OBJECTIVETo investigate the serum myeloperoxidase (MPO) activity and serum paraoxonase-1 (PON-1) activity in patients with silicosis and observation subjects and their clinical significance.

METHODSSeventy-two patients with silicosis (stage I: 30 cases, stage II: 22 cases, stage III: 20 cases) and 37 observation subjects were selected as a case group, and 110 healthy men were selected as a control group. Serum MPO activity was measured by double-antibody sandwich enzyme-linked immunosorbent assay, and serum PON-1 activity was measured by chemical spectrophotometry.

RESULTSSerum MPO activity was significantly higher in the case group than in the control group [(102.1 ± 15.7) U/L vs. (62.4 ± 11.4) U/L, P < 0.01], but serum PON-1 activity was significantly lower in the case group than in the control group [(85.4 ± 15.7) U/ml vs. (125.4 ± 13.7) U/ml, P < 0.01]. Serum MPO activity was significantly lower in patients with stages I, II, and III silicosis than in the observation subjects [(91.3 ± 13.5) U/L, (85.7 ± 14.4) U/L, and (88.6 ± 14.5) U/L vs. (128.4 ± 16.4) U/L, P < 0.01]. Serum PON-1 activity declined as the stage of silicosis increased; serum PON-1 activity was significantly lower in the patients with stages II and III silicosis than in the observation subjects and the patients with stage I silicosis [(70.4 ± 11.4) U/ml and (67.6 ± 13.7) U/ml vs. (101.5 ± 14.0) U/ml and (89.1 ± 10.1) U/ml, P < 0.01].

CONCLUSIONSerum MPO activity and serum PON-1 activity are valuable for early diagnosis of silicosis and evaluation of patient's condition.

Aged ; Aryldialkylphosphatase ; blood ; Case-Control Studies ; Humans ; Male ; Middle Aged ; Peroxidase ; blood ; Silicosis ; blood

OBJECTIVETo study the effect of Hepatitis B virus X (HBx) gene transfection on expression of human telomerase reverse transcriptase (hTERT) mRNA in human bile duct carcinoma cell lines QBC939 and to elucidate the significance of cis-activation of hTERT mRNA by HBx gene on the carcinogenesis of bile duct.

METHODSQBC939 were cultured in vitro and co-transfected with eukaryotic expression vector containing the HBx coding region and cloning vector containing enhanced green fluorescent protein (EGFP) coding sequence using liposome-mediated gene transduction technique. Thirty six hours after transfection, EGFP expression, the indicator of successful transfection in cells, was determined. Flow cytometry was applied to determine the transfection efficiency. Cells were harvested and total RNA was extracted with TRI(ZOL) Reagent. The expression of hTERT mRNA in QBC939 was assayed by Reverse Transcription Polymerase Chain Reaction. The expression of HBx protein in QBC939 was detected by immunocytochemistry staining and western blotting.

RESULTSThe transfection efficiency was 29.6% for both HBx expression vector and vector control group. The expression of hTERT mRNA was significantly increased when transfected with HBx expression vector than that transfected with OPTI-MEM medium and vector only. The expression of HBx protein could only be found in the cells when transfected with HBx expression vector by immunocytochemistry staining and western blotting.

CONCLUSIONHBx gene transfection may up-regulate the transcriptional expression of hTERT mRNA in bile duct carcinoma cells. The cis-activation of hTERT gene by HBx gene is primary mechanism for carcinogenesis of biliary epithelia after HBV infection.

Bile Duct Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; metabolism ; Trans-Activators ; genetics ; Transfection

OBJECTIVETo transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing cholangiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis.

METHODSQBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVec transfecting technique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry.

RESULTSRT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proliferative index decreased significantly (P < 0.01), the percentage of S phase decreased remarkably (P < 0.05) in antisense vector transfected cells (9.27% +/- 1.91%) compared with that in QBC939 cells without transfection(16.35% +/- 2.87%), and the percentage of G0/G1 phase increased remarkably (P < 0.05) in antisense vector transfected cells (75.16% +/- 4.13%) compared with that in QBC939 cells without transfection (57.31% +/- 10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P > 0.05).

CONCLUSIONTransfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.

Apoptosis ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; Cholangiocarcinoma ; metabolism ; pathology ; Cyclooxygenase 2 ; DNA, Antisense ; genetics ; Humans ; Isoenzymes ; biosynthesis ; genetics ; physiology ; Membrane Proteins ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; physiology ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo investigate the effects of dendritic cells (DCs) transfected with survivin gene, and to observe the effective and specific anti-tumor immunological effect induced by modified DC in vitro.

METHODSSurvivin gene was transfected to DCs with liposomes. Survivin expression could be detected both in DCs cells and in cell culture with method of Western blot. Cytokines as well as cellular surface molecule such as IL-12, TNF-alpha, CD1 alpha, CD83, MHCII, CD80 and CD86 were detected. The competence of inducing human specific cytotoxic T lymphocyte (CTLs) was also detected with MTT.

RESULTSSurvivin expression could be detected both in DCs which were transfected with survivin cDNA and in cell culture superior. The IL-12 and TNF-alpha level was (265.2 +/- 32.7), (437.1 +/- 83.5) pg/ml, and much higher in transgened DC cells than blank DC cells (P < 0.05). CD1 alpha, CD83, MHCII, CD80 and CD86 was high expressed in survivin-DC cells, however, it was low expressed in blank DC cells. The lyse rate to gastric cancer cell, colon cancer cell and bile duct cancer cell was 65%, 77%, and 85% respectively, and these were much higher than those of blank DC cells.

CONCLUSIONSDCs transfected with survivin gene could induce specific cytotoxic T lymphocytes and strikingly raised DC cell's antigen present function, and have specific CTL killing activity.

Antigens, CD ; metabolism ; Dendritic Cells ; immunology ; Gastrointestinal Neoplasms ; therapy ; Humans ; Immunotherapy, Active ; In Vitro Techniques ; Inhibitor of Apoptosis Proteins ; Interleukin-12 ; secretion ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Transfection ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo detect the expression of human telomerase reverse transcriptase (hTERT) protein and mRNA in bile duct carcinomas and the adjacent tissues and to elucidate its role in bile duct carcinogenesis.

METHODSThe expression of hTERT protein and hTERT mRNA in the formalin-fixed paraffin-embedded specimens of 71 cases of bile duct cancers and 39 cases of adjacent tissues was detected by streptavidin-peroxidase immunostaining and in situ hybridization. The correlation was analysed statistically between the expression of hTERT protein and mRNA and clinicopathological parameters bile duct carcinomas.

RESULTSThe positive rate of hTERT protein expression and mRNA expression in malignant specimens was 78.9% (56/71) and 67.6% (48/71), while that in the adjacent tissues was 35.9% (14/39) and 23.1% (9/39), respectively. All the positive signals were found in the hyperplastic biliary epithelia. No significant correlation was established between hTERT expression and clinicopathological parameters.

CONCLUSIONhTERT gene transcription and protein expression is most likely involved in the proliferation and malignant transformation of bile epithelia and the malignant progression of bile duct carcinomas. The detection of hTERT expression may serve elucidating the carcinogenesis of bile duct.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; enzymology ; pathology ; DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Telomerase ; analysis ; genetics

OBJECTIVETo investigate the technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessel or portal vein.

METHODSFrom March 2005 to March 2007, thin slice scan and vessel-reconstruction of 56 patients of malignant tumor in pancreatic head with pressed superior mesenteric blood vessels or portal vein were carried out using multidetector spiral CT to evaluate whether peripheral vessels of pancreatic tumor were invaded and whether the tumor was resectable. During the operation, 3 vascular blocking bands for superior mesenteric vein, portal vein and spleen vein or 4 vascular blocking bands (additional one for inferior mesenteric vein) were preset. Under the cross and traction between superior mesenteric vein and superior mesenteric artery, resected the uncinate process of pancreas thoroughly. Using those methods, radical pancreaticoduodenectomy for 56 patients above-mentioned were successfully accomplished.

RESULTSThe accuracy for preoperative judging by using multidetector spiral CT whether the peripheral vessels of pancreatic cancer were invaded and whether the tumor was resectable was 98% and 100% separately. Thirty-seven of 56 patients, whose superior mesenteric blood vessels or portal veins were pressed by the tumor of pancreatic head, were operated using 3 vascular blocking bands and 2 patients using 4 vascular blocking bands, followed by suturing the bleeding points of the superior mesenteric vein with 5-0 vascular suture Proline. One patient's superior mesenteric vein was partially resected and restored. The operations cost 5-8 h each and the blood loss was 200-600 ml. There were no operative or postoperative hemorrhage or pancreatic juice leakage. According to the follow-up up to now, 2 patients died of multiple live tumor metastases 7 and 9 months separately after operation, the other 54 patients were still alive.

CONCLUSIONSThin slice scan and vessel-reconstruction using multidetector spiral CT can accurately judge whether the blood vessels near the pancreatic tumor were invaded and whether the tumor was resectable, using 3 vascular blocking bands or 4 vascular blocking bands and cross, traction of the superior mesenteric blood vessels, operator can easily accomplish the radical pancreaticoduodenectomy of malignant tumor in pancreatic head with pressed superior mesenteric blood vessels and portal vein, which was not resectable or need combined resection of the blood vessels in the traditional opinion.

Adult ; Aged ; Female ; Humans ; Male ; Mesenteric Artery, Superior ; pathology ; surgery ; Mesenteric Veins ; pathology ; surgery ; Middle Aged ; Neoplasm Invasiveness ; Pancreas ; pathology ; Pancreatic Neoplasms ; pathology ; surgery ; Pancreaticoduodenectomy ; methods ; Portal Vein ; pathology

OBJECTIVETo detect the expression of HBV X gene (HBx mRNA) in extrahepatic biliary tract carcinomas and the adjacent non-cancerous tissues, and to analyzed the relationship between HBV infection and incidence of biliary tract carcinomas, thereby to elucidate the possible role of HBx in the carcinogenesis of biliary tract.

METHODSThe plasmid pSPX46 was digested by appropriate restriction enzyme. HBx fragment was obtained through gel extraction kit. The digoxigenin-labeled DNA probes for HBx mRNA were prepared by a random prime technique. The expression of HBx mRNA was detected in formalin-fixed- paraffin-embedded specimens from 71 cases of biliary tract carcinomas and 39 specimens of non-cancerous tissues adjacent to cancer by in situ hybridization. The correlations between HBx mRNA expression and clinicopathological parameters were statistically analysed in 71 cases of biliary duct carcinomas.

RESULTSForty-three of 71 malignant specimens had detectable HBx mRNA expression with a positive rate being 61%. Only 7 of 39 specimens of non-cancerous tissues adjacent to cancer had weak HBx mRNA expression, with a positive rate being 18%, and all these positive signals were found in the hyperplastic biliary epithelium. No significant correlation was found between HBx mRNA expression and clinicopathological parameters, but a strong positive correlation was found between HBx mRNA and protein expression.

CONCLUSIONThere is a high frequency of HBx mRNA expression in extrahepatic biliary tract carcinomas. HBV infection and its gene integration might play a role to certain extent in the development of biliary tract carcinomas.

Adult ; Aged ; Aged, 80 and over ; Base Sequence ; Bile Ducts, Extrahepatic ; pathology ; Biliary Tract Neoplasms ; complications ; virology ; Female ; Hepatitis B ; complications ; virology ; Hepatitis B virus ; genetics ; Humans ; In Situ Hybridization ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; genetics ; Trans-Activators ; genetics

Objective To investigate the status of genotype of the KPC(Klebsiella pneumoniae carbapenemase)-encoding genes in Pan-resistant K. Pneumoniae, isolated from the 98th Hospital of People' s Liberation Army, Huzhou district, Zhejiang province, China. Methods 19 strains of Pan-resistant K. Pneumoniae were isolated from the inpatients between November, 2008 and July,2009. Phenotypic confirmatory test for suspected carbapenemases production were carried out by Modified Hodge test. Carbapenemase gene of blaKPC was analyzed by PCR and verified by DNA sequencing. Results In 19 strains of K. Pneumoniae, the positive rates of Modified Hodge test and gene of blaKPC were both 100.0%. These genes all belonged to blaKPC-2 subtype confirmed by nucleotide sequence analysis. Among them, the blaKPC-2 gene sequence of the HZ001 strain (its original serial number was HZ9871 ) had been registered in GenBank (GenBank Accession Number: GU086225).Conclusion All of the Pan-resistant K. Pneumoniae isolated from the inpatients harbored blaKPC-2 type carbapenemases gene and causing an outbreak in a hospital. Carbapenemases that producing type KPC-2 might be the major reason which causing the resistance to Carbapenems antibiotics.

OBJECTIVEIn order to study the diagnosis and treatment of HBV and HCV infection.

METHODSWe retrospectively analysed clinical data of 680 patients with cholangiocarcinoma from 1995 to 2001 and stated by SPSS software.

RESULTS(1) The fastigium of cholangiocarcinoma was 60 - 65 years old. The incidence of cholangiocarcinoma was higher in aged males and the sex ratio (male:female) was 1.36:1. (2) The proximal cholangiocarcinoma was most (41.6%) and distant cholangiocarcinoma was secondly (28.7%). (3) Most patients of cholangiocarcinoma were late. The resection rate was low and the rate of radical operation was 21.6% (147/680). (4) The incidence of proximal cholangiocarcinoma was higher in the positive Serologic marks for HBV and HCV and course of diseases was short. Moreover, the pathology of. positive Serologic marks for HBV and HCV trended to low-differentiation and invasion, metastasis and the resection rate was lower.

CONCLUSIONSCholangiocarcinoma is common in the aged males. The infection of HB(C)V and hilar cholangiocarcinoma are correlated and incline to the proximal bile duct. The hilar cholangiocarcinoma infected HB(C)V may have higher malignant degree in biological characteristics and more badly prognosis.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; epidemiology ; etiology ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; epidemiology ; etiology ; Female ; Hepatitis B ; complications ; Hepatitis C ; complications ; Humans ; Incidence ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo study the effect of hepatitis C virus core protein (HCV-C) on human normal biliary epithelial cells (BEC) transformation and tumor development.

METHODSBEC cells were transfected with plasmid pcDNA HCV-C (expressing HCV-C) by lipofectamine and selected in G418. The expression of HCV-C gene and protein was determined by PCR and immunohistochemical staining, respectively. Biological effect of transfected cells was observed through cell proliferation assay, anchor independent growth, and tumor development in nude mice. The expression of HCV-C protein in the induced tumor was evaluated by immunohistochemistry.

RESULTSHCV-C was strongly expressed in BEC cells transfected with plasmid pcDNA HCV-C and the positive signal was located in cytoplasm. The HCV-C expression protein in the induced cytoplasm. Cell proliferation assay showed that the population doubling time in the pcDNA HCV-C transfected cells was much shorter than that in the pcDNA3 and non-transfected cells (14 h, 28 h, 30 h respectively). The cloning efficiencies of transfected cells with pcDNA HCV-C, pcDNA3 and non-transfected cells were 36%, 2.5% and 1.5%, respectively (P < 0.01). Tumor developed in nude mice inoculated with pcDNA HCV-C transfected cells after the inoculation. HE staining showed bile duct carcinoma character and immunohistochemistry confirmed HCV-C expression in the tumor tissue. The positive control group also showed tumor development, while no tumor mass obtained in the nude mice inoculated with pcDNA3 and non-transfected cells even 36 days after the injection.

CONCLUSIONHCV-C protein showed human normal biliary epithelial cells transformation and tumorigenic features.

Animals ; Bile Duct Neoplasms ; etiology ; Bile Ducts ; cytology ; Cell Transformation, Neoplastic ; Cell Transformation, Viral ; Cells, Cultured ; Epithelial Cells ; pathology ; Female ; Hepacivirus ; Humans ; Mice ; Mice, Nude ; Plasmids ; Transfection ; Viral Core Proteins ; physiology