99mTc通过双功能螯合剂HYNIC间接标记depreotide的临床评价研究

Objectives: Technetium-99m or 99mTc is widely used for labeling peptide in nuclear medicine. Somatostatin and its analogu can inhibit tumor cell growth after biding with its receptor. This research was to study the preclinical effect of a new 99mTc-HYNIC-depreotide, indirect 99mTc labeling of depreotide using HYNIC as bifunctional chelator. Methods: The cyclopeptide , cyclo-[(N-Me) Phe-Tyr-D-Trp-Lys-Val-Hcy ] , the linear peptide , and [ ClCH2-CO.β-Dap-Lys-Cys-Lys. amide ] were synthesised 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 bifunctional chelator. Used the paper chromatography method to calculate 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 efficient of 99mTc-HYNIC-depreotide was highest in 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 min in A549 cells at 37 °C and just slightly decreased later, while it elevated gradually during 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 min, 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 a remained high one of 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 ratio (T/Bl). Conclusion: The findings suggested that the new 99mTc-HYNIC-depreotide might be a promising candidate radiopharmaceutical for imaging SSTR positive lung cancer. Keywords: Technetium-99m; HYNIC; depreotide; lung carcinoma;     

基于VEGFR靶点的小分子肿瘤靶向抑制肽的99Tcm标记及鉴定

目的 探讨基于血管内皮生长因子受体( vascular endothelial growth factor receptor,VEGFR)靶点的小分子肿瘤靶向抑制肽理想的放射性核素99 Tcm标记方法.方法 多肽QKRKRKKSRKKH、RKRKRKKSRYIVLS分别经双功能螯合剂S-acetyl-MAG3、GA(D)GG、HYNIC修饰后进行放射性核素99 Tcm标记,HPLC及纸层析法鉴定其标记率及放化纯度,3 H-TdR参入实验鉴定其抑制细胞增殖的能力,体外受体竞争结合实验鉴定其受体结合活性.结果 以HYNIC为双功能螯合剂,这2条多肽的99 Tcm标记率分别为(94.13±1.77)％、(93.79±3.53)％,明显高于S-acetyl-MAG3、GA(D) GG修饰多肽的标记率.经HYNIC修饰后的多肽对肿瘤细胞A549增殖的抑制率约为(32.86±3.36)％、(61.50±4.50)％,与修饰前无明显变化(P＞0.05);多肽标记后仍保持了良好的VEGFR结合活性,室温下放置24 h后,放化纯度仍高达90％左右.结论 以HYNIC为双功能螯合剂,以TPPTS为协同配体及还原剂,多肽QKRKRKKSRKKH、RKRKRKKSRYIVLS的99Tcm标记率高,且仍具有良好的抑制肿瘤细胞增殖的能力和VEGFR结合活性.     

CEA单克隆抗体C50的99mTc标记试剂盒的初步应用

目的进行癌胚抗原CEA单克隆抗体C50的99mTc标记试剂盒的初步应用。方法进行99mTc-C50试剂盒的安全限度检测,进行99mTc-C50试剂盒的动物实验应用研究,进行3例结直肠癌患者的RII。结果 (1)注射试剂后3只家兔的体温升高值均在规定范围内,5只受试小鼠注射试剂后48 h内无一死亡;(2)荷瘤裸鼠RII,药物注入后6 h肿瘤部位有放射性聚集,24 hT/NT比值为3.46～4.68,24 h后,经活体组织测量发现瘤体与血液的T/NT比值为1.84,与肌肉的比值高达16.1;(3)3例大肠癌患者中复发、转移灶均显示出阳性结果,T/NT比值在4.68～12.36之间。结论本标记化合物无热源性,无毒性,符合国家药典的规定,为99mTc-C50能较好地应用于临床研究提供依据。     

肾性继发性甲状旁腺功能亢进99mTc⁃MIBI显像的影响因素分析

目的：探究肾性继发性甲状旁腺功能亢进(secondary hyperparathyroidism,SHPT)^99m 锝-甲氧基异丁基异腈(^99m Tc-MIBI) 显像的相关影响因素。方法：回顾性分析在南京医科大学第一附属医院接受手术治疗的482例肾性SHPT患者的临床资料,以术后病理为对照,比较不同显像方案(A组：仅行^99m Tc-MIBI双时相法平面显像;B组：在A组显像基础上加做早期相SPECT/CT) 对SHPT诊断效能的差异。根据影像与病理结果是否一致,再将B组患者分为完全诊断组(MIBI诊断结果与病理结果完全一致)与部分诊断组(MIBI诊断的腺体数目少于病理结果),进一步分析临床特征和生化指标对MIBI显像的影响。结果：在SHPT 诊断效能分析中,B组的灵敏度、特异度、准确度和阳性预测值均高于A组(81.9% vs. 70.3%;97.1% vs. 60.0%;82.2% vs. 69.9%; 99.9% vs. 97.9%,P ＜ 0.01),但两组阴性预测值差异无统计学意义(6.8% vs. 10.2%,P ＞ 0.05)。多因素Logistic回归分析示,血清磷离子水平是预测MIBI显像假阴性的独立危险因素(OR=0.57,95%CI：0.372~0.866,P = 0.007)。ROC曲线显示,当血清磷离子水平 ＜ 1.97 mmol/L时,^99m Tc-MIBI 双时相法平面显像加早期相SPECT/CT诊断SHPT发生漏诊的风险增加。结论：^99m Tc-MIBI 双时相法平面显像加做早期相SPECT/CT可提高SHPT患者功能亢进甲状旁腺腺体的检出灵敏度及准确度。影像结果的判断应当结合血清磷离子水平,当血清磷离子水平较低时,MIBI显像存在漏诊的可能,影像医生和临床医生都须予以关注。     

99mTc一MDP骨显像中双肺弥漫显影2例

99mTc-MDP骨显像中双肺弥漫显像者较为少见,笔者遇到2例,报道如下.     

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.     

99mTc标记的新型肺癌多肽分子探针及其生物学分布

目的探讨99mTc标记特异性靶向肺癌小分子多肽的标记方法,观察99mTc-多肽在正常动物体内的动力学特性及体内生物学分布特点。方法采用NHS-MAG3双功能螯合法进行99mTc标记小分子多肽(cNGQGEQc),纸层析法测定标记率,通过体外稳定性实验、半胱氨酸置换实验及血清蛋白结合实验评价99mTc-多肽的放化特性,测定标记物静脉注射后在新西兰大白兔不同时相血液放射性清除曲线;分别测定经尾静脉注射7.4 MBq标记物后的小鼠体内各脏器质量数和放射性计数,计算各脏器每克组织百分注射剂量率(%ID/g);经耳缘静脉注射37 MBq标记物,应用SPECT显像法观察标记物在大白兔体内的放射性动态分布变化。结果99mTc标记多肽的标记率>90%,室温下和血清中37℃下放置12 h放化纯度分别为91%和85%;半胱氨酸置换率<7%;血清蛋白结合率<5%。99mTc标记多肽在兔体内血放射性清除迅速;在小鼠体内清除较快,主要通过肾脏排泄,其余脏器内放射性均较低(P<0.05)。结论99mTc标记基于肺癌特异性靶向多肽的标记方法简单,标记率高,具有良好的体内外稳定性,具有比较理想的体内分布动力学特性。     

99mTc-CL3-Bt的制备、质控及生物分布研究

目的制备     

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.     

双功能多普勒对高血压病患者心内结构及心功能变化的研究

研究目的揭示高血压病患者心内结构及功能变化特点。研究背景作者在工作中发现高血压病患者心内结构及功能较正常人有许多不同之处。研究方法应用双功能多普勒仪,测量高血压病患者心内结构（左房、左室大小,左室后壁、室间隔厚度,主动脉辩损害,主动脉辩返流）、左室功能（左室村血分数EF、左室舒张早期血流峰值流速PFVE、房缩期最大血流速度PFVA及PFVA／PFVE）,并设立正常对照组。经统计学处理判断组间各项指标的差异。结果高血压病患者左房改正常人增大,左室后壁及室间隔被正常人增厚（P＜0．01）,主动脉辩损害及返流较正常人增多,PFVE降低,PWA增高,PWA／PFVE增大（P＜0.01）。结论高血压病患者外周阻力的增高导致心内结构及功能的一系列改变。     

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。     

不同病程老年2型糖尿病降糖药物应用及分析

目的2型糖尿病不同病程阶段使用的降糖药物不同,探讨随着胰岛p细胞功能下降,降糖药物用量是否应持续增加。方法收集126例老年2型糖尿病患者,病程每5年为一个阶段分成四组,统计每纽降糖药物用量,分析病程对降糖药物用量的影响。结果病程10年以下者口服降糖药物用量最大,病程5年以下者二甲双胍用量最大。病程10年以上者胰岛素用量明显增多,但病程继续延长,降糖药物用量无需增加。结论对病程10年以上的老年糖尿病患者需要的降糖药物不会继续增长,应警惕低血糖发生。     

(99m) ~ Tc-ccM4嵌合抗体肺癌放射免疫显像初步临床应用

目的：研究９９ｍ Ｔｃ ｃｃ Ｍ ４嵌合抗体用于肺癌放射免疫诊断。方法：用直接法９９ｍ Ｔｃ 标记嵌合抗体 ｃｃ Ｍ ４对１７例疑肺癌患者行放射免疫显像（ Ｒ Ｉ Ｉ）,同时用１２５Ⅰ标记 ｃｃ Ｍ ４检测患者血清中的 Ｔ Ａ Ｇ ７２抗原水平。显像后２周检测病人血清中的人抗鼠抗体（ Ｈ Ａ Ｍ Ａ）水平。结果：肺癌患者中 Ｔ Ａ Ｇ ７２抗原水平明显高于正常对照者（ Ｐ＜ ００５）,阳性率为４６４％ 。注射９９ｍ Ｔｃ ｃｃ Ｍ ４后１２ ｈ 癌灶对心肝的靶／非靶比值分别为２６６和２０８,选取此时的显像结果与手术及病理对比,灵敏度为７２２％ （１３／１８）,对于１４个原发灶 Ｒ Ｉ Ｉ示１１个有放射性浓集,灵敏度为７８６％ ,纵隔转移灶３个中２个 Ｒ Ｉ Ｉ示放射性浓集,１个肝转移灶阴性。３例非癌性病变未见放射性浓集。注射抗体２周后病人血清中 Ｈ Ａ Ｍ Ａ 均未见阳性反应。结论：９９ｍ Ｔｃ ｃｃ Ｍ ４肺癌 Ｒ Ｉ Ｉ诊断原发性肺癌是一种可行的方法。     

心力衰竭患者阵发性心房颤动时对四种不同抗心律失常药治疗的反应

目的 :对充血性心力衰竭患者合并阵发性心房颤动 (简称房颤 )时观察四种不同的抗心律失常药静脉注射后的疗效。　方法 :选择心力衰竭合并阵发性房颤患者 5 5例 ,心功能NYHA分级达到Ⅱ、Ⅲ级 ,阵发性房颤发作时心室率在 12 0次 /min以上。将患者随机分为四组 ,分别给予毛花苷丙、胺碘酮、地尔硫 艹卓 、艾司洛尔静脉注射 ,观察用药前后心室率、心律、血压等指标的变化。　结果 :地尔硫 艹卓 和艾司洛尔组用药后 1h内心室率均有明显下降 ,而毛花苷丙和胺碘酮组心室率无变化 ,四组 2 4h内房颤转复成功率无差异 ,用药前后血压亦无明显变化。　结论 :充血性心力衰竭患者发作阵发性房颤时 ,地尔硫 艹卓 和艾司洛尔减慢心室率的疗效优于毛花苷丙和胺碘酮     

L-LWiDose4技术在新生儿先天性心脏病CT血管成像中的临床应用

目的：探讨低X线辐射剂量及低碘浓度等渗对比剂联合iDose4迭代重建技术（L-LWiDose4）在新生儿先天性心脏病CT血管成像（CTA）中的可行性。方法：前瞻性收集2015年1月至2018年3月温州医科大学附属第二医院育英儿童医院新生儿科需行心脏CTA检查的连续102例患者,随机分为2组,采用不同的扫描方案：常规组（A组）53例,采用100 kVp、100 mAs、碘海醇（350 mg I/mL）,重建方式为滤波反投影（FBP）算法;实验组49例,采用80 kVp、80 mAs、碘克沙醇（270 mg I/mL）,根据iDose4迭代算法可以得到iDose4-1～iDose4-7 7组图像,选取iDose4-2～iDose4-6 5组图像,分别记为B组、C组、D组、E组、F组。依据新生儿体质量采取不同的对比剂注射剂量并计算碘摄入量。测量主动脉弓、气管分叉水平降主动脉、肺动脉干及主动脉弓层面两侧胸大肌和竖脊肌CT值,测量气管分叉层面周围空气CT值的标准差（SD）,记为背景噪声,计算信噪比（SNR）和对比噪声比（CNR）,并对图像质量进行主观评价。记录容积CT剂量指数（CTDIvol）和剂量长度乘积（DLP）,并计算有效剂量（ED）和体型特异性剂量评估（SSDE）。以手术或心导管造影为“金标准”,统计并比较各组CTA检查的诊断符合率。结果：实验组CTDIvol、SSDE、DLP、ED较常规组分别降低了57.08%、56.62%、58.31%、56.97%（P＜0.05）。常规组、实验组对比剂碘摄入量分别为（1.83±1.27）g和（1.61± 1.36）g,差异有统计学意义（P＜0.05）。B组、F组图像客观评价不及A组、C组、D组、E组（P＜0.05）。A组、D组、E组图像主观评价差异无统计学意义。常规组、实验各组诊断符合率分别为98.6%、81.6%、87.7%、97.9%、97.9%、96.9%,差异无统计学意义（P＞0.05）。结论：80 kVp、80 mAs、碘克沙醇（270 mg I/mL）联合iDose4迭代重建技术新生儿先天性心脏病CTA检查,在降低患者的电离辐射剂量和对比剂碘剂量的同时,其图像质量能够满足临床诊断需要。     

99mTc-MIBI心肌灌注断层显像评价心肌梗死后左室重构

目的：压用^99mTc-甲氯异丁基异腈（MIBI）心肌灌注显像对左室重构进行半定量评估。方法：随机从^99mTc-MIBI心肌灌注显像库中抽取心肌梗死患者55例．其中前壁组25例，下后壁组30例；正常对照组30例。采用自编软件测定前壁或下后壁梗死部位局部膨出指标EG／EF或FG／EF值、左室心肌梗死后球形化指标EF／AB值．结果：前壁组EG/EF值明显高于正常组．下后壁组FG／EF值与正常组无明显差别．前壁组、下后壁组EF/AB值均明显高于正常组．前壁组EF／AB值明显高于下后壁组．结论：心肌灌注断层显像的半定量指标EG／EF、FG／EF和EF/AB可用于心肌梗死后左室重构的评价．