Compound48/80对载脂蛋白E基因敲除小鼠颈总动脉套环诱导斑块的影响

目的采用肥大细胞脱颗粒剂———Compound 48/80作用于颈总动脉套环的载脂蛋白E基因敲除小鼠,以探讨肥大细胞脱颗粒对动脉粥样硬化斑块的影响及其可能作用机制。方法载脂蛋白E基因敲除小鼠高脂高胆固醇饲料喂养,行右颈总动脉套环术后分为实验组和对照组,分别腹腔注射Compound 48/80或D-hank’s。第4次注射后30 min,安乐死,取材。全自动生物化学分析仪测定血清中脂质含量,比色法测定血清中类胰蛋白酶活性,苏木素—伊红染色观察颈总动脉病理改变,甲苯胺蓝肥大细胞染色,免疫组织化学检测平滑肌肌动蛋白和巨噬细胞特异性抗原在斑块内的表达。结果Compound 48/80对血清脂质水平无明显影响。Compound 48/80明显刺激肥大细胞脱颗粒(80.6%±17.8%比13.5%±4.1%,P<0.01),升高血清中类胰蛋白酶活性(0.57±0.13 u/L比0.36±0.10 u/L,P<0.05)。套环能加速颈总动脉斑块形成(未套环侧颈总动脉斑块面积均为0,套环侧颈总动脉均有斑块形成),Compound 48/80增大套环侧颈总动脉斑块最大横截面积(58 500±7 500μm2比8 600±2 800μm2,P<0.01),并使管腔最大狭窄程度加重(81%±15%比41%±12%,P<0.05)。Compound 48/80使颈总动脉斑块内α平滑肌肌动蛋白表达量增加(1 219±364 iu比522±137 iu,P<0.05)和巨噬细胞特异性抗原表达量增加(426±133 iu比169±38 iu,P<0.05)。结论套环能加速载脂蛋白E基因敲除小鼠颈总动脉斑块形成。Compound 48/80使套环侧颈总动脉斑块最大横截面积和颈总动脉最大狭窄程度增加,其机制可能与其刺激肥大细胞脱颗粒促使平滑肌细胞增殖和巨噬细胞聚集有关。     

血小板活化因子受体拮抗剂WEB2086抑制载脂蛋白E基因敲除鼠主动脉粥样硬化斑块内血管新生

目的　观察血小板活化因子受体拮抗剂WEB2 0 86对载脂蛋白E基因敲除鼠斑块内血管新生和斑块面积的影响。方法　8周龄雄性载脂蛋白E基因敲除鼠普通饲料喂养2 4周,然后随机分为对照组和WEB2 0 86组2组,每组18只。两组均普通饲料,对照组普通饮水,WEB2 0 86组则在普通饮水中加入血小板活化因子受体拮抗剂WEB2 0 86 (43mg/L)。继续喂养8周后,空腹18h采血,安乐死,取材。全自动生物化学分析仪检测血清脂质含量,CD31全层铺片检测斑块内毛细血管密度,含斑块的主动脉环置于matrigel中培养检测斑块新生毛细血管的能力,苏丹Ⅳ染色检测斑块面积。结果　WEB2 0 86对血清脂质无明显影响。WEB2 0 86明显抑制粥样斑块内毛细血管平均密度(34.6 %±10 .2 %比16 .1%±6 .7% ,P <0 .0 1)、明显抑制含斑块的主动脉环在matrigel中新生毛细血管(172 .3±4 0 .6比73.1±2 4 .9,P <0 .0 5 )、显著减小主动脉斑块面积(31.4 %±9.7%比17.5 %±6 .3% ,P <0 .0 1)。结论　WEB2 0 86对血脂无明显影响,但可抑制斑块内血管新生和减小斑块面积。     

血管内皮生长因子165对实验性兔动脉粥样硬化斑块形成的影响

目的观察血管内皮生长因子165对动脉粥样硬化斑块形成与发展的影响。方法利用高胆固醇饲料复制动脉粥样硬化兔模型。15只兔随机分为正常对照组、高胆固醇组和血管内皮生长因子组。42天时处死动物,截取胸主动脉进行计量组织学及免疫组织化学分析。结果正常对照组、高胆固醇组和血管内皮生长因子组的斑块面积(0%比1.81%±0.61%比24.12%±3.58%)、斑块周径(0比6.05%±1.62%比25.71%±1.97%)以及斑块最大厚度(0比0.06mm±0.002mm比0.16mm±0.007mm)均存在显著差异(P<0.05)。3组CD34阳性细胞数(cellsmm2)分别为0、12.35±2.02和61.15±7.55(P<0.05)。电镜显示新生血管与动脉粥样斑块相邻,新生血管腔内可见淋巴细胞。血管内皮生长因子组CD34阳性细胞数与斑块面积之间呈正相关(r=0.989,P<0.001)。结论血管内皮生长因子165能促进兔动脉粥样硬化斑块的形成与发展。     

罗格列酮增加apoE基因敲除小鼠主动脉粥样硬化斑块的稳定性研究

目的探讨罗格列酮对载脂蛋白E(apoE)基因敲除小鼠主动脉粥样硬化斑块稳定性的影响及其可能机制.方法32只6周龄apoE基因敲除小鼠随机分为罗格列酮组(18只)和对照组(14只),普通饮食14周后处死;切取小鼠主动脉根部制备石蜡切片,免疫组化法半定量测定两组粥样斑块内巨噬细胞、平滑肌细胞及炎性因子基质金属蛋白酶-9和肿瘤坏死因子-α的阳性率;Masson法半定量观察两组胶原纤维的灰度变化.结果罗格列酮组巨噬细胞的阳性率较对照组显著降低[分别为(16.1±2.5)%和(36.2±6.8)%,P＜0.05];平滑肌细胞的阳性率增加[分别为(38.5±7.2)%和(18.6±6.7)%,P＜0.05];胶原纤维的灰度显著降低[分别为(169.4±15.4)和(215.5±14.6),P＜0.05];基质金属蛋白酶-9分别为(16.0±4.8)%和(33.6±9.7)%,P＜0.05;肿瘤坏死因子-α的阳性率显著降低[分别为(15.9±2.9)%和(26.6±8.5)%,P＜0.05]. 结论罗格列酮可能通过下调局部炎性因子水平,改变斑块内的构成,稳定动脉粥样病变.     

半边莲生物碱抑制肾性高血压大鼠内皮素1 mRNA和蛋白表达

目的观察半边莲生物碱对肾性高血压大鼠内皮素1 mRNA表达、蛋白合成和释放的影响。方法采用两肾一夹高血压大鼠模型,随机分为高血压组、半边莲组、卡托普利组和假手术组。用原位杂交技术观察大鼠外周血白细胞内皮素1 mRNA的表达,应用免疫组织化学染色计数大鼠主动脉内皮细胞铺片内皮素阳性细胞率(反映内皮素1蛋白的合成),用放射免疫技术测定大鼠血浆内皮素的含量。结果与假手术组比较,高血压组外周血白细胞内皮素1mRNA表达增强(34.64%±8.39%比9.34%±4.47%,P<0.05),动脉内皮细胞合成内皮素增多(7.42%±0.24%比1.58%±0.24%,P<0.05),血浆内皮素水平显著升高(221±24ng/L比138±19ng/L,P<0.05)。应用半边莲生物碱8周后,与高血压组比较,内皮素1 mRNA表达(20.38%±11.31%比34.64%±8.39%,P<0.05)、内皮素合成(3.53%±0.21%比7.42%±0.24%,P<0.05)和血浆内皮素水平(191±21ng/L比221±24ng/L,P<0.05)均受到显著抑制。结论肾性高血压大鼠伴有内皮素表达增强,半边莲生物碱能抑制内皮素基因的转录、蛋白合成及翻译,对防治肾性高血压所致的血管病变具有一定作用。     

辛伐他汀消退老年载脂蛋白E基因敲除小鼠动脉粥样硬化斑块的观察

目的观察辛伐他汀对高脂喂养的老年载脂蛋白E(apoE)基因敲除小鼠动脉粥样硬化(AS)斑块的作用。方法选取高脂喂养的60周龄雄性aopE基因敲除小鼠30只,随机分为辛伐他汀高剂量组、辛伐他汀低剂量组和模型组,每组10只,分别每天予以辛伐他汀20mg/kg、辛伐他汀5mg/kg和等量生理盐水灌胃,连续灌胃8周后,取主动脉窦部做病理检测。冷冻切片光镜下观察AS斑块病理情况,免疫组织化学染色观察骨桥蛋白、α肌动蛋白的表达。vonkossa染色观察斑块钙化情况。结果与模型组比较,辛伐他汀高、低剂量组apoE基因敲除小鼠的血清TC和LDL-C水平明显降低(P<0.05),斑块面积占管腔面积比、斑块内钙化染色面积明显减少(P<0.05)。结论辛伐他汀可明显降低老年apoE基因敲除小鼠的血清TC和LDL-C水平,消退AS斑块的大小及其钙化。     

斑块内血管生成在兔动脉粥样硬化斑块发生发展中的作用

目的观察斑块内血管生成对兔动脉粥样硬化斑块形成与发展的影响。方法用高胆固醇饲料复制动脉粥样硬化兔模型。15只日本大耳白兔随机分为3组:A组,阴性对照组,仅给普通饲料喂养,B、C组给高胆固醇饲料喂养3周,A组及B组肌注白蛋白(2μg/kg)(0d),C组肌注血管内皮生长因子(VEGF1652μg/kg),继续以前饲养方式3周处死动物,截取胸主动脉进行计量组织学及免疫组织化学分析,测定不同组别不同时间点兔血清白细胞介素8(IL8)浓度和血脂浓度。结果(1)斑块面积(A组0,B组1.81%±0.61%,C组24.12%±3.58%)、斑块周径(A组0,B组6.05%±1.62%,C组25.71%±1.97%)及斑块的最大厚度(A组0,B组0.06mm±0.002,C组0.16mm±0.007mm),各组间比较有显著差异(P<0.05)。(2)新生血管的密度(CD34阳性细胞数细胞数/mm2(cells/mm2)A组0,B组12.35±2.02,C组61.15±7.55)各组之间比较有显著差异(P<0.05)。(3)电镜显示:新生血管与动脉粥样斑块相邻,新生血管腔内可见淋巴细胞。(4)血清IL8浓度(+21d时A组[0.05±0.006]pg/ml,B组[0.808±0.308]pg/ml,C组[15.72±4.31]pg/ml)各组间相比有显著差异。(5)此时血清胆固醇浓度B,C两组相比无显著差异。结论斑块内血管生成是动脉粥样斑块的重要病理特征,这个过程可能与炎性反应有关。     

甲珠对肝纤维化大鼠alpha-SMA和TGF-beta1表达的影响

目的: 研究甲珠对肝纤维化大鼠alpha-肌动蛋白(alpha-SMA)和转化生长因子-beta1(TGF-beta1)表达影响, 并探讨其抗纤维化机制. 方法: 采用400 mL/L CCL4 sc, 制备肝纤维化模型并以高[2.0 g/(kg·d)], 中[1.0 g/(kg·d)], 低剂量[0.5 g/(kg·d)]甲珠干预, 测定各组肝功能、血清TGF-beta1, 免疫组织化学法检测肝组织alpha-SMA、TGF-beta1表达, RT-PCR检测alpha-SMA、TGF-beta1mRNA 的表达. 结果: 甲珠各组较模型组肝功能明显改善, 丙氨酸转氨酶、天冬氨酸转氨酶显著降低, 总蛋白及白蛋白显著升高, 胆红素降低; 血清TGF-beta1显著降低; 高, 中, 低甲珠剂量组肝组织alpha-SMA及TGF-alpha1 mRNA表达, 染色面积比, 灰度值与模型组相比均有显著性意义(alpha-SMA: 9.21±1.12、12.63±2.42、14.23±1.57 vs 16.32±2.14, P<0.01、P<0.01、P<0.05; TGF-beta1 mRNA: 5.58±0.80、8.62±1.16、11.92±1.35 vs 14.57±1.59, P<0.01、P<0.01、P<0.01), (alpha-SMA染色面积比: 9.21%±1.29%、12.63%±1.44%、14.23%±1.41% vs 16.32%±1.75%, P<0.01、P<0.01、P<0.05; TGF-beta1染色面积比: 5.31%±0.70%、8.37%±1.09%、11.92%±1.42% vs 14.47%±1.48%, P<0.01、P<0.01、P<0.01), (alpha-SMA灰度值: 91.29±9.53、99.55±11.83、107.18±12.06 vs 116.44±12. 97, P<0.01、P<0.01、P<0.05; TGF-beta1灰度值: 89.96±9.64、106.92±13.90、110.50±12.91 vs 127.13±14.88, P<0.01、P<0.01、P<0.05). 结论: 甲珠对CCl4诱导的实验鼠肝纤维化有良好的抑治作用.     

中心型肥胖患者内皮舒张功能与血清瘦素相关

目的探讨中心型肥胖患者内皮依赖性舒张功能异常与血清瘦素的关系。方法按配对方法分为中心型肥胖组和对照组,用放射免疫法检测血清瘦素水平,用高分辨超声技术检测肱动脉内皮舒张功能。结果中心型肥胖组肱动脉内皮依赖性舒张功能较对照组明显减退(6.71%±3.60%比13.81%±3.71%,P<0.01),非内皮依赖性舒张功能差异无显著性(19.71%±6.63%比18.60%±6.35%,P>0.05)。中心型肥胖组血清瘦素水平明显高于对照组,男性肥胖组与男性对照组比较(8.63±3.73μg/L比3.05±1.56μg/L,P<0.01)、女性肥胖组与女性对照组比较(16.73±6.93μg/L比7.93±3.66μg/L,P<0.01)差异均有显著性。Pearson相关分析显示肱动脉内皮依赖性舒张功能与血清瘦素水平呈显著负相关。结论中心型肥胖者存在内皮依赖性舒张功能异常,它与血清瘦素水平的增高有关。     

RELMɑ/FIZZ1信号通路对载脂蛋白E基因敲除小鼠动脉粥样硬化斑块内血管新生的影响

目的 探讨类抵抗素分子ɑ或炎症区域分子1（RELMɑ/FIZZ1）对载脂蛋白E（ApoE）基因敲除小鼠动脉粥样硬化斑块稳定性及血管新生的影响及其信号通路。方法 8周龄C57BL/6J ApoE基因敲除鼠20只,喂食高脂饲料12周后随机分为模型组及RELMɑ/FIZZ1组,另选10只C57BL/6J野生型小鼠作为对照组;RELMɑ/FIZZ1组于尾部血管注射重组RELMɑ/FIZZ1干预2周后结束实验。取小鼠主动脉制备石蜡包埋切片,进行HE染色,利用图像软件定量测量斑块面积、血管横截面积及校正斑块面积,采用免疫组织化学染色测定主动脉血管壁RELMɑ/FIZZ1及CD34阳性反应强度。提取主动脉RNA,采用全基因表达谱筛选出显著表达差异的基因和发生变化的细胞通路。结果 与对照组相比,模型组动脉粥样硬化明显,斑块面积增加,粥样硬化斑块内RELMɑ/FIZZ1表达明显。RELMɑ/FIZZ1刺激后RELMɑ/FIZZ1及CD34阳性反应强度增强,校正斑块面积比模型组显著性增加（31.58%±6.65%比24.16%±3.59%,P<0.01）,明显刺激血管新生（P<0.05）。相对于对照组,RELMɑ/FIZZ1组有显著性上调基因391个,下调基因465个;活性显著性上调信号通路12条,活性显著性下调信号通路10条,共计22条。结论 RELMɑ/FIZZ1刺激血管新生,造成粥样斑块不稳定,其机制与Atg9a、Gng8等基因显著性表达及细胞肌动蛋白骨架调节通路、缝隙连接信号通路的激活密切相关。     

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.