Objective To construct the human IL-24 recombined adenovirus(Ad-IL24)and to observe the effect of Ad-IL24 on ex vivo culture of HL-60 cells.Methods pAdTrack-CMV-IL24 was constructed by PCR with pcDNA3.0-IL24 recombined plasmid as a template,enzyme digestion and ligation.The pAdTrack-CMV-IL24 lineared by Pme Ⅰ was co-transformed into BJ5183 with pAdEasy-1.The pAdEasy-1-pTrack-CMV-IL24 recombined adenovirus vector was lineared with Pac Ⅰ and then transfected in to QBI-293A cells.The Ad-IL24 was obtained and used to infect HL-60 ceils,and the effect on HL-60 cells was tested by LSCM,Mrrr,FCM,ELISA and immunohistochemistry staining.Results The pAdEasy-1-pTrack-CMV-IL24 vector was constructed and the Ad-IL24 was successfully obtained.The effect of inhibiting growth of HL-60 cells and inducing cell apoptosis by IL-24 was proved by LSCM,MTT and FCM.IL-24 down-regulated the expression of anti-apoptosis factor Bcl-2 and increased the secretion of IFN-γ,TNF-α of HL-60 cells.Conclusion Ad-IL24 can inhibit the growth of HL-60 cells and induce cell apoptosis through down-regulating expression of anti-apoptosis factor and increasing the secretion of IFN-γ,TNF-α of HL-60 cells.The human Ad-IL24 may provide a basic study for the future target therapy to tumors.

Objective:To construct the cells which can express LIF protein steadily and study the effect of rhLIF and IL-24 on HL-60 cells.Methods:ECV-304 cells were infected by eukaryotic expression plasmid pcDNA3-LIF,then the positive cells were selected by G418.The positive cells were collected and their culture supernatant was stored for further use.Expression of LIF mRNA was detected by RT-PCR.The pAdeasy-1-pAdTrack-CMV-IL-24 was extracted from DH5a.The recombined adenovirus vector was lineared with PacI and transfected into 293 cells.The IL-24 recombined adenovirus(Ad-IL-24) was obtained and used to infect HL-60 cells.At the same time,the culture supernatant containing rhLIF was added into the HL-60 cells which was identified as positive expression of IL-24 by RT-PCR.Synergistic effect of the cytokines on HL-60 cells was tested by LSCM, FCM and immunohistochemistry stain assay.Results:The cells expressing LIF protein steadily were constructed successfully, the high titer of the recombined adenovirus(Ad-IL-24) was obtained. Expression of IL-24 in infected HL-60 cells was identified by RT-PCR.The apoptosis of HL-60 cells induced by rhLIF and IL-24 was proved by LSCM ,FCM and immunohistochemistry stain assay. They had synergistic effect.Conclusion:rhLIF and IL-24 can inhibit growth of HL-60 cells and induce apoptosis of the cells.They have synergistic effect.

Objective:To compare the anti-androgenic effect of cyproterone acetate (CPA) and spironolactone (SPL) on male-to-female transsexuals.Methods:From January 2012 to September 2021, 185 male-to-female transsexuals (95 using CPA and 90 using SPL) who visited the Peking University Third Hospital and under stable medication for ≥3 months were enrolled, aged 16 to 40 (23±5) years. General information and laboratory indicators of the last visit were collected for a cross-sectional study.Results:The median doses of antiandrogens in the CPA group and SPL group were 25 mg/d and 80 mg/d, respectively. And the median dose of oral estradiol valerate in both groups was 2 mg/d. Testosterone level in the CPA group was significantly lower than the SPL group [0.7 (0.7-1.5) nmol/L vs. 13.2(6.7-18.4) nmol/L, U= 6 970.500, P<0.001]. The CPA group also had better subjective effects on testicular atrophy, erection decrease, body hair decrease, skin softening and figure feminization (all P<0.05). The prolactin level of CPA group was significantly higher than that of SPL group [21.5 (12.6-30.1) ng/ml vs. 11.9 (7.7-20.0) ng/ml, U= 2 053.500, P<0.001]. Conclusions:CPA has a more significant effect on lowering testosterone levels than SPL, and is better than SPL in terms of testicular atrophy, erection decrease, body hair decrease, skin softening and figure feminization, albeit with a potentially higher risk of hyperprolactinemia.