慢性乙型肝炎患者外周血SOCS3 mRNA的表达及其与Th17的关系

 目的 研究慢性乙型肝炎(CHB)患者外周血辅助性T细胞17(Th17)以及细胞因子信号转导抑制因子3(SOCS3) mRNA的表达状况,探索CHB患者SOCS3 mRNA表达与Th17的关系。方法 选取2021年2—8月于某院门诊就诊的30例CHB患者为研究对象,并选取同期正常体检者15例为对照组。采用流式细胞术(FCM)检测外周血Th17细胞频数,酶联免疫吸附试验(ELISA)检测血清细胞因子IL-17A和IL-23表达水平,实时荧光定量逆转录聚合酶链反应(qRT-PCR)法测定外周血SOCS3、维甲酸相关孤儿核受体γt (RORγt) mRNA表达水平,并比较两组患者的检测结果。结果 CHB患者外周血Th17细胞频数及其效应分子IL-17A、IL-23表达水平高于对照组(均P＜0.05),Th17细胞频数、IL-17A与HBV DNA水平之间存在正相关(r值分别为0.570、0.563,均P＜0.005)。CHB患者外周血SOCS3、RORγt mRNA表达水平高于对照组(均P＜0.05),两者与HBV DNA水平正相关(r值分别为0.662、0.561,均P＜0.05)。CHB患者SOCS3 mRNA与RORγt mRNA、Th17细胞频数、IL-17A之间存在正相关(r值分别为0.552、0.626、0.826,均P＜0.05)。结论 CHB患者外周血SOCS3 mRNA的异常高表达,可能通过调节RORγt mRNA的表达来影响Th17的分化及其效应分子的分泌。     

人食管癌相关基因cDNA片段的克隆、筛选与初步鉴定

目的 :筛选和鉴定人原发性食管癌组织的相关基因 ,揭示食管癌的癌变机理。方法 :用高效、灵敏的荧光mRNA差异显示技术 ,以食管癌及相应的正常食管粘膜组织为对照 ,通过对其基因表达的比较 ,找出差异条带 ,利用ReverseNorthernDotBlot、DNA序列分析和NorthernBlot,并结合mRNA原位杂交技术对被筛片段进行鉴定。结果 :①在实验中分离、鉴定了 74条差异片段 ,其中包括正常组织表达而癌组织中不表达的差异片段 5 4个 ,癌组织中表达而正常组织不表达的差异片段 2 0个。②其中 1个片段C5 7(337bp)在GenBank数据库中没有发现同源的已知基因或片段 ,推测其可能为食管癌相关基因。③经NorthernBlot检测C5 7在癌组织中有表达信号。④原位杂交技术检测C5 7在癌组织中具有较高的阳性表达率 (74% )。结论 :食管癌组织中C5 7可能是新的食管癌相关的候选癌基因 ,它与食管癌的发生发展可能有关     

Discovery and Validation of a Urinary Exosome mRNA Signature for the Diagnosis of Human Kidney Transplant Rejection

BackgroundDeveloping a noninvasive clinical test to accurately diagnose kidney allograft rejection is critical to improve allograft outcomes. Urinary exosomes, tiny vesicles released into the urine that carry parent cells’ proteins and nucleic acids, reflect the biologic function of the parent cells within the kidney, including immune cells. Their stability in urine makes them a potentially powerful tool for liquid biopsy and a noninvasive diagnostic biomarker for kidney-transplant rejection.MethodsUsing 192 of 220 urine samples with matched biopsy samples from 175 patients who underwent a clinically indicated kidney-transplant biopsy, we isolated urinary exosomal mRNAs and developed rejection signatures on the basis of differential gene expression. We used crossvalidation to assess the performance of the signatures on multiple data subsets.ResultsAn exosomal mRNA signature discriminated between biopsy samples from patients with all-cause rejection and those with no rejection, yielding an area under the curve (AUC) of 0.93 (95% CI, 0.87 to 0.98), which is significantly better than the current standard of care (increase in eGFR AUC of 0.57; 95% CI, 0.49 to 0.65). The exosome-based signature’s negative predictive value was 93.3% and its positive predictive value was 86.2%. Using the same approach, we identified an additional gene signature that discriminated patients with T cell–mediated rejection from those with antibody-mediated rejection (with an AUC of 0.87; 95% CI, 0.76 to 0.97). This signature’s negative predictive value was 90.6% and its positive predictive value was 77.8%.ConclusionsOur findings show that mRNA signatures derived from urinary exosomes represent a powerful and noninvasive tool to screen for kidney allograft rejection. This finding has the potential to assist clinicians in therapeutic decision making.     

Second dose of the BNT162b2 mRNA vaccine: Value of timely administration but questionable necessity among the seropositive

This study monitored titers of neutralizing IgG against the receptor-binding domain of the SARS-CoV-2 S1 subunit 14 days post-injection of each dose of the BNT162b2 mRNA Covid-19 vaccine in 401 Greek healthcare workers aged 20–67. After the first dose, titers varied upon age and history of infection, being lower in the 50+ age group and significantly higher among the seropositive. After the second dose, immunogenicity was significantly boosted in the age 50+ and SARS-CoV-2-naïve individuals, indicating the effectuality of its timely administration, yet questioning its value among the seropositive.     

The neurostructural and neurocognitive effects of physical activity: A potential benefit to promote eating disorder recovery

Accumulating evidence suggests that supervised and adapted physical activity provides cognitive benefits for individuals with eating disorders (EDs). The mechanisms underlying the benefits of physical activity are poorly understood. Addressing this knowledge gap may inform the appropriate integration of structured physical activity into eating disorders treatment and recovery. We draw attention to recent findings in the study of the impact of physical activity on the brain, and we describe the neurostructural and neurocognitive changes associated with physical activity observed in various clinical and nonclinical populations. Considering the identified impairment in brain volume- and/or neurocognitive function in various EDs, we propose that positive effects of physical activity may play a meaningful role in successful ED treatment. Accordingly, we outline research steps for closing the knowledge gap on how physical activity may aid in ED recovery, and emphasize the need to combine measures of cognitive and behavioral responses to physical activity, with technology capable of measuring changes in brain structure and/or function.     

运动强度对幼龄大鼠海马VEGF及其受体Flk-1 mRNA表达的影响

为了研究不同强度跑台运动对5周龄大鼠海马表达血管内皮生长因子(vascular endothelial growth factor,VEGF)和胎肝激酶-1(fetal liver kinase-1,Flk-1)受体mRNA水平的影响,5周龄雄性SD大鼠被随机分为对照组和小强度、中等强度、大强度3个跑台运动组,并通过定量real-time PCR技术,观察不同强度跑台运动1周后大鼠海马内VEGF和Flk-1mRNA表达的影响。采用3-磷酸甘油醛脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)作为管家基因内参,以样品中所测得的VEGF和Flk-1基因拷贝数除以该样品中内参基因拷贝数进行校正后,各组动物海马内VEGF mRNA相对水平分别是:对照组为2740±2229,小强度运动组为3599±2386(P<0.05),中等强度运动组为1654±1136,大强度运动组为1572±1071。各组动物海马内Flk-1mR-NA相对水平分别是:对照组为147±47,小强度运动组为281±182(P<0.01),中等强度运动组为208±100;大强度运动组为54±31。以上结果提示,大鼠海马内VEGF和Flk-1mRNA的表达受运动的调节,且与运动强度有关。小强度跑台运动可对VEGF和Flk-1mRNA的表达产生促进作用,而大强度运动则不影响VEGF和Flk-1mRNA的表达。     

用非同位素原位杂交组化在光镜和电镜水平观察前阿黑皮素mRNA在垂体的分布

用地高辛标记反意前阿黑皮素(pro-opiomelanocortin,POMC)cRNA探针原位杂交组化在光镜和电镜水平观察了POMC mRNA在大鼠垂体的分布,并比较了碱性磷酸酶(AKP)显色系统和辣根过氧化酶(HRP)显色系统在光镜水平上的敏感性.结果:POMC mRNA广泛地分布于垂体的中间叶和前叶.中间叶全部细胞均为POMC mRNA阳性.前叶中除前叶的腹侧缘和前叶与中间叶交界处阳性细胞较少外,都有较多的POMC mRNA阳性细胞分布.AKP显色系统比HRP显色系统敏感.在电镜水平,POMC mRNA主要分布于粗面内质网,少数分泌颗粒可能呈阳性反应,胞核未见阳性反应沉淀.文内还就阳性分泌颗粒在调节细胞合成功能方面可能起的作用进行了讨论.     

大鼠延髓内含前-原脑啡肽mRNA、甲硫氨酸-脑啡肽和亮氨酸-脑啡肽神经元的分布

应用原位杂交和免疫组织化学方法,对成年大鼠延髓内含前-原脑啡肽(PPE)mRNA和甲硫氨酸-脑啡肽(M-ENK)与亮氨酸-脑啡肽(L-ENK)免疫反应神经元进行观察。结果表明:含PPEmRNA的神经元胞体,多数分布在孤束核、腹外侧区以及两者之间的网状结构等形成的一条从背内侧到腹外侧区的弧形带内。M-ENK与L-ENK样免疫反应阳性结构(神经元胞体、纤维和终末)也主要密集分布在该带内。本结果对ENK(M-ENK和L-ENK)参与延髓内脏功能活动的调控过程,提供了进一步的形态学证据。     

大鼠卵泡发育过程中基质金属蛋白酶-2，-9的作用

目的：探讨基质金属蛋白酶-2,-9（MMP-2,-9）在大鼠卵泡发育的组织重建中的作用。方法：采用原位杂方法检测动情周期大鼠卵泡中MMP-2,-9mRNAs的基因表达变化,并通过密度扫描方法对原位杂交结果进行半定量分析。结果：在动情周期中,各情期卵巢中卵泡的颗粒细胞和发育良好的卵泡的膜细胞中均可检测到MMP-2m-RNA的杂交信号;MMP-9mRNA的表达始于已发育良好的窦前卵泡的膜细胞,此后在小窦状卵泡、窦状卵泡及排卵前卵泡的膜细胞中均检测到MMP-9mRNA的杂交信号。结论：MMP-2,-9可能通过其基因的表达变化参与卵泡的发育过程。