4 weeks of arrested embryos.The AAH expression was found to have the similar result as HIF-1?'s.Conclusions The expression level of HIF-1? and AAH in villi of missed abortion patients is much lower than that of normal early pregnant women.HIF-1? and AAH have a function of supporting normal pregnancy,so their low expression may be an important cause of missed abortion.

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

OBJECTIVE To enhance the rational usage of antibiotics by comprehensive interventional measures in clinics.METHODS Several interventional measures have been adopted in our hospital since January 2001: to(establish) expert team on antibiotics usage and administration consultation;constitute antibiotics use criteria(suitable) for each clinical specialty;train and examine the usage of antibiotics;censor the distribution of pathogen and drug-resistance variance.Then 10% of the discharged medical records in 2000,2002 and 2004 were drawn out respectively to analyze the usage of antibiotics and the isolation of pathogen from nosocomial infection cases.(RESULTS) The proportion of the patients with prophylactic and remedial indications was increased remarkably((P

Objective To discuss function of the fusion cells of human bone marrow stromal cells (BMSCs) and human myeloma cell RPMI 8226 in the pathogenesis of multiple myeloma bone disease.Methods The cells,labeled by cell tracer green fluorescent probe (CMFDA) and red fluorescent probe (CMTMR),respectively,were induced into fusion by chemical polyethylene glycol (polyethyleneglycol,PEG-1000),and cell fusion model was set up.Whether fusion cells had nucleus fusion was determined by Karyotype analysis.The expressions of stemness-related genes,SIRP α gene and DC-STAMP gene in fusion cells were identified.Results Polyethylene glycol (PEG-1000) could mediate the integration of BMSCs and RPMI 8226 cells.The number of chromosomes in more than 80 ％ the hybrid cells was about 80.Fusion cells not only showed that BMSCs,stemness-related genes of c-myc,Klf-4 and OCT-4 genes expressed positively,but also the fusion-related genes SIRPα and DC-STAMP expressed positively.Conclusion BMSCs and RPMI 8226 cells can form fusion cells,and the cells have the potential for further integration,which is one of the important reasons for the promotion of muhiple myeloma bone destruction.

Objective： To elucidate the role of bone marrow stromal cells in cooperation with exogenous cytokines in hematopoiesis. Methods： Fetal bone marrow stromal cells (FBMSC) was combined with cytokines including SCF,IL-3,IL-6,GM-CSF in a 5-day liquid culture system of adult bone marrow mononuclear cells, then we cultured bone marrow derived CD34+-enriched cells with FBMSC+SCF+IL-3+IL-6+G-CSF+EPO for 2 weeks. Results：FBMSC were in good cooperation with above mentioned exogenous cytokines. When CD34+-enriched cells from adult bone marrow were cultured with combinations of FBMSC, SCF, IL-3, IL-6, G-CSF and EPO, total nucleated cells, CFU-GM, BFU-E and CD34+ cells were increased by 119.6±30.9, 54.6±17.4, 25.2±4.4, 11.1±4.2 folds, respectively. Conclusion：FBMSC in cooperation with exogenous cytokines support the in vitro expansion of human hematopoietic progenitor cells efficiently.

OBJECTIVETo evaluate the effect of Qufeng Zhidong Recipe (QZR) on the head tic behavior, and the mRNA expressions of Notch1 and dopamine D2 receptor (D2R) in tic disordered mice.

METHODSMouse model like wet-dog shake head tic disorder was established by peritoneal injection of 5-HT2A/C agonist DOI for 14 successive days. The model mice were divided into three groups, the model group, the Chinese medicine (CM) treated group and the Western medicine (WM) treated group, they were intervened respectively with distilled water, QZR (10 g/kg) and haloperidol (1 mg/kg). Besides, a normal control group was set up and gastrogavaged with distilled water. The effect of intervention was evaluated 2 weeks later by estimating the head tic and the creeping distance of animals, and the mRNA expressions of D2R and Notch1 in corpus striatum and prefrontal cortex regions were detected using Real-time PCR.

RESULTSThe wet-dog shake response and the creeping distance of mice were significantly reduced after intervention in both intervened groups, showing insignificant difference between the effects of CM and WM (P > 0.05). The expression of D2R mRNA in corpus striatum was higher than that in the prefrontal cortex (P < 0.01), at the prefrontal cortex, it was 151 +/- 30 in the CM group and 180 +/- 41 in the WM group, and at the corpus striatum, 710 +/- 64 and 850 +/- 80 respectively, all higher than those in the model group (P < 0.05). While the Notch1 mRNA expression in model mice were lower at the prefrontal cortex than at the corpus striatum (P < 0.05). After intervention it was 55 +/- 20 in the CM group and 48 +/- 23 in the WM group at the prefrontal cortex, all significantly lower than that in the model group (P < 0.05).

CONCLUSIONDOI-induced wet-dog shake response could well simulate the clinical characteristics of tic disorder; QZR could improve the tic behavior and creeping distance in the model mice. The up-regulation of D2R mRNA expression after QZR intervention may be related with the down-regulation of Notch1 expression, this findings is worthy of further studies.

Animals ; Corpus Striatum ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Receptor, Notch1 ; genetics ; metabolism ; Receptors, Dopamine D2 ; genetics ; metabolism ; Signal Transduction ; drug effects ; Tic Disorders ; drug therapy

Chemoembolization, Therapeutic ; adverse effects ; methods ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Kidney Neoplasms ; therapy ; Male ; Prospective Studies ; Wilms Tumor ; therapy

Objective： To elucidate the role of bone marrow stromal cells in cooperation with exogenous cytokines in hematopoiesis. Methods： Fetal bone marrow stromal cells (FBMSC) was combined with cytokines including SCF,IL-3,IL-6,GM-CSF in a 5-day liquid culture system of adult bone marrow mononuclear cells, then we cultured bone marrow derived CD34+-enriched cells with FBMSC+SCF+IL-3+IL-6+G-CSF+EPO for 2 weeks. Results：FBMSC were in good cooperation with above mentioned exogenous cytokines. When CD34+-enriched cells from adult bone marrow were cultured with combinations of FBMSC, SCF, IL-3, IL-6, G-CSF and EPO, total nucleated cells, CFU-GM, BFU-E and CD34+ cells were increased by 119.6±30.9, 54.6±17.4, 25.2±4.4, 11.1±4.2 folds, respectively. Conclusion：FBMSC in cooperation with exogenous cytokines support the in vitro expansion of human hematopoietic progenitor cells efficiently.

Hepatocyte growth factor (HGF) pretreatment could protect multiple cell types from apoptosis induced by various damages including oxidative stress. The present study was designed to investigate the protective effect of HGF on rat cortical neurons against apoptosis induced by hydrogen peroxide (H2O2) in culture, and then to explore whether HGF could influence the mitochondrial pathway of apoptosis. Primary rat cortical neurons were isolated from Sprague-Dawley rats and cultured in serum free medium containing 2% B27 and Neurobasal-A. To mimic the oxidative stress damage, cortical neurons were exposed to 100 mumol/L H2O2 for 4 h. To explore the effects of HGF on the neurons subjected to H2O2 injury, cells were pretreated with HGF 15, 30, 60 ng/mL for 24 h, respectively, and then exposed to 100 mumol/L H2O2 for 4 h. The cell viability was measured by MTT colorimetric assay and cell injury was evaluated by lactate dehydrogenase (LDH) leakage rate. Apoptotic cells were detected by Hoechst 33258 staining and Annexin V-FITC/PI double labeled flow cytometry. The caspase-3 activity was assessed by colorimetry. The alteration of transmembrane potential of mitochondria was determined by confocal laser scanning microscopy. The expression of cytochrome C protein was measured by Western blot analysis. The results showed that H2O2 treatment significantly decreased the cell viability, increased LDH leakage rate and the percentage of apoptotic cells. Pretreatment of HGF at different concentrations (15-60 ng/mL) could remarkably increase the cell viability of neurons. Compared with that of H2O2 group (53.4%+/-7.4%), the cell viabilities of neurons treated with 15, 30, and 60 ng/mL HGF significantly increased to (69.3+/-6.4)%, (77.5+/-6.1)% and (82.9+/-9.3)% (P<0.05), respectively. HGF preincubation also evidently decreased the LDH leakage rate in cortical neurons damaged by H2O2. The results of Hoechst staining revealed that HGF pretreatment could significantly reduce the apoptotic rate of neurons. The apoptotic rate of H2O2 group was (62.8+/-7.1)%, while that of HGF groups decreased significantly to (34.8+/-8.4)%, (23.5+/-3.2)% and (18.6+/-4.5)% (P<0.05), respectively. The data from caspase-3 activity assay indicated that HGF preconditioning could also remarkably decrease the caspase-3 activity of neurons. In addition, in the presence of various concentrations of HGF, the decrease of transmembrane potential of mitochondria in neurons caused by H2O2 injury could be reversed. Moreover, as detected by Western blot analysis, HGF downregulated the expression of cytochrome C protein in neurons. These results suggest that HGF has a protective effect on rat cortical neurons against apoptosis induced by H2O2, which might be related to the inhibition of the mitochondrial apoptotic pathway and the suppression of the caspase-3 activity.
Animals ; Apoptosis ; Brain ; cytology ; Caspase 3 ; metabolism ; Cell Survival ; Cells, Cultured ; Cytochromes c ; metabolism ; Hepatocyte Growth Factor ; pharmacology ; Hydrogen Peroxide ; pharmacology ; Mitochondria ; physiology ; Neurons ; cytology ; drug effects ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the genetic characteristics of measles viruses circulating in Zhejiang province in 2005.

METHODS4 groups of measles viruses isolated in outbreaks and the H and N gene were amplified by reverse transcription polymerase chain reaction (RT-PCR). PCR products were purified, sequenced and data was analyzed.

RESULTSAll of the 4 measles isolates belonged to genotype H1 which had been a main genotype containing all of the isolates in China. The isolates shared 99.2% -99.7% identity of amino acid sequence on H and 99.8% identity of amino acid sequence on N gene. When comparing to the China vaccine strain (Shanghai 191), there were 95.2%-95.5% homogeneties and 95.5% homogeneties on H and N gene respectively.

CONCLUSIONData from phylogenic trees of H and N gene revealed that the wide-type measles viruses circulating in Zhejiang province in 2005 all belonged to genotype H1. There were obvious differences on genetic characteristics between the isolates and the genotype A (Shanghai 191).

Amino Acid Sequence ; China ; epidemiology ; Genes, Viral ; Genotype ; Measles ; epidemiology ; Measles virus ; classification ; genetics ; isolation & purification ; Phylogeny