ABSTRACT
Objective To investigate the changes of iodide uptake and the expression of thyroidspecific genes in poorly differentiated follicular thyroid carcinoma (FTC) cells after transfection of human TSH receptor (hTSHR) gene in vitro. Methods The recombinant eukaryotic expression plasmid PcDNA3. 1/hTSHR-cDNA was transformed into DH5a bacterial for amplification and then the recombinant plasmid was extracted. The recombinant was identified with PCR amplifying, restriction enzyme digestion analysis and DNA sequencing. The recombinant plasmid pcDNA3.1/hTSHR was transfected into FTC-133 cell line by lipofectin methodin vitro. Immunofluorescence, iodide uptake studies and real time-PCR were applied to detect target protein expression. Statistical analysis was performed with t-test using SPSS 13. 0 software. Results Kpn Ⅰ and Xba Ⅰ restriction enzyme digestion, PCR amplifying and DNA sequencing confirmed that pcDNA3. 1/hTSHR was successfully constructed. After transfection of the recombinant plasmid pcDNA3. 1/hTSHR-cDNA and the stimulation of hTSH, the tumor cells displayed the expression of hTSHR protein at cell surface and cytoplasm. The iodine uptake in pcDNA3. 1/hTSHR transfected cells was 2. 9 times higher than that of control(pcDNA3.1(+) transfected cells) group(t = 28.63, P ＜0. 01). The expression of TSHR,NIS, TPO and Tg (mRNA levels) in pcDNA3. 1/hTSHR transfected cells were also significantly elevated by 1.74 (t =5.959, P＜0.01), 7.2 (t =3.807,P＜0.05), 2.88 (t=4.769,P＜0. 01) and 2.67 times (t=6.388,P ＜0.01) respectively compared to those of the control group. Conclusion The study demonstrates that iodide uptake may be reactivated by hTSHR receptor gene transfection in poorly differentiated FTC cell.
ABSTRACT
Objective To investigate the clinical value of ~(99)Tc~m-ethylenedicysteine (EC) diuretic renography (DR) in pre-and post-operative pediatric congenital hydronephrosis.Methods The DR with injection of Furosemide at 15 min of forty children with hydronephrosis was retrospectively studied.The preoperative renal blood perfusion rate (BPR),effective renal plasma flow (ERPF),grade of hydronephrosis,renogram and renal dynamic imaging of pre- and post-operative kidneys were compared.The t-test and Mann-Whitney test were used for data analysis.Results (1) Of 40 pathological kidneys,the BPR increased 5.99% (t=-5.13,P＜0.01)from pre-operative to post-operative:(34.05±11.07)% to (40.04±8.56)%.The ERPF increased 12.48 ml/min(t=-4.35,P＜0.01) from pre-operative to post-operative:(57.81±34.32)ml/min to(70.29±5.37)ml/min.(2)The grade of hydronephrosis of 40 pathological kidneys improved significantly(Z=-2.64,P＜0.01) with the mean sum of ranks of 47.21 pre-operatively to 33.79 post-operatively.(3) As the hydronephrosis worsened,the collecting system became bigger,the renal parenchyma became thinner,the extent of intrarenal parenchymal photopenia became larger and the response to diuretic challenge in pathological kidneys decreased or became totally irresponsive.(4)Thirty-seven cases of obstruction at ureteropelvic junction (UPJO) and 3 cases at ureterovesical junction (UVJO) were diagnosed by DR,which were all confirmed by surgery.Conclusions DR is a reliable method to evaluate pediatric congenital hydronephrosis.It can accurately reflect the grade and (or) severity of the disease,guide therapy and assess the therapeutic success of operation.