骨髓增生异常综合征骨髓切片中造血细胞凋亡的初步研究

目的 :观察骨髓增生异常综合征 (MDS)骨髓塑料包埋切片中原位细胞凋亡状况。方法 :用 DNA末端原标记 (ISEL)方法检测 33例 MDS患者骨髓塑料冷包埋切片中凋亡细胞 ,缺铁性贫血 15例作对照 ,并对 MDS之 ISEL 阳性细胞经常规染色光学显像镜分析细胞来源。结果 :33例 MDS的凋亡细胞平均为 (38.0 6± 2 4.6 1)个 /m m2 ,对照组为 (19.2± 7.98)个 /m m2 ,P <0 .0 1,差异有极显著性意义。 MDS中低危组 (RA+RAS)凋亡率(43.5 8± 39.40 )个 /mm2 较高危组 (RAEB+RAEB- T) (2 6 .0 4± 2 1.0 2 )个 /mm2 高 (P <0 .0 1) ,差异有极显著性意义。ISEL 阳性细胞主要来源于粒、红、巨三系造血细胞 ;分别占 38.45 % ,2 7.33% ,7.87%。结论 :MDS存在过度凋亡 ;MDS之 RA+RAS组凋亡高于 RAEB+RAEB- T组 ;凋亡细胞主要来源于三系造血细胞。     

骨髓增生异常综合征T细胞早期激活及可溶性肿瘤坏死因子受体的研究

目的　研究骨髓增生异常综合征 (MDS)外周血CD+ 4 、CD+ 8T细胞早期激活标志CD69的表达及血清、骨髓可溶性肿瘤坏死因子受体 1、2 (sTNF R1、2 )的水平及其意义。方法　在植物血凝素 (PHA) 2 0mg/L条件下进行全血细胞培养 ,于 0h和 4h分别用流式细胞仪对CD+ 4 、CD+ 8T细胞CD69的表达进行分析。用ELISA法检测血清和骨髓sTNF R1、2的水平。结果　PHA刺激前难治性贫血 (RA)与难治性贫血伴环形铁粒幼细胞增多 (RAS)CD+ 4 、CD+ 8细胞CD69的表达率分别为 8 32 %、9 88% ,难治性贫血伴原始细胞增多 (RAEB)与转变中的RAEB(RAEB T)CD+ 8细胞CD69的表达率为7 92 %。PHA刺激后MDS患者CD+ 4 、CD+ 8细胞表达CD69明显增强 ,RA +RAS为 5 3 4 6 %、5 1 6 3% ;RAEB +RAEB T为 4 2 93%、4 1 96 % ,CD+ 4 与CD+ 8细胞CD69的表达率相似。MDS两种sTNF R1水平均明显升高 ,RA +RAS组sTNF R1血清为 (1 5 8± 0 6 8) μg/L ,骨髓为 (2 10± 0 2 6 ) μg/L ;sTNF R2血清为 (1 4 1± 0 5 0 ) μg/L ,骨髓为 (1 95± 0 6 4 ) μg/L ;RAEB +RAEB T组sTNF R1血清为 (2 6 2± 2 5 5 ) μg/L ,骨髓为 (3 12± 0 6 7) μg/L ;sTNF R2血清为 (1 96± 0 5 6 ) μg/L ,骨髓为(3 0 9± 0 6 2 ) μg/L。血清sTNF R2水平与PHA刺激     

Fas-Fasl系统及凋亡与再生障碍性贫血关系的研究

目的 :研究Fas Fasl系统及凋亡与再生障碍性贫血 (Aplasticanemia ,AA)发生、发展的关系。方法 :分别用TUNEL方法和免疫组织化学标记方法检测凋亡细胞和Fas、Fasl表达细胞 ,并计算其百分率 ;用ELISA方法检测血清中的sFas含量。结果 :AA患者骨髓单核细胞 (BMMNC)中的凋亡细胞百分率为 2 9.6 3%± 17.71% ,Fas表达细胞百分率为 2 4 .81%± 9.4 5 % ,Fasl表达细胞百分率为 2 1.83%± 7.91% ,血清sFas含量为 5 .72± 0 .34μg/L ,与正常对照组 (分别为 2 .2 7%± 0 .74 %、1.93%± 0 .32 %、4 .18%± 1.0 2 %及 7.5 5± 0 .79μg/L)比较差异均有显著性意义 (P <0 .0 5 ) ;AA患者凋亡细胞百分率与Fas+ 细胞或Fasl+ 细胞百分率之间无直线相关关系 (P >0 .0 5 )。结论 :Fas Fasl系统介导的凋亡以及机体对其调控的失常可能参与了AA患者骨髓造血衰竭的病理生理过程 ,但体内可能还存在另外的机制 ,可造成AA患者BMMNC凋亡增加     

老年非小细胞肺癌患者外周血T淋巴细胞活化抗原表达的临床研究

目的　观察老年非小细胞肺癌患者外周血T淋巴细胞活化抗原CD3+ /CD2 5 + 、CD3+ /HLA DR+ 和CD3+ /CD6 9+ 的表达。　方法　应用流式细胞双色免疫荧光标记术对 4 5例老年肺癌患者上述 3种外周血T淋巴细胞活化抗原表达进行检测 ,并与非老年肺癌组、健康老年对照组和老年良性病变组对比。　结果　 4 5例老年肺癌患者外周血T淋巴细胞中CD3+ /CD2 5 + 、CD3+ /HLA DR+ 和CD3+ /CD6 9+ 表达 (分别为 7 2 4± 1 85、2 8 4 6± 5 39和 7 78± 2 6 3)低于健康老年对照组(分别为 10 35± 2 5 4、37 16± 5 5 1、11 0 2± 2 18)和良性病变组 (分别为 9 5 3± 3 0 2、35 33± 5 2 3、10 6 7± 2 4 5 ) ,P <0 0 1。老年组与非老年组表达水平比较差异有显著性 (P <0 0 5或P <0 0 1)。对照组和老年良性病变组之间差异无显著性 (P >0 0 5 )。Ⅲ、Ⅳ期 (分别为 7 15± 1 13、2 5 32±5 2 3、7 14± 2 81)和Ⅰ、Ⅱ期 (分别为 8 0 6± 1 2 1、30 2 7± 6 0 5、8 4 3± 2 6 7)表达比较 ,差异有显著性 (P <0 0 1)。中、高分化程度的肺癌患者其表达 (分别为 8 2 3± 1 2 3、34 0 5± 6 6 1、11 2 5± 3 2 2 )明显高于低分化程度的肺癌 (分别为 7 0 2± 1 35、2 5 31± 5 87、5 31± 3 5 7) ,P <0 0     

二硝基氟苯实验性结肠炎模型的建立及T抑制细胞亚群的研究

目的　探讨CD8+ T细胞亚群尤其是CD8+ CD2 8-T抑制细胞在 2 ,4 二硝基氟苯 (DNFB)大鼠实验性结肠炎模型中的变化及其可能作用。方法　建立DNFB实验性结肠炎大鼠模型 ,流式细胞仪计数检测脾脏和结肠黏膜上皮细胞内CD8+ T细胞、CD8+ CD2 8+ T细胞和CD8+ CD2 8-T抑制细胞的变化。结果　成功地建立了DNFB大鼠实验性结肠炎模型。模型 (n =12 )中脾脏和结肠黏膜上皮细胞内CD8+ T细胞含量与对照组 (n =8)差异无显著性 [脾脏 :(34.6± 7.2 ) %比 (33.5± 9.4 ) % ,结肠 :(14 .0± 8.9) %比 (18.0± 4 .1) % ,P >0 .0 5 ],CD8+ CD2 8-T抑制细胞含量均显著高于对照组 [脾脏 :(11.3± 2 .3) %比 (5 .6± 1.0 ) % ,结肠 :(6 .5± 5 .4 ) %比 (1.1± 0 .6 ) % ,P <0 .0 5 ],而结肠黏膜上皮细胞内CD8+ CD2 8+ T细胞含量显著低于对照组 [(7.5± 4 .2 ) %比 (16 .9± 4 .1) % ,P <0 .0 5 ],脾脏CD8+CD2 8+ T细胞含量亦低于对照组 [(2 3.3± 6 .1) %比 (2 7.8± 9.7) % ,P >0 .0 5 ]。实验性结肠炎模型中脾脏和结肠黏膜上皮细胞内CD8+ CD2 8-T抑制细胞占CD8+ T细胞的比例均显著高于对照组 [脾脏 :(33.3± 5 .5 ) %比 (18.4± 7.3) % ,结肠 :(46 .0± 14 .3) %比 (6 .1± 3.7) % ,P <0 .0 5 ]。结论　DNFB可诱导大鼠实验     

人骨髓基质干细胞的分子表型特征及微环境依赖向内皮分化的能力

目的 :探讨人骨髓基质干细胞的分子表型特征及与成熟内皮共培养时向内皮分化的能力。方法 :采用密度梯度离心法分离培养人骨髓基质干细胞 ,用荧光激活细胞分选法分析其CD34、CD10 5和CD16 6表达率 ,用免疫荧光细胞化学法观察与成熟兔主动脉内皮共培养的人骨髓基质干细胞Flk -1和vWF蛋白表达 ,并分析anti VEGF抗体对骨髓基质干细胞Flk -1表达的影响。结果 :分离培养的骨髓基质干细胞CD34表达率为 (4 .16± 0 . 16 ) % ,与阴性对照 (4. 0 6± 0 . 2 3) %相比无统计学差异 ,CD10 5表达率为(90 .2 0± 2 . 35 ) % ,CD16 6表达率为 (82. 30± 3. 2 2 ) % ,均明显高于阴性对照 (n =6 ,P <0 .0 5 ) ;与成熟内皮细胞共培养 5d时 ,vWF染色仍为阴性 ,但部分骨髓基质干细胞开始表达Flk -1;anti VEGF抗体呈浓度依赖地抑制Flk- 1阳性骨髓基质干细胞计数 (n =6 ,P <0 .0 5 )。结论 :与成熟内皮共培养的骨髓基质细胞具有微环境依赖向内皮细胞分化的能力。     

骨髓增生异常综合征患者骨髓凋亡细胞类型及TRAIL表达水平研究

目的探讨骨髓增生异常综合征(MDS)患者骨髓细胞凋亡累及的细胞类型,以及其与肿瘤坏死因子相关凋亡诱导配体(TRAIL)表达水平的关系。方法用流式细胞术检测骨髓有核细胞总凋亡率、CD3+、CD3+4细胞凋亡率和TRAIL表达水平,运用CellQuest及ModFit软件对所测得结果进行数据分析。结果MDS患者骨髓有核细胞总凋亡率为(17.39±10.17)%,CD+3、CD3+4细胞凋亡率分别为(20.41±11.61)%、(21.32±12.17)%,TRAIL表达水平为(7.55±3.10)%,均显著高于正常对照组[(5.88±1.38)%、(5.47±1.65)%、(4.92±2.11)%、(1.26±0.46%)],P<0.05。早期MDS有核细胞总凋亡率为(21.95±9.44)%、TRAIL表达水平为(9.00±2.47)%,均高于进展期[(8.26±2.27)%、(4.65±2.04)%],P<0.05。骨髓有核细胞总凋亡率与骨髓原始细胞数呈负相关(r=-0.65,P<0.05),与TRAIL表达水平呈正相关(r=0.85,P<0.05)。结论MDS存在过度凋亡,凋亡累及早期造血细胞和淋巴细胞;TRAIL可能是引起凋亡增加的原因之一。     

支气管肺泡灌洗液T淋巴细胞亚群和细胞DNA含量分析对肺癌的诊断价值

目的　探讨支气管肺泡灌洗液T淋巴细胞亚群和细胞DNA含量分析对肺癌的诊断价值。方法　用流式细胞仪对 3 0例肺癌和 12例肺良性病变患者的支气管肺泡灌洗液 (BALF)作T淋巴细胞亚群和细胞DNA含量分析 ,并与纤维支气管镜活检和刷检比较。结果　中央型肺癌组BALF中CD3 + (11.473± 3 .677) %、CD4+ (3 .660± 1.60 5 ) %、CD8+ (2 .3 49± 0 .880 ) %和CD4+ /CD8+ (1.411± 0 .3 0 9) ,周围型肺癌组BALF中CD3 + (12 .2 73± 4.42 5 ) %、CD4+ (2 .897±0 .695 ) %、CD8+ (2 .2 5 9± 0 .5 91) %和CD4+ /CD8+ (1.3 0 7± 0 .2 45 ) ;肺良性病变组BALFCD3 +(2 5 .0 78± 7.13 8) %、CD4+ (13 .2 44± 4.15 9) %、CD8+ (7.63 1± 3 .713 ) %和CD4+ /CD8+ (2 .2 78±0 .619) ,前两组显著低于后组 (肺良性病变组 ) (P <0 .0 0 1)。以异常二倍体为阳性标准 ,诊断肺癌的敏感性为 83 % ,特异性为 92 % ;中央型肺癌异常二倍体阳性率为 89% ,与活检 (95 % )和刷检(68% )比较 ,差异无显著性 (P >0 .5和P >0 .1) ;周围型肺癌异常二倍体阳性率为 82 % ,显著高于活检 (2 7% )和刷检 (2 7% )阳性率 (P均 <0 .0 5 )。结论　肺癌患者免疫功能低下 ,是肿瘤发生发展的原因之一 ,支气管肺泡灌洗液细胞DNA含量分析是诊断肺癌 (尤     

外周血干细胞采集对外周血细胞成分的影响

目的 :探讨连续干细胞采集对外周血细胞成分的影响以及患者对连续采集的耐受性。方法 :在外周血干细胞采集过程中连续检测外周血细胞计数、CD3 4+ 细胞及其亚群 ,造血干细胞培养。结果 :采集后 6例患者出现贫血 ,2 1例原有贫血加重 ,血红蛋白由 (10 7.6± 11.7)g/L降至 (10 0 .3± 11.7)g/L (P <0 .0 1) ;9例出现血小板减少 ,17例原有血小板减少程度加重 ,血小板由采集前的 (96 .46± 5 5 .45 )× 10 9/L降至 (84.71± 14.97)× 10 9/L(P <0 .0 1)。贫血及血小板减少的程度与采集产品中红细胞及血小板的含量显著相关 (P <0 .0 1)。造血干细胞采集后血中的CD3 4+ /CD3 8-细胞以及CD3 4+ /HLA DR-细胞显著增加 ,而BFU E和CFU GM无明显变化。结论 :外周血干细胞采集可使外周血中多能干细胞水平增加 ,但连续采集可使贫血和血小板减少的危险性增加 ,但均可耐受。     

干细胞移植后合并巨细胞病毒感染时淋巴细胞亚群变化

目的　研究异基因外周血干细胞移植 (PBSCT)后早期巨细胞病毒 (CMV)活动性感染时淋巴细胞亚群变化及其意义 ,探讨活动性CMV感染对免疫的影响。方法　根据CMV感染情况 ,将 2 7例早期PBSCT受者分为症状性感染、无症状活动性感染以及同期未出现活动性感染 3组 ,5 1例正常人作为对照组 ,用流式细胞仪测定其淋巴细胞表面CD3 、CD4、CD8、CD16、CD56、CD19、CD2 8的表达 ,比较各组间的差异。结果　PBSCT后早期CD+ 4 T和B细胞数量显著低于正常人 (P值均 <0 0 1) ;2 7例受者中无活动性CMV感染 5例、无症状活动性CMV感染 10例和症状性CMV感染 12例。上述 3组病人的平均CD+ 4 T细胞计数 (× 10 6/L)分别为 32 8± 2 0 3、2 39± 2 18和 199± 92 ;CD+ 8T细胞计数 (× 10 6/L)分别为 4 0 0± 380、2 6 7± 2 0 6和 6 0 3± 4 6 1;CD+ 4 CD+ 2 8的功能细胞亚群的比例分别为 (89 2± 8 9) %、(84 2±10 1) %和 (6 3 5± 11 4 ) % ;自然杀伤 (NK)细胞比例分别为 (16 2± 11 1) %、(2 9 3± 9 9) %和 (19 2±10 2 ) %。与无活动性CMV感染者相比 ,无症状CMV活动性感染者除NK细胞升高外 (P <0 0 1) ,其他指标差异无显著性 ;而症状性CMV感染者和无症状CMV感染者相比 ,其CD+ 4 T细胞数量显著减少(P <0 0 1) ,CD+ 8T     

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.     

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.     

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.     

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.