188Re-RGD-4CK的制备及其示踪动力学研究

目的:探讨188Re标记RGD-4CK的方法及其在健康家兔体内的示踪动力学特性.方法:采用预锡化法188Re直接标记RGD-4CK.纸层析测定标记多肽的标记率并计算比活度.测定9只家兔经耳缘静脉注射37 MBq 188Re-RGD-4CK后不同时相点血液中的放射性浓度,采用DAS软件评价标记多肽示踪动力学行为,以F值检验、AIC值、R2值并结合αvβ3a受体在正常机体表达实际.对血液放射性浓度-时间数据进行房室模型曲线拟合,根据拟合结果计算示踪动力学参数.结果:188Re-RGD-4CK的标记率大于97%,比活度为3.97±0.02TBq/mmol.188Re-RGD-4CK在健康家兔体内的放射浓度实测值符合以1/C2为权重系数拟合的二室模型理论计算值.结论:预锡化法188Re直接标记RGD-4CK制备方法简便,标记率高,无需分离纯化.188Re-RGD-4CK在健康家兔体内的示踪动力学符合以1/C2为权重系数拟合的二室模型.     

^131I标记Tyr-GX1在荷瘤裸鼠体内分布和显像研究

目的：设计、合成酪氨酸（Tyr）修饰的肿瘤血管靶向肽GX1,研究^131I标记短肽Tyr-GX1在荷人胃癌裸鼠体内的生物学分布与显像,探讨^131I-Tyr-GX1短肽作为肿瘤血管靶向诊治药物的可能性。方法：利用Iodogen碘标法对Tyr-GX1进行131I标记,检测其标记率和体内外稳定性;建立荷人胃癌裸鼠动物模型,尾静脉注射标记肽,分别进行体内生物学分布实验和肿瘤显像实验,结果用PASW Statistics18.0统计软件进行分析。结果：1）.纸层析法结果计算表明,^131I-Tyr-GX1肽的标记率和放化纯均达90%以上;24 h稳定性测试表明,^131I-Tyr-GX1在室温下存放以及与人血清、鼠血清、PBS等溶液混合,其标记率仍然都维持在90%左右,说明其具有良好的体内外稳定性;2）.荷瘤裸鼠体内生物学分布研究显示：标记肽在荷瘤裸鼠双肾放射性计数测量最高;其次是肝脏、肿瘤等组织;脑、骨、肌肉组织放射性计数含量较低,给药24 h时,肿瘤/肌肉（T/M）、肿瘤/血液（T/Bl）、肿瘤/脑组织（T/Br）的放射性比值分别是5.78、4.06和23.01;3）.体内SPECT显像结果显示：尾静脉注射^131I-Tyr-GX1肽后4 h肿瘤部位已开始显影,并随时间的延长,显像逐渐清楚,至18 h时,肿瘤显像最清晰。结论：应用Iodogen碘标法成功标记Tyr-GX1短肽;尾静脉注射^131I-Tyr-GX1后,肿瘤部位可以出现放射性浓聚,表明^131I-Tyr-GX1短肽可以靶向结合于肿瘤部位,有望成为新一种胃肠道肿瘤诊断与治疗的药物。     

放射性锆在模拟海水、底质及海洋生物系统中的动态

研究了放射性锆(^95Zr)在模拟海水、底质及海洋生物间的迁移、消长的动态．结果表明,海水中^95Zr的比活度在最初4h内下降了90％以上,此后呈缓慢递减趋势;底质中^95Zr的比活度至48h达到最大值,随后逐渐降低;半褶织纹螺螺壳和螺肉的放射性百分比分别为68．7％和31．30％;大弹涂鱼的鳃、鳍、内脏、头骨、皮、脊柱和肌肉等组织的放射性百分比分别为22．80％、12、64％、34．82％、10．31％、4．48％、11．55％和3．71％．半褶织纹螺对^95Zr的积累比大弹涂鱼更强、^95Zr在海水、底质、半褶织纹螺和大弹涂鱼中浓度的大小顺序是：底质>半褶织纹螺>海水>大弹涂鱼、各分室中^95Zr浓度(即比活度)随时间变化的动态可用指数函数表达．     

125 I标记白眉蝮蛇毒精氨酸酯酶代谢动力学研究

研究白眉蝮蛇(Agkistrodon halys ussuriensis)毒精氨酸酯酶的代谢动力学,为临床应用提供依据。125I标记的精氨酸酯酶对大鼠一侧颈静脉给药,不同时间从另一测颈静脉取血。5h后处死动物,取各组织、尿液、胆汁和粪便,对各样本的放射性进行测定并拟合时间-放射性关系曲线。代谢动力学拟合曲线符合一室模型,其中生物半衰期T1/2为55.9min,K值为0.0124min-1。125I标记的精氨酸酯酶在体内各组织广泛分布,但有血脑屏障存在,肝、肾和尿液中的放射性比其它组织要高很多,主要通过肝脏降解,肾脏排泄。     

^125I标记白眉蝮蛇毒精氨酸酯酶代谢动力学研究

研究白眉蝮蛇(Agkistrodon halys ussuriensis)毒精氨酸酯酶的代谢动力学,为临床应用提供依据。125I标记的精氨酸酯酶对大鼠一侧颈静脉给药,不同时间从另一测颈静脉取血。5h后处死动物,取各组织、尿液、胆汁和粪便,对各样本的放射性进行测定并拟合时间-放射性关系曲线。代谢动力学拟合曲线符合一室模型,其中生物半衰期T1/2为55.9min,K值为0.0124min-1。125I标记的精氨酸酯酶在体内各组织广泛分布,但有血脑屏障存在,肝、肾和尿液中的放射性比其它组织要高很多,主要通过肝脏降解,肾脏排泄。     

铼-188标记生物分子在肿瘤治疗中的应用

目的：研究铼．188标记生物分子在肿瘤治疗中的应用。方法：选取小白鼠作为实验的研究对象,将荷瘤鼠的肉瘤切成小块接种到小白鼠身上,达到试验条件时使用,即将没有明显差异的小白鼠16只随机分为4组,每组4只,注射含有铼一188的药物后分别在不同的时间将其处死,之后取出重要器官进行测量分析,进而得出铼一188的应用效果。结果：瘤内注射的要去在不同时间放射性在瘤内的保持率分别为（90．5±7．7）D％（1h）,（92．2±8．6）D％（24h）,（88．3±10．9）D％（48h）和（91．5±7．6）D％（72h）,在荷瘤鼠内注入生理盐水、非放硫化铼和188Re．硫化铼混悬液,肿瘤质量分别为2885．3±1241.3、2839．9±1965．2和98．4±45．5mg。188Re-硫化铼混悬液在生理盐水、磷酸盐缓冲液和小牛血清中均可稳定72h,而且188W-188Re发生器的应用还可以降低188Re-硫化铼混悬液的价格。随着处死时间的延迟,小鼠肿瘤质量和体积逐渐减小,相临两组比较,后组测定值均明显小于前组,数据经统计学比较具有显著差异（P〈0．05）。结论：188Re-硫化铼混悬液是一种适宜的肿瘤治疗剂。     

重组人甲状旁腺素（1-34）在大鼠体内的组织分布和排泄

目的：研究重组人甲状旁腺素（1-34）[rhPTH（1—34）]在大鼠体内的组织分布和排泄情况,为进一步的临床实验提供参考。方法：用^125I-同位素示踪法结合TCA酸沉淀法测定各主要器官组织的总放射性浓度和酸沉淀部分放射性浓度,获得rhPTH（1-34）的尿粪排泄和胆汁排泄数据。结果：各主要器官组织的总放射性浓度排序由高到低依次为：尿、肾、膀胱、肠内容物、肌肉、血清、肾上腺、空肠、肝、肺脏、卵巢、肠淋巴结、脾、胸腺、心脏、脂肪、睾丸和脑;大鼠皮下注射。^125I-rhPTH（1-34）后,骨骼组织中放射性分布低于血浆,但消除缓慢,血浆浓度4h较15min降低了78％,而骨骼浓度多数仅降低了50％以下;注射后72h,尿、粪分别排出注入放射性量的73．6％±10．9％和3．2％±1．3％,尿、粪合计排出注入放射性量的76．8％±11,4％;注射后12h,胆汁中累积排出注入放射性的6．64％±1．04％。经分子筛排阻HPLC证实,^125I-rhPTH（1-34）不与大鼠的血浆蛋白发生结合。结论：rhPTH（1-34）在泌尿系统中的分布较高,在脂肪和脑中最低,提示药物不易透过血脑屏障;就全身放射性分布而言,在骨骼中分布较高,提示药物具有一定的靶向性;rhPTH（1-34）主要经尿的形式排泄。     

~(125)Ⅰ—标记尖吻蝮蛇出血毒素Ⅰ在家兔体内的药代动力学研究

用~(125)Ⅰ标记从尖吻蝮蛇(Agkistrodon acutus)毒中分离出的出血毒素(Ⅰ Aa-HI),得到Ⅰ~(125)Ⅰ—AaHI。静脉注射~(125)Ⅰ—AaHI到家兔体内,对~(125)Ⅰ—AaHI在动物体内的分布和药物代谢动力学进行研究。注射~(125)Ⅰ—AaHI 5小时后将家兔杀死,测定各组织的放射性强度。结果表明有血脑屏障存在。~(125)Ⅰ—AaHI代谢的大量产物由肾通过尿排出。对于药物代谢动力学,计算机模似结果为一室模型,其中生物半衰期T_(1/2)为55.9分钟,K值为0.0124分钟。我们认为在动物体内可能有AaHI相关的结合位点或受体存在。     

用BIAcore系统研究西妥昔单克隆抗体与表皮生长因子受体的结合

目的：应用基于表面等离子体共振技术的BIAcore3000系统研究国产西妥昔单克隆抗体（cetuximab）C225与可溶性重组人表皮生长因子受体（EGFR）的结合能力,并与国外已上市的西妥昔单抗Erbitux相比较。方法：在CM5传感器芯片上设置2个通道,一个氨基偶联重组人EGFR作为检测通道,另一个不固定EGFR作为空白参比通道;以HBS溶液作为工作液,流速为10μL／min;活化与封闭芯片;再以10μL／min的流速分别以梯度浓度进样C225和Erbitux,每个浓度级别检测2次;获得结合动态图谱,拟合处理后用软件模块进行参数计算。结果：C225与可溶性重组人EGFR的结合动力学常数K^为4．00×10^8L／mol,KD为2．50×10^-9mol／L;而Erbitux与可溶性重组人EGFR的结合动力学常数KA为4．25×10^8L／mol,KD为2．35×10^-9mol／L。结论：在与可溶性重组人EGFR的结合能力上,C225与Erbitux有相似的结合动力学特性。     

表达CD19^＋CD56^-的多发性骨髓瘤浆细胞免疫表型的鉴别

目的：研究多发性骨髓瘤患者浆细胞和对照组正常浆细胞免疫表型,有效地识别表达CD19^＋CD56^-的多发性骨髓瘤恶性浆细胞。方法：采用四色流式细胞仪（BD,FACSCalibur）检测44例MM患者浆细胞以及25例健康骨髓捐献者正常浆细胞膜上的抗原表达。采用CellQuest软件分析结果。细胞膜表面抗原表达率大于20％定义为表达阳性。阳性率为表达阳性的患者及健康对照者所占百分比。结果：44例MM患者浆细胞免疫表型表达频率为：CD^138＋;97．72％（43／44）、CD38^＋：100％（44／44）、CD56^＋：63．64％（28／44）、CD19：84．09％（37／44）、CD200^＋：77．27％（34／44）、CD28^＋：38．64％（17／44）;25例对照组正常浆细胞免疫表型表达频率为：CD138^＋：100％（25／25）、CD38^＋：100％（25／25）、CD56^＋-：100％（25／25）、CD19^＋：96％（24／25）、CD200^＋：0％（0／25）、CD28^＋：0％（0／25）。在44例MM患者中,9％（4／44）患者表达CD19^＋CD56^-,利用CD200检测4例表达CD19^＋CD56^-的患者,有3例患者伴有CD200阳性表达,可与正常浆细胞鉴别。结论：CD200有利于鉴别表达CD19＋CD56-MM恶性浆细胞与正常浆细胞。     

重组葡激酶（r—Sak）在家兔体内的药代动力学研究

家兔静脉注射重组葡激酶(r-Sak)后,经用二室模型(权重系数1/C)计算。结果表明,该药分布相较短,t1/2α约为1.11±0.72 min,消除也快,t1/2β约为8.64±0.21min,但在胆汁及尿液中未测出r-Sak的生物活性。     

90Y labeled phosphorodiamidate morpholino oligomer for pretargeting radiotherapy

While (188)Re has been used successfully in mice for tumor radiotherapy by MORF/cMORF pretargeting, previous radiolabeling of the amine-derivatized cMORF with (90)Y, a longer physical half-life nuclide, was not very successful. After developing a method involving a prepurification heating step during conjugation that increases labeling efficiency and label stability, the biodistribution of (90)Y-DOTA-Bn-SCN-cMORF ((90)Y-DOTA-cMORF) was measured in normal mice and in MORF-CC49 pretargeted mice that bear LS174T tumors. Absorbed radiation doses were then estimated and compared to those estimated for (188)Re. The pharmacokinetics of the (90)Y-DOTA-cMORF in normal mice and in the pretargeted nude mice was similar to that observed previously with (99m)Tc- and (188)Re-MAG(3)-cMORFs. While the (90)Y-DOTA-cMORF cleared rapidly from normal tissues, tumor clearance was very slow and tumor radioactivity accumulation was constant for at least 7 days such that the tumor/blood (T/B) ratio increased linearly from 6 to 25 over this period. Therefore, by extrapolation, normal tissue toxicities following administration of therapeutic doses of (90)Y may be comparable to that observed for (188)Re in which the T/B increased from 5 to 20. In conclusion, radiolabeling of DOTA-cMORF with (90)Y was improved by introducing a prepurification heating step during conjugation. The (90)Y-DOTA-cMORF provided a similar T/B ratio and biodistribution to that of (188)Re-MAG(3)-cMORF and was retained well in the tumor pretargeted with MORF-CC49. Because of the longer physical half-life, the T/NT absorbed radiation dose ratios were improved in most organs and especially in blood.     

Novel and efficient preparation of precursor [188Re(OH2)3(CO)3]+ for the labeling of biomolecules

A novel and efficient method for preparing 188Re(I) tricarbonyl precursor [188Re(OH2)3(CO)3]+ has been developed by reacting [188Re]perrhenate with Schibli's kit in the presence of borohydride exchange resin (BER) as a reducing agent and an anion scavenger. The precursor was produced in more than 97% yield by reacting a solution of tetrahydroborate exchange resin (BER, 3 mg), borane-ammonia (BH3.NH3, 3 mg), and potassium boranocarbonate (K2[H3BCO2], 3 mg) in 0.9% saline with a solution of sodium perrhenate (Na188ReO4) with up to 50 MBq and concentrated phosphoric acid (85%, 7 microL) at 60 degrees C for 15 min. HPLC and TLC revealed 0% unreacted [188Re]perrhenate ion and <3% of colloidal 188ReO2. Since the precursor is produced with high radiochemical purity and labeling efficiency under the milder conditions than those required for the conventional reducing agents, the latter can be replaced.     

On the role of mi-cK and VDAC in mitochondrial function of heart muscle cells

A two-compartment kinetic model was used to describe reconstituted systems in which mitochondria compete with pyruvate kinase for kinase-generated ADP. The modelling suggests that cytosolic CK deficiency results in a 50% increase in outer mitochondrial membrane permeability.     

Suppression of murine collagen-induced arthritis by targeted apoptosis of synovial neovasculature

Because angiogenesis plays a major role in the perpetuation of inflammatory arthritis, we explored a method for selectively targeting and destroying new synovial blood vessels. Mice with collagen-induced arthritis were injected intravenously with phage expressing an RGD motif. In addition, the RGD peptide (RGD-4C) was covalently linked to a proapoptotic heptapeptide dimer, _D(KLAKLAK)₂, and was systemically administered to mice with collagen-induced arthritis. A phage displaying an RGD-containing cyclic peptide (RGD-4C) that binds selectively to the αvβ3 and αvβ5 integrins accumulated in inflamed synovium but not in normal synovium. Homing of RGD-4C phage to inflamed synovium was inhibited by co-administration of soluble RGD-4C. Intravenous injections of the RGD-4C–_D(KLAKLAK)₂ chimeric peptide significantly decreased clinical arthritis and increased apoptosis of synovial blood vessels, whereas treatment with vehicle or uncoupled mixture of the RGD-4C and the untargeted proapoptotic peptide had no effect. Targeted apoptosis of synovial neovasculature can induce apoptosis and suppress clinical arthritis. This form of therapy has potential utility in the treatment of inflammatory arthritis.     

HPLC法同时测定人参皂苷Rb1、Rc、Rd、Rg3、CK和Rh2

目的:建立高效液相色谱法同时测定人参皂苷Rb1、Rc、Rd、Rg3、CK和Rh2的方法.方法:采用ODSC18(4.6 mm×150 mm)色谱柱,流动相乙腈-0.05%磷酸水,梯度洗脱,流速1 Ml/min,检测波长203 nm,柱温35 ℃.结果:人参皂苷Rb1、Rc、Rd、Rg3、CK和Rh2分离效果良好,线性关...     

The effect of locally delivered c-myc antisense oligonucleotides combined with intravascular brachytherapy of 188Re liquid-filled balloon therapy on vascular smooth muscle cell proliferation in rabbit iliac arteries after injury with a balloon catheter.

Poststenting restenosis is a significant clinical problem that involves vascular smooth muscle cell (VSMC) proliferation. We primarily investigated the effect of c-myc antisense oligodeoxynucleotides (ASODNs) combined with 188Re radiation therapy on VSMC proliferation in rabbit common iliac arteries injured by the porous balloon catheter to explore the therapeutic potential of the combined therapy for the prevention of restenosis. The iliac arteries in rabbits were injured with balloon catheters, and radiation therapy was carried out with a 2.5 mm balloon catheter filled with 188Re (8 or 15 Gy), and ASODNs (300 microg) were applied to the adventitia introduced using a pluronic gel releasing system and a lipofectin delivery system. After 3 weeks, the animals were killed and defined segments of arteries were sectioned. The histological sections were stained using alpha-actin immunohistochemistry staining. The positive alpha-actin ratios were calculated and analyzed statistically among groups. In contrast to the rate of alpha-actin positive cell staining in the control group, the rate of alpha-actin positive cell staining did not decrease (P > 0.05) in the 188Re-irradiated group (8 Gy). However, in the ASODN-treated group, the 188Re-irradiated group (15 Gy), and the combined ASODN - 188Reirradiated groups (8 or 15 Gy), the ratios had markedly decreased (P < 0.01). The effect of the combined group (ASODNs + 188Re (15 Gy)) provided the lowest level of alpha-actin positive cell staining (P < 0.01). The ASODNs (300 microg) effectively decreased VSMC poliferation. The effect of the 188Re radiation on the VSMCs depended on the dosage. The ASODNs (300 microg) and combined 188Re irradiation effectively lowered VSMC proliferation, and the effect was better than that achieved with any other treatment.     

TF-1细胞增殖法测定神经生长因子生物学活性的建立与应用

目的：建立并优化用于检测神经生长因子（NGF）生物学活性的TF-1细胞增殖法,并应用于重组人NGF和鼠NGF制品的活性检测。方法：将梯度稀释的人NGF国际参考品与TF-1细胞相作用,通过MTT法测定细胞增殖情况,建立测定NGF活性的TF-1细胞增殖法;从粒细胞巨噬细胞集落刺激因子（CM-CSF）残留、NGF加样稀释率、TF-1细胞接种浓度、培养时间等多个环节对实验条件进行优化;将建立的TF-1细胞增殖法应用于重组人NGF和鼠NGF的活性检测。结果：建立了用于NGF活性检测的TF-1细胞增殖法;实验条件优化结果表明,在无GM-CSF残留的情况下,将稀释至1／4的NGF样品与4×10^5／mL的细胞相互作用,培养48h,可以得到更为典型的的四参数曲线;利用条件优化后建立的检测方法对重组入NGF和鼠NGF各2批样品分别进行4次测定,结果平均值分别为10．01×10^5、10．81×10^5和3．55×10^5、3．30×10^5U／mg,变异系数分别为7．5％、7．2％和7．2％、9．1％,检测结果稳定均一,表明检测方法具有很好的重复性。结论：建立并优化了用于NGF活性检测的TF-1细胞增殖法,该方法操作简便、定量准确、重复性好、稳定可靠,可有效应用于人NGF和鼠NGF制品的活性检测。     

Leakage of a liquid 188Re-filled balloon system during intracoronary brachytherapy. A case report

We are reporting the first case of an accidental radioactive 188Re leakage of a liquid-filled balloon system. Different analytical methods estimated that approximately 4 mCi 188Re were released. The radiation burden was reduced considerably by the combined therapy with perchlorate and forced volume diuresis. Estimated exposures to all organs were very low with 1.8 rad. A total body nuclear scintigraphy demonstrated uniform 188Re distribution, without specific organ concentration.