多光谱小动物活体成像技术在骨性关节炎模型评价中的应用

目的：利用多光谱小动物活体成像技术动态观察和评价大鼠骨性关节炎模型的应用价值。方法：取3月龄SD雄性大鼠15只（180±20）g,利用碘乙酸关节腔注射法建立骨性关节炎模型：左膝关节腔注射碘乙酸50μl建立OA模型组,右膝关节腔注射等量无菌生理盐水做为正常对照。分别于建模2、4周时进行X-ray活体成像观察,测算骨密度。4周后处死大鼠,取双侧关节进行组织病理学观察,X线和组织病理学参考Kellgren-Lawrence和Collins进行分级。结果：OA模型造模成功,活体成像观察发现与对照组相比,模型组出现关节面不平整、骨赘形成、关节变形、软骨缺损等典型骨性关节炎表型,并伴有股骨远端骨密度下降明显（P〈0.01）,胫骨近端骨密度下降不明显（P〉0.05）,病理学结果一致;2周后模型组膝关节接近Collins 1级、Kellgren-Lawrence 2级,4周后模型组膝关节接近Collins 4级、Kellgren-Lawrence 3级;对照组关节面平整,关节间隙正常,软骨未见明显缺损,Collins及KellgrenLawrence 0级,股骨远段及胫骨近端骨密度正常。结论：多光谱小动物成像技术可用于骨性关节炎模型的影像学活体动态观察及骨密度检测,对骨性关节炎模型评价和相关研究具有重要意义。     

MR扩散峰度成像对非酒精性脂肪性肝病诊断价值

目的探讨扩散峰度成像(DKI)对非酒精性脂肪性肝病(NAFLD)定量分级诊断及早期检出非酒精性脂肪性肝炎(NASH)的价值。方法采用高脂饮食喂养新西兰大白兔,构建不同等级非酒精性脂肪性肝病动物模型。采用3.0 T磁共振、关节表面柔线圈,进行肝脏扩散峰度成像(b=0、400、800 s/mm2)和磁共振扩散加权成像(DWI),分别获取MD值、MK值和ADC值,对所有肝脏标本进行非酒精性脂肪性肝病活动度评分。采用Spearman相关性分析评估不同等级与磁共振参数的相关性。采用ROC明确各磁共振参数诊断不同等级非酒精性脂肪性肝病的效能。采用比较ROC分析不同模型参数在早期诊断非酒精性脂肪性肝炎中的价值。结果ADC值和MD值与非酒精性脂肪性肝病等级程度呈负相关(ρ=-0.52,-0.904;P<0.05)。MK值与疾病等级程度呈正相关(ρ=0.761;P<0.05)。MD值和MK值在临界组和NASH组间差异有统计学意义(MD:1.729±0.114比1.458±0.240μm^2/ms;MK:1.096±0.079比1.237±0.180;P<0.05)。MD值和MK值诊断NASH的ROC曲线下面积分别为0.955和0.905,且均优于ADC值(AUC=0.727)(P<0.05)。结论扩散峰度成像可对非酒精性脂肪性肝病进行分级诊断,且早期诊断非酒精性脂肪性肝炎的价值优于传统磁共振扩散加权成像。     

人羊水干细胞移植对大鼠四氯化碳诱导性肝硬化的治疗作用及其机制

【目的】探讨人羊水干细胞移植对大鼠四氯化碳诱导性肝硬化的治疗作用及其机制。【方法】采用60%的四氣化碳植物油皮下注射7周制造肝硬化大鼠模型,再随机分成对照组(注射等体积PBS)、hAFSC移植组和hAFSC移植后基质细胞衍生因子Ⅰ(SDF-1)受体CXCR4的特异性阻断剂普乐沙福(AMD3100)干预组,处理3周后处死所有大鼠,检测大鼠血清碱性磷酸酶(ALP)、白蛋白(ALB)和胆固醇(Ch),按Ishak评分标准评估肝硬化情况。体外实验分为3个组:对照组(等剂量PBS)、AMD3100组、AMD3100+SDF-1组,观察不同处理因素对hAFSC的存活、迁移、黏附的影响。【结果】与对照组比较,移植组的血清ALB和Ch均显著升高(P<0.05),ALP无显著变化(P>0.05),而干预组的ALP、ALB及Ch与对照组比较差异均无统计学意义(P>0.05)。移植组与对照组肝组织纤维化评分比较差异有统计学意义(P=0.005),移植组与干预组也有显著差异(P=0.022)。体外实验结果表明:SDF-1能缓解AMD3100对hAFSC的迁移、存活、黏附的抑制(P<0.05)。【结论】hAFSC经静脉肝内移植可减轻大鼠四氯化碳诱导性肝硬化病变程度,其机制与SDF-1/CXCR4调控有关。     

National prescribing practices of adolescent psychiatrists for psychotropic medications in outpatient care in Finland

A possible link between experiences of dysfunctional parental rearing and late behavioural outcomes could consist of the development of dysfunctional assumptions about self and others in relationship. This hypothesis has previously been addressed in psychiatric patients. Accordingly, the confounding influence of mental status on the endorsement of dysfunctional assumptions cannot be wholly excluded. The present study, in which a community sample of 233 subjects was enrolled, was planned to contribute to clarifying this issue. Subjects completed the EMBU to assess perceptions of parental rearing and two measures of dysfunctional assumptions, or dysfunctional working models: the DAS and the DWM-S. It was predicted that experiences of caring parental rearing would be negatively correlated with scores on both the DAS and the DWM-S. Conversely, it was predicted that experiences of dysfunctional parenting would be positively correlated with scores on the two scales. Both predictions were supported by the results. Perceptions of dysfunctional maternal rearing contribute most to high scores both on the DAS and the DWM-S.     

慢病毒介导shRNA沉默Nicastrin基因的人永生化角质形成细胞模型构建

目的 构建Nicastrin(NCT)基因的RNA干扰慢病毒表达载体,并在人永生化角质形成细胞(HaCaT细胞)上鉴定其沉默效率,构建NCT基因稳定下调的人永生化角质形成细胞模型,为后续研究NCT基因下调对角质形成细胞的生物学行为影响奠定实验基础.方法 设计3个靶向NCT基因特异性短发卡RNA(shRNA)表达序列并连接到慢病毒表达质粒pGLV3/H 1/GFP+ Puro中,成功构建慢病毒重组表达质粒,并通过测序鉴定.将构建的重组表达质粒与包装质粒共转染293T细胞,产生慢病毒颗粒,并测其滴度.将HaCaT细胞分为空白组(未感染病毒)、阴性对照组(NC组,感染空载病毒LV3-shNC)和干扰组(感染NCT-shRNA1、2、3慢病毒).流式细胞仪检测慢病毒转染效率,实时荧光定量PCR(qRT-PCR)、Western印迹检测靶基因的沉默效率,筛选出沉默效果最佳的干扰序列.结果 测序表明,NCT-shRNA慢病毒重组表达质粒构建成功.重组表达质粒与包装质粒共转染293T细胞后产生的慢病毒颗粒滴度为109 TU/ml.慢病毒感染HaCaT细胞后,流式细胞仪检测,慢病毒转染效率大于95％.qRT-PCR结果,与NC组相比,干扰组NCT mRNA表达量明显下降,其中NCT-shRNA1组NCT基因沉默效果最佳,干扰效率达到75％.Western印迹结果,与NC组相比,shRNA1组NCT蛋白表达抑制率为71.7％.结论 成功筛选出高效NCT-shRNA干扰序列,构建NCT-shRNA慢病毒重组表达载体,并构建NCT基因稳定下调的人永生化角质形成细胞模型.     

Identity by descent estimation with dense genome‐wide genotype data

We present a novel method, IBDLD, for estimating the probability of identity by descent (IBD) for a pair of related individuals at a locus, given dense genotype data and a pedigree of arbitrary size and complexity. IBDLD overcomes the challenges of exact multipoint estimation of IBD in pedigrees of potentially large size and eliminates the difficulty of accommodating the background linkage disequilibrium (LD) that is present in high‐density genotype data. We show that IBDLD is much more accurate at estimating the true IBD sharing than methods that remove LD by pruning SNPs and is highly robust to pedigree errors or other forms of misspecified relationships. The method is fast and can be used to estimate the probability for each possible IBD sharing state at every SNP from a high‐density genotyping array for hundreds of thousands of pairs of individuals. We use it to estimate point‐wise and genomewide IBD sharing between 185,745 pairs of subjects all of whom are related through a single, large and complex 13‐generation pedigree and genotyped with the Affymetrix 500 k chip. We find that we are able to identify the true pedigree relationship for individuals who were misidentified in the collected data and estimate empirical kinship coefficients that can be used in follow‐up QTL mapping studies. IBDLD is implemented as an open source software package and is freely available. Genet. Epidemiol. 2011. © 2011 Wiley‐Liss, Inc. 35: 557‐567, 2011     

我校护理专科生实践教学体系的构建与实施

目的建立新的实践教学体系，旨在提高学生的职业能力，使培养的人才更加符合社会及职业岗位需求。方法自2005年始，采用新的教学理念和教学方法设计培养方案，使实践教学体系贯穿整个培养过程。学生临床实习结束后，我们分别到河南省南阳市的几家医院，对该年级学生的实习情况作了跟踪调查。结果93．0％的临床带教老师认为．该届学生和往届学生相比整体素质较高、工作能力较强。结论新的实践教学体系突出了教学过程的实践性、开放性和职业性，使培养的人才更加符合社会需求。     

Unique Hippocampal Changes and Allodynia in a Model of Chronic Stress

Sustained stress can have numerous pathologic effects. There have been several animal models for chronic stress. We tried to identify the changes of pain threshold and hippocampus in a model of chronic stress. Male Sprague-Dawley rats were kept in a cage filled with 23℃ water to a height of 2.2 cm for 7 days. Nociceptive thresholds, expressed in grams, were measured with a Dynamic Plantar Aesthesiometer. Golgi staining was used to identify hippocampal changes. To demonstrate how long allodynia was lasting, behavioral test was repeated daily on another experiment. Compared to control group, chronic stress group showed bilateral mechanical hyper-responsiveness on days 5 (P = 0.047) and 7 (P = 0.032). In general, dendrite atrophic changes within hippocampus of chronic stress model were much more prominent in comparison with control. Compared to control, decreased spine number (P < 0.001) and spine length (P < 0.001) on Golgi staining were seen in the hippocampus of animals with chronic stress. Bilateral mechanical hyperresponsiveness was recovered on day 19 in animals with chronic stress. Chronic stress may bring about central sensitization and hippocampal changes in rats.     

靶向CT对比剂在结核急性感染动物模型中的初步观察

目的 探讨抗85B和ESAT-6鼠单克隆抗体靶向CT对比剂对小鼠急性感染肺结核动物模型显示的可行性.方法 采用已纯化的抗85B和ESAT-6鼠单克隆抗体与碘原子耦联制备靶向CT对比剂,计算对比剂溶液中抗体含量和结合127Ⅰ含量,稀释后配制成5μg I/ml备用.将20只小鼠急性感染肺结核动物模型通过随机数字表法随机分为4组,每组5只并编号,分别记为靶向对比剂85B组和ESAT-6组、普通对比剂组和对照组.普通对比剂组与85B组注射含碘浓度相同的碘海醇稀释液；对照组注射pH 7.4的磷酸盐缓冲液(PBS).分别于注射对比剂前、后不同时间(即刻,6、12、24 h)行微CT(Micro-CT)扫描,观察小鼠肺内病变的显示情况,判断指标为病变CT值,绘制CT强化-时间曲线,并计算强化率.两样本均数比较用t检验,多个样本均数比较用方差分析.结果 实验所制备的2.52 ml抗85B对比剂溶液中含有210～ 255μg抗体、10.5 ～ 16.6μg 127Ⅰ;2.93 ml抗ESAT-6对比剂溶液中含有抗体147μg、结合127Ⅰ约20.58 μg.靶向对比剂组小鼠肺内病变的CT值随时间延长而逐渐升高,在注射对比剂后12 h(t12)病变强化明显,在注射对比剂24 h(t24)后肺内病变仍有强化,且与普通对比剂组小鼠肺内病变的CT值相比[85B组t12=(-125.04±13.58)HU,t24=(-117.37±12.28)HU和ESAT-6组t12=(-122.14±19.01)HU,t24=(-114.23±17.08)HU],差异均有统计学意义(2组24 ht值分别为4.05和6.39,P值均＜0.05).对照组小鼠肺内病变在注射PBS前后,均未见强化.结论 制备的抗85B和ESAT-6结核特异抗体靶向CT对比剂在小鼠肺结核动物模型中具有一定靶向性,为结核相关靶向对比剂的研究奠定了实验基础.     

通气侧肺吸入伊洛前列素对肺动脉压和缺氧性肺血管收缩的影响

目的 探讨雾化吸入伊洛前列素对大鼠单肺通气(one-lung ventilation,OLV)期间肺动脉压和缺氧性肺血管收缩(hypoxic pulmonary vasoconstriction,HPV)的影响。方法 雄性SD大鼠30只,随机分为A、B、C三组,每组10只。原位离体肺灌注模型建立后,调整气管导管深度致左侧肺OLV,FiO_2为100%,打开连接于呼吸回路的雾化器,A组吸入生理盐水,B组吸入伊洛前列素0.05μg·kg-1·min-1,C组吸入伊洛前列素0.1μg·kg~(-1)·min~(-1),记录灌注10min(T_1)、雾化吸入10min(T_2)和OLV 1h(T_3)的平均肺动脉压(MPAP)及左心房引流液氧分压(PaO_2)。通过测量T_1、T_2和T_3时的左心房引流液PaO_2计算氧合指数(oxygenation index,OI)。实验结束分别取双侧肺组织进行电镜检查。结果 与T_1时比较,T_2、T_3时三组MPAP明显升高(P0.05);T_2、T_3时B、C组MPAP明显低于A组(P0.05),且C组MPAP明显低于B组(P0.05)。与T_1时比较,T_2、T_3时三组OI明显降低(P0.05);T_2、T_3时C组OI明显高于B组(P0.05)。相对于B、C组通气侧,A组通气侧与A、B、C组非通气侧Ⅱ型肺泡上皮细胞核膜外突内陷,部分板层小体排空。结论 在大鼠原位肺灌注模型中,单肺通气期间雾化吸入伊洛前列素能明显降低平均肺动脉压,减少肺内分流并增加氧合。