大承气汤对心脏骤停后综合征兔心肺组织蛋白酶激活受体1活性的抑制作用

目的:探讨大承气汤对心脏骤停后综合征(PCAS)兔心、肺组织蛋白酶激活受体1(PAR-1)激活的影响。方法:采用窒息性心脏骤停制备兔PCAS模型,复苏成功兔随机分为PCAS组、大承气汤组,另选5只健康兔作为正常对照组。在ROSC后15 min,大承气汤组给予大承气汤15 g/(kg·d)灌胃。48 h取动物血液检测,后处死动物,取心肺组织,采用Western blot测定PAR-1蛋白含量。结果:PCAS组兔心肺组织中PAR-1蛋白含量降低(均P0.01)。大承气汤组心肺组织中PAR-1蛋白含量较PCAS组升高(P0.05)。结论:PCAS发生时,心肺组织存在PAR-1过度激活,大承气汤对PCAS兔PAR1激活有抑制作用。     

人腰椎间盘细胞凋亡途径的探讨

取30例腰椎间盘突出症患者的腰椎间盘髓核组织标本（观察组）和10例创伤性椎间盘损伤患者的椎间盘髓核组织标本（对照组）,应用免疫组织化学方法 测定标本中半胱天冬酶-3（caspase-3）、半胱天冬酶-8（caspase-8）、半胱天冬酶-9（caspase-9）和细胞色素C的阳性表达情况;电镜下观察凋亡细胞的形态学变化。结果 观察组caspase-3、caspase-9、细胞色素C阳性细胞表达率明显高于对照组（P〈0.001）;caspase-8表达率较低,两组比较无统计学意义;电镜下观察组出现典型的凋亡细胞形态学变化,细胞凋亡数目明显高于对照组。认为人腰椎间盘髓核细胞通过线粒体依赖途径发生细胞凋亡。     

缺氧后适应与心肌营养素-1对心肌细胞缺氧复氧的保护作用及机制探讨

目的 探讨缺血后适应与心肌营养素-1（CT-1）对心肌细胞缺氧复氧的保护作用及其机制.方法 将体外培养的H9C2细胞株分为正常对照组（a组）、缺氧复氧组（b组）、缺氧复氧＋缺氧后适应组（c组）、缺氧复氧＋缺氧后适应＋ CT-1组（d组）、缺氧复氧＋缺氧后适应＋CT-1＋ ERK抑制剂组（e组）、缺氧复氧＋缺氧后适应＋CT-1＋二甲基亚砜组（f组）,MTS法检测各组细胞存活率、流式细胞仪检测细胞凋亡率、Western blot检测细胞外调节激酶（ERK1/2）和磷酸化ERK1/2（p-ERK1/2）的表达、Q-PCR检测Bad mRNA的表达.结果 与a组比较,其他各组细胞存活率下降,细胞凋亡率、Bad mRNA和p-ERK1/2（除外e组）表达量增加,P均＜0.05;与b组比较,c组细胞存活率、p-ERK1/2表达量均增加,Bad mRNA表达、凋亡率下降,P均＜0.05;与c组比较,d组细胞存活率、p-ERK1/2表达量均增加,细胞Bad mRNA表达、凋亡率下降（P均＜0.05）;与d组比较,e组细胞存活率下降,细胞凋亡率、Bad mRNA表达量增加,p-ERK1/2表达量下降;d、e组各项指标指标比较差异无统计学意义.结论 缺氧后适应可减少心肌细胞缺氧复氧损伤,联合CT-1后保护作用明显加强,而ERK1/2抑制剂可阻断这种保护作用;该保护机制可能与CT-I激活ERK1/2信号通路,下调Bad mRNA的表达有关.     

组织金属蛋白酶抑制因子3重组蛋白诱导小鼠成骨细胞MC3T3-E1凋亡

目的 研究组织金属蛋白酶抑制因子3（TIMP-3）重组蛋白对MC3T3-E1成骨细胞凋亡的影响。方法 细胞存活率和凋亡分别用MTT及ELISA检测。Fas，FasL，Bcl-2，Bax，caspase-3，caspase-8，细胞色素c的表达以及JNK，p38，细胞外信号调节激酶（ERK）1／2的磷酸化水平用Western印迹检测。结果 MTT及ELISA检测提示TIMP-3降低MC3T3-E1细胞存活率，诱导MC3T3-E1细胞凋亡。TIMP-1增加Fas、FasL蛋白表达，对Bax、Bcl-2蛋白表达无影响，但可诱导细胞色素c的释放及caspase-8、caspase-3的活化。TIMP-3促进p38和ERK磷酸化，而PD098059（ERK阻断剂）和SB203580（p38阻断剂）消除p38和ERK的促凋亡作用。结论 TIMP-3诱导MC3T3-E1细胞凋亡的信号途径为凋亡受体Fas介导，并有p38、ERK信号转导途径参与。     

鼻咽癌组织中 ERK1／2、p-ERK1／2和 MMP-9蛋白的表达变化及其相关性

目的：观察细胞外调节蛋白激酶1/2（ERK1/2）、磷酸化细胞外调节蛋白激酶1/2（p-ERK1/2）和基质金属蛋白酶9（MMP-9）在鼻咽癌（NPC）组织中的表达及相关性,并探讨其意义。方法采用免疫组织化学染色SP法检测75份NPC组织（ NPC组）和27份鼻咽黏膜慢性炎组织（炎性组）中ERK1/2、p-ERK1/2和MMP-9蛋白的表达,分析三者在NPC组织中表达的相关性。结果 NPC组ERK1/2、p-ERK1/2和MMP-9蛋白阳性表达率均显著高于炎性组（P均＜0．01）,χ2值依次为19．8、17．1、21．3;NPC组织中ERK1/2、p-ERK1/2和MMP-9蛋白三者间的表达呈正相关（P均＜0．01）,其中ERK1/2与p-ERK1/2、MMP-9蛋白的r值分别为0．58、0．34,p-ERK1/2、MMP-9蛋白的r值为0．34。结论 ERK1/2、p-ERK1/2和MMP-9蛋白在NPC组织中呈高表达且三者间呈正相关,p-ERK1/2可能通过调节MMP-9表达参与NPC侵袭、转移。     

愈肾合剂对糖尿病肾病大鼠肾脏细胞凋亡及ERK信号转导系统的影响

目的观察愈肾合剂对糖尿病(DN)肾病大鼠肾脏细胞凋亡及ERK信号转导系统的影响。方法 SD大鼠单侧肾切除加腹腔注射链脲佐菌素(STZ)复制DN模型,每日愈肾合剂灌胃,8 w后流式细胞术检测肾脏皮质细胞凋亡数目及caspase-9和caspase-3蛋白表达,采用Western印迹方法检测肾脏皮质细胞内磷酸化ERK1/2蛋白质的表达。结果愈肾合剂明显减少DN大鼠肾脏皮质细胞凋亡数目,降低caspase-9和caspase-3的蛋白表达,激活ERK信号通路。结论愈肾合剂可通过抗凋亡途径发挥肾脏保护作用。     

氧化苦参碱对喉癌细胞株Hep-2抑制增殖及诱导凋亡的实验研究

目的观察氧化苦参碱（Oxy）对喉癌细胞株（Hep-2）的抑制增殖及诱导凋亡作用。方法培养喉癌细胞株（Hep-2）,MTT比色法及流式细胞术测定Oxy对喉癌细胞株Hep-2抑制增殖及诱导凋亡的作用。免疫沉淀法纯化蛋白并ERK1／2试剂盒测定细胞外调节信号激酶（ERK）活性;Western blot测定p-ERK、Bax及Bcl-2表达。结果Oxy明显抑制Hep-2细胞增殖,诱导Hep-2细胞凋亡;Oxy可抑制ERK活性及p-ERK表达,同时上调Bax表达,降低Bcl-2表达。结论Oxy可以抑制Hep-2增殖,机制与抑制ERK活性及p-ERK表达有关;同时可诱导凋亡,其途径与上调Bax表达,降低Bcl-2表达有关。     

神经调节蛋白1通过ERK1-2通路减轻心肌细胞缺氧复氧损伤

目的探究神经调节蛋白1(NRG-1)通过细胞外调节蛋白激酶1/2(ERK1/2)通路减轻心肌细胞缺氧复氧(H/R)损伤的作用。方法培养心肌H9c2细胞株,随机分为常规条件下用不含药物DMEM处理的对照组、H/R条件下用不含药物DMEM处理的H/R组、H/R条件下用含NRG-1 DMEM处理的NRG-1组、H/R条件下用含NRG-1及ERK1/2抑制剂PD98059处理的NRG-1+PD组。检测细胞增殖活力、凋亡率及细胞中的凋亡基因、炎症指标、ERK1/2。结果 H/R组细胞的OD_(490)水平明显低于对照组,细胞凋亡率、细胞中cleaved Caspase-8、cleaved Caspase-9、cleaved Caspase-3、核因子κB(NF-κB)、ERK1/2的表达水平及培养基中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、γ干扰素(IFN-γ)的含量明显高于对照组;NRG-1组细胞的OD_(490)水平及细胞中ERK1/2的表达水平明显高于H/R组,细胞凋亡率、细胞中cleaved Caspase-8、cleaved Caspase-9、cleaved Caspase-3、NF-κB的表达水平及培养基中TNF-α、IL-1β、IFN-γ的含量明显低于H/R组;NRG-1+PD组细胞的OD_(490)水平及细胞中ERK1/2的表达水平明显低于NRG-1组,细胞凋亡率、细胞中cleaved Caspase-8、cleaved Caspase-9、cleaved Caspase-3、NF-κB的表达水平及培养基中TNF-α、IL-1β、IFN-γ的含量明显高于NRG-1组。结论 NRG-1通过激活ERK1/2通路减轻心肌细胞H/R损伤。     

雷公藤多甙对NOD小鼠胰岛β细胞半胱天冬蛋白酶表达的影响

用环磷酰胺促进NOD小鼠1型糖尿病的发病，腹腔注射雷公藤多甙（TWP）4周，监测糖尿病发病率，TUNEL法检测胰岛细胞凋亡，免疫组化法观察胰岛β细胞半胱天冬蛋白酶（caspase）3的表达，RT-PCR方法检测胰腺caspase-8 mRNA的表达。结果提示TWP组糖尿病发病率下降，胰岛β细胞凋亡减少，胰岛β细胞caspase-3蛋白及胰腺caspase-8 mRNA的表达均减少。     

抗氧化剂和内质网钙释放阻滞剂对老年大鼠肾脏缺血/再灌注损伤中凋亡的影响

目的 探讨联合应用抗氧化剂BHA（Butylated Hydroxyanisole,叔丁基羟基茴香醚）和内质网钙释放阻滞剂TMB-8[8-（N,N-diethylamino）-noctyl-3,4,5-trimethoxybenzoate）]对老年大鼠肾脏缺血/再灌注（I/R）损伤时肾组织中细胞凋亡蛋白酶-3（caspase-3）表达及活性的影响.方法 27月龄Wistar大鼠分别随机分为假手术组、I/R模型组、I/R＋BHA＋TMB-8组,双侧肾动脉夹闭30 min/再灌注18 h制作模型.观察肾功能改变,原位末端标记（TUNEL）法检测肾小管上皮细胞凋亡情况,Northern blot检测肾组织中caspase-3 mRNA表达,比色法测定肾组织中caspase-3活性.结果 肾脏I/R损伤时,老年大鼠肾功能明显减退,大量肾小管上皮细胞凋亡,肾组织caspase-3 mRNA表达及活性增加;BHA＋TMB-8能显著改善老年大鼠肾功能,减轻肾小管上皮细胞凋亡,降低肾组织caspase-3表达及活性.结论 BHA＋TMB-8联合治疗能明显抑制老年大鼠肾脏I/R损伤时肾组织caspase-3表达及活性,从而减轻肾小管上皮细胞凋亡,保护肾功能.     

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.