Synergistic effect of donor-specific blood transfusion and a short course of deoxyspergualin in rat kidney transplantation

Deoxyspergualin (DSG), an analogue of spergualin produced by B. laterosporus, has a strong immunosuppressive effect in various transplantation models. We have investigated the mechanism of donor-specific prolongation of survival time in rat kidney grafting by donor-specific blood transfusion (DST) and a short course of DSG. Lewis (LEW) kidney allografts were transplanted into fully allogeneic BN rats. Fresh, whole LEW blood 1.0 ml, was injected i.v. into BN rats 2 days prior to transplantation. Then, DSG, 6 mg/kg per day, was administered by i.m. injection on days 0, 1, and 2 after transplantation. The recipients were divided into five groups: group 1 (n=6) no treatment: group 2 (n=6) DST only; group 3 (n=7) DSG only; group 4 (n=7) DST and DSG; and group 5 (n=6), third party (ACI rats) blood transfusion and DSG. Lymphocytes (cervical lymph nodes) and serum were harvested from BN recipients on day 7 postgrafting. For suppressor cell assays, lymphocytes from BN recipients in each group were added as a third cell to the mixed lymphocyte reaction (MLC) between nontransplanted BN lymphocytes (responder) and LEW or other third party (PVGC, ACI, WKA rats) lymphocytes (stimulator). Antidonor lymphocytotoxic antibody (ADLA) was checked by microcytotoxicity assays. Median survival times (MST) for each group were: group 1, 10 days; group 1, 10 days; group 3, 13 days; group 4, 75 days; and group 5, 13 days. Remarkable prolongation of MST was only noted in group 4. In the suppressor cell assay, group 4 showed significant suppression (40%; P<0.05); the other groups did not show any suppression. This suppressive activity in group 4 was effective only during the MLC between BN and LEW, not during the MLC of third party-BN combinations. Thus, suppressor cells from DST/DSG-treated BN recipients appear to be donor-specific. In the microcytotoxicity assay, the only group that showed any ADLA was group 2, which was not treated with DSG. These results clearly show that both induction of donor-specific suppressor cells and inhibition of ADLA production are associated with the remarkable donor-specific prolongation of kidney allograft survival in DST/DSG-treated recipients.     

Donor-specific renal,but not cardiac,allograft tolerance promotes engraftment of the normally rejected rat skin graft

This study examined whether a heart or kidney graft could provide protection for the more resistant skin graft. Buffalo rat recipients were given a single dose of RIB 5/2 (non-depleting anti-CD4 mAb) plus i.v. Lewis splenocytes 21 days before being given Lewis heart or kidney grafts. Lewis skin was grafted either simultaneously with, or after, long-term (>50 days) Lewis heart or kidney allograft acceptance. Immune responsiveness was analyzed by in vitro mixed lymphocyte culture (MLC), cytotoxic T lymphocytes (CTLs), and limiting dilution analysis (LDA). While i.v. alloantigen plus RIB 5/2 resulted in long-term acceptance of heart and kidney, survival of skin grafts alone was not prolonged. However, simultaneous transplantation with kidney, but not heart, resulted in long-term skin graft acceptance, while skin grafts subsequently grafted to recipients tolerant to kidney or heart were not accepted. In vitro analysis revealed a down-regulation of proliferation, cytotoxicity, and precursor T-helper cells (pThs)/precursor cytotoxic T lymphocytes (pCTLs) in Buffalo recipients accepting Lewis kidney and skin allografts. While RIB 5/2 plus Lewis splenocytes do not prolong the survival of skin grafts, Lewis skin grafted simultaneously with a kidney, but not heart, is accepted indefinitely and provides donor-specific protection for a subsequent skin graft.     

Analysis of suppressor T cells induced by donor-specific transfusion (DST): establishment of a human T cell hybridoma producing an antigen-nonspecific suppressor factor

Formation of suppressor T cells (Ts) induced by donor-specific transfusion (DST) is one of the most commonly suggested mechanisms for the beneficial effect of DST. In this study, we established a human T cell hybridoma derived from the peripheral blood lymphocytes (PBL) of a DST-treated patient, which produced an antigen-nonspecific suppressor factor. Post-DST PBL were fused with an azaguanine-resistant mutant of a human T cell leukemia cell line, CCRF-CEM^AG. After selection and cloning, we established one clone producing the mixed lymphocyte reaction (MLR) inhibitory factor (C524: 18%–43 % suppression). Suppressive activity of the supernatant obtained from C524 after activation by PHA was highly augmented (64%–88 % MLR suppression). This factor inhibited MLR dose-dependently in an antigen-nonspecific and HLA non-restricted manner. These results indicated that Ts clones could be generated in patients receiving DST and that the immunoregulatory factors produced by activated clones may play a role in the prolongation of renal allograft survival.     

Red blood cell transfusion following burn

A severe burn will significantly alter haematologic parameters, and manifest as anaemia, which is commonly found in patients with greater than 10% total body surface area (TBSA) involvement. Maintaining haemoglobin and haematocrit levels with blood transfusion has been the gold standard for the treatment of anaemia for many years. While there is no consensus on when to transfuse, an increasing number of authors have expressed that less blood products should be transfused. Current transfusion protocols use a specific level of haemoglobin or haematocrit, which dictates when to transfuse packed red blood cells (PRBCs). This level is known as the trigger. There is no one 'common trigger' as values range from 6 g dl(-1) to 8 g dl(-1) of haemoglobin. The aim of this study was to analyse the current status of red blood cell (RBC) transfusions in the treatment of burn patients, and address new information regarding burn and blood transfusion management. Analysis of existing transfusion literature confirms that individual burn centres transfuse at a lower trigger than in previous years. The quest for a universal transfusion trigger should be abandoned. All RBC transfusions should be tailored to the patient's blood volume status, acuity of blood loss and ongoing perfusion requirements. We also focus on the prevention of unnecessary transfusion as well as techniques to minimise blood loss, optimise red cell production and determine when transfusion is appropriate.     

骨科手术中自体血液回输对血清前炎性细胞因子浓度的影响

目的 测定下肢骨科手术术野回收血经自体血回收机处理前后及患者自血回输前后血清前炎性细胞因子浓度,观察骨科手术中自体血回输对患者细胞免疫的影响.方法 30例择期行下肢骨科手术患者,分别采集自体血回收机处理前后的术野回收血,并于自体血回输前10min、回输后1 h采集患者动脉血,采用放射酶联免疫吸附测量法测定血样中3种前炎性细胞因子白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)浓度,并观察相关并发症.结果　术野回收血经自体血回收机处理前后3种前炎性细胞因子IL-1β、IL-6、TNF浓度分别为(0.54 ±0.22)、(0.71±0.16)、(16.23±5.68)μg/L和(0.26±0.12)、(0.29±0.09)、(6.32±2.57)μg/L,与处理前比较,处理后3种细胞因子浓度显著降低(P＜0.05);自血回输前后患者血清中3种细胞因子IL-1β、IL-6、TNF浓度分别为(0.35±0.17)、(0.47±0.15)、(8.44±3.56)μg/L和(0.39±0.19)、(0.52±0.18)、(9.48±3.45)μg/L,与回输前比较,回输后患者血清中3种细胞因子浓度增高(P＜0.05);30例患者自体血回输后12 h内均未观察到低血压、心动过速、血红蛋白尿、凝血功能紊乱、脓毒血症、空气栓塞、心肺问题等并发症.结论　骨科手术患者术中可适量自体血回输,回收血液经自体血回收机处理后前炎性细胞因子浓度显著降低,回输后未观察到严重并发症.

Abstract：

Objective To investigate the effects of autologous blood transfusion on serum cytokine levels in patients undergoing lower limb orthopedic surgery. Methods A total of 30 cases scheduled for undergoing lower limb orthopedic surgery were enrolled in this study. Each patient had four blood samples taken (pre-transfusion, one h post-transfusion, cell saver container, and post-filtration from the blood bag). An enzyme linked immunosorbent assay (ELISA) measurement of radiation was conducted to determine levels of the cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF). Serious complications and sequelae associated with autotransfusion were recorded. Results In comparison to cell saver container, levels of IL-1β, IL-6 and TNF in the blood bag were decreased significantly (P＜0. 05 ). In comparison to pre-transfusion, levels of IL-1β, IL-6 and TNF were increased significantly (P ＜ 0. 05 ). No serious complications and sequelae associated with autotransfusion were observed. Conclusion The use of cell saver container appears to be safe in patients undergoing orthopedic surgery and the levels of the cytokines in post-filtration blood are decreased.     

回收式自体输血与异体输血对机体炎症反应的影响

背景 自体血回输在围术期应用日益广泛.自体血和异体血输注均会对机体的炎症反应产生一定的影响.目的 对两种输血方式后机体炎症因子的变化进行对比,阐述围术期两种输血方式对患者的影响. 内容 异体血在输注过程中产生大量炎症因子,如白细胞介素(interleukin,IL)-2、IL-6、IL-8、IL-10、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)等,引起机体炎症反应,可能是术后并发症的重要原因.自体血在回收、过滤的过程中同样会产生一定量的炎症因子. 趋向 对骨科自体血及异体血应用进行对比,研究两种输血方式对机体炎症反应的影响.     

Role of MLC serum inhibitory factors in high MLC-reactive kidney transplant recipients pretreated with donor-specific blood transfusion (DST)

In order to clarify the beneficial effect of donor-specific blood transfusions (DST) on kidney allograft survival, sera from 16 patients treated with DST were studied using the mixed lymphocyte culture (MLC) serum inhibition test. The results demonstrate that MLC inhibitory factors could be induced in the serum of the recipients after the completion of DST, and that these factors are directed against cells of the recipient but not against cell from the donor. Regarding the correlation with rejection episodes and clinical outcome, a significant improvement in renal transplant survival and reduction in rejection episodes was observed when MLC inhibitory factors were present in post-DST sera. These data suggest that such factors may contain antibodies directed against recognition sites on T lymphocytes, e.g., antiidiotypic antibodies, and the associated with prolonged graft survival of living-related, high MLC-reactive one-haplotype-mismatched kidney.     

大量输血患者成分输血分析

目的探讨成分输血在大量输血患者中的合理应用。方法选择笔者所在医院2009~2010年60例大量输血患者,分析红细胞悬液、血浆、血小板、冷沉淀等各种成分血的使用比例。结果 60例大量输血患者中以输注红细胞悬液最多,占33.7%;血浆占22.4%。结论大量输血患者在各种成分血的用量上有很大差异,应根据患者的出血情况和实验室检查结果输注不同的血液制品。     

Intraoperative autotransfusion is associated with modest reduction of allogeneic transfusion in prosthetic hip surgery

Background: The efficacy of intraoperative salvage and washing of wound blood and the predictors of allogeneic red cell transfusions in prosthetic hip surgery are insufficiently known.
Methods: In 96 patients, undergoing primary or revision surgery, salvaged and washed red cells and, if necessary, allogeneic blood were used to keep haematocrit not lower than 33%. The bleeding of red cells during hospital stay was calculated from the red cell balance. The preoperative red cell reserve (millilitres of red cells in excess of a haematocrit of 33%) was estimated and the difference between this volume and the total bleeding of red cells was retrospectively used to classify patients with regard to the need for red cells. Stepwise regression analysis was used to define patient-related variables associated with allogeneic blood transfusion.
Results: Preoperative knowledge of the type of operation (primary, revision), the preoperative red cell reserve, and the body mass could predict roughly half of the need for banked blood (r²=0.45). Only one-third of the total bleeding of red cells was retransfused. For complete avoidance of allogeneic blood, autotransfusion was most effective in patients with a moderate need (0–4 u). However, 32% of such patients required allogeneic blood.
Conclusions: Autotransfusion has a limited efficacy to decrease the need for allogeneic blood, and other blood-saving methods should be added for this purpose. It is difficult to predict the need for allogeneic blood preoperatively.     

大鼠小体积肝脏移植后早期肝内细胞因子的变化

目的 研究不同冷缺血条件下大鼠小体积肝移植(30%标准体积)后早期肿瘤坏死因子α(TNF-α)及白细胞介素6(IL-6)的变化,及其与肝脏再生的关系.方法 建立Lewis大鼠30%标准体积的原位肝移植模型.根据供肝在UW液中冷保存时间的不同,将受者分为3组:冷缺血1 h组、冷缺血8 h组和冷缺血16 h组,每组均为20只.观察受者存活情况至术后第7天,并分别在移植肝恢复血流后90 min、1 h、2 h、4 h和7 d收集样本,检测移植肝组织中TNF-α和IL-6表达情况,肝细胞DNA的合成情况,进行移植肝的形态学观察.结果 大鼠肝移植手术成功率均为100%.移植后第7天,冷缺血1 h和8 h组受鼠的存活率均为100%.冷缺血16 h组受鼠的存活率较低,移植后第7天无受鼠存活.冷缺血1 h组TNF-α和IL-6的表达水平较低,冷缺血8 h组和冷缺血16 h组TNF-α和IL-6的表达则高于冷缺血1 h组(F=58.81和F=184.12,P<0.05).冷缺血8 h组和冷缺血16 h组间TNF-α和IL-6的表达的差异无统计学意义.冷缺血1 h组增殖细胞数目明显高于冷缺血8 h组,差异有统计学意义(t=5.59,P<0.05).移植术后24h,冷缺血1 h组移植肝有轻度的组织学损伤;冷缺血8 h组移植肝有轻度的窦状隙扩张和轻度的炎症;冷缺血16 h组移植肝有局部淤血,存在肝细胞崩解和坏死等改变.结论 在小体积肝移植后早期,TNF-α和IL-6的上调表达对肝脏再生有重要意义.不同冷缺血时间的小体积肝脏移植物内存在早期启动肝脏再生的信号.     

Anaphylactoid reaction associated with blood transfusion during anesthesia

Key words  anaphylactoid reaction - blood transfusion - hemodynamics     

The effect of donor-specific transfusion and cyclosporin A on small bowel transplantation in the rat

Pre-transplant blood transfusions are given as a means of desensitization to reduce the required dose of cyclosporin A (CsA). In this study, the effect of pretransplant blood transfusion on host survival and T-cell function against alloantigen were investigated. Male Lewis rats (RT1¹) were used as the recipients in all experiments, and male DA rats (RT1^a) were used as the blood and small bowel donors, and as a source of allogeneic stimulator cells. Male BUF rats (RT1^b) were used as donors of third party blood, and of allo-stimulator cells in a delayed-type hypersensitivity (DTH) response. In our experimental design, Lewis rats were divided into the following groups according to the type of administration: (1) a donor-specific blood transfusion (DST) 8 days preoperatively and a concurrent 5-day course of CsA at 10 mg/kg per day; (2) a nonspecific third party blood transfusion (NST) and CsA at 10 mg/kg per day from day 8 to day 4 preoperatively; (3) CsA alone from day 8 to day 4 preoperatively; (4) DST alone 8 days preoperatively; or (5) no treatment, being the control group. Postoperative treatment consisted of CsA at 2.5 mg/kg per day for 30 days. Rats conditioned with NST plus CsA, CsA alone, DST alone, and the untreated control rats survived for 7.2 ±1.2, 9.0 ± 2.2, 6.8 ± 0.4, and 7.4 ± 1.6 days, respectively. In contrast, the five rats conditioned with DST plus CsA survived for 100 days or more. This study demonstrates that long-term survival of a small bowel allograft can be achieved by host-conditioning with a combined treatment of DST and low-dose CsA.This paper was presented at the 10th Congress of the Asian Association of Pediatric Surgeons held in Seoul, Korea from March 26–29, 1990, and at the 90th Annual Meeting of the Japanese Surgical Society held in Sapporo, Japan in 1990.     

急诊手术围手术期输血与术后感染及并发症的关系

目的 分析急诊手术患者围手术期输血与术后感染的关系。方法 收集2011年至2015年部分急诊手术住院患者病历资料,包括患者基线资料、围手术期临床和实验室数据以及住院期间并发症资料。输血的定义范围为从患者入院到出院的任何输血事件。主要记录术后感染性事件,包括切口和手术部位感染、伤口裂开、尿路感染、肺炎、败血症和感染性休克;次要记录包括住院时间、非计划气管插管、呼吸机使用超过48小时、急性肾衰竭,任何血栓栓塞事件和意外再次手术探查等。结果 在收集到的3153例急诊手术患者中,共242例（7.7%）接受输血治疗,接受输血患者年龄大于无输血患者,输血组BMI低于未输血组;输血患者ASA分类3级和4级低于未输血组;输血患者的平均总手术时间、总住院时间比无输血组长;输血患者发生血栓栓塞并发症的风险,如肺栓塞、深静脉血栓、呼吸功能障碍的风险增高;输血患者更有可能进行非计划气管插管。本组病例共594例患者（18.83%, 594/3135）发生术后感染事件,其中输血患者的感染率占所有输血者的39.7%（96/242）;糖尿病、COPD、慢性心脏病和高血压等慢性疾病史有显著性差异;体重减轻超过10%、较高ASA评分及污染严重伤口更容易感染;皮质类固醇使用、出血性疾病的也更容易感染。此外,低蛋白血症、低血细胞比容和也存在显著差异;开放手术感染率高于腹腔镜手术。结论 急诊手术患者接受输血与术后感染性并发症的风险增加有关。     

诱导型一氧化氮合酶与Th1/Th2型细胞因子在不明原因早期自然流产绒毛和蜕膜组织中的表达

目的了解不明原因早期自然流产局部绒毛、蜕膜组织中诱导型一氧化氮合酶(iNOS)和辅助性T细胞(Th1)/Th2型细胞因子的表达,进一步探讨不明原因早期自然流产发病机制。方法Th1/Th2型细胞因子分别以白细胞介素(IL)-2和IL-10为代表,采用原位杂交及免疫组化方法对40例自然流产患者绒毛和蜕膜中iNOS、IL-2、IL-10的表达(试验组)进行检测,利用计算机CMIAS系统,表达指标以阳性数密度(N/S)计算,并与40例正常人工流产的绒毛和蜕膜组织(对照组)比较。结果试验组iNOS、IL-2、IL-10原位杂交显示绒毛、蜕膜组织分别为(0.082±0.026)、(0.0026±0.0007)、(0.036±0.011)vs(0.10±0.03)、0、(0.02±0.006),对照组分别为(0.022±0.007)、(0.0028±0.0007)、(0.042±0.011)vs(0.03±0.009)、0、(0.032±0.006);两组比较iNOS、IL-10差异有显著意义(P<0.05)。免疫组化结果显示与原位杂交结果一致。结论INOS和ThI/Th2型细胞因子与自然流产相关,可能是介导流产发生的重要因素之一。