Objective To study the prenatal genetic diagnostic methods for hemophilia A fetus.Methods From 2002 to 2006,19 hemophilia A families were diagnosed either by long distance-polymerase chain reaction(LD-PCR)for factor Ⅷ intron 22 inversion or by the DNA polymorphism genetic linkage analysis of factorⅧin the Beijing Chaoyang Hospital.Results (1)Totally 19 women,with 22pregnancies received the prenatal diagnosis of fetal hemophilia A.The average week at diagnosis was 23(17-34)weeks.All the direct fetal blood sampling(DFBS)were successful.There was no fetal-loss caused by the procedures.(2)Of the 19 hemophilia A families,14 appeared to be factorⅧintron 22inversion,in which 16 prenatal diagnoses were done,10 fetuses were diagnosed as genetical hemophilia A patients,and 6 fetuses were normal.(3)Using combined polymorphism genetic linkage analysis 6 prenatal diagnoses were done,including one woman's two pregnancies,in which both her fetuses were diagnosed as genetical hemophilia A patients.(4)Factor Ⅷ levels of 16 fetuses were measured,and 6 fetuses were unmeasured either because the pregnancy weeks were lower than 20 weeks or the parents refused.Factor Ⅷlevel ranged from 0 to 198%.There were 11 fetuses whose factor Ⅷ levels were lower than 10%.Ten of them were diagnosed to be genetical hemophilia A patients,and in only one boy the factorⅧlevel was 2%,but the genetic diagnosis was normal and for one year's follow up he was doing normal.Conclusion LDPCR combined with polymorphism genetic linkage analysis enables a quick and correct detection ofhemophilia A carrier.For a carrier pregnancy.prenatal diagnosis could be done for the male fetus.Factor Ⅷ deficiency of the fetus could help make the diagnosis but the final diagnosis should be based on genetic evidence.

Objective To explore the aging mechanisms of human adult adipose-derived stem cells (hADSCs) in vitro.Methods The hADSCs came from the production of normal patients with liposuction.The hADSCs were cultured in the 37℃ incubator and passed until the cells were senescent.The senescence-associated β-gal staining (SA-β-gal) and histone protein γH2AX staining were carried out in both young and old hADSCs.The quantitative real-time PCR (qRT-PCR) was conducted to check the difference of the transcription function of the young and old hADSCs.Results The cultured hADSCs would be the senescent cells when the PD number was more than 35.The aging cells became big and had large flat nuclei and much more lososomes in the cytoplasm.The SA-β-gal staining and γH2AX staining were mostly positive in the aging hADSCs (PD>35) compared with the self-renewing hADSCs (PD<20).The qRT-PCR results showed that the transcription levels of Nanog and Oct4 gene were very low in aging hADSCs compared with the self-renewing ones.Conclusions The hADSCs will be senescent (SEN) when cultured and passed in vitro for long time and the aging process is similar to that of stem cells in vivo.The aging model of hADSCs can be established by this method and will be very useful in the anti-aging drug, regeneration medicine, tissue engineering and stem cell therapy.

Objective To investigate the mechanism of sleeve gastrectomy combined with modified jejunoileal bypass (SG/MJIB) in improving the blood glucose of Goto-kakizaki (GK) rats.Methods According to the random meter method,the 28 GK rats were divided into 4 groups:sleeve gastrectomy with modified jejunoileal bypass (SG/MJIB),sleeve gastrectomy (SG),modified jejunoileal bypass (MJIB) and sham operation (SHAM).The changes of weight,food intake,fasting blood glucose,oral glucose tolerance test (OGTT),insulin tolerance test (ITT),plasma insulin and gastrointestinal hormonal (ghrelin and GLP-1) were monitored before and 16 weeks after surgery respectively.Results After surgery,the fasting blood glucose level in SG/MJIB group was obviously lower than that in SG,MJIB and SHAM groups (P＜0.05).From the 2nd week after surgery,OGTT of the SG/MJIB group was obviously improved compared to SG,MJIB,and SHAM groups (P＜0.02).In our post-op erative study,the insulin levels in SG/MJIB group were lower than those in SG,and SHAM groups (P＜0.05).Compared to MJIB and SHAM groups,ghrelin levels of SG/MJIB group were significantly decreased (P＜0.001),while GLP-1 levels of SG/MJIB group were higher than those of SG and SHAM groups (P＜0.01).Conclusions SG/ MJIB can improve insulin sensitivity and insulin secretion,and this effect is independent of the body weight and food intake.This study further validates that the gastrointestinal hormones play an important role in the pathogenesis and treatment mechanisms of type 2 diabetes mellitus.

We designed two pairs of primers and their corresponding TaqMan probes according to gH, gE gene of PRV. By optimizing the probe's concentration, Mg2+ concentration, primers concentration and sample DNA extraction, real-time fluorescent quantitative PCR (FQ-PCR) which can quickly identity field virus and vaccine virus of PRV was established. According to our results, the dynamic range of the FQ-PCR assay is between 10 x 10(1) copies/microL and 10 x l0(8) copies/microL, and the detection limit of FQ-PCR is 1.0 x 10(1) copies/microL, which is 100 fold higher than that of conventional PCR. We detected 60 doubtful tissue samples using the FQ-PCR assay, serum neutralization and conventional PCR. In conclusion, the FQ-PCR method is rapid, sensitive, specific and accurate, and can be used to detect field strains of PRV rapidly. The closed-tube format of the assay minimized the risk of contamination of subsequent reaction and the assay can be performed in 2 h or less. Development of real-time quantitative PCR provides the basis for the early and rapid detection and analyzing quantitatively the infectious degree of PRV.
Animals ; Fluorescent Dyes ; Herpesvirus 1, Suid ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Pseudorabies ; diagnosis ; prevention & control ; virology ; Pseudorabies Vaccines ; immunology ; isolation & purification ; Swine

Objective To compare the in-hospital and follow-up clinical results of percutaneous coronary intervention(PCI)and coronary artery bypass grafting(CABG)in patients of left ventricular dysfunction with coronary artery disease.Methods 147 patients with left ventricular dysfunction were divided into PCI group(n=60)and CABG group(n=87).Clinical,angiographic and revascularization data were collected for analysis.Patients were by SPSS 13.0 software.P value of less than 0.05 was considered statistically significant.Results In-hospital MACCE rates and mortality ofthe two groups were comparable[(6.7%vs 9.2%,P＞0.05)and(1.7%vs 8.0%,P＞0.05)].Multivariate Logistic regression analysis indicated that in-hospital MACCE risk of the two groups were similar(OR≥3.03,95%CI 0.27～34.48,P＞0.05).22-month follow-up showed no signficance in MACCE rates (16.0%vs 13.8%,P＞0.05)and in repeated revaseularization rates(8.O%vs 1.7%,P＞0.05)between the two groups.Multivariate Cox regression analysis indicated that follow-up MACCE risk of the two groups were comparable (HR≥1.35,95%C/0.44～4.13,P＞0.05).Conclusion In coronary artery disease patients with left ventricular dysfunction,PCI and CABG have similar in-hospital and long-tem MACCE rates.Long-terra effect of PCI would be further increased with the wide use of drug-eluting stents.