Mitochondrial dysfunction in chondrocytes is a key pathogenic factor in osteoarthritis (OA), but directly modulating mitochondria in vivo remains a significant challenge. This study is the first to verify a correlation between mitochondrial dysfunction and the downregulation of the FOXO3 gene in the cartilage of OA patients, highlighting the potential for regulating mitophagy via FOXO3 gene modulation to alleviate OA. Consequently, we developed a chondrocyte-targeting CRISPR/Cas9-based FOXO3 gene-editing tool (FoxO3) and integrated it within a nanoengineered 'truck' (NETT, FoxO3-NETT). This was further encapsulated in injectable hydrogel microspheres (FoxO3-NETT@SMs) to harness the antioxidant properties of sodium alginate and the enhanced lubrication of hybrid exosomes. Collectively, these FoxO3-NETT@SMs successfully activate mitophagy and rebalance mitochondrial function in OA chondrocytes through the Foxo3 gene-modulated PINK1/Parkin pathway. As a result, FoxO3-NETT@SMs stimulate chondrocytes proliferation, migration, and ECM production in vitro, and effectively alleviate OA progression in vivo, demonstrating significant potential for clinical applications.

Accurate prediction of drug responses in cancer cell lines (CCLs) and transferable prediction of clinical drug responses using CCLs are two major tasks in personalized medicine. Despite the rapid advancements in existing computational methods for preclinical and clinical cancer drug response (CDR) prediction, challenges remain regarding the generalization of new drugs that are unseen in the training set. Herein, we propose a multimodal fusion deep learning (DL) model called drug-target and single-cell language based CDR (DTLCDR) to predict preclinical and clinical CDRs. The model integrates chemical descriptors, molecular graph representations, predicted protein target profiles of drugs, and cell line expression profiles with general knowledge from single cells. Among these features, a well-trained drug-target interaction (DTI) prediction model is used to generate target profiles of drugs, and a pretrained single-cell language model is integrated to provide general genomic knowledge. Comparison experiments on the cell line drug sensitivity dataset demonstrated that DTLCDR exhibited improved generalizability and robustness in predicting unseen drugs compared with previous state-of-the-art baseline methods. Further ablation studies verified the effectiveness of each component of our model, highlighting the significant contribution of target information to generalizability. Subsequently, the ability of DTLCDR to predict novel molecules was validated through in vitro cell experiments, demonstrating its potential for real-world applications. Moreover, DTLCDR was transferred to the clinical datasets, demonstrating satisfactory performance in the clinical data, regardless of whether the drugs were included in the cell line dataset. Overall, our results suggest that the DTLCDR is a promising tool for personalized drug discovery.

Inflammatory bowel disease（IBD）is a type of disease characterized by chronic intestinal inflammation，mainly including Crohn's disease，ulcerative colitis，and undifferentiated IBD. Its pathogenesis is very complex and involves multiple factors. In recent years，studies have shown that heat shock proteins（HSPs）and cell apoptosis play important roles in the occurrence and progression of IBD. HSPs not only participate in cellular stress response，but also play a crucial role in regulating cell apoptosis. The specific mechanism of the interaction between HSPs and apoptosis in IBD is currently unclear. This article reviews the relationship and mechanism between HSPs and apoptosis in the occurrence and development of IBD，explores the roles of HSPs and apoptosis in IBD，and proposes potential treatment strategies to provide more effective treatment options for IBD patients in the future.

Objective To investigate the molecular targets and signaling pathways involved in the therapeutic effects of Lishukang Capsule(LSK)in a rat model of high-altitude pulmonary edema(HAPE)using a proteomics-based approach.Methods A total of 60 male Wistar rats were randomly assigned to a control group,a HAPE model group,3 LSK treatment groups receiving low-,medium-,and high-dose LSK,respectively,and a Rhodiola rosea(also known as Hongjitian[HJT]in pinyin,a Chinese Romanization system)control group.After HAPE modeling,the pharmacodynamic effects were assessed and the optimal LSK dose was determined using HE stains,inflammatory cytokine quantification,lung tissue water content,and the protein concentration in bronchoalveolar lavage.Label free quantitative proteomic profiling was then applied to identify differentially expressed proteins(DEPs)in the optimal dose group,using a screening threshold of over 1.5-fold change and P<0.05.The selected DEPs were validated with Western blotting,followed by Gene Ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Results The medium-dose LSK group exhibited significant anti-HAPE effects.Findings from the proteomic analysis revealed,in the comparison with the control group,267 DEPs were identified in the HAPE group.In the comparison with the HAPE group,225 DEPs were identified in the medium-dose LSK group.A total of 112 DEPs in the control group were normalized following LSK treatment in the medium-dose LSK group.In addition,GO enrichment analysis of proteins differentially expressed between the HAPE and LSK group showed that these DEPs were mainly enriched in 12 biological processes,2 cellular components,and 5 molecular functions.KEGG pathway analysis showed that LSK activated pathways associated with cell adhesion molecules,glycosaminoglycan biosynthesis,DNA replication/nucleotide excision repair,transcriptional dysregulation in cancer,and Herpes simplex virus type 1(HSV-1)infection,while inhibiting pathways associated with glycerophospholipid metabolism.Some differentially expressed proteins with potential functions were verified by Western blotting,including AGPAT5,NCAM1,SRSF3,and PLA2.These differentially expressed proteins were significantly expressed in the normal group,HAPE group,and LSK group,and the validation results were consistent with proteomic findings,indicating the high reliability of the proteomic results.Conclusion LSK exerts a significant protective effect against HAPE.Proteomic analysis suggests that its therapeutic action may be mediated through activating pathways involved in cell adhesion molecules,glycosaminoglycan biosynthesis,DNA replication/nucleotide excision repair,transcriptional dysregulation in cancer,and HSV-1 infection,alongside inhibition of pathways associated with glycerophospholipid metabolism.The key DEPs identified in these pathways may play crucial roles in the preventive and therapeutic effects of LSK on HAPE.

Accurate prediction of drug responses in cancer cell lines(CCLs)and transferable prediction of clinical drug responses using CCLs are two major tasks in personalized medicine.Despite the rapid advancements in existing computational methods for preclinical and clinical cancer drug response(CDR)prediction,chal-lenges remain regarding the generalization of new drugs that are unseen in the training set.Herein,we propose a multimodal fusion deep learning(DL)model called drug-target and single-cell language based CDR(DTLCDR)to predict preclinical and clinical CDRs.The model integrates chemical descriptors,mo-lecular graph representations,predicted protein target profiles of drugs,and cell line expression profiles with general knowledge from single cells.Among these features,a well-trained drug-target interaction(DTI)prediction model is used to generate target profiles of drugs,and a pretrained single-cell language model is integrated to provide general genomic knowledge.Comparison experiments on the cell line drug sensitivity dataset demonstrated that DTLCDR exhibited improved generalizability and robustness in predicting unseen drugs compared with previous state-of-the-art baseline methods.Further ablation studies verified the effectiveness of each component of our model,highlighting the significant contribution of target information to generalizability.Subsequently,the ability of DTLCDR to predict novel molecules was validated through in vitro cell experiments,demonstrating its potential for real-world applications.Moreover,DTLCDR was transferred to the clinical datasets,demonstrating satisfactory performance in the clinical data,regardless of whether the drugs were included in the cell line dataset.Overall,our results suggest that the DTLCDR is a promising tool for personalized drug discovery.

Objective:To analyze the efficacy of unilateral arytenoid chondroplasty combined with minimally invasive mucosal flap plasty in the management bilateral vocal cord paralysis.Methods:A total of 66 patients with bilateral vocal cord paralysis hospitalized in the First Hospital of Sun Yat-sen University from January 2018 to December 2023 were retrospectively analyzed, among whom there were 8 males and 58 females, with ages ranging from 35 to 86 years old(mean age (57.8±11.6) years). All patients underwent suspension laryngoscopic CO 2 laser unilateral arytenoid chondroplasty and mucosal flap under general anesthesia. Postoperative follow-up period extended from 6 months to 6 years, with a median duration of 28 months. The study compared the degree of dyspnea and voice quality (subjective and objective evaluation) of the patients pre- and post-operatively, and analyzed the clinical differences between patients with and without preoperative or intraoperative tracheotomy, the extubation rate of tracheotomized patients, the recurrence rate, and the complication rate. Continuous variables conforming to normal distribution were tested by t-test and categorical variables by χ2 test,the Wilcoxon rank-sum test was used to analyze the improvement in dyspnea before and after surgery. Results:Compared with the preoperative period, 59 patients showed improvement in postoperative dyspnea ( U=161.5, P<0.01); there was no significant difference in voice disorder index 10, subjective auditory-perceptual assessment, and maximal vocalization time ( P>0.05), all of which remained within the relative normal range. Tracheotomy was performed in 32 out of 66 patients, with predominantly degree Ⅲ- dyspnea (46.9%, 15/32), including 26 patients with preoperative tracheotomy and 6 patients with intraoperative tracheotomy. Among the 34 patients who did not undergo tracheotomy, the majority presented with degree Ⅱ-dyspnea (82.4%, 28/34). All patients achieved successful extubation following surgery, with a mean median time to extubation of 1 month. Conclusions:The combination of unilateral arytenoid chondroplasty and minimally invasive mucosal flap plasty represents a refined and effective therapeutic approach for bilateral vocal cord paralysis. This minimally invasive technique has the potential to reduce the rate of tracheotomy.

Objective To evaluate the interventional effects of virtual reality(VR)on cancer patients in pain,anxiety,cognitive function,upper limb function and quality of life.Methods Randomised controlled trials(RCTs)using VR technology on cancer patients were retrieved from databases of CNKI,Wanfang Database,SinoMed,VIP,PubMed,Web of Science,Embase,PsycINFO and Cochrane Library,from the inception of the databases to March 2023.Two reviewers independently examined and screened the literatures,followed by evaluation of the quality and extraction of relevant themes.Meta-analysis was conducted by using RevMan 5.3 to analyse the acquired data.Results A total of 9 RCTs involving 727 patients were included.The meta-analysis demonstrated that VR technology alleviated pain intensity in cancer patients[SMD=-1.31,95%CI(-1.96 to-0.66),P<0.001],relieved anxiety[SMD=-0.96,95%CI(-1.62 to-0.31),P=0.004],improved cognitive function[SMD=3.37,95%CI(1.74 to 5.00),P<0.001],and improved quality of life of cancer patients[SMD=0.67,95%CI(0.38 to 0.96),P<0.001].However,it posed unclear effect on upper limb function[SMD=-0.57,95%CI(-1.40 to 0.26),P=0.18].Conclusion VR technology shows potentials in alleviation of pain,reduction of anxiety and improvement of cognitive function and the quality of life in cancer patients,but it remains uncertain effect on upper limb function.However,due to heterogeneity in some of the results,more high-quality studies are required to validate the interventional effects of VR technology on cancer patients.

Objective To investigate the effect and mechanism of long chain non-coding RNA(ln-cRNA)SNHG16 regulating PAR1 on the proliferation,migration and invasion of lung cancer.Methods From March 2020 to August 2021,lung cancer tissues and adjacent tissues of 35 patients with lung cancer were col-lected,and lung cancer cell lines(HCC827,A549,SK-LU-1,A427)and normal lung cell lines(MRC5)were simultaneously cultured.The overexpression model of PAR1(pcDNA-PAR1)was constructed by using the expression vector pcDNA3.1.A549 cells were divided into four groups after transfection:si-SNHG16,si-PAR1,si-SNHG16+pcDNA-PAR1 and control(transfection with si-NC).The expression levels of lncRNA SNHG16 and PAR1 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427),lung cancer tissues and adja-cent tissues were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reac-tion(qRT-PCR),and the transfection efficiency of each group was verified.MTT assay and clonal formation were used to determine the proliferation of cells in each group,flow cytometry was used to detect the apopto-sis of cells in each group,cell scratch and Transwell test were used to detect the migration and invasion ability of cells in each group,and Western blot was used to detect the protein expression of PAR1.Results The ex-pression level of lncRNA SNHG16 in lung cancer tissue was higher than that in adjacent tissues(P＜0.05).The expression level of lncRNA SNHG16 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427)was higher than that in normal lung cell lines(MRC5),with statistical significance(P＜0.05).The results of qRT-PCR showed that the expression level of lncRNA SNHG16 gene was(21.02±0.04)％of the control af-ter transfection of si-SNHG16,the expression level of PAR1 gene was(19.06±0.02)％of the control after transfection of si-PAR1,and the expression level of PAR1 gene was 2.70±0.00 folds of the control after transfection of pcDNA-PAR 1,the difference was statistically significant(P＜0.05).The results of Western blot showed that the expression level of transfected in each PAR1 protein was different from that of the con-trol(P＜0.05).Compared with the control,the activity of cells transfected with si-SNHG16 was lower,the a-bility of clone formation,cell migration and invasion was obviously inhibited,and the apoptosis rate was higher(P＜0.05),while pcDNA-PAR1 could weaken the influence of transfected si-SNHG16 on cell proliferation,apoptosis,migration and invasion(P＜0.05).lncRNA SNHG16 was positively correlated with the expression level of PAR1(r=0.61).Conclusion lncRNA SNHG16 can regulate the occurrence and development of lung cancer by targeting PAR1.

Objective:To investigate the pathogenic mechanism and clinical characteristics of the novel splicing variant of ATP-binding cassette subfamily B member 4 (ABCB4) and provide a basis for subsequent genetic diagnosis.Methods:The clinical data of a 5-year-old child with cholestatic liver disease admitted to the Beijing Children′s Hospital of Capital Medical University was retrospectively analyzed. The pathogenic variations were detected by whole exome sequencing and verified by Sanger sequencing, and bioinformatics was used to predict the pathogenicity of the mutation sites. Possible pathogenic variations were verified in vitro by Minigene assay. The clinical outcome was followed after discharge from hospital.Results:The 5-year-old boy had developed cholestasis at the age of 11 months. His physical examination showed obvious enlargement of the liver and spleen. Cholestatic cirrhosis was diagnosed by liver function tests, abdominal ultrasonography, liver biopsy and pathology. The results of genetic analysis showed that the patient was a complex heterozygote of the ABCB4 gene, with a pathogenic mutation c.2860G>A and a novel mutation c.2065-8T>G, derived from the mother and father respectively. The conservative prediction of the c.2065-8T>G site showed that this region was highly conserved and may affect splicing. Minigene assay results confirmed that the c.2065-8T>G mutation resulted in a 7 bp retention of intron 16 in the mature mRNA. In the absence of nonsense-mediated mRNA decay, the amino acid frameshift forms a truncated protein, which is represented by p.Glu689ValfsTer19. The patient was diagnosed as progressive familial intrahepatic cholestasis type 3 (PFIC3) and treated with ursodeoxycholic acid (UDCA). His clinical symptoms improved during 18 months of follow-up.Conclusions:The c.2065-8T>G variant is confirmed to affect the splicing process and exhibits complex heterozygosity with c.2860G>A, which is identified as the cause of the disease. PFIC3 children with this variant showed cholestatic liver disease as the main manifestation with a slow progression and was sensitive to treatment with UDCA.

Objective:To explore the efficacy and safety of dual targeted therapy（DTT）in the treatment of refractory inflammatory bowel disease（IBD）in children.Methods:The diagnosis and treatment processes of refractory IBD children treated with DTT in the Department of Gastroenterology,Beijing Children's Hospital Affiliated to Capital Medical University from April 2022 to May 2024 were analyzed retrospectively,and their clinical characteristics were summarized.Results:A total of ten children with refractory IBD were included,including five males and five females,with a median onset age of 12.58 (5.25,13.33) years old,and seven cases of Crohn's disease (CD) and three cases of ulcerative colitis (UC),with a median disease course of 1.25 (0.91,4.00) years，were treated with DTT for a median time of 6.08 (6.00,13.40)months.Two (2/7) cases of CD patients were partially effective with infliximab(IFX) treatment,while five (5/7) cases were partially effective with a combination of IFX and azathioprine treatment.These patients received DTT with UST in addition to continuous using of IFX.When the seven CD patients were treated with DTT for four weeks,12 weeks and 24 weeks,and the clinical remission rates were 42.9%(3/7),71.4%(5/7) and 100%(7/7)，respectively.The pediatric Crohn's disease activity index gradually decreased,which were significantly lower than those before treatment( P<0.05).Fecal calprotectin,C-reactive protein,erythrocyte sedimentation rate,and blood white blood cells all gradually decreased,which were lower than those before treatment,while hemoglobin and serum albumin were higher than those before treatment.Three children with UC were all resistant to glucocorticoids.One case was partially effective with adalimumab treatment,one case was partially effective with IFX combined with immunosuppressive agents,and one case was partially effective with vedolizumab.All the three UC patients received DTT treatment with UST in addition to continuous using of the original biological preparation.When the three UC patients were treated with DTT for four weeks and 12 weeks,one case showed clinical remission,one case showed clinical response,and one case showed no clinical response.After 24 weeks of DTT treatment,two cases achieved clinical remission and one case had no clinical response.Re-examination of colonoscopy showed that one case of clinical remission had mucosal healing and one case had mild disease activity.However,there was a slight improvement in disease activity under endoscopy compared with that before DTT in patient who had no clinical response.During DTT treatment,no adverse events occurred in all patients. Conclusion:DTT is effective and relatively safe for children with refractory IBD,and can be one of the attempts for children with IBD when they are partially effective with one biological agent.