白藜芦醇的小分子类似物RSVA314对白血病K562细胞迁移和增殖的影响

目的:探讨白藜芦醇的小分子类似物(RSVA314)对白血病K562细胞迁移和增殖的影响。方法:体外培养K562细胞,并分为4组:生理盐水对照组、RSVA314低剂量组(1μM)、RSVA314中剂量组(5μM)、RSVA314高剂量组(25μM)。采用CCK-8方法分别检测4组中K562细胞的增殖活性;采用TransweⅡ迁移实验分别检测4组K562细胞的迁移能力;采用Western blot分别检测4组K562细胞蛋白激酶B(AKT)的表达。结果:与对照组比较,RSVA314中、高剂量组OD值明显增加(P0.05或P0.01),提示其可抑制K562细胞的增殖活性。RSVA314低、中、高剂量组穿膜细胞数量明显减少,与对照组比较有明显差异(P0.05或P0.01)。Western blot检测显示RSVA314中、高浓度组可抑制AKT磷酸化的表达,与对照组比较有显著差异(P0.05或P0.01)。结论:RSVA314能抑制白血病K562细胞迁移和增殖,其机制可能与抑制K562细胞AKT磷酸化表达有关。     

蛇床子素通过抑制PI3K/AKT信号通路诱导K562细胞凋亡和增殖抑制

目的:研究蛇床子素对人慢性髓系白血病细胞K562增殖和凋亡的影响,并探讨其相关作用机制。方法:培养髓系白血病细胞K562,分别用不同浓度的蛇床子素处理(0、5、10、20μg/ml)。细胞培养48h后,利用MTT法检测K562细胞增殖;流式检测K562细胞凋亡;Western blot法检测PI3K、p-AKT、AKT、Bax、Bcl-2和Cleavage-Caspase3表达。结果:蛇床子素可以呈浓度依赖性的诱导K562细胞增殖抑制和凋亡,上调Bax和Cleavage-Caspase3表达,下调PI3K、p-AKT和Bcl-2,对AKT表达无明显影响。结论:蛇床子素可通过抑制PI3K/AKT信号通路而诱导K562细胞增殖抑制和凋亡。     

白藜芦醇对白血病K562细胞增殖、凋亡的影响及机制探讨

目的 探讨白藜芦醇对白血病K562细胞生长的影响及相关作用机制.方法 用不同浓度白藜芦醇作用于K562细胞,CCK-8法观察白藜芦醇对K562细胞生长增殖的影响,AnnexinV-FITC/PI双染法观察白藜芦醇对K562细胞凋亡的影响,PI染色法观察白藜芦醇对K562细胞周期的影响,Western blot法检测K562细胞中的Bcl-2、Bcl-xl、Bax、Cyclin D1蛋白.结果 白藜芦醇作用后的K562细胞的增殖被抑制并且凋亡增加,呈时间-剂量依赖性;同时,凋亡相关分子Bcl-2、Bcl-xl表达下调,Bax表达上调;白藜芦醇作用K562细胞后,G1期细胞增多,S期细胞减少,细胞周期调节蛋白Cyclin D1表达下降.结论 白藜芦醇可抑制K562细胞增殖并诱导其凋亡,其机制可能与下调细胞内Bcl-2、Bcl-xl、Cyclin D1表达并上调Bax的表达有关.     

藤黄酸调控白血病K562细胞GFI-1表达及对细胞增殖和凋亡的影响

目的: 探讨藤黄酸对白血病K562细胞增殖和凋亡的影响及可能的机制。方法：体外培养K562细胞,采用CCK-8法检测不同浓度藤黄酸(0、0.2、0.4、0.8、1.2、2.0μmol/L)处理24、48、72h后K562细胞增殖率;用流式细胞术检测K562细胞的凋亡和周期;采用实时荧光定量聚合酶链反应（RT-qPCR)）和Western blot法检测独立生长因子1(GFI-1) 的mRNA及蛋白表达水平。结果： 藤黄酸可以抑制K562细胞的增殖,并呈剂量、时间依赖性;0.4μmol/L藤黄酸处理24h后的K562细胞凋亡率增加;藤黄酸对GFI-1的mRNA表达无显著影响,藤黄酸可以下调GFI-1蛋白的表达;藤黄酸及蛋白酶体抑制剂（MG132）共处理后的K562细胞GFI-1蛋白水平较藤黄酸组升高;藤黄酸及氯喹（CQ）共处理后的K562细胞GFI-1蛋白水平与藤黄酸组无统计学差异（P>0.05）。结论： 藤黄酸可以有效抑制K562细胞的增殖并诱导细胞凋亡;藤黄酸可以诱导K562细胞G0/G1期及S期阻滞;藤黄酸通过蛋白酶体途径促进GFI-1蛋白降解。     

普鲁卡因通过miR-15a-5p/PI3K-AKT3途径调控乳腺癌细胞增殖、迁移和侵袭的机制

目的探讨普鲁卡因对乳腺癌细胞MCF-7增殖、迁移和侵袭的影响及其作用机制。方法以不同浓度(0.5、1.0、2.5、5.0、10.0 mmol/L)普鲁卡因处理MCF-7细胞,噻唑蓝(MTT)法检测细胞增殖;5.0 mmol/L普鲁卡因处理MCF-7细胞48 h后,Transwell法检测迁移和侵袭,实时荧光定量-聚合酶链反应(qRT-PCR)检测miR-15a-5p的表达。转染miR-15a-5p mimic至MCF-7细胞,检测细胞增殖、迁移和侵袭。靶基因预测软件预测miR-15a-5p和AKT3靶向结合位点,双荧光素酶报告基因实验检测二者靶向关系。转染miR-15a-5p inhibitor至MCF-7细胞,并用5.0 mmol/L普鲁卡因处理48 h,检测细胞增殖、迁移和侵袭,Western印迹检测蛋白激酶B(AKT)3的表达。结果与Con组相比,普鲁卡因能明显抑制MCF-7细胞增殖、迁移和侵袭(P<0.001),促进细胞中miR-15a-5p的表达(P<0.01)。过表达miR-15a-5p可抑制MCF-7细胞增殖、迁移和侵袭(P<0.001,P<0.01,P<0.05)。双荧光素酶报告基因实验证实miR-15a-5p和AKT3的靶向结合有关。抑制miR-15a-5p表达减弱了普鲁卡因对MCF-7细胞增殖、迁移、侵袭及AKT3表达的抑制作用(P<0.05)。结论普鲁卡因能够抑制乳腺癌细胞MCF-7的增殖、迁移和侵袭能力,其机制可能与调控miR-15a-5p/磷脂酰肌激-3-激酶(PI3K)-AKT3途径有关。     

miR-29b通过PI3K/AKT信号通路抑制白血病细胞的增殖

目的探讨miR-29b对K562白血病细胞增殖与凋亡的影响及相关机制。方法利用Lipofectamine~(TM)2000脂质体将miR-29b mimics,miR-29b NC转入白血病细胞K562中,实时荧光定量PCR法检测miR-29b表达,MTT法检测细胞活力,EdU染色法检测细胞增殖情况,流式细胞术检测细胞周期,Western印迹检测细胞中B淋巴细胞瘤(Bcl)-2、Bcl-2相关X蛋白(Bax),髓样细胞白血病(MCL)-1,细胞周期蛋白依赖性激酶(CDK)4,CDK6蛋白表达及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)激活情况。结果与miR-29b NC组比较,miR-29b mimics组miR-29b表达上调(P<0.01),细胞活力及细胞增殖率降低(P<0.01),细胞周期阻滞在G1期(P<0.01),Bcl-2,MCL-1,CDK4,CDK6,PI3K及p-AKT表达量均明显下调(P<0.01),Bax表达量明显上调(P<0.01)。结论 miR-29b mimics能显著抑制白血病细胞K562增殖、诱导细胞凋亡,可能与阻断PI3K/AKT信号通路有关。     

miR-205对黑色素瘤A375细胞增殖、黏附和迁移的影响及机制探讨

目的探讨miR-205对人皮肤恶性黑色素瘤细胞系A375增殖、黏附和迁移的影响及其可能机制。方法将miR-205 mimics用阳离子脂质体LipofectamineTM2000转染A375细胞,CCK-8法测细胞增殖变化;细胞黏附实验和划痕实验检测细胞黏附和迁移能力变化。Western blot检测PTEN、总AKT和磷酸化AKT蛋白表达变化。结果上调miR-205表达后实验组的细胞增殖、黏附和迁移能力均比空白对照组和阴性对照组减低(P均<0.05);Western blot结果显示上调miR-205表达能增加PTEN蛋白表达,减低磷酸化AKT蛋白水平,但对非AKT蛋白表达无影响。结论 miR-205可能通过调控PTEN/AKT通路抑制黑色素瘤细胞的增殖、黏附和迁移能力。     

反义寡核苷酸封闭癌基因MDM2提高白血病细胞化疗敏感性的研究

目的通过MDM2反义寡核苷酸（MDM2-AS）阻滞白血病K562细胞MDM2的表达,研究非p53通路下MDM2抑制的化疗增敏作用。方法细胞分为对照组、AS组、顺铂组和联合组,以RT—PCR和Western blot检测MDM表达,流式细胞仪检测处理后各组细胞的凋亡率。结果AS组细胞MDM2表达明显受抑制（P〈0．05）,联合组K562细胞凋亡率明显增加（P〈0．01）。结论在p53缺失情况下,以MDM2-AS反义封闭抑制MDM2表达,可增加白血病细胞的化疗敏感性,为白血病的临床治疗提供参考。     

齐墩果酸对K562细胞bax、bcl-xL mRNA表达的影响

目的 探讨齐墩果酸(OA)对人慢性粒细胞白血病K562细胞bcl-xL和bax基因表达的影响.方法 采用光学显微镜观察OA对体外培养K562细胞的增殖抑制情况;并用RT-PCR法检测bcl-xL和bax mRNA的表达.结果 与空白组和DMSO对照组比较,80 μmol/L OA处理细胞后,细胞增殖受抑制;OA用药组bax mRNA含量增加,bcl-xL mRNA含量减少.结论 OA可以诱导K562细胞凋亡,其机制可能通过上调bax基因表达及下调bcl-xL基因表达有关.     

反义nm23-H1基因对白血病k562细胞株侵袭及增殖的影响

目的应用RNAi(RNA interference)技术,通过对慢性髓细胞性白血病k562细胞株nm23-H1基因表达的抑制,初步研究nm23-H1基因对k562细胞株侵袭和增殖的影响。方法构建真核表达载体nm23-H1的RNAi重组体pGCsinm23(以下简称pGCsinm23),转染k562细胞株(转染组);设转染对照质粒的K562细胞为对照组。采用RT-PCR和Western blot法分别检测两组转染前后nm23-H1 mRNA及其蛋白质表达;采用平板克隆试验、软琼脂克隆形成试验和侵袭小室穿透试验观察转染前后细胞生长、增生能力以及体外侵袭能力的差异。结果 pGCs-inm23构建成功。转染组电泳结果显示nm23-1 mRNA和蛋白表达水平明显下降;与对照组比较,细胞生长速度加快,克隆形成率明显增高,软琼脂中细胞形成的集落体积增大且形成数量明显增多;穿膜细胞数亦明显增多(P<0.05)。结论 pGCsinm23可有效抑制K562细胞中nm23-H1的表达;nm23-H1基因可抑制白血病细胞增殖且对其侵袭能力有抑制作用。     

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.     

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.