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Aim To express and purify recombinant hCGH-CTP fusion protein in high-density suspension culture of Chinese hamster ovary cells (CHO-S), and to verify the lipid accumulation effect of rhCGH-CTP on 3T3-L1 mature adipocytes. Methods The recombinant protein expression vector (pcDNA3. 1-rhCGH-CTP) was constructed, achieved by fusing the human glycoprotein hormone beta 5/alpha 2 cDNA with CTP Linker. The expression plasmid was transiently transfected into the suspended CHO-S to express rhCGH-CTP protein and then purified, and the protein biological activity was verified. Intervention with 3T3-L1 mature adipocyte cells for 24 h was performed to detect the changes of intracellular triglyceride (TG) level. Results Western blot results showed that rhCGH-CTP protein was successfully expressed in CHO-S cells, and the yield was up to 715. 4 mg • L~ . The secreted protein was purified by AKTA pure system with higher purity that was up to 90% as identified by SDS-PAGE. In addition, the intracellular cAMP content of mature adipocytes with high expression of TSHR gene significantly increased after intervention with different concentrations of rhCGH-CTP protein by ELISA kit, indicating that rhCGH-CTP protein had biological activity. Oil red 0 staining showed that compared with the control group, the lipid content of mature adipocytes in the intervention groups with different concentrations of rhCGH-CTP protein significantly decreased (P < 0. 05) . Conclusions The rhCGH-CTP protein has been successfully expressed and purified with biological activity, and effectively reduce TG. This research provides an important theoretical basis for further revealing the physiological role of CGH protein and its potential application in clinical practice.
RESUMO
Objective To evaluate the performance of energy discriminating dosimeter and compare the impact of the assessment results. Methods CTLD-J4000 energy discriminating dosimeter was irradiated with different energy photons, and the differences covered by the dosimeters were used. Density filtering materials have different filtering effects on photons, and the ratio curve of the readings to the photon energy was fitted to identify the photon energy. Results The coincidence between the non-filter/light filter of the CTLD-J4000 energy discrimination dosimeter and the manufacturer's reference value was better than that of the non-filter/re-filter. The latter had higher deviation values at 48 keV(12.8%)and 83 keV(15.5%), respectively. Using the average scale factor, the maximum energy response deviation was 23.5%. Conclusion Using a discriminating dosimeter can better identify the ray energy, thereby reducing the energy response error caused by using the average scale factor.