Objective： To construct the recombinant adenovirus vector carrying rat vascular endothelial growth factor（VEGF）, as preparation for genetic transfection that follows. Methods： Rat VEGF was obtained by using RT-PCR...Objective： To construct the recombinant adenovirus vector carrying rat vascular endothelial growth factor（VEGF）, as preparation for genetic transfection that follows. Methods： Rat VEGF was obtained by using RT-PCR amplification and then cloned into the shutter plasmid pDC316. Subsequently, this newly constructed plasmid pDC316-VEGF, after identification by nuclease digestion analysis and sequencing analysis, was transfected into human embryonic kidney cells HEK293 by Lipofectamine 2000 mediation, together with adenovirus-packaging plasmid pBHGE3. Based on the homologous recombination of the two plasmids within HEK293 cells, the recombinant adenovirus vector carrying VEGF and VDC316-VEGF was created. VDC316-VEGF was subsequently identified using PCR, purified using repeated plaque passages, proliferated using freezing and melting within HEK293 cells, and titrated using 50% Tissue Culture Infective Dose（TCID50） assay. Results：The newly constructed recombinant adenovirus was confirmed to carry rat VEGF based on PCR results, and its titration value determined based on TCID50 assay was 3 × 10^9 pfu/ml. Conclusion：The recombinant adenovirus carrying rat VEGF was successfully constructed. The newly constructed adenovirus can produce a ufficiently high titration value within HEK293 cells, providing a reliable tool for genetic transfection in further gene therapy researches.展开更多
Background： The first and most important step in characterizing familial nonmedullary thyroid carcinoma （NMTC） is to distinguish the true familial patients, which is the prerequisite for all accurate analyses. This...Background： The first and most important step in characterizing familial nonmedullary thyroid carcinoma （NMTC） is to distinguish the true familial patients, which is the prerequisite for all accurate analyses. This study aimed to investigate whether patients from families with ≥3 first-degree relatives affected with NMTC have different characteristics than patients from families with only two affected members, and to compare these patients with those with sporadic disease. Methods： We analyzed the clinicopathological features and prognosis of 209 familial and 1120 sporadic cases of NMTC. Familial patients were further divided into two subgroups： families with two affected members and families with ≥3 affected members. Results： The familial group had a significantly higher risk of bilateral growth, multifocality, extrathyroidal extension, and lateral lymph node metastasis than the sporadic group （P 〈 0.05）. These main features were also different between the group with ≥3 affected members and the sporadic group. The only difference between the two affected members＇ group and the sporadic group was incidence of multifocality （P 〈 0.05）. The probability of disease recurrence in patients from families with ≥3 affected members was significantly higher than that in sporadic cases （14.46% vs. 5.27%; P = 0.001）, while the probability in patients from families with two affected members was similar to that in sporadic patients （6.35% vs. 5.27%; P = 0.610）. The Kaplan–Meier survival analysis showed a statistically significant difference in disease-free survival between the two subgroups （85.54% vs. 93.65%; P = 0.045）. Conclusions： Patients from families with ≥3 members affected by NMTC have more aggressive features and a worse prognosis than those from families with only two affected members. Patients from families with ≥3 affected first-degree relatives may be considered to have true familial NMTC.展开更多
基金Natural Science Foundation of Jiangsu Province(BK2005158)
文摘Objective： To construct the recombinant adenovirus vector carrying rat vascular endothelial growth factor（VEGF）, as preparation for genetic transfection that follows. Methods： Rat VEGF was obtained by using RT-PCR amplification and then cloned into the shutter plasmid pDC316. Subsequently, this newly constructed plasmid pDC316-VEGF, after identification by nuclease digestion analysis and sequencing analysis, was transfected into human embryonic kidney cells HEK293 by Lipofectamine 2000 mediation, together with adenovirus-packaging plasmid pBHGE3. Based on the homologous recombination of the two plasmids within HEK293 cells, the recombinant adenovirus vector carrying VEGF and VDC316-VEGF was created. VDC316-VEGF was subsequently identified using PCR, purified using repeated plaque passages, proliferated using freezing and melting within HEK293 cells, and titrated using 50% Tissue Culture Infective Dose（TCID50） assay. Results：The newly constructed recombinant adenovirus was confirmed to carry rat VEGF based on PCR results, and its titration value determined based on TCID50 assay was 3 × 10^9 pfu/ml. Conclusion：The recombinant adenovirus carrying rat VEGF was successfully constructed. The newly constructed adenovirus can produce a ufficiently high titration value within HEK293 cells, providing a reliable tool for genetic transfection in further gene therapy researches.
文摘Background： The first and most important step in characterizing familial nonmedullary thyroid carcinoma （NMTC） is to distinguish the true familial patients, which is the prerequisite for all accurate analyses. This study aimed to investigate whether patients from families with ≥3 first-degree relatives affected with NMTC have different characteristics than patients from families with only two affected members, and to compare these patients with those with sporadic disease. Methods： We analyzed the clinicopathological features and prognosis of 209 familial and 1120 sporadic cases of NMTC. Familial patients were further divided into two subgroups： families with two affected members and families with ≥3 affected members. Results： The familial group had a significantly higher risk of bilateral growth, multifocality, extrathyroidal extension, and lateral lymph node metastasis than the sporadic group （P 〈 0.05）. These main features were also different between the group with ≥3 affected members and the sporadic group. The only difference between the two affected members＇ group and the sporadic group was incidence of multifocality （P 〈 0.05）. The probability of disease recurrence in patients from families with ≥3 affected members was significantly higher than that in sporadic cases （14.46% vs. 5.27%; P = 0.001）, while the probability in patients from families with two affected members was similar to that in sporadic patients （6.35% vs. 5.27%; P = 0.610）. The Kaplan–Meier survival analysis showed a statistically significant difference in disease-free survival between the two subgroups （85.54% vs. 93.65%; P = 0.045）. Conclusions： Patients from families with ≥3 members affected by NMTC have more aggressive features and a worse prognosis than those from families with only two affected members. Patients from families with ≥3 affected first-degree relatives may be considered to have true familial NMTC.
