Objective To verify the double antibody sandwich ELISA method for the detection of residual recombinant trypsin(RT) in monovalent virus harvest fluid and monovalent bulk solution of influenza split virion vaccine(MDCK cells).Methods Double antibody sandwich ELISA was used to detect residual levels of RT in monovalent virus harvest fluid and monovalent bulk solution, and was verified for the limit of quantitation(LOQ), specificity, repeatability, intermediate precision, accuracy and robustness.Results The method exhibited good linearity within the range of 10-0. 156 ng/mL, with a correlation coefficient R~2 of 1. 000. The LOQ was 0. 156 ng/mL. After diluting the monovalent harvest by a factor of two using sample diluent and phosphate buffer, the coefficient of variation(CV) for RT content was 1%, and the RT content measured after diluting monovalent bulk solution two-fold with sample diluent and phosphate buffer was less than 0. 156 ng/mL.The spiked recovery rates of RT solutions at different concentrations ranged from 81% to 105%, with a CV ranging from 1%to 6%. The CV of RT content measured by different test personnel in monovalent harvest was 3%, and the RT content in the monovalent bulk solution measured by different operators was also less than 0. 156 ng/mL. The CV of RT residue content in monovalent harvest at different chromogenic times was 5%, and the residual RT content in the monovalent bulk solution was less than 0. 156 ng/mL, indicating this detection method had good tolerance to different chromogenic times.Conclusion The double antibody sandwich ELISA method for detecting residual RT in monovalent virus harvest solution and monovalent bulk solution of influenza split virion vaccine(MDCK cells) has good linearity and range, LOQ, specificity, repeatability, intermediate precision, accuracy and robustness, and is suitable for the detection of residual RT in influenza split virion vaccine(MDCK cells).

ObjectiveTo investigate the impact of melanoma-associated antigen D4 （MAGED4） on the malignant biological behavior of hepatocellular carcinoma （HCC）， providing experimental evidence for its evaluation as a potential therapeutic target in HCC. MethodsGenes positively correlated with MAGED4 expression in HCC were screened using the LinkedOmics database， followed by GO functional and KEGG pathway enrichment analyses. Endogenously low-expressing SMMC-7721 and Huh7 cells were selected， with normal hepatic LO2 cells serving as controls. Stable overexpression cell lines were established via lentiviral infection （MAGED4 overexpression group）， with RT-PCR and Western blot validating overexpression efficiency. The effects of MAGED4 overexpression on cell proliferation， cell cycle distribution， migration， and adhesion were systematically analyzed using CCK-8 assays， flow cytometry， scratch healing assays， Transwell migration assays， and cell adhesion assays. ResultsBioinformatics analysis indicated that MAGED4 co-expressed genes were significantly enriched in biological processes closely associated with tumor proliferation and metastasis， including cell-cell adhesion， extracellular matrix organization， and the PI3K-Akt signaling pathway. A stable MAGED4 overexpression cell model was successfully established. In vitro functional assays demonstrated that MAGED4-overexpressing SMMC-7721 and Huh7 cells exhibited a mesenchymal-like morphology， whereas LO2 cells maintained an epithelial-like morphology. Compared with the empty-virus control groups， the MAGED4-overexpressing groups showed significantly increased proliferative activity （48 h： t=3.887， 22.070， 8.866， all P<0.05； 72 h： t=3.231， 23.840， 9.848， all P<0.05； 96 h： t=5.123， 49.800， 5.909， P<0.05， P<0.01， P<0.05）， wound healing rates （12 h： F=40.90， 44.00， 200.50， all P<0.001； 24 h： F=112.00， 34.89， 249.20， all P<0.001）， and numbers of migrated cells （F=239.20， 59.79， 132.80， all P<0.001）， along with significantly decreased numbers of adherent cells （F=73.65， 85.91， 138.00， all P<0.001）， while no significant differences were observed in cell cycle phase distribution in all three cell lines. ConclusionMAGED4 enhances proliferation and migration while suppressing adhesion in both HCC and normal hepatocytes， however， induces mesenchymal-like morphological changes exclusively in HCC cells， suggesting that MAGED4 is capable of driving HCC progression and may serve as a potential therapeutic target for HCC.

Objective: To investigate the predictive value of preoperative Neutrophil-to-Lymphocyte Ratio (NLR), serum albumin (ALB), and their combination for perioperative RBC transfusion in patients with hepatocellular carcinoma. Methods: A total of 1 027 patients clinically diagnosed with hepatocellular carcinoma who underwent radical surgery were retrospectively analyzed. The patients were divided into transfusion group and non-transfusion group according to whether they received red blood cell suspension transfusion. The perioperative clinical data of patients were compared between the groups. The predictive value of NLR combined with ALB for perioperative blood transfusion was analyzed by receiver operating characteristic (ROC) curve, and the optimal cutoff value was determined. Results: A total of 207 patients (20.16%, 207/1 027) received RBC transfusion. NLR combined with serum ALB concentration (ROC=0.712; P<0.001) was more accurate in predicting transfusion demand than NLR or ALB alone. The transfusion rate in the group with NLR>2.12 and ALB≤40.5 (45.32%, 92/203) was significantly higher than that in the group with NLR≤2.12 and ALB>40.5 (13.01%, 51/392) and the remaining combination group (14.81%, 64/432). Correlation analysis showed that NLR was statistically significantly correlated with ASA score, vascular invasion, satellite data, and the extent of liver resection. ALB was significantly correlated with ASA score, TNM stage, vascular invasion, satellite, and extent of hepatectomy. Conclusion: This study demonstrates that the combination of preoperative inflammatory factor NLR and serum ALB can effectively predict the perioperative blood transfusion needs of patients with hepatocellular carcinoma.

This article reports the diagnosis and management of a patient with maturity-onset diabetes of the young(MODY) type 10 combined with familial hypocalciuric hypercalcemia(FHH) type 1. The patient was a young female with a normal body mass index, who presented with hyperglycemia, hypercalcemia, and hypocalciuria. Diabetes-related autoantibodies were negative, the C-peptide level was normal, the parathyroid hormone level was not suppressed, and there was a three-generation family history of diabetes. Genetic testing confirmed the diagnosis of MODY10 combined with FHH1. This represents the first reported case and aims to offer insights for the early diagnosis and recognition of these two rare diseases, MODY and FHH.

Pathological myopic choroidal neovascularization(PM-CNV)is a common complication leading to vision loss in patients with pathological myopia.Its pathogenesis involves various factors,including mechanical traction,hypoxia,and inflammation.Anti-vascular endothelial growth factor therapy has been proven to be effective in the treatment of PM-CNV.Clinical trials have demonstrated that anti-vascular endothelial growth factor drugs,such as ranibizumab,aflibercept,and conbercept,can improve the vision of patients with PM-CNV,each with its characteristics and applicable scope.This article reviews the latest advancements in drug treatment for PM-CNV,aiming to provide valuable reference for the clinical management of this disease.

ObjectiveTo investigate the expression level of HBV cccDNA in patients in the convalescence stage of hepatitis B virus-related acute-on-chronic liver failure （HBV-ACLF） and its correlation with HBV markers and liver histopathological changes. MethodsA total of 30 patients in the convalescence stage of HBV-ACL who were hospitalized in The Ninth Hospital of Nanchang from January 2015 to October 2023 were enrolled as liver failure group， and 9 patients with chronic hepatitis B （CHB）， matched for sex and age， were enrolled as control group. The content of HBV cccDNA in liver tissue was measured， and its correlation with clinical data and laboratory markers was analyzed. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and a one-way analysis of variance or the Kruskal-Wallis H test was used for comparison between multiple groups； the Fisher’s exact test was used for comparison of categorical data between groups. A Spearman correlation analysis was performed. ResultsThe liver failure group had a significantly lower content of HBV cccDNA in liver tissue than the control group （-0.92±0.70 log₁₀ copies/cell vs -0.13±0.91 log₁₀ copies/cell， t=2.761， P=0.009）. In the liver failure group， there was no significant difference in the content of HBV cccDNA in liver tissue between the HBeAg-positive patients and the HBeAg-negative patients （P>0.05）； there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with different grades （G0-G2， G3， and G4） of liver inflammatory activity （P>0.05）； there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with different stages （S0-S2， S3， and S4） of liver fibrosis （P>0.05）； there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with negative HBV DNA and those with positive HBV DNA （P>0.05）. For the liver failure group， the content of HBV cccDNA in liver tissue was positively correlated with the content of HBV DNA in liver tissue （r=0.426， P=0.043） and was not significantly correlated with the content of HBV DNA in serum （P>0.05）. ConclusionThere is a significant reduction in the content of HBV cccDNA in liver tissue in the convalescence stage of HBV-ACLF. HBV cccDNA exists continuously and stably in liver tissue and can better reflect the persistent infection and replication of HBV than HBV DNA in serum and liver tissue.

Changes in hair follicle stem cells (HFSCs) can affect scalp aging and hair growth. With increasing age, HFSCs exhibit a decrease in quiescence maintenance and self-renewal capacity, as well as differentiation potential, leading to shortened hair growth cycles and even hair loss. This review summarizes recent research advances in the multifactorial interactions underlying hair loss, including the regulatory mechanisms of HFSC quiescence, the impact of aging on HFSC function, and aging of the stem cell microenvironment. Additionally, this review discusses the relationship between stem cells and hair shafts, and the mechanisms of action of stem cells in scalp aging, including alterations in signaling pathways, chromatin remodeling, and epigenetic regulation, etc. Furthermore, stem cell-based therapeutic strategies are summarized, such as the use of stem cells or their secreting exosomes, modulation of the stem cell microenvironment, and pharmacological interventions.

This article reports the diagnosis and management of a patient with maturity-onset diabetes of the young(MODY) type 10 combined with familial hypocalciuric hypercalcemia(FHH) type 1. The patient was a young female with a normal body mass index, who presented with hyperglycemia, hypercalcemia, and hypocalciuria. Diabetes-related autoantibodies were negative, the C-peptide level was normal, the parathyroid hormone level was not suppressed, and there was a three-generation family history of diabetes. Genetic testing confirmed the diagnosis of MODY10 combined with FHH1. This represents the first reported case and aims to offer insights for the early diagnosis and recognition of these two rare diseases, MODY and FHH.

Background Air pollution exposure has a significant impact on maternal and child health. However, the research on the association between ambient ozone (O₃) exposure during pregnancy and the risk of premature birth in newborns is limited, and the conclusions are inconsistent. Objective To investigate the association of ambient O₃ exposure during pregnancy with the risk of preterm birth in Guangdong Province. Methods Data of pregnant women in Guangzhou from 2013 to 2019 and Foshan from 2018 to 2023 were collected, and O₃ concentrations during different trimesters were assessed according to maternal residential addresses. Bilinear interpolation was used to evaluate the concentrations of air pollution. A cohort study design was adopted in our study. Restricted cubic spline curves were used to evaluate the exposure-response relationship between O₃ exposure and preterm birth risk and explore potential exposure threshold of O₃. Logistic regression models were used to evaluate the association of O₃ exposure with preterm birth. Results A total of 702 924 pregnant women were included in this study, of whom 43 051 (6.12%) were preterm. The average O₃ exposure concentrations of pregnant women during the first, second, third, and whole trimesters were 95.51, 97.51, 100.60, and 97.87 μg·m⁻³, respectively. We observed J-shaped associations between O₃ exposure and preterm birth risk during the second, third, and whole trimesters of pregnancy using restricted cubic spline curves. This study found that there were threshold concentrations between O₃ exposure and preterm birth risk during different gestational periods, and the threshold concentrations in the first, second, third, and whole trimesters were 112.32, 99.83, 111.74, and 112.46 μg·m⁻³, respectively. During the second, third, and whole trimesters of pregnancy, after adjusting for maternal age, baby sex, pre-pregnancy body mass index, mode of delivery, baby birth weight, gestational diabetes, and gestational hypertension, the odds ratios (OR) of preterm birth were 1.02 (95%CI: 1.01, 1.04), 1.02 (95%CI: 1.00, 1.03), and 1.17 (95%CI: 1.13, 1.21) for each 10 μg·m⁻³ increase in O₃ concentration above the O₃ threshold. No significant association was found between O₃ exposure and the risk of preterm birth during the first trimester. Conclusion There is a nonlinear association between the risk of preterm birth and O₃ exposure during pregnancy, and higher concentrations of O₃ exposure during pregnancy are associated with the risk of preterm birth. Above the O₃ threshold concentration during pregnancy, especially during the second, third, and whole trimesters, the risk of preterm birth elevates with the increase of O₃ exposure concentrations.

The aim of this study is to identify the linear Fc-binding epitope for IgG1 on bovine IgG Fc receptor Ⅱ(boFcγRⅡ)to understand the molecular basis of IgG-Fcγ interaction.The boFcγRⅡ molecules were expressed on cell surface of the boFcγR Ⅱ-transfected COS-7 cells.The extracel-lular domain of boFcγRⅡ was expressed in NS0 cells,and the boFcγRⅡ recombinant protein was purified from ascites by Ni-chelation chromatography.Peptides derived from the membrane-distal extracellular domain(EC2)of boFcγR Ⅱ were synthesized and conjugated to a carrier protein of IgG-free bovine serum albumin(BSA).Binding of bovine IgG1 to the different peptides was tested by dot-blot assay,and the IgG-binding peptide was further modified by truncation and mutation to identify the Fc-binding epitope as well as its key amino acids for Fc-binding.The inhibition effect of the Fc-binding peptide was determined by competitive ELISA and Fc-rosetting inhibition assay,re-spectively.The results showed that boFcγR Ⅱ molecules were stably expressed on surface of the transfected COS-7 cells,which showed about 90％rosetting with IgG1-RBCs.The soluble boFcγRⅡ recombinant protein specifically bound to bovine IgG1.The minimal effective peptide of 122FYQDRKSKIF131 of boFcγRⅡ was able to bind bovine IgG1 specifically,suggesting it repre-sents a linear Fc-binding epitope located in the putative C-C'loop of the EC2 domain on the recep-tor.The Ala-substitution of Phe122,Tyr123,Arg126,Lys127,Ser128,Lys129 or Phe131 within the linear epitope led to a complete loss of its IgG1-binding capability,indicating those residues are critical for IgG1-binding on boFcγRⅡ.The Fc-binding peptide inhibited bovine IgG1 binding to the soluble recombinant protein of boFcγRⅡ with IC50 of 20.05 μmol/L,and inhibited the rosette formation of bovine IgG1-sensitized RBCs on the boFcγRⅡ transfected cells with IC50 of 80.15 μmol/L.The re-sults indicate that boFcγRⅡ possesses the linear epitope for Fc-binding,and the Fc-binding pep-tide showed well capability of regulating boFcγR Ⅱ-IgG1 interaction on cell surface,thereby provi-ding a research foundation for understanding the IgG-Fcγ interaction.