OBJECTIVETo investigate the methylation of p16(INK4a) and RB gene, and the expression of p16(INK4a) in meningiomas.

METHODSMethylation-specific polymerase chain reaction (MSP) was used to detect the methylation of p16(INK4a) and RB in 50 cases of meningiomas, and immunostaining was performed to analyze the protein expression of p16(INK4a) in 25 of those cases.

RESULTSNo methylation was found in the benign meningiomas, whereas methylation of p16(INK4a)or RB occurred in 6(37.5%) cases of grade II tumors and 4(28.6%) cases of grade III tumors, and among these cases, an atypical meningioma showed methylation of both genes. Thirteen cases showed p16(INK4a) positive expression, but none of them was methylated.

CONCLUSIONThe methylation of p16(INK4a) or RB is related with the tumorigenesis and progression of atypical and anaplastic meningiomas, and a probable mechanism is that methylation causes the loss of expression and leads to dysfuncation of the p16(INK4a)/cyclin D1/CDK4/RB pathway.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cyclin D1 ; genetics ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinases ; genetics ; DNA Methylation ; Female ; Genes, Retinoblastoma ; Genes, p16 ; Humans ; Male ; Meningeal Neoplasms ; genetics ; Meningioma ; genetics ; Middle Aged ; Proto-Oncogene Proteins

OBJECTIVETo investigate the influence of down-regulating Smoothened (SMO) gene expression through short hairpin RNA (shRNA) on the proliferation of breast cancer stem cells.

METHODSHuman SMO shRNA was designed, synthesized chemically, and transfected into MCF-7 cells to down-regulate SMO gene. By using G418, stable cells with down-regulated SMO were selected. In vitro proliferation of these cells was measured by CCK8 assay. The proportion of CD44(+)/CD24(-) cells was detected by flow cytometry and the mammospheres formation was determined by suspension sphere culture. The expression of SMO, GLI1 and Oct4 was detected by Western blot. In vivo, the volume of tumor was measured every 3 days and the expression of SMO, GLI1 and Oct4 detected by Western blot.

RESULTSIn vitro, the cells were transfected with SMO-shRNA and selected by G418 after 21 days. SMO-shRNA effectively down-regulated the expression of SMO gene and protein, and inhibited the proliferation of MCF-7 and markedly reduced the proportion of CD44(+)/CD24(-) cells and mammospheres. In vivo, SMO-shRNA treatment of MCF-7 significantly inhibited the volume of tumor. The positive rate of SMO in negative control and SMO-shRNA group was 5/5 and 2/5, respectively. The expression of SMO, GLI1 and Oct4 in different groups were 0.72 ± 0.17 and 0.21 ± 0.09, 1.21 ± 0.21 and 0.47 ± 0.12, 0.83 ± 0.13 and 0.25 ± 0.07. SMO, GLI1 and Oct4 down-regulation significantly suppressed at protein levels (P < 0.05).

CONCLUSIONThe shRNA by chemical synthesis can effectively down-regulate SMO gene expression and inhibit the proliferation of breast cancer stem cells.

Animals ; Cell Proliferation ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hyaluronan Receptors ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; pathology ; Octamer Transcription Factor-3 ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Smoothened Receptor ; Transcription Factors ; metabolism ; Transfection ; Tumor Burden ; Zinc Finger Protein GLI1

OBJECTIVETo assess the reliability and validity of the Aging Males' Symptoms (AMS) scale in the male population of Shanghai.

METHODSWe enrolled 973 males aged 40 years and over in a community of Shanghai, China. Using the AMS scale, we calculated the split-half reliability coefficient and Cronbach's alpha coefficient, assessed the validity through confirmatory factor analysis and correlation analysis, and obtained the domain scores of different people by analysis of variance and independent sample test.

RESULTSThe split-half reliability was > 0.78 (P < 0.01) and Cronbach's alpha coefficients of all the dimensions > 0.82 (P < 0.01). Confirmatory factor analysis showed 3 domains in the AMS scale, Pearson correlation coefficients of all the items to their domains were > 0.49 (P < 0.01), and the total testosterone level was not correlated with AMS scores, with Pearson correlation coefficient of -0.04 (P > 0.05). Statistically significant differences were found in AMS scores among different age groups as well as among those with different chronic disease histories, but not in the psychological domain among different age groups.

CONCLUSIONThe reliability and validity of the AMS scale are acceptable in assessing aging males'symptoms among the male population of Shanghai, but further studies are needed to determine whether it could be used as a tool for screening late-onset hypogonadism (LOH) in males.

Adult ; Aged ; Aging ; China ; epidemiology ; Humans ; Hypogonadism ; epidemiology ; psychology ; Male ; Middle Aged ; Psychometrics ; Reproducibility of Results ; Surveys and Questionnaires

Objective To investigate the diagnostic criteria for HBV-related acute-on-chronic pre-liver failure (pre-ACLF) which can effectively predict the risk of liver failure.Methods A total of 1279 patients with severe icteric chronic hepatitis B (CHB) and/or severe acute exacerbation of CHB were enrolled.The influence of serum levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),and total bilirubin (TBil),international normalized ratio (INR) of prothrombin time,sex,and age on the incidence rate of acute-on-chronic liver failure (ACLF) was analyzed,the diagnostic criteria for pre-ACLF and predictive model for ACLF were developed.The chi-square test was used for comparison of categorical variables,and the independent samples t-test was used for continuous data;multivariate logistic regression analysis was performed to evaluate the risk of liver failure.Results The baseline serum levels ofALT,AST,and TBil,and INR were independent risk factors for liver failure (P ＜ 0.05).The diagnostic criteria for pre-ACLF were as follows:(1) INR ≥ 1.30;(2) AST ≥ 10×upper limit of normal (ULN) and obvious jaundice (TBil ≥ 51.3 μmol/L),or TBil ≥ 342.0 μmol/L.These criteria had a positive predictive value of 45.9％,a negative predictive value of 89.8％,a sensitivity of 69.1％,and a specificity of 76.9％.The predictive model for the risk of ACLF was PY =1=eX/(1+ex) (PY represented positive results of logistic regression analysis),X =-10.245+0.026×AST(ULN)-0.025×AST(ULN)+0.046×TBil(mg/d1) + 4.642×INR+0.049×age(years).The patients with higher PY values tended to have a higher incidence rate of ACLE The incidence rate of ACLF was 75.3％ in patients with PY ≥ 0.60,more than 50％ in patients with a PY value of 0.40-0.59,and 1.8％ in patients with PY ＜ 0.10 (P ＜ 0.01).Conclusion The diagnostic criteria for pre-ACLF and predictive model can effectively evaluate the risk of HBV-related ACLF.

OBJECTIVETo investigate the pathogenesis and treatment of penile necrosis resulting from microwave diathermy following circumcision.

METHODSWe retrospectively analyzed the clinical data about 9 cases of penile necrosis resulting from postoperative microwave diathermy following circumcision. The 9 males, aged 20 - 39 (mean 26) years, underwent traditional circumcision for redundant prepuce or phimosis in other hospitals, followed by microwave diathermy for 30 - 60 minutes daily, which resulted in penile necrosis. With no response to conservative therapy, the patients were referred to our hospital at 3 -30 days postoperatively. Of the 9 patients, 5 presented with dry gangrene and 4 with moist gangrene. Six of the patients underwent partial penectomy, including 1 that received penis lengthening.3 months later, while the other 3 underwent total penectomy for total penile necrosis followed by penile reconstruction 3 months later, with deep inferior epigastric perforator (DIEP) flaps and by implantation of the 12th costal cartilage in 2 cases and with epigastric groin island flaps and by urethroplasty in the other.

RESULTSThe patients were followed up for 2 - 8 years, and all could urinate smoothly in the standing position. Of the 6 men treated by partial penectomy, 1 received penis lengthening and achieved a penile length of 7 cm and 5 had the remaining penile length of 3 -5 cm, 4 with erectile function and the other 2 capable of sexual intercourse. The 3 men treated by total penectomy achieved nearly normal external appearance of the penis, with a finalized length of (11.7 ± 1.3) cm, a circumference of (11.4 ± 2.1) cm, and a normal feel of the skin. Of the 3 cases of penile reconstruction, 2 achieved sufficient erectile hardness of the penis (grade 3) for sexual intercourse, while the other 1 remained impotent.

CONCLUSIONPost-circumcision microwave diathermy may result in penile necrosis, for the management of which, early debridement is necessitated and penile lengthening or reconstruction can be performed according to the severity of the lesion and needs of the patient.

Adult ; Circumcision, Male ; methods ; Coitus ; Costal Cartilage ; transplantation ; Diathermy ; adverse effects ; methods ; Humans ; Male ; Microwaves ; adverse effects ; Penis ; abnormalities ; surgery ; Phimosis ; surgery ; Postoperative Period ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Young Adult

OBJECTIVETo explore the therapeutic methods, surgical indications and clinical practice of minimally invasive surgery on traumatic epidural hematoma (EDH).

METHODSRetrospective study was made on 135 patients with traumatic EDH admitted into our hospital from June 2002 to August 2005. Sixty-five patients were treated with mini-invasive negative pressure drainage (treatment group), 70 patients with comparable condition used traditional craniotomy (control group). The mean time of operation, average days in hospital, expenditure and prognosis of two groups were recorded and analyzed.

RESULTSThere was no significant difference in therapeutic efficacy between two groups. Patients in treatment group had a shorter hospital stay and less expenditure than those in control group.

CONCLUSIONMini-invasive negative pressure drainage is simple, effective, economical and applicable to some traumatic EDH patients.

Brain Injuries ; complications ; Female ; Hematoma, Epidural, Cranial ; surgery ; Humans ; Male ; Minimally Invasive Surgical Procedures ; Retrospective Studies

The study was purposed to investigate the immune regulatory effects of human bone marrow mesenchymal stem cells (hMSCs) on Foxp3 expressing CD4(+)CD25(+) regulatory T cells and to explore the mechanism of immune modulation by hMSCs. Human MSCs were isolated and expanded from bone marrow cells, and identified with cell morphology, and the phenotypes were assessed by immunohistochemistry. Human peripheral blood mononuclear cells (hPBMNCs) were prepared by centrifugation on a Ficoll Hypaque density gradient. The hMSCs (1 x 10(3), 1 x 10(4), 1 x 10(5)) were added into wells containing hPBMNCs (1 x 10(6)) from an unrelated donor in the presence of rhIL-2. After 5 days of co-culture, the percentage of CD4(+)CD25(+) T cells was detected by flow cytometry. T cell proliferation was assessed by [(3)H] thymidine incorporation using a liquid scintillation counter. The expression of Foxp3 in CD4(+)CD25(+) T cells was detected by reverse transcription polymerase chain reaction (RT-PCR). Cytokines (TGF-beta, IL-12, IFN-gamma, IL-10) concertrations of cultured supernatants were measured with ELISA. The results indicated that in all the experiments, the presence of hMSCs with hPBMNCs resulted in a statistically significant decrease in T cell proliferation, in dose-dependent manner. The increase of percentage of CD4(+)CD25(+) T cells in the peripheral CD4(+) T cell was observed after coculturing lymphocytes with hMSCs (p < 0.01). The expression of Foxp3-mRNA (Foxp3/beta-actin) in hMSCs groups was significantly higher than that in the control and was negatively associated with the value of CPM representing T proliferation. The levels of TGF-beta and IL-10 were higher in hMSCs groups than that in the control, and the levels of TGF-beta and IL-10 correlated positively with Foxp3-mRNA expression and the percentage of CD4(+)CD25(+) T cells. However, the secretion of IL-12 and IFN-gamma was significantly attenuated by hMSCs coculture, and there was no correlation with Foxp3-mRNA expression and the percentage of CD4(+)CD25(+) T cells. It is concluded that the Foxp3 expressing regulatory T cells may play an important role in the immune regulatory by hMSCs. Its mechanism is related to that the hMSCs-mediated TGF-beta and IL-10 convert CD4(+)CD25(-) T cells into CD4(+)CD25(+) T regulatory T cells, which specifically inhibits the proliferation of T cells.
Bone Marrow Cells ; cytology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Forkhead Transcription Factors ; metabolism ; Humans ; Immunization ; Interleukin-10 ; biosynthesis ; Interleukin-2 Receptor alpha Subunit ; immunology ; metabolism ; Mesenchymal Stromal Cells ; cytology ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Transforming Growth Factor beta ; biosynthesis

OBJECTIVETo observe the effects of complement fragment C3f on expression and secretion of collagen I, III and transforming growth factor( TGF)-beta1 in human embryonic lung fibroblast (MRC-5) cells.

METHODSMRC-5 cells were cultured with C3f (the synthetic 17 peptides fragments of complement C3). The extracellular and intracellular expression levels of type I, III collagens and TGF-beta1 in MRC-5 cultures were detected by ELISA and immunohistochemistry, respectively.

RESULTSThe expression levels of type I, III collagen and TGF-beta1 in the supernatant of MRC-5 cultures decreased significantly with the concentrations of C3f as compared with controls (P < 0.05). Also the expression level of TGF-beta1 in MRC-5 cytoplasm reduced significantly as compared with controls (P < 0.05).

CONCLUSIONThe results of present in vitro study showed that the complement fragment C3f could reduce the formation of TGF-beta1 and type I, III collagens in MRC-5 cells, and inhibit the lung tissue fibrosis.

Cell Line ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Complement C3b ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Humans ; Lung ; cytology ; drug effects ; embryology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo study the effects of behind legs vibrations on erectile function in male rabbits through the concentration of plasma reproductive hormone and the expression of nitric oxide synthase (eNOS), endothelin-1 (ET-1) mRNA in vibrated male rabbits.

METHODS30 male adult rabbits were assigned randomly to A group (vibration power: 3.02 m/s(2)), B group (vibration power: 6.13 m/s(2)), C group (vibration power: 12.26 m/s(2)) and control group. The concentration of expression of eNOS, ET-1 mRNA were measured with RT-PCR after rated for 30 days.

RESULTS(1) Compared with 0 days vibration, after exposure to vibration for 10, 20, 30 days, the A, B, C group concentration of plasma T, LH are much lower (P < 0.05), the concentration of plasma E2 is much higher. (2) Compared with control group after exposed for 30 days, the expression of ET-1 mRNA [B group:(17.39 +/- 4.59) x 104; C group: (36.21 +/- 13.13) x 104 ] were much higher and expression of eNOS mRNA [A group: (19.11 +/- 6.83) x 104; B group: (11.86 +/- 3.15) x 104; C group: (4.68 +/- 3.26) x 104] was much lower, there were significant differences (P < 0.01).

CONCLUSIONSThe vibration of behind legs in rabbits resulted the concentration of plasma T, LH are much lower, the concentration of plasma E2 is much higher, increased the expression of eNOS mRNA, decreased the expression of eNOS mRNA, then vary the erectile function.

Animals ; Endothelin-1 ; genetics ; metabolism ; Male ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; Penile Erection ; Penis ; metabolism ; RNA, Messenger ; genetics ; Rabbits ; Vibration ; adverse effects

Objective To study the mechanism of IL-6 stimulated the secretion of IL-10 in nasopharyngeal carcinoma CNE-2 cells.Methods Nasopharyngeal carcinoma CNE-2 cells were cultured with recombination cytokine IL-6 in vitro,or anti-IL-6 receptor antibody and signal pathway inhibitor were pre-incubated for 1 hour,and then IL-6 were added,the mRNA and protein level of IL-10 were separately detected by RT-PCR and enzyme-linked immunosorbent assay.Results Compared with the control group,the expression and secretion of IL-10 in IL-6 stimulated group were significantly increased,which was in a dose and time dependent way,the difference was significant(P ＜ 0.01).Additionally,IL-6 stimulated the expression and secretion of IL-10 by CNE-2 cells were significantly decreased in following pre-incubated with anti-IL-6 receptor antibody or NF-κB inhibitor,the difference was significant(P ＜ 0.01),but such effect was not detected when CNE-2 cells were pre-incubated with the PI3K inhibitor,p38/MAPK inhibitor,JNK inhibitor,MEK1/2 inhibitor and STAT3 inhibitor.Conclusion IL-6 can induce the expression and secretion of IL-10 in nasopharyngeal carcinoma CNE-2 cells via IL-6R/NF-κB signal pathway,and blocking IL-6 signal may be useful for the immunotherapy of nasopharyngeal carcinoma.