HTLV-1 Tax 蛋白对T细胞中HMGB1调控的影响

目的探讨人T淋巴细胞白血病1型病毒（human T-cell leukemia virus 1,HTLV-1）Tax蛋白对人高迁移率族蛋白1（ high mobility group box 1,HMGB1）基因转录调控的影响。方法提取TaxN和TaxP细胞总RNA和蛋白质,通过real-time PCR和Western blot分析HMGB1 mRNA和蛋白质的表达情况;利用脂质体介导方法,将6个含有不同长度HMGB1调控序列的pGL3-HMGB1-luc瞬时转染至TaxN 和 TaxP 细胞,观察 HMGB1基因在不同 T 细胞中的转录活性;pCMV-Tax 与 pGL3-HMGB1-luc瞬时共转染至Jurkat细胞,观察Tax蛋白对HMGB1基因的转录调控影响;染色体免疫共沉淀（ ChIP）找寻Tax蛋白影响HMGB1基因转录调控的区段。结果 TaxP细胞中HMGB1 mRNA和蛋白质表达水平高于TaxN细胞。 TaxN和TaxP细胞中HMGB1调控趋势基本相似,均表现出3号质粒（pHLuc3,含有-504～＋83 HMGB1区段）的相对荧光素酶活性（HMGB1/neo）最高,但是6号质粒（含有-1163～＋83 HMGB1区段）却表现出 TaxP 细胞 HMGB1的转录活性明显高于 TaxN 细胞。pCMV-Tax与pGL3-HMGB1-Luc报告基因共转染到Jurkat细胞也显示,6号质粒（pHLuc6）中Tax促进HMGB1基因的转录。 ChIP分析证实了Tax蛋白可能富集在HMGB1的-1163～-1043区段。结论-504～-383可能是HMGB1基因转录激活的关键启动子区,Tax蛋白可能富集在HMGB1的-1163～-1043区段促进HMGB1基因转录。     

黄热病病毒检测方法学的研究进展

本文对黄热病病毒检测方法进行了综述。黄热病病毒的检测技术对该病的快速诊断,流行病学调查以及接种人群血清抗体的有效监测意义重大。正确快速地检测出黄热病病毒对保障我国人民健康安全和经济发展均有重大意义。     

K粉相关性膀胱炎的临床特点及治疗方法分析

目的：对氯胺酮(K粉)相关性膀胱炎患者进行疾病特征和治疗方法分析。方法24例K粉相关性膀胱炎患者行临床分析,包括一般资料、K粉滥用情况及不良症状情况,治疗应用抗菌药、胆碱能受体阻滞剂、肾上腺素能受体阻滞剂和抗抑郁类用药,效果不佳者行膀胱灌注。结果在患病者中,男性、低龄、文化程度低、用药时间长、用药剂量大、用药频率高及鼻吸患者比例较高(<0.05)。治疗后患者尿频、尿急、尿痛、尿失禁、血尿等不良症状均有明显改善(<0.05)。结论 K粉相关性膀胱炎的发病与K粉吸食时间、频率、药量有关,尿频、尿急、尿失禁是主要临床不良表现,对症用药和膀胱灌洗有一定疗效。     

酸敏感离子通道在中枢化学感受器呼吸调节中的作用

 目的：探讨脑室内注射酸化人工脑脊液（artificial cerebrospinal fluid, aCSF）引起的呼吸效应及酸敏感离子通道（acid sensing ion channels, ASICs）在此过程中的作用。方法：健康成年SD大鼠30只,随机分为aCSF（pH 7.4）对照组、aCSF(pH 6.5)组、ASICs阻断剂阿米洛利（amiloride）对照组、amiloride+aCSF（pH 6.5）组、ASIC1a阻断剂psalmotoxin 1 (PcTx1)对照组及PcTx1+aCSF(pH 6.5)组。通过膈肌肌电记录脑室内注射酸化aCSF后呼吸的变化;通过脑室内先注射阿米洛利和PcTx1再注射酸化aCSF的方法,观察酸敏感离子通道在中枢化学感受器呼吸调节中的作用。结果：脑室内注射酸化aCSF后,呼吸较注射前明显兴奋（P<0.05）;脑室内注射阿米洛利能完全阻断脑室内注射酸化aCSF引起的呼吸兴奋;脑室内注射PcTx1能部分阻断脑室内注射酸化aCSF引起的呼吸兴奋。结论：ASICs是参与中枢化学感受器呼吸调节的关键离子通道,ASIC1a则发挥了部分作用。     

HCV LNAzyme对病毒5'非编码区基因调控的特异性抑制作用

目的 探讨针对丙肝病毒5 '非编码区内源性核糖体进入位点的锁核酸核酶(LNAzyme)对病毒基因复制与表达的特异性抑制作用.方法 设计合成能切割HCV-5'-NCR-IRES位点的DNAzyme、硫代DNAzyme和LNAzyme.实验设对照组和实验组.对照组包括空白对照组、脂质体对照组和无关LNAzyme对照组.实验组包括DNAzyme、硫代DNAzyme组和LNAzyme组.以阳离子脂质体介导转染hepG2.9706细胞,用荧光定量PCR和化学发光技术分别监测24、48和96 h细胞培养上清液中HCV RNA含量及荧光素酶基因表达;四甲基偶氮唑蓝(MTT)法监测细胞活性.结果 加入核酶后,LNAzyme对HCV RNA复制和荧光素酶基因表达的抑制作用最强(P＜0.05),平均抑制率分别为48.02％和53.05％,且随用药时间延长,抑制率呈增高趋势,96 h后,平均抑制率分别为81.21％和84.25％.LNAzyme对细胞活性无影响.结论 LNAzyme能特异性抑制丙型肝炎病毒5 '非编码区的基因调控,且优于硫代修饰的DNAzyme.     

Majority Vote and Other Problems when using Computational Tools

Computational tools are essential for most of our research. To use these tools, one needs to know how they work. Problems in application of computational methods to variation analysis can appear at several stages and affect, for example, the interpretation of results. Such cases are discussed along with suggestions how to avoid them. The applications include incomplete reporting of methods, especially about the use of prediction tools; method selection on unscientific grounds and without consulting independent method performance assessments; extending application area of methods outside their intended purpose; use of the same data several times for obtaining majority vote; and filtering of datasets so that variants of interest are excluded. All these issues can be avoided by discontinuing the use software tools as black boxes.     

Does e-pain plan improve management of sickle cell disease associated vaso-occlusive pain crisis? A mixed methods evaluation

PurposeThere is limited application and evaluation of health information systems in the management of vaso-occlusive pain crises in sickle cell disease (SCD) patients. This study evaluates the impact of digitization of paper-based individualized pain plans on process efficiency and care quality by examining both objective patient data and subjective clinician insights.MethodsRetrospective, before and after, mixed methods evaluation of digitization of paper documents in Children's Hospital of Pittsburgh of UPMC. Subjective perceptions are analyzed using surveys completed by 115 clinicians in emergency department (ED) and inpatient units (IP). Objective effects are evaluated using mixed models with data on 1089 ED visits collected via electronic chart review 28 months before and 22 months after the digitization.ResultsSurveys indicate that all clinicians perceived the digitization to improve the efficiency and quality of pain management. Physicians overwhelmingly preferred using the digitized plans, but only 44% of the nurses had the same response. Analysis of patient records indicates that adjusted time from analgesic order to administration was significantly reduced from 35.50 to 26.77 min (p < .05). However, time to first dose and some of the objective quality measures (time from administration to relief, relief rate, admission rate, and ED re-visit rate) were not significantly affected.DiscussionThe relatively simple intervention, high baseline performance, and limited accommodation of nurses’ perspectives may account for the marginal improvements in process efficiency and quality outcomes. Additional efforts, particularly improved communication between physicians and nurses, are needed to further enhance quality of pain management.ConclusionThis study highlights the important role of health information technology (HIT) on vaso-occlusive pain management for pediatric patients with sickle cell disease and the critical challenges in accommodating human factor considerations in implementing and evaluating HIT effects.