寻常型天疱疮抗原Dsg3 EC1-EC2片段的重组表达

寻常型天疱疮（Pemphigus vulgaris,PV）是一种自身免疫性皮肤病。靶抗原位于细胞间重要的连接结构-桥粒上，主要是桥粒芯糖蛋白3（desmoglein3，Dsg3），获得Dsg3，不仅可以用来检测PV抗体，还可用来进行基础研究，探索天疱疮的发病机理。     

大疱性类天疱疮患者血清中抗BP180 IgE、抗BP230 IgE和总IgE间的相关性

目的检测大疱性类天疱疮(BP)患者外周血嗜酸粒细胞、血清总IgE、抗BP180和抗BP230的IgE抗体,并分析它们之间的相关性。方法用ELISA方法检测63例BP患者血清的总IgE抗体、抗BP180 IgE抗体和抗BP230IgE抗体水平,并分析患者外周血嗜酸粒细胞水平、BPDAI评分与IgE抗体的相关性。结果 63例BP患者中,总IgE阳性者41例,阳性率65.08%;抗BP180 IgE阳性者37例,阳性率58.73%;抗BP230 IgE抗体阳性者47例,阳性率74.60%。总IgE和抗BP230 IgE呈正相关(r=0.643,P0.001)。63例患者中共34例记录了外周血嗜酸粒细胞水平,其中19例BP患者外周血嗜酸粒细胞数量增多(55.88%)。BP180 IgE抗体与BPDAI中的红斑评分呈正相关(r=0.934,P0.05)。结论 IgE在BP致病过程中发挥作用,IgE自身抗体可能主要与BP230相结合,同时抗BP180 IgE可能会引发BP患者红斑。     

Dsg3刺激诱导天疱疮患者一级亲属T细胞活化的研究

目的 研究在特异抗原桥粒芯糖蛋白3(desmoglein3,Dsg3)刺激下寻常型天疱疮(PV)患者一级亲属淋巴细胞Th1/Th2、Tc1/Tc2极化状态的变化,探讨PV发生机制.方法 PV患者一级亲属外周血单个核细胞(PBMC)在Dsg3刺激下体外培养3 d,流式细胞仪四色胞内染色分析PV患者一级亲属CD4~+ T细胞和CD8~+T细胞中IFN-γ~+和IL-4~+细胞的百分率,观察PV患者一级亲属Th1/Th2、Tc1/Tc2比例的变化.结果 PV抗体阳性一级亲属中的PBMC经Dsg3刺激培养者与未经Dsg3刺激者比较,Th2、Tc2百分率均显著升高(10.13%±3.72%vs 7.28%±3.58%,20.01%±10.43% vs 14.91%±8.06%,P<0.05);经Dsg3刺激培养后PV抗体阳性一级亲属组与正常对照组比较Th2、Tc2百分率均显著升高(10.13%±3.72%VS 6.10%±2.82%,20.01%±10.43% vs9.58%±5.49%,P<0.05).结论 特异性抗原Dsg3刺激下,自身反应性T细胞活化,Th1/Th2、Tc1/Tc2细胞分化失衡,可能在PV的启动阶段发挥重要作用.     

寻常型天疱疮的Dsg3特异性抗体反应及其基因限制性

探讨寻常型天疱疮中的自身抗原桥粒芯蛋白（Dsg3)特异性抗体反应及其基因限制性，为自身免疫性疾病机制的研究提供依据。采用RT－PCR法克隆自身抗原Dsg3E1,E2,E3,E4,E5多肽片段的cDNA，定向插入表达载体PGEX－2T，导入大肠杆菌JM109中表达重组融合蛋白并经GST层析柱纯化，进一步经免疫印迹法与PV患者阳性血清反应；应用序列特异性引物聚合酶链式反应（SSP－PCR）技术对HLA－Ⅱ类等位基因进行特异性体外扩增，分析了天疱疮患者HLA基因的DR位点的DRB1、DQB1多态性。Dsg3 E1,E2和E4可与PV患者阳性血清反应，而不与疾病对照组、正常对照组反应。在10个PV患者中，均携带HLA的DR4或／和DR14抗原，有7个DRB1*0402,4个DRB1*1401,7个DQB1*0302,4个DQB1*0503.Dsg3 E1,E2和E4中包含抗体反应相关的抗原表位，HLA DRB1*0402t DQB1＊0302与PV发病中抗E4抗体反应密切相关。     

寻常型天疱疮自身抗原Dsg3片段的重组表达和特异性细胞反应

目的 :探讨寻常型天疱疮自身抗原Dsg3在特异性T细胞反应中的作用 ,为自身免疫性疾病机制的研究提供依据。方法 :根据Genbank中的Dsg3序列分析 ,采用RT PCR法克隆自身抗原Dsg3E1,E2 ,E3,E4,E5多肽片段的cDNA ,定向插入表达载体PGEX 2T ,导入大肠杆菌JM10 9中表达重组融合蛋白并经GST层析柱纯化 ;进一步与PV患者及疾病对照组、正常对照组T细胞混合培养 ,观察T细胞增殖反应。结果 :Dsg3E1,E2和E4,E5可刺激PV患者T细胞反应 ,而不与疾病对照组、正常对照组反应。结论 :Dsg3E1,E2和E4,E5中包含T B细胞作用相关的抗原表位 ,在PV发病中起重要作用。     

导常型天疱疮自身抗原Dsg3片段的重组表达和特异性细胞反应

目的：探讨寻常型天疱疮自身抗原Dsg3在特异性T细胞反应中的作用，为自身免疫性疾病机制的研究提供依据。方法：根据Genbank中的Dsg3序列分析，采用RT-PCR法克隆自身抗原Dsg3E1,E2,E3,E4,E5多肽片段的cDNA,定向插入表达载体PGEX-2T，导入大肠杆菌JM109中表达重组融合蛋白并经GST层析柱纯化；进一步与PV患者及疾病对照组、正常对照组T细胞混合培养，观察T细胞增殖反应。结果：DsgE1，E2和E4，E5可刺激PV患者T细胞反应，而不与疾病对照组、正常对照组反应。结论：Dsg3E1,E2和E4，E5中包含T-B细胞作用相关的抗原表位，在PV发病中起重要作用。     

系统性红斑狼疮患者血清miR-23b、CCL19水平与疾病活动度及补体水平的相关性分析

目的 探究系统性红斑狼疮(SLE)患者血清微小RNA-23b(miR-23b)、趋化因子配体19(CCL19)水平与疾病活动度、补体C3、补体C4的相关性.方法 选取2019年1月至2021年1月在本院风湿免疫科初诊、尚未治疗的SLE患者176例为SLE组,同期体检健康者124例为健康对照组.比较两组血清miR-23b、CCL19、补体C3、补体C4水平;采用SLE疾病活动指数2000(SLEDAI-2000)评分评估SLE患者的疾病活动度,比较不同活动度组血清指标水平;并分析miR-23b、CCL19与疾病活动度、补体C3、补体C4的相关性以及血清指标对SLEDAI-2000评分的影响.结果 SLE组血清miR-23b、补体C3、补体C4水平低于对照组,血清CCL19水平高于对照组,差异有统计学意义(P<0.05).血清miR-23b、补体C3、补体C4水平在重度活动组<中度活动组<轻度活动组<无活动组,血清CCL19水平在重度活动组>中度活动组>轻度活动组>无活动组,差异有统计学意义(P<0.05).Pearson相关性分析显示,miR-23b与补体C3、补体C4呈正相关(r=0.616、0.712,P<0.05),CCL19与补体C3、补体C4呈负相关(r=-0.342、-0.516,P<0.05).Spearman等级相关分析显示,miR-23b与疾病活动度呈负相关(r=-0.785,P<0.05),CCL19与疾病活动度呈正相关(r=0.740,P<0.05).多元线性回归分析显示,miR-23b、CCL19、补体C3、补体C4是SLEDAI-2000评分的影响因素(P<0.05);miR-23b、补体C3、补体C4负向影响SLEDAI-2000评分(B=-5.750、-2.854、-19.490,95％CI=-8.075～-3.425、-5.217～-0.492、-25.431～-13.550,均P<0.05),CCL19正向影响SLEDAI-2000评分(B=0.030,95％CI=0.025～0.036,P<0.05).结论 SLE患者血清miR-23b水平降低、CCL19水平升高,miR-23b、CCL19与疾病活动度、补体C3、补体C4存在相关性,且miR-23b、CCL19、补体C3、补体C4共同影响疾病活动度.     

NSPs在系统性红斑狼疮患者血清中的表达及临床意义北大核心CSCD

目的:探讨血清中3种主要中性粒细胞丝氨酸蛋白酶(NSPs)NE、CG、PR3在系统性红斑狼疮(SLE)患者中的表达及临床意义。方法:ELISA测定66例SLE患者、39例健康对照者血清NE、CG、PR3水平,分析SLE患者NE、CG、PR3水平与病情活动度的相关性,并分析SLE患者NE、CG、PR3水平与实验室指标及临床表现的关系。结果:SLE组血清NE、CG、PR3水平均高于健康对照组(P<0.05)。相关性分析显示,SLE患者血清NE、CG、PR3水平与SLE疾病活动评分呈正相关(r=0.420,0.269,0.475,P<0.05)。血清NE水平升高的SLE患者C4水平更低,疾病活动评分更高,抗核小体抗体及抗核糖体P蛋白抗体阳性率升高;血清CG水平升高的SLE患者肾损伤和血液学异常发生率、抗dsDNA抗体和抗核小体抗体阳性率升高,疾病活动评分更高,C3、C4水平更低;血清PR3水平升高的SLE患者关节炎发生率更高(P<0.05)。结论:SLE患者血清NSPs水平较健康对照组显著升高,且与SLE疾病活动程度呈正相关,NSPs可能参与SLE疾病发生发展过程。     

负性调控因子IRAKM 在系统性红斑狼疮免疫调节异常中的作用

目的:研究白介素-1 受体相关激酶-M(IRAKM)在系统性红斑狼疮(SLE)中的表达及其与免疫调节的关系。方法:将103 例SLE 患者根据其疾病活动度评分(SLEDAI)分为活动期组(55 例)和稳定期组(48 例),另选40 名体检健康者作为对照组;采用实时荧光定量PCR(RT-PCR)法检测3 组外周血单核细胞中IRAKM mRNA 的表达量,酶联免疫吸附法(ELISA)检测血清中抗ds-DNA 抗体、抗Sm 抗体水平,免疫散射比浊法测定血清补体C3、C4 水平;应用单因素方差分析法分析3 组各观察指标的差异,Pearson 或Spearman 分析IRAKM 与SLE 自身抗体及补体的相关性。结果:活动期组和稳定期组SLE 患者的IRAKM mRNA 表达量均明显低于对照组(P<0.05),且活动期组比稳定期组更低,差异有统计学意义(P<0.05);与对照组比,活动期组和稳定期组血清抗ds-DNA 抗体和抗Sm 抗体水平均明显升高(P<0.05),补体C3、C4 的水平均明显降低(P<0.05),且活动期组血清自身抗体和补体水平变化较稳定期组更显著(P<0.05);SLE 患者的IRAKM mRNA 表达量与抗ds-DNA 抗体和抗Sm 抗体水平呈负相关(P<0.05),与补体C3、C4 水平呈正相关(P<0.05)。结论:IRAKM 负性调控参与SLE的免疫调节过程,其表达水平与SLE 病情活动程度密切相关。     

血清抗C1q抗体在系统性红斑狼疮疾病活动性判断及狼疮肾炎诊断中的价值

目的探讨抗C1q抗体(C1qAb)在系统性红斑狼疮(SLE)活动性及狼疮肾炎(LN)诊断和疾病活动性判断中的价值。方法采用酶联免疫吸附法检测SLE患者(n=89)、疾病对照组(n=56)和正常对照组(n=42)血清中的抗C1q抗体阳性率,并与SLE患者临床实验室指标﹑活动性评分进行分析。结果 C1qAb的阳性率在SLE患者中显著高于疾病对照组和正常对照组患者(P<0.05);C1qAb阳性的SLE患者肾损发生率、活动性狼疮发生率及抗dsDNA抗体的阳性率均高于C1qAb阴性患者(P<0.05);C1qAb与SLEDAI活动性评分、抗核小体抗体(anti-nucleosome antibody,AnuA)及抗dsDNA抗体呈正相关(P<0.05)。结论抗C1q抗体对SLE的诊断和疾病活动性判断有重要价值;抗C1q抗体参与了SLE肾脏损害的发病机制。     

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.