The codon was optimized for the bovine nebovirus(BNeV)VP1 gene and the recombi-nant plasmid pFastBac-Dual-VP1 was constructed,and BNeV-VP1 virus-like particles(VLPs)were prepared using a baculovirus expression system,and identified by Western blot,indirect im-munofluorescence and electron microscopy.Successfully validated VLPs were mixed with MF59 adjuvant and CpG-ODG,and mice were immunised by intramuscular injection and evaluated for immunity effects.The results showed that the optimized CAI(codon adaptation index)of VP1 gene was 0.93 and the GC content was 60.4％.The constructed recombinant plasmid was trans-formed into DH10Bac for blue-white spot screening,and after successful verification,it was trans-fected into SF9 cells to obtain recombinant baculovirus Baculo-BNeV-VP1.BNeV virus-like parti-cles with diameters ranging from 35 to 40 nm were observed under the electron microscope,and both IFA and Western blot experiments proved that the target proteins were successfully ex-pressed and biologically active,and protein optimisation revealed that the highest protein expres-sion was found at the infectious dose MOI=0.5.Mice were immunized by intramuscular injection after 50 μg of VLPs were mixed with MF59 adjuvant and CpG-ODN.The results showed that the VLPs immunization group produced IgG antibodies 7 days after the first dose,and the antibody ti-ter increased gradually,reaching a maximum of 1∶102 400,and declined at 35 d,but still main-tained a high level;The blocking titer BT50 is up to 640,which can induce the production of BNeV VP1-specific blocking antibody in mice.In this study,the baculovirus expression system was used to express the VP1 protein of BNeV,and BNeV VLPs were successfully constructed,which could induce humoral immune response in mice,which provided a reference for the follow-up research of BNeV vaccine.

Codon optimization was performed for the M and HN genes of bovine parainfluenza virus type 3(BPIV3),and the recombinant shuttle plasmid Dual-M+HN was constructed.BPIV3 VLPs was prepared using the baculovirus expression system,and verified by Western blot,IFA and elec-tron microscopy.The successfully verified virus-like particle(VLPs)were mixed with MF59 adjuvant and CpG-ODN immunoenhancer to immunize mice by intramuscular-injection,and BPIV3 inactivated vaccine group and adjuvant control group were set up.The immune effect of BPIV3 VLPs was evaluated by monitoring mouse serum specific antibodies,neutralizing antibodies and hemagglutination inhibition antibodies.The results showed that the optimized codon adaptation in-dex(CAI)of the M and HN protein genes were 0.96 and 0.95,respectively,and the CG content reached 54.1％and 53.1％,respectively.The constructed recombinant plasmid was transformed in-to DHI0Bac for blue and white spot screening.The validated recombinant rod particles were trans-fected into Sf9 cells to obtain the rod-shaped virus pFastBac-M+HN.Under electron microscopy,BPIV3 VLPs with a diameter of approximately 180 nm were observed.IFA and Western blot ex-periments confirmed the successful expression and biological activity of the target protein.Through protein optimization,it was found that the protein expression was highest at an infection dose of MOI=5.After mixing 50 μg VLPs with MF59 adjuvant and CpG-ODN,mice were immunized by intramuscular injection.The results showed that the antibodies in the VLPs immunized group be-gan to rise at 2 weeks of the first immunization and reached their peak at 21 days of the second im-munization,with an average IgG antibody titer of 1∶40 228;The average titer of neutralizing anti-bodies is 1∶298;The titer of hemagglutination inhibition antibody is 1∶549,reaching the level of inactivated vaccine(P≥0.05),indicating that the VLPs prepared in this experiment can induce hu-moral immune response in the body.In summary,this study successfully prepared VLPs capable of self-assembly expression of BPIV3 HN and M proteins,and induced humoral immune response in mice,providing research basis for subsequent BPIV3 VLPs vaccine research.

BACKGROUND:The technique of ultrasound processing is widely used for molecular biological analysis.It is of great significance to study the DNA quality of tissue with different storage years under new ultrasonic treatment for further specimen quality control of molecular detection.OBJECTIVE:To explore the effects of different storage durations on DNA quality in specimens with ultrasound processing to investigate the optimal storage time for molecular tests.METHODS:Forty specimens of breast biopsy were collected and paraffin specimens were prepared by ultrasonography.These specimens were divided into four groups based on their storage periods:<1 year,1-3 years,>3-5 years,and>5 years,which contained 10 cases in each group.Paraffin specimens were sliced;each slice was 3 μm thick;10-15 slices were taken,and DNA was extracted.The mass concentration of DNA was examined by Nanophotometer N60 ultra-micro spectrophotometer and Qubit 4.0 fluorometer.The purity of the DNA was analyzed by the ratio of A260/A280.DNA fragment integrity was measured by capillary electrophoresis (Qsep 100) to evaluate the quality of the DNA fragments.RESULTS AND CONCLUSION:The mean values of A260/A280 in the four groups were between 1.8 and 2.0,meeting the requirements of tests,without significant differences.The mean values of DNA mass concentration (Qubit concentration) were 30.39,14.33,2.52,and 1.95 ng/μL,respectively.The mean values of the N/Q were 6.48,14.18,24.56,and 29.86.The mean values of DNA were:5.64,1.76,1.24,and 0.80.The percentage of large DNA fragments averaged 56.08％,17.72％,12.68％,and 7.90％.Moreover,the Ct values of the internal control detected by PCR were 15.32,17.09,18.39,and 21.24.The three other groups exhibited significantly lower DNA concentration,higher N/Q ratios,decreased DNA quality and percentage of large fragments,and increased values of Ct,compared with the group of within 1 year of storage (P<0.05).The experimental results suggested that for novel ultrasound processed biopsy specimens,we should prioritize samples stored within 1 year for molecular testing.Samples stored within 3 years can also meet the requirements of second-generation sequencing and other tests.Samples stored within 5 years can only be attempted to carry out PCR.Samples stored for more than 5 years were not recommended to carry out molecular tests.

Sonodynamic therapy (SDT) can potentially induce immunogenic cell death in tumor cells, leading to the release of ATP, and facilitating the initiation of an immune response. Nevertheless, the enzymes CD39 and CD73 can swiftly convert ATP into immunosuppressive adenosine (ADO), resulting in an immunosuppressive tumor microenvironment (TME). This study introduced a nanomedicine (QD/POM1@NP@M) engineered to reprogram TME by modulating the CD39/CD73/ADO pathway. The nanomedicine encapsulated sonosensitizers silver sulfide quantum dots, and the CD39 inhibitor POM1, while also incorporating homologous tumor cell membranes to enhance targeting capabilities. This integrated approach, on the one hand, stimulates the release of ATP via SDT, thereby initiating the immune response. In addition, it reduced the accumulation of ADO by inhibiting CD39 activity, which ameliorated the immunosuppressive TME. Upon administration, the nanomedicine demonstrated substantial anti-tumor efficacy by facilitating the infiltration of anti-tumor immune cells, while reducing the immunosuppressive cells. This modulation effectively transformed the TME from an immunologically "cold" state to a "hot" state. Furthermore, combined with the checkpoint inhibitor α-PDL1, the nanomedicine augmented systemic anti-tumor immunity and promoted the establishment of long-term immune memory. This study provides an innovative strategy for combining non-invasive SDT and ATP-driven immunotherapy, offering new ideas for future cancer treatment.

[This corrects the article DOI: 10.1016/j.apsb.2018.08.009.].

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

(±)-Talapyrones A-F (1-6), six pairs of dimeric polyketide enantiomers featuring unusual 6/6/6 and 6/6/6/5 ring systems, were isolated from the fungus Talaromyces adpressus. Their structures were determined by spectroscopic analysis and HR-ESI-MS data, and their absolute configurations were elucidated using a modified Mosher's method and electronic circular dichroism (ECD) calculations. (±)-Talapyrones A-F (1-6) possess a 6/6/6 tricyclic skeleton, presumably formed through a Michael addition reaction between one molecule of α-pyrone derivative and one molecule of C₈ poly-β-keto chain. In addition, compounds 2/3 and 4/5 are two pairs of C-18 epimers, respectively. Putative biosynthetic pathways of 1-6 were discussed.
Polyketides/isolation & purification* ; Talaromyces/chemistry* ; Stereoisomerism ; Molecular Structure ; Circular Dichroism ; Pyrones/chemistry*

Nanozymes are a distinct category of nanomaterials that exhibit catalytic properties resembling those of enzymes such as peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Nanozymes derived from Chinese herbal medicines exhibit the catalytic functions of their enzyme mimics, while retaining the specific medicinal properties of the herb (termed "herbzymes"). These nanozymes can be categorized into three main groups based on their method of synthesis: herb carbon dot nanozymes, polyphenol-metal nanozymes, and herb extract nanozymes. The reported catalytic activities of herbzymes include POD, SOD, CAT, and GPx. This review presents an overview of the catalytic activities and potential applications of nanozymes, introduces the novel concept of herbzymes, provides a comprehensive review of their classification and synthesis, and discusses recent advances in their biomedical applications. Furthermore, we also discuss the significance of research into herbzymes, including the primary challenges faced and future development directions.
Nanostructures/chemistry* ; Humans ; Herbal Medicine/methods* ; Superoxide Dismutase/chemistry* ; Catalase/chemistry* ; Drugs, Chinese Herbal/chemistry* ; Catalysis ; Glutathione Peroxidase/chemistry*

Objective To investigate the changes of fibrinogen/albumin ratio(FAR)and uric acid/high-density cholesterol ratio(UHR)in elderly patients with type 2diabetic kidney disease(DKD),as well as the value of the above indexes for early diagnosis of DKD.Methods 310 elderly type 2diabetes mellitus(T2DM)patients were selected and divided into 106 cases in the simple T2DM group,121 cases in the early DKD group and 83 cases in the clinical DKD group according to the urinary microalbumin/urine creatinine ratio(UACR),and the general data and laboratory data of the three groups were compared.Results With the increase of proteinuria,FAR and UHR were significantly elevated,and FAR and UHR were higher in the early DKD group and clinical DKD group than in the sim-ple T2DM group,and FAR was higher in the clinical DKD group than in the early DKD group,and the differences were statistically signif-icant(P＜0.05).Spearman's correlation analysis showed that both UHR and FAR were positively correlated with UACR(r=0.174,P=0.002;r=0.445,P＜0.001),UA(r=0.783,P＜0.001;r=0.147,P=0.010),Cr(r=0.392,P＜0.001;r=0.332,P＜0.001),and negatively correlated with eGFR(r=-0.114,P=0.045;r=-0.374,P＜0.001).In addition,UHR was positively correlated with FAR(r=0.147,P=0.010).The results of multiple Logistic regression analysis showed that long disease course,high glycated hemoglo-bin level,high systolic blood pressure,high FAR,and high UHR were risk factors for the occurrence of DKD(P＜0.05);among which long disease course,high FAR,and high UHR were risk factors for the occurrence of early DKD(P＜0.05).ROC curve analysis sugges-ted that FAR and UHR combined with disease duration had an important diagnostic value for the occurrence of early DKD(AUC=0.803,95％CI:0.752-0.853).Conclusion Elevated FAR and UHR are closely related to the development of DKD and may be independent risk factors for the occurrence of early DKD.Monitoring FAR and UHR in combination with urine microalbumin testing is benefical for the early diagnosis of DKD.

Young management talents are the driving force behind the refined management of hospitals and the future and hope of high-quality development.In the context of the new era,high-quality development has become the main theme of reform and development in public hospitals,which puts forward new and higher requirements for the cultivation of young management tal-ents in hospitals.Based on literature research,this article systematically summarizes and explores the concept and characteristics of young management talents in hospitals.Taking Jiangsu Provincial People's Hospital as an example,it actively supports and serves the growth and development of young management talents through specific practices such as optimizing organizational man-agement,improving training systems,strengthening incentive mechanisms,and smoothing development channels.These practices have achieved good results and aim to provide references for the cultivation and development of young management talents in pub-lic hospitals in China.