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 investigate the correlation between postoperative perceived stress,quality of life and fear of cancer recurrence(FCR)in elderly non-small-cell lung cancer(NSCLC)patients.Methods:This study was a prospective research design,a total of 147 elderly NSCLC patients who visited Shanxi Integrated Traditional Chinese and Western Medicine Hospital from January 2023 to June 2024 were prospectively selected for the study.All patients underwent thoracoscopic lobectomy.FCR in patients was evaluated by fear of disease progression simplified scale(FoP-Q-SF),and they were divided into dysregulation group and non dysregulation group according to the FoP-Q-SF score.Quality of life was evaluated by functional assessment of cancer therapy-lung cancer(FACT-L).Perceived stress was evaluated by Chinese version of the perceived stress scale(CPSS)score.The scores of postoperative perceived stress,quality of life and postoperative FCR in elderly NSCLC patients were analyzed,and the influence of demographic and clinical characteristics on the postoperative FCR in elderly NSCLC patients.The postoperative perceived stress and quality of life scores of elderly NSCLC patients with different FCR were compared,the correlation between postoperative FCR and perceived stress and quality of life in elderly NSCLC patients were analyzed,as well as the model fitting index of the influence of postoperative perceived stress and quality of life on FCR in elderly NSCLC patients.Results:The postoperative CPSS score in elderly NSCLC patients was(52.17±4.02)points,the FACT-L score was(80.39±4.56)points,and the total score of FoP-Q-SF was(31.16±8.69)points.Family relationship,economic burden of disease,chemotherapy and clinical stage were closely associated with the postoperative FCR in elderly NSCLC patients(P＜0.05).The CPSS score in dysregulation group was higher than that in nondysregulation group,and the FACT-L score was lower than that in nondysregulation group(P＜0.05).Pearson correlation analysis,postoperative FCR in elderly NSCLC patients was positively correlated with perceived stress,but negatively correlated with quality of life(P＜0.05).The postoperative quality of life in elderly NSCLC patients was recorded as the independent variable,the postoperative perceived stress was recorded as the mediating variable,and the postoperative FCR was recorded as the dependent variable,a structural model of the mediating effect was routinely constructed.The results showed that the model fits well.Quality of life had a direct negative predictive effect on postoperative FCR in elderly NSCLC patients(P＜0.05),and could affect FCR through postoperative perceived pressure(P＜0.05),with the mediating effect accounting for 34.15％of the total effect.After repeatedly sampling the data using the Bootstrap method and implementing the mediating effect,the 95％CI of each model path did not include 0,indicating that the difference in the mediating effect was statistically significant(P＜0.05).Conclusion:Postoperative FCR in elderly NSCLC patients is positively correlated with perceived stress,but negatively correlated with quality of life,clinically,monitoring such indicators can be considered to better evaluate the postoperative FCR in elderly NSCLC patients.

Ventricular assist devices(VADs)have emerged as a pivotal therapeutic option for end-stage heart failure,undergoing significant technological advancements from pneumatic membrane pumps to continuous-flow centrifugal pumps.This review provides a detailed overview of several VAD models,including HeartMate Ⅲ,EVAHEART 2,and Heart Con.The initial generation of VADs established a foundation for subsequent developments,despite the occurrence of frequent complications.The second generation of VADs demonstrated a notable improvement in patient prognosis but carried the risk of ventricular collapse.The third generation of VADs represents a further reduction in size,incorporating magnetic levitation technology to enhance durability and blood compatibility.In the future,the development of VADs will focus on better emulating the physiological function of the native heart,improving pulsatility,and extending device longevity.As technology advances,VADs are expected to offer heart failure patients with a greater range of treatment options and a higher quality of life.

Objective:To investigate the correlation between postoperative perceived stress,quality of life and fear of cancer recurrence(FCR)in elderly non-small-cell lung cancer(NSCLC)patients.Methods:This study was a prospective research design,a total of 147 elderly NSCLC patients who visited Shanxi Integrated Traditional Chinese and Western Medicine Hospital from January 2023 to June 2024 were prospectively selected for the study.All patients underwent thoracoscopic lobectomy.FCR in patients was evaluated by fear of disease progression simplified scale(FoP-Q-SF),and they were divided into dysregulation group and non dysregulation group according to the FoP-Q-SF score.Quality of life was evaluated by functional assessment of cancer therapy-lung cancer(FACT-L).Perceived stress was evaluated by Chinese version of the perceived stress scale(CPSS)score.The scores of postoperative perceived stress,quality of life and postoperative FCR in elderly NSCLC patients were analyzed,and the influence of demographic and clinical characteristics on the postoperative FCR in elderly NSCLC patients.The postoperative perceived stress and quality of life scores of elderly NSCLC patients with different FCR were compared,the correlation between postoperative FCR and perceived stress and quality of life in elderly NSCLC patients were analyzed,as well as the model fitting index of the influence of postoperative perceived stress and quality of life on FCR in elderly NSCLC patients.Results:The postoperative CPSS score in elderly NSCLC patients was(52.17±4.02)points,the FACT-L score was(80.39±4.56)points,and the total score of FoP-Q-SF was(31.16±8.69)points.Family relationship,economic burden of disease,chemotherapy and clinical stage were closely associated with the postoperative FCR in elderly NSCLC patients(P＜0.05).The CPSS score in dysregulation group was higher than that in nondysregulation group,and the FACT-L score was lower than that in nondysregulation group(P＜0.05).Pearson correlation analysis,postoperative FCR in elderly NSCLC patients was positively correlated with perceived stress,but negatively correlated with quality of life(P＜0.05).The postoperative quality of life in elderly NSCLC patients was recorded as the independent variable,the postoperative perceived stress was recorded as the mediating variable,and the postoperative FCR was recorded as the dependent variable,a structural model of the mediating effect was routinely constructed.The results showed that the model fits well.Quality of life had a direct negative predictive effect on postoperative FCR in elderly NSCLC patients(P＜0.05),and could affect FCR through postoperative perceived pressure(P＜0.05),with the mediating effect accounting for 34.15％of the total effect.After repeatedly sampling the data using the Bootstrap method and implementing the mediating effect,the 95％CI of each model path did not include 0,indicating that the difference in the mediating effect was statistically significant(P＜0.05).Conclusion:Postoperative FCR in elderly NSCLC patients is positively correlated with perceived stress,but negatively correlated with quality of life,clinically,monitoring such indicators can be considered to better evaluate the postoperative FCR in elderly NSCLC patients.

Objective:To investigate the clinicopathological features, genetic characteristics, and differential diagnosis of glomangiomatosis with uncertain malignant potential.Methods:Two cases of glomangiomatosis with uncertain malignant potential were collected at Henan Provincial People′s Hospital from 2013 and 2023. Immunohistochemistry and next generation sequencing (DNA-seq) were used to detect the related protein and gene variation. Patients were followed up.Results:Case 1 was male, 34 years old; and case 2 was female, 28 years old. Both had tumor recurrence in the original site. There were multiple nodules at right calf and ankle, involving superficial subcutaneous tissue and deep interfascicular muscles; some nodules were borderless and painful. Microscopically, the tumor was nodular with fibrous pseudocapsule, some had indistinct borders and diffuse infiltration to the surrounding adipose tissue. The tumor cells were round to ovoid with inconspicuous nucleoli, partly surrounding small irregularly dilated thin-walled blood vessels. The recurrent tumors showed epithelioid morphology in some of the tumor cells, with eosinophilic cytoplasm, some apparent nucleoli, mild to moderate nuclear atypia, and brisk mitotic figures. Focally, perimuscular cell differentiation was noted. The small lesion showed intravascular tumor thrombus. NGS revealed BRAF V600E mutation in case 1, and BRAF V600E mutation combined with PDGFRB gene amplification in case 2.Conclusions:Glomangiomatosis with uncertain malignant potential is a rare variant of glomus tumor. It has a unique growth pattern morphologically, BRAF V600E mutation, and invasive biological behavior.

Objective:To investigate the relationship between collateral circulation and viable myocardium (VM) in patients with coronary chronic total occlusion (CTO).Methods:A total of 88 patients (76 males, 12 females, age (61.0±9.8) years) with coronary CTO were retrospectively analyzed. All patients underwent both 99Tc m-methoxyisobutylisonitrile (MIBI) SPECT myocardial perfusion imaging and 18F-FDG PET myocardial metabolism imaging for evaluation of VM at the First People′s Hospital of Changzhou between September 2012 and June 2023, and they were scheduled to receive coronary revascularization. The perfusion/metabolism mismatch myocardium was regarded as VM. The VM index within the CTO region was calculated, reflected the quantities of VM: VM index=(summed rest score within the CTO region-summed 18F-FDG uptake score within the CTO region)/reduced perfusion myocardial segments×4×100%. Rentrop grading of collateral circulation was performed based on coronary angiography. The differences of VM index within the CTO region between poor-developed (PD, Rentrop grade 0-1) and well-developed (WD, Rentrop grade 2-3) collateral circulation, and among different Rentrop grades were analyzed by the independent-sample t test or Kruskal-Wallis rank sum test. The linear regression analysis was used to evaluate the relationship between Rentrop grading and VM index within the CTO region. The ROC curve was constructed to analyze the predictive value of Rentrop grading for VM within the CTO region. Results:The VM index within the CTO region was significantly higher in WD patients ( n=54) compared to those in PD patients ( n=34): (45.8±16.3)% vs (21.3±16.7)% ( t=-6.79, P<0.001). Moreover, the VM index within the CTO region increased with increased Rentrop grade, and there was a significant difference among 4 groups ( H=30.22, P<0.001). Multiple linear regression analysis showed that only the Rentrop grading was an independent influencing factor for the VM index within the CTO region ( β=9.29, 95% CI: 5.91-12.67, P<0.001). ROC curve showed that the sensitivity and specificity of Rentrop grading score≥2 for predicting the presence or absence of VM within the CTO region were 65.8%(52/79) and 7/9, with the AUC of 0.724(95% CI: 0.619-0.814). Conclusions:In CTO patients who are scheduled for revascularization and evaluation of VM, as the Rentrop grading increases, the VM index within the CTO region also increases. The presence of VM within the CTO region can be predicted with Rentrop grading score ≥2.

Objective:To investigate the clinical characteristics and genetic etiology of Duchenne muscular dystrophy (DMD) with myogenic tumors.Methods:The clinical data of 2 children with DMD combined with myogenic tumors diagnosed in Children′s Hospital Affiliated to Zhengzhou University in July 2021 and February 2022 were collected. The relevant literature was reviewed to summarize the clinical characteristics and explore the mechanism of the dystrophin ( DMD) gene in myogenic tumors. Results:A 6-year and 10-month-old boy with DMD (deletion of exon 45) and a 12-year-old boy with DMD (deletion of exon 51) were diagnosed with tumors. They were diagnosed with DMD for delayed motor development in the Department of Neurology of Children′s Hospital Affiliated to Zhengzhou University. They presented with painless masses in the waist. Postoperative pathological diagnosis: the pathology and immunohistochemistry of case 1 showed an alveolar rhabdomyosarcoma (ARMS) and both myogenin and myogenic differentiation 1 positive; the pathology and immunohistochemistry of case 2 showed an alveolar soft part sarcoma(ASPS) and transcription factor enhancer 3 positive; both cases were myogenic tumors. Literature review (including this paper) showed that there were in total 14 cases with DMD combined with myogenic tumors including 13 cases of rhabdomyosarcoma (RMS) and 1 case of ASPS. All of them are male, and the age of onset of the tumors was 4-17 years. Pathological subtypes were described in 6 cases of ARMS and 5 cases of embryonal RMS, and were not described in 2 cases. The 9 cases described all had large deletions in the DMD gene which can change the reading frame of the DMD gene, and all gene mutations did not exceed exon 62. Conclusions:DMD gene with deletion may increase the risk of having myogenic tumors, and RMS is more common, which is manifested as painless mass in early stage. All DMD gene deletions do not exceed exon 62 and lead to change of the gene reading frame with severe clinical phenotype and degenerative changes in muscle function.

Cryopreservation is the primary technique for in vitro preservation of allogeneic tissue. However, its success is often hindered by factors such as low temperature, ischemia, and hypoxia. This study investigated the potential of Sini Decoction, known for its antioxidant and anti-apoptotic properties, to reduce cryopreservation-induced injury in rats' sciatic nerves. Sini Decoction was prepared according to the Chinese Pharmacopoeia, and its cytotoxicity on Rsc96 cells was assessed by using the CCK-8 method. Sini Decoction at concentrations of 4, 8, and 16 mg·mL~(-1), termed as low-(SL), medium-(SM), and high-(SH) doses group, was used for cryopreservation of rats' sciatic nerves. A normal control(NC) group and a fresh nerve control(fresh) group were set. Flow cytometry and TUNEL staining were used to detect the apoptosis of neural tissue cells after cryopreservation. Western blot was used to detect the expression of apoptosis-related proteins(Bcl-2, Bax, caspase-3, and caspase-8) and nerve regeneration proteins(NGF and BDNF) in vitro after cryopreservation. Oxidative damage of neural tissue after cryopreservation was evaluated by measuring levels of GSH, SOD, MDA, ROS, and ATP. Cryopreserved nerves were then used for allogeneic transplantation. One week after transplantation, CD4~+ and CD8~+ fluorescent double staining assessed inflammatory cell invasion in the transplanted nerve segment, and ELISA evaluated the expression of serum inflammatory factors(IL-1, IFN-γ, and TNF-α) in recipients. Twenty weeks after transplantation, electrophysiology and NF200 neurofilament staining were used to evaluate nerve regeneration. RESULTS:: showed that Sini Decoction at concentrations of below 32 mg·mL~(-1) exhibited no cytotoxicity to Rsc96 cells. During in vitro nerve cryopreservation, Sini Decoction significantly reduced cell apoptosis, ROS, and MDA production compared to the NC group. In the SH group, the protein expression of NGF and BDNF in vitro, as well as ATP, SOD, and GSH production, were significantly increased. In the rejection reaction one week after transplantation, compared to the fresh nerve transplantation group, the SL and SM groups showed reduced CD4~+ and CD8~+ T cell invasion in the transplanted nerve segment and down-regulated IL-1, IFN-γ, and TNF-α expression in recipient serum. Twenty weeks after transplantation, the electrophysiological test results of CMAP, NCV, and NF200 neurofilament protein fluorescent staining in the SM and SH groups were superior to those in the NC and fresh groups. These findings indicate that Sini Decoction offers protective benefits in the cryopreservation of rats' sciatic nerves and holds significant potential for the in vitro preservation of tissue and organs.
Animals ; Apoptosis/drug effects* ; Rats ; Oxidative Stress/drug effects* ; Sciatic Nerve/cytology* ; Cryopreservation ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Rats, Sprague-Dawley ; Protective Agents/pharmacology*

The chemical constituents were systematically separated from the roots of Berberis polyantha by various chromatographic methods, including silica gel column chromatography, HP20 column chromatography, polyamide column chromatography, reversed-phase C_(18) column chromatography, and preparative high-performance liquid chromatography. The structures of the compounds were identified by physicochemical properties and spectroscopic techniques(1D NMR, 2D NMR, UV, MS, and CD). Four phenylpropanoids were isolated from the methanol extract of the roots of B. polyantha, and they were identified as(2R)-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone-O-β-D-glucopyranoside(1), methyl 4-hydroxy-3,5-dimethoxybenzoate(2),(+)-syringaresinol(3), and syringaresinol-4-O-β-D-glucopyranoside(4). Compound 1 was a new compound, and other compounds were isolated from this plant for the first time. The anti-inflammatory activity of these compounds was evaluated based on the release of nitric oxide(NO) in the culture of lipopolysaccharide(LPS)-induced RAW264.7 macrophages. At a concentration of 10 μmol·L~(-1), all the four compounds inhibited the LPS-induced release of NO in RAW264.7 cells, demonstrating potential anti-inflammatory properties.
Plant Roots/chemistry* ; Animals ; Mice ; Berberis/chemistry* ; RAW 264.7 Cells ; Macrophages/immunology* ; Drugs, Chinese Herbal/isolation & purification* ; Nitric Oxide/metabolism* ; Molecular Structure ; Anti-Inflammatory Agents/isolation & purification*

This study aims to explore the pharmacodynamic material basis of Jinbei Oral Liquid(JBOL) against idiopathic pulmonary fibrosis(IPF) based on serum pharmacochemistry and network pharmacology. The ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q-TOF-MS/MS) technology was employed to analyze and identify the components absorbed into rat blood after oral administration of JBOL. Combined with network pharmacology, the study explored the pharmacodynamic material basis and potential mechanism of JBOL against IPF through protein-protein interaction(PPI) network construction, "component-target-pathway" analysis, Gene Ontology(GO) functional enrichment, and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis. First, a total of 114 compounds were rapidly identified in JBOL extract according to the exact relative molecular mass, fragment ions, and other information of the compounds with the use of reference substances and a self-built compound database. Second, on this basis, 70 prototype components in blood were recognized by comparing blank serum with drug-containing serum samples, including 28 flavonoids, 25 organic acids, 4 saponins, 4 alkaloids, and 9 others. Finally, using these components absorbed into blood as candidates, the study obtained 212 potential targets of JBOL against IPF. The anti-IPF mechanism might involve the action of active ingredients such as glycyrrhetinic acid, cryptotanshinone, salvianolic acid B, and forsythoside A on core targets like AKT1, TNF, and ALB and thereby the regulation of multiple signaling pathways including PI3K/AKT, HIF-1, and TNF. In conclusion, JBOL exerts the anti-IPF effect through multiple components, targets, and pathways. The results would provide a reference for further study on pharmacodynamic material basis and pharmacological mechanism of JBOL.
Drugs, Chinese Herbal/pharmacokinetics* ; Animals ; Tandem Mass Spectrometry ; Network Pharmacology ; Rats ; Chromatography, High Pressure Liquid ; Rats, Sprague-Dawley ; Male ; Idiopathic Pulmonary Fibrosis/metabolism* ; Humans ; Administration, Oral ; Protein Interaction Maps/drug effects* ; Signal Transduction/drug effects*