Isolated Pelvic Hyperthermochemotherapeutic Perfusion-An Experimental Study on Isolating Efficacy

Hyperthermochemotherapeutic perfusion model through isolated pelvic vessels was developed to evaluate the leakage of hyperthermia and drugs (such as adriamycin) from the isolated pelvic circulation to systemic circulation and its associated side/toxic effects. The isolated pelvic circulation was perfused through a femoral artery catheter with hyperthermic (48 ℃ to 55 ℃) adriamycin solution (50 μg/ml) for 30 min. The efflux was drained through a femoral vein catheter. And the pelvic temperature was kept at the level of 43±0.5 ℃. The temperature of pelvic circulation was kept at 4 ℃ to 5 ℃ greater than the systemic/core temperature. The adriamycin concentration of pelvic efflux was 12 to 46 folds of that of systemic serum. The difference between them was very significant ( P <0.001). As the perfusion pressure was increased, which kept lower than the mean systemic artery pressure, the leakage of the adriamycin from the isolated pelvic circulation to systemic circulation was increased, but there was no significant difference between them ( P >0.05). During isolated perfusion, the systemic blood dynamics remained stable and there were no organic injuries on the important organs. It was suggested that the isolating efficacy of the modality of isolated pelvic hyperthermochemotherapeutic perfusion through vessels was rather high. The hyperthermia and drugs could be effectively limited in the isolated pelvic region with minor side effects on the systemic circulation and important organs.

Isolated hepatic perfusion: a regional therapy for liver cancer

BACKGROUND: Many treatments have been proposed for non-resectable primary or secondary hepatic cancer but the results have been disappointing. Isolated hepatic perfusion (IHP) was attempted five decades ago but it has been ac- cepted recently after spectacular tumour responses were ob- tained by several phase trials. DATA SOURCES: An English-language literature search using MEDLINE (2003), Index Medicus (2003) and biblio- graphic reviews of books and review articles. IHP and its history and recent clinical application. RESULTS: IHP offers unique pharmacokinetic advantages for locoregional chemotherapy and biotherapy. Surgical isolation of the liver and percutaneous techniques using bal- loon occlusion catheters are reliable and safe. They appear to have significant efficacy even in patients with advanced tumor burden or those with tumors refractory to other types of therapy. CONCLUSION: IHP which has been developed in recent years is becoming a promising strategy for the treatment of unresectable liver cancer.

Isolated hepatic perfusion for unresectable hepatic malignancies:A systematic review and meta-analysis

AIMTo investigate the effcacy and safety of isolated hepatic perfusion （IHP） in the management of unresectable liver malignancies.METHODSStudies were identified manually and on-line by using PubMed and EMBASE database. We formulate the eligibility criteria according to the PICOS elements, and accessed the quality of studies using the MINORS instrument. Data from all included studies were carefully investigated. We calculated the pooled response rate and incidences of mortality reported from all eligible studiesby using the Meta-Analyst software, and we computed a pooled relative risk （RR） and 95%CI by using the Comprehensive Meta-Analysis software. Heterogeneity was quantifed evaluated using I2 statistic.RESULTS Eight studies, including 502 patients, were selected. Of these, six studies performed IHP, while the other two studies performed percutaneous IHP. The results showed that the pooled response rate was 60.8% （95%CI： 53.1%-68%）, I2 = 37.1%. The median overall survival was 20 mo （range： 12.1 to 25 mo） following IHP or PIHP. The pooled mortality rate was 5.4% （95%CI： 2.5%-11.2%）, I2 = 37.5%. Prognostic factors predict the response to IHP or survival, and were reported in six studies. Meta-analysis demonstrated that Gender was not associated with overall survival （RR = 0.877, 95%CI： 0.564-1.365）; however, carcino-embryonic antigen ≤ 30 ng/mL was associated with a signifcant improvement in survival outcomes with colorectal cancer patients （RR = 2.082, 95%CI： 1.371-3.163）, and there was no signifcant heterogeneity.CONCLUSIONThe present systemic review and meta-analysis suggest that IHP and PIHP are potentially efficient and safe techniques for unresectable liver primary and secondarymalignancies.

Anatomical basis for pancreas transplantation via isolated splenic artery perfusion:A literature review

The variability of vascular anatomy of the pancreas underlines the difficulty of its transplantation.Research regarding the consistency of anatomical variations shows splenic arterial dominance in most cases.This can significantly improve transplantation success.A systematic literature review was performed according to the quality standards described in the AMSTAR measurement tool and the PRISMA guidelines.We valuated existing literature regarding the vascularization and blood perfusion patterns of the pancreas in terms of dominance and variability.The collected data was independently analyzed by two researchers.Variance of vascular anatomy was seen to be underreported in literature,though significant findings have been included and discussed in this study,providing valuable insight into the dynamics of pancreatic perfusion and feasibility of transplantation on several different supplying arteries.The splenic artery(SA)has a high percentage of consistency in all found studies(over 90%).High frequency of anastomoses between arterial pools supplying the pancreas can mediate sufficient blood supply through a dominant vessel,such as the SA,which is present in most cases.Pancreatic transplantation with isolated SA blood supply can provide sufficient arterial perfusion of the pancreas for stable transplant viability due to high anatomical consistency of the SA and vast communications with other arterial systems.

Glycyrrhizinate reduces portal hypertension in isolated perfused rat livers with chronic hepatitis

AIM:To investigate the effects of diammonium glycyrrhizinate(Gly)on portal hypertension(PHT)in isolated portal perfused rat liver(IPPRL)with carbon tetrachloride(CCl4)-induced chronic hepatitis.METHODS:PHT model was replicated with CCl4 in rats for 84 d.Model was identified by measuring the ascetic amounts,hepatic function,portal pressure in vivo,splenic index,and pathological alterations.Inducible nitric oxide synthase(iNOS)in liver was assessed by immunohistochemistry.IPPRLs were performed at d0,d28,d56,and d84.After phenylephrine-induced constriction,Gly was geometrically used to reduce PHT.Gly action was expressed as median effective concentration(EC50)and area under the curve(AUC).Underlying mechanism was exploited by linear correlation between AUC values of Gly and existed iNOS in portal triads.RESULTS:PHT model was confirmed with ascites,splenomegaly,serum biomarkers of hepatic injury,and elevated portal pressure.Pathological findings had shown normal hepatic structure at d0,degenerations at d28,fibrosis at d56,cirrhosis at d84in PHT rats.Pseudo lobule ratios decreased and collagen ratios increased progressively along with PHT development.Gly does dose-dependently reduce PHT in IPPRLs with CCl4-induced chronic hepatitis.Gly potencies were increased gradually along with PHT development,characterized with its EC50at 2.80×10-10,3.03×10-11,3.77×10-11and 4.65×10-11mol/L at d0,d28,d56and d84,respectively.Existed iNOS was located at hepatocyte at d0,stellate cells at d28,stellate cells and macrophages at d56,and macrophages in portal triads at d84.Macrophages infiltrated more into portal triads and expressed more iNOS along with PHT development.AUC values of Gly were positively correlated with existed iNOS levels in portal triads.CONCLUSION:Gly reduces indirectly PHT in IPPRL with CCl4-induced chronic hepatitis.The underlying mechanisms may relate to rescue NO bioavailability from macrophage-derived peroxynitrite in portal triads.

Effects of amniotic fluid embolism-like plasma on isolated perfused rabbit lungs

<Abstract>Effects of amniotic fluid embolism-like plasma (AFEP) on the isolated perfused rabbit lungs (IPRL)were studied. It was found that AFEP could induce elevation of pulmonary artery pressure (PAP) and development of lung edema, which could be partially prevented by ibuprofen, a cycloxygenase inhibit0r, but amniotic fluid itself could not cause elevation of PAP and lung edema. The result suggests that AFEP-induced mediator from whole blood cells may be the important factor resulting in above-mentioned pathological changes.

NMR Study of Damage on Isolated Perfused Rat Heart Exposed to Ischemia and Hypoxia

Myocardial ischemia is the most common and primary cause of myocardium damage. Numerous conventional techniques and methods have been developed for ischemia and reperfusion studies. However, because of damage to the heart sample, most of these techniques can not be used to continuously monitor the full dynamic course of the myocardial metabolic pathway. The nuclear magnetic resonnance (NMR) surface coil technique, which overcomes the limitations of conventional instrumentation, can be used to quantitatively study every stage of the perfused heart (especially after perfusion stoppage) continuously, dynamically, and without damage under normal or designed physiological conditions at the molecular level. In this paper, 31 P NMR was used to study the effects of ischemia and hypoxia on isolated perfused hearts. The results show that complete hypoxia caused more severe functional damage to the myocardial cells than complete ischemia.

丹参素对大鼠离体心脏缺血再灌注心肌能量代谢的影响

SD大鼠随机分为正常灌流组和缺血再灌注组,对大鼠离体心脏进行Langendorff灌流。采用高效液相色谱法测定大鼠心肌组织中辅酶A、乙酰辅酶A及高能磷酸化合物含量,心肌组织中乳酸含量和灌流液中脂肪酸的含量采用试剂盒测定,观察给予丹参素和阳性药雷诺嗪后上述各指标的变化情况。结果表明,雷诺嗪能够增加心肌ATP储备,升高腺苷酸能荷值,降低心肌组织乳酸含量及含脂肪酸组乙酰CoA/CoA比值,抑制脂肪酸的氧化从而优化心肌能量代谢。丹参素可以使大鼠离体心脏ATP含量及腺苷酸能荷值升高,乳酸含量降低,对正常灌流组心肌组织乙酰CoA/CoA比值无明显影响,但可以降低添加脂肪酸的缺血再灌注组心肌组织乙酰CoA/CoA比值,提示丹参素对病理状态下心肌脂肪酸的氧化有一定的抑制作用,从而优化病理状态下的心肌能量代谢,发挥对缺血再灌注心肌的保护作用。

丹参注射液对离体大鼠心肌能量代谢的调节作用

目的:研究丹参注射液对大鼠离体心脏能量代谢的调节作用。方法:采用大鼠离体心脏灌流技术,测定在添加或不添加脂肪酸的情况下,大鼠心脏中乙酰辅酶A(AcCoA)、辅酶A(CoA)、高能磷酸化合物、乳酸的含量,并观察给予丹参注射液和阳性药肝乐(DCA)干预后各指标的变化。结果:在不含脂肪酸的情况下,DCA和丹参注射液均显著提高大鼠心肌组织中AcCoA/CoA值;在含有脂肪酸的情况下,DCA对AcCoA/CoA值没有显著影响,而丹参注射液显著降低了AcCoA/CoA值;在含有脂肪酸的情况下,加入丹参注射液会导致DCA组大鼠心肌组织中AcCoA/CoA值降低。丹参注射液可使离体大鼠心脏中ATP含量升高,能荷值升高,乳酸含量降低。结论:丹参注射液抑制离体大鼠心脏中脂肪酸的氧化,提高心肌能量代谢的效率,起到心肌保护作用。

尾加压素Ⅱ对正常及缺血-再灌注离体大鼠心脏的影响

在正常Langendorff灌流与缺血-再灌注(停灌20 min-复灌20 min)离体大鼠心脏模型,观察尾加压素Ⅱ(urotensin Ⅱ,UⅡ)对冠脉流量、心功能和心肌代谢的影响以及心肌UⅡ受体的功能,以探讨UⅡ的心脏效应。对正常心脏给予0.1、1和10 nmol/L UⅡ各5 min,然后换洗5 min,对停灌缺血-再灌注心脏在再灌注期给予1或10nmol/L UⅡ。监测心率、左室内压和左室内压升降的最大变化率等心功能指标,计算冠脉流量,测定冠脉流出液中总蛋白和肌红蛋白含量以及乳酸脱氢酶(LDH)活性。灌流结束后,测定心肌丙二醛(MDA)含量和质膜UⅡ结合位点(放射性配基结合法)。结果如下:(1)正常心脏灌流UⅡ后,冠脉流量和心功能呈浓度依赖下降,换洗后没有完全恢复。心肌蛋白、肌红蛋白和LDH漏出随UⅡ浓度的增加而增加,换洗后迅速减少。UⅡ组心肌MDA含量与对照组差异无显著性。(2)缺血-再灌注后,冠脉流量显著减少,心功能显著抑制,再灌注期心肌蛋白、肌红蛋白和LDH明显漏出;给予UⅡ后,上述变化增强,且高浓度组更强,与对照组差异有显著性(P<<0.01),再灌注后心肌MDA含量亦显著高于对照(P<0.01)。(3)缺血-再灌注心肌质膜UⅡ受体的B_(max)显著高于正常对照心肌(14.65±1.78vs20.53±1.98 fmol/mg pr,P<0.01),Kd值变化无统计学意义。上述结果表明,在正常?

原位循环灌流法分离大鼠肝星状细胞

目的：为深入探讨肝纤维化的细胞机制，探索经济、稳定的肝星状细胞分离和培养方法。方法：采用胶原酶原位循环灌流法分离大鼠肝星状细胞，１８％Ｎｙｃｏｄｅｎｚ密度梯度离心纯化星状细胞。结果：星状细胞的得率为１×１０７～３×１０７／肝，纯度为９０％，存活率为９６％。结论：原位循环灌流法是有效的大鼠肝星状细胞的分离和培养方法。

隔离肝灌注动物实验

隔离肝灌治疗方法可以提高治疗肝癌的化疗药物局部浓度，减轻全身的毒副作用，由于手术难度大、手术操作复杂，手术技术的基本训练成为临床应用前的重要手段。为了达到熟练操作的目的，须完成大量的动物手术训练。该课题组选用大鼠作为训练材料，作为大动物手术训练前的基础训练。该文分析了大鼠手术训练死亡的原因，记录了手术中和灌注过程中大鼠的生理、病理反应，介绍了取消循环灌注泵进行手术新的方法。对５８只大鼠隔离肝灌注的手术生存率和手术并发症进行了分析。结果发现：手术操作技术、手术路径和手术器械的改进，使手术生存率大大提高，手术并发症明显减少。

用于核磁共振检测的离体心脏灌流模型

本报告是关于核磁共振研究生理学问题的灌流模型。我们解决了Langendorff灌流装置用于核磁共振研究时存在的问题，如远距离灌流、保温、保氧和灌流液回收等，建立了稳定的可供核磁共振测量用的灌流模型。在这个装置中高体大鼠心脏的节律性活动可维持90min以上。

经皮选择性肝脏隔离灌注化疗方法学研究

目的 探讨经皮选择性隔离肝脏灌注化疗的可行性及隔离效果。方法 将 12只猪随机分为两组 :HAI组 6只 ,进行常规经肝动脉灌注化疗 ;PSIHP组 6只 ,利用介入放射学方法进行经皮选择性隔离肝脏灌注化疗结合血液灌流。化疗药物选用 5 -FU ,分别检测肝静脉及外周血液中的血药浓度。结果 HAI组肝静脉血 (过滤前 )和外周静脉血浓度峰值分别为 (4 15 3 4± 4 30 2 ) ,(16 76 1± 2 2 6 9)mg/L ;PSIHP组分别为 (5 2 19 7± 5 30 2 ) ,(6 5 7± 6 4 )mg/L。两组差异有显著性 (P <0 0 1)。PSIHP组过滤后肝静脉血血药浓度峰值为 (4 0 8 7± 4 7 8)mg/L ,碳肾率过滤达到 (78 2± 7 4 ) %。结论 PSIHP是一种简单有效的肝脏隔离灌注化疗技术 ,与常规经肝动脉灌注化疗相比 ,不仅可以增加局部血药浓度 ,更可以降低外周血血药浓度 。

高温隔离灌注化疗在下肢骨与软组织肉瘤保肢治疗中的价值与限度

为了减低肢体恶性肿瘤治疗中保肢手术的局部复发率，我们从１９９０年开始对５７例下肢ⅡＢ期骨与软组织肉瘤进行术前顺铂或卡铂高温（４１～４３°Ｃ）隔离灌注化疗，同时结合静脉滴注阿霉素。本组进行保肢手术４７例、截肢手术１０例。术前化疗后显示９１％患者局部疼痛完全消失；９５％肿块明显缩小；８９．５％关节活动度改善。术后病理检查肿瘤坏死率９０％～１００％为完全反应者４９例（８６％）；６０％～９０％为部分反应者５例（８．８％）；６０％以下为无反应者３例（５．２％）。术后随访２～５年的２６例保肢者中局部复发２例（７．７％）；术后３年以上随访者２６例，无病存活１６例（６１．５％）。作者认为本法局部效果非常良好，十分利于施行保肢术手，但无助于提高存活率，对截肢者不必应用。

丹参水溶性成分含量变化对大鼠离体心肌能量代谢调节的影响

通过离体心肌缺血/再灌注模型,考察丹参注射液各水溶性成分含量变化对大鼠离体心肌能量代谢调节的影响。大鼠随机分为模型对照组、丹参注射液组和4个成分外加组。大鼠离体心脏发生缺血/再灌注时给予相应药物,测定心肌组织中乙酰辅酶A(AcCoA)、辅酶A(CoA)、高能磷酸化合物及心肌缺血损伤标记物[乳酸(LD)、乳酸脱氢酶(LDH)和肌酸激酶(CK)]等药效指标。与对照组相比,外加原儿茶酸(PA)组显著降低AcCoA/CoA比值(P<0.05),显著升高总腺苷(TAN)和腺苷酸能荷(AEC)(P<0.05);外加丹酚酸B(SalB)组所造成的影响与外加PA类似,但对能量代谢指标的调节作用总体弱于PA。对实验数据建立反向传播神经网络模型,采用Garson算法计算4种水溶性成分对药效指标的贡献率。各成分对脂肪酸代谢类和高能磷酸化合物类指标的影响排序从大到小依次均为:PA、SalB、SalA、DSS;心肌缺血损伤标记物受各成分含量的影响顺序从大到小依次为:PA、SalB、DSS、SalA。结果提示,丹参注射液水溶性成分含量的改变能增加心肌ATP储备,升高腺苷酸能荷值,同时降低心肌组织中AcCoA/CoA比值,从而提高心肌能量代谢效率,这种作用尤以丹参注射液中酚酸类物质最为显著。

提高离体蛙心灌流实验成功率的方法研究

离体蛙心灌流实验是生理学中一个重要的实验项目。近年来的教学实践表明,插管过程导致蛙心受损、插管内血液凝固堵塞、动脉切口位置不当等是易导致该实验失败的主要原因。选择合适的实验动物及插管,以及插管方法的改进、控制灌流液温度等措施可有效提高离体蛙心灌流实验的成功率。

离体大鼠肾脏灌流技术在发现和评价促尿酸排泄药物中的初步应用

目的为建立发现和评价促尿酸排泄药物作用的离体模型,采用离体大鼠肾脏灌流技术,观察尿酸在离体肾脏中排泄特点和丙磺舒对尿酸排泄影响。方法制备离体大鼠肾脏灌流模型,随机分为对照组和丙磺舒处理组,以含复方氨基酸和65g.L-1牛血清白蛋白的K-H灌流液循环灌流90min,定时收集导尿管流出液和灌流液,测定其中菊粉、钠、葡萄糖及尿酸含量,计算肾功能参数和尿酸排泄情况。结果离体灌流肾脏的肾小球滤过率,尿量,肾小管重吸收水、钠、葡萄糖百分率等肾功能参数在灌流的90min期间处于稳定的生理范围内;对照组灌流肾脏尿酸排泄分数、尿酸清除率及90min内总尿酸排泄量分别为30%、0.30ml.L-1和15μmol,与对照组比较,丙磺舒使离体肾脏尿酸排泄分别提高36%、54%和99%。结论离体大鼠肾脏灌流技术可用于发现和评价促尿酸排泄药物作用。

6,7-二甲氧基香豆素抗急性肾功能衰竭的作用机制研究

目的 探讨６ ，７二甲氧基香豆素抗急性肾功能衰竭（ Ａ Ｒ Ｆ） 的作用，并初步分析其作用机制。方法 观察６ ，７二甲氧基香豆素对内毒素诱导小鼠 Ａ Ｒ Ｆ 模型的影响，分析其对 Ａ Ｒ Ｆ 的作用；观察消炎痛或 Ｌ Ｎ Ａ Ｍ Ｅ 协同 Ｎ Ｅ 升高离体灌流肾肾灌注压以分析６ ，７二甲氧基香豆素对 Ｐ Ｇ Ｉ２ 和 Ｎ Ｏ的影响。结果 ６ ，７二甲氧基香豆素对内毒素引起的小鼠急性肾衰有明显的拮抗作用，能降低血清肌酐浓度、血清尿素氮浓度、肾匀浆 Ｓ Ｏ Ｄ 活性及肾匀浆 Ｌ Ｐ Ｏ 含量的升高，减轻肾小球毛细血管及肾间质充血。６ ，７二甲氧基香豆素有延缓正常灌流肾肾灌注压缓慢上升的作用。呈剂量依赖性拮抗 Ｎ Ｅ 引起的肾灌注压的升高。通过 Ｐ Ｇ Ｉ２ 及 Ｎ Ｏ 的释放等减小肾血管阻力。结论 ６ ，７二甲氧基香豆素有抗内毒素诱导的 Ａ Ｒ Ｆ 的作用，主要通过防止氧自由基的生成及（ 或） 清除已生成的氧自由基，促使血管内皮细胞释放 Ｎ Ｏ 和 Ｐ Ｇ Ｉ２ 。

成功制作离体心脏灌注模型要点和经验

目的探讨并总结制作Langendorff离体心脏灌注模型的要点及成功经验。方法通过16只健康SD大鼠(体质量200～400 g),以HTK液作为心脏停搏液建立标准的Langendorff离体心脏灌注模型,将此模型与持续K-H液心肌灌注、持续心脏搏动组大鼠的同一时段的心肌酶学及血流动力学指标进行对比,使用SPSS 11.5作统计学重复测量资料的方差分析。结果 1例因操作不熟练致提取心脏时间延长,1例因灌注管插入深度过深导致主动脉瓣毁损,1例因室温过低,1例因灌注管管径过大无法插入而导致实验失败,其余12例在应用了室内加热器、不同管径灌注针、自制球囊测压管及加快操作速度后均获得实验成功。所制模型心肌酶学及血流动力学各指标与持续心脏搏动组无明显差异(P>0.05)。结论掌握制作要点及注意事项后,Langendorff离体心脏灌注模型可顺利快速地成功制备。