两种方法建立人胃癌裸鼠移植瘤模型的比较

目的采用两种不同方法建立人胃癌裸鼠皮下移植瘤模型。方法人胃癌细胞悬液直接注射法:将处于对数生长期的人胃癌细胞悬液接种于裸鼠皮下;胃癌裸鼠瘤转瘤法:将胃癌细胞悬液接种入裸鼠,待肿瘤生长直径在8 mm左右时,处死裸鼠并取其肿瘤组织边缘新鲜的组织,接种于裸鼠皮下,形成皮下移植瘤。观察并比较两种方法所建立的模型肿瘤移植成功率及瘤体生长速度,实验结束后MSP法检测移植瘤组织中Reprimo基因的甲基化水平并采用免疫组织化学法检测移植瘤的Ki-67和CD31分子表达。结果细胞悬液直接注射组的瘤体成瘤速度较瘤转瘤组慢(P<0.05);瘤转瘤组的Reprimo基因甲基化水平及Ki-67和CD31的表达均高于细胞悬液直接注射组(P<0.05)。结论胃癌裸鼠皮下瘤转瘤法的瘤体生长状况优于胃癌细胞悬液直接注射法。     

不同时机协同给药诱导裸鼠脑胶质瘤细胞凋亡的对照研究

目的对比观察不同时机给予持续低剂量抗肿瘤细胞增殖和抗血管增生药物协同治疗裸鼠U251MG脑胶质瘤抑制脑胶质瘤生长的情况。方法分为两实验组,Ⅰ组,将2.5μl细胞悬液接种至6周龄雄性裸鼠脑白质内,接种次日预防性给药,是口服吲哚美辛(4 mg/kg)和腹腔注射榄香烯(70 mg/kg)协同给药;Ⅱ组,将等量的细胞悬液接种至4周龄雄性裸鼠脑白质内,接种后第20天治疗性协同给药,每组2只裸鼠,共4只裸鼠。给药20 d和30 d时,断椎处死裸鼠,脑标本石蜡切片3μm厚,分别行HE、胶质纤维酸性蛋白(GFAP)、CD34、TUNEL和PDCD-5等染色。结果预防性给药20 d裸鼠脑内未见明显增殖瘤细胞,亦无大量凋亡的瘤细胞,30 d时大量瘤细胞发生凋亡,GFAP呈阴性表达;治疗性给药裸鼠脑内仍可见胶质瘤细胞,20 d部分细胞发生凋亡,30 d大量细胞发生凋亡。结论预防性持续低剂量抗肿瘤增殖和抗血管增生药物协同治疗裸鼠脑胶质瘤,可有效遏制肿瘤的生长;治疗性给药虽能起到遏制肿瘤生长的目的,但治疗效果欠佳。     

转移性人小细胞肺癌裸鼠模型的建立

目的建立转移性人小细胞肺癌(SCLC)的裸鼠模型,探讨其用于人SCLC的治疗研究的可行性。方法 RPMI 1640培养基培养的人SCLC细胞株NCI-H446用无血清培养液重悬成1×107个/mL,取0.1 mL细胞悬液接种于BALB/c-nu裸鼠胫骨结节骨髓腔内,构建裸鼠模型并观察腿部肿瘤生长情况及肺等器官的转移情况。免疫组化染色检测小细胞肺癌标志物CD56及神经上皮干细胞标志物netsin的表达情况。结果经胫骨结节骨髓腔注射的方法注射细胞后裸鼠腿部均形成原位瘤,肺部转移6/6只,肝、脾等脏器未见转移灶。肿瘤组织细胞CD56阳性率>75%,nestin阳性率为4%～12%。结论经胫骨结节骨髓腔注射的方法可以形成弥散性肺部肿瘤,其他各器官则未见肿瘤块,可以为人SCLC实验性治疗的研究提供一种高效的实验模型。     

三氧化二砷对人食管癌细胞裸鼠移植瘤的抑制

目的:观察三氧化二砷对人食管癌细胞株Eca-109细胞裸鼠体内移植瘤生长的影响。方法:实验于2005-07/2006-07在解放军第四五八医院肿瘤中心实验室完成。①实验材料及方法:将体外培养的人食管癌细胞系Eca-109接种于裸鼠皮下,建立移植瘤模型。将20只裸鼠按随机数字表法分为4组(n=5),即生理盐水对照组、1μmol/L三氧化二砷组、4μmol/L三氧化二砷组和8μmol/L三氧化二砷组。各组均于瘤块移植后48h取不同剂量的药物0.2mL腹腔注射,给药5周。②实验评估:检测各组实体肿瘤生长情况并行常规病理观察。结果:20只Eca-109细胞移植裸鼠全部进入结果分析,无脱失。①裸鼠移植瘤生长情况:8μmol/L三氧化二砷组裸鼠对Eca-109细胞移植瘤的抑制率明显高于1μmol/L,4μmol/L三氧化二砷组和生理盐水对照组(P<0.05),其他各用药组之间移植瘤体积差异无显著性意义(P>0.05)。移植后4周和5周时,8μmol/L三氧化二砷组裸鼠Eca-109细胞移植瘤的抑制率分别为68.5%和80.7%,抑制作用呈剂量依赖性。②移植瘤组织病理学变化:生理盐水对照组细胞呈多角形,核圆形,核浆比例大,核分裂多见,可见异常核分裂。用药组瘤细胞孤立,核分裂少见,可见核染色质固缩,核边聚,核碎裂等凋亡现象。结论:三氧化二砷对人食管癌裸鼠移植瘤具有明显的抑制作用,并且呈一定的剂量依赖性。     

应用MEC-1与HG3细胞株建立慢性淋巴细胞白血病BALB/c裸鼠皮下移植瘤的模型

目的:通过对免疫缺陷鼠(BALB/c)不同部位、不同密度接种人慢性淋巴细胞白血病(CLL)细胞,探索裸鼠皮下移植瘤模型建立的条件。方法:通过慢病毒系统建立稳定表达绿色荧光蛋白(GFP)的人CLL细胞(MEC-1-GFP与HG3-GFP细胞)。首先将MEC-1-GFP细胞(5×10~7/ml)分别接种于裸鼠的前肢腋下、腹部及后肢腋下,观察不同接种部位对CLL细胞成瘤率的影响;其次将同等密度的MEC-1-GFP、HG3-GFP细胞(5×10~7/ml)分别接种于裸鼠的前肢腋下部位,观察两种细胞的成瘤时间及成瘤率。再次以同样接种方法,将不同密度的MEC-1-GFP与HG3-GFP细胞(1×10~7/ml与5×10~7/ml)分别接种于裸鼠的前肢腋下,观察不同接种密度的细胞成瘤时间及成瘤率。观察5周后,通过眼球取血获得外周静脉血,经EDTA抗凝及溶血素处理后,用流式细胞术检测GFP阳性的CLL细胞;同时,将荷瘤鼠处死,剥离肿瘤组织并制备冰冻切片,进行组织病理学检查。结果:成功获得稳定表达标记蛋白GFP的MEC-1与HG3细胞株;接种后裸鼠的前肢腋下、腹部及后肢腋下都形成了皮下移植瘤,且前肢腋下更有利于移植瘤的形成;接种密度为5×10~7/ml的MEC-1-GFP和HG3-GFP细胞均可形成皮下移植瘤,且5周后的成瘤率均为80%;接种密度分别为1×10~7/ml与5×10~7/ml MEC-1-GFP与HG3-GFP细胞均可有效地形成皮下移植瘤。流式细胞术检测显示,2组荷瘤鼠外周血中均未见GFP阳性的MEC和HG3细胞,组织病理检查表明CLL细胞向腹腔转移。结论:利用CLL细胞株MEC-1-GFP和HG3-GFP建立了CLL裸鼠皮下移植瘤模型,为进一步研究CLL的发病机制提供了实验工具。     

抑制Smac基因表达对胰腺癌裸鼠移植瘤生长影响的研究

[目的]探讨Smac基因对人胰腺癌裸鼠移植瘤生长的抑制作用.[方法]于裸鼠腋下接种200 μL 107/mL的PANC-1细胞悬液,建立人胰腺癌裸鼠移植瘤模型,将其分为三组：注射PBS液（A组）、转染空载体（B组）、转染Smac基因（C组）,比较三组移植瘤的大小、移植瘤细胞的凋亡率,移植瘤中Smac蛋白表达和微血管密度（MVD）.[结果]与A、B组比较,C组裸鼠移植瘤体积、质量和MVD显著减小;移植瘤细胞的凋亡指数和Smac蛋白的表达率均明显增高,其差异均有统计学意义（P〈0.05）.[结论]脂质体转染Smac基因能抑制人胰腺癌裸鼠移植瘤的生长,可能与该基因诱导胰腺癌细胞凋亡和抑制肿瘤血管形成有关.     

超声评估人乳腺癌原位移植裸鼠模型的初步研究

目的超声评估并选择最佳的MDA-MB-231细胞系裸鼠人乳腺癌模型。方法 54只裸鼠随机分为2组,每组27只,分别采用乳腺区皮下细胞悬液注射法、组织悬液注射法制作小鼠乳腺癌原位移植模型,比较两组的成瘤时间、出瘤率、肿瘤表面坏死出现时间,并用超声观察肿瘤的部分生物学形态改变。结果组织悬液注射法操作简便经济,成瘤时间短,肿瘤大小不规则,但有坏死和溃疡;细胞悬液注射法成瘤时间长,肿瘤形态规则,血管丰富。结论超声可以作为一种检测动物成瘤模型的手段,操作直观、立体,可视化。两种原位种植建立的裸鼠乳腺癌模型各有特点适合不同的实验需求。     

神经母细胞瘤原位荷瘤与转移瘤裸鼠模型建立的实验研究

目的用自建的细胞系QDDQ建立起原位荷瘤与转移瘤裸鼠模型,探讨神经母细胞瘤(Neuroblastoma,NB)的转移特性。方法取NB新鲜瘤组织,利用酶消化法,建立起体外细胞系,以(3～7)×107/ml浓度的单细胞悬液0.5 ml接种于裸鼠,建立荷瘤鼠模型。结果 NB原位荷瘤与转移瘤裸鼠模型成功建立,实验鼠共30只,致瘤25只,致瘤率为83.3%。10只原位荷瘤,15只转移荷瘤。结论建立NB细胞动物模型,对于进一步研究其转移特性、分子生物学特征,提供了良好的平台。     

移植性卡波西肉瘤动物模型的建立

目的建立模拟卡波西肉瘤发展过程的移植性卡波西肉瘤的动物模型。方法将36只裸鼠随机分为A、B、C、D四个实验组,每组各9只。卡波西肉瘤细胞体外培养并计数,调节细胞悬液浓度为1×10~8/L、1×10~9/L、5×10~9/L、1×10~(10)/L,各取0.2 ml分别接种于四组实验组裸鼠背部皮下。观察各组裸鼠肿瘤长出时间,肿瘤体积及颜色变化,待瘤体长到36 d时处死裸鼠,取下肿瘤组织进行组织病理学检查及免疫组化CD31、CD34、AE1/AE3、S100、HHV-8。结果卡波西肉瘤细胞株皮下注射移植成瘤率为100%,肿瘤组织经免疫组化CD31、CD34染色阳性证实为血管来源肿瘤。4个实验组中裸鼠肿瘤均由斑点逐渐形成斑块、结节,各实验组小鼠瘤体体积随时间延长而逐渐增大,不同细胞浓度的实验组小鼠瘤体体积差异无统计学意义(P>0.05)。第36天不同细胞浓度的实验组小鼠瘤体重量差异无统计学意义(P>0.05)。结论成功建立了卡波西肉瘤细胞移植动物模型,且注射卡波西肉瘤细胞浓度越高,长出肿瘤的时间越短。     

内皮抑素对ACHN RCC中缺氧诱导因子和血管内皮生长因子表达的影响

目的将已构建的带有内皮抑素基因真核表达载体质粒导入裸鼠ACHN肾细胞癌（renalcellcarcinoma,RCC）细胞中,研究内皮抑素基因对缺氧诱导因子（hypoxia-induciblefactor,HIF）和血管内皮生长因子（VEGF）表达的影响。方法荷瘤裸鼠随机分为3组,每组8只。干预组每只裸鼠瘤内3点注射复合物100μl,内含30μl梭华-SofastTM和30μgpSecES质粒;对照组每只裸鼠瘤内3点注射复合物100μl,内含30μl梭华-SofastTM和30μgpcDNA3.1质粒;空白组每只裸鼠瘤内3点注射生理盐水100μl。各组每只裸鼠间隔3d注射1次,连续注射3次。接种后第34天处死裸鼠,取肿瘤组织按SP法进行免疫组化染色,检测HIF-2α和VEGF的表达。结果干预组的ACHNRCC的生长较对照组和空白组明显受到抑制,干预组的HIF-2α和VEGF的表达较对照组和空白组低。HIF-2α和VEGF之间呈正相关（r=0.462,P〈0.05）。结论内皮抑素基因导入可以抑制荷瘤裸鼠ACHNRCC的生长,而且HIF-2α和VEGF在ACHNRCC的表达减少。HIF和VEGF二者之间存在正相关性。     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.     

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.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Systematic Reviews Published in the January 2011 Issue of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.     

Modified micro-superior percutaneously-assisted total hip: early experiences & case reports

Because of the extensile nature and familiarity of the standard posterior-lateral approach to the hip, a family of "micro-posterior" approaches has been developed. This family includes the Percutaneously-Assisted Total Hip (PATH) approach, the Supercapsular (SuperCap) approach and a newer hybrid approach, the Supercapsular Percutaneously-Assisted Total Hip (SuperPATH) approach. Such approaches should ideally provide a continuum for the surgeon: from a "micro" (external rotator sparing) posterior approach, to a "mini" (external rotator sacrificing) posterior approach, to a standard posterior approach. This could keep a surgeon within his comfort zone during the learning curve of the procedure, while leaving options for complicated reconstructions for the more practiced micro-posterior surgeons. This paper details one author's experiences utilizing this combined approach, as well as permutations of this entire micro-posterior family of approaches as applied to more complex hip reconstructions.     

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.     

Structure and function of "metalloantibiotics"

Although most antibiotics do not need metal ions for their biological activities, there are a number of antibiotics that require metal ions to function properly, such as bleomycin (BLM), streptonigrin (SN), and bacitracin. The coordinated metal ions in these antibiotics play an important role in maintaining proper structure and/or function of these antibiotics. Removal of the metal ions from these antibiotics can cause changes in structure and/or function of these antibiotics. Similar to the case of "metalloproteins," these antibiotics are dubbed "metalloantibiotics" which are the title subjects of this review. Metalloantibiotics can interact with several different kinds of biomolecules, including DNA, RNA, proteins, receptors, and lipids, rendering their unique and specific bioactivities. In addition to the microbial-originated metalloantibiotics, many metalloantibiotic derivatives and metal complexes of synthetic ligands also show antibacterial, antiviral, and anti-neoplastic activities which are also briefly discussed to provide a broad sense of the term "metalloantibiotics."