The aim of this paper is to report the study on gene silencing efficiency of siRNA targeted against mouse VEGFR2 (siVEGFR2) in vitro mediated by polyethyleneimine (PEI) and its anti-tumor effect in vivo. CY3-labeled siRNA was compounded into PEI and transfected into MS1 cells. Confocal microscopy was used to image the subcellular distribution of siRNA in MS1 cells. Semi-quantitative RT-PCR and Western blotting were used to evaluate VEGFR2 gene silencing induced by siVEGFR2/PEI complexes. A tumor-bearing nude mice model was established to compare the anti-tumor effect after delivered by local and systemic routes. siVEGFR2/PEI complex-transfected cells exhibited much fluorescence in cytoplasm with no evidence of nuclear accumulation. The expression levels of VEGFR2 mRNA and protein in PEI-transfected cells were significantly down-regulated compared with that in blank group, the silencing efficiency were 28.2% and 23.6% respectively. The tumor sizes in mice intratumorally injected with siVEGFR2/PEI complexes (189.429 +/- 17.562 mm3) were reduced definitely compared to that in mice injected with siVEGFR2/PEI complexes via vein route (315.507 +/- 20.491 mm3), or to saline groups (365.844 +/- 20.713 mm3). The study demonstrated that PEI could effectively transfect siRNA into cells and silence the VEGFR2 gene expression. Intratumoral delivery is more suitable for non-targeted modified PEI/siRNA complexes to inhibit the tumor growth in vivo. The present data lay a solid foundation to further study on the gene silencing mechanism for PEI-medicated RNAi and its anti-tumor efficiency in vivo.

BACKGROUND: The recent development of stem cells has provided new ideas for the treatment of androgen-deficient diseases. OBJECTIVE: To investigate whether adipose-derived mesenchymal stem cells can differentiate into Leydig cells.METHODS: Passage 3 rat adipose-derived mesenchymal stem cells that grew well were taken and cultured in the medium with (experimental) or without (control) 0.1 mg/L human chorionic gonadotropin, 10.0 μg/L platelet-derived growth factor and 10.0 μg/L basic fibroblast growth factor. Indicator detection was done at 1, 7, 14, 24 days of induced culture. RESULTS AND CONCLUSION: (1) Immunofluorescence staining results showed that there were no 3β-hydroxysteroid dehydrogenase (3β-HSD) positive cells in the control group, while the number of 3β-HSD positive cells was gradually increased in the experimental group with the induction time, which presented with fluorescence enhancement. (2) There was no secretion of testosterone in the control group, while in the experimental group, testosterone secretion was detected at 7 days of induced culture, and moreover, the testosterone level was increased with the induction time. (3) RT-PCR findings showed no luteinizing hormone receptor, steroidogenic acute regulatory protein, and 3β-HSD positive bands in the control group, while these positive bands appeared in the experimental group after 1 day of induction, and strengthened with the induction time. To conclude, adipose-derived mesenchymal stem cells can be induced to differentiate into Leydig cells.

OBJECTIVETo construct an eukaryotic expression vector of open reading frame of unknown KH gene (KH-ORF), and investigate its effect on cell proliferation.

METHODSThe pCI-neo-KH-ORF expression vector was constructed by DNA recombinant technique and was introduced into COS-7 cells and K562 cells by lipofectactin-mediated DNA transfection. Expression of KH-ORF mRNA was detected by RT-PCR. The effect of KH-ORF on cell cycle of COS-7 cells and K562 cells was evaluated by flow cytometry (FCM). Effect on cell proliferation of COS-7 cells was tested by MTT assay and that on K562 cells was analyzed by growth curves and LDH activity measurement.

RESULTS(1) KH-ORF mRNA was expressed both in COS-7 cells and K562 cells. (2) The cell cycle and cell proliferation of COS-7 cells were unaffected significantly. (3) The proportion of cells in S phase was increased in pCI-neo-KH-ORF-transfected K562 cells; and growth curves and LDH activity indicated enhanced cell proliferation.

CONCLUSIONKH gene may be a leukemia gene related to proliferation of K562 cells.

Animals ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; Gene Expression Regulation, Neoplastic ; Genes, Neoplasm ; genetics ; physiology ; Genetic Vectors ; Humans ; K562 Cells ; L-Lactate Dehydrogenase ; metabolism ; Open Reading Frames ; genetics ; physiology ; Plasmids ; RNA, Messenger ; biosynthesis ; genetics ; RNA-Binding Proteins ; genetics ; S Phase ; Transfection

The aim of this paper is to report the study on gene silencing efficiency of siRNA targeted against mouse VEGFR2 (siVEGFR2) in vitro mediated by polyethyleneimine (PEI) and its anti-tumor effect in vivo. CY3-labeled siRNA was compounded into PEI and transfected into MS1 cells. Confocal microscopy was used to image the subcellular distribution of siRNA in MS1 cells. Semi-quantitative RT-PCR and Western blotting were used to evaluate VEGFR2 gene silencing induced by siVEGFR2/PEI complexes. A tumor-bearing nude mice model was established to compare the anti-tumor effect after delivered by local and systemic routes. siVEGFR2/PEI complex-transfected cells exhibited much fluorescence in cytoplasm with no evidence of nuclear accumulation. The expression levels of VEGFR2 mRNA and protein in PEI-transfected cells were significantly down-regulated compared with that in blank group, the silencing efficiency were 28.2% and 23.6% respectively. The tumor sizes in mice intratumorally injected with siVEGFR2/PEI complexes (189.429 +/- 17.562 mm3) were reduced definitely compared to that in mice injected with siVEGFR2/PEI complexes via vein route (315.507 +/- 20.491 mm3), or to saline groups (365.844 +/- 20.713 mm3). The study demonstrated that PEI could effectively transfect siRNA into cells and silence the VEGFR2 gene expression. Intratumoral delivery is more suitable for non-targeted modified PEI/siRNA complexes to inhibit the tumor growth in vivo. The present data lay a solid foundation to further study on the gene silencing mechanism for PEI-medicated RNAi and its anti-tumor efficiency in vivo.
Animals ; Cell Line, Tumor ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Female ; Gene Silencing ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Polyethyleneimine ; chemistry ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; metabolism ; Spleen ; cytology ; Transfection ; Tumor Burden ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

Adult ; Aged ; Female ; Hepatitis B, Chronic ; complications ; Hepatorenal Syndrome ; diagnosis ; etiology ; therapy ; Humans ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo explore the effect of prostatectomy on nocturia in patients with benign prostatic hyperplasia (BPH).

METHODSThe data of patients who had received prostatectomy for BPH between June 2006 and December 2007 were collected. Nocturia severity was assessed preoperatively and 3 to 6 months after prostatectomy by the number of nocturia events, the time from falling sleep to the first awakening to void (hours of undisturbed sleep, HUS), the score of the nocturia quality of life (N-QOL) questionnaire, the International Prostatic Symptom Score (IPSS) and the quality of life (QOL) score.

RESULTSOne hundred and twenty five cases were included. Of them, 73 patients finished the follow-up completely. There were 62 patients whose number of nocturia events before the operation was equal or more than 2. The data from these 62 patients were analyzed. Of them, 56 patients underwent transurethral resection of prostate, the remaining 11 patients suprapubic prostatectomy. Significant improvement (P < 0.01) was noted in all the following parameters after treatment: the number of nocturia events decreased from 4.2 ± 2.4 to 2.2 ± 1.0, HUS increased from (1.8 ± 0.7) h to (3.0 ± 1.4) h, N-QOL score raised from 30 ± 10 to 40 ± 7, IPSS decreased from 23 ± 5 to 8 ± 5, and QOL score fell down from 4.4 ± 0.7 to 1.5 ± 1.0.

CONCLUSIONThe prostatectomy can markedly improve the symptoms of nocturia, sleep and life quality in the BPH patients who accompanied with nocturia.

Aged ; Aged, 80 and over ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Nocturia ; complications ; surgery ; Prostatectomy ; Prostatic Hyperplasia ; complications ; surgery ; Quality of Life ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo Explore the effect of oblique preputial island flap for the treating of hypospadias.

METHODSFifty-one patients were performed one-stage urethroplasty with oblique preputial island flap to repair hypospadias.

RESULTSAll cases resulted in a good contour of the penis without any redundancy and a normal anatomic position of slit-shaped urethral meatus. The urination was perfect. Six patients occurred complications (3 cases of urinary fistula, 3 cases of meatal stenosis).

CONCLUSIONWith extensive scope of materials, reliable blood supply of skin flap, satisfactory appearance of shaping penis and few complications, one-stage urethroplasty with oblique preputial island flap is an effective method to repair hypospadias of penile type and penile-scrotal type.

Adolescent ; Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Male ; Penis ; surgery ; Surgical Flaps ; Urethra ; surgery

BACKGROUNDSurgical treatment options for patients with cirrhosis and portal hypertension are complicated. In this study, we evaluated the effectiveness of a new treatment strategy, splenic auto-transplantation and oesophageal transection anastomosis. We report results from clinical observations, splenic immune function and portal dynamics in 274 patients.

METHODSFrom 1979 to 2005, 274 cirrhosis patients with portal hypertension underwent the new treatment strategy, and were followed up to compare results with those patients who underwent traditional surgical treatment. From 1999 to 2002, a randomized controlled trial (RCT) was performed on 40 patients to compare their post-operative immune function. From 1994 to 2006, another RCT enrolled 28 patients to compare portal dynamics using three-dimensional dynamic contrast-enhanced magnetic resonance angiography (3D DEC MRA) investigation post operation.

RESULTSAmong 274 patients (mean age 41.8 years), the emergency operative mortality (4.4%), selective operative mortality (2.2%), complication rate (17.9%), prevalence of hepatic encephalopathy (< 1%), rate of portal hypertension gastritis (PHG) bleeding (9.1%), and morbidity of hepatic carcinoma (8%) were similar to those patients undergoing traditional operation; the spleen immunology function (Tuftsin, IgM) decreased in both groups 2 months post operation, but this decrease did not reach statistical significance. Through 3D DCE MRA, the cross sectional area and the velocity and volume of blood flow of the main portal vein decreased significantly after operation in both groups. The velocity and volume of blood flow in the auto-transplantation group was significantly lower than that in the control group.

CONCLUSIONSSplenic auto-transplantation and esophageal transection anastomosis is a safe, effective, and reasonable treatment strategy for patients with portal hypertension with varicial bleeding. It not only can correct hypersplenism, but may also achieve complete hemostasis. Spleens auto-transplanted into the retroperitoneal space can preserve immune function and establish broad collateral circulation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anastomosis, Surgical ; Child ; Esophagus ; surgery ; Female ; Humans ; Hypertension, Portal ; immunology ; surgery ; Imaging, Three-Dimensional ; Immunoglobulin M ; blood ; Male ; Middle Aged ; Prospective Studies ; Spleen ; transplantation ; Transplantation, Autologous

BACKGROUNDCD4 count is used to determine antiretroviral therapy (ART) eligibility. In China, flow cytometers are mostly located in urban areas with limited access by patients residing in remote areas. In an attempt to address this issue, we conducted a study to validate the performance of Alere PIMA point-of-care CD4 analyzer.

METHODSVenous and finger-prick blood specimens were collected from HIV-positive participants from two voluntary counseling and testing sites in Yunnan Province. Both venous and finger-prick blood specimens were tested with the PIMA analyzer. Venous blood specimens tested with the Becton Dickinson FACSCalibur were used as a reference.

RESULTSVenous specimens from 396 and finger-prick specimens from 387 persons were available for analysis. CD4 counts by PIMA correlated well with those from FACSCalibur with an R2 of 0.91 for venous blood and 0.81 for finger-prick blood. Compared to FACSCalibur, the PIMA analyzer yielded lower counts with a mean bias of - 47.0 cells/μl (limit of agreement, [LOA]: -204-110 cells/μl) for venous blood and -71.0 cells/μl (LOA: -295-153 cells/μl) for finger-prick blood. For a CD4 threshold of 350 cells/μl, the positive predictive value (PPV) of PIMA was 84.2% and 75.7% and the negative predictive value (NPV) was 97.6% and 95.8% for venous and finger-prick blood, respectively. For an ART threshold of 500 cells/μl, the corresponding PPV was 90.3% and 84.0% and NPV was 94.3% and 93.4%, respectively.

CONCLUSIONSCD4 counting using venous blood with PIMA analyzers is a feasible alternative to a large flow cytometer to determine ART eligibility.

Adolescent ; Adult ; Aged ; Biological Assay ; methods ; Blood Specimen Collection ; CD4 Lymphocyte Count ; methods ; Child ; China ; Female ; HIV Infections ; diagnosis ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult

Previous studies suggested that smoking and passive smoking could increase the risk of breast cancer, but the results were inconsistent, especially for Chinese females. Thus, we systematically searched cohort and case-control studies investigating the associations of active and passive smoking with breast cancer risk among Chinese females in four English databases (PubMed, Embase, ScienceDirect, and Wiley) and three Chinese databases (CNKI, WanFang, and VIP). Fifty-one articles (3 cohort studies and 48 case-control studies) covering 17 provinces of China were finally included in this systematic review. Among Chinese females, there was significant association between passive smoking and this risk of breast cancer [odds ratio (OR): 1.62; 95% confidence interval (CI): 1.39-1.85; I2 = 75.8%, P < 0.001; n = 26] but no significant association between active smoking and the risk of breast cancer (OR: 1.04; 95% CI: 0.89-1.20; I2 = 13.9%, P = 0.248; n = 31). The OR of exposure to husband's smoking and to smoke in the workplace was 1.27 (95% CI: 1.07-1.50) and 1.66 (95% CI: 1.07-2.59), respectively. The OR of light and heavy passive smoking was 1.11 and 1.41, respectively, for women exposed to their husband's smoke (< 20 and ≥ 20 cigarettes per day), and 1.07 and 1.87, respectively, for those exposed to smoke in the workplace (< 300 and ≥ 300 min of exposure per day). These results imply that passive smoking is associated with an increased risk of breast cancer, and the risk seems to increase as the level of passive exposure to smoke increases among Chinese females. Women with passive exposure to smoke in the workplace have a higher risk of breast cancer than those exposed to their husband's smoking.
Breast Neoplasms ; China ; Cohort Studies ; Female ; Humans ; Occupational Exposure ; Odds Ratio ; Risk Factors ; Smoking ; Tobacco Smoke Pollution