Recent studies have identified a missense mutation in the β1-receptor (ADRB1-A187V) that exerts a pronounced impact on human sleep, with a noted decrease in protein abundance in vivo. The administration of β-blockers is frequently associated with sleep disturbances in clinical settings. In this study, we assessed the influence of various β-blockers on sleep within mouse models. Our findings indicated that β-blockers could induce varying degrees of arousal, sleep disruption, and a decrease in REMS (rapid eye movement sleep). We examined the dose-dependent effects of metoprolol and nebivolol on both sleep and cardiac functionality in both wild-type and Adrb1-A187V mutant mice. Our data suggested that, in contrast to cardiac effects, higher doses of metoprolol are required to have noted impact on sleep. No genotype effect was observed with metoprolol in terms of sleep or cardiac function. In contrast, the mutant mice demonstrated increased sensitivity to nebivolol, which exacerbated sleep fragmentation and impeded the onset of REMS. This study is expected to provide some reference for minimizing the occurrence of sleep disorders and reducing the adverse reactions of drugs to the greatest extent.

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Neoadjuvant therapy for hepatocellular carcinoma is the frontier and hot topic in the current field of liver cancer research.The fundamental purpose is to reduce the risk of postoperative recurrence through standardized preoperative treatment methods.From the attempts of Transcatheter Arterial Chemoembolization monotherapy for neoadjuvant therapy for hepatocellular carcinoma to systematic treatment represented by"targeted combined with immunotherapy",the latter has become the most promising neoadjuvant strategy due to its high objective response rate and potential to induce pathological complete remission.However,the field still faces challenges such as lack of evidence of overall survival benefit in Phase Ⅲ randomized controlled trials,treatment-related adverse reactions that may lead to delay in surgery,optimal population screening,and timing of surgery.This article aims to briefly discuss the current research status of the application of neoadjuvant therapy in resectable hepatocellular carcinoma,explore relevant diagnosis and treatment concepts,and further understand neoadjuvant therapy.

pAPN is a zinc-dependent metalloprotease,mediating the fusion between virus and host cell,and playing a role as the receptor of coronavirus.To explore the effect of pAPN on PEDV rep-lication,the full-length pAPN gene was amplified from the porcine small intestinal by PCR,and was cloned into the lentiviral vector via the homologous site digested with BamH Ⅰ and Not Ⅰ to obtain the recombinant lentiviral vector PLVX-pAPN-mCMV-ZsGreen1-puro.The recombinant lentiviral vector and helper plasmids pLP1,pLP2,pLP-VSVG were co-transfected into 293T cells for lentiviral packaging.Vero cells were infected with the packaged lentivirus and the pAPN gene overexpressing cells were screened by puromycin.The stable expression of Vero-pAPN monoclonal cell line was screened by a limited dilution method,and the effect of the cell line on the replication of PEDV was determined by qPCR for N mRNA transcription level,Western blot for N protein level,and TCID50.The results showed that the packaged lentivirus could infect Vero cells,and the monoclonal cell line Vero-pAPN(2C5)could stably expressed pAPN.The Vero-pAPN cell line can promote the replication of PEDV,the N gene mRNA transcription level was significantly different at 12-48 h(P＜0.05),the N protein expression level increased,and the TCID50 was significantly different at 24 and 48 h(P＜0.05).In conclusion,the Vero-pAPN cell line was constructed in this study and it can significantly promote the replication of PEDV,which provides a candidate cell line for PEDV vaccine production and isolation.

Danggui Buxue decoction(DBD)is a classic prescription with immunomodulatory and hematopoietic effects.Previous studies have shown the DBD has potential to be used as an oral im-mune booster.However,its immunomodulatory effects and mechanism of action have not been thoroughly studied,especially the protective mechanism of immunomodulatory regulation in the state of immunosuppressive is still unclear.The aim of this study was to explore the protective mechanism of DBD in the immunosuppressive state using network pharmacology combined with animal experiments verification.The active components,core targets and signaling pathways of DBD in treating immunosuppression were obtained using network pharmacology tools.On this ba-sis,the active components of DBD were identified using HPLC-MS,and in vivo studies were con-ducted at the same time.The key active components of DBD obtained using network pharmacology included quercetin,kaempferol and formononetin.The core targets included TP53,RELA,TNF,AKT1,and IL-6.KEGG pathway enrichment analysis showed that phosphoinositide 3-kinase(PI3K)-protein kinase B(AKT)may play an important role in the treatment of immunosuppres-sive diseases using DBD.Molecular docking confirmed that each core target had good binding activ-ity with its corresponding compounds.Animal experiments showed that after DBD intervention,the mRNA gene and protein expression of RELA,TNF,and IL-6 in the serum was significantly down-regulated.The mRNA expression of PI3K and AKT in the ileum and PI3K protein expression were also downregulated.In conclusion,DBD exerts its role in treating immunosuppressive diseases by regulating the PI3K-AKT signaling pathway.

OBJECTIVE To study the constituents from the dried stems and leaves of Illicium dunnianum Tutcher.METHODS The compounds were isolated and purified by column chromatography of AB-8 macroporous resin,silica gel,HW-40C,ODS,Sephadex LH-20,and semi-preparative RP-HPLC.Their structures were elucidated by physicochemical properties,spectral analyses and ECD.RESULTS The 70%ethanol extract of Illicium dunnianum was subjected to AB-8 macroporous adsorption resin CC to yield 30%ethanol fraction.Five compounds were obtained and characterized as anisole glycol-7-O-β-L-funanarabifuranosyl-(1→6)-β-D-glucopyranoside(1),4-O-β-D-glucopyranosyloxy-benzaldehyde(2),benzyl-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside(3),β-D-glucopyranoside benzoate(4)and sachalinoside B(5),respectively.CONCLUSION Compound 1 is a new phenolic glycoside,2-5 are identified from Illicium dunnianum for the first time.

Objective:To investigate the dosimetric advantages of deep inspiration breath-hold (DIBH) technique in postoperative radiotherapy for left-sided breast cancer.Methods:A prospective study was conducted on patients requiring adjuvant radiotherapy after left-sided breast cancer surgery at Jiangsu Cancer Hospital from January 2018 to May 2023. CT simulation images were acquired under both free breathing (FB) and DIBH respiratory modes. Planning target volumes (PTV) and organs at risk (OAR) were delineated, and dosimetric parameters were compared between the two respiratory modes. Additionally, patients were grouped into subgroups [internal mammary lymph node irradiation (IMNI) vs. non-IMNI, breast-conserving surgery (BCS) followed by radiotherapy vs. modified radical mastectomy (MRM) followed by radiotherapy], and dosimetric differences among subgroups for both breathing modes were compared. The Velocity system was used to measure the minimum distances from the heart and left anterior descending coronary artery (LAD) to the PTV surface on CT images. These distances were defined as the heart-to-PTV and LAD-to-PTV distances. Pearson correlation analysis was performed to assess the relationships between heart D max and LAD D max, heart-to-PTV distance and heart D mean, and LAD-to-PTV distance and LAD D max under both respiratory modes. Results:A total of 132 patients were included. Compared to the FB, DIBH showed no significant difference in target dose distribution, but significantly reduced dose to OAR. Specifically, the heart D mean and D max decreased by 1.8 Gy and 8.1 Gy, respectively, and the LAD D max decreased by 7.9 Gy, and the affected lung V 5 Gy and V 20 Gy were reduced by 6.4% and 2.5%, respectively (all P<0.05). All subgroups benefited from DIBH, with greater decrease of dose to OAR in the IMNI subgroup (compared with the non-IMNI subgroup) and the subgroup of MRM followed by radiotherapy (compared with the BCS followed by radiotherapy group). Under both FB and DIBH modes, heart D max and LAD D max showed linear correlations ( r=0.62 and 0.84, respectively; both P<0.001), heart-to-PTV distance correlated with heart D mean ( r=-0.61 and -0.67, respectively; both P<0.001), and LAD-to-PTV distance correlated with LAD D max ( r=-0.58 and -0.63, respectively; both P<0.001). Conclusions:The DIBH technique can significantly reduce dose to the heart, LAD, and lungs in patients undergoing postoperative radiotherapy for left-sided breast cancer without compromising target dose. Patients receiving IMNI after left-sided breast cancer surgery benefit more from the DIBH technique.

Objective:To explore the prognosis of patients with gastric cancer peritoneal metastasis (PM) receiving programmed cell death-1 (PD-1) antibody therapy, and investigate the biomarkers that affect the prognosis of anti-PD-1 therapy.Methods:This restrospecific study collected the clinic-pathological data of 56 patients with peritoneal metastasis of gastric cancer who received first-line treatment in the Nanjing Drum Town Hospital from March 2020 to September 2023, among which 41 had received anti-PD-1 immunotherapy and 15 hadn't. The relationship between overall survival (OS) and anti-PD-1 immunotherapy was evaluated by Kaplan-Meier analysis. The relationship between baseline peripheral blood indicators and treatment response of patients with anti-PD-1 treatment was analyzed using unpaired t-test. Subsequently, the Cox proportional risk regression model was used to explore the clinical prognostic factors that may affect anti-PD-1 immunotherapy by univariate and multivariate analysis. The clinical prognostic factors included baseline data and baseline peripheral blood indexes such as anti-PD-1 treatment lines, Eastern Cooperative Oncology Group performance status (ECOG PS), combined positive score (CPS), expression of human epidermal growth factor receptor 2 (Her-2), EBER status, pathological types, other metastatic lesions, ascites content before immunotherapy, with or without abdominal drainage during anti-PD-1 treatment, blood lipid indicators, inflammatory indicators, and tumor indicators. Results:Kaplan-Meier survival statistics showed similar OS (15.9 vs. 15.2 months, P=0.600) in patients with anti-PD-1 therapy compared to those without anti-PD-1 therapy. Patients with baseline high-density lipoprotein (HDL) ≥0.97 mmol/L ( n=22) demonstrated a significantly longer median OS compared to those with HDL＜0.97 mmol/L (15.2 vs. 13.5 months; P=0.018). Similarly, the cohort with apolipoprotein A1 (ApoA1) levels ≥0.86 g/L ( n=21) showed superior survival outcomes, with a median OS of 17.7 months versus 12.3 months in the ApoA1＜0.86 g/L group ( n=20; P=0.006). In contrast, elevated baseline alpha-fetoprotein (AFP) levels ( n=2) were associated with markedly reduced survival (median OS: 5.7 vs. 15.2 months in normal AFP group, n=37; P=0.005). Notably, elevated pretreatment ApoA1 levels correlated with enhanced immunotherapy response ( P=0.017). Multivariate Cox regression analysis revealed that ApoA1 deficiency (≥0.86 g/L) independently predicted better OS following PD-1 antibody therapy ( HR=0.35, 95% CI: 0.12-0.98, P=0.046) in gastric cancer patients with PM. Conclusions:In our study, it is first proposed that ApoA1 could be a significant predictor of the survival advantages of immunotherapy in gastric cancer patients with PM.

To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Aim To investigate the regulatory effects of coenzyme Q10(CoQ10)on blood lipid level and intesti-nal flora abundance in atherosclerotic(As)rats.Methods 24 rats were randomly divided into control group,As group and CoQ10 intervention group.Rats in the As group and CoQ10 intervention group were fed with high-fat chow for 2 weeks,combined with abdominal aortic balloon injury to replicate the As model.CoQ10 was administered by gavage start-ing on the next day of modeling,once daily for 4 weeks.Aortic Movat's staining and lipid levels were used to verify the effect of CoQ10 intervention in As,and the abundance of intestinal flora in intestinal contents was analyzed using met-agenomics.Results Compared with the control group,rat aortic tissues in the As group showed endothelial damage,structural disorganization of the internal elastic plate and inflammatory infiltration,serum triglyceride(TG),total cholester-ol(TC)and low density lipoprotein cholesterol(LDLC)levels were increased,and high density lipoprotein cholesterol(HDLC)levels were decreased.Compared with the As group,the structure of the endothelial cells of the aorta and the structure of the endothelial cells,the internal elastic plates and the smooth muscle cell morphology were relatively regular,serum TC,TG and LDLC levels were decreased,and HDLC levels were increased in the CoQ10 intervention group.Com-pared with the control group,the intestinal bacterial biodiversity in the As group was reduced.At the phylum level,the abundance of the Firmicutes and the Bacteroidetes were down-regulated,whereas that of Proteobacteria was up-regulated.At the genus level,the relative abundance of Lactobacillus,Bacteroides,Akkermansia,Limosilactobacillus,Parabacteroides and Ligilactobacillus was down-regulated,and the relative abundance of Muribaculum was up-regulated.Compared with the As group,CoQ10 intervention restored the biodiversity of the intestinal microbiota in As rats and increased the relative abundance of Firmicutes,Bacteroidetes,Lactobacillus,Bacteroides,Akkermansia,Limosilactobacillus,Parabacteroides and Ligilactobacillus,while reducing the relative abundance of Proteobacteria and Muribaculum(all P＜0.05).Conclusion CoQ10 can regulate blood lipid levels in As rats,upregulate the abundance of beneficial microbiota,downregulate the abun-dance of harmful microbiota,and modulate the diversity of the gut microbiota.