锝-99m标记survivin反义寡核苷酸肝细胞癌显像的实验研究

目的研究放射性核素锝99m(99mTc)标记survivin反义寡核苷酸(ASON)显像诊断肝细胞癌的价值。方法用DNA合成仪合成18碱基单链survivinASON,在5′末端经氨基修饰后,以S乙酰基N羟基琥珀酰亚胺巯基乙酰基三甘氨酸(SacetylNHSMAG3)作为螯合剂对survivinASON进行99mTc标记,并对99mTcsurvivinASON在荷瘤(SMMC7721)裸鼠模型体内的生物学分布、肿瘤反义基因显像、肿瘤反义基因抑制显像进行了分析。结果肿瘤组织显像时间早,0.5h即已开始显影。99mTcsurvivinASON在肿瘤组织内的聚集程度随时间延长而逐渐增加,于4h时达到最大,肿瘤/对侧肢体肌肉比值分别为2.48±0.44(体外显像)和3.35±0.57(生物学分布)。正义寡核苷酸(SON)在肿瘤组织内的聚集各时间点均明显低于ASON,差异有统计学意义(P<0.01)。用未标记的survivinASON进行抑制后,99mTcsurvivinASON在肿瘤组织中的聚集明显减少,4h时的肿瘤/对侧肢体肌肉比值降为0.93±0.23,与抑制前的2.48±0.44相比有明显减低(P<0.01)。结论99mTcsurvivinASON可在荷瘤裸鼠模型的肿瘤组织中特异性聚集,为肝细胞癌的特异性诊断提供了一种可能的新方法。     

脂质体介导的^99mTc标记c—myc mRNA反义寡聚核苷酸链的生物学特性研究

目的：探索脂质体介导的^99mTc标记c-myc mRNA反义寡核苷酸链的生物学特性。方法：合成15bp的c-myc mRNA的反应，正义和无义寡核苷酸链，用三氯乙酸沉淀测定脂质体包裹的99mTc标的上述寡核苷酸链（简称为99mTc-DNA)和未用脂质体包裹的99mTc-DNA的血浆蛋白结合率。用BALB/c小鼠研究不同脂质体包裹的99Tc-DNA的生物学分布，用家兔研究脂质体介导的99mTc-DNA的药代动力学特性。结果：99mTc-DNA血浆蛋白结合率的范围为34．81％－70．53％，脂质体介导的99mTc-DNA的体内分布以网状内皮系统最高，胃，血和肠道其次，其余组织中放射性分 布较少，其药代动力学曲线符合开放二室模型，t1/2a约为2－5分钟，t1/2β约为100－150分钟，血浆清除率小于2ml/min，结论：99mTc-DNA的曲浆蛋白结合率高，脂质体介导的99mTc-DNA具有合适的生物半衰期，血浆清除块，是一种有开发潜力的放射性药物。     

99mTc-PDGFR-β AODN的制备及其生物分布

目的:寻求一种理想的99mTc标记大鼠血小板衍化生长因子受体-β(PDGFR-β)反义脱氧寡核苷酸(AODN)的方法,并观察标记产物的生物学分布以探讨其用于防治冠状动脉再狭窄的可能性. 方法: 先使长度为18个碱基的单链PDGFR-β AODN与双功能螯合剂NHS-MAG3耦联,然后进行99mTc标记,测定不同条件下标记率,分析确定最佳标记条件.常规测定放化纯度和比活度,并对标记物进行稳定性和正常小鼠体内分布实验. 结果: ①最佳标记条件下平均标记率达70%(最高为73.2%),经P4层析柱纯化后放化纯度>95%,比活度7.4～11.1MBq(200～300μCi)/ μg;② 99mTc-MAG3-AODN在室温下生理盐水中及37℃下新鲜人血清中稳定性良好,与血清蛋白结合率为6%～8%;③ 99mTc-MAG3-AODN在正常小鼠体内稳定性良好,在肾、肝中摄取较高. 结论: 以NHS-MAG3为鳌合剂标记得到的99mTc-MAG3-AODN具有良好的稳定性,为下一步进行细胞和动物实验提供了基础.     

脂质体介导的99mTc标记c-myc mRNA反义寡聚核苷酸链的生物学特性研究

目的探索脂质体介导的99m Tc标记c-m yc mRNA反义寡核苷酸链的生物学特性.方法合成15bp的c-myc mRNA的反义、正义和无义寡核苷酸链,用三氯乙酸沉淀测定脂质体包裹的99mTc标记的上述寡核苷酸链(简称为99mTc-DNA)和未用脂质体包裹的99mTc-DNA的血浆蛋白结合率.用BALB/c小鼠研究不同脂质体包裹的99mTc-DNA的生物学分布.用家兔研究脂质体介导的99mTc-DNA的药代动力学特性.结果 99mTc-DNA的血浆蛋白结合率的范围为34.81% ～7 0.53%.脂质体介导的99mTc-DNA的体内分布以网状内皮系统最高,胃、血和肠道其次,其余组织中放射性分布较少.其药代动力学曲线符合开放二室模型,t1/2α约为 2～5分钟,t1/2β约为100～150分钟,血浆清除率小于2ml/min.结论 99mTc-DNA的血浆蛋白结合率高,脂质体介导的99mTc-DNA具有合适的生物半衰期,血浆清除快,是一种有开发潜力的放射性药物.     

新型心肌灌注显像剂^99mTc—Q3的标记方法学研究

为获得高质量的心肌灌注显像剂,作者进行了９９ｍＴｃ－Ｎ,Ｎ'－亚乙基－二（乙酰丙 酮亚胺）二「三（３－甲氧基－１－丙基）膦」（＾９９ｍＴｃ－Ｑ３）的制备方法学研究。采用多元正交试验法进行了制备＾９９ｍＴｃ－Ｑ３最佳配方的筛选;用氯化亚锡化学还原法制备＾９９ｍＴｃ－Ｑ３;用柱沉析法进行＾９９ｍＴｃ－Ｑ３的分离和纯化、质量控制及体外稳定性试验;完成了无菌、热原、安全试验及家兔显像验证。制备９９ｍＴｃ－Ｑ     

脂质体介导的99mTc标记c-myc mRNA反义寡聚核苷酸链的生物学特性研究

目的　探索脂质体介导的 99m Tc标记 c- myc m RNA反义寡核苷酸链的生物学特性。方法　合成15 bp的 c- myc m RNA的反义、正义和无义寡核苷酸链 ,用三氯乙酸沉淀测定脂质体包裹的 99m Tc标记的上述寡核苷酸链 (简称为 99m Tc- DNA)和未用脂质体包裹的 99m Tc- DNA的血浆蛋白结合率。用 BAL B/ c小鼠研究不同脂质体包裹的 99m Tc- DNA的生物学分布。用家兔研究脂质体介导的 99m Tc- DNA的药代动力学特性。结果　 99m Tc- DNA的血浆蛋白结合率的范围为 34.81%～ 70 .5 3%。脂质体介导的 99m Tc- DNA的体内分布以网状内皮系统最高 ,胃、血和肠道其次 ,其余组织中放射性分布较少。其药代动力学曲线符合开放二室模型 ,t1 / 2α约为 2～ 5分钟 ,t1 / 2β约为 10 0～15 0分钟 ,血浆清除率小于 2 ml/ min。结论　 99m Tc- DNA的血浆蛋白结合率高 ,脂质体介导的 99m Tc- DNA具有合适的生物半衰期 ,血浆清除快 ,是一种有开发潜力的放射性药物     

99mTc标记葡萄糖99mTc-EC-DG在正常小鼠体内的分布研究

目的研究^99mTc标记葡萄糖99mTc-EC-DG在正常小鼠体内的生物学分布规律,用于临床肿瘤显像作基础研究.方法正常昆明小鼠48只,分8组,每组6只,实验前小鼠禁食8 h以上.尾静脉注射^99mTc-EC-DG 3.7 MBq (100 μCi) 后5 min、15 min、30 min、1 h、2 h、4 h、8 h和24 h放血处死,取心、肝、脾、肺、脑、肾、肌肉、骨、小肠、胃和血液等组织或器官,称重并测量其放射性,计算每克组织百分注射剂量率(%ID/g).结果^ 99mTc-EC-DG主要经肾脏代谢,脑不吸收^99mTc-EC-DG,肌肉组织摄取亦较少.各组织、器官的百分剂量率在1 h内除血液下降稍慢外,其余均有明显下降.结论^ 99mTc-EC-DG标记方便,体内外稳定性好,小鼠体内生物学分布表明其可能是一种较好的葡萄糖代谢显像剂.     

脂质体介导的99mTc标记c-myc mRNA反义寡核苷酸的反义作用效果的研究

目的探讨脂质体介导的99mTc标记癌基因c-myc mRNA的反义寡核苷酸能否抑制癌细胞的生长和癌蛋白的表达.方法合成15 bp的癌基因c-myc mRNA的反义、正义及无义寡核苷酸,用99mTc标记后(简称99mTc-DNA),一部分用脂质体包裹,以不同放射性强度的99mTc-DNA及脂质体包裹的99mTc-DNA转染细胞,于转染后不同时间测定细胞摄取率,在转染后18 h测定细胞返流比率,用四唑盐比色实验检测反义抑制细胞的生长状况,用流式细胞术测定癌蛋白的表达.结果在1～5 h内,细胞的摄取率随时间的延长而增加,脂质体包裹的99mTc-DNA的细胞摄取率明显高于未用脂质体包裹的99mTc-DNA.四唑盐比色实验,随着脂质体介导的99mTc-反义DNA剂量的增加,细胞数量逐渐减少,而正义、无义寡聚核苷酸组,细胞数量则没有减少的趋势.细胞的返流比率,反义、正义、无义寡核苷酸分别为39.51%、44.12%、63.92%.测定癌蛋白的荧光强度,反义寡核苷酸组2.9860±0.3733、正义寡核苷酸组4.2600±0.2218、无义寡核苷酸组5.2620±0.8562,反义寡聚核苷酸组的荧光强度明显低于正义和无义寡聚核苷酸组.结论脂质体介导的99mTc标记的反义寡核苷酸能抑制癌细胞的生长和癌蛋白的表达.     

99mTc标记c-erbB2反义寡脱氧核苷酸对乳腺癌显像的实验研究

用放射性核素标记癌基因反义寡脱氧核苷酸（antisense oligodeoxynucleotides，ASODN）对肿瘤进行显像，可以探测只有癌基因激活、扩增和过度表达还没有形成实体的肿瘤组织，以实现对肿瘤的早期诊断。乳腺癌是妇女发病率和死亡率最高的主要恶性肿瘤，如果能在肿瘤早期鉴别其癌基因的表达类型及临床分期，施行“量体裁衣”式的治疗，可提高乳腺癌治疗的效果。c-erbB2是乳腺癌原癌基因，与乳腺癌的恶性转化和不良预后密切相关。     

反义c-myc寡核苷酸对半乳糖诱导的晶体上皮细胞生长的抑制作用

目的探讨反义c-myc寡核苷酸(ASODN)对半乳糖诱导的兔晶体上皮细胞(LEC)生长的影响.方法人工合成与c-myc基因第二外显子翻译起始区序列互补的寡核苷酸,用半乳糖和反义寡核苷酸处理培养的晶体上皮细胞,应用细胞计数法和MTT法观察反义c-myc寡核苷酸对半乳糖诱导的晶体上皮细胞增殖的影响.结果 2.5～10.0 μmol/L反义c-myc寡核苷酸能抑制半乳糖诱导的晶体上皮细胞的增殖,10.0 μmol/L时作用较显著.结论反义c-myc寡核苷酸对半乳糖诱导的白内障晶体上皮细胞的生长增殖有抑制作用.     

99mTc螯合两种抗肝癌单链抗体的标记条件

目的:研究两种全新的抗肝癌基因工程单链抗体hdsFv和scFv-PE38的99mTc标记方法及适宜条件以应用于放免显像. 方法:选用二乙烯三胺五乙酸环酐(cDTPAa)作为螯合剂,沿用比较成熟的标记方法,以标记率和标记物免疫活性为指标分别测定两种单链抗体与99mTc偶联的适宜条件,并测定标记物的比活度和体外稳定性. 结果:其他理化条件不变,cDTPA与hdsFv的浓度比为20: 1～40: 1,99mTc活度37～148 MBq时,hdsFv标记率为62%～84%;cDTPA对scFv-PE38的浓度比为20: 1～100: 1,99mTc活度为37～148 MBq时,scFv-PE38标记率为68%～89%;且两种标记单链抗体的免疫活性均在60%以上. 结论:通过对这两种全新的单链抗体的标记和检测,得到了尽可能保持其原有免疫活性的标记条件,为通过放免显像在活体内检验单链抗体亲瘤性奠定了基础.     

99mTc甲氧基异丁基异腈定位显像诊断甲状旁腺功能亢进症

目的：评价^99mTc甲氧基异丁基异腈（MIBI）双时相显像法对甲状旁腺功能亢进（甲旁亢）的诊断价值。方法：21例临床诊断甲状旁腺功能亢进患者，静注注入T99mTc MIBI 740MBq（20mCi），分别于10～20min（早期显像）、2h（延迟显像）采集颈部至上胸部的前位平面像。结果：5例原发性甲旁亢患者均表现为孤立的放射性浓聚灶；16例继发性甲旁亢中，2例显示为单个放射性浓聚灶，10例显示为2个放射性浓聚灶，2例显示为3个放射性浓聚灶；另有2例显示有异位甲状旁腺病灶。10例经手术证实甲旁亢患者中，术前^99mTc MIBI甲状旁腺显像和超声诊断检查，对原发性甲旁亢患者定位诊断阳性率均为100％，对继发性甲旁亢患者定位诊断阳性率分别为55％和70％。结论：^99mTc MIBI双时相法显像在甲旁亢患者中有较高的定位诊断价值。     

新型心肌灌注显像剂99mTc-Q3的临床前药理研究

为了给临床应用研究提供依据 ,作者对新型心肌灌注显像剂 99m Tc- N,N′-亚乙基 -二 (乙酰丙酮亚胺 )二〔三 (3-甲氧基 - 1 -丙基 )膦〕(99m Tc- Q3)进行了临床前药理研究。其内容包括 :99m Tc- Q3在兔体内的血药清除动力学、血浆蛋白结合率、小鼠体内分布、家犬显像及异常毒性试验等。结果显示 :1 9m Tc- Q3的放化纯度 >99% ,室温下放置 6小时稳定 ;2兔血药清除动力学符合一次静脉给药的药代动力学二室模型 ;3小鼠体内分布显示 99m Tc- Q3在小鼠心肌摄取早 ,摄取值高 ,且放射性滞留时间长 ,血、肺及肝药物清除均快 ;4从家犬显像可见 ,99m Tc- Q3在肺中清除迅速 ,1小时后肺部放射性水平接近本底 ,1 5分钟时心肌影像已清晰可见 ,3小时后才逐渐消退 ;5 99m Tc- Q3的兔血浆蛋白结合率低 (7.1 3± 0 .42 % ) ;6小鼠对药物的耐受剂量为人的 5 0 0倍。上述结果表明 ,99m Tc- Q3的体外性质稳定 ,心肌摄取早 ,摄取率高 ,且放射性滞留时间长 ,血、肺及肝清除快 ,安全 ,具有临床研究价值     

The value of 99mTc methylene diphosphonate bone scintigraphy in diagnosing relapsing polychondritis

Relapsing polychondritis （RP） is a recurrent disease involving cartilage mainly of the ear,nose, larynx, trachea, and bronchus. The typical manifestations of the disease in the ear and nose can be easily recognized, but the symptoms could be ignored or easily confused with those of other diseases when the cartilage of other sites is involved. Thus, it is necessary to develop a new technique for the diagnosis of this disease. Few cases of abnormal accumulation of radioactivity at cartilage shown by ^99mTc methylene diphosphonate （MDP） bone scintigraphy are described in the literature. In this report, we present 4 patients of whom 3 had positive findings on ^99mTc MDP bone scintigraphy with an assessment of ^99mTc MDP bone scintigraphy in the diagnosis of RP.     

摄~(99m)锝率测定快速诊断甲状腺功能的临床应用研究

目的探讨摄99m锝率快速诊断甲状腺功能的临床应用价值。方法用核多功能仪分别对49例甲状腺功能亢进症未服药者(甲亢组)、13例甲亢服药者(甲亢服药组)、9例亚急性甲状腺炎者(亚甲炎组)以及18例正常健康体检者(对照组)进行甲状腺摄131碘率和摄99m锝率检测。结果甲亢组、甲亢服药组其甲状腺摄99m锝率在第30、60min时高于正常对照组,而亚甲炎组低于正常对照组。甲亢组摄131碘率在第2h、6h和24h高于正常对照组,甲亢服药组与对照组差别不大,而亚甲炎组低于正常对照组。结论摄99m锝率与摄131碘率均可反映甲状腺的功能状态,摄99m锝率检查可快速诊断,方便患者,是测定甲状腺功能的一种实用、简便又敏感的方法。     

益气养阴祛瘀法对干燥综合征患者^99mTc04^-腮腺显像及免疫功能的影响

目的：探究益气养阴祛瘀法对原发性干燥综合征(PSS)患者^99mTc04^-腮腺显像及免疫功能的影响。方法：选择2012年6月-2015年6月在我院接受治疗的PSS患者120例,采用随机对照观察研究方法,将入选的患者随机分为对照组与治疗组,每组各60例,对照组行中药白芍总苷胶囊治疗,治疗组在对照组基础上加服益气养阴祛瘀汤治疗,并选择同期就诊的健康体检者60例作为正常组,对比不同组别之间的临床症状、腮腺功能、免疫功能及不良反应的差异。结果：治疗后6个月,两组的中医证候积分,腮腺摄取、排泌功能,IgA、IgG、IgM含量,抗SSA、抗SSB阳性率,CD4、CD4/CD8比值均有显著改善,其中治疗组改善幅度优于对照组(P＜0.05),有统计学意义。结论：益气养阴祛瘀法能改善PSS患者临床症状,提高腮腺摄取、排泌功能,并能调节机体免疫功能。     

99m Tc-MAG3-ASON的制备及其在荷乳腺癌裸鼠中的分布

目的:探讨99mTc标记的反义寡核苷酸(ASON)的制备及其在荷乳腺癌裸鼠中的分布.方法:一步法合成巯基乙酰三甘氨酰-N-羟基琥珀酰亚胺酯(NHS-MAG3),并与c-erbB2 mRNA互补的5'末端氨基修饰的15个碱基的ASON偶联;随后进行99mTc的标记,并用SephadexG25分离纯化99mTc-MAG3-ASON,并评价其稳定性;最后检测其在荷乳腺癌BALB/c裸鼠体内的分布.结果:99mTc-MAC3-ASON平均标记率为70.6%;纯化后,在室温下放置4h,其放化纯度为94.3%;与人血清孵育后,其放化纯度为93.8%;与血浆蛋白结合率为11.8%.乳腺癌组织的摄取率2h达峰值(6.09% ID/g).结论:以NHS-MAG3为螯合剂制备的99mTc-MAG3-ASON具有良好的稳定性,在乳腺癌部位高浓聚,可望用于肿瘤的显像诊断和治疗.     

Pre-clinical evaluation of a new indirectly labeled 99mTc-6-hydrazinopyridine-3-carboxylic acid (HYNIC)-depreotide with HYNIC as bifunctional chelator

Background  Technetium-99m or ^99mTc is widely used for labeling peptide in nuclear medicine. Somatostatin and its analog can inhibit tumor cell growth after binding with its receptor. This research was to study the preclinical effect of a new ^99mTc-6-hydrazinopyridine-3-carboxylic acid (HYNIC)-depreotide, indirect ^99mTc labeling of depreotide using HYNIC as a bifunctional chelator.

Methods  The cyclopeptide, cyclo-[(N-Me) Phe-Tyr-D-Trp-Lys-Val-Hcy], the linear peptide, and [ClCH₂-CO×b-Dap-Lys- Cys-Lys×amide] were synthesized by Fmoc solid-phase synthesis. The cyclopeptide and the linear peptide were linked by liquid-phase synthesis. The product depreotide was isolated and purified by high performance liquid chromatography and was confirmed by mass spectrography. Depreotide was labeled with ^99mTc through a direct labeling method, using HYNIC as a bifunctional chelator. Paper chromatography method was used to calculate the labeling rate, and through the comparative analysis selected the best mark conditions. The new ^99mTc-HYNIC-depreotide was tested by high-performance liquid chromatography (HPLC). The internalization and externalization rates of the new ^99mTc-HYNIC-depreotide were studied in A549 cells. Furthermore, biodistribution of the radiopeptide was studied in nude mice, bearing tumors from human lung carcinoma cells SPC-A1.

Results  The molecular of synthesize depreotide was 1358, and the purity of it was 95.29%. The labeling efficiency of ^99mTc-HYNIC-depreotide was highest at pH 6.0 and 15°C, about (70.95±0.84)%. The labeling rate of the new ^99mTc-HYNIC-depreotide rose to a peak of (20.75±0.48)% at 60 minutes in A549 cells at 37°C and decreased slightly later, while it elevated gradually during the time course at 4°C and 25°C. The internalization rate of the new ^99mTc-HYNIC-depreotide at 37°C increased gradually and reached the peak of 84.4% in 120 minutes, while the externalization rate of the new ^99mTc-HYNIC-depreotide was always less than 20%. In mice bearing the experimental SPC-A1 tumor, the new ^99mTc-HYNIC-depreotide demonstrated a high tumor uptake of (4.05±0.04)% ID/g at 1.5 hpi and remained high ((2.51±0.06)% ID/g) at 4 hpi. The tumor-to-lung activity concentration ratio (T/Lu) was very high for the new ^99mTc-HYNIC-depreotide at all time points. So did the tumor-to-muscle activity (T/Mu) and tumor-to-blood activity concentration ratios (T/Bl).

Conclusion  The findings suggested that the new ^99mTc-HYNIC-depreotide might be a promising candidate radiopharmaceutical for imaging somatostatin receptor positive lung cancer.     

^99mTc-YIGSR as a Receptor Tracer in Imaging the Ehrlich Ascites Tumor-bearing Mice as Compared with ^99mTc-MIBI

The validity of ^99mTc-YIGSR, a novel receptor radio-tracer, in imaging the Ehrlich ascites tumor was evaluated. YIGSR, a pentapeptide of laminin, was labeled with ^99mTc by using a bifunctional chelator S-Acetly-NH3-MAG3. The MIBI was labeled with ^99mTc by following the kit instruction. The mice of tumor group were intravenously injected 1-2 mCi of ^99mTc-YIGSR or ^99mTc-MIBI via caudal vein, immobilized and imaged under a Gamma camera. The same procedure was performed in mice of blockade group, in which the unlabeled YIGSR was previously injected to block the receptor-recognition sites, and inflammation group serving as control. The reverse-phase Sep-Pak C18 chromatogram was found to have an essentially complete conjugation between YIGSR and S-Acetly-NH3-MAG3. The conjugated YIGSR could be radio-labeled successfully with ^99mTC at room temperature and neutral pH, with a radio-labeling yield of 62%. Without the chelator S-Acetly-NH3-MAG3, the YIGSR was labeled with ^99mTc at an efficiency of 4%. The imagological study revealed obvious tumor accumulation of ^99mTc-YIGSR 15 min after the injection, and the uptake peaked after 3 h with a tumor-to-muscle ratio （T/M） of 11.36. The radio-tracer was slowly cleared up and resulted in a T/M of 3.01 at the 8th h after the injection. As for blocked group, the tumor uptake of radiotracer was significantly lower, with the highest T/M being 4.61 after 3 h and 0.89 after 8 h. The T/M was 3.72 at the 3rd h and 1.29 at the 8th h after the ^99mTc-YIGSR injection in the inflammatory group. The T/M was significantly higher in tumor group than in inflammatory group or control group （P〈0.001）. In the ^99mTc-MIBI group, the T/M was 1.40 at the 3rd h and 0.55 at the 8th h after the injection, which showed a significant difference as compared with ^99mTc-YIGSR （P〈0.001）. It is concluded that YIGSR can be successfully radiolabelled by using S-Acetly-NH3-MAG3. ^99mTc-YIGSR has many advantages in tumor imaging, such as quick and clear visualization, high sensitivity and sp