血管活性肠肽对体外大肠癌HT29细胞粘附和侵袭作用的影响

目的 观察血管活性肠肽(VIP)和双丁酰环磷酸腺苷(db-cAMP)对体外大肠癌HT29细胞对大鼠血管内皮的粘附和侵袭的影响。方法 取出大鼠主动脉，建立体外癌细胞-血管内皮粘附试验模型．用光镜和扫描电镜观察HT29细胞对大鼠血管内皮粘附作用和其形态学变化．以及VIP和db-cAMP对细胞粘附的影响并托扫描电镜上计数量化分析。结果 在血管内皮上可清晰辨认痛细胞并可观察其对血管内皮的粘附和侵袭的形态学改变．VIP组的粘附细胞和高活性粘附细胞显多于对照组(P<0.05)．相反db-cAMP则无此作用．结论 VIP可促进体外HT29细胞对血管内皮的粘附和侵袭，其机制可能与激活癌细胞A激酶系统有关．     

整合素ανβ6对结肠癌细胞胞外基质降解的调控作用

目的 探讨整合素ανβ6在结肠癌细胞侵袭中的作用,并揭示癌细胞调控胞外基质降解的相关分子机制.方法 利用质粒构建ανβ6特异性siRNA表达载体,细胞侵袭实验检测沉默ανβ6对结肠癌细胞HT29体外侵袭能力的影响;分别通过Western blot实验、明胶酶谱实验检测沉默ανβ6对结肠癌细胞胞外信号调节激酶(ERK)、尿激酶型纤溶酶原激活物(uPA)及基质金属蛋白酶(MMP-2、MMP-9)表达的影响;[3H]标记的Ⅳ型胶原降解分析检测沉默ανβ6对结肠癌细胞胞外基质降解能力的影响.结果 特异性siRNA表达载体可有效抑制HT29细胞中ανβ6的表达和结肠癌细胞的体外侵袭能力,显著抑制结肠癌细胞ERK1/2的表达,并抑制其活化形式磷酸化ERK1/2的表达;沉默ανβ6表达显著抑制结肠癌细胞uPA,pro-MMP-9和pro-MMP-2的分泌,以及丝裂原活化蛋白激酶(MAPK)依赖的胞外基质降解.结论 整合素ανβ6通过MAPK信号通路调控uPA、MMP-9和MMP-2的分泌及活性,从而调控细胞外基质的降解,在结肠癌侵袭转移中发挥了重要作用.     

吲哚美辛对受损颈动脉再内皮化的作用及机制

目的研究吲哚美辛对受损颈动脉再内皮化的作用及机制。方法在SD大鼠体内通过球囊法制备颈动脉损伤模型,腹腔注射吲哚美辛,观察其对血管内皮修复的影响;密度梯度离心法获得大鼠单核细胞,应用EGM-2培养基体外诱导培养获得内皮祖细胞(EPCs)。采用不同浓度的吲哚美辛处理晚期EPCs24 h后,MTT法检测细胞的增殖活性,细胞差异贴壁法检测EPCs的粘附能力,改良的Boyden小室检测EPCs的迁移能力,Matrigel基质胶检测EPCs的成血管能力。结果吲哚美辛可明显促进损伤血管内皮的修复,其机制可能与吲哚美辛增强EPCs的增殖、粘附、迁移及成血管能力有关。结论吲哚美辛促进了损伤血管内皮的再内皮化进程。     

麝黄消瘤方抑制肝癌细胞侵袭粘附作用的实验研究

目的：观察麝黄消瘤方（SHF）兔含药血清对SMMC-7721肝癌细胞侵袭粘附能力的影响及其机制的探讨。方法：以体外培养的肝癌细胞株为研究对象，将肝癌细胞株分别接种于中药组、对照组血清中培养，绘制细胞生长曲线，计算细胞生长抑制率。以流式细胞仪（FCM）测SHF对肝癌细胞表面细胞间粘附分子-1（ICAM-1）表达的影响。以免疫组化法观察SHF对肝癌细胞抑癌基因nm23-H1蛋白水平的表达。以细胞侵袭实验观察SHF对与纤维连接蛋白（FN）粘附的SMMC-7721细胞侵袭运动能力及其生长状况。以岍法测量SHF对肝癌细胞与FN粘附的影响。结果：中药组血清较对照组明显抑制肝癌细胞的体外生长，在第5天抑制率最高达66．25％；中药处理的肝癌细胞ICAM-1表达明显低于对照组，nm23-H1蛋白水平明显高于对照组。SHF可抑制肝癌细胞的侵袭运动能力及其与FN之间的粘附。结论：SHF可抑制SMMC-7721细胞的侵袭粘附能力，其作用与中药血清上调nm23-H1蛋白水平和降低ICAM-1表达等有关。     

基质细胞衍生因子/趋化因子受体4轴参与胰腺癌细胞侵袭的体外实验研究

目的 探讨胰腺癌细胞株趋化因子受体(CXCR)4的表达及其与增殖、侵袭和粘附的关系.方法 采用RT-PCR检测3个胰腺癌细胞株中CXCR4 mRNA的表达;Western印迹法检测胰腺癌细胞株中CXCR4的蛋白表达;共聚焦显微镜检测基质细胞衍生因子(SDF)-1α作用下AsPC-1细胞内钙荧光强度变化;MTT法检测细胞增殖;细胞体外粘附试验及Transwell体外侵袭实验观察胰腺癌细胞的粘附及侵袭能力.结果 3个胰腺癌细胞株均不同程度地存在CXCR4 mRNA及蛋白表达,AsPC-1细胞表达最强,而SW1990表达最弱;CXCR4功能性表达于AsPC-1细胞;SDF-1α不同程度的促进3种胰腺癌细胞的增殖、侵袭及粘附,尤以AsPC-1细胞最为明显,而SW1990细胞最弱;AMD3100可抑制SDF-1α所致的促增殖、侵袭及粘附作用.结论 胰腺癌细胞表达CXCR4mRNA及蛋白,SDF-1α通过与CXCR4作用影响胰腺癌细胞的侵袭转移,该效应与CXCR4表达程度密切相关.     

CTHRC1对结直肠癌细胞增殖、迁移和侵袭的影响

目的观察胶原三股螺旋重复蛋白(CTHRC)1对结直肠癌HT29细胞增殖、迁移和侵袭的影响。方法采用RT-PCR和免疫组织化学方法检测结直肠癌组织中CTHRC1的表达情况。采用电穿孔转染法,将pc DNA3.1-CTHRC1和CTHRC1 si RNA转染HT29细胞中,通过克隆形成实验检测CTHRC1对HT29细胞增殖的影响;通过伤口愈合实验和Boyden小室检测CTHRC1对HT29细胞迁移和侵袭能力的影响。结果 RT-PCR和免疫组织化学检测显示,CTHRC1在结直肠癌组织中的表达显著高于正常对照组织。增殖、迁移和侵袭实验结果显示,对照组、转染CTHRC1和转染CTHRC1 si RNA的结直肠癌HT29细胞克隆形成数分别为(147.34±15.86)、(234.17±27.86)和(66.03±8.95);伤口宽度百分比分别为(1.00±0.00)、(0.37±0.12)和(1.48±0.35);发生侵袭的细胞数分别为(201.24±33.54)、(415.36±45.63)和(96.13±15.46);转染CTHRC1可显著促进结直肠癌细胞的增殖、迁移和侵袭能力;而转染si-CTHRC1可显著抑制结直肠癌细胞的增殖、迁移和侵袭能力。结论 CTHRC1可明显增加HT29细胞增殖、迁移和侵袭能力,这可能是CTHRC1参与结直肠癌发生发展的机制。     

miR-301a促进人结肠癌细胞株HT29细胞侵袭与转移的机制

目的探讨miR-301a对结肠癌侵袭转移的影响及分子机制。方法实时荧光定量PCR检测40例结肠癌患者癌组织中miR-301a的表达水平,探究结肠癌和miR-301a表达的关系。同时在人结肠癌细胞株HT29细胞过表达和干扰miR-301a,MTT法检测miR-301a对HT29细胞增殖的影响。蛋白免疫技术和实时荧光定量PCR在分子水平上检测miR-301a表达对HT29细胞中侵袭相关分子的表达水平,探究miR-301a调控下的转化生长因子(TGF)-β/Smad4/基质金属蛋白酶(MMP)-9信号通路对结肠癌侵袭转移的影响。结果实时荧光定量PCR结果显示40例结肠癌患者癌组织中miR-301a的表达水平明显升高,且随着肿瘤分期分级的升高,其水平明显升高。MTT法检测发现过表达miR-301a的HT29细胞数量明显增加,而干扰miR-301a则细胞数目减少,表明miR-301a可促进结肠癌细胞的增殖。在分子水平上蛋白免疫技术和实时荧光定量PCR检测结果显示miR-301a通过下调TGF-β和Smad-4的表达,促进MMP-9的表达,最终促进癌细胞的侵袭转移。结论 miR-301a通过下调TGF-β/Smad4信号通路,促进MMP-9的表达,最终促进结肠癌的侵袭转移。     

五味子多糖对人结肠癌HT29细胞增殖的抑制作用及其机制

目的探讨五味子多糖含药血清对人结肠癌细胞株HT29体外增殖的影响及作用机制。方法采用人结肠癌细胞株HT29作为研究对象,MTT法检测不同浓度的五味子多糖含药血清对细胞增殖的抑制作用;倒置显微镜和荧光显微镜观察细胞形态和细胞核的改变;流式细胞仪分析细胞凋亡的情况。结果五味子多糖含药血清对HT29细胞的增殖有抑制作用而且呈剂量和时间依赖性。五味子多糖含药血清作用72 h后,荧光显微镜可见凋亡细胞及凋亡小体;流式细胞仪检测凋亡。结论五味子多糖含药血清对HT29结肠癌细胞的增殖有抑制作用,其作用机制与诱导细胞凋亡有关。     

大肠癌癌细胞缝隙连接蛋白Cx43表达及与肿瘤转移的相关性探讨

目的探讨癌细胞能穿出血管内皮进入靶器官继续生长形成转移瘤的机理。方法将人血管内皮细胞单独培养及分别与大肠癌低转移细胞HT29、高转移细胞Lovo共同培养,至细胞长成单层采用免疫组化SP法检测细胞缝隙连接蛋白Cx43表达。结果在单纯血管内皮细胞Cx43蛋白表达最强,次之为与HT29细胞共培养,而与Lovo细胞共培养后Cx43几乎无阳性表达。结论 Cx43蛋白在大肠癌细胞转移中起重要作用;检测Cx43蛋白表达有助于判断肿瘤的转移能力。     

蛴螬提取物对人肺癌A549细胞诱导凋亡作用的形态学观察

目的 探讨蛴螬提取物对人肺癌A549细胞诱导凋亡的形态学改变.方法 应用MTT法、HE染色、扫描电镜的方法 观察蛴螬提取物对体外培养的人肺癌A549细胞形态学影响.结果 蛴螬石油醚提取物对人肺癌A549细胞株的增殖抑制作用呈剂量依赖性和时间依赖性(P＜0.05或P＜0.01).倒置显微镜下蛴螬石油醚提取物80 μg/ml作用后,早期细胞出现凋亡形态学变化,晚期细胞发生膜破裂坏死.HE染色和扫描电镜观察细胞出现典型凋亡形态变化.结论 蛴螬提取物在体外对人A549肺癌细胞株有显著性抑制增殖和诱导凋亡作用.     

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。     

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。     

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。     

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.