3例自身免疫性血友病样凝血因子缺陷症ⅩⅢ患者诊断流程的初步建立及文献回顾

目的 :对3例疑似自身免疫性血友病样凝血因子ⅩⅢ缺陷症(autoimmune haemorrhaphilia factorⅩⅢ,AHⅩⅢ)患者进行检测,明确其临床诊断、治疗及随访资料,建立完整的AHⅩⅢ实验诊断流程。方法:收集3例AHⅩⅢ患者的临床信息,通过氨释放法检测其凝血因子ⅩⅢ(factorⅩⅢ, FⅩⅢ)活性,酶联免疫吸附试验(enzyme linked immunosorbent assay, ELISA)法检测FⅩⅢ-A抗原(FⅩⅢ-A:Ag)、FⅩⅢ-B:Ag。抗FⅩⅢ抗体检测包括混合实验检测抗FⅩⅢ中和抗体的存在、Bethesda法测定抗FⅩⅢ抗体滴度、ELISA法检测抗FⅩⅢ-A亚基抗体等。结果:3例患者的尿素法FⅩⅢ定性试验均为阳性,FⅩⅢ活性5%,FⅩⅢ-A:Ag3%,FⅩⅢ-B:Ag分别为65.40%、116.28%、138.81%。3例患者混合实验均为阳性,Bethesda法测定抗FⅩⅢ中和抗体滴度分别为1.83 BU、1.67 BU、1.75 BU,免疫法检测结果提示均存在抗FⅩⅢ-A亚基抗体。结论:建立了完整的ⅩⅢ的诊断流程,并回顾了其诊断、治疗及预后相关文献。经诊断,本研究3例患者均为低滴度抗FⅩⅢ-A亚基抗体导致的AHⅩⅢ。     

获得性凝血因子Ⅷ抑制物对获得性血友病A的诊断意义:附1例病例分析

横县中医医院于2018年10月28日收治1例获得性血友病A(AHA)患者,该患者因面色苍白10余天,右侧大腿及膝关节肿胀,局部可见片状瘀斑,拟"贫血原因待查"收入院。经活化部分凝血活酶时间(APTT)测定、APTT纠正试验、凝血因子Ⅷ抑制物测定(Bethesda法)、凝血因子Ⅷ水平测定等发现,该患者APTT显著延长,APTT纠正试验不能被纠正,血小板计数(PLT)和肝功能正常;经进一步检测发现,患者凝血因子Ⅷ活性为1%,凝血因子Ⅷ抑制物为9.6 BU/mL(Bethesda法);抗核抗体(ANA)为核颗粒型(1:320),ANA谱抗SSA抗体阳性,抗SSB抗体阳性。故诊断为获得性凝血因子Ⅷ抑制物所致出血。当患者临床表现为不明原因贫血伴可见片状瘀斑时,应考虑是否伴有获得性因子Ⅷ抑制物存在的可能;APTT纠正试验对诊断是否存在循环抗凝抑制物意义重大。     

血清β-CTx及TRACP-5b与多发性骨髓瘤骨病病情严重程度及预后关系

目的 探讨血清Ⅰ型胶原羧基端肽β特殊序列(β-CTx)及抗酒石酸酸性磷酸酶5b同工酶(TRACP-5b)与多发性骨髓瘤骨病(MMBD)病情严重程度及预后的关系。方法 回顾分析2016年1月-2020年1月收治的MMBD患者142例。通过诊治指南和影像学检查结果将患者按病情严重程度分为1、2、3、4级组,采用酶联免疫吸附法(ELISA)检测4组血清β-CTx、TRACP-5b水平,比较4组血清指标变化情况,以Spearman分析血清β-CTx、TRACP-5b与MMBD严重程度的相关性。所有患者根据生存情况分为生存组与死亡组,采用Cox回归分析影响预后的相关因素。结果 血清β-CTx、TRACP-5b水平随4组病情严重程度等级的增加而升高(P<0.05)。Spearman分析显示,血清β-CTx、TRACP-5b水平均与MMBD病情严重程度呈正相关(r=0.753、0.627,P<0.05)。142例患者截至随访时间生存98例,死亡44例,死亡组国际分期体系(ISS) Ⅲ期占比、MMBD严重程度4级占比、病理性骨折骨病类型占比、血清β2-MG、β-CTx及TRACP-5b水平较生存组更高(P<0.05)。β-CTx/TRACP-5b高、低水平的患者生存曲线差异具有统计学意义(P<0.05)。Cox回归分析显示,血清β-CTx、TRACP-5b、β2-MG水平、ISS Ⅲ期、MMBD严重程度4级、病理性骨折骨病类型是影响MMBD患者预后的危险因素(P<0.05)。结论 血清β-CTx、TRACP-5b水平随MMBD病情严重程度的增加而升高,且二者是影响MMBD预后的危险因素。     

抗着丝点抗体在肝病患者中的检测及其意义

目的探讨抗着丝点抗体(ACA)的肝功异常患者中出现的频率、特点及其临床意义。方法对10000例肝功异常患者采用间接免疫荧光法检测抗核抗体(ANA)、抗线粒体抗体(AMA)、抗平滑肌抗体(SMA)和抗细胞骨架抗体等,并采用免疫印迹法检测抗着丝点抗体及其它抗可提取的核抗原(antextractablen2uclearantigens,ENA)抗体。结果 (1)10000例肝功异常患者中,间接免疫荧光法筛选结果显示抗核抗体阳性,着丝点型的患者为134例,占1.34%。其中滴度1∶100的占35.8%;滴度1∶320的占40.3%;滴度大于等于1∶1000的占23.9%。滴度大于等于1:320的患者再免疫印迹法检测,抗着丝点抗体均为阳性。部分患者伴有其他核型的抗核抗体以及抗线粒体抗体等自身抗体。(2)134例ACA阳性患者,诊断为硬化型肝病的有83例,包括PBC、PBC和AIH重叠症、肝炎肝硬化以及原发性肝癌硬化型。诊断为非硬化型肝病的有51例,包括病毒性乙型肝炎、自身免疫性肝炎、药物性肝炎和病毒性丙型肝炎等。抗着丝点抗体高滴度组患者的肝硬化发生率明显高于低滴度组。(3)分析高滴度组(滴度大于等于1∶320)和低滴度组(滴度1∶100)组的实验室检查结果发现,ACA高滴度组AST/ALT比值明显高于低滴度组,ALB和PA明显低于低滴度组,IgM的含量也高于低滴度组,两组间的差别有统计学意义。结论肝病患者中,抗着丝点抗体并不少见,高滴度的抗着丝点抗体更见于各种原因引起的肝硬化患者。     

“组团式”紧密型帮扶提升基层中医院护士中医护理服务能力的实践

目的 探索“组团式”紧密型帮扶提升基层中医院护士中医护理服务能力的方式及效果。方法 充分了解基层中医院的需求,成立中医护理帮扶专家团,并派驻中医护理骨干紧密型驻点帮扶。采取“双”式和“播种”式帮扶,强化人才培养;建立护理服务质控体系,引入科学管理理念;“手把手”指导+“传帮带”教学,促进学科发展;“师带徒”模式打造联动门诊,帮助基层医院提供高质量服务。结果 被帮扶医院中医护理服务能力显著提高,中医理论与操作评分较帮扶前明显提高（P<0.001）,其中中药涂药技术、蜡疗技术和经穴推拿技术3项操作进步最大;被帮扶医院护士对“组团式”紧密型帮扶总体满意度、教学计划安排满意度、授课内容满意度均达到99%以上;被帮扶医院成功开设中医特色护理门诊,培养了8名院级中医护理骨干人才。结论 对基层中医院实施“组团式”紧密型帮扶可有效提升被帮扶医院的中医护理服务能力,进而提高基层中医院中医护理服务质量。     

全血γ-干扰素释放试验与ELISA检测结核分枝杆菌抗体在痰菌阴性肺结核辅助诊断中的比较应用

目的：探讨γ-干扰素释放试验（interferon-gamma release assays, IGRA）与酶联免疫吸附试验（enzyme linked immunosorbent assay, ELISA）在痰菌阴性肺结核诊断中的应用价值。方法：收集93例痰菌阴性肺结核患者和40例健康体检者的肝素钠抗凝全血,采用IGRA试剂盒检测其血浆γ-干扰素含量,其血清标本应用ELISA试剂盒检测结核分枝杆菌抗体（Tuberculosis antibody, TBAb）,并将2种结果进行比较分析。将痰菌阴性肺结核患者分为老年组和中青年组进行比较,分析IGRA检测在不同年龄段间是否有差异。结果：IGRA诊断痰菌阴性肺结核,灵敏度为84.94%,特异度为90.00%,阳性预测值为95.18%,阴性预测值为72.00%;TBAb诊断痰菌阴性肺结核灵敏度为52.69%,特异度为85.00%,阳性预测值为67.20%,阴性预测值为40.96%。2种方法相比较,差异有统计学意义。不同年龄患者间,IGRA检测结果差异无统计学意义。结论：与ELISA相比,IGRA对诊断痰菌阴性肺结核有更高的灵敏度、特异度和阳性预测能力。     

盐水法接合柱凝集法在ABO疑难血型检测中的应用

目的解决柱凝集法检测ABO血型抗原减弱或抗体减低所产生的正反定型不一致。方法将疑难血型患者2%红细胞50μl与50μl抗-A、抗-B标准血清反应,或者将患者的血清(血浆)50μl与ABO试剂细胞A1、B各50μl反应,然后把反应液接合到Liss/Coombs(Anti-IgG+C3d)卡上进行测定;同时利用此法进行抗-A、抗-B标准血清最大稀释度的检测。结果盐水法接合柱凝集法检测17例抗原弱化或抗体减低的疑难血型,全部出现明显阳性反应并符合血型鉴定结果,以此法检测抗-A、抗-B标准血清最大稀释度达到1∶4096。结论盐水法接合柱凝集法测定抗原弱化或抗体减低的ABO血型是一种高效、灵敏、简单易行的方法。     

婴幼儿ABO血型IgM抗-A、抗-B产生及效价分布调查

目的了解<6月龄婴幼儿ABO血型IgM抗-A、抗-B产生顺序及效价范围。方法选取北京儿童医院<6月龄住院婴幼儿,采集不抗凝静脉血标本,应用单克隆抗-A、抗-B、A1细胞、B细胞及O细胞进行试管法血型检定,应用试管倍比稀释法进行IgM抗体效价滴定。结果 ABO血型正定型为A型婴幼儿标本781例,其中抗-B437例(55.95%);正定型为B型婴幼儿标本795例,其中抗-A 498例(62.64%);正定型为O型婴幼儿标本498例,其中抗-A 304例(61.04%),抗-B 276例(55.42%)。在已检出有抗体的婴幼儿中,76.32%～85.81%的婴幼儿IgM抗体效价仅在原管凝集,高效价者中抗-A例数多于抗-B。结论婴幼儿体内IgM抗-A的产生早于IgM抗-B,且前者效价略高于后者。     

血友病凝血因子抗体的诊断及影响因素研究

本研究探讨血友病凝血因子抗体(抑制物)的诊断、分型及影响因素.对500例血友病采用一期法检测FⅧ活性(FⅧ:C),FⅨ活性(FⅨ:C),用Bethesda法检测FⅧ抗体(FⅧ:Ab)、FⅨ抗体(FⅨ:Ab).结果表明,500例血友病中,HA 411例,FⅧ抗体阳性者151例(30.2%),滴度为3.50±2.84 Bu/ml;HB 79例,FⅨ抗体阳性者18例(3.6%),滴度为(2.92 ± 2.19)Bu/ml;自身获得性血友病10例(2.0%).抗体分型:高、中、低反应型依次为3例、47例、119例,以中低反应型抗体为主(98.22%).抗体产生的影响因素:169例抗体阳性者的92.90%(157/169例)年龄在30岁以下;89.35%(151/169例)为HA;81.66%(138/169例)为中、重型血友病;98.22%(166/169例)为中、低反应型抗体;同种特异性杭体阳性者158例均有输血史.结论:抗体以中、低反应型为主,患者年龄及输血史是影响的因素.本研究为血友病凝血因子抗体的诊断、分型及其产生的影响因素提供了依据,并说明血友病反复输注血液制品对抗体的产生有直接影响,对防治血友病出血及预防凝血因子活性降低有重要意义.     

微柱凝胶技术检测521例O型血孕妇IgG A(B)抗体效价

目的研究血型为O型的孕妇产前IgG A(B)抗体效价,以预防新生儿溶血病的发生。方法采用微柱凝胶法测定IgG抗-A(B)效价。结果 521例孕妇中,IgG抗-A(B)效价大于或等于1∶128者有288例,占49.66%。其中IgG抗-A效价大于或等于1∶128者有161例,占IgG抗-A总人数的54.03%,IgG抗-B效价大于或等于1∶128者有127例,占IgG抗-B总人数的45.04%。结论通过微柱凝胶免疫检测法检测孕妇血清血型抗体IgG效价能有助于诊断新生儿溶血病,指导临床干预措施。     

一例2N型遗传性血管性血友病家系的表型诊断和基因型分析

目的：探讨1个遗传性血管性血友病（von Willebrand disease,vWD）2N型家系的表型诊断和基因型分析结果,明确患者的发病机制。方法：对该家系的先证者和家系成员进行出血时间、活化部分凝血活酶时间、瑞斯托霉素诱导的血小板聚集（ristocetin induced platelet aggregation,RIPA）试验、血管性血友病因子（von Willebrand factor,vWF）瑞斯托霉素辅因子、vWF抗原、vWF活性测定、vWF胶原结合试验、凝血因子Ⅷ（coagulation factor Ⅷ,FⅧ）活性、vWF与FⅧ结合试验检测,并作出表型诊断。提取先证者的外周血基因组DNA,用PCR法扩增VWF基因和F8基因的所有外显子及侧翼序列,通过直接测序分析VWF基因和F8基因变异。结果：vWD家系先证者的活化部分凝血活酶时间和出血时间明显延长,血浆瑞RIPA试验、vWF瑞斯托霉素辅因子、vWF抗原、vWF活性和vWF胶原结合试验检测结果均正常,FⅧ活性下降,vWF与FⅧ的结合能力降低。对先证者进行基因测序,发现其VWF基因19号外显子存在c.2446C>T（p.Arg816Trp）错义突变,其儿子在该位点为杂合突变,而先证者及家系成员的F8基因未发现突变。结论：c.2446C>T（p.Arg816Trp）错义突变是导致该家系先证者发生2N型遗传性vWD的原因。     

Neutralization of antifactor VIII inhibitors by recombinant porcine factor VIII

BACKGROUND: Inhibitory antibodies to factor (F) VIII (FVIII inhibitors) present a major clinical challenge as a complication of hemophilia A and as acquired autoantibodies in non-hemophiliacs. Porcine FVIII is a potentially useful therapeutic agent because of its low crossreactivity with many inhibitors. Recombinant porcine FVIII (rpFVIII) is undergoing clinical trials in inhibitor patients. OBJECTIVES: The goals of this study were to neutralize human FVIII inhibitors in vitro with rpFVIII and to characterize the relationship between recovery of FVIII activity and the antiporcine FVIII inhibitor titer. METHODS: rpFVIII was added to 28 antihuman FVIII inhibitor plasmas and assayed either immediately or after a 2 h preincubation at 37 degrees C. The concentration of rpFVIII required for recovery of FVIII activity to 50% of normal (EC(50)) was determined by polynomial regression analysis and compared with the antiporcine FVIII inhibitor titer measured by Bethesda assay. RESULTS: Six of nine plasmas classified as non-crossreactive by Bethesda assay had detectable inhibitory activity in the FVIII recovery assay. Recovery was decreased following a 2 h preincubation compared with immediate assay for the 19 plasmas with detectable antiporcine FVIII inhibitors. There was a linear relationship between EC(50) and inhibitor titer for plasmas with antiporcine FVIII titers ranging from 0.6 to 10 BU per milliliter in both the immediate and the 2 h preincubation assays. CONCLUSION: A quantitative method for analysis of inhibitor neutralization in vitro has been developed, which may be useful for comparison with pharmacodynamic studies of rpFVIII in FVIII inhibitor patients and subsequent rational dosing in this patient population.     

Surveillance for factor VIII inhibitor development in the Canadian Hemophilia A population following the widespread introduction of recombinant factor VIII replacement therapy

In the Fall of 1994 the majority of Canadian Hemophilia A (Factor VIII (F.VII) deficiency) patients who were receiving replacement therapy were converted to Recombinant Factor VIII (rF.VIII) from plasma derived products. This decision was taken and funded by the Canadian Blood Agency following the advice of the Association of Hemophilia Centre Directors of Canada (AHCDC) who considered this to be the safest replacement therapy available. Although it was the considered opinion of the AHCDC that there was no evidence available to support the theoretical concern that rF.VIII may prove more immunogenic than plasma derived products, patient follow up included intensive surveillance of all patients converted for this complication. A central reference laboratory was established and plasma specimens obtained before and 6-12 months following conversion to rF.VIII were referred for evaluation for inhibitor development by the classical Bethesda Assay. By concensus of the referring centers a Bethesda Unit (BU)/activity of 0.5 or greater was considered to be clinically significant. No increase in the incidence of inhibitor development has been recorded in 478 patients followed for one year after conversion. This pattern has not changed in 339 of these patients followed for a further year. Of interest was the finding by the reference laboratory that 8.0% of patients had BU activity of 0.5 or greater before conversion to rF.VIII. Many of these individuals lost this activity after conversion to rF.VIII as did others who appeared to have developed inhibitory activity during the first year of follow up but became inhibitor free in the second year of therapy. Overall, the incidence of true inhibitor development, i.e. negative pre-/positive post-conversion to rF.VIII, in this population of Hemophilia A patients was 2-3% over 2 years. This is similar to the incidence in patients treated with plasma derived products. These data emphasize the need for rigorous baseline evaluation in such investigations and the heterogeneity of the inhibitor response as assessed by in vitro assay. It was concluded that, although no attempt was made to correlate these in vitro data with clinical observations including F.VIII recovery and survival, the use of rF.VIII in hemophiliacs previously treated with plasma derived products was not associated with an increase in F.VIII inhibitor development.     

非肿块型乳腺导管内癌超声特征及与临床、病理、免疫组化指标表达间的相关性

目的：观察非肿块型乳腺导管内癌（ductal carcinoma in situ,DCIS）患者的超声表现,探讨其与临床、病理学特征及免疫组化指标表达间的相关性。方法：回顾性分析我院68例非肿块型DCIS患者术前的临床资料及超声检查结果,并观察其术后病理特征及雌激素受体、孕激素受体、人表皮生长因子受体2（human epidermal growth factor receptor 2,HER2）、Ki-67指数等免疫组化指标表达情况。根据超声图像特征,将患者分为导管型（Ⅰ型）、腺体型（Ⅱ型）、钙化型（Ⅲ型）和结构紊乱型（Ⅳ型）4组,分析4组DCIS患者的超声表现与其临床病理特征及免疫组化指标之间的相关性。结果：68例DCIS患者中,导管型为21例（30.9%）,腺体型为10例（14.7%）,钙化型为31例（45.6%）,结构紊乱型6例（8.8%）。各型患者间在年龄、月经状态、临床症状、前哨淋巴结转移、雌孕激素受体及Ki-67指数表达方面差异均无统计学意义（P>0.05）;导管型中以病理组织学表现为低级别的单纯DCIS、免疫组化指标HER2阴性表达为多见（81.0%）（P<0.05）;钙化型中以HER2阳性表达为多见（64.5%）（P<0.05）。中低级别DCIS及HER2阴性表达与导管型之间存在相关性,而高级别DCIS及HER2阳性表达与钙化型之间存在相关性（P<0.05）。结论：非肿块型DCIS患者的部分超声征象与其病理特征及免疫组化指标表达情况间存在一定相关性。     

The Bacillus cereus containing sub-branch most closely related to Bacillus anthracis, have single amino acid substitutions in small acid-soluble proteins, while remaining sub-branches are more variable

Hoffmaster et al. [Hoffmaster AR, Ravel J, Rasko DA, Chapman GD, Chute MD, Marston CK, et al. Identification of anthrax toxin genes in Bacillus cereus associated with illness resembling inhalation anthrax. Proc Natl Acad Sci U S A 2004;101:8449-54; Hoffmaster AR, Hill KK, Gee JE, Marston CK, De BK, Popovic T, et al. Characterization of Bacillus cereus isolates associated with fatal pneumonias: strains are closely related to Bacillus anthracis and harbor B. anthracis virulence genes. J Clin Microbiol 2006;44:3352-60] phylogenetically divided Bacillus cereus strains into 10 branches by amplified fragment length polymorphism (AFLP) with Branch F including all Bacillus anthracis strains and pneumonia-causing strains of B. cereus. There are four sub-branches within Branch F, referred to here as F1-A, F1-B, F2-A and F2-B. The B. anthracis strains are found within sub-branch F1-B. Concerning, the currently available B. cereus pneumonia-causing isolates, one was found to categorize within sub-branch F1-B and two within F2-B. In the following work the sequence variation between B. cereus strains was determined by MALDI-TOF MS and MS-MS for each strain of B. cereus in Branch F. ESI-MS was performed on selected strains for confirmation. Small acid-soluble proteins (SASPs) of B. cereus strains found in F1-B showed a single amino acid substitution, while strains in the other three sub-branches were more variable generally showing one or two amino acid substitutions. The single substitutions always occurred in the C-terminus. Double substitutions occurred in both N and C termini. Of the pneumonia-causing strains, one exhibited a single amino acid substitution, while the other two exhibited a two amino acid substitution.     

Heparin neutralization is essential for accurate measurement of factor VIII activity and inhibitor assays in blood samples drawn from implanted venous access devices

The purpose of this study was to determine the prevalence and effect of heparin contamination in samples drawn from implanted venous access devices (VADs, ports) on factor VIII activity and Bethesda inhibitor assay (BU) and the efficacy of heparinase to neutralize heparin. Plasma samples containing 85, 45, and 2 U/dL factor VIII were spiked in vitro with heparin from 0 to 3 U/mL. Factor VIII activity was assayed with a one-stage clotting assay on paired samples before and after heparinase, 25 mg/mL plasma. Paired patient samples drawn from VADs were assayed for heparin concentration, factor VIII, and BU before and after heparinase. At all three concentrations of factor VIII in vitro, the addition of heparin at 0.12 to 0.25 U/mL decreased assayed factor VIII activity. Heparinase neutralized up to 2 U/mL heparin and resulted in accurate factor VIII determination. Of 105 VAD samples, 47 (45%) had heparin contamination >0.05 U/mL. Of 47 heparin-contaminated samples, 42 showed decreased factor VIII activity in before/after comparisons. False-positive BU results were detected in 6 of 47 heparin-contaminated samples. Heparin contamination occurs frequently in samples drawn from VADs and could increase costs through excessive factor concentrate use. We recommend that all VAD samples be pretreated with heparinase before the assay of factor VIII activity or Bethesda inhibitor titers.     

Poly(lactide-co-glycolide) microspheres containing bupivacaine: comparison between gamma and beta irradiation effects

The β- and γ-irradiation effects on stability of microspheres made of poly(lactide-co-glycolide) 50:50 copolymer (PLGA) containing bupivacaine (BU) were studied. Microspheres containing 10, 25, and 40% w/w, respectively, of BU were prepared by spray drying and irradiated in air with β- and γ-irradiation at a dose of 25 kGy. Morphology (atomic force microscopy, particle-size analysis), physico–chemical characteristics (DSC and FT-IR spectroscopy), drug content and in vitro dissolution profile of microspheres were all determined; the stability of irradiated microspheres was evaluated over a 9-month period. The decrease of BU content in γ-irradiated microspheres was almost always constant independent of the amount of BU per sample, therefore it was in inverse proportion to drug loading (range between 5 and 15%). BU release rate increased immediately after irradiation and increased slightly until 90 days of storage. As far as β-irradiated microspheres are concerned, BU content decreased in a significant way (≈3%) only in microspheres containing 10% w/w of BU. Immediately after irradiation, drug release rate in β-irradiated microspheres increased less than in the corresponding γ-irradiated microspheres, and it did not change further over the following storage period. BU-loaded microspheres have been shown to be more stable against β- than γ-irradiation. AFM revealed that the surface roughness of the irradiated microspheres increases depending on irradiation. As such, if a parameter is quantifiable, it is proposed as a marker of degradation due to ionizing radiation.     

β-arrestin2通过调控自噬水平在结肠炎中的作用

目的 研究β-抑制蛋白2 （β-arrestin2）是否通过调控自噬水平在结肠炎中发挥作用。方法 β-arrestin2野生型（WT）小鼠和β-arrestin2基因敲除（KO）小鼠各20只,随机分为4组,每组各10只,分别为β-arrestin2 WT对照组和实验组,β-arrestin2 KO对照组和实验组。实验组小鼠自由饮用3%葡聚糖硫酸钠7 d诱导急性溃疡性结肠炎,对照组给予无菌ddH₂O。观察并记录各组小鼠的疾病活动指数。收集小鼠结肠组织,免疫组织化学和蛋白免疫印迹法检测自噬相关蛋白微管相关蛋白1轻链3β（LC3B）、Beclin1的表达水平。在HCoEpiC细胞中,通过siRNA沉默β-arrestin2的表达,Earle's平衡盐溶液（EBSS）处理细胞以诱导细胞自噬的发生,检测LC3B表达水平。结果 β-arrestin2 WT实验组小鼠的结肠黏膜自噬相关蛋白表达水平上调,而β-arrestin2 KO实验组小鼠的结肠炎症程度明显改善,自噬相关蛋白LC3B-Ⅱ/Ⅰ表达水平下降（P < 0.05）,但Beclin1表达水平比较差异无统计学意义（P > 0.05）。在HCoEpiC细胞中沉默β-arrestin2的表达,EBSS处理后,LC3B-Ⅱ/Ⅰ表达水平下调。结论 在结肠炎中,β-arrestin2调控自噬水平,当β-arrestin2缺失时,可以通过抑制自噬改善结肠炎。     

血友病A伴抑制物一例及其免疫耐受诱导治疗探讨

目的 探讨血友病A伴抑制物的免疫耐受诱导(immune tolerance induction,ITI)治疗,提高血友病A伴抑制物患者的诊疗水平.方法 对重型血友病A患者用APTT标准曲线一期法测定凝血因子Ⅷ(FⅧ)活性(FⅧ∶C);用Bethesda法定量测定FⅧ抗体;用长距离PCR方法检测内含子22倒位.结果 经检测发现患者为FⅧ基因22内含子倒位;ITI治疗3个月后,患者FⅧ抑制物滴度下降为0,输注FⅧ后回收率＞66%,治疗6个月后达到抑制物清除标准,停止治疗.结论 该患者是国内首例采用ITI成功治疗血友病A伴抑制物的病例,ITI是目前最有希望根除血友病抑制物的方法.