ObjectiveThis study aims to construct a reconstituted high-density lipoprotein (rHDL) delivery system loaded with kinsenoside (KD@rHDL), and to systematically evaluate its function in enhancing the phagocytosis of amyloid β-protein (Aβ) by microglia and improving the inflammatory state of microglia, as well as to preliminarily explore its potential application value in the treatment of Alzheimer’s disease (AD). MethodsKD@rHDL was prepared by the film hydration method combined with probe sonication and co-incubation. Its morphology was observed by transmission electron microscopy, and the particle size and Zeta potential were measured by dynamic light scattering. The encapsulation efficiency and drug loading were determined by high-performance liquid chromatography. The affinity between KD@rHDL and Aβ was analyzed by surface plasmon resonance (SPR) to assess its feasibility as a medium for Aβ clearance. At the cellular level, after treating mouse microglial cells (BV-2 cells) with KD@rHDL and adding fluorescently labeled Aβ, the phagocytic efficiency of microglia for Aβ was detected by confocal microscopy. Meanwhile, the CCK-8 method was used to evaluate the effect of KD@rHDL on cell viability to determine its safety. The trans-barrier transport ability of KD@rHDL was detected by Transwell assay. The expression levels of NLRP3 inflammasome and downstream inflammatory factor IL-1β in LPS-induced microglia were detected by Western blot to evaluate the regulatory effect of KD@rHDL on the inflammatory state of cells. ResultsCharacterization results showed that the successfully prepared KD@rHDL presented a typical discoid structure under transmission electron microscopy, with a uniform particle size distribution, an average particle size of approximately (14.4±0.24) nm, and a suitable negative Zeta potential, demonstrating good colloidal stability. The drug content determination results indicated that the encapsulation efficiency of KD@rHDL for kinsenoside was (42.24±1.30)%, and the drug loading was (6.03±0.19)%, indicating a good drug loading capacity. The CCK-8 assay results showed that in the set concentration range, the survival rates of BV2 and HT22 cells in the KD@rHDL treatment group were all above 90%, with no significant difference from the control group, indicating good cell safety of the formulation. The results of the Aβ phagocytosis experiment indicated that, compared with the Aβ oligomers (Aβo) group alone, the fluorescence signal intensity within microglia in the KD@rHDL treatment group was significantly enhanced, and a large amount of fluorescence-labeled Aβ was observed to accumulate intracellularly under a fluorescence microscope. The SPR assay results showed that rHDL had a strong affinity for Aβ, with an affinity constant reaching the nanomolar level. Transwell assay results indicated that KD@rHDL could effectively cross the bEnd.3 cell monolayer barrier and be taken up by BV2 and HT22 cells. Western blot assay results showed that high-dose KD@rHDL treatment could significantly reduce the expression level of NLRP3 protein in LPS-induced microglia and simultaneously down-regulate the maturation and secretion of IL-1β, indicating that KD@rHDL can effectively inhibit the activation of the NLRP3 inflammasome pathway and improve the neuroinflammatory state mediated by microglia. ConclusionThis study successfully constructed a reconstituted high-density lipoprotein delivery system loaded with kinsenoside (KD@rHDL). This nano-delivery system not only significantly enhances the phagocytic clearance ability of microglia towards Aβ, but also effectively inhibits the NLRP3/IL-1β-mediated inflammatory pathway, improving the inflammatory state of microglia. The above results indicate that KD@rHDL has a synergistic effect in promoting Aβ clearance and alleviating neuroinflammation, demonstrating potential therapeutic value for AD and providing new ideas and experimental basis for the development of subsequent AD treatment strategies.

Objective:To investigate the mechanism of natural antigen B(nAgB)to protect Immune thrombocytopenia(ITP)mouse model by influencing autophagy.Methods:Twenty-eight female BALB/c mice aged 8-10 weeks were randomly divided into four groups.7 mice of each group were immunized intraperitoneally,the control group was treated with PBS as the control group;ITP group was treated with anti-CD41 monoclonal antibody(anti-CD41Ab)only;nAgB group was treated with nAgB intraperitoneal injection for 5d;nAgB+ITP group was treated with nAgB intraperitoneal injection for 5d,then treated with anti-CD41 Ab.The peripheral blood platelet count in each group was tested;and the spleen and liver should be isolated and weighed,the organ index was calculated;qRT-PCR was used to detect spleen microtubule-associated protein 1 light chain 3(LC3),p62,Beclin-1 mRNA expression levels.Western blot was used to detect the protein expression level of spleen LC3 Ⅱ/LC3 Ⅰ,p62,Beclin-1.Results:Compared with the control group,mice in the ITP group showed a significant decrease in blood PLT count[(102.1±17.9)× 109/L vs(485.4±185.2)×109/L,P＜0.01],a significant increase in spleen index(P＜0.01),mice in the nAgB group showed a significant increase in blood PLT count,rising to(1051±127.6)× 109/L on the 3 day after modeling.Compared with the ITP group,mice in the nAgB+ITP group showed a significant increase in PLT count on the 1 day of anti-CD41 Ab administration[(428.6±131.6)× 109/L vs(102.1±17.9)×109/L,P＜0.05],however,the spleen index was significantly decreased(P＜0.05).qRT-PCR and Western blot results showed that compared with the control group,the mRNA and protein expression levels of spleen LC3,p62 and Beclin-1 were increased in the ITP group of mice(P＜0.05,P＜0.01).Compared with the ITP group,the nAgB+ITP group could significantly decrease mRNA levels of spleen LC3,p62 and Beclin-1(P＜0.05,P＜0.01),and also significantly decrease the protein expression levels of LC3 Ⅱ/LC3 Ⅰ,p62 and Beclin-1(P＜0.05,P＜0.01).Conclusion:nAgB inhibits the transcription and expression levels of autophagy-related genes and regulates immune intolerance,thereby protecting ITP mouse models.

Objective:To analyze the genetic characteristics of the VWF gene c.7332G＞A nonsense mutation and explore its molecular pathogenesis.Methods:Phenotypic diagnosis of the proband was performed using VWF:Ag,VWF:RCo,FⅧ:C and multimeric analysis.The probands were genotyped by NGS whole-exome sequencing,and the sequencing results were validated by sanger sequencing.The family members were genotyped by Sanger sequencing.The VWF gene c.7332G＞A nonsense mutant plasmid was constructed.After transfection,the function of VWF gene c.7332G＞A mutant plasmid was verified at cell level in vitro.The mRNA level was detected by qRT-PCR,and the expression level of protein was detected by Western blot,the function of multimerization was verified by the multimeric analysis.Results:VWF:Ag and VWF:RCo were all less than 3％in the proband,and the multimeric analysis showed multimer deficiency.The proband was diagnosed as type 3 VWD.The homozygous nonsense mutation of VWF gene c.7332G＞A was detected by gene sequencing.The VWF mRNA level of the mutant plasmid was decreased,and the VWF protein expression in the cell supernatant was decreased,the mutant protein was truncated and the function of VWF multimerization was impaired.Conclusion:A homozygous mutation in exon 43 of VWF gene,c.7332G＞A,was responsible for the probands type 3 VWD in the proband.The mutation caused a decrease in the relative level of VWF mRNA and protein,and impaired the function of VWF multimerization.

Objective To screen and analyze key genes in rabbit corneal alkali burns based on transcriptomics se-quencing technology and bioinformatics techniques.Methods Thirty healthy male New Zealand rabbits were randomly di-vided into 2 groups(n=15 per group):The control group received no intervention,while the alkali burn group underwent corneal alkali burn modeling.Histological evaluation of corneal tissues was performed via hematoxylin-eosin(HE)stai-ning.Transcriptome sequencing was conducted for library construction and sequencing.Differentially expressed genes(DEGs)were identified using DESeq2,followed by Gene Ontology(GO)functional enrichment analysis and Kyoto Ency-clopedia of Genes and Genomes(KEGG)pathway analysis.A protein-protein interaction(PPI)network was constructed to screen hub genes,and RT-PCR was employed to validate mRNA expression levels of key genes.Results HE staining revealed orderly arranged corneal stromal layers and scattered stromal cells in the control group,whereas the alkali burn group exhibited stromal edema,thickened collagen fibers with loose/disorganized alignment,and increased fibroblast and inflammatory cell infiltration.Compared to the control group,1 827 significant DEGs were identified in the alkali burn group,including 1 495 upregulated and 332 downregulated genes.GO analysis showed biological processes such as immune response,plasma membrane,and identical protein binding.KEGG analysis indicated that DEGs were enriched in pathways related to cancer,lipid and atherosclerosis,and neuroactive ligand-receptor interaction.The PPI network screened 11 key genes:neutrophil cytosolic factor 1(NCF1),neutrophil cytosolic factor 2(NCF2),matrix metallopeptidase 2(MMP2),ma-trix metallopeptidase 9(MMP9),interleukin-1α(IL-1α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-8(CXCL8),cluster of differentiation 4(CD4),C-C motif ligand 2(CCL2)and tumor necrosis factor(TNF).RT-PCR valida-tion revealed that the mRNA expression levels of key genes in the corneal tissues of the alkali burn group were significantly higher than those in the control group(all P＜0.05),consistent with the transcriptomic sequencing results.Conclusion Based on the rabbit corneal alkali burn model,this study identified 11 key genes(NCF1,NCF2,MMP2,MMP9,IL-1α,IL-1β,IL-6,CXCL8,CD4,CCL2 and TNF)through transcriptomics and bioinformatics analysis.

Fibroblast growth factor,as a critical protein regulating cell growth and differentiation,exhibits aberrant signaling closely associated with various pathological pathologies,including cancer.Among the members of the fibroblast growth factor family,fibroblast growth factor 9(FGF9)has been identified as a critical player in cancer initiation and progression.While numerous studies have investigated the molecular mechanisms of FGF9 individually,comprehensive reviews addressing its impact in cancer remain scarce.This article systematically reviews the functional mechanisms and regulatory networks of FGF9 in cancer,with a focus on its roles in common malignancies such as lung cancer,liver cancer,gastric cancer,colorectal cancer,breast cancer,and ovarian cancer.The aim is to facilitate translational research on FGF9 for targeted cancer diagnosis and therapy.

AIM To compare the chemical constituents in traditional decoction and formula granule decoction of classical famous prescription Wendan Decoction.METHODS The HPLC fingerprints were established,after which the contents of adenosine,synephrine,liquiritin,naringin,hesperidin,6-gingerol and adenosine cyclophosphate were determined,cluster analysis,principal component analysis and multidimensional scaling analysis were adopted in the investigation of component differences,and the equivalent of formula granules was adjusted.RESULTS The similarities of HPLC fingerprints for 10 batches of traditional decoctions were higher than those of HPLC fingerprints for 9 batches of formula granule decoctions(P＜0.01).Adenosine,synephrine,liquiritin,hesperidin and cyclic adenosine monophosphate demonstrated higher contents in traditional decoctions than those in formula granule decoctions(P＜0.05),6-gingerol displayed lower content than that in the latter produced by manufacturers A,C(P＜0.05),which was higher than that in the latter produced by manufacturer B(P＜0.01).Various batches of traditional decoctions and formula granule decoctions could be obviously distinguished,adenosine,synephrine and hesperidin exhibited great influences on the classification of principal component analysis,and the quality of formula granule decoctions produced by manufacturer C was closer to that of traditional decoctions.After equivalent correction,the contents of various constituents in formula granule decoctions produced by manufacturers A,C showed no significant differences as compared with those in traditional decoction(P＞0.05).CONCLUSION The formula granules of Wendan Decoction from different manufacturers exist quality differences,so the preparation process and extraction process of this preparation should be optimized to improve quality,and equivalent ratio should be adjusted according to actual requirements to ensure its scientific and rational clinical application.

Objective To investigate the distribution and antimicrobial resistance profiles of common pathogens isolated from cerebrospinal fluid(CSF)in CHINET program from 2015 to 2021.Methods The bacterial strains isolated from CSF were identified in accordance with clinical microbiology practice standards.Antimicrobial susceptibility test was conducted using Kirby-Bauer method and automated systems per the unified CHINET protocol.Results A total of 14 014 bacterial strains were isolated from CSF samples from 2015 to 2021,including the strains isolated from inpatients(95.3％)and from outpatient and emergency care patients(4.7％).Overall,19.6％of the isolates were from children and 80.4％were from adults.Gram-positive and Gram-negative bacteria accounted for 68.0％and 32.0％,respectively.Coagulase negative Staphylococcus accounted for 73.0％of the total Gram-positive bacterial isolates.The prevalence of MRSA was 38.2％in children and 45.6％in adults.The prevalence of MRCNS was 67.6％in adults and 69.5％in children.A small number of vancomycin-resistant Enterococcus faecium(2.2％)and linezolid-resistant Enterococcus faecalis(3.1％)were isolated from adult patients.The resistance rates of Escherichia coli and Klebsiella pneumoniae to ceftriaxone were 52.2％and 76.4％in children,70.5％and 63.5％in adults.The prevalence of carbapenem-resistant E.coli and K.pneumoniae(CRKP)was 1.3％and 47.7％in children,6.4％and 47.9％in adults.The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)and Pseudomonas aeruginosa(CRPA)was 74.0％and 37.1％in children,81.7％and 39.9％in adults.Conclusions The data derived from antimicrobial resistance surveillance are crucial for clinicians to make evidence-based decisions regarding antibiotic therapy.Attention should be paid to the Gram-negative bacteria,especially CRKP and CRAB in central nervous system(CNS)infections.Ongoing antimicrobial resistance surveillance is helpful for optimizing antibiotic use in CNS infections.

Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3％of the total number of clinical isolates from all patients.Overall,21.8％of the 514 715 strains were gram-positive bacteria,and 78.2％were gram-negative bacteria.Majority(90.9％)of the strains were isolated from inpatients.About 42.9％of the strains were isolated from respiratory specimens,and 22.9％were isolated from urine.More than half(60.7％)of the strains were isolated from male patients,and 39.3％isolated from females.About 51.1％of the strains were isolated from patients aged 65-＜75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8％in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2％,1.5％)and linezolid-resistant strains(3.4％,0.3％)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3％,4.0％,and 1.7％in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2％)to imipenem and meropenem were 20.9％and 22.3％,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7％-7.8％of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6％)to imipenem and meropenem were 68.4％and 70.6％respectively,while 28.5％and 24.3％of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-＜75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.

Objective:To compare the therapeutic efficacy and safety of local anesthesia and spinal anesthesia for the patients with varicocele(VC)who underwent microsurgical varicocelectomy(MV).Methods:We retrospectively analyzed the data of VC patients who underwent MV treatment at the Andrology Department of the Affiliated Ruikang Hospital of Guangxi University of Chinese Medicine from May 2020 to March 2023.Cases with complete clinical data and follow-up evaluation were selected and divided into a control group(spinal anesthesia)and an observation group(local anesthesia)according to different anesthesia methods.The surgical time(including anesthesia time),visual analogue scale(VAS)score for pain,hospital stay,treatment cost,sperm concentration,for-ward motile sperm rate,and normal sperm morphology rate after three months of surgery,as well as postoperative complications and re-currence rate were compared between the two groups.Results:A total of 107 eligible cases were included,with 56 cases in the con-trol group and 51 cases in the observation group.There was no significant difference in the VAS score for pain during and after four hours of surgery,as well as postoperative complications,and recurrence rate between the two groups(P＞0.05).There was an signif-icant increase in sperm concentration,forward motile sperm rate,and normal sperm morphology rate in both of two groups after three months of surgery(P＜0.05).However,there was no significant difference between the two groups three months after surgery(P＞0.05).The surgical time and hospital stay were shorter than those of the control group(P＜0.05).And the treatment cost in observa-tion group was lower than that of the control group(P＜0.05).Conclusion:Both local anesthesia and lumbar anesthesia for MV treatment of VC have good efficacy and safety.However,patients treated with MV under local anesthesia for VC have obvious advanta-ges in terms of operation time(including anesthesia time),hospital stay,and treatment cost,which is worthy of clinical promotion and application.

Objective To investigate the distribution and antimicrobial resistance profiles of common pathogens isolated from cerebrospinal fluid(CSF)in CHINET program from 2015 to 2021.Methods The bacterial strains isolated from CSF were identified in accordance with clinical microbiology practice standards.Antimicrobial susceptibility test was conducted using Kirby-Bauer method and automated systems per the unified CHINET protocol.Results A total of 14 014 bacterial strains were isolated from CSF samples from 2015 to 2021,including the strains isolated from inpatients(95.3％)and from outpatient and emergency care patients(4.7％).Overall,19.6％of the isolates were from children and 80.4％were from adults.Gram-positive and Gram-negative bacteria accounted for 68.0％and 32.0％,respectively.Coagulase negative Staphylococcus accounted for 73.0％of the total Gram-positive bacterial isolates.The prevalence of MRSA was 38.2％in children and 45.6％in adults.The prevalence of MRCNS was 67.6％in adults and 69.5％in children.A small number of vancomycin-resistant Enterococcus faecium(2.2％)and linezolid-resistant Enterococcus faecalis(3.1％)were isolated from adult patients.The resistance rates of Escherichia coli and Klebsiella pneumoniae to ceftriaxone were 52.2％and 76.4％in children,70.5％and 63.5％in adults.The prevalence of carbapenem-resistant E.coli and K.pneumoniae(CRKP)was 1.3％and 47.7％in children,6.4％and 47.9％in adults.The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)and Pseudomonas aeruginosa(CRPA)was 74.0％and 37.1％in children,81.7％and 39.9％in adults.Conclusions The data derived from antimicrobial resistance surveillance are crucial for clinicians to make evidence-based decisions regarding antibiotic therapy.Attention should be paid to the Gram-negative bacteria,especially CRKP and CRAB in central nervous system(CNS)infections.Ongoing antimicrobial resistance surveillance is helpful for optimizing antibiotic use in CNS infections.