Objective:To investigate the role of platelet-derived zyxin in promoting tumor migration by platelets.Methods:The gene expression profile of platelets was analyzed from cancer patients by using the GEO database.Isolated platelets from wild-type(WT)and Zyx-/-mice were co-cultured with B16F10 cells labeled with green fluorescence to investigate the influence of zyxin deficiency on tumor cell migration,invasion,and wound healing.Optical microscopy was employed to evaluate the impact of zyxin deficiency on epithelial-mesenchymal transition(EMT)in B16F10 cells induced by platelets.Employing specific markers to label platelets,fluorescence confocal microscopy was utilized to investigate the impact of platelet-derived zyxin on the binding between tumor cells and platelets.And an aggregometer was employed to observe the influence of zyxin deficiency on tumor cell-induced platelet aggregation.Results:Compared to platelets from healthy volunteers,zyxin was upregulated in platelets from cancer patients.Zyx-/-mouse platelets exhibited a significant reduction in tumor cell invasion and migration,impaired wound healing,and delayed tumor cell EMT compared to WT mouse platelets.Additionally,zyxin deficiency attenuated the interaction between platelets and tumor cells,and diminished the capacity for tumor cell-induced platelet aggregation.Conclusion:Platelet-derived zyxin deficiency diminishes platelet-tumor cell interactions and weakens the ability of tumor cell-induced platelet aggregation,ultimately suppressing tumor cell migration.

A recombinant adenovirus(rAd)for expression of Mycobacterium tuberculosis(M.tb)c-di-AMP phosphodiesterase CnpB was constructed,and its induced humoral immune response was detected.The codon-optimized gene of M.tb CnpB was cloned into the adenoviral plasmid pcADV.The recombinant plasmid pcADV-CnpB was transfected into HEK293T cells,and expression was detected with Western blot.The recombinant plasmid pcADV-CnpB and the backbone plasmid were co-transfected into HEK293T cells to obtain the recombinant adenovirus rAd-CnpB.rAd-CnpB was amplified in HEK293T cells,and the target protein expression of rAd-CnpB was detected with Western blot and immunofluorescence.Mice were immunized with rAd-CnpB intranasally,and their sera and bronchoalveolar lavage fluid(BALF)were collected.ELISA was used to detect levels of antigen-specific antibodies.Restriction enzyme digestion and sequencing indicated that the recombinant plasmid pcADV-CnpB was successfully constructed and led to protein expression in eukaryotic cells.rAd-CnpB was packaged and produced in HEK293T cells.After amplification and purification,rAd-CnpB with a titer of 5.53×1010 PFU/mL was obtained.rAd-CnpB led to CnpB expression in HEK293T cells.Intranasal immunization with rAd-CnpB increased levels of IgG and secretory IgA in BALF and led to high levels of IgG in sera.rAd-CnpB,the recombinant adenovirus for expression of c-di-AMP phosphodiesterase CnpB was successfully constructed,and was found to induce antigen-specific humoral and mucosal immune responses through mucosal immunization.Thus,rAd-CnpB may be used in further research on new TB vaccine strategies.

This study assessed the role and mechanism of the recombinant Bacillus Calmette-Gue?rin vaccine(rBCG)with c-di-AMP adjuvant in regulating metabolism and immunity in epididymal white adipose(eWAT)in mice.Male C57BL/6 mice were intravenously immunized with BCG and rBCG,and their body weights were monitored.eWAT was isolated from the mice,and the stromal vascular fractions(SVFs)cell number was counted with a hemocytometer.Sections of mouse adipose tissue were prepared,and the size,number,and morphology of eWAT adipocytes and crown-like structure(CLS)formation were compared under a microscope after HE staining.The transcription levels of lipid metabolism-associated factors,cytokines and aging-associated genes in each group were determined with qRT-PCR.The body weights of mice gradually increased after immunization with BCG and rBCG.The proportions of eWAT increased,and the SVFs cell number decreased,in rBCG immunized mice.HE staining indicated that BCG immunization promoted hyperplasia,whereas rBCG immunization promoted hypertrophy of eWAT adipocytes;moreover,both BCG and rBCG immunization induced CLS formation in eWAT.The qRT-PCR results indicated that rBCG immunization inhibited the expression of genes associated with lipolysis and energy expenditure in eWAT.BCG immunization had little effect on cytokine transcription,whereas rBCG significantly induced the transcription of IFN-γ and IL-1Ra,and inhibited that of IL-15 and IL-2,but did not induce the expression of aging-associated genes.Thus,rBCG immunization induced eWAT adipocyte hypertrophy,which was associated with the inhibition of eWAT lipolysis and the regulation of cytokine expression.

Background and Aims:Autoimmune thyroid disease(AITD)are closely associated with metabolic dysregulation,but the causal role of specific metabolites remains unclear.This study aimed to systematically evaluate the causal relationships between approximately 1 400 blood metabolites and two major AITD subtypes-Graves'disease(GD)and Hashimoto's thyroiditis(HT)-using a two-sample Mendelian randomization(MR)approach,to identify potential risk or protective metabolites and provide genetic evidence for mechanistic studies and targeted metabolic interventions.Methods:Summary-level genome-wide association study(GWAS)data for blood metabolites and AITDs were analyzed using inverse-variance weighted MR as the primary method,supplemented by MR-Egger,weighted median,and mode-based methods.Heterogeneity,pleiotropy,and robustness were assessed through Cochran's Q test,horizontal pleiotropy test,and leave-one-out analyses.Results:Forty-nine metabolites showed significant causal associations with GD and 89 with HT.Hexanoylglutamine and ceramide(d18∶1/16∶0)were identified as GD risk factors,while N2,N2-dimethylguanosine and β-hydroxyisovalerylcarnitine were protective.Pregnanediol sulfate and theobromine were associated with increased HT risk,whereas dihomo-linolenate(20:3n3 or n6)and caprylate appeared protective.The α-ketoglutarate/succinate ratio was positively associated with both diseases,suggesting a shared metabolic risk pathway.Conclusion:This MR study provides genetic evidence supporting causal links between multiple blood metabolites and GD or HT.Several metabolites may serve as predictive or protective biomarkers,offering novel insights into the pathophysiology,early screening,and personalized metabolic intervention strategies for AITDs.

Objective:To investigate the relationship between the severity of pharyngolaryngeal sensory impairment and swallowing biomechanics as well as the risk of penetration-aspiration in patients with dysphagia following infratentorial stroke.Methods:This retrospective cross-sectional study enrolled 51 patients with dysphagia following infratentorial stroke hospitalized in the Department of Rehabilitation Medicine of The Third Affiliated Hospital of Sun Yat-sen University between January 2022 and December 2023. Participants were categorized into three groups: normal sensation group [15 males, 2 females; age range 29-76 (56.0±13.3)years], diminished sensation group[16 males, 3 females; age range 38-80(62.0±11.8)years], and absent sensation group [14 males, 1 female; age range 44-75 (60.0±9.7)years]. All patients underwent laryngoscopy and videofluoroscopic swallowing study, which included pharyngolaryngeal sensory testing and Penetration-Aspiration Scale assessment. Swallowing temporal parameters were quantitatively analyzed. Group comparisons for different variable types were conducted using the Chi-square test, one-way ANOVA, and the Kruskal-Wallis test. The correlation between sensory groups and Penetration-Aspiration Scale scores was assessed using Spearman′s correlation analysis. Logistic regression was employed to analyze the impact of pharyngolaryngeal sensory function on penetration-aspiration events.Results:Among the 51 patients, 33.33% (17/51) had normal pharyngolaryngeal sensation, while, 66.67% (34/51) exhibited sensory impairment. The normal sensation group exhibited a significantly longer laryngeal vestibule closure (LVC) time [792 (643, 1 205) ms] compared to the diminished [528 (380, 776) ms] and absent sensation groups [380 (322, 404) ms] ( H=6.502, P=0.039). Additionally, the upper esophageal sphincter opening time was longer in the normal sensation group than in the absent sensation group [528 (371, 710) ms vs 182 (0, 710) ms, H=6.003, P=0.049]. Correlation analysis indicated a significant negative correlation between the severity of sensory impairment and Penetration-Aspiration Scale scores ( r=-0.366, P=0.008). Logistic regression analysis demonstrated that greater sensory impairment was an independent risk factor for penetration-aspiration ( OR=9.29, 95%CI=1.57-54.77, P=0.014). Conclusion:Pharyngolaryngeal sensory deficits are common after infratentorial stroke dysphagia and are significantly associated with impaired swallowing biomechanics and increased aspiration risk. The severity of sensory deficit is a key determinant of penetration-aspiration risk, highlighting its value in risk stratification and therapeutic decision-making for dysphagia.

Based on the results of the National Drug Sampling and Inspection Programme,we summarized the history,the standard collection,the production enterprises and the dosage form specifications,the quality standard study,the pharmacological and pharmacodynamic study,and the clinical application study of Wuwei Qingzhuo preparation of Mongolian medicine and Shiliu Jianwei preparation of Zang medicine,to provide the basis for improved quality standards for both preparations.The development of these two preparations was searched and analyzed through literature.The available information shows that there is very little research on the two preparations and insufficient pharmacological experimental and clinical experimental data.The two preparations are basically the same in prescription and efficacy.However,the quality standards are very different,which are not conducive to the quality control of the two and their related dosage forms.And it is suggested that the Chinese Pharmacopoeia should take the situation of this category into comprehensive consideration,and unify the quality standards of the two preparations.

With the prevalence of kidney disease continues to rise,it has become a serious global public health issue that significantly impacts patients'quality of life.The complex pathogenesis of kidney disea-ses presents challenges for its prevention and treatment.Research has shown that imbalances in metabolic homeostasis often affect kidney function,leading to renal damage.Peroxisome proliferator-activated re-ceptors(PPARs),as transcription factors activated by endogenous ligands,play a crucial role in regula-ting metabolic molecular networks,particularly in oxidative phosphorylation,lipid metabolism,and glu-cose metabolism.Activating PPARs can improve mitochondrial damage,promote tatty acid breakdown,and alleviate insulin resistance,thereby restoring renal metabolic function and mitigating kidney damage.Although various small-molecule drugs that activate PPARs have been developed,adverse reactions have been observed during clinical trials.Currently,there is still a lack of safe and effective treatment options for kidney diseases.A detailed analysis of the molecular mechanisms by which the PPAR family regulates renal cell energy metabolism,as well as the search for and development of new small-molecule drugs tar-geting PPARs and their regulated downstream metabolic pathways,is of significant importance for under-standing and treating kidney diseases.On the other hand,clarifying the metabolic changes during the progression of different kidney diseases and applying targeted PPAR-based drugs or strategies for their treatment is particularly important.Here we summarize how PPAR family members regulate target gene transcription and their roles in the remodeling of oxidative phosphorylation and lipid/glucose metabolism,as well as the impact of changes in PPAR expression or activity on acute and chronic kidney diseases and age-related kidney conditions.This knowledge may provide new insights and theoretical support for the prevention and treatment of kidney diseases.

Objective:To investigate the role of platelet-derived zyxin in promoting tumor migration by platelets.Methods:The gene expression profile of platelets was analyzed from cancer patients by using the GEO database.Isolated platelets from wild-type(WT)and Zyx-/-mice were co-cultured with B16F10 cells labeled with green fluorescence to investigate the influence of zyxin deficiency on tumor cell migration,invasion,and wound healing.Optical microscopy was employed to evaluate the impact of zyxin deficiency on epithelial-mesenchymal transition(EMT)in B16F10 cells induced by platelets.Employing specific markers to label platelets,fluorescence confocal microscopy was utilized to investigate the impact of platelet-derived zyxin on the binding between tumor cells and platelets.And an aggregometer was employed to observe the influence of zyxin deficiency on tumor cell-induced platelet aggregation.Results:Compared to platelets from healthy volunteers,zyxin was upregulated in platelets from cancer patients.Zyx-/-mouse platelets exhibited a significant reduction in tumor cell invasion and migration,impaired wound healing,and delayed tumor cell EMT compared to WT mouse platelets.Additionally,zyxin deficiency attenuated the interaction between platelets and tumor cells,and diminished the capacity for tumor cell-induced platelet aggregation.Conclusion:Platelet-derived zyxin deficiency diminishes platelet-tumor cell interactions and weakens the ability of tumor cell-induced platelet aggregation,ultimately suppressing tumor cell migration.

Objective:To analyze the pathogenicity and clinical phenotype associated with a newly identified heterozygous variant in the ZEB1 gene that causes posterior pleomorphic corneal dystrophy (PPCD). Methods:A pedigree study was conducted.Clinical data of four people in 2 generations from one family with PPCD who visited Henan Eye Hospital in October 2023 were collected, including 3 patients. Relevant ophthalmic examinations were performed.Best corrected visual acuity, slit lamp microscopy, intraocular pressure, Pentacam corneal topography, Corvis ST corneal biomechanics analyzer, corneal endothelial microscopy, swept-source anterior segment coherence optical tomography (CASIA), laser scanning confocal microscopy, and ultra-wide-field fundus photography were performed to examine clinical phenotypes.Peripheral venous blood samples were collected from family members to extract genomic DNA, and whole exome sequencing was performed.Sanger sequencing and pedigree co-segregation analysis were carried out.Conservation analysis was performed using GERP+ + and Clustal Omega software, and the pathogenicity of the variant was assessed according to American College of Medical Genetics and Genomics (ACMG) guidelines.This study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[15]).All subjects or guardian signed informed consent.Results:This family conformed to autosomal dominant inheritance.Under a slit-lamp microscope, corneal endothelial vesicular lesions in both eyes could be seen in the proband, her father and her brother.Under a laser scanning confocal microscope, endothelial cells were missing at the lesions, and some were crater-like changes, and some lesions were circular or elliptical vesicular, and no other systemic abnormalities were observed.The ocular and physical examination of the proband's mother showed no abnormalities.Genetic testing results showed that the proband, her father and her brother all carried the ZEB1c.790G>A (p.Gly264Arg) heterozygous variant, but her mother did not carry the variantion.Sanger sequencing verified that this variantion was co-segregated within the family.The variantion is a newly discovered missense mutation that had not been reported in the Thousand Genomes Project, Genome Aggregation Database, and ExAC database.The prediction results of the variant by MutationTaster, SIFT, PROVEAN, VESST3, DANN, FATHMM-MKL, CADD, fitCons and other software were harmful, and GERP+ +, Weblogo, Clustal Omega analysis showed that the amino acids affected by the variant were highly conservative.According to the ACMG Guidelines, this variation was possible pathogenic.Conclusions:The identification of the missense mutation c. 790G>A (p.Gly264Arg) in the ZEB1 gene within this PPCD family provides new insights into the genetic basis of PPCD and the variant may be the pathogenic variant of in this family.

Background and Aims:Autoimmune thyroid disease(AITD)are closely associated with metabolic dysregulation,but the causal role of specific metabolites remains unclear.This study aimed to systematically evaluate the causal relationships between approximately 1 400 blood metabolites and two major AITD subtypes-Graves'disease(GD)and Hashimoto's thyroiditis(HT)-using a two-sample Mendelian randomization(MR)approach,to identify potential risk or protective metabolites and provide genetic evidence for mechanistic studies and targeted metabolic interventions.Methods:Summary-level genome-wide association study(GWAS)data for blood metabolites and AITDs were analyzed using inverse-variance weighted MR as the primary method,supplemented by MR-Egger,weighted median,and mode-based methods.Heterogeneity,pleiotropy,and robustness were assessed through Cochran's Q test,horizontal pleiotropy test,and leave-one-out analyses.Results:Forty-nine metabolites showed significant causal associations with GD and 89 with HT.Hexanoylglutamine and ceramide(d18∶1/16∶0)were identified as GD risk factors,while N2,N2-dimethylguanosine and β-hydroxyisovalerylcarnitine were protective.Pregnanediol sulfate and theobromine were associated with increased HT risk,whereas dihomo-linolenate(20:3n3 or n6)and caprylate appeared protective.The α-ketoglutarate/succinate ratio was positively associated with both diseases,suggesting a shared metabolic risk pathway.Conclusion:This MR study provides genetic evidence supporting causal links between multiple blood metabolites and GD or HT.Several metabolites may serve as predictive or protective biomarkers,offering novel insights into the pathophysiology,early screening,and personalized metabolic intervention strategies for AITDs.