Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo investigate the changes in the proliferation and migration of human ovarian cancer cells (CAOV-3) after knocking down COX-2 gene by RNA interference.

METHODSThe recombinant plasmid of pGenesil-1-siRNA-COX-2 was constructed and transfected into CAOV-3 cells. The transcription of COX-2 gene was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), and the protein expression of COX-2 was determined by Western blotting . MTT assay was used to investigate the proliferation of the transfected CAOV-3 cells, and the cell migration was evaluated using a transwell migration assay.

RESULTSCOX-2 mRNA and protein levels were significantly reduced after pGenesil-1-siRNA-COX-2 transfection into CAOV-3 cells, which showed obvious reduction in the cell proliferation and migration.

CONCLUSIONRNA interference allows obvious COX-2 gene knocking down in human ovarian cancer cells to result in lowered cell growth rate and migration ability. COX-2 gene may become a new therapeutic target for ovarian cancer.

Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclooxygenase 2 ; genetics ; Female ; Humans ; Ovarian Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.
Animals ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Cell Nucleus ; metabolism ; Cerebral Cortex ; metabolism ; Female ; Fibroblast Growth Factor 1 ; administration & dosage ; pharmacokinetics ; Gene Products, tat ; administration & dosage ; pharmacokinetics ; Hippocampus ; metabolism ; Injections, Intravenous ; Male ; Mice ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; administration & dosage ; pharmacokinetics

Objective:To explore the relationship among psychological distress,adult attachment,and social support in primary caregivers of cancer patients.Methods:A total of 208 primary caregivers of cancer patients in one third grade hospital in Anhui Province were recruited.The 10-item Kessler Psychological Distress Scale(K10),Experiences in Close Relationships Inventory(ECR) and Social Support Questionnaire(SSQ) were used to explore psychological distress,adult attachment,and social support status.Results:The average K10 score was (21.5 ± 7.5).Multiple linear regression analysis indicated that caregiver gender,pressure on patient care,and attachment anxiety had positive prediction on psychological distress (β =2.30,3.02,0.13),while residence had negative prediction on psychological distress (β =-3.22).Conclusion:It suggests that the psychological distress is related to attachment anxiety and social support among the primary caregivers.

Adolescent ; Heart Ventricles ; injuries ; Humans ; Male ; Thoracic Injuries ; complications ; Ultrasonography ; Ventricular Septal Rupture ; diagnostic imaging ; etiology ; surgery ; Wounds, Nonpenetrating ; complications

Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode' s solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 micromol L(- 1)), but remained no change in the presence of Str (1 and 100 nmol L(-1)). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol L(-1)) , but had no obvious effects on the action of Str (10 micromol L(-1)). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 micromol L(-1)) or the removal of Ca2+ from Tyrode's solution. In Na+, K+ -free Tyrode' s solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 micromol L(-1)) was attenuated, while remained no change to Str (1 and 100 nmol L(-1)). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+ -free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+ -ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester ; pharmacology ; Aequorin ; pharmacology ; Animals ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Calcium Channels ; metabolism ; Fura-2 ; pharmacology ; supply & distribution ; Guinea Pigs ; Myocardium ; pathology ; Nifedipine ; pharmacology ; Ryanodine ; pharmacology ; Sarcolemma ; metabolism ; pathology ; Sarcoplasmic Reticulum ; drug effects ; metabolism ; Sodium-Calcium Exchanger ; Sodium-Potassium-Exchanging ATPase ; antagonists & inhibitors ; Strophanthidin ; pharmacology ; Tetrodotoxin ; pharmacology ; Thapsigargin ; pharmacology

OBJECTIVETo evaluate the quality of life (QOL) in patients with esophageal carcinoma after thoracoscopic and laparoscopic esophagectomy and circular stapled cervical esophagogastric anastomosis via retrosternal route or three-incision open surgery.

METHODSA total of 63 patients with middle-upper esophageal carcinoma who underwent radical surgical resection from January 2009 to October 2010 were enrolled in this study. Thirty-three patients underwent combined laparoscopic and thoracoscopic surgery and 30 three-incision open surgery. The EORTC questionnaire QLQ-C30 and QLQ-OES18 were used to evaluate the QOL.

RESULTSThere were no significant differences in the clinical data between the two groups except for anastomosis method(P>0.05). In the endoscopy group, there was one patient developed anastomotic leakage(3.0%, 1/33), 1 postoperative wound infection in the neck (3.0%, 1/33), and 1 anastomotic stricture(3.0%, 1/33). In the open group, 8 patients had anastomotic leakage (26.7%, 8/30), 2 had anastomotic stricture (6.7%, 2/30), 1 had wound infection in the neck (3.3%, 1/30), and 6 had pulmonary infection (20.0%, 6/30). All the complications were managed by conservative treatment. The two groups differed in dysphagia, food intake, pain, obstruction, dyspnea, anorexia, fatigue, financial condition, physical function, role function, emotional function, cognitive function, social function and global health level and were more favorable in the endoscopy group(P<0.05), while there were no significant differences in the other dimensions.

CONCLUSIONSThe postoperative complication rate is low after thoracoscopic and laparoscopic esophagectomy. Stapled anastomosis is associated with lower rate of anastomotic leak. QOL is better in patients following thoracoscopic and laparoscopic esophagectomy as compared to those following three-incision open surgery.

Aged ; Anastomosis, Surgical ; methods ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Esophagus ; surgery ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Middle Aged ; Quality of Life ; Stomach ; surgery ; Thoracoscopy

AIM:To investigate the role of prostaglandin E2receptor 2 agonist (EP2A) in proliferation and homing of human CD34 +cells. METHODS:Bone marrow fluid and peripheral blood containing stem cells were collected from healthy donors mobilized by granulocyte colony-stimulating factor in our department. Human CD34 +cells were isolated by the method of magnetic-activated cell sorting microbeads. Bone marrow mononuclear cells were isolated by Ficoll-Paque centrifugation,and the bone marrow mesenchymal stem cells(BMMSC) were cultured with L-DMEM. Human CD34 +cells and BMMSC were divided into 4 groups,and treated with PGE2(as positive control),DMSO(as negative control),EP2A and EP2A+prostaglandin E2receptor 2 antagonist (EP2AA),respectively. After exposed to the reagents,human CD34 +cell viability was measured by CCK-8 assay,the number of colonies was evaluated by colony-formation assay,the cell cycle distribution was analyzed by flow cytometry,and the protein expression of survivin,β-catenin and CXC chemokine receptor 4 (CXCR4) was detrmined by Western blot. Moreover, the concentration of stromal cell-derived factor-1α (SDF-1α) in the BMMSC was detected by ELISA. RESULTS:The cell viability and the colony number of human CD34 +cells in EP2A group were not higher than those in negative control group. Furthermore,the proportion of human CD34 +cells treated with EP2A in G2/M phase was not elevated compared with negative control group. The protein expression of survivin and β-cate-nin did not up-regulated in human CD34 +cells exposed to EP2A,but the protein expression of CXCR4 in human CD34 +cells and the concentration of SDF-1α in BMMSC were elevated. CONCLUSION:EP2A promotes human CD34 +cell homing in vitro but not proliferation.

To identify up-regulated genes in adult rat cardiac fibroblasts (CF) induced by angiotensin II (Ang II), suppression subtractive hybridization (SSH) was performed between the CF stimulated by Ang II (tester) and unstimulated CF (driver) to generate subtractive cDNA library. The library was screened with dot blots hybridization to further verify the differentially expressed cDNA clones. Partial positive clones (19 up-regulated genes) were sequenced and BLAST analyzed. Twelve up-regulated genes related to extracellular matrix, cell cycle, intracellular signal transduction, cell cytoskeleton, cell metabolism and 7 new expressed sequence tags (EST) were acquired (GenBank accession number: CN382808, CN382809, CN382810, CN382811, CN382812, CN382813, CN382814). Our data reveal that SSH is a powerful technique of high sensitivity for the detection and cloning of up-regulated genes expressed in CF induced by Ang II, which may be helpful to clarify the mechanism of cardiac remodeling.
Angiotensin II ; pharmacology ; Animals ; Cells, Cultured ; DNA, Complementary ; genetics ; Expressed Sequence Tags ; Fibroblasts ; cytology ; Gene Expression Regulation ; Male ; Myocardium ; cytology ; Rats ; Rats, Sprague-Dawley ; Up-Regulation ; Ventricular Remodeling ; genetics

OBJECTIVETo analyze the association between mutation(s) in preS region of HBV and hepatitis B disease progress in Chinese patients with genotype C chronic HBV infection.

METHODSNinety-three patients with chronic genotype C HBV infection, including 24 asymptomatic carriers (ASC), 26 patients with chronic hepatitis B (CHB), 22 patients with liver cirrhosis (LC) and 21 HCC patients were investigated. Levels of HBV DNA, HBeAg, alanine aminotransferase (ALT), asparate transaminase (AST) were measured. HBV preS region was analyzed by PCR direct sequencing.

RESULTSThe prevalence of preS T3098C and T53C mutations of genotype C HBV was significantly higher in LC and HCC patients than ASC and CHB patients. The rate of T3098C mutation in ASC, CHB, LC, and HCC patients were 0.00% (0/24), 3.85% (1/26), 9.09% (2/22), and 30.77% (8/22), respectively (P=0.0015), while the rate of T53C mutation was 12.50% (3/24), 3.85% (1/26), 40.91% (9/22), and 42.31% (11/26), respectively (P=0.0012).

CONCLUSIONThe frequency of genotype C HBV preS T3098C and T53C mutations is associated with hepatitis B infection progression

Adolescent ; Adult ; Aged ; DNA, Viral ; genetics ; Female ; Gene Expression Regulation, Viral ; physiology ; Genotype ; Hepatitis B ; pathology ; virology ; Hepatitis B virus ; classification ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Young Adult