Objective To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly(VM)in fetuses.Methods Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA,and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups.Three miRNAs with significant differential expression between the two groups,whose high expression was associated with VM,were selected for verification with RT-qPCR.Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes AKT3 and CCDC88C.Gene ontology(GO)and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.Results We identified a total of 272 differentially expressed miRNAs in VM cases,including 43 up-regulated and 229 down-regulated miRNAs.The target genes of these differential miRNAs were associated with DNA and transcription factor binding,transmembrane transporter and nucleic acid binding transcription factor activity,and cell developmental process.These miRNAs were mostly enriched in the MAPK,cGMP-PKG and Wnt signaling pathways.Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group(P<0.05),which was consistent with miRNA sequencing results;let-7b-5p expression level was significantly lower in VM group,which was contrary to miRNA sequencing result.Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating AKT3 or CCDC88C expressions.Conclusions The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK,PI3K-Akt,Wnt and cGMP-PKG signaling pathways.

Objective:To study the differential expression and correlation of salivary secretory immunoglobulin A (sIgA) and peripheral blood hemoglobin (Hb) levels in children with different severity of dental caries.Methods:This study was a case-control study, selecting 106 children with dental caries who received treatment at Jiaxing Maternal and Child Health Hospital from December 2020 to June 2022 as the observation group, and 100 healthy children without dental caries during the same period as the control group. We compared the salivary sIgA and peripheral blood Hb levels between two groups of children. Children with dental caries were divided into mild group (2-3 teeth), moderate group (4-7 teeth), and severe group (≥8 teeth) based on the number of missing teeth. The salivary sIgA and peripheral blood Hb levels of children with different degrees of dental caries were compared. Pearson correlation analysis was used to evaluate the correlation between the number of missing teeth and salivary sIgA and peripheral blood Hb levels. Multivariate logistic regression analysis was used to evaluate the influencing factors of dental caries in children.Results:The salivary sIgA and peripheral blood Hb levels in the observation group were lower than those in the control group, and the differences were statistically significant (all P<0.05). There were 28 cases in the mild group, 50 cases in the moderate group, and 28 cases in the severe group, respectively. There were statistically significant differences in salivary sIgA and peripheral blood Hb levels among children with different severity of dental caries. As the severity of the disease increased, salivary sIgA and peripheral blood Hb levels showed a decreasing trend (all P<0.05). Pearson correlation analysis showed that the number of missing teeth was negatively correlated with salivary sIgA and peripheral blood Hb levels ( r=-0.572, -0.584; P<0.001). Multivariate logistic regression analysis showed that salivary sIgA ( OR=0.660, 95% CI: 0.463-0.941) and peripheral blood Hb ( OR=0.680, 95% CI: 0.505-0.916) were protective factors for the occurrence of dental caries in children (all P<0.05). Conclusions:Salivary sIgA and peripheral blood Hb levels in children with dental caries are low and negatively correlated with the severity of the disease.

Objective To investigate the changes in G protein-coupled receptor 124 (Gpr124) expression in human retinal microvascular endothelial cells (HRMEC)under high-glucose condition and the regulatory effect of Gpr124 interference on HRMEC.Methods Immunofluorescence assay was used to observe the expression of Gpr124 in HRMEC.The cells were divided into a blank group and a high glucose group.CCK-8 assay and EdU cell proliferation assay were used to detect cell viability and proliferation,respectively.The Gpr124 expression was knocked down by transducting lentiviral virus carrying shRNA targeting Gpr124.Thus,the cells were divided into a blank control group,a high glucose control group,a high glucose+shRNA group,and a high glucose+Gpr124 shRNA group.Cell scratch test was performed to evaluate cell migration,and Matrigel plug assay was employed to assess the cell tubule formation.Western blotting was applied to detect the protein levels of Gpr124,VEGFA,MMP9,and β-catenin.Results The HRMEC from the high glucose group showed enhanced cell viability and increased number of proliferating cells compared to the cells of the blank group (P<0.01).High glucose treatment also induced increased expression levels of Gpr124,VEGFA and MMP9 (P<0.01),and larger migratory area and in vitro angiogenic area as well as longer tube diameter at all time points when compared with the conditions in the blank control group (P<0.01).However,knockdown of Gpr124 resulted in a significant decrease in HRMEC migration and in vitro angiogenic capacity,accompanied by decreases in protein expression of VEGFA,MMP9 and β-catenin (P<0.01).Conclusion Gpr124 can regulate the proliferation,migration,and tube formation ability of HRMEC under high-glucose condition,and also inhibit the overexpression of VEGFA and MMP9,which may be through regulating the classical Wnt/β-catenin pathway.

Objective To explore the characteristics of apolipoprotein E(APOE)gene polymorphism in patients with Alzheimer's disease(AD)and mild cognitive impairment(MCI)due to AD,as well as its corre-lation with baseline levels of ketone bodies.Methods A total of 110 AD patients from the outpatient and neu-rology wards of the hospital from January 2020 to October 2023 were selected as the AD group,105 patients(none of whom had used anti dementia drugs)were selected as the MCI group,and 110 healthy elderly exami-nees in the physical examination center were selected as the control group.APOE gene polymorphism,and the levels of serum β-hydroxybutyrate(HB)and urine ketone bodies were measured.The distribution of APOE genotype among the three groups was analyzed,and the differences of the levels of serum HB and urine ketone bodies were compared among those carried APOE ε4 allele and those did not.Results Among the three groups,the statistical significance was found in the differences of APOE genotype and ε2,ε3,ε4 allele(P<0.05).The proportion of APOE ε4 allele carriers in the AD group and the MCI group was higher than that in the control group(P<0.05).The levels of serum βHB in the AD group and the MCI group were lower than that in the control group(P<0.05).The levels of serum βHB in those carried APOE ε4 in the AD group were significantly lower than that in the control individuals(P<0.05).There was no statistically significant differ-ence in serum βHB levels between individuals carried and not carried APOE ε4 in the three groups(P>0.05).There was no statistically significant difference in the levels of urinary ketones among the three groups(P>0.05).There was no statistically significant difference in urine ketone bodies levels between individuals carried and not carried APOE ε4 in the three groups(P>0.05).Conclusion The reduced baseline levels of serum βHB in AD patients are associated with APOE ε4 allele.

Objective To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly(VM)in fetuses.Methods Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA,and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups.Three miRNAs with significant differential expression between the two groups,whose high expression was associated with VM,were selected for verification with RT-qPCR.Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes AKT3 and CCDC88C.Gene ontology(GO)and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.Results We identified a total of 272 differentially expressed miRNAs in VM cases,including 43 up-regulated and 229 down-regulated miRNAs.The target genes of these differential miRNAs were associated with DNA and transcription factor binding,transmembrane transporter and nucleic acid binding transcription factor activity,and cell developmental process.These miRNAs were mostly enriched in the MAPK,cGMP-PKG and Wnt signaling pathways.Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group(P<0.05),which was consistent with miRNA sequencing results;let-7b-5p expression level was significantly lower in VM group,which was contrary to miRNA sequencing result.Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating AKT3 or CCDC88C expressions.Conclusions The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK,PI3K-Akt,Wnt and cGMP-PKG signaling pathways.

Background: and Purpose X-linked Charcot-Marie-Tooth disease type 1 (CMTX1) is characterized by peripheral neuropathy with or without episodic neurological dysfunction. We performed clinical, neuropathological, and genetic investigations of a series of patients with mutations of the gap-junction beta-1 gene (GJB1) to extend the phenotypic and genetic description of CMTX1. Methods: Detailed clinical evaluations, sural nerve biopsy, and genetic analysis were applied to patients with CMTX1. Results: We collected 27 patients with CMTX1 with GJB1 mutations from 14 unrelated families. The age at onset (AAO) was 20.9±12.2 years (mean±standard deviation; range, 2–45 years). Walking difficulties, weakness in the legs, and pes cavus were common initial symptoms. Compared with female patients, males tended to have a younger AAO (males vs. females=15.4±9.6 vs. 32.0±8.8 years, p=0.002), a longer disease course (16.8±16.1 vs. 5.5±3.8 years, p=0.034), and more-severe electrophysiological results. Besides peripheral neuropathy, six of the patients had special episodic central nervous system (CNS) evidence from symptoms, signs, and/or reversible white-matter lesions. Neuropathology revealed the loss of large myelinated fibers, increased number of regenerated axon clusters with abnormally thin myelin sheaths, and excessively folded myelin. Genetic analysis identified 14 GJB1 variants, 6 of which were novel. Conclusions These findings expand the phenotypic and genetic spectrum of CMTX1. Although CMTX1 was found to have high phenotypic and CNS involvement variabilities, detailed neurological examinations and nerve conduction studies will provide critical clues for accurate diagnoses. Further exploration of the underlying mechanisms of connexin 32 involvement in neuropathy or CNS dysfunction is warranted to develop promising therapies.

Humans ; Phosphorylation ; Serine/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Signal Transduction ; Neovascularization, Pathologic

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40％of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated he-patic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and ther-apeutic targets for liver protection.

Objective:To provide a new morning check method for the output dose stability of the multileaf collimator (MLC) of the CyberKnife M6 (CK-M6) system.Methods:The CT images of a verification phantom with a size of 20 cm × 20 cm × 10 cm were transmitted into the Precision Treatment Plan ning System (ver. 1.1.1.1). The high-precision alignment between the accelerator output front and the fixed position of the phantom surface was achieved using the fiducial tracking method. A 10 cm × 10 cm radiation field was formed by the MLC and a DailyCheck plan with an output of 200 MU was designed. The repeatability, sensitivity, and accuracy of the DailyCheck plan were measured, and the CK-M6 system was continuously tested for one month using the artificial fixed method and the DailyCheck plan designed in this study. Results:The average and the standard deviation of 10 repeated measurements by the DailyCheck plan were 492.28 pC and 0.09, respectively, indicating good stability. There was a linear correlation between the measured values and the output dose, with a correlation coefficient of R2 > 0.999. Moreover, there was a position deviation of 2 mm between the phantom and the accelerator output front, and the result ant effect on the measured values was equivalent to a dose deviation caused by an output of 1.24 MU. The result from the continuous measurement of both the artificial fixed method and the DailyCheck plan fell within permissible limits, showing high consistency. Conclusions:The DailyCheck plan established through the fiducial tracking of a verification phantom can achieve the convenient, quick, and accurate daily detection of the output dose stability of the MLC of CK-M6. Therefore, this method can be widely applied in the clinical quality control of the CK-M6 system.

Objective: To explore the interrelationships among fathers’ parenting effectiveness, urban children’s sociality and the acceptance of “second-born children” by children (including only children and “first-born children”), and to provide a reference for interventional measured for improving children’s acceptance rate of "second-bron children". Methods: Stratified cluster sampling method was used to select 3 to 9-year-old 778 only children and “first-born children” in two kindergartens and grades 1-3 of two primary schools in a city, and the children and their fathers were investigated with questionnaires survey regarding general information of both fathers and children, the acceptance degree of “second-born children”, the sociality of children and the parenting effectiveness of fathers. Results: Children in different grades had different acceptance levels of “second-born children” and the difference was statistically significant (F=2.89，P<0.05). Correlation analysis showed that the acceptance of “second-born children” was negatively correlated with the interaction between the only children and “first-born children”, and positively correlated with the prosocial behavior of the only children and “first-born children”, both of which are statistically significant (r=-0.10，0.10,P<0.01). Father’s role efficacy was negatively correlated with onlychildren and “first-born-children” peer interaction problems(r=-0.15), and positively correlated with prosocial behavior(r=0.32), both of which were statistically significant (P<0.01). The structural equation model of father’s parenting effectiveness, children’s sociality and acceptance of “second-born children” showed that low father’s parenting effectiveness could positively predict children’s peer communication problems and negatively predict children’s prosocial behavior, and thus children’s peer communication problems could negatively predict the acceptance degree of only children and “first-born children” to “second-born children”, and prosocial behaviors could positively predict the acceptance degree of “second-born children”. Conclusion Promoting father’s parenting efficiency can facilitate the social development of children and the acceptance of siblings.It’s recommended that fathers actively participate in specific parenting matters for the only child and "one child",and consciously improve parenting effectiveness.