两种非酒精性肝细胞脂肪变性模型的比较

目的 比较50%胎牛血清,油酸分别建立HL-7702细胞脂肪肝的离体细胞模型的优缺点.方法 采用50%胎牛血清、油酸分别建立HL-7702细胞脂肪肝的离体细胞模型,并用四甲基偶氮唑盐法检测细胞活性,油红O染色观察细胞内脂滴数量、甘油-3-磷酸氧化酶法检测细胞内甘油三酯含量,比较两种方法的优缺点.两组计量数据比较用t检验,多个样本均数比较采用方差分析.结果 0.5mmol/ml油酸组和50%胎牛血清组都能建立较好的肝细胞脂肪变性模型,其中0.5 mmol/ml油酸诱导的细胞脂变更为明显,细胞内甘油三酯含量为(893.2±89.1)μg/mg,50%胎牛血清组细胞内甘油三酯含量为(411.6±126.6)μg/mg,两组比较,F=72.288,P＜0.01,差异有统计意义.结论 0.5 mmol/ml油酸诱导的肝细胞脂肪变性模型具有实用性和可重复性.该模型为非酒精性脂肪性肝病的研究提供了一条有效、简便、可行的新途径.     

漏芦提取物对CCl_4诱导大鼠原代肝细胞损伤的保护作用及机制探讨

目的观察漏芦提取物对CCl_4诱导大鼠原代肝细胞损伤的保护作用,并探讨其机制。方法将原代培养的大鼠肝细胞分为分为正常对照组、CCl_4损伤组及漏芦大、中、小剂量组。培养24 h以后,向漏芦大、中、小剂量组分别加质量浓度为5、2.5、1mg生药/ml的漏芦提取物,向其他组加等量RPMI-1640培养液;培养6 h,向CCl_4损伤组及漏芦大、中、小剂量组加入50%CCl_4-DMSO溶液(CCl_4终浓度为5 mmol/l),正常对照组加等量RPMI 1640培养液。继续培养6 h,检测各组细胞培养上清液中的天门冬氨酸转移酶(AST)、丙氨酸转移酶(ALT)、丙二醛(MDA)、谷光苷肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)。结果与正常对照组比较,CCl_4损伤组肝细胞培养上清液中ALT、AST水平明显升高(P均<0.01),SOD、GSH-Px活性显著降低(P均<0.01);与CCl_4损伤组,漏芦大、中、小剂量组肝细胞培养上清液中ALT、AST、DA水平明显降低(P均<0.05),SOD、GSH-Px活性显著增加(P均<0.05),且里一定剂量依赖性。结论漏芦提取对物CCl_4诱导的大鼠原代肝细...     

有效原代猪肝细胞培养体系的研制

目的建立一种有效的原代猪肝细胞长期培养体系．为人工肝生物材料提供生存环境．方法把经二步胶原酶灌注法分离的猪肝细胞按３×１０５分别接种于无血清培养基、１００ｍＬ／Ｌ胎牛血清培养基、１００ｍＬ／Ｌ的猪门静脉血清培养基中．光镜下观察各体系中肝细胞形态变化过程．采用Ｂｅｃｋｍａｎ全自动生化分析仪检测各培养体系不同时间培养上清中Ａｌｂｕｍｉｎ,Ｕｒｅａ的含量．结果原代猪肝细胞在三种不同培养体系中存活时间分别是：无血清培养基４ｄ～５ｄ,胎牛血清培养基３５ｄ～３７ｄ,猪门静脉血清培养基５７ｄ～５８ｄ．存活的肝细胞功能活性除无血清培养组不明显外,胎牛血清培养组可维持４ｗｋ左右,猪门静脉血清培养组则可维持８ｗｋ左右结论门静脉血清培养体系是原代猪肝细胞较为理想的生存环境．     

红景天苷对原代培养脂肪变性大鼠肝细胞细胞色素P4502E1蛋白表达的抑制作用

目的研究红景天苷对原代培养大鼠脂肪变性肝细胞细胞色素P4502E1(CYP2E1)蛋白表达的抑制作用。方法体外分离、原代培养SD大鼠肝细胞,选取培养48 h的大鼠肝细胞,加入0.25 mmol/L棕榈酸诱导肝细胞脂肪变性,同步加入150μg/ml的红景天苷继续培养24 h,检测培养肝细胞上清液ALT、AST、丙二醛(MDA)和肝细胞甘油三酯(TG)含量的变化。采用油红O染色观察肝细胞脂滴沉积,采用蛋白质印迹法检测CYP2E1蛋白表达的变化。结果以0.25 mmol/L棕榈酸和150μg/ml红景天苷同步培养细胞24 h,红景天苷干预组细胞培养上清ALT[(11.3±1.0)U/L、AST(3.7±0.5)U/L、MDA(1.7±0.2)nmol/ml和TG(105.7±16.8)μg/mg protein]均显著低于棕榈酸模型组[ALT(13.5±1.2)U/L,P0.05;AST(5.9±0.7)U/L,P0.05;MDA(4.2±0.9)nmol/ml,P0.01;TG(268.3±25.8)μg/mg protein,P0.01];油红O染色显示红景天苷处理肝细胞脂滴明显减少,蛋白质印迹法显示红景天苷能有效抑制脂肪变性肝细胞CYP2E1蛋白表达的上调。结论红景天苷抑制细胞CYP2E1蛋白表达可能是其防治肝细胞脂肪变性的主要机制之一。     

犬恶丝虫感染期幼虫的体外培养

本文比较了犬恶丝虫在3种培养液中的发育情况。犬恶丝虫的感染期幼虫来自阳性埃及伊蚊,处理后接种于下列3种培养液中:(A)含10%人AB型血清,采用HEPES进行缓冲的RPMI 1640。(B)含有机酸(100ml内含失水苹果酸67mg、琥珀酸6mg和α-酮戊二酸37mg、糖(100ml内含葡萄糖70mg和蔗糖2668mg)和10%胎牛血清的TC-199培养液(C)含10%胎牛血清的Grace昆虫培养液。血清在加热灭活后加入培养液。培养液中加入苄青霉素和链霉素并调节pH值为7.2～7.4。以10～12条感染期幼虫培养于盛     

不同胎牛血清、培养瓶体外培养大鼠骨髓间充质干细胞效果比较

目的 比较普通、高纯度胎牛血清培养基及塑料瓶、玻璃瓶体外培养全骨髓贴壁法分离的骨髓间充质干细胞的效果.方法 采用全骨髓贴壁法分离大鼠骨髓间充质干细胞,常规传代及纯化.分为4组.其中2组分别采用含10％标准胎牛血清的DMEM培养基、含10％高纯度胎牛血清的DMEM/F12培养基培养,分别为标准胎牛血清组和高纯度胎牛血清组.2组分别用一次性塑料瓶和玻璃瓶培养,分别为塑料瓶组和玻璃瓶组.比较标准胎牛血清组合高纯度胎牛血清组、塑料瓶组和玻璃瓶组骨髓间充质肝细胞细胞的黏附、增殖、分化状况及细胞形态.结果 标准胎牛血清组贴壁细胞少,原代第10天细胞密度低,细胞透亮,出现突起,类似神经样细胞,传代后细胞呈瘢痕样及神经球样细胞形态;高纯度胎牛血清组贴壁细胞较多,密度均匀,呈典型的栅栏样、鱼群及漩涡样集落生长,传代后呈典型长梭形紧密排列.塑料瓶组塑料培养瓶内细胞贴附牢固,呈典型形态生长;玻璃瓶组玻璃培养瓶内细胞换液时部分脱落.结论 体外培养骨髓间充质肝细胞采用高纯度胎牛血清、一次性塑料瓶者培养效果好于普通胎牛血清和玻璃瓶.     

胎肝细胞培养上清液对小鼠急性肝衰竭的保护作用

为探讨胎肝细胞治疗重型肝炎的机制,将84只小鼠随机分为3组(每组28只),经D-氨基半乳糖/内毒素(D-galn/LPS)诱导急性肝衰竭后,分别给予生理盐水(NS)、RPMI1640培养液(RPMI)和人胎肝细胞培养上清液(FHCS)治疗,各组分别取14、8、6只小鼠进行存活率、血清谷丙酶(ALT)及肝脏病理观察。结果显示,NS组和RPMI组24小时存活率分别为14.3％和21.4％;ALT升高达207.2±41.3IU/L和188.4±52.6IU/L。FHCS组小鼠存活率为57.1％,ALT为92.5±28.7IU/L,与对照组相比差异显著(P<0.05,P<0.01)。此外,对照组分别有5(83.3％)只和4(66.7％)只动物肝组织病理改变在Ⅲ级以上,而FHCS组仅有2(33.3％)只。上述结果表明,FHCS对急性肝衰竭有保护作用,同时提示胎肝细胞分泌产物可能与保护肝细胞、治疗重型肝炎及肝衰竭的机制有关。     

高血糖与非肥胖糖尿病小鼠肝脏病变的关系

目的观察非肥胖糖尿病(NOD)小鼠的肝脏损伤,探讨高血糖与肝脏病变的关系。方法53只雌性NOD小鼠随机分为普通饲料组(NC组,n=23)和高脂饲料组(HF组,n=30)。所有小鼠均在确诊为糖尿病后2周或观察至9月龄时处死。检测各组的肝指数、生化指标及肝脏组织学改变。结果NC组小鼠均无肝细胞脂肪变性发生,但糖尿病组(NC-DM组)肝指数、血糖、TG及ALT、AST均高于非糖尿病组(NC-ND组)小鼠(P0.05)。HF组小鼠均出现不同程度的肝细胞脂肪变性,糖尿病组(HF-DM)重于非糖尿病组(HF-ND)(P0.01),所有小鼠均表现为肝脏轻度非特异性炎症。DM组小鼠肝细胞胞质有糖原蓄积。结论NOD小鼠肝细胞脂肪变性与高脂饮食有关,高血糖可引起胞质糖原蓄积及肝酶学异常,并加剧肝细胞脂肪变性的程度。     

大鼠结膜杯状细胞的体外培养及鉴定

目的体外分离、培养大鼠结膜杯状细胞,观察其形态、结构等生物学特征。方法取大鼠穹隆部结膜,加入含10%胎牛血清的RPMI 1640培养液中行组织块培养,5~7 d后酶消化法传代。观察原代及传代细胞的形态及生长情况,并应用特殊免疫组化染色方法分别对原代、传代培养的杯状细胞进行鉴定,用Image J软件计算细胞纯度。结果倒置相差显微镜下可见杯状细胞呈鹅卵石样,5~7 d在组织块周围形成环状结节。免疫组化结果显示,培养的原代细胞角蛋白(CK)4染色阴性,CK7染色阳性,且CK7阳性细胞数占(97.3±2.0)%。传代杯状细胞中,绝大多数凝集素UEA-I染色为阳性,阳性细胞占(91.2±2.7)%。结论大鼠穹窿部结膜可以成功培养出杯状细胞,传代细胞仍保持原代细胞特性,且纯度较高。     

胚胎大鼠神经干细胞培养方法的改进

分别以DMEM/F12、2?7、20 ng/ml的碱性成纤维细胞生长因子、20 ng/ml的表皮生长因子无血清培养液和5%胎牛血清培养液预先培养大鼠神经干细胞(NSCs)2~3 d后,换为无血清培养液;应用倒置显微镜和免疫细胞化学法对其进行观察和比较。结果用含胎牛血清培养液培养的NSCs聚球速度较无血清培养液明显加快,可提前3~4 d观察到NSCs球。细胞染色证实细胞系巢蛋白阳性,经血清培养液诱导分化后部分呈抗神经元特异性烯醇化酶(NSE)阳性,部分呈抗胶质纤维酸性蛋白阳性,表明可分化为神经元和胶质细胞。认为含血清培养液预先培养可以加快细胞培养速度,是获得NSCs的经济有效方法。     

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。