Objective:To explore the epileptic phenotype spectrum of SLC6A1 gene mutations and their genotype-phenotype correlation. Methods:Four hundred patients with epilepsy of unknown etiology admitted to Epilepsy Center, Department of Neurology, Second Affiliated Hospital of Guangzhou Medical University from July 2019 to July 2024 were enrolled to screen the SLC6A1 gene mutations; the clinical characteristics, mutation pathogenicity, and changes of hydrogen bond between amino acids, stability and amino acid hydrophobicity of SLC6A1 gene encoded proteins caused by missense mutations in patients with SLC6A1 gene mutations were analyzed. At the same time, a comprehensive search was conducted in PubMed, HGMD and CNKI databases to collect the publicly reported SLC6A1 gene mutations related to epilepsy up to September 8, 2024; differences in proportion of missense mutations between the two most common and featured epileptic phenotypes and proportion of missense mutations in loops of SLC6A1 gene coding proteins were analyzed. Results:Five patients carried SLC6A1 gene mutations in 400 patients with epilepsy of unknown etiology: 2 had de novo heterozygous canonical splice site mutations (c.850-1G>A and c.1324-1G>A), with phenotypes as partial epilepsy combined with severe development delay and childhood absence epilepsy combined with mild developmental delay; 2 had de novo heterozygous missense mutations (c.187G>A/p.Gly63Ser and c.1081C>A/p.Pro361Thr), with phenotypes as partial epilepsy combined with mild development delay and generalized epilepsy combined with severe development delay; and one had heterozygous missense mutation of unknown origin (c.700G>A/p.Gly234Ser), with phenotype as Lennox-Gastaut syndrome. Four de novo mutations were evaluated as having pathogenic or likely pathogenic features, and one mutation of unknown origin was evaluated as of uncertain significance. In addition, 3 missense mutations caused significant changes in number or bonding form of hydrogen bonds between amino acids of the encoded proteins, with obviously decreased stability and hydrophobicity of the encoded proteins. (2) Results of literature analysis showed that 84 SLC6A1 mutations have been reported to be associated with epilepsy; combined with the genetic results in this study, a total of 89 SLC6A1 mutations were identified, including 53 missense mutations, 33 nonsense mutations, and 3 in-frame/in-del mutations; 7 epilepsy phenotypes were involved, including 38 patients with myoclonic atonic epilepsy (MAE), 16 with epilepsy, 12 with epileptic encephalopathy, 8 with childhood absence epilepsy, 6 with childhood-onset epilepsy, 6 with generalized epilepsy, and 3 with focal epilepsy. No significant difference in proportion of missense mutations was noted between MAE and epileptic encephalopathy patients ( P>0.05); however, the proportion of missense mutations in loops of the epileptic encephalopathy patients was significantly higher than that of the MAE patients ( P<0.05). Conclusion:SLC6A1 gene mutations can cause complex and diverse epilepsy phenotype spectrum, and most patients are accompanied by developmental delay; subregional effect of the encoded protein molecules may be a potential mechanism for different clinical phenotypes between MAE and epileptic encephalopathy caused by SLC6A1 gene mutations.

Objective:To evaluate the role of computed tomography (CT) imaging features in predicting the risk classification of duodenal stromal tumor (DST), so as to assist clinical treatment decision-making.Methods:A total of 109 patients with DST confirmed by surgical pathology at Zhongshan Hospital, Fudan University were retrospectively enrolled from November 1, 2013 to November 30, 2022. According to the National Institutes of Health risk classification criteria, the 109 DST patients were divided into low-risk group (including very low- and low-risk patients, 70 cases) and high-risk group (including moderate- and high-risk patients, 39 cases). The CT features of the 2 groups were collected, including arterial and venous phase CT values, tumor size index, shape (regular or not), edge (clear or not), growth pattern (intracavity, extracavity or both), degree of cystic necrosis (0 degree, Ⅰ degree, Ⅱ degree, Ⅲ degree), and the presence of enlarged feeding arteries and vascular-like enhancement were collected. Univariate and multivariate logistic regression analyses were performed to identify the CT features related to DST risk classification. Receiver operating characteristic curve (ROC) was drawn to assess the predictive value of the features.Results:The result of univariate logistic regression analysis showed that tumor size index ( P<0.001), arterial phase CT value ( P<0.001), venous phase CT value ( P<0.001), shape ( P<0.001), edge ( P=0.004), growth pattern ( Pintracavity with extracavity=0.004), degree of cystic necrosis ( PⅡ degree=0.003, PⅢ degree=0.002), enlarged feeding arteries ( P=0.001), and vascular-like enhancement ( P=0.004) were all related to DST risk classification. The result of multivariate logistic regression analysis revealed that tumor size index was an independent risk factor for predicting DST risk classification ( OR=1.35, 95% confidence interval (95% CI) 1.10 to 1.65, P=0.003). The result of ROC analysis demonstrated that the area under the curve for tumor size index in predicting DST risk classification was 0.909 (95% CI 0.841 to 0.977), with sensitivity of 82.1% and specificity of 92.9%. Conclusion:Tumor size index based on CT imaging has good predictive performance for DST risk classification, and provides valuable assistance for clinical diagnosis and treatment decisions.

Objective:To evaluate the role of computed tomography (CT) imaging features in predicting the risk classification of duodenal stromal tumor (DST), so as to assist clinical treatment decision-making.Methods:A total of 109 patients with DST confirmed by surgical pathology at Zhongshan Hospital, Fudan University were retrospectively enrolled from November 1, 2013 to November 30, 2022. According to the National Institutes of Health risk classification criteria, the 109 DST patients were divided into low-risk group (including very low- and low-risk patients, 70 cases) and high-risk group (including moderate- and high-risk patients, 39 cases). The CT features of the 2 groups were collected, including arterial and venous phase CT values, tumor size index, shape (regular or not), edge (clear or not), growth pattern (intracavity, extracavity or both), degree of cystic necrosis (0 degree, Ⅰ degree, Ⅱ degree, Ⅲ degree), and the presence of enlarged feeding arteries and vascular-like enhancement were collected. Univariate and multivariate logistic regression analyses were performed to identify the CT features related to DST risk classification. Receiver operating characteristic curve (ROC) was drawn to assess the predictive value of the features.Results:The result of univariate logistic regression analysis showed that tumor size index ( P<0.001), arterial phase CT value ( P<0.001), venous phase CT value ( P<0.001), shape ( P<0.001), edge ( P=0.004), growth pattern ( Pintracavity with extracavity=0.004), degree of cystic necrosis ( PⅡ degree=0.003, PⅢ degree=0.002), enlarged feeding arteries ( P=0.001), and vascular-like enhancement ( P=0.004) were all related to DST risk classification. The result of multivariate logistic regression analysis revealed that tumor size index was an independent risk factor for predicting DST risk classification ( OR=1.35, 95% confidence interval (95% CI) 1.10 to 1.65, P=0.003). The result of ROC analysis demonstrated that the area under the curve for tumor size index in predicting DST risk classification was 0.909 (95% CI 0.841 to 0.977), with sensitivity of 82.1% and specificity of 92.9%. Conclusion:Tumor size index based on CT imaging has good predictive performance for DST risk classification, and provides valuable assistance for clinical diagnosis and treatment decisions.

Detailed characterizations of genomic alterations have not identified subtype-specific vulnerabilities in adult gliomas. Mapping gliomas into developmental programs may uncover new vulnerabilities that are not strictly related to genomic alterations. After identifying conserved gene modules co-expressed with EGFR or PDGFRA (EM or PM), we recently proposed an EM/PM classification scheme for adult gliomas in a histological subtype- and grade-independent manner. By using cohorts of bulk samples, paired primary and recurrent samples, multi-region samples from the same glioma, single-cell RNA-seq samples, and clinical samples, we here demonstrate the temporal and spatial stability of the EM and PM subtypes. The EM and PM subtypes, which progress in a subtype-specific mode, are robustly maintained in paired longitudinal samples. Elevated activities of cell proliferation, genomic instability and microenvironment, rather than subtype switching, mark recurrent gliomas. Within individual gliomas, the EM/PM subtype was preserved across regions and single cells. Malignant cells in the EM and PM gliomas were correlated to neural stem cell and oligodendrocyte progenitor cell compartment, respectively. Thus, while genetic makeup may change during progression and/or within different tumor areas, adult gliomas evolve within a neurodevelopmental framework of the EM and PM molecular subtypes. The dysregulated developmental pathways embedded in these molecular subtypes may contain subtype-specific vulnerabilities.
Humans ; Brain Neoplasms/pathology* ; Neoplasm Recurrence, Local/metabolism* ; Glioma/pathology* ; Neural Stem Cells/pathology* ; Oligodendrocyte Precursor Cells/pathology* ; Tumor Microenvironment

Objective:To investigate the clinical characteristics and outcome of novel coronavirus pneumonia (COVID-19)patients with different body mass index (BMI), and to provide the basis for disease assessment and prognosis.Methods:The clinical data of 541 patients with COVID-19 diagnosed in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from January 16 to March 28, 2020 were collected. The patients were divided into normal weight group, overweight group, and obesity group according to BMI. The clinical characteristics and outcomes of the three groups were compared. The correlation between BMI and clinical classification was analyzed by ordinal logistic regression.Results:There were 288 cases (53.23%) in normal weight group, 193 cases (35.67%) in overweight group, and 60 cases (11.09%) in obesity group. Compared with normal weight group, overweight and obesity groups displayed higher proportion of hypertension, with increased levels of white blood cells, neutrophils, C reactive protein, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and triglyceride in laboratory test results, and higher proportion of severe and critical illness ( P<0.05 or P<0.01). After adjusting for age, gender, and underlying diseases, regression analysis showed that higher BMI predicted more severe clinical classification ( OR=1.079, 95% CI 1.010-1.154). Conclusion:In COVID-19 patients, overweight and obese patients are more likely to develop into severe and critical illness, suggesting that obesity may be an important risk factor affecting the clinical outcome of COVID-19.

Objective: To investigate the influence of extremely low-frequency magnetic field on periodical expression of cryptochrome (Cry) gene in mouse embryonic fibroblast NIH3T3 cells. Methods: The NIH3T3 cells were divided into magnetic field group and sham-exposure group. The NIH3T3 cells in the magnetic field group were stimulated by horse serum and then exposed to an extremely low-frequency magnetic field (50 Hz and 0.3 mT) for 48 hours, and those in the sham-exposure group were also stimulated by horse serum and then exposed to a coil for 48 hours. The NIH3T3 cells were collected, total RNA was extracted, and cDNA was obtained via reverse transcription. Real-time fluorescent quantitative RT-PCR was used to measure the changes in transcription cycles of Cry and Period genes in both groups. Results: There was no significant difference in the proliferation rate at 0, 12, 24, and 48 hours of exposure between the two groups (P>0.05) . Both sham-exposure group and magnetic field group showed a rhythmic change in the expression of Cry gene, and compared with the sham-exposure group, the magnetic field group had a significantly shortened circadian rhythm of Cry gene in NIH3T3 cells (t=2.57, P<0.05) . Both groups had rhythmic and periodical expression of Period gene and there was no significant difference between the two groups (t=0.70, P>0.05) . Conclusion Extremely low-frequency magnetic field can significantly shorten the circadian rhythm of Cry gene in mouse embryonic fibroblasts, while there is no significant change in the circadian rhythm of Period gene.

OBJECTIVE:To provide reference for the establishment of quality standard for Fresh Gastrodia elata lyophilized powder. METHODS:TLC was conducted for the qualitative identification of G. elata in preparation,and HPLC was conducted for the content determination of gastrodin in preparation. The column was Diamonsil C18 with mobile phase of acetonitrile-0.05% phos-phoric acid(3∶97,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 220 nm,column temperature was 25 ℃ and vol-ume injection was 10 μl. RESULTS:TLC pots of Fresh G. elata lyophilized powder were clear and well-separated. The linear of gastrodin was 2.9-14.5 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.00%;recov-ery was 98.07%-102.70%(RSD=1.74%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible,and suitable for the quality control of Fresh G. elata lyophilized powder.

OBJECTIVETo detect JAK2 V617F mutation burden and its clinical implications in patients with myeloproliferative neoplasm (MPN).

METHODSJAK2 V617F mutation burden were detected by using MGB Taqman probes and its clinical significance were retrospectively studied in 415 MPN patients.

RESULTSJAK2 V617F was found in 56.9% of all patients [83.5% in polycythemia vera (PV), 55.9% in essential thrombocythemia (ET), 41.9% in primary myelofibrosis (PMF) and 64.7% in MPN-unclassifiable)]. The majority of patients carried heterozygous JAK2 V617F mutation and homozygote was found only in 12 cases (4 in PV, 4 in MPN-U, 2 in PMF, 1 in ET, and 1 in chronic neutrophilic leukemia). Most patients (68.8%) were lower mutation burden (mutation burden<50%), but PV had the highest burden, the moderate burden in PMF and the least in ET. The patient's age and WBC count were significantly correlated with higher mutation burden in PV. WBC count was significantly related to higher mutation burden in ET. WBC count, Hb level and the platelet count were significantly related to higher mutation burden in PMF.

CONCLUSIONThe mutation burden of JAK2 V617F from high to low was PV, ET and PMF. The majority of JAK2 V617F mutation was heterozygous. JAK2 V617F mutation burden was positively correlated with age, WBC, Hb and platelet counts.

Homozygote ; Humans ; Janus Kinase 2 ; Leukocyte Count ; Mutation ; Myeloproliferative Disorders ; Platelet Count ; Polycythemia Vera ; Retrospective Studies ; Thrombocythemia, Essential

PURPOSE: The degradation of the extracellular matrix has been shown to play an important role in the treatment of hepatic cirrhosis. In this study, the effect of thalidomide on the degradation of extracellular matrix was evaluated in a rat model of hepatic cirrhosis. MATERIALS AND METHODS: Cirrhosis was induced in Wistar rats by intraperitoneal injection of carbon tetrachloride (CCl4) three times weekly for 8 weeks. Then CCl4 was discontinued and thalidomide (100 mg/kg) or its vehicle was administered daily by gavage for 6 weeks. Serum hyaluronic acid, laminin, procollagen type III, and collagen type IV were examined by using a radioimmunoassay. Matrix metalloproteinase-13 (MMP-13), tissue inhibitor of metalloproteinase-1 (TIMP-1), and alpha-smooth muscle actin (alpha-SMA) protein in the liver, transforming growth factor beta1 (TGF-beta1) protein in cytoplasm by using immunohistochemistry and Western blot analysis, and MMP-13, TIMP-1, and TGF-beta1 mRNA levels in the liver were studied using reverse transcriptase polymerase chain reaction. RESULTS: Liver histopathology was significantly better in rats given thalidomide than in the untreated model group. The levels of TIMP-1 and TGF-beta1 mRNA and protein expressions were decreased significantly and MMP-13 mRNA and protein in the liver were significantly elevated in the thalidomide-treated group. CONCLUSION: Thalidomide may exert its effects on the regulation of MMP-13 and TIMP-1 via inhibition of the TGF-beta1 signaling pathway, which enhances the degradation of extracellular matrix and accelerates the regression of hepatic cirrhosis in rats.
Actins ; Animals ; Carbon Tetrachloride/toxicity ; Collagen Type III/metabolism ; Down-Regulation ; Extracellular Matrix/metabolism ; Immunohistochemistry ; Immunosuppressive Agents/*pharmacology ; Liver Cirrhosis, Experimental/chemically induced/*metabolism/pathology/*prevention & control ; Male ; RNA, Messenger/analysis/metabolism ; Rats ; Rats, Wistar ; Thalidomide/*pharmacology ; Tissue Inhibitor of Metalloproteinase-1/biosynthesis/*drug effects ; Transcription Factor RelA/biosynthesis/drug effects ; Transforming Growth Factor beta1/biosynthesis/*drug effects ; Transforming Growth Factors/metabolism

OBJECTIVE:To establish the quality standard for Gastrodia elata hency-fired tablet. METHODS:TLC was used to identify the gastrodin and benzyl alcohol and determine the content of moisture,ash and extract;HPLC was used to determine the content of gastrodin and benzyl alcohol. Column was Diamonsil C18 with mobile phase of acetonitrile-0.05% phosphate(3:97,V/V) at flow rate of 1.0 ml/min,detection wavelength was 220 nm,column temperature was 25 ℃ and volume injection was 10 μl. RE-SULTS:The TLC of gastrodin and benzyl alcohol showed clear spots and good separation. The moisture content<35.0%,total ash contents<2.0% and extract content>40.0%. Linear range of gastrodin was 25.2-126.0,12.7-63.5 μg/ml(r=0.999 9),respectively;RSDs of precision,reproducibility and stability tests were lower than 2.0%;recovery was 99.49%-102.40%(RSD=1.09%,n=6), 98.75-102.63%(RSD=1.53,n=6),respectively. CONCLUSIONS:The method is simple and good reproducibility,and can be used for the quality control of Gastrodia elata hency-fired tablet.