嵌合体的定量检测在异基因造血干细胞移植术后过继免疫治疗中的应用

目的探究供体细胞嵌合率（DC）的动态定量检测在预测异基因造血干细胞移植（allo-HSCT）后的转归,确定过继免疫治疗（AIT）时机和评价AIT疗效中的作用。方法采用复合扩增荧光标记短串联重复片段（STR）一PCR结合毛细管电泳方法,对84例allo—HSCT患者骨髓或外周血中DC进行定量检测,并对其中接受AIT的16例患者治疗前后的DC进行连续定量检测。结果移植后84例患者中有22例出现受体细胞比例增高的混合嵌合（MC）状态,DC均低于90％,6例MC患者未接受供体淋巴细胞输注（DLI）治疗,结果均死于血液学复发,另外16例患者接受AIT治疗,其中复发和植入失败的12例患者在出现临床症状之前DC均出现大幅度下降（24．8％-86．2％）。16例中11例出现治疗反应（有效率68．8％）。治疗有效患者在AIT后短期内出现DC回升（DC≥90％）,供体细胞占优势,并转化为完全供体细胞嵌合状态（6／11）或稳定的MC状态（5／11）,临床上出现移植物抗宿主病表现。无效患者AIT后DC均未稳定回升,3例首次DLI无效的患者接受了第二次DLI治疗。结论DC的动态定量检测可早期发现复发或植入失败的高危患者,尽早实施AIT,可提高发生细胞遗传学或分子生物学复发患者的疗效。此外DC的定量检测还可预测DLI的临床疗效,对首次DLI治疗无效的患者后继的AIT提供重要的参考依据。     

甲磺酸伊马替尼治疗异基因造血干细胞移植后复发的慢性粒细胞白血病五例临床观察

异基因造血干细胞移植（allo-HSCT）是目前惟一能治愈慢性粒细胞白血病（CML）的方法。白血病复发仍是导致移植失败的主要原因之一。在慢性期接受allo-HSCT患者的复发率较低（5％～20％），但在疾病加速期或急变期进行移植，复发率则明显升高（30％％～60％）。移植后复发的治疗主要包括供者淋巴细胞输注（DLI），IFNα和第二次移植。虽然DLI可诱导长期的分子学完全缓解（CR），已成为治疗allo-HSCT后复发CML患者的首选方法，但同时可引起移植物抗宿主病（GVHD），骨髓抑制，甚至死亡。     

HBsAg阳性患者异基因造血干细胞移植后的临床研究

目的 探讨HBsAg阳性患者接受异基因造血干细胞移植(allo-HSCT)后的肝脏不良事件的发生及影响该类患者长期存在的危险因素.方法 回顾性分析本所2001年3月至2006年11月接受allo-HSCT治疗的26例HBsAg阳性患者的临床资料.造血干细胞来源:18例为HLA配型相合同胞,7例为HLA配型不合亲缘供者,1例为无关供者.所有患者移植前后均排除丙型肝炎病毒感染.2例移植前HBV DNA阳性,其余均表达阴性.结果 26例患者急性移植物抗宿主病(Agvhd)累积发生率为50.0%.在可评估的20例中,5例发生了慢性GVHD,累积发生率25.0%.移植后出现肝功能异常共有15例(57.7%).出现肝功能受损的病因主要为:乙型肝炎及肝脏GVHD.移植后乙型肝炎再燃5年累积发生率33.4%,发生中位时间为82(65～159)d.拉米夫定预防性应用13例,1例发生乙型肝炎事件;未给药组为11例,7例发生乙型肝炎事件,两者累积发生率差异有统计学意义(P=0.006);预防用药组未发生一例肝衰竭,未预防用药组发生4例肝衰竭.死亡原因:5例肝衰竭,3例肺部感染,2例移植后疾病复发.26例患者中共有10例死亡,5年生存率(OS)为59.0%.多因素分析最终确定与OS发生相关的危险因素为肝衰竭(P=0.000).结论 HBsAg阳性患者接受allo-HSCT治疗后常出现肝功能异常表现,其首要病因为乙型肝炎,所导致的肝衰竭严重影响患者预后.而对HBsAg阳性患者预防性应用拉米夫定能够有效预防移植后乙型肝炎的复燃.     

供者淋巴细胞输注对恶性血液病异基因造血干细胞移植后复发的治疗作用

异基因造血干细胞移植(Allo HSCT)已成为治疗恶性血液病的主要方法之一,但移植后恶性血液病的复发降低了患者的长期生存率。目前Allo HSCT后复发的治疗包括停用免疫抑制剂、供者淋巴细胞输注(DLI)、二次移植、化疗、细胞因子治疗等方法,而DLI是其中疗效较为显著,毒性相对较小,作用比较肯定的方法,并且已作为非清髓造血干细胞移植(NST)后续治疗的组成部分[1],甚至在自体移植中也使用DLI[2]。本文就DLI对Allo HSCT后恶性血液病复发治疗的相关问题作一综述。DLI诱发移植物抗白血病(GVL)效应的机制在HSCT后发生移植物抗宿主宿(…     

非清髓性造血干细胞移植加供者淋巴细胞输注治疗多发性骨髓瘤1例报告并文献复习

目的研究非清髓性造血干细胞移植(NST)治疗多发性骨髓瘤(MM)的疗效,观察移植相关并发症的发生.方法1例42岁MM患者,供者为其胞姐,HLA配型完全相合.动员方案粒细胞集落刺激因子(G-CSF)10 μg·kg-1·d-1×5 d.预处理方案抗胸腺细胞球蛋白(ATG)8 mg·kg-1·d-1×3 d,马法兰(MEL)120 mg/m2×1 d.移植单个核细胞数(MNC) 6.5×108/kg;CD34+细胞 4.4×106/kg.环胞菌素A(CsA)和短程甲氨蝶呤(MTX)预防移植物抗宿主病(GVHD).移植后分别于+41 d、+76 d和+112 d进行3次供者淋巴细胞输注(DLI).结果移植后15 d中性粒细胞计数> 0.5×109/L,21 d血小板计数>50×109/L,24 d性染色体和微卫星法DNA指纹图监测显示为混合嵌合体,随着DLI的进行,逐渐转为供者型完全嵌合体,骨髓瘤细胞和血清M蛋白均逐渐消失,移植后8个月达完全缓解.在+180 d(第三次DLI后68 d)发生II度急性GVHD,经甲泼尼龙和CsA治疗得以控制.现随访36个月,患者情况良好,仍处于完全缓解状态.结论非清髓性造血干细胞移植加供者淋巴细胞输注治疗MM是可行和有效的.     

供者淋巴细胞输注治疗异基因造血干细胞移植后EB病毒相关的淋巴细胞增殖性疾病的疗效和安全性

目的 采用控制数量的供者淋巴细胞输注(DLI)治疗EB病毒(EBV)相关的淋巴细胞增殖性疾病(PTLD),观察其有效性和安全性.方法 2006年11月到2009年11月移植的患者,临床或病理诊断EBV相关的PTLD采用DLI治疗者纳入研究.采用COBE血球分离机应用淋巴细胞程序采集白细胞,首选原供者,次选直系亲缘供者,冻存外周血采集物(G-PB)与移植时输入的外周血干细胞(PBSC)组分相同.控制输入的单个核细胞(PBMC)在(0.5～1.0)×108/kg.原基础免疫抑制剂预防移植物抗宿主病(GVHD),观察疗效及其副作用.结果 PTLD患者9例,输注13例次,输入外周血PBMC 0.8(0.16～1.03)× 108/kg,CD3+T淋巴细胞4.2(1.6～5.7)×107/kg.7例半相合移植患者采用了原供者,有效率为7/7,完全缓解率为6/7,退热中位时间2(1～5)d,淋巴结缩小中位时间6(1～14)d;7例中6例发生GVHD,均为轻中度,得到控制.至今无病存活3例.2例非亲缘HSCT采用了半相合供者的外周血白细胞,仅获得短暂部分疗效.结论 在配型不合/半相合移植中,采用原供者控制细胞数量的DLI并免疫抑制剂预防GVHD治疗EBV相关的PTLD安全有效.最佳方案尚需进一步研究.     

异基因造血干细胞移植术后复发防治策略

移植后复发是导致异基因造血干细胞移植(allo-HSCT)病患死亡的首要原因。在预处理方案中应用氟达拉滨、氯法拉滨等新型化疗药物和分子靶向药物、移植后利用高度敏感的分子检测技术有效监测微小残留病(MRD)、基于MRD监测的预防性与治疗性供者淋巴细胞输注(DLI)及DLI结合新型化疗药物或靶向药物等新的预防与治疗策略有望进一步减少移植后复发和提高复发患者存活率。     

难治性白血病异基因骨髓移植后髓外复发的危险因素和治疗

目的：探讨难治性白血病移植后髓外复发（软组织）的危险因素、治疗方法和疗效。方法：2001～2003年间15例急性白血病患者进行了异基因造血干细胞移植（allo-HSCT），其中6例为难治性或晚期白血病。6例晚期患者中4例移植后出现髓外复发（软组织肉瘤，EMD），治疗全部采用联合化疗加放疗、供者淋巴细胞输注（DLI），2例加用免疫细胞因子，1例进行了2次移植。结果：4例患者移植到发生EMD的间隔时间分别为10、5、8、8个月，均无骨髓复发的证据。经治疗2例无病生存，1例带瘤生存，1例死亡。allo-HSCT后生存期分别为15、27＋、24＋、19＋个月。结论：①移植后出现髓外复发患者的预后较差；②EMD的发生与移植前疾病状态、有无移植物抗宿主病（GVHD）、移植后免疫抑制剂的停用时间有关。晚期患者移植后，免疫抑制剂在可能的情况下应尽早减量、停药；③髓外复发早期，化疗和DLI在部分病例中有效；而当肿瘤负荷较大时，DLI的疗效欠佳，此时降低预处理剂量的二次移植不失为一种有效的治疗选择。     

HLA配型相合的造血干细胞移植治疗重型再生障碍性贫血的临床研究

目的 评价HLA配型相合的异基因造血干细胞移植(allo-HSCT)治疗重型再生障碍性贫血(SAA)的疗效.方法 2000年1月至2008年11月采用allo-HSCT治疗SAA患者20例,其中同胞相合移植17例,非血缘关系移植3例.预处理采用环磷酰胺(Cy)50 mg·kg~(-1)·d~(-1)4 d加抗淋巴细胞免疫球蛋白(ATG)2.5 mg·kg~(-1)·d~(-1)或20 mg·kg~(-1)·d~(-1) d.移植物抗宿主病(GVHD)的预防方案为经典的环孢素A(CsA)联合短程甲氨蝶呤(MTX)及霉酚酸酯(MMF).同胞供者采集经重组人粒细胞集落刺激因子(G-CSF)动员的骨髓及外周血干细胞,非血缘供者单纯采集外周血干细胞.结果 回输单个核细胞中位数为7.89(4.00-14.21)×10~(8)/kg,所有患者均获供者造血重建,粒细胞植活中位时间14(11～20)d;血小板植活中位时间12(8～108)d.但1例患者发生晚期排斥,行另一供者二次移植后植活.21例次移植后共发生6例次急性GVHD(I度3例,Ⅱ度皮肤3例),发生率16%.19例生存期>100 d的患者中有7例发生慢性GVHD,其中4例为局限型,3例为广泛型.截至2009年2月28 日,经过中位18(2.0～106.8)个月的随访,共有17例患者无病生存,总生存率为82.5%.结论 采用Cy+ATG的预处理方案对SAA患者进行HLA配型相合HSCT,植活率高,可以获得良好的疗效.     

异基因造血干细胞移植后单独胃肠道受累急性移植物抗宿主病的临床特征

目的 分析异基因造血干细胞移植(allo-HSCT)后患者发生单独累及胃肠道的急性移植物抗宿主病(aGVHD)的临床特征.方法 回顾性总结2007年11月至2008年12月在我院行allo-HSCT治疗的307例血液病患者的临床资料,分析单独胃肠道受累的aGVHD的临床特征、危险因素、治疗效果及预后.结果 307例患者中174例(56.7%)发生aGVHD,其中单独累及胃肠道的38例,占21.8%.单独累及胃肠道的aGVHD的发生与受者性别、年龄、供受者性别关系、供受者血型关系、基础疾病诊断及回输的单个核细胞均无关;但与移植类型相关,亲缘全相合组发生率高于亲缘不合组及非血缘组.单独累及胃肠道的aGVHD患者经糖皮质激素或抗CD25单克隆抗体治疗,仅有2例死于aGVHD未控制合并感染,总有效率达94.7%;发生Ⅱ～Ⅳ度aGVHD但非单独累及胃肠道的患者抗aGVHD治疗总有效率为85.2%;组间比较差异有统计学意义(P=0.015).单独累及胃肠道的aGVHD中有12例应用抗CD25单克隆抗体治疗,治疗均有效.结论 HSCT后单纯累及胃肠道的aGVHD并不少见,其发生率在亲缘全相合组高于亲缘不全相合组及非血缘组,单独累及胃肠道的aGVHD对抗aGVHD治疗效果好,可能与应用抗CD25单克隆抗体疗效较好有关.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.