异氟醚和依托咪酯对重症肌无力患者胸腺切除术Bcl-2的影响

目的 探讨异氟醚和依托咪酯对重症肌无力患者胸腺切除术血清Bcl-2的影响。 方法 40例重症肌无力胸腺切除术患者,随机分为异氟醚组和依托咪酯组,每组20例。采用ELISA法检测患者术前、术后血清Bcl-2水平的变化。 结果 异氟醚组血清Bcl-2术后低于术前(P<0.01),依托咪酯组血清Bcl-2水平术后高于术前(P<0.01)。两组血清Bcl-2水平术前比较差异无统计学意义(P>0.05),术后血清Bcl-2水平依托咪酯组高于异氟醚组(P<0.01)。 结论 异氟醚用于重症肌无力患者麻醉可降低Bcl-2水平,而依托咪酯则升高Bcl-2水平,异氟醚可能比依托咪酯更适合用于重症肌无力患者胸腺切除术的麻醉。     

鼻敏片对寒证变应性鼻炎豚鼠辅助性T细胞17/调节性T细胞平衡的影响

目的:探讨鼻敏片对寒证变应性鼻炎(allergic rhinitis,AR)豚鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)表达水平的影响及其作用机制.方法:将40只雄性豚鼠随机分为空白组、模型组、鼻敏片组和氯雷他定片组,以复方凉药+卵清蛋白致敏法建立寒证AR模型后给予相应的药物干预,治疗15 d后,取血清和...     

CD4^＋CD25^＋T细胞亚群在系统性红斑狼疮患者中的检测和意义

目的:探讨CD4+CD25+T细胞亚群在系统性红斑狼疮(SLE)发病中的作用.方法:采用流式细胞仪检测35例SLE患者(其中活动组20例)及18例健康对照组的外周血CD4+CD25+T细胞亚群的百分比及其CD4+CD25highT细胞与CD4+CD25intT细胞的比率,同时评估疾病活动指数(SLEDAI),分析两者的相关性.结果:活动组和非活动组SLE患者外周血总CD4+GD25+T细胞、CD4+CD25highT细胞和CD4+CD25intT细胞占CD4+T细胞的百分比与对照组相比,差异均无统计学意义(P>0.05),且与SLEDAI无相关性.活动组SLE患者外周血CD4+CD25highT细胞与CD4+CD25intT细胞的比率与非活动组及健康对照组相比较明显降低,差异有统计学意义(P<0.05),且与SLEDAI呈明显负相关.结论:活动性SLE患者外周血CD4+CD25highT细胞与CD4+CD25intT细胞的比率降低可能在SLE的发病中起到重要的作用.     

Role of Cytochrome P450 3A4 and 1A2 Phenotyping in Patients with Advanced Non‐small‐Cell Lung Cancer Receiving Erlotinib Treatment

Erlotinib is metabolized by cytochrome p450 (CYP) 3A and CYP1A. This study assessed CYP3A4 (midazolam) and CYP1A2 (caffeine) phenotyping in plasma and dried blood spots (DBS) for predicting the pharmacokinetics and toxicity of erlotinib in 36 patients with advanced NSCLC. On day 1, erlotinib 150 mg OD was initiated, and the two oral probe drugs midazolam (2 mg) and caffeine (100 mg) were added on day 1. Plasma and DBS were collected for erlotinib, OSI‐420 and probe drugs for up to 6 hr on day 1 and 2‐weekly up to week 10. Probe drugs, erlotinib and OSI‐420 were analysed using LC‐MS‐MS, and PK data were processed using population modelling. A high correlation was found between plasma and DBS concentrations for erlotinib (R² = 0.960, p < 0.0001), OSI‐420 (R² = 0.971, p < 0.0001), midazolam (R² = 0.995, p < 0.0001) and caffeine (R² = 0.968, p < 0.0001). Apparent oral caffeine clearance was significantly correlated with erlotinib clearance (R² = 0.33, p = 0.048), while midazolam clearance was not (R² = ?0.09, p = 0.596). Erlotinib clearance was lower in patients experiencing grade 2 or 3 rash as compared to patients experiencing grade 0 or 1 rash (3.15 versus 3.93 L/hr, p = 0.086 for Student's t‐test). The results suggest that probe drug phenotyping is unlikely to substitute therapeutic drug monitoring of erlotinib in patients with advanced NSCLC, but erlotinib PK sampling from DBS may replace more invasive venous sampling and facilitate TDM in patients with cancer.     

自身免疫性疾病患者CYP3A4、CYP2C8和CYP3A5基因多态性与羟氯喹不良反应/血药浓度的相关性研究

目的:研究自身免疫性疾病(AID)患者细胞色素P_(450)(CYP)酶的亚型CYP3A4、CYP2C8和CYP3A5的基因多态性与羟氯喹不良反应、血药浓度的相关性,为羟氯喹的临床个体化用药提供参考。方法:病例来自安徽医科大学第一附属医院风湿免疫科2017年7月-2018年8月收治的因长期(>6个月)服用羟氯喹(日剂量为200～400 mg)出现不良反应的77例AID患者,收集患者信息、血样和不良反应发生情况,按不良反应发生部位分为肝功能正常组和肝功能异常组、肾功能正常组和肾功能异常组、眼部正常组和眼部异常组,采用高效液相色谱法测定羟氯喹全血药物浓度,MassARRAY系统检测患者CYP3A4、CYP2C8和CYP3A5的基因型,通过χ~2检验分析不同基因型患者羟氯喹不良反应的发生差异,通过两独立样本t检验和单因素方差分析不同基因型患者羟氯喹血药浓度的差异。结果:CYP3A5 rs4646453位点在肾功能正常组与肾功能异常组的分布差异具有统计学意义(P<0.05),肾功能异常组患者中CC基因型较AA+AC基因型的发生率高;CYP2C8 rs10882526位点在肝功能正常组与肝功能异常组的分布差异具有统计学意义(P<0.05),肝功能异常组患者中等位基因G较等位基因A的发生率高,AG基因型较AA基因型的发生率高。77例AID患者CYP3A4、CYP2C8和CYP3A5的基因多态性与血药浓度无显著相关性。在亚组分析时,58例系统性红斑狼疮(SLE)患者中CYP2C8 rs10882521的GT、GG和TT基因型的平均血药浓度分别为514.1、735.3、785.9 ng/mL,其中GG和TT基因型明显高于GT基因型(P<0.05)。结论:AID患者携带CYP3A5 rs4646453 CC基因型服用羟氯喹肾功能异常的发生率更高,携带CYP2C8 rs10882526位点的等位基因G和AG基因型服用羟氯喹肝功能异常的发生率更高。SLE患者服用相同剂量羟氯喹,携带CYP2C8 rs10882521 GT基因型较其他基因型患者血药浓度低。     

CD4+ T cell depletion changes the cytokine environment from a TH1/TH2 response to a TC17-like response in a murine model of atopic dermatitis

Atopic dermatitis (AD) is a common inflammatory skin disease often associated with co-morbidities including allergic hypersensitivity. We have studied induced AD-like disease in NC/Nga mice using the hapten FITC. Following FITC-treatment the NC/Nga mice develop AD-like skin lesions in regard to the histopathological and immunological changes. Consistent with AD in humans the number of CD4(+) T cells within the blood and draining lymph nodes increases considerably. To evaluate the contribution of T(H) cells on disease development we examined the effect of CD4 depletion. Following CD4 depletion the mice still develop AD-like skin lesions characterized by e.g. increased epidermal proliferation, hyperkeratosis and cellular infiltrate, however, the underlying immunological mechanisms change. CD4 depletion results in increased IL-17A and IL-22 production, which traditionally are associated with T(H)17 cells. Using confocal microscopy, we demonstrate that epidermal CD8(+) cells are positive for IL-17A, indicating that these cells are T(C)17 cells, the cytotoxic T cell counterpart to T(H)17 cells. In conclusion, we show that NC/Nga mice develop AD-like disease following CD4 depletion. This is mirrored by an increased production of IL-17A, which we suggest are produced by T(C)17 cells. These findings support that CD8(+) T cells can play a role in AD.     

MMP-9基因-1562 C>T多态性与2型糖尿病足溃疡的关联研究

目的：探讨基质金属蛋白9(matrix metalloproteinase-9, MMP-9)基因启动子单核苷酸多态性(-1562 C>T)与2型糖尿病(type 2 diabetes mellitus, T2DM)引起糖尿病足溃疡(diabetic foot ulcers, DFUs)的相关性。方法选择2010年7月—2012年12月就诊的无并发症的T2DM患者249例作为无并发症T2DM组,DFUs患者110例作为DFUs组,以及同期来医院进行健康体检的人员267例为对照组。应用聚合酶链反应( polymerase chain reaction, PCR)技术对MMP-9基因-1562C>T多态性突变区域进行扩增,并通过PCR-RFLP法检测MMP-9基因-1562C>T的CT、TT、CC基因型的分布,探讨与T2DM引起DFUs的相关性。所有的数据均采用IBM SPSS Statistics 20软件进行分析。结果3组MMP-9基因-1562 C>T基因位点发现C、T等位基因和CT、TT、CC及CT+TT 基因型,不同基因型出现频率在DFUs组、无并发症T2DM组以及对照组间的分布差异有统计学意义(χ2=21.69,P<0．05)。以CC型作为对照,CT/TT型在无并发症T2DM组与对照组间比较,差异有统计学意义(OR=1.82,P=0．002,95%CI：1.26,2.65);DFUs组与健康对照组相比,差异也有统计学意义(OR=1.98,P=0．005,95%CI：1.24,3.16)。无并发症T2DM组、DFUs组和健康对照组MMP-9基因-1562 C>T的T等位基因出现频率比较,差异有统计学意义( OR=1.87,P<0．001,95%CI：1.38,2.53;OR=2.00,P=0．001,95%CI：1.35,2.98)。结论 MMP-9的异常表达对DFUs的形成以及慢性蔓延有重要作用,是足部溃疡难以愈合的主要原因之一。     

Multidrug Resistance Associated Protein 1 together with Glutathione Plays a Protective Role against 4-Hydroxy-2-nonenal-Induced Oxidative Stress in Bovine Aortic Endothelial Cells

4-Hydroxy-2-nonenal (HNE), an aldehyde produced by lipid peroxidation, induces cytotoxicity and oxidative stress. Glutathione (GSH) protects against the cytotoxicity of HNE. However, the protective mechanism of GSH has not been fully examined. We examined the protective role played by the relationship between GSH and multidrug resistance associated protein 1 (MRP1) against the HNE-induced oxidative stress in bovine aortic endothelial cells (BAECs). HNE induced the loss of viability of BAECs. Exogenous GSH, which is membrane-impermeable, prevented the loss of viability induced by HNE by inhibiting HNE uptake in BAECs, probably due to the formation of the HNE-SG complex in the extracellular space. We demonstrated that HNE induced the expression of MRP1 protein, which can transport the HNE-SG complex. The induction of MRP1 protein expression by HNE disappeared in BAECs pretreated with L-buthionine sulfoximine, a GSH-depleting agent. This result suggests that HNE, together with intracellular GSH, contributes to the regulation of MRP1 protein expression. Moreover, we found that MK571, an MRP1 inhibitor, promoted the HNE-induced oxidative stress and cell death. Taken together, these findings suggest that MRP1, together with GSH, plays a protective role against the HNE-induced oxidative stress in BAECs.     

Metabolism of Dextromethorphan in vitro: Involvement of Cytochromes P450 2D6 AND 3A3/4, with a Possible Role of 2E1

Dextromethorphan (DMO), a cough suppressing synthetic analog of codeine, undergoes parallel O-demethylation to dextrorphan (DOP), and N-demethylation to 3-methoxy-morphinan (MEM), in humans. 3-hydroxymorphinan, a didemethylated metabolite, is formed secondarily. O-demethylation activity is well established as an index reaction for CYP2D6. However, this pathway appears to be mediated by at least two different enzymes in vitro. N-demethylation activity has recently been proposed to reflect CYP3A3/4 activity. We investigated both pathways in vitro with microsomal preparations from three human livers to assess the value of DMO as a probe drug for CYP2D6 and CYP3A3/4. DMO O-demethylation displayed a biphasic pattern with a high-affinity site reflecting CYP2D6 activity (mean K_i for quinidine, 0·1 ± 0·13 μM). Kinetic parameters for the two O-demethylation mediating enzymes predict an average relative intrinsic clearance (V_max/K_m ratio) of 96% of total O-demethylation mediated via the high-affinity enzyme. Thus, in vitro data confirms the usefulness of DMO O-demethylation as an index reaction to monitor CYP2D6 activity. The Eadie–Hofstee plot of DMO N-demethylation was consistent with single-enzyme Michaelis–Menten kinetics (V_max varying from 3·3 to 6·8 nmol mg⁻¹ min⁻¹, K_m from 231 to 322 μM). However, ketoconazole, a CYP3A3/4 inhibitor, reduced N-demethylation only by 60% and had a mean K_i an order of magnitude higher (0·37 μM) compared to other pure CYP3A3/4 mediated reactions. Troleandomycin, a mechanism based CYP3A3/4 inhibitor, inhibited MEM formation by an average maximum of 46%, with an IC₅₀ varying from 1 to 2·6 μM. A polyclonal rat liver CYP3A1 antibody inhibited MEM formation only by approximately 50%. Diethyldithiocarbamate (DDC), a mechanism based CYP2E1 inhibitor, reduced MEM formation at concentrations up to 150 μM between 33 and 43%. Chemical inhibitors of CYP2D6 (quinidine), CYP1A1/2 (α-naphthoflavone), and CYP2C9 (sulfaphenazole), as well as a goat rat liver CYP2C11 polyclonal antibody (inhibitory against human CYP2C9 and CYP2C19), had minimal effect on MEM formation rate, thus excluding an involvement of any of these enzymes. DMO N-demethylation is only partly mediated by CYP3A3/4, and therefore is not a reliable index reaction for CYP3A3/4 activity either in vitro or probably in vivo. © 1997 John Wiley & Sons, Ltd.