Huijuan Wang

Background: Hyperphenylalaninemia (HPA) exhibits distinct regional and ethnic heterogeneity in both genotypic profiles and phenotypic manifestations. However, as a region characterized by multi-ethnic habitation and profound cultural exchange, there remains a paucity of data regarding the genotype-phenotype correlation in patients from Xinjiang. This study aims to characterize the HPA-related gene mutation spectrum and phenotypic landscape in neonates with HPA from Xinjiang, evaluate genotype-phenotype correlation, and establish an evidence base for precise diagnosis and individualized clinical intervention. Results: A total of 273 patients with HPA were enrolled in this study, comprising 150 Han, 42 Hui, 36 Uyghur, 15 Kazakh, 4 Mongolian, 3 Dongxiang, and 23 individuals of unavailable ethnic origin. Next-generation sequencing was employed to identify variants in the phenylalanine hydroxylase ( PAH ) gene and Tetrahydrobiopterin (BH ₄ ) metabolic pathway genes [6-pyruvoyltetrahydropterin synthase ( PTS ), GTP cyclohydrolase 1 ( GCH1 ), quinoid dihydropteridine reductase ( QDPR )]. Putative pathogenic variants were validated via Sanger sequencing in trios. These genetic data were integrated with clinical phenotypes to perform association analysis using Genotype-Phenotype Value ( GPV ). A total of 101 distinct variants were identified in 273 patients, with missense mutations being predominant (71.29%, 72/101). The most frequent PAH mutations were c.158G>A (p.Arg53His) (15.38%, 84/546) and c.728G>A (p.Arg243Gln) (10.07%, 55/546). Notably, c.158G>A was highly enriched in Hui, Uyghur, and Kazakh populations, while ethnic-specific variants such as c.688G>A in the Uyghur population were also observed. Compound heterozygosity was the primary genotypic pattern (63.98%), and mild HPA was the most common phenotype (64.10%). Genotype-based phenotype prediction achieved accuracies of 98.78% for mild hyperphenylalaninemia, 46.15% for mild PKU and 81.48% for classical PKU. Prediction accuracy reached 100% for homozygous mutations and 88.29% for heterozygous mutations. Furthermore, GPV was significantly negatively correlated with blood phenylalanine levels ( r =-0.905, R ² =0.819, p =0.001). Conclusions: The genetic landscape of HPA in Xinjiang exhibits distinct regional features driven by unique ethnic genetic backgrounds. The high-frequency mild mutation c.158G>A in the PAH gene, distributed across multiple ethnicities, along with other ethnic-specific variants constitutes a region-specific mutation spectrum. This profile dictates clinical severity through the interplay of variant type and compound heterozygosity. Our findings confirm the clinical utility of the GPV in these multi-ethnic cohorts, and expand the global PAH mutation database.

Background: Long-term pharmacotherapy is the primary approach for alleviating the symptoms of mental disorders. Numerous pharmacogenomic studies have confirmed that genetic polymorphisms are key factors underlying inter-ethnic and inter-individual differences in drug response. Pharmacogenomic testing is an essential pathway to achieve precision medicine. Methods: In this study, seven common single nucleotide polymorphisms (SNPs) closely associated with the efficacy of antidepressants, and antipsychotics were screened and identified through comprehensive literature retrieval and mining of authoritative databases. These SNPs are located in the ANKK1, DRD2, FKBP5, GRIK4, HTR1A, HTR2A, and MC4R genes. Utilizing multiplex fluorescence melting curve analysis (mFMCA), a four-channel multiplex fluorescent PCR system was successfully established, enabling the simultaneous genotyping of 14 distinct alleles across 7 SNP loci in a single tube. Analytical performance including sensitivity and specificity as well as reproducibility was rigorously evaluated. Specifically, a two phase validation was conducted. An initial calibration phase using 98 clinical samples within a training cohort was used to establish integrative genotyping criteria. These criteria combined statistical melting temperature (T_m) intervals, defined as the Mean T_m± 3 standard deviations (SDs), with peak morphology. The established criteria were then evaluated in a blinded validation phase using an independent cohort of 124 clinical samples. Results: The mFMCA system can accurately detect genomic DNA at concentrations as low as 100 copies/μL, and exhibits high specificity and no cross-reactivity. In the validation cohort (n=124), the mFMCA-derived genotypes achieved 100% concordance with Sanger sequencing. Furthermore, the high stability of the system was further evidenced by its consistent performance across varying DNA concentrations, different clinical samples, and multiple instruments. Conclusion: This study successfully established a dedicated and cost-effective mFMCA assay for psychotropic pharmacogenomic testing. The method provides an efficient, accurate, rapid, economical tool for personalized medicine in psychiatric care.

The rising resistance of pathogens has increasingly complicated antibiotic treatments, leading to prolonged therapy durations and the potential for inadequate disease management. Current clinical methods (e.g., culture-based AST, conventional PCR, Sanger sequencing) for assessing microbial resistance are hampered by lengthy processing times, limited sensitivity, or high costs. In this study, we developed a multiplex quantitative polymerase chain reaction (qPCR) method using custom-designed universal locked nucleic acid (LNA) probes. This approach eliminates the need for target-specific fluorescent probes required in conventional qPCR assays, thereby offering broad applicability and reduced development costs. Using this method, a multiplex qPCR assay consisting of four tubes of four-plex reaction for the detection of 16 common antibiotic resistance genes (ARGs) was developed. These ARGs correspond to seven types of commonly used antibiotics and show high concordance with phenotypic resistance profiles. The established multiplex system demonstrated robust amplification performance, with a detection limit of 25 copies/μL for all targets. The analytical performance of this assay was validated in 172 samples from patients with severe pneumonia, achieving complete concordance with Sanger sequencing. Overall, this study provides an accurate, cost-effective, and rapid solution for detecting microbial resistance, enabling timely informed selection of antibiotics in clinical practice.

The main treatment strategy for bacterial vaginosis (BV) is antibiotic therapy, but they are not effective in restoring the vaginal microecology, which can lead to recurrence. This study aimed to isolate the dominant lactobacilli strains from the vagina of healthy Chinese women, evaluate their potential probiotic properties and explore their alleviative effect on BV infected mice. Our study isolated 43 strains of vaginal-dominant Lactobacillus. Four strains of Lactobacillus. crispatus, four strains of Lactobacillus. gasseri, and two strains of Lactobacillus. jensenii showed strong inhibitory effects on Gardnerella vaginalis and were safe in vitro. Subsequently, through principal component analysis on seven functional indicators of these 10 strains, Lact. crispatus G-7, Lact. gasseri B-5, and Lact. jensenii M-1 ranked the top beneficial strains and were finally selected as the candidate therapeutic strain for in vivo experiments. Animal experiments showed that Lact. crispatus G-7 was superior to other Lactobacillus species and the positive control metronidazole in restoring vaginal health (pH and H₂O₂). Additionally, it also significantly reduced the expression of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α while increasing the expression of anti-inflammatory cytokine IL-10. H&E staining and 16S rRNA gene sequencing showed that Lact. crispatus G-7 can alleviate the inflammatory damage caused by G. vaginalis and restored vaginal microecology. Among the isolated dominant Lactobacillus strains from the vagina, Lact. crispatus G-7 has been identified as the most effective in the treatment of G. vaginalis imparted BV. This strain is therefore a candidate for developing promising treatments for BV.

Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor. Notwithstanding tremendous efforts having been put in multi-omics research to profile the dysregulated molecular mechanisms and cellular pathways, there is still a lack of understanding about the glycoproteomic of GBM. Glycosylation as one of the most important post-translational modifications is crucial in regulating cell proliferation and relevant oncogenic pathways. In the study, we systematically profiled N-glycoproteomics of para-cancerous and cancerous tissues from GBM patients to reveal the site-specific N-glycosylation pattern defined by intact glycopeptides. We identified and quantified 1863 distinct intact glycopeptides (IGPs) with 161 N-linked glycan compositions and 326 glycosites. There were 396 IGPs from 43 glycoproteins differed between adjacent tissues and GBM. Then, proteomic and glycoproteomic data were combined, and the normalized glycosylation alteration was calculated to determine whether the difference was attributed to the global protein levels or glycosylation. The altered glycosylation triggered by site-specific N-glycans and glycoprotein abundance, as well as glycosite heterogeneity, were demonstrated. Ultimately, an examination of the overall glycosylation levels revealed a positive contribution of sialylated or/and fucosylated glycans. Overall, the dataset highlighted molecular complexity and distinct profiling at translational and post-translational levels, providing valuable information for novel therapeutic approaches and specific detection strategies.

Glioma is the most common malignant brain tumor. Previous studies have reported that calnexin (CANX) is significantly up-regulated in a variety of malignant tumors, including glioma, but its biological function and mechanism in glioma is still unclear. In this study, differentially expressed proteins in 3 primary glioblastoma multiforme (GBM) tissues and 3 paracancer tissues were identified by liquid chromatography-tandem mass spectrometry-based proteomic and bioinformatic analysis. The biological function and molecular mechanism of CANX were studied in glioma cell lines (T98G and A172) by CCK-8 assay, matrigel invasion assay, wound healing assay, flow cytometry and so on. Bioinformatics methods were used to analyze the immune microenvironment of glioma patients in TCGA database, and single cell sequencing data of 8 cases of untreated primary GBM in GEO database were analyzed. Proteomic analysis found that CANX was significantly overexpressed in glioma tissues comparing with paracancer tissues. The data from TCGA validated this result and showed that CANX was associated with poor prognosis of patients. A series of experiments at the cellular level found that CANX overexpression significantly enhanced the proliferation, migration and invasion ability of GBM cells, whereas CANX silencing had opposite effects. Further research found this effect may be mediated through the activation of the PI3K/AKT/mTOR signaling. In addition, immune infiltration analysis found that CANX high-expression glioma tissues exhibited fewer CD8⁺ T cells, natural killer cells and mast cells, along with significantly decreased tumor purity and significantly increased immune checkpoints expression. Single cell sequencing data analysis indicated that CANX was primarily expressed in astrocytes and dendritic cells. In conclusion, this study suggested that CANX may promote the malignant progression of glioma through PI3K/AKT/mTOR signaling pathway and play an important role in glioma immune escape. Therefore, CANX may be a valuable therapeutic target for glioma.

Background: This study aims to determine the efficacy and safety profile of aumolertinib in the real-word treatment setting for advanced non-small-cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) mutations.Methods: We retrospectively analyzed the clinical data of 173 EGFR-mutated advanced NSCLC patients who received aumolertinib treatment at Henan Cancer Hospital from April 2020 to December 2022. Progression-free survival (PFS) and overall survival (OS) were evaluated using Kaplan–Meier survival curves, while a Cox regression model was used for multifactorial analysis and prognostic factor assessment.Results: Among patients administered first-line aumolertinib (n = 77), the objective remission rate (ORR) of 77.92% was observed, along with a disease control rate (DCR) of 100%. The median progression-free survival (mPFS) was 24.97 months, which did not reach the median overall survival (mOS). The patients treated with aumolertinib after progression on prior EGFR-tyrosine kinase inhibitor (TKI) therapy (n = 96) exhibited an ORR of 46.88%, a DCR of 89.58%, an mPFS of 15.17 months, and an mOS of 21.27 months. First-line treatment multivariate Cox regression analysis demonstrated a statistically significant impact of elevated creatine kinase on PFS (p = 0.016) and a similar significant influence of co-mutation on OS (p = 0.034). Furthermore, subsequent-line treatment multivariate Cox regression analysis showed a statistically significant impact of elevated creatine kinase on median PFS (p = 0.026) and a significant effect on the number of metastatic organs (p = 0.017), co-mutation (p = 0.035), and elevated creatine kinase (p = 0.014) on median OS.Conclusion: Aumolertinib has shown clinical significance and can safely be used in the real-world setting for patients with EGFR mutation-positive NSCLC.

Purpose: Glioblastoma (GBM) is the most common and aggressive primary brain tumor characterized by poor prognosis and high recurrence. The underlying molecular mechanism that drives tumor progression and recurrence is unclear. This study is intended to look for molecular and biological changes that play a key role in GBM recurrence.Experimental design: An integrative transcriptomic and proteomic analysis was performed on three primary GBM and three recurrent GBM tissues. Omics analyses were conducted using label‐free quantitative proteomics and whole transcriptome sequencing.Results: A significant difference was found between primary GBM and recurrent GBM at the transcriptional level. Similar to other omics studies of cancer, a weak overlap was observed between transcriptome and proteome, and Procollagen C‐Endopeptidase Enhancer 2 (PCOLCE2) was observed to be upregulated at mRNA and protein levels. Analysis of public cancer database revealed that high expression of PCOLCE2 is associated with poor prognosis of patients with GBM and that it may be a potential prognostic indicator. Functional and environmental enrichment analyses revealed significantly altered signaling pathways related to energy metabolism, including mitochondrial ATP synthesis‐coupled electron transport and oxidative phosphorylation.Conclusions and clinical relevance: This study provides new insights into the recurrence process of GBM through combined transcriptomic and proteomic analyses, complementing the existing GBM transcriptomic and proteomic data and suggesting that integrated multi‐omics analyses may reveal new disease features of GBM.