ANA和ENA检测在自身免疫性疾病中的应用评价

探讨抗核抗体和抗可溶性核抗原抗体的不同检测方法对自身免疫性疾病诊断的指导意义。回顾性分析2007年1月至2009年10月间在长征医院就诊的患者血清自身抗体的检测结果,549位患者,其中自身免疫病患者224例,非自身免疫病325例,所有患者血清同时检测ANA和ENA。以Hep-2细胞/肝组织为基质的间接免疫荧光法检测ANA,免疫印迹法检测ENA。两种检测方法产生4种检出模式:ANA+/ENA+、ANA-/ENA-、ANA+/ENA-和ANA-/ENA+。前两种模式共占检测的62.84%。ANA和ENA在自身免疫病患者中的阳性率(63.4%和58.5%)显著高于非自免病患者(16.9%和25.2%),ANA和ENA在自身免疫病组和非自身免疫病组间阳性率比较,差异有统计学意义(P<0.01)。两检测结果仅在MCTD和SLE患者中存在相关性(P<0.01),在其他观察组中不存在相关性(P>0.05)。间接免疫荧光法相对费时,而且需要操作者具备一定的经验,作为初筛实验,ANA的检测较ENA有更高的灵敏度,两者联合检测则将有利于提高检测的灵敏度和可靠性。     

流式荧光免疫技术检测抗核抗体谱的性能与临床应用评价

目的 分析流式荧光免疫技术(FFIA)检测自身抗体谱的结果及评价其在系统性红斑狼疮(SLE)诊断中的应用价值。方法 用流式荧光免疫法和免疫印迹法同时检测219例自身免疫病患者(包括92例SLE患者和127例非SLE的其他自身免疫病患者)、231例其他疾病患者、50例健康体检者血清的特异性自身抗体(抗nRNP、Sm、SS-A、SS-B、Jo-1、Scl-70、rRNP、PCNA、Nucleosome、Histone、CenP-B、M2、dsDNA),计算两种方法的阴阳符合率,并采用Kappa检验评估结果一致性;采用受试者工作特征曲线(ROC)比较各抗体在系统性红斑狼疮诊断中的应用价值。结果 FFIA与免疫印迹法检测各项自身抗体总符合率在87%至98%之间,Kappa值在0.179至0.854之间,其中抗CenP-B抗体一致性最好(Kappa值=0.854),抗His抗体一致性最差(Kappa值=0.179)。在219例自身免疫病例组数据中,根据试剂盒给定的阳性结果判断标准得到的免疫印记法灵敏度(62.0%)高于流式荧光(44.6%),而特异性略低于后者,分别为89.8%和94.4%,但二...     

自身免疫性肝病患者自身抗体检测及临床意义

目的 探讨自身免疫性肝病患者血清中出现的自身抗体等免疫学指标及临床意义.方法 对3 500例肝功能反复异常的患者采用间接免疫荧光法检测抗核抗体(ANA)、抗平滑肌抗体(SMA)、抗线粒体抗体(AMA).并对AMAM2型及抗可溶性肝抗原/肝胰抗原(抗SLA/LP)、抗肝肾微粒体抗体Ⅰ型(抗LKM-1)和抗肝特异性胞浆抗原Ⅰ型抗体(抗LC-1)等肝脏疾病相关的自身抗体进行检测.结果 3 500例患者中,自身免疫性肝炎患者29例,检出率为0.83%,其中符合Ⅰ型、Ⅱ型、Ⅲ型自身免疫性肝炎的比例占72.4%、10.3%和17.2%.原发性胆汁性肝硬化(PBC)患者58例,检出率为1.65%,血清中AMAM2型抗体阳性率为93.1%,其中19例AMAM2阳性患者进行肝穿病理检查时12例(63.7%)患者病理提示符合PBC诊断.结论 每种自身免疫性肝病都具有特征性自身抗体谱,注重自身抗体检测对明确诊断及鉴别诊断自身免疫性肝病具有重要的临床意义.     

免疫印迹法与免疫荧光法检测寻常型天疱疮自身抗体的比较

比较免疫印迹法与免疫荧光法对检测寻常型天疱疮抗体的临床应用价值。利用重组寻常型天疱疮抗原 (PVAEC1 2 )进行寻常型天疱疮 (PV )患者的免疫印迹法 (IBT )检测 ,并同时应用直接免疫荧光法 (DIF )和间接免疫荧光法 (IIF )检测 2 5例PV患者。IBT 2 1例阳性 (84 % ) ,DIF 2 0例阳性 (80 % ) ,IIF 18例阳性 (72 % )。正常对照 10例均为阴性。免疫荧光技术在PV的诊断与鉴别诊断中有其重要作用。利用重组PVAEC1 2行IBT以检测PV患者血清中抗PVA抗体 ,与免疫荧光法敏感性无统计学差异     

抗核抗体在自身免疫性疾病的检测及应用

自身免疫性疾病是机体的自身免疫应答失控,反应过度造成机体的正常结构成分被相应的抗体或具有免疫功能的细胞作用后引起器质性病变及功能性障碍的状态。自身免疫性疾病病人血循环中存在多种自身抗体,其中抗核抗体（ANA）、抗双链脱氧核糖核酸（抗dsDNA）、抗核抗原（抗ENA）三项联检为自身免疫性疾病的诊断、治疗及预后提供了重要依据。我们对198例自身免疫性疾病病人的血清进行了ANA、抗dsDNA、抗ENA的检测,并对这些自身抗体与自身免疫性疾病各疾病问的关系进行了分析,现报告如下。     

系统性红斑狼疮患者自身抗体检测及意义

目的：探讨自身抗体在系统性红斑狼疮（SLE）中的意义。方法：采用放射免疫分析检测50例SLE患者血清中的抗双链DNA（dsDNA）抗体,采用德国IMTEC公司抗核抗体谱线性印迹法检测抗核小体抗体（AnuA）、抗组蛋白抗体（AHA）、抗StuD1等其他自身抗体。结果：抗StuD1阳性率最高（82％）,其次是抗R060KD抗体（80％）和AunA（72％）;抗dsDNA、AHA、抗U1-RNP、抗R052KD、抗SSB的阳性率分别为44％、32％、58％、48％、24％。其他三种自身抗体阳性率很低。结论：SLE患者血清中可出现大量的自身抗体,同时检测多种自身抗体能提高对SLE的鉴别诊断。     

58例原发性胆汁性肝硬化患者自身抗体特征分析

目的：探讨自身抗体对原发性胆汁性肝硬化（PBC）患者的诊断应用价值。方法：用间接免疫荧光、免疫印迹法检测了58例PBC患者抗核抗体（ANA）、抗平滑肌抗体（SMA）、抗线粒体抗体（AMA），并对AMAM2亚型及抗可溶性肝抗原/肝胰抗原（SLA/LP）、抗肝肾微粒体Ⅰ型（LKM-1）和抗肝特异性胞浆Ⅰ型抗体（LC-1）等肝脏疾病相关的自身抗体进行检测。结果：PBC患者自身抗体以AMA和AMAM2亚型为主。其阳性率分别为96.5%和93.1%。患者的抗体滴度均大于1∶100，其中有8例出现ANA和SMA，1例出现AMA和SLA/LP同时阳性，表现与Ⅰ型和Ⅲ型自身免疫性肝炎重叠。另有19例AMAM2阳性患者进行肝穿病理检查时，12例（63.7%）患者病理提示符合PBC诊断。结论：自身抗体对PBC有诊断意义，注重自身抗体的检测对明确自身免疫性肝病有重要的临床意义。     

肝抗原自身抗体在自身免疫性肝病诊断中的意义

目的 探讨肝抗原自身抗体在自身免疫性肝病患者血清中的阳性率。方法 将患者分为3组：①自身免疫性肝病组，包括自身免疫性肝炎(AIH)、原发性胆汁性肝硬化(PBC)，原发性硬化性胆管炎(PSC)。②各类病毒性肝炎组；③不明原因肝损伤组。分别用间接免疫荧光法、免疫印迹法检测肝抗原(SAL/LP、LKM-1、LC-1、AMA-M2)自身抗体。结果 抗SLP／LP抗体在AIH患者血清中阳性率为46．4％，明显高于LKM-1(13．3％)、LC-1(0．O％)及AMA-M2(13．3％)抗体，并且在病毒性肝炎患者血清中呈阴性反应。抗AMA-M2抗体在PBC患者血清中阳性率达95．0％。不明原因肝损伤组患者中有10．0％的AMA-M2抗体阳性。结论 抗SLA/LP抗体对AIH具有特异性，肝抗原自身抗体的检测将有助于自身免疫性肝病患者的诊断及治疗。     

83例原发性肝癌患者自身抗体检测结果分析

目的:探讨原发性肝癌患者自身抗体产生的规律及临床意义.方法:以临床诊断为原发性肝癌患者83例作为研究对象,采用间接免疫荧光法及免疫印迹法检测患者血清中自身抗体.结果:83例肝癌患者自身抗体检出率为37.3%(31/83),其中HBV感染患者检出率35.9%(23/64),HCV感染者检出率50.0%(8/16),二者差异无统计学意义(P＞0.05).能检测到一种、两种及三种自身抗体的患者分别为24.1%、12.1%和1.2%.抗核抗体(ANA)、抗细胞骨架抗体(CS)、抗平滑肌抗体(SMA)和抗线粒体抗体(AMA)的检出率分别为24.1%、13.3%、9.6%和4.8%.抗核抗体谱分析有Ro-52、CENP-B(着丝点B抗原)等自身抗体阳性.AFP(甲胎蛋白)异常及AFP正常的患者自身抗体检出率分别为36.8%和22.2%,差异无统计学意义(P＞0.05).HBV-DNA阳性与HBV-DNA阴性患者自身抗体检出率分别为40.5%和 27.3%,差异无统计学意义.女性患者自身抗体检出率明显高于男性,差异有统计学意义.结论:37.3%的原发性肝癌患者能检测到非器官特异性的自身抗体:ANA、CS、SMA和AMA,大多患者以检出其中一种抗体为主,部分患者能检出二至三种自身抗体.ANA、CS、SMA和AMA四种自身抗体中检出率最高的是ANA,其次是CS和SMA,检出率最低的是AMA.各抗体均以低滴度为主.ANA荧光模型以颗粒型为主.血清AFP含量的高低与自身抗体检出率之间未见明显相关性.HBV-DNA阳性患者与HBV-DNA阴性患者自身抗体检出率差异无统计学意义.女性患者自身抗体检出率明显高于男性.     

重组多肽酶联免疫吸附法检测抗LKM-1抗体的初步应用

目的：制备人自身抗原细胞色素P4502D6（CYP2D6）257-351位氨基酸片段融合蛋白作为自身抗原,探讨ELISA检测抗LKM-1抗体的敏感性和特异性。方法：以肝脏的cDNA混合文库为模板作PCR,将PCR产物与真核表达载体pEGH共同转化酿酒酵母Y258,碱裂解法进行质粒制备,PCR扩增鉴定。表达载体构建成功后,在半乳糖的诱导下表达产生重组融合蛋白,经GST亲和层析法进行纯化后,免疫印迹法鉴定抗原性,ELISA检测抗LKM-1抗体阳性血清及部分其他结缔组织病患者血清中的抗LKM-1抗体。结果：重组融合蛋白在宿主菌中获得表达,免疫印迹法鉴定表明其能与标准抗LKM-1抗体阳性血清反应,而与正常血清、其他抗血清无反应。在26份抗LKM-1抗体阳性血清中,6份抗HCV抗体阳性血清用重组多肽ELISA检测有5份呈阳性,其余20份血清用重组多肽ELISA检测均呈阳性；20份其他结缔组织病患者血清用重组多肽ELISA检测均为阴性。结论：重组的257-351位氨基酸片段是CYP2D6抗原的主要抗原表位区域,以重组多肽为基质ELISA检测抗LKM-1抗体的敏感性较高,为进一步研究抗体水平与临床病情变化的相关性奠定了基础。     

Pain and Effusion and Quadriceps Activation and Strength

Context:

Quadriceps dysfunction is a common consequence of knee joint injury and disease, yet its causes remain elusive.

Objective:

To determine the effects of pain on quadriceps strength and activation and to learn if simultaneous pain and knee joint effusion affect the magnitude of quadriceps dysfunction.

Design:

Crossover study.

Setting:

University research laboratory.

Patients or Other Participants:

Fourteen (8 men, 6 women; age = 23.6 ± 4.8 years, height = 170.3 ± 9.16 cm, mass = 72.9 ± 11.84 kg) healthy volunteers.

Intervention(s):

All participants were tested under 4 randomized conditions: normal knee, effused knee, painful knee, and effused and painful knee.

Main Outcome Measure(s):

Quadriceps strength (Nm/kg) and activation (central activation ratio) were assessed after each condition was induced.

Results:

Quadriceps strength and activation were highest under the normal knee condition and differed from the 3 experimental knee conditions (P < .05). No differences were noted among the 3 experimental knee conditions for either variable (P > .05).

Conclusions:

Both pain and effusion led to quadriceps dysfunction, but the interaction of the 2 stimuli did not increase the magnitude of the strength or activation deficits. Therefore, pain and effusion can be considered equally potent in eliciting quadriceps inhibition. Given that pain and effusion accompany numerous knee conditions, the prevalence of quadriceps dysfunction is likely high.Key Words: arthrogenic muscle inhibition, central activation failure, voluntary activation, muscles

Key Points

• Knee pain and effusion resulted in arthrogenic muscle inhibition and weakness of the quadriceps.
• The simultaneous presence of pain and effusion did not increase the magnitude of quadriceps dysfunction.
• To reduce arthrogenic muscle inhibition and improve muscle strength, clinicians should employ interventions that target removing both pain and effusion.

Quadriceps weakness is a common consequence of traumatic knee joint injury^1,2 and chronic degenerative knee joint conditions.^3,4 Arthrogenic muscle inhibition (AMI), a neurologic decline in muscle activation, results in quadriceps weakness and hinders rehabilitation by preventing gains in strength.⁵ The inability to reverse AMI and restore muscle function can lead to decreased physical abilities,⁶ biomechanical deficits,⁷ and possibly reinjury.⁵ Furthermore, researchers^8,9 have suggested that quadriceps weakness resulting from AMI may place patients at risk for developing osteoarthritis in the knee. In light of the substantial influence of quadriceps AMI on these clinically relevant outcomes, we need to improve our understanding of the factors that contribute to this neurologic decline in muscle activity so efforts to target and reverse it can be implemented and gains in strength can be achieved more easily.Joint injury and disease are accompanied by numerous sequelae (ie, pain, swelling, tissue damage, inflammation), so ascertaining which one ultimately leads to neurologic muscle dysfunction is difficult. Whereas a joint effusion can result in AMI,^10–12 the effects of pain are less understood despite many clinicians attributing AMI to pain. Using techniques that introduce knee pain without accompanying injury may provide insights into the role of pain in eliciting AMI.The degree of knee joint damage may play a role in the quantity of AMI that manifests. Hurley et al^13,14 demonstrated that quadriceps AMI, measured using an interpolated-twitch technique, was greater in patients with extensive traumatic knee injury (eg, fractured tibial plateau, ruptured medial collateral ligament, and medial meniscectomy) than patients with isolated joint trauma (ie, isolated anterior cruciate ligament [ACL] rupture). Similarly, patients with more knee joint symptoms (ie, greater number of symptoms and increased severity of symptoms) may present with greater magnitudes of quadriceps inhibition. Recently, investigators¹⁵ have suggested that patients with more pain display less quadriceps strength, supporting this tenet. Given that effusion and pain often present simultaneously with joint injuries and diseases, such as ACL injury and osteoarthritis, examining both the isolated and cumulative effects of these sequelae appears warranted to determine if they influence the magnitude of muscle inhibition.Experimental joint-effusion and pain models are safe and effective experimental methods that allow for the isolated examination of their effects on muscle function. The effusion model, whereby sterile saline is injected directly into the knee joint capsule,⁷ produces a clinically relevant magnitude of the joint effusion that may be present with traumatic injury. Effusion is thought to activate group II afferents responding to stretch or pressure,^16–18 which in turn may facilitate group Ib interneurons and result in quadriceps AMI.⁵ The pain model involves injecting hypertonic saline into the infrapatellar fat pad to produce anteromedial knee pain similar to that described in patients with patellofemoral pain syndrome.¹⁹ Pain is considered to initiate AMI through activation of group III and IV afferents that act as nocioceptors to signal damage or potential damage to joint structures.^16–18 The firing of these afferents then may lead to facilitation of group Ib interneurons, the flexion reflex, or the gamma loop, ultimately resulting in quadriceps inhibition.²⁰ Thus, these models allow us to create symptoms that are associated with knee injury and have the added benefit of providing a way to examine their effects in isolation.Therefore, the purpose of our study was to determine the effects of pain on quadriceps strength and activation and to learn if simultaneous pain and knee joint effusion would affect the magnitude of quadriceps dysfunction. We hypothesized that pain alone would result in quadriceps inhibition and that the magnitude of inhibition would be greater when effusion and pain were present simultaneously.     

即早基因c-fos与脑血管病及学习记忆

即早基因c-fos是广泛存在于原核细胞和真核细胞的高度保守基因.在正常情况下,c-fos基因参与细胞生长、分化、信息传递、学习和记忆等生理过程,而在病理情况下c-fos基因表达及调控变化与多种疾病的发生和发展有关.C-fos在中枢神经系统的某些部位可有基础水平的表达,但表达很低,当受到如脑缺血、脑出血、痫性发作、应激等刺激后,其在数十分钟内做出反应,在对外界刺激-转录耦联的信忠传递过程中起着核内第三信使的重要作用.     

Opportunities and challenges in the prevention and control of cancer and other chronic diseases: children's diet and nutrition and weight and physical activity

OBJECTIVE: The purpose of this article is to review the role of behavioral research in disease prevention and control, with a particular emphasis on lifestyle- and behavior-related cancer and chronic disease risk factors--specifically, relationships among diet and nutrition and weight and physical activity with adult cancer, and tracking developmental origins of these health-promoting and health-compromising behaviors from childhood into adulthood. METHOD: After reviewing the background of the field of cancer prevention and control and establishing plausibility for the role of child health behavior in adult cancer risk, studies selected from the pediatric published literature are reviewed. Articles were retrieved, selected, and summarized to illustrate that results from separate but related fields of study are combinable to yield insights into the prevention and control of cancer and other chronic diseases in adulthood through the conduct of nonintervention and intervention research with children in clinical, public health, and other contexts. RESULTS: As illustrated by the evidence presented in this review, there are numerous reasons (biological, psychological, and social), opportunities (school and community, health care, and family settings), and approaches (nonintervention and intervention) to understand and impact behavior change in children's diet and nutrition and weight and physical activity. CONCLUSIONS: Further development and evaluation of behavioral science intervention protocols conducted with children are necessary to understand the efficacy of these approaches and their public health impact on proximal and distal cancer, cancer-related, and chronic disease outcomes before diffusion. It is clear that more attention should be paid to early life and early developmental phases in cancer prevention.