Objective:To investigate the effect of Smac mimetic compound SM-164 on the radiosensitivity of cervical cancer cells and its underlying mechanisms.Methods:Human cervical cancer cell lines HeLa and SiHa were treated with different concentrations of SM-164 (0.05, 0.1, 0.2, 0.4 μmol/L) for 24 hours, followed by irradiation with 6 MV X rays at different doses (0, 2, 4, 6, 8 Gy). Clone formation assay was used to detect the survival fraction of HeLa and SiHa cells. The cells were divided into the negative control (NC) group, 4 Gy ionizing radiation (IR) group, 4 Gy combined with 0.2 μmol/L SM-164 treatment (IR+SM-164) group, and Caspases (Casp) broad-spectrum inhibitor Z-VAD blocking (IR+SM-164+Z-VAD) group. CCK-8 assay was adopted to assess cell proliferation, flow cytometry was used to detect cell apoptosis, and Western blot was employed to assess the changes in apoptosis-related proteins cleaved (C)-Casp9/Casp9, C-Casp3/Casp3, C-poly(ADP ribose) polymerase (PARP)/PARP, and DNA damage marker phosphorylated-histone H2A family member X (γH2AX)/H2AX ratios. One-way ANOVA was used for the comparison of means among multiple groups, and the SNK- q test was used for pairwise comparisons between two groups. Results:Compared with the NC group, the survival rates of HeLa and SiHa cells were significantly decreased after the combined treatment of IR and SM-164 (both P<0.01), and different concentrations of SM-164 enhanced cell radiosensitivity (all P<0.05). Compared with the NC group, cell proliferation activity was significantly decreased in the IR, IR+SM-164 and IR+SM-164+Z-VAD groups (all P<0.01), while apoptosis rate and the ratios of C-Casp9/Casp9, C-Casp3/Casp3, C-PARP/PARP, and γH2AX/H2AX were significantly increased (all P<0.01). Compared with the IR group, the proliferation activity in the IR+SM-164 group was significantly decreased ( P<0.05), whereas apoptosis rate and related protein ratios were significantly increased (all P<0.05). The γH2AX/H2AX ratio was increased in the IR+SM-164+Z-VAD group ( P<0.05). Compared with the IR+SM-164 group, the proliferation activity in the IR+SM-164+Z-VAD group was increased ( P<0.05), while the apoptosis rate and related protein ratios were significantly decreased (all P<0.05). Conclusion:SM-164 enhances the radiosensitivity of cervical cancer cells by inducing Casp activation.

Objective:To investigate the effect of Smac mimetic compound SM-164 on the radiosensitivity of cervical cancer cells and its underlying mechanisms.Methods:Human cervical cancer cell lines HeLa and SiHa were treated with different concentrations of SM-164 (0.05, 0.1, 0.2, 0.4 μmol/L) for 24 hours, followed by irradiation with 6 MV X rays at different doses (0, 2, 4, 6, 8 Gy). Clone formation assay was used to detect the survival fraction of HeLa and SiHa cells. The cells were divided into the negative control (NC) group, 4 Gy ionizing radiation (IR) group, 4 Gy combined with 0.2 μmol/L SM-164 treatment (IR+SM-164) group, and Caspases (Casp) broad-spectrum inhibitor Z-VAD blocking (IR+SM-164+Z-VAD) group. CCK-8 assay was adopted to assess cell proliferation, flow cytometry was used to detect cell apoptosis, and Western blot was employed to assess the changes in apoptosis-related proteins cleaved (C)-Casp9/Casp9, C-Casp3/Casp3, C-poly(ADP ribose) polymerase (PARP)/PARP, and DNA damage marker phosphorylated-histone H2A family member X (γH2AX)/H2AX ratios. One-way ANOVA was used for the comparison of means among multiple groups, and the SNK- q test was used for pairwise comparisons between two groups. Results:Compared with the NC group, the survival rates of HeLa and SiHa cells were significantly decreased after the combined treatment of IR and SM-164 (both P<0.01), and different concentrations of SM-164 enhanced cell radiosensitivity (all P<0.05). Compared with the NC group, cell proliferation activity was significantly decreased in the IR, IR+SM-164 and IR+SM-164+Z-VAD groups (all P<0.01), while apoptosis rate and the ratios of C-Casp9/Casp9, C-Casp3/Casp3, C-PARP/PARP, and γH2AX/H2AX were significantly increased (all P<0.01). Compared with the IR group, the proliferation activity in the IR+SM-164 group was significantly decreased ( P<0.05), whereas apoptosis rate and related protein ratios were significantly increased (all P<0.05). The γH2AX/H2AX ratio was increased in the IR+SM-164+Z-VAD group ( P<0.05). Compared with the IR+SM-164 group, the proliferation activity in the IR+SM-164+Z-VAD group was increased ( P<0.05), while the apoptosis rate and related protein ratios were significantly decreased (all P<0.05). Conclusion:SM-164 enhances the radiosensitivity of cervical cancer cells by inducing Casp activation.

Objective To investigate the effects of combined assessment of normal morphology sperm rate(MNS)with sperm DNA fragmentation index(DFI)on the outcomes of in vitro fertilization-embryo transfer(IVF-ET).Methods A retrospective analysis was conducted on the clinical data of 641 patients who underwent IVF-ET in our center from June 2020 to February 2023.According to MNS and DFI levels,the patients were divided into group A(MNS＜4％,DFI＜25％,403 cases),group B(MNS ≥4％,DFI ≥25％,9 ca-ses),group C(MNS≥4％,DFI＜25％,125 cases),group D(MNS＜4％,DFI≥25％,104 cases),group A1(MNS＜1％,DFI＜25％,106 cases)and group D1(MNS＜1％,DFI≥25％,60 cases).The general information,semen parameters,embryo develop-ment and pregnancy outcomes were compared among all the 6 groups.Results No statistically significant differences were found be-tween female and male age among these groups(P＞0.05).Significant differences were observed in sperm motility,progressive motili-ty,sperm concentration and fertilization rates(P＜0.05),but no differences of usable embryo rates,good-quality embryo rates,clinical pregnancy rates,early miscarriage rates,or live birth rates(P＞0.05)among the groups A,B,C,D,A1 and D1.Conclusion The decreased MNS and increased DFI in males were associated with reduced sperm motility and concentration,affecting fertilization rates in IVF-ET,but had no significant impact on clinical pregnancy outcomes.The combined assessment of MNS with DFI had limited pre-dictive value for IVF-ET pregnancy outcomes.

AIM:NLRP3 inflammasome was identified as the cellular machinery responsible for activation of inflammatory processes .The present study investigated whether the activation of NLRP 3 inflammasomes contributes to hyperhomocysteinemia ( HHcy)-induced in-flammation and atherosclerosis .METHODS:ApoE-/-mice were fed regular diet , high fat ( HF) diet or HF plus high methionine (HM) diet for 10 weeks.NLRP3 shRNA or scramble shRNA viral suspension was injected twice at the 2nd and the 6th weeks after HFHM treatment.The whole aortas and aortic root sections were stained with Oil Red O for atherosclerotic lesion .Plasma lipids, ho-mocysteine ( Hcy) , IL-1βand IL-18 levels were measured .We also examined the effect of Hcy on NLRP 3 inflammasomes activation in THP-1 differentiated macrophages in the presence or absence of NLRP 3 siRNA, caspase-1 inhibitor Z-WEHD-FMK, or antioxidant N-acetyl-L-cysteine ( NAC) .RESULTS:HFHM treatment induced HHcy in ApoE-/-mice.Increased plasma levels of IL-1βand IL-18, aggravated macrophage infiltration into atherosclerotic lesion , and accelerated development of atherosclerosis were detected in HHcy mice, which were associated with the activation of NLRP 3 inflammasomes.Silencing the NLRP3 gene significantly suppressed NLRP3 inflammasomes activation , reduced plasma levels of proinflammatory cytokines , attenuated macrophage infiltration , and improved HHcy-induced atherosclerosis .Moreover, we found that Hcy activated NLRP3 inflammasomes and promoted subsequent production of IL-1βand IL-18 in macrophages, which were blocked by NLRP3 gene silencing, Z-WEHD-FMK, or NAC.CONCLUSION:These data suggest that the activation of NLRP 3 inflammasomes contributes to HHcy-induced inflammation and atherosclerosis .Hcy activates NLRP3 inflammasomes in reactive oxygen species dependent pathway in macrophages .