贝氏柯克斯体重组蛋白对树突状细胞活化作用分析

目的 分析贝氏柯克斯体(Cb)的Com1和热休克蛋白B(HspB)对体外诱导培养的人源树突状细胞(DC)表面分子和细胞因子表达的影响.方法 分别以20 μg/mL的重组蛋白Com1和HspB、6μg/mL的大肠埃希菌LPS为终浓度刺激培养5d的DC,24 h后流式细胞仪检测DC表面成熟标志分子CD83和T淋巴细胞CD58、CD54、CD40、CD80、CD86以及细胞因子IL-10和IL-12的表达水平.多重比较应用SNK检验.结果 Com1刺激能有效促进体外诱导培养的DC成熟,DC细胞表面成熟标志分子CD83和T淋巴细胞活化辅助分子CD54、CD58、CD80、CD86和CD40等处于高表达水平,表达水平均＞80％,其中CD83表达水平与大肠埃希菌LPS刺激DC相似,CD54和CD86的表达量略高于大肠埃希菌LPS刺激DC,其他分子的表达量显著高于大肠埃希菌LPS刺激DC(均P＜0.05).Com1刺激后,细胞内IL-12水平从无增至9％左右.HspB刺激不能促进DC表型成熟,HspB刺激的DC胞内IL-10水平达6％左右.Com1和HspB的先后刺激,使IL-12水平几乎为0,IL-10水平降至2％以下.结论 Com1为DC有效的促成熟抗原,被Com1激活的DC具备启动T淋巴细胞免疫的功能条件.     

乙型肝炎病毒对树突状细胞表型成熟和功能活化的影响

目的：研究乙型肝炎病毒（HBV）病毒粒子和抗原成分对树突状细胞（DC）成熟和功能活化的影响。方法：分离培养小鼠骨髓来源的DC细胞（BM-DC）,在培养过程中加入含有HBV病毒粒子和抗原成分的培养基,流式细胞技术分析DC细胞表面分子的表达水平,病毒保护实验和细胞因子生物分析方法分别检测DC细胞培养上清中的干扰素（IFN）和肿瘤坏死因子（TNF）的表达水平。结果：培养基中加入HBV病毒粒子和抗原成分显著抑制DC细胞表面共刺激分子（CD40、CD80、CD86）和成熟标志分子（MHC—Ⅱ）的表达,降低DC细胞培养上清中IFN和TNF的水平。结论：HBV可以直接抑制DC的表型成熟和功能活化,有可能在HBV感染的慢性化机制中起重要作用。     

糖尿病患者外周血树突细胞功能活性检查及抑制NF-κB活性的意义

目的 检测糖尿病(DM)患者外周血单个核细胞(PBMNCs)培养树突细胞(DC)的功能表达及其临床意义, 探讨DC成熟过程中核因子 (NF)-κB基因的作用.方法 DM患者及健康成人PBMC体外经GM-CSF和IL-4诱导培养DC,在DC培养中加入NF-κB特异性低脱氧核苷酸(ODN)阻断NF-κB活性,检测细胞表面分子表达以及混合淋巴细胞反应(MLR)检测其功能变化.结果 DM患者外周血培养获成熟DC数量较多,DC表面标志较正常对照表达升高,具有较强的激发MLR的能力,DC成熟高于正常人.但NF-κB ODN可以抑制DC表面标志的表达,阻碍DC发育成熟,降低激发MLR的能力,并不可被脂多糖(LPS) 所逆转.结论 DM患者DC免疫功能高于正常对照DC功能状态,NF-κB是DC发育成熟过程中关键性调控基因.应用NF-κB ODN可抑制DC成熟,诱导生成耐受原性未成熟DC.     

氧化苦参碱对小鼠树突状细胞成熟和功能的影响

目的:研究氧化苦参碱(OXY)对小鼠树突状细胞(DC)成熟、表型及功能的影响。方法:流式细胞术检测DC表面分子CD40的表达;混合淋巴细胞反应(MLR)检测DC对T淋巴细胞的刺激能力;ELISA法检测MLR上清中细胞因子IFN-γ的分泌。结果:第0天OXY处理组较对照组DC表面分子CD40的表达明显升高(P<0.01),刺激T细胞能力增强,分泌细胞因子IFN-γ明显升高(P<0.05),对LPS诱导的DC成熟,与DC LPS组对照显著升高(P<0.05)。结果:OXY对DC的成熟和功能有一定的促进作用。     

负载HBcAg对慢性HBV感染患者树突状细胞的活化作用及功能影响

目的:探讨免疫耐受期慢性乙型肝炎(CHB)患者外周血树突状细胞(DC)负载HBcAg后细胞表型及免疫功能的改变。方法:从CHB免疫耐受期患者外周血分离培养DC,在DC成熟前,加入重组HBcAg表位肽,诱导HBV特异性DC分化成熟,流式细胞仪检测DC表面共刺激分子CD1a、CD80、CD83的表达水平,应用淋巴细胞增殖试验评估DC功能。结果:负载不同剂量的HBcAg后,DC细胞活化增强,DC细胞共刺激分子标志物CD1a、CD80以及CD83表达率均明显上升,且随着抗原负载剂量的增加,表达率进一步增加,各组之间差异具有统计学意义(均P<0.001);未负载HBcAg的DC细胞激活的淋巴细胞反应较弱,负载HBcAg的DC细胞刺激同种异体健康成人T淋巴细胞增殖能力增强,且随着负载抗原剂量的加大,T淋巴细胞的增殖能力进一步提高,与未负载抗原的对照组相比,差异具有统计学意义(F=428.14,P=0.000)。结论:体外负载HBcAg刺激CHB患者DC细胞可增强其有效抗原提呈能力,促进T淋巴细胞增殖。     

雷帕霉素对人树突状细胞功能的影响

目的　研究雷帕霉素 (RPM)对人树突状细胞 (DC)功能的影响。方法　分离外周血单个核细胞 (PBMC) ,在含粒细胞巨噬细胞集落刺激因子 (GM CSF)、白介素 4 (IL 4 )以及有或无RPM的培养条件下制备DC。用流式细胞仪检测DC表面共刺激分子CD86 (B7 2 )的表达 ;混合淋巴细胞反应(MLR)检测DC对同种异体T淋巴细胞的刺激能力 ;ELISA法测定MLR上清液中的细胞因子。结果　与对照组比较 ,经RPM处理的DC表面CD86的表达明显降低 ;对T淋巴细胞刺激的能力下降 ;MLR中致炎细胞因子 (IL 1β ,IL 6 ,TNF a)浓度降低。RPM的这些作用呈浓度依赖性。 结论　雷帕霉素对人树突状细胞功能有明显的抑制作用 ,可能是其防止支架术后再狭窄的机制之一     

慢性粒细胞白血病树突状细胞激活的特异性抗白血病免疫反应的体外研究

树突状细胞 (DC)是目前发现的抗原递呈功能最强的抗原递呈细胞 (APC)。本实验从慢性粒细胞白血病 (CML)患者外周血单个核细胞 (PBMNC)诱导生成既具有特异抗原 ,又表达共刺激分子的DC ,探索其在诱导特异性抗CML免疫反应中的作用。一、材料和方法1 样本来源 :初诊的CML患者 11例 ,经逆转录 聚合酶链反应 (RT PCR) ,检测bcr/abl融合基因均为阳性。2 DC的培养与鉴定 :CML患者PBMNC以 10 6/ml浓度 ,在含有重组人粒细胞巨噬细胞集落生长因子 (rhGM CSF) 10 0 μg/L、白细胞介素 4 (IL 4 ) 5 0 0U/ml、肿瘤坏死因子α(TNFα)…     

胆汁酸膜受体TGR5在小鼠骨髓来源的树突状细胞中的表达及其相关功能

目的 通过TGR5天然激动剂石胆酸(LCA)刺激小鼠树突状细胞(DC),探讨TGR5受体在DC成熟活化及吞噬功能方面的作用.方法 体外诱导小鼠骨髓细胞形成DC,LCA 30 μmol/L预处理2h后,用CFSE标记的胸腺凋亡细胞测试其吞噬功能;加入脂多糖(LPS)1 μg/mL诱导其成熟,6h后提取细胞总RNA,PCR法检测IL-12、TNF-α和IL-10等细胞因子;48 h后以流式细胞术检测细胞表面分子MHCII、CD80、CD86表达水平.组间比较采用t检验.结果 (1)LCA可以上调DC表达TGR5,并增强DC吞噬凋亡细胞功能(P＜0.05);(2)LCA可以降低LPS引起的DC表面MHCⅡ、CD80和CD86表达水平(均P＜0.05),提示LCA可抑制DC成熟;(3)LCA可抑制LPS诱导的DC促炎性细胞因子IL-12、IL-6、TNF-α mRNA表达,而IL-10表达水平升高(均P＜0.05).结论 TGR5受体激动剂可抑制DC成熟并增强DC吞噬功能,发挥免疫调节作用.     

ADMA对单核细胞来源的树突状细胞成熟和免疫的影响

目的:通过观察非对称性二甲基精氨酸(ADMA)对树突状细胞(dendritic cells,DCs)成熟及免疫的影响来探讨AS形成的可能机制。方法:贴壁法分离人外周血单核细胞,在含重组人粒-巨噬细胞集落刺激因子(rhGM-CSF 20ng/ml)和重组人白细胞介素-4(rhIL-4,10ng/ml)的完全培养基中培养,五天后收集imDC,用1、8、16umol/L的ADMA干预未成熟DC 24h。用流式细胞术检测DC细胞表面分子的表达、吞噬能力及DC的凋亡,用混合淋巴细胞反应检测成熟DC刺激T淋巴细胞增殖的能力,用ELISA检测DC细胞因子的分泌。结果:生理浓度ADMA并不刺激DC成熟及分化;但病理浓度ADMA抑制DC成熟;抑制DC诱导的T淋巴细胞增殖;诱导DC凋亡:抑制DC分泌IL-12细胞因子、TNF-α及IL-10细胞因子。结论:生理浓度ADMA并不刺激DC成熟及分化;但病理浓度AD-MA抑制DC成熟和免疫。     

贝氏柯克斯体新桥株刺激人树突状细胞的表型分析

目的树突状细胞(dendritic cell,DC)具有成熟与未成熟两种形态,前者是早期免疫应答强有力的抗原呈递细胞,后者能诱导免疫耐受。分析贝氏柯克斯体(Coxiella burnetii,Cb)、Cb脂多糖(LPS)、去除LPS的Cb对DC的激活作用,探讨Cb逃逸宿主免疫清除的机理。方法从健康成年人的外周血中分离单核细胞,加入GM-CSF和IL-4体外培养单核细胞成未成熟DC;分别用灭活Cb、Cb LPS、去LPS Cb刺激体外培养未成熟DC,大肠杆菌LPS作为阳性对照刺激因子;刺激后24h,用流式细胞仪(fluorescence activated cell sorter,FACS)检测DC表面分子:成熟标记分子CD83与共刺激分子CD40、CD58、CD80、CD86。结果去LPS Cb刺激DC表面分子表达水平与大肠杆菌LPS刺激最相近,显著强于未刺激DC,而灭活Cb、Cb LPS刺激DC表面分子CD83、CD40、CD80、CD86的表达在一低水平。结论LPS遮盖了Cb激活DC所需的表面分子,除去LPS后暴露的分子与DC相互作用,对DC的成熟刺激作用显著高于灭活Cb、Cb LPS的刺激作用。LPS的存在有利于贝氏柯克斯体逃逸宿主的免疫清除。     

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.     

Study of constancy of hydrogen-consuming flora of human colon

The constancy of the hydrogen consuming flora of the human colon was studied in 15 healthy subjects via two measurements obtained 18 to 36 months apart. Hydrogen disappearance rate and the major products of H₂-consuming bacteria, methane and sulfide, were measured during incubation of fecal homogenates with excess hydrogen and sulfate. In 11/15, the hydrogen consumption rate and the predominant hydrogen-consuming pathway (methanogenesis, sulfate reduction, or neither) remained constant. However, major shifts in these pathways were observed in four subjects, with two losing and two gaining the ability to produce methane. Methanogenesis was associated with the highest hydrogen consumption rate. This study demonstrates that clinically unrecognizable, major alterations of the colonic flora occur in healthy subjects. Understanding of the factors responsible for these alterations might allow for therapeutic manipulation of the colonic flora.Supported in part by the Department of Veterans Affairs and NIDDKD RO1 DK 13309-25.