Aim To perform high-throughput screen-ing of immunomodulatory traditional Chinese medicine(TCMs)based on an in vitro B cell differentiation-antibody secretion model,identifying active herbal candidates with immune-enhancing properties to pro-vide novel therapeutic options and theoretical support for influenza virus treatment in immunocompromised in-dividuals.Methods B cells were stimulated with dif-ferent concentrations of cytosine-phosphate-guanine oli-godeoxynucleotide 2006(CpG)and nterleukin-2(IL-2)to promote proliferation,differentiation,and anti-body secretion,and the effects of varying concentra-tions of the solvent DMSO were also evaluated.The op-timal conditions for the B cell differentiation-anti-body secretion model were determined based on the se-cretion levels of three antibody isotypes.The feasibility of the model was further validated using rapamycin,a known B cell function inhibitor.On this basis,a high-throughput screening platform for immunomodulatory a-gents was optimized and established.Subsequently,the immune-enhancing activity of 465 polarity extract from TCMs was evaluated.Results The optimal con-ditions for the model were determined as 2 mg·L-1 CpG,1.67 × 106 nkat·L-1 IL-2,and DMSO with a volume fraction of 0.1％.Rapamycin effectively inhib-ited B cell differentiation into plasmablast and signifi-cantly reduced antibody production,indicating the reli-ability of the model.Multiple rounds of screening re-vealed that the dichloromethane extract of licorice,the dichloromethane extract of Vinegar-processed Curcumae Rhizoma,the cyclohexane extract of Honey-prepared Radix Asteris,and the aqueous extract of Siphonostegia chinensis Benth were identified to significantly promote both B cell proliferation and differentiation and anti-body secretion at a concentration of 600 μg·L-1.Conclusion This study successfully optimizes an in vitro B cell differentiation-antibody secretion model and identifies several TCM extracts,including licorice,with potential immune-enhancing activity.

Progressive symmetric erythrokeratodermia(PSEK)is a rare hereditary skin disease charac-terized by symmetrical erythema,hyperkeratosis and multiorgan lesions.Its clinical phenotypes are highly heterogeneous and may be accompanied by symptoms such as thrombocytopenia,which can be fatal in se-vere cases.The genotype-phenotype association mechanism of PSEK is extremely complex.Currently,it is known that mutations in multiple genes such as GJB3,KDSR,and KRT83 can cause the disease.A-mong them,3-ketodihydrosphingosine reductase(KDSR)has been found to harbor nearly 20 clinical mu-tations.These mutations interfere with the de novo ceramide synthesis pathway,disrupt the homeostasis of the skin barrier,and cause platelet production disorders and multi-organ lesions,making it a current research hotspot in the molecular mechanism of PSEK.The pathogenic mutations of KDSR are widely and uniformly distributed throughout the entire protein,rather than being limited to the traditionally recog-nized active center,suggesting that the impairment of the KDSR enzymatic activity is not the only cause of PSEK.In view of this,this study selected four typical mutants of KDSR(KDSRQG55-56R,KDSRn38C,KDSRY186F,KDSRG182S),and first used recombinant expression technology to prepare pure and homoge-neous mutant proteins.Subsequently,thermal stability experiments as well as oligomerization analysis were conducted on these four mutant proteins.The results showed that the Tm values of the four mutants were significantly lower than that of the wild type.Particularly,KDSRF138C and KDSRQG55-56R were nearly completely denatured at physiological temperature.This result was perfectly consistent with the further Rosetta energy analysis.In conclusion,this study took several pathological mutations of the PSEK patho-genic factor KDSR as the research object and discovered that the conformational stability of KDSR might be closely related to the occurrence of PSEK pathogenicity,indicating that the imbalance of conformation-al homeostasis is very likely to be one of the common contributing factors of many genetic diseases,inclu-ding PSEK.This provides a new theoretical basis and reference for explaining the molecular mechanism of genotype-phenotype heterogeneity in many genetic diseases.

Aim To perform high-throughput screen-ing of immunomodulatory traditional Chinese medicine(TCMs)based on an in vitro B cell differentiation-antibody secretion model,identifying active herbal candidates with immune-enhancing properties to pro-vide novel therapeutic options and theoretical support for influenza virus treatment in immunocompromised in-dividuals.Methods B cells were stimulated with dif-ferent concentrations of cytosine-phosphate-guanine oli-godeoxynucleotide 2006(CpG)and nterleukin-2(IL-2)to promote proliferation,differentiation,and anti-body secretion,and the effects of varying concentra-tions of the solvent DMSO were also evaluated.The op-timal conditions for the B cell differentiation-anti-body secretion model were determined based on the se-cretion levels of three antibody isotypes.The feasibility of the model was further validated using rapamycin,a known B cell function inhibitor.On this basis,a high-throughput screening platform for immunomodulatory a-gents was optimized and established.Subsequently,the immune-enhancing activity of 465 polarity extract from TCMs was evaluated.Results The optimal con-ditions for the model were determined as 2 mg·L-1 CpG,1.67 × 106 nkat·L-1 IL-2,and DMSO with a volume fraction of 0.1％.Rapamycin effectively inhib-ited B cell differentiation into plasmablast and signifi-cantly reduced antibody production,indicating the reli-ability of the model.Multiple rounds of screening re-vealed that the dichloromethane extract of licorice,the dichloromethane extract of Vinegar-processed Curcumae Rhizoma,the cyclohexane extract of Honey-prepared Radix Asteris,and the aqueous extract of Siphonostegia chinensis Benth were identified to significantly promote both B cell proliferation and differentiation and anti-body secretion at a concentration of 600 μg·L-1.Conclusion This study successfully optimizes an in vitro B cell differentiation-antibody secretion model and identifies several TCM extracts,including licorice,with potential immune-enhancing activity.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

Objective:To construct an integrated management model for early screening and diagnosis of PCa based on the In-novative Care for Chronic Conditions Framework(ICCC)and the 1+1 contract-based tiered diagnosis and treatment system(TDTS)in China.Methods:Based on the 1+1 contract-based TDTS platform,we conducted PCa screening for the male residents aged 60 years and above during health check-ups in Pujin Community Health Center from January 1,2023 to December 31,2023.For those with abnormal total prostate-specific antigen(tPSA)≥4 μg/L,we promptly referred them to higher-level hospitals for further diagno-sis and treatment via the two-way referral green channel platform and information sharing service using the 1+1 contract model.We further analyzed the relevant data on screening and diagnosis.Results:A total of 4 080 males aged 71.39±5.059 years received PCa screening from January to December 2023.PSA screening was performed in 43.96％of the male residents,revealing 654 cases of PSA abnormality,with a PSA positivity rate of 16.03％,which was higher than that found in the previous large-scale PCa screenings in other regions of China.Among the males with PSA abnormality,292(44.65％)expressed their willingness for medical referral,while the others did not seek further medical attention for reasons of being asymptomatic,low awareness of the disease,no accompany for medical visits,and concerns about further costs of diagnosis and treatment.Prostate biopsy was recommended to 154 cases after further examinations,which was accepted by 92(59.74％).Fifty-eight cases were diagnosed with Pa,and thedetection rate reached 63.04％.Conclusion:The integrated management model for PSA examination-based early screening and diagnosis of PCa using the 1+1 contract-based TDTS platform is plays a significant role in enhancing peoples awareness and knowledge of PCa and improving the early detection rate of the malignancy.

Progressive symmetric erythrokeratodermia(PSEK)is a rare hereditary skin disease charac-terized by symmetrical erythema,hyperkeratosis and multiorgan lesions.Its clinical phenotypes are highly heterogeneous and may be accompanied by symptoms such as thrombocytopenia,which can be fatal in se-vere cases.The genotype-phenotype association mechanism of PSEK is extremely complex.Currently,it is known that mutations in multiple genes such as GJB3,KDSR,and KRT83 can cause the disease.A-mong them,3-ketodihydrosphingosine reductase(KDSR)has been found to harbor nearly 20 clinical mu-tations.These mutations interfere with the de novo ceramide synthesis pathway,disrupt the homeostasis of the skin barrier,and cause platelet production disorders and multi-organ lesions,making it a current research hotspot in the molecular mechanism of PSEK.The pathogenic mutations of KDSR are widely and uniformly distributed throughout the entire protein,rather than being limited to the traditionally recog-nized active center,suggesting that the impairment of the KDSR enzymatic activity is not the only cause of PSEK.In view of this,this study selected four typical mutants of KDSR(KDSRQG55-56R,KDSRn38C,KDSRY186F,KDSRG182S),and first used recombinant expression technology to prepare pure and homoge-neous mutant proteins.Subsequently,thermal stability experiments as well as oligomerization analysis were conducted on these four mutant proteins.The results showed that the Tm values of the four mutants were significantly lower than that of the wild type.Particularly,KDSRF138C and KDSRQG55-56R were nearly completely denatured at physiological temperature.This result was perfectly consistent with the further Rosetta energy analysis.In conclusion,this study took several pathological mutations of the PSEK patho-genic factor KDSR as the research object and discovered that the conformational stability of KDSR might be closely related to the occurrence of PSEK pathogenicity,indicating that the imbalance of conformation-al homeostasis is very likely to be one of the common contributing factors of many genetic diseases,inclu-ding PSEK.This provides a new theoretical basis and reference for explaining the molecular mechanism of genotype-phenotype heterogeneity in many genetic diseases.

Objective:To explore the significance of thrombus biomarkers in evaluating the progression of immunoglobulin A vasculitis(IgAV)in children.Methods:A total of 193 children who were diagnosed as IgAV from September 2021 to June 2023 in the Children's Hospital of Soochow University were enrolled.The levels of plasma thrombomodulin(TM),thrombin-antithrombin complex(TAT),plasmin-α 2-plasmin inhibitor complex(PIC),tissue plasminogen activator-plasminogen activator inhibitor-1 complex(t-PAIC)and D-dimer(D-D)were analyzed retrospectively.And,140 healthy children were selected as controls during the same period.The receiver operating characteristic(ROC)curves were drawn to analyze the role of thrombus parameters in estimating the progression of IgAV in children.Univariate and multivariate logistic regression analysis were used to assess the independent risk factors influencing the progression of pediatric IgAV in acute phase.Results:The levels of D-D,TAT,PIC and t-PAIC in plasma of IgAV group were higher than those in control group(all P＜0.001).The levels of D-D,TAT and PIC in acute phase children were significantly higher than those in non acute phase children(all P＜0.001),while the levels of kidney injury related indicators such as 24h-UTP,urine albumin/creatinine ratio,positive urinary blood on dipstick,serum creatinine and cystatin C were lower(all P＜0.05).ROC analysis showed that the area under curve(AUC)of PIC was 0.743 when the cut-off value was 0.93 μg/ml with 71.8％sensitivity and 78.3％specificity,while the AUC of D-D was 0.756 when the cut-off value was 550.0 μg/L with 81.3％sensitivity and 73.4％specificity.Univariate and multivariate logistic regression analysis showed that PIC ≥ 0.93 μg/ml(OR=4.64,P=0.012)and D-D ≥ 550.0 μg/L(OR=3.60,P=0.035)were the independent risk factors for the progression of IgAV in acute phase.Conclusion:The pediatric patients with IgAV have shown hyperfibrinolysis in the acute stage.Furthermore,the levels of PIC and D-D should be of diagnostic value for evaluating the progression of IgAV in the acute phase.

The genome tagging project (GTP) plays a pivotal role in addressing a critical gap in the understanding of protein functions. Within this framework, we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411 (HA-tagged Ssp411) mouse model. This model is instrumental in probing the expression and function of Ssp411. Our research revealed that Ssp411 is expressed in the round spermatids, elongating spermatids, elongated spermatids, and epididymal spermatozoa. The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis. Nevertheless, rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions. Ssp411 is not detectable in metaphase II (MII) oocytes, zygotes, or 2-cell stage embryos, highlighting its intricate role in early embryonic development. These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP, fostering groundbreaking advancements in the fields of spermiogenesis and reproductive biology.
Animals ; Female ; Humans ; Male ; Mice ; Spermatids/metabolism* ; Spermatogenesis/physiology* ; Spermatozoa/metabolism* ; Thioredoxins/genetics*

To evaluate the therapeutic effect of a modified Devine procedure with a subcutaneous sliding fixation method for the treatment of congenital concealed penis, we retrospectively selected 45 patients with congenital concealed penises who were admitted to General Hospital of Ningxia Medical University (Yinchuan, China) between September 2020 and November 2023. In all cases, the penis was observed to be short, and retracting the skin at the base revealed a normal penile body, which immediately returned to its original position upon release. All patients underwent the modified Devine procedure with subcutaneous sliding fixation and completed a 12-week postoperative follow-up. A statistically significant increase in penile length was observed postoperatively, with the median length increasing from 4.0 (interquartile range [IQR]: 3.5-4.8; 95% confidence interval [CI]: 3.9-4.4) cm to 8.0 (IQR: 7.8-8.0; 95% CI: 7.7-7.9) cm, with P < 0.001. The parents were satisfied with the outcomes, including increased penile length, improved hygiene, and enhanced esthetics. Except for mild foreskin edema in all cases, no complications (such as infections, skin necrosis, or penile retraction) were observed. The edema was resolved within 4 weeks after the operation. This study demonstrates that the modified Devine procedure utilizing the subcutaneous sliding fixation method yields excellent outcomes with minimal postoperative complications, reduced penile retraction, and high satisfaction rates among patients and their families.
Humans ; Male ; Penis/abnormalities* ; Retrospective Studies ; Urologic Surgical Procedures, Male/methods* ; Treatment Outcome ; Child ; Plastic Surgery Procedures/methods*

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*