抑制TNFR/RIPK信号通路对大鼠急性脊髓损伤后神经功能的影响

目的 :探讨应用坏死性凋亡抑制剂Necrostatin-1(Nec-1)抑制TNFR/RIPK介导的坏死性凋亡信号通路对大鼠急性脊髓损伤(spinal cord injury,SCI)的作用。方法:72只雄性SPF级SD大鼠,体重0.25~0.30kg,随机分为4组:假手术组(Sham组,A组)、假手术+Necrostatin-1组(Sham+Nec-1组,B组)、SCI+二甲基亚砜(DMSO)(DMSO组,C组)、SCI+Nec-1组(Nec-1组,D组)。C、D组采用钳夹法制作大鼠急性SCI模型。所有大鼠硬膜下置管,A组不给药,B组和D组造模后30min经导管注射1μl Nec-1(25μg/μl),C组注射等量DMSO,1次/d,至取材时间点。各组分别于造模后12h、24h和3d三个时间点每组取6只大鼠,先行Basso/Beattie/Bresnahan(BBB)评分,再处死动物取脊髓组织。12h时间点动物处死前1h腹腔注射碘化丙啶(propidine iodide,PI)1mg/kg,取材检测脊髓组织PI红染细胞数;24h时取材采用苏木素-伊红(HE)染色观察脊髓损伤情况、尼氏(Nissl)染色观察神经元存活数目、Western Blot(WB)检测Bcl-2、坏死性凋亡蛋白RIPK1及RIPK3的表达水平;3d时取材行Tunel染色观察细胞凋亡情况。结果:造模后A组和B组各时间点的BBB评分均正常,C、D组各时间点均显著性低于A、B组,D组各时间点的BBB评分均显著高于同时间点C组(P0.05)。造模后12h,D组PI红染细胞较C组明显减少,神经元崩解减轻(P0.05)。造模后24h,A组和B组脊髓组织HE和Nissl染色正常,D组脊髓组织损伤程度和存活神经元数量均优于C组,差异有统计学意义(P0.05);A组和B组Bcl-2、RIPK1及RIPK3均低水平表达,C组RIPK1及RIPK3表达显著性升高,D组Bcl-2表达较C组上调,RIPK1及RIPK3表达显著性下降,C、D两组比较差异均有统计学意义(P0.05)。造模后3d,A组和B组可见少量凋亡小体,C组和D组明显增多,但D组凋亡小体数量较C组明显减少(P0.05)。结论:抑制TNFR/RIPK信号通路可以减轻大鼠急性SCI后的病理变化,改善行为学评分,促进脊髓神经功能恢复。     

不同时间点应用甲氨蝶呤对大鼠脊髓损伤后神经细胞凋亡的影响

目的探讨不同时间点应用低剂量甲氨蝶呤(methotrexate,MTX)对大鼠脊髓损伤(spinal cord injury,SCI)后神经细胞凋亡的作用,探讨其潜在的神经保护机制与合适的给药时机。方法取成年雄性SD大鼠120只,体质量247~286 g,随机分为4组(n=30),分别为假手术组(A组)、对照组(B组)、MTX治疗组(C组)和MTX预防组(D组)。A组仅行椎板切除,B、C、D组采用改良Allen法制备SCI模型;C组于术后1、6、12、18、24 h,D组于术前30 min及术后6、12、18、24 h,经尾静脉注射MTX(0.5 mg/kg);A、B组于术后1、6、12、18、24 h注射等量生理盐水。术后观察大鼠一般情况,于1、3、7、14、21 d采用BBB评分进行神经功能评估;取材行组织学观察脊髓形态学变化,免疫组织化学染色观察半胱氨酸天冬氨酸蛋白酶3(Caspase-3)表达,TUNEL标记观察细胞凋亡水平。结果 B、C、D组实验过程中共死亡10只大鼠,最终各组纳入25只大鼠进行观察。各时间点A组BBB评分均高于B、C、D组(P0.05),从3 d开始C、D组评分明显高于B组(P0.05);D组从3 d开始均高于C组,除21 d(P0.05)外,其余各时间点比较差异均有统计学意义(P0.05)。组织学观察示,A组术后各时间点脊髓组织为正常结构;D组SCI程度轻于B、C组,C组轻于B组;从14 d开始,B、C、D组病变范围基本固定。各时间点B、C、D组Caspase-3及TUNEL阳性细胞数均显著多于A组,B组多于C、D组,比较差异有统计学意义(P0.05);C组Caspase-3阳性细胞数均多于D组,其中3、7、14 d比较差异有统计学意义(P0.05),而TUNEL阳性细胞数仅3、7 d显著多于D组(P0.05)。术后1、3、7、14、21 d,B、C、D组组内Caspase-3、TUNEL阳性细胞数均成正相关(P0.05)。结论低剂量MTX能够通过抑制神经细胞凋亡有效阻止SCI的继发性损伤;预防性应用MTX效果优于单纯治疗性应用。     

体外转基因成肌细胞移植对大鼠损伤脊髓细胞凋亡的影响

目的：探讨大鼠脊髓损伤后胚胎脊髓和腺病毒介导的脑源性神经生长因子（AxCA-BDNF)体外转基因成肌细胞移植对大鼠脊髓细胞凋亡的影响。方法：将动物分为：大鼠脊髓半切洞损伤明胶海绵填充组（A组）,大鼠脊髓半切洞损伤应用胚胎脊髓移植组（B组）,脊髓半切洞损伤损伤AxCA-BDNF基因转染的成肌细胞移植组（C组）大鼠脊髓半切洞损伤后应用胚胎脊髓和AxCA-BDNF基因转染的成肌细胞移植组（D组）。手术后1、3、7、14、28d应用行为学和电生理检查观察大鼠功能恢复情况,对脊髓损伤区进行细胞凋亡的检测（TUNEL）以及Bcl-2蛋白表达的测定（免疫组化法）。采用计算机图像分析技术,进行定量分析。结果：A、B、C、D四组中均发现凋亡细胞及Bcl-2蛋白阳性表达细胞,图像分析发现,各组凋亡细胞核为A＞B＞C＞D;Bcl-2免疫反应阳性细胞表达顺序为D＞C＞B＞A,Bcl-2免疫反应阳性细胞的表达与大鼠后肢功能恢复有同样的变化趋势。结论：大鼠胚胎脊髓和体外转基因成肌细胞移植能抑制脊髓损伤后的细胞凋亡。     

芒果苷对大鼠急性脊髓损伤的保护作用及相关机制研究

目的探讨芒果苷对大鼠急性脊髓损伤(spinal cord injury,SCI)的保护作用并分析其相关机制。方法成年雄性SD大鼠90只,体质量250~300 g,随机分为假手术组(A组)、SCI组(B组)、10 mg/kg芒果苷处理组(C组)、25 mg/kg芒果苷处理组(D组)、50 mg/kg芒果苷处理组(E组),每组18只。B、C、D、E组采用Allen法(60 g/cm)建立大鼠T9 SCI模型,A组仅切除T8~10椎板;C、D、E组术后每天按照10、25、50 mg/kg剂量腹腔注射芒果苷,共注射30 d;A、B组于对应时间点注射等量生理盐水。术后观察大鼠存活情况,于24、48、72 h采用BBB评分评价大鼠后肢运动功能;72 h时取损伤节段脊髓测量其水含量,ELISA检测氧化应激反应因子丙二醛(malondialdehyde,MDA)、过氧化氢酶(catalase,CAT)、过氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH)以及炎性因子NF-κB、TNF-α、IL-1β、IL-6活性;30 d时取损伤节段脊髓采用ELISA法检测Caspase-3、9活性,Western blot检测凋亡蛋白Bax、Bcl-2表达,组织学观察脊髓组织形态,免疫组织化学染色观察Caspase-3蛋白表达。结果术后各组大鼠均存活至实验结束。B、C、D、E组BBB评分均显著低于A组,B、C、D、E组评分呈逐渐增高趋势,组间比较差异均有统计学意义(P0.05)。B、C、D、E组脊髓水含量均显著高于A组,B、C、D、E组水含量呈逐渐降低趋势,组间比较差异有统计学意义(P0.05)。ELISA检测示,B、C、D、E组MDA、NF-κB、TNF-α、IL-1β、IL-6、Caspase-3、Caspase-9活性均显著高于A组,B、C、D、E组均呈逐渐降低趋势;而CAT、SOD、GSH活性均显著低于A组,B、C、D、E组均呈逐渐增加趋势;以上指标组间比较差异均有统计学意义(P0.05)。Western blot示,B、C、D、E组Bax蛋白表达明显高于A组,B、C、D、E组表达呈逐渐降低趋势;Bcl-2蛋白表达明显低于A组,B、C、D、E组表达呈逐渐增高趋势;组间比较差异有统计学意义(P0.05)。组织学观察示B组脊髓组织符合SCI病理改变,C、D、E组神经坏死程度较B组好转,并且E组效果优于D组,D组优于C组。免疫组织化学染色观察,B、C、D、E组Caspase-3蛋白表达量显著高于A组,B、C、D、E组呈逐渐降低趋势(P0.05)。结论对于大鼠急性SCI,芒果苷可通过减轻脊髓组织水肿、抑制氧化应激反应及炎性反应,调节Bax、Bcl-2蛋白表达,从而发挥神经保护作用。     

银杏叶提取物对大鼠急性脊髓损伤后神经功能恢复的作用及其机制

目的:观察银杏叶提取物(EGb)对大鼠实验性脊髓损伤后组织结构及运动功能恢复的作用,探讨其对急性脊髓损伤的作用机制。方法:120只SD雄性大鼠,随机分为假手术组(A组)、损伤对照组(B组)、甲基强的松龙(MP)治疗组(C组)和EGb治疗组(D组),每组30只。B、C、D组用Allen′s法以50g·cm致伤大鼠T9脊髓制作损伤模型,B组为单纯脊髓损伤,不给药;C组为脊髓损伤后30min内,由腹腔注入MP30mg/kg;D组为术后至处死前每天腹腔给予EGb17.5mg/kg;A组只打开T9椎板,不打击脊髓,不给药。术后24h、3d、5d、7d、14d对大鼠进行脊髓运动功能(BBB)评分。于术后24h、3d、5d、7d、14d处死动物(n=6),取T9节段脊髓,切片苏木素-伊红(HE)染色观察脊髓大体组织结构变化,用免疫组织化学方法检测B细胞淋巴瘤/白血病基因-2(Bcl-2)和B细胞淋巴瘤/白血病基因伴随蛋白x(Bax)在脊髓前角运动神经元中的表达变化情况。结果:各时间点B、C、D组大鼠脊髓运动功能(BBB)评分均显著低于A组(P<0.01),伤后7d、14d时C、D组评分显著高于B组(P<0.05),各时间点C组与D组评分比较无统计学意义(P>0.05)。HE染色C组和D组大鼠脊髓损伤区较B组坏死程度轻、形成囊腔少,A组正常;1周后D组较C组片状出血灶少,神经细胞肿胀不明显。各时间点B、C、D组大鼠损伤脊髓前角运动神经元中Bcl-2阳性细胞数均显著高于A组(P<0.01),C、D组显著高于B组(P<0.01),7d、14d时D组显著高于C组(P<0.05);各时间点B、C、D组大鼠损伤脊髓前角运动神经元中Bax阳性细胞数均显著高于A组(P<0.01),C、D组显著低于B组(P<0.01),7d、14d时D组显著低于C组(P<0.01)。结论:EGb可能通过抑制Bax表达、提高Bcl-2表达,抑制脊髓损伤后神经元凋亡,在运动功能恢复、损伤脊髓组织保护上发挥其有益作用,1周后EGb仍能抑制脊髓损害后的继发性损伤。     

脊髓减压联合电针对急性上颈段重度脊髓压迫损伤影响的实验研究

目的:探究脊髓减压联合电针对急性上颈段重度脊髓压迫损伤大鼠的治疗效果和可能的作用机制。方法:SPF级Wistar大鼠30只随机分为5组(对照组A、B,实验组C、D、E),每组6只,采用经寰枕间隙置入球囊导管加压造成脊髓压迫损伤的方法构建脊髓损伤模型。A组无任何干预,空白对照,B组置入球囊导管后不加压,做假手术对照,C、D、E组加压48 h后松球囊脊髓减压,C组取百会、大椎穴进行电针干预,连续波,频率2 Hz,治疗时间15 min,持续治疗14 d,D组于大鼠尾静脉注射甲强龙进行甲强龙冲击治疗,E组不再行任何治疗干预。14 d后5组大鼠全部行腹主动脉取血和脊髓损伤组织取材,采用BBB评分法对5组大鼠的运动功能进行评价,ELISA法检测损伤组织及血清中血小板活化因子(platele-activating factor,PAF)的含量,Western blot法检测损伤组织中Caspase-9的表达。结果:BBB评分:对照组A、B在实验组压迫后1、48 h,实验组治疗后24 h,3、7、14 d等6个时间点上均为(21.000±0.000)分,实验组评分始终低于对照组,C、D组评分显著高于E组(P0.05),C、D组评分相近(P0.05)。ELISA法测PAF结果显示:A、B、D、E组血清中PAF浓度相近(P0.05),C组血清中PAF浓度较其余4组低(P0.05),A、B组组织中PAF浓度结果相近(P0.05),C组组织中PAF浓度较A、B组高(P0.05),D组组织中PAF浓度较A、B、C组高(P0.05),E组组织中PAF浓度较其余4组高(P0.05);Western blot法检测结果显示:A、B组Caspase-9的表达量相近(P0.05),C组表达较A、B组高(P0.05),D组表达量较A、B、C组高(P0.05),E组表达较A、B、C、D组高(P0.05)。结论:脊髓减压联合电针治疗急性上颈段重度脊髓压迫损伤较脊髓减压联合甲强龙和单纯脊髓减压效果更佳,其作用机制可能与降低脊髓损伤组织中PAF的含量与下调其Caspase-9蛋白的表达有关。     

表达VEGF的重组腺相关病毒对大鼠创伤性脊髓损伤的保护作用及其机制研究

目的探讨表达VEGF的重组腺相关病毒(recombinant adeno-associated virus,rAAV)对大鼠创伤性脊髓损伤(spinal cord injury,SCI)的保护作用及其机制。方法取144只成年雄性SD大鼠(体重200～250 g)随机分为4组,每组36只。假手术组(A组)仅手术暴露脊髓。模型对照组(B组)、rAAV-绿色荧光蛋白(green fluorescentprotein,GFP)组(C组)、rAAV-hVEGF165-GFP组(D组)制备创伤性SCI大鼠模型,术后即刻分别予以20μL生理盐水、rAAV-GFP病毒、rAAV-hVEGF165-GFP病毒脊髓注射治疗。术后3、7 d采用BBB评分观察大鼠后肢运动功能变化;术后7 d通过脊髓组织尼氏小体染色观察脊髓组织病理学变化,脊髓组织神经元透射电镜检测及TUNEL凋亡细胞染色观察脊髓神经元凋亡,Western blot检测水通道蛋白4(aquaporin 4,AQP-4)表达;术后1、3、5、7 d ELISA法检测VEGF165蛋白表达。结果 BBB评分显示术后3、7 d D组神经功能较B、C组显著改善(P<0.05)。尼氏小体染色显示术后7 d D组脊髓组织结构破坏明显轻于B、C组(P<0.05)。ELISA检测显示D组VEGF165蛋白表达呈低剂量缓释表达,术后3、5、7 d,D组VEGF165蛋白表达均高于其他3组(P<0.05)。透射电镜检测及TUNEL凋亡细胞染色结果显示术后7 d D组脊髓神经元凋亡较B、C组显著减少,凋亡率显著低于B、C组(P<0.05)。Western blot结果显示术后7 d D组脊髓组织AQP-4蛋白表达明显较B、C组减少(P<0.05)。结论表达VEGF的rAAV通过抗神经元凋亡及减轻脊髓水肿对大鼠创伤性SCI起保护作用。     

罗西格列酮对脊髓损伤大鼠神经功能恢复的作用及机制

目的:观察罗西格列酮对脊髓损伤(SCI)大鼠后肢运动功能恢复的作用,探讨其作用机制。方法:75只成年SD大鼠,应用Allen改良法制作大鼠T10SCI模型,随机分为A、B、C三组,每组25只,B、C组于损伤后5min、6h、24h腹腔注射罗西格列酮,C组在腹腔注射罗西格列酮前1h给予G3335,A组于相应时间点腹腔注射等体积生理盐水作为对照组。每组取6只大鼠于伤后1d、7d、2w、4w、6w时对后肢运动功能进行BBB评分;伤后3d每组取4只动物脊髓组织行免疫组织化学染色法检测核转录因子κB(nuclear factor kappa-light-chain-enhancer of activated B cells,NF-κB)的表达;伤后1、3、5、7d和2w每组取3只应用Westernblot法检测脊髓组织中凋亡相关蛋白caspase-3和Bcl-2的表达。结果:伤后1d、7d时3组大鼠BBB评分均为0分,伤后2w开始B组BBB评分高于A组和C组,4w和6w时与A、C组比较有显著性差异(P0.05);伤后3d时三组NF-κB表达均为阳性,但B组平均光密度值明显低于A、C组(P0.05),B组与C组比较无显著性差异(P0.05);伤后各时间点B组caspase-3表达量均低于A组和C组(P0.05),而Bcl-2表达均高于A组和C组(P0.05),其差异均在伤后5d达到高峰,A组与C组同时间点比较无显著性差异(P0.05)。结论:罗西格列酮可促进SCI大鼠神经功能恢复,其机制可能与抑制炎症反应及细胞凋亡有关。     

N-甲基-D-天门冬氨酸受体拮抗剂MK-801胚胎脊髓移植后大鼠损伤脊髓凋亡的影响

[目的]探讨大鼠脊髓损伤后N-甲基-D-天门冬氨酸(NMDA)受体拮抗剂MK-801胚胎脊髓移植后对大鼠脊髓细胞凋亡的影响.[方法]将动物分为大鼠脊髓半切洞损伤后,分为应用胚胎脊髓和MK-801治疗组(A组),大鼠脊髓半切洞损伤应用胚胎脊髓移植组(B组),单纯脊髓半切洞损伤明胶海绵填充组(C组).手术后1、3、7、14 d对脊髓损伤区进行细胞凋亡的检测(TUNEL),以及Bcl-2蛋白免疫反应表达的测定(免疫组化法).采用计算机图像分析技术,进行定量分析.[结果]A、B、C三组中均发现凋亡细胞及BCL-2蛋白免疫反应阳性细胞,图像分析发现,细胞凋亡率为:C＞B＞A,Bcl-2免疫反应阳性细胞表达顺序为:A＞B＞C.[结论]大鼠脊髓损伤后应用NMDA受体拮抗剂MK-801和胚胎脊髓移植能抑制脊髓损伤后细胞凋亡.     

神经营养素-3对大鼠急性脊髓损伤后Bcl-2和Bax表达的影响

目的:观察神经营养素-3(NT-3)对大鼠急性脊髓损伤后B细胞淋巴瘤/白血病基因-2(Bcl-2)和B细胞淋巴瘤/白血病基因伴随蛋白x(Bax)表达的影响,探讨NT-3对脊髓损伤的作用及其可能的分子机制。方法:105只SD大鼠随机分为假手术组(A组)、损伤对照组(B组)和NT-3治疗组(C组),每组35只。B、C组用改良Allen′s法以30g·cm致伤大鼠T8脊髓制作损伤模型,C组经蛛网膜下腔导管于术后即刻、4h、8h、12h、24h、3d、7d注入NT-320μl(含NT-3200ng),B组在相同时间点给予等量生理盐水;A组打开椎板后蛛网膜下腔置管,不损伤脊髓,不给药。术后24h、3d、7d、14d对大鼠进行脊髓运动功能(BBB)评分。于术后4h、8h、12h、24h、3d、7d、14d处死动物(n=5),取T8节段脊髓,甲苯胺蓝(Nissl)染色观察脊髓前角运动神经元变化情况,用免疫组织化学方法检测Bcl-2和Bax在脊髓前角运动神经元中的表达变化情况。结果:各时间点C组和B组大鼠BBB评分均显著低于A组(P<0.01),但C组显著高于B组(P<0.05或0.01)。Nissl染色C组大鼠脊髓损伤区较B组出血少、残存神经元多,A组正常。各时间点C组和B组大鼠损伤脊髓前角运动神经元中Bax阳性细胞平均光密度(AOD)值均显著高于A组(P<0.01),但C组显著低于B组(P<0.05或0.01);各时间点C组和B组大鼠损伤脊髓前角运动神经元中Bcl-2阳性细胞AOD值均显著低于A组(P<0.01),但C组显著高于B组(P<0.05或0.01)。结论:NT-3可能通过抑制Bax表达,提高Bcl-2表达,抑制脊髓损伤后神经元凋亡,从而保护损伤的脊髓组织,这可能是NT-3对脊髓损伤具有保护作用的机制之一。     

Influence of an Aromatase Inhibitor (4-Acetoxy-4-Androstene-3,17-Dione) on Experimentally Induced Impairment of Spermatogenesis in Immature Rats/Einfluß eines Aromatasehemmers (4-Acetoxy-4-Androsten-3,17-dion) auf die experimentell induzierte Hemmung der Spermatogenese bei immaturen Ratten

Subcutaneous treatment of immature male rats with an estrogen precursor, 19-hydroxy-testosterone (19-OHT), at a daily dose of 1 mg/animal for 14 days leads to a significant decrease in the weight of testis, ventral prostate and seminal vesicle. The peripheral levels of LH are lowered. Testicular histology indicates that the effects of 19-OHT are very similar to the known of effect induced by estradiol-17 beta. 19OHT induces a marked impairment of spermato- and spermiogenesis. The maturation division is completely inhibited. The effect of 19-OHT on spermatogenesis is partially reversed by concomitant administration of an aromatase inhibitor (4-acetoxy-4-androstene-3.17-dione, (4-AA] at a dose of 1 mg/animal/day s.c. Meiotic activity is restored, and the weights of genital organs and the serum LH values increase. 4-AA alone has no appreciable effect on the parameters examined in this study. The present results suggest that specific inhibitors of estrogen biosynthesis might not only be useful to investigate the patho-physiological role of estrogens on spermatogenesis, but also be suitable to some extent for the treatment of estrogen-induced infertility in men suffering from idiopathic oligozoospermia.     

Amiloride and urinary trypsin inhibitor inhibit urothelial cancer invasion

OBJECTIVE: To determine the contribution of urokinase-type plasminogen activator (uPA) and plasmin in the invasion of highly invasive urothelial cancer cells. METHODS: We compared expression levels of mRNA and protease activity of uPA and plasmin formation in primary cultures of the noninvasive transitional cell carcinoma, UCT-1, and in the highly invasive type, UCT-2. By using in vitro cell invasion assay system, we evaluated the effects of amiloride and urinary trypsin inhibitor (UTI), which inhibit uPA and plasmin, respectively, on invasion by both cell lines. RESULTS: Expression levels of mRNA, protein, and activities of uPA were significantly higher (p<0.005) and resulted in more plasminogen activation in UCT-2 than in UCT-1. Amiloride and UTI significantly inhibited plasmin formation and the invasion of both cell lines (p<0.001). CONCLUSIONS: High expression levels of mRNA, activities of uPA and high plasmin formation significantly potentiated the invasiveness of urothelial cancer cells. Thus, inhibitors of uPA and plasmin, such as amiloride and UTI, respectively, could be useful therapeutic tools with which to treat urothelial cancer.     

内源性血管生成抑制因子的研究进展及在肝癌中的应用前景

目的 介绍内源性血管生成抑制因子（内皮抑素、血管抑素）在肝癌治疗中的作用和意义。方法 复习相关文献资料并作综述性报道。结果 内源性血管生成抑制因子通过抑制肿瘤血管的生长，有效地阻碍了肝癌的发展和转移，可为临床肝癌治疗提供新的途径。结论 应用内源性血管生成抑制因子的研究，对肝癌的防治将有积极的意义。     

A Kunitz Type of Proteinase Inhibitor Isolated from Boar Seminal Vesicle Fluid

Using immunoaffinity chromatography on a Sepharose 4B column with adsorbed antibodies to the basic inhibitor in bovine organs (Kunitz-type), a proteinase inhibitor was isolated from boar seminal vesicle fluid. The isolated protein inhibited acrosin, trypsin, plasmin and chymotrypsin, but not kallikrein. Its molecular weight determined by gel filtration on Sephadex G-50 was 9,500 (+/- 500) and by SDS electrophoresis in polyacrylamide gel 12,000 (+/- 500) daltons. The protein was demonstrated by immunoprecipitation only in boar seminal vesicle fluid and seminal plasma, and by indirect immunofluorescence on ejaculated spermatozoa and in the epithelium of boar seminal vesicles. This inhibitor is the first acrosin inhibitor specific for the genital organs, which evidently belongs to the group of Kunitz type inhibitors, to be described.     

Experience with belatacept rescue therapy in kidney transplant recipients

In kidney transplant recipients with chronic graft dysfunction, long‐term immunosuppression with calcineurin inhibitors (CNIs) or mTOR inhibitors (mTORi) can be challenging due to adverse effects, such as nephrotoxicity and proteinuria. Seventy‐nine kidney transplant recipients treated with CNI‐based or mTORi‐based maintenance immunosuppression who had CNI‐induced nephrotoxicity or severe adverse events were switched to belatacept. Mean time from transplantation to belatacept conversion was 69.0 months. Mean estimated glomerular filtration rate (eGFR) ± standard deviation at baseline was 26.1 ± 15.0 ml/min/1.73 m², increasing to 34.0 ± 15.2 ml/min/1.73 m² at 12 months postconversion (P < 0.0005). Renal function improvements were also seen in patients with low eGFR (<25 ml/min/1.73 m²) or high proteinuria (>500 mg/l) at conversion. The Kaplan–Meier estimates for patient and graft survival at 12 months were 95.0% and 85.6%, respectively. The discontinuation rate due to adverse events was 7.9%. One case of post‐transplant lymphoproliferative disorder occurred at 17 months postconversion. For comparison, a historical control group of 41 patients converted to mTORi‐based immunosuppression because of biopsy‐confirmed CNI‐induced toxicity was examined; eGFR increased from 27.6 ± 7.2 ml/min/1.73 m² at baseline to 31.1 ± 11.9 ml/min/1.73 m² at 12 months (P = 0.018). Belatacept‐based immunosuppression may be an alternative regimen for kidney transplant recipients with CNI‐ or mTORi‐induced toxicity.     

异丙酚对中枢多巴胺转运蛋白和5-羟色胺转运蛋白活性的影响

目的观察异丙酚对小鼠不同脑区中多巴胺转运蛋白和5羟色胺转运蛋白活性的影响,探讨异丙酚麻醉作用的机制.方法昆明小鼠27只随机分为3组,分别腹腔注射异丙酚100mg@kg-1、200mg@kg-1和同容积10%脂肪乳剂作对照,注射异丙酚10min后静脉注射对单胺类转运蛋白具有高度亲和力的放射性配基0.1ml2μCi125I-β-CIT,2h后动物断头处死,迅速分离小脑、下丘脑、纹状体和皮层,称湿重测脑区组织放射性,计算不同脑区组织与小脑放射性比值.结果异丙酚组纹状体/小脑-1值和皮层/小脑-1值与正常对照组相比明显下降,在异丙酚200mg@kg-1组降低更为明显(P＜0.01).结论异丙酚在中枢可降低125I-β-CIT与多巴胺转运蛋白和5羟色胺转运蛋白的结合,多巴胺转运蛋白和5-羟色胺转运蛋白可能参与了异丙酚麻醉的中枢作用机制.     

Timing of sirolimus conversion influences recovery of renal function in liver transplant recipients

Abstract: The long‐term use of calcineurin inhibitors (CNI) leads to renal dysfunction in many liver transplant (LT) recipients. The purpose of this analysis is to evaluate renal function in patients converted from CNI to sirolimus (SRL). From May 2002–November 2006, 137 LT were performed in 125 patients, 72 of which were converted to SRL. Evaluation of SRL conversion was stratified by early conversion (<90 d from LT) (EC) vs. late conversion (LC). Renal function was evaluated using the six‐point modification of diet in renal disease formula (estimated glomerular filtration rate [eGFR]). Forty‐two patients on SRL and 40 on CNI had at least three months of follow‐up and are included in the eGFR evaluation. At all time points after conversion, the EC group demonstrated a significantly higher mean eGFR than those in the LC group. A significant improvement in eGFR was seen within the EC group when comparing eGFR at time of conversion to eGFR at three, six, nine, and 12 months after conversion and last follow‐up. The only improvement in the LC group was from conversion to the three‐month time point. We conclude that EC to SRL results in a profound improvement in eGFR that begins at three months and is sustained beyond one yr.     

Combining immunotherapies for the treatment of prostate cancer

Sipuleucel-T, a therapeutic dendritic-cell vaccine, was Food and Drug Administration–approved for prostate cancer in 2010. No new immunotherapies for prostate cancer have been approved since. However, novel agents and combination approaches offer great promise for improving outcomes for prostate cancer patients. Here we review the latest developments in immunotherapy for prostate cancer. Sipuleucel-T has demonstrated a survival advantage of 4.1 months in metastatic castration-resistant prostate cancer. PSA-TRICOM (PROSTVAC), a prostate-specific antigen–targeted vaccine platform, showed evidence of clinical and immunologic efficacy in early-phase clinical trials, and results from a phase III trial in advanced disease are pending. While immune checkpoint inhibitors appear to have modest activity as monotherapy, preclinical and clinical data suggest that they may synergize with vaccines, poly [ADP-ribose] polymerase inhibitors, and other agents. Several clinical studies that combine these therapies are underway.Combining prostate cancer vaccines with immune checkpoint inhibitors has great potential for improving clinical outcomes in prostate cancer. Such combination approaches may create and then recruit tumor-specific T cells to tumor while also increasing their effector function. Other emerging agents may also enhance immune-mediated tumor destruction.