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 systematically review the efficacy and safety of acid-suppressive therapy including proton pump inhibitors (PPI) and histamine 2 receptor antagonists (H2RA) and compare with those of placebo or blank control in the postoperative patients with stress ulcer bleeding (SUB) to provide evidence-based reference for clinical practice.Methods:The Cochrane library,Medline,Embase,CBM,CNKI,VIP,Wan Fang Data,clinicaltrials.gov,ISRCTN Register and WHO ICTRP were searched.Only randomized controlled trials (RCTs) of acid-suppressive therapy compared with placebo or blank control for postoperative stress ulcer bleeding were selected with bleeding rate,mortality,adverse drug reactions,gastric juice pH and length of stay as the indices.After the quality evaluation and data extraction,Meta-analysis was performed by using Stata12.0 statistics software.The results were expressed as relative risk(RR) and its corresponding 95% confidence interval(CI).Funnel plot and Eggers test were used to determine the publication bias;and then Grade approach was applied to assess the confidence in the evidence for each outcome.Results:Totally 15 trials enrolling 971 patients were selected,and acid-suppressive therapy was more effective than placebo or blank control in reducing the risk of stress ulcer bleeding,overt upper gastrointestinal bleeding and clinical important bleeding(RR 0.29,95% CI:0.19-0.45;RR 0.25,95%CI:0.10-0.64;RR 0.36,95%CI:0.17-0.77)(moderate),however,there was no statistical significance in the incidence of occult bleeding,mortality and adverse drug reactions (RR 0.79,95%CI:0.41-1.50;RR 0.49,95%CI:0.17-1.38;RR 0.78,95%CI:0.33-1.85,very low confidence).The subgroup analysis of drug classification,operation types and administration juncture showed that the incidence of SUB using PPI (RR=0.27) was lower than that using H2RA (RR=0.33);that of heart surgery (RR=0.20) was lower than that of general surgery (RR=0.31) and neurosurgery(RR=0.37);that of postoperative administration (RR=0.26) was lower than that of preoperative administration (RR=0.23).Conclusion:Acid-suppressive drugs seem to be more effective than placebo or blank control in reducing the risk of bleeding without significant increase of the risk of mortality or adverse drug reactions.The robustness of the conclusion is limited because of the low quality of the trial methodology,incomplete outcome indicators and lack of safety indices for pneumonia and clostridium diffcile-associated infection.Trials with high-quality and larger sample size are still needed to verify its clinical effects.

Multidrug resistance proteins (MRP2, ABCC2) may play a role in drug resistance in epilepsy by limiting gastrointestinal absorption and brain access of antiepileptic drugs (AEDs). We sought to investigate the effects of ABCC2 polymorphisms on plasma carbamazepine (CBZ) concentrations and pharmacoresistance in Chinese patients with epilepsy. ABCC2 rs717620, rs2273697, rs3740066 polymorphisms were genotyped by polymerase chain reaction amplification followed by restriction fragment length polymorphism analysis or direct automated DNA sequencing in 80 patients treated with CBZ monotherapy. There were no differences in CBZ maintenance doses or adjusted plasma CBZ concentrations among the ABCC2 rs717620, rs2273697 and rs3740066 genotypic groups. No associations between all the studied genotypes and haplotypes involving the three SNPs of ABCC2 and CBZ resistance were observed in this patient cohort. These results suggest that ABCC2 polymorphisms may not contribute to interindividual variabilities in CBZ daily maintenance doses, plasma concentrations, and treatment efficacy.
Epilepsy

Objective To study the polymorphisms of short tandem repeat (STR) loci in intron 1, 24, 13 and 22 (STR 1,24, 13, 22) of factor Ⅷ (FⅧ) gene in Chinese population, and establish single tube multiple fluorescent PCR method for rapid diagnosis of haemophilina A(HA). Methods Four STRs from genomie DNA of 220 females without blood relationship were amplified in a single tube using quadri-fluorescence PCR. Capillary electrophoresis was analyzed in ABI PRISM 310 Genetic Analyzer. DNA sequencing was used to assay the number of dinueleotide repeats. Gene diagnosis were performed in 96 HA families. Results It was observed that 7, 9, 10 and 7 different alleles were found in STR1, 24, 13 and 22, respectively. The PIC (polymorphism information contents) were 0. 3789, 0. 4055, 0. 5239 and 0. 4713 in STR1,24, 13 and 22, respectively, and the HR (heterozygesity rate) were 34. 55% (76/220), 38. 18% (84/220), 49. 55% (109/220) and 43.64% (96/220). In 96 HA families, the diagnosis rate of STR1, 24, 13 and 22 were 38. 54% (37/96), 38. 54% (37/96), 54. 17% (52/96), 42. 71% (41/96), respectively. Whereas it achieved 79. 17% (76/96) when combining the four STR markers. Conciusion The single tube multiple fluorescent PCR of four STR loci is an effective, simple, quick method for linkage analysis and gene diagnosis of haemophilia A.

Objective To explore the significance in judging the different clinical stages of relapsing polychondritis (RP) patients through examining the changes of aggrecanase and metabolic fragments of aggrecan.MethodsIn comparison with the control group (20 cases),40 patients were divided into the stable stage group (22 cases) and the active stage group (18 cases).The aggrecanase-generated neoeptitopes in cartilage matrix were analysed by immunohistochemistry and Western blot(WB) respectively.The mRNA and protein levels of aggrecanase-1,2 expressed in cartilage cells were measured by real-time reverse transcriptional polymerase chain reaction(RT-PCR) and WB respectively.The difference of these results among these three groups was analyzed accordingly.ResultsThe expression of aggrecanase-1,2 in mRNA level was measured by real-time RT-PCR.The values of aggrecanase-1,2 mRNA 2 -ΔΔC1 were 1.00 ± 0.26 and 1.00 ± 0.27 in control group,1.47 ± 0.11 and 1.57 ± 0.13 in stable stage group,2.09 ±0.12 and 2.09 ± 0.19 in active stage group respectively.By one-way ANOVA analysis,the difference between every two groups was statistically significant (F was 299.113 and 459.013,P ＜ 0.01 ).In comparison with control group,aggrecanase-1,2 increased significantly in both stable and active stage group (P ＜ 0.01 ) and aggrecanase-1,2 increased more significantly in active stage group than in stable stage group (P ＜ 0.01 ).The results from WB analysis indicated that aggrecanase-1,2 could not be detected in control group,and they were detectable in stable stage group and increased in active stage group at the relative molecular of 68 000 Da or 73 000 Da respectively.The aggrecanase-generated neoeptitopes were analyzed by WB as well.The results indicated that NITEGE and ARGSV could be detected in stable stage group and increased in active stage group at the relative molecular of 70 000 Da or 48 000 Da respectively,but there were no signals in control group.Similar with the previous WB results,no signals of NITEGE or ARGSV eptitopes were detected in normal cartilage matrix ( no red staining) by use of immunohistochemical staining.However,in stable stage group and active stage group,these eptitopes were apparently detected (obviously red staining).ConclusionWith the progression of the RP,the activity of the aggrecanase is enhanced,and the degradation of the aggrecan is increased,associated with the severity of the disease.

Bupleurum ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective To investigate the effect of allicin on the development of atherosclerosis in apoE-/-mice and explore its underlying mechanism from the perspective of protein S-nitrosylation.Methods Thirty male apoE-/- mice were randomly divided into 3 groups:control group (saline,ig),low-dose group (allicin,9 mg/kg·d, ig)and high-dose group (allicin,18 mg/kg·d,ig).They were fed with high cholesterol diet for 12 weeks.The levels of plasma lipids,oxidized-LDL (ox-LDL),malondialdehyde,tumor necrosis factor-alpha and nitric oxide (NO)were measured.The atherosclerotic lesions in aortic root were evaluated after hematoxylin and eosin staining and elastica van Gieson and immunohistochemical staining,respectively.Furthermore,in vitro experiments were performed using human umbilical vein endothelial cells (HUVECs).The HUVECs were treated with allicin (10μmol/L or 20 μmol/L)for 24 hours in the presence of ox-LDL (50 μg/mL).The level of NO in supernatant was measured by a nitrate/nitrite assay. The protein S-nitrosylation of the HUVECs was detected through immunofluorescence.Results The histological analysis revealed that allicin treatment not only significantly decreased the areas of the atherosclerotic lesion (all P <0.05)but also suppressed the macrophage accumulation and smooth muscle cell proliferation in the lesion.There was no significant difference in the levels of plasma lipids between control and treated groups.However,allicin exerted obvious anti-oxidative and anti-inflammatory effects. Interestingly,the allicin treatment led to marked increase of the plasma NO level (P <0.05)and aortic protein S-nitrosylation.The experiments in vitro further proved that the allicin up-regulated the levels of NO and protein S-nitrosylation in HUVECs treated with ox-LDL (P < 0.01 ).Conclusion Allicin can inhibit the development of atherosclerosis.The mechanism is associated with the up-regulation of protein S-nitrosylation in endothelial cells, which plays an important role in anti-oxidization and anti-inflammation.

Objective This study is to summarize the experience of microsurgical reconstruction for donor liver anatomical variations of hepatic arteries in orthotopic liver transplantation. Methods During the bench surgery, the anatomy of donors′ hepatic arteries was carefully examined and microsurgical techniques were used for the anomalous arteries. The graft arterial flow was checked by Doppler ultrasound daily in the first week in postoperative period and periodically thereafter. Results The arterial anatomy was anomalousin 20 out of 141 (14%) donor livers. Nine cases (6.3%) needed arterial reconstruction. In these cases, 7(4.9%) aberrant right hepatic arteries originating from superior mesenteric artery were anastomosed to gastro-duodenal arteries and another two aberrant hepatic left or right arteries were anastomosed to the stump of the donor splenic arteries. Conclusions The variations of hepatic arteries in donors are common. To obtain the ideal arterial supply of liver graft, both careful checking on the origin of donor's artery and appropriate plastic performance with refined microsurgical techniques are necessary.

Objective To investigate the Mycobacterium tuberculosis infections in 2014 among Beijing army recruits, and evaluate a new method for screening latent tuberculosis infections.Methods A total of 194 army recruits were subjected to chest X-ray examination purified protein derivative(PPD) skin test, antibody detection, and interferon gamma release assay(IGRA) by ELISA combined with recombinant protein CFP10-ESAT6 and latent infection protein Rv2628.Results The positive rates of PPD skin test and antibody test were 49.7% and 15.5%, respectively.The latent infection rate of IGRA test was 22.2% in 194 cases after CFP10-ESAT6 stimulation.After stimulation of latent tuberculosis infection(LTBI) with Rv2628, IFN-γ level was significantly higher than that in healthy control group (P<0.05).The weak positive group of TST (5 mm≤diameter<15 mm) had a significantly higher level of IFN-γ than the strong positive group(diameter≥15 mm)(P<0.05),but after stimulation with CFP10-ESAT6,IFN-γ levels were not significantly different between the two groups(P>0.05).There was no significant difference between antibody negative and positive groups after stimulation by CFP10-ESAT6 and Rv2628 (P>0.05).The area under the ROC curve of Rv2628 diagnosis of tuberculosis infection was 0.84.When Youden index was 0.621,the specificity was 94.7% and sensitivity was 67.4%.ConclusionCombined detection of antigens Rv2628 and CFP10-ESAT6 specific IFN-γ values can be potentially used for differential diagnosis of active or latent tuberculosis infections.

Objective To compare the difference in radiotherapy dose caused by different ways of adding bolus.Methods A total of 20 patients who needed to receive postmastectomy chest wall irradiation from October to December on 2014 were selected.Each patient underwent two CT scans;CT-1 was to perform CT scan directly without bolus, and CT-2 was to perform CT scan after adding bolus to the body surface.An equivalent bolus was added for CT-1 in the radiotherapy planning system, and Plan-1, which met the clinical requirements, was performed.Then Plan-1 was put on CT-2 through image fusion and plan verification to develop Plan-2, which was to develop plans with equivalent boluses at other times and perform radiotherapy with a bolus added to the surface of the body.At last, CT-2 was used to perform radiotherapy Plan-3, which met the clinical requirements.The paired t-test was used for comparison of clinical data between any two plans with SPSS 19.0.Results The V20 of the whole lung, V20 of the diseased lung, V30 of the heart, and Dmax of the healthy breast showed no significant differences across the three plans (P=0.074-0.871).The V50 , V55 , conformity index, and homogeneity index of the planning target showed significant differences across the three plans, and the total number of monitor units showed a significant difference between Plan-1 and Plan-2(P=0.002-0.049).The dose distribution in the target volume and the number of monitor units in each radiation field also showed significant differences.Conclusions When the equivalent bolus is added to the body surface before CT scan, such a plan can accurately reflect the dose distribution of the planning target and the dose to organs at risk.