T淋巴细胞活化诱导的细胞死亡在肾综合征出血热发病中的作用

目的研究肾综合征出血热(HFRS)患者外周血T淋巴细胞(PBL-T)活化诱导细胞死亡(activation-inducedcelldeath,AICD)的发生情况,探讨AICD在HFRS发病机制中的作用。方法以免疫磁珠负性分离PBL-T;应用植物血凝素(PHA)和离子霉素(ionomycin)体外培养诱导细胞活化;采用HE染色和脱氧核苷酸原位末端标记法(TUNEL)染色观察凋亡情况并用流式细胞仪检测PBL-T的凋亡率。应用Westernblot检测HFRS患者外周血T淋巴细胞caspase-3的表达情况,用RT-PCR对汉坦病毒的特异性基因片段进行检测。结果HFRS患者诱导组PBL-T凋亡率为(27.79±0.99)%,明显高于未诱导组的(17.16±1.14)%(P<0.01);无论是否诱导,患者PBL-T的凋亡率均高于健康人。HFRS患者诱导组PBL-Tcaspase-3的17000活性成分表达显著增加。扩增产物测序结果,有19例患者血清出现阳性条带,其中汉滩病毒7例,汉城病毒12例。结论以PHA和离子霉素可有效地诱导体外培养的PBL-T发生AICD。HFRS患者PBL-T经激活诱导后凋亡率明显提高。有79.2%的患者可以扩增到汉坦病毒特异性的基因片段。HFRS患者PBL-T凋亡相关蛋白caspase-3的表达增加,提示AICD可能与HFRS的发病机制有关。     

慢性乙型肝炎患者外周血淋巴细胞凋亡与HBVDNA及Th1/Th2类细胞因子的关系

研究活化诱导的外周血单核细胞凋亡 (AICD)在乙型肝炎慢性化中的作用以及与HBVDNA和Th1/Th2的关系。分离 2 5例慢性乙型肝炎患者与 10例健康人外周血单个核细胞 (PBMC) ,在PHA的刺激下培养 ,经流式细胞仪检测凋亡 ,ELISA检测细胞因子IFN -γ、IL - 4、IL - 10。经过PHA诱导的PBMC凋亡率乙型肝炎组明显高于正常对照组 (P <0 0 1)。在慢乙肝组中 ,随着HBVDNA的上升 ,凋亡比率升高 ,IL - 4与凋亡成正相关 ,有统计学差异(P <0 0 0 1)。AICD可能在乙型肝炎感染慢性化与形成免疫耐受过程中起到重要作用 ,HBVDNA与AICD有关 ,IL -4促进AICD的发生     

慢性乙型肝炎患者外周血树突状细胞诱导特异性T淋巴细胞应答

目的 探讨慢性乙型肝炎患者外周血树突状细胞(Dc)是否诱导特异性T细胞应答。方法(1)将研究对象分为慢性乙型肝炎患者组、急性乙型肝炎痊愈组、健康志愿者组,分离各组研究对象的外周血单个核细胞(PBMC),细胞内细胞因子染色方法检测其对细胞毒性T淋巴细胞(CTL)特异表位多肽乙型肝炎病毒核心抗原(HBcAg)18-27的记忆性免疫应答;(2)培养慢性乙型肝炎患者DC,将负载有乙型肝炎抗原表位多肽的DC诱导特异的T细胞应答。采用细胞内细胞因子染色方法检测诱导的T细胞分泌的细胞因子,乳酸脱氢酶释放法测定诱导的T细胞杀伤活性。结果(1)急性乙型肝炎患者PBMC对HBcAg 18-27 CTL特异表位多肽存在记忆的免疫应答,其分泌干扰素-γ的CD8+T细胞占CD8+T细胞总数的(4.3±2.5)％,分泌白细胞介素-2的占总细胞数的(4.8±2.2)％,分泌肿瘤坏死因子-α占总细胞数的(4.6±2.3)％。而慢性乙型肝炎患者和健康志愿者对其记忆应答很低,与急性乙型肝炎患者比较差异有显著性,t值为2.508-3.305,P<0.05。(2)用多肽共孵育过的慢性乙型肝炎患者DC多次诱导的T细胞慢性乙型肝炎患者组,加肽孵育的靶细胞比例为30:1、10:1、3:1时,其杀伤率分别为(57.0±20.3)％、(49.5±20.2)％、(21.8±12.9)％,均高于对照组,表明慢性乙型肝炎患者DC可以诱导特异的T     

慢性乙型肝炎患者外周血树突状细胞功能的研究

目的研究慢性乙型肝炎患者外周血树突状细胞(DC)的免疫功能.方法从慢性乙型肝炎患者外周血中分离单个核细胞,用无血清培养法分离培养DC,用流式细胞仪检测DC表型,用液闪计数仪观察DC对T细胞的增殖作用,用ELISA法检测混合淋巴细胞反应(MLR)中细胞因子的分泌水平.结果患者组DC的CD86的表达率为(70.2±5.2)%,明显低于正常人组(95.3±3.8)%,P＜0.01;其诱导T细胞增殖能力每分钟液闪计数cpm为10000±2000,明显低于正常人组(cpm为30000±3000),P＜0.01患者MLR中IL-12为(120.0±19.7)pg/ml,γ-干扰素为(799.0±161.3)pg/m1,明显低于正常人组的(280.0±41.1)pg/ml和(3359.0±635.4)pg/ml,P＜0.01.结论慢性乙型肝炎患者外周血DC免疫功能低下,并与DC表面CD86的表达率下降及DC分泌IL-12减少密切相关.     

急性白血病化疗24小时体内细胞凋亡预测化疗疗效29例

目的 :观察体内细胞凋亡和急性白血病化疗的关系 ,为白血病个体化治疗提供早期预测疗效的指标。方法 :采用流式细胞术膜联蛋白 (AnnexinV)法检测 2 9例急性白血病患者化疗前及化疗 2 4h体内细胞凋亡 ,并观察其与患者化疗一疗程疗效之间的关系。结果 :化疗前和化疗 2 4h急性白血病细胞的凋亡率分别为(5 .74± 2 .4 7) %和 (9.77± 3.5 4 ) % ,化疗后凋亡细胞显著增加 (P <0 .0 1)。将完全缓解及部分缓解患者归为有效组 ,未缓解患者归为无效组。有效组化疗 2 4h的细胞凋亡率为 (11.4 5± 3.0 9) % ,细胞凋亡增加 (5 .91±3.4 8) % ;无效组化疗 2 4h的细胞凋亡率为 (8.15± 3.6 6 ) % ,细胞凋亡增加 (2 .2 8± 3.5 8) %。有效组化疗后凋亡增加明显高于无效组 (P <0 .0 5 )。其特异性为 83.3% ,敏感性为 84 .6 %。结论 :AnnexinV法检测白血病细胞体内凋亡是一个较为敏感的预测化疗疗效的早期指标     

激活诱导细胞凋亡(AICD)在乙型肝炎慢性化机制中的意义

探讨激活诱导细胞凋亡(AICD)在乙型肝炎病毒(HBV)慢性感染中的意义。通过流式细胞仪检测慢性乙型肝炎(CHB)患者外周血淋巴细胞(PBL)在PHA激活前后的凋亡率,采用实时荧光定量PCR对血清HBVDNA进行定量检测。按HBVDNA含量(拷贝/ml)把CHB患者分为高病毒载量组(≥107/ml)、中载量组(105~106/ml)和低载量组(<105/ml)。静息淋巴细胞凋亡率在CHB患者组与正常对照组都比较低,两组比较差异无显著性(P>0.05);但与正常对照组比较,AICD在CHB患者组明显升高,而在重型肝炎组则降低,有显著性差异;AICD在高病毒载量组最高,低病毒载量组最低。研究提示AICD可能参与CHB的慢性化及重型化并随病毒载量的增高而加重。     

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。     

毫米波诱导人肝癌细胞株凋亡的研究

目的　研究 35 .8GHz毫米波照射诱导人肝癌细胞株BEL74 0 4凋亡。方法　选取BEL74 0 4人肝癌细胞体外培养 ,随机分为 4组 ,分别为对照组、毫米波照射 30min组、氟尿嘧啶 (5 FU)组、毫米波联合 5 FU组。用荧光显微镜观察细胞核的变化 ;DNA片段化分析检测染色体断裂情况 ;用流式细胞仪检测凋亡率。结果　BEL74 0 4细胞经毫米波照射后诱发的细胞死亡具有细胞凋亡的形态学特征 ;照射 30min组和 5 FU组诱导细胞凋亡的比例高于对照组 ,分别为 (14 .33± 2 .6 6 ) % ,(18.5 8±2 .5 7) %和 (3.2 1± 1.0 6 ) % ,毫米波与 5 FU联合明显增加细胞凋亡率 ,达 (2 7.91± 3.6 6 ) %。结论　 35 .8GHz毫米波照射可诱导BEL74 0 4肝癌细胞凋亡 ,与 5 FU联合使用可明显增加凋亡比例     

慢性阻塞性肺病患者外周血淋巴细胞激活诱导凋亡的检测

激活诱导细胞死亡(activationinduced cell death,AICD)也称细胞凋亡,外周血淋巴细胞(peripheral blood lympho-cytes,PBL)在抗原、丝裂原等刺激下可引起AICD。为了探讨慢性阻塞性肺病(chronic obstructive pul monary disease,COPD)患者PBL的AICD水平,作者采用丝裂原PHA-P分别与正常人及COPD患者PBL体外培养,通过流式细胞仪检测PBL凋亡率,了解COPD患者PBL凋亡变化,探讨COPD患者的免疫损伤机制。1资料与方法选择吉林大学第四医院呼吸科住院COPD患者共39例,男22例.女17例,年龄60~80岁,均符合中华医学会呼吸病学分会慢性…     

多巴胺对体外培养神经细胞的影响

目的　研究高浓度多巴胺 (DA)对神经细胞的毒性作用。　方法　观察不同浓度、不同时间DA对体外培养神经细胞的毒性 ,并用琼脂糖凝胶电泳、原位末端标记、流式细胞仪进行细胞凋亡的检测。　结果　 (1)高浓度DA引起神经细胞凋亡 ;(2 )给予 80、16 0、32 0 μmol/LDA处理 2 4h后神经细胞凋亡率分别为 (41 49± 1 32 ) %、(6 5 6 7± 1 6 8) %、(84 6 5± 2 2 1) % ,明显高于浓度效应对照组〔(2 0 7± 0 2 6 ) % ,P <0 0 5〕 ;(3)在给予 30 0 μmol/LDA处理的条件下 ,经过 12、2 4、48h后 ,神经细胞的凋亡率分别为 (37 90± 0 39) %、(6 9 34± 3 31) %、(82 0 7± 0 86 ) % ,明显高于时间效应对照组〔(2 30± 0 11) % ,P <0 0 5〕。　结论　高浓度DA对体外培养神经细胞具有细胞毒性 ,诱导神经细胞凋亡 ,具有时间及剂量效应。     

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.     

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.     

N-acetyl-L-cysteine combined with mesalamine in the treatment of ulcerative colitis： Randomized,placebo-controlled pilot study

AIM： To evaluate the effectiveness and safety of oral N-acetyl-L-cysteine （NAC） co-administration with mesalamine in ulcerative colitis （UC） patients.
METHODS： Thirty seven patients with mild to moderate UC were randomized to receive a four-wk course of oral mesalamine （2.4 g/d） plus N-acetyl-L-cysteine （0.8 g/d） （group A） or mesalamine plus placebo （group B）. Patients were monitored using the Modified Truelove-Witts Severity Index （MTWSI）. The primary endpoint was clinical remission （MTWSI ≤ 2） at 4 wk. Secondary endpoints were clinical response （defined as a reduction from baseline in the MTWSI of ≥ 2 points） and drug safety. The serum TNF-α, interleukin-6, interleukin-8 and MCP-1 were evaluated at baseline and at 4 wk of treatment. RESULTS： Analysis per-protocol criteria showed clinical remission rates of 63% and 50% after 4 wk treatment with mesalamine plus N-acetyl-L-cysteine （group A） and mesalamine plus placebo （group B） respectively （OR = 1.71; 95% CI： 0.46 to 6.36; P = 0.19; NNT = 7.7）. Analysis of variance （ANOVA） of data indicated a significant reduction of MTWSI in group A （P = 0.046） with respect to basal condition without significant changes in the group B （P = 0.735） during treatment. Clinical responses were 66% （group A） vs 44% （group B） after 4 wk of treatment （OR = 2.5; 95% CI： 0.64 to 9.65; P = 0.11; NNT = 4.5）. Clinical improvement in group A correlated with a decrease of IL-8 and MCP-1. Rates of adverse events did not differ significantly between both groups.
CONCLUSION： In group A （oral NAC combined with mesalamine） contrarily to group B （mesalamine alone）, the clinical improvement correlates with a decrease of chemokines such as MCP-1 and IL-8. NAC addition not produced any side effects.     

Delorme's transrectal excision for internal rectal prolapse

Surgical therapy of functional outlet obstruction in patients with internal rectal intussusception may include abdominal, perineal, or transrectal procedures. Because abdominal procedures often result in significant physiologic impact but unrelieved constipation, the authors have elected Delorme's transrectal excision for management of these patients. Since a short-term placebo effect attends many therapies, this report describes results of transrectal excision only after a threeyear postoperative period. Delorme's transrectal excision of internal intussusception accomplished sustained symptomatic relief in over 70 percent of otherwise refractory constipated patients. The association of internal intussusception with other abnormalities underscores the importance of defining both anatomic and functional components when selecting patients whose constipation may require surgical therapy. Critical technical elements, surgical pitfalls, and potential complications of the procedure are discussed.Poster presentation at the meeting of The American Society of Colon and Rectal Surgeons, Toronto, Canada, June 11 to 16, 1989.     

The oral glucose tolerance test: A comparison of the time points on the basis of limit values,normal dispersion,and reproducibility

Summary Time points in the glucose tolerance test (GTT) are compared on the basis of limit values, dispersion within a reference population, and reproducibility. We suggest using the distance between a limit value and the median reference value as a measure of the magnitude of abnormality. The distance between 140 mg/100 ml and the median fasting plasma glucose value is chosen as a standard distance and limits for other points in the GTT are calculated to equal this standard distance of abnormality. We suggest that the probability of correctly interpreting an inividual result is directly related to the reproducibility of the test and inversely related to the percentage of the total range of values which is dispersed among the normal population. The ratio of reproducibility to percentage normal dispersion is proposed as an index of the probability of correctly interpreting an individual result. According to this index, the probability of correct interpretation varies in order: fasting plasma glucose concentration>3-h>2-h>0.5-h>1-h plasma glucose concentration.