Development of 177Lu-EB-RGD molecular probe and its imaging and therapy in the patient-derived xenografts of non-small cell lung cancer / 中华核医学与分子影像杂志

Objective:To develop a novel α vβ 3-targeted theranostic agent 177Lu-Evans blue (EB)-Arg-Gly-Asp (RGD) and evaluate its value for SPECT imaging and targeted radionuclide therapy in the non-small cell lung cancer (NSCLC)-patient-derived xenografts (PDX). Methods:The α vβ 3-targeted molecule RGD was conjugated with the albumin binding moiety EB to obtain EB-RGD, and EB-RGD was further conjugated with the chelator 1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA) for 177Lu radiolabeling. NSCLC-PDX mice models ( n=68) were established. 177Lu-EB-RGD SPECT imaging, biodistribution study were performed in 28 PDX mice models after being injected with 177Lu-EB-RGD or 177Lu-RGD. Targeted radionuclide therapy were subsequently performed in NSCLC-PDX mice models, saline group (group A), 18.5 MBq 177Lu-RGD group (group B), 18.5 MBq 177Lu-EB-RGD group (group C), 29.6 MBq 177Lu-EB-RGD group (group D), n=10 in each group; tumor volumes of PDX mice models in each group were observed within 50 d. Differences between 2 groups were compared using independent-sample t test. Results:177Lu-EB-RGD was radiolabeled at a specific activity of (55±14) GBq/μmol, with a radiochemical yield of more than 95% and a radiochemical purity of more than 95%. Regarding the SPECT imaging, tumors in NSCLC-PDX mice were clearly observed from 4 to 96 h post-injection and the tumor to muscle ratio (T/M) reached 7.34±0.67, 14.63±3.82, 15.69±3.58 and 15.99±5.42 at 4, 24, 72, 96 h post-injection, respectively. Biodistribution study further confirmed the findings from SPECT imaging, and the tumor uptake of 177Lu-EB-RGD were markedly increased compared to 177Lu-RGD 4 h post-injection ((10.15±1.17) vs (3.30±1.47) percent injection dose per gram (%ID/g); t=18.60, P<0.05). Regarding targeted radiotherapy, the tumor volumes were quickly increased within 50 d after treatment in group A and B, while the tumor volumes were decreased in group C and D, until the tumors in group C and D disappeared at the 28th day after initial treatment with no sign of recurrence during the observation period. Conclusions:177Lu-EB-RGD can target α vβ 3-positive NSCLC-PDX with intense tumor to background ratio and strong tumor inhibition efficacy. The preclinical data suggests that 177Lu-EB-RGD may be an effective new treatment option for advanced NSCLC patients with resistance or ineffective results for targeted therapy.

Diagnostic value of a novel RGD-peptide based tracer 18F-Alfatide II for breast cancer / 中华核医学与分子影像杂志

Objective To investigate the biodistribution of 18F-Alfatide II in patients with breast diseases and to compare its uptake with 18F-fluorodeoxyglucose(FDG)uptake.Methods A total of 44 female patients(age:(50.7±8.0)years)with clinically suspected breast cancer from December 2015 to May 2017 were prospectively enrolled and underwent 18 F-Alfatide II and 18F-FDG PET/CT prior to treatment.By drawing regions of interest in normal organs and breast lesions,differences between 18F-Alfatide II uptake and l8F-FDG uptake were evaluated in all patients.Paired t test,two-sample t test and Wilcoxon rank sum test were used for data analysis.Results There were 53 breast lesions confirmed by histopathology in 44 patients.Among them,42 lesions were malignant and the others were benign.The uptake of 18F-Alfatide II was very low in the brain,vocal cords,lungs,blood pool and muscle.But the renal cortex and bladder had high 18F-Alfatide II accumulation.Different levels of 18F-Alfatide II uptake were found in other normal organs including normal breast tissue.There were differences(t values:2.04-41.65,all P<0.05)between 18F-Alfatide II and 18F-FDG maximum standardized uptake value(SUVmax)and mean standardized uptake value(SUVmean)in many normal organs except for the choroid plexus,salivary glands,liver,colon and normal breast tissue.The uptake of 18F-Alfatide II was significantly lower than 18F-FDG in breast cancer lesions(SUVmax:3.77±1.78 vs 7.37±4.48,SUVmean:2.25±0.98 vs 4.54±2,82;t values:4.89,4.82,both P< 0.05),but it was still higher in benign breast lesions(SUVmax:2.37±1.62,SUVmean:1.50±0.92;t val-ues:2.35,2.29,both P<0.05).Also,target/non-target(T/NT)of 18F-Alfatide II in breast cancer lesions was higher than that in benign breast lesions(5.32±3.08 vs 2.60±2.37;t = 2.72,P<0.05).Condusion The biodistribution of 18F-Alfatide II in patients is favorable and 18F-Alfatide II can be clinically used for breast cancer imaging.

Preclinical microPET imaging in tumor-bearing nude mice and pharmacokinetic studies of 18F-Alfatide II / 中华核医学与分子影像杂志

Objective To assess the imaging characteristics of 18F-Alfalide II in different tumorbearing mice and pharmacokinetics in Beagle dogs.Methods BALB/c nude mice(n-24)were used for subcutaneous tumor models(A549 and U87MG),orthotopic lung cancer models(A549)and orthotopic breast cancer models(MDA-MB-231)(n=6 in each group).18F-Alfatide II and 18F-fluorodeoxyglucose(FDG)microPET/CT images were compared in the 4 types of tumor-bearing nude mice models.18F-Alfatide II blocking experiment,biodistribution experiment and imaging studies in tumors of different growth cycles were performed in A549 subcutaneous tumor-bearing nude mice models.Pharmacokinetic experiments were carried out in Beagle dogs(n = 6)and CD-1 mice(n = 9).Two-sample t test was used to analyze the data.Results Compared with 18F-FDG,18F-Alfatide II microPET/CT images showed better imaging quality and contrast in subcutaneous A549,U87MG tumors and orthotopic A549(tumor/heart:4.50±1.17 vs 0.95±0.31;t = 4.125,P<0.01),orthotopic MDA-MB-231(tumor/muscle:6.60±1.53 vs 0.92±0.43;t = 3.984,P<0.01)transplantation nude mice models.18F-Alfatide II could specifically target A549 tumors,and the tumor uptake of 18F-Alfatide II was reduced by about 75% after pre-injection with cyclo(Arg-Gly-Asp-D-Tyr-Lys)(c(RGDyk)).18F-Alfatide II was rapidly cleared from the blood of Beagle dogs(T1/2 was(57.34±11.69)min).It was cleared in the form of prototype drug and(69.24±6.82)% of cumulative dose was excreted through the urine within 4 h after administration.Conclusions 18F-Alfatide II shows a higher target/non-target ratio than,18F-FDG in the imaging of A549,MDA-MB-231 and U87MG tumor-bearing nude mice models,which is more conducive to the diagnosis of tumor.18F-Alfatide II has excellent pharmacokinetic properties.

Factors for the synthesis of 18F-MyoZone and myocardial tomography in normal swine / 中华核医学与分子影像杂志

Objective To study the factors affecting the synthesis of 18F-MyoZone,and to evaluate its potential as a myocardial perfusion imaging (MPI) agent in normal Chinese mini-swine.Methods 18F-MyoZone was prepared by substituting the leaving group toluenesulfonyloxy (OTs) from the precursor compound with 18F-fluoride (18F-F-).The conditions affecting the labeling yield were studied by varying the amount of K2CO3 and precursor compound,18F-fluorination reaction time and temperature.PET was performed at 5,30,60 and 120 min post-injection on normal Chinese mini-swine.Results The doses of K2CO3 and precursor,the reaction time and the reaction temperature could affect the labeling yield of 18F-MyoZone,especially K2CO3.The optimized synthetic condition was 1.0 mg K2CO3,2.0 mg mpp2-OTs,20 min reaction time at 90 ℃.The total radio-synthesis time in this condition was 60 min.The uncorrected radiochemical yield was (24.0±5.1) ％.The radiochemical purity was ＞98％.PET imaging showed that 18F-MyoZone had high initial uptake (SUV=8.17± 1.83 at 5 min post-injection) and good retention (SUV =5.78±0.99 at 120 min post-injection) in the heart.The clearance of 18F-MyoZone from liver was very fast.The heart/liver ratios were 3.32,5.31,6.09 and 5.76 at 5,30,60 and 120 min post-injection,respectively.From 5 to 120 min post-injection,the outline of heart was clear and intact.There was almost no interference from the adjacent organs.The quality of PET images was highly satisfactory.Conelusions 18 F-MyoZone has the potential to be a good myocardial perfusion agent.The amount of K2CO3 used could significantly affect the labeling yield of 18F-MyoZone.

Synthesis and preliminary evaluation of 18F-AIF-NOTA-G-TMTP1 for PET imaging on nude mice bearing high-metastatic potential hepatoma cells / 中华核医学与分子影像杂志

Objective To synthesize 18F-AlF-NOTA-G-TMTP1 and evaluate its potential for PET imaging on nude mice bearing high-metastatic potential hepatoma cells.Methods NOTA-G-TMTP1 was synthesized by the standard Fmoc-solid phase synthetic protocols and radiolabeled with 18F using NOTA-AlF chelation method.The nude mice models bearing low-metastatic potential HCC97L and high-metastatic potential HCCLM3 xenografts were established separately.The tumor-targeting characteristics of 18F-AlF-NOTA-G-TMTP1 were assessed by microPET/CT and biodistribution assay.Results NOTA-G-TMTP1 was labeled with 18F in one step with (25±6)％ labeling yield (n=5).The radiochemical purity of 18F-AlF-NOTA-G-TMTP1 was more than 95％ with a specific activity more than 11.1 GBq/μmol.The octanol/water partition coefficient (logP) for 18F-AlF-NOTA-G-TMTP1 was-3.166±0.022.The tumor to muscle ratios were 1.8± 0.4 and 4.7±0.2 at 35 min post injection for HCC97L and HCCLM3,respectively.The uptake of 18F-AlF-NOTA-G-TMTP1 in HCCLM3 tumor was inhibited (61.4％) by unlabeled G-TMTP1.Conclusion 18F-AlF-NOTA-G-TMTP1 has been successfully synthesized.It shows specific uptake by tumor induced by the high-metastatic potential hepatoma cells.

A study of Qishen Tongluo Zengzhi decoction for intervention of cognitive impairment in patients with acute phase of ischemic stroke / 中国中西医结合急救杂志

ObjectiveTo observe the effect of Qishen Tongluo Zengzhi decoction on cognitive impairment in patients with acute phase of ischemic stroke.Methods A prospective randomized controlled trial was conducted, and 130 patients with acute phase of ischemic stroke and cognitive impairment accompanied by Qi deficiency and blood stasis and stagnationof phlegm-dampness syndrome admitted into the Neurology and Rehabilitation Departments of Rizhao Hospital of Traditional Chinese Medicine(TCM) Affiliated to Shandong University of TCM were randomly divided into treatment group and control group, 65 cases in each group. In the two groups, conventional internal treatment was given to all patients, and in the treatment group, additionally the Qishen Tongluo Zengzhi decoction was administered orally(composition: astragalus membranaceus 30 g, radix pseudostellariae 30 g, notoginseng 10 g,spatholobus stem 25 g, hirudo 3 g, pberetima 10 g, radix paeoniae rubra 12 g, Chinese angelica 12 g, peach kernel 10 g, carthamus tinctorious 10 g, achyranthes 12 g, radix rhapontici 10 g, rhizoma alismatis 6 g, Acorus gramineus Soland 9 g, polygala root 9 g, rhizoma cyperi 10 g, herba siegesbeckiae 15 g),one dose a day. While in the control group, oxiracetam 4.0 g intravenous drip was given, once a day. The whole course was 21 days in both groups. Before and after treatment, the cognitive function of all the patients in two groups was assessed by Mini-Mental State Examination(MMSE)and Montreal Cognitive Assessment(MoCA) scores, and incubation period and amplitude of P300 wave were recorded.Results Finally 62 cases were in treatment group and 63 cases in control group. Before treatment, the comparisons of the MMSE score, MoCA score and P300 latent period and amplitude between the two groups had no statistically significant differences(allP>0.05). After treatment in the two groups, the MMSE score, MoCA score and P300 wave amplitude were elevated, P300 latency period was shortened compared with those before treatment, and the changes were more prominent in treatment group〔score of MMSE: 25.33±2.32 vs. 21.68±2.29, score of MoCA(score): 26.61±3.06 vs. 22.40±2.93, P300 wave incubation period(ms): 349.62±20.01 vs. 371.87±19.63, P300 wave amplitude(μV): 8.70±2.92 vs. 5.72±2.33,allP<0.01〕.ConclusionQishen Tongluo Zengzhi decoctioncan effectively intervene cognitive impairment in patients with acute phase of ischemic stroke, and improve their cognitive function.

Cloning, molecular characterization and expression of acireductone dioxygenase (ARD) gene from Salvia miltiorrhiza / 中国中药杂志

<p><b>OBJECTIVE</b>To study the acireductone dioxygenase (designated as SmARD) gene of Salvia miltiorrhiza through bioinformatics and characterization of its tissue expression and response expression on stress in shoot.</p><p><b>METHOD</b>SmARD gene was obtained by sequencing cDNA library constructed by us. BLAST was used for alignment, ORF finder software was applied to find open reading frame, prosite was used to analyze the protein characterization. Semi-quantitative RT-PCR was used to detect the gene expression level.</p><p><b>RESULT</b>The full -length cDNA of SmRAD was 688 bp long with a 591 bp ORF (open reading frame) that putatively encoded a polypeptide of 196 amino acids; with a predicted molecular mass of 23.27 kDa. The deduced amino acid sequence of SmRAD of gene shared high homology with other known RADs. Semi-quantitative RT-PCR analysis indicated that SmRAD was constitutively expressed in roots, stems, flower and leaves of S. miltiorrhiza, with the high expression in roots. In addition, SmRAD expression level under different stress condition was also analyzed in root, including signaling components for plant defence responses, such as methyl jasmonate, salicylic acid and ABA, as well as drought, cold and salt abiotic stress. The expression of SmRAD was suppressed by water deficit treatment for 3 d, 150 mmol x L(-1) NaCl, 4 degrees C cold and 100 mmol x L(-1) ABA treatment for 1 d, but induced by 100 mmol x L(-1) MJ and 10 mmol x L(-1) ETH.</p><p><b>CONCLUSION</b>A novel SmARD gene was cloned from S. miltiorrhiza. This study will enable us to further understand the role of SmARD in the defense response under different abiotic stress and in synthesis of active cmpounds in S. miltiorrhiza at molecular level.</p>

Open reduction and internal fixation for Die-punch fracture of distal radius / 中国矫形外科杂志

[Objective]To discuss the surgical technique and clinical effects of open reduction and internal fixation with T style plate in the treatment of Die-punch fracture of distal radius.[Method]From August 2003 to October 2007,twenty-nine patients diagnosed as Die-punch fracture according to X-ray and computerized tomography were treated with open reduction through volar approach and internal fixation with T style plate and bone-grafts to support the articular surface of impacted fractures.There were 20 males and 9 females,with an average of 40.5 years(range 19-62 years).The fractures were caused by falling(n=23),traffic(n=4) and by athletics(n=2).There were 6 cases of open fracture.Eighteen cases were in the right wrist,and 11 in the left wrist.The displacement of articulatzons was from 1.5 to 3.5 mm.[Result]Twenty-six patients were followedup for 9 to 45 months,with an average of 28 months.Posteroanterior and lateral radiographs were taken periodically and fracture healing occurred all within 12 weeks.According to ADL standard and radiographs,21 cases were excellent(ADL scores:38-40),3 cases were good(scores:35-37).The excellent to good rate was 92.3%.No infection occurred.[Conclusion]Open reduction and internal fixation can help restore the smoothness of the articular surface and reduce the incidence of traumatic osteoarthritis.It is effective against the Die-punch fractures of distal radius.

Antimicrobial Activities of Traditional Chinese Materia Medica Extracts / 中华医院感染学杂志

OBJECTIVE To discuss the antimicrobial activity of extracts from 10 kinds of traditional Chinese materia(medica)(TCMM)in vitro.METHODS Extracts from 10 kinds of TCMM were prepared,and subjected to(bacteriostatic) tests in vitro by test tube continuously dilution in order to observe their minimal inhibitory(concentration)(MIC),to Staphylococcus(aureus),Escherichia coli,Pseudomonas aeruginosa and Candida albicans.RESULTS Among the 10 kinds of TCMM extracts,which did have antimicrobial activity against S.aureus,E.coli,P.aeruginosa and C.albicans in(different) degree;among them,the antimicrobial(activit)y of the extracts of Radix Paeoniae Rubra,Punica granatum,Schisandra chinensis and Coptis chinensis to resistant bacteria was the highest,the extracts of(Radix) Paeoniae Rubra showed strong inhibitory activities to the above 4 bacteria,especially to the resistant(organisms) with the MIC at 7.8,1.95,1.95 and 7.8mg/ml,(respectively).CONCLUSIONS The extracts of Radix Paeoniae Rubra,P.granatum,S.chinensis and C.chinensis have broad spectrum of antimicrobial activity to(resistant) bacteria.

Association between atherosclerotic brain infarction and human leucocyte antigen-Ⅱ gene of the Han Nationality in north China: Technique of sequent and specific primer polymerase chain reaction / 中国组织工程研究

BACKGROUND: The inheritance tendency and immunity imbalance of atherosclerotic brain infarction (ABI) have attracted the attention of many scholars at home and abroad, which suggests that immune mechanism plays a key role in ABI development. It has been found that ABI is related to polymorphism of human immune inheritance gene (HLA), particularly HLA-DR gene.OBJECTIVE: To investigate the correlation between HLA and ABI immune heredity.DESIGN: Single sample and single factor analysis.SETTING: Sino-Japanese Friendship Hospital of Jilin University; Neurological Department of the First Hospital of Harbin City.PARTICIPANTS: Totally 31 patients in experimental group and 30 healthy people in control group, whose three generations were of the Han Nationality in north China and who had no blood relation, were selected from Department of Neurology in First Hospital of Harbin between January 1998 and December 2000.METHODS: The experiment was conducted in the Blood Research Center of Department of Neurology in First Hospital of Harbin between June and December 2003.5 mL peripheral blood in both groups was taken to extract genome DNA with the previous method. Human HLA gene map had been described at gene bank in the Internet, in which mono-nucleic acid obtained from HLA of the sixth chromosome were retrieved. Each DR and DQ allele, sense primer and antisense primer were composed selectively. Amplification of DNA fragment was checked respectively and specifically. The primer was provided by Shanghai Gene Research Institute. PCR instrument (4 800) and its related reagents produced by PE Company (the US) were used. Fourfold table exact probability method was used to calculate the relative risk (RR) and P value.which was obviously higher than that of other sites (P ＜ 0.05). However,associated with the ABI susceptible gene of the Han Nationality in north China.