To investigate the correlation between vitamin D nutritional status and insulin resistance in pubertal adolescents.

Liver transplantation (LT) has become a standard treatment for end-stage liver diseases, and graft injury is intricately associated with poor prognosis. Granzyme B (GZMB) plays a vital role in natural killer (NK) cell biology, but whether NK-derived GZMB affects graft injury remains elusive. Through the analysis of single-cell RNA-sequencing data obtained from human LT grafts and the isolation of lymphocytes from mouse livers following ischemia-reperfusion injury (IRI), we demonstrated that 2NK cells with high expression of GZMB are enriched in patients and mice. Both systemically and liver-targeted depletion of NK cells led to a notable reduction in GZMB⁺ cell infiltration, subsequently resulting in diminished graft injury. Notably, the reconstitution of Il2rg ^-/- Rag2 ^-/- mice with purified Gzmb-KO NK cells demonstrated superior outcomes compared to those with wild-type NK cells. Crucially, global knockout of GZMB and pharmacological inhibition exhibited remarkable improvements in liver function in both mouse IRI and rat LT models. Moreover, a phosphorylated derivative of FDA-approved vidarabine was identified as an effective inhibitor of mouse GZMB activity by molecular dynamics, which could provide a potential avenue for therapeutic intervention. Therefore, targeting NK cell-derived GZMB during the LT process suggests potential therapeutic strategies to improve post-transplant outcomes.

Bacillus velezensis DJ1 exhibits broad-spectrum antagonistic activity against diverse phytopathogenic fungi, while its biocontrol mechanisms against Fusarium graminearum, the causal agent of maize stalk rot, remain poorly characterized. In this study, we integrated genomics and transcriptomics to elucidate the antifungal mechanisms of strain DJ1. The results demonstrated that DJ1 inhibited F. graminearum with the efficacy of 64.4%, while its polyketide crude extract achieved the control efficacy of 55% in pot experiments against this disease. Whole-genome sequencing revealed a single circular chromosome (3 929 792 bp, GC content of 47%) harboring 12 biosynthetic gene clusters for secondary metabolites, six of which encoded known antimicrobial compounds (macrolactin H, bacillaene, difficidin, surfactin, fengycin, and bacilysin). Transcriptomic analysis identified 243 differentially expressed genes (152 upregulated and 91 downregulated, P < 0.05), which were potentially associated with the antagonistic activity against F. graminearum. KEGG enrichment analysis highlighted activation (P < 0.05) of cysteine/methionine metabolism, pentose phosphate pathway, and polyketide biosynthesis pathways, indicating that DJ1 employed synergistic strategies involving antimicrobial compound synthesis, energy metabolism enhancement, and nutrient competition to suppress pathogens. This study provides a theoretical foundation for developing novel microbial resources and application technologies to combat phytopathogenic fungi.
Fusarium/drug effects* ; Bacillus/metabolism* ; Plant Diseases/prevention & control* ; Antifungal Agents/pharmacology* ; Genomics ; Zea mays/microbiology* ; Transcriptome ; Gene Expression Profiling ; Antibiosis ; Multigene Family ; Multiomics

Objective:To discuss the effect of asiatic acid(AA)on the inflammation and oxidative stress damage induced by lipopolysaccharide(LPS)in the primary cultured hippocampus neurons,and to clarify its mechanism.Methods:The primary cultured rat hippocampus neurons(cell purity identified by immunofluorescence staining)were divided into control group,LPS(10 mg·L-1)group,and LPS+AA group(10 mg·L-1 LPS+10,20,and 40 μmol·L-1 AA),AA group(20 μmol·L-1 AA),ML385 group[10 μmol·L-1 nuclear factor erythroid 2-related factor(Nrf2)inhibitor],and LPS+ML385+AA group(10mg·L-1 LPS+10 μmol·L-1 ML385+20 μmol·L-1 AA).After drug treatment,methylthiazolyldiphenyl-tetrazolium bromide(MTT)method was used to detect the survival rates of the hippocampus neurons in various groups;lactate dehydrogenase(LDH)kit was used to detect the LDH leakage rates of the hippocampus neurons in various groups;enzyme linked immunosorbent assay(ELISA)kit was used to detect the expression levels of inflammatory factors[interleukin(IL)-1β and tumor necrosis factor(TNF)-α]and the activities of superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the hippocampus neurons in various groups;Griess method was used to detect the nitric oxide(NO)levels in supernatant of the hippocampus neurons in various groups;immunofluorescence staining was used to detect the expressions of Nrf2 and heme oxygenase-1(HO-1)proteins in the hippocampus neurons in various groups;Western blotting method was used to detect the expression levels of Nrf2,HO-1,nuclear factor-kappa B(NF-κB),and B-cell lymphoma 2(Bcl-2)proteins in the hippocampus neurons in various groups.Results:Compared with control group,the survival rate of the hippocampus neurons,SOD activity,and Bcl-2 expression level in the cells in LPS group were significantly decreased(P＜0.01),while the LDH leakage rate,expression levels of IL-1β and TNF-α,MDA level,and NO level,as well as the expression level of NF-κB protein,were significantly increased(P＜0.01);the fluorescence intensities and expression levels of Nrf2 and HO-1 proteins in hippocampus neurons were significantly decreased(P＜0.01).Compared with LPS group,the survival rates of hippocampus neurons,SOD activities,and expression levels of Bcl-2 in the cells in LPS+10 μmol·L-1 AA group and 20 μmol·L-1 AA group were significantly increased(P＜0.01),while the LDH leakage rates,expression levels of IL-1β and TNF-α,MDA levels,and NO levels,as well as expression levels of NF-κB protein,were significantly decreased(P＜0.05 or P＜0.01),and the fluorescence intensities and protein expression levels of Nrf2 and HO-1 in the cells were significantly increased(P＜0.01).Compared with LPS+20 μmol·L-1 AA group,the fluorescence intensities of Nrf2 and HO-1 in the cells in LPS+ML385+AA group were significantly decreased(P＜0.01),and the expression levels of Nrf2 protein in the nucleus and cytoplasm,the expression levels of HO-1 and Bcl-2 proteins in the cells were significantly decreased(P＜0.01),while the expression level of NF-κB protein was significantly increased(P＜0.01).Conclusion:AA can improve LPS-induced inflammation and oxidative stress damage in the primary cultured rat hippocampus neurons,and its mechanism may be related to the activation of the Nrf2/HO-1 signaling pathway.

Public health laboratory testing involves a wide range of sample types,complex matrices,diverse target analytes with varying concentrations,and multiple application contexts with different analytical requirements.As a critical step in public health laboratory analysis and testing,sample pretreatment plays a decisive role in ensuring the reproducibility and efficiency of the analytical methods.It directly affects the accuracy,sensitivity,and reliability of testing results,as well as the feasibility of downstream analyses.Traditional sample pretreatment techniques face persistent challenges,including low efficiency,limited throughput,restricted universal applicability,high organic solvent consumption,and poor compatibility with downstream analytical procedures.These limitations constrain their capacity to meet the evolving demands of research and practice in public health and preventive medicine.In recent years,technological advances have focused on improving efficiency and automation,enhancing selectivity and sensitivity,facilitating online testing capabilities,and promoting environmental sustainability.Sample pretreatment techniques in public health laboratory testing have been undergoing progressive upgrades,and numerous novel technologies have emerged.The paper provides a comprehensive review of new technologies and applications in the field.We focused on the development of new materials,the application of artificial intelligence,connections for online processing,and the approaches tailored to the demands of specific testing settings.We also discussed sample processing for omics analyses and mass spectrometry imaging methods relevant to public health laboratory testing.These advances are expected to support the development of greener and higher-throughput sample pretreatment and foster innovation in the public health laboratory testing system.

OBJECTIVE: To carry out genetic testing and clinical phenotypic characterization on a four-generation Chinese pedigree affected with Stickler syndrome type I and explore its genotype-phenotype correlation. METHODS: A child presented at the Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine in February 2023 for micrognathia, glossoptosis and cleft palate and his family members were selected as the study subjects. Clinical data were collected from the affected members, and peripheral blood samples were obtained from 17 participants (including 4 patients and 13 asymptomatic individuals). Whole exome sequencing (WES) was carried out. Candidate variant was verified by Sanger sequencing. Genotype-phenotype correlation was analyzed by integrating the sequencing data with evidence from existing literature. This study has bee granted by the Ethics Committee of Guangdong Provincial Hospital of Traditional Chinese Medicine and Guangzhou Women and Children's Medical Center (Ethics No.: 2022-406B00). RESULTS: The four-generation pedigree has comprised 19 members. In addition to the proband, 5 affected individuals had manifested with high myopia, congenital cataracts, and progressive vision loss. Two deceased members reportedly exhibited similar ocular manifestations. Among the four living patients, two had developed retinal detachment, while two others presented with chronic joint pain onset between 35 ~ 40 years of age. One patient required hip replacement surgery at age 42 secondary to femoral head necrosis. The proband, the youngest affected member, exhibited characteristic phenotypes including congenital micrognathia and cleft palate, consistent with Pierre-Robin syndrome. Genetic analysis revealed a heterozygous nonsense mutation in COL2A1 (NM_001844.5: c.2668C>T; p.Gln890Ter) segregating with the disease in all four symptomatic patients. This variant was absent in asymptomatic family members and unaffected controls. While the mutation is listed in ClinVar, no clinical case report has associated it with this phenotypic spectrum. It was not recorded in population databases (gnomAD v4.1.0, 1000 Genomes Project, or ExAC), supporting its potential pathogenicity. CONCLUSION This study has diagnosed a four-generation Chinese pedigree with Stickler syndrome type I attributed to the pathogenic COL2A1 variant c.2668C>T (p.Gln890Ter), which is a rare nonsense mutation associated with ocular predominance and variable skeletal involvement. Notably, this family exhibited marked clinical heterogeneity despite sharing the identical genotype, which highlighted the challenges in phenotype-genotype correlation. The autosomal dominant transmission pattern observed in this pedigree has provided critical insights into COL2A1-related collagenopathies and underscored the necessity of ultrasonographic monitoring for ocular anomalies during prenatal diagnosis. Above findings have advanced our understanding of the pleiotropic effects in type Ⅱ collagen disorders and laid the foundation for precision-based genetic counseling, enabling targeted cascade screening and implementation of tertiary prevention strategies against congenital disabilities for high-risk families.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Arthritis/genetics* ; Collagen Type II/genetics* ; Connective Tissue Diseases/genetics* ; Exome Sequencing ; Genetic Association Studies ; Genotype ; Hearing Loss, Sensorineural/genetics* ; Mutation ; Pedigree ; Phenotype ; Retinal Detachment/genetics* ; East Asian People/genetics*

In order to standardize the clinical diagnosis and treatment of upper airway cough syndrome (UACS) for children in China, Dongzhimen Hospital of Beijing University of Chinese Medicine and Affiliated Hospital of Liaoning University of Traditional Chinese Medicine initiated the development of this Traditional Chinese Medicine Diagnosis and Treatment Guidelines for Children with Upper Airway cough Syndrome based on evidence-based medical evidence. This guideline will process registration, write a plan, and develop relevant processes and writing norms, develop and publish official documents. This plan mainly introduces the scope of the guidelines, the purpose and significance, the composition of the guidelines working group, the management of conflicts of interest, the collection, selection and determination of clinical problems, the retrieval, screening and rating of evidence, and the consensus of recommendations. Registration information: This study has been registered in the international practice guidelines registry platform with the registration code of PREPARE-2023CN087.

Objective:To observe the effects of early energy intake and early enteral nutrition on prognosis of patients during extracorporeal membrane oxygenation (ECMO).Methods:Patients who received ECMO treatment in the emergency intensive care unit (EICU) of the Jiangsu Provincial Hospital (First Affiliated Hospital of Nanjing Medical University) from January 2021 to June 2022 were selected as subjects to summarize the early energy intake of ECMO patients. Logistic regression analysis and restricted cubic spline (RCS) analysis were used to determine the relationship between early energy intake and prognosis of ECMO patients. According to the results of RCS analysis, the patients were divided into energy-deficient group and energy-sufficient group. And according to whether early enteral nutrition (EEN) was initiated, the patients were divided into EEN group and non-EEN group. The differences of clinical outcomes between energy-deficient group and energy-sufficient group, EEN group and non-EEN group were compared.Results:There was no significant difference in age, sex, BMI, primary disease and ECMO pattern between energy-deficient group and energy-sufficient group, EEN group and non-EEN group. The ECMO conversion time (days) and hospitalization time (days) in the energy-deficient group were significantly lower than those in the energy-sufficient group, and the survival rate in the energy-deficient group was significantly lower than that in the energy-sufficient group [43.2% (19/44) vs. 66.0% (31/47), P=0.029]. Kaplan-Meier survival analysis showed that the 28-day survival rate in the energy-deficient group was significantly lower than that in the energy-sufficient group, and the risk of death was 2.595 times higher than that in the energy-sufficient group. The conversion time (days), hospital stay (days) and average daily energy intake [kcal/(kg·d)] in the EEN group were higher than those in the non-EEN group ( P<0.05), and the survival rate in the non-EEN group was significantly higher than that in the non-EEN group [66.1% (41/62) vs. 31.0% (9/29), P<0.002]. Kaplan-Meier survival analysis showed that the 28-day survival rate in the non-EEN group was significantly lower than that in the EEN group, and the risk of death was 2.981 times higher than that in the EEN group ( P<0.001). Conclusions:The energy intake of patients with ECMO above 16.94 kcal/ (kg·d) is a protective factor for prognosis. EEN helps to increase early energy intake and improve prognosis in patients during ECMO.

Objective:To analyze the relationship between systemic immune-inflammatory index (SII) and clinical characteristics of pancreatic cancer patients and the factors influencing SII.Methods:To retrospectively analyze the data of 98 pancreatic cancer patients with R 0 resection in the Department of Hepatobiliary Surgery, Affiliated Hospital of Guizhou Medical University, from January 2018 to January 2023, of whom 54 were male and 44 were female at the age of (59.2±10.7) years. All patients were divided into the high SII group ( n=49) and the low SII group ( n=49) based on the median SII of 668. Preoperative albumin, proportion of tumor located in the head of the pancreas, preoperative total bilirubin, and length of hospitalization were compared between the two groups. The correlation between SII and clinical features was analyzed. The SII influencing factors of pancreatic cancer patients were analyzed by linear regression with univariate and multivariate analysis. Results:The preoperative albumin of pancreatic cancer patients in the high SII group was lower than that of the low SII group, while the proportion of tumors located in the head of the pancreas, the preoperative total bilirubin, and the length of hospital stay were higher than that of the low SII group, and the differences were all statistically significant (all P<0.05). Correlation analysis showed a negative correlation between preoperative albumin and SII ( r=-0.28, P=0.050), while a positive correlation between preoperative total bilirubin and SII ( r=0.36, P<0.001), as well as between hospitalization time and SII ( r=0.28, P=0.050). The SII of patients with tumors located in the head of the pancreas ( n=81) and the tail of the pancreatic body ( n=17) were 734 (418, 1 241) and 431 (276, 613), respectively, and the SII of patients who survived ( n=92) and died ( n=4) at the time of discharge were 628 (383, 1 113) and 1 283 (1 176, 1 507), respectively, and the differences in SII of patients with different tumor locations and different discharge status were all significant. The SII of the patients were compared, and the differences were statistically significant ( Z=-2.51, 2.05, P=0.012, 0.038). Screening variables in the univariate linear regression model showed that admission total bilirubin, tumor volume, and preoperative albumin were associated with SII levels (all P<0.05). When the above variables were included in the multivariate linear regression, the higher the preoperative total bilirubin ( β=2.74, 95% CI: 1.48-4.00, P<0.001) as well as the larger the tumor volume ( β=2.34, 95% CI: 1.04-3.63, P<0.001) in pancreatic cancer patients, the higher the value of SII. Conclusions:SII in pancreatic cancer patients was associated with preoperative albumin, tumor location, preoperative total bilirubin, length of hospital stay, and survival or death at discharge. Preoperative total bilirubin and tumor volume were influential factors of SII in patients with pancreatic cancer.