Th17/Treg细胞失衡与子宫颈癌

Th17细胞是近年发现的一种新型CD4^＋效应性T细胞,以其特异性的分泌IL-17而命名。介导免疫耐受的Treg细胞和介导炎症反应的Th17细胞间功能和分化过程相互对抗,在正常状态下,两者保持平衡,但机体发生功能异常时常表现出Treg/Th17失衡,引起炎性反应、自身免疫性疾病、移植物抗宿主病等的发生和发展,并决定疾病的转归和预后,在肿瘤免疫中亦发挥了重要作用。该文就Treg/Th17失衡在肿瘤,尤其是子宫颈癌发生发展中的作用进行综述。     

IL-17/Th17 targeting: on the road to prevent chronic destructive arthritis?

Interleukin-17A (IL-17A) contributes to the pathogenesis of arthritis. Data from experimental arthritis indicate IL-17 receptor signaling as a critical pathway in turning an acute synovitis into a chronic destructive arthritis. The identification of six IL-17 family members (IL-17A-F) may extend the role of this novel cytokine family in the pathogenesis of chronic destructive joint inflammation. Whether the successful anti-IL-17A cytokine therapy in murine arthritis can be effectively translated to human arthritis need to be tested in clinical trials in humans. Interestingly, IL-17A and IL-17F are secreted by the novel T helper subset named Th17. This novel pathogenic T cell population induces autoimmune inflammation in mice and is far more efficient at inducing Th1-mediated autoimmune inflammation in mice than classical Th1 cells (IFN-gamma). In addition to IL-17A and IL-17F, Th17 cells are characterized by expression of IL-6, TNF, GM-CSF, IL-21, IL-22 and IL-26. Th17 cells have been established as a separate lineage of T helper cells in mice distinct from conventional Th1 and Th2 cells. Whether this also applies to human Th17 and whether RA is a Th1 or a Th17 mediated disease is still not clear. This review summarizes the findings about the role of IL-17 in arthritis and discusses the impact of the discovery of the novel Th17 cells for arthritis. Further studies are needed to unravel the role of Th17 cells and the interplay of IL-17 and other Th17 cytokines in the pathogenesis of arthritis and whether regulating Th17 cell activity will have additional value compared to neutralizing IL-17A activity alone. This might help to reach the ultimate goal not only to treat RA patients but to prevent the development of this crippling disease.     

Identifying a novel mutation of CYP17A1 gene from five Chinese 17α-hydroxylase/17, 20-lyase deficiency patients

Mutations of CYP17A1 gene could cause complete or partial, combined or isolated 17α-hydroxylase/17,20-lyase enzyme deficiencies (17OHD). We intended to investigate the CYP17A1 mutation in five unrelated patients and analyze its possible influence on phenotype of an atypical 17OHD patient presented with micropenis, hypertension and intermittent hypokalemia. Steroid hormones were assayed in these patients. A novel missense mutation (c.1169C>G, p. Thr390Arg) located in exon 7 was detected in one of the patients. Homozygous c. 985_987delinsAA, p. Tyr329fs mutation was found in two patients, while compound heterozygous mutations (c. 985_987delinsAA, p. Tyr329fs/c. 932–939 del, p. Val311fs and c. 287G>A, p. Arg96Gln/c. 985_987delinsAA, p. Tyr329fs) were found in two other patients, respectively. Then, steric model analysis of CYP17A1 showed that the novel mutation T390R changed the local structure as well as the electrostatic potential of the nearby beta sheet. Finally, site-directed mutagenesis and in vitro expression were used to analyze the activity of novel mutant CYP17A1. It indicated the T390R mutant retained part of enzyme activity, which was consistent to the clinical features. In conclusion, we identified a novel missense mutation of CYP17A1 gene from a patient with micropenis, hypertension and intermittent hypokalemia, which varied from other four patients. It also expanded our understanding of genotype–phenotype correlation of the disease.     

Microbial transformation of 17α-cyanomethyl-17-hydroxy-4,9-estradien-3-one (STS 557) and 17α-cyanomethyl-19-nortestosterone by Mycobacterium smegmatis

Microbial transformation of the new progestagen STS 557 (17α-cyanomethyl-17-hydroxy-4,9-estradien-3-one) by $\text{Mycobacterium smegmatis}$ yielded predominantly ring A-aromatized compounds: 17α-cyanomethyl-1,3,5(10),9(11)-estratetraene-3, 17-diol, 17α-cyanomethyl-1,3,5(10)-estratriene-3, 17-diol and the corresponding 3-methyl ethers. The analogous compound without the 9(10) double bond, 17α-cyanomethyl-19-nortestosterone, was transformed mainly to 5α-hydrogenated metabolites: 17α-cyanomethyl-17-hydroxy-5α-estran-3-one, 17α-cyanomethyl-17-hydroxy-5α-1-estren-3-one, 17α-cyanomethyl-5α-estrane-3α, 17-diol, and 17α-cyanomethyl-5α-estrane-3β, 17-diol. From these results, it is concluded that 4,9-dien-3-oxo compounds are not substrates for enzymatic 5α-hydrogenation.     

13/17染色体易位纯合子猪DNA指纹分析

13/17染色体易位纯合子猪是孙金海等于1991年通过易位杂合子互交在国内外首次获得的一种新遗传类型家猪[2n=36,XY或XXrob(13;17)]。本研究用噬菌体M13mp18单链小卫星DNA作探针对该种新遗传类型猪、13/17染色体易位杂合子猪及正常核型家猪进行了DNA指纹分析。采用限制性内切酶HinfⅠ和HaeⅢ均成功地获得高分辨的、具有个体特异性的DNA指纹图谱。研究结果表明,不同内     

Molecular basis of 17α-hydroxylase/17,20-lyase deficiency

17α-Hydroxylase deficiency is characterized by a defect in either or both of 17α-hydroxylase and 17,20-lyase activities, based on the fact that a single polypeptide P450c17 can catalyze both reactions. The clinical manifestations of 17α-hydroxylase/17,20-lyase deficiency seem to be more heterogeneous than expected, varying from the classical type to less symptomatic forms as also observed in 21-hydroxylase deficiency. We have sequenced all eight exons of the CYP17 (P450c17) gene in DNA from several patients, reconstructed the mutations in a human P450c17 cDNA and expressed the mutant P450c17 in COS 1 cells to characterize the kinetic properties of 17α-hydroxylase and 17,20-lyase activities. The molecular bases of cases clinically reported as 17α-hydroxylase deficiency have turned out to be complete or partial combined deficiencies of 17α-hydroxylase/17,20-lyase. The elucidation of the molecular basis generally explains the patient's clinical profiles including the sexual phenotype of the external genitalia. In one case clinically reported as isolated 17,20-lyase deficiency, the molecular basis was found to be partial combined deficiency of both activities, somewhat discordant with the patient's clinical profile. Based on the results obtained so far we can predict that those 17α-hydroxylase deficient individuals having a homozygous stop codon in the CYP17 gene positioned at the amino terminal side of the P450c17 heme-binding cysteine (442) will all have the same phenotype. However those individuals having homozygous missense mutations or those who are compound heterozygotes having a missense mutation in at least one CYP17 allele will display their own unique phenotype which clinically will be subtly different from all others.     

ADAM17通过EGFR-PI3K/Akt/p27通路调控前列腺癌细胞增殖

ADAM17是金属蛋白酶家族(ADAMs)成员之一,研究发现ADAM17可以通过水解细胞表面蛋白的胞外结构域导致肿瘤细胞的增殖和转移.本课题前期研究结果显示,与LNCap细胞相比,ADAM17在DU145细胞中高表达,且与细胞增殖相关.为了研究ADAM17与前列腺癌细胞增殖相关基因p27表达的关系及调控机制,我们采用RNAi技术下调ADAM17的表达,加入PMA(一种ADAM17的激活剂)上调ADAM17的表达,通过细胞计数和CCK-8方法检测细胞增殖,RT-PCR检测p27mRNA的表达,Western印迹检测ADAM17的表达;进一步阻断EGFR和PI3K/Akt信号转导,RT-PCR方法检测p27mRNA的表达,Western印迹检测ADAM17、EGFR、pEGFR、Akt和pAkt的表达.结果显示ADAM17的表达与前列腺癌细胞的增殖呈正相关(P0.05);p27mRNA的表达与ADAM17的表达呈负相关(P0.05);分别阻断EGFR和PI3K/Akt信号转导通路,同时使ADAM17表达增加,与对照组(单独PMA处理组)相比,p27mRNA的表达均增加(P0.05).提示ADAM17调控前列腺癌细胞增殖相关基因p27表达是通过EGFR-PI3K/Akt信号通路实现的.     

Treg/Th17平衡失调在复发性流产中的作用

正常妊娠被认为是一种成功的半同种移植,有赖于母胎界面生理性抑制反应的增强,称为免疫耐受。孕妇的免疫系统失调可以导致生殖失败,同种免疫的异常很有可能是复发性流产的发病机理。随着对免疫耐受机制研究的不断深入,母胎免疫耐受的研究从Th1/Th2细胞平衡失调进一步发展到对Treg/Th17细胞平衡失调的研究,该文综述了Treg/Th17细胞平衡失调在原因不明复发性流产中的作用。     

Th17/Treg 细胞在银屑病的发病机制研究进展

Th17细胞和Treg细胞是CD4+T细胞在不同细胞因子环境中分化出的新亚群,发挥不同的生物学效应,使机体的免疫系统处于平衡状态.Th17/Treg细胞失衡可引起一系列自身免疫性疾病.银屑病是与遗传、免疫异常有关的皮肤炎症性疾病,其发病机制尚不清楚.越来越多的研究发现,Th17细胞增多和Treg细胞减少及其分泌的细胞因子在银屑病的发病中有着重要作用.本文围绕这一机制综述了近年来有关Th17细胞、Treg细胞在银屑病发病机制中作用的研究,帮助我们更深入地了解银屑病的发病机制并为今后临床诊断和治疗提供依据.     

IL-23/Th17轴在变应性哮喘中的研究进展

变应性哮喘是一种由辅助性T细胞（T helper cell,Th cell）调节的慢性炎症性疾病。Th1/Th2的失衡一直被认为是变应性哮喘的发病机制,Th2细胞及其分泌的细胞因子白介素4（interleukin 4,IL-4）、IL-5以及IL-13在变应性哮喘特异性症状的发病中发挥重要作用。最近研究发现Th17细胞及其分泌的IL-17参与变应性哮喘的发展过程,IL-23在Th17细胞维持生存和功能成熟中发挥重要作用,并参与抗原诱导的气道炎症反应。该文对目前IL-23/Th17轴在变应性气道炎症反应中的研究进展作一综述。     

Th17/Treg失衡与肝脏免疫病理反应的研究进展

Th17细胞及Th17/Treg失衡在炎症反应、组织损伤及纤维化形成中发挥了重要作用,与多种疾病的发生发展密切相关。前炎性细胞因子可诱导T细胞分化为Th17,使Th17/Treg失衡,导致IL-17、IL-6、趋化因子等促炎性细胞因子大量分泌并有效介导中性粒细胞动员与兴奋,使得机体产生炎症反应与免疫病理反应。就Th17/Treg细胞及其失衡在肝脏免疫病理反应中的研究进展进行了综述。     

高粱SbSBP17基因的克隆及表达分析

SBP box(Squamosa promoter Binding Protein box)蛋白是绿色植物中特有的一类转录因子,其功能涉及植物叶片发育、胚胎发生、间隔期长度、营养到生殖生长的更替、育性维持等生长发育的重要过程。该研究以高粱材料BTx623花序总RNA为模板进行RT PCR,克隆了高粱SbSBP17基因,并对其进行了系统进化及半定量PCR表达分析,为进一步研究SbSBP17基因的生物学功能奠定基础。结果表明：SbSBP17含有2个内含子和3个外显子,编码1个444氨基酸的蛋白质,在其195~269 aa区域含有一个典型的SBP box结构域;系统进化分析表明,18个SBP17蛋白可分为三组,第Ⅰ和Ⅱ组只有单子叶植物基因,而第Ⅲ组只有双子叶植物基因,SbSBP17与玉米PWZ17260.1亲缘关系最近;启动子顺式作用元件分析显示,SbSBP17启动子含有细胞周期调控元件MSA like、分生组织发育调控相关元件CAT box和组织特异性表达元件RY element等。基因表达分析显示,SbSBP17是花序特异性表达基因,随着花序发育(长度增加),SbSBP17基因表达量逐渐增高,当花序长至10 cm时基因表达量达到峰值,约是花序长度最小点(≤2 cm)时的40倍,随后表达量逐渐降低,并于花序长度20 cm时显著降低50%以上。研究推测,随着高粱花序进一步发育,SbSBP17基因的表达量可能逐渐降低,最终消失。     

白介素17

白介素１７黄仕和,秦椿华（卫生部武汉生物制品研究所,武汉４３００６０）（同济医科大学工业毒理研究室,武汉４３００３０）关键词白介素１７新近,美国Ｉｍｍｕｎｅｘ公司的尧政兵等（１９９５）从激活的人Ｔ细胞里鉴定了一种新型白介素—ＩＬ－１７（ｈＩＬ－１７）...     

百合LbKCS5和LbKCS17基因的克隆与表达分析

β 酮脂酰辅酶A合成酶(KCS)是超长饱和脂肪酸链(VLCFAs)生物合成中的限速酶,该研究以百合(Lilium brownii var. viridulum Baker.)‘黄天霸’cDNA为模版, 采用RT PCR方法克隆了LbKCS5和LbKCS17基因的CDS序列,并进行序列分析,采用qRT PCR方法分析其基因表达以及胁迫诱导特征。结果表明：(1)LbKCS5和LbKCS17分别包含689和1 238 bp完全阅读框(ORF), 编码186和291个氨基酸;进化分析显示, LbKCS5与油棕KCS5、LbKCS17与莲KCS17序列相似性最高,分别为82.61%和77.62%。(2)基因表达分析结果显示, LbKCS5和LbKCS17 在百合营养器官中均有表达,二者在叶中表达量最高,在根中最少;在花器官中,LbKCS5的表达量在雄蕊中最高,LbKCS17在花药中最高,在花瓣中二者的表达量最低;在花蕾生长发育过程中,花瓣中2个基因的表达均先升高后下降,并于黄蕾期达到峰值;叶中LbKCS5表达量在黄蕾期达到最高,而LbKCS17在叶中的表达整体较低。(3)失水胁迫能提高这2个基因的转录表达,低温诱导LbKCS5表达量的增加,但却降低了LbKCS17的表达。研究表明,LbKCS5和LbKCS17在百合不同器官中均有表达,且均响应失水和低温胁迫,这为研究百合耐失水和低温胁迫以及新品种选育提供了理论支持。     

17-Hydroxyprogesterone metabolism in the monkey fetal adrenal: C17–20Lyase and 21-hydroxylase activities

C^17–20Lyase and 21-hydroxylase activities were measured during late gestation In the rhesus monkey ($\text{Macaca mulatta}$) fetal adrenal. Activities were assessed in 10,000 × g supernatants with 17-hydroxyprogesterone and NADPH as substrates. Although conversion of [¹⁴C]17-hydroxyprogesterone to [¹⁴C]androstenedione was noted, activity was often nonlinear and far less than the rate of hydroxylation which together prevented an accurate estimation of lyase rate, K_m and V_max. 21-Hydroxylase activity was characterized; the mean reaction rate was 1.6 × 10^?3 μmoles NADPH oxidized/min. × mg^?1 protein with an apparent K_m of 3.6 × 10^?7 M and a V_max of 2.2 × 10^?3 μmoles/min. × mg^?1 protein. These values were similar to data obtained In adrenals from adult monkeys. A relatively high level of hydroxylase activity in the fetal gland might lead to an Inadequate supply of precursors for the synthesis of dehydroepiandrosterone sulfate (DHEAS) in the adrenal if it also contained 3β-hydroxysteroid dehydrogenase (3β-hsdh). However, the fact that the fetal adrenal reportedly is deficient in 3β-hsdh may serve to protect both DHEAS and corticoid synthesis.     

13/17罗伯逊易位猪POU1F1基因多态性

采用外周血淋巴细胞培养制备染色体标本, 对13/17罗伯逊易位猪的3种杂交组合的394头后代进行核型分析, 出现3种核型猪：13/17易位纯合子猪[2n=36, XY或XX, rob(13;17)]、13/17易位杂合子猪[2n=37, XY或XX, rob(13;17)]和正常核型猪[2n=38, XY或XX]。应用PCR-RFLP技术在POU1F1基因的1 746 bp扩增片段中检测到1个RsaⅠ限制性内切酶的多态位点。应用PCR-SSCP技术检测POU1F1基因第4外显子, 在3种杂交后代群中均未检测到突变。遗传多态性分析结果表明：RsaⅠ酶切多态位点的突变在3种杂交后代群中A等位基因和AA基因型频率占优势, 在3种核型群体中也是A等位基因和AA基因型频率占优势, 其中AB基因型频率在易位杂合型群体中较高。各杂交后代群均未达到Hardy-Weinberg平衡, 不同核型群亦处于非平衡状态。13/17易位杂合子猪×13/17易位杂合子猪和13/17易位杂合子猪×约克两杂交后代群的PIC表现为中度多态, 而13/17易位杂合子猪×皮杜杂交后代群表现为低度多态; 易位杂合型群体表现为中度多态, 正常核型和易位纯合型群体表现低度多态性。     

IL-23/IL-17轴在脓毒症中的表达及意义

目的:探讨IL-23/IL-17轴在脓毒症患者中的表达及意义.方法:符合诊断标准的脓毒症患者40例,以28天预后为终点,将患者分为存活组(n=21)和病死组(n=19),分析各组病人的急性生理和慢性健康评分(APACHE)Ⅱ和序贯器官衰竭估计(SOFA)评分,同时在入ICU第1天采取外周静脉血做IL-23和IL-17检测,并对病死率和IL-23、IL-17、APACHEⅡ、SOFA做相关性分析.结果:与存活组比较,病死组患者拥有较高的APACHEⅡ和SOFA评分(P<0.01),且外周血的IL-23和IL-17蛋白含量均明显升高(P<0.05).APACHEⅡ和SOFA评分、IL-17和IL-23含量与28天预后有明显的相关性(P<0.05).结论:脓毒症Th17细胞分泌的IL-23/IL-17增加,加重患者病情,在脓毒症发病机制中可能扮演重要角色.     

核苷对ROS17/2.8细胞株腺苷酸环化酶活性的影响

 大鼠成骨肉瘤细胞株(ROS17/2.8)系甲状旁腺素(PTH)的靶细胞。当该细胞质膜上的PTH受体与PTH结合后,可激发腺苷酸环化酶(AC)的活性。腺苷及四种不同的核苷酸(AMP、GMP、UMP、CMP)单独对AC无明显效应,但却可抑制PTH对AC的刺激作用。而鸟苷或木糖腺苷则可显著增强PTH对AC的刺激作用。提示核苷的不同代谢物在代谢调节中的多样化作用。     

荧光假单胞菌CLW17菌株pqqE和GDH基因的功能

[背景]磷是植物生长所必需的大量元素,但绝大多数不能被植物吸收利用。然而溶磷微生物能够分泌有机酸来溶解土壤中难溶性磷,提高土壤中磷的利用率,促进植物生长,提高作物的产量和品质。[目的]探究高效解磷荧光假单胞菌CLW17菌株的pqqE和GDH基因的生理学功能。[方法]利用生物在线软件对2个基因编码蛋白进行生物信息学分析。利用同源重组技术分别获得pqqE和GDH基因缺失突变株(CLW17ΔpqqE,CLW17ΔGDH),并使用接合转移的方式获得回补菌株(ΔpqqE/pqqE,ΔGDH/GDH)。分别采用NBRIP培养基、钼锑抗比色法及高压液相色谱法(HPLC)对野生型、突变株及互补株的溶磷及产有机酸能力进行检测。[结果]pqqE和GDH基因编码氨基酸数目分别为390和803,均无信号肽。pqqE无跨膜结构域,而GDH预测有5个跨膜结构域。pqqE和GDH基因是CLW17菌株的溶磷相关基因,2个基因的缺失均使该菌株的溶磷能力显著下降,而回补株可以恢复溶磷能力。CLW17野生株能分泌多种有机酸,其中葡萄糖酸(gluconic acid,GA)含量最多,其次是乙酸;但敲除株产有机酸的能力明显降低...