKC protein in liver of OJ was studied by immunohistochemical method. Our results showed that PMA increased GCDC-induced apoptosis and chelerythrine decreased GCDC-induced apoptosis in a concentration-dependent manner. After the treatment with fructose of different concentrations, 100 μM GCDC decreased the apoptotic rate and the apoptotic rate decreased with the increase of fructose concentration. The apoptotic rate of liver was related to the time of OJ. Without the treatment of fructose, PKC and apoptosis index (AI) were highest 14 days after the bile duct ligation. With the treatment of fructose, apoptosis index (AI) and PKC were decreased from the 14th day after the bile duct ligation. It is concluded that PKC is involved in the regulation of apoptosis in the liver cells with OJ and plays important roles in the development and progression of liver injury caused by OJ. Fructose can protect hepatocytes in the bile salt-induced apoptosis by regulating PKC.

DNA content and gene expression in adenoma-carcinoma sequence of gallbladder carcinogenesis

ObjectiveTo study the proliferative change of epithelium and gene expression in adenoma-carcinoma sequence of gallbladder carcinogenesis.MethodsMutations of K-ras gene and APC gene were detected by PCR-RFLP, and the expression of p53 protein was detected by ABC immunohistochemistry. DNA contents were detected by using a TJTY-300 automatic image analyzer.ResultsCell DNA contents and atypia were increased gradually in the adenoma-carcinoma sequence of gallbladder. Aneuploidy in gallbladder adenoma was associated with the size of adenoma. When the diameter of adenoma was more than 1 cm, the aneuploidy was significantly increased (P<0.05). APC gene and K-ras gene mutations were detected in the adenoma-carcinoma pathway. and p53 protein overexpression was not detected in this pathway.ConclusionAdenoma-carcinoma sequence pathway plays an important role in gallbladder carcinogenesis. Gallbladder adenoma has an obvious tendency to malignant transformation, especially in the patients with adenoma of 1 cm in diameter or above. It has molecular abnormalities different from those de novo carcinomas.

Frequency of Loss Expression of DPC4 Protein in Various Locations of Biliary Tract Carcinoma

ObjectiveTo clarify the relationship between loss of expression of DPC4 proteins and pathogenesis of biliary tract carcinoma.Methods71 primary biliary tract carcinomas (BTCa), including 38 common bile duct (CBD) carcinomas, 18 gallbladder carcinomas, and 15 hilar bile ducts (HBD) carcinomas were examined by immunohistochemical staining. In addition, the CBD carcinomas were divided into two groups, a tumor group with metastasis (M+ group, 27 cases) and a tumor group without metastasis (M− group, 11 cases).ResultsThe frequency of loss expression of DPC4 protein was 32.8% in BTCa, 47.3% in CBD carcinoma, 11% in gallbladder carcinoma and 13% in HBD carcinoma. A comparison of the frequency of loss expression of DPC4 showed significantly statistical difference in the CBD carcinoma versus gallbladder carcinoma and HBD carcinoma (P<0.01). The frequency of loss expression of DPC4 was 48.1% in the M+ group and 45.4% in the M− group. There was no significantly statistical difference between them (P>0.05).ConclusionThere is a close relationship between the pathogenesis of BTCa and inactivation of DPC4 with different frequencies of DPC4 gene alteration in various locations of the biliary tract, but inactivation of DPC4 is not related with tumor metastasis in BTCa.

The changes of protein kinase C activity in peripheral blood lymphocytes in the patients with obstructive jaundice and the implication.

SummaryThe roles of protein kinase C (PKC) signal pathway in the pathogenesis of obstructive jaundice were studied. PKC from cytosolic and membrane fractions of peripheral blood lymphocytes (PBL) in 51 patients with obstructive jaundice and 16 cases of normal controls was isolated and purified. The activities of PKC were determined by radioactive isotope γ-32P-ATP-catalyzing assay. The results showed that the total PKC activities in PBL in the patients with obstructive jaundice were significantly increased as compared with those in the normal controls (P< 0.01). Moreover, the membrane PKC activities and their percentages of the total PKC activities were higher in obstructive jaundice group than in those in the normal controls (P< 0.05). The total PKC activities in PBL in the patients with obstructive jaundice were significantly positively correlated with the levels of soluble IL-2 receptor (sIL-2R) (r=0.58,P< 0.01) and the degree of jaundice (T-BIL) (r=0. 67,P< 0.01) in serum. It was concluded that the activities of PKC signal pathway was related with the degree of T-BIL. PKC signal pathway might took part in the activation of T-lymphocytes in the patients with obstructive jaundice and play an important role in the immune regulation and the assessment of pathosis in the patients with obstructive jaundice.