单核细胞趋化蛋白-1在BXSB狼疮小鼠肾组织中表达及药物干预效应

目的研究单核细胞趋化蛋白-1(MCP-1)在BXSB狼疮小鼠肾组织中的表达情况并探讨甲泼尼龙(MPS)是否可抑制MCP-1表达而缓解狼疮肾炎(LN)。方法应用双缩脲法、反转录聚合酶链反应(RT-PCR)、免疫组织化学法及医学图像分析系统检测各组小鼠(BXSB狼疮小鼠MPS治疗组和对照组、BALB/C正常对照组小鼠)24h尿蛋白、肾组织MCP-1的表达,并分析其与24h尿蛋白之间的相关性。结果BXSB狼疮小鼠肾组织MCP-1表达较正常BALB/C小鼠增高,以肾小管更为明显,且肾小管MCP-1的表达与24h尿蛋白呈正相关(P<0.01);甲泼尼龙干预后,MCP-1表达明显减弱(P<0.01),且肾小管MCP-1表达的下降与24h尿蛋白降低呈正相关(P<0.01)。结论MCP-1可能参与介导BXSB狼疮小鼠LN的发生发展;甲泼尼龙可通过抑制肾组织MCP-1的表达,减轻蛋白尿,可能是缓解LN作用机制之一。     

慢性移植物抗宿主病狼疮样小鼠肾组织CTGF表达及氟伐他汀的干预作用

目的观察结缔组织生长凶子(CTGF)在慢性移植物抗宿主病(cGVHD)狼疮样小鼠。肾组织中的表达情况及氟伐他汀的调节作用。方法建立cGVHD狼疮样小鼠模型。按随机设计原则将模型小鼠分为3组,即模型对照组(A组)、氟伐他汀干预组(B组)及正常对照组(C组),每组均有6只小鼠。CTGF蛋白检测采用免疫组织化学方法及免疫荧光法．mRNA表达采用反转录-聚合酶链反应(RT-PCR)检测：留取24 h尿,测定尿蛋白排泄量。结果模型对照组小鼠24 h尿蛋门总量明显高于氟伐他汀干预组(P＜0．05)及正常对照组小鼠(P＜0．01)；RT-PCR检测到模型对照组小鼠肾组织CTGF mRNA表达较氟伐他汀干预组小鼠明显升高(P＜0．05),而正常对照组小鼠表达量少；免疫组织化学及免疫荧光表明正常对照组小鼠仅在少数肾小管有微量CTGF表达,而模型对照组及氟伐他汀干预组小鼠肾小管及问质均有CTGF蛋白表达,且模型对照组小鼠表达量显著高于氟伐他汀干预组小鼠(P＜0．01)。结论CTGF可能参与介导狼疮肾炎的发生、发展；cGVHD狼疮样小鼠模型肾组织中CTGF蛋白及mRNA表达明显上调,提示CTGF可能在狼疮肾炎(LN)肾小球硬化及肾间质纤维化的进程中起重要作用；氟伐他汀可能通过降低CTGF表达及减少尿蛋白排泄从而缓解LN,延缓肾纤维化进程。     

BXSB狼疮小鼠外周血及脾肾组织干扰素-γ表达水平以及FK506对其表达的影响

目的探讨干扰素(IFN)-γ在狼疮小鼠表达和分泌的动态变化以及他克莫司(FK506)对其表达和分泌的影响.方法应用酶联免疫吸附试验(ELISA)方法和反转录-聚合酶链反应(RT-PCR)技术分别检测不同周龄BXSB小鼠(狼疮模型)和C57BL/6小鼠(正常对照)血清IFN-γ水平以及脾、肾组织IFN-γmRNA表达量,并对比了FK506处理BXSB小鼠和未处理BXSB小鼠IFN-γ表达的差异;此外本研究还检测各组动物血浆抗dsDNA抗体水平以及尿蛋白浓度.结果BXSB小鼠血IFN-γ水平以及脾、肾组织IFN-γmRNA表达量均显著高于C57BL/6小鼠;各周龄BXSB小鼠血清IFN-γ水平以及脾组织IFN-γ mRNA表达量差异无统计学意义,而16周龄以后肾组织IFN-γmRNA表达量开始显著增高.他克莫司能显著降低BXSB小鼠IFN-γ分泌水平以及睥、肾组织IFN-γmRNA表达量,也能显著降低血清抗dsDNA抗体水平以及尿蛋白浓度.结论全身循环及肾组织局部IFN-γ的高表达和高分泌在系统性红斑狼疮(SLE)的发生和发展以及肾损伤过程中起重要的作用;FK506可作为治疗SLE的免疫抑制剂之一,通过IFN-γ的作用可能是本药治疗SLE的重要作用机制之一.     

缺氧性肺动脉高压大鼠肺iNOS mRNA及蛋白表达的实验研究

目的 :通过观察慢性缺氧大鼠肺组织诱生型一氧化氮合酶 (iNOS)表达的变化 ,探讨iNOS在肺动脉高压发病中的作用。方法 :运用血清学检验及免疫组织化学斑点杂交等方法观察血一氧化氮(NO)、肺组织iNOSmRNA及蛋白表达的变化。结果 :正常组大鼠NOS表达呈弱阳性 ,慢性缺氧后肺组织iNOSmRNA升高、蛋白NOS表达呈强阳性 ,血一氧化氮水平升高 (P <0 0 1)。结论 :诱生型一氧化氮合酶、一氧化氮的增加与慢性缺氧性肺动脉高压发病有关。     

大黄和黄芪水提醇沉液注射治疗BXSB狼疮鼠的实验研究

目的 为了开发治疗系统性红斑狼疮（SLE）的新药，我们应用大黄、黄芪溶液注射治疗BXSB狼疮样小鼠，以观察其疗效。方法 应用水提醇沉法制取大黄黄芪溶液，15只BXSB雄性小鼠及6只雌鼠被随机分成三组，分别应用大黄黄芪溶液、地塞米松及生理盐水腹部皮下注射。考马斯亮蓝分光度法被用来检测小鼠尿蛋白含量，应用免疫荧光法检测小鼠血清抗核抗体（ANA）滴度及肾内免疫复合物沉积。结果 中药组及地塞米松组BXSB小鼠的尿蛋白含量及血清ANA滴度均明显低于生理盐水组（P＜0．01或P＜0．05），两组肾内免疫复合物沉积也轻于生理盐水组（P＜0．05）加药组BXSB小鼠的24h尿蛋白含量低于地塞米松组（P＜0．05）。结论 大黄、黄芪水提醇沉液可降低BXSB小鼠尿蛋白含量及血清ANA滴度，减少肾内免疫复合物沉积，对BXSB狼疮样小鼠有较为肯定的疗效。     

白杨素对MRL/lpr狼疮小鼠的保护作用及机制研究

目的：基于临床疗效确切的“多靶点”疗法(MT)及其前期疗效机制研究,探究白杨素(CHR)对狼疮性肾炎(LN)的疗效及其对白细胞介素6(IL-6)/信号传导蛋白和转录激活物3(STAT3)信号通路的影响。方法：将12只10周龄MRL/lpr狼疮小鼠随机分为疾病组及CHR治疗组,正常对照组采用C57BL/6小鼠,每组6只。CHR治疗组给予CHR 160 mg/(kg·d)灌胃,MRL/lpr组和NC组给予等量羧甲基纤维素钠(CMC-Na)灌胃,持续给药8周后处死小鼠,称量脾脏重量,评估脾脏指数;酶联免疫吸附测定(ELISA)实验检测尿蛋白/肌酐水平,过碘酸-雪夫(PAS)染色评估肾组织病理损伤程度;实时荧光定量PCR(RT-qPCR)和ELISA检测IL-6的表达和分泌水平,蛋白免疫印迹(Western Blot)和免疫组化染色(IHC)评估肾组织中STAT3蛋白的磷酸化水平。结果：CHR治疗可下调MRL/lpr狼疮小鼠脾重及脾脏指数、抑制尿蛋白/肌酐水平并缓解肾脏病理损伤。此外CHR能够显著抑制IL-6分泌、表达水平及肾组织p-STAT3蛋白表达。结论：CHR对MRL/lpr狼疮小鼠具有...     

三氧化二砷对BXSB狼疮小鼠神经激肽A的影响及其意义探讨

目的 探讨不同浓度三氧化二砷(As2O3,ATO)对BXSB狼疮小鼠肾组织中神经激肽A(NKA)含量的影响及其意义.方法 BXSB狼疮小鼠50只随机分为5组,分别为对照组,系统性红斑狼疮(SLE)组,低剂量、中剂量和高剂量ATO组.常规检验治疗前后小鼠的血生化指标、组织化学法观察肾脏病理改变、酶联免疫吸附试验(ELISA)及反转录一聚合酶链反应(RT-PCR)法检测各组NKA及肾组织中NKA mRNA含量.结果 BXSB狼疮小鼠肾组织中NKA的含量(299±26)pg/g高于健康对照组(122±7)Dg/g,差异有统计学意义(P＜0.05).ATO作用BXSB狼疮小鼠后,其肾组织的NKA含量明显下降,大、中、小剂量组与治疗前的差异均具有统计学意义(P＜0.05).同时,各项生化指标及肾脏病理改变也明显改善(P＜0.05),尤其以小剂量ATO组效果更好(P＜0.01);小剂量ATO组中毒症状较轻,中、大剂量组中毒症状较重,两者差异有统计学意义(P＜0.05).结论 一定剂量的ATO对狼疮肾具有治疗作用,其机制可能是下调肾组织中NKA mRNA表达,其中小剂量ATO治疗狼疮肾,既安全又有效,具有一定的应用前景.     

精氨酸升压素对大鼠心肌成纤维细胞诱导型一氧化氮合酶-一氧化氮系统活性的影响

目的　研究精氨酸升压素 (AVP)对大鼠心肌成纤维细胞诱导型一氧化氮合酶 (iNOS) 一氧化氮 (NO)系统活性的影响。方法　胰酶消化法分离培养Sprague Dawley仔鼠心肌成纤维细胞 ,采用硝酸还原酶法、蛋白质印迹和逆转录 聚合酶链式反应观察AVP对心肌成纤维细胞的NO含量、iNOS蛋白水平和iNOSmRNA表达的影响。结果　 (1)不同浓度AVP干预下 ,心肌成纤维细胞的NO含量、iNOS蛋白水平和iNOSmRNA表达都随AVP浓度的增高而增加。其中 10 -7mol/LAVP组和 10 -6mol/LAVP组的NO含量 [(6 9 0 5± 5 5 6 ) μmol/L和 (6 2 86± 6 0 5 ) μmol/L]、iNOS蛋白水平 (0 73± 0 0 6和 0 6 4± 0 0 5 )和iNOSmRNA表达 (0 70± 0 0 3和 0 6 6± 0 0 6 )都显著高于对照组 [(2 9 34± 5 34)μmol/L ,0 2 0± 0 0 5 ,0 2 5± 0 0 4 ]、10 -9mol/LAVP组 [(31 79± 6 5 9) μmol/L ,0 2 4± 0 0 7,0 30±0 0 6 ]和 10 -8mol/LAVP组 [(36 87± 7 89) μmol/L ,0 30± 0 0 9,0 31± 0 0 3]。但 10 -6mol/LAVP组的NO含量、iNOS蛋白水平和iNOSmRNA表达都低于 10 -7mol/LAVP组。 (2 ) 10 -7mol/LAVP干预下CFs的NO含量、iNOS蛋白水平和iNOSmRNA表达都随培养时间的延长而增加。其中 2 4h组和 36h组的NO含量 [(6 5     

p38 MAPK在内毒素休克小鼠肺组织诱导型一氧化氮合酶表达中的作用

目的　观察脂多糖 (LPS)诱导的小鼠肺组织诱导型一氧化氮合酶 (iNOS)表达的变化趋势 ,探讨p38丝裂原激活的蛋白激酶 (p38MAPK)在内毒素休克发生中的作用。方法　复制小鼠内毒素休克动物模型 ,分别观察LPS作用和组织中iNOS表达的时间过程、剂量 效应关系及 p38MAPK在其中的作用。动物在LPS处理后 ,收集血液 ,用Griess法检测血浆一氧化氮 (NO)水平 ;取肺、肝、肾等组织样本 ,分别用Western印迹法和RT PCR检测组织iNOS蛋白和mRNA表达 ,用免疫沉淀和放射自显影检测组织p38MAPK活性。结果　①腹腔注射LPS后可引起双峰型进行性的动脉血压下降 ,6h后达到重度休克 ;②iNOS在正常肺中就有少量表达 ,注射内毒素后肺组织iNOSmRNA和蛋白表达增加 ,血浆中NO水平升高 ,它们与LPS存在剂量 效应关系 ,并且在一定的时间范围内 ,随着LPS作用时间的增加iNOS表达也增高 ;③注射LPS后 ,在心、肝、脾、肺、肾、肠等组织中 ,肺iNOS表达最早 ,水平最高 ;④LPS注射的小鼠iNOS表达增高的同时 ,肺中 p38MAPK信号转导通路被激活 ;用p38MAPK特异性抑制剂SB 2 0 3580后 ,可以显著降低血浆NO水平及肺组织中iNOSmRNA和蛋白的表达 ,但对血压下降并无显著性影响。结论　在正常情况下 ,小鼠肺组织有少量iNOS表达 ,内毒素休克时全身多种器     

三氧化二砷对MRL/lpr狼疮小鼠肾脏病理和组织转化生长因子-β1核因子-κBp65异常表达的影响

目的 研究MRL/lpr狼疮小鼠肾脏病理转化生长因了(TGF)-β1、核因子(NF)-κBp65的异常表达及三氧化二砷(ATO)的干预作用.方法 将MRL/lpr狼疮鼠45只随机分为ATO组、环磷酰胺(CTX)组和对照组.观察各组MRL/lpr狼疮小鼠治疗前后尿蛋内(半定量)的变化;给药2个月后处死,经苏木素-伊红(HE)染色、PAS染色后观察肾组织病变程度;用免疫荧光方法检测肾组织IgG、C3的表达;用免疫组织化学方法检测肾组织TGF-β1、NF-κBp65的表达.结果 ①尿蛋白分析:给药后ATO组较对照组有明显减少(P=0.004)、CTX组较对照组也有减少(P=0.005).②免疫荧光检测IgG、C3的表达:在对照组可见IgG沿肾小球系膜区,毛细血管袢弥漫沉积(+～+++),而ATO组、CTX组则明显减少(-～+),经统计发现ATO组、CTX组较对照组明显减少(P＜0.05);C3的表达差异无统计学意义.③肾小球细胞数及肾组织活动积分在ATO组及CTX组中均较对照组明显减少(P＜0.05).④免疫组织化学:TGF-β1、NF-κBp65在对照组肾小管上皮细胞可见明显表达,经ATO及CTX组表达明显减少,肾小球表达各组均不明显.结论 TGF-B1、NF-κBp65主要表达在MRL/lpr狼疮小鼠肾小管上皮细胞,ATO通过下调TGF-β1、NF-Kκp65的表达改善小管病变达到改善狼疮肾炎的进展.     

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.