OBJECTIVETo investigate the expression of zinc finger E-box binding homebox 1 (ZEB1) in the prepuce of hypospadias children and its relationship to the incidence of hypospadias.

METHODSPrepuce tissues were collected from 37 children aged 6-15 months undergoing hypospadias repair and 11 age-matched controls receiving circumcision. Based on the position of the urethral meatus, the hypospadias cases were classified as severe (n = 13) and mild-moderate (n = 24). The mRNA and protein expressions of ZEB1 were determined by immunohistochemistry and RT-PCR.

RESULTSThe expression of the ZEB1 protein was remarkably higher in the severe (100% [13/13]) and mild-moderate hypospadias patients (75.0% [18/24]) than in the controls (9.1% [1/11]), with statistically significant differences between any two groups (P < 0.05). RT-PCR showed the integrated density value (IDV) of the ZEB1 mRNA expression to be (0.67 ± 0.21), (0.81 ± 0.24), and (1.55 ± 0.29) in the control, mild-moderate, and severe hypospadias patients, respectively, significantly higher in the severe hypospadias than in the control and mild-moderate hypospadias groups (P < 0.05), but with no significant difference between the latter two (P = 0.64).

CONCLUSIONThe expression of ZEB1 is significantly increased in hypospadias patients, and its upregulation is positively correlated with the severity of hypospadias, which suggests that the overexpression of ZEB1 may contribute to the development of hypospadias.

Biomarkers ; metabolism ; Case-Control Studies ; Circumcision, Male ; Foreskin ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Hypospadias ; classification ; etiology ; metabolism ; Immunohistochemistry ; Infant ; Male ; Penis ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism ; Up-Regulation ; Urethra ; Zinc Finger E-box-Binding Homeobox 1

Obstructive sleep apnea (OSA)is a high incidence of potentially dangerous disease,characterized by intermittent hypoxia or hypercapnia.It is an independent risk factor for ischemic stroke.Currently a number of studies have confirmed OSA closely associated with oxidative stress.In this paper,the complex mechanisms of oxidative stress in the OSA and the occurrence of stroke will be reviewed,such as promoting atherosclerosis,damaging the mitochondria,ischemia -reperfusion injury,ischemic preconditioning.To investigate the relationship between OSA,oxidative stress and stroke from molecular mechanisms.

Objective To explore the effects of different vitamin A(VA) levels on thyroid cells apoptosis and its gene expression of mice taking excessive iodine. Methods Kunming mice were randomly divided into 6 groups according to body weight 3 weeks after born: normal control(NI) group, high iodine(HI) group, low vitamin (LVA) group, high iodine plus low vitamin A(HI+LVA) group, high iodine plus vitamin A1 (HI+VA1) group, high iodine plus vitamin A2(HI+VA2) group. The VA was given in food(4000,4000,0,0,8000,16 000 U/kg), and the iodine was given as potassium iodate in water (I-:50,3000,50,3000,3000,3000 μg/L). The apoptosis was tested using in situ end labehng(TUNEL) method. Reverse transcription polymerase chain reaction (RT-PCR) were used to measure the level of mRNA of apoptosis gene(Fas, FasL, Bcl-2) in tissues. Results Apoptotic index measured by TUNEL method was rising along with the mice age. Compared to NI group[(14.09±5.68)%], apoptotic index was significantly increased in HI[(20.91±9.57)%], HI+LVA[(20.29±9.90)%]and HI+VA2 [(19.51±8.25)%]groups in the three months(P < 0.05). Compared to NI group[(16.80±9.90)%], apoptotic index was significantly increased(P < 0.05) in HI[(23.22±8.58)%],LVA[(22.56±6.17)%],HI+LVA [(25.99±9.62)%],HI+VA1 [(21.65±7.74)%]groups in the six months. Compared with the NI group(Fas: 1.29±0.25,1.27±0.26; FasL: 1.60±0.13,1.65±0.13), the mRNA levels of Fas and FasL in HI group(Fas: 1.57±0.36,1.49±0.35; FasL: 1.85±0.46,1.84±0.32) were increased, but the differences were not remarkable(P > 0.05) in the three and six months. Compared with the HI group, the mRNA levels of Fas in HI+ VA1, HI+VA2(1.33±0.35, 1.30±0.26) groups were decreased to the level in NI group in the six months. The mRNA levels of Fas and FasL were not different (P > 0.05) between HI+LVA(I.60±0.27,1.88±0.46) and HI groups in the three months. The mRNA levels of Bcl-2 were not remarkably differences in the three months (1.05±0.19,0.96±0.33,0.95±0.26,1.18±0.27,1.10±0.19,0.98±0.36, all P > 0.05), and in the six months (1.35±0.28,1.60±0.25,1.48±0.18,1.71±0.26,1.66±0.29,1.56±0.35, all P > 0.05). Conclusions Excessive iodine can cause thyroid cells apoptosis in mice. Supplementation of suitable amount of VA can regulate the levels of the apoptosis-related genes expression, and partly antagonize the apoptosis caused by high iodine.

Humans ; Homicide ; Forensic Pathology

OBJECTIVETo review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.

DATA SOURCESThe data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.

STUDY SELECTIONMainly original articles and critical reviews written by major pioneer investigators of this field were selected.

RESULTSThe research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.

CONCLUSIONST4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.

Bacterial Proteins ; metabolism ; DNA-Binding Proteins ; Gene Transfer, Horizontal ; Helicobacter pylori ; genetics ; metabolism ; pathogenicity ; Multigene Family

OBJECTIVETo investigate the mechanism of Panax notoginseng saponins (PNS), an effective component extracted from Panax notoginseng, on atherosclerotic plaque angiogenesis in atherosclerosis-prone apolipoprotein E-knockout (ApoE-KO) mice fed with high-fat, high-cholesterol diet.

METHODSTwenty ApoE-KO mice were divided into two groups, the model group and the PNS group. Ten normal C57BL/6J mice were used as a control group. PNS (60 mg/kg) was orally administered daily for 12 weeks in the PNS group. The ratio of plaque area to vessel area was examined by histological staining. The tissue sample of aortic root was used to detect the CD34 and vascular endothelial growth factor (VEGF) expression areas by immunohistochemistry. The expression of VEGF and nicotinamide adenine dinucleotide phosphate oxidase subunit 4 (NOX4) were measured by reverse transcription polymerase chain reaction and Western blotting respectively.

RESULTSAfter treatment with PNS, the plaque areas were decreased (P<0.05). CD34 expressing areas and VEGF expression areas in plaques were significantly decreased (P<0.05). Meanwhile, VEGF and NOX4 mRNA expression were decreased after treatment with PNS. VEGF and NOX4 protein expression were also decreased by about 72% and 63%, respectively (P<0.01).

CONCLUSIONPNS, which decreases VEGF and NOX4 expression, could alleviate plaque angiogenesis and attenuate atherosclerosis.

Animals ; Down-Regulation ; drug effects ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; NADPH Oxidase 4 ; NADPH Oxidases ; genetics ; metabolism ; Neovascularization, Pathologic ; pathology ; prevention & control ; Panax notoginseng ; chemistry ; Plaque, Atherosclerotic ; pathology ; prevention & control ; Saponins ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo evaluate the antitumor effect of total saponins of R. parvifolius on malignant melanoma.

METHODThe human malignant melanoma A375 cells were regularlly subcultured in vitro, and were divided into five groups contained positive control group (CTX), high concentration (0.01 mg x mL(-1)) and middle concentration (0.001 mg x mL(-1)) and low concentration (0.000 1 mg x mL(-1)) total saponins of R. parvifolius groups and negative control group. By using MTT colorimetric method, the cell viability was measured. B16 melanoma cells were transplanted to mice, which were divided into positive control group, high dose (100 mg x kg(-1)) and middle dose (50 mg x kg(-1)) and low dose (25 mg x kg(-1)) total saponins of R. parvifolius groups and negative control group. The inhibition effect of the tumor in vivo, mean survival time and rate of life-elongation of the mice were observed. TUNEL method was used to detect the apoptosis of B16 malignant melanoma.

RESULTAntitumor assay in vitro showed that the absorbency increased in the concentration of 0.01, 0.001 mg x mL(-1) with statistical significance (P < 0.05 vs negative control). Antitumor assay in vivo showed that the tumor inhibitory rate of high dose (100 mg x kg(-1)) and middle dose (50 mg x kg(-1)) of total saponins of R. parvifolius were 37.02% and 30.61%, respectively. Loaded tumor mouse survival duration could be prolonged. The apoptosis indexes of B16 tumor cells in three treatment groups were 32.5%, 20.5% and 5.5%, respective and there was statistical significance (P < 0.05 vs negative control).

CONCLUSIONThe total saponins of R. parvifolius has remarkable inhibition of proliferation of malignant melanoma cells in vivo and in vitro and exerts antitumor activities through promoting tumor cell apoptosis.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Female ; Humans ; In Situ Nick-End Labeling ; Male ; Melanoma ; pathology ; physiopathology ; Melanoma, Experimental ; pathology ; physiopathology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; Rosaceae ; chemistry ; Saponins ; isolation & purification ; pharmacology

OBJECTIVETo isolate and chondro-inductive culture of human adipose tissue-derived stromal cells and to study their heterotopic chondrogenesis by loading them on alginate gel.

METHODSLiposuction human adipose tissues were minced and digested with collagenase type I. The obtained stromal cells were primarily cultured in BGJb medium for ten days. Secondary harvested cells were cultured in DMEM-F12 medium supplemented with 10%FBS, 6.25 mg/L insulin, 10 mg/L TGF-beta1, 50 mg/L of freshly prepared L-ascorbate for 14 days. After in vitro assay of chondrogenic phenotypes, the cells at density of 10(10)/L were mixed with 1.2% alginate sodium and 102 mmol/L CaCl(2). The cross-linking cell-alginate gel were injected into four BALB/C athymic mice subcutaneously (1 ml for each mouse). Meanwhile, the auto-controls were set by injecting equal dose of simple alginate gel and pure cells in two opposite buttocks of the same mouse subcutaneously. Two mice were sacrificed at fourth and eighth week postoperatively and all samples were removed, fixed, embedded in paraffin and cut into sections of 5 micro m thick. HE staining, Alcian blue and modified Masson's trichrome staining were employed to observe chondrogenesis histologically.

RESULTSAlcian blue and immunocytochemical staining revealed chondroitin sulfate and collagen II in cell matrix after having been chondro-inductive cultured for 14 days. At intervals of fourth and eighth week, heterotopic chondrogenesis is (cartilage formed) within cell-alginate injected sites were found in all mice but negatively in auto-controls. Histologically the hypertrophic chondrocytes were among cartilage matrix in different staining. All alginate gel and solitory cells absorbed within two to three weeks postoperatively in auto-controls.

CONCLUSIONIt seems that stromal cells derived from human adipose tissue presents a potential for chondrogenic differentiation.

Adipose Tissue ; cytology ; Alginates ; pharmacology ; Animals ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Chondrogenesis ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Stem Cell Transplantation ; Stromal Cells ; cytology ; metabolism ; transplantation ; Tissue Engineering

Objective Using the Back Propagation (BP) Neural Network Model to discover the relationship between meteorological factors and mortality of intracerebral hemorrhage,to provide evidence for developing an intracerebral hemorrhage prevention and control program,in Harbin.Methods Based on the characteristics of BP neural network,a neural network Toolbox of MATLAB 7.0 software was used to build Meteorological data of 2007-2009 with intracerebral hemorrhage mortality to predict the effect of BP neural network model,and to compare with the traditional multivariate linear regression model. Results Datas from the multivariate linear regrcssion indicated that the cerebral hemorrhage death mortality had a negative correlation with maximum temperatureand minimum humidity while having a positive correlation with the average relative humidity and the hours of sunshine.The linear correlation coefficient of intracerebral hemorrhage mortality was 0.7854,with mean absolute percentage (MAPE) as 0.21,mean square error (MSE) as 0.22,mean absolute error(MAE) as 0.19.The accuracy of forecasting was 81.31％ with an average error rate as 0.19.The Fitting results of BP neural network model showed that non-linear correlation coefficient of intracerebral hemorrhage mortality was 0.7967,with MAPE as 0.19,MSE as 0.21,MAE as 0.18.The forecasting accuracy was 82.53％ with the average error rate as 0.17.Conclusion The BP neural network model showed a higher forecasting accuracy when compared to the multiple linear regression model on intraccrebral hemorrhage mortality,using the data of 2010' s.

OBJECTIVEThe effects of lentivirus-mediated suppression of Cyclin Y (CCNY) expression on the proliferation of laryngeal cancer cells were investigated in vitro.

METHODSThe lentivirus vectors containing a small hairpin RNA (shRNA) to target CCNY were constructed.Hep-2 cells were divided into the following two experimental groups:the negative control group (control lentivirus infected cells) and CCNY knockdown group (CCNY shRNA-expressing lentivirus infected cells). After Hep-2 cells were infected, Real-time PCR was used to measure CCNY expression. The influence of CCNY on the proliferation of laryngeal cancer cells were assessed using MTT and colony formation experiments.Each experiment was performed in triplicate and repeated three times.

RESULTSLentiviruses expressing shRNA against CCNY were constructed and Hep-2 cells were infected with above mentioned lentivirus at MOI (Multiplicity of infection) of 120.Real-time PCR analysis showed that the mRNA expression of CCNY in Hep-2 cells in the knockdown group was significantly decreased (P < 0.05); the mRNA level of CCNY was 75.3% lower in the si-CCNY group than in the si-CTRL group. After 5 days of lentiviral infection, the cell viability was significantly lower in cells infected with the CCNY-shRNA lentivirus compared to cells infected with the control lentivirus following a 6-day incubation. The colony number was decreased by 60% in Hep-2 cells infected with the CCNY-shRNA-lentivirus infected cells following a 10-day incubation.

CONCLUSIONSThe results suggested that lentivirus-mediated downregulation of CCNY expression decreased the proliferation and growth potency of laryngeal cancer cells.Lentiviruses delivering shRNA against CCNY may be a promising tool for laryngeal cancer therapy.

Cell Line, Tumor ; Cell Proliferation ; Cyclins ; Humans ; Laryngeal Neoplasms ; metabolism ; Lentivirus ; genetics ; RNA, Small Interfering ; genetics