小剂量X射线全身照射提高小鼠抗瘤能力的整体和离体研究

本文作者利用肿瘤血道转移和自发转移模型，从整体和离体两方面观察７５ｍＧｙＸ射线全身照射对小鼠抗瘤能力的影响。结果发现：①与假照组相比，照射组小鼠Ｂ１６黑色素瘤血道肺转移结节数明显降低（Ｐ＜０．０１）；过继转移至同系小鼠体内的脾细胞，可明显抑制Ｌｅｗｉｓ肺癌（ＬＬＣ）的局部生长和自发肺转移（Ｐ＜０．０５～０．０１）。②与假照组相比，体外检测照射组小鼠脾细胞对ＣｏｎＡＩＬ－２反应性增强（Ｐ＜０．０１），ＮＫ活性明显提高（Ｐ＜０．０１）；体内检测其巨噬细胞功能也显著增强（Ｐ＜０．０１），１２５ＩＵｄＲ标记黑色素瘤细胞在肺内的存留率显著降低（Ｐ＜０．０１）。上述结果提示，小剂量辐射可明显提高小鼠的抗瘤能力；机体免疫功能的增强可能是其抗肿瘤效应的主要机理之一。     

低剂量全身照射抑制小鼠癌细胞播散

小鼠受５０，７５，１００和１５０ｍＧｙＸ射线全身照射后２４小时经球后静脉注入Ｌｅｗｉｓ肺癌或Ｂ１６黑色素瘤细胞。注后１４天以计数肺肿瘤结节数为指标，发现受照小鼠癌细胞播散明显低于假照射对照小鼠。Ｌｅｗｉｓ肺癌细胞注入前２４小时接受７５ｍＧｙ全身照射小鼠与注入相同癌细胞数的假照射小鼠比较，发现照后２～６天脾脏ＮＫ细胞活性和ＩＬ－２分泌均增高。提示低剂量辐射可能通过增强免疫反应抑制癌细胞播散。     

低水平X射线照射对巨噬细胞吞噬消化功能的影响

本实验观察了不同剂量（０，５０，７５，１５０ｍＧｙ和２，４Ｇｙ）Ｘ射线单次全身照射昆明系小鼠后腹腔巨噬细胞（ＭΦ）对鸡红细胞（ＣＲＢＣ）吞噬消化功能的影响。其结果表明：５０和７５ｍＧｙ照射后３和５天巨噬细胞对ＣＲＢＣ的吞噬功能有非常明显增加（Ｐ＜０．０１），７５ｍＧｙ照射后巨噬细胞其消化功能也非常显著（Ｐ＜０．０１），而２和４Ｇｙ照射后其吞噬和消化功能降低。提示，低剂量辐射能够刺激机体非特异性免疫功能，大剂量照射使之受抑制。     

植瘤前后小剂量X线全身照射对小鼠肿瘤生长及转移的影响

目的：观察植瘤前后７．５ｃＧｙＸ线全身照射对肿瘤生长及转移的影响。材料与方法：将Ｃ５７ＢＬ／６小鼠随机分成三组：Ａ组（对照组）、Ｂ组（植瘤前２４小时照射组）和Ｃ组（植瘤后第７天照射组）。各组小鼠于足掌处注射Ｌｅｗｉｓ肺癌（ＬＬＣ）细胞悬液（每只鼠注射５×１０５个活细胞，０．１ｍｌ）。于植瘤后第２１天处死小鼠，检测各组小鼠ＬＬＣ原发瘤重量、自发肺转移结节数及转移率。结果：检测结果表明，Ｂ组ＬＬＣ原发瘤重量、自发肺转移结节数及转移率均明显低于Ａ组（Ｐ＜０．０５～０．００１）；Ｃ组原发瘤重量为Ａ组的５０％，肺转移结节数也显著降低（Ｐ＜０．００１）。Ｂ组的原发瘤重量为Ｃ组的２８％，肺转移结节数也较低（Ｐ＜０．０５）。于照射后２４小时检测小鼠免疫功能发现，照射后小鼠脾及胸腺有核细胞数增加，巨噬细胞功能增强，脾细胞的ＮＫ（自然杀伤细胞）活性及其对伴刀豆球蛋白（ＣｏｎＡ）和白细胞介素－２（ＩＬ－２）反应性均较未照射组明显提高（Ｐ＜０．０２～０．００１）。结论：上述结果提示，机体免疫功能的提高，可能是植瘤前后小剂量Ｘ线全身照射抗瘤效应的主要机制之一。     

小剂量X射线全身照射提高小鼠抗瘤能力的整体和离…

本文作者利用肿瘤血道转移和自发转移模型，从整体和离体两方面观察７５ｍＧｙＸ射线全身照射对小鼠抗瘤能力的影响。结果发现：①与假照组相比，照射组小鼠Ｂ１６黑色素瘤血道肺转移结节数明显降低（Ｐ〈０．０１）；过继转移至同系小鼠体内的脾细胞，可明显抑制Ｌｅｗｉｓ肺癌（ＬＬＣ）的局部生长和自发肺转移（Ｐ〈０．０５￣０．０１）。②与假照组相比，体外检测照射组小鼠脾细胞对ＣｏｎＡ ＩＬ－２反应性增强（Ｐ〈０．０１     

低剂量辐射适应性反应对辐射诱发小鼠胸腺淋巴瘤 …

目的 探讨低剂量对致癌剂量辐射诱发小鼠胸腺淋巴瘤的影响及其免疫学机理。方法 采用４次１．７５ＧｙＸ射线全身照射Ｃ５７ＢＬ／６Ｊ小鼠诱发胸腺淋巴瘤模型,观察不同剂量照后６个月小鼠胸腺淋巴瘤发生率,照后１个月脾脏ＮＫ细胞毒活性、ＩＬ－２和γ－ＩＦＮ分泌活性、腹腔巨噬细胞吞洚功能及其ＴＮＦ６分泌活性以及胸腺细胞分化的变化。结果 每次１．７５Ｇｙ照射前６ｈ、１２ｈ接受２５ｍＧｙ、７５ｍＧｙ全身照射均可降你     

低剂量X射线照射诱导G1期阻滞的适应性反应

目的观察低剂量Ｘ射线照射能否诱导Ｇ１期细胞阻滞。方法采用碘化丙啶（ＰＩ）荧光探针标记细胞，流式细胞术（ＦＣＭ）检测并分析细胞周期。结果证实１．５ＧｙＸ射线全身照射后１２小时，小鼠胸腺细胞发生明显Ｇ１期阻滞。而０．０７５Ｇｙ低剂量预照射可明显减轻其后１．５Ｇｙ攻击剂量所致Ｇ１期阻滞。离体照射ＥＬ－４淋巴瘤细胞也得到类似结果。结论０．０７５ＧｙＸ射线照射能诱导胸腺淋巴细胞及ＥＬ－４细胞Ｇ１期阻滞的适应性反应。     

OK-432对小鼠γ射线照射后T淋巴细胞功能的修复作用

对溶血性链球菌制剂ＯＫ－４３２促进照射小鼠Ｔ细胞功能的修复作用进行了探讨。方法实验用ＢＡＬｂ／ｃ小鼠，６０Ｃｏγ射线一次全身照射，吸收剂量５Ｇｙ。观察指标：小鼠脾淋巴细胞转化；ＩＬ－２产生及活性测定；迟发型超敏反应性测定。结果ＯＫ－４３２对几个Ｔ细胞亚群均具有作用，促进受照射小鼠对同种异型脾细胞的混合淋巴细胞反应及迟发型超敏反应的修复，促进脾脏中ＴＨ细胞产生ＩＬ－２，并能增强脾细胞对非特异性有丝分裂原的增殖反应。结论ＯＫ－４３２有促进多种Ｔ细胞亚群功能恢复的作用。     

Trolox对X射线照射后MOLT-4细胞凋亡蛋白酶-3依赖性凋亡的弱化作用

目的：用人白血病ＭＯＬＴ－４细胞系检测信号转导通路,研究照射后用Ｔｒｏｌｏｘ（一种抑制脂质过氧化作用的抗氧化剂）处理与Ｘ射线诱导的凋亡之间的关系。方法：用ＲＰＭＩ１６４０（含１０％胎牛血清）常规培养ＭＯＬＴ－４细胞,经７．５Ｇｙ（剂量率为１．８Ｇｙ／ｍｉｎ）Ｘ射线照射后用１０ｍｍｏｌ／ＬＴｒｏｌｏｘ处理５小时,通过台盼蓝排除实验、琼脂糖凝胶电泳及Ｗｅｓｔｅｒｎ印迹分别检测细胞存活率,ＤＮＡ片段化,ｐ５３、ＢＣＬ－２、ＢＡＸ的表达,激活的应激蛋白激酶／ｃ－ＪｕｎＮ－末端激酶（ＳＡＰＫ／ＪＮＫ）、凋…     

裂变中子照射小鼠脾脏NK活性和IL—2产生能力的变化

研究了裂变中子照射对小鼠脾脏ＮＫ活性和ＩＬ－２产生能力的剂量效应关系，并与γ线作了比较。裂变中子照后２４ｈＮＫ活性的量效曲线不典型：低剂量时明显下降，１．５－５．５Ｇｙ时高于正常，以后又下降。以Ｄ３７或Ｄ０为指标，中子相对生物学效应（ＲＢＥ）分别约为１．７和１．６。脾细胞ＩＬ－２产生能力对裂变中子和γ线的辐射反应相近，Ｄ３７均约为２．５Ｇｙ，裂变中子ＲＢＥ为１。     

Evaluation of chromosomal aberrations induced by 188Re-dendrimer nanosystem on B16f1 melanoma cells

Purpose: To study the rhenium-188 labeling of polyamidoamine (PAMAM) generation 4 (G4) dendrimer and its evaluation on biodistribution and chromosomal aberrations in melanoma cells induced by ionizing radiation as potential treatment agent.

Materials and methods: Dendrimers were first conjugated with Suc-HYNIC (succinimidyl 6-hydrazinopyridine-3-carboxylic acid hydrochloride). Dendrimer-HYNIC was then incubated with ¹⁸⁸ReO₄^?. Biodistribution was performed administrating ¹⁸⁸Re-dendrimer to normal (NM) or melanoma-bearing mice (MBM). Chromosome aberration test was conducted in order to measure treatment capacity of ¹⁸⁸Re-dendrimer in melanoma cells.

Results: Radiolabeling yield of dendrimer was approx. 70%. Biodistribution studies in NM showed blood clearance with hepatic and renal depuration. MBM showed a similar pattern of biodistribution with tumor uptake of 6% of injected dose. Aberrant metaphases quantified in control cells were 7%, increasing to 29.5% in cells treated with 15μCi (0.555 MBq) of ¹⁸⁸Re-dendrimer for 24?h.

Conclusions: ¹⁸⁸Re-dendrimer can produce double-stranded breaks in DNA induced by ionizing radiation in melanoma cells in vitro.     

Post-irradiation viability and cytotoxicity of natural killer cells isolated from human peripheral blood using different methods

Purpose We compared the pre- and post-irradiation viability and cytotoxicity of human peripheral natural killer cell (NK) populations obtained using different isolation methods.

Material and methods Three methods were used to enrich total NK cells from buffy coats: (I) a Ficoll-Paque gradient, plastic adherence and a nylon wool column; (II) a discontinuous Percoll gradient; or (III) the Dynal NK cell isolation kit. Subsequently, CD16⁺ and CD56⁺ NK cell subsets were collected using (IV) flow cytometry or (V) magnetic-activated cell sorting (MACS) NK cell isolation kits. The yield, viability, purity and cytotoxicity of the NK cell populations were measured using trypan blue exclusion, flow cytometry using propidium iodide and ⁵¹Cr release assays after enrichments as well as viability and cytotoxicity after a single radiation dose.

Results The purity of the preparations, as measured by the CD16⁺ and CD56⁺ cell content, was equally good between methods I–III (p?=?0.323), but the content of CD16⁺ and CD56⁺ cells using these methods was significantly lower than that using methods IV and V (p?=?0.005). The viability of the cell population enriched via flow cytometry (85.5%) was significantly lower than that enriched via other methods (99.4–98.0%, p?=?0.003). The cytotoxicity of NK cells enriched using methods I–III was significantly higher than that of NK cells enriched using methods IV and V (p?=?0.000). In vitro the NK cells did not recover cytotoxic activity following irradiation. In addition, we detected considerable inter-individual variation in yield, cytotoxicity and radiation sensitivity between the NK cells collected from different human donors.

Conclusions The selection of the appropriate NK cell enrichment method is very important for NK cell irradiation studies. According to our results, the Dynal and MACS NK isolation kits best retained the killing capacity and the viability of irradiated NK cells.     

急性放射病患者恢复期外周血淋巴细胞表型及功能分析

了解急性放射病患者恢复期间外周血淋巴细胞功能。方法用流式细胞仪分析淋巴细胞表型，３Ｈ－ＴｄＲ掺入法分别分析Ｔ细胞和ＮＫ细胞功能。结果照后４．５年，患者外周血ＣＤ＋４Ｔ细胞仍有不同程度低于正常对照；受照剂量大于２Ｇｙ者ＣＤ＋８Ｔ细胞数明显高于正常，而受照时年龄为５４岁者ＣＤ＋８Ｔ细胞明显低于对照；多数患者外周血ＮＫ细胞数和功能都高于正常对照。结论受照者外周血Ｔ细胞功能恢复与受照剂量大小及年龄因素有关，恢复期ＮＫ细胞数量和活性增高，对于弥补Ｔ细胞功能不足具有积极意义     

UVA radiation augments cytotoxic activity of psoralens in melanoma cells

Purpose: Melanoma is an aggressive form of skin cancer. The aim of the study was to evaluate the influence of UVA radiation and psoralens: 5-methoxypsoralen (5-MOP) or 8-methoxypsoralen (8-MOP) on melanoma cells viability.

Materials and methods: The amelanotic C32 and melanotic COLO829 human melanoma cell lines were exposed to increasing concentrations of psoralens (0.1–100 μM) in the presence or absence of UVA radiation. Cell viability was evaluated by the WST-1 assay.

Results: We demonstrated that 8-MOP, in contrast to 5-MOP, has no cytotoxic effect on both melanoma cell lines. Simultaneous exposure of cells to 8-MOP and UVA radiation caused significant cytotoxic response in C32 cells where the EC₅₀ value was estimated to be 131.0 μM (UVA dose: 1.3 J/cm²) and 105.3 μM (UVA dose: 2.6 J/cm²). The cytotoxicity of 5-MOP on both C32 and COLO829 cells was significantly augmented by UVA radiation – the EC₅₀ was estimated to be 22.7 or 7.9 μM (UVA dose: 1.3 J/cm²) and 24.2 or 7.0 μM (UVA dose: 2.6 J/cm²), respectively.

Conclusions: The demonstrated high cytotoxic response after simultaneous exposure of melanoma cells to psoralens and UVA radiation in vitro suggests the usefulness of PUVA therapy to treat melanoma in vivo.     

放射性药物99mTc-HL91在大鼠脑缺血模型的初步研究

目的 探讨国内首次合成的放射性药物＾９９ｍＴｃ－ＨＬ９１在脑血管疾病诊断中的应用的可能性。方法 对＾９９ｍＴｃ－ＨＬ９１进行一般性质、标记率、体外稳定性、异常毒性和正常小鼠体内生物分布等实验。建立１５只大鼠脑缺血模型,并在该模型上进行＾９９ｍＴｃ－ＨＬ９１体内分布和乏氧显像研究。结果（１）ＨＬ９１药盒标记简便,安全稳定,（２）正常小鼠静脉注射＾９９ｍＴｃ－ＨＬ－９１后血中放射性迅速下降,肝、肾和胃     

低剂量辐射对CIK细胞杀伤乳腺癌细胞作用的影响

目的 研究低剂量辐射对CIK细胞杀伤乳腺癌细胞MCF-7和SK-BR-3作用的影响,探讨其可能的作用机制。方法 取健康人外周血浓缩白细胞,常规分离出单个核细胞,加入不同的细胞因子培养7 d,诱导DC与CIK细胞,并将其按1 :5的比例共同培养7 d获得DC-CIK细胞。DC-CIK及CIK细胞分别给予40、80、120 mGy X射线照射,流式细胞仪检测细胞表型变化,运用CCK-8的方法检测其对乳腺癌细胞的杀伤作用。结果 DC-CIK组对乳腺癌细胞SK-BR-3较CIK组在不同效靶比均有更强的杀伤活性(t=3.63、5.62、8.21、5.49,P<0.05)。与CIK 0 mGy组相比,CIK细胞联合40、80、120 mGy低剂量辐射对乳腺癌细胞MCF-7和SK-BR-3的杀伤活性明显增高,而DC-CIK细胞联合低剂量辐射组与DC-CIK 0 mGy组相比,差异无统计学意义。结论 低剂量辐射对CIK细胞杀伤乳腺癌细胞有协同作用。     

153Sm-DTPA-c(CGRRAGGSC)对人MHCC97-H肝癌裸鼠模型的抑瘤作用

目的 探讨¹⁵³Sm-DTPA-c(CGRRAGGSC)对人MHCC97-H肝癌裸鼠模型移植瘤生长的抑制作用.方法 采用间接法合成¹⁵³Sm-DTPA-c(CGRRAGGSC)放射性药物.对8种不同细胞系IL-11受体表达,进行Western blot分析,选择肿瘤接种.20只MHCC97-H皮下荷瘤裸鼠按随机数字表法分为4组(对照组,低、中、高剂量组),每组5只.低、中、高剂量组尾静脉分别注入¹⁵³Sm-DTPA-c(CGRRAGGSC),剂量依序为5.5、11.0和22.0 MBq/0.2 ml,对照组尾静脉注入生理盐水0.2 ml.观察16 d后处死,比较肿瘤的体积,计算抑瘤率;观测标本病理改变;免疫组织化学检测5.5 MBq低剂量组瘤体组织治疗后Ki-67、Bcl-2和IL-11受体表达改变;Western blot检测不同剂量组IL-11受体的变化情况.结果 选择IL-11受体表达最高的MHCC97-H细胞接种.尾静脉内注射¹⁵³Sm-DTPA-c(CGRRAGGSC)后,早期瘤体中央可出现坏死、干瘪、结痂,后期又出现坏死周围瘤组织继续增长.低、中、高剂量组抑瘤率分别为(22.72±2.76)%、(34.65±2.36)%和(85.13±5.78)%(F＝89.32, P<0.05).免疫组织化学显示,对照组及低剂量组瘤内IL-11受体阳性率分别为(84.13±5.71)%和(61.57±5.98)%(t=13.62, P<0.05),低剂量组瘤细胞排列相对疏松,细胞内染色数量明显减少.低剂量组Ki-67、Bcl-2阳性率均较对照组下降(t=20.91、6.68, P<0.05).Western blot结果显示,随抑瘤作用增加,IL-11受体蛋白表达降低.结论 ¹⁵³Sm-DTPA-c(CGRRAGGSC)能够有效抑制人MHCC97-H肝癌裸鼠模型移植瘤的生长,促进IL-11受体表达下调及诱导凋亡作用.     

Testing ruthenium-based photoactivated chemotherapy on tumor-bearing mice models as a new treatment for uveal melanoma

INTRODUCTIONCurrently, there is no approved standard treatment for metastatic uveal melanoma and those approved for primary eye tumors are quite harmful to patients whose vision is often deteriorated or even lost^[1]. New clinical options are more than necessary. Because eyes are transparent to light and lasers are widely used in ophthalmology, photoactivated therapies represent an appealing modality for treating eye cancer. Light-activated technologies can be also used for liver metastasis through laparoscopy. Photoactivated chemotherapy (PACT) is a new light-activated technology combining the low systemic toxicity observed for photodynamic therapy (PDT), and the possibility of being activated in oxygen-poor tumors^[2,3].OBJECTIVESHere we investigated pre-clinically the relevance of ruthenium-based PACT compounds for anticancer therapies in the context of uveal melanoma.METHODSIn vitro experiments were conducted to determine the photocytotoxicity, the mode of cell death of these compounds and compare them to that of clinically used PDT compounds, both in normoxia and hypoxia environments. In vivo, biodistribution and anticancer efficacy studies were conducted using subcutaneous tumor models in mice, to investigate the best treatment scheme, e.g., drug-to-light interval, treatment dose, and light dose. Biosafety studies were also conducted through blood tests of drug-induced liver injury and acute kidney injury specific markers.RESULTSAltogether our results show promising apoptotic photocytotoxicity and a high antitumor efficacy, combined with rapid blood clearance and good biosafety in vivo.CONCLUSIONThese results in subcutaneous tumor models are promising and highlight the need for testing this new technology in clinically more relevant orthotopic tumor models in mice.     

Phase I/II 90Y-Zevalin (yttrium-90 ibritumomab tiuxetan, IDEC-Y2B8) radioimmunotherapy dosimetry results in relapsed or refractory non-Hodgkin’s lymphoma

Dosimetry studies in patients with non-Hodgkin’s lymphoma were performed to estimate the radiation absorbed dose to normal organs and bone marrow from ⁹⁰Y-Zevalin (yttrium-90 ibritumomab tiuxetan, IDEC-Y2B8) treatment in this phase I/II, multicenter trial. The trial was designed to determine the dose of Rituximab (chimeric anti-CD20, Rituxan, IDEC-C2B8, MabThera), the unlabeled antibody given prior to the radioconjugate to clear peripheral blood B cells and optimize distribution, and to determine the maximum tolerated dose of ⁹⁰Y-Zevalin [7.4, 11, or 15 MBq/kg (0.2, 0.3, or 0.4 mCi/kg)]. Patients received ¹¹¹In-Zevalin (indium-111 ibritumomab tiuxetan, IDEC-In2B8 ) on day 0 followed by a therapeutic dose of ⁹⁰Y-Zevalin on day 7. Both doses were preceded by an infusion of the chimeric, unlabeled antibody Rituximab. Following administration of ¹¹¹In-Zevalin, serial anterior/posterior whole-body scans were acquired. Major-organ radioactivity versus time estimates were calculated using regions of interest. Residence times were computed and entered into the MIRDOSE3 computer software program to calculate estimated radiation absorbed dose to each organ. Initial analyses of estimated radiation absorbed dose were completed at the clinical site. An additional, centralized dosimetry analysis was performed subsequently to provide a consistent analysis of data collected from the seven clinical sites. In all patients with dosimetry data (n =56), normal organ and red marrow radiation absorbed doses were estimated to be well under the protocol-defined upper limit of 20 Gy and 3 Gy, respectively. Median estimated radiation absorbed dose was 3.4 Gy to liver (range 1.2–7.8 Gy), 2.6 Gy to lungs (range 0.72–4.4 Gy), and 0.38 Gy to kidneys (range 0.07–0.61 Gy). Median estimated tumor radiation absorbed dose was 17 Gy (range 5.8–67 Gy). No correlation was noted between hematologic toxicity and the following variables: red marrow radiation absorbed dose, blood T _1/2, blood AUC, plasma T _1/2, and plasma AUC. It is concluded that ⁹⁰Y-Zevalin administered at nonmyeloablative maximum tolerated doses results in acceptable radiation absorbed doses to normal organs. The only toxicity of note is hematologic and is not correlated to red marrow radiation absorbed dose estimates or T _1/2, reflecting that hematologic toxicity is dependent on bone marrow reserve in this heavily pretreated population. Received 24 January and in revised form 20 March 2000     

榄香烯对小鼠脾淋巴细胞亚群及其调节的辐射保护效应

探讨榄香烯对受照小鼠脾淋巴细胞及亚群间调节作用的保护效应和剂量范围。方法应用单克隆抗体铺皿法分离受６０Ｃｏγ射线分次照射的小鼠脾脏Ｌ３，Ｔ４，Ｌｙｔ２和Ｂ细胞，分别测定榄香烯对以上三种亚群细胞和Ｌ３，Ｔ４，Ｌｙｔ２对Ｂ细胞３Ｈ－ＴｄＲ掺入的影响，同时测定ＳＯＤ活力和ＬＰＯ含量。结果榄香烯提高了受照２．５Ｇｙ（０．５Ｇｙ×５ｄ）小鼠脾淋巴细胞亚群的辐射抗性，增强了Ｌ３Ｔ４对Ｂ细胞的辅助调节作用，提高了脾脏ＳＯＤ活力和降低ＬＰＯ含量。而对受照４Ｇｙ（０．８Ｇｙ×５ｄ）和５Ｇｙ（０．５Ｇｙ×１０ｄ）组没有观察到这种效应。结论在一定剂量范围，榄香烯通过清除自由基减少靶分子的损伤。