流式检测C1_q~+的细胞表达及在肺结核诊断中的价值

目的初步建立全血细胞中CD+16CD+68CD+14CD-3C1+q的流式检测方法,评价其在结核病诊断中的价值。方法利用流式细胞技术检测CD+16CD+68CD+14CD-3细胞表面C1+q的表达,比较其在健康照(26例),结核潜伏感染者(12例)、活动性结核患者(19例)的表达情况。结果活动性结核患者组外周血CD+14CD-3细胞显著高于健康对照(P0.05);活动性结核患者组CD+16CD+68CD+14CD-3细胞表面C1+q的表达显著高于结核潜伏感染组和健康对照组(P0.05)。结论在活动性结核病人的外周血中,C1+q在CD+16CD+68CD+14CD-3细胞表面高于潜伏感染者,为活动性肺结核患者与潜伏感染者的临床鉴别诊断技术的研究和开发,提供了一条可行的思路。     

结核病患者外周血B细胞及其亚群的检测

目的初步建立B细胞分群技术,并了解健康对照者、结核潜伏感染者、结核患者B细胞亚群的分布情况。方法根据B细胞表面CD19/CD5/CD1d标识将B细胞分为四群。利用流式细胞技术比较健康对照、结核潜伏感染者、结核患者外周血B细胞及B细胞分群表达的差异。结果结核患者外周血B细胞较健康对照者减少,但结果无统计学差异。结核患者外周血CD19+CD5+CD1d+B细胞显著高于健康对照及结核潜伏感染者(P<0.05);CD19+CD5-CD1d+B细胞高于结核潜伏感染者(P<0.05);CD19+CD5-CD1d-B细胞低于结核潜伏感染者(P<0.05)。结论肺结核患者外周血中存在着B细胞亚群的紊乱,尤其是CD19+CD5+CD1d+B细胞增多最为明显,而CD19+CD5-CD1d+B/CD19+CD5-CD1d-B细胞只比结核潜伏感染者显著增多或降低,提示B细胞亚群在结核的发病中起重要作用。     

髓源性抑制细胞在长期抗HBV治疗患者中促进HBsAg阴转的作用和机制探讨

目的研究髓源性抑制细胞(myeloid-derived suppressor cells, MDSCs)、CD4~+T细胞、CD8~+T细胞、P47~(phox)在抗病毒治疗后获得HBsAg清除及未获得HBsAg清除的2种人群外周血中的频率和表达情况,探讨MDSCs在肝炎慢性化及HBsAg清除中的作用及机制。方法筛选2007年7月—2008年7月慢性乙型肝炎(chronic hepatitis B,CHB)患者80例予以抗HBV药物[恩替卡韦(博路定)]治疗。采用流式细胞术检测治疗前CHB组中12例及14例健康对照者外周血MDSCs、CD4~+T细胞、CD8~+T细胞占外周血单个核细胞(peripheral blood mononuclear cells, PBMCs)总数的比例(即MDSCs、CD4~+T细胞、CD8~+T细胞频率)。追踪CHB组患者,排除治疗过程中死亡、失访等患者,72例患者成功随访8年。检测抗病毒治疗8年后8例HBsAg转阴[HBsAg(-)组]与64例未转阴[HBsAg(+)组]患者PBMCs中MDSCs、CD4~+T细胞、CD8~+T细胞频率,用Westernblot技术检测P47~(phox)蛋白的表达。结果治疗前CHB患者PBMCs中MDSCs频率显著高于健康对照者(P 0.05),而PBMCs中CD4~+T细胞、CD8~+T细胞频率与健康对照者无显著差异(P均 0.05)。抗病毒治疗后CHB患者中HBsAg(-)组PBMCs中MDSCs频率低于HBsAg(+)组,CD4~+T细胞及CD8~+T细胞频率显著高于HBsAg(+)组(P均0.05)。抗病毒治疗后HBsAg(+)组外周血中P47~(phox)蛋白表达显著高于HBsAg(-)组(P 0.05)。结论 CHB患者对T细胞耐受可能与MDSCs抑制T细胞应答有关。MDSCs可能通过活性氧途径发挥免疫抑制功能,减少HBsAg清除。     

流式检测CD_(70)~+ CD_(19)~+细胞的表达及其在结核病诊断中的价值

目的初步建立全血细胞中CD+70CD+19的流式检测方法。方法用流式细胞仪检测CD+70CD+19细胞的表达,比较其在肺结核患者(55例)、结核潜伏感染者(27例)及健康对照者(57例)的表达情况。结果结核患者全血中的CD+70CD+19细胞的表达水平明显低于结核潜伏感染组和健康对照组(P0.05)。分别绘制ROC曲线评价CD+70CD+19细胞的诊断效率,在分辨结核病人和健康者时,敏感性92.7%特异性89.5%;在分辨结核病人和潜伏感染者时,敏感性85.5%特异性79.2%。结论 CD+19B细胞CD+70的表达适用于肺结核的诊断及结核潜伏感染者的早期诊断。     

急性髓系白血病患者外周血表达髓系抑制细胞的临床研究

目的:检测急性髓系白血病(AML)患者不同阶段外周血髓系抑制细胞(MDSCs)的表达量,并探讨其与肿瘤相关巨噬细胞(TAM,CD206+)、调节性T细胞(Tregs,CD4+CD25+)的关系。方法:采用流式细胞术检测AML患者30例初诊未治及完全缓解(CR)后外周血表型为CD33+/CD11b+/HLA-DR-/lin-的MDSCs比例,同时检测TAM及Tregs表达水平,并以健康体检者30例作为对照,检测以上指标的变化。结果:1AML患者初诊未治期外周血MDSCs高度表达,显著高于对照者[(5.83±1.66)%∶(0.63±0.31)%,P0.05],经化疗达CR后其数值显著下降,均值为(1.03±0.20)%,CR前、后比较差异有统计学意义(P=0.03);2AML患者初诊未治期外周血表达CD206+的TAM高于对照者[(5.32±1.55)%∶(1.69±0.44)%,P=0.15],经治疗达CR后明显回落;3AML患者初诊未治期外周血表达CD4+CD25+的Tregs明显高于治疗后及对照者[(6.75±0.68)%∶(2.21±0.45)%∶(1.98±0.65)%),P0.01]。结论:MDSCs在AML不同阶段呈规律性表达,与TAM及Tregs表达趋势一致,提示在AML发生、发展过程中MDSCs可能参与了白血病细胞的免疫耐受及肿瘤逃逸机制。     

Tem和Tcm水平变化及对结核患者与潜在感染者的诊断价值

目的探讨效应性记忆T淋巴细胞(effect memory T cells,Tem)和中心型记忆T细胞(center memory T cells,Tcm)表达水平在活动性肺结核与潜伏感染者中的表达差异及其意义。方法选取本院2014年1月-2015年12月确诊的25例活动性肺结核患者(肺结核组)、25例结核分枝杆菌潜伏感染者作为潜伏感染组、30例健康人群作为健康组,采用流式细胞仪技术检测各组外周血CD_4~+记忆T细胞、CD_8~+记忆T细胞的表达水平,并探讨其与高分辨率CT评分(HRCT)的相关性。结果潜伏感染组的CD_4~+Tcm、CD_8~+Tcm水平显著的高于肺结核组、健康组,差异具有统计学意义(P0.05),肺结核组的CD_4~+Tcm、CD_8~+Tcm水平显著的高于肺结核组健康组,差异具有统计学意义(P0.05)。潜伏感染组、肺结核组的CD_4~+Tem、CD_8~+Tem水平显著的低于健康组,差异具有统计学意义(P0.05),肺结核组、潜伏感染组的CD_4~+Tem、CD_8~+Tem水平差异无统计学意义(P0.05)。肺结核患者CD_4~+Tcm、CD_8~+Tcm水平与HRCT评分呈显著的负相关关系(P0.05);肺结核患者CD_4~+Tem、CD_8~+Tem水平与HRCT评分无显著的相关系(P0.05)。结论 CD_4~+、CD_8~+中心记忆性T细胞在活动性肺结核与潜伏感染者中水平差异显著,并且与肺结核患者病例损伤程度有关。     

人类免疫缺陷病毒合并结核分枝杆菌感染者的抗结核细胞免疫功能

目的 评价HIV合并结核分枝杆菌潜伏感染或合并活动性结核患者的抗结核细胞免疫功能.方法 应用早期分泌性抗原靶蛋白(ESAT)-6和培养滤出蛋白(CFP)-10诱导的结核酶联免疫斑点法对云南地区100例明确诊断的HIV感染者的血液标本进行结核分枝杆菌特异性T淋巴细胞检测,同时应用流式细胞仪检测外周血CD3+CD4+T淋巴细胞和CD3+CD8+T淋巴细胞的绝对计数水平.采用Mann-Whitney检验进行非参数统计分析.结果临床上无活动性结核感染证据的HIV感染者中合并结核分枝杆菌潜伏感染的感染率高达67.6%.HIV合并结核分枝杆菌潜伏感染者的外周血CD3+CD4+T淋巴细胞(532×106/L)和CD3+CD8+T淋巴细胞(473×106/L)绝对计数与单纯HIV感染者(406×106/L和504 × 106/L)相比,差异无统计学意义.HIV合并活动性结核感染者的外周血CD3+CD4+T淋巴细胞绝对计数平均值为189 × 106/L,CD3+CD8+T淋巴细胞绝对计数平均值为293×106/L,均显著低于单纯HIV合并结核潜伏感染组和HIV组(U=168.0,U=163.0;U=147.0,U=374.0;均P<0.01).HIV合并活动性结核感染者的ESAT-6和CFP-10抗原特异性斑点形成细胞数(31/106细胞和82/106细胞)显著低于HIV合并结核分枝杆菌潜伏感染者(92/106细胞和109/106细胞.U=507.0,U=529.5,均P<0.01).结论 我国无活动性结核临床证据的HIV感染人群中有较高的结核潜伏感染率,HIV合并活动性结核感染者的总体细胞免疫应答功能及特异性抗结核免疫应答功能均严重受损.     

强直性脊柱炎患者外周血CD4+CD25+调节性T细胞的研究

目的 观察强直性脊柱炎(AS)患者外周血CD4+CD25调节性T细胞(Treg)的数量、功能及肿瘤坏死因子(TNF)-a 抑制剂治疗的影响.方法 活动性AS患者25例,10例给予etanercept治疗12周,健康对照21名,分离外周血单个核细胞(PBMC),流式细胞术检测CD4+CD25high T细胞比例;实时定量聚合酶链反应检测FOXP3 mRNA表达;免疫磁珠法去除CD25+细胞,可溶性噻唑盐(WST-1)法检测T细胞增殖.结果 活动性AS组CD4+CD25high T细胞比例与对照组差异尤统计学意义,但FOXP3 mRNA表达明显低于对照组(P＜0.01),并与C反应蛋白(CRP)呈负相关(P＜0.01).两组的CD4+CD25+细胞体外均能抑制T细胞增殖(P均＜0.01). Etanercept治疗明显增加CD4+CD25highT细胞比例和FOXP3 mRNA表达(P均＜0.01),与CRP降低呈负相关(P＜0.05;P＜0.01).结论 AS患者外周血表达FOXP3的CD4+CD25+Treg细胞异常,可能参与AS发生和发展;Etanercept治疗上调Treg细胞,可能是抗TNF-α治疗的一个机制.     

T细胞、NKT和NK细胞在结核病诊断中的意义探讨

目的探讨CD3+T细胞、CD3+CD16+CD56+N KT细胞及CD3-CD16+CD56+N K细胞在肺结核患者外周血的表达状态及其临床意义。方法采用流式细胞仪分析123例肺结核患者(73例初治肺结核和50例复治肺结核)及83例健康者(43例结核菌素皮试阳性和40例结核菌素皮试阴性)外周血中T细胞、NKT和NK细胞的表达率。结果初、复治肺结核患者的NKT细胞表达率明显高于健康者(P0.01),但初治肺结核患者的NK细胞、T细胞的表达率与健康者相比差异无统计学意义(P0.05)。T细胞表达率在不同程度的初治肺结核患者中差异有统计学意义(P0.01),疾病越重,表达越高。NKT、NK细胞差异无统计学意义(P0.05)。复治肺结核患者的T细胞表达率显著高于初治肺结核患者(P0.01),但NK细胞、NKT细胞的表达率在2组间差异无统计学意义(P0.05)。结论T细胞、NKT细胞是结核病细胞免疫的重要组成部分。对NKT细胞只在活动性肺结核患者才会呈现高表达,测定外周血NKT细胞可能成为辅助诊断活动性肺结核的手段之一。     

结核患者的CD_(39)~+调节性T细胞的免疫研究

目的研究外周血中CD+4CD+25CDlow127细胞与CD+39CD+4CD+25CDlow127细胞在健康人、隐性结核感染者和活动性结核患者中表达水平的差异。方法检测10例健康对照者、26例隐性感染结核者及35例活动性结核感染者外周血的CD+4CD+25CDlow127细胞与CD+39CD+4CD+25CDlow127细胞的表达水平。结果外周血的CD+4CD+25CDlow127细胞在健康对照组、隐性感染组及活动性结核组的表达分别为(4.84±0.85)%、(6.95±0.64)%、(9.49±1.14)%。三组之间比较有显著性差异(P均0.01)。外周血的CD+39CD+4CD+25CDlow127细胞在健康对照组、隐性感染组及活动性结核组的表达分别为(3.24±0.60)%、(4.88±0.54)%、(7.48±0.83)%,组别之间比较有显著性差异(P均0.01)。结论 CD+4CD+25CDlow127细胞与CD+39CD+4CD+25CDlow127细胞在活动性结核与隐匿性结核中的表达上差异或与免疫抑制和免疫耐受的强度有关,是区分两者的优选指标。     

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.