Objective To explore the DSA mainfestations of MoyaMoya disease.Methods 19 patients, underwent CT before DSA, showed intracranial hemorrhage. All patients were then examined by angiography via femoral artery approach. Results All cases were diagnosed as MoyaMoya disease through DSA. The findings of DSA showed characteristic manifestations as the following: 1. Stenosis or occlusion of the invoved arteries. 2. Smoke like capillary vascular network spreading from supraseller cistern to cerebral base. 3. Development of collateral circulation. Conclusions DSA is the main method for the diagnosis of MoyaMoya disease, CT can only localize the site of cerebral hemorrhage.

Objective To investigate the correlation between KLF14 gene rs4731702 locus polymorphism and gestational diabe-tes mellitus (GDM) and relation between its different genotypes with BMI and insulin resistance.Methods This study adopted the case-control method.One hundred pregnant women of GDM (GDM group) and one hundred healthy pregnant women (normal con-trol group) were randomly selected as the research subjects and performed the physical examination ,biochemical indicators detec-tion.HOMA-IR and HOMA-β were calculated.The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted.The genotyping of KLF14 gene rs4731702 locus in the two groups was performed.The genotypes and allele gene frequency were compared between the two groups and the GDM association analysis was conducted.Results The C alleles fre-quency and CC genotype frequency of KLF14 gene rs4731702 locus in the GDM group was significantly higher than that in the con-trol group ,the difference was statistically significant (P< 0.05).The patients with genotype CC in the GDM group had higher BMI ,FPG ,TG ,HbA1c and HOMA-IR as compared with those carrying genotype CT and TT ,the difference was statistically signif-icant(P<0.05).Also they had lower FINS ,HDL and HOMA-βas compared with carriers of genotype CT and TT ,the difference was statistically significant(P<0.05).The multivariate analysis showed that the genotype CC of KLF14 gene rs4731702 locus was closely related with GDM (P=0.005 ,RR=25.128).Conclusion The genotype CC of KLF14 gene rs4731702 locus plays a role in the pathogenesis process of GDM ,may be susceptibility genes for GDM ,which is also related to the abnormal lipid metabolism ,islet dysfunction and obesity.The polymorphism study of KLF14 gene rs4731702 locus helps to reveal the relation between lipid metabo-lism abnormality and insulin resistance with GDM onset.

Objective To evaluated the clinical outcomes of intravesical instillation resiniferatoxin(RTX) for the treatment of patients with idiopathic overactive bladder(IOAB).Methods 26 cases with IOAB were randomly divided into test(A,14 cases) and control(B,12 cases) groups.The patients in group A were treated by intravesical instillation with 40ml of 0.5% idocain retained within the bladder for 3 minutes at first and with 100ml of 100nmol/L RTX retained within the bladder for 30 minutes late.The patients in group B were treated by the same method,however,the furacilin solution(placebo) at a dilution 1∶5000 was used instead of 100nmol/L RTX.The efficacy(daily voiding frequency,urgent uresis,FDV,MCBC,Qmax) of the 2 groups were evaluated before,1 month and 3 months after treatment.Results The pre-treatment comparion with prost-treatmen at 1 month and 3 months of A group was significant(P0.05).14 patients(54%) had slightly stimulating symptom in the urethra or bladder,and were otherwise generally well tolerated.Conclusion Single administration of RTX is safe and effective in patients with IOAB.

ObjectiveTo investigate the effects of intravenous (iv) remifentanil infusion on myocardial oxidative stress in rats.MethodsOne hundred and eighty male SD rats weighing 250-300 g were randomly divided into 15 groups (n =12 each):group control (group C); group ischemic preconditioning (group IPC); group remifentanil preconditioning ( group RPC ) ; while ia iv remifentanil infusion groups,iv remifentanil was infused at 4 different rates ( 1,5,10,20μg· kg- 1 · min- 1 ) and each rate was maintained for 15,60 and 120 min respectively.Myocardial ischemia was induced by occlusion of left coronary artery anterior descending branch for 30 min followed by 120 min reperfusion in 6 rats in each group.In group IPC myocardial ischemia was preceded by 3 cycles of 5min ischemia-5min reperfusion;whilein group RPC3cycles of 5min remifentanil infusion at 5 μg· kg-1 · min-1 were applied at 5 min interval before ischemia.Six rats in which I/R was produced were sacrificed in each group,myocardial infarct size (IS) and the area at risk (AAR) were measured and IS/AAR was calculated.The left 6 rats in each group were sacrificed at the corresponding time point (the end of each treatment)and superoxide radical expression and MDA and nitrotyrosine contents in myocardium were determined.Results IS/AAR was significantly decreased in groups IPC,RPC,1 μg·kg-1 ·min-1 × 120 min,5 μg·kg-1 ·min-1 × 60or 120 min and 10 μg· kg- 1 · min- 1 × 60 min as compared with group C.Compared with group C,the myocardial superoxide radical expression was significantly up-regulated in groups 1 μg· kg-1· min-1 × 120 min, 5 μg·kg-1 ·min-1 ×60 or 120 min,10μg·kg-1 ·min-1 ×60 or 120 min and 20 μg·kg-1 ·min-1 × 15,60 or 120min,and myocardial MDA and nitrotyrosine contents were significantly increased in group 20 μg· kg-1 · min-1 ×15,60 or 120 min.ConclusionLonger duration of high rate remifentanil infusion can induce myocardial oxidative stress in rats.

BACKGROUND: There are differences in physical and biological activity between the antibody from mammals and egg yolk antibody (IgY) from chicken. IgY is acid- and heat-resistant, and can prevent and cure the infectious diseases in animals and human being, which is also benefit to develop routine diagnostic immunoassays. Conventional ELISA assay for IgY takes much more time than dot-immunobinding assay.OBJECTIVE: To detect the IgY stability byusing dot-immunobinding assay.DESIGN: Open trail.SETTING: Department of Transfusion, Kunming General Hospital of Chinese PLA.MATERTALS: The experiment was completed in the Kunming General Hospital of Chengdu Military Area Command of Chinese PLA from January to June 2006. Two White Leghorn hens (30 weeks old) were selected. HLA-A*0201 α chain served as the antigen. The total protein concentration of the purified antigen was 0.04 g/L with the molecular mass of 32 000(self-prepared); nitrocellulose filter (NC, import and divided); nonfat dry milk (Anyi Corp. No. 20051220); DAB (Boshide Corp.);caprylic acid (made by Shanghai Xinghuo Chemical Factory); ammonium sulfate (Shantou Guanghua Chemical product).METHODS: ①HLA-A*0201 α chain with the total protein concentration of 0.04 g/L was purified with egg yolk antibody,and identified by SDS-PAGE. ②1 μL antigen was spotted into the center of NC membrane and dried in the incubator at 37 ℃. Then the NC membrane was blocked in 1 mL PBST and put in the incubator at 37℃ with shaking in 90 r per minute for 15 minutes. Then the liquid was exchanged with 1 mL PBST and added the primary antibody at a final concentration of 10 mg/L. After 30 minutes shaking in the incubator at 37 ℃, the NC membrane was washed in PBST for three times. The second antibody, mouse anti-chicken IgY conjugated to horseradish peroxidase (HRP) was added and after 30 minutes incubation, the NC membrane was washed three times in PBST. Binding was revealed by incubation with a DAB reagent. A positive reaction was represented by adeep brown spot,Irdlcating that IgY had better activity; if the spot became lighter IgY lost part activity, and when the spot disappeared, the IgY lost a the activty.According to intensity (gray degree)of the dot compared tothe standard, the remained percent of activity of the IgY was calculated. ③IgY was adjusted to three different protein concentrations with PBS: 1, 0.1, 0.01 g/L and stayed at room temperature for four months. 10 μg lgY was taken out from each concentration sample every month to detect the activity by dot-immunobinding assay. ④IgY was put into seven EP tubes with 100 μL per tube and numbered 1-7. Number 1 to 3 was adjusted pH to 5, 3 and 2, respectively with 1 mol/L HCI; Number 4 to 6 was to 9, 11 and 12, respectively with 1 mol/L NaOH. The pH of number 7 was neutral without adding acid or base. The samples were stayed in incubator at 37 ℃ for 3 hours. 10 μg IgY from each tube was taken every hour to detect the stability at different pH by dot-immunobinding assay. ⑤IgY was added to six EP tubes (10 μL per tube) and numbered 1-6. Number 1-6 was put into waterbath at 30, 40, 50, 60, 70 and 80 ℃ for 15 minutes. After cooled in refrigerator at 4 ℃, 10 mg samples from each tube and standard sample (untreated sample) taken to check the thermal stability by dot-immunobinding assay.MAIN OUTCOME MEASURES: ①SDS-PAGE of IgY. ②IgY stability at room temperature. ③IgY stability at different pH. ④ Detection of IgY thermal stability.RESULTS: ①Purified IgY after SDS-PAGE had two major binds, the molecular mass of the heavy chain was 66.000,and the light chain was 25 000. ②1, 0.1, 0.01 g/L IgY still had partial activity after staying at room temperature for four months. ③When pH ranged from 5 to 9, IgY still had partial activity after staying in 37 ℃ for 3 hours. If pH was lower than 5 or higher than 9, it lost the whole activity in above condition. ④Purified IgY was added to six EP tubes, the number 1-4 still had partial activity, but number 5 and 6 showed some white precipitate, which was caused by protein denaturation at higher temperature.CONCLUSION: IgY stability is higher than others. The dot-immunobinding assay described a rapid and simple method for the demonstration and characterization of functional activity of egg yolk antibody. With only small volume antigen and antibody, and specific dot, the dot-immunobinding assay method could process many samples at the same time.

Acute Disease ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Myocarditis ; diagnosis ; diagnostic imaging ; Ultrasonography ; methods ; Virus Diseases ; diagnosis ; diagnostic imaging

Objective To investigate the value of Doppler ultrasound in the eady diagnosis,monitoring and assessing of renal damage in neonatal asphyxia.Methods A total of 60 cases of neonates within 24 h were divided into severe asphyxia group (Apgar score 0-3),mild asphyxia group (Apgar score 4-7) and healthy control group (Apgar score 8-10) according to Apgar score at 1 min after born.Then the peak systolic velocity (PSV),end diastolic velocity (EDV) and resistance index (RI) of renal artery were obtained by Doppler ultrasound within 24 h,on day 3,day 7,and day 10.The level of serum cystain C (sCysC) was also recorded accordingly.Results Within 24 h,compared with healthy control group,the PSV and EDV in severe asphyxia group and mild asphyxia group decreased (all P＜0.05),while RI increased (all P＜0.05).The PSV in mild asphyxia group returned to normal in 3 days,EDV and RI returned to normal in 10 days,there were no statistically significant difference compared with healthy control group (all P＞0.05).The PSV,EDV and RI in severe asphyxia group were still significantly differences compared with healthy control group on day 10 (all P＜0.05).Within 24 h and on day 3,sCysC in the mild asphyxia group increased obviously compared with healthy control group (both P＜0.05).On day 7 and day 10,the differences of sCysC was not statistically significant between mild asphyxia group and healthy control group (both P＞0.05).Compared with healthy control group,the sCysC in severe asphyxia group increased significantly (all P＜0.05) on every time point.PSV and EDV were negatively correlated with sCysC,RI was positively correlated with sCysC.Conclusion Changes in renal function can be reflected soon by index of renal blood flow PSV,EDV and RI.

Objective To investigate the diagnostic value of the recombinant surface antigen 1 (rSAG1) in immunodiagnosis of toxoplasmosis. Methods Isopropyl β-D- 1 -thio-galaetopyranoside (IPTG) was used to induce the expression of recombinant plasmid pET28a-SAG1 of Escherich coli(pET28a-SAG1/BL21 ). The expression products (rSAG1) of pET28a-SAG1/BL21 were identified by Western blotting. The serum of mice infected with Toxoplasma gondii tachyzoites, normal mouse serum and the serum from 10 toxoplasma gondii patients were used as primary anti-Toxoplasma gondii antibodies, and the rSAG1 gene products were identified by Western blotting, by which the diagnostic value of rSAG1 in Toxoplasmosis was compared. Results After induction and purification, rSAG1 protein was obtained and its relative molecular mass was 38.5 × 103. The fusion protein could be recognized by the serum of mouse infected with Toxoplasma gondii tachyzoites, rSAG1 of expression products of surface membrane antigen SAG1 gene from Toxoplasma Gondii could be detected in 4 cases from 10 patients by Westem blotting.Conclusion The rSAG1 has a potential value in the immunodiagnosis of Toxoplasmosis.

Background Hypoxia is a crucial factor of neovascularization.Many researches found that stromal cell-derived factor-1 (SDF-1) and integrin-linked kinase (ILK) play an important role in the neovascular disease.However,effect of SDF-1 and ILK in eye neovascular disease is below understood.Objective The aim of this study was to investigate the effect of hypoxia on the expressions of SDF-1 and ILK in cultured retinal pigment epithelium(RPE) cells in vitro.Methods RPE tissue was isolated from 4-week-old C57BL/6 mouse and was digested and cultured in DMEM/F12 with 10％ fetal bovine serum (FBS).The cells with 80％ confluence were collected and passaged.The third generation of cells were identified with cytokeratin 18 (CK18) antibody by immunochemistry.The cells were inoculated at the density of 5×104 cells/ml to free-serum DMEM/F12 for 24 hours and then were cultured in regular medium in the normoxic control group.RPE cells were cultured for 1 hour and 3,6,12,24,48,72 hours with 200 μmol/L CoCl2 in the hypoxia group.Reverse transcription-PCR(RT-PCR) was used to evaluate the expressing change of SDF-1 mRNA and ILK mRNA in RPE cells,and Western blot was used to assay the expressing change of SDF-1 protein and ILK protein in RPE cells in different time points.The detected outcomes were represented as the ratio of target gene A value/β-actin A value.Results Cultured cells showed the polygon in shape with the black pigment granules in cytoplasm.Over 90％ cells were positive response for CK18.Expressions of the SDF-1 mRNA and ILK mRNA were increased in different time points after CoCl2 co-cultured(SDF-1 mRNA:F=281.875,P=0.000 ;ILK mRNA: F=187.566,P=0.000),with the highest expressing value in hypoxia at 12 hours.No significant change in the expression of SDF-1 mRNA and protein was found 1 hour after CoCl2 co-cultured,but expressions of SDF-1 mRNA and ILK mRNA were significantly higher in 3,6,12,24,48 and 72 hours than the normoxic control group(P＜0.01).The expressions of SDF-1 protein and ILK protein were gradually ascended with the time increase of CoCl2 co-culture,showing a significant difference among different time points(SDF-1: F=44.719,P =0.000 ; ILK: F =144.481,P =0.000),and the up-regulation of SDF-1 protein and ILK protein expression was seen mainly in 3,6,12,24,48 and 72 hours after CoCl2 co-cultured in comparison with the normoxic control group (P＜0.01).Conclusions SDF-1 and ILK are involved in the hypoxic response of RPE cells and may play a potential role in ischemic/hypoxic retinopathy.

Objective To understand the erythromycin resistance rate and the erythromycin resistant gene spectrum in Streptococcus pyogenes strains isolated in Shanghai.Methods The outpatient children who were diagnosed with scarlatinal in the Children's Hospital of Fudan University from November 2004 to June 2006 were enrolled and 100 strains of Streptococcus pyogenes were isolated by pharyngeal swab culture.The distributions ofermA,ermB,mefA genes were determined by polymerase chain reaction(PCR)amplification.The relationship between ermA,ermB,mefA genes and erythromycin resistance were also analyzed.Results The erythromycin and clindamycin resistance rates of Streptococcus pyogenes in Shanghai were 98%and 95%,respectively;the concordance rate of these two drugs was 97%.Among 100 strains of Streptococcus pyogenes,94 strains(94%)contained ermB gene,with 100%of erythromycinresistance rate.Sixteen(16%)contained mefA gene,with 100% of erythromycin resistance rate.ermA was not detected inall the 100 strains.The ermB and mefA genes were not found in 5 strains,among which,2 were susceptible to erythromycin and 3wereresistant to erythromycin.Only 1%of isolates was mefA genesingle positive.Conclusions There is a high erythromycin resistance rate of Streptococcus pyogenes strains isolated inShanghai,and the cross resistance to clindamycin is high.TheermB gene is important erythromycin resistancedeterminants of Streptococcus pyogenes in Shanghai.