ObjectiveThis study aims to explore the potential of different orders of magnitude single-nucleotide polymorphism (SNP) locus combinations for predicting distant kinship relationships. A high-density SNP locus set was constructed, and a comprehensive assessment of its inference capability was conducted. MethodsFirstly, we selected three commercial chip panels, CGA (Chinese genotyping array, Illumina), GSA (Global screening array, Illumina), Affy (23MF_V2 high-density SNP array, Affymetrix) and merged them after quality control, forming a high-density SNP locus panel(1 180 k). Secondly, we selected 161 samples and collected their peripheral blood samples by using whole-genome sequencing technology. Within this sample population, the levels of kinship relationships fully covered the range from level 1 to level 9, and the number of kinship pairs at each level was consistently maintained at over 50 pairs. From 161 samples data of whole-genome sequencing, the 1 180 k locus set was extracted, which is referred to as the high-density SNP locus set in the following text. The kinship inference was conducted using the identity-by-descent (IBD) algorithm with the selected optimal parameters. To comprehensively evaluate the performance of the high-density SNP locus set in kinship inference, we compared it with the three commercial chip panels, the intersection of these three chip loci, and the control sets constructed by randomly reducing the number of the high-density SNP locus set. Based on the changes in the IBD lengths, as well as the dynamic trends in prediction accuracy, we conducted a scientific assessment of the kinship inference capability of the high-density SNP locus set. ResultsAfter screening, a set of 1 184 334 autosomal SNPs was obtained. During the process of screening the optimal IBD length threshold, the result revealed that 0 cM, 1 cM, and 2 cM all demonstrated good applicability. However, to avoid the issue of a large amount of redundant information caused by setting a too low IBD length threshold, this study ultimately selected 2 cM as the optimal threshold. Compared with the average results of three chip panels, the high-density SNP locus set increased the total IBD length and the average IBD length across levels 1-9; the accuracy of the confidence interval for level 8 was 70.97%, which represented a 3.50% improvement; the average confidence interval accuracy for levels 1-8 was 91.39%, representing a 1.00% increase; and the false negative rates at levels 8 and 9 were reduced by 2.42% and 6.76%, respectively. The system efficacy of the high-density SNP locus set for kinship inference of first to eighth degree relationships reached 98.91%. Through random reduction of the high-density SNP locus set results, it is found that increasing the number of SNPs with the panel, the detection efficiency of IBD length showed a significant upward trend. At the same time, the overall trend in the accuracy of kinship relationship prediction as well as the confidence interval accuracy also indicated that both metrics steadily increased with the addition of more loci. ConclusionThe results show that the high-density SNPs panel significantly enhances the efficacy of distant kinship inference, accurately covering kinship degrees, with the average confidence interval accuracy for first to eighth degree relationships stably above 90%. The study finds that increasing the number of SNPs panel can improve the ability to predict distant kinship.

This study investigated the effect of Wuling San on transforming growth factor-β1(TGF-β1)-induced fibrosis, inflammation, and oxidative stress in human renal tubular epithelial cells(HK-2) and its mechanism of antioxidant stress injury. HK-2 cells were cultured in vitro and divided into a control group, a TGF-β1 model group, and three treatment groups receiving Wuling San-containing serum at low(2.5%), medium(5.0%), and high(10.0%) doses. TGF-β1 was used to establish the model in all groups except the control group. CCK-8 was used to analyze the effect of different concentrations of Wuling San on the activity of HK-2 cells with or without TGF-β1 stimulation. The expression of key fibrosis molecules, including actin alpha 2(Acta2), collagen type Ⅰ alpha 1 chain(Col1α1), collagen type Ⅲ alpha 1 chain(Col3α1), TIMP metallopeptidase inhibitor 1(Timp1), and fibronectin 1(Fn1), was detected using qPCR. The expression levels of inflammatory cytokines, including tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-8(IL-8), and interleukin-4(IL-4), were measured using ELISA kits. Glutathione peroxidase(GSH-Px), malondialdehyde(MDA), catalase(CAT), and superoxide dismutase(SOD) biochemical kits were used to analyze the effect of Wuling San on TGF-β1-induced oxidative stress injury in HK-2 cells, and the expression of nuclear factor E2-related factor 2(Nrf2), heme oxygenase 1(HO-1), and NAD(P)H quinone oxidoreductase 1(NQO1) was analyzed by qPCR and immunofluorescence. The CCK-8 results indicated that the optimal administration concentrations of Wuling San were 2.5%, 5.0%, and 10.0%. Compared with the control group, the TGF-β1 model group showed significantly increased levels of key fibrosis molecules(Acta2, Col1α1, Col3α1, Timp1, and Fn1) and inflammatory cytokines(TNF-α, IL-1β, IL-6, IL-8, and IL-4). In contrast, the Wuling San administration groups were able to dose-dependently inhibit the expression levels of key fibrosis molecules and inflammatory cytokines compared with the TGF-β1 model group. Wuling San significantly increased the activities of GSH-Px, CAT, and SOD enzymes in TGF-β1-stimulated HK-2 cells and significantly inhibited the level of MDA. Furthermore, compared with the control group, the TGF-β1 model group exhibited a significant reduction in the expression of Nrf2, HO-1, and NQO1 genes and proteins. After Wuling San intervention, the expression of Nrf2, HO-1, and NQO1 genes and proteins was significantly increased. Correlation analysis showed that antioxidant stress enzymes(GSH-Px, CAT, and SOD) and Nrf2 signaling were significantly negatively correlated with key fibrosis molecules and inflammatory cytokines in the TGF-β1-stimulated HK-2 cell model. In conclusion, Wuling San can inhibit TGF-β1-induced fibrosis in HK-2 cells by activating the Nrf2 signaling pathway, improving oxidative stress injury, and reducing inflammation.
Humans ; Oxidative Stress/drug effects* ; Transforming Growth Factor beta1/metabolism* ; Fibrosis/genetics* ; Cell Line ; Drugs, Chinese Herbal/pharmacology* ; Epithelial Cells/immunology* ; Inflammation/metabolism*

The differences in chemical quality characteristics between Xinjiang Rehmannia glutinosa and R. glutinosa were analyzed to provide a theoretical basis for the introduction and quality control of R. glutinosa. In this study, the high performance liquid chromatography(HPLC) fingerprints of 6 batches of Xinjiang R. glutinosa and 10 batches of R. glutinosa samples were established. The content of iridoid glycosides, phenylethanoid glycosides, monosaccharides, oligosaccharides, and polysaccharides in Xinjiang R. glutinosa and R. glutinosa was determined by high performance liquid chromatography-diode array detection(HPLC-DAD), high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD), and ultraviolet-visible spectroscopy(UV-Vis). The determination results were analyzed with by chemical pattern recognition and entropy weight TOPSIS method. The results showed that there were 19 common peaks in the HPLC fingerprints of the 16 batches of R. glutinosa, and catalpol, aucubin, rehmannioside D, rehmannioside A, hydroxytyrosol, leonuride, salidroside, cistanoside A, and verbascoside were identified. Hierarchical cluster analysis(HCA) and principal component analysis(PCA) showed that Qinyang R. glutinosa, Mengzhou R. glutinosa, and Xinjiang R. glutinosa were grouped into three different categories, and eight common components causing the chemical quality difference between Xinjiang R. glutinosa and R. glutinosa in Mengzhou and Qinyang of Henan province were screened out by orthogonal partial least squares discriminant analysis(OPLS-DA). The results of content determination showed that there were glucose, sucrose, raffinose, stachyose, polysaccharides, and nine glycosides in Xinjiang R. glutinosa and R. glutinosa samples, and the content of catalpol, rehmannioside A, leonuride, cistanoside A, verbascoside, sucrose, and glucose was significantly different between Xinjiang R. glutinosa and R. glutinosa. The analysis with entropy weight TOPSIS method showed that the comprehensive quality of R. glutinosa in Mengzhou and Qinyang of Henan province was better than that of Xinjiang R. glutinosa. In conclusion, the types of main chemical components of R. glutinosa and Xinjiang R. glutinosa were the same, but their content was different. The chemical quality of R. glutinosa was better than Xinjiang R. glutinosa, and other components in R. glutinosa from two producing areas and their effects need further study.
Rehmannia/classification* ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Quality Control

OBJECTIVES: To study the impact of family socioeconomic status on children's reading fluency and the chain mediation effect of family reading environment and children's living and learning styles in this relationship. METHODS: A total of 473 children from grades 2 to 6 in two primary schools in Nanjing were selected through stratified random sampling. The children's reading fluency was assessed, and a questionnaire was used to collect information on family socioeconomic status, family reading environment, and children's living and learning styles. The mediation model was established using the Process macro in SPSS, and the Bootstrap method was employed to test the significance of the mediation effects. RESULTS: Family socioeconomic status, family reading environment, and children's living and learning styles were significantly positively correlated with reading fluency (P<0.001). The family reading environment and children's living and learning styles mediated the relationship between family socioeconomic status and children's reading fluency. Specifically, the independent mediation effect of family reading environment accounted for 11.02% of the total effect, while the independent mediation effect of children's living and learning styles accounted for 10.79%. The chain mediation effect of family reading environment and children's living and learning styles accounted for 7.41% of the total effect. CONCLUSIONS Family socioeconomic status can affect children's reading fluency through three pathways: family reading environment, children's living and learning styles, and the chain mediation effect of family reading environment and children's living and learning styles.
Humans ; Child ; Male ; Female ; Reading ; Learning ; Social Class ; Family

Objective To analyze the results of ultrasonic examinations of hip joints in preterm infants,clarify the developmental characteristics of hip joints across different gestational ages.Meth-ods A total of 881 preterm infants who attended the Child Healthcare Outpatient Clinic of Nanjing Maternity and Child Health Care Hospital between January 2023 and April 2024 were enrolled.The first ultrasonic examination of the hip joints was performed at a corrected gestational age of 4 to 6 weeks.The Graf classification method was employed to categorize the ultrasonic findings.Differences in hip joint development among preterm infants of varying gestational ages were compared,and rele-vant influencing factors were statistically analyzed.Results A total of 1,762 hip joints were exam-ined,with 30 cases of abnormal hip joints detected,yielding an abnormality detection rate of 3.4％(30/881).Among these,26 hips were classified as type Ⅱa and 8 as type Ⅱb.Statistically signifi-cant differences were observed in the detection of left-sided abnormal hip joints between preterm in-fants of other gestational age groups and late preterm infants(P＜0.05).A statistically significant difference was found in the classification of right-sided hip joints between preterm infants of different genders(P＜0.05),while no significant difference was observed for left-sided hip joints(P＞0.05).Statistically significant differences were noted in the detection rates of bilateral abnormal hip joints among preterm infants with varying birth weights(P＜0.05).Advanced maternal age was found to influence the development of left-sided hip joints inpreterm infants,whereas factors such as assisted reproductive technology,breech presentation,and nulliparity had no significant impacts on abnormal hip joint development in preterm infants.Conclusion Ultrasonic technology demonstrates signifi-cant advantages in screening for developmental dysplasia of the hip(DDH)in preterm infants.Dif-ferent gestational ages exert a notable influence on the development of left-sided hip joints in preterm infants.Female preterm infants exhibit a higher risk of right-sided hip joint abnormalities compared to the left side.Breech presentation,assisted reproductive technology,and nulliparity are not risk factors for DDH in preterm infants.However,higher birth weight in preterm infants and advanced maternal age are associated with abnormal hip joint development.Clinical attention should focus on high-risk factors,and enhanced dynamic ultrasonic monitoring of hip joints in preterm infants is war-ranted to facilitate early intervention and treatment.

Objective:To investigate whether sivelestat sodium (SV) mitigates paraquat (PQ)-induced acute lung injury (ALI)/acute respiratory distress syndrome (ARDS)-associated pulmonary fibrosis in mice by suppressing the NLRP3 inflammasome signaling pathway.Methods:Male C57BL/6J mice were randomly divided into four groups ( n=8 per group): Control group, PQ group, PQ+SV group, and SV group. The PQ and PQ+SV groups received an intraperitoneal injection of PQ solution (20 mg/kg) to establish a PQ poisoning model, while the Control and SV groups received an equivalent volume of saline. One hour later, the PQ+SV and SV groups were administered SV solution (100 mg/kg) intraperitoneally, whereas the Control and PQ groups received saline. After 48 hours, the mice were euthanized, and lung tissues were collected. Pathological changes were assessed via hematoxylin-eosin (HE) and Masson staining, followed by Smith and Ashcroft scoring. Immunohistochemistry was performed to evaluate the expression of NLRP3 inflammasome, caspase-1, α-smooth muscle actin (α-SMA), and collagen I. Western blotting was used to measure NLRP3 protein levels. Intergroup comparisons were conducted using one-way ANOVA with LSD post-hoc correction. Results:The Control and SV groups exhibited normal lung morphology, whereas the PQ+SV group showed reduced hemorrhage, congestion, and edema compared to the PQ group. Both PQ and PQ+SV groups exhibited significant weight loss post-intervention compared to the Control group (both P<0.001). HE and Masson staining revealed thickened alveolar septa, congestive and edematous alveolar walls, extensive inflammatory cell infiltration, and collagen deposition in the PQ group. In contrast, the PQ+SV group demonstrated alleviated alveolar wall congestion, reduced inflammatory infiltration, and decreased collagen deposition, with significantly lower Smith and Ashcroft scores [(5.92±1.34) vs. (10.88±1.88), P<0.001; (3.42±1.35) vs. (5.75±0.79), P<0.001]. Immunohistochemistry indicated reduced expression percentages of NLRP3 and caspase-1 in the PQ+SV group compared to the PQ group [(12.79%±0.43%) vs. (16.59%±0.40%), P<0.001; (17.71%±0.92%) vs. (19.84%±0.71%), P<0.001]. Similarly, α-SMA and collagen I expression in lung interstitium was significantly lower in the PQ+SV group [(11.79%±0.58%) vs. (16.14%±0.74%), P<0.001; (16.43%±0.56%) vs. (18.86%±0.60%), P<0.001]. Western blotting confirmed decreased NLRP3 protein expression in the PQ+SV group [(0.54±0.12) vs. (0.81±0.24), P<0.05]. Conclusions:SV attenuates PQ-induced ALI/ARDS-associated pulmonary fibrosis progression by inhibiting the NLRP3 inflammasome, suggesting that NLRP3 may be a key therapeutic target for early intervention in PQ-induced pulmonary fibrosis.

Intricate and complex interactions between tumor cells and tumor microenvironment(TME)are essential driving factors in tumor development and metastasis.In recent years,modulation of TME to reverse tumor progression has garnered significant attention as a therapeutic strategy.For precise and targeted tumor treatment,there is an urgent need for accurate and effective TME tar-get molecules and relevant prediction model.Fibronectin,a protein with multiple domains in extracellular matrix(ECM),plays a cru-cial role in physiological and pathological processes.Particularly,Fibronectin extra domain B(FN-EDB)in fibronectin is pivotal in regulating interaction between ECM and cells.During active tissue remodeling processes,especially in angiogenesis,FN-EDB expres-sion tends to increase.Studies have indicated that FN-EDB is involved in regulating various key aspects of tumorigenesis,including promoting tumor cell invasion and metastasis,influencing development of immune evasion mechanisms,enhancing tumor angiogene-sis,and contributing to development of chemoresistance.Therefore,targeting FN-EDB as a potential therapeutic target for cancer treat-ment and developing relevant therapeutic drugs hold significant importance and promising clinical prospects.This article reviews roles of FN-EDB in TME in recent years and research progress on tumor immunotherapy targeting this molecule,with aim of providing new insights and strategies for cancer treatment.

Intricate and complex interactions between tumor cells and tumor microenvironment(TME)are essential driving factors in tumor development and metastasis.In recent years,modulation of TME to reverse tumor progression has garnered significant attention as a therapeutic strategy.For precise and targeted tumor treatment,there is an urgent need for accurate and effective TME tar-get molecules and relevant prediction model.Fibronectin,a protein with multiple domains in extracellular matrix(ECM),plays a cru-cial role in physiological and pathological processes.Particularly,Fibronectin extra domain B(FN-EDB)in fibronectin is pivotal in regulating interaction between ECM and cells.During active tissue remodeling processes,especially in angiogenesis,FN-EDB expres-sion tends to increase.Studies have indicated that FN-EDB is involved in regulating various key aspects of tumorigenesis,including promoting tumor cell invasion and metastasis,influencing development of immune evasion mechanisms,enhancing tumor angiogene-sis,and contributing to development of chemoresistance.Therefore,targeting FN-EDB as a potential therapeutic target for cancer treat-ment and developing relevant therapeutic drugs hold significant importance and promising clinical prospects.This article reviews roles of FN-EDB in TME in recent years and research progress on tumor immunotherapy targeting this molecule,with aim of providing new insights and strategies for cancer treatment.

Objective This study aimed to investigate the potential relationship between urinary metals copper (Cu), arsenic (As), strontium (Sr), barium (Ba), iron (Fe), lead (Pb) and manganese (Mn) and grip strength. Methods We used linear regression models, quantile g-computation and Bayesian kernel machine regression (BKMR) to assess the relationship between metals and grip strength.Results In the multimetal linear regression, Cu (β=-2.119), As (β=-1.318), Sr (β=-2.480), Ba (β=0.781), Fe (β= 1.130) and Mn (β=-0.404) were significantly correlated with grip strength (P < 0.05). The results of the quantile g-computation showed that the risk of occurrence of grip strength reduction was -1.007 (95％ confidence interval:-1.362, -0.652; P < 0.001) when each quartile of the mixture of the seven metals was increased. Bayesian kernel function regression model analysis showed that mixtures of the seven metals had a negative overall effect on grip strength, with Cu, As and Sr being negatively associated with grip strength levels. In the total population, potential interactions were observed between As and Mn and between Cu and Mn (Pinteractions of 0.003 and 0.018, respectively).Conclusion In summary, this study suggests that combined exposure to metal mixtures is negatively associated with grip strength. Cu, Sr and As were negatively correlated with grip strength levels, and there were potential interactions between As and Mn and between Cu and Mn.

Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.