Aged ; Case-Control Studies ; Humans ; Male ; Malondialdehyde ; metabolism ; Middle Aged ; Oxidation-Reduction ; Serum ; metabolism ; Silicosis ; blood ; Superoxide Dismutase ; metabolism

OBJECTIVEThe aim of this study was to explore the possibility of creating a toxin, C-CPE-ETA', by fusing C-terminal high affinity binding domain of CPE (C-CPE) with a truncated form of Pseudomonas aeruginosa exotoxin A (ETA') and to examine whether C-CPE-ETA' could specifically target CLDN-3, 4 molecule and the targeted toxin was cytotoxic against CLDN-3,4-overexpressing ovarian cancer.

METHODSCLDN-3 and CLDN-4 expressions were analyzed at the mRNA level in three ovarian cancer cell lines and epithelial ovarian cancer tissues from 20 patients. After transforming an expression plasmid of C-CPE-ETA' into E. coli BL21 (DE3) plysS strain, the recombinant protein was purified using His-Bind resin chromatography column and analyzed by Western blot and Coomassie blue staining. The specific binding, proapoptotic and cytolytic activities were evaluated by flow cytometry, fluorescence microscopy with the JC-1 probe and MTT assay in CLDN-3,4-overexpressing ovarian cancer cells.

RESULTSQuantitive RT-PCR results showed there existed high levels of CLDN-3 and CLDN-4 in ovarian cancer cells, CAOV3, OVCAR3 and SKOV3. Moreover, high expressions of CLDN-3 and CLDN-4 were observed in 90.0% (18/20) and 60.0% (12/20) of ovarian cancer tissues, with an expression level 10-fold higher than that in the normal ovarian tissue. A 58 000 recombinant protein C-CPE-ETA' was demonstrated by Western blot and Coomassie blue staining. Purified and recombinant C-CPE-ETA' was bound with high affinity to CLDN-3,4-overexpressing ovarian cancer cells, CAOV3, OVCAR3 and SKOV3 cells. C-CPE-ETA' was strongly proapoptotic and cytotoxic towards the CLDN-3,4-overexpressing ovarian cancer cells. The concentration of IC(50) was 7.364 ng/ml for CAOV3 cells, 8.110 ng/ml for OVCAR3 cells and 22.340 ng/ml for SKOV3 cells, respectively. However, control CLDN-3,4-deficient cell line HUVEC was not susceptible to the recombinant C-CPE-ETA' at a concentration up to 10 µg/ml.

CONCLUSIONSThe C-CPE-ETA' protein exhibits remarkably specific cytotoxicity for CLDN-3,4-overexpressing ovarian cancer cells. Its therapeutic potential warrants further development for ovarian cancer molecular targeted therapy.

ADP Ribose Transferases ; metabolism ; physiology ; Apoptosis ; Bacterial Toxins ; metabolism ; Cell Line, Tumor ; Claudin-3 ; Claudin-4 ; Claudins ; genetics ; metabolism ; Enterotoxins ; metabolism ; physiology ; Exotoxins ; metabolism ; physiology ; Female ; Humans ; Immunotoxins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Recombinant Fusion Proteins ; metabolism ; physiology ; Virulence Factors ; metabolism ; physiology

Objective To explore the effect of long non-coding RNA(lncRNA)SOCS2-AS1 on the proliferation,invasion and migration of gastric cancer cells and its potential mechanism.Methods 40 cases of gastric cancer tissues and 40 cases of adjacent tissues were selected as experimental specimens.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of lncRNA SOCS2-AS1 in gastric cancer tissues and adjacent tissues.Human gastric cancer SGC-7901 cells were divided into vector group,pcDNA-SOCS2-AS1 group,NC-siRNA group,SOCS2-AS1-siRNA group,pcDNA-SOCS2-AS1+vector group and pcDNA-SOCS2-AS1+pcDNA-YAP1 group.Vector group,pcDNA-SOCS2-AS1 group,NC-siRNA group and SOCS2-AS1 siRNA group were transfected with vector,pcDNA-SOCS2-AS1,NC-siRNA and SOCS2-AS 1 siRNA plasmids,respectively;the pcDNA-SOCS2-AS1+vector group were transfected with the pcDNA-SOCS2-AS1+vector plasmids;the pcDNA-SOCS2-AS1+pcDNA-YAP1 group were transfected with both pcDNA-SOCS2-AS1+pcDNA-YAP1 plasmids simultaneously.RT-qPCR was used to detect the expression level of lncRNA SOCS2-AS1 in cells;Western blotting was used to detect the expression of Hippo-YAP pathway-related proteins LATS1 and YAP1 in gastric cancer cells;EDU staining was used to detect the proliferation of gastric cancer cells;Transwell assay was used to detect cell migration and invasion.Results The relative expression levels of SOCS2-AS1 in gastric cancer tissues,adjacent tissues,normal gastric epithelial cells GES-1 and gastric cancer cells SGC-7901 were 1.00±0.18,0.38±0.09,1.03±0.21 and 0.42±0.11.Compared with the adjacent tissues and normal gastric epithelial cell line GES-1,SOCS2-AS1 was significantly down-regulated in gastric cancer tissues and gastric cancer cell line SGC-7901,and the differences were statistically significant(P＜0.05).The cell proliferation rates in the vector group,pcDNA-SOCS2-AS1 group,NC-siRNA group and SOCS2-AS1-siRNA group were(36.23±2.53)％,(18.15±1.04)％,(35.83±2.15)％and(56.11±6.57)％;the invasive cell numbers were 168.15±19.11,54.36±6.27,124.47±14.53 and 238.62±21.34;the numbers of migrating cells were 194.22±16.33,91.61±8.47,148.14±16.25 and 279.31±24.68;the relative expression levels of LATS1 protein were 0.93±0.08,2.86±0.21,0.96±0.09 and 0.31±0.03;the relative expression levels of YAP1 protein were 0.95±0.11,0.28±0.04,0.92±0.12 and 3.16±0.28;the pcDNA-SOCS2-AS1 group was significantly lower than the vector group,and the SOCS2-AS1-siRNA group was significantly higher than the NC-siRNA group,and the differences were statistically significant(all P＜0.05).The cell proliferation rates of pcDNA-SOCS2-AS1+vector group and pcDNA-SOCS2-AS1+pcDNA-YAP1 group were(33.62±5.73)％and(88.46±9.17)％;the number of invasive cells were 53.61±6.44 and 131.73±15.29;the number of migrating cells were 68.35±7.63 and 159.81±16.47,the differences were statistically significant(all P＜0.05).Conclusion LncRNA SOCS2-AS1 inhibits the proliferation,invasion and migration of gastric cancer cells by activating the Hippo-YAP signaling pathway.

OBJECTIVETo study the prevalence of hepatitis B virus (HBV) genotype in 5 cities of Fujian province and the clinical implications of distinct genotypes in HBV-related liver diseases.

METHODSHBV genotype was determined by the restriction fragment length polymorphism analysis in patients with chronic HBV infection in 5 cities of Fujian province. The relationship between HBV genotype and its clinical implications was studied by multinomal logistic regression and correspondence analysis.

RESULTSOf the 431 HBV DNA positive patients detected by PCR, 275 (63.8%) belonged to HBV genotype B, 100 (23.2%) to genotype C, 51 (11.8%) to genotype D and D-mixed genotype. Genotype A, E and F were not found. Multinomal logistic regression showed that genotype B was more prevalent in Quanzhou and Sanming cities than in Fuzhou (P = 0.002, P = 0.006), and genotype B appeared significantly more common in asymptomatic carriers (ASC), chronic hepatitis B (CHB) and severe hepatitis (SH). Genotype C was most prevalent in patients with liver cirrhosis (LC) (47.0%) than in those with ASC (14.5%) and SH (14.7%) (P = 0.009, P < 0.001). The positive rate of hepatitis B e antigen was higher in patients with genotype C than in those with genotype B and genotype D (56.0% vs. 52.4%, P = 0.008, and 56.0% vs. 30.8%, P = 0.051, respectively). By correspondence analysis, genotype D and D-mixed genotype seemed to be correlated with hepatocellular carcinoma (HCC).

CONCLUSIONS(1) The major popular genotypes of HBV were B, C and D in Fujian. (2) Data of our study suggested that the geographic distribution of genotype B and C might be different in some cities of Fujian. (3) Genotype B might have a tendency to lead to SH in younger patients with chronic hepatitis B and the development of LC might be associated with genotype C among the elder patients. (4) Genotype D appeared to associate with development of HCC, which called for further study to confirm.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; Female ; Gene Frequency ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; genetics ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

OBJECTIVETo investigate whether there are different stromal compositions in the prostate tissue of patients with benign prostatic hyperplasia (BPH) and evaluate their significance in the course of the disease.

METHODSForty-three surgical or bioptic prostatic specimens of BPH and 5 autoptic normal prostatic specimens were stained by the Masson method to display the elements of the muscle fiber and collagen. The relationship of the changes in the prostatic stromal composition was analyzed with the degree of bladder outlet obstruction (BOO) , IPSS and medication results.

RESULTSThe mean ratio of muscle fiber to collagen in the normal prostate tissue was (3.2 +/- 0.2):1, significantly higher than that of the BPH patients (1: [4.7 +/- 3.1] ) (P < 0.01); that in the BPH patients with BOO was 1: (5.4 +/- 3.7) markedly lower than in those without BOO (1: [2.5 +/- 1.1] ) (P = 0.02); that in the BPH patients with severe prostatic symptoms was 1: (9.1 +/- 2.9), remarkably lower than in those with moderate (1: [5.3 +/- 3.4]) and mild prostatic symptoms (1: [2.8 +/- 1.7]) (P < 0.01); and that in the BPH patients with satisfactory medicinal therapeutic results was 1:(2.3 +/- 1.9), significantly higher than in those with poor therapeutic results (1: [7.6 +/- 4.3]) (P < 0.01).

CONCLUSIONThe stromal composition in the prostatic tissue of BPH patients undergoes different degrees of changes. More obvious BPH symptoms and poorer therapeutic results are associated with a bigger proportion of collagens and a smaller proportion of muscle fibers in the prostatic tissue. These changes may play an important role in the development and progression of BPH.

Aged ; Aged, 80 and over ; Case-Control Studies ; Fibrosis ; Humans ; Male ; Middle Aged ; Prostate ; pathology ; Prostatic Hyperplasia ; pathology ; Urinary Bladder Neck Obstruction ; pathology

BACKGROUNDThe adverse health effects of lead for children under 6 years are well known. Studies to assess the lead exposure among children in China are small in sample size and lack of national representative data. The aim of this study therefore was to describe blood lead levels and identify risk factors for lead exposure among children aged 0 to 6 years living in 16 cities in China.

METHODSWe analyzed data from blood lead levels surveillance in China carried out in 16 large cities between 2004 and 2008. A stratified clustered random sampling strategy was used. A total of 69 968 children aged 0 to 6 years were included. We conducted multiple Logistic regression analyses to explore risk factors to high blood lead level.

RESULTSThe geometric mean blood lead level of the children was 4.50 µg/dl (median: 4.90 µg/dl; IQR: 3.20 - 7.00 µg/dl). Overall prevalence of blood lead level ≥ 10.00 µg/dl among 0- to 6-year-old children was 7.57%. But the proportion of blood lead level ≥ 5.00 but < 10.00 µg/dl was 42.12%. Blood lead levels were significantly higher in boys (4.63 µg/dl) than in girls (4.35 µg/dl) (P < 0.0001). The geometric mean blood lead levels and prevalence of blood lead level ≥ 10.00 µg/dl increased with age (P < 0.0001 for the two trends). After controlling for sociodemographic, dietary and behavior factors, multivariable analysis indicated that lower maternal education, male gender, younger age, often biting pencil or/and toys, walking or playing for long time on the street, not washing hands before eating are major risk factors for higher lead levels.

CONCLUSIONSThe blood lead levels among Chinese children in urban areas are lower than previous studies but close to those of developed countries. However, children with low lead exposure account for almost half and the sociodemographic factors (age, male sex, and low mother education level) continue to be associated with higher blood lead levels.

Child, Preschool ; China ; Female ; Humans ; Infant ; Infant, Newborn ; Lead ; blood ; Logistic Models ; Male ; Time Factors ; Urban Health

BACKGROUNDComputed tomography (CT)-guided transthoracic lung biopsy is a well-established technique for the diagnosis of pulmonary lesions. The objective of this study was to evaluate the diagnostic efficiency and complication rate of CT-guided lung biopsy in a Chinese population.

METHODSCT-guided cutting needle lung biopsies were performed in our institution on 1014 patients between January 2000 and October 2010. A chest radiograph was taken after the biopsy. Data about basic patient information, final diagnosis, and complications secondary to biopsy procedure (pneumothorax and bleeding) were extracted.

RESULTSThe diagnostic efficiency of CT-guided lung biopsy was 94.8%; only 53 patients did not get a final diagnosis from lung biopsy. Final diagnoses found 639 malignant lesions (63.0%) and 322 benign lesions (31.8%). Pneumothorax occurred in 131 patients and 15 required insertion of an intercostal drain. Small hemoptysis occurred in 41 patients and mild parenchymal hemorrhage occurred in 16 patients. The overall complication rate was 18.5%.

CONCLUSIONSCT-guided cutting needle biopsy of pulmonary lesions is a relatively safe technique with a high diagnostic accuracy. It can be safely performed in clinical trials.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Needle ; methods ; Female ; Humans ; Lung ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; methods ; Young Adult

OBJECTIVETo examine the expression patterns of short palate, lung and nasal epithelium clone 1 (SPLUNC1) gene in human tissues.

METHODSIn situ hybridization was used to detect the expression of SPLUNC1 gene in 37 different human tissues.

RESULTSWe found that SPLUNC1 gene was not expressed in squamous epithelial cells of the palate, epidermis, esophagus, or the esophagus-cardia junction, metaplastic squamous cells in the nasopharynx, trachea, or uterus cervix, or tumor cells of esophageal squamous cell carcinoma or lung squamous cell carcinoma. SPLUNC1 gene was not expressed in the single layer columnar epithelia cells in the stomach, gallbladder, jejunum, colon, endometrium, or uterus cervix. SPLUNC1 expression was detected mainly in pseudostratified columnar epithelial cells in the nasopharynx, trachea and bronchi, and was gradually down-regulated from the upper to lower end of the respiratory tract, but was not detected in the lung tissues. SPLUNC1 expression was detected not only in the duct and serous gland cells in the parotid and submandibular glands, but also in cells of submucosal serous glands in the nasopharynx and lung, but not in the cells of the mucosal glands. The parietal cells of the gastric submucosa and epithelial cells of the lobula and ducts of the mammary glands expressed SPLUNC1. The adenocarcinoma cells in the lung, stomach, colon, mammary gland, uterus endometrium and cervix showed strong expressions of SPLUNC1 gene.

CONCLUSIONSPLUNC1 expression is highly cell-specific in association with the cell functions.

Epithelial Cells ; metabolism ; Gene Expression ; Glycoproteins ; genetics ; metabolism ; Humans ; Organ Specificity ; Phosphoproteins ; genetics ; metabolism

BACKGROUNDCoronary artery disease (CAD) is generally considered as a disease of middle-aged men. It is widely accepted that the risk for CAD of premenopausal women is low because of hormone protection. Based on our clinical experience, more and more premenopausal women suffer from angina and myocardial infarction without adequate concern. Even now, there are still limited detailed data to describe the characteristics, mechanism and prognosis of premenopausal CAD patients. This article aimed to analyze the clinical and angiographic characteristics of premenopausal women with CAD.

METHODSA total of 565 premenopausal women and 721 postmenopausal women (56 - 60 years old) who underwent coronary angiography for the first time from April 2004 to December 2007 were enrolled. The clinical data and coronary angiographic characteristics (presence, localization, length and severity) were compared between the premenopausal and postmenopausal CAD groups.

RESULTSPremenopausal CAD patients presented less frequently with hypertension, diabetes mellitus and dyslipidemia compared with postmenopausal CAD patients (55.0% vs 66.0%, 15.0% vs 31.5%, 23.9% vs 37.4%, respectively; all P < 0.05). Although we found more frequent involvement of single vessel in premenopausal CAD (43.2% vs 26.9%, P = 0), and triple vessels in postmenopausal (56 - 60 years old) CAD patients (33.8% vs 20.4%, P = 0), much more severe lesions (> or = 90%) at left main (2.9% vs 1.1%, P = 0.048) and proximal left anterior descending artery (LAD) (28.2% vs 16.6%, P = 0) in the premenopausal CAD group were found.

CONCLUSIONPremenopausal women with chest discomfort are always found to have obvious atherosclerosis, more inclined to be located at the left main and proximal LAD, which is a strong predictor of an adverse clinical outcome.

Coronary Angiography ; Coronary Artery Disease ; diagnostic imaging ; pathology ; Diabetes Mellitus ; pathology ; Dyslipidemias ; pathology ; Female ; Humans ; Hypertension ; pathology ; Middle Aged ; Postmenopause ; Premenopause