Kidney Transplantation in Patients with Antibodies against Donor HLA Class II

The immunologic risk associated with donor-specific antibodies (DSA) against Class II human leukocyte antigens (HLA) in kidney transplant (KTx) recipients is unclear. The aim of this study was to determine the outcome of KTx when DSA was detected only against HLA Class II. To isolate the impact of anti-Class II DSA, we retrospectively analyzed 12 KTx recipients who at baseline had a positive B-cell flow cytometric crossmatch (FXM) and a negative T-cell FXM. Using alloantibody specification analysis, 58.3% (7/12) had DSA against donor Class II and 41.7% had no demonstrable DSA. Biopsy-proven AMR occurred in 57% (4/7) in the Class II(+) group and 0% in the Class II(-) group (p > 0.05). Peritubular capillaries stained positive for C4d in 86% (6/7) of the Class II(+) patients and in 40% (2/5) of the Class II(-) patients (p > 0.05). One patient in the Class II(+) group lost their graft at 3 months to accelerated transplant glomerulopathy, while all other grafts were functioning 3-37 months posttransplant despite the persistence of anti-Class II DSA. We conclude that KTx recipients with clearly defined anti-Class II DSA are at risk for humoral rejection suggesting that desensitization and/or close posttransplant monitoring may be needed to prevent AMR.     

肝硬化大鼠肝部分切除术后肝再生的干预研究

目的　以肝硬化大鼠为动物模型 ,研究药物对肝硬化大鼠肝部分切除术后肝再生的影响。方法　取健康的Wistar雄性大鼠 6 4只 ,以 6 0 ?l4油溶液 0 .3ml/ 10 0 g皮下注射 ,同时饮用 5 %酒精溶液 ,4 5d后制成肝硬化动物模型。模型大鼠随机分为 4组 ,16只 /组。全麻下均行左、中叶肝切除术。术后各组按以下方案处理 :A组 (对照组 )注射生理盐水 1mg/ (kg·d) ,B组为泮托拉唑组 ,注射 0 .2mg/ (kg·d) ,C组为重组人生长激素组 ,注射 0 .5U/ (kg·d) ,D组为两药合用组 ,同时给予泮托拉唑注射 0 .2mg/ (kg·d) ,重组人生长激素注射 0 .5U/ (kg·d) ) ,连续给药 1周。抽取静脉血样 ,取肝脏组织 ,检测肝功能、有丝分裂指数 (MI)、增殖细胞核抗原 (PCNA)、细胞核DNA含量。结果　泮托拉唑组、重组人生长激素组、两药合用组MI、PCNA阳性染色细胞量、细胞核DNA含量均高于对照组 (P <0 .0 5 ) ,两药合用组MI、PCNA阳性染色细胞量、细胞核DNA含量均高于泮托拉唑组、重组人生长激素组 (P <0 .0 5 ) ,但各组间肝功能变化无明显差异。结论　泮托拉唑组及重组人生长激素均对肝硬化大鼠肝部分切除术后肝细胞再生有促进作用 ,两药联合应用肝细胞再生更明显 ,其详细机制须待进一步研究。     

Genetic immunization with GPI-anchored anthrax protective antigen raises combined CD1d- and MHC II-restricted antibody responses by natural killer T cell-mediated help

Studies have demonstrated that lipid rafts ultimately regulate the endocytosis of anthrax toxin via clathrin dependent pathway. Interestingly, GPI-anchored protein rich rafts have also been shown to be transported down to the endocytic pathway to reducing late endosomes. Taking advantage of this parallelism, we tried translating the anthrax toxin natural intoxication mechanism by administering a DNA chimera that encoded protective antigen attached to a mammalian GPI-anchor sequence at its C-terminus (pGPI-PA63). We also designed a chimera that had an additional N-terminal TPA leader sequence (pTPA.GPI-PA63) with an aim to target GPI-PA63 to ER where new CD1 molecules are synthesized. Analysis of antibody titers demonstrated successful priming and potential IgG titers after the first boost. In vitro cell proliferation studies in the presence of GPI-attached PA63 peptides revealed that there was a clonal expansion of CD4⁺ NK1.1⁺ helper T cell population which rapidly produced IL-4 in response to T cell receptor ligation. These cells provided direct B cell help that aided IgG formation. Effector responses generated by NKT cells were found to be MHC II-independent and CD1d-restricted. In addition, the group pTPA.GPI-PA63 also displayed low magnitude MHC-II restricted (CD1d-independent) NKT cell and CD4⁺ T cell helper responses in response to non-GPI attached PA63 peptides which overall resulted in the heightened responses seen for this group. Importantly, DNA vaccination mediated the generation of high avidity neutralizing antibodies that mediated protection against lethal toxin challenge.     

CD14+单核细胞人类白细胞抗原DR表达率与脓毒症关系的研究

目的了解烧伤延迟复苏时CDl4^＋单核细胞人类白细胞抗原DR（HLA—DR）表达率的变化，分析其与脓毒症的关系。方法选择烧伤面积大于30％TBSA的25例烧伤延迟复苏患者，于伤后1、3、7、14、28d取外周血，其中7例患者住院期间并发脓毒症，于脓毒症发生后连续2d亦取其外周血。另取20例健康体检者外周血作为对照。流式细胞仪检测CD14^＋单核细胞HLA．DR表达率，酶联免疫吸附测定法检测血浆中肿瘤坏死因子α（TNF—α）、白细胞介素10（IL-10）的浓度。结果非脓毒症患者伤后1、3、7、14、28dCD14^＋单核细胞HLA—DR表达率分别为（15±6）％、（74±5）％、（264±17）％、（284-16）％、（474-16）％，明显低于健康体检者[（924±10）％，P〈0．01]；脓毒症患者发生脓毒症后1、2d，该指标亦明显低于健康体检者及非脓毒症患者伤后1、7、14、28d（P〈0．01）。脓毒症患者TNF—d检出率及TNF—α、IL-10浓度，均高于非脓毒症患者和健康体检者（P〈0．05或P〈0．01）。伤后1、7、28d，外周血CD14^＋单核细胞HLA—DR表达率与IL—10浓度呈显著负相关（r分别为-0．9963、-0．7459、-0．8474，P〈0．01）。结论烧伤延迟复苏患者免疫功能低下，促炎性介质释放量增加，并发脓毒症时则更为严重。外周血CD14^＋单核细胞HLA—DR表达率可作为动态检测患者免疫功能状态的有效指标。     

人胎冠状动脉原位杂交c-myc和jun原癌基因表达

目的研究原癌基因c-myc和jun在人胎冠状动脉发育过程中的表达与平滑肌细胞增殖的关系.方法用原位杂交方法检测,胎龄分别为16周、22周(因治疗需要引产)的胎儿和意外死亡的足月胎儿冠状动脉前降支c-myc mRNA和jun mRNA的表达水平.杂交反应产物用图像分析仪(MIAS300)作定量分析.结果C-myc mRNA原位杂交反应产物与被测血管区域面积的百分比在16周、22周和足月胎儿分别是70、56和10;Jun mRNA的杂交信反应产物与被测血管区域面积的百分比在这三个时期分别是68、53和8.两个原癌基因在不同阶段的表达均具有显著性差异.结论本实验首次报道c-myc和jun在人胎冠状动脉发育过程中平滑肌的表达图型,c-myc和jun在胎儿冠状动脉平滑肌细胞增殖和内膜的形成过程中可能具有重要的调控作用.     

外周血人类斯钙素1基因表达与消化道恶性肿瘤微转移关系的研究

目的：探讨靶基因人类斯钙素1(hSTC—1)在消化道恶性肿瘤患者外周血中的表达以及与肿瘤微转移的关系。方法：采用RT—PCR方法检测69例消化道恶性肿瘤患者外周血中的人类斯钙素1(hSTC—1)mRNA，12例健康成人、4例妊娠期妇女、14例消化道炎症疾病患者，并检测12例消化道恶性肿瘤患者手术中新鲜组织标本中的hSTC—1 mRNA。结果：69例消化道恶性肿瘤患者外周血中，食管癌组hSTC—lmRNA的阳性率为52．9％(9／17例)，胃癌组的阳性率为57．7％(15／26例)，大肠癌组的阳性率为53．8％(14／26例)，消化道恶性肿瘤患者手术中新鲜组织标本的阳性率为100％(12／12例)；而健康成人、妊娠期妇女、消化道炎症疾病患者外周血中无1例出现阳性。将肿瘤患者hSTC-1 mRNA检测结果与临床病理结合分析发现：外周血中的hSTC-1m RNA的表达与肿瘤的浸润深度、临床病理分期、淋巴结转移有显著相关性。结论：应用RT—PCR方法检测消化道恶性肿瘤患者外周血中的hSTC—1m RNA具有较高的敏感性和特异性，可成为检测肿瘤外周血微转移的一个新指标。     

人血清载脂蛋白A-I的分离、纯化及其测定

用超离心法制备人血清高密度脂蛋白(HDL);甲醇/氯仿脱脂;经Sephadex G-150层析及DEAE-纤维素(DE-52)离子交换得纯化载脂蛋白A-I(Apo A-I),免疫新西兰兔,获得效价为1:32的特异性抗人Apo A-I抗血清。用本室建立的单向火箭免疫电泳法测定了158名健康中年人与9名老人的血清Apo A—I含量,结果发现:β-脂蛋白<5g/L,总胆固醇<2mmol/L的人血清Apo A-I含量男女分别为2.06±0.16g/L和2.07±0.15g/L;而β脂蛋白≥5g/L,总胆固醇≥2mmol/L的人血清Apo A-I含量男女分别为1.75±0.15g/L和1.82±0.1g/L,由此可见,血脂正常的男性和女性的Apo A—I含量均显著高于血脂高的男性和女性。80岁以上老年人的血清Apo A-I含量为2.1±0.1g/L,也在正常范围。与文献报道比较,我国人的血清Apo A-I含量高于外国人,这与我国人HDL水平较外国人高一致。     

沙培林增强人腹腔巨噬细胞抗癌免疫功能的研究

目的:探讨腹腔内注射沙培林增强人腹腔抗癌免疫功能的机制。方法:72例早中期胃肠道肿瘤患者术前48h和24h腹腔内分别注射生理盐水和5KE的沙培林,术中采集腹腔内巨噬细胞,计数并测定乳酸脱氢酶(LDH)和酸性磷酸酶(ACP)的活性,巨噬细胞吞噬活力,一氧化氮(NO)的分泌以及对人胃癌MKN1细胞的细胞毒性进行分析。同时采集大网膜,对大网膜乳斑的数量和面积进行观察。结果:沙培林显著增加腹腔巨噬细胞(PMΦ)的数量和NO的分泌,增强LDH和ACP的活性,吞噬活力,以及抗癌细胞毒性,也显著增加了大网膜乳斑的数量和面积。结论:腹腔内注射沙培林可显著增加人大网膜乳斑的数量和面积,并因此增加PMΦ的数量,增强PMΦ的活性。因而增强了腹腔巨噬细胞的免疫功能。     

The effect of sucralfate,an agent for gastroprotection on the healing of split thickness skin graft donor sites

Skin is the most commonly used tissue for the transplantation. A meticulous care of the donor site is needed to prevent scarring, delayed healing, and pain. Various agents and dressing materials have been reported to help healing of skin graft donor sites. Sucralfate is an extensively used agent, which provides acute gastroprotection and acceleration of chronic ulcer healing. In this study, we assessed the effects of topical sucralfate on the healing of the split thickness skin graft donor sites in a prospective comparative way. The study was carried out on 32 randomly chosen patients undergoing surgery for various causes and requiring split thickness skin graft resurfacing. The upper halves of the skin graft donor sites on the thighs were simply covered with paraffin gauze and the lower half was covered with sucralfate-soaked paraffin gauzes. The day of full epithelization varied from 6 to 9 days and 8–12 days on the sucralfate-applied areas and on the control sites, respectively. The mean value of the healing was 7.01 days in the studied lower halves and 10.8 days in the upper halves. The healing rate was strikingly faster and less painful on the sucralfate-applied areas. We were able to discharge patients earlier than usual, and patients’ comfort increased. It seems that sucralfate is a promising topical agent to increase the healing rate and decrease the incidence of associated problems such as pain and hypertrophic scar.     

Permanent prostate brachytherapy for Japanese men: Results from initial 100 patients with prostate cancer

OBJECTIVE: To evaluate the initial results of brachytherapy for prostate cancer with permanent iodine-125 implant in Japan. METHODS: The results obtained with brachytherapy in the initial 100 Japanese patients treated at Nagano Municipal Hospital were reviewed. Patients with a prostate-specific antigen (PSA) level of less than 10 ng/mL and a Gleason's scores of 5, 6, 3 + 4 were classified as having a low risk of recurrence. Patients with a PSA level of 10-20 ng/mL and/or a Gleason's score of 4 + 3 were classified as having an intermediate risk for recurrence. Seventy-eight of the low-risk patients and 19 of the intermediate-risk patients were treated by seed implants alone, or seed implants combined with preceding external radiation, respectively. A total of 53 patients received neoadjuvant hormone therapy. The efficacy and morbidity of brachytherapy were investigated using the serum PSA, International Prostate Symptom Score, quality of life score and uroflowmetry data. RESULTS: The average V100 and D90 obtained by post-implant dosimetry was 94.3 and 113.7%, respectively. Serum PSA decreased gradually after treatment, although it had still not reached a nadir after 1 year. There was little difference of the PSA level between the patients with and without neoadjuvant hormone therapy even at 1 year after seed implantation. There were no PSA biochemical failure or clinical recurrence during the follow-up period. Voiding symptoms worsened until 3 months after treatment, and then gradually improved. Acute urinary retention occurred transiently in one patient (1%). Rectal bleeding and severe diarrhea did not occur. CONCLUSION: Brachytherapy is a feasible and effective option for the treatment of prostate cancer in Japanese men. Brachytherapy may have a different effect in Japanese patients with respect to voiding symptoms. Urinary retention was rare, but voiding symptoms were persistent in Japanese patients. Neoadjuvant hormone therapy deserves investigation to determine whether it can achieve better results, especially in patients with an intermediate risk.