罗红霉素和地塞米松对支气管哮喘大鼠气道重塑中成肌纤维细胞的影响

目的探讨成肌纤维细胞（myofibroblast,MF）在支气管哮喘（简称哮喘）气道重塑中的作用。观察罗红霉素对哮喘气道重塑的影响,并与地塞米松作对照。方法SD大鼠40只,随机分为哮喘组（A组）、生理盐水对照组（C组）、地塞米松治疗组（D组）和罗红霉素治疗组（R组）,每组10只。利用卵白蛋白（ovalbumin,OVA）/Al（OH）3致敏与OVA雾化吸入激发建立大鼠哮喘模型。免疫组织化学测定肺组织中支气管上皮下MF的α-平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）表达含量,并使用图象分析技术进行积分光密度（integral optical density,IOD）定量分析测定。光镜观察肺组织病理结构变化,图像分析软件分析,并测定肺内支气管总管壁厚度、内管壁厚度、平滑肌层厚度等指标。结果免疫组织化学和图像分析结果：A组与C组相比,总管壁厚度、内管壁厚度、平滑肌层厚度显著增厚（P〈0.01）。D组和R组中的内管壁厚度、平滑肌层厚度与A组相比,均显著变薄（P〈0.01）。R组的总管壁厚度、内管壁厚度、平滑肌层厚度与D组相比差异无统计学意义。定量分析测定的IOD值显示A组支气管上皮下MFα-SMA表达含量较C组显著增加（P〈0.01）,D组和R组表达含量较A组均减少（P〈0.05）,R组表达含量与D组相比差异无统计学意义。相关分析结果：支气管上皮下MFα-SMA表达含量（用IOD值表示）和内管壁厚度呈正相关（r=0.913,P〈0.01,n=40）,和平滑肌层厚度也呈正相关（r=0.626,P〈0.01,n=40）。结论MF在气道重塑形成中起重要作用。罗红霉素和地塞米松均可能通过抑制MF增殖和表达起到抗哮喘气道重塑作用。     

L型钙离子通道阻滞剂非洛地平对大鼠哮喘气道重塑模型STAT6、SM-α-actin影响研究

目的通过卵蛋白致敏大鼠建立支气管哮喘（简称哮喘）气道重塑模型,探讨非洛地平对气道壁信号转导和转录激活因子6（STAT6）、α平滑肌肌动蛋白（SM-α-actin）在哮喘气道重塑模型中表达的影响与哮喘气道平滑肌细胞（airway smooth muscle cell,ASMC）内钙稳态失衡与哮喘气道炎症的病理机制。方法卵蛋白诱发大鼠慢性哮喘模型：30只SD大鼠,按随机数字法分为三组,对照组（A）、哮喘组（B）、非洛地平组（C）,B组和C组用10％卵蛋白氢氧化铝混合液腹腔注射致敏,1％卵蛋白雾化吸人激发,c组激发前给予非洛地平灌胃,进行4周干预。大鼠肺组织进行病理组织学图像分析,STAT6、SM-α-actin原位表达研究分析采用免疫组织化学技术。结果免疫组织化学染色：STAT6在A组气道壁有少量表达,B组主要表达在黏膜下层,IOD值测定B组和A组比较表达量显著增多（P〈0．01）,C组和B组比较表达量明显减少（P〈0．01）。结论①卵蛋白4周激发致敏SD大鼠可建立慢性哮喘气道重塑模型。②STAT6、SM-α-actin在慢性哮喘气道黏膜下层表达明显增多。③L型钙离子通道阻滞剂非洛地平可减少哮喘气道壁STAT6、SM-α-actin的表达。     

川芎嗪对哮喘大鼠转录因子GATA-3表达的影响

目的 观察川芎嗪对哮喘大鼠转录因子GATA-3表达的影响。方法 72只SPF级SD大鼠随机分为正常对照组（A组）、哮喘模型组（B组）、川芎嗪小剂量组（C组,20mg／kg）、川芎嗪中剂量组（D组,40mg／kg）、川芎嗪大剂量组（E组,80mg／kg）和地塞米松组（F组）,每组12只。以卵蛋白腹腔注射并雾化吸入制备大鼠哮喘模型,用动物呼吸机测定气道反应性评价造模效果。采用免疫组化半定量法测定肺组织GATA-3含量。结果 给予乙酰胆碱（Ach）激发后,比较各组大鼠呼气相气道阻力（Re）,显示造模成功;B、C、D、E和F组的GATA-3表达量显著高于A组,其差异均有统计学意义（P〈0．01）;C、D、E和F组的GATA-3表达量和B组比较,差异均有统计学意义（P〈0．01）;川芎嗪剂量的增加与GATA-3的表达呈负相关趋势;两两比较E组和F组差异无统计学意义。结论哮喘大鼠存在GATA-3高表达;川芎嗪减低气道高反应性,抑制GATA-3的表达,纠正Th1／Th2失衡,从而治疗哮喘。     

糖皮质激素对哮喘气道重塑大鼠骨桥蛋白水平及骨桥蛋白mRNA表达的影响

目的探讨糖皮质激素对哮喘气道重塑大鼠骨桥蛋白(OPN)水平及OPN mRNA表达的影响。方法2014年12月—2016年1月,选取30只雄性Wistar大鼠并采用随机数字表法分为正常对照组、哮喘模型组、糖皮质激素组,各10只。建立哮喘动物模型,哮喘模型组大鼠于造模第15天予以卵清蛋白(OVA)雾化吸入激发致敏;正常对照组大鼠于造模第15天予以等量0.9%氯化钠溶液腹腔注射及雾化吸入;糖皮质激素组大鼠处理同哮喘模型组,并于OVA雾化吸入激发致敏前1 h雾化吸入布地奈德混悬液(BUD)。比较3组大鼠气道壁厚度,支气管壁平滑肌厚度及胰岛素样生长因子-I(IGF-I)、神经生长因子(NGF)、核因子-κB(NF-κB)水平,OPN水平及OPN mRNA表达情况,并进行相关性分析。结果哮喘模型组大鼠气道壁厚度、支气管壁平滑肌厚度大于糖皮质激素组和正常组对照组(P0.05);糖皮质激素组大鼠气道壁厚度、支气管壁平滑肌厚度大于正常组对照组(P0.05)。哮喘模型组大鼠IGF-I、NGF、NF-κB水平高于糖皮质激素组和正常组对照组(P0.05);糖皮质激素组大鼠IGF-I、NGF、NF-κB水平高于正常组对照组(P0.05)。哮喘模型组大鼠OPN水平和OPN mRNA相对表达量高于糖皮质激素组和正常组对照组(P0.05);糖皮质激素组大鼠OPN水平和OPN mRNA相对表达量高于正常组对照组(P0.05)。哮喘气道重塑大鼠气道壁厚度、支气管壁平滑肌厚度及IGF-I、NGF、NF-κB水平分别与OPN水平(r值分别为0.91、0.84、0.80、0.79、0.83)、OPN mRNA相对表达量(r值分别为0.83、0.87、0.79、0.76、0.81)呈正相关(P0.05)。结论糖皮质激素可抑制哮喘大鼠气道重塑的发生,延缓气道壁及支气管壁平滑肌增厚,降低IGF-I、NGF、NF-κB、OPN水平及OPN mRNA的表达。     

咳喘宁对支气管哮喘大鼠气道重塑及MMP-9、TIMP-1的影响

目的 观察咳喘宁对支气管哮喘大鼠气道形态学和基质金属蛋白酶-9（MMP-9）、基质金属蛋白酶组织抑制剂-1（TIMP-1）的影响.方法 40只SD大鼠随机分为正常组、模型组、咳喘宁高、低剂量组（27 和13.5 g生药/kg体重）、桂龙咳喘宁胶囊对照组（0.41 g/kg体重）,每组8只.除正常组外以卵蛋白致敏并吸入激发法制备大鼠支气管哮喘模型,各治疗组均从第1次哮喘激发开始（造模第3周）至处死前每天灌胃给药,激发并给药4 w后处死大鼠,取肺组织HE染色,彩色图像分析仪测量支气管管壁厚度、平滑肌厚度,采用ELISA双抗体夹心法测定肺组织MMP-9、TIMP-1含量.结果 与正常组比较,模型组大鼠支气管管壁和平滑肌厚度、肺组织MMP-9、TIMP-1含量及二者比值明显增加（P＜0.01）;与模型组比较,各治疗组均可显著降低支气管管壁和平滑肌厚度（P＜0.01）,降低肺组织MMP-9、TIMP-1含量以及二者比值（P＜0.05或P＜0.01）;且咳喘宁高、低剂量组优于桂龙咳喘宁胶囊组（P＜0.05或P＜0.01）.结论咳喘宁可通过降低肺组织MMP-9和TIMP-1含量,调节二者比值,抑制支气管哮喘大鼠气道壁增厚和平滑肌增生肥大,从而抑制支气管哮喘大鼠气道重塑.     

雷公藤甲素对哮喘大鼠气道平滑肌增生及c-fos与c-jun表达的影响

目的：探讨雷公藤甲素（TL）对哮喘大鼠气道平滑肌（ASM）增生及原癌基因c-fos与c-jun基因表达的影响。方法：选择70只雄性SD大鼠随机分为7组，每组10只，采用卵蛋白腹腔注射及雾化吸入制成大鼠哮喘模型，采用志录－聚合酶链反应（RT－PCR）法检测各组的c-fos与c-jun的mRNA表达水平，同时用免疫组化方法检测c-fos与c-jun蛋白表达水平，并在光镜下观察支气管壁厚度（WA／Pi)，支气管平滑肌厚度（平滑肌面积／Pi)，支气管平滑肌细胞核数量（N／Pi)及肺组织形态学改变，结果：RT－PCR和免疫组化均显著哮喘组（A组），治疗组（C，D，E，F，G组）c-fos与c-jun的mRNA和蛋白表达与对照组（B组）比较，差异有显著性（P＜0．01）；各治疗组的表达与A组比较差异也有显著性（P＜0．01），而各治疗组之间比较差异无显著性（P＞0．05），是结果显示，TL减轻ASM的增生。结论：ASM增生肥厚是哮喘的一个显著特征，c-fos及c-jun可能在此起着促进作用，TL通过下调c-fos和c-jun表达而起抑制ASM增生作用。     

5-羟色胺在支气管哮喘豚鼠气道重塑中的作用机制研究

目的 探讨5-羟色胺（5-HT）在支气管哮喘（下称哮喘）豚鼠气道重塑中的作用机制。方法 将50只雄性豚鼠随机分为正常对照组（A组）、哮喘模型组（B组）、哮喘模型延续组（B1组）、5-HT组（B2组）、5-HT拮抗剂组（B3组）。用卵清蛋白复制哮喘气道重塑模型。观察血清5-HT浓度和气道壁各层厚度。结果 ①血清5-HT浓度与气道壁各层厚度B组明显大于A组,B1组明显大于B组,P〈0．05,〈0．01。②气道壁各层厚度B2组大于B1组,B3组小于B1组,P均〈0．05。结论 5-HT浓度变化是哮喘豚鼠气道重塑的机制之一。     

豚鼠中性粒细胞性哮喘模型的建立

目的建立豚鼠中性粒细胞性哮喘模型。方法 40只成年雄性豚鼠按随机数字表法分为对照组（A组）、嗜酸粒细胞性哮喘组（B组）、中性粒细胞性哮喘组（C组）、嗜酸粒细胞性哮喘治疗组（D组）和中性粒细胞性哮喘治疗组（E组）。用卵白蛋白（ovalbumin,OVA）和弗氏完全佐剂（Freund’s complete adjuvant,FCA）联合致敏或OVA单独致敏豚鼠后用OVA雾化吸入激发建立中性粒细胞性哮喘或嗜酸粒细胞性哮喘模型,治疗组在激发前腹腔注射地塞米松。观察各组豚鼠雾化激发后体征变化及支气管肺组织病理改变,并比较各组豚鼠气道阻力、血中白细胞分类计数、支气管肺泡灌洗液（brochial alveolar lavage fiuid,BALF）细胞总数及分类计数。结果 B、C组豚鼠激发后均出现典型哮喘症状,不同浓度乙酰甲胆碱激发后的气道阻力与A组相比均显著增高（P〈0.05）;B组豚鼠BALF总数、BALF及血中嗜酸粒细胞所占比例与A组比较显著增加（P〈0.05）;C组豚鼠BALF总数、BALF及血中中性粒细胞所占比例与A组比较均显著增加（P〈0.05）;C组豚鼠BALF中性粒细胞所占比例与B组相比显著增加（P〈0.05）;除B、D组血中性粒细胞外,D和E两组的上述其他各项指标分别与B组和C组相比明显降低（P〈0.05）,以上差异均有统计学意义。B、C两组豚鼠支气管肺组织病理均提示支气管管腔狭窄、黏膜上皮脱落、炎症细胞浸润等典型的哮喘病理学改变,其中B组以嗜酸粒细胞浸润为主,C组以中性粒细胞浸润为主,D、E组较之明显好转。结论本实验建立的豚鼠中性粒细胞性哮喘模型是成功的。     

L-精氨酸对哮喘大鼠气道平滑肌细胞增殖的影响

目的 观察L-精氨酸（L-Arg）对哮喘气道重构大鼠气道平滑肌细胞（ASMC）的细胞周期分布及细胞周期调节蛋白（CCRP）表达水平的影响,探讨L-Arg体内干预哮喘大鼠ASMC增殖的可能作用机制。方法实验分成对照组、哮喘组、L-Arg组,建立大鼠哮喘气道重构模型,检测血清NO2^-／NO3^-含量、肺内支气管内管壁和平滑肌层厚度及ASMC核数、ASMC的细胞周期分布以及ASMC内细胞周期素E（cyclin E）、cyclinA、cyclinB、蛋白27^kip1（P27^kip1）的表达。结果 哮喘组大鼠支气管内管壁、平滑肌层的厚度和ASMC数目显著大于对照组（P〈0．05）;L-Arg组大鼠支气管内管壁的厚度、平滑肌层的厚度和ASMC数目显著小于哮喘组（P〈0．05）。哮喘组血清一氧化氮（nitricoxide,NO）水平显著低于对照组（P〈0．05）;L-Arg组血清NO水平显著高于哮喘组（P〈0．01）。哮喘组G0／G1期ASMC比例及P27^kip1表达水平显著低于对照组（P〈0．01）,G2／M＋S期ASMC比例及cyclinE、cyclinA和cyclinB表达水平显著高于对照组（P〈0．01）;L-Arg组G0／G1期ASMC比例及P27^kip1表达水平显著高于哮喘组（P〈0．05）,G2／M＋S期ASMC比例及cyclinE和cyclinA表达水平显著低于哮喘组（P〈0．05）。结论 L-Arg通过调控CCRP的表达水平阻滞细胞从G1期进入S期而抑制哮喘ASMC的增殖。     

罗红霉素和地塞米松对支气管哮喘大鼠气道重塑中成肌纤维细胞的影响

目的 探讨成肌纤维细胞(myofibroblast,MF)在支气管哮喘(简称哮喘)气道重塑中的作用.观察罗红霉素对哮喘气道重塑的影响,并与地塞米松作对照.方法 SD大鼠40只,随机分为哮喘组(A组)、生理盐水对照组(C组)、地塞米松治疗组(D组)和罗红霉素治疗组(R组),每组10只.利用卵白蛋白(ovalbumin,OVA)/Al(OH).致敏与OVA雾化吸人激发建立大鼠哮喘模型.免疫组织化学测定肺组织中支气管上皮下MF的一平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达含量,并使用图象分析技术进行积分光密度(integral optical density,IOD)定量分析测定.光镜观察肺组织病理结构变化,图像分析软件分析,并测定肺内支气管总管壁厚度、内管壁厚度、平滑肌层厚度等指标.结果 免疫组织化学和图像分析结果:A组与C组相比,总管壁厚度、内管壁厚度、平滑肌层厚度显著增厚(P<0.01).D组和R组中的内管壁厚度、平滑肌层厚度与A组相比.均显著变薄(P<0.01).R组的总管壁厚度、内管壁厚度、平滑肌层厚度与D组相比差异无统计学意义.定量分析测定的IOD值显示A组支气管上皮下MF α-SMA表达含量较C组显著增加(P<0.01),D组和R组表达含量较A组均减少(P<0.05),R组表达含量与D组相比差异无统计学意义.相关分析结果:支气管上皮下MF α-SMA表达含量(用IOD值表示)和内管壁厚度呈正相关(r=0.913,P<0.01,n=40),和平滑肌层厚度也呈正相关(r=0.626.P<0.01,n=40).结论 MF在气道重塑形成中起重要作用.罗红霉索和地塞米松均可能通过抑制MF增殖和表达起到抗哮喘气道重塑作用.     

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.     

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.