ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， network pharmacology， molecular docking and cell experiments， the active ingredients， possible targets and molecular mechanisms of Baoxin granules（BXG） regulating ferroptosis in the treatment of heart failure（HF） were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components， the potential targets of BXG were predicted， and the HF-related targets and related genes of ferroptosis were retrieved at the same time， and the intersecting targets were obtained by Venn diagram. In addition， the protein-protein interaction（PPI） network and the component-target network were constructed， and the core components and core targets were obtained by topological analysis. Then Gene Ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis were performed on the core targets， and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model， and the protective effect of BXG on cells was investigated by detecting cell viability， cell survival rate and reactive oxygen species（ROS） level. The protein expression levels of signal transducer and activator of transcription 3（STAT3）， phosphorylation（p）-STAT3 and glutathione peroxidase 4（GPX4） were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified， and network pharmacology obtained 27 potential targets of BXG for the treatment of HF， as well as 139 signaling pathways. BXG may act on core targets such as STAT3， tumor protein p53（TP53）， epidermal growth factor receptor（EGFR）， JUN and prostaglandin-endoperoxide synthase 2（PTGS2） through core components such as glabrolide and limonin， which in turn intervene in lipid and atherosclerosis， phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）， endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group， the cell viability and survival rate of the model group were significantly decreased， the level of ROS was significantly increased（P<0.01）， the expression levels of STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 proteins were significantly decreased（P<0.05， P<0.01）. Compared with the model group， the cell viability and survival rate of the BXG group were significantly increased， the ROS level was significantly decreased（P<0.01）， the STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 protein levels were significantly increased（P<0.05， P<0.01）. ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4， thus exerting a therapeutic effect on HF， and flavonoids may be the key components of this role.

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， network pharmacology， molecular docking and cell experiments， the active ingredients， possible targets and molecular mechanisms of Baoxin granules（BXG） regulating ferroptosis in the treatment of heart failure（HF） were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components， the potential targets of BXG were predicted， and the HF-related targets and related genes of ferroptosis were retrieved at the same time， and the intersecting targets were obtained by Venn diagram. In addition， the protein-protein interaction（PPI） network and the component-target network were constructed， and the core components and core targets were obtained by topological analysis. Then Gene Ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis were performed on the core targets， and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model， and the protective effect of BXG on cells was investigated by detecting cell viability， cell survival rate and reactive oxygen species（ROS） level. The protein expression levels of signal transducer and activator of transcription 3（STAT3）， phosphorylation（p）-STAT3 and glutathione peroxidase 4（GPX4） were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified， and network pharmacology obtained 27 potential targets of BXG for the treatment of HF， as well as 139 signaling pathways. BXG may act on core targets such as STAT3， tumor protein p53（TP53）， epidermal growth factor receptor（EGFR）， JUN and prostaglandin-endoperoxide synthase 2（PTGS2） through core components such as glabrolide and limonin， which in turn intervene in lipid and atherosclerosis， phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）， endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group， the cell viability and survival rate of the model group were significantly decreased， the level of ROS was significantly increased（P<0.01）， the expression levels of STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 proteins were significantly decreased（P<0.05， P<0.01）. Compared with the model group， the cell viability and survival rate of the BXG group were significantly increased， the ROS level was significantly decreased（P<0.01）， the STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 protein levels were significantly increased（P<0.05， P<0.01）. ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4， thus exerting a therapeutic effect on HF， and flavonoids may be the key components of this role.

Objective:To investigate the effects of maternal antibiotics during the pre-natal period on the develop-ment of microglia and anxiety-like behaviors in offspring.Methods:Female SD rats were randomly divided into a maternal control group(CON-M)and a maternal antibiotic group(ABX-M),with 5 rats in each group.The antibiotic group received oral administration of compound antibiotics for 7 days before conception,and the control group received oral administration of the same dose of 0.9％saline during the same period.Subsequently,the two groups of female rats completed the pregnancy and delivery process at the same time,and their male offspring were retained and marked as the offspring control group(CON-O)and the offspring antibiotic group(ABX-O),respectively.The anxiety-like behaviors of offspring rats at 4 weeks and 8 weeks of age were evaluated by open field test and elevated plus maze test,respectively.The number and morphology of microglia in the hippocampus(HIP),cingulate gyrus(CG)and basolater-al nucleus of amygdale(BLA)of the brain of each group of offspring rats were detected by laser confocal microscopy im-aging.Results:The body weight and number of offspring were reduced in the ABX-M group.The 4-week-old ABX-O group showed an increase in anxiety-like behaviors,accompanied by an increase in the number and immature phenotype of microglia in the hippocampus,cingulate gyrus,and amygdala.Compared with the 8-week-old CON-O group,the 8-week-old ABX-O group showed obvious anxiety-like behaviors,and the number and immature phenotype of microglia in the hippocampus,cingulate gyrus,and amygdala of the brain continued to increase.Conclusion:Maternal antibiotic exposure during the pre-pregnancy period significantly affects the number and maturation of microglia in offspring,lead-ing to anxiety-like behaviors,and its effects can persist into adulthood.

Objective:To investigate the effects of maternal antibiotics during the pre-natal period on the develop-ment of microglia and anxiety-like behaviors in offspring.Methods:Female SD rats were randomly divided into a maternal control group(CON-M)and a maternal antibiotic group(ABX-M),with 5 rats in each group.The antibiotic group received oral administration of compound antibiotics for 7 days before conception,and the control group received oral administration of the same dose of 0.9％saline during the same period.Subsequently,the two groups of female rats completed the pregnancy and delivery process at the same time,and their male offspring were retained and marked as the offspring control group(CON-O)and the offspring antibiotic group(ABX-O),respectively.The anxiety-like behaviors of offspring rats at 4 weeks and 8 weeks of age were evaluated by open field test and elevated plus maze test,respectively.The number and morphology of microglia in the hippocampus(HIP),cingulate gyrus(CG)and basolater-al nucleus of amygdale(BLA)of the brain of each group of offspring rats were detected by laser confocal microscopy im-aging.Results:The body weight and number of offspring were reduced in the ABX-M group.The 4-week-old ABX-O group showed an increase in anxiety-like behaviors,accompanied by an increase in the number and immature phenotype of microglia in the hippocampus,cingulate gyrus,and amygdala.Compared with the 8-week-old CON-O group,the 8-week-old ABX-O group showed obvious anxiety-like behaviors,and the number and immature phenotype of microglia in the hippocampus,cingulate gyrus,and amygdala of the brain continued to increase.Conclusion:Maternal antibiotic exposure during the pre-pregnancy period significantly affects the number and maturation of microglia in offspring,lead-ing to anxiety-like behaviors,and its effects can persist into adulthood.

ObjectiveTo investigate the effects of thyme herbal tea （BLX） on the proliferation of human coronavirus OC43 （HCoV-OC43） in vitro and in vivo. MethodThe chemical composition of BLX was analyzed by UPLC-MS. The cytotoxicity of BLX in HRT-18 cells and the effect of BLX treatment on the proliferation of HCoV-OC43 in cells were analyzed. Copies of viral gene were detected by real-time PCR. The effect of BLX treatment on the life cycle of HCoV-OC43 was detected by time-of-addition assay. The maximum tolerated dose of BLX and the influences of BLX on the body weight and survival time of suckling mice infected with HCoV-OC43 were determined. The expression of viral protein in the brain and lung tissue was analyzed by immunohistochemistry. ResultThere were 11 chemical components identified in BLX by UPLC-MS. BLX showed the 50% cytotoxic concentration （CC₅₀） of （13 859.56±319） mg·L^-1， the median inhibitory concentration （IC₅₀） of （1 439.09±200） mg·L^-1， and the selection index of 8.26-11.44 for HCoV-OC43 in HRT-18 cells. Compared with the cells infected with HCoV-OC43， BLX at the concentrations of 1 500， 1 000， 500 mg·L^-1 inhibited the proliferation of this virus （P<0.05， P<0.01）. BLX exhibited antiviral effect in the early stage of virus infection， and the inhibition role in the attachment stage was more significant than that in the entry stage （P<0.05）. In the suckling mice infected with HCoV-OC43， BLX at 1200 and 600 mg·kg^-1·d^-1 alleviated the symptoms， prolonged the survival period， reduced the death rate， and down-regulated the mRNA level of nucleocapsid protein in the mice. Moreover， BLX at 1 200 mg·kg^-1·d^-1 down-regulated the expression of nucleocapsid protein in the brain （P<0.01） and the lung （P<0.01）. ConclusionBLX contained multiple antiviral ingredients. It inhibited the proliferation of HCoV-OC43 both in vitro and in vivo by interference with viral attachment. This study provides theoretical reference for the treatment of acute respiratory tract infection with HCoV-OC43 and for further development and application of BLX.

Objective:To study the course and mechanism of the immune response to SARS virus. Methods:The recombinant SARS virus S1 subunit was expressed in E. Coli according to the results of bioinformatics analysis. After purification, the recombinant S1 protein was identified by 6 serum samples of recovered SARS patients and 6 serum samples of health donors, which were collected before out-break of SARS. Results:Sequencing analysis confirmed that the recombinant protein has the same sequence of natural SARS virus S1 subunit. The recombinant S1 protein could react with all the samples from recovered SARS patients but not the control samples from healthy donors according to the results of Western blot. Conclusion:The recombinant SARS virus S1 subunit may provide a good tool for the research of immune response to SARS virus and the producing of recombinant vaccine to prevent people from SARS.