酰基化Ghrelin在体外拮抗巨噬细胞介导的炎症反应对胰岛β细胞功能影响的研究

目的 采用Transwell小室构建的小鼠胰岛细胞株MIN6和巨噬细胞株RAW264.7共培养系统,探讨酰基化Ghrelin能否保护胰岛β细胞免遭巨噬细胞介导的炎症损伤. 方法 实验分为单独RAW264.7巨噬细胞组、单独MIN6胰岛细胞组、共培养组和Ghrelin干预组.RT-PCR和Western blot 检测巨噬细胞上Toll样受体4(TLR4)和脂肪酸结合蛋白(A-FABP)的表达;ELISA检测细胞上清液IL-1β、TNF-α的浓度,以及葡萄糖刺激后MIN6细胞上清液的胰岛素浓度. 结果 (1)共培养系统中,巨噬细胞TLR4[mRNA及蛋白水平分别为(1.35±0.13),(0.93±0.03)],A-FABP[(1.99±0.10)vs(0.91±o.01)]的表达],细胞上清液IL-1β[(10.47±1.11) pg/ml]、TNF-α[(917.54±9.09) pg/ml]蛋白水平较单独RAW264.7巨噬细胞组高(P＜0.05);(2)高糖刺激下,共培养系统中胰岛β细胞的胰岛素分泌水平较单独MIN6胰岛细胞组低(P＜0.05);(3)与共培养组比较,酰基化Ghrelin干预后共培养,巨噬细胞TLR4表达水平和TNF-α均降低(P＜0.05),且呈剂量依赖性,但胰岛β细胞的胰岛素分泌水平与共培养组比较,差异无统计学意义(P＞0.05). 结论 巨噬细胞与胰岛β细胞共培养后,炎症通路活化,炎症因子释放,胰岛β细胞的胰岛素分泌功能受损;酰基化Ghrelin可剂量依赖性削弱巨噬细胞和胰岛β细胞共培养后炎症通路的活化和炎症因子的释放,但不能完全阻止胰岛β细胞胰岛素分泌功能的降低.     

低密度脂蛋白免疫复合物对巨噬细胞细胞因子分泌的影响

目的探讨低密度脂蛋白免疫复合物(LDL-IC)对巨噬细胞细胞因子分泌的影响.方法设立终浓度均为150 μg/ml的LDL组、LDL-IC组、Igg-IC组及阴性对照组与细胞孵育相同时间,以ELISA法检测比较培养上清TNF-α和IL-1β水平.并观察不同LDL-IC浓度(0,50,100,150,200,250μg/ml)作用相同时间以及终浓度为150μg/ml的LDL-IC与细胞作用不同的时间(0,3,6,12,24,36 h),培养上清TNF-α和IL-1β水平.结果LDL-IC组培养上清TNF-α、IL-1β水平均明显高于LDL、IgG-IC及阴性对照组.TNF-α、IL-1β水平随着LDL-IC作用浓度的增加和作用时间的延长而增高.结论LDL-IC可以显著增加细胞TNF-α和IL-1β的分泌,具有剂量和时间依赖效应.提示LDL-IC能通过增加细胞因子分泌而在动脉粥样硬化中起重要作用.     

脂多糖对大鼠肺泡巨噬细胞分泌一氧化氮和氧化应激的影响

目的探讨细菌内毒素脂多糖（LPS）对SD大鼠肺泡巨噬细胞产生一氧化氮（NO）和氧化应激的影响。方法采用支气管肺泡灌洗和细胞差速贴壁的方法分离大鼠肺泡巨噬细胞（AM）,分别测定AM培养上清液NO含量、一氧化氮合酶（NOS）活性、丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性。结果在5,10,20,50mg/LLPS分别干预下,大鼠AM培养上清液NO含量、NOS活性和MDA含量均显著升高,SOD活性显著降低,并且具有浓度依赖性。结论LPS促进大鼠AM分泌NO,并诱导AM脂质过氧化损伤,这可能是内毒素诱发肺部炎症反应不易控制和急性肺损伤的机制之一。     

血浆巨噬细胞刺激蛋白表达在AECOPD的临床研究

目的研究患者血浆巨噬细胞刺激蛋白(macrophage stimulating protein,MSP)表达在AECOPD的临床研究。方法随机选择我院收治的60例急性加重期COPD(AECOPD)患者,60例稳定期COPD患者,并选择年龄、性别配对的60例正常体检人员设为对照组。检测三组受试者血浆MSP水平以及白介素6(in-terleukin-6,IL-6)、IL-8等炎性因子水平,并进行MSP水平与血清炎性因子水平及肺动脉压的相关性分析。结果 AECOPD组血浆MSP水平显著高于稳定期COPD组与对照组,稳定期COPD组显著高于对照组,(P 0. 05); AECOPD组血清IL-6、IL-8、肿瘤坏死因子α(Tumor necrosis factorα,TNF-α)水平显著高于稳定期COPD组与对照组,稳定期COPD组显著高于对照组,(P 0. 05);血浆MSP表达水平与血清IL-6、IL-8、TNF-α水平、平均肺动脉压(mm Hg)呈显著正相关(P 0. 05)。结论 MSP与COPD有关,与急性加重有关,可能与MPS通过调控巨噬细胞参与COPD气道炎症反应有关,其表达水平变化有助于COPD诊断与治疗指导。     

半夏泻心汤抑制巨噬细胞分泌促炎因子抗胃炎机制研究

[目的]探讨中国经典方半夏泻心汤调节固有免疫细胞巨噬细胞分泌促炎性细胞因子抗胃炎的机制。[方法]采用含药血清作用原代培养的巨噬细胞,用荧光定量PCR的方法检测巨噬细胞内IL-8、IL-12、IL-18、TNF-α表达变化情况结果作用3h时,5%含药血清对巨噬细胞分泌IL-8、IL-12、IL-18无影响,能抑制巨噬细胞分泌TNF-α(P0.05);10%含药血清能促进巨噬细胞分泌IL-8、IL-18、TNF-α(P0.01),对巨噬细胞分泌IL-12仍无影响。作用6h时,5%含药血清能明显抑制巨噬细胞分泌IL-8(P0.01)、同时促进IL-18、TNF-α分泌(P0.05),对IL-12分泌无影响;10%含药血清能明显抑制巨噬细胞分泌IL-8、IL-18、TNF-α(P0.01),对IL-12分泌仍无影响。对于IL-8的影响,只有在作用6h时呈现剂量—效应关系。对于IL-18的影响,有明显的时间-剂量依赖性,含药血清作用时间越长,作用浓度越高,对抑制巨噬细胞分泌细胞因子作用越明显。对于TNF-α的影响,只有在作用6h时呈现剂量—效应关系,无时间—效应关系。[结论]半夏泻心汤可以有效地治疗因幽门螺杆菌感染导致的胃炎,其作用机制为通过调节固有免疫细胞巨噬细胞的活性,抑制巨噬细胞分泌IL-8、IL-18、TNF-α等炎性因子,减轻胃黏膜上皮细胞的炎性损伤。     

阿魏酸钠对结肠炎大鼠结肠巨噬细胞功能的影响

目的探讨阿魏酸钠在整体水平对结肠炎大鼠结肠巨噬细胞功能的影响及其机制。方法建立大鼠免疫性结肠炎模型。阿魏酸钠（SF）灌肠用药21d后检测结肠组织MDA、NO、PGE2含量，SOD、IL-1，TNF-α、MPO活性及NF-κB p65表达水平。结果阿魏酸钠（200，400，800mg／kg）灌肠后呈剂量依赖型降低模型组大鼠的MDA、NO、PGE2含量，IL-1、TNF—α、MPO活性及NF-κB p65表达水平，同时升高SOD活性。结论SF可减弱结肠炎大鼠结肠活化巨噬细胞的生物活性，缓解结肠炎症反应，机制可能与抑制NF-κB表达有关。     

双氢青蒿素治疗对卡氏肺孢子虫肺炎大鼠肺泡巨噬细胞上清液TNF—α和NO水平的影响

目的 研究双氢青蒿素治疗对卡氏肺孢子虫肺炎大鼠泡巨噬细胞上清液TNF－α和NO水平的。方法 以醋酸可的松皮下注射Wistar大鼠建立卡氏肺孢子虫肺炎动物模型。用60mg/kg双氢青蒿素治疗实验大鼠后杀鼠取,用胶原酶消化法分离肺泡巨噬细胞,用LPS刺激培养72h,同时庙有感染组和正常对照。用TNF－α和NO试剂盒分别检测培养上清液TNF－α和NO活性。结果 感染组和治疗组TNF－α和NO水平均高于正常对照,治疗组TNF－α和NO水平则低于感染组。结论 卡氏肺孢子虫感染引起大鼠肺泡巨噬细胞分泌高水平的TNF－α和NO,胆双氢青蒿素治疗后PCR大鼠肺泡巨噬细胞产生TNF－α和NO水平降低。     

乌司他丁对脂多糖诱导的巨噬细胞炎症反应的影响

目的探讨乌司他丁对LPS诱导的RAW264.7细胞炎症反应的影响。方法采用1.0μg/mL的LPS激活RAW264.7细胞株,与不同浓度组乌司他丁(100～10 000 U/mL)共同孵育,采用ELISA法测定TNF-α和IL-1β的表达;实时荧光定量逆转录聚合酶链反应(RT-PCR)技术分析TNF-αmRNA和IL-1βmRNA表达。结果高浓度乌司他丁(1 000～10 000 U/mL)可抑制LPS诱导的RAW264.7细胞TNF-α和IL-1β表达(P均<0.05),下调LPS诱导的RAW264.7细胞TNF-αmRNA和IL-1βmRNA含量(P均<0.05);低浓度乌司他丁(100 U/mL)对LPS诱导的RAW264.7细胞TNF-α、IL-1β、TNF-αmRNA、IL-1βmRNA表达与LPS单独处理组比较,无显著差异。结论乌司他丁可抑制LPS诱导的巨噬细胞炎症反应;该抑制作用呈浓度依赖性。     

普萘洛尔对BALB/C小鼠腹膜巨噬细胞分泌细胞因子及吞噬功能的影响

目的:探讨普萘洛尔对脂多糖诱导BALB/C小鼠腹膜巨噬细胞分泌肿瘤坏死因子(TNF)-α和白细胞介素(IL)-10以及吞噬功能的影响。方法:将腹膜巨噬细胞分为8组:空白组,脂多糖组,普萘洛尔(0.1μmol/L)组,普萘洛尔(1.0μmol/L)组,普萘洛尔(10.0μmol/L)组;普萘洛尔(0.1μmol/L)加脂多糖组,普萘洛尔(1.0μmol/L)加脂多糖组,普萘洛尔(10.0μmol/L)加脂多糖组。在培养6h和24h后用酶联免疫法测定各培养瓶上清液中TNF-α和IL-10的浓度,用中性红吞噬试验检测巨噬细胞吞噬功能。结果:空白组与单用普萘洛尔时,巨噬细胞均无细胞因子分泌(无数据显示),吞噬功能无明显影响。单用脂多糖组巨噬细胞释放TNF-α和IL-10显著增加,吞噬能力明显增强。而普萘洛尔加脂多糖组巨噬细胞继续分泌TNF-α,IL-10分泌减少,吞噬功能显著降低,该作用随普萘洛尔浓度增加而增强。结论:普萘洛尔可调节腹膜巨噬细胞炎症递质的分泌,并能影响腹膜巨噬细胞的吞噬能力。     

双氢青蒿素对患卡氏肺孢子虫肺炎大鼠血清和肺泡巨噬细胞上清液IL-1水平的影响

为研究双氢青蒿素对患卡氏肺孢子虫肺炎（Pneumocystis carinii pneumonia,PCP）大鼠血清和肺泡巨噬细胞培养上清液IL－1水平的影响，以醋酸可的松皮下注射Wistar大鼠建立卡氏肺孢子虫肺炎动物模型，用60mg/kg双氢青蒿素治疗实验大鼠，杀鼠取肺，用胶原酶消化法分离大鼠肺泡巨噬细胞，用LPS刺激培养72h，同时设有感染对照组和正常对照，用IL－1β试剂盒分别检测血清和培养上清液IL－1β的水平，结果显示感染组和治疗组大鼠IL－1β水平显著高于正常对照，治疗组大鼠IL－1β水平则低于感染组，说明卡氏肺孢子虫感染可能引起大鼠肺泡巨噬发泌高水平IL－1，经双氢青蒿素治疗PCP后大鼠肺泡巨噬细胞产生IL－1水平降低。     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

Angiographic diagnosis of the degree of serosal invasion of carcinoma of the colon

Angiography using Prostaglandin El^® was performed on 38 patients with carcinoma of the colon in order to diagnose the degree of serosal cancer invasion. The findings at angiography were classified into four groups:1) AG-S3, abnormal change (irregularity and/or encasement) up to marginal vessels; 2) AG-S2, abnormality up to vasa recta; 3) AG-S1, abnormality of penetrating branches of vasa recta within the wall of the colon; and 4) AG-S0, no distinct findings of abovementioned vessels. These angiographic findings were compared with both macroscopic and microscopic serosal cancer invasion. Angiographic diagnosis is in accord with the macroscopic findings in 84.2 percent of cases. Angiographic diagnosis is in accord with the microscopic findings in 32.4 percent of cases. Macroscopic findings confirm the angiographic diagnosis precisely but the conflict with microscopic findings should not be overlooked. This may be the result of inflammatory change, adhesion, and fibrosis around carcinoma of the colon.