阿霉素对大鼠心肌诱导型一氧化氮合酶mRNA表达的影响

目的研究阿霉素(ADM)对大鼠心肌诱导型一氧化氮合酶(iNOS)mRNA表达的影响。方法将雄性wistar大鼠24只,随机分为ADM组和对照组,每组12只,ADM组按每次给ADM 2 mg/kg腹腔注射,隔日一次共6次;对照组给等体积的生理盐水腹腔注射。于实验第30 天,应用生化方法测定心肌组织中的一氧化氮(NO)水平及iNOS的活性;用TUNEL法检测心肌细胞的凋亡指数;用RT—PCR方法检测心肌组织iNOS mRNA的表达。结果与对照组比较ADM组大鼠心肌组织NO、iNOS活性增加(P< 0.01),心肌细胞凋亡指数及iNOS mRNA的表达均显著增高(P< 0.01)。结论ADM能诱导大鼠心肌细胞iNOS mRNA表达增加,使NO合成增多,引起细胞凋亡而参与对心肌的损害。     

细胞间黏附分子-1在心肌炎小鼠心肌中的表达及黄芪的干预作用

目的研究细胞间黏附分子-1(ICAM-1)在病毒性心肌炎小鼠心肌组织中的表达,并探讨黄芪对其表达的影响.方法每只BALB/c小鼠腹腔接种半数组织感染率(TCID50)为108/mL的柯萨奇B3病毒0.1 mL.黄芪治疗组于接种病毒后24 h起灌喂黄芪30mg/g,每日1次,共5d.接种病毒后第7天、第14天、第21天分别处死小鼠,免疫组化法检测心肌组织ICAM-1表达.结果心肌炎小鼠心肌细胞和血管内皮细胞ICAM-1表达明显,而正常对照组仅少量血管内皮细胞表达ICAM-1,黄芪治疗可下调ICAM-1表达.结论病毒性心肌炎小鼠心肌组织ICAM-1表达增强,早期黄芪治疗有减弱ICAM-1表达的作用.     

柯萨奇B病毒性心肌炎小鼠急性期心肌组织微小RNA1初始体和连接蛋白43的表达

目的观察柯萨奇B病毒性心肌炎小鼠急性期心肌组织微小RNA1初始体(pri-miRNA-1)和连接蛋白43(Cx43)表达变化,探讨病毒性心肌炎(VMC)室性心律失常发生机制。方法40只4周龄雄性Balb/c小鼠随机分为VMC组(n=20)和对照组(n=20)。VMC组小鼠腹腔注射柯萨奇病毒B3(CVB3)Nancy株悬液0.1ml,对照组小鼠腹腔注射不含病毒的RPMI1640培养基0.1ml,分别于接种病毒后第14天无痛苦处死全部小鼠并留取心脏。逆转录-聚合酶链反应(RT-PCR)检测心室肌组织pri-miRNA-1的表达,免疫组织化学法检测Cx43蛋白水平表达,并进行半定量分析。结果①VMC组小鼠心室肌组织pri-miRNA-1表达量明显高于对照组(0.82±0.04vs0.63±0.07,P(0.01);②VMC组小鼠心室肌组织炎症病灶中变性、坏死周围心肌细胞Cx43表达明显减弱,甚至阴性,分布不规则,Cx43蛋白表达明显低于对照组(0.27±0.01vs0.42±0.02,P(0.01);③VMC组小鼠心室肌组织的pri-miRNA-1表达量与Cx43蛋白水平呈显著负相关(r=-0.868,P(0.01)。结论CVB心肌炎小鼠急性期心肌组织pri-miRNA-1表达上调,Cx43蛋白表达下降,miRNA-1可能通过抑制Cx43表达促进室性心律失常的发生。     

黄芪甲甙对小鼠柯萨奇B3病毒性心肌炎的抗氧化作用

目的观察黄芪甲甙对小鼠柯萨奇B3病毒性心肌炎的抗氧化作用。方法Balb/c小鼠50只随机分5组(每组10只),空白对照组腹腔无菌注射不含病毒的Eagle’s培养基0.1 mL,在腹腔注射病毒培养基30 min后,以生理盐水0.1 mL灌胃,共7d;病毒性心肌炎对照组小鼠每只腹腔注射0.1 mL内含1×102TCID50柯萨奇B3病毒的Eagle’s培养基,在腹腔注射含病毒的Eagle’s培养基30 min后,以生理盐水0.1 mL灌胃,共7 d;黄芪甲甙低、中、高剂量干预组在腹腔注射病毒30 min后,用黄芪甲甙剂量分别为0.07、0.2、0.6 g/(kg.d)0.1 mL灌胃,共7 d。观察小鼠生存数;心肌病变积分;心肌谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活力及逆转录聚合酶链反应检测铜锌超氧化物歧化酶-mRNA水平表达。结果(1)与空白对照组比较,病毒性心肌炎对照组小鼠的生存数明显下降,心肌病变积分明显升高(3.23±0.83,P<0.01);与病毒性心肌炎对照组比较,黄芪甲甙高剂量干预组可明显提高小鼠的生存数(P<0.01),心肌病变积分明显下降(1.86±0.59比3.23±0.83,P<0.01);(2)与病毒性心肌炎对照组比较,黄芪甲甙高剂量干预组心肌组织谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活力呈增高趋势,心肌组织谷胱甘肽过氧化物酶(39.73±8.02比9.99±2.71,P<0.05)、过氧化氢酶(6.35±2.33比1.36±0.87,P<0.05)、超氧化物歧化酶(65.71±4.93比40.15±6.03,P<0.01)活力明显增强,铜—锌超氧化物歧化酶-mRNA水平表达明显增高(0.43±0.11比0.32±0.01,P<0.01)。但黄芪甲甙低、中剂量干预组,虽然也在一定程度使心肌组织谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活力及逆转录聚合酶链反应检测铜—锌超氧化物歧化酶-mRNA表达水平增加,有一定的量效关系,但与病毒性心肌炎对照组比较无统计学意义(P>0.05)。结论黄芪甲甙通过增加心肌抗氧化酶活力,对小鼠柯萨奇B3病毒性心肌炎具有明显的保护作用。     

黄芪对病毒性心肌炎小鼠热休克蛋白70及Bcl-2、Bax基因表达的影响

目的:研究黄芪对病毒性心肌炎(VMC)小鼠热休克蛋白(HSP)70及凋亡相关基因蛋白产物表达的影响。方法:体外培养乳鼠心肌细胞,用Western Blot观察心肌细胞HSP70、Bcl-2及Bax变化;120只Balb/c小鼠随机分为正常对照组、病毒组以及黄芪干预组,采用腹腔接种柯萨奇病毒制做VMC小鼠模型,免疫组化检测各组小鼠Bcl-2和Bax基因表达的情况,并用图像分析系统进行分析。结果:黄芪干预组心肌细胞HSP70及Bcl-2表达明显高于病毒组,2组Bax表达无差异;黄芪干预组心肌组织Bcl-2表达明显高于病毒组,2组Bax表达亦无差异。结论:黄芪可能通过上调心肌细胞Bcl-2、HSP70的表达来抑制病毒所致的心肌细胞凋亡。     

益心饮对感染柯萨奇B3病毒小鼠心肌组织穿孔素和颗粒酶B基因表达的影响

本实验在小鼠急性病毒性心肌炎模型上观察了益心饮对心肌穿孔素和颗粒酶BmRNA表达水平的影响。1 .材料与方法 :雄性Balb/c小鼠、4～ 6周龄、体重 (2 0±2 ) g ,1 2 0只 ,分为正常对照组 ,病毒模型组 ,病毒 黄芪对照组 ,病毒 益心饮低、中、高剂量组 ,每组各 2 0只。除正常对照组外 ,均于实验当日在小鼠腹腔注射柯萨奇B3病毒(CVB3 )稀释液 0 2ml复制病毒性心肌炎动物模型。正常对照组腹腔注射 0 2ml不含病毒的稀释液。实验当日在病毒接种 30min后 ,各组灌胃黄芪注射液 1 0 g·kg-1 ·d-1 、益心饮 2 5g·kg-1 ·d-1 、5g·kg-1 ·d-…     

柯萨奇-腺病毒受体在扩张型心肌病心肌中的表达及意义

目的探讨柯萨奇-腺病毒受体(CAR)在扩张型心肌病(DCM)心肌组织中的表达及其在DCM发病中的意义.方法利用多聚酶链反应(PCR)技术检测20例原位异体心脏移植的DCM受体心肌组织以及5例正常对照心肌组织中柯萨奇B组病毒(CVB)、腺病毒(ADV)基因,采用免疫组化及逆转录多聚酶链反应(RT-PCR)检测心肌柯萨奇-腺病毒受体mRNA及蛋白表达水平.结果20例DCM心肌标本中,柯萨奇B组病毒阳性7例(柯萨奇病毒组),腺病毒阳性5例(腺病毒组);其余8例柯萨奇B组病毒、腺病毒均阴性(其它组),5例正常对照柯萨奇B组病毒、腺病毒也均为阴性.正常对照和其它组心肌中均无柯萨奇-腺病毒受体mRNA及蛋白表达,而在柯萨奇病毒组和腺病毒组其表达水平明显增加,但两组之间比较,差异无显著性.结论柯萨奇-腺病毒受体表达上调可能是导致柯萨奇病毒或腺病毒易感性增加,促使病毒性心肌炎向扩张型心肌病转化的重要原因.     

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     

柯萨奇B3病毒感染小鼠心肌硫氧还蛋白的表达

目的研究硫氧还蛋白(thioredoxin,Trx)在柯萨奇B3病毒感染小鼠心肌炎模型中表达的变化,探讨Trx在病毒性心肌损伤中的作用。方法建立柯萨奇B3病毒感染鼠心肌炎模型,采用光镜检测心肌形态改变,应用RT鄄PCR技术比较接种病毒后不同时间点(7,14,21d)TrxmRNA表达变化。结果7d时心肌病损严重;14d时部分心肌病损开始修复;21d时心肌修复较完全。接种病毒后21d时,硫氧还蛋白mRNA表达与对照组比较明显上调(P<0.01),7d和14d时同对照组相比表达无显著性差异(P>0.05)。结论在急性柯萨奇B3病毒性心肌炎中,Trx可能被急性炎症刺激素特异地诱导,在疾病的后期TrxmRNA表达上调,促进心肌修复,提示适当剂量的Trx可能对急性柯萨奇B3病毒性心肌炎有潜在治疗作用。     

柯萨奇B病毒诱导小鼠心肌病变的实验研究

目的 研究柯萨奇B病毒感染对ICR小鼠心肌组织亲嗜性及损害作用,探讨病毒感染与心肌病变之间的关系.方法 ICR小鼠130只随机分为感染组(120只)和对照组(10只),感染组再分为CVB3组(60只)和cVB4组(60只).CVB3组每只腹腔注射4×100TCID50CVB3/MKP 0.2 ml,CVB4组每只腹腔注射4×100TCID50 CVB4/jlu06 0.2 ml,对照组每只腹腔注射PBS 0.2 ml.分别在感染后1、4、8、15、27和60 d处死小鼠,无菌取心肌组织,收集血液分离血清,用于病毒滴度的测定及病理学分析.结果 两株病毒均在小鼠心肌组织中有较高滴度增殖,并导致了小鼠心肌组织不同程度的损害.结论 CVB3/MKP株具有较强的心肌亲嗜性,可引起ICR小鼠心肌组织的严重损伤,该株病毒感染可能与心肌炎的发生密切相关.     

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.