软脉降脂胶囊对载脂蛋白E基因敲除小鼠动脉粥样硬化影响的实验研究

目的观察软脉降脂胶囊对载脂蛋白E基因敲除载脂蛋白E基因敲除小鼠(ApoE)高脂血症及动脉粥样硬化(atherosclerosis,AS)的影响.方法6周龄ApoE小鼠,随机分为高脂血症组与软脉降脂胶囊大、中、小剂量组,相同遗传背景的同龄正常C57BL/6J小鼠作为正常对照组.软脉降脂胶囊组灌服其生药,高脂血症组、正常对照组均灌服生理盐水.连续灌胃24周后,下腔静脉取血测血脂;取主动脉做形态学观察及图像分析.结果高脂血症组血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白(LDL-C)明显高于正常对照组(P＜0.05),软脉降脂胶囊组明显低于高脂血症组(P＜0.05);图像分析测量结果显示高脂血症组AS病变显著,与正常对照组比较斑块总面积明显增加(P＜0.01);软脉降脂胶囊组AS病变较高脂血症纽明显减轻,斑块总面积明显减小(P＜0.01).结论软脉降脂胶囊可降低ApoE小鼠血清TC、TG、LDL-C含量,有对抗动脉粥样硬化斑块形成的作用.     

载脂蛋白E基因敲除小鼠不同周龄动脉粥样硬化的病理变化

目的研究载脂蛋白E基因敲除小鼠不同时间段的动脉粥样硬化的病理进展，探讨不同饮食对载脂蛋白E基因敲除小鼠动脉粥样硬化进展的影响。方法采用6周龄40只载脂蛋白E基因敲除小鼠，20只给予高脂膳食饲料，20只给予普通饮食，分别于15周龄、19周龄、24周龄、28周龄、36周龄5个时间段各取4只麻醉处死做病理检测。结果随着时间进展，载脂蛋白E基因敲除小鼠动脉粥样硬化程度逐渐加重，经历了从脂质条纹到粥样斑块形成典型的动脉粥样硬化病理过程；高脂饮食组比普通饮食组的载脂蛋白E基因敲除小鼠动脉粥样硬化病理改变严重。结论载脂蛋白E基因敲除小鼠是研究动脉粥样硬化的可靠模型，高脂饮食可以加速载脂蛋白E基因敲除小鼠动脉粥样硬化的病理进程。     

鸢尾素对载脂蛋白E基因敲除小鼠动脉粥样硬化的影响

目的研究鸢尾素对载脂蛋白E基因敲除(ApoE~(-/-))小鼠动脉粥样硬化的影响。方法将ApoE~(-/-)小鼠随机分为正常对照组、动脉粥样硬化模型组、鸢尾素组,每组10只,分别给予普通饮食+生理盐水、高脂饮食+生理盐水、高脂饮食+鸢尾素,饲养至12周后处死,处死前测体重,用化学法测定总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDLC)、高密度脂蛋白胆固醇(HDLC)、血清超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量。采用HE染色和整体油红O染色分析主动脉粥样斑块面积大小,实时荧光定量(qRT-PCR)检测主动脉CD36、NF-κB p65的mRNA表达,Western blot检测主动脉CD36、NF-κB p65、SOD的蛋白表达。结果动脉粥样硬化模型组主动脉斑块面积明显高于正常对照组,内膜明显增厚,而鸢尾素组较动脉粥样硬化模型组斑块面积减少,内膜增厚程度下降。与正常对照组比较,动脉粥样硬化模型组小鼠体重、TC、TG、LDLC、MDA水平升高,NF-κB p65、CD36mRNA和蛋白表达增加,而血清SOD活性、HDLC水平减低(P0.05)。与动脉粥样硬化模型组比较,鸢尾素组小鼠体重、TC、TG、LDLC、MDA水平降低,NF-κB p65、CD36 mRNA和蛋白表达下调,而血清SOD活性、HDLC水平升高(P0.05)。结论鸢尾素能减轻ApoE~(-/-)小鼠动脉粥样硬化的发生发展,这可能与其降低小鼠体重和血脂、抑制炎症反应、抗氧化应激有关。     

CCR2受体介导单核细胞趋化蛋白1诱导的人脐静脉内皮细胞凋亡

目的观察载脂蛋白E基因敲除小鼠头臂干动脉粥样硬化斑块在进展过程中斑块内部不同组分的变化情况。方法将18只8周龄载脂蛋白E基因敲除(ApoE-/-)小鼠随机分为三组,每组6只,给与西方膳食饲料分别饲养至24周龄、32周龄和40周龄后,麻醉开胸经下腔静脉取血后采用生物化学方法检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDLC)和高密度脂蛋白胆固醇(HDLC);取出头臂干动脉后用4%多聚甲醛固定,常规脱水,石蜡包埋并切片得到....     

载脂蛋白E基因敲除小鼠动脉粥样硬化斑块中血小板因子4和Toll样受体2的表达

目的观察高脂饲养的载脂蛋白E基因敲除小鼠动脉粥样硬化斑块表达Toll样受体2和血小板因子4的情况,探讨血小板因子4对内皮细胞Toll样受体2表达的影响。方法高脂饲料喂养载脂蛋白E基因敲除小鼠12周,建立动脉粥样硬化模型。安乐死处死动物,原位灌流固定,取主动脉于10%中性缓冲福尔马林中固定,石蜡包埋连续切片,HE染色观察动脉粥样硬化斑块形态,免疫组织化学检测斑块中Toll样受体2和血小板因子4的表达。结果载脂蛋白E基因敲除小鼠血脂水平明显增高,主动脉HE染色可见态动脉粥样硬化病变。在载脂蛋白E基因敲除小鼠主动脉富含脂质斑块中Toll样受体2表达上调,其中血管内皮细胞、巨噬细胞表达Toll样受体2明显增多。载脂蛋白E基因敲除小鼠主动脉斑块中也发现有血小板因子4表达,主要在内皮细胞和动脉粥样硬化斑块肩部。结论1.载脂蛋白E基因敲除小鼠粥样斑块中Toll样受体2表达上调,并且Toll样受体2主要表达在粥样斑块的内皮细胞和巨噬细胞上。2.载脂蛋白E基因敲除小鼠动脉粥样硬化斑块中可见血小板因子4表达。     

中西药物对载脂蛋白E基因敲除小鼠动脉粥样硬化的干预作用

美国洛克菲勒大学生化遗传与代谢实验室和北卡罗莱那大学病理遗传实验室应用胚胎干细胞基因敲除 ( geneknockout)技术 ,于 1992年培育成功载脂蛋白E (apolipoproteinE ,ApoE)基因敲除小鼠 ,亦称ApoE基因缺陷小鼠或ApoE基因摧毁小鼠(本文中统称为ApoE基因敲除小鼠 )。ApoE是清除乳糜微粒和极低密度脂蛋白的受体的配体 ,因此 ,缺乏ApoE则会导致血液循环中富含胆固醇的物质积累而更加容易引起动脉粥样硬化(atherosclerosis ,AS)病灶形成。ApoE基因敲除小鼠无论正常或高脂饮食均可形成严重的高脂血症及AS病灶[1,2 ] ,成为高脂血症及AS…     

吡格列酮抑制载脂蛋白E基因敲除小鼠主动脉粥样硬化病变进展及可能机制

目的观察吡格列酮对高脂饲料喂养的载脂蛋白E基因敲除小鼠主动脉粥样硬化病变及其血浆脂联素水平的影响,以探讨噻唑烷二酮类药物抗动脉粥样硬化作用及其可能的机制。方法取9只17周龄雄性C57BL/6J小鼠普通饲料喂养4周为对照组;另取31只同周龄雄性载脂蛋白E基因敲除小鼠随机分成三组给予高脂饲料喂养4周,其中模型组(n=10)单用高脂饲料喂养,低剂量治疗组(n=10)经胃管给予吡格列酮10 mg/(kg.d),高剂量治疗组(n=11)经胃管给予吡格列酮20 mg/(kg.d)。所有试验小鼠均观察其主动脉粥样硬化病变及血脂、血浆脂联素水平。结果与对照组比,所有载脂蛋白E基因敲除小鼠血脂水平显著升高并伴有主动脉内膜明显增厚及内膜下粥样斑块形成;与对照组(18.96±4.89μg/L)比,模型组血浆脂联素水平(12.41±3.84μg/L)显著下降,而两治疗组脂联素水平(18.78±7.24和24.00±4.71μg/L)比模型组显著升高,主动脉粥样硬化病变却减轻,且这种变化在高剂量治疗组更明显。结论吡格列酮抑制载脂蛋白E基因敲除小鼠主动脉粥样硬化病变的进展可能与其提高血浆脂联素水平有关。     

南蛇藤素对载脂蛋白E基因敲除小鼠主动脉壁C反应蛋白及组织因子表达的影响

目的探讨南蛇藤素对高脂饲养载脂蛋白E基因敲除小鼠在动脉粥样硬化病变形成的早期动脉壁内C反应蛋白和组织因子表达的影响。方法8周龄雄性载脂蛋白E基因敲除小鼠12只,随机分为南蛇藤素干预组或二甲基亚砜溶剂对照组,每组各6只。均给予高脂饲养8周,在高脂饲养的后4周,分别给予南蛇藤素2mg/(kg·d)或相当剂量的二甲基亚砜腹腔注射。麻醉处死小鼠后,取小鼠主动脉,以石蜡包埋,行主动脉根部连续切片;以HE染色观察形态学变化,免疫组织化学法检测主动脉壁内C反应蛋白和组织因子的表达水平,以Image Pro Plus6.0软件进行图像分析。结果南蛇藤素干预组主动脉粥样硬化斑块面积明显小于对照组,分别为4947±1277μm2和8403±2535μm2(P<0.05);南蛇藤素组主动脉粥样斑块面积/血管壁面积比值明显小于对照组(P<0.05);南蛇藤素组动脉壁C反应蛋白表达水平较对照组明显减少,平均光密度值分别为0.0152±0.0052与0.0256±0.0026(P<0.05);动脉粥样硬化斑块内组织因子表达水平较对照组明显减少,平均光密度值分别为0.0326±0.0132与0.0763±0.0347(P<0.05)。结论南蛇藤素可能通过抑制载脂蛋白E基因敲除小鼠炎症反应和动脉壁中C反应蛋白的表达而发挥抗动脉粥样的作用;还可能通过减少粥样斑块中组织因子的产生,而进一步起到稳定动脉粥样硬化斑块的作用。     

血管紧张素Ⅱ对载脂蛋白E敲除小鼠主动脉粥样硬化斑块细胞外基质金属蛋白酶诱导因子表达影响

目的探讨血管紧张素Ⅱ对载脂蛋白E敲除小鼠主动脉粥样硬化斑块细胞外基质金属蛋白酶诱导因子表达的影响。方法载脂蛋白E基因敲除小鼠经高脂饮食饲养建立动脉粥样硬化模型,用血管紧张素Ⅱ干预。用免疫组织化学法观察粥样硬化斑块内细胞外基质金属蛋白酶诱导因子表达,用RT-PCR及Western blotting检测主动脉内细胞外基质金属蛋白酶诱导因子表达。结果血管紧张素Ⅱ干预组细胞外基质金属蛋白酶诱导因子在动脉粥样硬化斑块内阳性表达较对照组明显增加;血管紧张素Ⅱ干预组主动脉内细胞外基质金属蛋白酶诱导因子mRNA及蛋白表达较对照组明显增加。结论血管紧张素Ⅱ能诱导主动脉粥样硬化斑块内细胞外基质金属蛋白酶诱导因子的表达。     

苦瓜蛋白对载脂蛋白E基因敲除小鼠动脉粥样硬化形成及小肠胆固醇转运相关基因表达的影响

目的 探讨苦瓜蛋白对载脂蛋白E基因敲除小鼠小肠胆固醇转运相关基因表达的影响,阐明苦瓜蛋白在动脉粥样硬化发生中的作用机制.方法 选用36只6周龄的雄性载脂蛋白E基因敲除小鼠,随机分为普食组(对照组)、高脂高胆固醇饲料组(高脂组)、高脂高胆固醇+苦瓜蛋白组(苦瓜蛋白组).喂养12周后处死,比较各组血脂水平,苏丹Ⅳ染色观察全主动脉粥样硬化病变程度,主动脉窦冰冻切片油红O染色检测脂质沉积,逆转录聚合酶链反应和Western Blot分别检测ATP结合盒转运体G5和G8的mRNA和蛋白表达.结果 高脂组总胆固醇、甘油三酯、低密度脂蛋白胆固醇和高密度脂蛋白胆固醇的水平比对照组明显升高(P＜0.05),而苦瓜蛋白组总胆固醇(14.52±1.06 mmol/L比18.21±1.95 mmol/L)、甘油三酯(0.51±0.11 mmol/L比0.92±0.07 mmol/L)和低密度脂蛋白胆固醇(4.12 mmol/L±0.95比7.16±0.98 mmol/L)水平显著低于高脂组(P＜0.05),高密度脂蛋白胆固醇的水平与高脂组比差异无显著性(P＞0.05).苦瓜蛋白组主动脉斑块面积(31.2%±3.6%比69.6%±4.8%)和主动脉窦脂质面积(26.02±3.07 μ m2/比45.23±11.2 μm2)比高脂组显著减少.苦瓜蛋白组小肠中ATP结合盒转运体G5和G8的mRNA和蛋白表达都比高脂组明显升高(P＜0.05).结论 苦瓜蛋白可通过增加载脂蛋白E基因敲除小鼠小肠中ATP结合盒转运体G5和G8的表达,降低血脂水平,减少主动脉斑块及主动脉窦脂质沉积.     

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.