CTLA4-Ig和抗CD40L单克隆抗体抑制大鼠胰腺移植急性排斥反应的机制

目的探讨细胞毒T淋巴细胞相关抗原4免疫球蛋白（CTLA4-Ig）和抗CD40L单克隆抗体对异基因大鼠胰腺移植后急性排斥反应的作用及其相关机制。方法建立大鼠的胰十二指肠移植模型，供者为F344大鼠，受者为经链尿佐菌素诱导为糖尿病模型的Lewis大鼠，受者移植后分为4组，每组12只。A组：为应用生理盐水对照组；B组：应用CTLA4-Ig200μg；C组：应用抗CD40L单克隆抗体200μg；D组：联合应用CTLA4-Ig和抗CD40L单克隆抗体各200μg。各组分别于术后第2d腹腔注射相应的药物。术后1、4、7、10d分别取各组的移植胰腺，进行常规病理检测；采用逆转录聚合酶链（RT-PCR）法检测移植物白细胞介素2（IL-2）、白细胞介素4（IL-4）、白细胞介素10（IL-10）、γ干扰素（IFN-γ）的表达；术后第1、4、7、10d取受者外周血，采用流式细胞术计数T细胞亚群CD3^＋、CD4^＋和CD8^＋；术后第4d取移植胰计数CD4^＋CD25^＋T细胞。结果病理检测显示：与A组相比，B、C组排斥反应明显减弱，D组几乎未发生排斥反应；B、C、D组IL-2的表达高峰延迟，且表达水平较A组有不同程度的降低，D组又较B、C组表达水平下降，差异有统计学意义；B、C、D组IFN-γ的表达水平较A组有不同程度的降低，但D组与B、C组的差异不显著；B、C组IL-4的表达水平较A组有不同程度的升高，D组较A、B、C组表达水平下降，差异有统计学意义；B、C组IL-10的表达水平较A组有不同程度的升高，D组与A组差异不显著；B、C、D组CD3^＋、CD4^＋和CD8^＋T细胞数均相对减少，CD4^＋CD25^＋T细胞数有不同程度升高，与A组比较，D组的差异最为显著。结论联合应用CTLA4-Ig和抗CD40L单克隆抗体能更有效地抑制大鼠胰腺移植后排斥反应，其机制可能与Th1／Th2型细胞因子偏移及CD4^＋CD25^＋调节性T细胞增多等有关。     

联合应用CTLA-4Ig及抗ICOS单克隆抗体延长小鼠移植胰岛存活时间

目的探讨共刺激信号阻断剂细胞毒T淋巴细胞相关抗原4免疫球蛋白（CTLA-4Ig）及抗共同刺激分子ICOS单克隆抗体（ICOSmAb）对移植胰岛功能的影响。方法以BALB／c小鼠为供者，C57BL／6糖尿病小鼠为受者，进行同种胰岛细胞移植。将移植后的小鼠随机分成4组，每组10只。ICOS组：移植后1、3、5d腹腔内注射ICOSmAb 100μg／kg；CTLA4组：移植后0、2、4d腹腔内注射CTLA-4Ig50μg／kg；联合阻断组：移植后腹腔注射CTLA-4Ig和ICOSmAb，用法同CTLA4组和ICOS组；对照组：单纯胰岛移植，不注射CTLA-4Ig和ICOSmAb。观察术后移植物存活时间和移植胰岛的病理改变；逆转录聚合酶链法（RT-PCR）检测移植胰岛组织中白细胞介素2（IL-2）、白细胞介素10（IL-10）mRNA的表达情况；应用流式细胞仪检测CD4^＋、CD8^＋T淋巴细胞表达情况。结果联合阻断组的小鼠移植胰岛存活时间较其它3组明显延长，移植胰岛的细胞形态经光镜检查接近正常。联合阻断组与其它3组比较，IL-2mRNA表达减少，差异有统计学意义（P〈0．05），IL-10mRNA的表达差异无统计学意义（P〉0．05）；移植术后21d，CD4^＋、CD8^＋T淋巴细胞表达上调不明显。结论应用CTLA-4Ig和ICOSmAb联合阻断CD28和共同刺激分子ICOS，可以明显的抑制排斥反应，延长移植胰岛的存活时间及存活率。     

联合阻断CD28和可诱导共刺激分子对小鼠同种异体胰岛移植物急性排斥反应的影响

目的 在小鼠同种异体胰岛移植模型上，通过联合阻断CD28和可诱导共刺激分子（ICOS），观察CTLA-4Ig及ICOS单克隆抗体（ICOSmAb）对胰岛移植物功能的影响。方法将小鼠随机分成4组，每组10只。联合阻断组：分别于移植后0．2、4d和1、3、5d腹腔内注射CTLA-4Ig50μg／kg和ICOSmAb100μg／kg体重组。ICOS组：移植后1、3．5d腹腔内注射ICOSmAb100μg/kg体重组。CTLA4组：移植后0、2、4d腹腔内注射CTLA-4Ig50μg/kg体重组。对照组：单纯胰岛移植，不做CT-LA-4Ig和ICOSmAb处理组。观察术后移植物存活时间和病理改变，RT-PCR检测移植组织中IL-2、IL-10mRNA表达，流式细胞仪检测CD4^＋、CD8^＋T淋巴细胞表达。结果 联合阻断组小鼠移植物存活时间平均为（31．00±6．57）d，较其他3组延长，差异有统计学意义（P〈0．05）；IL-2和IL-10mRNA表达水平分别为（44．23±6．24）和（65．23±11．02），与其他3组比较，IL-2mRNA表达差异有统计学意义（P〈0．05），而IL-10mRNA表达差异无统计学意义（P〉0．05）；CD4^＋、CD8^＋T淋巴细胞荧光阳性百分率分别为（13．73±0．49）％和（14．56±0．31）％，表达上调均不明显；移植胰岛光镜检查接近正常。结论 应用CTLA-4Ig和ICOSmAb联合阻断CD28和ICOS，可以明显抑制胰岛移植物排斥反应。     

PD-L1.Ig与1,25(OH)2D3联合应用对异基因大鼠胰腺移植后急性排斥反应的影响

目的 探讨PD-L1.Ig联合1,25(OH)2D3对大鼠胰腺移植急性排斥反应的免疫调控作用以及对大鼠移植胰腺存活时间的影响.方法 以F344大鼠为供体,Lewis大鼠为受体,建立4组原位胰腺移植模型.每组12例,共48例.A组:对照组、B组:PD-L1.Ig组、C组:1,25(OH)2D3组、D组:PD-L1.Ig+1,25(OH)2D3组;观察术后各组血糖变化,移植物存活以及组织病理学改变;流式细胞检测受体血、脾以及移植胰CD4+T细胞、CD8+T细胞、CD4+CD25+T细胞表达水平;酶联免疫吸附试验(ELISA)检测移植物局部白细胞介素(IL)-2、-4、-10、-12表达水平;于术后第7天分别杀死各组受体和供体鼠各2只,取受体脾细胞与供体作混合淋巴细胞反应(MLR).结果 与A比较,PD-L1.Ig组并未显著延长移植胰腺存活时间(P＞0.05),C、D组对改善血糖水平和延长移植胰腺存活时间作用明显(P＜0.01),其中,D组移植胰腺存活时间最长(23.9±0.8)d;与A组比较,B组无明显改善,C组排斥反应较A组明显减弱,而D组几乎未发生排斥反应;CD3+CD8+T细胞计数D、C、B与A组比较均有减少,差异有统计学意义(P＜0.01).CD4+T细胞D、C、B与A组的差异有统计学意义(P＜0.05).CD4+CD25+调节性T细胞A、B、C、D组逐渐升高,差异有统计学意义(P＜0.01),且以D组较为明显(C组比A组P＜0.01);D组明显抑制移植物局部Th1型细胞因子IL-2、IL-12的产生,显著提高Th2型细胞因子IL-4、10的水平;与对照组比较,各治疗组受体T淋巴细胞在MLR中表现对供体淋巴细胞特异性低反应性,能有效抑制T细胞对同种异体抗原的反应.结论 共刺激阻断剂PD-L1.Ig联合1,25(OH)2D3可有效抑制细胞免疫应答,干预急性排斥反应,显著延长大鼠移植胰腺存活时间.     

肾移植患者外周血CD4^＋CD25^＋调节性T细胞的变化及临床意义

目的观察肾移植患者外周血中CD4^＋CD25^＋调节性T细胞水平的变化，探讨其在诊断移植肾急性排斥反应中的作用。方法采用流式细胞仪检测26例肾移植患者及30例正常对照组外周血中CD4^＋CD25^＋调节性T细胞水平。结果①慢性肾衰竭患者外周血CD4^＋CD25^＋调节性T细胞水平与对照组比较，差异有统计学意义（P〈0．05）。②非排斥组移植后1、2、4、8周CD4^＋CD25^＋调节性T细胞水平明显高于移植前（P〈0．01）。③急性排斥组排斥反应主要发生在术后第7～21d，其CD4^＋CD25^＋调节性T细胞水平明显低于同期的非排斥组（P〈0．01）。结论CD4^＋CD25^＋调节性T细胞水平的测定可以作为肾移植患者移植后急性排斥反应诊断和预测预后的重要指标。     

移植物中CD4+CD25+T细胞的表达与异基因造血干细胞移植后移植物抗宿主病的相关性

异基因造血干细胞移植是治疗恶性血液病的有效手段之一，而其并发症移植物抗宿主病（GVHD）严重影响患者的预后和生存质量。近年来已有研究表明，去除移植物中的CD4^＋CD25^＋T细胞，移植后aGVHD的发生率上升。这说明移植物中CD4^＋CD25^＋T细胞表达水平与GVHD的发生相关。因此，我们对移植物中CD4^＋CD25^＋T细胞亚群比例与受者GVHD的发生进行了初步研究。报告如下。     

移植抗原特异性转基因CD8+T细胞对白细胞介素2的依赖性

目的研究白细胞介素2（IL-2）在调节移植抗原特异性转基因CD8^＋T细胞介导的免疫排斥反应中的作用。方法将经荧光染科CFSE标记后的C57BL／6小鼠和2CTg小鼠（CD4敲除鼠）淋巴细胞分别植入经致死剂量γ射线照射过的两组DBA／2J小鼠体内，检测CD4^＋与CD8^＋T细胞在体内分裂增殖的时相，并用胞浆内IL-2标志染色方法测定活化后T细胞表达IL-2的能力。以Balb／c小鼠为供者，糖尿病2CTg小鼠和2C Tg-IL-2KO小鼠（IL-2敲除鼠）为受者，进行胰岛细胞移植。观察CD8^＋ T细胞在介导移植排斥中的作用。结果DBA／2J小鼠输注了C57BL／6小鼠的淋巴细胞后，CD4^＋与CD8^＋T细胞分裂增殖均非常明显，前者表达大量IL-2，后者则不表达。DBA／2J小鼠输注了2CTg小鼠的淋巴细胞后。在完全没有CD4^＋T细胞存在的情况下，CD8^＋T细胞仍明显分裂增殖并大量表达IL-2。2CTg和2CTg—IL-2KO小鼠移植胰岛细胞后，前者迅速发生排斥反应，胰岛移植物的平均存活时间仅为8d，而后者胰岛移植物的平均存活时间〉50d。结论CD8^＋T细胞在产生和利用IL-2时有很大的可塑性。CD4^＋T细胞存在时，CD8^＋T细胞能有效利用CD4^＋T细来源的IL-2进行分裂增殖，在缺乏CD4^＋T细胞时，则利用自身来源的IL-2进行分裂增殖；移植抗原特异性CD8^＋T细胞的效应功能完全依赖于IL-2，排斥反应由CD8^＋T细胞介导时，阻断IL-2／IL-2受体通路可诱导移植物长期存活。     

细胞毒T淋巴细胞A4Ig抑制大鼠胰腺移植急性排斥反应的研究

B7／CTLA4-CD28是其中两大经典途径之一，我们试图利用细胞毒T淋巴细胞（CTL）A4Ig竞争性阻断B7／CD28通路，探讨其对大鼠胰腺移植急性排斥反应的作用及机制。     

骨髓输注联合阻断共刺激通路延长大鼠皮肤移植物的存活时间

目的 探讨骨髓输注联合阻断共刺激通路对大鼠移植皮肤存活的影响及可能机制.方法 以Lewis大鼠为受者,皮肤移植前经尾静脉输注供者(BN大鼠)骨髓细胞2×108 个,并于骨髓输注当天、输注后第2、4、6及8天腹腔注射抗CD25单克隆抗体,同时按照分组要求腹腔注射细胞毒性T淋巴细胞相关抗原4融合蛋白(CTLA4Ig组)、抗CD154单克隆抗体(抗CD154单抗组)以及CTLA4Ig和抗CD154单克隆抗体(联合处理组),于骨髓输注后第8天移植BN大鼠的皮肤.另以仅行皮肤移植者为对照(对照组).观察各组移植物抗宿主病(GVHD)的发生情况、外周血中供者细胞嵌合率、T淋巴细胞凋亡率及移植皮肤的存活时间.结果 各组均未观察到GVHD的发生.骨髓输注后第7天即可在CTLA4Ig组、抗CD154单抗组及联合处理组观察到嵌合现象,至第21天时,嵌合率仍维持于一定水平,联合处理组明显高于其它三组(P＜0.01).骨髓输注后第7及21天,CTLA4Ig组、抗CD154单抗组及联合处理组间两两比较,T淋巴细胞凋亡率的差异无统计学意义,但均显著高于对照组(P＜0.05,P＜0.01).CTLA4Ig组、抗CD154单抗组及联合处理组移植皮肤的存活时间显著长于对照组(P＜0.01),联合处理组移植皮肤存活时间为(16.7±3.1)d,明显长于CTLA4Ig组和抗CD154单抗组(P＜0.05).结论 骨髓输注联合阻断共刺激通路能够延长移植皮肤存活时间,其机理可能与诱导嵌合和T淋巴细胞凋亡有关.     

CD4-CD8-双阴性T细胞在移植免疫耐受形成中的作用

体内移植物抗宿主反应和宿主抗移植物反应主要是由T淋巴细胞介导。在同种异体移植时,因移植物的组织相容性抗原与受者不符,而刺激受者免疫系统发生排斥反应。由于移植物的细胞构成不同,触发的免疫反应会有所差异。去除T细胞或抑制T细胞的功能均能使移植物的存活时间延长。最近研究发现一种T细胞亚群——αβTCR^＋CD3^＋CD4^- CD8^-双阴性T细胞（double-nega-tiveTcell,DNT）可通过抑制同系基因型CD4^＋ T细胞或CD8^＋ T细胞从而抑制移植物排斥反应的发生,因此研究DNT细胞在移植免疫耐受形成中的作用将有重要意义。     

Bradycardia with sevoflurane in siblings with Branchio-oto-renal syndrome

Branchio-oto-renal syndrome (BOR, Melnick-Fraser syndrome, MIM#113650) refers to a rare autosomal dominant disorder characterized by branchial cysts or fistulas, hearing loss, external ear malformation, preauricular pits and renal abnormalities. The authors present three episodes of significant bradycardia in two siblings diagnosed with BOR syndrome during the sevoflurane general anesthesia. There is no published experience of anesthesia with this syndrome. Bradycardia occurred variously at induction, maintenance and immediately prior to emergence and required surgical stimulation, atropine, or epinephrine to treat. We seek to raise awareness of the potential for bradycardia during the procedures in patients with this syndrome requiring volatile anesthesia, especially sevoflurane.     

Increases in BMD correlate with improvements in bone microarchitecture with teriparatide treatment in postmenopausal women with osteoporosis.

Increases in BMD are correlated with improvements in 2D and 3D trabecular microarchitecture indices with teriparatide treatment. Therefore, improvements in trabecular bone microarchitecture may be one of the mechanisms to explain how BMD increases improve bone strength during teriparatide treatment. INTRODUCTION: Bone strength is determined by BMD and other elements of bone quality, including bone microarchitecture. Teriparatide treatment increases BMD and improves both cortical and trabecular bone microarchitecture. Increases in lumbar spine (LS) BMD account for approximately 30-41% of the vertebral fracture risk reduction with teriparatide treatment. The relationship between increases in BMD and improvements in cortical and trabecular microarchitecture has not yet been studied. MATERIALS AND METHODS: The relationship between increases in BMD and improvements in cortical and trabecular microarchitecture after teriparatide treatment was assessed using data from a subset of patients who had areal BMD measurements and structural parameters from transiliac bone biopsies in the Fracture Prevention Trial. 2D histomorphometric and 3D microCT parameters were measured at baseline and 12 (n = 21) or 22 (n = 36) mo. LS BMD was assessed at baseline and 12 and 18 mo, and femoral neck (FN) BMD was measured at baseline and 12 mo. Pearson correlation was performed to assess the relationship between actual changes in BMD and actual changes in microarchitectural parameters. RESULTS: Changes in LS BMD at 12 mo were significantly correlated with improvements in trabecular bone structure at 22 mo: 2D bone volume (r = 0.45, p = 0.02), 2D mean wall thickness (r = 0.41, p = 0.03), 3D bone volume (r = 0.48, p = 0.006), 3D trabecular thickness (r = 0.44, p = 0.01), 3D trabecular separation (r = -0.37, p = 0.04), 3D structural model index (r = -0.54, p = 0.001), and 3D connectivity density (r = 0.41, p = 0.02). Changes in LS BMD at 18 mo had similar correlations with improvements in bone structure at 22 mo. Changes in FN BMD at 12 mo were significantly correlated with changes in 2D mean wall thickness (r = 0.56, p = 0.002), 3D bone volume (r = 0.51, p = 0.004), 3D trabecular thickness (r = 0.44, p = 0.01), 3D trabecular separation (r = -0.46, p = 0.01), and 3D structural model index (r = -0.55, p = 0.001). CONCLUSIONS: Increases in BMD are correlated with improvements in trabecular microarchitecture in iliac crest of patients with teriparatide treatment. Therefore, improvements in trabecular bone microarchitecture may be one of the mechanisms to explain how BMD increases improve bone strength during teriparatide treatment.     

Pyloroplasty in association with Nissen fundoplication in children with neurologic disorders

Previous studies have suggested that delayed gastric emptying occurs in severely mentally retarded patients with gastroesophageal reflux. Based on this data, pyloroplasty was employed in such patients. A retrospective analysis of 99 consecutive patients who underwent primary fundoplication for GER was performed. Gastric emptying, as measured by successful removal of the nasogastric tube or elevation of the gastrostomy tube, was studied. Children with neurologic disorders had no clinically significant difference in gastric emptying after fundoplication (3.31 days) when compared with neurologically normal patients (2.21 days). When added to Nissen fundoplication, pyloroplasty did not hasten the return of gastrointestinal function in the severely impaired patients (4.91 days). A prospective study employing gastric isotope bolus feedings before and after Nissen fundoplication will determine if pyloroplasty improves gastric emptying when used in conjunction with Nissen fundoplication for patients with severe neurologic disorders.     

Chemotherapy in patients with teratoma with malignant transformation

OBJECTIVE: Germ-cell tumours (GCTs) with a non-GCT malignant component are a unique and rare phenomenon called teratoma with malignant transformation (TMT). The only published series of patients with TMT treated with chemotherapy comprised 10 patients. We report here our experience in treating 14 patients with TMT. PATIENTS AND METHODS: Sarcoma was identified in 10 of 14 patients, with rhabdomyosarcoma ranking first (n=4). Other histological types included adenocarcinoma (n=3) and bronchoalveolar carcinoma (n=1). Immunohistochemistry was performed to help in identifying the malignant non-GCT component. RESULTS: Primary treatment consisted of surgery alone in 4 patients. The remaining 10 patients received first-line cisplatin-based chemotherapy with resection of residual masses (n=5): 4 patients had a complete response and 5 had a partial response. Overall, 9 patients developed a relapse with a median time of 84 mo (range: 6-168). At relapse, 8 patients received a chemotherapy regimen directed to the non-GCT component. Four of these patients achieved a partial response. With a median follow-up of 59 mo (range: 3-180), 4 of 14 patients are alive, including 3 who are disease-free. CONCLUSION: To our knowledge, this is by far the largest reported European series of chemotherapy in TMT. Although TMT has a poor prognosis compared to GCT, its management may be improved by adapted chemotherapy associated with surgical resection of residual masses.     

Bradycardia with sevoflurane induction in patients with trisomy 21

The authors present a series of three paediatric patients with trisomy 21 who developed significant bradycardia during inhalation induction with sevoflurane. All three were undergoing adenotonsillectomy. The possible association of such problems with trisomy 21 and treatment options are reviewed. Although the occurrence of bradycardia in these three patients may have been a random occurrence, given its occurrence in three of five consecutive patients with trisomy 21, a causal relationship may be involved and future observation of these patients appears warranted.     

Atherosclerosis in patients with analgesic nephropathy treated with haemodialysis

SUMMARY: Accelerated atherosclerosis was reported to be associated with chronic analgesic consumption, but most studies were retrospective, and individual findings have almost never been controlled with regard to other atherosclerotic risk factors. Ten haemodialysis patients with analgesic nephropathy (group I) and 19 haemodialysis patients where renal failure was not caused by analgesic nephropathy (group II) were included in the study. All patients were female without diabetes. Using B-mode ultrasonography, we compared intima-media thickness (IMT) in the carotid arteries and plaque occurrence, and their thickness in group I with that in group II. the possible differences in atherosclerotic risk factors in both groups were also investigated. In group I, the average age was 60.2 years, and the average dialysis treatment was 55.7 months. In group II, the average age was 54.6 years, and the average duration of dialysis treatment was 50.4 months. We found no statistically significant difference in the age and duration of dialysis treatment between groups I and II. the IMT values of the carotid arteries (0.97 vs 0.78 mm; P = 0.027) were significantly higher in group I. More patients had plaques in group I (90 vs 57.9%), and the number of plaques ( P = 0.037) and their thickness ( P = 0.043) were significantly higher in this group. There was no statistically significant difference in the atherosclerotic risk factors between groups I and II. the results indicate that patients with analgesic nephropathy treated with haemodialysis showed advanced atherosclerosis compared with other haemodialysis patients, despite no difference being found in the atherosclerotic risk factors between these patients.