Effect of tramadol on immune responses and nociceptive thresholds in a rat model of incisional pain

Objective： To evaluate the effects oftramadol on the proinflammatory responses in a rat model of incisional pain by investigating its effects on nociceptive thresholds and serum interleukin-6 （IL-6） and IL-2 levels. Methods： Forty-two male Sprague-Dawley （SD） rats scheduled for plantar incision were randomly divided into 7 groups 01=6 in each group）. Rats in Group 1 receiving general anesthesia with no incision were served as control; At 30 min before skin incision, Groups 2-5 were given 5 ml normal saline or 1, 10, and 20 mg/kg tramadol, respectively, intraperitoneally （i.p.）; Group 6 received 10 mg/kg tramadol after operation; Group 7 received 10 mg/kg tramadol before incision, followed by 200 μg/kg naloxone after operation. Mechanical allodynia was measured by electronic yon Frey filament to evaluate the nociceptive thresholds 1 h before incision, and 1 h and 2 h after operation. Serum IL-6 and IL-2 levels were measured by enzyme-linked immunosorbent assay （ELISA） 2 h after operation. Results： Mechanical thresholds decreased significantly and serum IL-6 level increased significantly after operation in Group 2 compared with control （P〈0.01）, and these changes were reversed respectively by tramadol in a dose-dependent manner （P〈0.05 and P〈0.01, respectively）. IL-2 level remained unchanged after operation in Group 2, but decreased in Group 3 （P〈0.05）, then gradually returned to the normal level in Groups 4 and 5. The intraperitoneally injected tramadol （10 and 20 mg/kg） produced a potent and dose-dependent antinocicptive effect on the lesioned paw. The antinocicptive effects of tramadol were partially an- tagonized by naloxone （200 μg/kg）, suggesting an additional non-opioid mechanism. Conclusion： The results suggest that tramadol could be a good choice for the treatment of pain under the conditions that immunosuppression may be particularly contraindicated.     

神经网络模型对造血干细胞体外扩增效果评价及预测

采用神经网络技术对造血干细胞HSCs体外扩增能力建立了评价及预测模型.总结了前人的实验结果,共获得341组数据.选取细胞接种密度、细胞因子组合、细胞来源、血清、基质细胞、反应器类型和培养时问等7个影响因子作为网络输入特征参数,分别对有核细胞(nUClear cells,NCs)、CD34 细胞和成集落细胞(colony-forming units,CFU-Cs)进行体外扩增能力的拟合评价及预测.选取124、90及86组实验数据分别用于NCs、CFU-Cs和CD34 细胞为评价指标的神经网络训练;而17、10及14组实验数据分别用于NCs、CFU-Cs和CD34 细胞为评价指标的神经网络预测.结果表明.对NCs、CFU-Cs和CD34 细胞的区间训练准确率分别为85.5%、86.7%和86.1%;区间预测准确率分别为82.4%、70.0%和71.4%.由此可知.人工神经网络的非线性模拟使定量描述HSCs体外扩增效果与培养条件问的关系及预测HSCs的最佳体外扩增条件成为可能.     

IL-6对大鼠脑缺血再灌注后海马及纹状体NOS和Fos表达的影响

目的 :研究IL 6对大鼠脑缺血再灌注 (CIRF)后海马及纹状体的NOS和Fos阳性神经元表达的作用。方法 :采用大脑中动脉栓塞法 (MCAO)对 2 4只Wistar大鼠左侧大脑中动脉构成CIRF模型的基础上分两组 :即单纯CIRF对照组 (n =8) ;经同侧脑室事先注入IL 6的实验组 (n =16)。分别进行NADPH脱氢酶反应显示NOS和免疫组织化学染色显示Fos表达。观察了CIRF后 1、2、4、6小时海马及纹状体处NOS和Fos阳性神经元数量的变化 ,以及IL 6对其变化的影响。结果 :大鼠CIRF后纹状体缺血中心区缺少NOS和Fos的表达 ,而纹状体周围区NOS和Fos阳性神经元明显增加 ;海马各部NOS和Fos阳性神经元均明显增加 ,其中CA1区增加最显著。但预先经侧脑室注射IL 6的实验组中 ,大鼠海马及纹状体NOS和Fos阳性神经元明显低于对照组。IL 6明显抑制了NOS及Fos表达。结论 :提示IL 6可能参与脑缺血性神经元损伤的调控作用     

谷氨酰胺对大强度跑台训练大鼠免疫功能影响的实验研究

目的:探讨补充谷氨酰胺对大强度跑台训练大鼠免疫功能的影响。方法:适应性训练后,30只6周龄健康雄性SD大鼠,随机分为安静组(C,6只),训练组(T,12只)和服药训练组(TG,12只)。进行6周大强度跑台训练,跑台坡度5°,前2周每周训练6d,每次训练25min,速度依次为20 m/min和24 m/min,后4周每天训练,速度和时间不变,分别为32 m/min和35min。训练期间每日给药前称体重,TG组大鼠按O.3g/kg.d剂量灌胃给药,C、T组喂以相同体重比例的自来水。最后一次训练12h后进行大鼠脾脏T淋巴细胞亚群CD4+、CD8+和脾细胞IL-2表达测定。结果:T组与C组比较,CD8+含量显著升高,CD4+/CD8+比值及IL-2表达显著下降;TG组与C组比较,CD8+含量显著下降,CD4+/CD8+及IL-2表达升高显著。结论:本该实验的大强度跑台训练造成大鼠免疫功能显著下降,而谷氨酰胺补充可明显改善大强度跑台训练造造成的大鼠免疫抑制,增强大鼠免疫功能。     

白介素24对单个核细胞bcl-2表达的影响

为探讨白细胞介素24(IL-24)对成人急性白血病骨髓单个核细胞(BMMNC)体外生长的影响,并初步研究其作用机制。随机选取急性白血病住院患者15例,均经FAB分型或免疫学分型确诊,其中确诊未治的成人急性淋巴细胞白血病(ALL)患者5例,成人急性非淋巴细胞白血病(ANLL)患者10例。取患者骨髓,分离单个核细胞并培养,将不同浓度IL-24作用于BMMNC。应用四氮唑蓝(MTT)比色还原法、AnnexinV-FITC染色流式细胞术及逆转录-聚合酶联反应(RTPCR),分别检测IL-24对急性白血病BMMNC体外细胞增殖与凋亡及bcl-2表达的影响。结果显示,IL-24对体外培养的白血病BMMNC有明显的生长抑制作用,且呈时间、剂量依赖性,与阴性对照组相比差异有统计学意义(P0.05)。IL-24浓度50 ng/mL作用急性白血病BMMNC,48 h后凋亡率为39.21%±5.79%,高于对照组(P0.01)。不同浓度IL-24处理白血病BMMNC,能够降低bcl-2 mRNA的表达。IL-24体外培养的成人急性白血病BMMNC,有明显的生长抑制、诱导凋亡作用。由此可知,IL-24可以下调bcl-2 mRNA表达,可能是IL-24抑制急性白血病BMMNC生长作用的重要机制。     

Investigation of gene expression profiles in coronary heart disease and functional analysis of target gene

The research outlined here includes constitution of the differential gene expression profile by means of oligonucleotide gene microarray and functional analysis of the target gene for coronary heart disease (CHD). In a microarray screening experiment, the predominance of inflammation- and immune-related genes is presented in the expression profile of 107 differential genes based on the analysis of gene ontology and gene pathway. IL-8, an inflammatory factor, is identified as one of the genes that were markedly up-regulated in CHD. The plasma level of IL-8 is significantly raised in patients with CHD (n = 30) compared with healthy controls (n = 40), which underscores the clinical relevance of the in vitro finding. The further functional analysis shows that IL-8 affects platelet aggregation percentage, expression of CD62p and platelet aggregation morphology in 12 healthy volunteers to some extent. These findings suggest the relevance of inflammation and immune responses to CHD at the DNA level. Moreover, IL-8 may be involved in the pathogenesis of CHD through the pathway of platelet activation. Supported by the National Natural Science Foundation of China (Grant No. 90409021)     

低功率毫米波辐射对HL60细胞基因表达谱的影响

摘要：研究低功率毫米波辐射对HL60白血病细胞基因表达谱的影响。应用基因芯片检测频率41.32GHz的毫米波辐射HL60白血病细胞和未辐射毫米波HL60白血病细胞组基因表达差异,并进行RT-PCR方法验证IL-7、EGF和LGALS3基因变化。 结果与对照组比较,毫米波辐射60min后,HL60细胞增殖,基因芯片检出基因表达上调18个和下调306个,在下调的基因中,RT-PCR 检出IL-7、EGF和LGALS3基因下调与基因芯片结果一致。表明低功率毫米波可导致HL60细胞基因表达谱发生变化,这些变化的基因与HL60细胞增殖功能相关。提示基因表达变化是低功率毫米波辐射HL60细胞所致生物学反应的重要因素。     

秋水仙素处理对黄花酢浆草生长的影响及诱变效应

以黄花酢浆草(Oxalis corniculata L.)腋芽为外植体,建立快速繁殖体系,并观察不同浓度秋水仙素处理对其生长的影响及诱变效应.结果表明:丛生芽诱导及增殖的最适培养基为MS+2.0mg.L-1 6-BA(6-苄氨基腺嘌呤)+1.5mg.L-1 NAA(α-萘乙酸),1/2MS+1.0mg.L-1 NAA生根效果最好.10mg.mL-1秋水仙素处理诱导丛生芽率达100%,且出现茎膨大、矮化等现象.     

升清降浊方防治慢性肾功能衰竭的临床研究

目的探讨升清降浊方治疗慢性肾功能衰竭的临床疗效。方法将60例患者随机分为升清降浊方组(治疗组)和包醛氧淀粉组(对照组)各30例。治疗组予升清降浊方中药治疗,水煎服,每日1剂。对照组予包醛氧淀粉,规格5g/包,口服,5g/次,3次/d。以上各组均以30 d为1疗程,连续治疗2个疗程。观察两组患者治疗前后的营养状态(血清白蛋白、血色素及体重等)、肾功能(血清肌酐(Scr)、尿素氮(Bun)、胱抑素C、尿蛋白等)及炎症因子指标(血清C反应蛋白、IL-1、IL-6、TNFa等)的变化。结果经过2个疗程60d的治疗并进行比较,治疗组和对照组CRF患者的上述指标均有极其显著差异(P 0.01)。结论升清降浊方治疗慢性肾功能衰竭疗效显著,对炎症因子的表达均有显著差异。     

RNA干扰沉默PID1基因在C2C12细胞中表达的研究

 构建和筛选对PID1（phosphotyrosine interaction domain containing 1, PID1）基因有RNA干扰作用的PID1-shRNA表达载体。据小鼠PID1 cDNA序列,优化设计了4条shRNA及1条阴性干扰序列,插入pGPU6/GFP/Neo载体中,得到pGPU6/GFP/Neo-PID1-1、pGPU6/GFP/Neo-PID1-2、pGPU6/GFP/Neo-PID1-3、pGPU6/GFP/Neo PID1 4和pGPU6/GFP/Neo-PID1-NC。干扰载体转染C2C12细胞,以RT-PCR和Western blot技术检测shRNA对C2C12细胞中PID1 mRNA和蛋白表达的下调作用。结果表明：靶向PID1基因的4个shRNA重组质粒载体经测序分析,其shRNA编码序列与预期设计的完全一致,经酶切鉴定和测序分析证实,靶向PID1基因的shRNA重组质粒载体构建成功。进一步将构建的4个表达载体分别转染C2C12细胞,24h后细胞中PID1基因mRNA表达水平依次下调 (23.58±1.87)%、(75.44±0.77)%、(70.52±0.41)% 和 (56.60±3.13)%。48 h后细胞中PID1蛋白表达水平依次降低 (30.15±5.05)%、(71.86±4.85)%、(67.93±2.28)% 和 (56.81±2.01)%。所筛选出的pGPU6/GFP/Neo-PID1-2、pGPU6/GFP/Neo-PID1-3和pGPU6/GFP/Neo-PID1-4三个表达载体均能高效地抑制转染细胞PID1 mRNA和蛋白的表达,为进一步研究PID1基因的功能奠定了基础。     

IL-12在哮喘动物模型BALF中的动态变化及意义

目的观察哮喘动物模型支气管肺泡灌洗液（branchoalveolar lavage fluids，BLAF）中白介素-12（interleukine-12，IL-12）的动态变化及对嗜酸性细胞（eosinophile，EOS）凋亡指数的影响。方法将62只Wistar大鼠喂养7d后，随机分为正常对照组（6只）、哮喘组（56只），哮喘组又随机各取8只分为7个不同的时相组。哮喘各时相组于第1天及第7天用蛋白粉（ovalbumin，OVA）10mg、氢氧化铝20mg和生理盐水制成的2ml悬液腹腔注射致敏，对照组用生理盐水2ml腹腔注射；哮喘各时相组第15d开始均用1％OVA悬液雾化吸入激发，1次／d，1h／次，共7d，正常对照组用生理盐水代替1％OVA悬液，余同哮喘组。末次激发后，对照组1h麻醉，哮喘组分别于1h、8h、24h、48h、96h,7d和14d麻醉，各组麻醉后行支气管肺泡灌洗（branchoalveolar lavage，BAL），用流式细胞仪测定BLAF中EOS凋亡指数，用酶联免疫吸附法（enzynle linked immunosorbent assay，ELISA）测定IL-12浓度。结果哮喘各时相组BALF中，IL-12浓度的动态变化呈双峰，1h上升，8～24h迅速回落到低峰，48h再次上升，96h达到高峰，此后下降，14d降至对照组水平；而EOS凋亡指数的动态变化呈双峰，1h上升，而8h下降，24h降至低峰，48～96h快速上升，达到高峰，随后下降，14d降至对照组水平；IL-12与EOS凋亡指数呈同步变化，两者作直线相关分析呈正相关。结论在哮喘大鼠气道中，IL-12可能通过调节EOS凋亡指数，从而调节气道的炎症反应。     

大鼠肝再生中肝细胞基因表达与DNA裸露的相关性研究

为了解大鼠肝再生中肝细胞的基因表达与DNA裸露的相关性,分离大鼠肝细胞,提取裸露DNA,用SOLEXA方法对DNA测序,用BWA软件拼装测序片段,用IPA软件分析拼装的基因种类和作用,用Rat Genome230 2.0芯片检测基因的转录情况.研究表明,大鼠肝再生的12h(PH后12h),肝细胞的裸露DNA涉及16 912个基因,1 701个基因发生了有意义裸露量变化.其中,25个基因的裸露量上调,1 676个基因的裸露量下调.与Rat Genome 230 2.0芯片的检测的基因转录比对表明,肝细胞的AKR7A3,BMYC,CDC25B等26个基因的裸露量和表达量均下调,表明大鼠肝再生12h时,这些基因的表达可能受染色体结构调控.     

Achyranthes bidentata Blume extract promotes neuronal growth in cultured embryonic rat hippocampal neurons

We have prepared an aqueous extract of Achyranthes bidentata Blume, a commonly prescribed Chinese medicinal herb, and reported, in previous studies, that A. bidentata extract benefits nerve growth and prevents neuron apoptosis. In this study, we investigated the actions of A. bidentata extract on survival and growth of primarily cultured rat hippocampal neurons. The morphological observation revealed that neurite growth from hippocampal neurons was significantly enhanced by A. bidentata extract with similar effects to those induced by nerve growth factor (NGF), and the greatest neurite growth appeared on treatment with A. bidentata extract at 1 μg/ml for 24 h. DNA microarray analysis indicated that there were 25 upregulated genes and 47 downregulated genes exhibiting significantly differential expression in hippocampal neurons treated with A. bidentata extract at 1 μg/ml for 6 h when compared to those in untreated hippocampal neurons. Real-time quantitative RT-PCR and Western blot analysis demonstrated that the expression of growth-associated protein-43 in hippocampal neurons was upregulated at both mRNA and protein levels after treatment with A. bidentata extract, and the optimal dosage of the extract was also 1 μg/ml. These data confirm that A. bidentata extract could promote in vitro hippocampal neuronal growth in a dose- and time-dependent manner.     

斯氏狸殖吸虫感染大鼠致肝纤维化形成过程中IL-4和IFN-γ表达及意义

 探讨斯氏狸殖吸虫感染大鼠致肝纤维化形成过程中细胞因子IL-4和IFN-γ表达及意义,为阐明Th1/Th2在斯氏狸殖吸虫感染大鼠肝纤维化形成及调节作用提供实验依据.SD大鼠36只,随机分为正常对照组(n=6)、感染1周组(n=5)、感染2周组(n=5)、感染4周组(n=5)、感染8周组(n=5)、感染12周组(n=5)和感染16周组(n=5),每鼠腹腔注射斯氏狸殖吸虫囊蚴15个.按感染时间处死各组大鼠,观测各项指标.HE染色和V G染色观察肝组织纤维化病理形态学变化.免疫组织化学法测定IL-4,IFN-γ在大鼠肝纤维化形成过程中的表达动态变化.HE染色和V G染色肝组织病理形态学结果显示:随着感染时间的延长,胶原纤维面积比值(S/T)和肝纤维化评级在逐渐增加,肝组织纤维化的程度逐渐加重.免疫组织化学显示:IFN-γ的表达阳性细胞百分率在1~8周逐渐增加,到第8周末时达到最大,在12周到16周又逐渐减少;而IL-4表达阳性细胞百分率在1~16周逐渐增加,到第12周末IL-4表达开始增加迅速. 斯氏狸殖吸虫感染大鼠可引起肝脏纤维化病变,IL-4和IFN-γ在斯氏狸殖吸虫感染大鼠致肝纤维化形成过程中表达增加,其纤维化病变程度与感染时间、IL-4和IFN-γ的表达增加有关.提示Th1细胞分泌的细胞因子IFN-γ和Th2细胞分泌的细胞因子IL-4在斯氏狸殖吸虫感染大鼠致肝纤维化的自发性免疫调节中起作用.     

红藻氨酸致癫痫大鼠不同时点神经生长因子变化研究

摘要: 目的 探讨红藻氨酸( Kainic Acid,KA) 致癫痫大鼠癫痫发作过程中海马组织神经生长因子( nerve growth factor,NGF) 不同时间点表达量的变化。方法 腹腔注射 KA 建立大鼠癫痫模型。采用 TaqMan 探针实时定量 PCＲ ( Ｒeal-time quantitative PCＲ) 技术,检测注射 KA 后不同时点大鼠海马组织中 NGF 表达量的变化。结果 与 NGF 表达量与生理盐水对照组( NS) 相比,6 ～ 12 h NGF 表达量明显低于 NS 组( P ＜ 0. 05) 、48 ～ 72 h NGF 表达量开始升高并高于 NS 组,在 24、72 h 时其表达量显著高于 NS 组( P ＜ 0. 05) 。结论 NGF 对致癫大鼠的海马具有修复与保护作用。     

H-1 细小病毒感染大鼠神经胶质瘤细胞C6 机制的初步探讨

摘要: 目的探讨H-1 细小病毒感染大鼠神经胶质瘤细胞C6 后细胞因子水平的变化。方法取感染C6 细胞后不同时间点的H-1 细小病毒,同时取相同时间点的正常C6 细胞,采用ELISA 方法检测同时检测TNFα、IL-6、TGF-β1和IL-17A 的表达水平。结果正常C6 细胞中TNF-α、TGF-β1 和IL-17A 的浓度随培养时间增长而提高, IL-6 浓度无明显变化; H-1 细小病毒感染的C6 细胞中TNF-α、TGF-β1 和IL-17A 浓度无明显变化,而IL-6 浓度随感染时间推移而显著增加。结论H-1 细小病毒感染导致C6 细胞中细胞因子表达水平的变化。     

A role for Wnt signalling in self-renewal of haematopoietic stem cells

Haematopoietic stem cells (HSCs) have the ability to renew themselves and to give rise to all lineages of the blood; however, the signals that regulate HSC self-renewal remain unclear. Here we show that the Wnt signalling pathway has an important role in this process. Overexpression of activated beta-catenin expands the pool of HSCs in long-term cultures by both phenotype and function. Furthermore, HSCs in their normal microenvironment activate a LEF-1/TCF reporter, which indicates that HCSs respond to Wnt signalling in vivo. To demonstrate the physiological significance of this pathway for HSC proliferation we show that the ectopic expression of axin or a frizzled ligand-binding domain, inhibitors of the Wnt signalling pathway, leads to inhibition of HSC growth in vitro and reduced reconstitution in vivo. Furthermore, activation of Wnt signalling in HSCs induces increased expression of HoxB4 and Notch1, genes previously implicated in self-renewal of HSCs. We conclude that the Wnt signalling pathway is critical for normal HSC homeostasis in vitro and in vivo, and provide insight into a potential molecular hierarchy of regulation of HSC development.