Antibodies play a critical role in adaptive immune responses and serve as key components in disease diagnosis and treatment. These molecules exhibit dynamic post-translational modifications (PTMs), such as glycosylation and phosphorylation, which regulate their effector functions. To date, nearly all of our knowledge about antibody repertoires has come from B cell receptor (BCR) sequencing (BCR-seq), which facilitates the profiling of clonal composition and the tracing of maturation trajectories within B-cell repertoires. However, circulating antibodies found in bodily fluids—such as serum, saliva, milk, mucosal secretions, and cerebrospinal fluid—exhibit diversities and specificities beyond what BCR-seq alone can predict. Therefore, identifying and quantifying antibody clonotypes at the protein level could enhance diagnosis, prognosis, and treatment strategies in personalized medicine. The critical gap between genotype and phenotype necessitates complementary methodologies that enable the direct characterization of antibody proteins in their native functional states. Mass spectrometry (MS)-based antibody repertoire sequencing (Ab-seq) is currently the only feasible approach for this task and primarily includes database-dependent methods—such as bottom-up, middle-down, and top-down approaches—as well as database-independent de novo sequencing technology. These strategies enable multi-level, high-precision characterization ranging from peptides and domains to intact antibody molecules. Unlike the shotgun strategy commonly used in routine proteomics, obtaining full sequences of all antibodies presents unique challenges. It requires specialized methodological adaptations to address issues related to dynamic range, sequence variation, and sample complexity. This review introduces the technical principles, methodological workflows, and recent applications of various mass spectrometry-based antibody repertoire sequencing (Ab-seq) strategies, with a focus on approaches designed to improve sequence coverage and identification accuracy. These include multi-enzyme digestion, hybrid fragmentation methods, and artificial intelligence-assisted de novo sequencing. By systematically comparing database-dependent techniques—such as bottom-up, middle-down, and top-down approaches—with database-independent de novo sequencing, this review outlines their respective advantages and limitations in terms of sample throughput, sequence coverage, post-translational modification characterization, and data analysis complexity. In addition, this review discusses emerging technological trends, including the integration of ion mobility separation, native mass spectrometry, and artificial intelligence-driven data interpretation, which are expected to enhance the depth and accuracy of antibody characterization. Although current methods continue to face challenges related to sample complexity, dynamic range, and unambiguous sequence variant assignment, we emphasize the importance of integrating BCR-seq and Ab-seq data to construct gene-protein association maps. These maps help validate sequence accuracy and facilitate epitope discovery. This dual-platform strategy helps bridge the gap between genotype and phenotype, thereby enhancing both the resolution and scope of antibody repertoire studies. Such an integrative approach also offers a valuable tool for therapeutic antibody development, structure-function analysis, and precise evaluation of vaccine efficacy.

Objective:To summarize the clinical data of single-center juvenile dermatomyositis（JDM）complicated with interstitial lung disease（ILD），and provide experience for pediatricians.Methods:Data of 61 children with JDM who were admitted to Children's Hospital affiliated to Xi'an Jiaotong University from January 2016 to May 2023 were collected. General data，clinical symptoms，chest high-resolution CT，laboratory examination and myositis antibody spectrum of the children were recorded.Results:Among the 61 children with JDM，there were 30 cases（13 males and 17 females）without ILD. The age of onset was 5.96（3.50，8.92）years and the course of disease was（11.79±20.00）months. There were 31 cases with ILD（14 males and 17 females），the age of onset was 7.42（4.50，10.08）years，and the duration of ILD was（5.47±8.09）months. There was statistical difference in the course of disease between the two groups（ P<0.05），but no statistical difference in gender and age between the two groups（ P>0.05）. Among 61 children with JDM，there were statistical differences in fever between the two groups（ P<0.05），but no statistical differences in heliotrope discoloration，gottron’s papules，calcinosis and myasthenia between the two groups（ P>0.05）. AST and FER showed statistical difference between the two groups（ P<0.05），while CK，LDH，CK-MB，ESR，C3 and C4 showed no statistical difference（ P>0.05）. All 61 cases of children were tested for myositis antibody spectrum，and there was statistical difference in anti-MDA5 antibody between the two groups（ P<0.05），but no statistical difference in the rest（ P>0.05）. There were statistical differences between the two groups in the treatment of methotrexate，hydroxychloroquine and cyclophosphamide（ P<0.05）. A total of 11 cases（36.67%）in the without ILD group were treated with biologics（8 adalimumab，2 infliximab and 1 tofacitinib），and 23 cases（74.19%）in the ILD group were treated with biologics（11 adalimumab，9 tofaciib，2 infliximab and 1 tocilizumab）. All 61 cases with JDM were followed up. Among the 30 children without ILD，1 case was lost to follow-up 2 months after treatment，and the rest were treated effectively without death. Among the 31 children with ILD，3 cases died of severe pulmonary infection with multidrug-resistant bacteria during treatment，of which 1 case was positive for anti-MDA5 antibody and 2 cases were negative for myositis specific antibody. Conclusion:JDM is more likely to be complicated with ILD，fever is more likely to occur in ILD group，and children with positive anti-MDA5 antibody are more likely to occur ILD. Biologic agents such as adalimumab and tofacitinib are effective in combination therapy. In the course of treatment，multi-drug resistant bacteria infection should be guarded against to reduce mortality.

Objective:To evaluate the efficacy of 3D printing technology combined with computer navigation-assisted screw implantation in the treatment of atlantoaxial instability (AAI) complicated by vertebral artery anomalies.Methods:A retrospective case series study was conducted to analyze the clinical data of 23 patients with AAI complicated by vertebral artery anomalies who were admitted to Honghui Hospital of Xi′an Jiaotong University between January 2019 and January 2023, including 10 males and 13 females, aged 19-70 years [(51.0±13.3)years]. Vertebral artery anomalies were categorized into unilateral high-riding vertebral artery with unilateral dominance ( n=14), bilateral high-riding vertebral arteries with unilateral dominance ( n=6), and ponticulus posticus ( n=3). All the patients underwent preoperative planning using a 3D-printed model of the atlantoaxial complex with the vertebral artery, followed by posterior atlantoaxial pedicle screw fixation with computer-assisted navigation. Operative duration and intraoperative blood loss were recorded. The accuracy of pedicle screw placement was assessed at 3 days postoperatively using the Gertzbein-Robbins classification. Visual analogue scale (VAS) scores and Japanese Orthopedic Association (JOA) scores were evaluated preoperatively, at 3 days, 3 months postoperatively, and at the last follow-up. Bony fusion was assessed using cervical CT with 3D reconstruction at the last follow-up. Complications were also observed. Results:All the patients were followed up for 12-19 months [(15.1±1.9)months]. The operative duration was 125-167 minutes [(140.6±10.9)minutes] and intraoperative blood loss was 200-600 ml [(295.7±77.8)ml]. At 3 days postoperatively, all the 66 pedicle screws were safely placed, with 60 screws (91%) rated as Gertzbein-Robbins Grade 0 and 6 screws (9%) as Grade 1. At 3 days and 3 months postoperatively, and at the last follow-up, the VAS scores were (4.0±1.0)points, (2.0±0.6)points, and (1.3±0.5)points, and the JOA scores were (14.2±1.2)points, (16.0±0.8)points, and (16.6±0.5)points, both of which were not only significantly improved compared with preoperative (5.6±1.3)points and (12.8±1.5)points, but also further improved over time ( P<0.05). At the last follow-up, 22 patients (96%) achieved satisfactory atlantoaxial bony fusion. No vertebral artery injury, spinal cord or nerve injury, cerebrospinal fluid leakage, or screw loosening were observed in any patients. Conclusion:For patients with AAI complicated by vertebral artery anomalies, 3D printing combined with computer navigation-assisted navigation for atlantoaxial pedicle screw implantation offers multiple advantages, including minimal surgical trauma, high screw placement accuracy, pain relief, neurological function improvement, high fusion rate, and lowered incidence of complications.

Objective To investigate the characteristics of macrophage depletion by clodronate liposomes(CL)in a carbon tetrachloride(CCl4)-induced liver fibrosis mouse model,and to analyze the transcriptomic features.Methods Thirty-two C57BL/6 mice were divided randomly into plain control liposomes for clophosome(PL)and clodronate liposome(CL)groups(n=16 mice per group),and administered intraperitoneal injections of PL and CL,respectively.On day 5,each group was further divided into normal(N)and model(M)subgroups(n=8 mice per subgroup).Mice in group M received 10％CCl4 intraperitoneally to induce liver fibrosis,while mice in group N received an equal volume of olive oil.After 4 weeks,serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels were measured,and hepatic inflammation and collagen deposition were evaluated by hematoxylin/eosin and Sirius red staining,respectively.Total RNA was extracted from liver tissues for transcriptomic sequencing and subsequent differential gene expression analysis.Results Serum ALT and AST levels were significantly elevated in the PL-M group(P＜0.01),with fibrosis staging primarily at S3,compared with S1 in the CL-M group.Totals of 1462 and 2119 differentially expressed genes(|log fold change|＞2 and P＜0.05)were identified in the PL and CL groups,respectively.Gene Ontology analysis revealed enrichment in multiple biological processes,cellular components,and molecular functions in both models,and Kyoto Encyclopedia of Genes and Genomes analysis identified 29 significantly enriched pathways(P＜0.05).The upregulation of genes including Lgals7 and Timp1 and the downregulation of Mup-ps16 and Mup15 were validated by reverse transcription-quantitative polymerase chain reaction,consistent with transcriptomic trends(P＜0.05).Conclusions This study highlights the characteristics and transcriptomic features of macrophage depletion in the CCl4-induced liver fibrosis model,providing a theoretical reference for research on the immune mechanisms of liver fibrosis.

Objective To evaluate the longitudinal imaging features of coronary artery plaques in elderly male patients with CHD and myocardial bridges and explore the longitudinal changing pat-terns.Methods A total of 117 elderly male CHD patients who underwent two examinations of coronary computed tomography angiography in our medical center from January 2018 to Septem-ber 2023 were enrolled in this study.Then 216 small plaques(0.1-50 mm3 in size)were subjec-ted,and classified into the proximal myocardial bridge group(98 plaques)and other heart part group(118 plaques)according to the site of the plaques.Plaque volume,plaque composition vol-ume,FAI,and CT-derived fraction flow reserve(CT-FFR)were calculated and recorded.Results In the 2 groups of plaques,there were no statistically differences in the plaque length,plaque nec-rotic core volume,and FAI derived from the second examination than the baseline one(P＞0.05).The plaque volume,intra-plaque fibers,and dense calcified volume of plaques in the second exami-nation were significantly greater than those at baseline,and CT-FFR was obviously smaller than the baseline level in both groups(P＜0.05,P＜0.01).In the proximal myocardial bridge group,the intra-plaque fibrofat volume in the second examination was significantly larger than that of baseline,while opposite phenomenon was observed in the plaques of the other heart part group(P＜0.05).The annual changing rates of intraplaque fibrofat volume and FAI were significantly higher in the proximal myocardial bridge group than the other heart part group[0.51％(-0.32％,0.51％)vs 0.02％(-0.46％,0.20％),P=0.046;0.55％(-2.44％,1.76％)vs 0.33％(-1.36％,2.63％),P=0.044].Conclusion In elderly male patients,the intraplaque fibrofat vol-ume,FAI and CT-FFR are more likely to change in the proximal plaques of the left anterior de-scending artery myocardial bridge than the plaques of other parts of heart,so the proximal plaques of the left anterior descending artery need more clinical attention and early intervention.

Objective To analyze the epidemiological characteristics of human papillomavirus(HPV)infection in women in the Zhongguancun area of Beijing,and to evaluate the infection rates and genotype distribution between different populations and age groups.Methods A retrospective analysis was conducted on HPV genotyping results of 13,105 women who visited the gynecology outpatient department or underwent routine health check-ups at Zhongguancun Hospital from March 2019 to April 2024.High-risk HPV genotypes(15 types)were detected using a fluorescence PCR assay.Positive cases were classified as single,dual or multiple(≥3)infections based on the number of genotypes.Subjects were stratified into six age groups(≤30,31-40,41-50,51-60,61-70,and≥71 years),and the characteristics of infection by type and age group were analyzed.Results The overall HPV positivity rate was 10.78%(1,413/13,105),with a significantly higher rate in the outpatient group than that in the health check-up group(16.36%vs.6.06%,P<0.01).The three most prevalent genotypes were HPV52(18.83%),HPV58(13.85%),and HPV16(11.28%).Single infections accounted for 79.19%of cases,dual infections for 15.93%and multiple infections for 4.88%.Age distribution showed a U-shaped pattern,with higher infection rate in women aged≤30 years(15.06%)and 61-70 years(13.19%),and the lowest rate in the≥71 years(8.09%).Notably,women aged≤30 years had the highest proportion of multiple infections(31.72%).Conclusion These findings provide a basis for cervical cancer screening strategies,HPV vaccination promotion and individualized prevention of cervical cancer in this region.

Objective:To evaluate the efficacy of 3D printing technology combined with computer navigation-assisted screw implantation in the treatment of atlantoaxial instability (AAI) complicated by vertebral artery anomalies.Methods:A retrospective case series study was conducted to analyze the clinical data of 23 patients with AAI complicated by vertebral artery anomalies who were admitted to Honghui Hospital of Xi′an Jiaotong University between January 2019 and January 2023, including 10 males and 13 females, aged 19-70 years [(51.0±13.3)years]. Vertebral artery anomalies were categorized into unilateral high-riding vertebral artery with unilateral dominance ( n=14), bilateral high-riding vertebral arteries with unilateral dominance ( n=6), and ponticulus posticus ( n=3). All the patients underwent preoperative planning using a 3D-printed model of the atlantoaxial complex with the vertebral artery, followed by posterior atlantoaxial pedicle screw fixation with computer-assisted navigation. Operative duration and intraoperative blood loss were recorded. The accuracy of pedicle screw placement was assessed at 3 days postoperatively using the Gertzbein-Robbins classification. Visual analogue scale (VAS) scores and Japanese Orthopedic Association (JOA) scores were evaluated preoperatively, at 3 days, 3 months postoperatively, and at the last follow-up. Bony fusion was assessed using cervical CT with 3D reconstruction at the last follow-up. Complications were also observed. Results:All the patients were followed up for 12-19 months [(15.1±1.9)months]. The operative duration was 125-167 minutes [(140.6±10.9)minutes] and intraoperative blood loss was 200-600 ml [(295.7±77.8)ml]. At 3 days postoperatively, all the 66 pedicle screws were safely placed, with 60 screws (91%) rated as Gertzbein-Robbins Grade 0 and 6 screws (9%) as Grade 1. At 3 days and 3 months postoperatively, and at the last follow-up, the VAS scores were (4.0±1.0)points, (2.0±0.6)points, and (1.3±0.5)points, and the JOA scores were (14.2±1.2)points, (16.0±0.8)points, and (16.6±0.5)points, both of which were not only significantly improved compared with preoperative (5.6±1.3)points and (12.8±1.5)points, but also further improved over time ( P<0.05). At the last follow-up, 22 patients (96%) achieved satisfactory atlantoaxial bony fusion. No vertebral artery injury, spinal cord or nerve injury, cerebrospinal fluid leakage, or screw loosening were observed in any patients. Conclusion:For patients with AAI complicated by vertebral artery anomalies, 3D printing combined with computer navigation-assisted navigation for atlantoaxial pedicle screw implantation offers multiple advantages, including minimal surgical trauma, high screw placement accuracy, pain relief, neurological function improvement, high fusion rate, and lowered incidence of complications.

Objective To evaluate the longitudinal imaging features of coronary artery plaques in elderly male patients with CHD and myocardial bridges and explore the longitudinal changing pat-terns.Methods A total of 117 elderly male CHD patients who underwent two examinations of coronary computed tomography angiography in our medical center from January 2018 to Septem-ber 2023 were enrolled in this study.Then 216 small plaques(0.1-50 mm3 in size)were subjec-ted,and classified into the proximal myocardial bridge group(98 plaques)and other heart part group(118 plaques)according to the site of the plaques.Plaque volume,plaque composition vol-ume,FAI,and CT-derived fraction flow reserve(CT-FFR)were calculated and recorded.Results In the 2 groups of plaques,there were no statistically differences in the plaque length,plaque nec-rotic core volume,and FAI derived from the second examination than the baseline one(P＞0.05).The plaque volume,intra-plaque fibers,and dense calcified volume of plaques in the second exami-nation were significantly greater than those at baseline,and CT-FFR was obviously smaller than the baseline level in both groups(P＜0.05,P＜0.01).In the proximal myocardial bridge group,the intra-plaque fibrofat volume in the second examination was significantly larger than that of baseline,while opposite phenomenon was observed in the plaques of the other heart part group(P＜0.05).The annual changing rates of intraplaque fibrofat volume and FAI were significantly higher in the proximal myocardial bridge group than the other heart part group[0.51％(-0.32％,0.51％)vs 0.02％(-0.46％,0.20％),P=0.046;0.55％(-2.44％,1.76％)vs 0.33％(-1.36％,2.63％),P=0.044].Conclusion In elderly male patients,the intraplaque fibrofat vol-ume,FAI and CT-FFR are more likely to change in the proximal plaques of the left anterior de-scending artery myocardial bridge than the plaques of other parts of heart,so the proximal plaques of the left anterior descending artery need more clinical attention and early intervention.

Objective To investigate the effects and mechanisms of high-fat diet(HFD)-induced obesity on male ze-brafish reproductive function.Methods Adult male zebrafish were divided into normal diet(ND)group and HFD group.Growth and metabolic conditions were evaluated by measuring body weight,body length,BMI,organ index,and glucose/lipid lev-els.Reproductive capacity was assessed via sperm concentration,motility,fertilization rate,and testosterone levels.Testicular tissues from both of groups were subjected to transeriptomic sequencing(RNA-seq).And qRT-PCR was used to validate the expression of genes.Results Compared to male zebrafish in ND group,the ones in HFD group exhibited hepatic steatosis and glucose/lipid meta-bolic disorders(P＜0.05).Testicular structural disorganization,along with reduced testosterone levels,decreased gonadosomatic in-dex,and impaired sperm concentration and motility occurred in HFD group(P＜0.05).GO analysis revealed that differentially ex-pressed genes were enriched in spermatogenesis and ciliary system,while KEGG analysis highlighted metabolic related pathways(pu-rine metabolism,thyroid hormone synthesis,mTOR signaling)and cell adhesion molecules.Twenty key differentially expressed genes were validated by qRT-PCR,which confirmed the reliability of RNA-Seq results.Conclusion Impairment of reproductive function induced by HFD in zebrafish may be associated with three regulatory mechanisms including ciliary system,metabolic dysregulation,and aberrant cell adhesion molecule signaling.This study provides mechanistic insights and identifies potential therapeutic targets for clinical management of diet-associated infertility.

Patient-Reported Outcomes(PRO)is a subjective outcome indicator based on the concept of"patient-centered",in which patients report their own information such as health status,functional status,and treatment feelings,which is an ef-fective supplement to traditional objective outcome indicators.Based on the development status of the Patient Reported Outcome Measure(PROM),it systematically analyzed the concepts related to PRO with the goal of clarifying the Patient Experience Data(PED)with research value,and proposed six core types of PROs in the medical field,including health-related quality of life,health preference,health behavior,symptom burden,functional status and patient-reported experience.For each type of PRO,the key characteristics of the study were analyzed,the evidence and application directions of related PED and the production based on PED were described,and the existing international mainstream PROMs were reviewed.It is hoped that the connotation of PRO-related concepts can be comprehensively expounded,and PEDs with research significance and value can be explored,so as to provide guidance for the focus of PRO-related research in China,so as to promote the application and development of PRO in China.