OBJECTIVE: To explore the genetic variants and phenotypic characteristics of patients with Neurofibromatosis type I (NF1). METHODS: Twenty two NF1 patients who presented at Enze Medical (Center) Group in Taizhou between 2018 and 2024 were selected as the study subjects. Clinical phenotype and family history were collected for the patients. Whole exome sequencing (WES) was carried out for the 22 probands to screen the variants of NF1 gene. Candidate variants were verified by Sanger sequencing of their family members. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: K20230902). RESULTS: The 22 probands were diagnosed between the age of 5 months to 47 years old, and have all shown cafe au lait spots on their skin. Seventeen patients exhibited the phenotype at birth, and 11 had various degrees of neurofibromatosis. Among them, probands 1 and 13 underwent surgical resection of the tumor but had recurred, while proband 12 had amputation due to the huge size and serious impact of the neurofibroma and had no recurrence. Five patients had various degrees of scoliosis. In total 22 germline mutations and one somatic mutation were identified among the 22 families, with 5 variants unreported previously, including 1 nonsense mutation c.1603C>T (Q535*), 3 frameshift mutations [c.7268_7269delCA (Thr2423fs), c.2293del (Arg765Alafs*26), and c.5433_5438delinsGC (Phe1812ArgfsTer50)], and 1 deletion involving exons 41-44 of the NF1 gene and adjacent introns. Proband 13 was found to harbor germline mutation c.6796C>T (Gln2266Ter) and somatic mutation c.1019_1020del (Ser340Cysfs Ter12) in the peripheral blood and tumor tissue, respectively. Among the 22 NF1 probands, 6 had received treatment due to severe illness. Proband 1 had tumor resection in the right upper limb, but was found to have malignant lung tumor and died during follow-up. Proband 12 had multiple recurrence of neurofibroma in the left ring finger. Proband 4 underwent spinal correction surgery due to severe scoliosis. Proband 11 had died due to a central nervous system disease. Among the 22 germline mutations, 6 had led to the occurrence of truncated proteins, which may have a more severe impact on the phenotype. CONCLUSION This study investigated the genetic variants and clinical phenotypes of 22 NF1 families and identified 5 novel variants of the NF1 gene, which has expanded the genotypic and phenotypic spectra of the NF1. Preliminary studies have identified an association between truncated mutations, young age, and severe phenotypes, which may provide important clues for prognosis evaluation. For the clinical diagnosis and treatment of NF1, it is necessary to consider the phenotypic characteristics and genetic testing in combination with genetic counseling and long-term follow-up.
Humans ; Neurofibromatosis 1/pathology* ; Male ; Female ; Pedigree ; Adult ; Child ; Child, Preschool ; Middle Aged ; Adolescent ; Infant ; Young Adult ; Neurofibromin 1/genetics* ; Phenotype ; Asian People/genetics* ; Mutation ; Exome Sequencing ; East Asian People

AIM:This study aims to design pH/near-infrared(NIR)dual-responsive metal-organic framework composite nanoparticles co-loaded with indocyanine green(ICG)and sphingosine kinase 1(SphK1)siRNA(siSphK1),and to evaluate their anticancer efficacy against non-small-cell lung cancer A549 cells.METHODS:The ZIF-8 nanopar-ticles were synthesized and loaded with ICG and siSphK1 to prepare ZIF-8@ICG@siSphK1 nanoparticles.Their morpholo-gy,particle size,surface charge,and crystalline structure were characterized through transmission electron microscopy,dynamic light scattering,and X-ray diffraction.Stability,siSphK1 encapsulation and protection,and pH/NIR response were assessed.The gene silencing efficacy and anticancer activity in A549 cells were evaluated using Western blot,RT-qPCR,MTT assay,flow cytometry,and reactive oxygen species(ROS)fluorescence staining.RESULTS:The ZIF-8@ICG@siSphK1 nanoparticles exhibited a typical polyhedral structure with an average particle size of(76.8±0.9)nm and a ζ potential of(9.2±0.1)mV.The nanoparticles effectively encapsulated siSphK1,protecting it from RNase degra-dation,and demonstrated excellent NIR responsiveness with a photothermal conversion efficiency of 39.7%.After 10 h of 808 nm laser irradiation,siRNA cumulative release was significantly higher at pH 5.5 compared with pH 7.4.In A549 cells,the nanoparticles efficiently delivered siSphK1 under NIR irradiation,significantly down-regulated SphK1 gene ex-pression,inhibited cell proliferation,induced apoptosis,and increased intracellular ROS levels.CONCLUSION:The ZIF-8@ICG@siSphK1 nanoparticles effectively induce cytotoxic effects against A549 cells through gene silencing and pho-tothermal therapy.

Objective:To investigate the magnetic resonance imaging (MRI) features of pediatric and adolescent mediastinal T-lymphoblastic lymphoma (T-LBL) and to evaluate their predictive value for early treatment response.Methods:A retrospective, multicenter case series study was conducted on 49 pediatric and adolescent patients diagnosed with mediastinal T-LBL between September 2020 and May 2024 at the Fifth Affiliated Hospital of Zhengzhou University, Beijing Gaobo Boren Hospital, and Henan Cancer Hospital.All patients underwent chest MRI, including conventional MRI sequences and diffusion-weighted imaging.Tumor imaging characteristics were analyzed, and quantitative parameters such as minimum apparent diffusion coefficient (ADCmin), maximum ADC (ADCmax), and mean ADC (ADCmean) were measured.Treatment response was evaluated 15 days post-treatment.The patients were divided into a response group (complete or partial response, 26 cases) and a non-response group (progressive disease or minor response, 23 cases).The relationship between MRI features and treatment response was analyzed.The intraclass correlation coefficient was used to assess inter-reader agreement, and independent sample t-tests and chi-square tests were employed to compare differences between groups.Receiver operating characteristic (ROC) curve analysis was conducted to evaluate the predictive performance of imaging parameters. Results:Significant differences were observed between the response and non-response groups in ADC values [ADCmin (0.80±0.41)×10 -3 mm 2/s vs.(1.23±0.70)×10 -3 mm 2/s, ADCmax (1.14±0.48)×10 -3 mm 2/s vs.(1.92±0.77)×10 -3 mm 2/s, ADCmean (0.98±0.42)×10 -3 mm 2/s vs.(1.56±0.74)×10 -3 mm 2/s] and the maximum tumor diameter was [(11.92±3.61) cm vs.(8.17±2.46) cm] (all P<0.05).ROC curve analysis showed that ADCmax had the highest predictive efficiency for treatment response, with an area under the curve (AUC) of 0.853 (95% CI: 0.790-0.910), sensitivity of 92.3%, and specificity of 65.2%.The AUC for the maximum tumor diameter was 0.814, demonstrating its excellent predictive performance. Conclusions:MRI features, particularly ADC values and the maximum tumor diameter, can effectively predict treatment response in pediatric and adolescent mediastinal T-LBL.

Objective:To observe and preliminarily analyze the risk factors related to silicone oil dependence after pars plana vitrectomy (PPV) in open globe injuries (OGI) .Methods:A retrospective clinical study. This study included 211 patients with OGI who received PPV treatment from January 1, 2005 to December 31, 2016 in Eye Injury Vitrectomy Study database. Silicone oil dependence was defined as the intraocular pressure <10 mm Hg (1 mm Hg= 0.133 kPa) in silicone oil tamponade eye 6 months after PPV. The patient's age, intraocular pressure, type of injury, length of eyeball wound, as well as the presence of ciliary body injury, choroidal injury, and retinal resection and/or defect area were recorded in detail. Logistic multifactorial regression analyzed the correlation between age, eye wound length and ciliary body iinjury, choroidal injury and retinal resection and/or defect severity and silicone oil dependence.Results:211 OGI eyes underwent vitrectomy were included, the mean age of the study population was 32.93 years (range 1-73 years), and the median follow-up period was 20.06 months (range 5.85-90.58 months). Among the 211 eyes, 121 (57.3%, 121/211), 52 (24.6%, 52/211), 23 (10.9%, 23/211) and 15 (7.1%, 15/211) were rupture, perforation, intraocular foreign bodies and penetrating injury, respectively. 98 eyes (46.4%, 98/211) of silicone oil dependence in total, among them, 64 (65.3%, 64/98), 19 (19.4%, 19/98), 9 (9.2%, 9/98) and 6 (6.1%, 6/98) eyes were rupture, perforation, intraocular foreign body and penetrating injury, respectively. Correlation analysis revealed that silicone oil dependence was significantly associated with ciliary body injury [odds ratio ( OR) = 2.150, 95% confidence interval ( CI) 1.052-4.393], choroidal injury ( OR=3.233, 95% CI 1.454-7.191), and retinal injury ( OR=2.731, 95% CI 1.393-5.353). No correlation was found with age ( OR= 0.981, 95% CI 0.960-1.002) or ocular wound length ( OR=1.716, 95% CI 0.987-2.984) ( P> 0.05). Conclusion:Silicone oil dependency following PPV in OGIs are associated with ciliary body, choroidal, and retinal injuries, but no association with age or length of the ocular wound.

OBJECTIVE: To determine the types of genetic variants in six Chinese pedigrees affected with Marfan syndrome (MFS) and analyze their clinical characteristics and molecular pathogenesis. METHODS: Six MFS pedigrees presented at the Taizhou Enze Medical Center (Group) between 2017 and 2022 were selected as the study subjects. Clinical data of pedigrees were retrospectively analyzed. Peripheral blood samples were collected from the probands and their family members for the extraction of genomic DNA. Whole exome sequencing (WES) was carried out. Candidate variants of the FBN1 gene were verified by Sanger sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), pathogenicity of the candidate variants was assessed. AlphaFold3 and PyMOL software were used for homology modeling of the FBN1 protein and analysis of its three-dimensional structure and amino acid sequence conservation. This study was approved by the Medical Ethics Committee of Taizhou Enze Medical Center (Group) (Ethics No. 20231002). RESULTS: Cardiovascular system abnormalities were noted in all pedigrees, ocular abnormalities were present in pedigrees 2 and 5, skeletal system abnormalities were presented in pedigrees 1, and 4 to 6. FBN1 gene mutations were identified in all pedigrees, including c.1957_1958dupGT (p.Asp654fs), c.5014T>A (p.Cys1672Ser), c.8135delC (p.Pro2712fs), c.2302G>T (p.Glu768*), c.3473A>G (p.Glu1158Gly) and c.6169C>T (p.Arg2057*), with each involving a different exon. Four variants were rated as pathogenic, one as likely pathogenic, and one as variant of uncertain significance. Among these, c.5014T>A (p.Cys1672Ser), c.1957_1958dupGT (p.Asp654fs), c.8135delC (p.Pro2712fs), and c.2302G>T (p.Glu768*) were unreported previously. Bioinformatic analysis with SIFT and PolyPhen-2 predicted that the c.5014T>A (p.Cys1672Ser) and c.3473A>G (p.Glu1158Gly) variants were deleterious. Protein homologous sequence alignment analysis revealed that the four novel mutation sites are highly conserved across various species. Homology modeling of the FBN1 protein three-dimensional structure indicated that the six variant sites in the amino acid sequence are all close to hydrogen bonds and may alter the secondary and tertiary structures to varying degrees, thereby confirmed the relationship between the variants and MFS. CONCLUSION Four novel variants of the FBN1 gene have been discovered in this study, which has enriched the mutational and phenotypic spectrum of MFS and provided a basis for disease diagnosis and genetic counseling.
Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Middle Aged ; Young Adult ; China ; East Asian People/genetics* ; Exome Sequencing ; Fibrillin-1/genetics* ; Marfan Syndrome/genetics* ; Mutation ; Pedigree ; Retrospective Studies ; Adipokines

Objective:To determine the types of genetic variants in six Chinese pedigrees affected with Marfan syndrome (MFS) and analyze their clinical characteristics and molecular pathogenesis.Methods:Six MFS pedigrees presented at the Taizhou Enze Medical Center (Group) between 2017 and 2022 were selected as the study subjects. Clinical data of pedigrees were retrospectively analyzed. Peripheral blood samples were collected from the probands and their family members for the extraction of genomic DNA. Whole exome sequencing (WES) was carried out. Candidate variants of the FBN1 gene were verified by Sanger sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), pathogenicity of the candidate variants was assessed. AlphaFold3 and PyMOL software were used for homology modeling of the FBN1 protein and analysis of its three-dimensional structure and amino acid sequence conservation. This study was approved by the Medical Ethics Committee of Taizhou Enze Medical Center (Group) (Ethics No. 20231002). Results:Cardiovascular system abnormalities were noted in all pedigrees, ocular abnormalities were present in pedigrees 2 and 5, skeletal system abnormalities were presented in pedigrees 1, and 4 to 6. FBN1 gene mutations were identified in all pedigrees, including c. 1957_1958dupGT (p.Asp654fs), c. 5014T>A (p.Cys1672Ser), c. 8135delC (p.Pro2712fs), c. 2302G>T (p.Glu768*), c. 3473A>G (p.Glu1158Gly) and c. 6169C>T (p.Arg2057*), with each involving a different exon. Four variants were rated as pathogenic, one as likely pathogenic, and one as variant of uncertain significance. Among these, c. 5014T>A (p.Cys1672Ser), c. 1957_1958dupGT (p.Asp654fs), c. 8135delC (p.Pro2712fs), and c. 2302G>T (p.Glu768*) were unreported previously. Bioinformatic analysis with SIFT and PolyPhen-2 predicted that the c. 5014T>A (p.Cys1672Ser) and c. 3473A>G (p.Glu1158Gly) variants were deleterious. Protein homologous sequence alignment analysis revealed that the four novel mutation sites are highly conserved across various species. Homology modeling of the FBN1 protein three-dimensional structure indicated that the six variant sites in the amino acid sequence are all close to hydrogen bonds and may alter the secondary and tertiary structures to varying degrees, thereby confirmed the relationship between the variants and MFS. Conclusion:Four novel variants of the FBN1 gene have been discovered in this study, which has enriched the mutational and phenotypic spectrum of MFS and provided a basis for disease diagnosis and genetic counseling.

AIM:This study aims to design pH/near-infrared(NIR)dual-responsive metal-organic framework composite nanoparticles co-loaded with indocyanine green(ICG)and sphingosine kinase 1(SphK1)siRNA(siSphK1),and to evaluate their anticancer efficacy against non-small-cell lung cancer A549 cells.METHODS:The ZIF-8 nanopar-ticles were synthesized and loaded with ICG and siSphK1 to prepare ZIF-8@ICG@siSphK1 nanoparticles.Their morpholo-gy,particle size,surface charge,and crystalline structure were characterized through transmission electron microscopy,dynamic light scattering,and X-ray diffraction.Stability,siSphK1 encapsulation and protection,and pH/NIR response were assessed.The gene silencing efficacy and anticancer activity in A549 cells were evaluated using Western blot,RT-qPCR,MTT assay,flow cytometry,and reactive oxygen species(ROS)fluorescence staining.RESULTS:The ZIF-8@ICG@siSphK1 nanoparticles exhibited a typical polyhedral structure with an average particle size of(76.8±0.9)nm and a ζ potential of(9.2±0.1)mV.The nanoparticles effectively encapsulated siSphK1,protecting it from RNase degra-dation,and demonstrated excellent NIR responsiveness with a photothermal conversion efficiency of 39.7%.After 10 h of 808 nm laser irradiation,siRNA cumulative release was significantly higher at pH 5.5 compared with pH 7.4.In A549 cells,the nanoparticles efficiently delivered siSphK1 under NIR irradiation,significantly down-regulated SphK1 gene ex-pression,inhibited cell proliferation,induced apoptosis,and increased intracellular ROS levels.CONCLUSION:The ZIF-8@ICG@siSphK1 nanoparticles effectively induce cytotoxic effects against A549 cells through gene silencing and pho-tothermal therapy.

This article reports the diagnosis and treatment of a patient with diffuse large B-cell lymphoma (DLBCL) initially manifested as acquired von Willebrand syndrome (AvWS), along with a literature review. The patient, a 22-year-old male, was diagnosed with hereditary von Willebrand disease at the initial stage due to repeated mucosal bleeding, and was later diagnosed with DLBCL (non germinal center type, Ann Arbor stage Ⅳ) due to chest wall mass. Through chemotherapy combined with autologous hematopoietic stem cell transplantation and zebutinib maintenance therapy, the patient achieved sustained complete remission, and the coagulation function returned to normal. This case provides an important reference for the diagnosis and treatment of lymphoma associated AvWS, and highlights the importance of early recognition of basic diseases.

To analyze the current state and issues of ideological education for graduate students of general practice, from April 10 to Dec 19, 2024, a literature review was conducted on publications from Jan 1, 2001, to Dec 19, 2024, in the CNKI, Wanfang, and VIP databases to identify research entry points. Purposive sampling was used to select postgraduate students majoring in general practice at Guangzhou Medical University and their corresponding faculty as the study participants. A semi-structured in-depth interview method was employed to investigate the current state of ideological and political education. Faculty evaluations of students and their recommendations for ideological and political education were also considered. Thematic framework analysis was applied to the interview data for categorization, integration, and theme extraction. The literature review revealed that enhancing professional identity can strengthen career commitment, leading to the design of an ideological and political education interview outline focused on professional belief and professional identity. The study included 13 general practice postgraduates (mean age: 24.5±1.4 years), of whom 8 were directional students and 5 were non-directional students. Additionally, 7 general practice faculty members were included (mean age: 41.3±5.4 years), with an average of 16.3±5.6 years of experience in general practice education. Five themes were extracted from the interviews: the necessity of ideological and political education, the relationship between professional belief and its educational effects, the intrinsic motivation of ideological beliefs, the influence of external factors on beliefs, and effective approaches to ideological and political education. The study also identified current issues in ideological and political education for general practice postgraduates, including inadequate career planning guidance, reluctance to work in underdeveloped areas, over-reliance on the role modeling of faculty mentors, lack of systematic courses and frameworks for ideological and political education, and limited opportunities for team projects and communication platforms. The current state of ideological and political education for general practice postgraduates reveals deficiencies in career planning support, systematic course construction, practical opportunities, and communication platforms. Education methods remain overly simplistic and are significantly influenced by external factors, underscoring the need for optimization and enhancement.

Objective:To investigate the correlations between physical, psychological and social frailty in elderly patients with multimorbidity.Methods:This study utilized a mixed method. A questionnaire survey was conducted from February to June 2024, among elderly patients with multimorbidity attending 4 primary health care centers in urban Beijing selected by the convenience sampling method. The FRAIL Frailty Assessment Scale, WHO-5 Index of Well-Being Scale, and HALFT Scale were used to assess the patients′ physical, psychological, and social frailty, respectively. Spearman correlation analysis was used to analyze the correlation between different dimensions of frailty in elderly with multimorbidity. Logistic regression model was used to analyze the factors influencing physical, psychological and social frailty. The elderly with multimorbidity who were assessed to have at least 1 or more types of frailty in the quantitative study were selected for in-depth interviews in the form of online and offline combination. The topics of in-depth interview included the real experience of the different dimensions of frailty, the possible causes and the difficulties caused. The sample size was determined according to the principle of information saturation. Thematic analysis was used to summarize, code and analyze the interview data.Results:A total of 919 participants were included in the quantitative study, with a mean age of (74.09±6.03) years, 329(35.80%) were males and 590(64.20%) were females. The prevalence of physical, psychological, and social frailty was 17.85%(164/919), 21.44%(197/919), 11.21%(103/919), respectively. A total of 21 participants were included in the qualitative study, with a mean age (76.90±5.13)years, 5(23.81%) males and 16(76.19%) females. Spearman correlation analysis showed that physical and psychological frailty were moderately correlated ( r=0.311, P<0.001), psychological and social frailty were weakly correlated ( r=0.218, P<0.001), and physical and social frailty were weakly correlated ( r=0.267, P<0.001). Logistic regression analysis showed that the age, the number of multimorbidities, the psychological frailty and social frailty were the influencing factors for physical frailty (all P<0.05). The gender, number of multimorbidity, type of medication taken, physical frailty and social frailty were influencing factors of psychological frailty (all P<0.05). And age, number of multimorbidities, physical frailty and psychological frailty were influencing factors of social frailty (all P<0.05). A total of 3 themes were extracted through in-depth interviews, namely, "physical and psychological frailty are interrelated""physical and social frailty are interrelated", and "psychological and social frailty are interrelated". Conclusions:The physical, psychological, and social frailty in elderly patients with multimorbidity interacts with each other. Whereas the number of multimorbidities is a common risk factor for all three.