复元胶囊对膝骨关节炎模型血清软骨寡聚基质蛋白的影响

目的 观察复元胶囊对实验性膝骨关节炎(OA)模型病理形态及血清中软骨寡聚基质蛋白(COMP)的影响.方法 72只雄性SD大鼠随机分成正常组、模型组、壮骨关节丸组、复元胶囊组,每组18只.除正常组外,其余各组采用Hulth法建立膝OA动物模型.造模后,壮骨关节丸组、复元胶囊组分别给予壮骨关节丸、复元胶囊灌胃,而正常组、模型组给予生理盐水.灌胃后4、8、12 w分别随机选取6只进行X线摄片取材.通过X线摄片、肉眼观察、Mankin评分以及电镜检测评价复元胶囊治疗OA疗效,酶联免疫吸附法检测血清中COMP水平.结果 复元胶囊组膝关节x线积分、Mankin评分在4、8、12 w三个时间点均显著低于模型组(P<0.01),复元胶囊组血清COMP水平也低于模型组(P<0.01).结论 复元胶囊能降低血清软骨代谢标志物COMP水平,延缓OA软骨退变过程.     

复元胶囊对大鼠膝骨关节炎软骨中ADAMTS-4、5的影响

目的观察复元胶囊(FYJN)对实验性大鼠膝骨关节炎软骨组织蛋白聚糖酶1、2的影响。方法 60只雄性成年SD大鼠随机分为正常组,模型组,抗骨增生胶囊组,复元胶囊低、中、高剂量组,每组10只。除正常组外,其余各组采用Hulth法建立骨关节炎动物模型。造模1 w后,各组给予相应药物灌胃,正常组和模型组给予等体积生理盐水灌胃。治疗4 w后提取各组胫骨平台软骨组织,HE染色关节软骨Mankin评分,免疫组化法和蛋白印迹法检测关节软骨中蛋白聚糖酶1、2的蛋白表达量,并采用RT-PCR法测定组织中蛋白聚糖酶1、2的mRNA表达量。结果复元胶囊组和抗骨增生胶囊组软骨Mankin's评分明显低于模型组(P<0.01),复元胶囊高剂量组软骨组织中蛋白聚糖酶1、2的表达明显低于抗骨增生胶囊组(P<0.05)。结论复元胶囊显著降低大鼠膝关节软骨中ADAMTS-4和ADAMTS-5的表达,提示复元胶囊可能通过抑制蛋白聚糖酶,减少蛋白多糖的降解达到对早期骨关节炎防治作用。     

复元胶囊对膝骨关节炎大鼠IL-1β、OPG和OPGL mRNA表达的影响

目的观察复元胶囊对膝骨关节炎(OA)大鼠关节软骨中白细胞介素(IL)1β、骨保护素(OPG)及其配体(OPGL)的影响。方法 60只雄性SD大鼠随机分为6组:①正常组10只,不给予任何处理;②模型组10只,Hulth法造模后用生理盐水10 ml/d灌胃;③抗骨增生胶囊组10只,Hulth法造模后每日给予抗骨增生胶囊灌胃;④复元胶囊高、中、低组各10只,Hulth法造模后用不同浓度复元胶囊灌胃。于给药第12周处死大鼠,取各组大鼠膝关节内踝关节软骨标本,观察大体结构并分级;HE染色,观察其镜下结构;反转录-多聚酶链式反应(RT-PCR)法检测膝关节软骨中IL-1β、OPG及OPGL mRNA的表达含量。结果动物模型造模成功,HE染色评分中,复元胶囊高、中剂量组及抗骨增生胶囊组软骨退变程度较模型组明显减轻(P<0.01),复元胶囊组高剂量组OPG mRNA的表达量显著高于模型组(P<0.01),IL-1βmRNA及OPGL mRNA的表达显著低于模型组(P<0.05),OPG与OPGL比率显著高于模型组(P<0.01)和正常组(P<0.01)。结论复元胶囊能够上调软骨中OPG mRNA的表达,下调IL-1βmRNA及OPGL mRNA的表达,使OPG与OPGL的比率上升,表明对软骨退变及软骨下骨的破坏有一定的预防作用。     

复元胶囊对膝骨关节炎模型Ⅱ型胶原羧基端肽、基质金属蛋白酶-13的影响

目的 观察复元胶囊对实验性大鼠膝骨关节炎(KOA)模型病理形态、血清Ⅱ型胶原羧基端肽(CTX-Ⅱ)及基质金属蛋白酶-13( MMP-13)的影响.方法 72只雄性老年SD大鼠随机分成正常组、模型组、抗骨增生胶囊组、复元胶囊组(高、中、低剂量组),每组12只.除正常组外,其余各组采用Hulth法建立OA动物模型.造模2w后,各实验组给予相应的药物灌胃,正常组、模型组给予等量生理盐水.灌胃后4、8、12 w分别随机选取各组4只处死,比较各组关节软骨病理变化;免疫组化法比较MMP-13水平变化;酶联免疫吸附法(ELISA)比较CTX-Ⅱ水平变化.结果 实验组Mankin评分明显低于模型组(P＜0.01);复元胶囊中、高剂量组MMP-13及CTX-Ⅱ较模型组表达明显下调,差异有统计学意义(P＜0.05).结论 复元胶囊能够抑制关节软骨MMP-13及降解产物CTX-Ⅱ的表达,对防治关节软骨退变有一定的积极意义.     

复元胶囊对大鼠膝骨关节炎软骨iNOS及血清PGE2、NO的影响

目的 观察复元胶囊(FYJN)对实验性大鼠膝骨关节炎软骨诱导型一氧化氮合酶(iNOS)及血清一氧化氮(NO)、前列腺素E2(PGE2)的影响.方法 Hulth法建立骨关节炎模型,随机分成正常组、模型组、塞来昔布(CE)组、FYJN低、中、高剂量组,各组分别予相应药物灌胃12 w.HE染色并光镜观察股骨内侧髁软骨病理形态,免疫组化检测软骨中iNOS表达,硝酸基还原酶法及酶联免疫吸附法分别测定血清NO、PGE2含量.结果 FYJN低、中、高剂量组软骨iNOS及血清NO、PGE2均低于模型组(P<0.05,P<0.01,P<0.001).结论 FYJN呈剂量依赖关系降低骨关节炎软骨中iNOS表达及血清中NO、PGE2含量,对减轻关节炎症有一定的作用.     

复元胶囊对兔膝骨关节炎软骨中胰岛素样生长因子-I及胰岛素样生长因子结合蛋白3表达的影响

目的 观察复元胶囊对兔膝骨关节炎软骨中胰岛素样生长因子-I (IGF-I)、胰岛素样生长因子结合蛋白3(IGFBP3)表达的影响.方法 采用右后肢伸直石膏固定法造模,将44只雌雄各半新西兰大白兔随机分为正常组、模型组、盐酸氨基葡萄糖组、复元胶囊低、中、高剂量组.正常组4只,其余各组8只.各药物组每天灌胃1次,持续6 w.免疫组织化学法检测IGF-I、IGFBP3表达.结果 模型组IGF-I、IGFBP3表达积分明显高于正常组(P<0.01);复元胶囊各剂量组与模型组比较,IGF-I表达积分不同程度升高(P<0.05),而IGFBP3表达积分不同程度降低(P<0.01),高剂量组显著.结论 复元胶囊具有防治膝骨关节炎作用,其机制可能与调节IGF-I、IGFBP3表达水平有关.     

线粒体转录因子A参与软骨细胞退变逆转的机制

目的初步探讨线粒体转录因子A(TFAM)参与软骨细胞退变逆转机制,以期阐明TFAM与骨关节炎发生、发展的关系。方法以含血清培养液培养分离的原代正常软骨细胞和骨关节炎(OA)软骨细胞,MTT检测细胞存活率,流式细胞仪检测细胞凋亡变化,并利用基因组学和定量PCR等技术分析膝关节OA患者、正常关节软骨细胞的线粒体DNA特征和TFAM表达的差异情况。结果 1MTT检测OA细胞组的增殖低于正常细胞组(P<0.01);2OA组细胞凋亡率为7.11%,而正常组为1.65%,IL-1β诱导后OA细胞凋亡率为35.13%,正常组有13.79%;3OA软骨细胞TFMA mRNA的表达量低于正常软骨细胞(P<0.05),且OA软骨细胞线粒体DNA的平均拷贝数显著降低(P<0.01);线性回归分析结果表明OA细胞和正常细胞中TFMA mRNA的表达量和线粒体DNA拷贝数之间均有相关性(P<0.05)。结论分离的原代OA软骨细胞符合软骨细胞退变,为其研究提供了较好的实验对象,其线粒体DNA拷贝数与调控其转录、复制的主要调节因子TFMA的表达有相关性。     

复元胶囊对实验性骨关节炎软骨中MMP13、TIMP1及TIMP2的影响

目的观察复元胶囊对实验性骨关节炎软骨中基质金属蛋白酶13、组织型金属蛋白酶抑制剂1,2及病理形态方面的影响。方法选用新西兰兔66只,随机分为:正常对照组6只,不给予任何处理;模型对照组12只,造模后生理盐水灌胃;四环素组、复元胶囊低、中、高剂量组各12只,造模后分别用四环素、复元胶囊灌胃。连续灌胃4~12w后,比较各组关节软骨病理变化;免疫组化法检测兔膝关节软骨中MMP13、TIMP2蛋白质水平的变化;RT-PCR法检测关节软骨中MMP13、TIMP1及TIMP2mRNA表达。结果实验组软骨Mankin′s评分都显著低于模型对照组,实验组MMP13的mRNA表达水平明显低于模型对照组,而其TIMP1、TIMP2的mRNA的水平较模型对照组有所增加。MMP13主要表达于软骨表层及中上层,实验组MMP13表达量低于模型对照组(P<0.05)。结论复元胶囊能明显抑制骨关节炎软骨中MMP13的蛋白和mRNA的水平,能提高OA软骨中TIMP1,2的蛋白和mRNA的水平,稳定二者水平,对软骨退变有一定的防治作用。     

低氧诱导因子-1α对老年骨关节炎软骨细胞活性的影响

目的探讨低氧诱导因子(HIF)-1α与老年骨关节炎(OA)软骨细胞增殖和凋亡的关系。方法选择老年OA患者84例,另选健康人群80例作为对照。免疫组织化学方法检测HIF-1α、bcl-2蛋白和增殖细胞核抗原(PCNA)在OA和正常软骨细胞中的表达,分析其对OA软骨细胞增殖和凋亡的影响。结果 OA组HIF-1α阳性表达率为83.9%,明显高于正常对照组(χ2=30.312,P=0.000)。正常软骨细胞中bcl-2蛋白表达量明显高于OA软骨细胞。TUNEL法检测细胞凋亡结果显示,OA软骨组中细胞凋亡碎片明显多于正常软骨细胞,正常组软骨细胞凋亡指数为7.16±9.01,而OA组软骨细胞凋亡指数为39.7±7.23,两者差异显著(P0.05)。PCNA计数,OA组中软骨细胞增殖活性明显低于正常组。结论 HIF-1α表达与OA软骨细胞活性密切相关,可能通过调控bcl-2蛋白及PCNA影响软骨细胞的增殖与凋亡。     

基于NF-κB通路探讨鹿角壮骨胶囊对兔膝骨关节炎的保护作用机制

目的观察鹿角壮骨胶囊对兔膝骨关节炎模型核因子(NF)-κB通路的影响,探讨其对膝骨关节炎的保护作用机制。方法采用木瓜蛋白酶右侧膝关节腔内注射诱导建立兔膝骨关节炎模型,将24只长耳大白兔随机分为正常组、模型组、治疗组,正常组和模型组给予蒸馏水灌胃,治疗组给予鹿角壮骨胶囊0.3 g/(kg·d)灌胃,治疗4 w后,采用Western印迹法检测各组膝关节滑膜组织中NF-κB抑制蛋白(IκB)α、NF-κB p65、骨桥蛋白(OPN)的表达,采用免疫组化法观察各组膝关节软骨组织中血管内皮生长因子(VEGF)、低氧诱导因子(HIF)-2α、NF-κB p65的阳性表达水平。结果 Western印迹结果显示,与正常组比较,模型组兔膝关节滑膜IκBα、NF-κB p65、OPN的蛋白相对表达量明显升高(P<0.05),而治疗组IκBα、NF-κB p65、OPN的蛋白相对表达量明显低于模型组(P<0.05)。免疫组化结果显示,与正常组比较,模型组兔膝关节软骨VEGF、HIF-2α、NF-κB p65的表达明显升高(P<0.05),而治疗组VEGF、HIF-2α、NF-κB p65的表达明显低于模型组(P<0.05)。结论鹿角壮骨胶囊可能通过抑制NF-κB信号通路的激活发挥缓解膝骨关节炎的作用。     

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.