Genetic Analysis of Rapid Tolerance to Ethanol's Incoordinating Effects in Mice: Inbred Strains and Artificial Selection

Ethanol tolerance, a decrease in drug responsiveness with repeated administrations, is an important diagnostic criterion for alcoholism. Rapid tolerance develops within 8-24 hours of an initial ethanol exposure and shares many similarities with chronic tolerance. The genetic contribution to rapid tolerance to ethanol-induced ataxia was estimated using a panel of inbred strains of mice. Strains differed significantly in the degree of rapid tolerance development, which had a broad-sense heritability estimate of 0.11. Artificial selection was carried out to develop lines of mice that would show High (HRT) and Low (LRT) levels of Rapid Tolerance. Starting with HS/Npt mice, derived from a systematic cross of eight inbred strains, a significant response to selection was seen in replicate 1 by the third selection generation. No difference was found in replicate 2. Heritability estimates after the fourth generation were 0.25 for HRT-1 mice and 0.06 for LRT-1 mice. HRT-1 and LRT-1 mice also differed significantly in chronic tolerance development to four doses of ethanol. These studies provide evidence for a genetic contribution to rapid tolerance and support a genetic link between rapid and chronic tolerance to ethanol's ataxic effects.     

雌激素处理DCs在诱导小鼠同种皮肤移植免疫耐受中的作用

目的：研究雌二醇（17β-estradiol, E₂）处理的供体骨髓来源树突细胞（dendritic cells, DCs）在诱导小鼠同种异基因皮肤移植免疫耐受中的作用。方法：体外培养小鼠骨髓来源DCs，然后用E₂处理。受体小鼠在皮肤移植前经尾静脉分别输注经E₂处理的供体小鼠DCs、供体小鼠成熟DCs以及不成熟DCs，输注PBS为对照组。1周后对受体小鼠进行同种异基因皮肤移植，手术后观察皮肤存活情况，应用流式细胞术检测手术前后受体小鼠外周血中CD4+CD25+调节性T细胞百分率的变化。结果：在同种异基因皮肤移植中，受体小鼠尾静脉输注经E2处理的DCs组与对照组和不成熟DCs组比较，移植皮肤存活时间显著延长，较不成熟DCs组存活时间平均延长10.6 d（P<0.01）；与对照组和不成熟DCs组比较，E₂处理组外周血中CD4+CD25+ 调节性T细胞百分率明显升高（P<0.01）。结论：在同种异基因小鼠皮肤移植模型中，E₂处理的供体小鼠DCs可以延长移植皮肤存活时间。     

供体骨髓细胞输注促进小鼠皮肤移植耐受及其机理的研究

目的　探索“细胞 +环磷酰胺 (cyclophosphamide ,CP)”系统联合抗H 2 b 单克隆抗体、供体骨髓细胞输注诱导移植耐受的作用及其机制。方法　第 0天 ,经尾静脉给C5 7BL/ 6 (H 2 b,B6 )小鼠注入 10 8BALB/c(H 2 d,B/c)来源的脾细胞 ,第 2天 ,腹腔注射环磷酰胺 2 0 0mg/kg ,第 3、5天 ,分别经尾静脉注入抗H 2 b 单抗 ,剂量为 40 0 μg/ 0 .5ml,第 8天进行皮肤移植。皮肤移植后 1周 (第 15天 ) ,输入 2× 10 7供体BALB/c来源的骨髓细胞。观察皮肤移植物存活时间 ,并于第 30天对耐受B6小鼠作混合淋巴细胞反应 (mixedlymphocytereaction ,MLR) ,迟发型超敏反应 (delayedtypehypersensitivity ,DTH)等确定耐受的状态。并通过过继转移实验、嵌合体检查及脾细胞中细胞因子mRNA的表达情况 ,进一步探讨耐受形成的机制。结果　采用此诱导方案 ,B6小鼠对BALB/c小鼠的皮肤移植物长期存活。耐受可以被过继转移 ;耐受小鼠胸腺内的嵌合程度与耐受的维持密切相关 ;TH1型细胞因子在耐受小鼠中明显降低 ,而TH2型细胞因子明显升高。结论　“细胞 +CP”系统联合H 2 b 单克隆抗体、供体骨髓细胞输注能够诱导异基因小鼠皮肤移植耐受。耐受机制与胸腺嵌合体的存在密切相关。克隆无能 (aner gy)、抑制细胞和TH1/TH2偏移在耐受中也     

Fas在妊娠过程中表达变化的研究

目的探讨Fas在正常妊娠过程及妊娠期高血压疾病(HDCP)中的表达变化及意义。方法分别选取正常妊娠早期和晚期及HDCP胎盘绒毛组织。采用免疫组织化学方法检测Fas表达,并用高清晰彩色病理图像分析系统对其表达进行分析。结果Fas在妊娠早期胎盘组织中极少表达,妊娠晚期胎盘组织中表达增高。HDCP胎盘组织中Fas表达明显升高,与前两组比较差异有极显著性(P<0.01)。结论孕早期滋养细胞表面Fas表达在母胎免疫耐受中发挥重要作用,孕晚期Fas表达增加可能是引起HDCP的原因之一。     

Strain differences in mice in the development of tolerance to the anti-pentylenetetrazole effects of diazepam

The development of tolerance to the protective effects of diazepam (4 mg/kg) against seizures induced by pentylenetetrazole (PTZ) were studied in 3 strains of mice. Significant tolerance developed to protection against myoclonic jerks induced by PTZ (90-100 mg/kg) by day 5 in Tuck No. 1 and by day 10 in C3H/HE and CD-1 mice. Tolerance developed to protection against tonic-clonic convulsions by day 10 in Tuck No. 1 mice and by day 30 in the other strains. Diazepam remained protective against tonic-clonic convulsions (but not against myoclonus) induced by threshold doses of PTZ for 30 days in all 3 strains.     

An open,randomized single-centre study to compare the efficacy and convenience of follitropin beta administered by a pen device with follitropin alpha administered by a conventional syringe in women undergoing ovarian stimulation for IVF/ICSI

BACKGROUND: A pen device, similar to an insulin pen, has been recently marketed for the administration of follitropin beta in cartridges. A randomized controlled trial was performed to compare the efficacy and convenience of this pen device delivering follitropin beta with a conventional syringe delivering follitropin alpha. METHODS: A total of 200 patients needing IVF/ICSI treatment and willing to self-inject were enrolled in the study. All subjects had ovarian stimulation according to a long protocol and were randomized to the pen or the conventional syringe group during down-regulation by means of a computer-generated randomization list using random numbers. Patients were asked to fill in a daily local tolerance book after each injection. On the day of hCG the patients scored a Visual Analogue Scale (VAS) for pain and convenience. RESULTS: The average duration, total dose of recombinant FSH and number of cumulus oocyte complexes retrieved were 10.8/12.0 days (P = 0.001), 1880/2226 IU (P < 0.001) and 15.2/13.1 respectively in the pen device and conventional syringe groups; the presence of pain after the daily injection was significantly higher in the conventional syringe group (P = 0.027); the visual analogue scale score was similar for pain but significantly more convenient for the pen device (P < 0.001). The live birth rate per embryo transfer was 32.9 and 34.4% respectively in the pen device and conventional syringe groups. CONCLUSIONS: Self-injection with the pen device is safe and easy, more convenient and less painful for the patient, requires less FSH and shortens the treatment duration.     

Development of autoimmune insulitis is prevented in E alpha d but not in A beta k NOD transgenic mice

Two lines of E alpha d-expressing NOD mice were established by continuously backcrossing [E alpha d B6 transgenic mice x NOD] F1 to parental NOD or directly microinjecting the E alpha d gene into fertilized NOD eggs. Similarly, A beta k-expressing transgenic NOD mice were produced. Subsequent histological examination of pancreatic tissues revealed that autoimmune insulitis was prevented in E alpha d backcross and transgenic mice but not in A beta k transgenic mice.     

同种异体移植免疫耐受的进展

探索组织器官移植后的排斥反应,提高移植物的存活率是移植免疫研究者致力攻克的堡垒。20世纪90年代以来,随着器官移植术的广泛开展,人们对移植排斥反应的认识不断加深,对移植免疫耐受提出了新的研究思路和方法。目前普遍认为,解决异体移植排斥反应的关键在于诱导受体对供体的细胞、组织或器官产生免疫耐受,即受体免疫系统对移植物抗原产生特异性免疫无反应性。本文将对近年来阻断特异性免疫应答、诱导免疫偏离、主动免疫诱导同种移植耐受及建立受者体内嵌合体诱导移植耐受等主要方面的进展综述。     

DNA microarray technique for detection and identification of seven flaviviruses pathogenic for man

A flavivirus microarray was developed for detection and identification of yellow fever (YF), West Nile, Japanese encephalitis (JE), and the dengue 1-4 viruses, which are causing severe human disease all over the world. The microarray was based on 500-nucleotide probe fragments from five different parts of the seven viral genomes. A low-stringent amplification method targeting the corresponding regions of the viral genomic RNA was developed and combined with hybridization to the microarray for detection and identification. For distinction of the generated virus-specific fluorescence-patterns a fitting analysis procedure was adapted. The method was verified as functional for all seven flaviviruses and the strategy for the amplification, combined with the long probes, provided a high tolerance for smaller genetic variability, most suitable for these rapidly changing RNA viruses. A potentially high detection and identification capacity was proven on diverged strains of West Nile and dengue viruses. The lower limit for detection was equivalent, or better, when compared to routinely used RT-PCR methods. The performance of the method was verified on human patient samples containing dengue viruses, or normal human serum spiked with YF or JE viruses. The results demonstrated the ability of the flavivirus microarray to screen simultaneously a sample for several viruses in parallel, in combination with a good lower limit of detection.