^131 I—Rituximab对B细胞淋巴瘤生物学效应的实验研究

目的：研究^131 I标记的Rituximab对B细胞淋巴瘤细胞的生物学效应以及对荷人Raji细胞移植瘤裸鼠的放射免疫治疗效果,为放射免疫导向治疗提供实验依据。方法：体外培养人B细胞淋巴瘤细胞系Raji细胞,裸鼠皮下接种Raji细胞成瘤,IODO-GEN法将^131 I标记于Rituximab,将细胞分为^131 I—Rituximab组、单纯抗体组、单纯核素组及空白对照组4组,流式细胞仪检测各组Raji细胞凋亡和细胞周期;取30只成瘤裸鼠随机分成高、中、低剂量治疗组及单纯抗体组、单纯核素组、空白对照组6组,进行裸鼠的放射免疫治疗研究。每周测量裸鼠荷瘤大小1次,4周后处死裸鼠,取瘤称重,常规病理分析。结果：^131 I．Rituximab组凋亡率高于其他各组;^131 I—Rituximab组细胞周期发生变化,大部分被阻滞在G2期。裸鼠各治疗组与对照组比较,肿瘤生长减慢,肿瘤生长抑制率具有剂量时间依赖性,组织病理学检测显示治疗有效。结论：^131 I—Rituximab能够诱导Raji细胞凋亡并调控细胞周期,而且可特异性地定位于肿瘤组织,发挥放射免疫导向治疗作用,具有潜在的临床应用价值。     

~(131)I-Rituximab对B细胞淋巴瘤的放射免疫导向治疗的实验研究

目的:探讨放射性核素131I标记Rituximab及131I-Rituximab对B细胞淋巴瘤细胞进行放射免疫导向治疗的可行性。方法:采用氯胺-T法制备131I-Rituximab,采用CCK-8法观察131I-Rituximab对Raji细胞的抑制作用,对照组为131I、Rituximab组。结果:131I-Rituximab标记率在92%以上,放化学纯度达98%以上。CCK-8法表明131I-Rituximab对Raji细胞的抑瘤率与131I组和Rituximab组的抑瘤率有统计学差异(P<0.01)。结论:提示131I-Rituximab对人B细胞淋巴瘤的放射免疫导向治疗具有可行性。     

抗乙型肝炎表面抗原Fab片段联合抗细胞核单抗为载体的肝癌放射免疫治疗实验研究

目的 探讨核素标记人源化抗(乙型肝炎)乙肝表面抗原Fab片段(抗HBsAg Fab)联合核素标记抗细胞核单克隆抗体(chTNT)瘤内注射治疗荷人肝癌移植瘤裸鼠的可行性和优越性,为临床应用核素标记多种不同性质的单克隆抗体进行放射免疫治疗肝癌提供实验依据.方法 荷瘤裸鼠分为五组,分别瘤内注射131I-抗HBsAg Fab、131I-chTNT、131I-抗HBsAg Fab联合131I-chTNT、131I-无关抗体(131HgG)及PBS.治疗后行SPECT显像观察标记抗体在肿瘤内的浓聚,并澍量肿瘤大小变化,计算肿瘤抑制率.结果 研究发现2种标记抗体联合应用组的肿瘤抑制率为73.09%,呀显高于131I-抗HBsAgFab组的47.8%和131I-chTNT组的54.26%.联合应用组标记抗体在肿瘤组织/非肿瘤组织内的放射活度比值(T/NT值)大于单独应用组.结论 131I-抗HBsAg Fab联合131I-chTNT瘤内注射放射免疫治疗可增加标记抗体在肿瘤组织内的浓聚.并能提高放射免疫治疗效果.     

瘤内注射核素标记混合抗体治疗肝癌的实验研究

目的 为临床应用混合抗体进行瘤内注射放射免疫治疗提供实验依据。方法 在裸鼠荷人肝癌2.2.15移植瘤生长至1 cm时,瘤内分别或同时注射131I标记的抗癌胚蛋白（AFP）单克隆抗体（131I -AFP-IT）和131I标记抗乙型肝炎表面抗原单克隆抗体S102（131I- S102-IT）,治疗后行SPECT显像观察标记抗体在肿瘤内的浓聚,并进行疗效观察。 结果 发现两种标记抗体联合治疗组肿瘤抑制率为76.5%,明显高于131I -AFP-IT组的53.1%及131I- S102-IT组的47.3%,抗体在肿瘤内的浓聚也明显高于单独应用组。结论 多种抗体混合应用瘤内注射放射免疫治疗可增加标记抗体在肿瘤内的浓聚,并能提高放射免疫治疗的疗效。     

131Ⅰ-Rituximab对B细胞淋巴瘤细胞杀伤效应的实验研究

目的 研究131Ⅰ标记的Rituximab对CD20表达阳性的B细胞淋巴瘤细胞的特异性杀伤作用,为放射免疫导向治疗提供实验依据.方法 应用IODO-GEN法对Rituximab进行131Ⅰ标记,以MTT比色法测定131Ⅰ-Rituximab、131Ⅰ和Rituximab对Raji细胞的剂量效应曲线,并根据剂量效应曲线选取合适的剂量,以131Ⅰ-Rimximab、131Ⅰ及Rituximab作用于CD20阳性的Ramos RA-1细胞、Raji细胞以及CD20阴性的Molt-4细胞,根据细胞存活率的变化,评价131Ⅰ-Rituximab、131Ⅰ及Rituximab对上述细胞的杀伤作用.Gimesa染色观察核分裂指数(MI值)等指标评价131Ⅰ-Rituximab对Raji细胞的抗肿瘤活性.结果 131Ⅰ-Rituximab对Raji细胞的杀伤作用呈剂量依赖性;选取60μCi/ml作为疗效实验的作用剂量,131Ⅰ-Rituximab组的细胞抑制率显著高于Rituximab组(P<0.05).Raji细胞在131Ⅰ-Rimximab、131Ⅰ、Rituximab作用96h后的细胞抑制率,与同等条件下的Ramos(RA-1)细胞比较,无显著性差异(P0.05);与同等条件下的Molt-4细胞比较,均显著增高(P<0.05).④131Ⅰ-Rituximab的MI值最低,与其他各组比较有显著性差异(P<0.001).结论 131Ⅰ-Rituximab可特异性杀伤CD20表达阳性的肿瘤细胞.为放射免疫治疗CD20阳性的B细胞淋巴瘤提供可能性.     

核素标记混合抗体瘤内注射治疗大肠癌的实验研究

目的:为临床应用混合抗体进行放射免疫治疗提供实验依据。方法:在裸鼠荷人大肠癌Lovo移植瘤生长至1cm时,瘤内分别或同时注射~(131)I标记的抗CL_3单克隆抗体(~(131)I-CL_3)和~(131)I标记抗FERR单克隆抗体T_9(~(131)I-T_9),治疗后行SPECT显像观察标记抗体在肿瘤内的浓聚,并进行疗效观察。结果:我们发现两种抗体混合治疗组疗效明显好于单独治疗组,抗体在肿瘤内的浓聚也大于单独应用。结论:多种抗体混合应用瘤内注射放射免疫治疗可增加标记抗体在肿瘤内的浓聚,并能提高放射免疫治疗的疗效。     

131I-rituximab对B细胞淋巴瘤细胞生物学效应的实验研究

目的 研究^131Ⅰ标记的rituximab对CD20高表达的B细胞淋巴瘤细胞的生物学效应,为放射免疫导向治疗提供实验依据。方法 IODO—GEN法将^131Ⅰ标记于抗CD20单抗rituximab,用AnnexinV—FITC／PI双染法检测^131Ⅰ-rituximab对Raji细胞的诱导凋亡作用,PI染色法检测细胞周期分布。结果 AnnexinV—FITC／PI双染法检测凋亡率：^131Ⅰ-rituximab组凋亡率为51．99％,^131Ⅰ组为42．71％．rituximab组为29．42％,对照组为26．17％。对照组和rituximab组凋亡率明显低于^131Ⅰ组和^131Ⅰ—rituximab组（P〈0．05）。PI染色法对比各组的凋亡率（亚二倍体峰）：^131ⅠI-rituximab组细胞凋亡率为4．32％,^131Ⅰ组为1．47％,rituximab组为1．39％,对照组仅0．37％,^131Ⅰ-rituximab组凋亡率明显高于其他各组（P〈0．05）。^131Ⅰ-rituximab组Raji细胞周期发生变化,细胞大部分被阻滞于G1／G2期。结论 ^131Ⅰ-rituximab能够调控Raji细胞的细胞周期并诱导其凋亡,从而抑制Raji细胞增殖。     

活体成像分析异硫氰酸荧光素标记Rituximab在荷淋巴瘤裸鼠体内的生物分布

目的 应用活体动物体内光学成像系统观察抗体药物Rituximab在荷淋巴瘤裸鼠体内的生物分布.方法 制备FITC标记的Rituximab(FITC-Rituximab),用激光扫描共聚焦显微镜和流式细胞仪体外分析FITC-Rituximab与人淋巴瘤Raji细胞的亲和力;建立裸鼠淋巴瘤皮下移植瘤模型,应用活体动物体内光学成像系统观察FITC-Rituximab在荷瘤小鼠体内的分布.结果 流式细胞仪和激光扫描共聚焦显微镜检测证明FITC-Rituximab与淋巴瘤Raji细胞有较好的亲合活性,主要结合在细胞膜表面.体内活体动物光学成像分析表明,FITC-Rituximab在1 h内即可特异性地在肿瘤部位富集,3～4 h即可进入肿瘤组织内部并达到最大浓度富集,8～10 h后在肿瘤组织仍可观察到FITCRituximab的特异性的富集,而在其他组织器官没有可见的荧光存在;双侧皮下移植瘤模型再次证明FITC-Rituximab具有对CD20抗原过表达的淋巴瘤特异性结合能力.结论 动物活体成像系统能够准确地监测FITC标记Rituximab在荷瘤裸鼠体内的动态分布情况,该技术对实时分析抗体药物在荷瘤小鼠体内的靶向效果具有参考价值和指导意义.     

131I-antiAFP导向载药纳米粒对肝癌移植瘤的抑制作用

目的 探讨131 I-antiAFP McAb-DOX-BSA-NP对肝癌移植瘤的抑制和杀伤作用.方法 制备131I-antiAFP McAb-DOX-BSA-NP;培养人肝癌细胞株BEL-7402及建立荷瘤裸鼠模型;将成瘤裸鼠模型按随机数字表法分为5组,每组6只.A组:空白对照组(生理盐水);B组:单纯131I核素治疗组;C组:131 I-antiAFP McAb治疗组;D组:131 I-antiAFP McAb-DOX-BSA-NP治疗组;E组:antiAFP McAb-DOX-BSA-NP治疗组.裸鼠瘤周注射治疗,DOX剂量为40 μg/只,131I活度0.2 mCi/只(1 mCi =3.7×107Bq),注射体积为0.2 mL/只.于4、9、14、20 d测量并记录各组裸鼠瘤体体积.同时分别在24、48、96、168 h对放射治疗组(B、C、D组)裸鼠进行SPECT检查.结果 标记产物呈澄清溶液;pH值7.4;放射性化学纯度＞99％,活度浓度3.91 mCi/mL;在0.2 mol/L PB缓冲液中1、2、4h放射性化学纯度分别为97.17％、97.10％、95.22％,稳定性良好.各组裸鼠治疗后肿瘤体积变化及生长抑制率比较:B、C、D、E治疗组较空白对照组肿瘤体积明显减小(P＜0.05);与B、C、E组比较,D组肿瘤生长抑制率最强,差异有统计学意义(P＜0.05).SPECT检查结果显示:24 h时,B、C、D3组瘤区放射性计数值差异无统计学意义(P ＞0.05);48、96、168 h时,131 I-antiAFP McAb-DOX-BSA-NP治疗组(D组)及131 I-antiAFP McAb治疗组(C组)放射性计数值均高于131I治疗组(B组),差异均具有统计学意义(P＜0.05),且随时间延长其差异性越来越显著.结论 单克隆抗体免疫治疗、放射性131I内照射治疗及载药纳米粒缓释化疗联合运用对肝癌移植瘤有显著抗癌效果.     

~(131)I-血管抑素联合PcDNA-sTRAIL在荷瘤小鼠模型肿瘤靶向治疗的研究

目的构建PcDNA-sTRAIL真核表达载体并进行扩增、纯化,采用131I标记血管抑素(AS)并完成了PcDNA-sTRAIL与131I-AS在荷瘤裸鼠体内的联合抑瘤试验。方法;通过PCR扩增和定向克隆技术成功构建PcDNA-sTRAIL真核表达载体,采用131I标记血管抑素,标记率达85%且体外稳定性好,成功构建了荷瘤裸鼠模型,并进行了荷瘤小鼠联合治疗后SPECT显像和病理结果,完成PcDNA-sTRAIL与131I-AS在荷瘤裸鼠体内的联合抑瘤试验。结果:荷瘤小鼠PcD-NA-sTRAIL与131I-AS联合用药治疗期间肿瘤平均体积较其他单独治疗组增长缓慢。结论:本研究将抑制肿瘤血管生成、促肿瘤细胞凋亡和放射性核素的内照射三重作用相结合并用于抗肿瘤治疗,期望不仅能充分发挥三者的优势,又能为核素内照射与免疫治疗的结合提供有力的实验证据。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.