The aim of this study was to establish stable expression of human thyroid stimulating hormone receptor (TSHR) alpha-subunit (hTSHRA) on human embryonic kidney 293T (HEK 293T). HEK 293T cell lines with stable expression of hTSHRA could be used for detecting affinity between hTSHRA and potential TSHR blocking-peptide. We firstly constructed hTSHRA gene into lentiviral vectors GV218. The sequence comparison indicated that we had constructed GV218-hTSHRAA. Western blot demonstrated the 52 kD aim band of hTHSRA on over-expressed HEK 293T cells. GV218-hTSHRA constructions and pHelper were then co-transfected into HEK 293T cells to form packaging plasmid. The HEK 293T cells that stably expressed hTSHRA could also express green fluorescent pro tein. The titer of lentiviral packaging vector is 2 x 10(8) TU/mL with qPCR. The lentiviral packaging vector thereafter was transfected into HEK 293T cells again. The hTSHRA expressed on the HEK 293T cells. Human TSHRA stab ly expressed on HEK 293T upon continuously passaging. Therefore, we established hTSHRA stable expression on HEK 293T cells by constructing GV218-hTHSR lentiviral packaging vector. It is a useful tool for studying TSHR affinity with anti-thyroid peptide.
Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; Plasmids ; Receptors, Thyrotropin ; biosynthesis ; Transfection

Objective To compare the diagnostic values of 18F-fluorodeoxyglucose-single photon emission computed tomography (18F-FDG-SPECT) and helical CT in the detection of the metastasis of postoperative breast cancer.Methods A total of 94 patients with postoperative breast cancer were chosen as research objects.The follow-up duration of post-operation was 2 years.All patients were received by 18F-FDG-SPECT and helical CT during follow-up.Lymph nodes that were suspected to the postoperative metastasis were taken for histological procedure.The diagnostic values of 18F-FDG-SPECT and helical CT to the metastasis of postoperative breast cancer were compared.Results Compared to helical CT,the sensitivity,specificity,positive predictive value,negative predictive value,and positive likelihood ratio of 18F-FDG-SPECT were higher (P ＜ 0.05);however,the negative likelihood ratio of 1s F-FDG-SPECT were lower (P ＜ 0.05).Conclusions 18F-FDG-SPECT has more important clinical value in the detection of metastasis of postoperative breast cancer relative to helical CT.

Colorectal cancer is a leading cancer in many countries. The glucose metabolic activities of the tumor cells are higher than that of the normal cells. They could uptake much more 2-[18F] Fluoro-2-Deoxy-d-Glucose (18F-FDG), an analogue of glucose, than normal cells do. The physicians can search the lesions via 18F-FDG positron emission tomography (PET). 18F-FDG-PET has higher sensitivity and accuracy in diagnosing the primary lesion, recurrence, liver metastasis and extrahepatic metastasis of the colorectal cancer, compared with the conventional technique. The results of the 18F-FDG-PET often restage the colorectal cancer and even change the treatment plan as 18F-FDG-PET provides more information than traditional technique does. Also, it could be used to follow up the case of the colorectal cancer after operation, systematic chemotherapy and radiotherapy. The current review addresses the metabolism, the clinical application, the shortcomings and the future prospect.
Colorectal Neoplasms ; diagnostic imaging ; Fluorodeoxyglucose F18 ; Humans ; Positron-Emission Tomography ; Radiopharmaceuticals

Objective To explore clinic value of nuclear myocardiac perfusion imaging (MPI) in perioperative coronary risk assessment in patients with heart diseases planning arthroplasty,and compare the conformity and effect on surgery planning between MPI and coronary computed tomography angiogram (CTA).Methods Prospectively collect clinical data of 228 patients with heart diseases planning joint arthroplasty,all patients had MPI and CTA test to assess perioperative coronary risk.The conformity and effect on surgery planning between the MPI and CTA was analyzed,and the occurrence of cardio-vascular complication was recorded.Results MPI showed that 215 patients had no myocardial ischemia except for 13.CTA showed that 113 cases had no coronary stenosis,51 cases had light coronary stenosis,39 cases had middle coronary stenosis and 12 cases had severe coronary stenosis in patients with no myocardial ischemia.CTA also found that 3 cases had no coronary stenosis,4 cases had light coronary stenosis,3 cases had middle coronary stenosis and 3 cases had severe coronary stenosis in patients with myocardial ischemia.The conformity between MPI and CTA was low.222 patients had completed operation except for 6 cases.MPI rather than CTA had conspicuous effect on surgery planning.There was no angina,myocardial infarction or cardiac death associated with myocardial ischemia.The postoperative cardio-vascular complications included 6 cases of heart failure,4 cases of atrial fibrillation and 1 case of cerebral infarction.Conclusion MPI is a direct index to judge blood supply of the myocardium,which can exactly predict the risk of perioperative myocardial ischemia and the prognosis.It's helpful for preoperative planning,reducing risk of perioperative complications through screening and excluding high-risk patients.

Human Sodium/Iodide symporter gene cDNA was amplified from thyroid tissue of the patient suffering from Graves disease by RT-PCR, and T/A cloned into pGEM-TEasy-NIS for sequencing, subcloned into shuttle plasmid pAdTrack-CMV which contained a green fluorescent protein (GFP) gene, and then forwarded to homologous recombinant in the bacteria BJ5183 that already contained AdEasy-1 plasmid. Positive recombinant adenovirus vector was selected, packaged and amplified in the 293 cells to obtain recombinant adenovirus. The results showed that the recombinant AdNIS was correctly constructed and confirmed by restriction enzyme analysis and PCR. The viral titer was 2. 5 - 3 x 10(9) efu/ml. So, the recombinant adenovirus vector carrying hNIS was successfully constructed, thus providing a basis for researches on 131I therapy in nonthyroid carcinoma.
Adenoviridae ; genetics ; metabolism ; DNA, Complementary ; genetics ; Genetic Vectors ; genetics ; metabolism ; Graves Disease ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Symporters ; biosynthesis ; genetics

The aim of this study was to assess the role of 131 I-L-tyrosine for diagnosing E. coli infection and differentiating the infection from the inflammary lesion in mice. The L-tyrosine was labeled with 125 I by N-bromosuccinimide. The binding rates of 125 I-L-tyrosine with E. coli were tested by culturing E. coli with 125 I-L-tyrosine in poor nitrogen yeast medium and in rich nitrogen LB medium separately. The bio-distribution of 125 I-L-tyrosine and the 131 I-L-tyrosine SPECT scintigraphy in E. coli infection of mice were studied separately. The results revealed that the label rate of 125 I-L-tyrosine was more than 99%. The percentage of 125 I-L-tyrosine binding with E. coli was 2.78% in yeast medium, and was 0.85% in LB medium. The distribution peak of 125 I-L-tyrosine in E. coli focal infection of mice was in the range between 45 and 60 min. The infection/normal muscle ratio was 2.46 after 60 minutes. The SPECT scintigraphy demonstrated an accumulation of the 131 I-L-tyrosine in the E. coli focal infection during the 45-60 minutes. The infection/normal muscle ratio of the 131 I-L-tyrosine radioactivity was 2.51 in infectious mouse. However, the SPECT scintigraphy also demonstrated an accumulation of the 131 I-L-tyrosine in the inflammatory lesion during the 45-60 minutes. The lesion/normal muscle ratio of the 131 I-L-tyrosine radioactivity was 2.29 in inflammatory mouse at 60 mins. Thus, the 131 I-L-tyrosine SPECT scintigraphy could diagnose the E. coli infection, but it can not be used for differentiating the bacteria infection and the inflammatory lesion in mice. Our study revealed that the 131 I-L-tyrosine would play a potential role in diagnosing the inflammatory lesion in human beings.
Animals ; Diagnosis, Differential ; Escherichia coli Infections ; diagnostic imaging ; Inflammation ; diagnostic imaging ; Iodine Radioisotopes ; Mice ; Tomography, Emission-Computed, Single-Photon ; methods ; Tyrosine

This paper was aimed to investigate the biodistribution and ability of free 131-bcl-2/bcl-xl ASON (FA) and anionic long circulation liposomes encapsulated with 131I-bcl-2/bcl-xlASON (NA), in tumor-bearing rats, to image breast cancer. We investigated the tissue distribution of NA in virgin female Sprague-Dawley (SD) rats with n-methyl nitrosourea (MNU)-induced breast cancers in situ. The percentage of the injected dose per gram (%ID/g) was calculated, with the maximum ratios of tumor to blood and tumor to muscle, after injections of NA and FA for 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, 12 h and 24 h, respectively. The ability of NA to image breast cancer in tumor-bearing rats was determined using emission computed tomography (ECT). Seventy percent (90/130) SD rats in the study developed mammary tumors after MNU injection with the average latency (NA) (96 +/- 1.2)days. The %ID/g of NA in breast cancer tissue, tumor bearing rats in liver and spleen tumor tissues after 10 hours were (6.23 +/- 0.23) %ID/g, (12.00 +/- 0.26) %ID/g and (18.25 +/- 1.33)% ID/g, respectively. The ratios of tumor to blood 6.29 +/- 0.76 and tumor to muscle 10.55 +/- 0.68 in tumor bearing rats slowly maximized at 10 h post injection of NA, most probably due to the enhanced permeability and retention effect. Hence in radionuclide antisense scintigraphy, the breast cancer in rat was clearly displayed at 10h after iv administration of NA-D. However, tumors were not visualized in rats with the iv injection of NS and NN even at the delayed time. Due to the inhibition of rapid uptake of NA by the reticulo-endothelial system, NA displays valuable pharmacologic properties characterized by the enhanced accumulation in tumor.
Animals ; Female ; Iodine Radioisotopes ; Liposomes ; pharmacokinetics ; Mammary Neoplasms, Experimental ; metabolism ; Oligonucleotides, Antisense ; pharmacokinetics ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Radionuclide Imaging ; methods ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution ; bcl-X Protein ; metabolism

The first aim of this study was to compare the serum thyroglobulin (Tg) positive rate between differentiated thyroid carcinoma (DTC) patients with positive thyroglobulin antibody (TgAb) and patients with negative TgAb. The second aim of this study was to investigate the correlation between serum Tg value and antithyroglobulin (TgAb) concentration of patients with DTC. We collected the serum Tg value and TgAb concentration of patients with DTC after thyroid ablation by operation and radioiodine therapy retrospectively. Then we investigated the Tg positive rate of DTC patients with positive TgAb and patients with negative TgAb separately. The scatter diagram between serum Tg value and TgAb concentration of DTC patients was performed to analyze their potential relationship. As a result, among 252 patients with DTC after thyroid ablation, 7 of 47 patients (14.89%) with positive TgAb (>115 IU/mL) had positive serum Tg (>10 microg/L), and 61 of 205 (29.76%) patients with negative TgAb (<115 IU/mL) had positive serum Tg. Eighty three of the 252 patients with DTC had accurate serum Tg and TgAb concentration. No correlation between serum Tg value and TgAb concentration was found among these Eighty three patients. Thus, it may be concluded that positive TgAb could cause much more TgAb interference in serum Tg value than negative TgAb could because of a different quality of TgAb. Serum Tg value is not correlated with serum TgAb concentration in patients with DTC after thyroid ablation.
Adolescent ; Adult ; Aged ; Autoantibodies ; blood ; Carcinoma ; blood ; immunology ; pathology ; Female ; Humans ; Male ; Middle Aged ; Thyroglobulin ; blood ; Thyroid Neoplasms ; blood ; immunology ; pathology ; Young Adult

Humans ; Pilot Projects ; Prospective Studies ; Pulmonary Emphysema/surgery* ; Lung/surgery* ; Emphysema ; Perfusion ; Tomography, Emission-Computed ; Tomography ; Tomography, Emission-Computed, Single-Photon ; Pneumonectomy ; Bronchoscopy/methods*