OBJECTIVE: Over-expression of thrombin in ovarian cancer cells is associated with poor prognosis. In this study, we investigated the role of thrombin in inducing epithelial-mesenchymal transition (EMT) in SKOV3 epithelial ovarian cancer cells. METHODS: After thrombin treatment SKOV3 cells were subjected to western blots, reverse-transcription PCR, and enzyme-linked immunosorbent assay to quantify EMT-related proteins, mRNA expression of SMAD2, DKK1, and sFRP1, and the secretion of matrix metalloproteinases (MMPs) and cytokines. Meanwhile, invasion ability was evaluated using transwell assays. RESULTS: The results indicated a dose- and time-dependent down-regulation of E-cadherin and upregulation of N-cadherin and vimentin in thrombin-treated SKOV3 cells, compared with the thrombin-free control group (p<0.05). There was a dose- and time-dependent increase in the levels of SMAD2 and DKK1 mRNAs and a decrease in the levels of sFRP1 mRNA in thrombin-treated SKOV3 cells compared to control cells (p<0.05). Thrombin-treated SKOV3 cells exhibited increased secretion of MMP-9, MMP-2, interleukin (IL)-8, and IL-6 and increased invasion compared to untreated cells (p<0.05). Thrombin altered the morphology of SKOV3 cells to a spindle-like phenotype. Addition of hirudin to thrombin-treated cells reversed the effects of thrombin. CONCLUSION: Thrombin induced EMT and promoted the invasion of SKOV3 cells, possibly via distinct signaling pathways. Hirudin inhibited the effects of thrombin, suggesting that anticoagulant therapy could be a novel therapeutic strategy for ovarian carcinoma.
Blotting, Western ; Cadherins ; Cytokines ; Down-Regulation ; Enzyme-Linked Immunosorbent Assay ; Epithelial-Mesenchymal Transition ; Hirudins ; Interleukin-6 ; Interleukins ; Matrix Metalloproteinases ; Neoplasms, Glandular and Epithelial ; Ovarian Neoplasms ; Phenotype ; Polymerase Chain Reaction ; Prognosis ; Proteins ; RNA, Messenger ; Thrombin ; Up-Regulation ; Vimentin

OBJECTIVEIn order to provide pathologic reference for therapeutic rationale, the pathological changes of the pulmonary vasculature in patients with pulmonary atresia with ventricular defect and patent ductus arteriosus were observed by contrast with normal control group.

METHODSLung biopsies were taken in the operation in 10 children suffered from pulmonary atresia with ventricular septal defect associated with patent ductus arteriosus (PA group). Autopsy specimens were obtained from 10 children died of non-cardiovascular diseases as normal control group. The tissue was fixed with buffered formalin, routinely prepared by impregnated in wax. Sections were stained by hematoxylin-eosin, Weigert's elastic stain counter-stained by van Gieoson's method. Seven parameters were obtained including percentage of media thickness (MT%), percentage of media section area (MS%), numbers of vascular per square centimeter (VPSC), mean alveolar number (MAN), mean linear intercept (MLI), proportion of parenchyma area in total area (PPA), and alveolar/vascular ratio per unit area (AVR) by a computer image processor by quantitative analysis.

RESULTSThere were significant difference between the two groups in MAN, VPSC, and AVR (P < 0.05). VPSC was significantly lower in PA group than in control group (P < 0.01). Other parameters had no significant difference. The mean alveolar diameter had an increased trend in PA group, although there was no significant difference. MS% of nearly 50% patients was closed to the normal value in PA group. The shape of pulmonary arteriole was irregular. There were few muscular arteries in a field of vision.

CONCLUSIONSThe density of muscular arteries decreases in patients with pulmonary atresia with ventricular septal defect and patent ductus arteriosus, but percentage of media thickness and percentage of media section area of pulmonary arterioles are close to the normal value. Diminished flow in pulmonary circulation has a significant effect on numbers of pulmonary arterioles per square centimeter that impact the selection of surgical method and the effect of operation because of the reduction pulmonary arterioles. The decrease of mean alveolar number results in compensatory enlargement of alveolar diameter. The impaired lung development is a major cause of abnormal lung function. Feasible and earlier operation, which can increase pulmonary flow and promote development of pulmonary vasculature will be helpful to restore lung function.

Abnormalities, Multiple ; pathology ; Child, Preschool ; Ductus Arteriosus, Patent ; pathology ; Female ; Heart Septal Defects, Ventricular ; pathology ; Humans ; Infant ; Male ; Pulmonary Artery ; pathology ; Pulmonary Atresia ; pathology

OBJECTIVETo analyse the relationship between the quantitative structural study of lung and right ventricle outflow tract reconstruction in infants with tetralogy of Fallot.

METHODSLung biopsies were taken during the operations in 16 infants suffered from tetralogy of Fallot. Autopsy specimens were obtained from 5 infants died of non-cardiovascular diseases as normal control group. All patients underwent one staged repair. The techniques of right ventricular outflow tract reconstruction included pulmonary valve commissurotomy (n = 3), transanular pericardial patch (n = 4), and transannular homologous monocuspid valve patch (n = 8); homograft was used in one patient because of the abnormal coronary artery. The diameters of main pulmonary artery (MPA), left pulmonary artery (LPA), and right pulmonary artery (RPA) were measured during operation. The tissue was fixed with buffered formalin and routinely impregnated in wax. Sections were stained by hematoxylin-eosin, and Weigert's elastic stain counter-stained by van Gieoson's method. Seven parameters of the small pulmonary arteries were obtained, including percentage of media thickness (% MT), percentage of media section area (% MS), numbers of pulmonary small artery per square centimeter (APSC), mean alveolar number (MAN), mean linear intercept (MLI), proportion of parenchyma area in total area (% PPA), and alveolar/ small arterial ratio per unit area (AAR) by a computer-based image processor for quantitative analysis.

RESULTSIn the TOF group, % MT, % MS, and APSC significantly decreased, while MLI and AAR significantly increased (P < 0.05, compared with the control group). APSC decreased in turn after separately using three different techniques of right ventricular outflow tract reconstruction (i. e. pulmonary valve commissurotomy, transannular pericardium patch, and transannular homologous monocuspid valve patch), which was paralleled with the diameters of MPA, LPA, and RPA. RPA correlated with APSC (r = 0.754, P = 0.001).

CONCLUSIONSThe development of pulmonary small arteries and alveoli are directly affected by the diminished pulmonary flow in infants with tetralogy of Fallot. Right ventricle outflow tract reconstruction may be indicated according to the developmental degree of central pulmonary artery.

Biopsy, Needle ; Child, Preschool ; Heart Ventricles ; surgery ; Humans ; Infant ; Lung ; pathology ; Reconstructive Surgical Procedures ; methods ; Tetralogy of Fallot ; pathology ; surgery

Objective The aim is to investigate the correlation between bisphenol A (BPA) exposure and tumor tissue ceramide (Cer) as well as serum tumor markers in colorectal cancer (CRC). Methods The morning urine and CRC tumor tissue were collected from 84 patients with CRC. The concentration of urine BPA was determined by liquid chromatography-mass spectrometer (LC-MS), urine BPA concentration was corrected with creatinine (Cr). Cer concentration of CRC tumor tissue was detected by Enzyme-linked immunosorbent assay (ELISA). The correlations of urine BPAcr, Cer content of CRC tumor tissue and tumor markers were analyzed. Results Cer content in CRC tumor tissue was positively correlated with BPAcr (r=0.784, P＜0.001). Regression analysis showed that the regression coefficient of Cer content in CRC tumor tissue and BPAcr was 0.218 (95% CI: 0.18-0.26), which was statistically significant (P＜0.001). There were significantly differences in CRC tumor tissue Cer and urine BPAcr between the CEA positive and negative groups, CA125 positive and negative groups, and CA19-9 positive and negative groups (all P<0.05), while there was no significant difference between AFP positive and negative groups in CRC tumor tissue Cer and urine BPAcr (P=0.247). Serum CEA, CA125 and CA19-9 were positively correlated with urine BPAcr (r values were 0.348, 0.251, 0.281, respectively, all P＜0.05) and Cer content in CRC tumor tissue (r values were 0.265, 0.309, 0.263, respectively, all P＜0.05). Conclusions BPA exposure may cause an increase of Cer in CRC tumor tissue and abnormalities in serum tumor markers, suggesting that BPA exposure may participate in the development and occurance of CRC by affecting the metabolism of Cer in CRC tumor tissue.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; HIV Infections ; physiopathology ; Humans ; Liver Diseases ; physiopathology ; virology ; Male ; Middle Aged ; Risk Factors ; Young Adult

OBJECTIVETo explore the existence of vasculogenic mimicry (VM) in ovarian carcinoma and its correlationship with the clinicopathologic features and prognosis of the tumor.

METHODSA total of 84 ovarian carcinoma cases were collected with complete clinical and prognostic data. CD31 immunohistochemistry and PAS special stain were used to investigate VM in the tumor tissue. Immunohistochemical staining of VEGF, MMP-2, MMP-9, E-cadherin, beta-catenin, and Vimentin were used to explore the pathogenesis of VM.

RESULTSTotally 36 of 84 cases exhibited evidence of VM. FIGO classification, pathologic grades and histological types were significantly different between the VM and non-VM groups. Expression of VEGF, MMP-2, MMP-9, E-cadherin and beta-catenin were higher in the VM group than in the non-VM group. Kaplan-Meier survival curve analysis showed that cases of the VM group had a lower survival rate than that of the non-VM group (P = 0.04).

CONCLUSIONSVasculogenic mimicry exists in ovarian carcinoma. Ovarian carcinomas with a high grade malignancy have a high incidence of VM formation, a higher incidence of metastases and a lower survival rate. High expression of MMP-2 and MMP-9 may contribute to the formation of VM in the ovarian cancer.

Cadherins ; metabolism ; Carcinoma, Endometrioid ; metabolism ; pathology ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Female ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neovascularization, Pathologic ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism ; beta Catenin ; metabolism

OBJECTIVEThe development of pulmonary vascular bed is strongly flow-dependent. Abnormal pulmonary blood flow leads to pulmonary pathological changes. This study aimed to observe the pathological changes of small pulmonary arteries and alveoli in complex congenital heart defect with diminished pulmonary blood flow but without aortopulmonary collateral artery (APCA) and patent ductus arteriosus (PDA) in infants and young children.

METHODSAutopsy pulmonary specimens obtained from 5 children who died of non-cardiovascular diseases were used as the control group (age: 4-18 months). Fifty-six children (age: 4-36 months) with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA served as the study group, including 34 cases of tetralogy of Fallot, 7 cases of double outlet right ventricle with pulmonary stenosis, 9 cases of single ventricle with pulmonary stenosis, 4 cases of tricuspid atresia with pulmonary stenosis and 2 cases of complete atrioventricular septal defect with pulmonary stenosis. Pulmonary specimen sections were stained by hematoxylin-eosin and Weigert-Van Gieson. Percentage of media thickness (MT%), percentage of media section area (MS%), number of small arterial per square centimeter (APSC), mean alveolar number (MAN), mean linear intercept (MLI), proportion of parenchyma area in total area (PPA%) and alveolar to small arterial ratio per unit area (AAR) were measured by morphologic quantitative analysis.

RESULTSMT% (10.93+/-2.87% vs 15.08+/-2.51%), MS% (18.97+/-5.56% vs 25.04+/-3.87%) and APSC (202.43+/-67.45 vs 441.69+/-65.29) decreased significantly in the study group compared with the control group (P<0.01). The internal diameter of small pulmonary artery (80.26+/-21.57 microm vs 58.53+/-10.29 microm; P<0.05), AAR (46.59+/-14.43 vs 34.46+/-4.98; P<0.01) and MLI (144.98+/-44.87 microm vs 108.39+/-20.76 microm; P<0.05) increased significantly compared with the control group.

CONCLUSIONSThe media of small pulmonary arteries becomes thinner, the lumen of small pulmonary arteries becomes larger, and the number of small arterial per square centimeter and the mean alveolar number are reduced in infants and young children with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA.

Aorta ; abnormalities ; Child, Preschool ; Collateral Circulation ; Ductus Arteriosus, Patent ; pathology ; Female ; Heart Defects, Congenital ; pathology ; Humans ; Infant ; Lung ; pathology ; Male ; Pulmonary Artery ; abnormalities ; Pulmonary Circulation

AIM:To investigate the inhibitory effect of allicin on apoptosis and caspase-12 activation of macro-phage-derived foam cells,and to elucidate the underlying molecular mechanisms. METHODS:RAW264.7 macrophages were pretreated with allicin (12.5,25 and 50 mg/L) or 4-phenylbutyric acid(PBA,4 mmol/L) for 1 h and then treated with oxidized low-density lipoprotein(ox-LDL,100 mg/L) or tunicamycin(TM,4 mg/L) for 24 h. The cell viability and apoptosis were examined by MTT assay and flow cytometry with Annexin V-FITC/PI staining,respectively. The activities of caspase-3 in the cells and lactic dehydrogenase (LDH) in the medium were measured. The protein levels of caspase-12 were determined by Western blot. The intracellular lipid accumulation was measured with oil red O staining and the content of intracellular total cholesterol was determined by enzymatic colorimetry. RESULTS:Similar to the endoplasmic reticulum stress (ERS) inhibitor PBA, allicin inhibited ox-LDL-induced injury of RAW264.7 macrophages in a concentration-de-pendent manner,as determined by the increased cell viability and the decreased LDH leakage,apoptosis and caspase-3 ac-tivity. The decrease in cell viability and increases in LDH leakage and apoptosis induced by TM (an ERS inducer) were also suppressed by allicin. Moreover, similar to PBA, allicin remarkably inhibited ox-LDL- or TM-induced activation of caspase-12. Furthermore, allicin remarkably attenuated ox-LDL-induced lipid accumulation in the RAW264.7 cells and foam cells formation in a concentration-dependent manner. CONCLUSION:Allicin may inhibit macrophage-derived foam cell apoptosis induced by ox-LDL,and the mechanism is partially related to suppressing the activation of caspase-12.

Secretory IgA (SIgA) antibodies in external secretions play an important role in mucosal immune response. Polymeric SIgA was advantageous over monomeric IgA (mIgA) and IgG in several aspects. To express secretory IgA antibody against H5N1 virus, we constructed the secretory component and immunoglobulin J expressing plasmids and co-transfected the plasmids into the Chinese hamster ovary cells (CHO) stably expressing immunoglobulin A. Then we used Zeocin to select the positive clone cells, monoclonal cells stably secreting SIgA was screened through fold dilution method at last. The SIgA antibody secreted from the CHO cells was confirmed by Western blotting, which demonstrated that we had got the complete SIgA molecular. The successful expression of this polymeric anti-H5N1 SIgA in CHO cells will contribute to the production of recombinant SIgA as a preventive agent for infectious disease control.
Animals ; Antibodies, Viral ; biosynthesis ; genetics ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Genetic Vectors ; Immunoglobulin A ; immunology ; Immunoglobulin A, Secretory ; biosynthesis ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

BACKGROUNDLiver injury is one of the most important adverse effects of antiretroviral therapy, leading to therapy changing or discontinuation. Data on liver injury in human immunodeficiency virus-1-infected patients receiving antiretroviral therapy are limited in China. The purpose of this study was to investigate the features of liver injury in human immunodeficiency virus type 1-infected patients receiving non-nucleosides reverse transcriptase inhibitors-based antiretroviral therapy in China.

METHODSSeventy-five patients on antiretroviral therapy containing non-nucleosides reverse transcriptase inhibitors were retrospectively studied. The patients were divided into 2 groups: group 1 (with liver injury, n = 45) and group 2 (without liver injury, n = 30). The features of liver injury were analyzed. The sex, age, baseline CD4 counts, hepatitis B virus (HBV) and/or hepatitis C virus (HCV) co-infection, hepatotoxic drug use and nevirapine or efavirenz use were compared between two groups.

RESULTSForty-five patients (60.0%), 31 (68.9%) males and 14 (31.1%) females, aged 12 to 52 years (averaged (39 ± 9) years), experienced at least one episode of liver injury. Forty (53.3%) patients were co-infected with HBV and/or HCV, 42 (56%) patients had concomitant use of antituberculosis drugs or cotrimoxazole, 46 (61.3%) and 29 (38.7%) patients received regimen containing nevirapine and efavirenz, respectively. Grade 1 liver injuries were observed in 26 (57.8%) patients, grade 2 in 16 (35.6%), grade 3 in 2 (4.0%) and grade 4 in 1 (2.2%). Three (6.7%) patients discontinued highly active antiretroviral therapy (HAART) due to liver injury. In group 1, there were 29 (64.4%) patients co-infected with HBV and/or HCV, 32 (71.1%) patients received regimen containing nevirapine, and 30 (66.7%) patients had concomitant use of anti-tuberculosis drugs or cotrimoxazole, respectively, significantly higher than those in group 2 (11 (36.7%), 14 (46.7%) and 12 (40%), respectively; P = 0.018, 0.033, 0.023, respectively). The sex, age, baseline CD4 counts and disease stage were not factors associated with liver injury.

CONCLUSIONSLiver injury associated with HAART containing non-nucleosides reverse transcriptase inhibitors was mild to moderate and those who were co-infected with HBV and/or HCV, had concomitant use of antituberculosis drugs or cotrimoxazole and received a regimen containing nevirapine were prone to liver injury while receiving HAART.

Acquired Immunodeficiency Syndrome ; drug therapy ; Adolescent ; Adult ; Antiretroviral Therapy, Highly Active ; adverse effects ; Chemical and Drug Induced Liver Injury ; etiology ; Child ; Female ; HIV-1 ; Humans ; Male ; Middle Aged ; Nevirapine ; adverse effects ; Retrospective Studies ; Reverse Transcriptase Inhibitors ; adverse effects