三磷酸腺苷生物荧光液闪测定法检测活细胞计数

为建立一种检测活细胞数的方法,作者采用三磷酸腺苷(ATP)生物荧光液闪测定法检测小鼠纤维瘤细胞株L929.ATP标准曲线工作范围为10 -9～10-5mol/ml,相关系数r=0.9963(P＜0.001);当活细胞数在3×1 02～106个/ml时与发光计数值有良好的线性关系,r=0.9922(P＜0.001),变异系数 CV = 1%～3%.用ATP生物荧光液闪测定法检测活细胞数灵敏、简便、准确、重复性好,用液闪计数仪即可完成测定,有较大的实用价值.     

ATP生物荧光肿瘤药敏检测技术在乳腺癌化疗中的应用价值

目的　探讨ATP生物荧光肿瘤药敏检测技术 (ATP TCA )在乳腺癌化疗中的临床意义及应用价值。方法　选用 10种常用乳腺癌化疗药物 ,应用ATP TCA对 40例乳腺癌改良根治术标本、淋巴结活检和胸腔积液标本进行药敏检测 ,评估肿瘤细胞对化疗药物的敏感性。结果　ATP TCA对乳腺癌标本的可评价率为 90 .0 % ,化疗药物对乳腺癌的杀伤作用具有较强的个体差异性 ,10种化疗药物的敏感性分别为 :氟尿嘧啶 ( 5 Fu) 3 3 .3 %、顺铂 (DDP) 3 7.5 %、环磷酰胺 (CTX ) 2 9.2 %、足叶已甙 (VP 16)16.7%、丝裂霉素 (MMC) 2 2 .0 %、表阿霉素 (EPI) 41.7%、诺维本 (NVB) 45 .8%、阿霉素 (ADM ) 41.7%、泰素 (PTX) 5 4.2 %、羟基喜树碱 (HCPT) 2 5 .0 %。初步研究表明ATP TCA体外检测结果与实际临床疗效具有良好的相关性。结论　ATP TCA是准确的、可靠的肿瘤药敏检测技术 ,其检测结果与临床实际疗效具有良好相关性 ,可用于指导乳腺癌术后化疗     

SM5-1-Conjugated PLA nanoparticles loaded with 5-fluorouracil for targeted hepatocellular carcinoma imaging and therapy

SM5-1 is a humanized mouse antibody which has a high binding specificity for a membrane protein of about 230 kDa overexpressed in hepatocellular carcinoma (HCC), melanoma and breast cancer. In this study, SM5-1-conjugated poly d, l (lactide-coglycolide) (PLA) PLA containing Cy7 (PLA-Cy7-SM5-1) was prepared to study the targeting specificity of the bioconjugate to HCC-LM3-fLuc cell. Then, SM5-1-conjugated PLA containing 5-fluorouracil (5-FU) (PLA-5FU-SM5-1) and PLA containing 5-FU (PLA-5FU) were prepared for treatment of subcutaneous HCC-LM3-fLuc tumor mice. The results showed that PLA-5FU-SM5-1, PLA-5FU and 5-FU induced a 45.07%, 23.56% and 19.05% tumor growth inhibition rate, respectively, on day 31 post-treatment as determined by bioluminescent intensity. In addition, in order to evaluate the antitumor efficacy of PLA-5FU-SM5-1, HCC-LM3-fLuc cells were injected into the liver to establish the experimental orthotopic liver tumor models. The experiments showed that PLA-5FU-SM5-1, PLA-5FU and 5-FU induced a 53.24%, 31.00%, and 18.11% tumor growth inhibition rate, respectively, on day 31 post-treatment determined by the bioluminescent intensity of the abdomen in tumor-bearing mice. Furthermore, we have calculated the three-dimensional location of the liver cancer in mice using a multilevel adaptive finite element algorithm based on bioluminescent intensity decay calibration. The reconstruction results demonstrated that PLA-5FU-SM5-1 inhibited the tumor rapid progression, which were consistent with the results of subcutaneous tumor mice experiments and in vitro cell experiment results.     

Noninvasive visualization of tumor growth in a human colorectal liver metastases xenograft model using bioluminescence in vivo imaging

Background

Bioluminescence imaging (BLI) is an ideal tool for noninvasive, quantitative monitoring of tumor progression/regression in animal models. The effectiveness of different treatment strategies is displayed by an altered intensity of bioluminescence, demonstrating a change of the tumor burden. The aim of this study was to establish a reliable, reproducible colorectal hepatic metastases cancer animal model.

Methods

Cells of the human colon carcinoma cell line HCT-116 Luc^pos expressing the firefly luciferase enzyme gene were used. HCT-116 Luc^pos cells (2.5 × 10⁶) were injected through the portal vein into the liver of immunoincompetent nude mice. BLI was used to analyze intrahepatic tumor burden and growth kinetic.

Results

HCT-116 Luc^pos cells demonstrated a progressive and reproducible growth in the liver after intraportal injection. Four days after injection, the animals were analyzed for tumor growth by BLI, and mice without or too low bioluminescence signals were excluded (between 10% and 20% animals). HCT-116 Luc^pos intrahepatic tumors responded successfully to different dosages (5 and 10 mg/kg) of 5-fluorouracil.

Conclusions

BLI is an important tool with many potential advantages for investigators. The measurement of intrahepatic tumor growth by imaging luciferase activity noninvasively provides valuable information on tumor burden and effectiveness of therapy. Thus, the presented intrahepatic metastases model based on the growth of HCT-116 Luc^pos cells is suitable for in vivo testing of different cancer therapy strategies.     

Bacterial adhesion to conventional hydrogel and new silicone-hydrogel contact lens materials

Background As bacterial adhesion to contact lenses may contribute to the pathogenesis of keratitis, the aim of our study was to investigate in vitro adhesion of clinically relevant bacteria to conventional hydrogel (standard HEMA) and silicone-hydrogel contact lenses using a bioluminescent ATP assay. Methods Four types of unworn contact lenses (Etafilcon A, Galyfilcon A, Balafilcon A, Lotrafilcon B) were incubated with Staphylococcus epidermidis (two different strains) and Pseudomonas aeruginosa suspended in phosphate buffered saline (PBS). Lenses were placed with the posterior surface facing up and were incubated in the bacterial suspension for 4 hours at 37°C. Bacterial binding was then measured and studied by bioluminescent ATP assay. Six replicate experiments were performed for each lens and strain. Results Adhesion of all species of bacteria to standard HEMA contact lenses (Etafilcon A) was found to be significantly lower than that of three types of silicone-hydrogel contact lenses, whereas Lotrafilcon B material showed the highest level of bacterial binding. Differences between species in the overall level of adhesion to the different types of contact lenses were observed. Adhesion of P. aeruginosa was typically at least 20 times greater than that observed with both S. epidermidis strains. Conclusions Conventional hydrogel contact lenses exhibit significantly lower bacterial adhesion in vitro than silicone-hydrogel ones. This could be due to the greater hydrophobicity but also to the higher oxygen transmissibility of silicone–hydrogel lenses. A minor part of this work was presented at the 2006 CLAO Congress.     

ATP生物荧光肿瘤体外药物敏感性检测在口腔癌化疗中的应用

目的探讨肿瘤体外药敏实验ATP生物荧光法(ATP-TCA)在口腔癌化疗中的应用。方法应用ATP生物荧光肿瘤体外药物敏感性检测法,检测48例口腔癌组织对6种常用化疗方案的敏感性。结果组织标本的可评估率为92.0%。口腔癌对TAT DDP最敏感,体外有效率为83.3%,其次为5-Fu DDP(75.0%),BLM(54.2%),DDP(60.4%),MTX(45.8%),5-Fu(62.5%)。化疗药物对口腔癌的杀伤作用具有较强的个体差异性。结论TP生物荧光肿瘤体外药物敏感性检测法敏感性高、稳定性好、简便、快速、检测结果可靠,可用于临床制定个体化的化疗方案。     

ATP生物发光技术快速检测水中细菌的研究

目的：探索利用ATP生物发光技术测定水中细菌量的可行性。方法：利用生物发光技术测定细菌中ATP量,根据细菌中ATP量与细菌数成正比这一原理,推算出水中菌落总数,与国标平皿计数法进行对比,分析两者之间的相关性。结果：检测结果与传统国标法有很好的相关性。结论：ATP生物发光技术具有准确、便利等优点,可望用于水中细菌的快速检测。     

Assessment of the environmental hazard from municipal and industrial wastewater treatment sludge by employing chemical and biological methods

Chemical analyses and toxicity testing were employed in conjunction to evaluate the environmental hazard from the wasted sludge generated during the biological treatment of urban and industrial wastewaters. Chemical analyses included determination of seven polychlorinated biphenyls (PCBs), 13 polycyclic aromatic hydrocarbons (PAHs), total organic carbon (TOC), and seven heavy metals (As, Cd, Cr, Cu, Pb, Mn, and Zn) in sludge and sludge leachates deriving from two standard leaching procedures: (a) the mild leaching test EN-12457-2 proposed by EC and (b) the relatively aggressive toxicity characteristic leaching procedure (TCLP) leaching test proposed by US EPA. Acute toxicity measurements were performed in aqueous sludge elutriates and leachates by using bioluminescence bacteria. The urban sludge was found to be more enriched with PAHs than the industrial sludge, however, at levels below the EU limits for sludge application. The total PCB content (Sigma7PCBs) in both sludges, particularly in the industrial sludge, exceeded the proposed European limit for sludge use as soil amendment. With regards to their heavy metal content, both sludges met the requirements for use in agriculture. The urban sludge exhibited high ecotoxicity, while the industrial sludge with almost two times lower toxicity was classified as not toxic to slightly toxic. The EN and the TCLP leaching procedures resulted in different sludge characterizations both from chemical and from ecotoxicological points of view. The EN procedure appeared to be more sensitive to the potential environmental risk from sludge disposal. The results of the study revealed the necessity for combining chemical with ecotoxicological criteria for integrated characterization of wasted sludge and the need for harmonization of the methods employed for waste classification.