促红素衍生肽对刀豆蛋白A诱导的小鼠急性肝损伤的保护作用

目的探讨外源性给予人工合成的促红素衍生肽(HBSP)对刀豆蛋白A(Con A)诱导的急性免疫性肝损伤的保护作用。方法采用小鼠尾静脉注射Con A建立急性肝损伤模型,造模后2 h给予HBSP治疗(采用尾静脉注射,8nmol/kg,每6小时1次),造模后6、12、24 h分别测定小鼠血清中ALT、AST水平,并进行肝组织病理学检查评估肝损伤程度。测定造模后12 h肝组织中IL-6、IL-1β、TNF-α、IFN-γ、IL-12、IL-4、IL-10、TGF-β表达水平,通过TUNEL及免疫组化检测(分裂型Caspase-3)评估肝脏内肝细胞凋亡情况。结果在Con A诱导的急性肝损伤小鼠模型中,HBSP给药组血清ALT和AST水平显著低于Con A模型组,并且小鼠肝组织炎症坏死程度明显低于Con A模型组。同时,80 nmol/kg的HBSP给药显著提高了注射致死剂量Con A小鼠的生存率。此外,HBSP在mRNA水平抑制肝脏中促炎因子(IL-6、IL-1β、TNF-α、IFN-γ、IL-12)的表达,促进抑炎因子(IL-4、IL-10)的表达,并显著抑制Con A诱导的急性肝损伤相关的细胞凋亡。结论 HBSP对Con A诱导的急性肝损伤具有较好的保护作用,其机制可能与其抑制炎性反应及肝细胞凋亡有关。     

甘草酸铵通过抑制HMGB1的表达减轻ConA诱导的免疫性肝损伤

目的探讨甘草酸铵对刀豆蛋白A(Concanavalin A,ConA)诱导的免疫性肝损伤的保护作用及其可能的分子机制。方法在小鼠尾静脉注射ConA(20 mg/kg)前30 min,腹腔注射甘草酸铵(100 mg/kg)。ConA注射后8 h,检测血清转氨酶水平及肝组织学以评估肝损伤程度,应用酶联免疫法(ELISA)检测肝组织肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)、白介素-10(IL-10)水平。同时,分别采用实时荧光定量PCR和免疫组化检测肝组织高迁移率族蛋白-1(HMGB1)mRNA与蛋白的表达。结果预防性应用甘草酸铵有效地保护ConA诱导的小鼠免疫性肝损伤,表现为血清转氨酶水平与肝脏组织炎症坏死程度明显减轻;与ConA模型组相比,甘草酸铵对肝脏内的促炎症细胞因子TNF-α和IFN-γ并无影响,但增加了抗炎症因子IL-10水平(P<0.01);同时,甘草酸铵预防组肝组织HMBG1基因与蛋白的表达均明显低于模型组(P均<0.01)。结论甘草酸铵有效改善ConA诱导的小鼠免疫性肝损伤,其发挥作用的分子机制与抑制HMBG1表达有关。     

骨髓间充质干细胞治疗ConA诱导的小鼠急性肝损伤的作用及机制

目的:观察骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)在刀豆蛋白A(concanavalin A,Con A)诱导小鼠急性肝损伤中的治疗效果,并对其可能的治疗机制进行探讨.方法:将C57BL/6小鼠的股骨骨髓进行分离、培养、鉴定并体外标记获得BMMSCs;小鼠尾静脉注射不同浓度Con A,选择最适剂量建立急性肝损伤模型;小鼠模型分别经尾静脉注射移植不同剂量(1×105、5×105、1×106、1×107)的氯甲基苯甲酰胺(CM-Dil)标记的BMMSCs,对照组给予等量磷酸盐缓冲液.治疗后24 h检测血清样本中谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)的水平,酶联免疫吸附分析法检测血清中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、干扰素γ(interferon-γ,IFN-γ)及白介素-4(interleukin-4,IL-4)的水平,取肝脏的病理,荧光显微镜下观察BMMSCs在肝组织内的定植情况,并进行肝脏炎症活动度Knodell评分.结果:小鼠BMMSCs体外培养及鉴定后符合BMMSCs的特征,CM-Di L体外标记率可达90%以上;随着Con A注射剂量的增加,小鼠肝组织损伤严重程度增加,15 mg/kg是诱导C57BL/6小鼠急性肝损伤的最合适剂量;Ⅱ组、Ⅲ组小鼠血清ALT、AST、T N F-α、I F N-γ、I L-4的水平以及肝脏Knodell评分均明显低于对照组(P0.05).肝组织内可见到植入的被标记的BMMSCs,而在心、脾、肺等重要脏器并未观察到标记的BMMSCs.结论:BMMSCs可治疗Con A诱导的小鼠急性肝损伤,其机制可能是通过降低TNF-α、IFN-γ及IL-4而实现.     

EGCG对刀豆蛋白A诱导肝损伤小鼠CXCR3表达的影响

目的观察EGCG对刀豆蛋白A(Con A)诱导的肝损伤小鼠肝组织中CXCR3表达的影响。方法 C57BL/6小鼠分成4组:正常对照组、表没食子儿茶素酸酯(EGCG)对照组、Con A模型组,EGCG+Con A模型组。EGCG对照组及EGCG+Con A模型组小鼠造模前给予EGCG口服(5 mg/kg),10 d后两组模型组小鼠通过静脉注射Con A(15 mg/kg)建造肝损伤模型,采血及留取肝组织,HE法检测肝组织病理变化,ELISA法检测肿瘤坏死因子(TNF)α,干扰素(IFN)-γ及CXCR3的水平,免疫组化法检测肝组织中CXCR3的表达。结果 Con A模型组小鼠肝损伤明显,TNF-α、IFN-γ及CXCR3的表达明显增多,与其他组比较有差异显著(P0.05);EGCG干预的模型组肝损伤减轻伴TNF-α、IFN-γ及CXCR3的表达下降,与Con A模型组比较差异明显(P0.05)。免疫组化结果同样显示Con A模型组CXCR3的表达明显增多,EGCG治疗后表达减少。结论 EGCG对Con A诱导的免疫性肝损伤小鼠有保护作用,其机制可能与调节CXCR3的表达有关。     

海珠益肝加味方对免疫性肝损伤小鼠细胞因子的影响

目的:观察海珠益肝加味方对刀豆球蛋白A(Con A)诱导建立的免疫性肝损伤小鼠细胞因子IL-4、IL-6、INF-γ、TNF-a的影响,探讨其作用机制。方法:将40只昆明小鼠随机分为正常组、模型组、醋酸强的松组、海珠益肝加味方组,采用尾静脉注射Con A建立免疫性肝损伤小鼠模型,眼球取血,检测小鼠血清IL-4、IL-6、IFN-γ、TNF-α的水平。结果:与正常组比较,模型组小鼠外周血IL-4、IL-6水平明显降低(P<0.01),IFN-γ、TNF-α水平显著升高(P<0.05);与模型组比较,醋酸强的松组、海珠益肝加味方组小鼠外周血IL-4、IL-6的水平明显升高(P<0.01),IFN-γ、TNF-α水平显著降低(P<0.05)。结论:海珠益肝加味方能升高免疫性肝损伤小鼠细胞因子IL-6、IL-4的分泌水平,降低IFN-γ、TNF-α的水平,恢复Th1/Th2细胞之间的平衡,具有保护肝细胞的作用,可能是其作用机制之一。     

夏枯草硫酸多糖对异烟肼所致小鼠抗结核药物性肝损伤的保护作用及机制

[目的]分析夏枯草硫酸多糖对异烟肼所致小鼠抗结核药物性肝损伤的保护作用,并探究其可能的机制。[方法]将24只SPF级雄性C57 BL/6小鼠随机分为正常对照组、模型组和实验组,每组8只。模型组和实验组每日定时予以异烟肼100 mg/kg灌胃1次。实验组于异烟肼灌胃前予以夏枯草硫酸多糖100 mg/kg灌胃预处理,每4 d灌胃1次,共3次。异烟肼处理2周后处死小鼠,检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活性;肝脏匀浆后检测肝组织丙二醛(MDA)、超氧化物歧化酶(SOD)活性;苏木精-伊红染色观察肝脏组织病理形态学的改变;RT-qPCR技术检测肝脏组织白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)mRNA的表达;Western blot技术检测肝脏组织IL-6和TNF-α蛋白的表达。[结果]与正常对照组相比,模型组和实验组小鼠血清AST、ALT活性水平和肝组织MDA含量升高(P0.001),肝组织SOD活性降低(P0.001),IL-6和TNF-αmRNA和蛋白表达明显升高(P0.001),肝组织病理可见不同程度的肝细胞变性坏死及炎细胞浸润。实验组与模型组相比,小鼠血清AST、ALT活性水平和肝组织MDA含量降低,肝组织SOD活性升高(P0.001),IL-6和TNF-αmRNA和蛋白表达明显降低(P0.001),肝组织病理改变明显减轻。[结论]夏枯草硫酸多糖对异烟肼所致小鼠抗结核药物性肝损伤具有保护作用,其作用机制可能与抗氧化应激反应、降低炎症反应有关。     

肝脏X受体激活对α-GalCer诱导的小鼠肝损伤保护机制探讨

肝脏X受体（LXR）属于核受体超家族成员,对脂类代谢相关基因的转录调控起关键作用,同时具有调节免疫反应和抗炎效应。目的：研究LXR激活对仅．GalCer诱导的小鼠肝损伤保护作用的可能机制。方法：15只C57BL／6J小鼠随机分为正常对照组、α-GalCer模型组和LXR治疗组,后两组以仅α-GalCer腹腔注射诱导肝损伤模型．LXR治疗组于造模前连续7d腹腔注射LXR激动剂T0901317。造模6h后处死小鼠,行肝组织病理学检查和血清AIJT、AST水平检测,免疫组化染色检测肝组织白细胞介素-6（IL-6）表达．蛋白质印迹法检测肝内P13K／Akt／NF—κB信号通路激活情况,实时RT-PCR检测肝组织肿瘤坏死因子-α（TNF-α）、诱导型一氧化氮合酶（iNOS）mRNA表达。结果：与正常对照组相比,α-GalCer模型组小鼠肝损伤明显。血清转氨酶水平升高,肝组织IL-6、TNF-α仅、iNOS表达上调．P13K／Akt／NF—κB信号通路激活。LXR治疗组肝损伤和血清转氨酶水平较α-GalCer模型组显著改善,肝组织炎症介质表达下调,P13K／Akt／NF—κB信号通路激活受抑。结论：LXR激活可调节免疫反应,抑制肝脏炎症,从而显著减轻α-GalCer诱导的小鼠肝损伤．其机制可能与抑制P13K／Akt／NF—κB信号通路激活有关。     

海珠益肝加味方对免疫性肝损伤小鼠的防护作用

目的:观察海珠益肝加味方对刀豆球蛋白A(Con A)诱导建立的免疫性肝损伤小鼠的防护作用及作用机制。方法:将40只昆明小鼠随机分为正常组、模型组、醋酸强的松组、海珠益肝加味方组,采用尾静脉注射Con A建立免疫性肝损伤小鼠模型。造模给药8h后眼球取血,检测小鼠血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、白细胞介素-4(IL-4)和白细胞介素-6(IL-6)水平。摘取肝脏,光镜下观察各组肝组织的病理变化,检测肝组织超氧化物歧化酶(SOD)活性及丙二醛(MDA)的水平,测定肝组织NF-κB-p65表达水平。结果:与模型组比较,对照组和海珠益肝加味方组降低血清ALT、AST水平及肝组织MDA水平,降低肝组织NF-κB-p65表达水平,提高血清IL-4、IL-6水平及其肝组织SOD活性,同时海珠益肝加味方组肝脏病理变化程度明显减轻。结论:海珠益肝加味方能有效降低免疫性肝损伤小鼠转氨酶水平,减轻肝细胞膜的脂质过氧化反应及其肝脏炎症,调控免疫反应的NF-κB通路,具有保护肝细胞的作用。     

异甘草酸镁对刀豆蛋白A诱导的急性肝衰竭小鼠模型的影响

目的研究异甘草酸镁(Mg IG)对刀豆蛋白A(Con A)诱导的急性肝衰竭小鼠模型的作用及机制。方法选取健康BALB/C小鼠90只,随机分为4组,分别为对照组、Mg IG对照组,每组各10只; Con A模型组、Mg IG+Con A预处理组,每组各35只(其中15只用来计算总生存率)。提取小鼠肝脏及外周血,测定生存率、血清转氨酶水平,肝组织HE染色评价小鼠肝损伤程度,TUNEL荧光染色及caspase-3蛋白活性检测评价小鼠肝细胞凋亡水平,液相芯片法检测小鼠血清炎症细胞因子IL-1β和TNFα含量。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 Con A模型组小鼠总生存率为40%,Mg IG+Con A预处理组小鼠总生存率为80%。Con A模型组血清ALT与AST水平较对照组均显著上升(P值均0. 01),Mg IG+Con A预处理组血清ALT与AST水平较Con A模型组均显著降低(P值均0. 01)。对照组、Mg IG对照组、Con A模型组及Mg IG+Con A预处理组病理评分分别为1. 0±0. 2、1. 2±0. 3、3. 7±0. 6、2. 3±0. 5,组间比较差异有统计学意义(F=2. 7,P 0. 05);凋亡细胞/100细胞数值分别为0. 2±0. 1、0. 1±0. 1、7. 8±1. 3、2. 2±0. 4,组间比较差异有统计学意义(F=27. 6,P 0. 001); caspase-3活性分别为0. 813±0. 022、0. 930±0. 033、1. 347±0. 042、1. 060±0. 053,组间比较差异有统计学意义(F=51. 072,P 0. 001)。Con A模型组与对照组比较,血清IL-1β与TNFα水平均显著上升(P值均0. 05)。Mg IG+Con A预处理组与Con A模型组比较,血清IL-1β与TNFα均显著下降(P值均0. 05)。结论异甘草酸镁对刀豆蛋白A诱导的小鼠急性肝衰竭具有明显保护作用,减少肝细胞凋亡并减轻IL-1β和TNFα引起的肝脏炎症反应可能是其缓解肝损伤的机制。     

大麻二酚对刀豆蛋白A诱导小鼠急性免疫性肝炎的保护作用

目的 探讨大麻二酚(cannabidiol, CBD)对刀豆蛋白A(concanavalin A, ConA)诱导小鼠急性免疫性肝炎的保护作用及其机制。方法 选取C57BL/6小鼠30只随机分为正常对照组、ConA模型组、CBD实验组(分为低剂量5 mg/kg、中剂量15 mg/kg、高剂量30 mg/kg),每组6只。实验结束后取小鼠外周血测定血清转氨酶ALT水平，HE染色评估肝脏损伤程度，ELISA法测定肝组织TNF-α、IL-6和IL-10含量，免疫组化及qRT-PCR法检测肝内趋化因子MIP-1α和CXCL10的表达情况。结果 ConA模型组小鼠血清ALT水平较正常对照组明显升高(P<0.01),与ConA模型组相比，不同剂量CBD实验组均能显著降低血清ALT水平(P均<0.01),中、高剂量组ALT水平下降程度较低剂量组更显著。HE染色显示，ConA模型组小鼠肝脏明显坏死、大量炎症细胞浸润，而CBD实验组小鼠肝脏的坏死与炎症则有所不同程度的减轻。ConA模型组小鼠肝脏内细胞因子TNF-α、IL-6水平均较正常对照组明显升高(P<0.01),IL-10则有所降...     

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.