ObjectiveTo explore the improving effect of Huangqi Chifengtang（HCT） on atherosclerosis（AS）， and elucidate its mechanism in relation to adenosine monophosphate-activated protein kinase（AMPK）/peroxisome proliferator-activated receptor α（PPARα） signaling pathway and nucleotide-binding oligomerization domain（NOD）-like receptor thermal protein domain associated protein 3（NLRP3） inflammasome. MethodsEight C57BL/6J mice were set as the normal group， and 32 ApoE^-/- mice were randomly divided into the model group， the positive drug group（atorvastatin， 5 mg·kg^-1·d^-1）， HCT low- and high-dose groups（1.95， 3.90 g·kg^-1·d^-1）. ApoE^-/- mice were fed with high-fat and high-cholesterol feed to establish an AS mouse model. After modeling， they were orally administered corresponding dose of drugs for 28 days， while the normal and model groups received an equal volume of physiological saline via oral gavage. Hematoxylin-eosin（HE） staining was used to observe the pathological status of the aorta and liver in mice， Biochemical testing and enzyme-linked immunosorbent assay（ELISA） were used to detect the levels of total cholesterol（TC）， triglycerides（TG）， low-density lipoprotein cholesterol（LDL-C）， alanine aminotransferase（ALT）， aspartate aminotransferase（AST）， C-reactive protein（CRP）， interleukin（IL）-1β， IL-18 in the serum， as well as superoxide dismutase（SOD）， malondialdehyde（MDA）， and reduced glutathione（GSH） in the liver. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was used to measure the mRNA expression levels of NLRP3， apoptosis-associated speck-like protein（ASC）， cysteinyl aspartate specific proteinase-1（Caspase-1）， Toll-like receptor 4（TLR4） in the aorta， and fatty acid synthase（FAS）， stearoyl-CoA desaturase 1（SCD1）， PPARα， and carnitine palmitoyltransferase 1A（CPT1A） in the liver. Immunohistochemistry was used to determine the protein expressions of NLRP3， Caspase-1， and ASC in the aorta， and Western blot was used to measure the protein expressions of AMPK， p-AMPK， sterol regulatory element binding protein-1c（SREBP-1c）， CPT1A， and FAS in the liver. ResultsCompared with the normal group， the model group showed a significant increase in lipid plaque deposition in the aorta and lipid accumulation in the liver， the levels of TC， TG， LDL-C， AST， ALT， IL-1β， IL-18 and CRP in the serum were significantly increased（P<0.01）， and the mRNA and protein expressions of aortic TLR4， NLRP3， Caspase-1 and ASC were significantly upregulated（P<0.01）. The levels of SOD and GSH in the liver were significantly reduced， while the level of MDA was significantly increased（P<0.01）. The mRNA expressions of FAS and SCD1 in the liver were significantly downregulated， while the mRNA expressions of PPARα and CPT1A were significantly upregulated. The protein expressions of p-AMPK/AMPK and CPT1A in the liver were significantly reduced， while the expressions of SREBP-1c and FAS proteins were significantly increased（P<0.01）. Compared with the model group， the low- and high-dose HCT groups showed significant improvements in aortic plaques and hepatic lipid deposition. The levels of TC， LDL-C， AST， IL-1β and IL-18 in the serum of the low-dose HCT group， as well as TC， TG， LDL-C， AST， ALT， IL-1β， IL-18 and CRP in the serum of the high-dose HCT group， were significantly reduced（P<0.01）. The mRNA expressions of TLR4， NLRP3 and Caspase-1 in the aorta of the low-dose HCT group， as well as TLR4， NLRP3， Caspase-1 and ASC in the aorta of the high-dose HCT group， were significantly downregulated（P<0.01）. The protein expressions of Caspase-1 and ASC in the aorta of the low-dose HCT group， as well as NLRP3， Caspase-1 and ASC in the high-dose HCT group， were significantly downregulated（P<0.01）. The levels of SOD and GSH in the liver of the low- and high-dose HCT groups were significantly increased， while the level of MDA in the high-dose HCT group was significantly decreased（P<0.05， P<0.01）. In the HCT-treated group， the mRNA expressions of FAS and SCD1 in the liver were significantly upregulated， while the mRNA expressions of PPARα and CPT1A were significantly downregulated， the protein expressions of p-AMPK/AMPK and CPT1A in the liver were significantly increased， while the protein expressions of SREBP-1c and FAS were significantly decreased（P<0.05， P<0.01）. ConclusionHCT can improve lipid metabolism by activating the AMPK/PPARα pathway and inhibit NLRP3 inflammasome-mediated inflammatory responses， thereby reducing hepatic lipid deposition and AS plaque formation.

Objective To analyze the monitoring data of cardio-cerebrovascular diseases prevention and control system in Yichang in 2022, and to provide data support and experience for the precise prevention and treatment of cardio-cerebrovascular diseases. Methods Acute cardiovascular and cerebrovascular event data were collected from the Yichang Cardio-cerebrovascular Events Monitoring System from January 1, 2022 to December 31, 2022. Descriptive analysis was conducted for the data collected. Statistical analysis was performed using SPSS 20.0 software, and a chi-square test was used to analyze the count data. Results A total of 37,217 cases of cardio-cerebrovascular events were monitored in Yichang in 2022. The crude incidence and the standardized incidence were 983.84/100,000 and 541.55/100,000, respectively. The incidence in males was higher than females (554.93/100,000 vs 428.91/100,000，χ² =464.52，P＜0.05). The top three diseases were cerebral infarction, acute myocardial infarction, and cerebral hemorrhage. The incidence of events increased with age, and 79.80% of the cases were over 60 years old. The main onset time was from May to August. Conclusion The use of the cardio-cerebrovascular events monitoring system in Yichang and the implementation of ^“mandatory reporting card^” monitoring can timely obtain the epidemic characteristics of the diseases, provide support for the precise formulation of prevention and control strategies and measures, reduce underreporting rates, and improve the monitoring system, which is worthy of reference and promotion.

AIM To investigate the effects of Huazhuo Jiedu Shugan Formula(HJSGF)on improving learning and memory impairment in a rat model of vascular dementia(VD)via SIRT1/PGC-1α/PPARγ pathway.METHODS The SD rats were randomly divided into the sham control group,the model group,the donepezil group(0.5 mg/kg),and the low-,medium-and high-dose HJSGF groups(2.7,5.4,10.8 g/kg),with 10 rats in each group.The VD rat models established by bilateral common carotid artery permanent ligation(2-VO)had their neurological behavior assessment using the Longa5-point scale,and their modeling success confirmed by the Morris water maze test and their 3-week corresponding dosing of drugs by gavage afterward.After the drug administration,the rats had their spatial memory ability tested through behavioral experiments;their serum levels of IL-18 and IL-1β measured by ELISA;their histopathological changes and neuronal morphology in the hippocampal CA1 region observed by HE staining and Nissl staining;and their hippocampal protein expressions of SIRT1,PGC-1α and PPARγ detected by immunohistochemistry and Western blot.RESULTS Compared with the sham control group,the model group showed prolonged escape latency(P＜0.01);decreased platform crossing times and target quadrant residence time(P＜0.01);disorganized arrangement of hippocampal CA1 neurons,nuclear condensation,reduced Nissl bodies,increased secretion and protein expressions of IL-1β and IL-18(P＜0.01);and reduced hippocampal protein expressions of SIRT1,PGC-1α and PPARγ(P＜0.01).Compared with the model group,the groups intervened with donepezil or HJSGF showed shortened escape latency(P＜0.05,P＜0.01);increased platform crossing times and target quadrant residence time(P＜0.05,P＜0.01);alleviated damage of the hippocampal CA1 region,reduced secretion and protein expressions of IL-1β and IL-18(P＜0.05,P＜0.01);and elevated hippocampal protein expressions of SIRT1,PGC-1α and PPARγ(P＜0.05,P＜0.01).CONCLUSION HJSGF may alleviate the inflammatory responses in VD rats and therefore improve their learning and memory impairment by activating the SIRT1/PGC-1α/PPARγ signaling pathway.

BACKGROUND:The pathogenesis of steroid-induced osteonecrosis of the femoral head is not yet fully understood.A deep comprehension of the pathology and molecular mechanisms of steroid-induced osteonecrosis of the femoral head,as well as the search for diagnostic markers with high specificity and sensitivity,are crucial for the prevention and treatment of this condition.OBJECTIVE:To identify immune diagnostic markers for steroid-induced osteonecrosis of the femoral head and predict potential drug targets through drug enrichment analysis and molecular docking techniques.METHODS:The study utilized gene expression profile data(GSE123568 and GSE74089)from the GEO databases(a public gene expression database built by the U.S.National Center for Biotechnology Information).R software was used for data normalization and differential gene screening,followed by weighted gene co-expression network analysis(WGCNA)to identify disease-related genes.Immune-related genes were obtained from the GeneCards database and intersected with the differential genes and WGCNA gene sets to select immune-related genes for steroid-induced osteonecrosis of the femoral head.Mendelian randomization was used to validate the potential causal relationship between these immune-related genes and steroid-induced osteonecrosis of the femoral head.Gene Set Enrichment Analysis was conducted to analyze the immune-related pathways involved,and protein-protein interaction networks were used to assess functional associations.Finally,drug enrichment analysis and molecular docking were performed to predict potential drugs targeting these immune-related genes.RESULTS AND CONCLUSION:Three key immune-related genes-RNASEL,SECTM1,and HSPA6-were identified.These genes were highly expressed in steroid-induced osteonecrosis of the femoral head and exhibited good diagnostic potential,which were involved in multiple immune-related signaling pathways.Mendelian randomization analysis confirmed their potential causal relationship with steroid-induced osteonecrosis of the femoral head.Drug enrichment analysis and molecular docking identified nine potential drugs,including β-ecdysterone,showing the possibility of intervening in the pathological process of steroid-induced osteonecrosis of the femoral head by regulating the HSPA6 protein.These findings provide new biomarkers and drug targets for the early diagnosis and personalized treatment of steroid-induced osteonecrosis of the femoral head.They also highlight the potential application of bioinformatics in Chinese biomedical research,facilitating the integration and translational use of international data in local disease studies.

BACKGROUND:The pathogenesis of steroid-induced osteonecrosis of the femoral head is not yet fully understood.A deep comprehension of the pathology and molecular mechanisms of steroid-induced osteonecrosis of the femoral head,as well as the search for diagnostic markers with high specificity and sensitivity,are crucial for the prevention and treatment of this condition.OBJECTIVE:To identify immune diagnostic markers for steroid-induced osteonecrosis of the femoral head and predict potential drug targets through drug enrichment analysis and molecular docking techniques.METHODS:The study utilized gene expression profile data(GSE123568 and GSE74089)from the GEO databases(a public gene expression database built by the U.S.National Center for Biotechnology Information).R software was used for data normalization and differential gene screening,followed by weighted gene co-expression network analysis(WGCNA)to identify disease-related genes.Immune-related genes were obtained from the GeneCards database and intersected with the differential genes and WGCNA gene sets to select immune-related genes for steroid-induced osteonecrosis of the femoral head.Mendelian randomization was used to validate the potential causal relationship between these immune-related genes and steroid-induced osteonecrosis of the femoral head.Gene Set Enrichment Analysis was conducted to analyze the immune-related pathways involved,and protein-protein interaction networks were used to assess functional associations.Finally,drug enrichment analysis and molecular docking were performed to predict potential drugs targeting these immune-related genes.RESULTS AND CONCLUSION:Three key immune-related genes-RNASEL,SECTM1,and HSPA6-were identified.These genes were highly expressed in steroid-induced osteonecrosis of the femoral head and exhibited good diagnostic potential,which were involved in multiple immune-related signaling pathways.Mendelian randomization analysis confirmed their potential causal relationship with steroid-induced osteonecrosis of the femoral head.Drug enrichment analysis and molecular docking identified nine potential drugs,including β-ecdysterone,showing the possibility of intervening in the pathological process of steroid-induced osteonecrosis of the femoral head by regulating the HSPA6 protein.These findings provide new biomarkers and drug targets for the early diagnosis and personalized treatment of steroid-induced osteonecrosis of the femoral head.They also highlight the potential application of bioinformatics in Chinese biomedical research,facilitating the integration and translational use of international data in local disease studies.

Objective To evaluate the consistency of DNA coding region single nucleotide polymorphism(cSNP)genotyping at the DNA and RNA levels in common body fluid samples based on the next-generation sequencing platform.Methods After extensive literature retrieval,25 cSNP loci of 8 human tissue-specific mRNAs in peripheral blood,semen and vaginal secretion were selected.Two cSNP multiplex genotyping panels based on DNA and RNA,respectively,were developed for use on the MiSeq FGx sequencing platform.45 body fluid samples(including 14 peripheral blood samples,15 semen samples and 16 vaginal secretion samples)were sequenced and analyzed.The inconsistent typing results of DNA and RNA were rechecked by Sanger sequencing.Results The results of cSNP genotyping at the DNA and RNA levels in peripheral blood were completely consistent.Among the 15 semen samples,the genotypes of rs1995640 and rs 1995641 on the TGM4 gene were inconsistent in 3 cases.Among the 16 vaginal secretion samples,there were 2 cases,1 case and 2 case with inconsistent results of rs3869098,rs10947121 and rs12110470 in MUC22 gene,respectively.Conclusion In this study,MiSeq FGx sequencing and Sanger sequencing were used to test 25 cSNP loci with body fluid tissue specificity.The same typing results at the DNA and RNA levels were observed at 20 cSNPs.Inconsistent genotypes at the DNA and RNA levels were observed at 5 cSNPs on the TGM4 and MUC22 genes.This study provides experimental methods and data for forensic cSNP studies.

Objective To explore an assessment model for multi-level integrated training in health service based on advanced intelligent trauma simulators in order to innovate health service training.Methods An assessment model for multi-level integrated training that involved advanced trauma simulators was adopted to assess chains of treatment and rescue that were composed of multi-hierarchy medical institutions.The assessment focused on trauma emergency response capabilities at each level and the overall therapeutic effect.Results In terms of capabilities for treatment and rescue,group B was the best one,followed by group C and group A.As for the overall therapeutic effect,group A was outstanding,followed by group B and group C.Based on the priorities of combat casualty care,the final results of assessment were as follows:group A was the best one,followed by group B and group C.Conclusion The analysis of processes and outcomes of assessment reveals the edge of this model,as evidenced by the continuity of treatment and rescue,integrity of overall effectiveness,and adaptability of this assessment model.

Objective:To construct a full-length infectious clone of enterovirus D68（EV-D68）expressing enhanced green fluorescent protein（EGFP）by reverse genetics in order to provide an efficient tool for studying the biological characteristics and screening antiviral drugs for EV-D68.Methods:Gene synthesis and overlap PCR techniques were used to construct the full-length clone plasmid pUC57-EV-D68 of EV-D68. The full-length viral sequence was then transferred into the pCAGGS plasmid to obtain the pCAGGS-EV-D68 plasmid. The EGFP gene was amplified and inserted into the pCAGGS-EV-D68 plasmid to construct the pCAGGS-EGFP-EV-D68 plasmid. Then，the two constructed plasmids were transfected into human rhabdomyosarcoma（RD）cells to rescue recombinant viruses RV-EV-D68 and RV-EGFP-EV-D68. The rescued viruses were identified using PCR，Western blot，and immunofluorescence techniques. The antiviral effect of doxycycline was evaluated using the rescued RV-EGFP-EV-D68. Statistical analysis was performed using the two independent samples t-test. Results:The recombinant virus RV-EGFP-EV-D68 capable of expressing EGFP was successfully rescued. Even after 15 serial passages，the virus retained EGFP expression with no significant difference in viral titers compared to the parental virus，indicating its stable passage in RD cells. Besides，the rescued strains exhibited similar replication characteristics to the parental virus. While at 24 and 36 h after infection，the titers of the rescued strains were significantly lower than that of the parental strain（both P<0.05）. This study demonstrated that doxycycline significantly reduced the fluorescence intensity of RV-EGFP-EV-D68-infected RD cells（ P<0.01）. Meanwhile，a negative correlation was observed between the doxycycline concentration and the fluorescence intensity，indicating that the rescued virus could be used for antiviral drug evaluation. Conclusions:This study successfully constructs an infectious clone of EV-D68 expressing EGFP. The rescued recombinant virus RV-EGFP-EV-D68 has been verified to be applicable for the evaluation of antiviral drugs.

Objective:To construct a full-length infectious clone of enterovirus D68（EV-D68）expressing enhanced green fluorescent protein（EGFP）by reverse genetics in order to provide an efficient tool for studying the biological characteristics and screening antiviral drugs for EV-D68.Methods:Gene synthesis and overlap PCR techniques were used to construct the full-length clone plasmid pUC57-EV-D68 of EV-D68. The full-length viral sequence was then transferred into the pCAGGS plasmid to obtain the pCAGGS-EV-D68 plasmid. The EGFP gene was amplified and inserted into the pCAGGS-EV-D68 plasmid to construct the pCAGGS-EGFP-EV-D68 plasmid. Then，the two constructed plasmids were transfected into human rhabdomyosarcoma（RD）cells to rescue recombinant viruses RV-EV-D68 and RV-EGFP-EV-D68. The rescued viruses were identified using PCR，Western blot，and immunofluorescence techniques. The antiviral effect of doxycycline was evaluated using the rescued RV-EGFP-EV-D68. Statistical analysis was performed using the two independent samples t-test. Results:The recombinant virus RV-EGFP-EV-D68 capable of expressing EGFP was successfully rescued. Even after 15 serial passages，the virus retained EGFP expression with no significant difference in viral titers compared to the parental virus，indicating its stable passage in RD cells. Besides，the rescued strains exhibited similar replication characteristics to the parental virus. While at 24 and 36 h after infection，the titers of the rescued strains were significantly lower than that of the parental strain（both P<0.05）. This study demonstrated that doxycycline significantly reduced the fluorescence intensity of RV-EGFP-EV-D68-infected RD cells（ P<0.01）. Meanwhile，a negative correlation was observed between the doxycycline concentration and the fluorescence intensity，indicating that the rescued virus could be used for antiviral drug evaluation. Conclusions:This study successfully constructs an infectious clone of EV-D68 expressing EGFP. The rescued recombinant virus RV-EGFP-EV-D68 has been verified to be applicable for the evaluation of antiviral drugs.

Fusarium head blight (FHB), caused by Fusarium graminearum, not only leads to severe yield losses but also poses a threat to food safety due to the mycotoxins produced by the pathogen. Since this disease is preventable but not curable, the current control mainly relies on chemical fungicides, the long-term use of which may lead to pathogen resistance and environmental pollution. To develop green control methods, we screened 13 biocontrol strains from the rhizosphere soil of wheat, among which strain No. 12 (identified as Pythium aphanidermatum) showed significant antifungal effects. In the plate confrontation test, this strain reduced the colony diameter of the pathogen by 69.2% (1.47 mm vs. 4.78 mm in the control group), with an inhibition rate of 77% (P < 0.01). Microscopic observation revealed obvious deformations in the pathogen hyphae, suggesting a lysing effect. The coleoptile experiment further confirmed that the pre-treatment with this strain reduced the incidence rate to 0. These findings provide new candidate strains for the biocontrol of FHB and offer a scientific basis for reducing the use of chemical fungicides and promoting sustainable agricultural development.
Triticum/growth & development* ; Fusarium/growth & development* ; Plant Diseases/prevention & control* ; Soil Microbiology ; Pest Control, Biological/methods* ; Pythium/physiology* ; Biological Control Agents ; Rhizosphere ; Fungicides, Industrial