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Effects of lentiviral vector-mediated RNA interference of hypoxia inducible factor-1α on the expression of related genes in human pancreatic cancer Patu8988 cells

Guanglei FAN ; Minbin DENG ; Yiwei WU ; Qixian YANG ; Bin ZHANG ; Longbao XU

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(5):377-380. doi:10.3760/cma.j.issn.2095-2848.2013.05.015

Objective To investigate the inhibitory effect of the lentiviral vector (LV)-mediated RNA interference (RNAi) targeting HIF-1α on the expression of HIF-1α and Glut-1 in human pancreatic cancer Patu8988 cells.Methods The RNAi targeting HIF-1α was combined to LV,and transfected into Patu8988 cells.The Patu8988 cells transfected with the empty vector and exposed to 0.5％ O2 for 4 h served as hypoxia negative control,the Patu8988 cells not transfected with vector and exposed to 0.5％ O2 for 4 h as hypoxia blank control,and the Patu8988 cells transfected with LV-RNAi-HIF-1α and exposed to 0.5％O2 for 4 h as experimental group.The expression of HIF-1α was measured by RT-PCR and Western blot respectively.The expression of Glut-1 was measured by RT-PCR.Each group was compared according to oneway analysis of variance and two-sample t test.Results After transfection with LV-RNAi-HIF-1α,HIF-1α mRNA expression decreased by 65.1％ (0.209/0.321) and 80.6％ (0.791/0.982) (t=10.52,15.24,both P＜0.05) under normoxia and hypoxia conditions,meanwhile with the empty vector,HIF-1α mRNA expression decreased by 0.6％ (0.002/0.321) and 7.2％ (0.071/0.982) (t =5.26,7.38,both P＜0.05).Under hypoxia conditions,the protein of HIF-1α in experimental group cells (0.159±0.010) was down-regulated obviously compared to the negative control group (0.745± 0.012) and the blank control group (0.711 ± 0.023)(F=35.52,t =6.72,10.56,all P＜0.05).The expression of Glut-1 mRNA in experimental group cells (0.040±0.003) decreased obviously compared to the negative control group (0.054±0.003) and blank control group (0.062±0.004) (F=35.28,t=5.94,8.55,all P＜0.01).Conclusion Gene silencing of HIF-1α using LV-mediated RNAi can inhibit the expression of HIF-1α and decrease the expression of Glut-1mRNA in Patu8988 cells.

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99Tcm-MDP imaging for the diagnosis of joint infection after total hip arthroplasty

Yuan LI ; Qian WANG ; Minggang YUE

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(4):267-270. doi:10.3760/cma.j.issn.2095-2848.2013.04.008

Objective To investigate the value of 99Tcm-MDP imaging for the differential diagnosis between infection and aseptic loosening after total hip arthroplasty.Methods During February 2008 to August 2011,74 patients (32 males,42 females,average age (64.3±11.2) years) with hip pain after arthroplasty underwent 3-phase (blood flow,blood pool and bone phases) 99Tcm-MDP imaging.All patients had measurements of serum C reactive protein (CRP) and erythrocyte sedimentation rate (ESR).Joint infection was defined as either increased peri-prosthetic soft tissue activity during perfusion and blood pool phases or increased peri-prosthetic bone activity during uptake phase,or positive in all 3 phases.Aseptic loosening was defined as having negative 99Tcm-MDP in all 3 phases.Clinical diagnosis was chosen as the gold standard.The blood flow-pool imaging was compared with the serum examinations.x2 test was used for statistical analysis.Results There were 74 symptomatic joints in 74 patients,including 24 joint infections and 50 aseptic loosening.For the detection of peri-prosthetic infection,combined perfusion-blood-pool phase was more accurate than bone uptake phase (90.5％ (67/74) vs 55.4％ (41/74) ;x2=23.159,P＜0.001),with the sensitivity of 91.7％ (22/24)vs 70.8％ (17/24) and specificity of 90.0％ (45/50) vs 48.0％ (24/50),respectively.The blood flow-pool imaging was also more accurate than CRP (73.0％ (54/74) ;x2 =7.656,P＜0.05) and ESR (71.6％ (53/74) ; x2 =8.633,P＜0.05),respectively.Conclusion 99Tcm-MDP perfusion/blood flow-pool imaging is an accurate modality for differentiating peri-prosthetic infection from aseptic loosening in patients with hip pain after arthroplasty.

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Evaluating the effects of an angiogenesis inhibitor of endostar with a novel RGD dimer probe labeled by 99Tcm

Yonghong SUN ; Xinhui SU ; Bing JIA ; Fan WANG ; Guibing CHEN ; Hua WU

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(4):279-283. doi:10.3760/cma.j.issn.2095-2848.2013.04.011

Objective To study the feasibility of a novel probe 99Tcm-HYNIC-2(poly-(ethylene glycol),PEG) 4-Dimer (Dimer:E-[c (RGDfK) 2]) as a potential imaging agent for integrin αv β3 positive tumors,and also to observe the influence of an angiogenesis inhibitor,endostar,on the biodistribution and tumor uptake of the tracer in tumor bearing nude mice.Methods The expression of integrin αv β3 in human glioma cells U87MG was determined with immunofluorescence staining before and after treatment with endostar.99Tcm-HYNIC-2PEG4-Dimer was prepared and administered in U87MG tumor bearing mice in 6 h after either administration of endostar (200 μl) or saline (control group) and then biodistribution study was performed.Other 16 mice were divided into endostar treated group (20 mg/kg) and control group (saline) and then gamma imaging was performed in the two groups.Statistical significance of differences between the two groups was assessed using two-sample t test.Results Radiochemical purity of 99Tcm-HYNIC-2PEG4-Dimer was exceeded 95％.The expression of integrin αvβ3 in U87MG cell was high and gradually decreased after treatment with endostar.There was a negative dose-effect relationship between the dose of endostar and the expression of integrin αvβ3 with the peak effect at the dose of 400 μg/ml.The distribution study in vivo showed that the tracer uptake of U87MG tumors was high,but it decreased after injection of endostar.At 90 min,the ％ID/g of endostar and control groups were 1.50±0.08 and 6.27±0.33,respectively (t =40.23,P＜0.05).The average T/NT ratios of 99Tcm-HYNIC-2PEG4-Dimer uptake in the endostar and control groups were 1.02±0.11 and 2.58±0.36,respectively (t =10.25,P＜0.05).The integrin αv β3 positive expression ratios of tumor in endostar and control groups were (33.1 ±2.7) ％ and (81.5±3.2) ％,respectively (t =32.60,P＜0.05).Conclusions The novel probe 99Tcm-HYNIC-2PEG4-Dimer may be a promising radiotracer for integrin αvβ3-positive tumor imaging.It may be used for monitoring the therapeutic effect of endostar and may be potentially used for screening the candidates of anti-angiogenesis therapy.

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The preparation of 99Tcm-J591 and its SPECT imaging of nude mice bearing human prostate cancer

Shaohua TU ; Jiangfan SHEN ; Rong TAO ; Xiaowen JI ; Yancheng WANG

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(4):284-288. doi:10.3760/cma.j.issn.2095-2848.2013.04.012

Objective To study the binding performance of 99Tcm labeled anti-human prostatic specific membrane antigen (PSMA) monoclonal antibody J591 (99Tcm-J591) and prostate cancer cells in vitro,the biodistribution and SPECT imaging of 99Tcm-J591 in nude mice bearing human prostate cancer in vivo.Methods The monoclonal antibody J591 was labeled with 99Tcm by improved Schwarz method.Labeled antibody was purified by Sephadex G-50.The labeling efficiency and radiochemical purity were measured by paper chromatography and trichloroacetic acid method.The binding performance of J591 and prostate cancer cells was measured by flow cytometry in vitro.The nude mice bearing PSMA-positive C4-2 prostate carcinoma xenografts served as experiment group,mice bearing PSMA-negative PC3 tumors served as control group.6.2-8.5 MBq of 99Tcm-J591 (25 μg) was intravenously injected into mice.Gamma imaging was performed 2,6,12 and 24 h after injection,T/NT was calculated by ROI technique.After scanned 12 h post injection,4 mice of the experiment group and 5 mice of the control group were sacrificed and the tracer in vivo biodistribution was measured by gamma-counting,and the ％ ID/g was calculated.Two-sample t test was carried out to validate significant difference of ％ID/g between two groups.Results The labeling efficiency and radiochemieal purity of 99Tcm-J591 were (78.9±6.2)％ and (92.3±5.1)％,respectively,and the specific activity of 99Tcm-J591 was 68.7 MBq/mg.The antibody J591 and 99Tcm-J591 could strongly combine with PSMA-positive C4-2 cells in vitro,and didn't combine with PSMA-negative PC3 cells in vitro.SPECT imaging results showed that radioactive concentration was obvious in tumor 6 h post injection,the concentration scope became large and the tumor image was clear 12 h post injection.T/NT was 1.9±1.1 at 2 h,4.3±1.8 at 6 h,5.6±2.7 at 12 h,1.4±0.6 at 24 h,respectively.In the control group,no radioactivity concentration was found in tumor,and T/NT was less than 2.The biodistribution results showed that ％ID/g of tumor tissue was 20.1±5.2 in the experiment group and 5.8±2.6 in the control group,and there was significant difference (t=5.37,P＜0.001).No significant tracer uptake occurred in other tissues and organs between the two groups (all t＜ 1.98,all P＞0.05).Conclusion The immunoactivity,characteristics of biodistribution and tumor targeting property of monoclonal antibody J591 show promising future and potential values in diagnosis and therapy of prostate cancer.

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Manufacture and application of a microfluidic chip-based 18F microreactor

Yongping ZHANG ; Yingjian ZHANG ; Mingwei WANG ; Center PET

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(4):298-302. doi:10.3760/cma.j.issn.2095-2848.2013.04.015

Objective To develop a polydimethylsiloxane (PDMS) microfluidic chip-based 18F microreactor for the preparation of 18F-labeled probes.Methods The 18F microreactor was composed of PDMS microfluidic chip and customized glass microvessel integrating with stainless capillary tube (D 0.6 mm) as heater/cooler.PDMS chips were fabricated by silk-screen printing technology,a traditional and easily accessible process.18F-FDG and 18F-fluoroacetate(FAC) were synthesized using the 18F microreactor.TLC was applied to measure the 18F-labeling yield and the radiochemical purity of 18F-FDG and 18F-FAC.Results The size of PDMS chip was 40.0 mm (l)×30.0 mm (w)×6.0 mm(h) and the liquid/gas inside channel was 0.3 mm (w)×50.0 μm (h).The customized glass microvessel was about 4.0 mm (D) ×30.0 mm (h) with 200 μl of reaction volume.The capillary tube which wrapped around the microvessel functioned as a heater when electric current was provided,while as a cooler when compressed air went through.The integrated 18F microreactor with a total size of 40.0 mm (l) ×30.0 mm (w) ×15.0 mm (h) was successfully used to prepare 18F-FDG and 18F-FAC,whose radiochemical purity were both higher than 96％ and 18F-labeling yield was 92.5％ and 90.0％ respectively in the first fluorination step.Conclusions A PDMS microfluidic chip-based 18F microreactor is developed and successfully applied to prepare 18F-FDG and 18F-FAC.It has the dual advantages of both microfluidc chip and traditional synthesis module and features of high integration,small total size and low consumption of labeling precursor.

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The clinical application of 18F-FDG PET/CT scan in the thyroid carcinoma

Liyan LU ; Hankui LU

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(4):312-315. doi:10.3760/cma.j.issn.2095-2848.2013.04.022

The incidence of thyroid cancer is the top ranking among endocrine carcinoma worldwide.Many imaging modalities have been applied in diagnosing,characterization of the biological behaviors and predicting the outcomes of various thyroid carcinoma.Over the years,18F-FDG PET/CT has been largely used to identify undifferentiated thyroid carcinoma cells in thyroid carcinoma patients with or without 131I avid lesion.The purpose of this mini-review was to update the clinical role and positive impact of 18F-FDG PET/CT in various thyroid carcinoma patients.

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Study on the diagnosis and mechanism of hemispatial neglect using different imaging modalities

Lele HUANG ; Yafu YIN ; Yaming LI

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(4):315-318. doi:10.3760/cma.j.issn.2095-2848.2013.04.023

Hemispatial neglect is a kind of cognitive impairment characterized by failure to report,respond or orient to stimuli presented in ipsi-or contra-lateral space after brain injury,which cannot be attributed to sensory or motor defects.Although there are various diagnosis and assessment methods,none is regarded as standard.The paper and pencil test is commonly used in routine clinical workup.The mechanism is controversial and the most accepted opinion is due to visuospatial attention defect.Imaging study focuses on exploring the functional regions leading to the hemispatial neglect.As functional imaging,SPECT and PET have capability to evaluate the perfusion,glucose metabolism and cellular vitality,which have been found increasingly useful and applicable clinically in the field of diagnosis for cognitive impairment diseases.They are likely to have potential for contribution in the study of hemispatial neglect.

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Clinical value of 18F-FDG PET/CT in the diagnosis of primary peritoneal papillary serous carcinoma

Zhichun LIN ; Liang YIN ; Tao HE ; Dongju ZHANG ; Qing ZHANG ; Haiyu MU

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(5):324-327. doi:10.3760/cma.j.issn.2095-2848.2013.05.002

Objective To explore the diagnostic value of 18F-FDG PET/CT in primary peritoneal papillary serous carcinoma (PPPSC).Methods Ten postmenopausal female cases of pathologically diagnosed PPPSC from March 2009 to October 2011 were retrospectively reviewed (age range:61-81 years,mean:(69.4±6.2) years).All cases underwent 18F-FDG PET/CT.The CT characteristics and SUVmax of lesions on PET images were analyzed.Serum CA125 levels were measured before or after PET/CT within one week.The patterns of PPPSC on PET/CT were compared with histopathological results.Linear correlation analysis was used to evaluate the correlation between the CA125 and the maximum SUVmax of lesions presented in parietal peritoneum,greater omentum or mesentery.Results The PET/CT uptake pattern of the 10 PPPSC cases was described as floccus,multi-nodular or cake-like in greater omentum (SUVmax =6.32±2.87),and as diffuse or localized nodules,or non-uniform strip-like thickening in 9 parietal peritoneum and 8 mesentery cases (SUVmax =5.96±2.14 and 5.70± 1.69,respectively).The most commonly involved sites were pelvic wall of peritoneum and mesentery of small intestine.All 10 cases had different degrees of ascites,mainly intrapelvic and perihepatic.Hypermetabolic ovarian enlargement (all ＜5 cm) was bilateral in 2 patients and right-sided in 1 patient.Four patients had retroperitoneal lymph node metastasis and others were found with punctate calcifications in metastatic lymph nodes,small pleural effusions,liver metastasis,as well as portal node metastasis.CA125 concentration was elevated in all cases ((51.25±26.40) ×104 U/L),but there was no significant correlation between CA125 and the maximum SUVmax of lesions found in parietal peritoneum,greater omentum or mesentery (r=0.05,P＞0.05).Conclusion 18F-FDG PET/CT could show the positions and metabolic status of PPPSC lesions.It may be an effective imaging modality in the diagnosis and assessment of PPPSC.

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The value of 18F-FDG PET/CT in transcatheter hepatic arterial chemoembolization therapy of primary hepatocellular carcinoma with high FDG uptake

Yanli WANG ; Na FANG ; Lei ZENG ; Xinjian CUI

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(5):328-331. doi:10.3760/cma.j.issn.2095-2848.2013.05.003

Objective To evaluate the therapeutic response of transcatheter hepatic arterial chemoembolization (TACE) for primary HCC using 18 F-FDG PET/CT.Methods Twenty-four HCC patients (21males,3 females; age range:40 to 75 years,mean age:58.1 years) with high uptake of 18F-FDG on pretreatment PET/CT and with follow-up 18F-FDG PET/CT 1-2 months post TACE were retrospectively analyzed.A lesion was regarded as positive for HCC if it had higher FDG uptake than the surrounding liver parenchyma.All patients were followed for 3 months by clinical symptoms,AFP and 18F-FDG PET/CT.x2 test was used to analyze the difference between the diagnostic value of 18F-FDG PET/CT and AFP.Results On follow-up 18F-FDG PET/CT,9 patients had lower 18F-FDG uptake than the surrounding liver parenchyma,indicating no residual tumor; while 11 patients with residual tumor showed higher uptake of FDG than the liver background though the uptake was lower than that of baseline PET/CT.Two patients with isometabolic FDG uptake were confirmed of having residual tumor during follow-up.One case showed increased 18F-FDG uptake more than that of baseline,indicating disease progression.One case was false positive for residual tumor due to over attenuation correction in the presence of lipiodol.The sensitivity and accuracy for the detection of residual tumor by 18F-FDG PET/CT and AFP were 100％ (14/14) vs 71.4％ (10/14),and 95.8％(23/24) vs 70.8％(17/24) (x2 =6.56 and 4.18,both P＜0.05),respectively.Conclusion 18F-FDG PET/CT is useful to evaluate the efficacy of TACE for 18F-FDG-avid HCC and it may provide important information for further treatment planning.

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Clinical value of 18F-FDG PET/CT in the management of Castleman's disease

Zhanli FU ; Xuchu ZHANG ; Yan FAN ; Jianhua ZHANG ; Rongfu WANG

Chinese Journal of Nuclear Medicine and Molecular Imaging.2013;33(5):332-335. doi:10.3760/cma.j.issn.2095-2848.2013.05.004

Objective To assess the value of 18F-FDG PET/CT in clinical classification,monitoring of chemotherapeutic response and surveillance of histopathological transformation of Castleman's disease (CD).Methods Fourteen pathologically diagnosed CD patients (7 males,7 females; mean age:(45.64±14.30) years) were retrospectively reviewed.18F-FDG PET/CT was performed before chemotherapy in all patients and 4 of 14 patients were reexamined after the treatment.The study parameters included histopathological results,sites,number and highest SUVmax of the lesions.Mann-Whitney and Kruskal-Wallis tests were used for data analysis.Results Of all the 12 patients without histopathological transformation,one or more enlarged and metabolically active lymph nodes were found in each patient (SUVmax =3.94± 1.44,range:1.9-6.8),including 2 unicentric CD (UCD) and 10 multicentric CD (MCD).There was no statistically significant difference of SUVmam between UCD and MCD (4.55±3.18 vs 3.82±1.14; Z=0.22,P＞0.05).There was also no significant difference of SUVmax among different pathological types (hyaline-vascular CD (4/12):3.56±0.96,plasma cell CD (6/12):4.73±1.41,mixed CD (2/12):2.30±0.57; x2 =4.74,P＞0.05).For the 4 patients with follow-up PET/CT after chemotherapy,the lesion activity was normalized in 3 patients and clearly reduced in 1 patient.The SUVmax of 2 patients with histopathological transformation (10.85±2.05) was significantly higher than that without transformation (3.94± 1.44; Z=-2.19,P＜0.05).Conclusion 18F-FDG PET/CT may play an important role in clinical classification,monitoring of chemotherapeutic response and surveillance of histopathological transformation of CD.

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