慢性乙型肝炎树突状细胞表型和功能的变化与免疫耐受

目的 观察慢性乙型肝炎病毒（HBV）感染者树突状细胞（DC）形态和功能的改变。方法 从13例慢性乙型肝炎患者和11例健康人外周血中分离和培养DC,观察DC的形态,用流式细胞仪检测DC表面标记HLA－DR、CD1a、CD80和CD86的表达,用^3H-TdR掺入法测定DC诱导混合性淋巴细胞反应（MLR）的能力。结果 正常DC较慢性乙型肝炎患者在形态上更为典型,不规则,HLA－DR、CD80和CD86分子的表达水平较高（P＜0.05）,诱导MLR的能力较强（P＜0.05）。结论 慢性乙型肝炎患者外周血DC处于不完全成熟状态,其免疫刺激能力较低。     

HBsAg、HBcAg活化树突状细胞治疗慢性乙型肝炎的体外研究

目的 研究慢性乙型肝炎患者外周血树突状细胞(DC)经HBsAg、HBcAg活化后的免疫功能.方法 从慢性乙型肝炎患者外周血中培养扩增DC,在DC成熟前,加入纯的HBsAg、HBcAg刺激,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 经HBcAg刺激DC的CD86表达率为(92.14±5.12)%,明显高于HBsAg刺激组和未加抗原组(P＜0.01);经HBcAg刺激组DC诱导同种异体静止T细胞增殖的能力每分钟液闪计数值(cpm)为34259±3127,明显高于HBsAg刺激组(20258±2917)和单个核细胞组(3469±417),P＜0.01;经HBcAg刺激组DC MLR中IL-12浓度为(342±42.3)ng/L,分别高于HBsAg刺激组和未加抗原组(P＜0.01).结论 体外经HBcAg刺激DC可有效提呈抗原病毒,并可进一步刺激T细胞产生.     

胸腺肽α1对慢性乙型肝炎患者外周血树突状细胞调节作用的体外研究

背景：近年研究显示慢性乙型肝炎（CHB）患者体内树突状细胞（DC）存在不同程度的功能缺失。胸腺肽α1（T-α1）是一种生物学应答调节物质,对CHB的临床治疗有较好疗效。目的：研究T-α1对CHB患者外周血DC分化和功能成熟的影响,寻求改善DC功能的途径。方法：从12例CHB患者和10名健康志愿者的外周血单个核细胞（PBMC）中分离培养DC。培养过程中应用不同浓度T-α1干预DC。以流式细胞仪检测DC表面标志,以甲基噻唑基四唑（MTT）实验检测混合淋巴细胞反应中DC刺激T细胞增殖的能力。结果：从PBMC分离培养得到的外周血DC呈现典型树突状形态。CHB组DC表面标记物CD80、CD86、人白细胞位点DR抗原（HLA-DR）水平以及刺激T细胞增殖的能力显著低于正常对照组（P〈0.001）;经T-α1干预后,两组上述指标均较相应空白对照升高,尤以0.5μg/mlT-α1的作用为著（P〈0.05）,可使CHB组上述指标恢复至正常水平。结论：T-α1能促进CHB患者外周血DC分化和功能成熟,在CHBDC疫苗的研究中可能具有重要价值。     

补肾健脾解毒法中药复方对慢性HBV携带者树突状细胞的影响

目的：研究补肾健脾解毒法中药复方对慢性HBV携带者树突状细胞（DC）的影响。方法：研究分为两个试验组。治疗组纳入30例慢性HBV携带者,对照组纳入10例健康志愿者,分别采用补肾健脾解毒法中药复方、安慰剂干预48周,观察两组人员治疗前后DC的成熟表面标志物、同种混合淋巴细胞反应（MLR）及其细胞因子（IL-12、IFN-γ）的变化。结果：基线时,治疗组患者的DC成熟表面标记物HLA（人白细胞抗原）-DR、CD86、CD80和CD1α的表达水平分别为61.34%、55.78%、42.63%、43.17%,明显低于对照组的90.52%、83.26%、76.81%、84.90%（P〈0.01）;DC诱导MLR的能力、MLR上清液中IL-12和IFN-γ的表达水平全都明显低于对照组（P〈0.01）。治疗48周后,治疗组DC成熟表面标记物的表达水平均较治疗前明显上升（P〈0.01）,但仍显著低于对照组（P〈0.01）;治疗组患者对DC诱导MLR能力也有显著提升,MLR上清液中IL-12和IFN-γ的表达水平明显上升（P〈0.01）,但DC诱导MLR能力及IFN-γ的表达水平未达到对照组水平（P〈0.05或0.01）。结论：补肾健脾解毒法中药复方能促进慢性HBV携带者的DC成熟并改善其免疫功能,从而产生有效的免疫应答以抑制/清除HBV。     

体外负载HSP70-K-ras突变多肽复合物树突细胞的抗胰腺癌作用

背景：K-ras突变多肽难以很好地被树突细胞（DC）捕获,诱导的抗肿瘤效应有限。目的：探讨热休克蛋白70（HSP70）-K-ras突变多肽复合物体外负载于DC的抗胰腺癌作用。方法：使HSP70与K-ras突变多肽在体外结合成复合物,负载于由人外周血单个核细胞体外诱导分化获得的DC。酶联免疫吸附测定（ELISA）检测DC的细胞因子分泌能力,流式细胞仪分析DC负载抗原前后的细胞表型变化,胆囊收缩素（CCK）-8法检测负载抗原后DC对同基因淋巴细胞的促增殖作用,以及激活的同基因淋巴细胞对人胰腺癌细胞株Patu8988、PANC-1和正常人肝细胞株L-02的细胞毒作用。结果：HSP70-K-ras突变多肽复合物可激活DC,最适浓度为0.75μg/ml,可使DC的白细胞介素（IL）-12、肿瘤坏死因子（TNF）-α分泌量和细胞表面CD80、CD83、CD86、HLA-DR表达率显著增高。0.75μg复合物可有效激活1×10^6个DC,刺激1×10^7个同基因淋巴细胞增殖,特异性杀伤具有相同12位点突变类型（GGT→GTT）的Patu8988细胞,杀伤率达52.9%±5.1%,对不同突变类型的PANC-1细胞（GGT→GAT）和正常人肝细胞杀伤作用不明显。结论：负载HSP70-K-ras突变多肽复合物的DC活性增强,可刺激同基因淋巴细胞产生高效而特异的抗胰腺癌效应。     

含CpG基序寡核苷酸对慢性乙型肝炎患者外周血树突状细胞的免疫激活研究

目的 研究含非甲基化CpG基序的寡核苷酸（CpG-ODN）对慢性乙型肝炎患者（CHB）外周血树突状细胞（DC）表型和功能的影响。方法以CD14磁性分选微珠分离CHB患者外周血高纯度单核细胞；以重组人粒细胞巨噬细胞集落刺激因子（hGM—CSF）、重组人白细胞介素-4（hIL-4）诱导扩增DC；以CpG-ODN刺激DC成熟，并与肿瘤坏死因子（TNF）-n比较，评价其对DC表达表面分子人类白细胞组织相容性抗原（HLA）-DR、CD86、CD1a，分泌IL-12p70以及刺激同种T细胞增殖能力的影响。结果与non—ODN和磷酸盐缓冲液（PBS）组比较，CpG-ODN能明显提高CHB患者外周血DC表面分子HLA—DR、CD86的表达，使IL-12分泌增加，刺激同种异体T细胞增殖的能力亦显著增强（P=0．017和0．023），但不能明显提高CD1a的表达；CpG-ODN的上述刺激作用接近或略逊于hTNF-α，但差异无统计学意义（P〉0．05）。结论CpG-ODN与hTNF-α一样能够促进CHB患者外周血DC成熟进而增强其抗原提呈功能。     

冠状动脉支架术后C反应蛋白升高与树突状细胞的关系

目的 :研究冠状动脉支架术后高敏C反应蛋白 (hsCRP)的升高与树突状细胞 (DC)的关系。方法 :术前及术后 2d分离 11例冠状动脉支架术患者外周血单个核细胞 (PBMC) ,在含粒细胞巨噬细胞集落刺激因子(GM CSF)和白细胞介素 4 (IL 4 )的培养条件下制备DC。用流式细胞仪检测DC表面共刺激分子CD86 (B7 2 )的表达 ;混合淋巴细胞反应 (MLR)检测DC对同种异体T淋巴细胞的刺激能力 ;ELISA法测定MLR上清液中的细胞因子 ;比浊法测定hsCRP浓度 ,并探讨CD86表达与hsCRP的相关性。结果 :与术前比较 ,术后 2dDC表面CD86的表达明显增高 ;对T淋巴细胞刺激的能力增强 ;经DC刺激的淋巴细胞分泌促炎细胞因子 (IL 1β、IL 6、TNF α)增多 ,抑炎细胞因子 (IL 10 )减少 ;同时血hsCRP升高 ,且CD86的表达与血hsCRP水平正相关。结论 :支架术后DC的功能被激活 ,由此启动的T淋巴细胞的增殖和炎性反应可能是术后hsCRP升高的原因     

树突状细胞诱导免疫效应细胞对BEL-7402的生长抑制

目的：体内外观察多种细胞因子和（或）肿瘤相关抗原（tumor allogenic antigen,TAA)刺激的正常人树突状细胞（DC）诱导免疫效应细胞对BEL－7402的生长抑制。方法：体外分别用人GM－CSF、IL－4、TNFα、BEL－7402TAA和人IL－2刺激正常人DC和去DC的单个核细胞（免疫效应细胞），5－6d后混合培养DC和免疫效应细胞1－2d。体外实验时效应细胞分无DC刺激组（A0组）、细胞因子培养和DC刺激组（A1组）、细胞因子和TAA培养的DC刺激组（A2组）。体内实验时，裸鼠分为3组：预防组（Ⅱ），于接种BEL－7402前1－2d天给DC激活的免疫效应细胞；治疗组（Ⅲ），待全部裸鼠移植瘤长出时给DC激活的免疫效应细胞；组Ⅱ，Ⅲ于给予DC激活免疫效应细胞后再间断给予DC培养上清液3－6d。对照组（I）给等量的1640培养液。结果：多细胞因子体外可刺激DC发生扩增并高表达B7分子。体外实验中最大杀伤效率：A2组为81％，A1组为68．1％，A0组为3．5％。体内实验中：组I和组Ⅲ第12d时12例全部发生移植瘤；组Ⅱ观察30d时，6例有1例发生移植瘤，观察45d时，仍只有1例发生移植瘤，组I、Ⅱ相差有非常显著意义（P＝0．00466）；在给予DC激活的免疫效应细胞后的第45d处死所有裸鼠并称瘤体的重量，组I、Ⅱ、Ⅲ比较相差有非常显著意义（P＜0．01），组Ⅱ的6只裸鼠仅1只产生移植瘤，瘤重为0．27g。结论：激活的DC在抗恶性肿瘤中有重要作用。     

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.     

依那普利对急性冠脉综合征患者树突状细胞功能的影响

目的 探讨急性冠脉综合征（ACS）患者树突状细胞（DC）功能状态以及依那普利（Enapril）对DC功能的影响.方法 将42例ACS随机分为常规治疗组（21例）和常规治疗加依那普利治疗组（21例）,分别于治疗前及治疗后1个月取血分离外周血T淋巴细胞,用流式细胞仪检测各组DC细胞表面共刺激分子CD86表达（以CD86^＋CD11c^＋细胞比例表示）;混合淋巴细胞反应（MLR）检测DC细胞对正常献血者T淋巴细胞刺激作用;分析依那普利治疗后CD86^＋CD11c^＋细胞比例,MLR的变化及其与血清C-反应蛋白（CRP）的相关性.结果 与正常人比较,ACS患者DC细胞中CD86^＋CD11c^＋细胞比例明显增加（P＜0.001）;对T淋巴细胞增殖刺激作用明显增强（P＜0.001）;与治疗前比较依那普利治疗后CD86^＋CD11c^＋细胞比例和MLR明显下降（P＜0.01）;CD86^＋CD11c^＋细胞比例、MLR均与CRP呈正相关.结论 ACS患者DC细胞功能明显增强;依那普利可能通过抑制DC细胞功能,负向调节患者的特异性免疫而抑制斑块的炎症反应.     

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.     

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.     

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.     

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.