长征—1号多器官保存液对兔肾脏冷冻保存的连续形态学观察

自１９９２年开始研制成功长征１号多器官保存液（简称ＣＺ－１液）在完成对兔心脏及肝脏的冷冻保存后的形态学变化观察后，又做了对兔肾脏冷冻保存的形态学改变观察，并与ＵＷ液和ＨＣ－Ａ液作了比较，结果表明，冷冻保存７２小时以内ＣＺ－１液组肾脏组织光镜和电镜下形态学改变与ＵＷ组相比无明显区别，而且ＣＺ－１液组冷冻保存肾脏组织的变化以及细胞和细胞器的变性均缓于ＨＣ－Ａ组，由于ＨＣ－Ａ液已被证实能有效地保存犬肾在     

长征-1号多器官保存液效果的动物实验研究

目的　研究长征１ 号（ＣＺ１） 多器官保存液对肾脏低温保存的有效性及移植存活率的影响。　方法　（１） 采用离体肾脏灌注模型,分别检测了低温延时保存后兔肾线粒体呼吸控制率（ＲＣＲ）、Ｎａ＋Ｋ＋ＡＴＰａｓｅ 活性、皮质线粒体Ｃａ２ ＋ 、皮质ＡＴＰ等含量的变化。（２） 采用ＳＤ 大鼠肾脏移植模型,分别观察了低温延时保存４８ 、７２ 小时后再移植,大鼠存活以及移植肾功能恢复情况。　结果　（１）ＣＺ１ 液组保存７２ 小时,其生化指标肾脏线粒体呼吸控制率、皮质线粒体Ｃａ２＋ 、皮质ＡＴＰ含量的变化均优于ＵＷ 液组;（２）经ＵＷ 液和ＣＺ１ 液低温保存供肾４８ 小时,７ 天之内肾功能恢复正常。保存供肾７２ 小时,１４ 天之内肾功能基本接近正常。　结论　长征１ 号多器官保存液对肾脏低温保存效果基本类同于ＵＷ 液,且部分生化学结果优于ＵＷ 液。     

自制CZ—1液对兔心肌组织低温延时保存的生化学指标观察

研制出我国自制的多种改良的ＵＷ液，是摆在移植界的一大课题。我院于１９９２年开始研制成长征－１号多器官保存液（简称ＣＺ－１液），同时对心肌组织０、６、１２、１８、２４小时冷冻保存后的生化学指标作了观察。结果表明：与ＵＷ液比较，ＣＺ－１液０、１２、１８、２４小时冷冻保存兔心肌组织ＡＴＰ含量无明显差别。而ＣＺ－１液冷冻保存心肌线粒体Ｃａ＾２＋超载明显缓于ＵＷ液。文章认为从生化指标变化来说，我们自的ＣＺ－     

自制长征—1液对兔心脏低温延时保存的形态学观察

我际于１９９２年研制成长征－１多器官保存液，同时对心肌组织０，１８，２４，３６，４８小时冷冻保存后的形态学作了观察，并与ＵＷ液作了比较。结果表明：１．从光镜观察来看，２４小时后心肌细胞浊肿，部分横纹不清。肌纤维紊乱，有断裂。肌纤维间隙增宽。４８小时后心肌细胞散在性坏死。其ＣＺ－１液组与ＵＷ液组无明显区别。２．从电镜观察来看，１８小时后心肌细胞受到轻－中度损伤。     

亚低温下肾保存的实验研究

为了进一步提高肾移植的水平，了解肾脏在低温保存下的病理改变及生化指标的变化，利用低温可控冰箱，以ＨＣＡ液作为灌洗及保存液，对兔肾进行不同温度（４℃、０℃，－２℃、－４℃）及不同时间（７２、９６、１２０小时）的保存，经对保存肾脏进行形态及功能方面观察，认为灌洗后静脉流出液ＬＤＨ浓度及光、电镜观察是肾保存质量观察的可靠指标。同时，在－２℃以下温度保存的肾皮质细胞在镜下可见有冰晶形成引起的空泡，它对肾脏细胞及结构产生严重的损害。     

大鼠肝脏保存中不同器官保存液对肝细胞凋亡的影响

目的 研究３种目前国内常用的器官保存液对供肝细胞凋亡的影响。方法 分别用ＵＷ液、ＨＣ－Ａ液和ＷＭＯ－１号液灌洗并保存大鼠肝脏,于保存后０、１２、２４ｈ用原位末端标记法检测供肝细胞凋亡情况,并将保存２４ｈ的肝及行大鼠原位全肝移植,观察受者的３ｄ存活率。结果 ３种保存液保存的肝脏均在保存１２ｈ时出现细胞凋亡,ＨＣ－Ａ液级瑟ＷＭＯ－１号液组的凋亡指数（ＡＩ）高于ＵＷ液组（Ｐ〈０．００１）,而ＨＣ－Ａ液组     

川芎嗪对犬肾保存的影响

通过犬自体肾移植模型研究了一定浓度的川芎嗪加入ＷＭＯ－１号液对犬肾保存效果的影响。结果表明，单纯ＷＭＯ－１号液保存７２小时和加川芎嗪保存７２小时的肾行自体移植均难存活；加川芎嗪保存４８小时组术后血肌酐的平均值低于单纯ＷＭＯ－１号液保存４８小时组，且血肌酐恢复正常的时间也较单纯ＷＭＯ－１号液组早（Ｐ＜０．０５），说明该实验浓度的川芎嗪不能延长肾保存时间，但有利于移植肾功能的早期恢复。     

温度对WMO-1号液保存犬肾的影响

目的 探讨温度对ＷＭＯ－１号液保存犬肾的影响。方法 采用保存－自体移植模型,比较了不同温度条件下ＷＭＯ－１号液保存犬肾２４小时后的肾功能和形态学变化。结果 保存肾的损害程度与保存温度有关。１５℃保存２４小时不足以防止肾脏缺血损害。５－８℃组术后最大血肌酐值和平均血肌酐值均显著低于１－４℃组和９－１２℃组,形态学变化亦较轻。结论 ＷＭＯ－１号液保存犬肾的最适宜保存温度为５－８℃,保存温度过低或过高均     

不同保存液保存小肠的效果评估

目的 评价乳酸林格（ＬＲ）液、Ｅｕｒｏ－Ｃｏｌｌｉｎｓ（ＥＣ）液、高渗枸橼酸嘌呤（ＨＣ－Ａ）液和武汉医学院器官１号（ＭＷＯ－１）液保存猪小肠的效果。方法 根据不同保存液将动物随机分成４组,每组又根据保存时间不同分成３个亚组,供肠保存２后行自体移植。结果 小肠保存１０小时后移植,术后３周各组受体的存活率均为１００％;保存１８小时,ＬＲ液组、ＥＣ液组、ＨＣ＝Ａ液组和ＷＭＯ－１液组受体的存活率分别为６０     

HX—3液和UW液保存大鼠肝脏效果的比较

采用大鼠肝脏非循环离体灌注模型比较自制的ＨＸ－３液和ＵＷ液对大鼠肝脏的保存效果。实验结果显示，经ＨＸ－３液原位灌洗并保存４８小时的肝脏肝组织含水量正常，而同等条件下换用ＵＷ液，肝组织的含水量虽无明显变化，但都低于正常值；随着保存时间的延长，两组肝窦内皮细胞死亡率逐渐上升，但在２４小时以内两组肝窥内皮细胞死亡率的差异不显著．     

长征-1号多器官保存液低温保存大鼠肺脏并移植的实验研究

目的 通过大鼠原位肺移模型,研究长征－１号多器官保存液保存肺的效果。方法 将大鼠随机分为３组;对组照;以平衡液灌洗供肺,并立即移植;ＣＺ－１液组;以ＣＺ－１液灌洗供肺,保存８小时后移植;Ｅｕｒｏ－Ｃｏｌｌｉｎｓ液组,以Ｅｕｒｏ－Ｃｏｌｌｉｎｓ液灌洗供肺,保存８小时后移植。     

Chemoimmunoprophylaxis of an experimental bladder cancer with retinoids and Bacillus Calmette Guérin

The function of kidneys stored for 48 to 72 hours in hypertonic crystalloid, intracellular solution, Euro-Collins (360 mOsm./l.), was compared with those stored in a new hyperosmolar (520 mOsm./l.) colloid solution designated as TP-II. The TP-II solution contained less K2HPO4 (1.05 gm./l.) and KH2PO4 (317 gm./l.) and more glucose (40 gm./l.) than the Euro-Collins, and also had an albumin concentration of 4.3 gm./dl. Kidneys obtained from beagle dogs were flushed with either Euro-Collins or TP-II solution (4C). Hypothermic storage followed for either 48 or 72 hours, prior to autotransplantation into the iliac fossa and contralateral nephrectomy. Four experimental groups were transplanted as follows: group 1 (n = 8) and group 2 (n = 7) received kidneys that were flushed with 250 ml. of Euro-Collins or TP-II solution, respectively, prior to 48 hours hypothermic storage. Group 3 (n = 5) and group 4 (n = 5) received kidneys that were flushed in the same way as those in groups 1 and 2, respectively, but were stored for a 72-hour period. TP-II appears to be superior to Euro-Collins for hypothermic storage of kidneys for periods as long as 48 hours. When hypothermic storage times are extended to 72 hours, the number of kidneys with normal function after transplantation is reduced for both solutions, however, TP-II solution has a slight advantage over Euro-Collins solution. Further studies will hopefully clarify this issue and lead to the application of TP-II in the clinical setting.     

Influence of acute tubular necrosis on first cadaver kidney transplant function

Prolonged cold storage following intracellular electrolyte flushing increases the probability of significant acute tubular necrosis after cadaver kidney transplantation. The renal function of primary cadaver kidney grafts was compared in 68 recipients who required dialysis and 92 who did not require dialysis during the first week after transplantation. All kidneys were retrieved from beating-heart cadaver donors by our center, flushed with ice-cold intracellular electrolyte solution and cold-stored until transplantation at our hospital. Recipients requiring dialysis during the first week after transplantation received kidneys with a significantly longer cold storage time (27.4 plus or minus 10.2 versus 23.2 plus or minus 7.6 hours) and had significantly higher 1-month serum creatinine nadirs (2.1 plus or minus 1.3 versus 1.5 plus or minus 0.6 mg./dl.). Actuarial kidney graft survivals and serum creatinine levels 1 to 5 years after grafting were not significantly different. Acute tubular necrosis following primary cadaver kidney transplantation does not adversely affect long-term function of kidney grafts flushed with intracellular electrolyte solution and cold-stored until transplantation.     

应用钙离子拮抗剂保护离体肾的实验研究

目的 探讨钙离子拮抗剂对离体肾的保护作用。方法 以3月龄的日本纯种大耳兔为实验对象,分为正常组、实验对照组和实验组,实验对照组将肾脏置于4℃高渗枸橼酸盐腺嘌呤器官保存液（HC－A）中保存24h,实验组将肾脏置于4℃含维拉帕米的HC－A液中保存24h。正常组肾切取即刻测定肾细胞线粒体内钙离子及肾组织中三磷酸腺苷（ATP）的含量;实验对照组和实验组保存24h后测定肾细胞线粒体内钙离子及肾组织中ATP的含量。结果 与肾脏切除后立即检测组相比,实验对照组肾细胞线粒体内Ca∧2＋含量明显上升（P＜0．01）,ATP明显下降（P＜0．01）;实验组上述改变可得以显著改善（P＜0．01）。结论 在离体肾保存中,应用钙离子拮抗剂维拉帕米可阻止Ca∧2＋进入细胞线粒体内,防止能量消耗,从而保护离体肾。     

Cold storage induces time-dependent F2-isoprostane formation in renal tubular cells and rat kidneys

BACKGROUND: Previous findings suggest a possible role for free radicals in cold-storage-associated tissue injury. Because free radical-induced lipid peroxidation catalyzes the cyclooxygenase-independent formation of vasoconstrictive F2-isoprostanes, the hypothesis that isoprostanes are produced during cold storage was tested in this study. METHODS: Total isoprostanes (free and esterified) in renal tubular epithelial (LLC-PK1) cells or whole kidneys, subjected to cold storage, were quantitated employing the gas chromatographic-mass spectroscopic method. LLC-PK1 cells were stored at 4 degrees C in a University of Wisconsin (UW) solution for 0, 24, 48, and 72 hours or 48 hours with desferrioxamine (DFO) or the lazaroid compound 2-methyl aminochroman (2-MAC). In the rat model, kidneys were perfused and stored for 48 hours in the UW solution with or without added DFO or 2-MAC. RESULTS: Isoprostanes in LLC-PK1 cells increased by fivefold following 24 hours of cold storage (36 +/- 2 pg/well to 185 +/- 6, mean +/- SE, following 24 hours of cold storage, P < 0.0001), and the levels continued to increase significantly at 48 and 72 hours. DFO and 2-MAC caused significant suppression of isoprostane formation. Cold storage of the kidneys in UW solution for 48 hours was accompanied by an eightfold increase in isoprostanes compared with control kidneys not subjected to cold storage (25.0 +/- 3.0 vs. 2.9 +/- 0.1 ng/g, P < 0.0001). The addition of 2-MAC or DFO to the UW solution was associated with a near complete suppression of 48-hour cold-induced isoprostane formation. CONCLUSION: Our findings provide evidence for the formation of large quantities of antioxidant-suppressible isoprostanes in kidney cells and whole kidney during cold-preservation. Based on this, it is hypothesized that (a) isoprostanes, which are potent renal vasoconstrictors, may contribute to immediate post-transplant vasoconstriction and dysfunction in kidneys subjected to extended cold storage, and that (b) the addition of 2-MAC or DFO to a UW solution in such circumstances may attenuate these alterations partly by suppressing isoprostane formation.     

自制多器官保存液对草犬肾脏低温保存的实验研究

目的 研究自制上海多器官(SMO)液对草犬肾脏低温保存的效果。方法 采用草犬肾脏非循环离体灌注模型，比较SMO液、Uw液和高渗枸椽酸盐嘌呤溶液(HC-A)对草犬肾脏保存后，其形态学及生物化学的改变，并进行肾脏移植，观察移植后肾功能恢复情况。结果 SMO液保存72h之内，肾脏的形态学无明显改变，其各项生化功能SMO组明显优于HC-A组(P＜0．05)；与Uw液组比较，肾脏线粒体呼吸控制率(RCR)、皮质ATP含量两组间差异有显著性(P＜0．05)；保存48、72h后保存液PH值，SMO组明显高于Uw组(P＜0.05)。结论 SMO液保存犬肾72h效果明显优于HC-A液，与UW液相当。     

Prolonged human kidney preservation by intracellular electrolyte flush followed by cold storage

No kidney transplant center responding to a kidney preservation questionnaire would accept a kidney flushed with an intracellular electrolyte solution and cold-stored for over 40 hours. This study from one center is a comparison of 50 primary cadaver kidney grafts preserved with an intracellular electrolyte flush followed by cold storage for 40 to 61 hours to 82 primary cadaver kidney grafts preserved by the same method for 9 to 24 hours. Kidneys cold-stored for over 40 hours had a significantly increased requirement for dialysis in the 1st week following transplantation (82% versus 34%) and a significantly increased 1-month serum creatinine nadir (2.3 mg/dL versus 1.7 mg/dL). Actuarial graft survivals and serum creatinine levels at 1, 2, and 3 years after grafting were not significantly different. Cadaver donor methylprednisolone (30 to 60 mg/kg) two to nine hours prior to kidney removal significantly reduced the requirement for 1st-week hemodialysis in the kidneys cold-stored for over 40 hours (60% versus 91%). Kidneys preserved by flushing with cold intracellular electrolyte solution can be successfully transplanted after over 40 hours of simple cold storage when the warm ischemia time is very short.     

Evaluation of a preservation solution containing fructose-1,6-diphosphate and mannitol using the isolated perfused rat kidney. Comparison with Euro-Collins and University of Wisconsin solutions

The renal preservation ability of a flushing solution (F-M)with fructose-1,6-diphosphate (1 g/dl) and mannitol (2 g/dl)during cold ischaemia was studied with the isolated perfusedrat kidney model and compared with the Euro-Collins (EC) andUniversity of Wisconsin (UW) solutions. Kidneys were storedin hypothermia for 4 and 18 h after initial flushing with thesolution being tested, and then reperfused at 37°C in anisolated perfusion circuit for 90 min with a Krebs-Henseleitsolution containing 4.5% albumin. Forty-four kidneys were studied and divided in a control groupand six study groups according to the cold ischaemia time andflushing solution used. Renal functional parameters of plasmaflow rate (PFR), renal vascular resistance (RVR), urine flowrate (UFR) glomerular filtration rate (GFR), fractional (FRNa)and net (TNa) sodium reabsortion were assessed during reperfusion.Conventional histology and malon-dialdehyde tissue levels (MDA)were also evaluated. Our results show that PFR, RVR, and UFR were similar in allstudy groups. After 4 and 18 h of cold ischaemia, GFR, FRNaand TNa were better, and conventional histology worse in F-Mthan in EC flushed kidneys. After 4 and 18 h of cold ischaemia,GFR, FRNa and TNa, in fact, were not different between F-M andUW flushed kidneys. After 4 h of cold ischaemia, conventionalhistology was similar in F-M and UW flushed kidneys. Nevertheless,after 18 h of cold ischaemia, UW flushed kidneys showed worsehistological parameters than F-M flushed kidneys. After 4 hof cold ischaemia, MDA was similar in kidneys flushed with thethree solutions. After 18 h of cold ischaemia MDA was higherin EC than in F-M or UW flushed kidneys. In summary, our newly developed cold storage solution showspromising results in renal preservation and its ability to preserveis at least as good as UW solution assessed in the isolatedperfused rat kidney.     

Carolina rinse solution minimizes kidney injury and improves graft function and survival after prolonged cold ischemia

BACKGROUND: Kidney damage caused by cold ischemia-reperfusion injury promotes adverse outcomes after renal transplantation. The purpose of this study was to determine whether Carolina rinse solution (CRS) used at the end of cold ischemic storage decreases kidney injury and improves graft function and survival. METHODS: Inbred male Lewis rats were used as donors and recipients. Left kidneys were removed from donor rats, infused with cold University of Wisconsin solution, and stored for 24, 30, or 48 hr at 0-1 degrees C. Just before implantation, kidneys were flushed with either Ringer's solution or CRS at 35-37 degrees C or were not treated. Kidneys were then transplanted into recipient rats with removal of both native kidneys. RESULTS: Survival and renal function were analyzed over a 14-day postoperative period. Among rats receiving kidneys after 24-hr cold storage, creatinine clearance was 75% greater in rats transplanted with kidneys flushed with CRS compared with Ringer's solution or nontreatment. In animals receiving kidneys after 30-hr cold storage, recipient survival after CRS was significantly higher than with Ringer's solution or without flushing (80% vs. 25% and 17%, respectively). However, CRS failed to prevent renal graft failure after 48 hr of cold storage (14% survival with CRS vs. 0% with Ringer's solution). In separate ex vivo studies, nonviable cell nuclei were labeled by trypan blue after cold preservation and brief warm reperfusion. CRS decreased podocyte and peritubular endothelial cell killing associated with cold ischemia-reperfusion injury. CONCLUSION: Flushing renal explants with warm CRS before implantation diminishes cold ischemia-reperfusion injury and improves the function and survival of transplanted kidneys.