ObjectiveThis study aims to systematically analyze the blood-absorbed components and metabolic profiles of Xiebaisan（XBS） in normal and chronic bronchitis （CB） mice using ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）， while comparing differences between the two states. MethodsThirty female BABL/c mice were randomly divided into the normal group， the normal drug administration group， the CB group， the CB drug administration group and the dexamethasone group， with 6 mice in each group. The CB mouse model was established by inducing with ovalbumin （OVA）. The mice in the normal drug administration group and the CB drug administration group started to be gavaged with XBS（13.2 g·kg^-1） from the 21^st day， and the dexamethasone group mice were simultaneously gavaged with dexamethasone （0.5 mg·kg^-1） until the end of the 35^th day of the experiment. Subsequently， serum samples were collected and evaluated for their efficacy， based on the pharmacological evaluation indicators， to determine the efficacy of XBS in treating CB. Then the UPLC-Q-Exactive Orbitrap MS was employed to identify and analyze the chemical constituents， blood-absorbed components， and metabolites of XBS. Chemometric analysis was conducted to reveal metabolic profile differences under "dual states". Concurrently， Real-time PCR technology was utilized to detect the expression levels of key liver metabolic enzymes CYP2E1， CYP3A1， UGT1A1， and UGT1A6. ResultsA total of 28 prototype components and 158 metabolites （including 48 phase Ⅰ metabolites and 110 phase Ⅱ metabolites） of XBS were unambiguously identified in the serum of normal mice. Additionally， a comprehensive characterization was performed on a total of 32 prototype components and 178 metabolites （including 50 phase Ⅰ metabolites and 128 phase Ⅱ metabolites） of XBS in the serum of CB mice. Among them， 27 prototype components were detected in both states， including 12 flavonoids， 2 alkaloids， 3 triterpenes， 4 organic acids， 3 amides， 1 stilbene and 2 other compounds. The chemometrics analysis revealed no significant difference in the prototype components and metabolites of XBS between normal and CB mice； however， there was a significant increase in the in-vivo exposure of XBS in CB mice. Compared to normal mice， the levels of phase Ⅰ metabolites such as oxidation， reduction and methylation of blood components of XBS as well as phase Ⅱ metabolites of glucuronidation showed significant changes in CB mice. Real-time PCR further confirmed that these alterations were attributed to the upregulation of CYP2E1 （P<0.05）， CYP3A1 （P>0.05）， UGT1A1 （P<0.01） and UGT1A6 （P<0.01） enzymes expression in the liver of CB mice. ConclusionThis study elucidated the disparities in the levels of the blood-absorbed components and metabolic profiles of XBS in normal and CB mice， especially in oxidation， reduction， methylation in phase Ⅰ metabolism and glucoaldehyde acidification in phase Ⅱ metabolism. And there are related to the differences in the expression levels of phase Ⅰ and phase Ⅱ metabolic enzymes CYP2E1， CYP3A1， UGT1A1 and UGT1A6 in the liver.

Objective To analyze the single nucleotide polymorphism (SNP) and copy number variation (CNV) of cardiac myxoma to find the SNP sites and CNV events that may play important roles in the occurrence of tumors. Methods The patients with myxoma admitted to our hospital from 2015 to 2019 were randomly selected. The SNP analysis and the CNV test in gene level were performed through whole exome sequencing (WES). The samples were divided into two groups according to the mean size of the tumor: a diameter≤5.7 cm group and a diameter>5.7 cm group. The analysis results were compared between the two groups. Results A total of 14 patients were enrolled, including 8 females and 6 males with a mean age of 61.4 (41-79) years. Thirty-seven cancer-genes with SNP were detected, among which 18 mutated sites had a mutation rate of>10%; and TP53, EP300 and CREBBP played a core binding role in protein-protein interaction-network. The GO enrichment results showed significant differences in the regulation of cell secretion of the mutated genes, and the KEGG enrichment results showed significant differences in the PI3K-AKT and JAK-STAT signaling pathways in the occurrence of myxoma. In addition, 17 new mutation sites of tumor genes with high mutation effect were found in SNP detection. The WES results of 14 samples showed that the CNV events were detected in 120 tumor genes of the samples, 10 of which were included in two tumor databases. The GO enrichment results showed significant differences in the tube development and regulation of cell proliferation, and the KEGG enrichment results showed significant differences in the comprehensive tumor signaling pathway. Statistical differences of ERCC6L and INTS6L in CNV test were found (P=0.030). Conclusion There may be multiple tumor gene site mutations in the process of tumor generation, among which there are multiple core tumor genes such as TP53, EP300 and CREBBP, regulating tumor cells through PI3K-AKT and JAK-STAT signaling pathways and playing an important role in tumor generation. The CNV of ERCC6L and INTS6L genes may be related to tumor growth.

In order to delve into the molecular mechanism underlying the increased pathogenicity of the recombinant Muscovy duck parvovirus(rMDPV)towards Muscovy ducklings,two sub-genom-ic fragments of the rMDPV strain ZW were cloned into the plasmid pBluescript Ⅱ(SK)to gener-ate the recombinant plasmid pZW.A single nucleotide mutation was engineered in the VP3 gene of pZW to discriminate from the parental strain ZW.pZW plasmid-lipid complex was transfected into the chorioallantoic membrane of 11-day-old embryonated Muscovy duck embryos,resulting in res-cue of infectious virus,rZW,carrying the genetic marker.The rescued virus was passaged in 12-day-old embryonated Muscovy duck embryos and exhibited the similar medium embryo lethal dose(ELD50)value and growth curve compared to the parental strain ZW.Both rZW and strain ZW led to 100％mortality in the infection tests performed with 3-day-old Muscovy duckling.Postmortem necropsy revealed a characteristic intestine embolism formed in the rZW-infected ducklings.Taken together,the generation of the infectious clone pZW lays a solid foundation for deciphering the pathogenesis of rMDPV.

Objective:To investigate the application value of dual-layer detector spectral CT in the precise outlining of gross tumor volume (GTV) in lung cancer patients.Methods:Imaging data of 39 patients with pathologically confirmed lung cancer on dual-energy enhanced CT scans in Hebei Medical University Third Hospital were retrospectively analyzed. Among them, 13 patients were not complicated with lung atelectasis and 26 cases were complicated with lung atelectasis and 9 of them received positron emission computed tomography (PET-CT) scan. The virtual single-energy images of arterial and venous dual-phase 40 keV images were reconstructed with the spectral base images of Iqon dual-energy CT, and the GTV of the primary foci was outlined using the reconstructed images and conventional enhanced CT images. The GTV outlined by conventional enhanced CT image, 40 keV virtual monoenergetic (VM) CT image, 40 keV VM-iodine density (VM-ID) fusion image in the arterial phase, conventional enhanced CT image, 40 keV VM image and 40 keV VM-ID image in the venous phase and PET-CT image was defined as GTV ACT, GTV A40VM, GTV A40VMID, GTV VCT, GTV V40VM, GTV V40VMID and GTV PET-CT, respectively. The consistency of target area outlining was assessed by calculating the GTV volume, the Dice similarity coefficient (DSC), and the 95 th percentile of the Hausdorff distance (HD95). Pairwise comparison among groups was conducted by Friedman test and corrected by Bonferroni correction. Results:In GTV comparisons, the differences in GTV ACT, GTV VCT, GTV A40VM and GTV V40VM in patients without pulmonary atelectasis were not statistically significant ( χ2=1.89, P=0.595). The DSC and HD95 of GTV ACTvs. GTV A40VM were 0.96 and 3.00, and the DSC and HD95 of GTV VCTvs. GTV V40VM were 0.94 and 2.93, respectively. The differences in GTV ACT, GTV A40VM, GTV A40VMID, GTV VCT, GTV V40VM, GTV V40VMID and GTV PET-CT in patients complicated with pulmonary atelectasis were statistically significant (all P<0.001). Pairwise comparison of Bonferroni correction showed that there was no statistically significant difference in GTV A40VM, GTV A40VMID, GTV V40VM, GTV V40VMID and GTV PET-CT (all P=1.000), all of which were significantly smaller than those of GTV ACT and GTV VCT (both P=0.001), and there was no statistically significant difference between GTV ACT and GTV VCT (both P=1.000). Based on the tumor extent shown by PET-CT (standardized uptake value =2.5), DSC were slightly higher and HD95 were slightly lower than conventional enhanced CT of GTV A40VM, GTV V40VM, GTV A40VMID, GTV V40VMIDvs. GTV PET-CT, respectively. When the arterial phase sequences were compared with PET-CT, pairwise comparison of Bonferroni correction showed that the DSC and HD95 of GTV A40VMvs. GTV PET-CT and GTV ACTvs. GTV PET-CT were statistically significant (both P<0.01), and the differences were not statistically significant among the remaining groups (all P>0.05). When intravenous phase sequences were compared with PET-CT, pairwise comparison of Bonferroni correction revealed that the DSC and HD95 of GTV V40VMIDvs. GTV PET-CT and GTV VCTvs. GTV PET-CT were statistically significant (both P<0.01), and the differences were not statistically significant among the remaining groups (all P>0.05). Conclusions:The use of 40 keV VMI-ID fusion images to outline the target area of the primary tumor lesions is closer to that of PET-CT, which provides a novel option for the precise outlining of the target area of clinical radiotherapy.

Objective:To investigate how age and nail-tract volumetric bone mineral density (vBMD) are associated with postoperative mechanical performance of proximal femoral nail anti-rotation (PFNA-Ⅱ) fixation for geriatric intertrochanteric fractures using a finite-element analysis.Methods:Fifteen elderly patients with intertrochanteric fracture of the femur were selected for this study. They were 11 females and 4 males and divided into 5 groups based on their ages ( n=3): 55-year-old, 65-year-old, 75-year-old, 85-year-old, and 95-year-old groups. After three-dimensional models of the proximal femur were constructed using the preoperative CT data of their healthy contralateral hip, 31-A1.2 fractures of the AO/OTA type were created and PFNA-Ⅱ fixations simulated. Two loading schemes were created: graded quasi-static axial loads (700 N, 1,400 N, 2,100 N, and 2,800 N) were applied to compute equivalent plastic strain volumes in the femoral head region; displacement-controlled loading was applied to failure to derive load-displacement curves for stiffness and the maximum failure load. Nail-tract vBMD and regional hip vBMDs were measured by quantitative CT. Pearson correlation analysis was conducted to investigate the associations of age and nail-tract vBMD with the aforementioned mechanical indicators. Results:Under the same load, compared with the 55-year-old, 65-year-old, and 75-year-old groups, the plastic strain unit volumes of the fracture models in the 85-year-old and 95-year-old groups increased significantly. Under a load of 700 N, no plastic strain was observed in the fracture models in the 55-year-old, 65-year-old, and 75-year-old groups, while an average plastic strain of approximately 50 mm 3 was observed in the fracture models in the 85-year-old group. Under a load of 2,800 N, the high strain areas in the fracture models in the 85-year-old and 95-year-old groups were mainly concentrated at the tip of the head nail and the junction between the head nail and the main nail. Load-displacement curves showed a marked reduction in the failure load in patients aged ≥85 years. Under loads of 1,400 N, 2,100 N, and 2,800 N, there was a strong association between the nail-tract vBMD and the volume of the plastic strain unit ( r=-0.82, -0.88, -0.89, respectively), which was stronger than those for total-hip vBMD. Conclusions:Finite-element analysis indicates that age and nail-tract vBMD are closely related to local plastic strain and overall stiffness of the proximal femur after PFNA-Ⅱ fixation for the geriatric intertrochanteric fractures. Patients aged ≥85 years old are more prone to plastic yielding, which compromises fixation stability.

Porphyromonas gingivalis is an important pathogenic bacterium causing a variety of oral and systemic diseases. The LuxS/AI-2 quorum sensing system plays a key role in regulating numerous physiological processes such as biofilm formation, virulence factors and drug resistance in Porphyromonas gingivalis. Quorum sensing inhibitors are a new potential antibiotic substitute, a new method to control bacterial infection under the inhibition of biofilm formation, bacterial virulence and no induction of antibiotic resistance. This review is a summary of the research progress of the LuxS/AI-2 quorum sensing system in regulating biofilm formation, virulence, resistance and quorum sensing inhibitors in Porphyromonas gingivalis, and provides a theoretical basis for further research on the inhibitory targets of the LuxS/AI-2 quorum sensing system in Porphyromonas gingivalis.

OBJECTIVE To evaluate the quality differences of Alpinia katsumadai from different habitats. METHODS High- performance liquid chromatography（HPLC） was used to establish the fingerprints of A. katsumadai from 18 batches of different habitats， and the quality of A. katsumadai from different habitats was comprehensively evaluated by similarity evaluation， cluster analysis （CA）， principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA） and the content determination results of alpinetin， pinocembrin， cardamonin and alnustone in A. katsumadai. RESULTS The similarity of HPLC fingerprints for 18 batches of A. katsumadai was ＞0.9. Eleven common peaks were identified from the chromatogram， and four of them were specifically characterized. Both CA and PCA grouped 18 batches of A. katsumadai into 3 categories， extracting 2 principal components （the cumulative variance contribution rate reached 89.798%）. OPLS-DA identified 9 quality difference markers， namely the components corresponding to peaks 4， 9， 3， 2， 7 （pinocembrin）， 8 （cardamonin）， 6 （alpinetin）， 10 and 11 （alnustone）. The content of alpinetin， pinocembrin， cardamonin， and alnustone ranged from 4.507 1-11.579 7， 5.154 4-14.183 3， 5.109 5-13.588 3 and 4.494 6-11.277 2 mg/g， respectively. CONCLUSIONS The quality of A. katsumadai from different habitats is quite different， and the quality of A. katsumadai from Hainan is the best.

Objective To investigate the effect of Biejiajian Pill(BJJP)on malignant biological behavior of hepatocellular carcinoma Hep3B cells and its regulatory mechanism.Methods A total of 72 healthy SD rats were randomly divided into blank control(BC)group,low(0.55 g/kg),medium(1.10 g/kg)and high(2.20 g/kg)BJJP experimental group,and drug-containing serum was prepared.Hep3B cells were divided into BC group,normal rat serum treatment(NC)group,low dose BJJP(LBJJP)group,medium dose BJJP(MBJJP)group and high dose BJJP(HBJJP)group,empty plasmid(pcDNA3.1)group,and CKLF-like MARVEL transmembrane domain containing 6(CMTM6)overexpression(pcDNA3.1-CMTM6)group,and the NC+pcDNA3.1 group,MBJJP+pcDNA3.1 group,NC+pcDNA3.1-CMTM6 group and MBJJP+pcDNA3.1-CMTM6 group.The proliferation of hepatoma Hep3B cells was detected by CCK-8.The migration and invasion of hepatoma Hep3B cells were detected by Transwell assay.The expression levels of proliferation-related proteins proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)related proteins(E-cadherin,N-cadherin and Vimentin),and CMTM6 proteins in hepatoma Hep3B cells were detected by Western blotting experiments.Results Compared with those in BC group,there were no significant differences in the proliferation,migration and invasion abilities of Hep3B cells,or the expression levels of PCNA,EMT related proteins(E-cadherin,N-cadherin and Vimentin)and CMTM6 protein in NC group(P＞0.05).Compared with NC group,LBJJP,MBJJP and HBJJP drug-containing serum inhibited the proliferation,migration and invasion of Hep3B cells,downregulated the protein expression of PCNA;MBJJP and HBJJP upregulated the protein expression of E-cadherin.The protein expressions of N-cadherin,Vimentin and CMTM6 were downregulated,with significant differences(P＜0.05).Compared with pcDNA3.1 group,the protein expression of CMTM6,cell proliferation,migration,invasion,PCNA protein expression,N-cadherin protein expression,and Vimentin protein expression in Hep3B cells in pcDNA3.1-CMTM6 group were significantly upregulated,while the protein expression of E-cadherin was significantly downregulated(P＜0.05).Compared with NC+pcDNA3.1 group,the proliferation,migration and invasion abilities of Hep3B cells in MBJJP+pcDNA3.1 group were decreased,the expression levels of PCNA,Vimentin and N-cadherin protein were decreased,while the expression level of E-cadherin protein was increased.Compared with NC+pcDNA3.1-CMTM6 group,MBJJP+pcDNA3.1-CMTM6 group had the same results in the proliferation,migration and invasion abilities of Hep3B cells and the protein expression levels of PCNA,Vimentin,N-cadherin and E-cadherin.The differences were statistically significant(all P＜0.05).Conclusion BJJP may inhibit the proliferation,migration,invasion and EMT of hepatoma Hep3B cells by regulating the expression of CMTM6.

Colonic mucosal healing is the ultimate goal of ulcerative colitis (UC) treatment, but it remains difficult to realize. Given the higher incidence of UC in males and the beneficial effect of estrogen on UC, we conducted this study to examine the therapeutic potential of estrogen receptor β (ERβ), the primary ER subtype in colon, on mucosal healing in UC. Our study is the first to report that ERβ activation degree was positively correlated with mucosal healing in patients with UC. Furthermore, ERβ activation enhanced mucosal healing in mice with dextran sulfate sodium-induced and biopsy-induced colonic injuries. Mechanistically, ERβ activation promoted autophagy of colonic epithelial cells by inhibiting branched-chain amino acid transport, leading to focal adhesion kinase (FAK) activation. Activated FAK promoted focal adhesion turnover and colonic epithelial cell migration, ultimately facilitating mucosal healing. ERβ ^-/- colitis mice exhibited impaired mucosal healing compared to wild-type littermates, highlighting the crucial effect of ERβ. Importantly, combination with ERβ-agonist diarylpropionitrile enhanced the amelioration of 5-aminosalicylic acid, a standard UC treatment agent, against mouse colitis. These findings attest to the crucial role of ERβ activation in colonic mucosal healing and may further inform the development of novel strategies for UC treatment.

OBJECTIVES: To investigate the expression patterns and prognostic value of lipid metabolism-related genes in breast cancer. METHODS: RNA sequencing data and clinical information were obtained from The Cancer Genome Atlas breast cancer (TCGA-BRCA) dataset, including 1100 breast cancer tissue samples (18 paired with adjacent tissues) and 112 normal breast tissue samples. Differentially expressed lipid metabolism-related genes were screened from a predefined set of 2043 genes using Bioconductor in R, with a false discovery rate <0.05 and |log2(fold change)|>2. Eligible samples were randomly divided into a training cohort (n=651) and a validation cohort (n=431) at a 6∶4 ratio. Prognostic lipid metabolism-related genes were identified using univariate Cox regression (P<0.005) and further refined via LASSO regression. A risk score model was constructed using multivariate Cox regression, and patients were stratified into high- and low-risk groups based on the median risk score. The model's performance was evaluated using Kaplan-Meier survival analysis with the log-rank test and time-dependent receiver operating characteristic (ROC) curves. A nomogram integrating age, TNM stage, clinical grade, and risk score was developed and validated using calibration curves and the concordance index. Immune cell infiltration was quantified using an immune scoring algorithm, and weighted gene co-expression network analysis (WGCNA) was applied to identify key modules associated with immune cell infiltration. Finally, to validate the function of the key gene ALDH2, small interfering RNA targeting ALDH2 was transfected into breast cancer cells, and its effects on invasion and migration were assessed using Transwell invasion and wound healing assays. RESULTS: A total of 185 differentially expressed lipid metabolism-related genes were identified. Univariate Cox and LASSO regression analyses identified three genes-ALDH2, CYP21A2, and IL24-which were incorporated into the multivariate Cox model. The prognostic model based on these genes demonstrated good predictive performance in both cohorts: patients in the high-risk group had significantly shorter overall survival (P<0.01), and the area under the ROC curve for predicting 1-, 3-, and 5-year survival rates was above 0.64. Analysis of the tumor microenvironment revealed an immunosuppressive phenotype in the high-risk group, characterized by reduced infiltration of several anti-tumor immune cells and downregulation of key immune checkpoint molecules such as PDCD1 and CTLA4. WGCNA suggested an association between ALDH2 and immune cell infiltration. Functional experi-ments confirmed that ALDH2 knockdown significantly enhanced the migration and invasion abilities of breast cancer cells. CONCLUSIONS This study established and validated a pro-gnostic model for breast cancer based on lipid metabolism-related genes. It revealed that low ALDH2 expression is closely associated with poor prognosis and immunosuppression, suggesting its potential as a prognostic biomarker and therapeutic target in breast cancer.