基因表达谱芯片分析日本血吸虫感染慢性期CD4+细胞的应答特征

目的　立足于全基因水平考察日本血吸虫慢性感染小鼠CD4+细胞的相关细胞因子的变化 ,探讨其与疾病的相关性。方法　采用高密度寡核苷酸芯片 (Affemetrix芯片 )结合三色流式细胞术 ,对日本血吸虫感染 13周的小鼠脾脏中的CD4+细胞进行基因转录及蛋白表达水平的分析 ,获得相关细胞因子的变化图谱。结果　日本血吸虫慢性感染小鼠CD4+细胞中IL 4的转录及蛋白表达水平均明显高于正常小鼠 (P <0 .0 5) ,尤其以可溶性虫卵抗原刺激后最为显著 (P <0 .0 1) ;感染小鼠IFN γ的转录及蛋白表达水平也有所增高 ,但不及IL 4的升高程度 ;IL 10、IL 1β、小诱导细胞因子(smallinduciblecytokine)等因日本血吸虫的感染而升高 ,但TGF β1呈下降趋势。结论　日本血吸虫慢性感染小鼠细胞因子分泌谱表现为Th1和Th2型细胞因子并存的格局 ,但以Th2应答为主。它们可能通过诱导产生相应的趋化因子共同参与疾病的病理改变     

日本血吸虫感染过程中抑制性因子的表达特征

目的立足于基因组水平考察日本血吸虫慢性感染小鼠CD_4~+ T应答中抑制性因素的参与及可能的调控作用。方法采用高密度寡核苷酸芯片(Affymetrix芯片)对日本血吸虫感染0、3、6、13周小鼠脾脏中的CD_4~+ T细胞进行全基因组分析,获得相关抑制性因子的基因表达谱。结果基因芯片结果显示日本血吸虫感染从急性期至慢性期过程中抑制性因子的表达基因表达逐步上调,呈现基本一致的变化,包括抑制性细胞因子、阻断抗体、细胞凋亡、负调控分子的高表达等。结论从胞内信号转导通路至细胞凋亡的整体水平,多层次因素参与了日本血吸虫感染宿主免疫应答的负调控机制。     

趋化因子在日本血吸虫感染模型中的表达特征

目的 研究日本血吸虫感染模型中趋化因子(chemokine)在CD4^ T细胞中的表达概貌，并探讨其与Th细胞极化和免疫病理的相关性。方法 采用高密度寡核苷酸芯片(Affymetrix芯片)技术，从基因水平获得日本血吸虫不同感染时期CD4^ T细胞表达的趋化因子概貌，并用反转录PCR(RT—PCR)凝胶成像半定量技术验证嗜酸性粒细胞趋化因子(ECF—L)的表达水平。同时观察趋化因子表达与病理的的关联。结果 日本血吸虫虫卵沉积前，MIG、RANTES明显增强；感染6周IP-10、MIP-2、MIP-1α、MIP-1β、ECF—L升高，持续到感染13周；GRO、Lymphotactin、MCP-1、RANTES在感染13周显著增强。RT-PCR半定量验证ECF-L表达模式与基因芯片结论一致。结论 日本血吸虫人工感染小鼠的CD4^ T细胞表达大量趋化因子，它们共同参与Th1／Th2免疫应答，影响免疫病理状态；改变趋化因子格局可能影响疾病的自然转归。     

干扰素-γ对感染日本血吸虫小鼠GTP酶表达的影响

目的　探讨干扰素 γ(IFN-γ)对小鼠抗日本血吸虫感染保护力的机制。　方法　采用高密度寡核苷酸芯片 (Affymetrix芯片 )结合半定量逆转录 聚合酶链反应 (RT-PCR )方法 ,对BALB/c小鼠感染日本血吸虫过程中脾脏CD4+ T细胞进行基因转录水平分析 ,获得IFN γ诱导鸟苷三磷酸 (GTP)酶家族成员的变化图谱 ;并对其中IFN γ诱导GTP酶 (IGTP)进行分子克隆和序列测定。　结果　小鼠自然感染日本血吸虫过程中 ,IFN γ诱导GTP酶家族成员的基因表达呈现特征性变化 ,感染后 3wk ,基因表达上调或变化不显著 ;感染后 6wk至 13wk ,基因表达持续受到抑制。这种变化特征经RT PCR方法所证实。从小鼠脾脏可扩增出IGTP全长基因 ,但退火温度降低时出现IGTP基因转录缺失。　结论　小鼠急性感染日本血吸虫后 ,体内IFN-γ通路逐步受到抑制 ,针对血吸虫感染的依赖IFN-γ的抗感染保护力下降。     

日本血吸虫感染过程中宿主CD^＋4CD^＋8T细胞凋亡的相关基因转录水平的研究

目的：探讨在日本血吸虫感染的过程中,宿主CD^ 4和CD^ 8T细胞凋亡的相关基因p53,bcl-2,TGFβ在转录水平上的变化,方法：用地高辛标记的探针作原位杂交,观察凋亡相关基因在日本血吸虫感染过程中CD ^ 4和CD^ 8T细胞中转录水平的变化,结果：在日本血吸虫感染过程中,CD^ 4和CD^ 8T细胞中促凋亡基因p53和TGF－βRNA的表达随感染时间的延长而升高,感染后第10周起CD^ 4T 细胞p53和TGF－βmRNA的表达明显高于CD^ 8T细胞,差异有显著意义,抑制凋亡发生的基因bcl-2mRNA表达水平则与之相反,CD^ 4T细胞bcl-2mRNA的表达从感染从第10周起显著下降,而CD^ 4T细胞bcl-2mRNA的表达水平无显著性变化,结论：在日本血吸虫感染过程中,CD^ 4T细胞凋亡比率超过CD^ 4/CD^ 8T细胞,CD^ 4/CD^ 8T细胞比值下降与p53,bcl-2,TGF-β基因的活化有关。     

感染日本血吸虫小鼠CD_4~ 和CD_8~ T细胞中Fas及FasL的表达研究

目的　探讨在日本血吸虫感染过程中 ,宿主 CD4 和 CD8 T细胞中死亡受体 (Fas)及死亡配体 (Fas ligand,FasL)的表达情况。　方法　利用免疫组化的方法检测在日本血吸虫感染过程中 ,Fas和 Fas L 在 CD4 和 CD8 T细胞中的表达。　结果　在日本血吸虫感染过程中 ,Fas和 Fas L在 CD4 和 CD8 T细胞的表达随感染时间的延长而升高 ,但感染10 wk后 ,Fas L在 CD4 T细胞的表达接近并超过 Fas的表达。　结论　在日本血吸虫感染过程中 ,CD4 T细胞凋亡比率超过 CD8 T细胞和 CD4 / CD8 T细胞比值下降与相关蛋白 Fas、Fas L的表达有关     

多房棘球蚴感染宿主CD4~+T淋巴细胞缺失机制的探讨

目的探讨多房棘球蚴感染宿主CD4~+T淋巴细胞缺失的机制,多房棘球蚴感染与宿主T淋巴细胞亚群凋亡相互关系。方法利用尼龙柱和补体法从多房棘球蚴感染12wk,25wk和正常对照组BALB/c小鼠脾脏分离出纯CD4~+、CDS8~+T细胞,在体外分别经多房棘球蚴抗原(EmAg)、抗CD3抗原(anti-CD3)、白细胞介素-2(IL-2)、肿瘤坏死因子α(TNFα)及商陆丝裂原(PWM)刺激培养16h,用DNA凝胶电泳分析;透射电镜观察形态学变化;原位末端标记法(Tunel)和碘化丙锭(PI)双染后,流式细胞仪(FCM)分析凋亡细胞数和凋亡发生的细胞周期。结果感染25wk小鼠,CD4~+T细胞DNA电泳图均出现典型“梯形”条带,透射电镜见染色质浓染、胞质出泡、凋亡小体形成;CD8~+T细胞DNA电泳国无梯形带,透射电镜见染色质浓染,电子密度增强。FCM分析,感染12wk小鼠,CD4~+、CD8~+T细胞凋亡数与正常对照组差异无显著性意义(P>0.05);感染25wk小鼠,CD4~+T细胞凋亡数较正常对照组显著增高(P<0.01),也显著高于同组CD8~+T细胞(P<0.01)。凋亡主要发生在细胞周期S期。结论多房棘球蚴寄生宿主后期,可诱导宿主成熟CD4~+T细胞发生活化诱导性死亡(AICD),使宿主处于免疫抑制状态。     

ICOS信号与日本血吸虫感染小鼠Th9细胞极化的关系

目的 检测可诱导共刺激分子（ICOS）在日本血吸虫感染小鼠脾脏Th9细胞中的表达水平,探讨ICOS信号与Th9细胞极化之间的关系。方法 以日本血吸虫尾蚴感染小鼠,分别在感染后0、4、7、9周和12周,应用流式细胞术检测IL?9+细胞在CD4+ T细胞中的比例,以及ICOS+细胞在Th9细胞中的比例。结果 与0周比较,感染后4、7、9、12周,小鼠CD4+ T细胞中Th9细胞的比例显著增高（P < 0.05）,并且Th9细胞ICOS的表达也明显上调（P < 0.05）。结论 在日本血吸虫感染中,ICOS信号对Th9细胞的极化可能具有调控作用。     

泡球蚴感染过程中宿主CD+4细胞凋亡和凋亡相关基因转录水平的研究

目的　为了探讨泡球蚴感染宿主CD+ 4 淋巴细胞缺失的机制 ,以及泡球蚴感染与宿主CD+ 4 淋巴细胞凋亡及凋亡相关基因转录水平的相关性。方法　利用尼龙柱和补体法从泡球蚴感染 12周 ,2 5周和正常对照组BALB/c小鼠脾脏分离出纯CD+ 4 ,CD+ 8细胞 ,在体外分别经EmAg ,anti-CD3 ,IL - 2 ,TNFα ,PWM刺激培养 16h ;Tunel和PI双染后 ,FCM分析凋亡细胞数 ;并用C -myc ,TGF - β ,bcl- 2cDNA探针检测感染 2 5周EmAg诱导CD+ 4 细胞的凋亡相关基因转录水平。结果　FCM分析 ,感染 12周组CD+ 4 ,CD+ 8细胞凋亡数同正常对照组无明显差别 (P >0 0 5 ) ,感染 2 5周组CD+ 4 细胞凋亡数较正常对照组显著增高 (P <0 0 1) ,也显著性高于同组CD+ 8细胞 (P <0 0 1)。CD+ 4 细胞内C -myc,TGF - βmRNA水平较正常对照组显著升高 (P <0 0 5 ) ,而bcl- 2mRNA水平则减弱 (P <0 0 5 )。结论　泡球蚴寄生宿主后期 ,可诱导宿主成熟T细胞中CD+ 4 细胞发生凋亡 ,使宿主处于免疫抑制状态。凋亡的形成与凋亡信号增加 ,抑制信号减弱所引起。     

感染日本血吸虫小鼠CD4＋和CD8＋T细胞中Fas及Fas的表达研究

目的：探讨在日本血吸虫感染过程中，宿主CD4＋和CD8＋T细胞中死亡受体（Fas) 及死亡配体（Fas ligand,FasL)的表达情况，。方法：利用免疫组化的方法检测在日本血吸虫感染过程中，Fas和FasL在CD4＋和CD8＋T细胞中的表达，结果：在日本血吸虫感染过程中，Fas和FasL在CD4＋和CD8＋T细胞的表达随感染时间的延长而升高，但感染10wk后，FasF 在CD4＋T细胞的表达接近并超过Fas的表达，结论：在日本血吸虫感染过程中，CD4＋ T细胞凋亡比率超过CD8＋T细胞和CD4＋／CD9＋T细胞比值下降与相关蛋白Fas,FasL的表达有关。     

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.