Objective To investigate the effect of ubiquitin specific peptidase 22 (USP22) on the occurrence and development of esophageal squamous cell carcinoma (ESCC) under hypoxic conditions, and its regulatory relationship with hypoxia-inducible factor-1α (HIF-1α). Methods Western blotting and quantitative polymerase chain reaction were used to detect the differences in USP22 protein and mRNA expression between normal esophageal epithelial cells HEEC and ESCC cell lines KYSE30, KYSE150, EC9706, and TE-1 under normoxic (5% CO2, 20% O2, 75% N2) and hypoxic (5% CO2, 1% O2, 94% N2) conditions. By transfecting USP22 plasmid or siUSP22, ESCC cells were divided into a normoxia control group, a normoxia+USP22 group, a normoxia+siUSP22 group, a hypoxia control group, a hypoxia+USP22 group, and a hypoxia+siUSP22 group. The proliferation and migration abilities of cells in each group were detected. The expression of USP22 and HIF-1α under hypoxic conditions after up-regulating or down-regulating USP22 was detected, and their regulatory relationship was verified. The interaction between USP22 and HIF-1α was verified by co-immunoprecipitation (Co-IP) technique. Results Compared with HEEC cells, the expression of USP22 in ESCC cells was significantly increased (P<0.05). Up-regulation of USP22 expression promoted the proliferation and migration of ESCC cells, while silencing USP22 inhibited the proliferation and migration of ESCC cells (P<0.05). Under hypoxic conditions, the expression of USP22 and HIF-1α increased, and with the up-regulation of USP22 expression, the expression of HIF-1α also significantly increased (P<0.05). Co-IP experiment confirmed the binding between USP22 and HIF-1α. Conclusion Up-regulation of USP22 expression promotes the proliferation and migration of ESCC cells. Hypoxia microenvironment can induce the increase of USP22 expression in ESCC. USP22 may participate in the regulation of the occurrence and development of ESCC by directly binding to HIF-1α.

Background/Aims: Natural killer (NK) cell function is generally considered dampened in chronic hepatitis B virus (HBV) infection; however, the NK cell pool exhibits phenotypic and functional heterogeneity, and the antibody--mediated effect of NK cells remains less characterized. This study evaluated the dynamic changes in antibody-mediated NK cell responses and the involvement of distinct NK subsets across disease stages and during antiviral treatment. Methods: A T-cell receptor-like antibody specific for the HBV core 18–27 peptide (cTCRL-Ab) was used to determine the antibody-mediated effect of NK cells, and an array of NK cell surface markers were analyzed in cross-sectional and longitudinal cohorts of patients with chronic HBV infection. Single-cell RNA sequencing (scRNA-seq) was performed to identify the heterogeneity of NK subsets. Results: The cTCRL-Ab enabled the detection of NK cell cytolytic activity and IFNγ production. Notably, cTCRL-Ab-mediated NK cell responses were compromised in chronically HBV-infected patients, particularly in those receiving pegylated interferon-α (Peg-IFNα), which was associated with the downregulation of CD16 expression. Correspondingly, Peg-IFNα inhibited cTCRL-Ab-mediated NK cell function by reducing CD16 expression in vitro. scRNA-seq revealed that CD16 downregulation occurred mainly within a dysfunctional CD16hi NK subset exhibiting exhaustion properties. In contrast, an activated CD16hiNK subpopulation (CX3CR1⁺KLRC2–CD16hi) with high cytotoxicity was enriched in patients who experienced favorable treatment responses. Furthermore, the intrahepatic CX3CR1+KLRC2–CD16hi subset tended to exhibit functional restoration in HBsAg-loss individuals. Conclusions Our data contribute to the understanding of antibody-mediated responses of NK cells in chronic HBV infection, and highlight a previously unappreciated functional CX3CR1+KLRC2–CD16hiNK subset as a potential therapeutic target.

Based on ancient and modern literature， this paper systematically elucidates the concept， physiological characteristics， and reproductive significance of tiangui （天癸）， and further reviews its theoretical connotation throughout the entire process of male reproductive function， covering its onset， maturation， maintenance， disorder， and decline. In terms of its physiological foundation， tiangui is rooted in kidney essence and regulates male reproductive function through the coordination of zang-fu （脏腑） organs. It possesses characteristics of being time-limited， rhythmic， relatively independent， targeted， and functionally diverse. The occurrence of male reproductive diseases is closely related to the abnormality of tiangui， which primarily manifests as tiangui deficiency and dysregulation of tiangui timing. In clinical practice， treatment should follow the principle of "supplementing its deficiency and regulating its timing"， so as to restore the operating rhythm of tiangui.

ObjectiveTo investigate the feasibility and efficacy of modified video-endoscopic inguinal lymphadenectomy （M-VEIL）， guided by the layer surgery concept， in reducing postoperative complications in patients with penile cancer. MethodsA retrospective analysis was conducted on 27 patients with penile cancer who underwent bilateral laparoscopic inguinal lymphadenectomy at the Affiliated Hospital of Qingdao University between March 2018 and March 2022. All patients had undergone prior penile surgery. M-VEIL followed a standardized four-step procedure guided by fascial planes to achieve compartment-oriented resection， with deep lymphatic trunks ligated by silk sutures and endoscopic compression suture of the skin to the muscle layer using 4-0 absorbable threads. ResultsPreoperative physical examinations and imaging findings revealed bilateral inguinal lymphadenopathy， and no distant metastases were detected in any patient. Among the 27 patients， 12 underwent traditional video-endoscopic inguinal lymphadenectomy （T-VEIL； mean age 61.08±12.75） years and 15 underwent M-VEIL （mean age 54.47±14.64） years. All 27 operations were successfully completed without conversion to open surgery. No significant differences were observed between the two groups in operative time， intraoperative blood loss， postoperative hospital stay， number of lymph nodes retrieved， or lymph node positivity rate （all P >0.05）. Drain removal time was significantly shorter in the M-VEIL group （15.93±6.89 days vs. 23.33±10.42 days； P＜0.05）. Lymphatic leakage （2 vs. 8 patients） and surgical site infection （1 vs. 6 patients） occurred less frequently in the M-VEIL group （both P ＜0.05）. No significant differences were found in lymphocele， flap necrosis， lymphedema， or lower limb edema between the two groups （all P >0.05）. During postoperative follow-up for 1-3 years， 2 patients developed pelvic lymph node metastases， while the remaining 25 showed no evidence of local or pelvic recurrence. ConclusionCompared with T-VEIL， M-VEIL appears to reduce postoperative complications and shorten drain removal time in patients undergoing inguinal lymphadenectomy for penile cancer， while providing favorable clinical outcomes. Preliminary evidence suggests that M-VEIL is a safe and feasible modified procedure.

Objective To investigate the effects of low background radiation environments in deep underground settings on the biological behavior of NP69 human nasopharyngeal epithelial cells(NP69 cells)and the underlying molecular mechanisms.Methods A parallel control experimental design was adopted and NP69 cells were synchronously cultured in settings of three underground depths at the China in situ Deep-Underground Facility&Life Observatory(DeUFO)—ground level(DeUFO-0 m),1 000 m underground(DeUFO-1 000 m),and 1 500 m underground(DeUFO-1 500 m).Changes in cell proliferation and migration capabilities were assessed using the Cell Counting Kit-8(CCK-8)assay and scratch assay,respectively.High-throughput RNA sequencing(RNA-Seq)was performed to identify differentially expressed genes(DEGs).Functional annotation and pathway enrichment analysis of the DEGs were performed using the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.Results CCK-8 assay revealed that,after 72 h of culture,the absorbance value of the DeUFO-0 m group was 1.35 times and 1.27 times those of the those of the DeUFO-1 000 m and DeUFO-1 500 m groups,respectively(both P<0.000 1).After 96 h of culture,the absorbance value of the DeUFO-0 m group was 1.52 times and 1.41 times those of the DeUFO-1 000 m and DeUFO-1 500 m groups,respectively(both P<0.000 1).Colony formation assays revealed that the number of cell colonies in the DeUFO-0 m group was 1.59 times and 1.27 times those in the DeUFO-1 000 m group and DeUFO-1 500 m group,respectively(both P<0.001).The scratch assay revealed that the 36-hour wound healing rate of the DeUFO-0 m group was 2.22 times and 4.00 times those of the DeUFO-1 000 m group and DeUFO-1 500 m group,respectively(both P<0.000 1).Transwell assays revealed that the number of migrating cells in the DeUFO-0 m group was 2.08 times and 2.56 times those in the DeUFO-1 000 m group and DeUFO-1 500 m group,respectively(both P<0.000 1).Transcriptome sequencing analysis revealed consistent upregulation of CELF2,CELF4,CGB8,GRHL2,and DMRTA2 genes in the DeUFO-1 000 m and DeUFO-1 500 m groups.Pathway enrichment analysis indicated significant enrichment of extracellular matrix(ECM)remodeling-associated pathways and gene expression regulation pathways in the experimental groups(false discovery rate[FDR]<0.05).Conclusion The low background radiation environment in deep underground settings suppresses the proliferation and migration activities of NP69 cells by mediating ECM remodeling and post-transcriptional regulatory mechanisms through the regulation of target genes such as the CELF family.This study provides experimental evidence for establishing a dose-response relationship between environmental radiation and cellular effects.

Objective To investigate the impact of various health education intervention strategies on the proper use of personal respiratory protective equipment (RPE) among workers exposed to dust. Methods Dust-exposed workers were recruited from 60 selected enterprises in Guangdong Province using cluster random sampling method. They were randomly allocated to the control, low-intensity intervention, and high-intensity intervention groups, with 358, 346, and 371 workers in each group, respectively. Workers in the control group received no designed intervention. Workers in the low-intensity intervention group received traditional plus mobile health education on the proper use of RPE. Workers in the high-intensity intervention group received all components of the low-intensity intervention, supplemented with peer education. The intervention lasted for six months. RPE usage was compared among the three groups of workers before and after the intervention. Results Workers in the control, low-intensity intervention, and high-intensity intervention groups showed higher rates of both RPE wearing and correct RPE wearing after the intervention than before it within their respective groups (RPE wearing rate: 94.1% vs 99.2%, 95.7% vs 100.0%, 94.6% vs 100.0%, all P<0.01; correct RPE wearing rate: 66.8% vs 91.1%, 67.3% vs 95.7%, 66.6% vs 96.5%, all P<0.01). Post-intervention correct RPE wearing rates were highest in the high-intensity intervention group, followed by the low-intensity intervention group, and the control group, with the percentage of 96.50%, 95.66% and 91.06%, respectively (P<0.01). Binary logistic regression analysis result showed that different intervention strategies affected the correct use of personal RPE among dust-exposed workers after adjusting for gender, age, and other confounding factors (P<0.05). Compared with the control group, the rates of correct RPE use increased in the low-intensity intervention group and the high-intensity intervention group (odd ratio was 2.14 and 3.01; 95% confidence interval was 1.12 - 4.10 and 1.53 - 5.91, respectively). Conclusion The implementation of traditional plus mobile health education interventions on the proper use of RPE can promote correct RPE utilization among dust-exposed workers, and integrating peer education further enhances the intervention effectiveness.

Objective To explore the clinical effect of applying an improved"outside-in"technique in hip arthroscopic surgery.Methods Totally 136 patients undergoing hip arthroscopic surgery between June 2021 and July 2023 were selected and studied retrospectively.According to their different surgi-cal approach,they were divided into a modified approach group(n=75,including 30 males and 45 fe-males,with an average age of 36±14)and a classic approach group(n=61,including 33 males and 28 females,with an average age of 31±11).Before as well as 4 weeks,3 months and 6 months after the surgery,both groups were evaluated using the visual analog scale(VAS)and Harris hip score,and observed their surgical complications.Moreover,the surgical outcomes and time of sur-gical approach establishment were compared between the two groups.Results There was no significant difference between the two groups in general information,preoperative VAS and Harris scores.More-over,no significant difference was found between the two groups in the Harris score after surgery.Compared with the classic approach group,the improved group had significantly less surgical time(49.0±16.9 min vs.66.0±13.3 min,P<0.05),without using the C-arm fluoroscopy during surgery.Moreover,in the modified approach group,the time for establishing the mid-anteriorapproach was sig-nificantly shortened(10.4±5.9 min vs.25.9±15.1 min,P<0.05),along with the traction time during surgery(66.0±13.3 min vs.49.0±16.9 min,P<0.05).Conclusion The modified"outside-in"hip ar-throscopy technique is a safe and effective surgical method,easier to operate,with shorter surgical time.

Objective To observe the effects of the traditional Chinese medicine Qingfeihuayutongfu prescription on oxygenation index,respiratory mechanics,inflammatory markers,and immune function in mechanically ventilated patients with sepsis-associated acute respiratory distress syndrome(ARDS).Methods A prospective randomized controlled trial was conducted.Sixty-eight patients with sepsis-associated ARDS,who met the TCM syndrome pattern of lung-heat transferring to intestines syndrome,admitted to the department of intensive care unit(ICU)of the Affiliated Hospital of Nanjing University of Chinese Medicine between January 2023 and January 2024,were enrolled as study subjects.Patients were randomly divided into an experimental group and a control group using a random number table,with 34 patients in each group.Four patients dropped out from each group,resulting in 30 patients being analyzed in each group.Both groups received conventional Western medical treatment after admission.The experimental group additionally received Qingfeihuayutongfu prescription[composition:Scutellaria baicalensis 10 g,Mori Cortex 10 g,Descurainiae Semen 15 g,Trichosanthis Fructus 10 g,Lumbricus 10 g,Persicae Semen 10 g,Salviae Miltiorrhizae Radix 10 g,Curcumae Rhizoma 6 g,Rheum palmatum 3 g(decocted later),Aurantii Fructus Immaturus 10 g,Magnoliae Officinalis Cortex 10 g,Glycyrrhizae Radix praeparata 3 g].The decoction was concentrated to 100 mL per bag.One dose was administered daily,divided into 2 nasogastric feedings.The control group received an equivalent volume of warm water twice daily via nasogastric tube in addition to conventional Western treatment.Treatment continued for 7 days in both groups.Differences in oxygenation index,respiratory mechanics parameters,inflammatory cytokines,and immune cell levels before and after treatment were compared between the two groups.Results Prior to treatment,no statistically significant differences were observed between the two groups in oxygenation index,respiratory mechanics parameters,levels of inflammatory cytokines or immune cell counts.Following treatment,both groups exhibited a significant increase in oxygenation index and natural killer cell(NK cell),alongside significant decreases in plateau pressure(Pplat),driving pressure(ΔP),hypersensitive C-reactive protein(hs-CRP),procalcitonin(PCT),interleukins(IL-6,IL-8,IL-1β),tumour necrosis factor-α(TNF-α),and CD4+/CD8+ratio compared to baseline.Post-treatment comparison between groups revealed that the experimental group had a significantly higher oxygenation index and NK cell than the control group[oxygenation index(mmHg,1 mmHg≈0.133 kPa):331.32±90.89 vs.238.64±83.26,NK cell:0.20(0.12,0.25)vs.0.10(0.08,0.19),both P<0.05].Conversely,the experimental group demonstrated significantly lower values than the control group for Pplat,ΔP,hs-CRP,PCT,IL-6,IL-8,TNF-α,CD4+/CD8+[Pplat(cmH2O,1 cmH2O≈0.098 kPa):16(15,19)vs.22(19,24),ΔP(cmH2O):11±2 vs.14±3,hs-CRP(mg/L):21.32(11.63,31.84)vs.41.36(17.41,89.02),PCT(μg/L):0.13(0.08,0.21)vs.0.45(0.14,1.35),IL-6(ng/L):16.25(10.72,49.96)vs.66.70(25.82,195.64),IL-8(ng/L):5.48(0.84,12.60)vs.26.23(23.10,50.16),TNF-α(ng/L):0.77(0.72,1.20)vs.1.10(0.92,1.48),CD4+/CD8+:1.76(1.43,2.00)vs.2.12(1.77,4.03),all P<0.05].Conclusion The Qingfeihuayutongfu prescription can effectively improves the oxygenation index,reduces Pplat and ΔP,mitigates inflammation,and modulates immune function in mechanically ventilated patients with sepsis-associated ARDS.

Objective To investigate the mechanism by which Huangqi Guizhi Wuwu decoction(HQGZWWD)regulates nerve injury(NI)through vascular endothelial growth factor(VEGF)pathway based on a combination of network pharmacology and molecular docking techniques.Methods Active ingredients and targets of HQGZWWD were identified through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.A drug-ingredient-target network was constructed by integrated with data from OMIM,GeneCards,and CTD databases to identify VEGF/NI-related targets.Protein-protein interaction analyses were conducted by using STRING network platform,which were further analyzed by using the DAVID database for Gene Ontology/Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.A"compound-target-pathway"network was constructed by using Cytoscape.Molecular docking was performed to validate the binding ability of core components with the targets.Results The primary constituents(quercetin,kaempferol)and principal targets[signal transducer and activator of transcription 3(STAT3),caspase 3,epidermal growth factor receptor(EGFR),etc.]of HQGZWWD were identified.KEGG analysis revealed that the targets were enriched cancer,EGFR inhibitor resistance,and advanced glycation end-products-receptor for advanced glycation end-products etc.signaling pathways.Molecular docking demonstrated binding energy between core components and STAT3,caspace 3,EGFR,etc.,specifically,the core components exhibited strong binding energy such as STAT3(≤-7.37kcal/mol).Conclusion HQGZWWD shows potency in suppressing inflammation,oxidative stress,and apoptosis,while promoting neural repair through VEGF regulation.The integration of network pharmacology and molecular docking offers a foundational framework for mechanistic investigations.

AIM:To identify surrogate clinical bio-markers for profiling the eosinophilic status of an individual patient over induced sputum analysis.METHODS:We conducted a cross-sectional study on 100 asthmatic patients whose induced sputum was successfully collected.Subjects were further classified into either EA or NEA based on whether the percentage of sputum eosinophil count(SEC％)was≥3％.Demographic and clinical data were col-lected,including basic information,routine blood tests,lung function tests,bronchodilator reversibili-ty tests,fractional exhaled nitric oxide(FeNO),the Asthma Control Test(ACT)and the Asthma Control Questionnaire(ACQ).All variables significantly asso-ciated with EA were candidates for multivariate lo-gistic regression analysis.A scoring system present-ed as a nomogram for the prediction of EA was de-veloped.RESULTS:In the univariate analysis,com-pared with NEA subjects,those with EA were of older age and had worse asthma control and lung function in addition to higher values of blood eosin-ophils,serum IgE and FeNO.Multivariable logistic regression analysis revealed that age,FeNOx serum IgE and blood eosinophil count(BEC)were identi-fied as independent risk factors for eosinophilic asthma,which were all included in the nomogram.CONCLUSION:A combination assessment including age,FeNOx serum IgE and BEC could be applicable to clinicians in identifying eosinophilic asthma and is easier,faster,more inexpensive and more readily available than the induced sputum test.