不同类型HBV感染者外周血Treg/Th17细胞的变化及意义

目的:探讨不同类型HBV感染者外周血中CD4+ Foxp3+ Treg/Th17细胞的变化及意义.方法:选取15例急性乙型肝炎(Acute Hepatitis B,AHB)患者、40例慢性乙型肝炎(Chronic hepatitis B,CHB)患者、40例无症状携带者(Asymptomatic HBV carriers,AsC)及30例健康对照者,分别采用流式细胞术、RT-PCR和ELISA检测外周血CD4+ Foxp3+ Treg/Th17细胞百分率、核转录因子foxhead winged-helix box protein 3 (Foxp3)/retinoid-related orphan receptor gamma-t (RORγt) mRNA的表达以及血浆转化生长因子-β1(Transforming growth factor-31,TGF-β1)/IL-17的水平.结果:AHB组患者CD4+ Foxp3+ Treg/CD4+T细胞百分率、Foxp3 mRNA及TGF-β1水平与正常对照组相比无明显差异(P＞0.05);而CD4+ IL-17 +/CD4+T细胞百分率、RORγtmRNA及IL-17水平与正常对照组相比明显升高,差异有统计学意义(P＜0.05).CHB组患者CD4+ Foxp3+ Treg/CD4+T细胞百分率、Foxp3 mRNA、TGF-31水平及CD4+ IL-17 +/CD4+T细胞百分率、RORγt mRNA、IL-17水平与正常对照组相比均明显升高,差异有统计学意义(P＜0.05);与正常对照组相比,AsC组患者CD4+ Foxp3+ Treg/CD4+T细胞百分率、Foxp3 mR-NA、TGF-β1水平及CD4+ IL-17 +/CD4+T细胞百分率、RORγt mRNA、IL-17水平无明显差异(P＞0.05).结论:在不同类型HBV感染者外周血中Treg/Th17细胞失衡,Treg/Th17细胞可能与HBV感染的状态有关.     

急性冠脉综合征患者外周血中Foxp3~+调节性T细胞的变化

目的:探讨急性冠脉综合征(ACS)患者外周血Foxp3+(包括CD4+Foxp3+和CD4+CD25+Foxp3+)调节性T细胞(Treg)的变化与意义。方法:分别采用流式细胞术(FCM)、实时定量PCR和酶联免疫吸附法(ELISA)检测44例ACS患者,20例稳定性心绞痛(SA)患者和24例对照组患者外周血Treg细胞百分率,转录因子Foxp3的mRNA表达和血浆TGF-β1的浓度。结果:与对照组患者和SA组患者相比,ACS组患者外周血中CD4+Foxp3+、CD4+CD25+Foxp3+Treg细胞的百分率,Foxp3 mRNA的表达和血浆TGF-β1浓度明显降低(P<0.05),而CD4+CD25+Treg细胞的百分率在三组之间并无显著性差异。结论:ACS患者外周血Foxp3+Treg数量和/或功能的下调,Foxp3+Treg的变化可能与斑块的不稳定密切相关。     

卵巢癌患者外周血Th17/Treg细胞的检测及其临床意义

目的:检测卵巢癌患者外周血中辅助性T细胞17(T help cell 17,Th17)/调节性T细胞(regulatory T cell,Treg)的水平并探讨其临床意义。方法:选取55例卵巢癌和60例健康对照者为研究对象,采用细胞内染色流式细胞术(Flow cytometry,FCM)、实时定量PCR(real time PCR)、酶联免疫吸附法(Enzyme linked immunosorbent assay,ELISA)法检测外周血Th17/Treg细胞百分率、核转录因子retinoid-related orphan receptor gamma-t(RORγt)/foxhead winged-helix box protein 3(Foxp3)mRNA的表达以及血浆中白细胞介素17(Interleukin-17,IL-17)/转化生长因子-β1(Transforming growth factor-β1,TGF-β1)的水平。结果:卵巢癌患者外周血中CD4+IL-17+/CD4+T细胞百分率、RORγt mRNA及IL-17水平与正常对照组相比明显升高,差异有统计学意义(P<0.05)。与正常对照组相比,卵巢癌患者外周血中CD4+Foxp3+Treg/CD4+T细胞百分率、Foxp3 mRNA及TGF-β1水平明显升高,差异有统计学意义(P<0.05)。结论:卵巢癌患者外周血中Th17/Treg细胞失衡,Th17/Treg细胞可能参与卵巢癌的发病过程。     

茯苓多糖对系统性红斑狼疮患者Th17/Treg平衡的影响

目的:研究茯苓多糖对系统性红斑狼疮(SLE)患者外周血辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡的免疫调节作用。方法:选取45例SLE患者和35例健康对照者,应用磁珠分选法分离外周血CD4~+ T细胞,流式细胞术检测CD4~+ T细胞中Th17和Treg细胞的比例。用茯苓多糖分别处理健康对照者及患者的CD4~+ T细胞,MTT法检测细胞活力以测定茯苓多糖毒性,ELISA检测细胞中白细胞介素17(IL-17)、IL-6、IL-10及转化生长因子β(TGF-β)的含量,RT-q PCR和Western blot法分别测定维甲酸相关孤儿受体γt(RORγt)与叉头框蛋白P3(Foxp3)的mRNA和蛋白表达水平。结果:与健康对照组相比,SLE患者的Th17细胞比例显著升高,Treg细胞比例明显降低(P0.05)。用100μg/L的茯苓多糖处理SLE患者CD4~+ T细胞,与空白对照组相比,IL-17和IL-6的含量显著降低,IL-10和TGF-β的含量明显上升(P0.05);RORγt的mRNA和蛋白表达显著下降,同时Foxp3的表达在mRNA和蛋白水平上明显增加(P0.05);并且Th17/Treg的比值降低(P0.05)。结论:茯苓多糖可以通过升高Treg并降低Th17细胞的比例,对SLE起到一定的治疗作用。     

支气管哮喘患者外周血Th17、CD4~+CD25~+Treg细胞表达特征

目的:探讨外周血Th17和CD4+CD25+调节性T细胞(Treg)在支气管哮喘患者中的表达特征。方法:41例慢性持续期哮喘患者,分为间歇-轻度组(n=23)和中重度组(n=18),行肺功能检查和哮喘控制问卷(ACQ)调查,20例正常人作为对照。通过流式细胞术检测外周血Th17和CD4+CD25+Treg细胞的比例。ELISA检测血浆以及植物血凝素刺激24小时后外周血单个核细胞(PBMC)上清液中的IL-17、IL-10、TGF-β水平。结果:中重度哮喘组外周血Th17细胞比例及血浆IL-17水平高于间歇-轻度哮喘和正常人组,而外周血CD4+CD25+Foxp3+Treg细胞比例及血浆IL-10、TGF-β水平则降低。中重度哮喘组PBMC上清液中IL-17水平增高。哮喘患者FEV1(%预计值)与Th17细胞及血浆IL-17表达成负相关,与CD4+CD25+Treg表达成正相关。ACQ平均得分与Th17细胞和血浆IL-17表达成正相关,与外周血CD4+CD25+Treg表达成负相关。结论:中重度哮喘中外周血Th17细胞应答增强,而CD4+CD25+Treg细胞缺乏,哮喘的严重程度及症状控制与外周血Th17/Treg免疫应答失衡密切相关。     

反复自然流产患者外周血Th17和Treg细胞相关细胞因子的测定

目的 通过检测反复自然流产(RSA)患者外周血Th17和Treg细胞特异性转录因子RORγt和Foxp3 mRNA的表达及IL-6、IL-17、TGF-β1的水平,探讨Th17和Treg细胞在RSA发病中的作用.方法 选取2011-12/2012-04郑州大学第三附属医院50例RSA患者为研究对象,同期选取50名健康早孕女性作为对照组.采用实时荧光定量PCR检测外周血RORγt和Foxp3 mRNA的表达水平,并分析二者之间的相关性;采用ELISA测定血清中IL-6、IL-17、TGF-β1的含量.结果 RSA组RORγt mRNA水平(25.358 ±9.236)显著高于正常对照组(10.611±5.792);Foxp3 mRNA水平(6.851±2.875)显著低于正常对照组(32.218±5.023).RSA患者RORγt和Foxp3 mRNA表达呈显著负相关(r=-0.530,P=0.018).RSA组IL-6、TGF-β1含量分别为(82.7±23.2)、(269.3±29.6) pg/mL,明显低于正常对照组(210.4±40.6)、(497.8±54.6)pg/mL;IL-17含量(172.8±33.9) pg/mL明显高于正常对照组(53.4±10.8) pg/mL.结论 RORγtmRNA表达上调与Foxp3mRNA表达下调所致的Th17和Treg细胞免疫失衡可能是RSA的原因之一;IL-6、IL-17、TGF-β1含量变化的异常在RSA的发生发展中可能起一定作用.     

类风湿性关节炎患者外周血Th17/Treg细胞比率失衡的研究

目的:观察类风湿性关节炎(RA)患者外周血Th17细胞与Foxp3+CD4+CD25+调节性T(Treg)细胞的平衡状态与疾病状态的关系,初步阐明Th17/Treg细胞比率失衡在RA发病机制中的作用和意义。方法:流式细胞术(FCM)检测RA患者和健康人外周血中Th17细胞和Foxp3+CD4+CD25+Treg细胞的比率。结果:活动期RA患者外周血CD3+CD4+T细胞和Th17细胞的比率均明显高于健康对照组(P均0.05);而Foxp3+CD4+CD25+Treg细胞的比率明显低于健康对照组(P0.05)。随疾病活动性的增加,Th17细胞表达增高(P0.05);而Foxp3+CD4+CD25+Treg细胞表达降低,但无统计学意义(P0.05)。结论:RA患者外周血T细胞紊乱以CD4+T细胞的增加为主,Th17细胞比率的增加和Foxp3+CD4+CD25+Treg细胞比率的降低所致的Th17/Treg细胞比率失衡,可能在RA的发生发展中起重要作用。     

溶栓治疗对心肌梗死患者Th17及Treg细胞的影响

目的:观察心肌梗死患者溶栓前后体内Th17、Treg细胞及相关因子含量,从而探讨三者的相关性。方法:提取外周血单个核细胞,流式细胞术检测Th17及Treg含量,ELISA方法检测上清IL-17及TGF-β含量变化,实时定量PCR检测RORγT及Foxp3含量。结果:与对照组相比,实验组患者在溶栓前、后体内Th17细胞,IL-17及RORγT含量均显著升高;Treg细胞、TGF-β及Foxp3含量显著降低(P0.05)。与实验组患者溶栓前相比,溶栓后7 d体内Th17细胞,IL-17及RORγT含量显著降低;Treg细胞、TGF-β及Foxp3含量显著升高(P0.05)。结论:与对照组相比,心肌梗死患者体内中存在Th17及相关因子升高,Treg及相关因子降低,溶栓治疗后可使Th17及相关因子降低,Treg及相关因子升高。     

原发性肾病综合征患儿外周血Th17与CD4~+ CD25~+ Foxp3~+调节性T细胞的水平

目的:观察原发性肾病综合征(PNS)患儿外周血Th17细胞和CD4+ CD25+ Foxp3+调节性T细胞(Treg)及其相关因子的水平和功能,初步探讨其在儿童PNS发病中的作用。方法:PNS患儿分为单纯型肾病(SNS)20例、肾炎性肾病(NNS)15例;同时以20例健康体检儿童作为对照组。采用流式细胞术(FCM)检测外周血单个核细胞(PBMC)中Th17细胞和Treg细胞比例;Real-time PCR法检测PBMC中RORC、IL-23p19和Foxp3 mRNA的表达;ELISA分别检测血清中IL-1β、IL-6、TGF-β1的水平。结果:SNS、NNS组患儿Th17细胞及RORC、IL-23p19 mRNA、IL-1β、IL-6的水平均明显高于正常组(P0.05),且NNS组患儿以上指标明显高于SNS组(P0.05);SNS、NNS组患儿Treg细胞、Foxp3 mRNA和TGF-β1水平的表达明显低于正常组(P0.05),其中两组疾病患儿血清TGF-β1的水平无统计学意义(P0.05),其余指标NNS患儿均明显低于SNS组(P0.05)。结论:Th17/Treg细胞功能失衡可能在儿童PNS的发病中起到重要作用,其失调程度可能与PNS的临床表现、对激素的敏感性、病理类型、及预后有关。     

CD4~+CD25~+Foxp3~+调节性T细胞在子宫内膜癌患者外周血中的表达及意义

探讨在子宫内膜癌患者外周血中CD4+CD25+Foxp3+调节性T细胞的表达情况及意义。采用流式细胞术检测84例术前子宫内膜癌患者及40例子宫肌瘤患者外周血中CD4+CD25+Foxp3+细胞比例及Foxp3平均荧光强度,采用qRT-PCR检测两组患者外周血中Foxp3的mRNA表达情况,同时采用ELISA检测外周血中TGF-β1和IL-17含量。与子宫肌瘤组比较,子宫内膜癌患者外周血中CD4+CD25+Foxp3+Treg细胞的比例虽略有升高但没有统计学意义(P=0.08),而CD4+CD25+细胞内Foxp3的平均荧光强度明显升高(P<0.001)。子宫内膜癌患者外周血中Foxp3的mRNA表达要明显多于子宫肌瘤组(P<0.001)。子宫内膜癌患者外周血中TGF-β1、IL-17的含量要多于子宫肌瘤组。子宫内膜癌患者外周血中的Foxp3+Treg细胞表达增多,这些细胞可能通过增加细胞因子TGF-β和IL-17的分泌从而调节机体对肿瘤细胞免疫反应的方向,最终促进子宫内膜癌的发生和发展。     

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.