冠心病患者血清IL-37水平及其与血清IL-6、IL-10、CRP水平的关系

目的:探讨不同类型冠心病患者血清白介素-37(IL-37)的水平及其与血清白介素-6(IL-6)、白介素-10(IL-10)、C反应蛋白(CRP)水平的关系。方法:选取急性心肌梗死患者20例(AMI组)、不稳定性心绞痛患者26例(UAP组)、稳定性心绞痛患者20例(SAP组)及冠脉造影正常者26例(CON组)为研究对象,采用酶联免疫吸附法(ELISA)测定其血清IL-37、IL-6、IL-10和CRP的水平并分析其相关性。结果:1UAP组、AMI组血清IL-37水平均较对照组(CON组)显著增高(p0.05),而SAP组与CON组比较无明显差异(P0.05)。2冠心病患者的血清IL-37水平与其血清CRP(r=0.3,P0.05)、IL-6(r=0.4,P0.05)水平均存在显著正相关性,与IL-10水平无明显相关(P=0.16)。当排除SAP组患者后,冠心病患者的血清IL-37水平与CRP(r=0.3,P0.05)、IL-6(r=0.5,P0.05)、IL-10(r=0.2,P0.05)水平均显著相关。结论:急性冠脉综合症(ACS)患者的血清IL-37水平显著升高,并与IL-6、IL-10、CRP水平相关,可能参与了ACS发病过程中的炎症反应。     

Recombinant expression of His-tagged saposin B and pH-dependent binding to the lipid coenzyme Q10

The use of coenzyme Q10 (CoQ10) has been increasing rapidly during recent years due to its postulated beneficial properties in human health, providing energy and antioxidant protection. There are no known negative side effects of CoQ10 even at very high levels. Recently, native saposin B (sapB) has been shown to bind CoQ10 and subsequently be excreted. It is thought that this interaction between sapB and CoQ10 could be a mechanism to avoid any possible CoQ10 toxicity. The interaction between sapB and CoQ10 is poorly understood. Here we present an increased fermentative yield of recombinant sapB and demonstrate that recombinant sapB will bind CoQ10 in a pH-dependent manner similar to sapB binding with other lipids. SapB was coated onto an IMAC (immobilized metal affinity chromatography) resin and successfully bound CoQ10 at pH 5.0 with release of the CoQ10 at pH 9.0.     

微生物发酵法生产辅酶Q10研究进展

微生物发酵法是生产辅酶Q10很有前景的方法.本文综述了辅酶Q10产生菌的种类、生物合成机制、辅酶Q10产生菌的改良以及发酵条件优化等方面的研究进展.     

Dosimetric impact of failing to apply correction factors to ion recombination in percentage depth dose measurements and the volume-averaging effect in flattening filter-free beams

PurposeTo examine whether it is essential to apply correction factors for ion recombination (k_S) to percentage depth dose (PDD) measurements and to the volume-averaging effect (k_vol) to ensure accurate absolute dose calibration for flattening filter-free (FFF) beams for the most commonly used ionization chambers.MethodsWe surveyed medical physicists worldwide (n = 159) to identify the five most common ionization chamber combinations used for absolute and relative reference dosimetry of FFF beams. We then assessed the overall absolute dose calibration error for FFF beams of the Artiste Siemens and TrueBeam Varian linear accelerators resulting from failing to apply correction factors k_S in the PDD(10) and the volume-averaging effect (k_vol) to such chamber combinations.ResultsAll the chamber combinations examined—the Farmer PTW 30013 ionization chamber used for absolute dosimetry, and the PTW 31010, PTW 30013, IBA CC04, IBA CC13, and PTW 31021 ionization chambers used for PDD curves measurements—showed non-negligible errors (≥0.5%). The largest error (1.6%) was found for the combination of the Farmer PTW 30013 chamber with the IBA CC13 chamber, which was the most widely used chamber combination in our survey.ConclusionsBased on our findings, we strongly recommend assessing the impact of failing to apply correction factors k_S in the PDD(10) and k_vol prior to using any chamber type for FFF beam reference dosimetry purposes.     

2014年陕西省柯萨奇病毒A10型相关手足口病的流行特征和其VP1区基因特征研究

了解陕西省手足口病(Hand,foot and mouth disease,HFMD)的致病病原体柯萨奇病毒A10型(CV-A10)的流行特征及VP1区基因特征。对2014年收集的HFMD病例标本,通过荧光定量PCR检测确定肠道病毒型别,对CV-A10引起的HFMD流行特征进行描述性分析。使用RD细胞进行病毒分离,RT-PCR扩增CV-A10的VP1区基因片段并进行序列测定,使用Meg Align软件进行核苷酸及氨基酸的同源性分析,并使用MEGA5.0软件构建系统进化树。2014年CV-A10是陕西HFMD病原谱中的第三大病原,占其他肠道病毒的57.71%,13例重症HFMD病例的致病病原体鉴定为CV-A10,占重症病例的9.03%。CV-A10感染HFMD病例以≤3岁年龄组儿童为主(83.07%),男女性别比为1.15∶1。发病时间主要集中在4～7月。实验室分离出101株CV-A10,覆盖全省10市(区)。完成测序的18株CV-A10核苷酸和氨基酸同源性分别为94.0%～100.0%和97.3%～100.0%,与A型原型株的核苷酸和氨基酸同源性分别为76.2%～77.5%和91.9%～93.0%,与近年来河北、湖南和河南地区流行株具有较高的同源性。系统进化显示陕西CV-A10分离株属于C基因型。CV-A10是2014年陕西HFMD的优势病原,能引起重症HFMD,本次分离到的CV-A10毒株均属于C基因型。     

黄色隐球酵母生产辅酶Q10发酵条件的优化

从黄色隐球酵母L3302提取辅酶Q10,经过氮源、碳源、初始pH、发酵温度等的研究分析,得到最佳的发酵条件。通过最优化实验确定培养基:蔗糖和葡萄糖各1.25g/L;酵母膏和玉米浆各0.3g/L;pH值6.5,温度28℃,接种量5%,装液量为50mL/500mL;生长因子以蛋白水解液为优。按此发酵条件上罐发酵,得到菌体生长量为12.8g/L发酵液,辅酶Q10的产量为1.82mg/100mL发酵液。     

EB病毒白介素-10基因在广东地区人群中的变异特征及与鼻咽癌的相关性

EB病毒（Epstein-Barr virus,EBV）基因变异与EBV相关疾病发生的关系尚无明确结论,本研究旨在了解广东地区人群中EBV白介素-10基因（viral interleukin-10,vIL-10）基因变异特征,探讨其变异与鼻咽癌（nasopharyngeal carcinoma,NPC）发生的关系。采用PCR和DNA测序检测NPC组织和健康成年人咽漱液中vIL-10序列。50例NPC和70例健康人群完成序列测定,其中42例（84.0%）NPC和60例（85.7%）健康人群与EBV标准株B95-8氨基酸序列一致,被分类为B95-8型;剩余8例（16.0%）NPC和10例（14.3%）健康人群在信号肽区域出现氨基酸突变,被分类为SPM变异型。2种型别及4处共有突变在2种人群中的分布无统计学意义（P>0.05）。结果表明,广东地区NPC和健康人群中EBV vIL-10高度保守,均以B95-8型为主,而SPM变异型的地域性分布及与鼻咽癌的确切关系有待进一步研究。     

Immunoregulation through IL-10 gene expression and the fate of cytotoxic T lymphocyte-mediated tumor immunotherapy

Gene analysis of tumor associated antigens revealed that tumor antigens are all normal gene product. Inducing tumor reactive cytotoxic T lymphocytes (CT) in the patients is same as inducing autoimmunity in the patients. Immunosuppressive cytokine interleukin-10 (IL-10) plays an important role in maintaining homeostasis or tolerance. To break the tumor tolerance, blocking and IL-10 secretion or intervention in the pathways of IL-10 gene activation is indeed important.     

表皮生长因子对转化的小鼠成纤维细胞DNA拓扑异构酶活性的影响

我们用表皮生长因子对用氚标记的脱氧胸苷转化的C_3H_(10)T1/2 CL8细胞进行刺激,研究了表皮生长因子对该细胞胞浆及胞核内DNA拓扑异构酶活性的影响,结果发现,表皮生长因子可以使转化的小鼠成纤维细胞胞浆和胞核内的DNA拓扑异构酶活性增加。     

The Establishment of Infectious Clone and Single Round Infectious Particles for Coxsackievirus A10

Coxsackievirus A10(CVA10) is one of the major etiological agents of hand, foot, and mouth disease. There are no vaccine and antiviral drugs for controlling CVA10 infection. Reverse genetic tools for CVA10 will benefit its mechanistic study and development of vaccines and antivirals. Here, two infectious clones for the prototype and a Myc-tagged CVA10 were constructed. Viable CVA10 viruses were harvested by transfecting the viral m RNA into human rhabdomyosarcoma(RD)cells. Rescued CVA10 was further confirmed by next generation sequencing and characterized experimentally. We also constructed the vectors for CVA10 subgenomic replicon with luciferase reporter and viral capsid with EGFP reporter,respectively. Co-transfection of the viral replicon RNA and capsid expresser in human embryonic kidney 293 T(HEK293 T)cells led to the production of single round infectious particles(SRIPs). Based on CVA10 replicon RNA, SRIPs with either the enterovirus A71(EVA71) capsid or the CVA10 capsid were generated. Infection by EVA71 SRIPs required SCARB2,while CVA10 SRIPs did not. Finally, we showed great improvement of the replicon activity and SRIPs production by insertion of a cis-active hammerhead ribozyme(HHRib) before the 50-untranslated region(UTR). In summary, reverse genetic tools for prototype strain of CVA10, including both the infectious clone and the SRIPs system, were successfully established. These tools will facilitate the basic and translational study of CVA10.     

过表达白细胞介素10的间充质干细胞对炎症性肠病的治疗作用

炎症性肠病(inflammatory bowel disease,IBD)是一种以T细胞浸润至结肠为特征的难治性炎性自身免疫疾病。间充质干细胞 (mesenchymal stem cells,MSCs) 具有免疫抑制能力,在IBD的治疗中具有一定的潜力。但是由于MSCs在体内的免疫调节能力不稳定,所以其治疗效果会受到影响。本研究构建了过表达白细胞介素10(interleukin 10,IL-10)的工程化MSCs,并对其在IBD小鼠模型中的治疗潜力进行评估。MSCs经编码IL-10的慢病毒(lentivirus,LV)转染后,其表型和细胞增殖率均不发生变化。免疫细胞和 MSCs体外共培养的结果表明,与未修饰的MSCs相比,同过表达IL-10的MSCs共培养的免疫细胞中辅助T细胞1(T helper 1 cells,Th1)和辅助T细胞17(T helper 17 cells,Th17)数量显著性降低(P<0.05),同过表达IL-10的MSCs共培养的巨噬细胞细胞培养上清液,TNF-α含量显著性降低(P<0.0001)。右旋糖酐硫酸钠(dextran sodium sulfate,DSS)诱导IBD小鼠模型中,尾静脉注射过表达IL-10的MSCs与注射未修饰MSCs相比,过表达IL-10的MSCs具有更好的治疗效果,结肠长度、疾病活动指数(disease activity index,DAI)和结肠炎性细胞因子表达共同证明这一差异。实验结果均具有统计学差异(P<0.05)。总体而言,经LV转染过表达IL-10的MSCs可能是IBD的一种有希望的替代治疗选择。     

USP10 exacerbates neointima formation by stabilizing Skp2 protein in vascular smooth muscle cells

The underlying mechanism of neointima formation remains unclear. Ubiquitin-specific peptidase 10 (USP10) is a deubiquitinase that plays a major role in cancer development and progression. However, the function of USP10 in arterial restenosis is unknown. Herein, USP10 expression was detected in mouse arteries and increased after carotid ligation. The inhibition of USP10 exhibited thinner neointima in the model of mouse carotid ligation. In vitro data showed that USP10 deficiency reduced proliferation and migration of rat thoracic aorta smooth muscle cells (A7r5) and human aortic smooth muscle cells (HASMCs). Mechanically, USP10 can bind to Skp2 and stabilize its protein level by removing polyubiquitin on Skp2 in the cytoplasm. The overexpression of Skp2 abrogated cell cycle arrest induced by USP10 inhibition. Overall, the current study demonstrated that USP10 is involved in vascular remodeling by directly promoting VSMC proliferation and migration via stabilization of Skp2 protein expression.     

Bilobalide regulates soluble amyloid precursor protein release via phosphatidyl inositol 3 kinase-dependent pathway

Bilobalide (BB) is a sesquiterpenoid extracted from Ginkgo biloba leaves. An increasing number of studies have demonstrated its neuroprotective effects. The neuroprotective mechanisms may be associated with modulation of intracellular signaling cascades such as the phosphatidyl inositol 3-kinase (PI3K) pathway. Using differentiated SH-SY5Y cells, this study investigated whether BB modulation of intracellular signaling pathways, such as the protein kinase C (PKC) and PI3K pathways, contributes to amyloid precursor protein (APP) metabolism, a key event in the pathogenesis of Alzheimer’s disease (AD). We demonstrated in this study that BB enhanced the secretion of α-secretase-cleaved soluble amyloid precursor protein (sAPPα, a by-product of non-amyloidogenic processing of APP) and decreased the β amyloid protein (Aβ, a by-product of amyloidogenic processing of APP) via PI3K-dependent pathway. The PI3K pathway mediated the rapid effect of BB on APP processing possibly via regulation of intracellular APP trafficking. After longer time BB incubation (12 h), this effect was reinforced by PI3K pathway-mediated up-regulation of disintegrin and metalloproteinase domain-containing protein 10 (ADAM10, an α-secretase candidate). Given the strong association between APP metabolism and AD pathogenesis, the ability of BB to regulate APP processing suggests its potential use in AD prevention.