Bluetongue virus is an arbovirus that seriously harms ruminants such as sheep,this study aims to investigate the molecular mechanism of bluetongue virus infection and host cell interferon antiviral immune response.The study was conducted to characterize the mRNA expression of inter-feron pathway genes by real-time fluorescence quantitative PCR,as well as Western blot analysis of MDA5,TRAF3,RIG-Ⅰ,and TBK1 protein expression in BHK-21 cells induced by BTV with a multiplicity of infections(MOI)of 1 for 18,24,and 36 h.The results showed that the most pro-nounced changes in the expression of interferon signaling pathway genes were observed at 24 h of induction,the gene mRNA expression levels of the IFN-α,IFN-β,RIG-Ⅰ,TBK1,MDA5,VISA,and TRAF3 genes were upregulated.However,the mRNA expression levels of IKKε and TRAF6 genes were downregulated.At the protein level,MDA5 and TBK1 proteins were upregulated while RIG-1 and TRAF3 proteins were downregulated,which showed that BTV infection induces a typeⅠ interferon immune response in BHK-21 cells.This study lays the foundation for further exploring the antiviral immunity mechanism of IFN-Ⅰ signaling pathway regulatory genes in host cells infected with BTV infection.

Background and Aims:Kaempferol is a natural flavonoid compound that can regulate various processes and activities associated with cancer,such as the cell cycle,oxidative stress,apoptosis,proliferation,metastasis,and angiogenesis. Its potential for treating breast cancer has been validated in some studies,but its mechanism of action remains unclear. Therefore,this study was conducted to explore the potential targets and pathways of kaempferol in the treatment of breast cancer and establish a prognostic model.Methods:Using network pharmacology and bioinformatics approaches,the intersection targets of kaempferol for the treatment of breast cancer were obtained from databases such as HERB,GeneCards,STRING,PubChem,RCSB PDB,and TCGA. Protein interaction,GO analysis,and KEGG pathway enrichment analysis were performed,followed by molecular docking for validation. Prognostic-related genes were identified through LASSO-Cox regression analysis to establish a risk score model. The prognostic analysis for each gene,and the correlation between risk score and immune infiltration were analyzed. Results:Network pharmacology analysis identified 55 intersection targets. Protein interaction analysis revealed five potential key genes (Akt1,Bcl2,CASP3,ESR1,AR). GO analysis showed that kaempferol treatment of breast cancer involved 1604 biological process entries,20 cellular component entries,and 121 molecular function entries. KEGG pathway enrichment mainly included pathways related to chemical carcinogenesis,reactive oxygen species signaling,AGE-RAGE signaling in diabetic complications,TNF signaling,and IL-17 signaling. Molecular docking results indicated that kaempferol had a strong affinity for key targets,with the best binding effect observed with ESR1 (-9.1 kcal/mol). Ten prognostic genes (Bcl2,CYP1B1,DPP4,GSTM1,GSTM2,MMP1,NCOA2,NOS2,NR1I3,PTGS2) were obtained through LASSO-Cox regression analysis,and a risk score model was established. The AUC of this model for predicting breast cancer patient prognosis was greater than 0.5. Single-gene survival analysis indicated that higher expression of Bcl2 (HR=0.61,95％ CI=0.43-0.86,P=0.005),CYP1B1 (HR=0.68,95％ CI=0.49-0.94,P=0.022),GSTM1 (HR=0.68,95％ CI=0.47-0.98,P=0.037),GSTM2 (HR=0.64,95％ CI=0.46-0.90,P=0.010),and PTGS2 (HR=0.62,95％ CI=0.44-0.86,P=0.005) correlated with a higher overall survival (OS),while higher expression of MMP1 (HR=1.72,95％ CI=1.22-2.41,P=0.002),NCOA2 (HR=1.71,95％ CI=1.12-2.60,P=0.013),NOS2 (HR=1.67,95％ CI=1.20-2.32,P=0.002),and NR1I3 (HR=1.69,95％ CI=1.21-2.37,P=0.002) was associated with worse OS. The risk score was negatively correlated with the infiltration of T cells,CD8+T cells,myeloid dendritic cells,NK cells,and B cells,and positively correlated with monocyte/macrophage infiltration (all P<0.05).Conclusion:Kaempferol exerts therapeutic effects on breast cancer through multiple targets and pathways. The prognostic genes identified based on its related targets,along with the established prognostic model,can guide clinical treatment of breast cancer. The correlation between the prognostic model and immune infiltration provides direction for future experimental studies.

Background and Aims:Kaempferol is a natural flavonoid compound that can regulate various processes and activities associated with cancer,such as the cell cycle,oxidative stress,apoptosis,proliferation,metastasis,and angiogenesis. Its potential for treating breast cancer has been validated in some studies,but its mechanism of action remains unclear. Therefore,this study was conducted to explore the potential targets and pathways of kaempferol in the treatment of breast cancer and establish a prognostic model.Methods:Using network pharmacology and bioinformatics approaches,the intersection targets of kaempferol for the treatment of breast cancer were obtained from databases such as HERB,GeneCards,STRING,PubChem,RCSB PDB,and TCGA. Protein interaction,GO analysis,and KEGG pathway enrichment analysis were performed,followed by molecular docking for validation. Prognostic-related genes were identified through LASSO-Cox regression analysis to establish a risk score model. The prognostic analysis for each gene,and the correlation between risk score and immune infiltration were analyzed. Results:Network pharmacology analysis identified 55 intersection targets. Protein interaction analysis revealed five potential key genes (Akt1,Bcl2,CASP3,ESR1,AR). GO analysis showed that kaempferol treatment of breast cancer involved 1604 biological process entries,20 cellular component entries,and 121 molecular function entries. KEGG pathway enrichment mainly included pathways related to chemical carcinogenesis,reactive oxygen species signaling,AGE-RAGE signaling in diabetic complications,TNF signaling,and IL-17 signaling. Molecular docking results indicated that kaempferol had a strong affinity for key targets,with the best binding effect observed with ESR1 (-9.1 kcal/mol). Ten prognostic genes (Bcl2,CYP1B1,DPP4,GSTM1,GSTM2,MMP1,NCOA2,NOS2,NR1I3,PTGS2) were obtained through LASSO-Cox regression analysis,and a risk score model was established. The AUC of this model for predicting breast cancer patient prognosis was greater than 0.5. Single-gene survival analysis indicated that higher expression of Bcl2 (HR=0.61,95％ CI=0.43-0.86,P=0.005),CYP1B1 (HR=0.68,95％ CI=0.49-0.94,P=0.022),GSTM1 (HR=0.68,95％ CI=0.47-0.98,P=0.037),GSTM2 (HR=0.64,95％ CI=0.46-0.90,P=0.010),and PTGS2 (HR=0.62,95％ CI=0.44-0.86,P=0.005) correlated with a higher overall survival (OS),while higher expression of MMP1 (HR=1.72,95％ CI=1.22-2.41,P=0.002),NCOA2 (HR=1.71,95％ CI=1.12-2.60,P=0.013),NOS2 (HR=1.67,95％ CI=1.20-2.32,P=0.002),and NR1I3 (HR=1.69,95％ CI=1.21-2.37,P=0.002) was associated with worse OS. The risk score was negatively correlated with the infiltration of T cells,CD8+T cells,myeloid dendritic cells,NK cells,and B cells,and positively correlated with monocyte/macrophage infiltration (all P<0.05).Conclusion:Kaempferol exerts therapeutic effects on breast cancer through multiple targets and pathways. The prognostic genes identified based on its related targets,along with the established prognostic model,can guide clinical treatment of breast cancer. The correlation between the prognostic model and immune infiltration provides direction for future experimental studies.

Objective:To construct standard strains representing the main epidemic clades of HIV-1 in China, amplify the virus strains, and establish a seed lot.Methods:Six isolates of HIV-1 virus were identified and analyzed in genotype and phenotype, according to " interpretation for the social organization of the Standard strains of pathogenic microorganism- technical specifications for establishment of HIV strains". The isolates were amplified and cultivated to generate the secondary generation primary seed lot and the third generation working seed lot as frozen storage in liquid nitrogen. Results:Six HIV-1 standard strains were obtained, of which 3 strains are CRF_ 07BC (NRPC2.4.9003, NRPC2.4.9005, NRPC2.4.9006), 1 strain is CRF_ 01AE (NRPC2.4.9001), 1 strain is CRF_ 08BC (NRPC2.4.9002), and 1 strain is URF (NRPC2.4.9004). Phenotypic detection showed that all six strains are CCR5 tropics and Non syncytia inducing virus. TCID 50 were all greater than 1 × 10 5/ml, and concentrations of p24 antigen were all higher than 2 ng/ml. A primary seed lot with no less than 20 vials per strain and a working seed lot with no less than 50 vials per strain were constructed. The standard virus strains were used in evaluating antiviral drugs PEG2kC34, PEG5kC34, LP-19, and neutralizing antibody LSEVh LS-F. Conclusions:Six standard strains of HIV-1 virus covering the three main epidemic subtypes of HIV-1 in China have been obtained, and a storage of HIV-1 standard strain was constructed. It meets the need of the preservation of HIV-1 standard strains in China and provides support for drug and vaccine evaluation.

Objective:To assess the significance of counting the number of caudal vertebral ossification centers (OCN) below fetal terminal conus medullaris in the screening for closed spina bifida and tethered cord syndrome (TCS).Methods:The OCN was counted in 961 normal fetuses(normal group) between 17 and 41 gestational weeks and in 140 fetuses with closed spina bifida or tethered cord syndrome(abnormal group) from Jan.2013 to Dec.2020 in Affiliated Shenzhen Maternity & Child Healthcare Hospital, Southern Medical University, Women and Children′s Hospital, School of Medicine, Xiamen University and Maternity and Child Health Care of Guangxi Zhuang Autonomous Region. The OCN was counted in the dorsal mid-sagittal section of fetal caudal spine.The reliability and agreement test were evaluated by intraclass correlation coefficients in another 50 normal fetuses. The OCN was compared between two groups. ROC curve and the cut-off value were constructed and calculated.Results:In normal group, the N increased with the growing of gestational age.In the subgroup of 17-20 weeks, the OCN ranged from 5 to 7 in most fetuses. In the others subgroups, the OCN was equal to or greater than 6 in 99.9% cases and more than 6 in 97.1% cases. In abnormal group, OCN was less than 7 in 93.0% fetuses and less than 6 in 82.8% cases. There were statistical differences between the two groups except for the subgroup of 17-20 gestational weeks( P<0.05). With the cut-off value of 6.5, the specificity and sensitivity were 93.0% and 94.3% respectively for predicting the presence of closed spinal dysraphism or TCS. Conclusions:OCN is a simple way to evaluate the position of conus medullaris and to screen for the skin-covered spine dysraphism or TSC. OCN is more than 6 in most normal fetuses. Further evaluation of spine is required in fetuses with N less than or equal to 6.

Objective: The purpose of this review of COVID-19 related research is to deepen our understanding of SARS-CoV-2, which would be inspire new ideas for targeted drug development and vaccine design, and further empower the prevention and control COVID-19. Methods: Through literature research and data analysis, we explored the process and mechanism of epitranscriptomics modification to regulate the replication and infectivity of COVID-19. Results: Provide important ideas and technical support for the prevention and control of SARS-CoV-2 infections and emerging epidemic diseases. Conclusions Taking the new research direction of epitranscriptomics as the starting point, it is expected to open up new scientific research concepts and paradigms.

At the end of 2019, COVID-19 broke out and spread rapidly in our country. Several kinds of emergency plans for epidemic prevention and control were consulted and prepared in Fifth Medical Center, PLA General Hospital. Some specialists in Department of Surgery and Operation Room also made surgery procedure, infection prevention and control measures for COVID-19 patients, including medical staff management and training, pre-surgical evaluation and preparation of patients, procedures for transferring patients, intraoperative nursing cooperation, disinfection of instruments and environmental objects after surgery. The above measures have been continuously revised in the clinical work.

Objective: To understand the concentrations and source of SO42-,NO3-,Cl- and NH4+ in atmospheric PM2.5 in Zhoushan,and to provide reference for controlling PM2.5 and formulating effective environmental protection measures. Methods: Monitoring sites in new districts of Zhoushan were set up to continuously collect PM2.5 from 10th to 16th of each month and under the hazy weather during 2015-2016（AQI > 200）. The mass concentration of PM2.5 was measured by weighing method,and the concentrations of SO42-,NO3-,Cl- and NH4+ in PM2.5 components was detected by ion chromatography. Results: The average daily concentration of PM2.5 in Zhoushan from 2015 to 2016 was（40.91±27.39）μg/m3. The concentration of the four water-soluble non-metal ions in PM2.5 components was 3.56-103.03 μg/m3,with an average of（23.06±20.00）μg/m3,accounting for about 56.64% of PM2.5 contents. The average monthly concentration of SO42- was the highest[（10.35±6.48）μg/m3],while the average monthly concentration of Cl- was the lowest [（0.49±0.73）μg/m3]. The concentration of the four ions was the highest in winter[（37.56±27.74）μg/m3]and the lowest in summer[（12.32±5.88）μg/m3]. The differences between different seasons was statistically significant（P<0.05）. The highest concentration of NO3- occurred in winter,which was（14.48±13.28）μg/m3. The concentration ratio of NO3- to SO42- ranged from 0 to 2.58,with an average of 0.55. There were 28 days（14.74%）with the ratio greater than one,22 days of which was in winter. Conclusion The concentration of SO42- was the highest and Cl- was the lowest in atmospheric PM2.5 in Zhoushan. The highest concentration of the four ions occurred in winter. The concentration of NO3- in winter was higher than that of SO42-,suggesting that motor vehicle exhaust might be the main source of PM2.5 in winter.

BACKGROUND: Peanut allergy is an increasing problem in Singapore and strict avoidance is difficult as peanut is ubiquitous in Asian cuisine. OBJECTIVE: We aimed to assess the efficacy and safety of peanut oral immunotherapy (OIT) in children with obvious peanut allergy in Singapore. METHODS: This was an open-label study of peanut OIT in children living in Singapore, with 2 weekly dose escalation until final maintenance dose of 3,000 mg of peanut protein and a maintenance phase of 12 months. An oral food challenge was performed at 6 months to assess for desensitisation and at 4 weeks after discontinuation of OIT having completed 12 months of maintenance therapy to assess for possible sustained unresponsiveness. The adverse events were monitored using the symptom diaries. RESULTS: Nine subjects were started on OIT, with 7 managing to complete maintenance phase of therapy. Of these 7, all were able to tolerate at least 3,000 mg of peanut protein by 6 months of maintenance therapy, showing that the OIT was effective. Of these 7, 3 patients complied with the 4-week abstinence period after completion of OIT before another peanut challenge; 2 of the 3 subjects showed a significant decrease from the initial ability to tolerate 3,000 mg of peanut protein. Side effects were mainly gastrointestinal in nature and were more common during the updosing phase than the maintenance phase. No episodes of anaphylaxis were observed in this study. CONCLUSION: Peanut OIT seemed to be effective and safe in our cohort of Singaporean children.
Anaphylaxis ; Arachis ; Asian Continental Ancestry Group ; Child ; Cohort Studies ; Humans ; Hypersensitivity ; Immunotherapy ; Peanut Hypersensitivity ; Pilot Projects ; Probiotics ; Singapore

Objective To explore the influencing factors of blood concentration of tacrolimus in pediatric living donor liver transplant recipients and provide rationales for individualized administration of tacrolimus .Methods Trough concentrations (C0 ) , doses of tacrolimus , recipient age , gender , body weight ,donor and recipient CYP3A5 genotypes ,hematocrit (HCT ) and liver/kidney function related indicators at 3 ,5 ,7 ,14 days ,1 month , 2 months and 3 months post living donor liver transplantation were collected from a total of 100 pediatric recipients .Taking ratio of concentration to dose (C0 /D) as a dependent variable ,the influencing factors of blood concentration of tacrolimus were analyzed by multivariate stepwise regression .Results The influencing factors of blood tacrolimus concentration at 3d post-transplantation were recipient CYP3A5 genotyp , donor CYP3A5 genotype and weight of recipients . The major influencing factors at 5d post-transplantation were recipient & donor CYP3A5 genotypes , recipient weight and HCT . The major relevant factors at 7d posttransplantation were CYP3A5 of recipients ,age and HCT .The influencing factors at 14 days were the same as those at 2 months ,i .e .CYP3A5 genotype and weight of recipients .At 1 month the major influencing factors were weight of recipients ,CYP3A5 of recipients and alkaline phosphatase (ALP) ; CYP3A5 genotype and weight of recipients at 3 months . Further study on CYP3A5 genotype of donors and recipients , the C0 /D ratio of CYP3A5 genotype non-expression group was significantly higher than that of expression group in recipients and C0 /D ratio of donor CYP3A5 genotype nonexpression group was significantly higher than that of expression group .Conclusions The influencing factors of concentration of tacrolimusvary at different timepoints after liver transplantation . Paying close attention to the changes of CYP3A5 genotype , weight of recipients and related biochemical indexes and considering various influencing factors facilitate individualized dosing for improving the prognosis of pediatric recipients .