特发性膜性肾病患者调节性T细胞和B淋巴细胞的变化及其临床意义

目的:观察特发性膜性肾病(IMN)患者外周血调节性T细胞、B淋巴细胞亚群以及肾组织中B淋巴细胞浸润分布的变化,探讨与膜性肾病(MN)发病机制及临床表现的关系. 方法:66例(男性45例,女性21例,年龄15～71岁)经肾活检确诊为MN,排除各种继发因素,近期未使用大剂量免疫抑制剂的IMN患者,流式细胞仪测定外周血调节性T细胞(CD4+CD25+Foxp3+Treg细胞)、B淋巴细胞(CD20+细胞)和T淋巴细胞亚群(CD3+、CD4+、CD8+细胞)的计数;另采用免疫组织化学方法检测患者肾组织中B淋巴细胞数量及分布.40例年龄、性别匹配的健康志愿者作为对照.根据患者尿蛋白水平,将其分为大量蛋白尿组(尿蛋白定量≥3.5 g/d)及非大量蛋白尿组(尿蛋白定量＜3.5g/d).比较IMN患者外周血调节性T细胞,B淋巴细胞及T淋巴细胞亚群与正常对照的差别,以及IMN患者中大量蛋白尿者和非大量蛋白尿者之间上述淋巴细胞亚群水平的差异及其可能的临床意义. 结果:(1)IMN患者外周血调节性T细胞计数较正常对照组显著降低[(23.2±10.7) cells/μl vs (31.4±7.29) cells/μl,P<0.01].(2)与正常对照组比较,IMN患者外周血B淋巴细胞计数显著升高[(208±111) cells/μl vs (137±64) cells/μl,P<0.01].(3)IMN患者和正常对照组相比,其CD4+细胞计数有升高趋势[(715±267) cells/μl vs (657±214) cells/μl,P>0.05],CD8+细胞计数有降低趋势[(393±178) cells/μl vs (464±200) cells/μl,P>0.05],而CD4+/CD8+比值较正常对照显著升高(2.04±0.92 vs 1.52±0.48,P<0.01).(4)大量蛋白尿组B淋巴细胞计数明显高于非大量蛋白尿组[(246±117) cells/μl vs (186±108) cells/μl,P<0.05],但两组之间调节性T细胞、CD4+细胞、CD8+细胞计数及CD4+/CD8+比值均无统计学差异(P>0.05).(5)IMN患者肾组织中B淋巴细胞浸润数量较正常对照升高,伴肾组织B淋巴细胞灶性聚集者其尿NAG酶水平升高. 结论:IMN患者表现出调节性T细胞水平降低,B淋巴细胞水平上调和CD4+/CD8+细胞比值的偏移.外周血B淋巴细胞水平升高与IMN患者大量蛋白尿关系密切,肾组织中B淋巴细胞浸润数目较正常对照升高,存在局灶B淋巴细胞聚集的患者小管间质急性损伤加重.对IMN患者B淋巴细胞的数目和组织分布进行观察,有可能为监测病情、指导治疗提供帮助.     

Ⅳ型和Ⅴ型狼疮性肾炎患者外周血与肾组织淋巴细胞亚群的变化和临床意义

目的:观察Ⅳ型与Ⅴ型狼疮性肾炎(LN)患者外周血淋巴细胞亚群的变化和肾组织淋巴细胞亚群和单核-巨噬细胞的浸润情况,并进一步分析与临床和病理的关系.方法:52例(男性11例,女性41例,年龄18～55岁)初治活动性系统性红斑狼疮,根据2003年ISN/RPS病理分型标准经肾活检确诊为Ⅳ型(n=32)和Ⅴ型(n=20),流式细胞仪测定外周血淋巴细胞亚群(CD3 、CD4 、CD8 、CD20 细胞)和CD4 CD25 Foxp 3 调节性T细胞(Treg细胞)的比例与计数,计算CD4 /CD8 比值.20例年龄、性别匹配的健康志愿者作为对照.同时检测30例Ⅳ型和11例Ⅴ型LN肾组织CD4、CD8、CD20和CD68免疫组化染色,取10例移植肾供肾活检标本作为正常肾组织对照.从中再选取20例(Ⅳ型和Ⅴ型各10例)行CD4和Foxp 3双标免疫组化染色.比较LN患者外周血淋巴细胞亚群和Treg细胞与正常对照的差别,分析Ⅳ型和Ⅴ型之间外周血和淋巴细胞亚群和Treg细胞的变化特点.进一步分析Ⅳ型和Ⅴ型LN肾组织淋巴细胞亚群,特别是Treg以及单核-巨噬细胞浸润的差异和临床病理联系.结果:活动性Ⅳ型、Ⅴ型LN患者与正常人比较外周血CD4 细胞比例、计数和CD4 /CD8 细胞比值及Treg细胞显著降低,CD8 细胞及CD20 细胞明显升高(P＜0.01).Ⅳ型与Ⅴ型相比:(1)外周血CD4 细胞比例[(25.9±6.86)％vs(31.0±7.59)％,P＜0.05]、计数[(288±173)个/μlvs(420±165)个/μl,P＜0.01]和CD4 /CD8 比例降低更明显(0.74±0.31vs1.06±0.57,P＜0.05);(2)外周血CD20 细胞比例无明显差异,但Ⅴ型CD20 细胞计数升高较Ⅳ型明显[(185±136)个/μlvs(268±179)个/μl,P＜0.05];(3)Treg细胞比例[(0.82±0.4)％vs(1.31±0.7)％,P＜0.05]和计数[(8.19±4.26)个 /μlvs(17.5±10.0)个/μl,P＜0.01]Ⅳ型降低更明显.CD4 细胞比例和计数与AI呈负相关(r分别为-0.281和-0.380,P＜0.05),CD4 /CD8 细胞比值与SLEDAI呈负相关(r=-0.307,P＜0.05).Treg细胞比例和计数与SLEDAI呈显著负相关,相关系数分别为-0.411(P＜0.01)和-0.480(P＜0.01),与肾组织AI呈负相关(r为-0.325和-0.473,P＜0.01).肾组织免疫组化见Ⅳ型肾间质浸润的CD4 、CD8 、CD20 和CD68 细胞均高于Ⅴ型病例,其中肾小管浸润的CD20 细胞Ⅳ型明显高于Ⅴ型(119.3±89.7vs36.0±36.0,P＜0.01),肾小球内浸润的CD68 细胞显著高于Ⅴ型(12.4±8.8vs1.7±2.31,P＜0.01).正常肾组织不表达Foxp 3 Treg细胞,绝大多数Foxp 3表达于CD4 细胞,LN中CD4 Foxp 3 Treg细胞主要位于肾间质,Ⅳ型LN CD4 Foxp 3 Treg细胞明显高于Ⅴ型(27.6±18.0 vs2.8±5.0,P＜0.01).结论:Ⅳ型LN患者外周血CD4 细胞数量、CD4 /CD8 细胞比例和Treg细胞数量较Ⅴ型明显降低,Ⅳ型LN肾组织T、B淋巴细胞,特别是调节性T细胞和巨噬细胞浸润较V型明显升高,提示不同病理类型LN的免疫发病机制不同.     

皮肌炎与多发性肌炎临床表现和实验室检查对比分析

目的比较皮肌炎(DM)和多发性肌炎(PM)临床表现和实验室检查的异同。方法回顾性分析2012年1月1日至2015年10月31日中国医科大学附属第一医院风湿免疫科收治的62例DM和29例PM患者的临床资料,分析其临床特征的异同。并比较伴有间质性肺病(ILD)的DM和PM患者实验室检查的异同。结果乏力、肌痛、吞咽困难、发热等症状的发生频率在DM和PM组间差异均无统计学意义。DM组外周血淋巴细胞计数为(1.13±0.48)×10~9/L,显著少于PM组的(1.78±0.90)×10~9/L(P=0.01)。DM患者CD3~+T细胞数[(612±332)个/μL对(1543±945)个/μL,P=0.01]、CD4~+T细胞数[(370±175)个/μL对(741±450)个/μL,P=0.02]、CD8+T细胞数[(219±178)个/μL对(738±644)/μL,P=0.02]均显著少于PM组。PM患者CD4+T/CD8+T比值为(2.6±1.5),显著高于DM患者的(1.5±1.0)(P=0.03)。与PM患者相比,DM患者血清白蛋白水平[(34.2±4.7)g/L对(38.1±5.5)g/L,P=0.01]明显降低。伴有ILD的DM(DM-ILD)患者外周血淋巴细胞计数[(1.47±1.04)×10~9/L对(1.70±0.85)×10~9/L,P=0.04]、血清肌酸激酶水平显著低于伴有ILD的PM患者(PM-ILD)[(1220.8±2118.5)U/L对(3150.8±2965.9)U/L,P=0.02]。DM-ILD患者CD4~+T/CD8~+T比值为2.3±1.4,显著高于PM-ILD患者的1.4±0.4(P=0.00)。结论 DM和PM在临床症状上具有一定相似点,但外周血T细胞总数及不同亚群计数却存在显著差异,提示免疫失衡在DM患者的发病机制中具有重要作用。     

Notch信号通路对慢性丙型肝炎患者CD8~+T淋巴细胞功能的影响

目的观察Notch信号通路对慢性丙型肝炎患者CD8~+T淋巴细胞功能的影响,初步阐释Notch信号通路在慢性丙型肝炎发病中的作用。方法收集2016年1月-8月在陕西省人民医院就诊的慢性丙型肝炎初治患者38例和健康志愿者17例,分选CD8~+T淋巴细胞,应用实时定量PCR法检测CD8~+T淋巴细胞中Notch1和Notch2 mRNA的表达水平。分别应用直接接触培养系统和间接接触培养系统将从HLA-A2限制性丙型肝炎患者分选的CD8~+T淋巴细胞与HCV感染的Huh7.5细胞共培养,加入Notch信号通路抑制剂DAPT,通过检测乳酸脱氢酶和细胞因子水平分析CD8~+T淋巴细胞的细胞毒性和非细胞毒性功能的变化。2组间计量资料的比较采用t检验。结果 Notch1和Notch2 mRNA在慢性丙型肝炎患者CD8~+T淋巴细胞中的表达水平较健康志愿者均显著升高,分别升高约10倍和4倍,差异均有统计学意义(t值分别为6.63、18.88,P值分别为0.003、0.001)。抑制Notch信号通路可增加直接接触培养系统中细胞死亡的比例[(26.41±4.76)%vs(13.65±4.65)%,t=3.32,P=0.029];但抑制Notch信号通路对间接接触培养细胞中CD8~+T淋巴细胞毒性无明显影响。抑制Notch信号通路后,IFNγ的表达在直接接触培养系统[(52.19±12.38)pg/ml vs(18.62±5.66)pg/ml,t=4.27,P=0.013]和间接接触培养系统[(18.86±3.09)pg/ml vs(5.52±2.52)pg/ml,t=5.79,P=0.004]中均显著升高。TNFα的表达在抑制Notch信号通路后仅在直接接触培养系统中显著升高[(1296.0±293.3)pg/ml vs(585.3±32.62)pg/ml,t=4.17,P=0.014],而在间接接触培养系统中无明显变化[(297.3±175.4)pg/ml vs(273.7±105.9)pg/ml,t=0.20,P=0.851]。结论抑制Notch信号通路可增强慢性丙型肝炎患者中CD8~+T淋巴细胞的细胞杀伤和非细胞杀伤功能,Notch信号通路在慢性HCV感染中可能通过抑制CD8~+T淋巴细胞功能发挥诱导免疫耐受的作用。     

胸腺肽α1对慢性乙型肝炎CD4~+CD25~+CD127~(dim/-)调节性T淋巴细胞功能的影响

目的观察胸腺肽α1对慢性乙型肝炎CD4~+CD25~+CD127~(dim/-)调节性T淋巴细胞(Treg)功能的影响。方法选取2016年12月-2017年7月陕西省人民医院感染性疾病科收治的慢性乙型肝炎患者67例,其中38例接受恩替卡韦抗病毒治疗(对照组),29例接受恩替卡韦联合注射用胸腺肽α1治疗(治疗组),分别于治疗前后分离外周血单个核细胞,流式细胞术检测CD4~+CD25~+CD127~(dim/-)Treg水平,纯化CD4~+CD25~+CD127~(dim/-)Treg,与自体CD4+CD25-T淋巴细胞共培养,CCK-8法检测细胞增殖,ELISA法检测细胞因子分泌水平。计量资料2组间比较采用t检验,计数资料组间比较采用χ2检验。结果对照组、治疗组患者经治12周CD4~+CD25~+CD127~(dim/-)Treg均较基线水平明显下降,差异均有统计学意义(8.85±2.18)%vs(12.32±1.22)%、(9.26±2.30)%vs(13.71±2.32)%,t值分别为4.579、4.803,P值分别为0.0005、0.0003]。选取17例对照组患者和21例治疗组患者检测细胞因子分泌水平。经治12周对照组患者治疗12周后Treg与CD4+CD25-T淋巴细胞共培养系统中细胞增殖和分泌细胞因子水平均无明显变化;治疗组共培养系统中细胞计数较基线水平明显升高,差异有统计学意义[(3.66±0.95)×106个vs(2.07±0.51)×106个,t=5.709,P0.0001];治疗组共培养细胞分泌抑制性细胞因子IL-10、IL-35水平明显降低,差异均有统计学意义[(41.40±11.89)pg/ml vs(56.53±27.85)pg/ml、(122.9±9.98)pg/ml vs(130.0±15.98)pg/ml,t值分别为2.639、2.459,P值分别为0.019、0.028)],分泌抗病毒细胞因子IFNα、IFNγ水平明显升高,差异均有统计学意义[(297.5±83.56)pg/ml vs(235.6±67.72)pg/ml、(5.83±0.85)pg/ml vs(4.39±0.95)pg/ml,t值分别为2.603、4.659,P值分别0.017、0.0004]。结论胸腺肽α1能够抑制慢性乙型肝炎CD4~+CD25~+CD127~(dim/-)Treg水平及免疫抑制功能,提高患者免疫功能。     

硒对氟中毒鸡淋巴细胞亚群及脾淋巴细胞凋亡的影响

目的 观察硒对慢性氟中毒鸡血液、脾脏CD4~+,CD8~+淋巴细胞亚群以及脾淋巴细胞凋亡的影响,并探讨其作用机制.方法 将8日龄海蓝褐雏鸡180只按体质量随机分为3组:对照组、氟中毒组、硒拮抗组,每组60只,分别饲以含氟195、1000、1000 ms/kg、含硒0.08、0.08、4.00 mg/kg的全价日粮.于30、60、90 d用流式细胞术检测外周血和脾脏中CD4~+、CD8~+淋巴细胞水平,TUNEL法检测脾淋巴细胞凋亡情况.结果 与对照组比较,30、60、90 d氟中毒组外周血CD4~+淋巴细胞水平降低[(35.36±4.27)%vs(24.29±2.96)%、(47.65±5.42)%vs(41.62±3.96)%、(49.58±3.98)%vs(42.35±6.03)%,P<0.05或<0.01],CD4~+/CD8~+比值也明显降低[(1.701±0.145)%vs(1.393±0.163)%、(2.712±0.345)%vs(1.781±0.201)%、(2.438±0.356)%vs(1.973 ±0.229)%,P<0.05或<0.01];与氟中毒组比较.30、60、90 d硒拮抗组外周血中CD4~+淋巴细胞水平升高[(29.40±3.38)%、(45.40 ±6.01)%、(46.85 ±5.25)%,P<0.05或<0.01],60、90 d时CD4~+/CD8~+比值明显回升[(2.004±0.314)%、(2.211±0.229)%,P均<0.01].与对照组比较,30、60、90 d氟中毒组脾脏中CD4~+淋巴细胞水平降低[(47.33±5.35)%vs(41.91±4.83)%、(49.28±5.24)%vs(41.26 ±4.56)%、(34.31±4.15)%vs(29.33 ±2.89)%,P均<0.01],CD4~+/CD8~+比值也明显降低[(1.927±0.244)%vs(1.525±0.265)%、(1.847±0.224)%vs(1.640±0.198)%、(1.265±0.174)%vs(0.878 ±0.092)%,P<0.05或<0.01];与氟中毒组比较,60、90 d硒拮抗组脾脏中CD4~+淋巴细胞水平升高[(44.87±5.43)%、(32.62 ±3.37)%,P均<0.05],而30、60、90 d时CD4~+/CD8~+比值明显回升[(1.703 ±0.201)%、(1.772±0.215)%、(0.991±0.124)%,P<0.05或<0.01].30、60、90 d时氟中毒组鸡脾淋巴细胞凋亡指数[(2.31 ±0.36)%、(2.76±0.22)%、(3.04 ±0.29)%]明显高于对照组[(1.14 ±0.21)%、(1.23±0.23)%、(1.29 ±0.20)%],而60、90 d时硒拮抗组[(2.42 ±0.32)%、(2.73±0.39)%]低于氟中毒组,差异均有统计学意义(P<0.05或<0.01).结论 一定程度的硒摄入能减少氟中毒鸡淋巴细胞的凋亡、改善失衡的淋巴细胞亚群来拮抗氟的毒性.     

老年肺结核患者细胞和体液免疫状况及免疫干预治疗对其的影响

目的观察老年肺结核患者细胞及体液免疫状况,探讨免疫干预治疗对患者免疫状态的影响。方法选取80例老年肺结核患者(A组)以及80名健康老年人(B组),比较两组患者细胞及体液免疫功能指标。80例老年肺结核患者随机分为治疗组(n=40)和对照组(n=40),分别予以抗结核治疗联合免疫干预治疗和单纯抗结核治疗,比较两种方法治疗10 w后细胞及体液免疫功能指标。结果老年肺结核患者与老年健康者相比,外周血CD3+、CD4+T淋巴细胞百分比以及CD4+/CD8+比值降低〔(59.86±10.04)%vs(67.82±4.68)%,(30.64±10.93)%vs(40.87±4.78)%,1.06±0.13 vs 1.83±0.05〕,CD8+T淋巴细胞百分比较高〔(29.01±11.57)%vs(23.72±5.88)%〕(P<0.05);体液免疫功能无统计学差异(P>0.05)。80例老年肺结核患者中治疗组免疫干预治疗10 w后外周血CD3+、CD4+T淋巴细胞百分比以及CD4+/CD8+比值较对照组明显升高〔(65.73±7.21)%vs(58.93±10.20)%,(40.94±8.83)%vs(31.17±9.96)%,(1.66±0.13)vs(1.09±0.07)〕,CD8+T淋巴细胞百分比降低〔(23.01±7.19)vs(30.21±10.87)%〕(P<0.05)。结论老年肺结核患者细胞免疫功能低下,免疫干预治疗能够提高患者细胞免疫功能。     

联合谷氨酰胺和生长激素强化营养对老年危重患者细胞免疫功能的影响

目的探讨联合应用谷氨酰胺和生长激素对老年危重患者细胞免疫功能的影响。方法90例患者随机分为3组:对照组给予标准营养支持治疗,谷氨酰胺组加用谷氨酰胺,联合治疗组加用谷氨酰胺和生长激素。治疗前及治疗后7、14 d测定淋巴细胞总数、CD4/CD8及CD14单核细胞的人类白细胞抗原DR(HLA-DR)比例的变化。结果联合治疗组患者治疗14 d后淋巴细胞总数、CD4/CD8[分别为(1.92±0.38)×10~9/L、2.35±0.95]与谷氨酰胺组[(1.72±0.31)×10~9/L、2.08±0.83]及对照组[(1.28±0.24)×10~9/L、1.87±0.83]比较,差异有统计学意义(均为P<0.01);CD14单核细胞HLA-DR的表达水平[(52.5±6.8)%]与谷氨酰胺组[(42.3±6.3)%]及对照组[(34.3±4.7)%]比较亦有明显的提高(均为P<0.01);APⅡ评分及MODS评分(11.58±2.95、5.21±1.82)与谷氨酰胺组(15.33±4.25、7.26±2.12)及对照组(17.45±4.57、8.35±2.76)比较进一步下降(均为P<0.01)。结论联合谷氨酰胺和生长激素治疗能进一步提高老年危重病患者细胞免疫功能。     

多发性肌炎/皮肌炎患者外周血淋巴细胞亚群检测的临床应用价值

目的 探讨外周血淋巴细胞亚群异常在多发性肌炎和皮肌炎中的临床意义.方法 用流式细胞仪检测89例多发性肌炎／皮肌炎患者外周血淋巴细胞标记分子的表达情况,分析淋巴细胞亚群变化与疾病临床特点之间的关系.采用方差分析,独立样本t检验,x2检验及多因素Logistic回归分析进行统计学处理.结果 外周血CD3+、CD3+CD4+和CD3+CD8+细胞计数在初治并处于活动期的皮肌炎患者[分别为(8±4),(5.4±2.8)和(2.6±1.6)×108/L]明显低于非活动期皮肌炎患者[(16±6),(10.4±5.6)和(5.6±3.8)×108/L]和健康对照[(14±4),(8.3±2.8)和(4.6±1.7) ×108/L)](F=12.901,8.257,7.084;P值均＜0.05),活动期和非活动期的皮肌炎患者CD19+CD5-细胞百分比[分别为( 13.9±8.3)％和(14.2±7.0)％)]则明显高于健康对照[(6.9±2.1)％],差异具有统计学意义(F=21.443;P值均＜0.05).多因素Logistic回归分析发现,多发性肌炎/皮肌炎患者肌炎活动视觉模拟评估工具.疾病总活动度评分是患者外周血CD3+.CD3+CD4和CD3+CD8细胞计数减低的独立影响因素(b值=0.211,0.344,0.289;P值均＜0.05);多发性肌炎,皮肌炎合并间质性肺疾病(ILD)是CD3+细胞计数减低及CD3+CD4+细胞百分比减低的独立影响因素(b值=0.928,1.974;P值均＜0.05).另外Logistic回归分析发现.CD3+CD8+细胞数减低是多发性肌炎,皮肌炎死亡的独立危险因素(b值=-0.011,OR=0.989; P＜0.05).结论 外周血淋巴细胞亚群在多发性肌炎,皮肌炎疾病活动期明显异常,合并ILD可能影响部份外周淋巴细胞亚群的数量,而CD8+T细胞计数明显降低的患者死亡危险将增加.淋巴细胞亚群检测结果对临床评估多发性肌炎,皮肌炎病情活动及预后有重要指导意义.     

`粉尘螨对过敏性支气管哮喘患者气道CD~+_4CD~+_(25)T细胞的募集作用

目的 研究过敏原粉尘螨提取液能否募集CD~+_4CD~+_(25)T细胞浸润到过敏性支气管哮喘(以下简称哮喘)患者气道.方法 经纤维支气管镜将粉尘螨直接注入10例非急性发作期的轻度过敏性哮喘患者的右肺中叶或左肺舌叶的段支气管(粉尘螨侧),将生理盐水注入对侧肺段支气管作对照(对照侧).24 h后再经纤维支气管镜于相应的肺段支气管进行支气管肺泡灌洗,收集支气管肺泡灌洗液(BALF),计数BALF中的细胞数并分类.以流式细胞仪测定BALF中的T细胞及其亚群.结果 粉尘螨侧BALF中嗜酸性粒细胞[(1.4±0.1)×10~6/ml]明显多于对照侧[(0.3±0.1)×10~6/ml,P<0.003],淋巴细胞[(2.2±0.3)×10~6/ml]明显多于对照侧[(0.3±0.1)×10~6/ml,P<0.05],CD~+_4CD~+_(25)T细胞[(784.0±281.3)个/μl]明显多于对照侧[(7.7±3.6)个/μl,P<0.001].结论 粉尘螨能募集CD~+_4 CD~+_(25)T细胞浸润到缓解期轻度过敏性哮喘患者气道局部.     

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.