OBJECTIVE: To develop a novel technique of optical recording and its validation for assessment of the neuroprotective effect of nimodipine, a L-type calcium channel blocker. METHODS: In vitro ischemia was induced by oxygen/glucose deprivation (OGD), the light transmittance (LT) of rat hippocampal slices undergoing OGD and reperfusion was quantitated using a simple apparatus relying on basic principles of light transmittance and a computerised image analysis system. RESULTS: OGD was associated with increased LT in the stratum radiatum of CA1 area and the dentate gyrus in hippocampal slices. Peak LT occurred (7.59 +/-1.42) min after OGD, followed by a marked decrease in LT (n=15 slices). Nimodipine administration (0.5 &mgr;mol/L, n=10 slices, 5 &mgr;mol/L, n=9 slices) appeared to protect the tissue from OGD damage by inhibiting elevation of LT, However, 50 &mgr;mol/L nimodipine resulted in increased LT (25.83 +/-6.32). min after administration (n=11 slices). CONCLUSION: LT signal measurement is a non-invasive, reliable method for determination of neuronal damage in ischemic rat brain slices Nimodipine is demonstrated opposite neuroprotective effects depending on its dose.

Objective: To study the recombinant epitope antigens of hepatitis C virus (HCV), in order to fulfil the requirements of recombinant immunoblot assay kit. Methods: An expressing vector pBVIL1 for expression of recombinant antigens in a fusion manner with IL-1β was constructed. A series of selected genes from the HCV antigens including the C, NS3, NS4 and NS5 were amplified from HCV gene-containing plasmids using PCR and the expression plasmids for these genes were constructed in pBVIL1, respectively. The activity of the purified recombinant antigens were tested against an identified HCV antibody positive and negative panel with ELISA. Results and Conclusions: All the cloned genes of chosen antigen epitopes were highly expressed in pBVIL1 in E.coli. The activity of the C and NS4 antigens were slightly higher than the RIBA3.0 antigens, while the activity of NS3 was slightly lower than the RIBA3.0 antigen. But the total evaluation for the panel was same as RIBA3.0. That means the cloned antigens were suitable for the use in RIBA test kit.

AIMTo determine the effect of histamine on ischemia-induced cellular edema and viability reduction in rat hippocampal slices, and the involved subtypes of histamine receptor in this effect.

METHODSIn vitro ischemic injury of hippocampal slices was induced by oxygen-glucose deprivation (OGD). The slice injury was determined by real-timely measuring the changes of light transmittance (LT) for the cellular edema in CA1 region of the hippocampal slice, and by detecting the product of 2, 3, 5-triphenyltetrazolium chloride (TTC), formazan, for the slice viability. The effect of histamine at various concentrations on the slice injury was observed, and the blockage by antagonists of histamine receptors was also investigated.

RESULTSHistamine (0.01-10 micromol x L(-1)) inhibited the peak value of LT during OGD in hippocampal slices and improved the reduced viability after OGD. Diphenhydramine (0.1-10 micromol x L(-1)), an H1 receptor antagonist, did not affect the effect of histamine, while cimetidine (0.1-10 micromol x L(-1)), an H2 receptor antagonist, partly abolished the protective effect of histamine.

CONCLUSIONHistamine protects hippocampal slices against ischemia-induced cellular edema and viability reduction; this effect might be mediated via, at least partly, H2 receptor.

Animals ; Cell Hypoxia ; Cell Survival ; drug effects ; Cimetidine ; pharmacology ; Diphenhydramine ; pharmacology ; Formazans ; metabolism ; Glucose ; deficiency ; Hippocampus ; drug effects ; metabolism ; pathology ; Histamine ; pharmacology ; Histamine H1 Antagonists ; pharmacology ; Histamine H2 Antagonists ; pharmacology ; Male ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the genetic etiology of an autosomal dominant dentinogenesis imperfecta in a Chinese family.

METHODSThe molecular change of the disease in the family was analyzed through the clinical examination, linkage analysis, mutational screening of the DSPP gene and restriction fragment length polymorphism analysis.

RESULTSThe disease related gene was completely linked with microsatellite marker D4S1534. We found a novel mutation in the first exon of the DSPP gene (c.49C>T, p.Pro17Ser). All patients in the family had the mutation, while this mutation was not observed in the normal individuals of this family and 100 unrelated controls.

CONCLUSIONThe p.Pro17Ser identified in the family was a new pathogenic mutation. Our finding provided further understanding of the molecular mechanism of dentinogenesis imperfecta.

Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Dentinogenesis Imperfecta ; genetics ; Exons ; Extracellular Matrix Proteins ; genetics ; Female ; Humans ; Male ; Microsatellite Repeats ; Molecular Sequence Data ; Mutation ; Pedigree ; Phosphoproteins ; Sialoglycoproteins ; Young Adult

OBJECTIVETo develop oxygen/glucose deprivation (OGD)-and NMDA-induced neurotoxicity models in rat primary neurons and hippocampal slices, and to determine the protective effect of minocycline.

METHODSThe injuries of primary neurons were induced by OGD or NMDA (50micromol/L). Morphological changes of neurons were observed, and neuron viability was evaluated by MTT assay. The changes of light transmittance (LT) were induced by OGD or NMDA in rat hippocampal slices. The effects of minocycline and MK-801, an NMDA receptor antagonist, were observed in the models of OGD-or NMDA-induced injuries.

RESULTMinocycline concentration dependently inhibited OGD induced decrease of neuron viability and ameliorated neuron morphological changes at 1 and 10 micromol/L. It also inhibited NMDA insult at 10 and 100 micromol/L. MK-801 inhibited both injuries at 1 micromol/L. However, minocycline at 1 or 10 micromol/L did not inhibit the augment of LT in hippocampal slices induced by OGD or NMDA, while MK-801 inhibited both OGD-and NMDA-induced LT augments.

CONCLUSIONMinocycline protects neurons from OGD insult, which may inhibit NMDA receptor-mediated neurotoxicity through an indirect pathway, but has no effect on OGD-or NMDA-induced immediate injury in hippocampal slices.

Animals ; Cell Hypoxia ; Cells, Cultured ; Dizocilpine Maleate ; pharmacology ; Glucose ; metabolism ; Hippocampus ; drug effects ; Male ; Minocycline ; pharmacology ; N-Methylaspartate ; toxicity ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

UNLABELLEDTo investigate the relationship between Ureaplasma urealyticum (UU) infection and semen quality.

METHODSFrom 2001 to 2003, 346 eligible patients aged 20-45 years were invited from two hospitals in Shanghai, China, to participate in an investigation which included questionnaires about general and reproductive health, an external genital tract examination, UU culture and semen analysis. Multiple linear regression models were used to examine whether UU had a significant effect on semen quality after adjustment for confounding factors.

RESULTSFindings suggested that UU infection was associated with higher semen viscosity and lower semen pH value. Sperm concentration was lower in UU positive subjects than that in UU negative subjects (54.04 X 10(6)/mL vs.70.58 X 10(6)/mL). However, UU did not significantly affect other semen quality indexes.

CONCLUSIONUU infection of the male genital tract could negatively influence semen quality.

Adult ; Clothing ; Humans ; Infertility, Male ; epidemiology ; Male ; Middle Aged ; Physical Examination ; Semen ; physiology ; Sperm Count ; Sperm Motility ; Spermatozoa ; physiology ; Surveys and Questionnaires ; Testis ; anatomy & histology ; Ureaplasma Infections ; physiopathology ; Ureaplasma urealyticum

OBJECTIVETo analyze treatment and prognostic factors of 74 patients with dermatofibro-sarcoma protuberans (DFSP).

METHODSFrom August 1990 to November 1999, 74 patients with DFSP confirmed pathologically were treated. There were 52 males and 22 females with a median age of 37 years (range 4 to 80 years) on diagnosis. Seventeen patients were treated by extensive excision and 2 by limited excision. Fifty-two patients had surgical resection alone (S), and 22 postoperative radiotherapy (S + R) of 50-70 Gy. The multivariate parameters were analyzed using Cox model. Kaplan-Meier and Log-Rank test were used to evaluate the results of the recurrence-free survival.

RESULTSThe rate of recurrence was 28.4% for all patients. The 5-year recurrence-free survival rate (RFSR) was 66.6% and the 10-year RFSR was 52.5%. The 5-year and 10-year in the S group were 58.4% and 41.2%, compared with 90.0% and 83.3% in the S + R group (P < 0.05). The 5-year and 10-year RFSR in the pathologically positive margin group were 57.5% and 41.4% respectively, compared with the 75.0% and 56.6% in the pathologically negative group (P < 0.05). Multivariate analysis suggested radiotherapy and negative pathological margins were favorable prognostic factors.

CONCLUSIONPost-operation radiotherapy and pathological margin are the independent prognostic factors.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Combined Modality Therapy ; Dermatofibrosarcoma ; mortality ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Postoperative Care ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Skin Neoplasms ; mortality ; radiotherapy ; surgery

OBJECTIVETo investigate the outcome of in vitro fertilization and embryo transfer (IVF-ET) in couples with the husband positive for chronic infection of hepatitis B virus (HBV).

METHODSThis study involved 102 infertile couples receiving IVF-ET with the husbands(but not the wives) positive for hepatitis B surface antigen (HBsAg), and another 204 couples negative for HBsAg receiving the treatment served as the control group. The cumulative embryo score, fertilization rate, cleavage rate, rate of good quality embryos, implantation rate, clinical pregnancy rate, first trimester and late miscarriage rates, delivery rate, and neonatal malformation rate were recorded and compared between the two groups.

RESULTSBetween the HBsAg-positive and the control groups, the cumulative embryo score (52.8-/+18.7 vs 55.4-/+16.9), insemination rate (66.9% vs 66.1%), cleavage rate (97.6% vs 97.2%), rate of good quality embryos (34.0% vs 37.1%), implantation rate (40.9% vs 34.6%), clinical pregnancy rate (56.9% vs 50%), first trimester miscarriage rate (6.9% vs 5.9%) and late pregnancy miscarriage rate (8.6% vs 4.9%), delivery rate (40.2% vs 43.6%) and neonatal malformation rate (0 vs 0) were all similar (P>0.05;).

CONCLUSIONChronic HBV infection in the husband might not affect the outcome of IVF-ET treatment.

Case-Control Studies ; Embryo Transfer ; Female ; Fertilization in Vitro ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; physiopathology ; Humans ; Male ; Pregnancy ; Pregnancy Outcome

OBJECTIVETo investigate the association of gonadotrophin (Gn) dose and ovarian response with the clinical outcome of in vitro fertilization and embryo transfer (IVF-ET).

METHODSPatients undergoing IVF-ET with Gn stimulation for no more than 15 days were enrolled in this study. The patients were divided into 3 groups, namely group A (390 cycles) with total Gn dose :3375 IU and retrieved oocytes:4, group B (64 cycles) with total Gn dose :3375 IU and retrieved oocytes < or =3, and group C (97 cycles) with total Gn dose< or =3300 IU and retrieved oocytes< or =3. The clinical characteristics and outcomes of these 3 groups were comparatively analyzed.

RESULTSThe clinical pregnancy rate and delivery rate were 38.8% and 32.5% in group A, 16.7% and 10.4% in group B, and 27.3% and 23.4% in group C, respectively. The follicle number, oocyte number, number of embryo transferred, peak serum E2 level, clinical pregnancy rate and delivery rate were significantly higher in group A than in groups B and C (P<0.05). Groups B and C had similar follicle number, oocyte number, and number of available embryos, but group C had significantly lower total Gn dose (P<0.05); the peak serum E2 level, clinical pregnancy rate and delivery rate were lower in group B than in group C, but the difference was not statistically significant (P>0.05).

CONCLUSIONSIn patients receiving a relatively low dose of Gn with smaller number of retrieved oocytes, Gn dose increment can improve the clinical pregnancy rate and delivery rate, suggesting a state of relatively poor ovarian response or mild ovarian reserve decrease; failure of increasing the number of oocytes retrieved with greater Gn dose suggests severely decreased ovarian responsiveness or ovarian reserve and also poor clinical prognosis.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Gonadotropins ; administration & dosage ; pharmacology ; Humans ; Infertility, Female ; physiopathology ; therapy ; Ovarian Follicle ; drug effects ; physiopathology ; Ovary ; drug effects ; physiopathology ; Ovulation Induction ; methods ; Pregnancy ; Pregnancy Outcome

OBJECTIVETo determine whether oxygen-glucose deprivation (OGD) induces C6 cell injury, and whether 5-lipoxygenase (5-LOX)/cysteinyl leukotriene (CysLT) pathway is involved in OGD-induced injury.

METHODSAfter OGD treatment and recovery for various durations, the viability of C6 cells was determined, and the effects of 5-LOX inhibitors and CysLT receptor antagonists were investigated. Intracellular distribution of 5-LOX protein was detected by immunocytochemistry, and the mRNA expressions of CysLT1 and CysLT2 receptors were detected by RT-PCR. The effect of leukotriene D(4) (LTD(4)) on C6 cells was also investigated.

RESULTOGD for 4-8 h followed by recovery for 24-72 h significantly induced C6 cell injury. Neither 5-LOX inhibitors nor CysLT receptor antagonists inhibited OGD-induced injury. OGD did not induce 5-LOX translocation into the nuclear membrane. C6 cells highly expressed CysLT(2) receptor, but the expression of CysLT1receptor was much weaker; the expression was not affected by OGD. In addition, LTD(4) did not affect C6 cells significantly.

CONCLUSIONOGD can induce C6 cell injury, but 5-LOX/CysLT pathway might be not involved in OGD-induced injury.

Animals ; Arachidonate 5-Lipoxygenase ; metabolism ; Cell Hypoxia ; physiology ; Glioma ; pathology ; Glucose ; metabolism ; Rats ; Receptors, Leukotriene ; metabolism ; Signal Transduction ; physiology ; Tumor Cells, Cultured