HER2, a member of epidermal growth factor receptor family proteins, is overexpressed in about 30% of human breast cancer. Increased levels of HER2 are associated with poor patient prognosis and enhanced metastasis. RNA interference (RNAi) is developed recently as a new technique which can inhibit gene expression specifically in mammalian cells. On the basis of previous study,in which two target sequences with favorable RNAi effect on HER2 were identified, a series of dual promoter siRNA-expressing vectors containing two opposing U6 and H1 promoters were constructed. After transfection of HER2-overexpressing SKBR3 breast cancer cells with the siRNA-expressing vectors, downregulation of HER2 was identified quantitatively. Subsequently, the siRNA-expressing cassettes were subcloned into lentiviral vectors by LR recombination reaction and lentivirus was prepared successfully. The results from infection of SKBR3 cells with siRNA-expressing lentivirus demonstrated that lentiviral-mediated RNAi could downregulate HER2 expression efficiently through fluorescent quantitative PCR (FQ-PCR), western blot, and FACS analysis. Furthermore, cell growth was inhibited in cell proliferation assay after treatment with siRNA lentivirus.A new tool for clarifying the function of HER2 in cancer metastasis and developing the gene therapy drug was offered.

OBJECTIVETo compare the effect of transurethral resection of the prostate combined with endocrine therapy (TURP + ET) with that of αlA-blockers combined with ET ((αlA-b + ET) in the treatment of bladder outlet obstruction (BOO) in patients with advanced prostate cancer (PCa), and to investigate the safety of the TURP + ET for the treatment of PCa with BOO.

METHODSWe retrospectively analyzed 63 cases of PCa with BOO, 28 treated by αlA-b + ET and the other 35 by TURP + ET. We obtained the residual urine volume (RV), maximum urinary flow rate (Qmax), International Prostate Symptom Score (IPSS), and quality of life score (QoL) before and after treatment along with the overall survival rate of the patients, followed by comparison of the parameters between the two methods.

RESULTSAt 3 months after treatment, RV, IPSS, and QoL in the TURP + ET group were significantly decreased from (137.8 ± 27.6) ml, (22.3 ± 3.6), and (4.2 ± 0.8) to (29 ± 13.6) ml, (7.8 ± 2.1), and (1.6 ± 0.5) respectively (P < 0.05), while Qmax increased from (5.6 ± 2.1) ml/s to (17.6 ± 2.7) ml/s (P < 0.05); the former three parameters in the αlA-b + ET group decreased from (133.6 ± 24.9) ml, (21.5 ± 3.2), and (4.7 ± 1.1) to (42 ± 18.3) ml, (12.8 ± 2.6), and (2.5 ± 0.7) respectively (P < 0.05), while the latter one increased from (6.3 ± 2.4) ml/s to (11.7 ± 2.3) ml/s (P < 0.05), all with statistically significant differences between the two groups (P < 0.05). The overall survival rate of the TURP + ET group was not significantly different from that of the αlA-b + ET group (51.4% vs 46.4% , P > 0.05).

CONCLUSIONTURP + ET is preferable to αlA-b + ET for its advantage of relieving BOO symptoms in advanced PCa without affecting the overall survival rate of the patients.

Adrenergic alpha-1 Receptor Antagonists ; therapeutic use ; Antineoplastic Agents, Hormonal ; therapeutic use ; Combined Modality Therapy ; methods ; Humans ; Male ; Prostatic Neoplasms ; complications ; drug therapy ; pathology ; surgery ; Quality of Life ; Retrospective Studies ; Transurethral Resection of Prostate ; Treatment Outcome ; Urinary Bladder Neck Obstruction ; drug therapy ; etiology ; surgery

OBJECTIVETo investigate the protective effect of lycopene against cryopreservation injury of post-thawing human sperm and its mechanism.

METHODSSemen samples were collected from 25 volunteers, each sample equally divided into four parts to be cryopreserved with cryoprotectant only (Ly0 control) or cryoprotectant + lycopene at the concentrations of 2 (Ly2), 5 (Ly5), and 10 µmol/L (Ly10), respectively. Before and after thawing, the semen samples were subjected to computer-assisted semen analysis ( CASA) for sperm kinematics, flow cytometry for sperm apoptosis, thiobarbituric acid assay for malondialdehyde (MDA) concentration, and JC-1 fluorescent staining for the sperm mitochondrial membrane potential (MMP).

RESULTSAfter cryopreservation, sperm motility was markedly decreased in all the groups (P < 0.01). The rate of sperm apoptosis was significantly lower in the Ly5 group than in the Ly0 control ([25.68 ± 4.36]% vs [33.26 ± 4.78]%, P < 0.05), while sperm MMP remarkably higher in the former than in the latter ([66.18 ± 14.23]% vs [55.24 ± 12.31]%, P < 0.05). The Ly2, Ly5 and Ly10 groups showed no statistically significance differences in the MDA level from the Ly0 control (P > 0.05).

CONCLUSIONAddition of lycopene at a proper concentration to cryoprotectant may reduce oxidative damage to sperm mitochondria in the freezing-thawing process, attenuate oxidative stress injury induced by reactive oxygen species to sperm plasma membrane, and improve the anti-apoptosis ability of sperm.

Apoptosis ; Carotenoids ; pharmacology ; Cryopreservation ; Cryoprotective Agents ; pharmacology ; Flow Cytometry ; Humans ; Male ; Malondialdehyde ; analysis ; Oxidative Stress ; Reactive Oxygen Species ; Semen Analysis ; Semen Preservation ; adverse effects ; methods ; Sperm Motility ; Spermatozoa ; drug effects ; physiology

Objective To explore the impact of chronic complications of type 2 diabetic mellitus (T2DM)on quality of life(QOL)among T2DM outpatients in urban China.Methods A cross.Sectional study was carried Out in 1524 T2DM outpatients recruited from 15 hospitals in 4 major cities of China.Questionnaire interviews were used to collect data on general characters and complications of T2DM.SF-36 questionnaire(version 1)was used and self-completed by patients under the help from interviewers for those having difficulties in reading.T test,one-way ANOVA and generalized linear model were used to explore the association between complications of T2DM and quality of life.Results Complication Was a significant predictor leading to poorer SF-36 subscale scores(decreased by 4.68-16.06 scores)and two summary scores (PCS,MCS).The scores of role-physical subscale(decreased by 22.45％)and role-emotionsl subscale(decreased by 16.28％)decreased much more than the other subscales in patients with complication.Overall,QOI Was reduced by 11.02％in patients with complication.The QoL had a gradual decrease with the increasing number of complications.The average scores descended from 2.82 to 10.33 in the eight subscales with the increase of one complication.Conclusion Having complication or multiple complications seemed to be predietors of poor QOL among T2DM outpatients.

OBJECTIVETo identify the risk factors of benign cervical anastomotic strictures after esophagectomy.

METHODSClinical data of 946 esophageal cancer patients undergoing esophagectomy with cervical anastomosis between 2003 and 2012 were analyzed retrospectively. Benign stricture was defined as dysphagia for which endoscopic dilation of the anastomosis was needed. Histologically proven malignant stricture was not regarded as benign stricture. χ(2) test and logistic regression model were used for univariate and multivariate analysis respectively.

RESULTSA total of 146 patients(16.5%) developed benign stricture during follow-up. Univariate analysis showed that the patients with cardiovascular disease (P=0.001), diabetes mellitus(P=0.041), gastric tube reconstruction(P=0.050), end-to-end anastomosis (P=0.013), or postoperative anastomotic leakage(P=0.008) had higher stricture rate. Multivariate analysis revealed that cardiovascular disease(P=0.004), gastric tube reconstruction (P=0.026), end-to-end anastomosis(P=0.043), and postoperative anastomotic leakage(P=0.001) were independently predictive factors for development of benign stricture.

CONCLUSIONSThe benign cervical stricture rate after esophagetomy with cervical gastric anastomosis is quite high. In order to prevent benign stricture formation, end-to-end anastomosis should be avoid. Blood pressure should be controlled for those with cardiovascular disease. Endoscopic dilation in an earlier stage postoperatively should be considered for those who develop anastomotic leakage.

Anastomosis, Surgical ; adverse effects ; Anastomotic Leak ; Constriction, Pathologic ; etiology ; Deglutition Disorders ; etiology ; Esophageal Neoplasms ; complications ; surgery ; Esophageal Stenosis ; etiology ; Esophagectomy ; adverse effects ; Follow-Up Studies ; Humans ; Postoperative Complications ; etiology ; Reconstructive Surgical Procedures ; adverse effects ; Retrospective Studies ; Risk Factors

To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established.
Naphthaleneacetic Acids ; pharmacology ; Phenylurea Compounds ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Shoots ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Thiadiazoles ; pharmacology ; Tissue Culture Techniques ; Tulipa ; drug effects ; growth & development

OBJECTIVETo observe the influence of endoscopic radial artery harvesting techniques on the prevalence of complications after coronary artery bypass grafting, and to assess the potential trauma to the radial artery through the histological changes.

METHODSFrom August 2003 to June 2008, 87 patients undergoing CABG had radial artery harvested by endoscopic harvesting system. About 4 mm proximal and distal radial artery end segment of 10 patients undergoing endoscopic and conventional harvesting were examined with light and electro-microscope.

RESULTSThe endoscopic harvest time was 42 to 98 min, with a mean of (57.6 +/- 17.3) min. The harvested conduit length was 15 to 20 cm, with a mean of (17.5 +/- 1.6) cm. Objective dorsal thenar numbness remained in 7 patients, none complained of forearm numbness at 3-month follow-up. The result of light and electro-microscope had no differences in the intima, media, or adventitia between endoscopically and conventionally obtained radial artery segments.

CONCLUSIONThe use of endoscopic radial artery harvesting in coronary artery bypass grafting can be performed safely with infrequent complications. This method results in excellent patient satisfaction, particularly regarding the cosmetic outcome.

Adult ; Aged ; Coronary Artery Bypass ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; transplantation ; Tissue and Organ Harvesting ; methods ; Treatment Outcome

OBJECTIVETo assess the efficacy of endoscopic epicardial microwave ablation, a new completely endoscopic technique, for isolated atrial fibrillation (AF).

METHODSFrom July to October of 2004, 5 patients with isolated permanent AF underwent this procedure. We performed epicardial microwave ablation using FLEX10 probe under the guide of VasoView 5 endoscope. The probe was placed around left and right pulmonary vein orifices, the microwave application was set 65 W/90 s, and a continuous ablation line was obtained encircling the pulmonary veins. Echocardiography and electrocardiography were carried out in all patients before and after operation.

RESULTSFive patients received electrical conversion after ablation. Average ablation time was 35.0 min and average procedure time was 2.5 h without any procedure-related complications. At the 3-month, 6-month and 12-month follow-up, 4 patients (80%) were in sinus rhythm, associated with a reduced left atrial diameter and an improved cardiac function.

CONCLUSIONEndoscopic epicardial microwave ablation for isolated atrial fibrillation appears to be safe and effective.

Adult ; Atrial Fibrillation ; surgery ; Catheter Ablation ; methods ; Endoscopy ; Female ; Follow-Up Studies ; Humans ; Male ; Microwaves ; therapeutic use ; Middle Aged ; Treatment Outcome

OBJECTIVETo apply the off-pump epicardial microwave ablation (MW) to treat the permanent atrial fibrillation (PAF) and evaluate the clinical effect.

METHODSFrom April 2003 to April 2005, a total of 46 patients with PAF underwent off-pump epicardial MW, including the patients with mitral valve disease (28 cases), mitral and aortic valve disease (5 cases), coronary artery disease (8 cases) and lone PAF (5 cases). FLEX 10 probe was placed around four pulmonary orifices and the access orifice of the inferior vena cava to tricuspid annulus. The microwave application was set 65 W, 90 s and a continuous ablation line was finished encircling the pulmonary veins. Subsequently, patients underwent either off-pump coronary artery bypass graft or valve surgery on-pump. Mean left atrial diameter was (52.5 +/- 15.3) mm, LVEF 42% - 70% before operation. All patients were followed up in the discharge, 3, 6, 12 months after surgery by standard 12-lead ECG, 24 h Holter, UCG and clinical examination.

RESULTSAll procedures were completed successful on the off-pump cardiac surgery and 46 patients were converted sinus rhythm when the operations were performed. Mean ablation time was (37.7 +/- 7.8) min, without any related complications. At discharge, 3, 6 and 12 months follow-up, 63.0%, 66.7%, 72.7% and 80.6% of the patients kept sinus rhythm and left atrial diameter of < 50 mm.

CONCLUSIONSUsing MW, electrical isolation of the pulmonary veins can be achieved epicardially without cardiopulmonary bypass support. Off-pump epicardial microwave ablation is an efficient option with an excellent benefit/risk ratio for the treatment of PAF with or without other cardio-surgical disease.

Adult ; Aged ; Atrial Fibrillation ; surgery ; Catheter Ablation ; methods ; Coronary Artery Bypass, Off-Pump ; Female ; Follow-Up Studies ; Humans ; Male ; Microwaves ; therapeutic use ; Middle Aged ; Pulmonary Veins ; surgery ; Treatment Outcome

OBJECTIVETo evaluate four candidate variable number tandem repeat (VNTR) loci for genotyping Mycobacterium tuberculosis complex strains.

METHODSGenomic sequences for two M. tuberculosis strains (CCDC5079 and CCDC5180) were generated, and using published sequence data, four candidate VNTR loci were identified. The VNTRs were used to genotype 225 Chinese clinical M. tuberculosis complex strains. The discriminatory power of the VNTRs was evaluated using BioNumerics 5.0 software.

RESULTSThe Hunter-Gaston Index (HGI) for BJ1, BJ2, BJ3, and BJ4 loci was 0.634, 0.917, 0.697, and 0.910, respectively. Combining all four loci gave an HGI value of 0.995, thus confirming that the genotyping had good discriminatory power. The HGI values for BJ1, BJ2, BJ3, and BJ4, obtained from Beijing family strain genotyping, were 0.447, 0.878, 0.315, and 0.850, respectively. Combining all four loci produced an HGI value of 0.988 for genotyping the Beijing family strains. We observed unique patterns for M. bovis and M. africanum strains from the four loci.

CONCLUSIONWe have shown that the four VNTR loci can be successfully used for genotyping M. tuberculosis complex strains. Notably, these new loci may provide additional information about Chinese M. tuberculosis isolates than that currently afforded by established VNTR loci typing.

Cluster Analysis ; Genotyping Techniques ; Humans ; Minisatellite Repeats ; Mycobacterium bovis ; genetics ; Mycobacterium tuberculosis ; genetics