ObjectiveThe accuracy of Y-chromosome haplogroup assignment is crucial for tracing paternal lineage in male samples. With the advancement of high-throughput sequencing technologies, high-density Y-SNP genotyping from whole-genome or array-based data has become a standard method for determiningY-chromosome haplogroups. This study systematically evaluated the performance of 4 commonly used high-density SNP genotyping systems—namely, the Global Screening Array (GSA), Chinese Genotyping Array (CGA), Affymetrix array, and the 1240K capture panel—for haplogroup assignment. This work provides a reference for data comparison across different systems. MethodsWe extracted genotype data for the 4 Y-SNP panels from 30× whole-genome sequencing (WGS) data of 1 590 male samples from the 1000 Genomes Project. Additionally, GSA array genotype data from 384 relative pairs (spanning 1st- to 12th-degree relationships) from 109 Chinese Han families were collected. Haplogroup assignment was performed using Y-LineageTracker v1.3.0 software. We assessed the concordance and resolution of haplogroup assignments between the four Y-SNP panels and the WGS data. The consistency and resolution of haplogroup assignments were also evaluated for both the 1000 Genomes Project samples and the 109 family samples collected in this study. Furthermore, the impact of varying numbers of Y-SNPs on haplogroup assignment was examined. ResultsThe GSA and CGA panels demonstrated superior resolution and discrimination of haplogroup subclades compared with the other two panels. The haplogroup assignments from the GSA, CGA, and 1240K panels showed high concordance with WGS data, with consistency rates exceeding 88.70%, whereas the Affymetrix platform exhibited a significantly lower consistency rate of 61.89%. Specifically, the GSA and CGA panels consistently demonstrated superior performance compared with the other two panels in the assignment of haplogroups O-M175 and H-L901, achieving complete concordance (100%) for both haplogroups. In contrast, the Affymetrix panel erroneously assigned all individuals belonging to haplogroup O-M175 to haplogroup K2-M526. Furthermore, its accuracy for haplogroup H-L901 was exceedingly low, at merely 1.41%. This poor performance was characterized by the misassignment of 98.59% of H-L901 samples—specifically, 1.41% to J-M304 and a predominant 97.18% to F-M89. For haplogroup R-M207, all four panels exhibited uniformly high levels of consistency, with concordance values exceeding 94.00%. Notably, for haplogroup E-M96, the 1240K and Affymetrix panels outperformed the GSA and CGA panels in terms of concordance, representing the first instance in which these two panels surpassed the latter. Conversely, for haplogroups J-M304, Q-M242, and I-M170, all 4 panels showed relatively elevated misclassification rates, with the Affymetrix array demonstrating the poorest overall performance. None of the four panels showed any discordant haplogroup assignments among the familial relative pairs analyzed. A positive correlation was observed between the number of Y-SNPs (ranging from 1 000 to 10 000) and classification consistency; however, classification consistency plateaued when the number of Y-SNPs exceeded 10 000. Furthermore, a random sampling analysis conducted on the GSA and CGA panels demonstrated that the haplogroup misclassification rate exhibited negligible fluctuation across the Y-SNP range of 500 to 1 000. Conversely, a marked enhancement in classification consistency was observed as the number of markers increased from 1 000 to 5 000, ultimately reaching a plateau within the interval of 5 000 to 8 000 markers. ConclusionThese findings indicate that the GSA and CGA panels provide high resolution and concordance, delivering reliable Y-haplogroup assignment for forensic investigations.

ObjectiveThe accuracy of Y-chromosome haplogroup assignment is crucial for tracing paternal lineage in male samples. With the advancement of high-throughput sequencing technologies, high-density Y-SNP genotyping from whole-genome or array-based data has become a standard method for determiningY-chromosome haplogroups. This study systematically evaluated the performance of 4 commonly used high-density SNP genotyping systems—namely, the Global Screening Array (GSA), Chinese Genotyping Array (CGA), Affymetrix array, and the 1240K capture panel—for haplogroup assignment. This work provides a reference for data comparison across different systems. MethodsWe extracted genotype data for the 4 Y-SNP panels from 30× whole-genome sequencing (WGS) data of 1 590 male samples from the 1000 Genomes Project. Additionally, GSA array genotype data from 384 relative pairs (spanning 1st- to 12th-degree relationships) from 109 Chinese Han families were collected. Haplogroup assignment was performed using Y-LineageTracker v1.3.0 software. We assessed the concordance and resolution of haplogroup assignments between the four Y-SNP panels and the WGS data. The consistency and resolution of haplogroup assignments were also evaluated for both the 1000 Genomes Project samples and the 109 family samples collected in this study. Furthermore, the impact of varying numbers of Y-SNPs on haplogroup assignment was examined. ResultsThe GSA and CGA panels demonstrated superior resolution and discrimination of haplogroup subclades compared with the other two panels. The haplogroup assignments from the GSA, CGA, and 1240K panels showed high concordance with WGS data, with consistency rates exceeding 88.70%, whereas the Affymetrix platform exhibited a significantly lower consistency rate of 61.89%. Specifically, the GSA and CGA panels consistently demonstrated superior performance compared with the other two panels in the assignment of haplogroups O-M175 and H-L901, achieving complete concordance (100%) for both haplogroups. In contrast, the Affymetrix panel erroneously assigned all individuals belonging to haplogroup O-M175 to haplogroup K2-M526. Furthermore, its accuracy for haplogroup H-L901 was exceedingly low, at merely 1.41%. This poor performance was characterized by the misassignment of 98.59% of H-L901 samples—specifically, 1.41% to J-M304 and a predominant 97.18% to F-M89. For haplogroup R-M207, all four panels exhibited uniformly high levels of consistency, with concordance values exceeding 94.00%. Notably, for haplogroup E-M96, the 1240K and Affymetrix panels outperformed the GSA and CGA panels in terms of concordance, representing the first instance in which these two panels surpassed the latter. Conversely, for haplogroups J-M304, Q-M242, and I-M170, all 4 panels showed relatively elevated misclassification rates, with the Affymetrix array demonstrating the poorest overall performance. None of the four panels showed any discordant haplogroup assignments among the familial relative pairs analyzed. A positive correlation was observed between the number of Y-SNPs (ranging from 1 000 to 10 000) and classification consistency; however, classification consistency plateaued when the number of Y-SNPs exceeded 10 000. Furthermore, a random sampling analysis conducted on the GSA and CGA panels demonstrated that the haplogroup misclassification rate exhibited negligible fluctuation across the Y-SNP range of 500 to 1 000. Conversely, a marked enhancement in classification consistency was observed as the number of markers increased from 1 000 to 5 000, ultimately reaching a plateau within the interval of 5 000 to 8 000 markers. ConclusionThese findings indicate that the GSA and CGA panels provide high resolution and concordance, delivering reliable Y-haplogroup assignment for forensic investigations.

Background: s/Aims: The most common cause of morbidity following pancreatoduodenectomy (PD) is the clinically relevant postoperative pancreatic fistula (CR-POPF). There is currently no universally accepted technique for pancreato-enteric anastomosis. This study aims to compare the Blumgart technique (B-PJ) with the modified Blumgart technique (mB-PJ). Methods: This is a retrospective study involving patients who underwent PD between January 2011 and December 2021. The primary endpoint was to compare the incidence of CR-POPF. Secondary endpoints included major morbidity, length of postoperative stay, rates of reoperation, hospital readmission, postoperative mortality, and predictors of CR-POPF. Propensity score matching (PSM) was employed to minimize potential selection bias. Results: The study included 292 patients. After PSM, the incidence of CR-POPF was not significantly different between B-PJ and mBPJ (18.9% vs. 15.8%, p = 0.566). No statistical differences were observed in the secondary endpoints. Independent predictors of CRPOPF included preoperative cholangitis (odds ratio [OR]: 4.906, 95% confidence interval [CI]: 1.440–16.713, p = 0.011), soft pancreas (OR: 4.259, 95% CI: 1.043–17.376, p = 0.043), and main pancreatic duct size ≤ 3 mm (OR: 5.229, 95% CI: 1.865–14.656, p = 0.002). Conclusions This study did not demonstrate that mB-PJ is superior to B-PJ in reducing the incidence of CR-POPF. Factors such as soft pancreas, main pancreatic duct size, and preoperative cholangitis are identified as independent risk factors for CR-POPF.

Objective By analyzing the gene expression data in TCGA-THCA dataset,the differentially expressed genes related to thyroid carcinoma(THCA)were identified,a risk model was construct,and the role of CCL chemokine family in the development of THCA and its effect on the tumor immune microenvironment were investigated.Methods TCGA-THCA dataset was downloaded and analyzed,including 512 cancer samples and 59 adjacent tissue samples.The differentially expressed genes were obtained by taking the intersection of R packages of differential analysis,and the principal component analysis(PCA),Gene Ontology(GO),and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed.CellMarker,monocle,and CellChat were used to explore the Cell atlas,pseudotemporal trajectory,and cellular communication.Results A total of 698 up-regulated genes and 285 down-regulated genes were identified.PCA showed that there were significant differences in gene expression patterns between the THCA group and the normal group.GO and KEGG enrichment analysis revealed that differentially expressed genes were mainly involved in biological processes such as extracellular matrix tissue,cell junction assembly,and complement system.The results of risk model showed that the CCL genes was significantly correlated with poor prognosis(P=0.005).Single cell analysis revealed that fibroblasts played a key role in the development of THCA,and the expression of CCL gene was significantly up-regulated in fibroblasts(P<0.05).The results of pseudotemporal trajectory analysis showed that CCL gene was differentially expressed during cellular differentiation;the results of cellular communication analysis showed that the interaction between fibroblasts and endothelial cells is crucial in the tumor microenvironment.Conclusion The expression of CCL chemokine family in THCA is closely related to poor prognosis,especially CCL4,which activates the downstream signaling pathways by interacts with CCR receptors of cancer-associated fibroblasts,thereby promoting the development of cancer.These findings provide novel insights for understanding the role of CCL chemokine family in THCA and its regulation of the tumor immune microenvironment.

AIM To prepare fisetin-loaded nanostructured lipid carriers,and to evaluate their in vivo pharmacokinetics.METHODS Ethanol injection method was applied to preparing the nanostructured lipid carriers.With monostearin-phospholipid ratio,monostearin-triacetin ratio and D-α-tocopheryl polyethylene glycol 1000 succinate(TPGS)concentration as influencing factors,encapsulation efficiency as an evaluation index,the formulation was optimized by Box-Behnken response surface method.The crystal form was analyzed by X-ray powder diffraction,after which the morphology was observed by transmission electron microscopy,infrared spectroscopy analysis was performed,the drug relaese was investigated by dialysis bag method,and the stability was determined.Eighteen rats were randomly assigned into 3 groups and given intragastric administration of the 0.5％CMC-Na suspensions of fisetin and its phospholipid complex,nanostructured lipid carriers(150 mg/kg),respectively,after which blood collection was made at0.25,0.5,1,1.5,2,3,4,6,8,12 h,UPLC-MS/MS was adopted in the plasma concentration determination of fisetin,and main pharmacokinetic parameters were calculated.RESULTS The optimal formulation was determined to be 1.56∶1 for monostearin-phospholipid ratio,3.05∶1 for monostearin-triacetin ratio,and 0.2 mg/mL for TPGS concentration.The nearly round fisetin-loaded nanostructured lipid carriers demonstrated the average encapsulation efficiency,drug loading,particle size and Zeta potential of(86.14±1.28)％,(8.96±0.26)％,(212.35±9.04)nm and-(31.13±1.16)mV,respectively.Raw medicine existed in the nanostructured lipid carriers in an amorphous state,preparation process did not affect the hydrogen bonding between raw medicine and phospholipids.The nanostructured lipid carriers displayed the accumulative release rate of 46.12％within 3 h in simulated gastric juice,and that of about 50％within 18 h in simulated intestinal fluid,whose freeze-dried powder exhibited good stability after being placed for 6 months.Compared with raw medicine and phospholipids complex,the nanostructured lipid carriers displayed prolonged tmax,t1/2(P＜0.01)and increased Cmax,AUC0-t,AUC0-∞(P＜0.01),whose relative bioavailability was enhanced to 7.07 times.CONCLUSION Nanostructured lipid carriers can improve the oral bioavailability of fisetin.

Aim To explore the action mechanism of isoquercitrin(IQ)in ameliorating anxiety based on network pharmacology,cellular transcriptomics,molecu-lar docking and animal experiments.Methods The common targets of anxiety disorders and IQ were ob-tained by using relevant databases.The protein-protein interaction network,the biological function and signa-ling pathway enrichment analysis were conducted by u-sing the common targets.Primary hippocampal neurons were cultured in vitro,and corticosterone was added to induce neurons to establish a corticosterone injury mod-el.IQ treatment was added to the culture system,and transcriptomics was used to screen for differentially ex-pressed genes and enrich for differentially expressed pathways.Subsequently,the results were validated by quantitative real-time polymerase chain reaction(qRT-PCR).Possible targets and signaling pathways for IQ treatment on anxiety were speculated and screened u-sing network pharmacology,transcriptomics and molec-ular docking.The anxiety rat model was constructed,and the anxiety state of rats was evaluated after IQ in-tervention,and the protein expression level of hippo-campus was detected to verify the relevant mechanism.Results Network pharmacology,cellular transcrip-tome,and molecular docking analyses revealed that the key mechanism of IQ for anxiety may be related to the BDKRB2/PI3K/Akt signaling pathway.Animal exper-iments showed that IQ was effective in improving anxie-ty behaviour and learning memory ability in rats.IQ increased the movement distance and residence time in the central area of the open field,the time and number percentage of entries into the open arm in the elevated plus maze,and the spontaneous alternations score in the Y maze in rats,and significantly elevated protein expression of BDKRB2,PI3K,Akt and decreased pro-tein expression of NF-κB in the hippocampus.Conclu-sions Isoquercitrin can effectively treat anxiety,and the mechanism of action may be related to the regula-tion of BDKRB2/PI3K/Akt signaling pathway in hip-pocampus.

The scientific research and innovation capabilities of medical students are intrinsically linked to the sustained and high-quality development of national healthcare initiatives.Cultivating outstanding medi-cal students with independent scientific capabilities and innovative consciousness is a critical component in the education and training of high-level medical professionals.Our investigation revealed that within the imperfections of the cultivating model,some faculty and students at medical schools have an insufficient understanding of scientific research and innovation and lack motivation for engaging in such activities,which hinder the progression of scientific research activities.Consequently,we initiated a teaching practice and exploratory study on the"tutorial system"aimed at fostering medical students'scientific research and innovation abilities.Based on the principle of"research informing teaching,teaching and research advan-cing together,"this study implements a"tutorial system"coordinated by tutors,supplemented by graduate and undergraduate student mentors,to cultivate innovative thinking,stimulate interest in scientific re-search,and enhance practical and research skills among medical students.Through collaborative efforts within"scientific research innovation teams,"various educational methods—including preliminary re-search,in-class and extracurricular activities,intra-group and inter-group interactions,and theoretical and practical applications—are employed to improve and strengthen the cultivation of medical students'scientif-ic research and innovation abilities.This study aims to provide valuable references for optimizing medical education management systems and enhancing the quality of medical student training.

Objective:To explore the clinical phenotype and genetic characteristics of a child with hereditary Spastic paraplegia type 52 (SPG52) due to variant of AP4S1 gene. Methods:A child diagnosed with SPG52 at the Department of Pediatrics of the First Affiliated Hospital of Anhui Medical University in May 2010 was selected as the study subject. Whole-exome sequencing (WES) was carried out for the child and his parents. Candidate variants were confirmed by Sanger sequencing. Pathogenicity of the candidate variant was interpreted according to the guidelines from the American College of Medical Genetics and Genomics (ACMG). The study protocol was approved by the Ethics Committee of the Hospital (Ethics No.: PJ2024-04-56).Results:The child had presented with global developmental delay from infancy, and featured progressive lower limb spasticity, contractures, talipes equinovarus, and muscle weakness, but with no significant facial dysmorphism. His first febrile seizure occurred before one year of age, followed by several afebrile seizures. The seizures had remitted after 3 to 4 years of antiepileptic therapy, and electroencephalography was normal. However, he had severe intellectual disability, and MRI revealed reduced white matter. WES identified a homozygous AP4S1 c. 289C>T (p.Arg97*) variant in the child, for which both of his parents were heterozygous carriers. The variant was rated as pathogenic based on the ACMG guidelines. Literature review has identified 8 publications on SPG52, involving 18 patients from 12 pedigrees. Combined with our case, 14 had carried homozygous variants of the AP4S1 gene, 3 had compound heterozygous variants, and 2 had heterozygous variants, involving 12 distinct variant sites. The cohort included 7 males and 12 females. All patients exhibited progressive lower limb spasticity and weakness as the primary feature, with certain loss of independent ambulation. Most patients had intellectual disability, some had distinctive facial features, though febrile seizures or epilepsy were common. Electroencephalography often showed increased slow-wave activity. Brain MRI frequently demonstrated ventriculomegaly, a thin corpus callosum, and reduced white matter. Conclusion:The homozygous c. 289C>T (p.Arg97*) variant of the AP4S1 gene probably underlay the pathogenesis of SPG52 in this child. Above discovery has expanded the mutational spectrum of AP4S1 and provided valuable insights for the genetic diagnosis, counseling, and clinical management of SPG52.

AIM To explore the clinical effects of Jinfukang Oral Liquid combined with thymosin α1 on patients with non-small cell lung cancer due to Dual Deficiency of Qi and Yin.METHODS Seventy-five patients were randomly assigned into thymosin α1 group(15 cases)for 4-week administration,Jinfukang Oral Liquid group(30 cases)for 4-week administration,and combination group(30 cases)for 4-week administration.The changes in TCM clinical syndrome effects,immunity indices(CD3+T,Th,CTL,total NK,CD56dim CD16+NK,NKT,Treg,MDSC),lethality/inhibition ratios(CTL/Treg,total NK/Treg,NKT/Treg,CTL/MDSC,total NK/MDSC,NKT/MDSC)and FACT-L scores were detected.RESULTS The Jinfukang Oral Liquid group and combination group demonstrated higher total effective rates than the thymosin α1 group(P＜0.05).After the treatment,the Jinfukang Oral Liquid group and combination group displayed increased NKT(P＜0.05)and decreased MDSC(P＜0.05),which were more obvious than those in the thymosin α1 group(P＜0.05),and higher NKT was observable in the Jinfukang Oral Liquid group(P＜0.05);the Jinfukang Oral Liquid group and combination group displayed increased lethality/inhibition ratios(P＜0.05),among which NKT/Treg,CTL/MDSC,total NK/MDSC,NKT/MDSC were higher than those in the thymosin α1 group(P＜0.05),and higher CTL/MDSC,NKT/MDSC were observable in the Jinfukang Oral Liquid group(P＜0.05);the Jinfukang Oral Liquid group(except for physiological status,society and family status)and combination group(except for society and family status)displayed increased FACT-L scores(P＜0.05).CONCLUSION For the patients with non-small cell lung cancer due to Dual Deficiency of Qi and Yin,Jinfukang Oral Liquid single use or combined with thymosin α1 can enhance peripheral blood immune surveillance,inhibit immune escape,restore the balanced state of tumor immune responses,and improve TCM syndromes and life quality.

AIM To prepare fisetin-loaded nanostructured lipid carriers,and to evaluate their in vivo pharmacokinetics.METHODS Ethanol injection method was applied to preparing the nanostructured lipid carriers.With monostearin-phospholipid ratio,monostearin-triacetin ratio and D-α-tocopheryl polyethylene glycol 1000 succinate(TPGS)concentration as influencing factors,encapsulation efficiency as an evaluation index,the formulation was optimized by Box-Behnken response surface method.The crystal form was analyzed by X-ray powder diffraction,after which the morphology was observed by transmission electron microscopy,infrared spectroscopy analysis was performed,the drug relaese was investigated by dialysis bag method,and the stability was determined.Eighteen rats were randomly assigned into 3 groups and given intragastric administration of the 0.5％CMC-Na suspensions of fisetin and its phospholipid complex,nanostructured lipid carriers(150 mg/kg),respectively,after which blood collection was made at0.25,0.5,1,1.5,2,3,4,6,8,12 h,UPLC-MS/MS was adopted in the plasma concentration determination of fisetin,and main pharmacokinetic parameters were calculated.RESULTS The optimal formulation was determined to be 1.56∶1 for monostearin-phospholipid ratio,3.05∶1 for monostearin-triacetin ratio,and 0.2 mg/mL for TPGS concentration.The nearly round fisetin-loaded nanostructured lipid carriers demonstrated the average encapsulation efficiency,drug loading,particle size and Zeta potential of(86.14±1.28)％,(8.96±0.26)％,(212.35±9.04)nm and-(31.13±1.16)mV,respectively.Raw medicine existed in the nanostructured lipid carriers in an amorphous state,preparation process did not affect the hydrogen bonding between raw medicine and phospholipids.The nanostructured lipid carriers displayed the accumulative release rate of 46.12％within 3 h in simulated gastric juice,and that of about 50％within 18 h in simulated intestinal fluid,whose freeze-dried powder exhibited good stability after being placed for 6 months.Compared with raw medicine and phospholipids complex,the nanostructured lipid carriers displayed prolonged tmax,t1/2(P＜0.01)and increased Cmax,AUC0-t,AUC0-∞(P＜0.01),whose relative bioavailability was enhanced to 7.07 times.CONCLUSION Nanostructured lipid carriers can improve the oral bioavailability of fisetin.