Meta-analysis of Cytochrome P4501A1 MspI Gene Polymorphism and Childhood Acute Leukemia

Objective To investigate the relationship between cytochrome P4501A1 （CYPIA1） Msp I gene polymorphism and childhood acute leukemia （AL）. Methods Relevant literature was extensively searched and screened by Pubmed and Wanfang Database, Chinese Science Journal Database and Chinese Journal Net. Various data consolidation, combined OR values and their 95% CI were tested by RevMan 4.2; Funnel plots were used for the bias analysis. Results Six related literatures were found to meet the requirements. According to heterogeneity results, there was no significant difference in homozygous types（P〉0.05）, while there was significant difference in two others types （P all〈0.05）. For wild CYPIAIMspl homozygous for the reference group, Combined OR of heterozygous mutation, homozygous, heterozygous ＋ homozygous mutation in AL and control groups were 1.18, 0.96, and 1.10 respectively. Subgroup analysis： Z values of CYP1A1Mspl homozygous, heterozygous ＋ homozygous in the acute lymphoblastic leukemia （ALL） and the control group were 0.10 and 0.76 respectively, Z values in non-acute lymphoblastic leukemia and control group were 0.74 and 0.75. Conclusion There is no correlation between CYP1A1Mspl gene polymorphism and the susceptibility of childhood AL.

The benzo(a) pyrene-induced mRNA expression of aromatic hydrocarbon receptor and cytochrome P4501A1 genes in rat liver

Objective To study the benzo(a)pyrene(B[a]P)-induced mRNA expression of aromatic hydrocarbon receptor(AHR)and cytochrome P4501A1(CYP1A1)genes in rat liver.Methods Rats were injected intraperitoneally with 5,10 and 15mg/kg of B[a]P.The total RNAs were extracted from rat livers by RNA purification kit,and the mRNA expression of AHR and CYP1A1 genes was determined by reverse transcription polymerase chain reaction(RT-PCR).β-actin was used as the internal control.The mRNA expression of both AHR and CYP1A1 genes was measured at indicated time points(24,48 and 72h)after B[a]P treatment at three different concentrations(5,10 and 15mg/kg).Results The mRNA expression of AHR gene increased in a time-dependent manner at the concentration of 10mg/kg but not at 5 and 15mg/kg of B[a]P.The mRNA expression of CYP1A1 gene differed significantly at 48h and 24h in rat livers treated with 10 and 15mg/kg dosage of B[a]P.The mRNA expression of AHR and CYP1A1 genes increased with B[a]P treatment in a concentration-dependent manner.The time-dependent increase in mRNA expression was shown by AHR but not by CYP1A1 gene with B[a]P(10mg/kg)treatment.Conclusion This study demonstrates that toxic B[a]P increases the mRNA expression of both AHR and CYP1A1 genes in vivo,suggesting that B[a]P may play a role in cancer genesis by this way.

饮酒和CYP1A1-MspI、ALDH-2基因多态性与食管癌的相关性

目的探讨饮酒和细胞色素P4501A1-MspI（CYP1A1-MspI）、乙醛脱氢酶-2（ALDH-2）基因多态性与食管癌发病之间的关系。方法采用病例-对照研究的方法,以160例食管癌患者及160例非癌对照者的外周血白细胞为样本,利用聚合酶链反应（PCR）技术分析了CYP1A1-MspI和ALDH-2基因多态性。结果 CYP1A1-MspI突变纯合型（m2/m2）和ALDH-2变异基因型频率分布分别为39.375%、70.625%（病例组）和20.000%、43.750%（对照组）,二者差异显著（P〈0.01,P〈0.01）。CYP1A1-MspI（m2/m2）患食管癌的风险显著增加（OR=2.598,95%CI=1.819~4.265）。ALDH-2变异基因型者患食管癌的风险也显著增加（OR=3.091,95%CI=1.922~4.738）。基因突变的协同分析发现CYP1A1-Ms-pI（m2/m2）/ALDH-2变异基因型者在食管癌组和对照组中的分布频率分别为31.875%和6.250%,二者有显著差异（P〈0.01）。CYP1A1-MspI（m2/m2）/ALDH-2变异基因型者患食管癌的风险显著增加（OR=9.909,95%CI=3.574~12.532）。病例组的饮酒率显著高于对照组的饮酒率（OR=3.096,95%CI=1.532~4.88 0,P〈0.01）,CYP1A1-MspI（m2/m2）/及ALDH-2变异基因型与饮酒有协同作用（OR=40.727,95%CI=17.965~66.572）。结论 CYP1A1-MspI（m2/m2）/ALDH-2变异基因型和饮酒是食管癌的易患因素,三者的联合在食管癌的发生中起着协同的作用。

辛硫磷对鲫鱼肝微粒体中CYP1A活性及其表达水平的影响

【目的】研究辛硫磷暴露对鲫鱼肝微粒体中细胞色素P4501a(CYP1A)活性及其mRNA和蛋白表达的影响,明确其剂量—效应及时间—效应关系,为揭示有机磷农药对水生生物的代谢调节机制提供理论依据。【方法】设辛硫磷低、中、高浓度组(0.0825、0.165和0.33 mg/L)和丙酮(助溶剂)对照组,采用半静态染毒法染毒鲫鱼,分别于染毒24、48、72和96 h后取样并迅速剖解取出鲫鱼肝胰脏,以CYP1A相关酶7-乙氧基-3-异吩唑酮-脱乙基酶(EROD)活性反映CYP1A活性,采用实时荧光定量PCR和Western blotting分别测定鲫鱼肝微粒体中CYP1A基因mRNA及其蛋白的表达情况。【结果】辛硫磷对鲫鱼肝微粒体中CYP1A活性产生抑制作用,辛硫磷染毒24 h时CYP1A活性与辛硫磷存在明显的剂量—效应关系;各辛硫磷浓度组间均存在明显的时间—效应关系,至染毒96 h时低、中、高浓度组鲫鱼肝微粒体中CYP1A活性分别较对照组下降54.7%、30.4%和65.5%,且差异极显著(P<0.01,下同)。辛硫磷染毒后,鲫鱼肝微粒体中CYP1A基因mRNA的表达整体上呈明显下调趋势,各染毒时间组间均存在较强的剂量—效应关系,除染毒48 h时CYP1A基因mRNA相对表达量与辛硫磷浓度呈正相关外,其他染毒时间点均呈负相关。辛硫磷对鲫鱼肝微粒体中CYP1A蛋白表达的影响均达极显著水平,且呈明显的剂量—效应及时间—效应关系,至染毒96 h时低、中、高浓度组的表达量分别较对照组降低74.6%、82.6%和85.7%。【结论】辛硫磷能抑制鲫鱼肝微粒体中CYP1A活性并下调CYP1A基因mRNA及其蛋白的表达;相对于辛硫磷产生的剂量—效应影响,其对CYP1A活性及其蛋白表达的时间—效应影响更明显。

饮水型砷暴露地区人群血液CYP1A1mRNA表达对砷中毒患病情况的影响

目的:通过分析饮水型砷暴露人群血液中CYP1A1mRNA的表达情况,探讨CYP1A1mRNA的表达与高砷暴露地区砷中毒患病情况之间的关系。方法:选取内蒙古自治区巴彦淖尔市的砷暴露地区作为调查点,将在此居住10年以上的居民作为调查对象。按饮水砷浓度将233名居民分为4组:对照组(饮水砷浓度<10.0μg/L),低剂量组(饮水砷浓度10.0~99.9μg/L),中剂量组(饮水砷浓度100~199.9μg/L),高剂量组(饮水砷浓度≥200.0μg/L)。采用流行病学调查结合实时荧光定量PCR检测技术,测定血液CYP1A1 mRNA的表达水平,分析基因表达与砷中毒患病情况之间的关系。结果:(1)砷中毒的患病率随水砷浓度升高逐渐升高,与对照组相比,各剂量组人群砷中毒患病存在上升趋势,差异有统计学意义(χ~2=16.97,P<0.05),经线性趋势卡方检验发现随着水砷浓度的升高,砷中毒的病情逐渐加重(χ~2=2.371,P<0.05);(2)长期砷暴露会导致CYP1A1mRNA的表达水平升高,高浓度组CYP1A1mRNA表达水平升高具有统计学意义(P<0.05);(3)随着砷中毒的病情加重,各组之间的CYP1A1mRNA的表达水平没有显著性差异(P>0.05)。结论:长期慢性砷暴露会导致CYP1A1mRNA的表达水平升高,这可能是影响地方性砷中毒患病的重要因素,但CYP1A1mRNA的表达与砷中毒病情严重程度无关。

细胞色素P450 1A1的研究进展

细胞色素P450酶系参与生物体内许多内源性及外源性物质的生物转化。其中细胞色素 P4501A1(CYP1A1)广泛存在于多种肝外组织,参与了多种前致癌物和致突变物的活化代谢过程,并与 多种肿瘤发生相关。本文综述了CYP1A1在体内的分布、诱导和抑制,遗传多态性及癌症易感性,简要 介绍了该领域研究的进展情况。

鸡细胞色素P450 1A5的原核表达与活性重组

旨在对鸡细胞色素P450 1A5(CYP1A5)蛋白进行体外功能研究,采用大肠杆菌系统进行CYP1A5的异源表达。以鸡的cDNA为模板,扩增出CYP1A5基因,将该基因的N端编码区进行修饰,并连接到pCW载体中构建His-CYP1A5,经IPTG诱导在大肠杆菌中表达。经CO-差示光谱检测,所获得的His-CYP1A5具有典型的P450吸收峰。该蛋白与细胞色素P450还原酶(CPR)进行体外重组,构成的重组酶系表现出乙氧基试卤灵-O-脱乙基酶活性。结果表明,所采用的表达策略可以成功产生出具有催化活性的鸡细胞色素P450 1A5(CYP1A5)蛋白。

Sensing cytochrome P4501A1 activity by a resorufin-based isoform-specific fluorescent probe

Cytochrome P4501 A1(CYP1A1),a heme-containing monooxygenase,is of particular importance for human health because of its vital roles in the metabolic activation of pro-carcinogenic compounds to the carcinogens.Deciphering the relevance of CYP1A1 to human diseases and screening of CYP1A1 modulators require reliable tool(s)for probing this key enzyme in complex biological matrices.Herein,a practical and ultrasensitive fluorescence-based assay for real-time sensing CYP1A1 activities in biological systems has been developed,via designing an isoform-specific fluorogenic sensor for CYP1A1(CHPO).The newly developed fluorogenic substrate for CYP1A1 has been carefully investigated in terms of specificity,sensitivity,precision,quantitative linear range and the anti-interference ability.The excellent selectivity,strong anti-interference ability and fast response kinetics,making the practicability of CHPObased CYP1A1 activity assay is better than that of most reported CYP1A1 activity assays.Furthermore,CHPO has been successfully used for imaging CYP1A1 activities in living cells and human tissues,as well as for high-throughput screening of CYP1A1 inhibitors using tissue preparations as enzyme sources.Collectively,this study provided a practical fluorogenic sensor for real-time sensing CYP1A1 in complex biological systems,which would strongly facilitate the investigations on the relevance of CYP1A1 to human diseases and promote high-throughput screening of CYP1A1 modulators for biomedical applications.

Functional reconstruction of bovine P450scc steroidogenic system in <i>Escherichia coli</i>

Mammalian cytochrome P450scc enzyme system catalyzes the initial step in steroid hormone biosynthesis—cholesterol hydroxylation followed by cleavage of the side-chain to yield pregnenolone. This system consists of three components—the cytochrome P450scc (CYP11A1), a flavoprotein (NADPH-adrenodoxin reductase, AdR) and an iron-sulfur protein (adrenodoxin, Adx). In this work, the three-component electron transport chain (AdR/Adx/CYP11A1) from bovine adrenal cortex has been implemented in Escherichia coli by co-expression of the corresponding coding sequences from a tricistronic plasmid. The cDNAs of AdR, Adx and CYP11A1 are situated in a single transcription unit and separated by ribosome binding sequences. The recombinant strain created was capable of synthesizing functional proteins identical to the bovine CYP11A1, AdR and Adx on molecular weights and immuno-specificity. The experiments in vivo showed pregnenolone production from cholesterol by the transformed bacteria. Maximal productivity of 0.42 ± 0.015 mg/l pregnenolone for 24 h has been reached for the induced cells in the presence of cholesterol solubilizing agent—methyl-β-cyclodextrin. Thus, a stable transgenic E. coli strain with the functional reconstructed bovine cholesterol side-chain cleavage system has been firstly generated in this work. The findings are of importance for studies of mammalian steroidogenic system features, and may open some perspectives for further generation of novel microbial biocatalysts.

CYP1A1及GSTP1基因多态性与肺癌易感性关系

目的探讨细胞色素P4501A1(CYP1A1)Exon7和谷胱甘肽硫转移酶P1(GSTP1)Ile105Val基因多态性与内蒙古地区汉族人群肺癌易感性关系。方法采用等位基因特异性扩增法分析216例汉族对照人群和116例肺癌患者CYP1A1 Exon7和GSTP1 Ile105Val基因多态性。结果携带CYP1A1 Exon7突变杂合型和纯合型的个体患肺癌的危险均升高(OR值分别为1.460和1.593),而携带GSTP1 Ile105Val突变杂合型和纯合型的个体患肺癌的风险均降低(OR值分别为0.970和0.602);CYP1A1 Exon7和GSTP1 Ile105Val基因在肺癌易感性方面无协同作用;CYP1A1 Exon7与吸烟有协同作用(OR=2.637,95%CI=1.056～6.530,P=0.032),GSTP1 Ile105Val与吸烟无协同作用。结论 CYP1A1 Exon7突变基因型为肺癌的可疑易感因素,CYP1A1 Exon7突变基因型和吸烟对肺癌易感有协同作用,GSTP1 Ile105Val突变基因型可降低肺癌易感性。

Identification of a novel cytochrome P450 CYP321B1 gene from tobacco cutworm （Spodoptera litura） and RNA interference to evaluate its role in commonly used insecticides

Insect cytochrome P450 monooxygenases （CYPs or P450s） play an important role in detoxifying insecticides leading to resistance in insect populations. A polyphagous pest, Spodoptera litura, has developed resistance to a wide range of insecticides. In the present study, a novel P450 gene, CYP321B1, was cloned from S. litura. The function of CYP321B1 was assessed using RNA interference （RNAi） and monitoring resistance levels for three commonly used insecticides, including chlorpyrifos, β-cypermethrin and methomyl. The full-length complementary DNA sequence of CYP321B1 is 1814 bp long with an open reading frame of 1 488 bp encoding 495 amino acid residues. Quantitative reverse-transcriptase polymerase chain reaction analyses during larval and pupal develop- ment indicated that CYP321B1 expression was highest in the midgut of fifth-instar larvae, followed by fat body and cuticle. The expression of CYP321B1 in the midgut was up- regulated by chlorpyrifos,β-cypermethrin and methomyl with both lethal concentration at 15% （LC15） （50, 100 and 150 μg/mL, respectively） and 50%（LC50） dosages （100,200 and 300μg/mL, respectively）. Addition of piperonyl butoxide （PBO） significantly increased the toxicity ofchlorpyrifos,β-cypermethrin and methomyl to S. litura, suggesting a marked synergism of the three insecticides with PBO and P450-mediated detoxification. RNAi- mediated silencing of CYP321B1 further increased mortality by 25.6% and 38.9% when the fifth-instar larvae were exposed to chlorpyrifos and β-cypermethrin, respectively, at the LCso dose levels. The results demonstrate that CYP321B1 might play an important role in chlorpyrifos and β-cypermethrin detoxification in S. litura.

Preferential expression of cytochrome CYP CYP2R1 but not CYP1B1 in human cord blood hematopoietic stem and progenitor cells

Cytochrome P450(CYP)enzymes metabolize numerous endogenous substrates,such as retinoids,androgens,estrogens and vitamin D,that can modulate important cellular processes,including proliferation,differentiation and apoptosis.The aim of this study is to characterize the expression of CYP genes in CD34+human cord blood hematopoietic stem and early progenitor cells(CBHSPCs)as a first step toward assessment of the potential biological functions of CYP enzymes in regulating the expansion or differentiation of these cells.CD34+CBHSPCs were purified from umbilical cord blood via antibody affinity chromatography.Purity of CD34+CBHSPCs was assessed using fluorescence-activated cell sorting.RNA was isolated from purified CD34+CBHSPCs and total mononuclear cells(MNCs)for RNA-PCR analysis of CYP expression.Fourteen human CYPs were detected in the initial screening with qualitative RT-PCR in CD34+CBHSPCs.Further quantitative RNA-PCR analysis of the detected CYP transcripts yielded evidence for preferential expression of CYP2R1 in CD34+CBHSPCs relative to MNCs;and for greater expression of CYP1B1 in MNCs relative to CD34+CBHSPCs.These findings provide the basis for further studies on possible functions of CYP2R1 and CYP1B1 in CBHSPCs'proliferation and/or differentiation and their potential utility as targets for drugs designed to modulate CD34+CBHSPC expansion or differentiation.

A novel cytochrome P450 gene from Catharanthus roseus cell line C_(20)hi:cloning and characterization of expression

An expressed sequence tag(EST)obtained from a subtractive-suppression hybridization cDNA library constructed using Catharanthus roseus cell line C_(20)hi and its parental cell line C_(20)D was used to clone a ful-length cytochrome P450 cDNA of cyp71d1.The encoded polypeptide contained 507 amino acids with 39-56% identity to other CYP7ID subfamily members at the.amino acid level.Expression characteristics of cyp71d1 were determined using semi-quantitative RT-PCR.The cyp71d1 transcript was expressed in all three cell lines with the highest level in the cell line C_(20)hi.In the mature C.roseus plant,the cyp71d1 cDNA was highly expressed in petals,roots and stems,but very weakly expressed in young leaves.Its transcription level increased with the development of flowers.2,4-D could down-regulate the transcription of cyp71d1,as did KT,but only to a minor degree.Neither light nor yeast elicitor could induce the transcription of cyp71d1.

细胞色素P4501 A1和谷胱苷肽硫转移酶基因多态性与肺癌关系的病例对照研究

目的 探讨细胞色素P4 5 0 1A1(CYP 1A1)和谷胱苷肽硫转移酶 (GST) M1基因多态性与肺癌易感性的关系。方法 选取新发肺癌患者 91例及同期非肺部疾患同性别患者 91例作匹配 ,另选取体检正常者 4 7例做频数对照 ,采用聚合酶链式反应 (PCR)和限制性片段长度多态性 (RFLP)技术检测CYP 1A1和GST M1的基因多态性。结果 单独分析CYP 1A1和GST M1基因多态性与肺癌的关系 ,其OR值分别为 1.5 3和 1.4 2 ,与对照组比较 ,差异均无显著性 (P >0 .0 5 ) ,表明与肺癌的发生无相关性。而将二者联合分析时 ,其OR值为 2 .4 7,95 %CI为 1.0 3～ 5 .90 ,与对照组比较 ,差异有显著性(P <0 .0 5 ) ,表明与肺癌的发生有一定相关性。结论 CYP 1A1和GST M1的单一基因多态性不增加患肺癌的危险 ,而两者联合作用时 ,则可增强患肺癌的风险。

CYP1B1对肥胖小鼠脂肪组织血管新生因子影响

目的探讨血管新生因子血管生成素Ⅰ和Ⅱ在保护幼年细胞色素P4501B1(CYP1B1)基因敲除小鼠营养性肥胖中作用。方法 CYP1B1基因敲除和野生型雄性C57/BL小鼠(3周龄)各16只,给予低脂膳食(10%脂肪)、高脂膳食(60%脂肪)饲料11周;小鼠处死后取附睾脂肪组织检测血管密度及血管新生因子基因和蛋白表达。结果野生高脂组小鼠脂肪组织血管密度下降,基因敲除小鼠脂肪组织血管分布不受影响;高脂膳食诱导下,野生型和基因敲除小鼠血小板-内皮细胞粘附分子CD31 mRNA表达下调(P<0.05),野生型小鼠CD31蛋白表达下调(P<0.05);与野生低脂组比较,高脂诱导后野生型和基因敲除小鼠血管生成素Ⅰ表达量分别为0.35和0.50(P<0.05),瘦素表达量则分别为2.48和1.42(P<0.05);敲除高脂组瘦素表达量较野生高脂组下降(P<0.05)。结论CYP1B1基因敲除对血管新生相关因子的调控可能在其营养性肥胖中起一定保护作用。

CYP2E1基因多态性与客家人群胃癌易感性的研究

目的：通过研究细胞色素P4502E1（CYP2E1）基因多态性位点rs2031920与客家人群胃癌遗传易感性的相关性，探讨遗传和环境因素在胃癌发病中的作用。方法采取病例-对照研究，选择经胃镜和病理检查确诊的梅州地区客家人群51例胃癌患者（胃癌组）和52例正常对照（对照组），对CYP2E1 rs2031920（C-1053T）位点进行基因型及等位基因检测，分析其在两组间的分布特征。结果CYP2E1 rs2031920位点在梅州地区客家人群中存在 CC、CT、TT 多态性，各基因型在胃癌组的分布频率为62．75％（32／51）、33．33％（17／51）、3．92％（2／51），在对照组的分布频率为59．62％（31／52）、34．61％（18／52）、5．77％（3／52），两组之间的差异无统计学意义（χ2＝0．235，P ＝0．889）。CYP2E1基因rs2031920位点的等位基因 C、T 在胃癌组和对照组构成比分别为79．41％（81／102）、20．59％（21／102）和76．92％（80／104）、23．08％（24／104），差异无统计学意义（χ2＝0．186，P ＝0．666）。经性别、年龄分层分析结果显示也无统计学意义（χ2＝4．412，P ＝0．129；χ2＝0．898，P ＝0．473）。结论 CYP2E1的基因多态性位点 rs2031920与客家人群胃癌的易感性不相关。

苗族人群细胞色素及GSTM1基因多态性分布

目的了解细胞色素P4501A1(CYP1A1)与谷胱甘肽硫转移酶(GST)M1基因多态性在贵州苗族人群中的分布。方法采用聚合酶链反应(PCR)和聚合酶链反应-限制性片断长度多态性(PCR-RFLP)技术检测CYP1A1和GSTM1基因的多态性。结果97人中CYP1A1基因型为野生型、杂合型、突变型的分别有69,18,10人,分别占71.13%,18.56%,10.31%;野生型等位基因频率为0.805,突变型等位基因频率为0.195。97人中有67人为GSTM1基因缺陷型,GSTM1缺陷型占69%。结论贵州苗族CYP1A1m1突变型等位基因频率明显低于亚洲人,而高于欧洲人群;贵州苗族GSTM1基因缺陷率与中国汉族人相比无明显差异,但明显高于日本人、韩国人、德国人和波兰人。

葛根素对人细胞色素P4502D6酶活性的影响及其与基因型关系的研究

目的：研究葛根素对人细胞色素P450（ CYP）1D6酶活性的影响及其与CYP1D6基因型的关系。方法（1）体外试验：取肝内胆管结石患者手术切除标本中正常肝组织制备人肝微粒体孵育体系，加入不同浓度葛根素（0、0.015、0.050、0.100、0.100、0.400、0.800 mmol/L）及美托洛尔，以高效液相色谱法检测美托洛尔代谢产物α-羟基美托洛尔产率，以此反映CYP1D6酶活性。（1）体内试验：以18名健康男性[年龄19~16（11±4）岁，体重61~75（69±7）kg]为对象，其中CYP1D6基因型为＊1/＊1、＊1/＊10、＊10/＊10者各6名。试验分3个阶段进行，采用两阶段交叉试验设计。将受试者按照简单随机化分组方法随机分为1组。第1阶段（第1~11天）：1组连续10 d静脉滴注葛根素注射液400 mg/d，另1组不给药，第11天清晨1组受试者均口服美托洛尔100 mg；第1阶段（第11~17天）：1组均不采取任何措施；第3阶段（第18~18天）：第1阶段未服药组连续10 d静脉滴注葛根素注射液400 mg/d，另1组不给药，第18天清晨1组受试者均口服美托洛尔100 mg。第11和18天口服美托洛尔100 mg后收集受试者连续8 h尿液，采用高效液相色谱法测定尿液中美托洛尔及α-羟基美托洛尔浓度，以二者之比反映CYP1D6酶活性。结果体外试验显示经0、0.015、0.050、0.100、0.100、0.400、0.800 mmol/L葛根素处理的人肝微粒体反应体系α-羟基美托洛尔产率分别为（0.018±0.001）、（0.015±0.001）、（0.015±0.001）、（0.011±0.001）、（0.011±0.001）、（0.010±0.001）、（0.005±0.001）mmol/（mg·h），α-羟基美托洛尔产率随着葛根素浓度的升高而逐渐降低，总体均数间差异有统计学意义（ F =30.750，P =0.000）。两两比较显示，0.100、0.100、0.400和0.800 mmol/L葛根素组α-羟基美托洛尔产率明显低于0、0.015、0.050 mmol/L葛根素组（ P﹤0.05），0.100、0.400、0.800 mmol/L 葛根素组明显低于0.015、0.050 mmol/L 葛根素组（ P ﹤0.05），而0.800 mmol/L葛根素组明显低于0.100、0.100、0.400 mmol/L葛根素组（ P﹤0.05）。体内试验显示18名受试者应用葛根素前后尿样中美托洛尔与α-羟基美托洛尔浓度比值分别为1.11±0.55和1.73±0.94，差异有统计学意义（ P﹤0.01）。不同CYP1D6基因型的3组受试者应用葛根素前后尿样中CYP 1D6活性变化程度组间比较，差异无统计学意义。结论葛根素对CYP1D6活性有明显的抑制作用，浓度越高抑制作用越明显，但与CYP1D6基因型无明显相关性。

Single-cell RNA-seq unveils critical regulators of human FOXP3^+regulatory T cell stability

The heterogeneity and plasticity of T lymphocytes is critical for determining immune response outcomes.Functional regulatory T(Treg)cells are commonly characterized by stable FOXP3 expression and have reported to exhibit heterogeneous phenotypes under inflammatory conditions.However,the interplay between inflammation and Treg cell suppressive activity still remains elusive.Here,we utilized singlecell RNA sequencing to investigate how human Treg cells respond to the pro-inflammatory cytokine interleukin-6(IL-6).We observed that Treg cells divided into two subpopulations after IL-6 stimulation.TIGITàunstable Treg cells lost FOXP3 expression and gained an effector-like T cell phenotype,whereas TIGIT+Treg cells retained robust suppressive function.Single cell transcriptome analysis revealed a spectrum of cellular states of IL-6-stimulated Treg cells and how cytochrome P450 family 1 subfamily A member 1(CYP1A1)is a crucial regulator of Treg cell suppressive capability and stability.CYP1A1-deficient human Treg cells developed a Th17-like phenotype after IL-6 stimulation.Our findings implicate CYP1A1 as a previously unidentified regulator of Treg cells that may have target potential for clinical application for biotherapies.

Hepatoprotective and antioxidant effects of dietary Glycyrrhiza polysaccharide against TCDD-induced hepatic injury and RT-PCR quantification of AHR2,ARNT2,CYPIA mRNA in Jian Carp(Cyprinus carpio var.Jian)

To evaluate the protective effects of Glycyrrhiza polysaccharide（GPS） against 2,3,7,8-tetrachlorodibenzo-p-dioxin（TCDD）-induced hepatotoxicity in Jian carp,the fish were fed diets containing GPS at doses of 0.1,0.5 and 1.0 g/kg for 60 days before an intraperitoneal injection of 0.6 μg/kg TCDD at a volume of 0.05 mL/10 g body weight.At 72 hr post-injection,blood and liver samples were taken for biochemical analysis and the fish liver samples were used for the preparation of pathological slices.The results showed that increases in alanine aminotransferase（GPT）,aspartate aminotransferase（GOT）,lactate dehydrogenase（LDH）,and alkaline phosphatase（AKP） in serum induced by TCDD were significantly inhibited by pre-treatment with 1.0 g/kg GPS.Following the 1.0 g/kg GPS pre-treatment,total protein（TP）,albumin（Alb）,catalase（CAT）,glutathione peroxidase（GPx）,total antioxidant capacity（T-AOC） and superoxide dismutase（SOD） activities in liver tissue increased significantly,malondialdehyde（MDA） formation（P ＆lt; 0.05 or P ＆lt; 0.01） was significantly inhibited,and the expression of cytochrome P4501A（CYP1A）,aryl hydrocarbon receptor 2（AHR2） and aryl hydrocarbon receptor nuclear translocator 2（ARNT2） mRNA（P ＆lt; 0.05） was significantly enhanced.Histological observations on fish liver were obtained by preparing paraffin tissue sections via HE staining,and the results showed that histological changes were obviously reduced by 0.5 and 1.0 g/kg GPS.GPS significantly reduced liver tissue damage caused by TCDD.Overall,these results proved the hepatoprotective effect of GPS in protecting against fish liver injury induced by TCDD,and supported the use of GPS（1.0 g/kg） as a hepatoprotective and antioxidant agent in fish.