槲皮素对人急性B淋巴细胞白血病NOD/SCID小鼠细胞周期及黏附分子表达的影响

目的:探讨槲皮素对人急性B淋巴细胞白血病(B-ALL)NOD/SCID小鼠的细胞周期及黏附分子的影响。方法:对48只NOD/SCID小鼠于尾静脉注射5×106 Nalm-6细胞,建立ALL NOD/SCID小鼠模型,模型15 d后将ALL NOD/SCID小鼠随机分为3组:对照组(腹腔注射生理盐水(0.2 ml/只),环磷酰胺组(腹腔注射环磷酰胺100μg/kg)及槲皮素组(腹腔注射槲皮素3 mg/kg),治疗21 d后用流式细胞术检测Nalm-6细胞在G1、G2、M和S周期细胞百分率;计数治疗前后全血B淋巴细胞、Nalm-6细胞、中性粒细胞百分率及白细胞总数;采用双抗体夹心法测定治疗前后细胞间粘附分子-1(ICAM-1)、血管细胞粘附分子-1(VCAM-1)和P-选择素(P-selectin)蛋白表达。结果:与治疗前比较,治疗后槲皮素组ICAM-1、VCAM-1和P-selectin蛋白表达降低。血象显示,外周血中性粒细胞明显升高,而B淋巴细胞、白细胞和Nalm-6细胞明显降低; G0/G1期细胞增殖比例减少,S期和G2-M增多(P 0.05)。结论:槲皮素可能通过抑制ICAM-1蛋白表达而降低细胞间的粘附性,并将Nalm-6细胞阻滞在S期和G2-M期,对急性淋巴细胞白血病有明显的抑制作用。     

尾静脉注射K562细胞建立慢性髓系白血病小鼠模型及其鉴定

本研究通过给NOD/SCID小鼠尾静脉注射K562细胞的方式,建立人慢性髓系白血病(CML)-NOD/SCID小鼠髓外浸润模型,并通过组织病理学检查及RT-PCR检测BCR-ABL融合基因等进行模型鉴定。将24只NOD/SCID小鼠经137Cs全身照射,剂量270 cGy,吸收剂量率80 cGy/min,之后随机分为实验Ⅰ组、实验Ⅱ组和对照组,每组8只。照射后24 h内,实验组小鼠经尾静脉注射K562细胞,实验Ⅰ组:5×106/只,实验Ⅱ组:1×107/只;对照组注射生理盐水0.2 ml/只。持续观察小鼠的一般情况和生存时间,应用组织病理学检查和RT-PCR方法检测白血病细胞髓外浸润表现。结果表明,注射4-8周后,2个实验组均发生白血病细胞髓外浸润,提示成功建立CML/NOD-SCID小鼠髓外浸润模型。结论:应用尾静脉注射K562细胞的方法可成功建立CML/NOD-SCID小鼠髓外浸润模型。通过组织病理学检查及RT-PCR检测BCR-ABL融合基因可对模型进行鉴定。     

应用SCID和NOD/SCID小鼠构建急性髓系白血病模型成瘤率的比较

目的:比较应用SCID和NOD/SCID小鼠构建急性髓系白血病模型成瘤率的不同。方法:分别给SCID和NOD/SCID小鼠腹腔注射HL-60细胞,观察小鼠的一般情况、应用流式细胞术检测骨髓细胞CD33阳性率,病理学检查鉴定动物模型。结果:应用SCID小鼠构建急性髓系白血病模型成瘤率为30%,而应用NOD/SCID小鼠构建急性髓细胞白血病模型成瘤率为100%。结论:接种HL-60的NOD/SCID小鼠相对于SCID小鼠成瘤率高,小鼠发病率稳定,更适宜应用于白血病机制研究。     

尾静脉注射THP-1细胞构建急性髓系白血病NOD/SCID小鼠模型及其鉴定

目的:应用THP-1细胞构建NOD/SCID小鼠白血病模型。方法:将18只3-4周龄的雌性NOD/SCID小鼠,随机分为对照、模型A和模型B 3组(每组6只)。接种前连续2 d每只小鼠给予腹腔注射环磷酰胺2 mg/(kg·d),预处理后于24 h内接种细胞。模型组分别经尾静脉接种对数生长期THP-1细胞悬液1×10^7/只(A组)、5×106/只(B组),对照组鼠尾静脉注射等剂量生理盐水。观察一般情况,预处理前、接种细胞后7、14、21和28 d及处死时进行血常规检测、外周血白细胞分类,濒死前处死取材,组织切片病理检查。结果:模型组小鼠分别于接种细胞d 7和d 10开始出现竖毛、萎靡少动等现象,与对照组比较,模型A和B 2组小鼠体重于接种细胞21 d后明显下降(P<0.01),建模28 d后模型组白细胞数明显升高,差异有统计学意义(P<0.01),其中以接种1×10^7/只的模型A组最为显著。病理组织切片结果提示,模型组小鼠脾脏均可见白血病细胞弥漫性浸润。免疫组织化学结果提示,白血病细胞抗人CD13结果阳性,证实模型建立成功。结论:NOD/SCID小鼠经腹腔注射CTX预处理后,每只小鼠尾静脉注射THP-1细胞1×10^7或5×10^6个均可成功构建急性髓系白血病动物模型,符合急性髓系白血病的生物学特点,高浓度成瘤更快。     

应用含标记基因K562细胞制备的小鼠白血病模型

背景:文献报道应用NOD/SCID及SCID小鼠可以建立移植性人白血病小鼠模型,但由于小鼠存在免疫缺陷,使得在自体干细胞移植及移植后相关过继免疫治疗方面的研究受到限制和影响.目的:探索用SPF级Balb/c小鼠和转染GFP及NeoR基因的K562细胞株制各自血病模型的方法.方法:实验分为5组,A、B组和C、D组分别经x射线照射2Gy和3Gy,24 h后取对数生长期的K562(GFP+/Neo+)细胞,A、C组尾静脉注射2×106个/只;B、D组尾静脉注射5×106个/只:E组为正常对照组.观察小鼠生存时间,进行骨髓细胞及外周血白细胞分类,采用流式细胞仪测定GFP阳性细胞及PCR方法测定Neo基因.结果与结论:实验各组在接种5-7 d时发病.分别于30,23,24,17 d内全部死亡;生存天均显著短于正常对照组(P<0.01).体质量均较正常对照组显著下降(P<0.05),小鼠的向血病发病率为100%,无自发缓解.随接种细胞数量的增加和照射剂量的增加,小鼠的存活时间缩短,且外周血及骨髓中白血病细胞所占比例增加,流式细胞仪测定及PCR方法也证实了GFP+细胞和NeoR基因在肝、脾中的存在.结果证实Balb/c小鼠经照射后从尾静脉注射K562细胞株可以制备抉得白血病小鼠模型.     

K562/NOD-SCID小鼠白血病模型的建立

本研究探讨人CML急性变的白血病细胞在NOD—SCID小鼠体内建立白血病模型的方法并研究其生长特性。首先将K562细胞接种于全身受照射后的裸鼠，待皮下成瘤后取出局部瘤块，选取无坏死的瘤组织制成瘤细胞悬液，再腹腔接种于全身受照射后的NOD—SCID小鼠。结果表明：成功建立全身播散的白血病模型。4周时外周血涂片可见白血病细胞，晚期浸润肝、脾、骨髓等造血器官，白细胞上升到接种前的8—10倍，血涂片中白血病细胞达20％-30％。腹腔局部出现瘤块，多位于腹腔内或大网膜，较少累及其他器官。结论：腹腔接种K562瘤细胞于全身照射后的NOD—SCID小鼠能建成全身播散的白血病模型，该模型较好地反映白血病在体内的演变过程，是进行新药疗效试验、生物导向治疗及基因治疗的理想工具。     

人慢性粒细胞白血病模型鼠的建立

通过SCID小鼠与NOD/Lt品系(T淋巴细胞、B淋巴细胞、自然杀伤细胞缺陷,循环补体缺乏,抗原呈递细胞分化及功能不良)回交得到的NOD/Ltsz-SCID/SCID小鼠(简称NOD/SCID小鼠).目前,NOD/SCID小鼠白血病模型的建立多采用经尾静脉接种较大数量的白血病细胞[1-3],虽然可以保证成功植入,但对研究小鼠的机体免疫反应实验十分不利,主要原因在于植入的肿瘤细胞不再分裂繁殖且小鼠本身为免疫缺陷鼠.     

建立人急性粒细胞白血病M_2裸鼠模型

背景:人实体瘤细胞容易在小鼠体内成瘤,但人荷瘤白血病模型很难构建.通过放射线或环磷酰胺抑制裸鼠免疫系统预处理,可构建低成本、高稳定性的裸鼠模型.目的:探讨融合基因AML/ETO阳性的人急性粒细胞白血病M2白血病细胞Kasumi-1在BALB/c裸鼠体内建立白血病模型的方法.方法:将BALB/c裸鼠以抽签法随机分3组:环磷酰胺组腹腔注射环磷酰胺连续2 d后,尾静脉注射Kasumi-1细胞8×10~5/只;照射组给予X射线全身照射,照射当天尾静脉注射Kasumi-1细胞;无预处理组未作任何处理,尾静脉注射Kasumi-1白血病细胞.另取3只正常BALB/c裸鼠作为正常对照.检测外周血涂片、骨髓涂片,流式细胞仪检测骨髓细胞免疫分型,RT- PCR 检测白血病细胞瘤负荷,FISH检测骨髓细胞AML/ETO融合基因阳性细胞百分比.结果与结论:未经任何预处理裸鼠建模14 d血涂片中白血病细胞达3.5%,骨髓中肿瘤细胞百分比可达40%以上,与FISH和流式细胞仪检测白血病细胞比例一致,且随着接种时间延长,瘤负荷不断增加.全身照射和环磷酰胺注射后的裸鼠瘤负荷高于无预处理组,但仍可带瘤生存60 d.正常裸鼠外周血单个核细胞RT-PCR未发现有融和基因AML/ETO,其他3组均可见融和基因AML/ETO的mRNA表达.提示给予环磷酰胺组和X线照射预处理或单纯尾静脉接种Kasimi-1细胞均可建立急性粒细胞白血病M2裸鼠模型.     

雷公藤红素对人急性早幼粒白血病荷瘤模型小鼠黏附分子及细胞周期的影响

目的:观察雷公藤红素对人急性早幼粒白血病(APL)荷瘤模型小鼠黏附分子及细胞生物学特性的影响.方法:将18只SCID beige小鼠尾静脉注射NB4细胞株(5x106/只)以构建人APL荷瘤模型,然后随机分为荷瘤组、三氧化二砷组和雷公藤红素组,另取6只不造模并设为对照组;3周后向对照组和荷瘤组腹腔注射生理盐水并进行对照...     

多药耐药基因转染的脐血细胞对白血病小鼠化疗的骨髓保护作用

目的探讨转染多药耐药(mdr1)基因的脐血单个核细胞(MNC)对急性髓系白血病小鼠的骨髓保护作用及疗效。方法通过逆转录病毒介导的方法将含有人全长cDNAmdr1基因导入脐血MNC,即逆转录病毒上清液与脐血MNC体外共培养;将接种人髓系白血病细胞系(HL60细胞)的SCID小鼠分成3组A组(观察组)经鼠尾静脉注射转染mdr1的脐血MNC2×106/只,共2次;B、C两对照组小鼠以同样剂量、方法分别注射未转染mdr1的脐血MNC和等容积的生理盐水。3组白血病SCID小鼠在每周递增高三尖杉酯碱剂量化疗下,通过检测小鼠外周血白细胞数、瘤细胞阳性率、组织病理和HL60细胞表面抗原(CD33)等观察转基因小鼠和对照小鼠对抗癌药物的耐受性及抗肿瘤疗效。同时分别采用PCR技术、免疫组化方法和柔红霉素排出试验检测mdr1基因在小鼠体内的表达和功能。结果①体外成功地将mdr1基因导入脐血MNC,转染率达30%左右;②用HL60细胞2×106接种于经亚致死量照射的SCID小鼠可成功制成白血病动物模型;③采用程序性移植转基因细胞方法成功建立了mdr1转基因脐血细胞移植小鼠模型,并可作为白血病临床前期体内评价mdr1基因保护骨髓作用;④转基因脐血细胞移植小鼠对高三尖杉酯碱耐受性高于正常剂量的5~6倍,外周血白细胞维持在3.0×109/L左右,外周血涂片瘤细胞降至5%以     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.