Background Carbendazim (CBZ), a widely used benzimidazole fungicide, has raised increasing concerns regarding the health risks associated with its residues. However, the toxic effects and associated mechanisms of CBZ on the skeletal system have not been reported. Objective To elucidate the effects of carbendazim on osteogenic differentiation and its underlying mechanisms. Methods MC3T3-E1 mouse pre-osteoblastic cells were treated with 1, 10, and 100 μmol·L⁻¹ CBZ for 24 h to examine cell viability, alkaline phosphatase (ALP) activity, bone nodule formation, reactive oxygen species (ROS) level, malondialdehyde (MDA) content, and nitric oxide synthase (NOS) activity. Transcriptomics was used to identify differentially expressed genes (DEGs) in osteoblasts exposed to CBZ. Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene set enrichment analysis (GSEA) were employed to analyze the potential biological pathways of DEGs. Real-time polymerase chain reaction (RT-PCR) and Western blot were used to validate changes in gene and protein expression. Results Exposure to 10 and 100 μmol·L⁻¹ CBZ significantly reduced osteoblast viability, ALP activity, bone nodule formation, and NOS activity, while increasing intracellular ROS levels. CBZ at 100 μmol·L⁻¹ concentration significantly elevated MDA level (P < 0.05). The transcriptomic analysis revealed that 1 μmol·L⁻¹ CBZ treatment resulted in 385 significantly DEGs. The KEGG enrichment analysis revealed that CBZ significantly affects hormone regulation pathways (including parathyroid hormone, growth hormone, dopamine, and oxytocin), mitogen-activated protein kinase (MAPK) and cyclic GMP-dependent protein kinase G (cGMP-PKG) signaling pathways, focal adhesion and adherens junction, as well as the NOD-like receptor signaling pathway and the mRNA surveillance (NMD) pathway. The results of GSEA showed that CBZ significantly inhibited the bile acid metabolism and the Wnt/β-catenin pathway in osteoblasts. The validation results demonstrated that CBZ significantly suppressed the mRNA expression of Wnt3a and β-catenin, as well as the protein expression of Runx2 and Osterix in the Wnt/β-catenin pathway. Conclusion CBZ exposure exhibits potential skeletal toxicity, and its mechanism is through promoting oxidative stress, interfering with the Wnt/β-catenin pathway in osteogenic differentiation, thereby inhibiting the bone formation function of osteoblasts.

OBJECTIVE: To summarize the clinical and genetic characteristics of children with Silver-Russell syndrome (SRS) and improve the recognition of this disease. METHODS: A retrospective analysis was conducted on the clinical manifestations and genetic testing results of 29 children with SRS diagnosed at the Children's Hospital Affiliated to Zhengzhou University between March 2016 and June 2025. RESULTS: The 29 children had included 18 boys and 11 girls, with the age ranging from 2 months to 16 years. Their primary clinical manifestations included postnatal growth retardation (100%), small for gestational age (SGA) (100%), characteristic facial features (90%), limb asymmetry (83%), feeding difficulties (76%), ulnar deviation of the fifth finger (69%), body mass index (BMI) of < -2 SD (62%), and abnormal bone age (55%), including 15 cases with delayed bone age for an average of 1.5 years and 1 case with advanced bone age for 2.5 years. Additional manifestations included abnormal sexual development in 11 cases (38%), dental malocclusion in 11 cases (38%), allergic diseases in 10 cases (34%), cardiac diseases in 9 cases (31%), skeletal abnormalities in 7 cases (24%), renal hypoplasia in 5 cases (17%), and abnormal cranial MRI findings in 5 cases (17%). Twenty children were treated with recombinant human growth hormone (rhGH) at a dose of 0.1 ~ 0.15 U/(kg.d). Among them, 7 cases achieved annual height increase of ≥ 10 cm, 11 cases achieved annual height increase of ≥ 5 ~ 9 cm, and 2 cases achieved annual height increase < 5 cm. Twenty three children exhibited hypomethylation of imprinted genes in the chromosome region of 11p15, 4 presented maternal uniparental disomy of chromosome 7 [UPD(7)mat], and 2 had harbored nonsense variants of the HMGA2 gene. CONCLUSION SRS patients may present with diverse clinical manifestations including postnatal growth retardation, SGA, characteristic facial features, limb asymmetry, feeding difficulties, and ulnar deviation of the fifth finger. Most patients may exhibit abnormal methylation in the 11p15 region. rhGH therapy can improve the height of these patients.
Humans ; Silver-Russell Syndrome/diagnosis* ; Male ; Female ; Child ; Child, Preschool ; Infant ; Adolescent ; Retrospective Studies

ObjectiveTo explore the biological mechanism of bone loss caused by perfluorooctanoic acid (PFOA) through transcriptomic analysis, and to provide new insights into regulating perfluoroalkyl substances (PFAS) applications and the prevention of hazards affecting bone health. MethodsMouse embryonic osteoblast precursor cells (MC3T3-E1) were exposed to 0.1, 1, 10, and 100 μmol·L^-¹ PFOA for 24 hours to assess the effects on cell viability and alkaline phosphatase (ALP) activity, and to determine the critical concentration of PFOA toxicity. The transcriptome sequencing (RNA-seq) was performed to identify differentially expressed genes (DEGs) induced by PFOA. Gene ontology (GO) analysis and gene set enrichment analysis (GSEA) were conducted to identify significantly affected gene pathways. Additionally, Seahorse XF metabolic phenotyping and reverse transcription polymerase chain reaction (RT-PCR) were used to validate the key pathways. ResultsExposure to 10 and 100 μmol·L^-¹ PFOA significantly reduced the cell viability and ALP activity of MC3T3-E1 cells. Therefore, the results of transcriptomic analysis for 10 μmol‧L^-1 PFOA exposure found that a total of 80 DEGs were identified, including 32 upregulated genes and 48 downregulated genes. According to GO analysis, PFOA mainly affected cellular components such as mitochondrion and nucleus, molecular functions involving GTPase activity and GTP binding, as well as biological process related to mRNA processing. GSEA identified the downregulation of the β-oxidation of fatty acid pathway in mitochondria. Metabolic phenotyping reserches showed that PFOA indeed reduced mitochondrial aerobic respiration capacity and adenosine triphosphate (ATP) production, and the ratio of ATP production from cellular aerobic respiration to glycolysis was significantly decreased as well. The mRNA expression of glucose metabolism-related genes (GK, G6PD, and CS), as well as fatty acid metabolism-related genes (CPT1A and CPT2), were significantly downregulated. ConclusionPFOA reduces bone formation by inhibiting energy metabolism and β-oxidation of fatty acid pathways in osteoblasts, whihc lays the foundation for revealing the mechanism of PFOA exposure induced bone loss.

Doxorubicin (Dox) is an anthracycline drug widely applied in various malignancies. However, the fatal cardiotoxicity induced by Dox limits its clinical application. Post-transcriptional protein modification via ubiquitination/deubiquitination in cardiomyocytes mediates the pathophysiological process in Dox-induced cardiotoxicity (DIC). In this study, we aimed to clarify the regulatory role and mechanism of a deubiquitinating enzyme, ubiquitin-specific peptidase 13 (USP13), in DIC. RNA-seq analysis and experimental examinations identified that cardiomyocyte-derived USP13 positively correlated with DIC. Mice with cardiac-specific deletion of USP13 were subjected to Dox modeling. Adeno-associated virus serotype 9 (AAV9) carrying cTNT promoter was constructed to overexpress USP13 in mouse heart tissues. Cardiomyocyte-specific knockout of USP13 exacerbated DIC, while its overexpression mitigated DIC in mice. Mechanistically, USP13 deubiquitinates the stimulator of interferon genes (STING) and promotes the autolysosome-related degradation of STING, subsequently alleviating cardiomyocyte inflammation and death. Our study suggests that USP13 serves a cardioprotective role in DIC and indicates USP13 as a potential therapeutic target for DIC treatment.

ObjectiveTo explore the role of flurochloridone (FLC) on osteogenic differentiation and the potential mechanism of inhibiting bone formation, and to provide new insights into bone health risks associated with FLC pesticide exposure. MethodsNeonatal rat skull differentiation primary osteoblast model was used to investigate the effects of 1, 10 and 100 μmol·L^-1 FLC exposure on cell viability, osteogenic differentiation alkaline phosphatase (ALP) activity, and bone mineralization nodule formation, respectively. The potential mechanism underlying the inhibition of FLC on osteoblast differentiation was analyzed using osteogenic differentiation gene chip technique, and the expression of key genes and proteins in the pathway was validated using reverse transcription polymerase chain reaction (RT-PCR) and protein immunoblotting (Western blot) methods. ResultsExposure to FLC at a concentration of 100 μmol·L⁻¹ reduced cell proliferation, ALP activity, and the formation of mineralized nodules in primary osteoblasts. Gene chip analyses revealed that exposure to 10 μmol·L⁻¹ FLC induced 15 differentially expressed genes (DEGs). Among these, MMP9 and Tnf were up-regulated, while Nkx3⁃2, Tuft1, Bmp2, Col12a1, Pparg, Enam, Igf1, Bmp5, Bmp3, Calcr, Egf, Igfbp3, and Col14a1 were down-regulated. Results of protein-protein interaction analyses and gene ontology enrichment analyses indicated that FLC inhibited the BMP/SMAD pathway involved in osteogenic differentiation. FLC suppressed the protein expression of BMP2 and Osterix, as well as the expression of key genes critical for osteogenic differentiation and ossification, such as BMP2, Runx2, SMAD1, and SMAD5 in the BMP/SMAD pathway. ConclusionFLC affects osteogenic differentiation and bone formation potential by regulating the BMP/SMAD axis and the expression of osteogenic genes, suggesting its potential risk in bone metabolism.

OBJECTIVE: To explore the clinical features, genetic characteristics in a child with Miller-McKusick-Malvaux syndrome (3MS) type 1 caused by CUL7 gene variant. METHODS: A child diagnosed with 3MS type 1 at the Children's Hospital Affiliated to Zhengzhou University in February 2021 was selected as the subject of this study. Peripheral blood samples were collected from the child and her parents for genomic DNA extraction. Whole exome sequencing (WES) was performed on the child, and Sanger sequencing was used to validate the candidate variants and analyze their pathogenicity. A literature search was conducted using the keywords "3M syndrome" in the China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, and PubMed databases from inception to December 2024. The clinical data of Chinese children with 3MS reported in the literature were summarized. This study was approved by the Medical Ethics Committee of the Children's Hospital Affiliated to Zhengzhou University (Ethics No. 2024-K-020). RESULTS: The child was a 6-year-old and 2-month-old female with facial dysmorphism, skeletal abnormalities, and growth and developmental delay. WES revealed compound heterozygous variants in the CUL7 gene: c.2686G>T (p.E896*) and c.1200delT (p.R401Gfs66). Sanger sequencing confirmed that these two variants were inherited from the child's father and mother, respectively. According to the American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants, c.2686G>T (p.E896) was classified as a pathogenic (PVS1+PM2_Supporting+PM3), and c.1200delT (p.R401Gfs*66) was classified as a likely pathogenic (PVS1+PM2_Supporting). Based on the literature search strategy, 18 relevant articles were identified, including a total of 32 Chinese cases of 3MS, of which 8 were fetuses. A total of 32 Chinese 3MS cases were included in the literature review, of which 8 were fetuses. The majority of these cases carried variants in the CUL7 gene (20/32, 62.5%) and OBSL1 gene (12/32, 37.5%). The main clinical manifestations included intrauterine or postnatal growth and developmental delay (32/32, 100.0%), triangular facies (27/32, 84.3%), and skeletal abnormalities (21/32, 65.6%). CONCLUSION The compound heterozygous variants c.2686G>T (p.E896*) and c.1200delT (p.R401Gfs*66) in the CUL7 gene are likely the genetic cause of 3MS type 1 in the child. For children presenting with facial dysmorphism, skeletal abnormalities, and intrauterine or postnatal growth and developmental delay, 3MS should be considered as a differential diagnosis.
Humans ; Cullin Proteins/genetics* ; Female ; Child ; Limb Deformities, Congenital/genetics* ; Exome Sequencing ; Mutation ; Child, Preschool ; Dwarfism ; Muscle Hypotonia ; Spine/abnormalities*

Epidemiology and medical statistics are essential courses for undergraduate students majoring in clinical medicine. By studying the two courses, they can obtain the core skills for their future clinical practice. High-level medical schools both at home and abroad have accumulated successful experiences in curriculum, teaching methods and teaching models of the two disciplines. These colleges have also carried out the exploration of the curriculum reform centering on "organ systems integration". This paper summarizes the current status of epidemiology and medical statistics teaching and curriculum integration in representative medical schools both at home and abroad, and puts forward suggestions for deepening teaching reform and optimizing the curriculum system to provide reference for the integration of epidemiology and medical statistics curriculums for undergraduate students majoring in clinical medicine in China.

Objective To explore the application effect of upper abdominal moxibustion combined with bedside ultrasound monitoring of gastric residual volume(GRV)in pre-pyloric feeding in stroke patients.Methods Eighty stroke patients admitted to the Department of Rehabilitation Medicine of Zhejiang Provincial People's Hospital from January 1,to December 31,2023 were selected as the study subjects.They were divided into control group(n=38)and observation group(n=42)using a random number table method.All patients had a nasogastric tube for pre-pyloric feeding.The control group used the traditional syringe aspiration method to monitor GRV,while the observation group used upper abdominal moxibustion combined with bedside ultrasound to monitor GRV.The study compared the differences between two groups in terms of enteral nutrition intolerance,feeding complications,enteral nutrition compliance rate within 7 days of admission,time to achieve enteral nutrition compliance,and changes in hemoglobin(Hb),serum prealbumin,serum albumin(ALB),and serum transferrin before and after 14 days of feeding.Results The incidence rates of vomiting,abdominal distention,intra-abdominal hypertension,reflux,and aspiration pneumonia in observation group were lower than those in control group(P＜0.05).The rate of achieving intestinal nutrition standard within 7 days of hospitalization was significantly higher in observation group compared to the control group.The time to achieve intestinal nutrition standard was shorter in observation group compared to control group.Furthermore,after 14 days of feeding,the levels of Hb and ALB in observation group were higher than those in control group,and the differences were statistically significant(P＜0.05).Conclusion Upper abdominal moxibustion combined with bedside ultrasonic monitoring of GRV can significantly reduce intestinal nutrition intolerance and feeding complications during pre-pyloric feeding in stroke patients,shorten the time to achieve nutritional benchmarks,and improve nutritional status.

Per- and polyfluoroalkyl substances （PFASs） are a new type of persistent organic pollutants with global attention. They have shown multiple toxic effects due to their persistent accumulation in human body through exposure to environmental media such as drinking water， food， atmosphere， and soil. However， the bone toxicity of PFASs has not attracted enough attention. It is believed that the exposure and accumulation of PFASs in human have a significant impact on the bone health， especially hindering the healthy bone development in infants and adolescents， and aggravating the occurrence of bone loss and fracture in the elder populations. This paper will review the research progress of the effects of PFASs exposure on bone health indicators such as bone mineral density， and discuss the mechanisms of PFAS in bone toxicity. This review will provide references for revealing the effects of PFASs exposure on bone health and their toxic mechanisms.

ObjectiveTo establish an High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometry （HPLC-ICP-MS） method for determination of six arsenic species in human urine，including arseniccholine （AsC）， arsenobetaine （AsB）， arsenite （As³⁺）， dimethylarsinic acid （DMA⁵⁺）， monomethylarsonic acid （MMA⁵⁺）， and arsenate （As⁵⁺）. MethodsThe pH value of mobile phase and the content of anhydrous ethanol were optimized. Ammonium carbonate （50 mmol·L^-1， containing 2% anhydrous ethanol， pH-8.5） mobile phase was selected. Cl^- interference was eliminated by He mode. The arsenic species in 10-fold diluted human urine samples were separated by an Hamilton PRP X-100 anionic column. A method for the determination of six arsenic species was established. ResultsSix arsenic species could be separated in 13 minutes. The linear correlation coefficients were above 0.999. The limits of detection were 0.10‒0.20 μg·L^-1， and the limits of quantification were 0.30‒0.50 μg·L^-1. Precision experiments showed that RSD ranged from 5.96% to 9.07% when adding concentration 0.20 μg·L^-1； from 2.48% to 6.38% when adding concentration 2.00 μg·L^-1； and from 1.41% to 2.57% when adding concentration 5.00 μg·L^-1. Accuracy test showed that the recoveries were 80%‒125%. ConclusionThe established HPLC-ICP-MS method for determination of six arsenic species in human urine is rapid， accurate and sensitive. It can be applied to the determination of arsenic species in human urine.