胸腺内注射脾细胞诱导神经移植的免疫耐受反应

背景：心脏等移植时常采用胸腺内注射脾细胞抑制或减弱移植排斥反应，而在神经移植中很少有报道。 目的：应用大鼠胸腺内注射脾细胞的方法，诱导大鼠同种异体坐骨神经产生特异性免疫耐受。 设计、时间及地点：随机对照动物实验，于2007-06/2008-06在哈尔滨医科大学动物实验中心完成。 材料：选用清洁级雄性SD大鼠30只为受体，随机分为自体神经移植组，异体神经移植组，异基因抗原注射组，每组10只。雄性Wistar大鼠20只为供体。 方法：异体神经移植组大鼠在显微镜下，从犁状肌下孔0.5 cm处整齐剪下长约1 cm的坐骨神经，将供体神经桥接于神经缺损处。自体神经移植组神经缺损处进行自体神经移植。异基因抗原注射组在异体神经移植后注入供体异基因抗原。 主要观察指标：移植后2周进行运动神经传导速度、病理学、混合淋巴细胞培养和血清白细胞介素检查。 结果：运动神经传导速度异基因抗原注射组与自体神经移植组无显著差别(P > 0.05)，但明显优于异体神经移植组（P < 0.05）。病理学及透射电镜观察，与运动神经传导速度检测结果一致。混合淋巴细胞培养和白细胞介素水平异基因抗原注射组明显优于异体神经移植组（P < 0.05）。 结论：胸腺内注射异基因抗原可以诱导大鼠对异体坐骨神经移植的免疫耐受。     

同种异体肢体移植特异性免疫耐受诱导方案

背景：通过诱导受体产生免疫耐受能够解决移植领域存在的许多问题,如长期应用免疫抑制剂带来的毒副作用,慢性排斥反应等。 目的：观察联合应用亚致死量放射线照射、氟达拉滨及骨髓腔内骨髓移植对诱导大鼠同种异体肢体移植产生特异性免疫耐受的影响。 设计、时间及地点：随机同期对照动物实验,于2004-01/2008-12在哈尔滨医科大学完成。 材料：40只雄性Wistar大鼠随机分成4组：单纯移植组、氟达拉滨组、骨髓移植组、氟达拉滨＋骨髓移植组,每组各10只。40只SD大鼠为骨髓移植供体。供体DA大鼠用于皮肤移植实验。 方法：制作同种异体右后肢移植模型,单纯移植组仅进行肢体移植;氟达拉滨组在肢体移植前1 d,受体给予亚致死量60Co照射及腹腔内注射氟达拉滨50 mg/kg;骨髓移植组肢体移植当天受体接受供体骨髓腔内骨髓移植1×1012 L-1,10 μL;氟达拉滨+骨髓移植组受体联合应用亚致死量照射、注射氟达拉滨及骨髓腔内骨髓移植。 主要观察指标：观察移植物排斥反应征象及存活情况,并对氟达拉滨+骨髓移植组产生免疫耐受大鼠进行SD大鼠及DA大鼠皮肤移植及脾细胞中细胞因子mRNA检测。 结果：与其他实验组比较,氟达拉滨＋骨髓移植组移植物发生排斥反应的时间及存活时间均显著延长(P < 0.01)。氟达拉滨+骨髓移植组大鼠仅对第3方DA大鼠的皮肤呈现强烈的免疫反应,移植皮肤初为水疱、渗出,逐渐溃疡、糜烂、焦痂、变黑坏死。 与单纯移植组比较,氟达拉滨+骨髓移植组大鼠Th1型细胞因子的表达明显降低,而Th2型细胞因子的表达明显增高。 结论：氟达拉滨与骨髓移植联合方案可以诱导大鼠同种异体肢体移植产生特异性免疫耐受。     

新鲜自体与异体骨软骨移植修复兔关节软骨缺损的比较

背景：骨软骨移植是修复关节软骨损伤的主要方法之一,但新鲜自体骨软骨移植与新鲜异体骨软骨移植相比效果尚无定论。 目的：探讨新鲜自体骨软骨移植与异体骨软骨移植修复兔关节软骨缺损的方法及疗效。 设计、时间及单位：动物实验观察,于2007-01/2008-05在解放军第四军医大学唐都医院骨科完成。 材料：新西兰大白兔9只,随机分为3组,3只/组,分别实施自体骨软骨移植、异体骨软骨移植、单纯软骨缺损3种手术。 方法：自体移植组：在股骨内侧髁负重关节面取直径3 mm骨软骨缺损,于非负重关节面取4块直径1 mm骨软骨柱移植;异体移植组：在股骨内侧髁负重关节面直径3 mm骨软骨缺损处植入相同直径的异体骨软骨柱;对照组：于股骨内侧髁负重关节面取直径3 mm全层软骨缺损,不做处理。 主要观察指标：术后12周取材,进行大体观察,组织学检查和半定量的改良Wakitani score评分。 结果：大体观察异体移植组修复面较自体移植组稍平整,光镜可见自体和异体移植软骨均已覆盖缺损,与正常软骨高度相当。绝大部分修复面为透明软骨,增殖活跃。软骨下骨、松质骨小梁均与骨床完全骨性愈合。自体移植组软骨浅表区细胞数目及排布更接近正常透明软骨,过渡区及辐射区细胞密度、厚度也明显优于异体移植组,基质染色丰富。 结论：自体与异体骨软骨移植的方法均可完成关节软骨缺损的透明软骨修复,自体骨软骨移植更优。     

异体颈动脉体球细胞脑内移植对帕金森病大鼠行为及纹状体多巴胺含量的改善

帕金森病（Parkinson disease,PD)是一种好发于中老年人的中枢神经系统退行性疾病,细胞移植治疗是一项重要的手段,近年来,有研究表明颈动脉体球细胞在低氧状态下生长良好,且具有旺盛的分泌多巴胺（DA）的特性,为此,我们利用颈动脉体球细胞作为移植组织来源,系统观察移植治疗偏侧PD大鼠行为及DA的改善情况。     

同种异基因大鼠间充质干细胞移植对体内CD4+CD25+T细胞的影响**☆

背景：间充质干细胞的免疫抑制作用近年来逐渐得到证实并开始应用于临床,但相关研究成果主要来源于体外细胞实验。 目的：观察同种异基因大鼠骨髓间充质干细胞从静脉输入后对体内CD4+CD25+调节性T细胞的影响。 设计、时间及地点：以动物为对象的对照观察实验,于2008-03/09在南方医科大学珠江医院移植免疫研究所完成。 材料：Wistar大鼠6只用于制备骨髓间充质干细胞,SD大鼠20只作为输注骨髓间充质干细胞的受体鼠。 方法：从Wistar大鼠骨髓分离培养间充质干细胞,取第3~5代细胞进行实验。①体外淋巴细胞增殖实验：首先将间充质干细胞悬液从尾静脉注入SD大鼠体内10 d后脱臼处死,取大鼠脾脏制备脾淋巴细胞。实验分5组：分别为脾淋巴细胞/骨髓间充质干细胞为1∶10,1∶50,1∶100+ConA 5 μg组,脾淋巴细胞+ConA 5μg组（增殖组）,单纯淋巴细胞组(空白组),检测共培养体系中CD4+CD25+ / CD4+ T淋巴细胞比率。②体内注射骨髓间充质干细胞实验：将5×109 L-1,5×108 L-1,5×107 L-1间充质干细胞及PBS静脉输入SD大鼠体内,10 d后取受体鼠胸腺、脾脏、外周血检测CD4+CD25+ / CD4+ T淋巴细胞比率。 主要观察指标：①淋巴细胞增殖体系中CD4+CD25+/CD4+比率的变化。②SD大鼠胸腺、脾脏、外周血CD4+CD25+/CD4+比率的变化。 结果: ①体外淋巴细胞共培养体系中,1∶10组T细胞亚群CD4+ CD25+ / CD4+细胞百分率较增殖组显著升高（P < 0.01）。②体内输注间充质干细胞达5×109 L-1的受体鼠外周血和脾脏CD4+CD25+ / CD4+ T细胞比率上升,与输注PBS组相比有显著差异（P < 0.05）,而在胸腺中各组比率无明显差异。 结论：高浓度同种异基因大鼠间充质干细胞不仅可以在体外实验中增加CD4+CD25+ / CD4+ T细胞比率,静脉输入后仍具有相同作用。     

患儿鞘内注射化疗药物的护理体会

目的 总结23例患儿184次鞘内注射化疗药物的护理要点。方法 通过强调术前准备,熟悉鞘内注射治疗过程,注意预防不良反应。结果 不良反应发生率低,未出现严重不良反应。结论 采取合适的护理方法可有效提高鞘内注射成功率,减少不良反应的发生率。     

输注同基因骨髓间充质干细胞联合供体脾组织移植诱导肝移植大鼠免疫耐受的研究

摘要：目的 观察输注与受体同基因骨髓间充质干细胞联合供体脾组织移植对诱导大鼠肝移植术后免疫耐受作用,探讨其可能的机制。方法 实验鼠随机分为4组,每组24只。A组行DA-Lewis大鼠原位肝移植,B组行DA-Lewis大鼠原位肝移植术后予口服环孢素,C组行DA-Lewis大鼠原位肝移植同期输注Lewis大鼠骨髓间充质干细胞。D组在C组的基础上术中同期行DA大鼠脾组织移植。观察各组生存期,肝功能情况,血清细胞因子水平,嵌合体的形成情况及肝脏的病理变化。结果 与其他各组相比,D组大鼠存活时间明显延长,术后D组血清ALT、AST、TBIL较A组显著降低,与B、C组相近;B、C、D组IL-2、IFN－γ均升高,但低于A组（P<0.05）;、而IL-4和IL-10高于A组（P<0.05）,B、C组移植肝仅呈急性轻度排斥反应,A组呈急性重度排斥反应,D组未见明显排斥反应。30d后D组受体脾脏中供体阳性细胞明显高于各组（P<0.05）。结论 大鼠肝脏脾组织移植后输注与受体同基因骨髓间充质干细胞后能减轻移植肝的排斥作用,甚至诱导免疫耐受。     

中枢神经系统白血病鞘内注射化疗的护理

随着医学的进展,白血病完全缓解率逐年升高,生存期延长。但由于多种化疗药物不易透过血脑屏障,致使中枢神经系统成为白血病的庇护所。中枢神经系统白血病(CNSL)可发生在白血病的任何阶段,但少见于发病初期。     

静脉滴注联合鞘内/脑室内注射多粘菌素治疗颅内多重耐药菌感染有效性和安全性的Meta分析

目的 评价静脉滴注(IV)联合鞘内/脑室内注射(ITH/IVT)多粘菌素治疗颅内多重耐药菌感染的有效性及安全性。方法 计算机检索万方数据库、维普中文科技期刊数据库、中国生物医学文献数据库、Pubmed、Embase、ScienceDirect及Cochrane Library,筛选2023年1月前有关IV+ITH/IVT多粘菌素治疗颅内多重耐药菌感染的队列研究。采用RevMan 5.4软件进行Meta分析。结果 最终纳入9项回顾性临床研究。IV+ITH/IVT组病死率显著低于IV组(OR=0.19,95%CI:0.11～0.35,P<0.01)。IV+ITH/IVT组与IV组ICU住院天数(OR=-2.32,95%CI:-23.59～18.89,P=0.83)及不良反应率(OR=0.93,95%CI:0.26～3.38,P=0.91)差异无统计学意义。结论 IV+ITH/IVT多粘菌素治疗颅内多重耐药菌感染可以显著降低患者病死率,且不显著增加不良反应。     

注射鼠神经生长因子对痉挛型脑瘫运动功能的改善作用

目的观察不同途径注射鼠神经生长因子(恩经复)改善痉挛型脑瘫运动功能的效果。方法在河南省内3家医疗单位采用统一标准,对照组296例患儿给予综合康复治疗,3周一疗程,每疗程结束后休息2周,共治疗3个疗程。穴位注射组281例以及肌内注射组287例患儿在综合康复治疗的基础上分别于每周二、四、六给予穴位及肌内注射鼠神经生长因子,每疗程10次,每疗程结束后休息2周,开始下一疗程治疗,共计3个疗程。每疗程均进行疗效评估。结果穴位注射组与肌内注射组在降低肌张力(Ashworth评分、sEMG数值)、提高运动功能(盖泽尔评分、GMFM评分)方面均优于对照组(P0.05),穴位注射组优于肌内注射组(P0.05)。结论对于3岁以下痉挛型脑瘫患儿,穴位注射鼠神经生长因子安全有效。     

Nerve allograft regeneration and immunological tolerance of rats with sciatic nerve transplantation after intragastric pretreatment with ultraviolet B-irradiated donor splenocytes★

BACKGROUND: Nerve allograft rejection is an unavoidable problem for nerve allografts. Traumatic peripheral nerve injuries are commonly reconstructed using autogenous nerve grafts. However, this form of reconstruction is limited by insufficient autologous nerves for large gap repairs and by morbidity at the nerve donor site. OBJECTIVE: To examine sciatic nerve regeneration and immune tolerance reaction after intragastric administration of ultraviolet B-irradiated (UVB) donor splenocytes. DESIGN, TIME AND SETTING: A complete randomized grouping design and controlled experiment. The experiments were conducted in the Department of Orthopedics, the First Affiliated Hospital to Shanxi Medical University, China, between March and October 2007. MATERIALS: Fourteen adult male SD rats and fourteen male Wistar rats, weighing 250–300 g, were randomly matched as donors and acceptors. A further seven male SD rats (weight 250–300 g, age 12–16 weeks) were used for nerve isografts. Immune preparations and the Epics XL flow cytometer were purchased from B-D Company, USA. A computer-assisted electromyograph machine was provided by Keypoint P, Dantel Company, Denmark. METHODS: Splenocytes from Wistar rats were isolated, purified, and cultured, and then irradiated with ultraviolet B. In the first control group (Group 1), the SD rats received a syngeneic SD nerve isograft. In the second control group (Group 2), the SD rats received a nerve allograft from Wistar rats without pretreatment. In the oral-tolerance treated group (Group 3), the SD recipient rats were inoculated with 2.5×107 Lewis UVB-irradiated donor splenocyte cells by intestinal tract administration at seven days before transplantation. MAIN OUTCOME MEASURES: (1) The recent end and the middle and distal end of the transplanted nerve were cut at 8 and 12 weeks after operation. Recovery of nerve regeneration was measured with HE staining using the total number, density, and diameter of the nerve fibers. (2) The level of CD25+T lymphocytes in peripheral blood was detected with the Epics XL flow cytometer at one week after operation. (3) The bilateral sciatic nerves were exposed from the sciatic notch up to 0.5 mm beyond the distal graft site at eight weeks post-engraftment. Bipolar platinum stimulating electrodes were placed under the sciatic nerve proximally and the mean conduction velocity was recorded with recording electrodes on the posterior tibial nerve at 0.3 cm distal to the nerve graft. RESULTS: Eight weeks after operation, total axon number and fiber density in Group 3 were higher than that in Group 1 (P < 0.05), neural regeneration in Group 3 was lower than that in Group 1 (P < 0.05) , The level of CD25+T lymphocytes in peripheral blood of Group 3 was significantly lower than that of Group 2 (P < 0.05). There was no significant difference between Group 3 and Group 1 (P > 0.05). At eight weeks post-engraftment the mean conduction velocity of Group 3 approximated that of Group 1. The untreated allografts in Group 2 demonstrated no measurable recovery of conduction velocity. CONCLUSION: Pretreatment with a single intragastric dose of UVB-modified donor antigen specifically induced tolerance to peripheral nerve allografts in rats. Key Words: sciatic nerve; transplantation, homologous; immune tolerance     

Repeated low doses of morphine induce a rapid tolerance in arthritic rats but a potentiation of opiate analgesia in normal animals

The effect of repeated low doses of morphine (0.3–3 mg/kg s.c., twice daily for 4 days) on subsequent sensitivity to the antinociceptive effect of morphine was tested in arthritic and normal rats. Chronic morphine induces tolerance in arthritic rats. This tolerance develops rapidly since it is clearly present after one day of treatment. By contrast, in normal animals similarily treated, a potentiation of morphine analgesia was observed.     

Effect of intrathymic injection of allogene antigen on immune response to sciatic nerve transplantation in allogenic mice

BACKGROUND: The latest researches demonstrate that intrathymic injection of MHC antigen which reaches a certain dosage (2 mg, i.e., 4 × 108 cell extraction) can induce immunologic tolerance under non-antilymphocyte serum condition. OBJECTIVE: To investigate the effect of intrathymic injection of allogene antigen on survival and function of sciatic nerve in allogenic mice. DESIGN: Randomized controlled animal study. SETTING: The 4th Affiliated Hosptial of Harbin Medical University. MATERIALS: A total of 32 male donor C57BL/6(H-2b) mice of 4–8 weeks old and weighing 18–22 g and 44 female receptor Balb/c(H-2d) mice of 4–8 weeks old and weighing 18–22 g were selected from Heilongjing Veterinary Institution. The animal experiment had got confirmed consent from local ethic committee. METHODS: The experiment was carried out in the Laboratory (Provincial Key Laboratory) of the Fourth Hospital, Harbin Medical University from June 2006 to May 2007. C57BL/6(H-2b) mice were anesthetized to extract MHC (H-2b) antigen from splenic cells and sciatic nerves. Allogenous nerve transplantation group: Mice were given intrathymic injection of 100 μL saline; two weeks later, frozen sciatic nerves of donor mice were transplanted. Immunosuppressive agent group: Mice were given intrathymic injection of 100 μL saline; two weeks later, fresh sciatic nerves of donor mice were transplanted. At three days before transplantation, 10 mg/kg per day cyclosporin A was intraperitoneally injected once a day till mice were sacrificed. MHC (H-2b) antigen injection group: Mice were given intrathymic injection of MHC (H-2b) antigen from C57BL/6(H-2b) donor mice; two weeks later, fresh sciatic nerves of donor mice were transplanted. Autogenous nerve transplantation group: Mice were given intrathymic injection of 100 μL saline; two weeks later, fresh sciatic nerves were transplanted. MAIN OUTCOME MEASURES: ① Three weeks later, transplanted part of exposured sciatic nerve was used to measure the motor nerve conduction velocity. ② Transplanted nerve was stained with histochemical staining and observed light microscope and electron microscope. ③ Mice received mixed lymphocyte culture and delayed-typed hypersensitiveness to observe absorbency and measure depth of soles. RESULTS: All 76 mice were involved in the final analysis. ① Motor nerve conduction velocity: The nerve recovery in MHC (H-2b) antigen injection group was higher than that in allogenous nerve transplantation group, equal to immunosuppressive agent group and lower than autogenous nerve transplantation group. There were significant differences among them (P < 0.05). ② Histological changes of transplanted nerve: Light and electron microscopes demonstrated that there were a lot of regenerative nerve fibers in autogenous nerve transplantation group, immunosuppressive agent group and MHC (H-2b) antigen injection group, and all nerve fibers passed grafts. ③ Immunological examination: There was no significant difference in mixed lymphocyte culture among allogenous nerve transplantation group, autogenous nerve transplantation group and MHC (H-2b) antigen injection group (P < 0.05). Depth of soles in other groups was deeper than that in the MHC (H-2b) antigen injection group, and there was significant difference (P < 0.05); that was to say, delayed-typed hypersensitiveness was remarkable. CONCLUSION: The intrathymic injection of allogene MHC antigen may induce specific immune tolerance to allogenous sciatic nerve transplantation and promote nerve survival and functional recovery.     

缺血预处理上调TROY表达诱导大鼠局灶性脑缺血耐受

目的研究缺血预处理（IPC）对局灶性脑缺血的保护作用以及对TNFRSF expressed on the mouse embryo（TROY）表达的影响。方法33只雄性成年SD大鼠，随机分为预处理6h组（IVC-6组，11只）、预处理72h组（IPC-72组，11只）和缺血组（CI组，11只）。利用线栓法建立大脑中动脉栓塞（MCAO）致局灶性脑缺血模型。MCAO 10min作为IPC,IPC-6和IPC-72组分别在IPC后6h和72h制作短暂性局灶性脑缺血模型。再灌注24h后，对所有动物行神经功能缺损评分（NDS），并处死大鼠取脑，应用2％TTC染色测定梗死容积、TUNEL染色研究细胞凋亡情况和免疫组化观察TROY表达变化。结果与CI组比较，IPC-72组显著改善局灶性脑缺血24h后大鼠神经功能损害[两组评分分别为2（1．5-3），1（0—2）]，减少脑梗死容积[（299．33±70．98）mm^3，（69．25±47．66）mm^3]，抑制细胞凋亡和增强TROY的表达（阳性细胞数分别为42±11，87±17）（P〈0．01）。结论IPC对其后局灶性脑缺血有明显的保护作用．能诱导脑缺血耐受，可能与TROY表达上调相关。     

Different chronic ethanol exposure regimens in adolescent and adult male rats: effects on tolerance to ethanol-induced motor impairment

Findings are mixed regarding the expression of tolerance after repeated ethanol exposure, perhaps in part due to dose/frequency variations in exposure regimens. The present study compared age-related differences in tolerance development following 10 days of 1 g/kg twice daily, 2 g/kg once daily, or intermittent 4 g/kg ethanol exposure regimens. To measure expression of chronic tolerance and acute tolerance, ethanol-induced motor impairment was assessed on day 12, with functionally equivalent ethanol doses administered across age (2 g/kg - adolescents; 1.5 g/kg - adults). Subsequent challenge doses resulted in lower brain ethanol concentrations in both age groups as a function of the chronic ethanol regimens. Expected age-related differences emerged in acute tolerance expression in non-manipulated animals, with adolescents, but not adults showing acute tolerance. Regimens sufficient to induce alterations in ethanol metabolism did not result in chronic functional tolerance at either age, although chronic injections were sufficient to induce acute tolerance in adults.     

未成熟髓源树突状细胞负载P2 58-73肽段诱导免疫耐受对实验性自身免疫性神经炎的预防作用

目的观察未成熟髓源树突状细胞(iMDC)负载P258-73肽段诱导免疫耐受对实验性自身免疫性神经炎(EAN)的预防作用,以及对干扰素-γ(IFN-γ)、白介素-33(IL-33)mRNA表达的影响。方法(1)P258-73aa与iMDC共培养。(2)21只Lewis大鼠随机分为EAN组(A组)、iMDC组(B组)和P258-73aa-iMDC组(C组),并分别于皮下注射磷酸盐缓冲液(PBS)、iMDC及P258-73aa-iMDC;7d后,各组均给予P253-78aa和完全弗氏佐剂(CFA)进行免疫,诱发EAN。观察各组发病情况并作临床评分至免疫后16d(发病高峰期)。(3)采用3H-TdR掺入法检测淋巴细胞增殖反应;RT-PCR检测大鼠坐骨神经、脾脏和淋巴结中IL-33、IFN-γmRNA的表达。结果(1)发病高峰期,A组、B组、C组的临床评分为(7.4±1.9)分、(5.2±1.6)分和(3.4±0.9)分,各组间比较差异有统计学意义(均P<0.01)。(2)A组、B组、C组抗原特异性淋巴细胞增殖反应依次明显降低(均P<0.01)。(3)A组、B组、C组坐骨神经、脾脏和淋巴结中IFN-γmRNA表达水平依...     

自制旋转持续静脉输液装置在大鼠同种异体小肠移植排斥模型构建中的应用*☆

背景：构建大鼠小肠移植模型是研究小肠移植排斥反应的重要基础,实验中对动物无法进行长期持续的静脉输液成为延误实验顺利开展的最大障碍。 目的：将自制旋转持续静脉输液装置与大鼠异位小肠移植模型相结合,验证输液装置的可靠性和动物模型的稳定性。 设计、时间及地点：随机区组,对照观察实验,于2008-03/06在解放军第四军医大学西京医院消化病研究所全军医学重点实验室完成。 材料：选用健康雄性近交系F344/NCrl BR大鼠48只为供体,LEW/Crl 大鼠48只为受体,建立同种异体节段性异位小肠移植模型。 方法：将移植后的大鼠以随机化完全区组设计分为3组(n=16)：对照组、输液组和他克莫司治疗组。对照组大鼠仅行小肠移植,移植前后不做持续静脉输液。输液组大鼠给予持续输注肠外营养液。他克莫司治疗组持续输注肠外营养液+静脉注射他克莫司0.5 mg/(kg•d)。 主要观察指标：移植后观察移植大鼠的一般状况和存活时间,并于移植后3,5,7 d 分别取各组大鼠移植肠标本(n=3)进行组织病理学检查,每组剩余的大鼠作存活时间观察,观察期限为5周。 结果：对照组大鼠平均存活时间为(5.4±1.7) d,手术成功率56.3%。采用旋转输液装置的输液组和他克莫司治疗组大鼠手术成功率为90.6%,与对照组比较,差异有显著性意义(P < 0.01)。输液组大鼠平均存活(7.2±2.8) d,最终全部死于急性排斥反应。他克莫司治疗组剩余大鼠的输液时间> 30 d,存活时间超过5周,与对照组和输液组比较,差异有统计学意义(P < 0.05)。对照组和输液组大鼠术后3,5,7 d移植肠组织病理学检查分别符合轻、中、重度排斥反应。他克莫司治疗组大鼠术后3,5,7 d未见明显排斥反应征象。 结论：旋转持续静脉输液装置制作简单,固定安全可靠,能成功地为研究大鼠连续输液30 d以上,与异位小肠移植模型联合应用,能建立稳定可靠的排斥反应模型。     

Possible role of the superoxide anion in the development of neuronal tolerance following ischaemic preconditioning in rats

There is a large body of evidence that reactive oxygen species play a major role in the pathogenesis of ischaemic brain damage. On the other hand, it has recently been suggested that superoxide anions participate in the development of neuronal tolerance against lethal ischaemia following ischaemic preconditioning (PC). The present study aimed to examine whether or not the intravenous administration of human recombinant Cu/Zn superoxide dismutase (hr SOD) prior to PC would affect the subsequent development of neuronal tolerance. Animals were randomly assigned to the following three groups: group 1, sham PC treated with vehicle; group 2, PC treated with hr SOD and group 3, PC treated with vehicle. For PC, 10 min occlusion of the middle cerebral artery (MCA) by a modified intraluminal suture method was followed by 60 min recirculation and this procedure was successively repeated three times. The procedures were similar for sham PC except that the MCA was kept unoccluded. Just prior to PC or sham PC, a bolus of hr SOD (6 x 103 IU/2 ml/kg) was administered intravenously. Seventy-two hours thereafter, rats were subjected to lethal ischaemia, i.e. MCA occlusion for 100 min followed by recirculation for 48 h. The infarct area and volume were assessed with the 2,3,5-triphenyltetrazolium stain. A significant difference in the infarct volume was revealed between the sham PC/vehicle and the PC/vehicle groups (total and cortex P < 0.01; striatum P < 0.05), showing that PC induced a marked neuronal tolerance against lethal ischaemia. The infarct volume in the PC/SOD group was close to that in the sham PC/vehicle group, being significantly greater than that in the PC/vehicle group (total and cortex P < 0.01) and showing that the administration of hr SOD suppressed the development of neuronal tolerance induced by PC. In a parallel experiment, expression of 72-kDa heat-shock protein (hsp 72) at 72 h after PC was considerably reduced in rats treated with hr SOD compared with those treated with vehicle. These results suggest that superoxide anions intraluminally generated within cerebral microvessels participate in the development of neuronal tolerance as well as the induction of hsp 72 following PC.     

大鼠脑缺血耐受与脑自体神经干细胞的增殖

背景：脑缺血耐受与脑自体神经干细胞均具有脑保护作用,但前者能否促使脑自体神经干细胞增殖,学者们报道不一致。 目的：明确缺血预处理与大鼠脑梗死后7 d海马区自体神经干细胞增殖的关系,以及其对大鼠脑梗死后神经行为学评分的影响。 方法：采用二次线栓法建立局灶-局灶性SD大鼠脑缺血耐受模型,40只SD雄性大鼠随机分为：假手术组,缺血组,假手术+缺血组,预缺血+缺血组,每组10只。脑梗死后3,7 d采用Zea-Longa评分方法进行神经行为学评分,运用荧光免疫组织化学技术检测大鼠脑缺血侧海马区BrdU标记阳性细胞数量。 结果与结论：脑梗死后3,7 d的Zea-Longa神经行为学评分,预缺血+缺血组低于缺血组、假手术+缺血组(P < 0.01),而缺血组和假手术+缺血组间差异无显著性意义(P > 0.05)。脑梗死后7 d缺血侧海马区BrdU标记阳性细胞数,缺血组、假手术+缺血组、预缺血+缺血组高于假手术组(P < 0.01);预缺血+缺血组高于缺血组、假手术+缺血组(P < 0.01);而缺血组和假手术+缺血组间差异无显著性意义(P > 0.05)。结果表明缺血预处理可促进大鼠脑梗死后海马区齿状回颗粒下层成体神经干细胞的增殖,并能改善其神经功能缺损症状。     

Involvement of protein kinase C in morphine tolerance at spinal levels of rats

The present study was performed to investigate the possible role of protein kinase C (PKC) in morphine tolerance at spinal levels of rats. Intrathecal injection of 10 μg of morphine induced increases in the hindpaw withdrawal latency (HWL) to noxious thermal and mechanical stimulation in rats. After intrathecal injections of 10 μg of morphine (twice a day) lasted for 5 days, the antinociceptive effects induced by intrathecal injections of morphine decreased significantly in rats. Interestingly, we found that there were significant increases in the content of PKC in the dorsal horn of the spinal cord and the dorsal root ganglion, but not in the ventral horn of the spinal cord, in rats with morphine tolerance determined by Western blot, suggesting that PKC is involved in morphine tolerance at spinal levels of rats. Furthermore, our results demonstrated that chronic intrathecal injection of the PKC inhibitor significantly inhibited the development of morphine tolerance. Moreover, we found that the maintenance of morphine tolerance was blocked by intrathecal administration of a PKC inhibitor in rats, and the inhibitory effects of the PKC inhibitor on morphine tolerance lasted for more than two days. Taken together, the present study clearly showed that PKC is involved in morphine tolerance at the spinal level of rats and that intrathecal administration of a PKC inhibitor can block the development and maintenance of morphine tolerance.