OBJECTIVETo investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer.

METHODSRecombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined.

RESULTSThe viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group.

CONCLUSIONThe combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; genetics ; Prostatic Neoplasms ; blood supply ; pathology ; Transfection

OBJECTIVESTo analyze the mood factors in prostatitis and evaluate the effects of Molida psychological therapy.

METHODSTwo hundred and thirty-six chronic prostatitis patients were divided according to the course of disease into above 6 months group (n = 31) and below 6 months group(n = 205) as well as into sexual disease group(n = 25) and non-sexual disease group(n = 211). An investigation was made by self-rating method on the basis of the National Institutes of Health Chronic Prostatitis Symptom Index(NIH-CPSI) and Symptom Checklist 90 (SCL-90) and 56 cases were rated again after Molida therapy.

RESULTS1. The scores of SCL-90 in 236 prostatitis patients were significantly higher than normal(P < 0.01). The factor scores of SCL-90 showed one-item positive in 107 patients (45.2%), of whom 27 (25.2%) had depressive disorder and 80(74.77%) had anxiety (23 with significant compulsion). Thirty-eight cases(16.1%) were two items positive. 2. The scores of SCL-90 were significantly higher in the > 6 months group of history and the sexual disease group than in the control group (P < 0.01). 3. The scores of NIH-CPSI showed a positive correlation with those of SCL-90 and both scores in 43/56 cases were significantly decreased after psychological treatment (P < 0.01).

CONCLUSIONSThe mood factor plays an important role in aggravating symptoms in chronic prostatis patients and causes difficulty for management. Molida may significantly improve the mood and symptoms of the chronic prostatitis patient.

Adult ; Affect ; Chronic Disease ; Humans ; Male ; Middle Aged ; Prostatitis ; psychology ; therapy ; Psychotherapy

OBJECTIVETo investigate the exposure levels of carbon disulfide (CS(2)) for a chemical fiber industry.

METHODSThe concentration of CS(2) was monitored in representative workshops and types of work, and the datas of that over the years were collected.

RESULTSThe short-term exposure concentration of CS(2) about 80% of the type of work was less than or equal to 10 mg/m(3), which of more than 90% was less than or equal to 20 mg/m(3). The time weighted average concentration of CS(2) about 70% of the type of work was less than or equal to 5 mg/m(3), which of more than 90% was less than or equal to 10 mg/m(3). The short-term exposure concentration of CS(2) which was more than 15 mg/m(3) or the time weighted average concentration of CS(2) which was more than 30 mg/m(3) was only for little type of work.

CONCLUSIONThe concentration of CS(2) for the most type of work was lower, but there were still a number of types of work exposuring the higher concentration, which exceed the national occupational exposure limits.

Carbon Disulfide ; analysis ; Chemical Industry ; Humans ; Occupational Exposure ; analysis ; Workplace

OBJECTIVETo study the effects of long-term exposure to carbon disulfide (SC(2)) on cardiovascular system of workers.

METHODSThe concentrations of CS(2) were detected in the representative workshops with different exposure levels. The indicators related to cardiovascular system were tested in 633 workers occupationally exposed to CS(2), which included blood pressure, electrocardiogram, blood routine (blood RT), cholesterol (TCHO), triglyceride (TG) and so on. The data were analyzed by chi-square test and multiple logistic regression analysis.

RESULTSThe exposure concentration of CS(2) for 389 workers was less than or equal to 5 mg/m(3), which for other 244 workers was higher than 5 mg/m(3). The maximum exposure concentration of CS(2) was 15.73 mg/m(3). There were no significant effects of CS(2) on the electrocardiogram, red blood cells, white blood cells, blood platelet, TCHO and TG of workers. However, the positive effects of CS(2) on blood pressure and negative effects of CS(2) on hemoglobin were found. The rates of high TCHO, TG and hypertension in male workers were significantly higher than those in female workers (P < 0.05). The rates of high TCHO, hypertension and sinus arrhythmia in older workers (≤ 30 years old) were significantly higher than those in young workers (> 30 years old) (P < 0.05). The rate of sinus arrhythmia in workers with 1 - 10 working years was significantly higher than that in workers with more than 10 working years (P < 0.05). The rate of hypertension in workers with 1 - 10 working years was significantly lower than that in workers with more than 10 working years (P < 0.05).

CONCLUSIONThere were no significant effects of CS(2) exposure on the indexes of cardiovascular system of workers.

Adult ; Blood Pressure ; Carbon Disulfide ; adverse effects ; Cardiovascular System ; drug effects ; physiopathology ; Erythrocyte Count ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; analysis ; Risk Factors ; Young Adult

OBJECTIVETo understand the molecular basis for a potential reaction mechanism and develop novel antibiotics with homology modeling for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase (HMGS).

METHODSThe genetic engineering technology and the composer module of SYBYL7.0 program were used, while the HMGS three-dimensional structure was analyzed by homology modeling.

RESULTSThe mvaS gene was cloned from Streptococcus pneumoniae and overexpressed in Escherichia coli from a pET28 vector. The expressed enzyme (about 46 kDa) was purified by affinity chromatography with a specific activity of 3.24 micromol/min/mg. Optimal conditions were pH 9.75 and 10 mmol/L MgCl2 at 37 degrees C. The V(max) and K(m) were 4.69 micromol/min/mg and 213 micromol/L respectively. The 3D model of S. pneumoniae HMGS was established based on structure template of HMGS of Enterococcus faecalis.

CONCLUSIONThe structure of HMGS will facilitate the structure-based design of alternative drugs to cholesterol-lowering therapies or to novel antibiotics to the Gram-positive cocci, whereas the recombinant HMGS will prove useful for drug development against a different enzyme in the mevalonate pathway.

Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Gene Expression Regulation, Bacterial ; physiology ; Hydroxymethylglutaryl-CoA Synthase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Mutation ; Protein Conformation ; Streptococcus pneumoniae ; enzymology ; genetics

OBJECTIVETo investigate the management of complicated, severe or recurrent Budd-Chiari syndrome.

METHODSFrom February 2004 to August 2007, 28 patients with complicated, severe or recurrent Budd-Chiari syndrome were treated. In this series, 16 patients relapsed after treated with percutaneous transluminal angioplasty or stent deployment, 2 cases relapsed after surgery; and the other 10 were under severe conditions and hard to treat, including malignancy of the inferior vena cava and right atrium. Meso-cavo-atrial shunt was carried out in 10 cases, meso-cavo-jugular shunt in 6 (capitis medusa was used in one case), cavoatrial shunt in 2 and cavo-jugular shunt in 1, mesocaval shunt in 2, and radical or extended radical correction in 7.

RESULTSOne patient (3.6%) died in 24 hours after operation. Graft infection occurred in 1 case. Excellent, good, fair, poor and death rate were 22.2%, 55.5%, 14.8%, 3.7% and 3.7%, respectively, the overall effective rate was 92.5%.

CONCLUSIONTo select personalized treatment according to the disease status brings hopes to difficult, severe, recurrent Budd-Chiari syndrome.

Adolescent ; Adult ; Blood Vessel Prosthesis Implantation ; Budd-Chiari Syndrome ; surgery ; Child ; Child, Preschool ; Critical Illness ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Portacaval Shunt, Surgical ; Recurrence ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo explore a method for the treatment of the skin defects at the distal phalanges of 2-5th fingers.

METHODSThe island flap at the dorsum of the middle phalange was designed with the pedicle of dorsal branches from the digital proper artery. When the flap was used to repair defect at finger pulp, the dorsal branch of the digital proper nerve in the flap was kept to be anastomosed to the digital proper nerve at the recipient finger. From Feb. 2005 to May. 2010, 54 cases with skin defects at the distal phalanges of 61 fingers were treated with the flap, including 35 defects at finger pulp and 26 defects at finger tip.

RESULTSThe maximum size of defects and flaps was 2.2 cm x 2.5 cm and 2.4 cm x 2.7 cm, respectively. 61 flaps survived completely. Blister was happened in 3 flaps 2 days after operation, which healed spontaneously without necrosis. 54 cases were followed up for 5 to 22 months (average, 11 months). The flaps had good texture and color match with normal sensation (grade S4). The 2-point discrimination distance was 6-9 mm. The interphalangeal joint had normal movement.

CONCLUSIONSThe island flap at the dorsum of the middle phalange is an ideal method for the skin defect at the distal phalange of finger.

Adolescent ; Adult ; Aged ; Arteries ; Female ; Fingers ; blood supply ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps ; blood supply ; Young Adult

OBJECTIVETo elucidate the expression and function of VAP-33 gene in dendritic cell sarcoma (DCS) cell line.

METHODSThe expression of VAP-33 in DCS cells was investigated by mass spectrum with immunoprecipitation membrane protein. DCS cells were treated with antigens in different dosages (150, 850, and 1500 microl) for 24, 48 and 72 h respectively. Cell morphology and phagocytosis activity of DCS cells were measured. Indirect immunofluorescence, confocal microscopy and Western blotting were used to study the distribution and expression changes of VAP-33. Moreover, DCS cells were treated with 0.5 mol/L insulin for 20 min first and followed by Western blotting to detect changes of VAP-33 and glucose transfer protein 4 (GLUT-4) in the total cellular protein, cytoplasmic protein and membrane protein. Confocal microscopy was used to document the expression and distribution changes of VAP-33 and GLUT-4 in DCS cells.

RESULTSVAP-33 expression was obtained at the cell membrane and in the cytoplasm of DCS cells. Upon antigen stimulation, DCS cells showed more active phagocytosis and morphologically became more elongated with branched protrusions. The expression of VAP-33 was decreased by the antigen stimulation. Upon the insulin stimulation, the expression of VAP-33 and GLUT-4 were increased and co-localized.

CONCLUSIONSVAP-33 expression in DCS originated from the dendritic cells (DC) seemed relating to the vesicle transportation during antigen processing in DC. Additionally, VAP-33 and GLUT-4 also take part in the glucose transportation in the cells.

Animals ; Antigen Presentation ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cytoplasm ; metabolism ; Dendritic Cell Sarcoma, Interdigitating ; metabolism ; pathology ; Down-Regulation ; Glucose Transporter Type 4 ; metabolism ; Insulin ; pharmacology ; Membrane Proteins ; metabolism ; Mice ; Phagocytosis ; immunology

OBJECTIVETo observe the effects of antisense RNA of connective tissue growth factor (CTGF) on rat liver fibrosis.

METHODSGene recombinant techniques were used to construct a rat antisense RNA of CTGF recombinant plasmid which could be expressed in eukaryotic cells. The recombinant plasmids were encapsulated with lipofectamine and then transducted into a carbon tetrachloride (CCl4) induced rat liver fibrosis model. Expression of CTGF was assessed by RT-PCR, Western blot and immunohistochemistry. Immunohistochemistry was used to identify type I and III collagens. HE stained liver slides were used for pathological study.

RESULTSThe mRNA and protein expression of CTGF in the fibrotic liver transfected with antisense-CTGF were significantly decreased compared with those of the controls (P<0.01). The depositions of type I and type III collagens were also decreased (P<0.05). Antisense-CTGF also minimized the pathological fibrosis in the rat livers (P<0.01).

CONCLUSIONThe results demonstrate that the antisense RNA of CTGF recombinant plasmid has certain effects in preventing liver fibrosis and makes it a possible candidate for use in future gene therapy.

Animals ; Connective Tissue Growth Factor ; genetics ; Genetic Therapy ; Liver ; pathology ; Liver Cirrhosis, Experimental ; pathology ; Male ; Plasmids ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo investigate the role that E-cadherin (E-cad) plays on cell adhesion and proliferation of human breast carcinoma.

METHODSE-cad expression vector was transfected into an E-cad-negative human breast carcinoma MDA-MB-231 cells. G418 was used to screen positive clones. E-cad, β-catenin (β-cat) and cyclin D1 expressions of these clones were confirmed by Western blot. Their cell-cell and cell-matrix adhesion abilities were detected. E-cad/β-catenin interaction was confirmed by immunoprecipitation. Cell proliferation was evaluated by MTT. Cell apoptosis was analyzed by flow cytometry. Direct two-step immunocytochemistry was used to detect the localization of β-cat.

RESULTE-cad(+) cell strains Ecad-231-7 and Ecad-231-9 were established. When cultured in ultra-low-binding dishes Ecad-231 cells grow in suspension while Ecad-231-7 and Ecad-231-9 cells grow in large clamps. When co-cultured with HCT116 cells, the average adhesion rates at 30 min are 39.0%, 60.0% and 59.5% for MDA-MB-231, Ecad-231-7 and Ecad-231-9 respectively. The average detachment rates by EDTA for 5 min are 37.4%, 4.2% and 7.4% respectively. So E-cad expression enhanced hemotypic and heterotypic cell-cell adhesion and cell-matrix adhesion. Forced exogenously expressed E-cad could combine with endogenous β-cat, whereas down stream cyclin D1 expression was significantly decreased, as evidenced by Western blot. The rates of cell apoptosis of MDA-MB-231, Ecad-231-7 and Ecad-231-9 were 1.8%, 2.0% and 2.1%. Expression of E-cad had no obvious effect on the apoptosis of tumor cells with regular culture. β-cat increased in the cytoplasma.

CONCLUSIONSTwo monoclonal tumor cell strains (Ecad-231-7 and Ecad-231-9) stably expressing E-cad were successfully established. E-cad could enhance adhesion and inhibit proliferation of human breast carcinoma cells through a pathway involving β-cat and cyclin D1.

Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cadherins ; genetics ; metabolism ; physiology ; Cell Adhesion ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Female ; Genetic Vectors ; Humans ; Plasmids ; Transfection ; beta Catenin ; metabolism