罗格列酮对肝星状细胞增殖与胶原合成的影响

目的 观察罗格列酮对肝星状细胞增殖与Ⅰ型胶原合成的影响.方法 采用HSC-T6肝星状细胞系,将培养的肝星状细胞(HSC)分为对照组、不同浓度的罗格列酮组(5、10、20 μmol/L).采用MTT法观察罗格列酮对HSC增殖的影响,采用ELISA法检测细胞培养液上清中Ⅰ型胶原的含量.结果 与对照组相比,在10、20μmol/L罗格列酮组,罗格列酮可以抑制HSC的增殖(P<0.05),同时细胞堵养液上清中Ⅰ型胶原含量分别为135.72±6.00 ng/ml、128.65±5.34 ng/ml,均低于对照组(149.35±6.65 ng/ml),差异有统计学意义(P<0.05).结论 罗格列酮可以抑制HSC的增殖,抑制肝星状细胞Ⅰ型胶原的合成.     

PPARγ表达上调体外抑制肝星状细胞的增殖

目的:观察过氧化物酶体增殖物激活受体γ(PPARγ)表达上调对肝星状细胞增殖及凋亡的影响.方法:体外培养HSC-T6细胞,取对数生长期的细胞,应用1,2,3μmo/L不同浓度的PPARγ激动剂15d-PGJ2作用24 h.同时以不加药物只加无血清培养基作为对照组,24 h后应用RT-PCR的方法观察分析各组间肝星状细胞PPARγmRNA表达的变化,MTT检测HSC增殖.流式细胞仪分析HSC-T6细胞凋亡.结果:经1,2,3μmo/L 15d-PGJ2处理的HSC-6细胞中PPARγmRNA表达比例上调(0.513±0.031,0.697±0.018,0.674±0.03 2 vs 0.198±0.021).MTT结果显示不同浓度的15d-PGJ2对HSC-T6细胞的增殖均有抑制作用,以2μmol/L组最为显著(54.6%±11.1%).流式细胞仪结果显示15d-PGJ2处理组凋亡细胞数明显增多.结论:PPARγ表达上调能够抑制HSC-T6增殖并诱导其凋亡.     

羟基喜树碱对HSC-T6细胞增殖与凋亡的影响

目的 探讨羟基喜树碱(HCPT)对体外培养大鼠肝星状细胞(HSC)的增殖与凋亡的影响.方法 大鼠肝星状细胞(HSC-T6)和大鼠正常肝细胞(BRL-3A)分别在实验组(分别以含HCPT浓度为0.008、0.016、0.031、0.063、0.125,0.25、0.5、1、2、4、8、16、32mg/L的培养液培养)和对照组(单纯培养)体外培养24 h.用四甲基偶氮唑盐法检测细胞增殖情况,找出HCPT对HSC-T6细胞增殖抑制的最佳作用浓度;流式细胞仪检测细胞凋亡率;透射电子显微镜下观察细胞凋亡的形态学变化情况;琼脂糖凝胶电泳法检测DNA片段化.多个样本均数的比较采用单因素方差分析,两样本均数的比较采用t检验.结果 HCPT对HSC-T6细胞和BRL-3A细胞的增殖抑制率随着药物浓度的升高而逐渐升高;当HCPT浓度＞0.5mg/L时,对BRL-3A细胞的毒性作用显著增高(P值均<0.05);0.5 mg/L对HSC-T6细胞增殖抑制作用最大,为HCPT对HSC-T6细胞增殖的最佳抑制浓度.0.125、0.25、0.5 mg/L的HCPT作用HSC-T6细胞24h,流式细胞仪检测显示细胞凋亡率分别为13.46%±2.42%、26.25%±5.65%、47.05%±8.76%,与对照组(4.89%±1.80%)相比,差异有统计学意义(F=34.24,P<0.01).0.5mg/L的HCPT作用HSC-T6细胞24 h,透射电子显微镜下可见细胞体积缩小,核仁消失,染色质浓缩聚集成团块状,沿核膜排列等凋亡形态学改变;琼脂糖凝胶电泳可见明显的DNA梯度带形成.结论 HCPT在体外可以明显地抑制HSC-T6细胞的增殖、诱导HSC-T6细胞凋亡,作用强度呈剂量依赖性.     

抗纤软肝颗粒对大鼠肝星状细胞增殖的影响

目的:探讨抗纤软肝颗粒(KXRG)对大鼠肝星状细胞(HSC)增殖的影响。方法:传代培养的HSC-T6(肝星状细胞系)与KXRG(1.25mg/ml、2.5mg/ml、5mg/ml)及其药物血清(5%、10%、20%)共同培养72小时后,应用MTT法测定细胞增殖、流式细胞仪测定HSC细胞周期各时相的DNA含量。结果:KXRG及药物血清均能显著抑制HSC的增殖,使细胞周期阻滞于S期,且2.5mg/ml药物和10%药物血清作用最强(P0.01)。结论:KXRG通过抑制HSC的活化,阻滞其细胞周期的正常进行,进而抑制HSC增殖,从而起到抗肝纤维化的作用。     

LY294002在外源性H2S预处理肝纤维化大鼠肝星状细胞增殖、凋亡中的作用

目的探讨硫化氢(hydrogen sulfide,H2S)通过PI3K/Akt信号通路对大鼠肝星状细胞增殖、凋亡的调节作用。方法 MTT法分别检测NaHS(H2S的供体)和PI3K/Akt信号通路的特异性抑制剂LY294002对大鼠肝星状细胞(HSC-T6)的增殖、增殖抑制率的影响;采用流式细胞技术检测药物的有效浓度对HSC-T6凋亡的调节。结果低浓度(50μmol/L)的NaHS能够明显促进HSC-T6的增殖(F=955.949,P=0.000),LY294002可抑制HSC-T6的增殖,伴随药物浓度的升高细胞存活率降低(P<0.05),并诱导HSC-T6的凋亡,而二者联合应用诱导HSC-T6凋亡作用增强。结论 H2S可能通过PI3K/Akt信号通路促进HSC-T6的增殖,但对肝星状细胞的凋亡抑制作用并不显著,H2S可协同LY294002共同诱导HSC-T6细胞的凋亡。     

大黄素对HSC-T6细胞增殖及细胞周期影响的研究

目的 研究大黄素干预对大鼠肝星状细胞株(HSC-T6)增殖和细胞周期的影响，探讨其抗肝纤维化的具体机制。方法 应用大鼠肝星状细胞株伪传代培养，采用不同浓度大黄素进行干预，观察各组细胞在形态学、增殖程度、细胞周期、增殖细胞核抗原(PCNA)表达等多方面的变化。结果 ①5-15Mg／L浓度大黄素干预24h后，对T6细胞的增殖产生显著的抑制效应，同时免疫组化显示PCNA标记指数随着药物浓度的增高其表达阳性率反而逐渐降低。②大黄素同时能够对T6细胞的细胞周期产生阻滞作用，随着浓度的增高，C0／G1期细胞比例逐渐上升。结论 大黄素在体外对于HSC-T6的增殖具有明显的抑制作用，而这种作用可能是通过改变了T6细胞正常的细胞周期来实现的。     

抗纤软肝颗粒对肝星状细胞凋亡的影响

探讨抗纤软肝颗粒对肝星状细胞(HSC)凋亡的影响.方法:传代培养的大鼠肝星状细胞系HSC-T6与中药复方抗纤软肝颗粒(终浓度为5mg/ml、2.5mg/ml、1.25mg/ml)及药物血清(5%、10%、20%)共同培养48小时后,应用TUNEL法及流式细胞仪测定细胞凋亡、免疫组化检测Bcl-2/Bax.结果:TUNEL法及流式细胞仪测定结果显示抗纤软肝颗粒药物及含药血清均可诱导HSC凋亡(P＜0.05);并能下调凋亡抑制分子Bcl-2,上调促凋亡分子Bax(P＜0.01).结论:抗纤软肝颗粒可下调Bcl-2/Bax比率,以增加细胞对凋亡的敏感性,从而促进HSC凋亡.     

神经生长因子对肝星状细胞生物行为学的影响

目的 探究神经生长因子(NGF)对肝星状细胞(HSCs)增殖、凋亡的影响及其可能的作用机制.方法 对数生长期HSC-T6细胞随机分为对照组(0 ng/ml)、NGF低剂量组(100 ng/ml)、NGF中剂量组(200 ng/ml)和NGF高剂量组(300 ng/ml),采用CCK-8法检测HSC-T6的增殖情况;采用流式细胞仪检测HSC-T6的凋亡情况;采用逆转录-聚合酶链反应(RT-PCR)检测α-平滑肌肌动蛋白(SMA)、Ⅰ型胶原及基质金属蛋白酶组织抑制因子(TIMP)-1的mRNA表达水平;采用Western印迹检测含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-3、p53的蛋白表达变化.结果 与对照组比较,NGF低、中、高剂量组的细胞增殖均受到显著抑制,细胞凋亡率则显著增加,且各NGF处理组之间的这种变化呈明显的剂量依赖效应(P<0.05).NGF低、中、高剂量组细胞中α-SMA、Ⅰ型胶原及TIMP-1的mRNA相对表达水平较对照组均显著下调,并且该变化趋势随着NGF剂量的增加而增强(P<0.05).与对照组比较,NGF低、中、高剂量组细胞中Caspase-3、p53的表达水平均显著增加,各NGF剂量组之间的蛋白表达变化具有明显的剂量依赖性(P<0.05).结论 NGF通过调控TIMP-1、Ⅰ型胶原、Caspase-3和p53等基因的表达抑制HSCs的增殖并诱导其凋亡,这可能为肝纤维化的临床治疗开辟新的思路.     

白花丹对大鼠肝星状细胞增殖、凋亡及细胞周期的影响

目的: 观察白花丹含药血清对大鼠肝星状细胞(HSC-T6)增殖、凋亡细胞周期的影响,探讨白花丹抗肝纤维化的作用及其机制.方法: SD大鼠用白花丹水煎液灌胃(大、中、小剂量),取得含药血清.含药血清按不同浓度(50、100、200 mL/L)分组与大鼠HSC-T6孵育.设立空白对照组,并以秋水仙碱和复方鳖甲软肝片含药血清为阳性对照组.孵育后各组采用MTT比色法检测各组HSC-T6的增殖;流式细胞仪检测细胞凋亡及各细胞周期DNA含量的百分比.结果: HSC-T6白花丹含药血清各组与空白组比较,增殖抑制率和细胞凋亡率明显增高,有显著性差异( P＜0.01);并且含药血清浓度增高时,HSC-T6的抑制率、凋亡率也逐渐增强;血清高浓度组的增殖抑制率和细胞凋亡率明显高于秋水仙碱组( P＜0.01),与复方鳖甲软肝片组相当;各组的G2/M期细胞百分比无明显变化.HSC-T6白花丹含药血清各组与空白组比较,G0/G1期的细胞百分比明显增高(55.23%±2.83%,52.60%±2.26%,48.75%±1.37%vs 44.08%±1.41%,均P＜0.01)、S期细胞百分比明显降低(31.47%±1.26%,34.14%±1.17%,40.28%±1.62% vs 47.36%±1.35%,均P＜0.01).并且含药血清浓度下降时,G0/G1期的细胞百分比逐渐下降,S期细胞百分比逐渐增高.同一血清浓度下,与秋水仙碱组比较,白花丹组的G0/G1期细胞百分比明显较高( P＜0.01),S期细胞百分比明显较低;而与复方鳖甲软肝片组无显著性差异.结论: 白花丹含药血清可以抑制HSC-T6增殖,诱导其凋亡,且作用具有剂量依赖性.白花丹含药血清抑制HSC-T6增殖的机制可能是使HSC-T6细胞周期停滞于G0/G1期,阻止其通过G1/S关卡.     

大黄蔗虫丸对大鼠肝星状细胞活化及增殖的影响

目的观察大黄蔗虫丸对大鼠肝星状细胞活化及增殖的影响。方法用链酶蛋白酶和胶原酶原位灌注消化正常大鼠肝脏,Nycodenz密度梯度离心分离肝星状细胞(HSC),在制备大黄蔗虫丸大鼠药物血清的基础上,温育HSC。MTT比色法测定细胞增殖,流式细胞仪检测细胞凋亡及增殖周期情况。结果药物血清对体外培养的HSC有抑制作用,且存在浓度剂量依赖关系,但作用较弱,需在20%以上方见到显著性差异(P<0.05),对HSC凋亡及细胞增殖周期则无显著影响(P>0.05)。结论大黄蔗虫丸对HSC增殖有一定抑制作用,但可能不是其抗肝纤维化作用的主要途径,其抗肝纤维化作用与诱导细胞凋亡无关。     

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。     

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.     

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.