同种原位心脏移植的免疫抑制治疗

对２例同种原位以及移植受者采用环孢素Ａ（ＣｓＡ）、硫唑嘌呤（Ａｚａ）及皮质类固醇三联疗法进行免疫抑制治疗，治疗分为围手术期及长期维持两个阶段。围手术期的免疫抑制剂用量较大，１周后逐渐减量，ＣｓＡ１２个月后达维持量（２－３ｍｇ，ｋｇ＾－１／ｄ），Ａｚａ术后４０周达维持量（１．２－１．５ｍｇ．ｋｇ＾－１／ｄ），皮质粝固醇１年后停用。例１、例２分别发生３次，１急性排斥反应，经冲击治疗逆转。２例患者用药后     

小剂量氯胺酮吗啡及氯胺酮-吗啡配伍用于硬膜外腔术后镇痛作用的比较小剂量氯胺酮吗啡及氯胺酮-吗啡配伍用于硬膜外腔术后镇痛作用的比较

目的：探讨氯胺酮、吗啡硬膜外腔术后镇痛效应和伍用后是否可提高镇痛效果并减少副作用。方法：５０例硬膜外腔麻醉下行骨科手术的患者，随机分为５组，每组１０例。Ａ组：吗啡０．０１ｍｇ／ｋｇ；Ｂ组：氯胺酮０．４ｍｇ／ｋｇ；Ｃ组氯胺酮０．６ｍｇ／ｋｇ；Ｄ组：Ａ＋Ｂ；Ｅ组：Ａ＋Ｃ。于术后４、８、１２、２４、４８、７２ｈ记录疼痛评分（ＶＡＰＳ）及副作用的发生情况。结果：Ａ组ＶＡＰＳ评分平均为２．９５，有效镇痛７例，平均持续时间为５２．０ｈ；Ｂ组镇痛效果差，ＶＡＰＳ评分平均为７．２６，有效镇痛３例，与Ａ组比较有统计学显著差异（Ｐ＜０．０１）；Ｃ组ＶＡＰＳ评分平均为３．６０，与Ａ组比较无统计学差异，有效镇痛７例，平均持续时间为４４．４ｈ；Ｄ组ＶＡＰＳ平均评分为２．７３，与Ａ组比较无统计学差异，平均持续时间为５０．８ｈ；Ｅ组平均ＶＡＰＳ评分为１．５８，与Ａ组比较有统计学显著差异（Ｐ＜０．０１），持续时间为５８．１ｈ。结论：１．氯胺酮０．４ｍｇ／ｋｇ硬膜外腔术后镇痛效果差，剂量增至０．６ｍｇ／ｋｇ镇痛效果与吗啡０．０１ｍｇ／ｋｇ相近，恶心、呕吐发生较少，无精神方面的副作用；２．氯胺酮与吗啡配伍，随着氯胺酮剂量增加到０．６ｍｇ／ｋｇ     

雷公藤多甙在大鼠肾移植模型中的实验研究

用显微外科技术建立大鼠肾移植模型（Ｗ→ＳＤ）。将大鼠分为四组：对照组仅给生理盐水，雷公藤多甙（Ｔ_Ⅱ）组给Ｔ_Ⅱ３０ｍｇ·ｋｇ~（１）·ｄ~（－１），ＣｓＡ组给ＣｓＡ１５ｍｇ·ｋｇ~（－１）·ｄ~（－１），ＣｓＡ＋Ｔ_Ⅱ组同时给ＣｓＡ１５ｍｇ·ｋｇ~（－１）·ｄ~（－１）＋Ｔ_Ⅱ３０ｍｇ·ｋｇ~（－１）·ｄ~（－１）。对照组大鼠肾移植后平均存活时间为７．３±２．２天，Ｔ_Ⅱ组平均存活时间为１８．１±３．７天，两组存活时间有显著性差异（Ｐ＜０．０５）。Ｔ_Ⅱ＋ＣｓＡ组平均存活时间为３４．６±５．２天，ＣｓＡ组为２６．１±４．６天。单纯用药的两组分别与联合用药组比较均有显著性差异（Ｐ＜０．０５）。Ｔ_Ⅱ３０ｍｇ·ｋｇ~（－１）·ｄ~（－１）对大鼠肝、肾、心脏及白细胞总数无影响，但显著抑制大鼠脾淋巴细胞的转化。Ｔ_Ⅱ也显著抑制大鼠外周血白细胞介素－２受体水平，但抑制能力不及ＣｓＡ。     

诱发性一氧化氮合酶抑制药对感染性休克鼠肝,肺组织学改变？…

目的 研究诱导性一氧化氮合酶（ｉＮＯＳ）抑制药氨基胍（ＡＧ）和非选择性ＮＯＳ抑制药Ｌ－Ｎ硝基精氨甲酯（Ｌ－ＮＡＭＥ）对感染性休克鼠肝肺组织学改变的影响。方法 小鼠腹腔内注射（ｉ．ｐ．）内毒素（ＬＰＳ,２０ｍｇ·ｋｇ＾－１）后４小时随机分为ＬＰＳ组（ｎ＝６０）,ＬＰＳ＋Ｌ－ＮＡＭＥ组（３０ｍｇ·ｋｇ＾－１ｉ．ｐ．,ｎ＝６０）。和ＬＰＳ＋ＡＧ组（２０ｍｇ·ｋｇ＾－１ｉ．ｐ．ｎ＝６０）。５小时后取受试鼠     

不同剂量异丙酚对脑血流动力学的影响

目的：采用经颅多普勒（ＴＣＤ）监测大脑中动脉血流速率（Ｖ－ＭＣＡ），观察不同剂量异丙酚对脑血流动力学的影响。方法：２４例神经外科病人随机分成三组（每组８例），异丙酚剂量分别为０．２５ｍｇ／ｋｇ、１．０ｍｇ／ｋｇ和２．０ｍｇ／ｋｇ采用经颅多普勒监测双侧大脑中动脉血流速率（Ｖ－ＭＣＡ），同时监测ＢＰ、ＨＲ、ＰＥＴＣＯ２和ＳｐＯ２。分别于给药前、给药后５，１０和１５分钟测定。结果：异丙酚三组剂量均不同程     

小肠移植急性排斥反应期移植肠白细胞介素2受体和细胞间粘？…

目的 探讨移植肠白细胞介素２受体（ＩＬ－２Ｒ）和细胞间粘附分子１（ＩＣＭＡ－１）的表达在小肠移植排斥反应中的意义及诊断价值．方法 选用近交系大鼠Ｆ３４４／Ｎ和Ｗｉｓｔａｒ／Ａ进行全小肠异位移植,实验分４组,第１组：Ｗｉｓｔａｒ;第２组：Ｗｉｓｔａｒ→Ｗｉｓｔａｒ;第３组：Ｆ３４４→Ｗｉｓｔａｒ;第４组：Ｆ３４４→Ｗｉｓｔａｒ＋环孢霉素Ａ（６ｍｇ·ｋｇ＾－１·ｄ＾－１）。术后第３、５、７天取各组动物     

环孢素A及硫唑嘌呤对大鼠肝毒性的对比研究

给Ｗｉｓｔａｒ大鼠分别胃饲环孢素Ａ（ＣｓＡ）５０ｍｇ·ｋｇ~（－１）·ｄ~（－１），硫唑嘌呤（Ａｚａ）１０ｍｇ·ｋｇ~（－１）·ｄ~（－１），ＣｓＡ３０ｍｇ·ｋｇ~（－１）·ｄ~（－１）加Ａｚａ５ｍｇ·ｋｇ~（－１）·ｄ~（－１）及橄榄油２周，检测肝功能指标，血清丙二醛（ＭＤＡ），全血谷胱甘肽（ＧＳＨ）含量及肝组织病理学变化。结果Ａｚａ组血清ＡＬＴ、ＴＢｉｌ、ＭＤＡ升高以及肝组织损害最明显，ＣｓＡ组血清ＴＰ、ＡＬｂ降低、ＡＫＰ升高较Ａｚａ组及合并用药组更明显。结果表明ＣｓＡ与Ａｚａ导致肝损害的机理不同，Ａｚａ肝毒性大于ＣｓＡ，且为剂量依赖性。     

免疫抑制方案对肾移植并发慢性病毒性肝炎预后的影响

１９９１至１９９４年４月中山医科大学附一医院共完成１０１例肾移植。随访时间２８．６±９．６个月。术前供受者淋巴细胞毒性试验阴性。术中及术后２天，每日用甲基泼尼松龙（ＭＰ）０．２５～０．５ｇ，同时选用二联或三联免疫抑制方案。三联方案为泼尼松（Ｐｒｅｄ）、硫唑嘌呤（Ａｚａ）加环孢素Ａ（ＣｓＡ）；Ａｚａ于术前１天开始用１ｍｇ·ｋｇ－１·ｄ－１；Ｐｒｅｄ于术后第３天开始３０ｍｇ／ｄ，３个月后开始减量，一年后减为１０ｍｇ／ｄ；ＣｓＡ一般术后第３天开始５～８ｍｇ·ｋｇ－１·ｄ－１。二联方案为Ｐｒｅｄ加ＣｓＡ…     

激素性股骨头坏死病程中骨形态发生蛋白—2的改变 …

目的 观察激素性股骨头坏死病程中骨形态发生蛋白－２（ＢＭＰ２）的变化。方法 用醋酸氢化泼尼松诱发早期股骨头坏死动物模型,实验动物根据醋酸氢化泼尼松用量分为Ａ组（对照组）,Ｂ组（４ｍｇ／ｋｇ体重）,Ｃ组（８ｍｇ／ｋｇ体重）,Ｄ组（１６ｍｇ／ｋｇ体重）,肌注给药,每周１次。取股（肱）骨头标本行免疫组织化学染色。采用图像分析测得平均染色面积百分比和平均吸光度。结果 Ａ、Ｂ、Ｃ,Ｄ组阳性染色区染色强度依次     

异丙酚在成人体外循环中抗氧化效应的研究

目的 探讨异丙酚在体外循环（ＣＰＢ）心脏直视手术中的抗氧化效能。方法 选择在ＣＰＢ下行心脏直视手术的成年病人３０ 。主动脉阻断后,Ａ组和Ｂ组病人分别给予异丙酚０．１ｍｇ．ｋｇ＾－１．ｍｉｎ＾－１和芬太尼５ｕｇ．ｋｇ＾－１．ｍｉｎ＾－１维持麻醉。两组病人分别于麻醉前、ＣＰＢ前、ＣＰＢ３０ｍｉｎ、ＣＰＢ结束时、ＣＰＢ后１ｈ、手术结束时、以及术后１２和２４ｈ取动脉血测定红细胞中－６磷酸葡萄糖脱氢酶（Ｇ－     

中华眼镜蛇蛇毒因子在豚鼠—大鼠异种心脏移植中的作用

目的 应用中眼镜蛇蛇毒因子（CVF）消耗补体,观察豚鼠心脏在移植入Wistar大鼠腹腔内后对超急性排斥反应的变化。方法 按0．2μg/g体重CVF大鼠腹腔内分两次间隔6小时注射,18小时后进行心脏移植。脾切除及腹腔内注射环磷酰胺（Cy）均在移植前一天进行。设计分为四组：A组为对照组,不用任何药物;B组仅用CVF;C组应用CVF＋Cy＋脾切除;D组Cy＋脾切除。Cy的用量为60mg/kg体重腹腔内注射。检测各组受体的供心存活时间,并在供心停跳后取出行光镜、电镜检查。结果 A、B、C、D各组供心存活时间分别为15－3120分钟。供心存活时间的统计学分析：A组与B、C两组比较P值＜0．01,A组与D组比较,B组与C组比较P值＞0．05,B组与D组比较,C组与D组比较P值＜0．01。光镜、电镜结果提示：B、C组与A、D组有明显不同。结论 CVF能明显抑制补体活性,减轻或延缓超急性排斥反应的发生,使供体器官存活时间延长。CVF具有异种器官移植的基础研究和临床开发意义。     

Inhibition of the membrane attack complex of complement for induction of accommodation in the hamster-to-rat heart transplant model

BACKGROUND: To induce accommodation in the hamster-to-rat cardiac transplantation model, in addition to cyclosporin A (CSA) to inhibit T-cell-mediated graft rejection, cobra venom factor (CVF) is often used to prevent complement-mediated graft rejection. Although it is generally assumed that CVF makes accommodation possible because it inactivates the complement membrane attack complex (MAC), it is not known which complement components must be inactivated and whether complement activation products generated by CVF are also involved in the induction of accommodation. Therefore, to investigate mechanisms by which CVF contributes to accommodation, we studied induction of accommodation of hamster hearts grafted into rats with complement deficiencies of C6; these rats cannot assemble the MAC but, in contrast to CVF, retain in their native state all complement proteins that precede the MAC. METHODS: Golden Syrian hamster hearts were transplanted heterotopically into the abdomen of normocomplementemic and C6-deficient (C6D) PVG rats. Graft rejection was determined by cessation of palpable cardiac contractions. CSA, 10 mg/kg, was administered daily to all rats. Graft survival was compared in rats given CVF (60 U/kg 1-day pre-transplant and 20 U/kg/day for the next 9 days), C6D rats given no CVF, normocomplementemic rats given anti-C6 IgG or non-immune IgG but no CVF, and C6D rats reconstituted with normocomplementemic rat serum. Total complement and C6 serum levels were measured using hemolytic assays in rat peripheral blood. RESULTS: We found that hamster hearts transplanted into C6D rats receiving CSA but no CVF survived long-term, with histology typical of an accommodated heart. The accommodated hamster heart did not reconstitute C6 levels of the C6D recipient rats. Moreover, in normocomplementemic rats given anti-C6 antibodies (abs) to induce partial C6 deficiency, accommodation also developed without administration of CVF. Accommodation of the hamster heart failed to develop in C6D rats whose complement was reconstituted by administration of normocomplementemic rat serum given before and following transplantation. CONCLUSIONS: These studies demonstrate that, in this model, inhibition of MAC-mediated graft rejection is sufficient to allow the development of accommodation. Inactivation of C3 or other complement proteins of the alternate pathway, or the presence of complement-derived biologically active fragments, is not needed for development of accommodation.     

A highly selective inhibitor of Rho-associated coiled-coil forming protein kinase, Y-27632, prolongs cardiac allograft survival of the BALB/c-to-C3H/He mouse model.

BACKGROUND: Current studies provide evidence that a small G protein, RhoAp21, and its target protein, Rho-associated coiled-coil forming protein kinase (ROCK), regulate not only cell shape but also cell migration. However, contribution of Rho/ROCK signaling to graft rejection is unknown. The purpose of this study was to evaluate the inhibitory effect of Y-27632, a highly selective ROCK inhibitor, on rejection of heterotopic cardiac transplantation in mice. METHODS: BALB/c (H-2(d)) hearts were transplanted into C3H/He (H-2(k)) as allografts that were full histoincompatibility combinations. The recipients received several doses of Y-27632, commencing 1 day before cardiac transplantation until rejection. We used immunohistochemical study to detect the expression of myocardial intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1), and we immunoenzymatically measured serum interleukin (IL)-6. Furthermore, we evaluated cardiac allograft vasculopathy treated with either FK506 or Y-27632 at Day 100. RESULTS: The Y-27632-treated (2 mg/kg/day) allografts prolonged the mean survival time (49.6 +/- 10.1 days, n = 12) as compared with the untreated allografts (8.1 +/- 0.4 days, n = 7, p < 0.001). Histologic examinations of the Y-27632-treated allografts at Day 7 showed greatly reduced leukocyte infiltration compared with the untreated allografts. The Y-27632-treated allografts revealed faint expression of myocardial ICAM-1 and VCAM-1 at Day 7. The serum IL-6 levels also decreased in the Y-27632-treated mice. In the long-surviving Y-27632-treated allografts at Day 100, we saw neither active rejection nor apparent thickening of vascular intima. CONCLUSION: Our results suggest that ROCK plays a major role in cardiac rejection in the BALB/c-to-C3H/He mouse model. Inhibition of this Rho/ROCK signaling may be an alternative therapeutic option for managing acute and chronic rejection.     

Prolonged Cardiac Allograft Survival in Mouse Model After Complement Depletion with Yunnan Cobra Venom Factor

Background

Activation of the complement system is the leading mechanism that causes antibody-mediated acute rejection and hyperacute rejection after xenotransplantation. The major cause of acute rejection in allogeneic transplantation is the T cell–mediated specific immune response. We studied the effects of complement on acute rejection after cardiac allotransplantation using complement depletion with cobra venom factor (CVF) in the mouse.

Materials and Methods

The Balb/c-C57 mouse model of heterotopic cardiac allograft was used. The mice were divided into 2 groups, a control group and a CVF-treated group. After intravenous injection of CVF, the experimental group was observed for allograft survival time. Twelve mice from the control and experimental groups were sacrificed on days 3, 5, and 7 after the operation. The pathologic grade of acute rejection, deposition of C3 in tissue, extent of infiltration by CD4+ and CD8+ T cells, and expression of MHC-II, B7-1, and B7-2 were compared between the 2 groups.

Results

In the CVF-treated group, mean (SD) survival of the cardiac allograft was 26.2 (1.7) days, and in the control group was 8.4 (0.4) days (P < .01). Pathologic examination and immunohistochemistry demonstrated that the grade of acute rejection, deposition of C3 in tissue, extent of infiltration of CD4+ and CD8+ T cells, and expression of MHC-II, B7-1, and B7-2 were significantly decreased in the CVF-treated group.

Conclusion

Depletion of complement in the serum with CVF inhibits acute cardiac allograft rejection in the mouse.     

Prolongation of discordant renal xenograft survival by depletion of complement. Comparative effects of systemically administered cobra venom factor and soluble complement receptor type 1 in a guinea-pig to rat model

Abstract There is an increasing body of evidence to suggest that inhibition of complement activation may be a valuable approach to avert hyperacute rejection. In our study, the guinea-pig to rat discordant kidney xenograft model was adapted for the investigation of renal transplant function and an attempt was made to delay the hyperacute rejection using systemically administered cobra venom factor (CVF) and soluble complement receptor type 1 (sCR1). The saline-treated control recipients experienced a rapid transplant rejection with a xenograft survival averaging 10.5 ± 2.1 min. Administration of a single 60 U/kg i. v. bolus of CVF significantly prolonged renal graft survival to 20.4 ± 2.5 h, and by a single bolus of sCR1 (50 mg/kg) a prolongation of graft survival to 18.8 ± 2.3 h was achieved. The grafts functioned only over periods of 2.5 ± 0.3 and 2.3 ± 0.2 h, respectively. No complications of sCR 1 were noted. We concluded that complement inhibition by sCR 1 may be an important component in the therapeutic approach aiming at the prevention of hyperacute rejection in human organ transplantation.     

Effect of LS-2616 on the graft protection achieved by cyclosporin A,prednisolone, and 15-deoxyspergualin in heart-transplanted rats

The immunostimulator LS-2616 abolishes the effect of cyclosporin A in a rat cardiac transplantation model. The present paper compares the characteristics of rejection obtained under different immunosuppressive regimens with and without additional LS-2616 application in the same model. Cyclosporin A (CyA, 10 mg/kg daily), prednisolone (15 mg/kg daily), or 15-deoxyspergualin (2, 5, or 10 mg/kg daily), all given from the day of transplantation until day 9, protected the grafts during the treatment period. The addition of LS-2616 (160 mg/kg, day —1 until stop) resulted in a total abrogation of the immunosuppressive effect of CyA and prednisolone. However, LS-2616 could only partially or not at all reverse the effect of 15-deoxyspergualine. These results show a certain drug selectivity of LS-2616 in promoting rejection of immunosuppressed allografts. Further studies with LS-2616 may be of benefit in evaluating the mode of action of different immunosuppressive compounds and, thus, contribute to finding more effective antirejection therapies.     

Beneficial effects of pentoxifylline pretreatment in non-heart-beating donors in rats

BACKGROUND: Pentoxifylline (PTX) pretreatment of recipients was shown to protect against liver graft failure from ischemia-reperfusion injury after orthotopic rat liver transplantation. It has also been shown that PTX protects against normothermic ischemia-reperfusion injury to the liver in lobar ischemia model in the rat. Whether PTX can benefit the liver procured from non-heart-beating donors (NHBDs) with up to 9 hr of cold ischemia is unknown. METHODS: Donor and recipient rats were pretreated with intraperitoneal PTX (50 mg/kg) 1 hr before cardiac arrest and transplantation, respectively. Grafts were transplanted 0, 30, and 60 min after cardiac arrest with additional 1 and 9 hr of cold ischemia in both PTX-pretreated or untreated (control) groups (10 rats per group). PTX (25 mg/kg/day) was continuously given to the surviving rats for 5 days postoperatively. Recipient survival rates, serum enzyme levels, and histopathological examination of postreperfusion liver biopsies were all analyzed. RESULTS: The survival rates, serum enzyme levels, and postreperfusion histology were significantly improved in groups pretreated with PTX compared to the controls. CONCLUSION: Donor and recipient PTX pretreatment significantly improves the viability of the liver grafts procured from NHBDs.     

Macrophage depletion prevents anti-graft antibody production and results in long-term survival in xenotransplantation

Liposome-encapsulated dichloromethylene diphosphonate (clodronate) is known to deplete macrophages. We examined the effect of clodronate on xenoreactive antibody production and xenograft rejection. Hamster cardiac grafts were transplanted into Lewis rats. Clodronate (4 mL/kg) was injected intravenously on the day before transplantation. In some groups, cyclosporine A (CsA) at a dose of 15 mg/kg was given daily intramuscularly until the end of each experiment. Untreated Lewis rats rejected the grafts at 2 and 3 days after transplantation. Neither CsA treatment alone nor clodronate treatment alone prolonged graft survival. Five of 7 Lewis recipients treated with clodronate and CsA did not reject hamster hearts for 100 days. Antibody production in the CsA plus clodronate-treated group was suppressed compared with control groups.     

Increased expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in rat cardiac allografts

This study was designed to investigate the role of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) during chronic cardiac allograft rejection. Wistar rats were used as donors, and SD rats as recipients heterotopic cardiac transplants. Recipients pretreated with inoculation of donor splenocytes (SPC) followed by cyclophosphamide (CP) were divided into 4 groups: (A) untreated group (n = 18) without immunosuppression; (B) SPC plus CP-treated group (n = 18) that were euthanized at 15-120 days posttransplantation; (C) CsA-treated group (n = 18) euthanized at 2-3 months posttransplantation; and (D) tolerance group (n = 18) treated with SPC plus CP and monitored for at least 1 year posttransplantation. Cardiac allografts were harvested at various times for immunohistochemical studies performed to evaluate the expression of ICAM-1 and VCAM-1. Pretreatment of animals with SPC and CP induced long-term cardiac allograft survival. Immunohistochemical staining demonstrated a low level of ICAM-1 and VCAM-1 expression in cardiac allograft muscle and coronary arteries among Groups B and D. In contrast, the expressions of ICAM-1 and VCAM-1 in cardiac allografts of Groups A and C were significantly higher than those in Groups B and D. Our results suggested that the expression of ICAM-1 and VCAM-1 plays an important role during the development of chronic cardiac allograft rejection.