甲型血友病基因诊断及产前诊断的研究进展

甲型血友病是最常见的遗传性出血性疾病,受到了国内外研究者们的极大关注.由于此病尚无彻底根治方法,所以基因诊断及产前诊断显得尤为重要.基因诊断可分为直接基因诊断及间接连锁分析的方法;产前诊断的手段也在近年飞速发展.本文就直接基因诊断、间接连锁分析及产前诊断新技术这几方面的研究进展综述如下.     

小儿血友病93例临床分析

目的分析93例血友病患儿的诊断及治疗经验。方法对患儿的发病年龄、病例特点、实验室检查进行分析。结果血友病甲78例(84%),血友病乙15例(16%),其中重型45例(48%),中型33例(35%),轻型14例(15%),亚临床型1例(1%)。各种类型的出血为小儿血友病的主要表现,重型多见于婴幼儿,其中6例发生颅内出血(6%);关节病变及严重关节畸形多见于年长儿,共6例(6%)。初次出现出血症状的中位年龄为1.5(0～12.0)岁,但初次诊断的中位年龄为5.4(0.4～12.0)岁。延迟诊断的原因多种多样。结论延迟诊断、延误因子替代治疗是导致脏器出血和关节畸形的主要原因。     

Those with hemophilia or von Willebrand disease do not have increased perioperative complications following anterior cruciate ligament reconstruction: Results of a case-control large-database study

BackgroundThere is a lack of information on anterior cruciate ligament (ACL) reconstruction outcomes and complications for patients with congenital hypocoagulable conditions. The specific aim of this retrospective study was to report operative outcomes and complications for patients with congenital hypocoagulable disorders who underwent ACL reconstruction.MethodsWe performed a retrospective review of all patients who underwent an ACL reconstruction within Truven MarketScan Commercial Claims and Encounter Database from 2010 to 2014. Hemophilia A, hemophilia B and patients were identified. Patient demographics, cost of surgery, blood product use, concomitant injuries, repeat ACL injury, complications and various operative variables were collected. Statistical tests were conducted on SAS 9.4 2013.ResultsThirty-three hemophilia A, three hemophilia B, 63 von Willebrand factor patients, and 103,478 controls underwent ACL reconstruction. There is a statistically significant difference for hemarthrosis 1 year leading up to injury for hemophilia A compared with control (P = 0.0083). Total healthcare utilization 90 days after surgery was statistically significant for hemophilia A ($30,310 ± 52,745, P < 0.001) and von Willebrand factor ($20,355 ± 23,570, P < 0.001) compared with control ($14,564 ± 9512). Length of hospital stay, postoperative hemorrhage, concomitant injuries to the knee, additional ACL injury, infection rate, deep-vein thrombosis, and pulmonary embolism were not statistically significant. None of the hemophilia A or von Willebrand factor patients received blood products intraoperatively or postoperatively.ConclusionHemophilia A and von Willebrand factor patients had rates of postoperative complications and ACL re-injuries that were not statistically significant. Cost of healthcare utilization was identified as dramatically greater for hemophilia A and von Willebrand factor patients.     

连锁分析在血友病甲基因诊断中的应用

目的综述及讨论近几年以来连锁分析在血友病甲基因诊断中的应用的研究进展,为血友病甲连锁分析位点的选择提供有利的参考。方法以网络数据库资源为主,查询ScienceDirect,pubmed等数据库关于血友病甲连锁分析研究进展等方面的资料。结果共收集到多篇文献,选择其中16篇进行归纳总结与讨论。结论连锁分析现在在世界范围内都应用广泛,对血友病甲携带者的检出与产前诊断不失为一种简便、快捷实用的方法。但是依然有其局限性,比如,有些多态性标记并不能提供诊断信息,对于一些散发病例,连锁分析则不一定有效。对于基因外的多态性标记,细胞减数分裂期间基因重组会导致连锁分析错误。因此,有待发现更多有效的遗传多态性标记用于血友病甲的连锁分析。     

Synergistic inhibition of PARP‐1 and NF‐κB signaling downregulates immune response against recombinant AAV2 vectors during hepatic gene therapy

Host immune response remains a key obstacle to widespread application of adeno‐associated virus (AAV) based gene therapy. Thus, targeted inhibition of the signaling pathways that trigger such immune responses will be beneficial. Previous studies have reported that DNA damage response proteins such as poly(ADP‐ribose) polymerase‐1 (PARP‐1) negatively affect the integration of AAV in the host genome. However, the role of PARP‐1 in regulating AAV transduction and the immune response against these vectors has not been elucidated. In this study, we demonstrate that repression of PARP‐1 improves the transduction of single‐stranded AAV vectors both in vitro (～174%) and in vivo (two‐ to 3.4‐fold). Inhibition of PARP‐1, also significantly downregulated the expression of several proinflammatory and cytokine markers such as TLRs, ILs, NF‐κB subunit proteins associated with the host innate response against self‐complementary AAV2 vectors. The suppression of the inflammatory response targeted against these vectors was more effective upon combined inhibition of PARP‐1 and NF‐κB signaling. This strategy also effectively attenuated the AAV capsid‐specific cytotoxic T‐cell response, with minimal effect on vector transduction, as demonstrated in normal C57BL/6 and hemophilia B mice. These data suggest that targeting specific host cellular proteins could be useful to attenuate the immune barriers to AAV‐mediated gene therapy.     

联合运用St14(DXS52)位点VNTR和 FⅧ基因内的(CA)n重复多态性诊断甲型血友病

目的　提高甲型血友病 (hemophilia A,HA)家系成员基因诊断及产前基因诊断的准确性和可诊断率。方法　采用 St14 (DXS5 2 )位点的可变串联重复序列和 F 基因第 13内含子的 (CA) n重复多态性连锁分析对 HA家系进行间接基因诊断。结果　单用上述 2个多态位点中的 1个对 9个 HA家系进行连锁分析 ,可诊断率均为 6 6 .7% ,联合 2个多态位点 ,可诊断率则提高到 88.9% ,完成了 4个家系的产前基因诊断 ,并监测到 1例单用 St14位点的可变串联重复序列多态连锁分析可能发生的产前诊断的误诊。结论联合采用上述 2个多态位点可以对近 90 %的 HA家系作出快速、准确的基因诊断和产前基因诊断。     

血友病A患者FⅧ基因联合诊断及其DNA测序突变分析

目的探讨血友病A(HA)患者及携带者的基因诊断方法及诊断率。方法对20例HA患者及家系成员采用长距离PCR扩增(LD-PCR)技术、限制性内切酶酶切位点连锁分析(BclⅠPCR/RFLP)、可变串联重复序列多态性分析(St14 VNTR/PCR);并对FⅧ基因18号内含子的BclⅠ区段、19号内含子的HindⅢ相应区段进行DNA测序。结果检出22号内含子倒位携带者1例和倒位者6例;BclⅠPCR/RFLP酶切位点突变患者3例,携带者5例;St14 VNTR家系的连锁分析,发现携带者6例,患者5例;联合诊断家系总阳性率为90.0%;DNA测序两个区段突变位点分析,共发现5种突变。结论采用联合基因诊断方法,对FⅧ基因的3个位点进行联合基因诊断,可提高HA的诊断率及准确率;用PCR-DNA测序法可发现5种突变,并且这些突变位点在国内报道较少,22例患者突变检出率达50.0%。     

Development of samarium [P] phosphate colloid for radiosynoviorthesis applications: Preparation, biological and preliminary clinical studies experience

A new therapeutic radio colloid for radiosynoviorthesis (RS) applications is reported. The method of preparation involves the reaction of SmCl₃ carrier with carrier added [³²P]H₃PO₄ in the presence of gelatin. The pure colloid was recovered by dialysis purification leading to radiochemical yield of around 90%. The radiochemical purity of the pure colloid formulated in isotonic saline was over 98%, for the usage period of 14 days, as assessed by paper chromatography. Ninety percent of colloid particles were in the size of 1–10 μm as evident from the laser diffraction particle size analysis, ideally suitable for the intended end use. Animal studies revealed complete retention of the radio colloid in the rabbit knee joint. The results of clinical trials in humans are satisfactory and encouraging, satisfactory retention of the colloid in the knee joint and negligible leakage into the systemic circulation.