系统性红斑狼疮的基因表达谱及其免疫调控通路的研究

目的 通过基因表达谱勾画系统性红斑狼疮（SLE）发病机制中可能的免疫调控通路。方法 用寡核苷酸基因芯片研究了10例SLE（包括2例SLE同胞对）和10份正常对照的外周血白细胞基因表达谱。进行了基因表达谱和临床免疫表型的聚类分析和比较，筛选统计学显著性差异表达的SLE相关基因，并在转录和蛋白表达水平验证芯片表达谱结果。结果 聚类分析揭示SLE临床免疫表型与基因表达谱特征存在关联。获得25个显著性表达的SLE相关基因，其中大部分未经报道。转录水平直接验证了其中C／EBPD，LY6E，OAS2三个基因的表达，较大样本独立验证了C／EBPD，LY6E在SLE中的表达上调；蛋白质水平验证了C／EBPD的高表达。结论 SLE基因表达谱可能是其临床免疫表型的分子基础，探讨了通过基因表达谱进行SLE分型乃至鉴别诊断的可能性。通过显著性差异表达的SLE相关基因，推测干扰素及其免疫调控通路在SLE发病中可能扮演重要角色。     

慢性阻塞性肺疾病患者T淋巴细胞氧运输相关基因的初步研究

目的利用基因芯片研究慢性阻塞性肺疾病（COPD）患者T淋巴细胞氧运输相关基因。方法采集COPD患者和健康人的外周血作为实验血样和对照血样。用Ficoll技术分离外周血淋巴细胞,用分选式流式细胞仪分选并纯化T淋巴细胞,并采用一步法提取总RNA,逆转录合成双链cDNA后,体外转录合成生物素标记的cRNA,片断化后,采用人类全基因表达谱芯片进行芯片杂交,扫描后筛选出差异表达基因。结果初步筛选出COPD／健康人外周血T淋巴细胞氧运输相关高表达差异基因3条。结论基因芯片技术能有效地研究基因的表达谱,筛查出COPD患者T淋巴细胞氧运输相关差异表达基因。     

基因芯片在肺癌发生相关基因筛查中的应用

目的 研究人肺癌发生相关基因表达谱，探讨肺癌发生的分子机制。方法 选取1280个与肿瘤相关的基因克隆。制备成肿瘤相关cDNA芯片。提取人肺癌组织以及相应正常组织RNA，反转录后标记为cDNA探针，与cDNA芯片杂交，经扫描及Quantarray3．0软件分析后比较两种组织中的差异表达基因。结果 共筛查出差异表达基因52条。其中表达上调基因35条，下调基因17条；按照基因功能可分为运输载体，代谢相关基因、细胞信号转导分子、细胞骨架、转录调控因子以及未知功能基因。结论 基因芯片可用于肺癌相关基因表达谱的筛查，为明确肺癌发生机制提供重要参考。     

应用基因芯片检测系统性红斑狼疮患者外周血B淋巴细胞

目的 应用寡聚核苷酸基因表达芯片研究系统性红斑狼疮(SLE)患者与健康志愿者外周血B淋巴细胞基因表达谱的变化.方法 应用包含21522条70 mer长度Oligo DNA的寡聚核苷酸芯片(22K Human Genome Array)分别检测6例SLE患者和6名健康志愿者外周血B淋巴细胞基因表达谱,应用聚类分析法分析差异表达的基因.结果 SLE患者外周血B淋巴细胞和健康志愿者B淋巴细胞之间基因表达谱有显著差异,芯片中聚类表达显著上调的基因15个,主要包括Ⅰ型干扰素(IFN)通路中的相关分子;聚类表达显著下调的基因22个,主要包括神经内分泌相关的精氨酸甲基化转移酶家族等与雌激素代谢相关的基因.进一步对这些差异表达基因进行了初步的功能分类分析,分析结果表明这些基因主要与B淋巴细胞的JAK-STAT信号通路和激素代谢相关.结论 SLE患者外周血B淋巴细胞基因表达谱存在显著差异,芯片显示氨基酸代谢、激素代谢、细胞增殖和凋亡、炎症反应等相关生物学功能的改变可能参与SLE的发病.     

应用基因芯片技术筛选肝细胞癌相关基因

目的:探讨基因表达谱芯片技术在筛查肝细胞癌(HCC)相关基因群表达中的作用。方法:TRIzol法抽提2例HCC及正常肝脏组织总RNA,分离纯化两种组织的mRNA;逆转录合成掺入生物素标记的cDNA合成探针,与基因芯片(涵盖了18400个转录本,代表了14 500个明晰的基因)杂交,扫描芯片荧光信号图像,计算机分析,比较2种组织基因表达谱差异。结果:2例HCC组织与正常肝脏组织相比,有2756条基因(19.01%)共同表达差异,其中共同上调基因1772条和共同下调基因984条,对2756条共同差异表达基因作了初步功能分类,这些基因与HCC的发病机制存在相关性。结论:基因表达谱芯片技术可以筛选出HCC表达异常的相关基因群,有助于认识肝癌的发病机制。     

应用基因芯片技术筛选肝硬化相关基因

目的:探讨基因表达谱芯片技术在筛查肝硬化相关基因群表达中的作用.方法:按TRIzol法抽提肝硬化及正常肝脏组织总RNA,分离纯化两种组织的mRNA.经逆转录合成掺入生物素标记的cDNA合成探针,与基因芯片(涵盖18 400个转录本,代表14 500个明晰的基因)杂交,扫描芯片荧光信号图像,计算机分析,比较二种组织基因表达谱差异.结果:2例肝硬化组织与正常肝脏组织相比,有1424条基因(9.82%)共同表达差异,其中共同上调基因980条和共同下调基因444条.对1424条共同差异表达基因作了初步功能分类,这些基因与肝硬化的发病机制存在相关性.结论:基因表达谱芯片技术可以筛选出肝硬化表达异常的相关基因群,对其进一步研究有助于认识肝硬化的发病机制.     

基因芯片技术对哮喘病人相关基因的筛选和分析

目的 利用基因芯片技术寻找哮喘病患者与正常人外周血单核细胞之间差异表达基因,拟为哮喘的早期诊断及预防提供分子标记.方法 用淋巴细胞分离液分别提取16例哮喘病患者与16例正常人外周血单核细胞,用QIAGEN Rneasy Kit提取纯化样本总RNA,并合成用荧光标记的cRNA,分别与含有41 000条基因序列的全基因芯片杂交,以基因表达倍数值≥2.0和基因表达倍数值≤-2.0为阈值来确定差异表达基因,然后用Genespring软件利用生物信息学方法对差异表达基因进行功能分类分析.结果 按P＜0.05差异显著性标准,从34 183条表达基因谱中,筛选出哮喘患者与正常对照差异表达2倍以上的基因有4177条,差异表达2倍以上已知与哮喘相关的基因有19条.经代谢途径分析发现这些差异基因主要涉及到炎症反应、免疫反应、防御反应、创伤反应、外部刺激反应等8大功能分类.结论 哮喘的发生涉及众多基因表达的改变,芯片技术可以有效地筛选出哮喘患者与正常对照的差异表达基因,对进一步探索哮喘的发病机制、有效的干预或逆转哮喘具有重要意义.     

重症肌无力患者胸腺自身免疫相关基因表达的研究

目的探讨MG患者和正常对照者的胸腺组织炎症反应与自身免疫性疾病相关基因的表达情况。方法取MG患者（MG组）和正常对照者（对照组）的胸腺组织标本，提取总RNA，反转录合成cDNA后，体外转录合成生物素标记的cRNA与基因芯片进行杂交，通过化学发光法检测基因表达，比较两组胸腺组织基因表达的差异。结果MG组与对照组比较，有61条基因存在显著性差异，其中表达上调33条，下调28条，这些基因包括炎症相关趋化因子基因、细胞因子及其受体基因、与细胞因子产生与代谢有关的基因、细胞因子及其受体相互作用相关的基因、炎症反应相关基因以及体液免疫相关基因等。结论芯片筛选MG患者胸腺自身免疫相关基因表达与正常对照组存在显著差异。     

应用TaqMan实时荧光定量PCR技术研究系统性红斑狼疮疾病相关基因的表达

目的验证新发现的4个系统性红斑狼疮(SLE)相关基因与SLE发病的相关性,为SLE发病的分子机制研究提供资料。方法前期基因芯片工作中筛选得到4个在55例SLE患者及正常对照之间表达差异显著的基因,染色体定位发现其位于SLE连锁的染色体区段。应用TaqMan实时荧光定量聚合酶链反应(PCR)技术检测该4种基因在SLE患者和正常对照外周血单核细胞及T、B淋巴细胞的表达。结果SLE患者外周血单核细胞中,TRIM基因ΔCt值显著低于正常人对照(P＜0．05),基因表达水平为正常对照的0．61倍；IL-4R基因与EGR1基因的ΔCt值显著高于正常人对照(P＜0．05),基因表达水平分别为正常对照的2．62、2．03倍；KLRC1基因的ΔCt值显著高于正常人对照(P＜0．01),基因表达水平为正常对照的5．33倍。在SLE患者外周血T细胞,TRIM基因ΔCt值显著低于正常人对照(P＜0．01),基因表达水平为正常对照的0．56倍；IL-4R基因与EGR1基因的ΔCt值显著高于正常人对照(P＜0．05),基因表达水平分别为正常对照的2．55、1．74倍；KLRC1基因的ΔCt值显著高于正常人对照(P＜0．01),基因表达水平为正常对照的5．19倍。在SLE患者外周血B淋巴细胞,由于B细胞淋巴所占淋巴细胞比率过低,基因检测数据不稳定,故未作统计学分析。4种狼疮疾病相关基因与狼疮活动指数SLEDAI无相关性。结论SLE患者TRIM,IL-4R,KLRC1和EGR1基因与正常人相比有明显差异,但与疾病活动程度无关。     

运用基因芯片初步研究类风湿关节炎患者外周血单个核细胞基因表达谱特征

目的运用基因芯片技术研究类风湿关节炎(RA)基因表达谱特征,从全基因组角度探讨RA相关致病基因。方法提取5例活动期RA患者和3名健康对照者外周血单个核细胞(PBMC)中总RNA,Illumina寡核苷酸基因芯片分析每个样本的48 000个基因变化。反转录-聚合酶链反应(RT-PCR)验证芯片结果。结果RA患者与健康对照者相比2倍上调基因122个,2倍下调基因29个。分子及生物功能分析显示上调表达的基因涉及传递蛋白、蛋白翻译合成、代谢、转录调控、信号转导、趋化因子、细胞周期和凋亡相关基因等种类。结论RA涉及多环节的病理过程,基因芯片技术有利于进一步揭示RA分子机制,发现新的治疗靶标。     

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.     

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.     

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.     

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。     

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     

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。     

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.     

治疗高血压药物的经济学评价

重视高血压治疗中的经济学评价,对利用我国有限的卫生资源来遏制高血压对人民群众的危害有着重要的现实意义。药物经济学对于药物治疗的成本和治疗的结果给予同样的关注。因为治疗高血压的费用,不仅涉及药物价格,还包括患者的危险水平,降压疗效和对临床终点事件的影响,以及治疗的依从性和安全性。因此药物经济学更强调整体成本和价-效比。低危病人,若非药价低廉,治疗的价-效比不够理想。而在高危的患者,价-效比越小越经济而不是药费越便宜越好。