Objective: This study employed next-generation sequencing (NGS) for large-scale genotyping of 42 blood group systems in blood donors to evaluate its applicability in multi-system blood group identification and rare blood type repository construction, while exploring the complementary value of third-generation sequencing (TGS) in haplotype phasing and variant capture in regions with insufficient sequencing depth for ABO ambiguous samples. Methods: A total of 562 regular blood donors (median donation frequency 11 times) from a blood center were enrolled. NGS was used for genotyping of 42 blood group systems, with a focused analysis on 12 systems essential for rare blood type repository construction: MNS, Lutheran, Kell, Duffy, Kidd, Diego, Yt, Colton, Gerbich, Ok, H, and I. TGS was employed to resolve haplotype phasing (i. e., determining whether different mutation sites are located on the same chromosome) in 12 ABO samples with discrepancies between forward and reverse typing. Serological and genotyping results were compared for MNS, Duffy, Kidd, Lewis, and ABO systems. Results: NGS-based genotyping revealed that Fy (a-b+) in the Duffy system accounted for 0.87% (4/458), and Di (a+b+) in the Diego system accounted for 10.62% (31/292). The Lutheran, Kell, Yt, Colton, H, Ok, I, and Gerbich systems exhibited near-uniform phenotype distributions. Among the 12 ABO ambiguous samples, NGS yielded multiple possible genotype combinations in 5 cases due to inability to phase alleles, whereas TGS uniquely resolved the genotypes through long-read sequencing. In one case, NGS detected only 2 mutation sites due to insufficient sequencing depth, while TGS identified all 11 sites and assigned the A1 phenotype. Concordance rates between serology and genotyping were: Duffy 96.55% (140/145), ABO 96.43% (513/532), Kidd 94.17% (97/103), MNS 85.07% (57/67), and Lewis 68.18% (75/110). In the MNS system, 7 of 8 samples serologically typed as M+N+ but genotyped as M-N+ carried GYPB variants. Lewis discrepancies predominantly featured genotype Le (a-b+) with serological phenotypes of Le (a+b-), Le (a+b+), or Le (a-b-). The NGS platform completed sequencing of 192 samples within 2 weeks. Conclusion: The tiered genotyping strategy combining NGS, TGS, and serology enables multi-system blood group identification in large-scale blood donor populations. TGS provides complementary value in phasing ambiguous ABO samples and detecting variants in regions with insufficient sequencing depth. This study provides a technical framework and baseline frequency data for the expansion of a local rare blood type repository. However, standardization and cost-effectiveness of this strategy require further validation with expanded sample sizes, and serological confirmation should be retained for secretion status-related systems such as Lewis.

Objective:To analyze differences in the expression of routine laboratory parameters and cerebrospinal fluid (CSF) examination indicators between patients with non-neurosyphilis (syphilis without nervous system involvement) and those with neurosyphilis, to screen for key predictive factors, and to construct a predictive model for neurosyphilis.Methods:A retrospective analysis was conducted on the clinical data from patients with syphilis at the Fifth People's Hospital of Suzhou from 2019 to 2024. Patients with neurosyphilis and non-neurosyphilis who were hospitalized from November 2019 to June 2022 were included in the model cohort, and those hospitalized from January 2024 to October 2024 were included in the validation cohort. The patients' basic information and laboratory test indicators (including routine blood tests, CSF biochemical analysis, and syphilitic antibody tests) were collected. Statistical analysis was performed using the GraphPad software. The receiver operating characteristic (ROC) curve and the binary logistic regression method were used to analyze the predictive performance of key indicators in patients from the model cohort with SPSS software, and a predictive model for neurosyphilis was constructed. The performance of the neurosyphilis predictive model for neurosyphilis was validated based on relevant indicators from the validation cohort.Results:The model cohort included 99 patients with non-neurosyphilis (including 49 males and 50 females), and they were aged between 19 and 85 years, with an average age of 47 years; 69 patients with neurosyphilis were also included in the model cohort, including 58 males and 11 females, and they were aged between 26 and 73 years, with an average age of 51 years. The neurosyphilis group showed a significant increase in the median levels of CSF adenosine deaminase (1 U/L) and microprotein (711 mg/L), white blood cell counts (0.009 × 10?/L), as well as in the proportion of positive Pandy tests (35/69, 50.7%) compared with the non-neurosyphilis group (0 U/L, 309 mg/L, 0.002 × 10?/L, 2 /99 [2.0%], respectively, all P < 0.001). Based on the ROC curve analysis, the CSF microprotein and white blood cell count had relatively high discriminative ability (area under the ROC curve [AUC] > 0.85), while adenosine deaminase and the Pandy test showed moderate discriminative ability (0.7 < AUC < 0.85). According to the above four indicators, the logistic regression analysis showed that CSF microprotein combined with CSF white blood cell counts could construct the best predictive model for neurosyphilis, with a prediction accuracy rate of 0.980, a sensitivity of 98.5%, and a specificity of 89.9%. The prediction formula was logit (p) = -9.926 + 0.015 × microprotein + 362.33 × CSF white blood cell count, with a cutoff value of ≥ -0.867. The validation cohort enrolled 72 patients with non-neurosyphilis and 51 with neurosyphilis, and there were significant differences in CSF microprotein levels and white blood cell counts between the two groups (both P < 0.001). In the validation cohort, the predictive model demonstrated an accuracy of 86.2%, with a sensitivity of 83.6% and a specificity of 91.1% for predicting neurosyphilis. Conclusion:The predictive model for neurosyphilis constructed by combining CSF microprotein and CSF white blood cell count may contribute to the early differential diagnosis of neurosyphilis.

Objective:The clinical symptoms of children with global developmental delay (GDD) were analyzed to provide the scientific basis for the intervention of children with GDD.Methods:The results of the neuro-psychobehavioral scale were collected from 32 511 children with GDD from June 2020 to November 2023. Inclusion criteria: Children diagnosed with GDD according to the Diagnostic and Statistical Manual of Mental Disorders-V, ages 0.0 to 4.9 years. Exclusion criteria: children with common hearing impairment and visual impairment. The Chi-square tests were used for statistical analysis.Results:There were more boys than girls with GDD in outpatient clinics (68.2% vs. 31.8%). Among the children, the proportion of developmental delay in 5, 4, 3, and 2 domains was 31.1%, 23.4%, 22.9% and 22.6% respectively. The rate of delay in 2-3 domains was lower in boys (41.9%) than in girls (53.1%). The rate of delay in 4-5 domains was higher in boys (58.1%) than in girls (46.9%) ( χ2=352.11, P<0.001). Overall, outpatient GDD decreased with age. From 1.0-1.9 to 4.0-4.9 years of age, the proportion of children with developmental delay in 5 domains increased with age (18.2%, 36.4%, 43.9%, 52.4%). Among children aged 0.0-0.9 years, the proportion of 2 domains of developmental delay was higher (33.4%).Among children aged 1.0-1.9 years, the proportion of 2-3 domains of developmental delay was higher (30.7%). Among children aged 2.0-, 3.0-, 4.0-4.9 years, the proportion of developmental delay in 5 domains was higher (36.4%, 43.9%, 52.4%). In children with GDD, the fine motor delay occurred most frequently (85.1%), followed by social self-care (83.9%), language (79.0%), adaptation (62.3%), and gross motor (52.8%). The frequency of developmental delays in fine motor, adaptability, language, and social self-care in boys was higher than that in girls ( χ2=161.37, χ2=41.10, χ2=320.90, χ2=238.54, all P<0.001). The age groups with the highest delay incidence of gross motor, fine motor, adaptability, language, and social self-care were: 4.0-4.9 years (70.6%), 3.0-3.9 years (97.4%), 4.0-4.9 years (81.2%), 2.0-2.9 years (90.9%),2.0-2.9 years (95.4%). The proportions of fine motor delay in GDD children aged 0.0-0.9, 3.0-3.9 and 4.0-4.9 years were (74.5%, 97.4%, 96.8%) and the proportions of social self-care delay in GDD children aged 1.0- and 2.0-2.9 years were (92.1%, 95.4%). Peripheral and mild developmental delays were predominant in children with GDD. The proportion of severe language delay (6.4%) was higher than that in other fields. Conclusions:The proportion of GDD children with developmental delay in 4-5 domains was 54.5%. The most frequent domain of delay was fine motor. The frequencies of developmental delays in fine motor skills, adaptability, language, and social self-care in boys were higher than in girls. Most of the developmental delays in GDD children were marginal and mild. The rate of severe developmental delay in language was higher than in other domains.

Objective:To analyze differences in the expression of routine laboratory parameters and cerebrospinal fluid (CSF) examination indicators between patients with non-neurosyphilis (syphilis without nervous system involvement) and those with neurosyphilis, to screen for key predictive factors, and to construct a predictive model for neurosyphilis.Methods:A retrospective analysis was conducted on the clinical data from patients with syphilis at the Fifth People's Hospital of Suzhou from 2019 to 2024. Patients with neurosyphilis and non-neurosyphilis who were hospitalized from November 2019 to June 2022 were included in the model cohort, and those hospitalized from January 2024 to October 2024 were included in the validation cohort. The patients' basic information and laboratory test indicators (including routine blood tests, CSF biochemical analysis, and syphilitic antibody tests) were collected. Statistical analysis was performed using the GraphPad software. The receiver operating characteristic (ROC) curve and the binary logistic regression method were used to analyze the predictive performance of key indicators in patients from the model cohort with SPSS software, and a predictive model for neurosyphilis was constructed. The performance of the neurosyphilis predictive model for neurosyphilis was validated based on relevant indicators from the validation cohort.Results:The model cohort included 99 patients with non-neurosyphilis (including 49 males and 50 females), and they were aged between 19 and 85 years, with an average age of 47 years; 69 patients with neurosyphilis were also included in the model cohort, including 58 males and 11 females, and they were aged between 26 and 73 years, with an average age of 51 years. The neurosyphilis group showed a significant increase in the median levels of CSF adenosine deaminase (1 U/L) and microprotein (711 mg/L), white blood cell counts (0.009 × 10?/L), as well as in the proportion of positive Pandy tests (35/69, 50.7%) compared with the non-neurosyphilis group (0 U/L, 309 mg/L, 0.002 × 10?/L, 2 /99 [2.0%], respectively, all P < 0.001). Based on the ROC curve analysis, the CSF microprotein and white blood cell count had relatively high discriminative ability (area under the ROC curve [AUC] > 0.85), while adenosine deaminase and the Pandy test showed moderate discriminative ability (0.7 < AUC < 0.85). According to the above four indicators, the logistic regression analysis showed that CSF microprotein combined with CSF white blood cell counts could construct the best predictive model for neurosyphilis, with a prediction accuracy rate of 0.980, a sensitivity of 98.5%, and a specificity of 89.9%. The prediction formula was logit (p) = -9.926 + 0.015 × microprotein + 362.33 × CSF white blood cell count, with a cutoff value of ≥ -0.867. The validation cohort enrolled 72 patients with non-neurosyphilis and 51 with neurosyphilis, and there were significant differences in CSF microprotein levels and white blood cell counts between the two groups (both P < 0.001). In the validation cohort, the predictive model demonstrated an accuracy of 86.2%, with a sensitivity of 83.6% and a specificity of 91.1% for predicting neurosyphilis. Conclusion:The predictive model for neurosyphilis constructed by combining CSF microprotein and CSF white blood cell count may contribute to the early differential diagnosis of neurosyphilis.

Objective To investigate the correlation between G protein-coupled receptor family C group 6 member A(GPRC6A)gene polymorphisms and their expression and pulmonary infections in elderly patients with chronic heart failure(CHF).Methods 138 elderly CHF patients admitted to the Xianyang First People's Hospital from January 2021 to January 2024 were selected as the research subjects,and were divided into an infected group(n=42)and an uninfected group(n=96)based on their lung infection status.Polymerase chain reaction(PCR)was used to detect polymorphisms at the rs6901250 and rs1606365 loci of the GPRC6A gene.The allele and genotype frequency distributions of the infected and uninfected groups were compared.Logistic regression modeling was used to analyze the s6901250 and rs1606365 loci under three genetic models(co-dominant,dominant and reces-sive)and lung infections in elderly patients with CHF.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression level of GPRC6A gene.The predictive value of the mRNA expression level of the GPRC6A gene for the development of pulmonary infections in elderly patients with CHF was analyzed by applying the receiver operator characteristic(ROC)curve.Results The distribution of genotypes at loci rs6901250 and rs1606365 of the GPRC6A gene in both the infected and uninfected groups of the lungs of elderly CHF patients conformed to the Hardy-Weinberg equilibrium law(χ2=0.199～0.376,all P>0.05),which was representative of the population.Compared with the uninfected group,the frequency of allele A at locus rs6901250(57.14%vs 41.67%)was significantly higher in the infected group,Allele G(54.76%vs.37.50%)and genotype GG(14.06%vs 29.99%)frequencies were significantly higher at locus rs1606365,and the differences were statistically significant(χ2=5.628,7.114,6.849,all P<0.05).At locus rs6901250,in the co-dominant model(GG vs AA)and the dominant model(GA+AA vs GG),the elderly CHF patients with AA genotype the risk of lung infection was higher than that of GG genotype(OR=1.753,1.546,all P<0.05);.rs1606365 locus showed that the risk of lung infection was higher than that of CC genotype in el-derly CHF patients with GG genotype under all three genetic models of co-dominant model(CC vs GG),dominant model(CG+GG vs CC)and recessive model(CG+CC vs GG)(OR=1.833,1.741,0.695,all P<0.05).The mRNA expression level of GPR-C6A gene in the lung-infected group of elderly CHF patients(1.43±0.35)was significantly higher than that in the uninfected group(1.02±0.21),and the difference was statistically significant(t=8.515,P<0.001).The results of the ROC curve analysis showed that the GPRC6A gene expression level predicted lung infection in elderly CHF patients with an AUC value of 0.895,a cut-offvalue of 1.37,and sensitivity and specificity of 85.7%and 66.7%,respectively.Conclusion The AA genotype at the rs6901250 locus and the GG genotype at the rs1606365 locus of the GPRC6A gene increased the risk of developing lung infec-tions in elderly patients with CHF.MRNA expression levels of the GPRC6A gene were elevated in the infected group,and its ex-pression level could be used as a predictive indicator for the development of lung infections in elderly patients with CHF.

AIM:To explore the therapeutic effects of Liuwei-Dihuang decoction(LD)on ovarian function in pre-mature ovarian failure(POF)mice,and to uncover the role of the Notch signaling pathway in its mechanism.METHODS:A mouse model of POF was constructed through intraperitoneal injection of cyclophosphamide/busulfan.Subsequently,the mice were randomly divided into six groups(n=12 per group):control,POF model,low-dose LD(LD-L),medium-dose LD(LD-M),high-dose LD(LD-H),and positive drug(0.7 mg/kg diethylstilbestrol)groups.The ovarian histopathologi-cal alterations were assessed by hematoxylin-eosin(HE)staining.Serum levels of follicle-stimulating hormone(FSH),anti-Müllerian hormone(AMH),and estradiol(E2)were quantified via ELISA assays.Immunohistochemistry(IHC)was conducted to compare the expression level of cell proliferation marker Ki-67 in mouse ovarian tissues among the groups.The expression of proteins involved in the Notch pathway components[Notch1,Notch2,Jagged1(JAG-1),and hairy and enhancer of split 1(Hes1)]was determined via IHC and Western blot,while the corresponding mRNA levels of Notch1,Notch2,JAG-1,JAG-2,Hes1,and Hes5 were quantified through the qPCR analysis.RESULTS:Ovarian weight was significantly higher,and the number of atretic follicles was significantly lower in the LD-H and positive drug groups com-pared to the model group(P<0.05).However,serum FSH levels were decreased(P<0.01),whereas those of E2 and AMH were elevated in both treatment groups compared with the model group(P<0.01).In the LD-H group,the expres-sion of key molecules of the Notch pathway components was increased at the protein and mRNA level compared with the model group(P<0.01).CONCLUSION:LD ameliorates ovarian dysfunction in POF mice via activating the Notch sig-naling pathway.

Keratoconus(KC)is a blinding eye disease caused by a variety of factors, with its pathogenesis still not well understood. In recent years, it has been discovered that sex hormones and prolactin-induced protein(PIP)have a profound impact on the cornea, with more noticeable changes when there are abnormalities in their body content. It has been found that various sex hormone receptors are distributed in the cornea. Based on this, this article reviews a multitude of studies on how sex hormones and PIP affect the cornea, along with relevant clinical research. It has been observed that sex hormones and PIP also play a role in KC patients and influence the occurrence and progression of KC. Additionally, it has been noted that pregnant and lactating women may be more susceptible to KC. Sex hormones and PIP have the potential to become new diagnostic and therapeutic targets. This article not only provides new insights but also offers important references for clinical practice.

Keratoconus(KC)is a blinding eye disease caused by a variety of factors, with its pathogenesis still not well understood. In recent years, it has been discovered that sex hormones and prolactin-induced protein(PIP)have a profound impact on the cornea, with more noticeable changes when there are abnormalities in their body content. It has been found that various sex hormone receptors are distributed in the cornea. Based on this, this article reviews a multitude of studies on how sex hormones and PIP affect the cornea, along with relevant clinical research. It has been observed that sex hormones and PIP also play a role in KC patients and influence the occurrence and progression of KC. Additionally, it has been noted that pregnant and lactating women may be more susceptible to KC. Sex hormones and PIP have the potential to become new diagnostic and therapeutic targets. This article not only provides new insights but also offers important references for clinical practice.

ChuanWu (CW), the dried mother root of Aconitum carmichaelii Debx., is a well-known traditional Chinese medicine (TCM) recognized for its potent efficacy but inherent toxicity, primarily due to its alkaloid content. Traditional and modern detoxification methods for CW include proper processing, rational compatibility, and specialized decoction techniques, among which honey-boiled CW is particularly distinctive. However, research on the detoxification mechanism of honey-boiled CW remains limited. This study investigated this mechanism by analyzing alkaloid transformation and supramolecular aggregation. Honey-boiled and water-boiled CW preparations were compared. Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to analyze CW alkaloids, specifically diester alkaloids (DDAs), monoester alkaloids (MDAs), and non-esterified diterpenoid alkaloids (NDAs). Transmission electron microscopy was employed to observe and identify supramolecular aggregates in the honey-boiled CW decoction. In vivo absorption of water-boiled, honey-boiled, and NADES-boiled CW was compared. Median lethal dose (LD₅₀) tests assessed toxicity, including hepatotoxicity and nephrotoxicity. In vitro experiments evaluated the safety, anti-inflammatory, and analgesic effects of CW-medicated serum on RAW264.7 cells, with in vivo validation in mice. Results showed that honey promoted the conversion of highly toxic DDAs to less toxic MDAs and prevented MDAs from hydrolyzing into NDAs. Honey-boiled CW formed approximately 250 nm supramolecular aggregates that encapsulated MDAs, inhibiting their conversion to NDAs. These encapsulated MDAs acted as a stable delivery system with higher bioavailability than free benzoylmesaconine. Subsequent mouse experiments confirmed that honey-boiled CW significantly increased the LD₅₀ of CW while reducing hepatotoxicity and nephrotoxicity. Additionally, honey-boiled CW significantly improved cell safety and enhanced anti-inflammatory and analgesic effects. Our findings reveal that honey-boiled CW exhibits a potent detoxification mechanism by influencing alkaloid transformation and facilitating the formation of supramolecular aggregates. This study lays the groundwork for developing detoxification or synergistic strategies within honey-boiled TCM.

Obstructive sleep apnea hypopnea syndrome (OSAHS) is a disease in which apnea and hypopnea occur during sleep, and the main symptoms are sleep snoring. OSAHS is not only closely related to the occurrence of various eye diseases, but also the severity of OSAHS can affect the progression of related eye diseases. At present, continuous positive airway pressure (CPAP) is an effective means to treat OSAHS, and the progression of related eye diseases has been improved correspondingly after CPAP treatment. In the future, it is necessary to further study the pathogenesis of OSAHS and enrich the research evidence of the association between OSAHS and eye diseases, so as to provide more comprehensive theoretical evidence for the prevention and treatment of OSAHS-related eye diseases.