ObjectiveTo investigate the effect of isoacteoside on the replication process of respiratory syncytial virus （RSV） and its underlying mechanism. MethodsRSV-infected HEp-2 cells （control group） were treated with isoacteoside （isoacteoside-treated group）. Real-time quantitative PCR （qPCR） was used to analyze the mRNA expression levels of the RSV fusion protein （F） gene and matrix protein （M） gene in both groups. Viral titers were determined by plaque assay to assess the inhibitory effect of isoacteoside on viral replication. The number of RSV inclusion bodies formed was determined using cell transfection and laser confocal imaging to evaluate the role of isoacteoside in the formation of inclusion bodies required for RSV replication. 18 seven-week-old female BALB/c mice were randomly divided into three groups： uninfected control group （n=6）， RSV-infected group （n=6）， and RSV-infected group treated with isoacteoside （n=6）. 5 days after infection， the mice were euthanized and lung tissues were collected for pathological analysis to assess the therapeutic effects of isoacteoside on RSV-induced lung tissue injury. ResultsCompared to the control group， the isoacteoside-treated group exhibited the reduced mRNA levels of the RSV F gene （t=17.13， P<0.001） and M gene （t=18.22， P<0.001）， as well as a decrease in viral titer （t=15.32， P<0.001）. The number of inclusion bodies was also significantly reduced （t=16.12， P<0.001）. In the RSV-infected mouse experiment， compared to the RSV-infected group， the isoacteoside-treated group showed a decrease in the levels of inflammatory cytokines IL-4 （t=14.76， P<0.01） and IL-6 （t=21.13， P<0.001） in lung tissues， as well as decreased mRNA levels of the viral load-associated genes F （t=19.52， P<0.001） and M （t=18.76， P<0.001） in lung tissues. Pathological damage to lung tissue caused by RSV infection was alleviated. ConclusionIsoacteoside inhibits RSV replication by interfering with the formation of RSV inclusion bodies and alleviates lung tissue injury induced by RSV infection in mice.

Objective : To explore the role of Golgi protein ACBD3 knockdown in ferroptosis. Methods : ACBD3 knockdown via shRNA , combined with treatment of ferroptosis inducer RSL3 , cell viability , iron levels , reactive oxygen species (ROS) levels , and lipid peroxide levels in different treatment groups were measured to comprehensively analyze the role of ACBD3 in ferroptosis. Results : Compared to control cells , the levels of ROS increased in ACBD3 knockdown cells. The total iron content was significantly reduced (P < 0. 001) , while the Fe2 + concentration in ACBD3 knockdown cells increased compared to control cells (P < 0. 001) . In the presence of ferroptosis inducer RSL3 , compared to control cells , cell viability significantly decreased ( P < 0. 001) , and lipid peroxide greatly increased in ACBD3 knockdown cells ( P < 0. 001) . Conclusion Golgi protein ACBD3 knockdown in mammalian cells changes iron homeostasis and promotes ferroptosis.

Objective : To explore the mechanism of Golgi phosphoprotein 3 (GOLPH3) regulating lysosomal biogenesis via mTORC1 signaling. Methods : GOLPH3 knockout (GOLPH3 KO) stable cell line was constructed by CRISPR Cas9. mTORC1 activity and the levels of TFEB and p ⁃TFEB in the control cells and GOLPH3 KO cells were compared through Western blot. Further, lysosome⁃associated membrane protein LAMP1 was labelled by the means of immunofluorescence and the number of lysosomes in the control cells and GOLPH3 KO cells was com⁃pared. Results : GOLPH3 KO suppressed mTORC1 activity significantly , decreased the cytoplasm level of p ⁃TFEB , increased the nuclear localization of active TFEB , and promoted the lysosome biogenesis. Conclusion Golgi protein GOLPH3 regulates lysosome biogenesis through mTORC1 signaling.