肺纤维化鼠Ⅰ型前胶原和Ⅲ型前胶原mRNA表达及芪丹颗粒剂的干预效应

目的:观察芪丹颗粒对博莱霉素致肺纤维化鼠的Ⅰ型前胶原和Ⅲ型前胶原mRNA表达的影响,探讨其作用机制。方法:实验于2004-10/2006-10在山东省医学科学院基础所病理实验室完成。实验材料:清洁级雄性SD大鼠160只,体质量180～210g。氢化可的松琥珀酸钠50mg/支;芪丹(颗粒剂)是由黄芪、丹参、川芎等中草药加工提纯后的粗提取物制成的颗粒剂,10g/包(1g干粉相当于原生药8g)。实验分组:160只SD大鼠按随机区组设计分为正常组20只、模型组40只、芪丹颗粒剂50只、氢化可的松50只。实验干预:正常组20只气管内灌注和灌胃均用生理盐水。其余大鼠经气管内一次性灌注博莱霉素A5按0.25mL左右(5mg/kg体质量)诱导大鼠肺间质纤维化。随机取40只为模型组。取芪丹颗粒剂组和氢化可地松组大鼠各30只,分别从造模后第2天灌注芪丹颗粒剂(3125mg/kg)和腹腔注射氢化可的松(25mg/kg),药物干预后第7,14,28天麻醉下处死动物。两组各余20只分别从造模14d后灌注芪丹颗粒剂和腹腔注射氢化可的松(用量同前),于第28、42天分别处死动物。实验评估:用苏木精-伊红评价肺组织病理学变化和原位杂交方法检测各组大鼠肺Ⅰ型和Ⅲ型前胶原mRNA表达。结果:160只大鼠全部进入结果分析。①大鼠肺脏大体标本观察:对照组肺组织各观察时间点无明显改变,模型组肺组织表面凸凹不平,部分肺叶体积缩小,表面见灰白色结节。氢化可的松组与模型组相似。芪丹颗粒剂组见部分肺叶表面不光滑及大小不等结节。②肺组织病理学观察:模型组7d肺泡腔内大量巨噬细胞淋巴细胞中性粒细胞浸润,肺间质成纤维细胞增殖,28d肺泡结构破坏肺泡内见大量胶原纤维和成纤维细胞。芪丹颗粒剂组肺泡炎及肺纤维化程度均明显轻于模型组和氢化可的松组(P<0.05)。③肺间质纤维化形成中Ⅰ型、Ⅲ型前胶原mRNA表达:原位杂交显示两种前胶原mRNA表达呈动态变化,早期肺泡炎以Ⅲ型前胶原mRNA大量增生为主,晚期纤维化期以Ⅰ型前胶原mRNA增生为主。芪丹颗粒剂组Ⅲ型前胶原mRNA的表达在第14天处于最高,至28d仍维持较高的水平,芪丹颗粒剂组Ⅲ型前胶原mRNA的表达高于模型组和氢化可的松组(P<0.05)。28d,Ⅰ型前胶原mRNA的表达在第二天给药芪丹颗粒剂组和氢化可地松组及第14天给药芪丹颗粒剂组和氢化可的松组组间差异均有显著性(P<0.05)。结论:大鼠肺纤维化的早期以Ⅲ型前胶原mRNA的表达为主,晚期纤维化期以Ⅰ型前胶原mRNA的大量表达为主。芪丹颗粒可减轻博莱霉素诱导的大鼠肺泡炎及肺纤维化的程度,其机制可能通过影响了Ⅰ型和Ⅲ型前胶原mRNA的代谢和表达,从而减慢肺间质纤维化的进程。芪丹颗粒对肺间质纤维化有治疗作用,且优于氢化可的松。     

大剂量免疫球蛋白预防肾移植术后肺部感染:随机对照

目的:肾移植术后半年内发生的并发症主要为感染,尤以肺部感染为重。静脉注射大剂量免疫球蛋白预防肾移植术后早期肺部感染的效果值得临床应用中予以总结提高。方法:①实验对象及分组:选择2002-01/2006-12于海南省人民医院肾内科就诊的肾移植术后半年内患者40例,对实验及治疗方案均知情同意,且得到医院伦理道德委员会批准。按随机数字表法分为2组,免疫球蛋白组、对照组各20例。②实验分组:免疫球蛋白组在应用免疫抑制剂基础上用0.4g/(kg·d)免疫球蛋白治疗;对照组应用免疫抑制剂。③实验评估:观察两组患者普通肺炎、重症肺炎发生率及血清IgG和T淋巴细胞亚群水平变化。结果:40例患者全部进入结果分析。①免疫球蛋白组患者的普通肺炎感染率及重症肺炎感染率均低于对照组(P<0.01)。②免疫球蛋白组患者血IgG、T淋巴细胞亚群CD3 ,CD4 ,CD8 水平均高于对照组(P<0.05),用免疫球蛋白治疗未见副作用。结论:随机对照结果显示,肾移植术后早期应用大剂量免疫球蛋白能明显减少肺部感染发生,减少并发症,提高肾移植患者生存率。     

Relationship between the development of blindness in Churg-Strauss syndrome and anti-myeloperoxidase antibodies

Occular involvement is infrequent and blindness rare in Churg-Strauss syndrome. We describe a patient with Churg-Strauss syndrome who presented with blindness. This was associated with the appearance of circulating autoantibodies to myeloperoxidase.      

Measurement of the quality of life in rheumatic disorders using the EuroQol

The EuroQol is a validated quality of life (QOL) scale that has been used in population and clinical studies, and has been reported in patients with rheumatoid arthritis (RA). It is short, simple to complete, and might be suitable for surveys of rheumatic disease patients. The properties of this instrument were investigated in a postal survey of 1372 rheumatic disease patients, including 537 with RA, 319 with osteoarthritis (OA) and 516 with fibromyalgia. In addition, simultaneous measurements of functional disability, pain, psychological status, global severity and demographic characteristics were made. EuroQol scores (0.57) were significantly lower than VAS health state scores (0.67) and arthritis-related global severity scores (0.62). QOL was similar in RA and OA, but lower in fibromyalgia, across all instruments. The distribution of EuroQol scores had many gaps and was not continuous. EuroQol did not reflect VAS QOL scores at EuroQol levels below 0.5, and the mean score difference between the instruments below that level was 0.43. Many patients with low EuroQol scores (including some with health states that were 'worse than death') had high VAS scores. These differences appear to have arisen because disability, pain and depression questions ask about mild or moderate problems, but not both, thereby forcing scale compression in the mid ranges. In addition, the 'severe' value is so extremely abnormal that few patients endorse it. Finally, penalty scores are applied to those with at least one maximally abnormal score. The scoring properties and distributional aspects of the EuroQol indicate substantial problems in its use in rheumatic disease patients.      

荷兰异波帕明多中心研究(DIMT)

标题　ａ）Ｉｂｏｐａｍｉｎｅ和地高辛治疗轻到中度的心力衰竭ｂ）在６个月的随访中轻度而未治的心衰之发展及ｉｂｏｐａｍｉｎｅ和地高辛单疗之效果作者　ａ）ＶａｎＶｅｌｄｈｕｉｓｅｎＤＪ,ｅｔａｌ－ＪＡｍＣｏｌｌＣａｒｄｉｏｌ,１９９３,２２：１５６４（英文）ｂ）ＶａｎＶｅｌｄｈｕｉｓｅｎＤＪ,ｅｔａｌ－ＡｍＪＣａｒｄｉｏｌ,１９９５,７５：７９６（英文）　　研究疾病：轻到中度的心力衰竭。目的：评价ｉｂｏｐａｍｉｎｅ的疗效和安全性,并将其与地高辛和安慰剂相比较。　　设计：随机、双盲和安慰剂对照。病人资料…     

Targeted inactivation of the major positive regulatory element (HS-40) of the human alpha-globin gene locus

We have examined the role of the major positive upstream regulatory element of the human alpha-globin gene locus (HS-40) in its natural chromosomal context. Using homologous recombination, HS-40 was replaced by a neo marker gene in a mouse erythroleukemia hybrid cell line containing a single copy of human chromosome 16. In clones from which HS-40 had been deleted, human alpha-globin gene expression was severely reduced, although basal levels of alpha 1 and alpha 2-globin mRNA expression representing less than 3% of the level in control cell lines were detected. Deletion of the neo marker gene, by using FLP recombinase/FLP recombinase target system, proved that the phenotype observed was not caused by the regulatory elements of this marker gene. In the targeted clones, deletion of HS-40 apparently does not affect long-range or local chromatin structure at the alpha promoters. Therefore, these results indicate that, in the experimental system used, HS-40 behaves as a strong inducible enhancer of human alpha- globin gene expression.     

Cytochrome P450 2B6 516G→T is associated with plasma concentrations of nevirapine at both 200 mg twice daily and 400 mg once daily in an ethnically diverse population

Objectives

The aim of the study was to characterize the impact of the cytochrome P450 2B6 (CYP2B6), CYP3A4, CYP3A5 and ATP‐binding cassette sub‐family B member 1 (ABCB1) polymorphisms on nevirapine plasma concentrations.

Methods

A total of 104 patients (82% male; 26% non‐Caucasian) were genotyped for eight single nucleotide polymorphisms at four loci (CYP2B6, CYP3A4, CYP3A5 and MDR1). Nevirapine plasma concentrations were determined using high‐performance liquid chromatography.

Results

Non‐Caucasian ethnicity [5609 ng/mL (n=27) for non‐Caucasians vs. 3771 ng/mL (n=77) for Caucasians; P<0.0001] and CYP2B6 516G→T [GG, 3574 ng/mL (n=50); GT, 4634 ng/mL (n=50); TT, 8170 ng/mL (n=4); P_{analysis of variance (anova)} =0.001] were significantly associated with a higher nevirapine trough concentration (C_trough). The latter association was maintained with both 200 mg twice daily (bid) and 400 mg once daily (qd) dosing [GG, 3527 ng/mL (n=30); GT, 4525 ng/mL (n=32); TT, 7020 ng/mL (n=2); P_anova =0.05 and GG, 3645 ng/mL (n=20); GT, 4861 ng/mL (n=17); TT, 9508 ng/mL (n=2); P_anova =0.01, respectively]. In a multivariable analysis, CYP2B6 516G→T and non‐Caucasian ethnicity remained significant predictors of nevirapine C_trough but CYP2B6 516G→T homozygosity had the greatest effect (108% higher, 46% higher). No associations were found between nevirapine C_trough and the remaining polymorphisms.

Conclusion

In this population, both non‐Caucasian ethnicity and carriage of the variant allele of CYP2B6 516G→T were significant predictors of nevirapine C_trough. The association between CYP2B6 516G→T and higher plasma nevirapine exposure was maintained at both bid and qd dosing.     

A K-ras oncogene increases resistance to sulindac-induced apoptosis in rat enterocytes

BACKGROUND & AIMS: Mutations of c-K-ras occur commonly in colonic neoplasms. The aim of this study was to determine how c-K-ras mutations alter the responses to the chemopreventive agent sulindac. METHODS: The parental rat intestinal cell line IEC-18 and c-K-ras-transformed derivatives were treated with sulindac sulfide. Cell cycle distribution was determined by flow-cytometric analysis (fluorescence-activated cell sorter), apoptosis by DNA fragmentation (laddering), flow cytometry, and microscopy, and changes in gene expression by immunoblotting. RESULTS: Sulindac sulfide inhibited cell growth and induced apoptosis in a time- and dose-dependent manner more rapidly in and at lower concentrations in parental cells than ras-transformed cells. Expression of the sulindac sulfide arrested cells in G0/G1, but cells entered apoptosis throughout the cell cycle. Proapoptotic protein Bak was relatively high in untreated parental cells and increased markedly after sulindac sulfide but was low in untreated ras-transformed cells and did not increase after sulindac sulfide. Expression of other Bcl-2 family members was unchanged after sulindac sulfide. However, sulindac sulfide reduced levels of cyclin D1 protein and cyclin E- and cyclin D1- associated kinase activity. CONCLUSIONS: c-K-ras-transformed enterocytes are relatively resistant to sulindac sulfide-induced growth inhibition and apoptosis, which may result from specific reduction of bak expression. (Gastroenterology 1997 Dec;113(6):1892-900)     

Trisomy 21 in childhood acute lymphoblastic leukemia: a Pediatric Oncology Group study (8602)

Of 1,036 children with newly diagnosed non-T, non-B acute lymphoblastic leukemia (ALL) and a demonstrated cytogenetic abnormality treated on the frontline Pediatric Oncology Group (POG) therapeutic trial 8602, there were 33 patients with trisomy 21 as the sole abnormality. Of these 33, 14 had Down syndrome (DS). Although the non-DS (NDS) trisomy 21 cases tended to be older than the DS cases, there were no other significant differences in clinicobiologic features nor in treatment outcomes between the DS and NDS groups, nor between the entire trisomy 21 group and the other chromosome abnormality group. Among NDS patients with +21 and one additional abnormality, +X, +16, -20, and structural abnormalities involving 6q or 12p were common findings. Kaplan-Meier event-free survival (EFS) curves showed a 4-year EFS of 80% (SE, 12%) in NDS trisomy 21 cases, 71% (SE, 22%) in DS cases with trisomy 21 as the sole abnormality, and 69% (SE, 2%) in cases with other chromosome abnormalities. Trisomy 21 as a sole acquired abnormality in NDS patients suggests a good prognosis.     

Effects of recombinant interleukin-2 administration on cytotoxic function following high-dose chemo-radiotherapy for hematological malignancy

Activated killer cells, unrestricted by major histocompatibility (MHC) antigens circulate in the peripheral blood of patients who have undergone autologous and allogeneic bone marrow transplant (BMT) and may contribute to the reduced risk of leukemic relapse observed after these procedures. Interleukin-2 (IL-2) in vitro augments this cytotoxicity and used therapeutically might thereby promote the eradication of minimal residual disease. In order to assess whether these effects on cytotoxicity can be reproduced in vivo, we studied changes in number, phenotype, and MHC unrestricted cytotoxicity of peripheral blood mononuclear cells obtained from patients with hematologic malignancy receiving IL-2 infusions. Patients with acute myeloid leukemia and multiple myeloma were treated after cytotoxic chemotherapy or autologous BMT. IL-2 infusions produced an initial lymphopenia, followed by a progressive recovery in mononuclear cell numbers and a rebound lymphocytosis after the termination of treatment. This affected all lymphocyte subsets; in particular CD25 (IL-2 receptor) positive cell numbers rose sevenfold. Cells with the ability to kill a natural killer (NK)-resistant, lymphokine activated killer cell (LAK)-sensitive target appeared in the circulation during 16 of 19 infusions and mean LAK activity rose from 5.9% to 15.5% during infusion (E:T ratio, 50:1; P less than .001). During IL-2 infusion, cells present in the peripheral blood inhibited the growth of myeloid leukemia blasts in agar after overnight co-culture. Depletion experiments showed that LAK activity was mediated by cells of both CD3- CD16+ (NK derived) and CD3+ CD16- (T derived) subsets. LAK precursor activity in peripheral blood also significantly increased during IL-2 infusion. Increases in major histocompatibility complex (MHC) unrestricted cytotoxicity can be produced by IL-2 infusions in vivo and may result in improved relapse-free survival following chemotherapy or BMT.