EGFP标记的人肺癌裸鼠原位移植模型的建立

背景与目的 小鼠活体分子成像模型可以连续实时监测活体肿瘤的变化.本研究拟通过外科原位移植法建立表达绿色荧光蛋白的肺癌裸鼠原位移植模型并探讨其肿瘤生物学特性,从而建立一个良好的肺癌动物实验研究平台.方法 利用逆转录病毒转染法将增强型绿色荧光蛋白基因导人人肺癌大细胞系NCI-H460,采用外科原位移植法建立肺癌原位移植模型.定期通过小动物活体荧光成像系统观察肿瘤生长,利用相关性检验分析荧光面积和肿瘤体积之间的相关关系,并观察原位移植术后裸鼠的生存期和肿瘤转移情况.结果 模型建立后1周通过皮瓣在荧光体视镜下可观察到肺部肿瘤的绿色荧光,成瘤率为100%.荷瘤裸小鼠平均生存期为34.2天.解剖裸鼠观察到肿瘤侵及对侧肺、纵隔及肺门淋巴结、胸膜和膈肌,转移率分别为87.596、75%、25%和12.5%.肿瘤体积和荧光面积具有相关性(r=0.873,P=0.001).结论 外科原位移植法建立的表达EGFP的裸鼠肺癌原位模型是肺癌临床前研究的理想的实验工具.应用小动物活体荧光成像系统能够定量客观评价肿瘤在动物体内的生长、侵袭和转移,该模型可应用于肺癌的基础研究和新药开发.     

利用荧光素酶标记的肿瘤细胞建立活体成像动物模型

目的建立稳定表达荧光素酶的人乳腺癌细胞株并构建适用于小动物活体成像系统观察的裸鼠皮下移植瘤模型。方法采用脂质体将携带荧光素酶基因的质粒转染到人乳腺癌细胞株MCF-7中,G418筛选出稳定表达荧光素酶的单克隆细胞株。扩增后接种于裸鼠,建立裸鼠皮下移植瘤模型,通过活体动物成像系统监测肿瘤的生长过程。结果获得了高水平稳定表达荧光素酶的乳腺癌单克隆细胞株,该单克隆细胞株与母细胞系MCF-7具有相似的生长特性。将稳定表达荧光素酶的克隆接种于裸鼠皮下可成瘤,小动物活体成像系统能准确监测肿瘤细胞在体内的生长过程。结论成功建立了稳定表达荧光素酶的乳腺癌单克隆细胞株。采用活体动物成像系统构建的裸鼠皮下移植瘤模型是拓展肿瘤体内生长、转移及治疗相关研究的理想模型。     

人胃癌组织块裸鼠原位移植/转移模型的建立

 目的　用肿瘤组织块原位移植 ,建立人胃癌裸小鼠原位移植 /转移模型。方法　以人胃低分化腺癌细胞系接种于裸小鼠皮下 ,形成稳定传代的皮下移植瘤 ,再取该肿瘤组织块原位移植于裸鼠胃壁 ,观察移植肿瘤的生长状况、移植成功率和自发转移的发生率。结果　原位移植成功率 (成瘤率 )为 1 0 0 %、局部淋巴结转移率 1 0 0 %、远处淋巴结转移率 90 %、肝转移发生率为 75%。荷瘤鼠的中位生存期为 1 4周 ,晚期出现消瘦和全身衰竭。结论　该裸小鼠原位移植 /转移模型的生物学行为与人胃癌自然生长和转移过程相似 ,可作为一种有价值的工具用于胃癌转移机理和抗转移实验治疗的研究。     

4种人肾细胞癌原位移植裸鼠模型建立方法的比较

背景与目的:肾细胞癌是最常见的肾脏恶性肿瘤,起病隐匿,恶性程度高.研究旨在建立成功率高、稳定的人肾细胞癌原位动物造模方法.方法:采用人肾细胞癌细胞(786-0、ACHN)进行细胞悬液原位注射法、皮下成瘤后引瘤原代细胞悬液原位注射法、皮下成瘤后引瘤组织块原位移植肾包膜下法及皮下成瘤后引瘤组织块原位移植肾周筋膜内法建立原位移植裸鼠模型,采用PET/CT、H-E染色、免疫组织化学染色及血清生化检测等手段观察成瘤率及肿瘤生长情况,评估4种方法的建模效果.结果:人肾细胞癌皮下移植瘤裸鼠模型,ACHN组成瘤率(90%)高于786-0组(30%);人肾细胞癌原位移植裸鼠模型,786-0原位注射组、ACHN原位注射组、ACHN皮下移植瘤引瘤后原代细胞原位注射组、ACHN组织块肾包膜下包埋组和ACHN组织块肾周筋膜内包埋组的成瘤率分别为33%、80%、90%、100%和20%,其中以ACHN皮下成瘤后引瘤组织块原位移植肾包膜下法成瘤率最高,影像学及病理组织学检查结果显示符合低分化肾细胞癌.结论:成功建立4种人肾细胞癌原位移植裸鼠模型,其中以ACHN皮下成瘤后引瘤组织块原位移植肾包膜下法成瘤效果最佳,为肾细胞癌发病机制及靶向治疗的进一步研究提供理想的动物模型.     

人肺癌裸小鼠模型活体成像的动态观察

目的:建立稳定表达绿色荧光蛋白的人肺癌细胞系,并探讨小动物活体荧光成像系统在肺癌皮下移植瘤模型中的应用.方法:用慢病毒转染的方法建立表达绿色荧光蛋白的人肺癌细胞系NCI-H460-GFP,接种至裸小鼠体内建立皮下移植瘤模型,通过小动物活体成像系统连续5周观察肿瘤在小鼠皮下的动态生长情况.结果:建立了转染率接近100%的人肺癌NCI-H460-GFP细胞系,在体外及裸小鼠体内均能够长期稳定表达绿色荧光蛋白.活体荧光成像观察发现,1～4周随着肿瘤体积逐渐增大,平均荧光光子数逐渐增加;5周时随着肿瘤出现明显坏死,平均荧光光子数呈现下降趋势.结论:稳定表达绿色荧光蛋白的NCI-H460-GFP细胞系及其动物模型可以为肺癌研究提供理想的实验材料,应用小动物活体成像系统能够客观定量评价肿瘤在动物体内的生长情况.     

人类胃癌裸鼠淋巴结转移模型的建立

 目的　研究胃癌淋巴结转移动物模型的建立方法。方法　人类胃癌低分化细胞系SGC-7901体外培养、传代并扩增后，收集细胞行皮下种植成瘤，鼠间传代至第6代，以皮下肿瘤组织块原位种植于裸鼠胃壁建立动物模型。种植后第9周处死裸鼠，观察原位种植瘤生长、淋巴结转移及其他脏器转移情况，测定荷瘤裸鼠血清癌胚抗原（CEA）值。结果　原位移植瘤种植成功率100 ％，胃周淋巴结转移率93.3 ％，移植瘤可发生局部浸润及远处脏器转移，荷瘤裸鼠CEA值明显高于正常裸鼠（P＜0.01）。结论　应用SGC-7901细胞系可成功建立胃癌的淋巴结转移动物模型。     

胃癌细胞和组织块原位移植模型的建立及评价

[目的]采用两种方法建立胃癌裸鼠原位移植模型，并对两种模型进行评价。[方法]将胃癌肿瘤细胞悬液、胃癌裸鼠皮下瘤块采用OB胶粘贴法种植于裸鼠胃壁，形成原位移植瘤，观察和比较两种方法所建立的模型肿瘤移植成功率和自发转移的发生率。常规HE染色，观察胃癌原位移植瘤、胃癌淋巴结转移和肝转移的病理切片。免疫组化法检测胃癌原位移植瘤的MVD和VEGF的表达。[结果]细胞悬液种植的原位胃癌成瘤率为60％，淋巴结转移率和肝转移发生率仅为55．6％和33．3％；皮下组织块移植OB胶粘贴法的原位成瘤率100％，淋巴结转移率和肝转移发生率为93.3％和80％。皮下瘤块的原位移植瘤中MVD值和VEGF的表达高于细胞悬液法鼠移植瘤中MVD和VEGF的表达。[结论]两种原位移植和转移模型均具有人胃癌自然生长过程的特点，以胃癌裸鼠皮下组织块OB胶粘贴法为优，且肿瘤血管新生的能力较强。     

人子宫内膜癌瘤组织块活体移植瘤动物模型的建立

目的:研究建立人子宫内膜癌的组织块活体移植动物模型方法。方法:将生长状态良好的Ishikawa细胞浓度调整至1.5×107/ml,接种于BALB/c nude裸鼠腋下皮下(Ⅰ组),Ⅱ组生理盐水对照,观察其生长情况,待成瘤5-10mm时建立成瘤模型,收集Ⅰ组瘤组织块行活体移植于Ⅲ组小鼠,留取组织标本行病理检查。结果:成功建立了人子宫内膜癌Ishikawa细胞的BABL/c裸鼠皮下移植瘤模型和活体移植瘤模型,形成的肿瘤具备人肿瘤生物学特点。结论:建立的瘤组织块活体二次传代移植人子宫内膜癌的BABL/c nude小鼠皮下移植瘤模型具有人子宫内膜癌特征且成瘤率更高,为大批建立子宫内膜癌单位模型研究子宫内膜癌的发病机制和药物治疗提供了实验动物模型。     

通过活体动物光学成像系统建立乳腺癌动物模型

目的: 通过活体动物光学成像系统建立稳定表达荧光素酶的乳腺癌裸鼠移植瘤模型.方法: 用脂质体包裹带有neo抗性基因及荧光素酶报告基因的真核表达质粒pGL4.17[luc2/neo],体外转染人乳腺癌细胞株MCF-7,将获得稳定表达荧光素酶的乳腺癌细胞株(MCF-7-luc)进行皮下接种BALB/c裸鼠及尾静脉接种SCID小鼠,建立BALB/c裸鼠皮下移植瘤模型和SCID小鼠尾静脉移植瘤模型,通过活体成像系统监测体内肿瘤的生长及转移情况.结果: 经过筛选得到1个能够用于建立裸鼠皮下移植瘤模型及SCID小鼠尾静脉移植瘤模型的稳定表达荧光素酶的乳腺癌细胞株,并成功建立了乳腺癌裸鼠皮下移植瘤模型及SCID小鼠尾静脉移植瘤模型.结论: 通过活体动物光学成像系统成功获得的稳定表达荧光素酶的乳腺癌皮下移植瘤和尾静脉移植瘤模型,为人乳腺癌的相关研究提供了一个直观、方便、灵敏和可靠的平台.     

裸小鼠人胃癌原位移植模型的浸润转移特征

目的：建立裸小鼠人胃癌原位移植模型并观察其浸润转移特征。方法：BALB／C-nu／nu裸小鼠12只，以SGC-7901人胃癌细胞株裸鼠皮下移植瘤为材料，通过手术将瘤组织块移植到裸小鼠胃壁，待动物自然灏临死亡时进行系统解剖。结果：原位移植瘤全都成功，且所有动物均出现局部浸润及淋巴结转移，肝脏转移率为66．7％，50“的荷瘤鼠发生腹水，肿瘤晚期与周围脏器发生粘连，荷瘤鼠死于全身衰竭，中位数生存期为18周。结论：裸小鼠人胃癌席位模型的浸润转移特性接近临床病人，是研究人胃癌浸润转移的理想手段。     

Dermatomycosis in Goats in India

Summary: In an extensive survey involving 2176 goats 1.56% of goats manifested clinical lesions of ringworm infection. Animals below the age of 6 months were affected most (4.20%). The incidence of infection was higher during the winter months. T. verrucosum, T. mentagrophytes and M. gypseum were isolated from the skin scrapings of 12, 4, and 2 goats respectively. It is suggested that the infected animals could be a source of disease to human population in which the zoophilic dermatophyte invasion is characterized by severe inflammatory lesions of the skin. The zoonotic importance of different dermatophytes is stressed.
Zusammenfassung: In einer ausgedehnten Untersuchung an 2176 Ziegen wurden bei 1,56% klinische Zeichen einer Hautmykose festgestellt. Tiere, die jühger als 6 Monate alt waren, zeigten mit 4,20% am häfigsten Krankheitssymptome. In den Wintermonaten war die Erkrankungshäufigkeit am gröBten. T. vermcosum, T. mentagrophytes and M. gypseum wurden jeweils von 12,4 bzw. 2 Ziegen isoliert. Die inflzierten Tiere köinnen eine Infektionsquelle für die menschliche Bevölkerung darstellen und dort Mykosen mit stark entzündlichen Veränderungen auslösen. Die Bedeutung verschiedener Dermatophyten als Erreger von Zoonosen wird hervorgehoben.     

Trends in zygomycosis in children

Zygomycosis, or mucormycosis, is associated with significant morbidity and mortality in both children and adults. Studies in adults have shown an increase in the incidence of zygomycosis, particularly among haemtopoietic stem cell transplant (HSCT) recipients and patients with haematologic malignancies. There is a paucity of data on the epidemiology of zygomycosis in children. We performed a retrospective analysis to describe trends in zygomycosis between 1 January 2003 and 31 December 2010. We used the Pediatric Health Information System (PHIS) database to identify paediatric patients who were diagnosed with zygomycosis during the study period. Administrative data on diagnoses, demographics, underlying conditions and clinical experiences were collected. Summary statistics were calculated and tests for trend were conducted. We identified 156 unique patients with zygomycosis. The prevalence of zygomycosis did not significantly increase over time (P=0.284). The most common underlying condition was malignancy (58%) and over half received intensive care. Voriconazole utilisation among all hospitalised children significantly increased during the period (P=0.010). Our study demonstrates that the incidence of zygomycosis is not significantly increasing. During the time period there was a significant increase in the use of voriconazole among children.     

Increased apoptosis in cancer cells in vitro and in vivo by ceramides in transferrin-modified liposomes

Lysosomes are a promising therapeutic target for induction apoptosis in cancer cells due to lysosomal membrane permeabilization (LMP) leading to leakage of hydrolytic enzymes, especially the cathepsins, into the cytoplasm. We hypothesized that with the modification of the ceramide-loaded liposomes with transferrin (Tf), we would achieve both tumor targeting and increased delivery of lysosome-destabilizing agents, such as ceramides to lysosomes, to initiate LMP-induced apoptosis. We prepared Tf-modified (TL) and plain (PL) liposomes and loaded with short (C6)- or long (C16) N-acyl chain ceramides. Uptake, intracellular localization of liposomes, stability of the lysosomal membrane and release of cathepsin D were investigated on Hela cells by fluorescence microscopy and flow cytometry. Apoptosis was evaluated by binding of fluorescently-labeled Annexin V. Antitumor and pro-apoptotic effects of C6Cer-loaded Tf-liposomes were demonstrated in vivo in an A2780-ovarian carcinoma xenograft mouse model. TL were internalized specifically via the TfR-dependent endocytic pathway and localized within the endosome-lysosomal compartment. Ceramide-loaded Tf-liposomes significantly increased apoptosis compared with ceramide-free and ceramide-loaded non-modified liposomes. The treatment of cancer cells with TL led to increased LMP and cytoplasmic relocation of the intralysosomal cathepsin D. A strong antitumor and pro-apoptotic effect of C6Cer-loaded TL was also demonstrated in vivo in an A2780-ovarian carcinoma xenograft mouse model. The lysosomal accumulation of ceramides delivered by Tf-liposomes initiates the permeabilization of the lysosomal membranes required for the release of lysosomal cathepsins into the cytoplasm and initiation of the cancer cell apoptosis both in vitro and in vivo.     

Ringworm in animals in a farm in Assiut

Bericht über das Auftreten von Hautmykosen bei Tieren auf der Farm der Landwirtschaftlichen Fakultät der Universität Assiut. Von 70 Kälbern waren 42 pilzkrank, davon 22 durch T. mentagrophytes, 20 durch T. verrucosum. Von 180 Kühen waren 8 pilzbefalien, sämtlich durch T. mentagrophytes. Von 2 kranken Bullen wurde T. verrucosum isoliert. Auch die 3 Pferde und 2 Maultiere der Farm waren pilzinfiziert; Erreger war in diesen Fällen T. equinum. Auch ein Kalb war von T. equinum befallen. Die gleiche Pilzart wurde ferner von 3 Tierpflegern isoliert, die Pilzherde am Hals und an den Armen aufwiesen.     

Socioeconomic factors in cancer in Sweden

It is well known that certain cancers have shown clustering in socioeconomic groups, but limited data are available on recent results and time trends in such clustering. We determined standardized incidence ratios (SIR) for cancer, adjusted for age, period, region, parity and age at first childbirth among men and women in 6 socioeconomic groups based on the Swedish Family-Cancer Database. Persons had to be identified with the same socioeconomic status in the census of years 1960 and 1970, or of years 1960, 1970 and 1980; the comparison group was all people according to the same censuses. Cancers were followed from years 1970 to 1998 or from 1980 to 1998. Both increased and decreased SIRs were found, and a consistent pattern emerged, although the overall SIRs for cancer did not differ much, the lowest being for farmers (0.85) and the highest for professional men (1.07) and women (1.11). At individual sites, manual workers were at risk of tobacco-, alcohol- and occupation- and human papilloma virus-related cancers and at a decreased risk at most other cancers. Manual workers and farmers showed an excess of stomach cancer; professionals had an excess of melanoma and squamous cell skin cancer. Male and female SIRs correlated highly for manual and blue-collar workers and for professionals. The overall population-attributable fraction for selected sites was 16.7% for men and 10.9% for women and it was highest, over 50%, for lung cancer in both genders.     

Survey of changes in dietary preferences in cancer patients in China

Objective The aim of this study was to investigate the changes in dietary preferences in cancer patients in China and to determine the need for encouraging the adherence to a sensible diet among such patients.Methods A total of 468 cancer patients were interviewed using a self-designed questionnaire focusing on changes in the intake of specific foods. Data were analyzed using SPSS 16.0. Results Most patients completely avoided roosters and carp(73.1%), condiments(51.9%), and meat of aquatic species(40.4%). All other types of the specific foods were completely avoided by different subpopulations of the patients.Conclusion In addition to focusing on disease treatment, medical professionals need to help cancer patients overcome barriers associated with the customs of avoiding specific foods encompassed by the term ”fawu” and provide them with dietary guidance in order to prevent negative nutritional effects.     

Advances in cancer epidemiology in Japan

Epidemiologists in Japan have been performing calculations to estimate nationwide cancer incidence rates as well as 5‐year survival rates using population‐based cancer registry data. There have been remarkable changes in cancer incidence and/or mortality in cancers of the lung, liver and stomach, which were thought to be attributed to the changing impact of exposure to cigarette smoking, chronic hepatitis C virus infection and Helicobacter pylori infection, respectively. In systematic reviews providing evidence in risk/protective factors for cancer sites using case–control and cohort studies of the Japanese population, there were associations between cancer sites (esophagus, stomach, colo‐rectum, liver, pancreas, lung and breast) and various lifestyle factors. In the past 10 years, a hospital‐based case–control study at Aichi Cancer Center provided valuable evidence of gene‐environment interaction on the development of cancer [i.e., the effects of aldehyde dehydrogenase‐2 (ALDH2) polymorphism and heavy alcohol drinking on esophageal cancer, ALDH2 polymorphism and smoking on lung cancer, methylenetetrahydrofolate reductase polymorphism and heavy alcohol drinking on pancreatic cancer]. The database with stored DNA was also used and identified seven loci containing significant but low‐penetrance polymorphisms associated with the development of breast cancer. These findings together with established risk factors are likely to be useful to predict personalized breast cancer risk in East Asian women. In 2005, the Japan Multi‐Institution Collaborative Cohort (J‐MICC) study was launched to elucidate gene‐environment interactions as well as to confirm preclinical diagnostic biomarkers of cancer. J‐MICC, which has recruited 92,000 healthy individuals by the end of 2012, will follow the individuals until 2025.     

Foreign bodies in ENT in a teaching hospital in Eastern India

Objective

To evaluate the nature, common sites, modes of presentation of various foreign bodies (FB) in Ear, Nose and Throat (ENT).

Materials and methods

Observational retrospective study carried out at Vivekananda Institute of Medical Sciences, Ramakrishna Mission Seva Pratishthan. The study period was between Jan 2006 to Jan 2007. The information obtained from the hospital record books.

Results

Four hundred and eighty-two patients presented in the Department of Otorhinolaryngology of Vivekananda Institute of Medical Sciences, in the study period with FB in their ENT. Out of 482 pts, the commonest location of FB was to be in throat with 302 pts (62%) followed by ear with 119 pts (25%) and nose 61 pts (13%). Amongst the FB in throat the commonest was fish bone and the commonest site being tonsils. Artificial denture accounted for a significant number of 13 (4.3%). External auditory canal was the commonest site of FB lodgment in ear found in 118 patients (99.16%). Nasal FB were found in 61 patients; more common in pediatric age group (98.36%).

Conclusion

From this study we have found that FB lodgement is a very common complaint with which patients come to otolaryngologist. The commonest site of FB lodgement is in the throat. Most of the FB could be removed in emergency room (ER) with or without Local Anesthesia.     

整合素在急性白血病中的研究进展

急性白血病是一种早期造血干/祖细咆的恶性克隆性疾病.微小残留病和耐药被认为是其复发和难治的根源.近来,越来越多的证据显示白血病细胞通过与骨髓微环境中的基质细胞或细胞外基质相互作用促进其存活,并增强其对常规化疗药物的耐药.整合素是介导细胞与细胞外基质（细胞与非细胞成分）粘连的最主要黏附分子,在细胞增殖、存活等生物学过程中发挥关键作用,但作用机制尚不完全清楚.现就目前揭示的整合素及肿瘤微环境在急性白血病发生、发展中的作用进行综述.