ObjectiveTo investigate the protective effect of liver-targeted fibroblast growth factor 21 （FGF21） overexpression against CCl₄-induced acute liver injury （ALI） and its association with silent information regulator 1/glutathione peroxidase 4 （SIRT1/GPX4） expression and ferroptosis-related markers. MethodsFemale ICR mice （8 weeks old） were randomly divided into Control， CCl₄， adeno-associated virus （AAV）-FGF21， and AAV-FGF21 + CCl₄ groups. Liver-targeted FGF21 overexpression was achieved via tail vein injection of AAV8-FGF21 virus with green fluorescent protein （GFP） labeling. ALI was induced by intraperitoneal injection of CCl₄ （0.3 mL/kg）， with sample collection at 12 hours. Body weight and liver weight were recorded to calculate the liver weight/body weight. Alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using a biochemical analyzer. FGF21 expression in liver tissues was detected by immunofluorescence. Hepatic histopathology was evaluated by HE staining. RT-qPCR was performed to assess mRNA expression of ferroptosis and iron metabolism-related genes （ACSL4， PTGS2， HJV， FPN， GPX4）， FGF21， and SIRT1. Protein expression of FGF21， SIRT1， and GPX4 were determined by Western blot. Iron content， malondialdehyde （MDA）， glutathione （GSH）， and superoxide dismutase （SOD） activity in liver homogenate were quantified using assay kits. ResultsCompared with the Control group， the FGF21 mRNA and protein expression elevated in CCl₄ group； serum ALT/AST levels and liver weight/body weigh increased. The HE staining results showed that the liver cell membranes were ruptured. The mRNA levels of ACSL4 and PTGS2， the iron content and MDA levels all increased （P<0.01）. The mRNA levels of SIRT1， GPX4， HJV， and FPN， the GSH level and SOD activity all decreased （P<0.05）， and the protein levels of SIRT1 and GPX4 both decreased （P<0.05）. Compared with the CCl₄ group， the serum levels of ALT and AST in the AAV-FGF21 + CCl4 group were decreased （P<0.01）， and the liver/body ratio of mice decreased （P<0.01）. The results of HE staining showed that the degree of liver injury was alleviated. The mRNA levels of ACSL4 and PTGS2， iron content and MDA level all decreased （P<0.01）， while the mRNA levels of SIRT1， GPX4， HJV， FPN， GSH level and SOD activity all increased （P<0.05）. The protein levels of SIRT1 and GPX4 both increased （P<0.05）. ConclusionFGF21 overexpression is associated with enhanced SIRT1/GPX4 expression and suppression of ferroptosis features， thereby contributing to the alleviation of CCl₄-induced ALI.

Objective:To explore the early neurodevelopmental features of young children with SYNGAP1 variants and their genotype-phenotype correlation. Methods:Young children with neurodevelopmental disorders (NDDs) (< 5 years old) who were referred to the Children′s Hospital Affiliated to the Capital Institute of Pediatrics between January 2019 and July 2022 were selected as the study subjects. All children had undergone whole-exome sequencing, comprehensive pediatric neuropsychological assessment, familial segregation analysis, and pathogenicity classification. Meanwhile, young Chinese NDD children (< 5 years old) with pathogenic/likely pathogenic SYNGAP1 variants were retrieved from the literature, with information including detailed clinical and genetic testing, neurodevelopmental quotient (DQ) of the Children Neuropsychological and Behavior Scale-Revision 2016 (CNBS-R2016). Children who did not have a detailed DQ but had their developmental status assessed by a medical professional were also included. The correlation between neurodevelopmental severity, comorbidity and SYNGAP1 variants were summarized. Results:Four young NDD children carrying SYNGAP1 variants were recruited (1 male and 3 females, with a mean age of 34.0 ± 18.2 months), among whom one harboring a novel variant (c.437C>G, p. S146*). Combined with 19 similar cases retrieved from the literature, 23 Chinese NDD young children were included in our study (8 males and 10 females, 5 with unknown sex, with a mean age of 37.1 ± 14.2 months). A loss of function (LOF) variant was found in 19 (82.6%) children. All of the children had presented global developmental delay (GDD) before the age of two. In addition, 16 (69.6%) had seizure/epilepsy at the age of 27.0 ± 12.1 months, among whom 15 had occurred independent of the global developmental delay. Myoclonic and absence were common types of seizures. Compared with those with variants of exons 8 to 15, the severity of developmental delay was milder among children with variants in exons 1 to 5. Conclusion:The early neurodevelopment features of the SYNGAP1 variants for young children (< 5 years old) have included global developmental delay and seizure/epilepsy. All of the children may present GDD before the age of two. The severity of developmental delay may be related to the type and location of the SYNGAP1 variants.

Objective:To evaluate the relationship between preoperative serum bilirubin concentrations and postoperative delirium (POD) in the patients undergoing knee or hip replacement.Methods:Medical records from 413 patients undergoing knee or hip arthroplasty were selected from August 2020 to October 2023 at Qingdao Municipal Hospital using a nested case-control design based on the PNDABLE study cohort. The patients were divided into POD group ( n=77) and non-POD group ( n=336) according to whether POD occurred. Univariate analysis was used to analyze the influencing factors. Logistic regression was used to identify the risk factors for POD. The significance of mediation effect was tested. The receiver operating characteristic curve was drawn to evaluate the accuracy of risk factors in predicting POD. Results:There were significant differences in age, education time, ratio of diabetes history, Memorial Delirium Assessment Scale score, total bilirubin concentration, direct bilirubin concentration, indirect bilirubin concentration, Aβ 42 concentration, p-tau concentration, t-tau concentration, Aβ 42/p-tau ratio and Aβ 42/t-tau ratio between POD group and non-POD group ( P<0.05). The results of logistic regression analysis showed that preoperative serum total bilirubin, direct bilirubin and indirect bilirubin were risk factors for POD ( P<0.05). The results of mediation effects showed that the concentration of total tau protein in CSF partly mediated the relationship between high serum total bilirubin, direct bilirubin and indirect bilirubin concentrations and POD. The area under the receiver operating characteristic curve of total bilirubin, direct bilirubin and indirect bilirubin combined with CSF biomarker concentrations in predicting POD was 0.83 ( P<0.001). Conclusions:Preoperative elevated concentrations of total bilirubin, direct bilirubin and indirect bilirubin are risk factors for POD in the patients undergoing knee or hip replacement. CSF t-tau concentration has a partly mediating role in the association between serum total bilirubin, direct bilirubin and indirect bilirubin concentrations and the development of POD.

Abstract Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.

Objective:To explore the mechanism of epidural scar tissue hyperplasia induced by erythrocyte rupture and release of interleukin-33 (IL-33) after laminectomy in mice.Methods:In the zoological experiment, the operation group (Laminectomy) and the sham operation group were set, and HE staining and Masson staining were performed to test for blood accumulation in the operation area after laminectomy in mice. Then 12 wild-type mice with 6-8 week old were selected and divided into 4 groups: the sham operation group, the operation group (normal saline control), the pure red blood cell intervention operation group, the whole blood intervention operation group. The normal saline (100 mg/kg) was injected into the postoperative area. The red blood cells or whole blood with the same volume were injected into the postoperative area in the pure red blood cell intervention group and the whole blood intervention group. The postoperative recovery of mice in each group was observed. The levels of fibronectin in the postoperative scar tissues of mice in four groups were detected by western blot technology, and the degree of postoperative epidural scar hyperplasia was directly observed by immunohistochemistry. In the cytological experiment, the wild-type mouse erythrocyte normal saline group, the control group of IL-33 knockout mouse erythrocyte normal saline, the wild-type mouse erythrocyte lysis group, and the IL-33 knockout mouse erythrocyte lysis group were set. The levels of IL-33 in the red blood cells of four groups were detected by western blot. Then, a blank wild-type mouse erythrocyte control group, a wild-type mouse relative to the control group (only secondary antibody added to test for non-specific binding), a wild-type mouse erythrocyte group and an IL-33 knockout mouse erythrocyte group (to test for antigen specificity of the primary antibody) were set. Immunofluorescence staining was performed on the erythrocytes of four groups and the level of IL-33 was detected by flow cytometry.Results:HE staining and Masson staining after laminectomy showed that there was blood stasis in the local incision area of mice in the operation group. The epidural scar hyperplasia in the incision area of mice after whole blood or red blood cells intervention was higher, especially in the whole blood intervention group. IL-33 expression was almost undetectable in the wild-type erythrocyte normal saline control group, the IL-33-knockout erythrocyte normal saline control group, and the IL-33-knockout erythrocyte lysis group, while significant IL-33 expression was detectable in the wild-type erythrocyte lysis group. Immunofluorescence staining showed that IL-33 was expressed in and on the erythrocyte membrane of wild-type mice, while non-specific expression of IL-33 or a very small amount of IL-33 was almost undetectable in the other three groups. The immunofluorescence intensities of IL-33 in the four groups were 0.62±0.41, 60.17±4.39, 16.78±7.43 and 0.61±0.03, respectively ( F=281.90, P<0.001). The expression of IL-33 in the erythrocyte group of wild-type mice was the highest ( P<0.05). According to the results of flow cytometry, except for the trace amount of IL-33 detected in the wild-type mouse erythrocyte group, the expression of IL-33 in the other three groups was basically 0. The ratios of fibronectin to β-actin in the modeling area of the four groups gradually increased, and the ratios were 0.79±0.09, 1.26±0.23, 1.79±0.05 and 2.29±0.58, respectively, and the differences were statistically significant ( F=12.86, P=0.002). Fibronectin in the operation area of the three operation groups (normal saline control group, red blood cell intervention group and whole blood intervention group) was significantly higher than that of the sham operation group. The immunohistochemical staining results of fibronectin in the modeling area of the four groups were the same as those in western blot experiment. The average optical density values of fibronectin in each group were 0.09±0.01, 0.18±0.01, 0.22±0.01 and 0.24±0.01, respectively, and the difference was statistically significant ( F= 210.7, P<0.001). Conclusion:There is indeed blood accumulation in the surgical area after laminectomy in mice, and it can aggravate the hyperplasia of epidural scar tissue. Erythrocyte is the main component in blood, and there is a large amount of IL-33 expression in the inner and outer membrane of erythrocyte membrane. The mechanism of promoting the proliferation of epidural scar tissue may be related to the release of IL-33 by erythrocyte lysis.

Objective:To investigate the risk factors of cerebral venous sinus thrombosis (CVST) during puerperal period.Methods:This study was a restrospective analysis, A total of 33 puerperal CVST patients admitted to Beijing Tiantan Hospital, Capital Medical University from January 2016 to November 2022 were selected as the observation group, and 61 puerperal healthy women who underwent postpartum follow-up at the same period in the hospital were selected as the control group. The age, body mass index (BMI), smoking history, hypertension history, diabetes history, drug use history and mode of delivery or abortion of the two groups of women were compared and collected, as well as the level of laboratory indicators. The risk factors of CVST in puerperal period were analyzed. The measurement data of normal distribution were represented by two independent samples t test for comparison between groups. The measurement data of non-normal distribution were expressed as M ( Q1, Q3), and the Wilcoxon rank sum test was used for comparison between groups. The count data is expressed as number (%), and the comparison between groups is performed using χ 2 test or Fisher exact probability method. Logistic regression model was established to analyze the risk factors of CVST in puerperal period. Results:BMI, serum LDH, α-HBDH, fasting blood glucose, HCY levels, WBC, NEU, NLR, RDW and MPV in observation group were higher than those in control group [(30.21±4.25) kg/m 2 vs (21.94±3.02) kg/m 2]. 195.15(183.10,240.98) U/L vs 165.75(154.55,184.62) U/L, 166.60(143.10,188.60) U/L vs 124.10(116.30,137.90) U/L, (4.88 ± 0.98) vs (4.25±0.41), 8.35 (7.10, 12.16) μmol/L vs 6.60 (5.30, 7.58) μmol/L, 9.26 (6.56, 11.76) × 10 9/L vs 7.25 (6.23, 8.00) × 10 9/L, and 7.18 (4.66, 8.79) × 10 9/L vs 3.93 (3.25, 4.52) × 10 9/L, 4.13 (2.27,6.55) vs 1.63 (1.16,1.97), 42.80(38.95,47.45) fL vs 40.70(38.95,42.60) fL, (9.52±0.99) fL vs 8.96±0.88 fL], LY and PDW were lower than control group [1.58(1.11,1,96)×10 9/L vs 2.50(2.04,2.91)×10 9/L, 15.60(11.65,16.20) fL vs 16.00(15.80,16.30) fL]. The differences were statistically significant ( t=4.58, P<0.001; Z=4.54, P<0.001; Z=5.56, P<0.001; t=3.38, P=0.002; Z=4.18, P<0.001; Z=3.39, P=0.001; Z=4.92, P<0.001; Z=4.92, P<0.001; Z=4.54, P<0.001; Z=5.56, P<0.001; Z=4.18, P<0.001; Z=4.92, P<0.001; Z=5.87, P<0.001; Z=2.18, P=0.029; t=2.82, P=0.006; Z=4.78, P<0.001; Z=2.52, P=0.012). Multivariate Logistic regression analysis showed that NEU, HCY and α-HBDH were risk factors for puerparal CVST (odds ratios were 3.07, 1.53 and 1.07, respectively, 95% confidence interval: 1.65~5.71, 1.09~2.15, 1.02~1.12, P values were <0.001, 0.014, 0.007, respectively). Conclusions:α-HBDH, HCY and NEU are independent risk factors for puerperal CVST.

BACKGROUND: Recent evidence suggests that IL-33, a novel member of the IL-1b family, is involved in organ fibrosis. However, the roles of IL-33 and its receptor ST2 in epidural fibrosis post spine operation remain elusive. METHODS: A mouse model of epidural fibrosis was established after laminectomy. IL-33 in the wound tissues post laminectomy was measured with Western blotting, ELISA and imaging. The fibroblast cell line NIH-3T3 and primary fibroblasts were treated with IL-33 and the mechanisms of maturation of fibroblasts into myofibroblasts were analyzed. To explore roles of IL-33 and its receptor ST2 In vivo, IL-33 knockout (KO) and ST2 KO mice were employed to construct the model of laminectomy. The epidural fibrosis was evaluated using H&E and Masson staining, western-blotting, ELISA and immunohistochemistry. RESULTS: As demonstrated in western blotting and ELISA, IL-33 was increased in epidural wound tissues post laminectomy. The immunoflurosence imaging revealed that endothelial cells (CD31 + ) and fibroblasts (a-SAM +) were major producers of IL-33 in the epidural wound tissues. In vitro, IL-33 promoted fibroblast maturation, which was blocked by ST2 neutralization antibody, suggesting that IL-33-promoted-fibroblasts maturation was ST2 dependent. Further, IL-33/ ST2 activated MAPK p38 and TGF-β pathways. Either p38 inhibitor or TGF-β inhibitor decreased fibronectin and a-SAM production from IL-33-treated fibroblasts, suggesting that p38 and TGF-β were involved with IL-33/ST2 signal pathways in the fibroblasts maturation. In vivo, IL-33 KO or ST2 KO decreased fibronectin, a-SMA and collagen deposition in the wound tissues of mice that underwent spine surgery. In addition, TGF-β 1 was decreased in IL-33 KO or ST2 KO epidural wound tissues. CONCLUSION In summary, IL-33/ST2 promoted fibroblast differentiation into myofibroblasts via MAPK p38 and TGF-β in a mouse model of epidural fibrosis after laminectomy.

Copy number variants (CNVs) are common causes of human genetic diseases. CNVs detection has become a routine component of genetic testing, especially for pediatric neurodevelopmental disorders, multiple congenital abnormalities, prenatal evaluation of fetuses with structural anomalies detected by ultrasound. Although the technologies for CNVs detection are continuously improving, the interpretation is still challenging, with significant discordance across different laboratories. In 2020, the American College of Medical Genetics and Genomics (ACMG) and the Clinical Genome Resource (ClinGen) developed a guideline for the interpreting and reporting of constitutional copy number variants, which introduced a quantitative, evidence-based scoring framework. Here, we detailed the key points of interpreting the copy number gain based on the guideline, used six examples of different categories to illuminate the scoring process and principles. We encourage a professional understanding and application of this guideline for the detected copy number gains in China in order to further improve the clinical evaluation accuracy and consistency across different laboratories.
Child ; DNA Copy Number Variations ; Female ; Genetic Testing ; Genetics, Medical ; Genome, Human/genetics* ; Genomics ; Humans ; Pregnancy ; United States

Objective: To investigate the histopathologic, immunohistochemical, molecular genetic characteristics of dedifferentiated liposarcomas with meningothelial-like whorls(DDLMW). Methods: Six cases of DDLMW diagnosed at Jiangsu Province Hospital(the First Affiliated Hospital of Nanjing Medical University) from March 2012 to August 2018 were enrolled. The cases were analyzed by routine HE staining, immunohistochemistry(MDM2, CDK4 and p16) and fluorescent in-situ hybridization(FISH) on MDM2 gene. Related literatures were also reviewed. Results: Three of the 6 patients were male.The patient ages ranged from 40 to 77 years (mean, 58 years). Four tumors occurred in the retroperitoneum and two in the mediastinum. Histologically, the tumors showed, in addition to foci of well-differentied liposarcoma, characteristic, scattered meningothelial-like concentrical whorls. The whorls were composed of tightly, concentrically arranged, spindle to ovoid cells with mild to mederate cytological atypia. Metaplastic bone was present within or in their immediate vicinity in four case. The tumors cell also showed strong and diffuse immunoreactivity to MDM2, CDK4 and p16, but no immunoreactivity to S-100 protein, SMA, SOX10, EMA, CD21, CD23 or CD35. The Ki-67 labeling indexes were low, while FISH showed high levels of MDM2 amplification in all cases. Conclusions DDLMW is a rare morphologic variant of dedifferentiated liposarcoma. The whorls in DDLMW do not represent perineurial or follicular dendritic differentiation. Recognition and familiarity with its existence, as well as combined application of immunohistochemical staining and MDM FISH, are important to avoid confusion with other lesions.

Although high-efficiency targeted delivery is investigated for years, the efficiency of tumor targeting seems still a hard core to smash. To overcome this problem, we design a three-step delivery strategy based on streptavidin-biotin interaction with the help of c(RGDfK), magnetic fields and lasers. The ultrasmall superparamagnetic iron oxide nanoparticles (USIONPs) modified with c(RGDfK) and biotin are delivered at step 1, followed by streptavidin and the doxorubicin (Dox) loaded nanosystems conjugated with biotin at steps 2 and 3, respectively. The delivery systems were proved to be efficient on A549 cells. The co-localization of signal for each step revealed the targeting mechanism. The external magnetic field could further amplify the endocytosis of USPIONs based on c(RGDfK), and magnify the uptake distinctions among different test groups. Based on photoacoustic imaging, laser-heating treatment could enhance the permeability of tumor venous blood vessels and change the insufficient blood flow in cancer. Then, it was noticed that only three-step delivery with laser-heating and magnetic fields realized the highest tumor distribution of nanosystem. Finally, the magnetism/laser-auxiliary cascaded delivery exhibited the best antitumor efficacy. Generally, this study demonstrated the necessity of combining physical, biological and chemical means of targeting.