OBJECTIVE: To delineate the clinical and genetic features of a Chinese girl harboring a rare de novo variant of SYNGAP1 associated with Mental retardation, autosomal dominant 5 (MRD5), and to conduct a comprehensive genotype-phenotype correlation analysis within the Chinese population through an extensive literature review. METHODS: A 5-year-old girl presenting with seizures without an obvious cause was enrolled in September 2020. Genomic DNA was extracted from the patient and her parents. Whole exome sequencing (WES) was performed on the proband to identify suspected pathogenic variants based on her clinical phenotype. Sanger sequencing was used for validation, followed by bioinformatic analysis of the variant. Additionally, data from 54 previously reported Chinese cases with SYNGAP1 variants were integrated to summarize the distribution of variant types and clinical characteristics. Ethical approval was obtained from the Ethics Committee of Kunming Children's Hospital (Ethics No.: 2021-03-055-K01). RESULTS: WES identified a heterozygous nonsense variant, SYNGAP1 c.725G>A (p.Trp242*), in the proband. Sanger sequencing confirmed it was a de novo variant. According to the ACMG guidelines, this variant was classified as pathogenic (PVS1+PS2). Based on the clinical manifestations, the patient was diagnosed with MRD5. Bioinformatic analysis suggested that this variant introduces a premature stop codon at tryptophan 242, disrupting the PH domain and leading to the loss of the C2, Ras-GAP, and C-terminal domains. The pooled analysis of Chinese cases revealed that nonsense (38.2%) and frameshift (36.4%) variants were the predominant types. Intellectual disability/developmental delay was present in 100.0% of patients, epilepsy in 83.6%, and autism spectrum disorder in 41.3%. The incidence of epilepsy differed significantly among variant types (P = 0.045). Exons 8 and 15 were identified as mutation hotspots. CONCLUSION This study has identified a SYNGAP1 c.725G>A variant in the Chinese population and confirmed it as a potential cause of MRD5, which expanded the mutational spectrum of this disorder.
Humans ; Female ; Child, Preschool ; Intellectual Disability/genetics* ; ras GTPase-Activating Proteins/genetics* ; Phenotype ; Exome Sequencing ; Genetic Association Studies

Obesity and its related metabolic diseases have become a major global public health threat, and its rising incidence significantly increases the risk of cardiovascular and cerebrovascular diseases, diabetes and other complications. Pancreatic lipase is a key enzyme that converts food-borne lipids into triglycerides and fatty acids, and the effective inhibition of its activity has become an important strategy for the treatment of obesity. This paper discusses the screening methods of pancreatic lipase inhibitors, and summarizes and reviews the basic principles, advantages and disadvantages and application status of traditional screening methods, modern new screening methods and virtual screening methods. In view of the problems faced by the screening methods of pancreatic lipase inhibitors, future research urgently needs to move towards a collaborative innovation path of multi-technology integration, intelligent screening and complex systematization of traditional Chinese medicine, so as to open up new research paradigms.

Objective To explore a method for rapid and differential diagnosis of rejection after kidney transplantation through urine hyperspectral imaging technology. Methods Hyperspectral data information from urine samples of 118 recipients after kidney transplantation was collected, and a deep learning model was constructed to diagnose and classify the types of rejection. Results A deep learning diagnostic model based on the 34-layer residual network (ResNet-34) was constructed, and 118 patients were included and divided into the training set and the test set. Based on the pathological results of the transplanted kidney puncture, the urine samples of the patients were classified into five groups: the non-rejection group, the T-cell-mediated rejection group, the antibody-mediated rejection group, the mixed rejection group and the nephropathy recurrence group. The results showed that the diagnostic sensitivities of the model for the above five groups were 0.960, 0.980, 0.930, 0.940 and 0.943 respectively, and the diagnostic specificities were 0.983, 0.993, 0.997, 0.989 and 0.989 respectively. The overall diagnostic accuracy rate reached 95.7%. Conclusions The study provides a non-invasive, rapid and accurate auxiliary diagnostic method for the differential diagnosis of rejection after kidney transplantation.

AIM:To investigate the effect of paeoniflorin on the inflammatory response of diabetic retinopathy rats by regulating phosphatidylinositol-3 kinase/protein kinase B(PI3K/Akt)signaling pathway.METHODS: A total of 70 SPF male SD rats were selected, and 12 rats were randomly selected as the control group(normal saline gavage). The remaining 58 rats were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ)to establish diabetic rat models. Rats with diabetic retinopathy were randomly divided into model group(normal saline), paeoniflorin low-dose group(100 mg/kg paeoniflorin), paeoniflorin high-dose group(200 mg/kg paeoniflorin)and metformin group(100 mg/kg metformin), with 12 rats in each group. The body mass of the rats in each group were compared. HE staining was used to observe the pathological changes of the rat retina. Automatic biochemical analyzer was used to detect the levels of fasting blood glucose, glycosylated hemoglobin, serum high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), total cholesterol and triglyceride in the rats. Enzyme-linked immunosorbent assay was used to detect the levels of serum superoxide dismutase(SOD), reactive oxygen species(ROS), malondialdehyde(MDA), glutathione peroxidase(GSH-PX), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)and interleukin-1β(IL-1β)in the rats. Western blot was used to detect the expressions of Occludin, p-PI3K, tight junction protein-1(ZO-1), p-Akt and VE-Cadherin in the rat retina.RESULTS: The expression levels of Occludin, ZO-1 and VE-cadherin in low-dose and high-dose paeoniflora groups were higher than those in the model group, while the expression levels of TNF-α, IL-6, IL-1β, p-PI3K and p-Akt in serum were lower than those in the model group. The high-dose group of paeoniflorin was significantly better than the low-dose group of paeoniflorin(all P<0.05).CONCLUSION: Paeoniflorin may reduce inflammatory response in diabetic retinopathy rats by inhibiting PI3K/Akt signaling pathway.

AIM:To investigate the effect of paeoniflorin on the inflammatory response of diabetic retinopathy rats by regulating phosphatidylinositol-3 kinase/protein kinase B(PI3K/Akt)signaling pathway.METHODS: A total of 70 SPF male SD rats were selected, and 12 rats were randomly selected as the control group(normal saline gavage). The remaining 58 rats were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ)to establish diabetic rat models. Rats with diabetic retinopathy were randomly divided into model group(normal saline), paeoniflorin low-dose group(100 mg/kg paeoniflorin), paeoniflorin high-dose group(200 mg/kg paeoniflorin)and metformin group(100 mg/kg metformin), with 12 rats in each group. The body mass of the rats in each group were compared. HE staining was used to observe the pathological changes of the rat retina. Automatic biochemical analyzer was used to detect the levels of fasting blood glucose, glycosylated hemoglobin, serum high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), total cholesterol and triglyceride in the rats. Enzyme-linked immunosorbent assay was used to detect the levels of serum superoxide dismutase(SOD), reactive oxygen species(ROS), malondialdehyde(MDA), glutathione peroxidase(GSH-PX), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)and interleukin-1β(IL-1β)in the rats. Western blot was used to detect the expressions of Occludin, p-PI3K, tight junction protein-1(ZO-1), p-Akt and VE-Cadherin in the rat retina.RESULTS: The expression levels of Occludin, ZO-1 and VE-cadherin in low-dose and high-dose paeoniflora groups were higher than those in the model group, while the expression levels of TNF-α, IL-6, IL-1β, p-PI3K and p-Akt in serum were lower than those in the model group. The high-dose group of paeoniflorin was significantly better than the low-dose group of paeoniflorin(all P<0.05).CONCLUSION: Paeoniflorin may reduce inflammatory response in diabetic retinopathy rats by inhibiting PI3K/Akt signaling pathway.

Aim To investigate the effect of hypobaric hypoxia on macrophage necroptosis and atherosclerotic plaque instability and explore the underlying mechanisms.Methods Mouse bone marrow-derived macrophages were i-solated and cultured,and divided into control group(21％oxygen concentration)and hypoxia group(3％oxygen concen-tration).After 48 hours,cell necroptosis was detected,and the expression of cell necroptosis related proteins was deter-mined by Western blot.Healthy male ApoE-/-mice were randomly divided into control group and hypobaric hypoxia group.After the intervention for 16 weeks,the plasma lipids and inflammatory cytokines were measured,the areas of ath-erosclerotic plaque and necrotic core were evaluated by HE staining.The content of plaque collagen was detected by Mas-son staining.The number of macrophages in the plaque and the expression of necrotic apoptosis related proteins were de-tected by immunohistochemical staining and Western blot.Results Hypoxia induced increased necrotic apoptosis of macrophages(P＜0.01),while necroptotic inhibitor necrostatin-1(Nec-1)reduced hypoxia induced cell death(P＜0.05);hypoxia leads to a decrease in the expression of adenosine deaminase acting on RNA 1(ADAR1)in macrophages(P＜0.01),and an increase in the expression of Z-DNA binding protein 1(ZBP1),phosphorylated receptor-interacting serine/threonine-protein kinase(p-RIPK3),and phosphorylated mixed lineage kinase domain-like protein(p-MLKL)(all P＜0.01).Compared with the control group,the plasma lipid levels of ApoE-/-mice in the hypobaric hypoxia group did not change significantly(P＞0.05),the plasma inflammatory cytokines(TNF-α,IL-1β,IL-6 and MCP-1)increased(all P＜0.05),the area of atherosclerotic plaque increased(P＜0.05),the area of plaque necrotic core increased,the content of plaque collagen decreased,the number of macrophages increased,the expression of ADAR1 decreased,and the expres-sion of ZBP1 and p-MLKL increased(all P＜0.01).Conclusion Hypobaric hypoxia causes the imbalance of A-DAR1/ZBP1 expression in macrophages,activates RIPK3/MLKL signaling pathway,promotes macrophage necroptosis,in-creases the area of plaque necrosis core,and leads to increase instability of atherosclerotic plaque.

OBJECTIVE To establish a interview outline design process and quality control evaluation method based on grounded theory,providing ideas for qualitative research interview outline design in medical fields.METHODS A literature review was conducted to understand the current research status;a preliminary interview outline was developed around the research content.The triangulation method,group evaluation,expert review and pre-interview were adopted to execute the interview outline and conduct quality control.The evaluation indicators and target values were formulated(an average score for the overall quality evaluation of all indicators ≥4.5,and an average score for individual indicators ≥ 4.00)to evaluate the effect of the interview outline.Taking the research on the mechanism of physicians'prescribing behavior under the background of Diagnosis Related Groups(DRGs)payment as an example,the methodological contents of above interview outline were applied in practical research.RESULTS The interview outline included basic information and interview questions.The interview questions were divided into three parts:influencing factors survey,promoting and hindering factors of standardizing physician prescription behavior,and communication,with a total of 12 questions.After being reviewed by members of the research group,experts review and pre-interview,a total of 9 people participated in the quality control evaluation of the interview outline.The overall evaluation score was 4.94(＞4.50),and the average score of each indicator was greater than 4.00,indicating that the quality of the outline met the requirements for the interview and could be used for the formal interview.CONCLUSIONS The established interview outline design and quality control method based on grounded theory provides ideas for the qualitative research interview outline design in the medical field,and lays the foundation for further using grounded theory to study the influencing factors and mechanisms of physician prescription behavior under the DRG background.

Objective:To explore the effect of β-galactoside α-2-6sialyltransferase1(ST6GAL1)on glycolysis,migration and invasion of colorectal cancer(CRC)HCT116 cells and its possible molecular mechanisms.Methods:The difference in the expression of ST6GAL1 in CRC patients and healthy people was analyzed using the GEPIA2 database.WB was performed to detect the differences in the expressions of ST6GAL1 in CRC cell lines HCT116,SW480,Caco-2,HT29,LoVo and human normal colon epithelial cell line NCM460.The difference in the expressions of ST6GAL1 in CRC tissues and corresponding adjacent tissues was analyzed by immunohistochemistry.HCT116 cell lines with stably knocked down or overexpressed ST6GAL1 were constructed by lentivirus transfection.Cell migration ability was detected by scratch test.Cell invasion ability was detected by Transwell test.WB assay was performed to detect the expression levels of cell glycolysis-related proteins and Notch1 intracellular domain(Notch1 ICD)as well as the phosphorylation level of PI3K/AKT/mTOR pathway.The expression level of Notch1 ICD and its entry into nucleus were observed by immunofluorescence assay.The Notch1 receptor agonist Jagged1 was added to HCT116 cells,and the expression levels of glycolysis-related proteins and Notch1 ICD and PI3K/AKT/mTOR pathway phosphorylation level were detected by WB.Results:The expression of ST6GAL1 was up-regulated in CRC tissues and cells(all P<0.05).Compared with the control and overexpression groups,knockdown of ST6GAL1 resulted in significantly lower levels of Notch1 ICD expression and PI3K/AKT/mTORC1 phosphorylation in HCT116 cells,lower levels of cellular glycolysis-related protein expressions and weaker cell migration and invasion abilities(all P<0.05).Overexpression of ST6GAL1 increased Notch1 ICD expression levels within HCT116 cells and promoted their entry into the nucleus.Cell glycolysis-related protein expression levels were elevated(all P<0.05).Cell migration and invasion abilities were enhanced(all P<0.05).Conclusion:ST6GAL1 activates the PI3K/AKT/mTORC1 pathway through activation of Notch1 receptor and phosphorylation,thus enhancing the glycolytic level and migration and invasion abilities of CRC cells.

Objective To explore the value of MRI subtraction technique(ST)for evaluating the efficacy of systemic therapy for advanced hepatocellular carcinoma(HCC)and predicting prognosis after combining with surgery.Methods Totally 35 patients with 39 HCC lesions who received systemic therapy+radical resection were retrospectively collected.Based on preoperative MRI,tumor activity ratio(recorded as tumor activityST)was obtained with ST,while tumor activity value(recorded as tumor activitypathology)was obtained through postoperative pathology,and their correlation was analyzed.The patients were regularly followed up after surgery,and the survival data were recorded.Receiver operating characteristic(ROC)curve was drawn to evaluate the efficacy of tumor activityST for predicting patients'survival status.Then the patients were divided into survival benefit group and no survival benefit group according to the cut-off value,and survival analysis was conducted.Results Tumor activityST was positively correlated with tumor activitypathology(r=0.900,P＜0.001).The median follow-up time was 32.93 months,during which 8 patients died,and the median survival time was 29.9 months.The area under the curve(AUC)of tumor activityST for predicting patients'survival status was 0.67,and the cut-off value was 0.36.Thirty patients with tumor activityST＜0.36 were enrolled in survival benefit group,while 5 patients≥0.36 were collected in no survival benefit group.The overall survival in survival benefit group was longer than that in no survival benefit group(P＜0.001).Conclusion MRI ST could be used to non-invasively evaluate the efficacy of systemic therapy for advanced HCC and predict prognosis after combining with surgery.

Objective To study whether electroacupuncture(EA)attenuates traumatic brain injury(TBI)by down-regulating glutamate carboxypeptidase Ⅱ(GCPII)gene expression in rats.Methods Rats were divided into Sham group(n=10),TBI group(n=11),TBI+EA group(n=11),TBI+EA+GCPII-NC group(n=11)and TBI+EA+GCPII-OE group(n=11).Sham group rats underwent sham surgery,while other groups rats underwent TBI modeling using an electronic brain injury instrument.The rats in Sham group and TBI group were fed normally,and the rats in other groups were treated with electroacupuncture for 14 days.On the basis of electroacupuncture treatment,rats in TBI+EA+GCPII-NC group and TBI+EA+GCPII-OE group were injected with GCPII-NC and GCPII-OE,respectively.After treatment,the cognitive function of rats was evaluated by neurological function score and Morris water labyrinth task.HE and TUNEL staining were performed on brain tissue.The water content,glutamate(Glu)content,calcium(Ca2+)concentration,superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in brain tissue of rats in each group were measured.The mRNA or protein expression of GCPII,Bax,Bcl2 and cleaved caspase-3 in brain tissue were detected by qRT-PCR or Western blot.Results Compared with Sham group,the expression of mRNA and protein of GCPII in TBI group increased,the neurological function score and escape latency increased,the number of crossing the platform decreased,the water content increased,the cortex showed obvious damage,the content of Glu increased,the concentration of calcium increased,the levels of SOD,CAT and GSH-Px decreased,the level of MDA increased,the positive rate of TUNEL increased,the expression of Bax and cleaved caspase-3 protein increased,and the expression of Bcl2 protein decreased significantly(P<0.05).Compared with TBI group,the expression level of mRNA and protein of GCPII in TBI+EA group decreased,the neurological function score and escape latency decreased,the number of crossing platform increased,the water content decreased,the morphology of cortex improved obviously,the content of Glu decreased,the concentration of calcium decreased,the level of SOD,CAT and GSH-Px increased,the level of MDA decreased,the positive rate of TUNEL decreased,the expression level of Bax and cleaved caspase-3 protein decreased,and the expression level of Bcl2 protein increased significantly(P<0.05).Compared with TBI+EA group and TBI+EA+GCPII-NC group,the expression level of mRNA and protein of GCPII in TBI+EA+GCPII-OE group increased,neurological function score and escape latency increased,the number of crossing platform decreased,water content increased,cortical injury aggravated,Glu content increased,calcium concentration increased,SOD,CAT and GSH-Px levels decreased,MDA level increased,TUNEL positive rate increased,Bax and cleaved caspase-3 protein expression increased,and the expression of Bcl2 protein decreased significantly(P<0.05).Conclusion By down-regulating the expression of GCPII gene,electroacupuncture reduced the content of glutamate in the brain tissue of TBI rats,thereby inhibiting calcium overload,oxidative stress and neuronal apoptosis in the brain tissue,and thereby improving cognitive function and alleviating brain injury.