狼疮肾炎患者肾组织中结缔组织生长因子的转录表达及意义

目的探讨人类狼疮肾炎(LN)患者肾组织中结缔组织生长因子(CTGF)的基因转录和蛋白表达与肾纤维化之间的关系。方法应用免疫组织化学和原位杂交技术,以15例正常肾组织为对照,观察了36例LN患者肾组织中CTGF表达的分布和表达量的变化,分析肾组织中CTGF表达与LN肾间质病变的内在联系。结果LN患者肾组织的CTGF表达主要分布在肾小管上皮细胞和肾间质细胞、肾小球脏层和壁层上皮细胞和系膜区细胞。正常肾组织没有或只有极弱的CTGFmRNA转录和蛋白表达;LN患者肾组织CTGFmRNA转录及蛋白表达量较正常肾组织显著增高(CTGFmRNA:8.22±3.13比1.12±0.23;CTGF蛋白:8.74±3.24比1.21±0.22,P均<0.001);LN患者肾组织CTGF表达量与肾间质病变程度呈正相关(r=0.863,P<0.01);Ⅳ型LN患者肾组织CTGF表达量显著高于非Ⅳ型LN患者(CTGFmRNA:10.73±3.61比4.78±1.82CTGF蛋白:11.03±1.12比5.02±1.31,P均<0.01);CTGF的蛋白表达量与转化生长因子(TGF)-β1、纤维结合蛋白(Fn)及ColⅢ的表达量呈正相关(r=0.704、r=0.783及r=0.756,P均<0.01)。结论CTGFmRNA转录和蛋白表达量的增加与LN患者肾小管间质病变程度加重呈正相关。作为TGF-β的下游介质,CTGF可通过促进细胞外基质如FN和ColⅢ等合成,从而在LN患者肾小球硬化及肾小管间质纤维化的     

狼疮肾炎患者肾组织中吲哚胺2,3-双加氧酶的表达及意义

目的检测Ⅳ型狼疮肾炎（LN）患者肾组织中吲哚胺2，3-双加氧酶（IDO）的表达情况及其与肾间质浸润炎细胞的增殖和肾小管-间质病理损害程度之间的关系。方法采用免疫组织化学染色法观察正常肾组织和Ⅳ型LN患者肾组织IDO的表达情况，并进一步分析其表达与肾间质浸润核增殖抗原（PCNA）阳性淋巴细胞数及肾小管-间质（TIL）病理损害程度之间的相关关系。结果正常肾组织未检测到IDO表达，而在Ⅳ型LN肾小管上皮细胞IDO的表达阳性，并且LN肾小管上皮细胞表达IDO的阳性强度与TIL PCNA＋浸润细胞数及TIL病理损害程度呈显著负相关。结论IDO在Ⅳ型LN肾小管上皮细胞中呈高表达，并与TIL浸润细胞增殖情况及TIL病理变化呈负相关，提示IDO可能参与LNTIL病理损害过程。     

KIM-1在TGF-β1诱导的大鼠肾小管上皮细胞纤维化因子表达中的作用

目的探讨肾脏损伤因子(KIM)-1在转化生长因子(TGF)-β1诱导的肾小管上皮细胞纤维化因子表达中的作用。方法用TGF-β1刺激处理大鼠肾小管上皮细胞NRK52E,qRT-PCR和Western印迹法分别测定细胞中KIM-1 mRNA和蛋白水平。在NRK52E中转染KIM-1 siRNA,给予TGF-β1刺激后,qRT-PCR和Western印迹法分别测定细胞中KIM-1 mR-NA和蛋白水平。Western印迹法检测细胞中上皮间质转化(EMT)标志蛋白上皮钙黏附素(E-cadherin)、α-平滑肌肌动蛋白(SMA)和纤维化因子结缔组织生长因子(CTGF)、1型胶原酶(Col1)、3型胶原酶(Col3)的表达。结果对照组+TGF-β1细胞中KIM-1 mRNA和蛋白水平明显高于对照组(P<0. 05)。干扰组+TGF-β1细胞中KIM-1表达水平显著低于对照组+TGF-β1,差异具有统计学意义(P<0. 05)。干扰组+TGF-β1细胞中E-cadherin蛋白表达水平升高,α-SMA表达水平降低,纤维化因子CTGF、Col1、Col3的表达也降低,与对照组+TGF-β1比较差异具有统计学意义(P<0. 05)。结论敲低KIM-1可以减少TGF-β1诱导的肾小管上皮细胞纤维化因子表达,抑制肾小管上皮细胞EMT。     

过氧化物酶体增殖物激活受体γ激动剂对TGF-β1诱导肾成纤维细胞细胞外基质表达的影响

目的:研究过氧化物酶体增殖物激活受体γ(PPARγ)激动剂对转化生长因子β1(TGF-β1)致肾间质纤维化作用的影响,探讨其抗肾间质纤维化的潜在作用.方法:体外培养大鼠肾成纤维细胞株(NRK/49F),利用逆转录-聚合酶链反应(RT-PCR)观察PPARγ配体15d-PGJ2及其激动剂曲格列酮和齐格列酮对TGF-β1诱导的α-平滑肌肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)、纤维连结蛋白(FN)和Ⅲ型胶原(ColⅢ)mRNA表达的影响,利用Western Blot方法观察PPARγ激动剂对TGF-β1诱导的FN蛋白表达的影响.结果:TGF-β1能显著增加NRK/49F细胞CTGF、FN和ColⅢmRNA表达,并呈剂量依赖和时间依赖效应.与TGF-β1刺激组相比,10 μM 15d-PGJ2、曲格列酮和齐格列酮预处理组α-SMA、CTGF、FN和ColⅢmRNA表达量显著减少,FN蛋白表达量显著下降.结论:PPARγ激动剂可抑制TGF-β1诱导的肾间质成纤维细胞CTGF表达和细胞外基质(ECM)合成.     

狼疮性肾炎患者肾组织白介素-10的表达与巨噬细胞浸润的关系及其意义

目的:研究白介素-10(IL-10)在狼疮性肾炎(LN)患者肾组织中表达及巨噬细胞的浸润与肾脏病理和功能损害的关系.方法:采用微波免疫组织化学双重染色法测定正常肾组织和20例LN肾组织巨噬细胞浸润和IL-10的表达及与狼疮肾组织活动指数、肾脏组织学指标和功能损害的相关关系.结果:①正常肾小球及肾小管间质仅个别巨噬细胞存在,几乎无IL-10表达,而在各型LN,肾小球和肾小管间质见浸润的巨噬细胞及IL-10阳性细胞明显增多,且病变严重的Ⅳ型LN尤为明显.②肾小球KP1 巨噬细胞与肾小球细胞数增多,LN活动指数、慢性指数均呈显著相关,IL-10 细胞数也与这些指标相关.肾小管间质中KP1 巨噬细胞数及IL-10 细胞与小管损害、狼疮肾组织活动指数和慢性指数(特别是慢性指数)显著正相关.③肾小球肾小管间质中KP1 细胞数与24h尿蛋白排泄量、尿NAG排泄量有关,IL-10 细胞也与上述指标有关,而与血清学免疫指标无关.④肾小球KP1 与IL-10 细胞呈高度相关性,r=0.79(P＜0.001),小管间质中KP1 与IL-10 细胞也呈高度相关性:r=0.76(P＜0.001).70%KP1 细胞聚集在肾组织损害严重部位,同时IL-10阳性染色物主要存在于KP1 细胞中.结论:肾脏IL-10表达增加与巨噬细胞浸润在LN病理与功能损害中起重要作用.     

PPARγ在狼疮肾炎患者肾组织中的表达及其意义

目的观察过氧化物酶体增殖物激活受体γ(PPARγ)在狼疮肾炎(LN)不同病变类型肾组织中的表达及其与肾脏病理改变之间的关系,探讨PPARγ在LN发病机制中的可能作用。方法根据临床及肾活检病理诊断,选择LN不同病变类型患者作为研究对象,其中Ⅱ型(6例),Ⅳ型(8例),Ⅴ型(7例)。以肾脏肿瘤切除术中远离肿瘤部位的正常肾组织(6例)为对照。光镜下观察并计数肾脏病变活动指数、间质病变指数；用免疫组织化学方法对各例肾组织PPARγ的表达进行检测,并对其与肾脏病变的相关性进行分析。结果LN患者肾组织。肾小管、肾小球及间质浸润细胞中PPARγ脚表达较正常对照组均显著上调,其中Ⅳ型LN肾组织肾小管、肾小球及间质浸润细胞PPARγ表达显著高于Ⅱ型、Ⅴ型；肾小球PPARγ染色阳性细胞数目与肾脏病理活动积分及间质病变指数之间呈显著正相关(r=0．94；P＜0．01)。结论PPARγ在LN肾组织肾小管、肾小球及间质浸润细胞高表达对于限制LN肾脏病变发展可能具有重要作用     

原发性IgA肾病中肥大细胞浸润与肾小管间质损害的关系

目的:了解肥大细胞(mast cell,MC)在人类原发性IgA肾病(IgAN)肾组织中的浸润及与肾小管间质损害程度、蛋白酶激活受体2(protease activated receptor-2 PAR-2)表达的关系,探寻肥大细胞在IeAN肾小管间质损害过程中的可能作用机制. 方法:以35例IgAN肾活检石蜡包埋肾组织及5例肾肿瘤患者行肾切除的远离肿瘤部位的肾组织作为研究对象.根据Lee氏分级及Katafuehi半定量积分法分组.采用免疫组化法检测肾组织中MC和PAR-2的表达,分析它们之间及其与肾小管问质损害程度的相关性.所有数据均使用SPSS 13.0英文版统计软件进行分析. 结果:(1)随着Lee氏分级等级的升高和肾小管间质损害程度的加重,MC数目和PAR-2的表达水平逐渐增加.(2)肾小管间质中MC、PAR-2的表达水平呈正相关(r=0.844,P<0.01).肾小管间质中MC和PAR-2的表达水平均与肾小管间质病理积分呈正相关(r=0.948,0.840,P均<0.01). 结论:(1)MC浸润与原发性IgAN肾小管间质损害程度密切相关,MC可能是原发性IgAN肾小管间质损害过程的重要参与者.(2)类胰蛋白酶和PAR-2可能共同参与了原发性IgAN复杂的肾小管间质损害过程.     

肾组织核心蛋白聚糖和转化生长因子-β1的表达与肾间质纤维化的关系

目的探讨转化生长因子β 1(TGF-β1)及核心蛋白聚糖(DCN)表达与肾间质纤维化之间的关系,及α- 平滑肌肌动蛋白(α-SMA)和单核巨噬细胞浸润在肾脏疾病中的变化. 方法收集30例不同肾脏病肾活检患者的临床和病理资料,应用免疫组织化学染色方法,检测TGF-β1及DCN在肾间质内的表达,以及α-SMA和CD 68的表达.并分析它们的表达与肾间质纤维化之间的关系. 结果肾间质病变严重程度与血清肌酐、肾内TGF-β1和DCN表达程度成正相关,与内生肌酐清除率成负相关.肾间质浸润的巨噬细胞主要见于间质大量炎性细胞浸润的部位及纤维化间质和病变肾小球周围;α-SMA表达可见于血管壁、纤维化间质、增厚的肾小球囊壁或病变肾小球周围以及变性萎缩的肾小管管周,它们的表达与肾间质纤维化程度呈正相关. 结论随着肾间质纤维化程度的加重,DCN和 TGF-β1的表达逐渐增加,同时伴有间质单核巨噬细胞的浸润和间质细胞的激活,参与肾间质纤维化的进程.     

Ⅳ型狼疮肾炎患者肾组织中程序性死亡配体-1-程序性死亡-1的表达及其意义

目的 了解Ⅳ型狼疮肾炎(LN)患者肾组织中程序性死亡配体-1(PD-L1)、程序性死亡-1(PD-1)的表达及PD-L1与PD-1、肾小管间质病理损害程度的关系、方法 采用免疫组织化学、原位杂交染色法，观察正常肾和Ⅳ型LN患者肾组织PD-L1、PD-1的表达，以及PD-L1阳性强度与肾小管—间质PD-1阳性细胞数、小管—间质病理损害程度的关系。结果 Ⅳ型LN肾组织表达PD-L1、PD-1较正常组增强：狼疮组肾小管PD-L1阳性强度与肾问质PD-1阳性细胞数呈正相关，与肾小管间质病理损害程度呈负相关。结论PD-L1～PD-1信号通路在Ⅳ型狼疮性肾炎肾小管间质病理损害中起重要作用。     

过氧化物酶体增殖物激活受体γ激动剂对TGF-β1诱导肾成纤维细胞细胞外基质表达的影响水

目的：研究过氧化物酶体增殖物激活受体γ（PPARγ）激动剂对转化生长因子β1（TGF-β1）致肾间质纤维化作用的影响，探讨其抗肾间质纤维化的潜在作用。方法：体外培养大鼠肾成纤维细胞株（NRK／49F），利用逆转录-聚合酶链反应（RT-PCR）观察PPARγ配体15d-PGJ2及其激动剂曲格列酮和齐格列酮对TGF-β1诱导的α-平滑肌肌动蛋白（α-SMA）、结缔组织生长因子（CTGF）、纤维连结蛋白（FN）和Ⅲ型胶原（ColⅢ）mRNA表达的影响，利用Western Blot方法观察PPARγ激动剂对TGF-β1诱导的FN蛋白表达的影响。结果：TGF-β1能显著增加NRK／49F细胞CTGF、FN和ColⅢmRNA表达，并呈剂量依赖和时间依赖效应。与TGF-β1刺激组相比，10μM 15d-PGJ2、曲格列酮和齐格列酮预处理组叶SMA、CTGF、FN和ColⅢmRNA表达量显著减少，FN蛋白表达量显著下降。结论：PPA脚激动剂可抑制TGF-β1，诱导的肾间质成纤维细胞CTGF表达和细胞外基质（ECM）合成。     

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.