Microglia, the resident immune cells in the central nervous system (CNS), rapidly transition from a resting to an active state in the acute phase of ischemic brain injury. This active state mediates a pro-inflammatory response that can exacerbate the injury. Targeting the pro-inflammatory response of microglia in the semi-dark band during this acute phase may effectively reduce brain injury. Shionone (SH), an active ingredient extracted from the dried roots and rhizomes of the genus Aster (Asteraceae), has been reported to regulate the inflammatory response of macrophages in sepsis-induced acute lung injury. However, its function in post-stroke neuroinflammation, particularly microglia-mediated neuroinflammation, remains uninvestigated. This study found that SH significantly inhibited lipopolysaccharide (LPS)-induced elevation of inflammatory cytokines, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS), in microglia in vitro. Furthermore, the results demonstrated that SH alleviated infarct volume and improved behavioral performance in middle cerebral artery occlusion (MCAO) mice, which may be attributed to the inhibition of the microglial inflammatory response induced by SH treatment. Mechanistically, SH potently inhibited the phosphorylation of serine-threonine protein kinase B (AKT), mammalian target of rapamycin (mTOR), and signal transducer and activator of transcription 3 (STAT3). These findings suggest that SH may be a potential therapeutic agent for relieving ischemic stroke (IS) by alleviating microglia-associated neuroinflammation.
Animals ; Microglia/immunology* ; Mice ; Male ; Mice, Inbred C57BL ; Brain Ischemia/immunology* ; Neuroinflammatory Diseases/drug therapy* ; Neuroprotective Agents/administration & dosage* ; Interleukin-1beta/genetics* ; STAT3 Transcription Factor/genetics* ; TOR Serine-Threonine Kinases/genetics* ; Tumor Necrosis Factor-alpha/genetics* ; Proto-Oncogene Proteins c-akt/immunology* ; Nitric Oxide Synthase Type II/genetics* ; Lipopolysaccharides

ObjectiveThe human angiotensin converting enzyme2 （hACE2） transgenic mouse model was used to clarify the antiviral efficacy of BD-77 against a novel coronavirus SARS-CoV-2 and explore the action mechanism of BD-77 against SARS-CoV-2. MethodSARS-CoV-2 Omicron and Delta variant strains-infected VeroE6 cell models were established and administered with BD-77 to observe the antiviral effect of BD-77 in vitro. A kit was used to detect the effect of BD-77 in vitro on the binding of spike S protein of SARS-CoV-2 virus （Delta/Omicron） to angiotensin converting enzyme2 （ACE2）. Chromatography was adopted to detect the binding of BD-77 to the S protein and N protein of the novel coronavirus. hACE2 transgenic C57BL/6 mice were divided into a blank control group， SARS-CoV-2 infection group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1， with eight mice in each group. The pneumonia model of SARS-CoV-2-infected hACE2 transgenic mice was built to observe the survival of the mice， detect the virus titer of the lung tissue of the mice， and observe the lesions in the lung tissue. ResultBD-77 had a certain inhibitory effect on Omicron and Delta variant strains in vitro， with median inhibitory concentration （IC₅₀） of 526.3 mg·L^-1 and 653.0 mg·L^-1， respectively. BD-77 had no significant inhibitory effect on the binding of the S protein of WT， Omicron， and Delta variant strains of SARS-CoV-2 to ACE2 and had no binding effect with the S protein and N protein of the novel coronavirus. No mice in the blank group died， while the mortality rate of SARS-CoV-2-infected mice was 75%. There was a large amount of virus replication in the lung tissue of the mice and large areas of inflammatory infiltration in the lung tissue and interstitium. Compared with the model group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1 could reduce the mortality of mice， significantly lower the virus titer in the lung tissue of mice （P<0.05）， and improve lung lesions. ConclusionBD-77 demonstrated significant inhibitory effects against SARS-CoV-2 virus in vitro and in vivo. However， its mechanism of action did not involve direct inhibition of the virus itself or intervention in the virus-host binding process. This finding suggests that the mechanism of action of BD-77 needs to be thoroughly investigated and elucidated by further experiments.

ObjectiveTo observe the therapeutic effect of BD-77 by nebulized inhalation on animal models of various respiratory viral infections and investigate the mechanism of broad-spectrum antiviral action of BD-77 using proteomics. MethodThe influenza virus H1N1/FM1 experiment used ICR mice and divided them into a normal group， model group， Tamiflu group， and BD-77 groups of 75 and 37.5 g·L^-1 for inhalation of 20 min and 25 min. Human coronavirus 229E and OC43 experiment divided the BALB/c mice into a normal group， model group， chloroquine phosphate group， and BD-77 groups of 75， 37.5， 18.75， and 9.375 g·L^-1， with 10 mice in each group. Influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection-induced pneumonia models were used to detect mouse lung index， and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the viral load in lung tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect related inflammatory factors in lung tissue， and proteomics analysis was performed on the lung tissue of OC43-infected mice. ResultCompared with that in the normal group， the lung index of mice in each infection group was significantly increased （P<0.01）， and viral nucleic acid could be detected in the lung tissue of mice infected with human coronaviruses 229E and OC43. The levels of interleukin-6 （IL-6）， IL-10， and tumor necrosis factor-α （TNF-α） in the lung tissue of mice infected with human coronavirus 229E were all significantly increased （P<0.01）. BD-77 could significantly reduce the lung index of mice infected with influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 （P<0.05， P<0.01）， cut down the viral load in the lungs of mice infected with human coronaviruses 229E and OC43 （P<0.01）， and lower the contents of IL-6， IL-10， and TNF-α in the lung tissue of mice infected with human coronavirus 229E （P<0.01）. Proteomics analysis of the lung tissue of OC43-infected mice showed that BD-77 regulated the AMPK signaling pathway， TNF signaling pathway， NOD-like signaling pathway， IL-17 signaling pathway， Forkhead box protein O （FoxO） signaling pathway， transforming growth factor-β （TGF-β） signaling pathway， and other signaling pathways. ConclusionNebulized inhalation of BD-77 is effective in treating pneumonia caused by influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection in mice and may exert its antiviral effects by regulating the balance of cellular metabolism， enhancing the immune function of the host， and attenuating inflammatory responses.

Objective : To explore the role and molecular mechanism of Supervillin in zebrafish embryo development. Methods : Clustal Omega and DNAman software were used to analyze and compare the homology of amino acid sequences of Svil and SVIL in zebrafish and human. The expression pattern of Svila (subtype of Svil protein in zebrafish) during early embryonic development was analyzed by RT⁃PCR and in situ hybridization. Svila expression in zebrafish was inhibited by injecting morphinos ( MO) , and morphological changes of embryos were observed. The expression and nuclear localization of β ⁃catenin protein were detected by Western blot , and the expression of Wnt/β⁃catenin target gene was detected by RT⁃PCR. Results : During the early embryonic development of zebrafish , Svila was maternally expressed and showed an upward trend with the development process. The expression of Svila was reduced by MO , and the development of zebrafish embryos was distorted and the body axis was bent , which might be related to the blocked movement of concentrated extension of embryos. Further studies showed that Svila expression affected β⁃catenin nuclear transport and Wnt/β⁃catenin signaling pathway activation. Conclusion Svila regulates the concentrated extension movement of zebrafish embryos by activating the Wnt/β⁃catenin signaling pathway.

Objective:To evaluate arthroscopy-assisted tibial bone tunnel fixation combined with double-row anchors in the treatment of tibial eminence fracture.Methods:The 23 patients were retrospectively analyzed who had been treated at Department of Orthopedics, Beiing Shunyi District Hospital for tibial eminence fractures by arthroscopy-assisted tibial bone tunnel fixation combined with double-row anchors from October 2015 to December 2019. They were 15 males and 8 females, aged from 12 to 55 years (average, 35.5 years). All the injuries were unilateral (14 right and 9 left sides). According to the modified Meyers-McKeever classification, 2 cases belonged to type Ⅱ, 18 cases to type Ⅲ and 3 cases to type Ⅳ. Range of motion of the knee, Lysholm and International Knee Documentation Committee (IKDC) scores were observed before surgery, 1 month and 12 months after surgery. The anterior tibial slope angle (ATSA) on CT was measured preoperatively and 1 month after surgery for evaluation of fracture reduction.Results:All the 23 patients were followed up for an average of 23 months (from 12 to 52 months). Postoperatively, limited knee movement was observed in 2 patients and non-anatomic reduction in one patient. At 1 month and 12 months after operation, the Lysholm scores (61.4 ± 3.5 and 90.4 ± 4.3) and IKDC scores (69.6 ± 4.2 and 88.5 ± 3.0) were significantly improved compared with the preoperative values (45.4 ± 6.8 and 49.6 ± 3.9, respectively) ( P<0.05). ATSA was significantly restored from preoperative 4.2° ± 5.7° to -11.7° ± 2.9° at 1 month after operation ( P<0.05). Conclusion:In the treatment of tibial eminence fracture, arthroscopy-assisted tibial bone tunnel fixation combined with double-row anchors can achieve anatomical reduction and firm fixation, leading to satisfactory surgical outcomes.

Objective:To explore the reproductive health status of the female street cleaners in Chaoyang district of Beijing and its influencing factors.Methods:In July 2018, a total of 647 questionnaires were sent out to female road cleaning and sanitation workers in the whole district, 613 of which were valid, with an effective rate of 94.7%. Frequency or percentage (%) is used for statistical description of counting data, and mean standard deviation is used for measurement data. The influencing factors were analyzed by 2 test.Results:The average age of 613 female road sweepers in Chaoyang District of Beijing was 42.01 (SD=6.69) years old, including 535 married female workers (87.28%) , 356 middle school and below educated (58.08%) , 292 non Beijing registered (47.63%) , accounting for (/613) , (/613) female workers working hours >8 hours per day 110 (17.94%) , weekly rest <2 days 341 (55.63%) . 144 (23.49%) women workers did not have regular gynecological examination, 119 (19.41%) had reproductive tract infections, 177 (28.87%) had abnormal menstruation in recent 6 months. Drinking, sexual behavior outside marriage, night shift and job satisfaction were all the influencing factors ( P<0.05) . The increasing age, working years, drinking, household registration in other places and the decreasing satisfaction of women workers＇ rights and interests protection were all the influencing factors ( P<0.05) . Conclusion:The reproductive health status of female road sweepers in Chaoyang District of Beijing is not optimistic.

Objective:To explore the reproductive health status of the female street cleaners in Chaoyang district of Beijing and its influencing factors.Methods:In July 2018, a total of 647 questionnaires were sent out to female road cleaning and sanitation workers in the whole district, 613 of which were valid, with an effective rate of 94.7%. Frequency or percentage (%) is used for statistical description of counting data, and mean standard deviation is used for measurement data. The influencing factors were analyzed by 2 test.Results:The average age of 613 female road sweepers in Chaoyang District of Beijing was 42.01 (SD=6.69) years old, including 535 married female workers (87.28%) , 356 middle school and below educated (58.08%) , 292 non Beijing registered (47.63%) , accounting for (/613) , (/613) female workers working hours >8 hours per day 110 (17.94%) , weekly rest <2 days 341 (55.63%) . 144 (23.49%) women workers did not have regular gynecological examination, 119 (19.41%) had reproductive tract infections, 177 (28.87%) had abnormal menstruation in recent 6 months. Drinking, sexual behavior outside marriage, night shift and job satisfaction were all the influencing factors ( P<0.05) . The increasing age, working years, drinking, household registration in other places and the decreasing satisfaction of women workers＇ rights and interests protection were all the influencing factors ( P<0.05) . Conclusion:The reproductive health status of female road sweepers in Chaoyang District of Beijing is not optimistic.

Objective To verify the antibacterial activity of the conserved domain derived from the novel human antimicrobial peptide-hGlyrichin .Methods Bioinformative analysis was performed and two peptides derived from hGlyrichin were synthesized which contained the conserved domain .Results Analysis of antimicrobial activities showed that these two peptides exhibited strong antibacterial activity which was inversely proportional to the length of the peptide within an eligible range.Despite the effective inhibition and killing of bacteria , the synthetic peptide segments had no hemolytic effect on human red blood cells .Conclusion These results indicate that a conserved domain exists in hGlyrichin , and that the peptides which contain this domain have strong antibacterial activity but are not toxic to human somatic cells .

Objective To observe the effects of hyperbaric oxygen (HBO) on learning and memory ability and on the level of myelin basic protein (MBP) in the brain tissue of rats with delayed neurological sequelae (DNS) after acute carbon monoxide poisoning (COP).Methods Forty-eight male SD rats had their cognitive performance assessed with the Morris water maze.After basic training with the Morris water maze and screening,the rats were randomly divided intoanormalcontrol (NC) group (n=11),a COP group (n=17) and an HBO group (n=17).Pure CO gas was injected intraperitoneally to establish acute COP in the latter two groups.The NC group received a similar injection of air.The HBO group was given HBO therapy.The rats in each group were tested for changes in their learning and memory abilities using the Morris water maze.On the 21 st day after the treatment,paraffin tissue sections of the rats' brains were subjected to immunohistochemical (IHC) examination and Western blotting (WB) was used to detect any expression of MBP in the brain tissue.Results After 21 days,morbidity among the COP group was 64.3％,while it was 26.7％ in the HBO group,a significant difference.The average maze escape latency in the COP group was significantly longer than in the HBO group.IHC staining and Western blotting showed that MBP in the hippocampal tissue of the COP group was significantly lower than that in the HBO group.In gray scale comparisons of the rats' brain tissue,that from the NC group was significantly better than that from the COP and HBO groups,but that from the HBO group was significantly better than that from the COP group.Conclusion HBO can effectively reduce DNS after acute COP,mitigate the severity of DNS,reduce demyelination of brain tissue and thus play an important role in protecting brain cells.

OBJECTIVETo study the influence of Aconiti Lateralis Radix Praeparata on asthenia cold syndrome rats with whole genome gene expression of liver by gene chip technique.

METHODThe asthenia cold syndrome rat models were established by administering traditional Chinese medicine raw Gypsum Fibrosum, Gentianae Radix, Phellodendri Chinensis Cortex and Anemarrhenae Rhizoma. After treated with Aconiti Lateralis Radix Praeparata, the rats' liver gene expressions were detected using gene chip. Differential expression genes were screened for gene function annotation, and some genes were selected to check the accuracy of the results by RT-PCR.

RESULTCompared with the asthenia cold model group, the asthenia cold treatment group showed 212 differential expression genes, mainly involving function of immune response and oxidoreductase activity.

CONCLUSIONAconiti Lateralis Radix Praeparata is proved to have an effect on up-regulating immune response-related genes and oxidizing oxidoreductase activity-related genes of asthenia cold syndrome rats and may be a molecular mechanism for classical warm-nature medicine Aconiti Lateralis Radix Praeparata in warming meridians and dissipating cold.

Aconitum ; chemistry ; Animals ; Asthenia ; drug therapy ; genetics ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Profiling ; Genomics ; Liver ; drug effects ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Wistar