血清sPTA1与人活体小肠移植急性排斥反应的相关性评价

目的 观察我国首例父子间活体小肠部分移植术患者围手术期血清可溶性血小板／Ｔ细胞活化抗原１（ｓＰＴＡ１）水平与排斥反应的相关性。方法 用夹心ＥＬＩＳＡ法测定轿清ｓＰＡ１水平。结果 于术后第６７ｄ出现移植物排斥反应。从发生排斥反应前２周开始,ｓＰＴＡ１呈逐渐升高趋势,于排斥反应前５ｄ已达６０．４９ｎｇ／ｍｌ,前１ｄ达７１．６４ｎｇ／ｍｌ。经加强免疫抑制后,血清ＰＴＡ１水平于次日迅速下降。结论 活体部分     

尿趋化因子IP-10和Fractalkine在移植肾急性排斥反应早期诊断中的应用

背景：相关研究已证实,趋化因子IP-10、Fractalkine在器官移植后急性排斥反应过程中发挥着重要的作用。 目的：检测尿液中IP-10和Fractalkine水平变化,并结合肾组织活检病理,探讨尿液中IP-10 和Fractalkine在移植肾急性排斥反应早期诊断中的意义。 方法：106例同种异体肾移植患者,根据移植后临床表现、实验室检查及肾穿刺组织病理学检查结果,分为急性排斥反应组(n=16)和非急性排斥反应组(n=90);另选择健康志愿者作为正常对照组。用双抗体夹心酶联免疫吸附试验检测尿IP-10和Fractalkine浓度变化。 结果与结论：急性和非急性排斥反应组患者在移植后的尿IP-10及Fractalkine的表达水平均较移植前明显升高,但非急性排斥反应组在移植后7 d呈下降趋势,至第11天降至移植前水平,而急性排斥反应组则持续高表达,IP-10在移植后第1天和Fractalkine在移植后第3天即与急性排斥反应组比较,差异有显著性意义(P < 0.05)。提示,肾移植后尿液中IP-10和Fractalkine水平的检测对于急性排斥反应发生的早期诊断和早期治疗具有重要意义。     

肾移植患者尿液MCP-1检测的意义

目的:检测肾移植受者尿液中单核细胞趋化性肽-1(MCP-1)的含量,了解其变化规律,探讨其在肾移植急性排斥反应中的意义。方法:用ABC-ELISA法,检测尿液中MCP-1的含量。结果:肾功能稳定的肾移植受者尿液中MCP-1的水平为(416±21)μg/L,对照组为(408±11)μg/L,两者无明显差 别。发生急性排斥反应的肾移植受者,尿液中MCP-1的水平为(1195±58)μg/L,明显高于对照组和肾功能稳定者(P<0.01)。经抗排斥治疗后,对治疗有反应的急性排斥反应受者,尿液中MCP-1的水平明显下降。结论:尿液中MCP-1的水平与肾移植急性排斥反应有密切关系。检测尿液中MCP-1的水平有助于急性排斥反应的诊断及预后判断。     

单个核细胞促血浆凝固活性监测急性排斥反应的实验研究

目的：探讨将外周血单个核细胞的促血浆凝固活性用于急性排斥反应监测的可能性。方法：将新西兰白兔随机分为３组：第１组做处体皮肤移植，第２，３组做异体皮肤移植，第３组于术前１ｄ至术后７ｄ皮下注射环孢霉素Ａ５ｍｇ（ｋｇ．ｄ）。术后每天观察衔以片的变化。     

猪人异种移植超急性排斥反应补体激活途径的探讨

异种移植最大障碍是由体液免疫造成的超急性排斥反应(Ｈｙｐｅｒａｃｕｔｅｒｅｊｅｃｔｉｏｎ,ＨＡＲ)。本实验用猪胸主动脉血管内皮细胞和人血清建立ＨＡＲ体外模型,ＬＤＨ释放法检测补体依赖的细胞毒作用,探讨ＨＡＲ中补体激活途径。1　材料与方法11　猪血管内皮细胞的...     

大鼠胰岛分离纯化及其小鼠肾被膜下移植排斥反应的研究

目的:探讨胰岛分离技术和异种胰岛移植排斥反应机制。方法:在控制不同条件的情况下,使用胰管内胶原酶灌注和不连续密度梯度纯化法获取大鼠胰岛,行小鼠肾被膜下移植,分别于术后当日和第3,5,7天取出移植物,分析排斥反应情况。结果:纯化后平均获取500+67个胰岛/胰腺,纯度约在70％～80％。统计学分析高糖和茶碱对胰岛刺激引起的胰岛索分泌,提示纯化后的胰岛功能良好。在未使用免疫抑制剂的情况下,植入的胰岛多在1周内完全排斥掉。术后5天,胰岛的形态已被破坏,淋巴细胞大量增多;术后第7天见不到完整的胰岛轮廓,仅见大量的淋巴细胞浸润。结论:严格控制各种实验条件,完善分离纯化技术,是优化胰岛获取的关键。大鼠胰岛小鼠肾被膜下移植中树突状巨噬细胞参与了排斥反应过程。     

肾移植患者尿液中供者细胞DNA变化在急性排斥反应诊断中的意义

目的：探讨肾移植受者尿液中供者细胞的出现与急性排斥反应的相关关系及其临床意义。方法：以供者为男性、受者为女性或HLA-DR抗原有错配的80例肾移植受者为研究对象，定期收集尿液标本，从中提取DNA，利用特异性引物聚合酶链反应分别检测Y染色体上特异的基因片段DYZ-1和HLA-DR抗原的基因DRB-1。结果：手术当天受者的尿液中即有供者细胞出现，随着时间的推移，尿液中供者细胞DNA的基因表达强度逐渐减弱，直至术后30天，仍有90．0％受者的尿液中有供者细胞DNA的基因表达，其中8例(29．6％)发生了急性排斥反应；出院后发生急性排斥反应者，90．0％的尿液标本中能检测到供者细胞DNA，抗排斥治疗结束后2周，83．3％转为阴性；肾功能良好的稳定期受者，仅6．7％的受者尿液中DYZ-1或HLA-DRB基因阳性。结论肾移植受者尿中供者细胞DNA的检测可以作为诊断急性排斥反应的一种方法，其基因表达强度变化为定量评价排斥反应提供了可能性。     

内皮微粒在肾移植急性排斥反应中的诊断作用

背景：血管内皮细胞的变化与移植排斥反应的关系极为密切,内皮微粒脱落于激活或凋亡的内皮细胞,能直接而特异地反映血管内皮细胞的变化,检测血浆中内皮微粒对肾移植排斥反应的诊断监测具有一定的理论和实际意义。 目的：探讨肾移植急性排斥反应时循环内皮微粒的数量和表型的变化及与急性排斥反应之间的关系。 方法：建立同基因和同种异基因大鼠腹腔原位肾移植模型;移植后5 d苏木精-伊红染色观察肾组织的病理学改变,并进行Banff评分;采用免疫组化法检测肾组织中细胞间黏附分子1表达;采用流式细胞术检测血浆中CD144⁺内皮微粒数量及细胞间黏附分子1⁺/CD144⁺内皮微粒的数量;分析内皮微粒的数量和表型与肾组织病理变化的关系。 结果与结论：与同基因移植组比较,异基因移植组Bnaff评分明显增加(P < 0.01),肾组织中细胞间黏附分子1表达明显增强(P < 0.01);与同基因移植组比较,异基因移植组CD144⁺内皮微粒的数量和携带细胞间黏附分子1的内皮微粒的水平明显增加(P < 0.01)。内皮微粒的数量与移植肾急性排斥反应的程度呈正相关(P < 0.01),携带细胞间黏附分子1内皮微粒的水平与移植肾脏中细胞间黏附分子1的表达呈正相关(P < 0.01)。提示肾移植后对内皮微粒数量和表型进行检测对诊断急性排斥反应的发生有一定意义。     

调节性T细胞在大鼠小肠移植急性排斥反应中的作用

目的 分析调节性T细胞在大鼠急性排斥反应中的作用。方法 采用免疫组化SABC染色法，测定BN→LEW大鼠小肠移植急性排斥反应时，外周血及移植肠浸润淋巴细胞中调节性T细胞：CD4^ ,CD8^ ,CD25^ T淋巴细胞及相关细胞因子IL－4和IFN－γ的表达，并与同基因大鼠间小肠移植（BN→BN）作比较。结果 外周血淋巴细胞分析显示，大鼠小肠移植急性排斥反应时，以CD4^ ,CD25^ ,T细胞为主，CD8^ 淋巴细胞只占少部分；分泌IL－4的细胞在术后4，7，14d分别只占14．3％，16．2％和16．9％。移植肠基底浸润的淋巴细胞以CD4^ ,CD25^＋和分泌IFN－γ的淋巴细胞为主。结论 在大鼠同种小肠移植中，急性排斥反应与CD25^ ,CD4^ T细胞及Th1相关细胞因子IFN－γ的表达增加相关。而CD8^＋T淋巴细胞和Th2相关细胞因子IL－4可能具有保护作用。     

Histopathologic changes in anti-angiotensin II type 1 receptor antibody-positive kidney transplant recipients with acute rejection and no donor specific HLA antibodies

Objective

To determine the association of antibodies against angiotensin II type 1 receptor (AT1R Ab) and histopathologic changes seen in patients with kidney allograft rejection and negative donor specific HLA antibodies (DSA).

Atypical cells in a voided urine cytology specimen in a renal transplant recipient

Voided urine is routinely collected from renal transplant patients to screen for polyomavirus. In rare cases, atypical lymphoid cells can be detected in voided urine and raise the suspicion of post‐transplant lymphoproliferative disorder (PTLD). However, further immunohistochemistry of the cell block and flow cytometry is frequently limited by the low cellularity and poor preservation of voided urine. Therefore, PTLD of the renal allograft is usually diagnosed from tissue biopsy or nephrectomy specimens. Herein, we report a rare case of atypical cells in a voided urine cytology specimen from a kidney transplant recipient. Needle core biopsy of the renal allograft showed monomorphic PTLD. Diagn. Cytopathol. 2017;45:69–72. © 2016 Wiley Periodicals, Inc.     

Short CommunicationThe MCP-1 Promoter -2518 Polymorphism in Behcet's Disease: Correlation Between Allele Types,MCP-1 Production and Clinical Symptoms among Korean Patients

Objective: To compare the G vs. A variation at the MCP-1 promoter -2518 position among normal Koreans and Behcet patients, and to investigate possible association of this polymorphism with disease pathogenesis.

Methods: The allele type of -2518 polymorphism was determined by restriction fragment length polymorphism of Pvu II. The level of MCP-1 in serum and culture supernatent was measured by sandwich ELISA.

Results: The average serum level of MCP-1 in Behcet patients was 2.37 times higher than in normal controls. The serum MCP-1 concentration was higher in G-allele carriers than in AA homozygotes, and symptoms accompanying graver cases of Behcet's disease, such as gastrointestinal inflammation and uveitis, were more frequent in patients with the G-allele. However, the frequency of G-allele in patient group was not higher than that in healthy Koreans, probably due to the dominance of G-allele in general Korean population. When stimulated in vitro with IL-1β and LPS, the mononuclear cells from patients carrying the G-allele showed a steeper increase in MCP-1 production than the boost observed in AA homozygotes.

Conclusion: Although the allele frequency of MCP-1 promoter -2518 polymorphism is not likely to be the reason for the elevated serum MCP-1 level in Korean patients with Behcet's disease, it is possible that proinflammatory factors induced in patients' serum cause stronger activation of MCP-1 expression from the G-type promoter, as well as increased incidence of uveitis and gastric ulcer, among carriers of the G-allele.     

Involvement of spinal monocyte chemoattractant protein-1 (MCP-1) in cancer-induced bone pain in rats

In this study, we examined the involvement of chemokine monocyte chemoattractant protein-1 (MCP-1) in the spinal cord of a rat model of cancer-induced bone pain (CIBP). In this model, CIBP was established by an intramedullary injection of Walker 256 cells into the tibia of rats. We observed a significant increase in expression levels of MCP-1 and its receptor CCR2 in the spinal cord of CIBP rats. Furthermore, the intrathecal administration of an anti-MCP-1 neutralizing antibody attenuated the mechanical allodynia established in CIBP rats. Likewise, an intrathecal injection of exogenous recombinant MCP-1 induced a striking mechanical allodynia in naïve rats. These results suggest that increases in spinal MCP-1 and CCR2 expression are involved in the development of mechanical allodynia associated with bone cancer rats.     

Differential diagnosis of acute allograft rejection and CMV-infection in renal transplantation by urinary cytology

Acute allograft rejection and CMV-infection are the most common complications after renal transplantation. Quick differential diagnosis between these two complications is still difficult but necessary, since both complications demand a different therapy. More than 2500 urinary samples from 33 transplanted patients were prospectively examined and part of them evaluated quantitatively. Urinary samples of patients with acute renal failure, long-term haemodialysis or immunosuppressive therapy served as controls. The following cytomorphological criteria proved to be useful: Tubular epithelial cells, casts, oxalate crystals (sand-glass shaped), dirty background, increasing erythrocyturia, mixed cell clusters, lymphocytes and mitoses. Rejection is going on when the number of renal tubular cells is increased and two or more further criteria are positive. 25 acute allograft rejections without acute renal failure were diagnosed clinically. All 25 rejections were also diagnosed by urinary cytology. Nevertheless, it is not possible to differentiate between acute allograft rejection and acute renal failure of other origin. CMV-infection was serologically detected in 7 patients. In 6 of them viral infected cells were found in the urine shortly after the onset of unspecific clinical symptoms. Besides the typical "owl-eye" cells milkglass nuclei, sometimes with eosinophilic condensation, were seen while criteria for transplant rejection were never observed at the same time. Cytologic examination of voided urine is a simple diagnostic help for the differentiation between allograft rejection and CMV-infection after renal transplantation.     

中国苏州地区汉族人群MCP-1基因多态性与急性胰腺炎相关性研究

目的探讨中国苏州地区汉族人群单核细胞趋化因子蛋白-1(monocyte chemoattractant protein-1,MCP-1)基因-2518A/G多态性与急性胰腺炎的相关性。方法采用聚合酶链反应-限制性片段长度多态性方法检测中国苏州汉族人群急性胰腺炎(acute pancreatitis,AP)患者101例[其中包括轻症急性胰腺炎(mildAP,MAP)78例,重症急性胰腺炎(servere AP,SAP)23例]和120名健康对照的MCP-1基因-2518位点基因型分布及基因频率,并评价基因多态性与AP易感性之间的相关性。结果对照组MCP-1-2518A/G位点的AA基因型频率较SAP组和MAP组高,差异有统计学意义(P<0.01),携AG及GG基因型者患MAP的风险度约是AA基因型的5.896倍(P<0.01,OR=5.896),患SAP的风险度约为7.011倍(P<0.05,OR=7.011)。而在MAP与SAP组间AA基因型频率差异无统计学意义(P=0.997)。对照组G等位基因频率明显低于MAP(P<0.01,OR=0.318)和SAP组(P<0.01,OR=0.309)。但G等位基因频率在MAP与SAP组间基因型分布差异无统计学意义(P=0.623,OR=1.211)。结论中国苏州地区汉族人群MCP-1-2518位点AA基因型可能有助于保护机体避免发生AP,G等位基因频率较高的人可能易患AP。但AA基因型和G等位基因频率不能预测SAP的发生。     

Diagnostic pitfalls in cytological diagnosis of subcutaneous fungal infection in renal transplant recipients

Renal transplant recipients (RTRs) are at increased risk of the development of a variety of skin infections that can result from graft-preserving immuno-suppressive therapy. In this study, we aimed to determine cytomorphological findings of fungal subcutaneous swelling in seven RTRs and to analyze diagnostic pitfalls in fungal cytology. A retrospective review of fine needle aspiration cytology (FNAC) smears of subcutaneous swelling with positive fungal elements in RTRs from 2008 to 2010 was performed. We had seven cases (all males; age range, 34-58 years, mean, 46.3 years). The time interval between the renal transplantation and appearance of swelling ranged from 8 to 19 months (mean, 13.4 months). The swelling was located on lower limb (six cases) and arm (one case). The lesion was solitary (six cases) and multiple (one case). The cytology of aspirated material showed branched septate fungal hyphae in six cases. These were well delineated on Periodic acid schiffs (PAS) and chromic silver methenamine (CSM) stains. One case showed presence of faint, thin walled, broad ribbon like hyphae. Culture of aspirated material was performed in four cases which grew phaeohyphomycosis in all. Histology of excised tissue showed numerous septate, branched, pigmented fungal elements suggestive of pheohyphomycosis in four cases and broad ribbon hyphae suggestive of zygomycosis in one case. All of our cases responded well with anti-fungal treatment. Fungal infection can manifest as subcutaneous swelling in RTRs. It is often severe, rapidly progressive and difficult to diagnose. FNAC is an important diagnostic tool which is simple, cost effective and rapid method.     

人静脉移植物中MCP-1的表达

目的：探讨人静脉移植物中单核细胞趋化蛋白-1（MCP-1）的表达及其与巨噬细胞浸润的关系。方法：应用免疫荧光组织化学技术对30个静脉移植物再塞标本中MCP-1、CD68（巨噬细胞的标记物）、CD3l（内皮细胞的标记物）的表达进行了检测，用激光共聚焦显微镜拍片，图片用Silicon Graphics Octane进行处理。结果：在正常静脉血管中，MCP-1表达很弱；外膜中有少量CD68免疫阳性细胞，中膜和内膜中少见；CD31免疫阳性细胞仅在血管腔内皮细胞和外膜小血管中可见。在病变静脉血管，MCP-1在内皮细胞、平滑肌细胞中的表达呈强阳性；CD68免疫阳性细胞在外膜、中膜和内膜均可见到；CD31免疫阳性细胞不仅出现在血管腔内皮细胞和外膜小血管内皮细胞，且在中膜小血管内皮细胞也大量出现。免疫双重染色显示内皮细胞和巨噬细胞均表达MCP-1。结论：人静脉移植物MCP-1的表达上调，且和巨噬细胞的浸润关系密切，提示MCP-l对静物移植物炎症细胞的浸润及新内膜的形成有非常重要的作用。