Objective The study aimedto investigate the effects and metabolic mechanisms of Gegen Qinlian Decoction(GQD)containing serum on hypoxia-induced glucose metabolism in L02 cells.Methods The effects of five hypoxia durations(6,12,18,24,and 48 hours)on glucose consumption and cell viability of L02 cells were examined under hypoxic conditions to determine the optimal hypoxia time.Normal hepatocytes served as the normal control group.L02 cells with hypoxia-induced reduction in glucose consumption were divided into several groups:hypoxia group,metformin 2 mmol/L group,and 25 g/kg GQD groups treated with 5%,10%,and 15%GQD contain-ing serum.Glucose consumption was used as an indicator of drug efficacy.High-resolution liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was employed to collect metabolite signals from each group.Data were analyzed by using Progenesis QI software,and potential biomarkers were identified through online databases such as HMDB.Finally,metabolic pathways of potential biomarkers were analyzed via the Metabo Analyst 5.0 website.Results An 18 hour hypoxia period was identified as the optimal duration for the replication of the hypoxia-induced L02 cell model.GQD containing serums at 5%and 10%significantly increased glucose consumption in hypoxia-induced L02 cells after 18 hours.14 biomarkers of hypoxia-induced L02 cells were identified,with the levels of 13 biomarkers significantly increased and 1 biomarker significantly decreased.GQD containing serum notably regulated the levels of 3 biomarkers.Conclusion GQD containing serum might improve hypoxia-induced abnormal glucose metabolism in L02 cells and enhance glucose consumption by modulating glycero-phospholipid metabolism,glycosylphosphatidylinositol biosynthesis,and sphingolipid metabolism.

Objective To investigate the regulatory effect of lipocalin 2(LCN2)on epidermal growth factor receptor(EGFR)phosphorylation and its impact on inflammatory damage in human fallopian tube epithelial cells.Methods Human fallopian tube epithelial cells were isolated and a lipopolysaccharide(LPS)intervention was applied to establish an in vitro cell model.The cells were randomly assigned into the following groups:a blank control group(Control),a model group(Model),experimental groups(Model+si-LCN2 or Model+oe-LCN2),and negative control groups(Model+si-NC or Model+oe-NC).Changes in cell viability,apoptosis rates,inflam-matory levels,as well as the expression of EGFR mRNA,LCN2,EGFR,p-EGFR,and the ratio of p-EGFR/EGFR proteins were evaluated.Results Compared to the Model group,the Model+si-LCN2 group exhibited enhanced cell viability,a reduced apoptosis rate,and decreased expression of inflammatory factors(P＜0.05).Immunopre-cipitation analysis confirmed a direct interaction between LCN2 and EGFR.In comparison with the Model group,the Model+oe-LCN2 group demonstrated elevated levels of p-EGFR and the p-EGFR/EGFR ratio(P＜0.05),while no significant change was observed in total EGFR expression(P＞0.05).Conclusion Inhibition of LCN2-mediated EGFR phosphorylation enhances cell viability,reduces apoptosis,and mitigates inflammatory responses,thereby ameliorating LPS-induced inflammatory injury in human fallopian tube epithelial cells.

Objective:To explore the diagnostic key points, treatment strategies, and prognosis of graft-versus-host disease (GVHD) after liver transplantation.Methods:The clinical data of 5 recipients diagnosed with GVHD after liver transplantation at the Liver Transplantation Center of the First Affiliated Hospital of Army Medical University from May 1999 to October 2024 were retrospectively analyzed. The causes, onset, diagnosis, treatment, and prognosis of GVHD after liver transplantation were summarized and analyzed. Literature was searched in CNKI, Wanfang, VIP, Chinese Medical Journal Full-text Database, PubMed, Web of Science, and Google Scholar using the Chinese keywords "移植物抗宿主病+肝移植", and the English keywords "graft versus host disease + liver transplantation". The search time ranged from January 1988 to January 2025. Inclusion criteria for the literature: (1) meeting the clinical or pathological diagnostic criteria of GVHD after liver transplantation; (2) recipient age >18 years; (3) case number ≥2. Exclusion criteria: incomplete clinical data such as incidence, mortality, and clinical manifestations of GVHD after liver transplantation. The retrieved literature was reviewed.Results:All 5 recipients were male. Among them, 4 cases underwent liver transplantation at this center. The incidence of GVHD after liver transplantation in this center was 0.46% (4/872). All 5 cases developed symptoms such as fever, rash, diarrhea, oral ulcers, and pancytopenia on the 19th (5-21) day after liver transplantation. One case had gastrointestinal bleeding. Two cases were diagnosed by skin pathological biopsy, and three cases were diagnosed based on clinical manifestations such as fever, rash, diarrhea, and bone marrow suppression. One case discontinued immunosuppressants, and four cases reduced the dosage of immunosuppressants. Four cases were treated with high-dose glucocorticoids, four with intravenous immunoglobulin (IVIG), three with ruxolitinib, and three with hematopoietic factors. All five cases received protective isolation, anti-infection, and symptomatic supportive treatment. Among the three recipients treated with ruxolitinib, body temperature returned to normal, rash gradually faded, oral ulcers gradually healed, blood cells returned to normal, and they were eventually discharged after recovery. The remaining two cases showed no symptom improvement and died of severe lung infection and multiple organ failure. Literature review A total of 34 articles were included. The incidence of GVHD after liver transplantation was 1.03% (279/27 018), and the onset time ranged from 7 to 1,865 days post-transplantation; 272 cases (97.49%) occurred within 1-8 weeks. The main clinical manifestations included fever (195 cases, 69.89%), rash (267 cases, 95.70%), diarrhea (173 cases, 62.01%), and bone marrow suppression (214 cases, 76.70%). Treatment mainly involved adjustment of immunosuppressants (201 cases, 72.04%), high-dose corticosteroids (215 cases, 77.06%), and IVIG pulse therapy (146 cases, 52.33%). In the end, 83 cases (29.75%) recovered and were discharged, while the mortality rate was 70.25% (196/279), with causes of death including infection, gastrointestinal bleeding, and multiple organ failure.Conclusions:GVHD after liver transplantation has a low incidence, high mortality, and poor prognosis. Diagnosis mainly relies on typical clinical manifestations and pathological results of tissue biopsy. Early administration of high-dose corticosteroids combined with IVIG pulse therapy, timely reduction or discontinuation of immunosuppressants, use of ruxolitinib, active infection management, and enhanced symptomatic and supportive care are effective strategies for treating GVHD after liver transplantation.

Objective·To explore the roles of hyaluronic acid methacryloyl(HAMA)hydrogel in skin wound healing and to characterize the microenvironmental landscape at wound sites.Methods·A full-thickness skin excision model was established in mice,which were randomly divided into a control group(n=3)and a HAMA group(n=3).The wound in the HAMA and control groups were covered with 100 μL of HAMA hydrogel and 100 μL of phenyl-2,4,6-trimethylbenzoylphosphonic acid lithium(LAP),respectively.Both groups were then irradiated with a UV lamp for 20 s.The residual wound areas was measured on days 0,3,7,10,and 14.Wound healing effects of HAMA hydrogel were analyzed by measuring the residual wound area and through H-E staining.Single-cell RNA sequencing technology was utilized to analyze the cellular profile of local wound skin tissues at day 14 post-injury.Immunofluorescence assay was used to detect the levels of type I collagen,type Ⅲ collagen,F4/80,CD206,and CD86 in the wound sites.The mRNA expression levels of Arg1,Nos2,Itgam,and Itgb2 in the mouse macrophage cell line Raw264.7 co-cultured with HAMA hydrogel for 24 h were detected by RT-qPCR.The fibroblasts and macrophages in the local skin of the mouse wound on day 14 were analyzed using the Seurat package,and the communication between fibroblasts and macrophages was analyzed using the CellChat package.Results·Mice treated with HAMA hydrogel exhibited a significantly faster rate of wound healing process compared to the control group.At day 14,wounds in the HAMA-treated mice had already healed,while those in the control group remained unhealed.Single-cell RNA sequencing analysis revealed a remarkable increase in the proportion of fibroblasts in the skin tissues of HAMA-treated wounds.The proportion of the Col3a1-high-expressing fibroblast subset increased(90.2%)compared to the control group(79.8%),while the proportion of the Col1a1-high-expressing fibroblast subset decreased(5.7%vs 15.9%).Immunofluorescence analysis confirmed that the level of type Ⅲ collagen in the wound tissues of the HAMA group was significantly higher than that in the control group(P=0.035),while the level of type Ⅰ collagen was significantly lower(P=0.044).Although there was no significant difference in the proportions of macrophages in the wound tissues between the HAMA-treated and control groups,scRNA sequencing data and in vitro experiments using Raw264.7 cells showed that HAMA hydrogel could induce the expression of Arg1 and decrease the expression of Nos2 in the macrophages(P<0.001).Additionally,macrophages in the HAMA-treated wounds expressed higher levels of CD206 and lower levels of CD86(P=0.042,P=0.011).The results of the CellChat analysis showed that,compared to the control group,increased communication intensity was observed between macrophages and fibroblasts subsets at the wound sites in the mice of HAMA group.Conclusion·The microenvironment after HAMA hydrogel treatment is conducive to skin wound healing,characterized by a local aggregation of anti-inflammatory macrophages and fibroblasts that secrete type Ⅲ collagen.

Objective To investigate the regulatory effect of lipocalin 2(LCN2)on epidermal growth factor receptor(EGFR)phosphorylation and its impact on inflammatory damage in human fallopian tube epithelial cells.Methods Human fallopian tube epithelial cells were isolated and a lipopolysaccharide(LPS)intervention was applied to establish an in vitro cell model.The cells were randomly assigned into the following groups:a blank control group(Control),a model group(Model),experimental groups(Model+si-LCN2 or Model+oe-LCN2),and negative control groups(Model+si-NC or Model+oe-NC).Changes in cell viability,apoptosis rates,inflam-matory levels,as well as the expression of EGFR mRNA,LCN2,EGFR,p-EGFR,and the ratio of p-EGFR/EGFR proteins were evaluated.Results Compared to the Model group,the Model+si-LCN2 group exhibited enhanced cell viability,a reduced apoptosis rate,and decreased expression of inflammatory factors(P＜0.05).Immunopre-cipitation analysis confirmed a direct interaction between LCN2 and EGFR.In comparison with the Model group,the Model+oe-LCN2 group demonstrated elevated levels of p-EGFR and the p-EGFR/EGFR ratio(P＜0.05),while no significant change was observed in total EGFR expression(P＞0.05).Conclusion Inhibition of LCN2-mediated EGFR phosphorylation enhances cell viability,reduces apoptosis,and mitigates inflammatory responses,thereby ameliorating LPS-induced inflammatory injury in human fallopian tube epithelial cells.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

Objective·To explore the roles of hyaluronic acid methacryloyl(HAMA)hydrogel in skin wound healing and to characterize the microenvironmental landscape at wound sites.Methods·A full-thickness skin excision model was established in mice,which were randomly divided into a control group(n=3)and a HAMA group(n=3).The wound in the HAMA and control groups were covered with 100 μL of HAMA hydrogel and 100 μL of phenyl-2,4,6-trimethylbenzoylphosphonic acid lithium(LAP),respectively.Both groups were then irradiated with a UV lamp for 20 s.The residual wound areas was measured on days 0,3,7,10,and 14.Wound healing effects of HAMA hydrogel were analyzed by measuring the residual wound area and through H-E staining.Single-cell RNA sequencing technology was utilized to analyze the cellular profile of local wound skin tissues at day 14 post-injury.Immunofluorescence assay was used to detect the levels of type I collagen,type Ⅲ collagen,F4/80,CD206,and CD86 in the wound sites.The mRNA expression levels of Arg1,Nos2,Itgam,and Itgb2 in the mouse macrophage cell line Raw264.7 co-cultured with HAMA hydrogel for 24 h were detected by RT-qPCR.The fibroblasts and macrophages in the local skin of the mouse wound on day 14 were analyzed using the Seurat package,and the communication between fibroblasts and macrophages was analyzed using the CellChat package.Results·Mice treated with HAMA hydrogel exhibited a significantly faster rate of wound healing process compared to the control group.At day 14,wounds in the HAMA-treated mice had already healed,while those in the control group remained unhealed.Single-cell RNA sequencing analysis revealed a remarkable increase in the proportion of fibroblasts in the skin tissues of HAMA-treated wounds.The proportion of the Col3a1-high-expressing fibroblast subset increased(90.2%)compared to the control group(79.8%),while the proportion of the Col1a1-high-expressing fibroblast subset decreased(5.7%vs 15.9%).Immunofluorescence analysis confirmed that the level of type Ⅲ collagen in the wound tissues of the HAMA group was significantly higher than that in the control group(P=0.035),while the level of type Ⅰ collagen was significantly lower(P=0.044).Although there was no significant difference in the proportions of macrophages in the wound tissues between the HAMA-treated and control groups,scRNA sequencing data and in vitro experiments using Raw264.7 cells showed that HAMA hydrogel could induce the expression of Arg1 and decrease the expression of Nos2 in the macrophages(P<0.001).Additionally,macrophages in the HAMA-treated wounds expressed higher levels of CD206 and lower levels of CD86(P=0.042,P=0.011).The results of the CellChat analysis showed that,compared to the control group,increased communication intensity was observed between macrophages and fibroblasts subsets at the wound sites in the mice of HAMA group.Conclusion·The microenvironment after HAMA hydrogel treatment is conducive to skin wound healing,characterized by a local aggregation of anti-inflammatory macrophages and fibroblasts that secrete type Ⅲ collagen.

Objective:To translate the English version of the Attitude Tool of Delirium (ATOD) into Chinese and test the reliability and validity of its application in Chinese nurses, and to provide an assessment tool for assessing nurses ′ attitudes towards delirium patients. Methods:After obtaining authorization from the source scale developer, the English version of the ATOD scale was translated, back-translated, expert-consulte, pre-surveyed and cross-culturally adapted according to the Brislin translation model. A questionnaire survey was conducted among 298 nurses with experience in caring for patients with delirium in three tertiary hospitals in Jiangsu Province using a convenience sampling method. The critical ratio value and correlation coefficient were used to analyze the scale items. Exploratory factor analysis was used to evaluate the structural validity of the scale, and experts were invited to evaluate the content validity of the scale. Cronbach α coefficient and test-retest reliability were used to evaluate the reliability of the scale. Results:The Chinese version of the ATOD scale contains 3 dimensions of cognitive attitudes, behavioral attitudes, and emotional attitudes toward patients with delirium, with a total of 23 entries. The Cronbach α coefficient for the entire scale was 0.896, with each dimension′s Cronbach α coefficient ranging from 0.614 to 0.912. The retest reliability of the scale was 0.868, with each dimension ′s retest reliability ranging from 0.603 to 0.927. Exploratory factor analysis extracted three eigenvalues with common factors greater than 1, and the cumulative variance contribution rate was 63.812%. Conclusions:The Chinese version of the ATOD scale has good reliability and validity, and can be used as an evaluation tool for evaluating nurses ′ attitudes towards delirium patients in China, so as to provide an effective basis for the formulation of corresponding interventions.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

Periodontitis is a chronic inflammatory disease that affects the tooth-supporting tissues, and it constitutes a major global public health concern. Methylation modifications, including DNA methylation, histone methylation, and RNA m6A modification, represent reversible processes coordinately regulated by methyltransferases, demethylases, and binding proteins. In periodontitis, aberrant methylation modifications suppress Toll-like receptor 2 expression, leading to oral microbial dysbiosis. These modifications further disrupt normal immune regulatory functions through C-C motif chemokine ligands, Fc-γ receptor-mediated phagocytosis, and NF-κB signaling pathways, resulting in localized immune-inflammatory imbalance in periodontal tissues. In addition, various methylation modifications regulate the expression of Runt-related transcription factor 2 (RUNX2), osteoblast-specific transcription factor Osterix (OSX), and receptor activator of nuclear factor-κB ligand (RANKL), thereby interfering with osteoclast and osteoblast differentiation, disrupting bone homeostasis, and ultimately driving alveolar bone resorption. Methylation-related biomarkers demonstrate promising potential for periodontitis screening and prognostic evaluation. While numerous abnormally methylated sites have been identified in periodontitis, the precise signaling pathways and comprehensive epigenetic regulatory networks remain to be fully elucidated. This review systematically summarizes the functional roles of DNA methylation modifications in the pathogenesis of periodontitis and explores their potential value in etiological studies, diagnostic biomarker discovery, and targeted therapeutic interventions, with the aim of providing novel perspectives for periodontitis prevention and treatment strategies.