ObjectiveTo establish background data for a 90-day feeding trial of SD rats to ensure the reliability of research data. MethodsBackground data from six independent 90-day feeding trials of SD rats conducted by the National Center for Safety Evaluation of Drugs from 2020 to 2023 were summarized. These studies involved a blank control group of 120 SPF-grade 4-week-old SD rats, with an equal number of males and females, which were only given standard full-nutrient pelleted rat feed. After the quarantine period, the animals were observed for an additional 90 days, followed by intraperitoneal injection of Zoletil (50 mg/mL) for anesthesia, blood sampling, euthanasia, and necropsy. By analyzing the data from the blank control group, a relevant background database for SD rats was established. ResultsBoth male and female rats exhibited steady weight gain, with a more pronounced increase in male rats. Within 90 days, the average body weight of male and female rats increased to over 500 g and 300 g, respectively. Three weeks later, the average daily food intake of male rats stabilized at approximately 25~28 g per rat, while that of female rats remained stable at approximately 16~19 g per rat. The food utilization rate of all animals gradually decreased from the first week of the experiment. In the white blood cell (WBC) differential count results, significant differences were observed in the counts of WBCs, neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono) between males and females (P<0.001). However, there were no significant differences in the percentages of neutrophil (%Neut), lymphocyte (%Lymph), and monocyte (%Mono) between the sexes (P>0.05). The average red blood cell count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), platelet count (PLT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were higher in male animals than in female animals (P<0.05). The average values of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine phosphokinase (CK), lactate dehydrogenase (LDH), glucose (GLU), and triglyceride (TG) in male rats were higher than those in female rats (P<0.05). The urinary pH range for male animals was 5.0 to 8.5, while for female animals it was 6.5 to 9.0. The majority of male animals had a urinary specific gravity lower than 1.020, and the majority of female animals had a urinary specific gravity lower than 1.015. The weights of various organs (excluding the adrenal glands and reproductive organs) in male animals were heavier than those in female animals (P<0.001), while the organ/body weight ratios (excluding the kidneys and reproductive organs) of female animals were higher than those of male animals (P<0.001). ConclusionThis study summarizes the background reference ranges for body weight, food intake, hematology, and serum biochemistry indicators in SPF-grade SD rats in the untreated control group from six 90-day feeding trials conducted by the National Center for Safety Evaluation of Drugs. It provides important reference data for related research. By summarizing the background and spontaneous histopathological changes in rats, this study aids in the standardization and normalization of subsequent research, as well as in the evaluation and analysis of abnormal results.

OBJECTIVE: To assess the value of copy number variation sequencing (CNV-seq) for the diagnosis of children with intellectual disability (ID), developmental delay (DD), and autistic spectrum disorder (ASD). METHODS: Forty patients with ID/DD/ASD referred to Nanshan Maternity and Child Health Care Hospital from September 2018 to January 2022 were enrolled. G-banded karyotyping analysis was carried out for the patients. Genomic DNA was extracted from peripheral blood samples and subjected to CNV-Seq analysis to detect chromosome copy number variations (CNVs) in such patients. ClinVar, DECIPHER, OMIM and other database were searched for data annotation. RESULTS: Among the 40 patients (including 30 males and 10 females), 16, 15 and 6 were diagnosed with ID, DD and ASD, respectively. One patient had combined symptoms of ID and DD, whilst the remaining two had combined ID and ASD. Four patients were found with abnormal karyotypes, including 47,XY,+mar, 46,XY,inv(8)(p11.2q21.2), 46,XX,del(5)(p14) and 46,XX[76]/46,X,dup(X)(p21.1q12). Chromosome polymorphism was also found in two other patients. CNV-seq analysis has detected 32 CNVs in 20 patients (50.0%, 20/40). Pathogenic CNVs were found in 10 patients (25.0%), 15 CNVs of uncertain clinical significance were found in 12 patients (30.0%), and 7 likely benign CNVs were found in 4 patients (10.0%). CONCLUSION Chromosome CNVs play an important role in the pathogenesis of ID/DD/ASD. CNV-seq can detect chromosomal abnormalities including microdeletions and microduplications, which could provide a powerful tool for revealing the genetic etiology of ID/DD/ASD patients.
Pregnancy ; Child ; Male ; Humans ; Female ; DNA Copy Number Variations ; Intellectual Disability/genetics* ; Autism Spectrum Disorder/genetics* ; Developmental Disabilities/genetics* ; Abnormal Karyotype

Self-assembly is the basis of the formation of biological macromolecular structure. Enzyme-instructed self-assembly (EISA) with the help of tool enzymes, realizing the conversion of small molecular compounds to supramolecular nanostructures at specific sites, become a new strategy for drug discovery.In recent years, the exploration of EISA for developing malignant cancer therapy and imaging has made considerable progress, achieving the precise regulation and tumor targeting of nanostructures. This paper reviews the latest progress of EISA in the field of tumor diagnosis and treatment, the functions and characteristics of tool enzymes such as alkaline phosphatase, sirtuin, tyrosinase, γ-glutamyltranspeptidase and caspase-3,summarizes the research status of EISA targeting multiple organelles in tumor therapy, and introduces the application of EISA in tumor imaging, aiming to provide reference forthe research of EISA strategy in tumor diagnosis and treatment.

Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found that high glucose(HG)levels in the cerebrospinal fluid(CSF)of diabetic rats might enhance the effect of a subthreshold dose of the neurotoxin 6-hydroxydopamine(6-OHDA)on the development of motor disorders,and the damage to the nigrostriatal dopaminergic neuronal pathway.In vitro,HG promoted the 6-OHDA-induced apoptosis in PC12 cells differentiated to neurons with nerve growth factor(NGF)(NGF-PC12).Metabolomics showed that HG promoted hyperglycolysis in neurons and impaired tricarboxylic acid cycle(TCA cycle)activity,which was closely related to abnormal mito-chondrial fusion,thus resulting in mitochondrial loss.Interestingly,HG-induced upregulation of pyruvate kinase M2(PKM2)combined with 6-OHDA exposure not only mediated glycolysis but also promoted abnormal mitochondrial fusion by upregulating the expression of MFN2 in NGF-PC12 cells.In addition,we found that PKM2 knockdown rescued the abnormal mitochondrial fusion and cell apoptosis induced by HG+6-OHDA.Furthermore,we found that shikonin(SK),an inhibitor of PKM2,restored the mito-chondrial number,promoted TCA cycle activity,reversed hyperglycolysis,enhanced the tolerance of cultured neurons to 6-OHDA,and reduced the risk of PD in diabetic rats.Overall,our results indicate that diabetes promotes hyperglycolysis and abnormal mitochondrial fusion in neurons through the upre-gulation of PKM2,leading to an increase in the vulnerability of dopaminergic neurons to 6-OHDA.Thus,the inhibition of PKM2 and restoration of mitochondrial metabolic homeostasis/pathways may prevent the occurrence and development of diabetic PD.

Proteomic characterization of plasma is critical for the development of novel pharmacodynamic bio-markers.However,the vast dynamic range renders the profiling of proteomes extremely challenging.Here,we synthesized zeolite NaY and developed a simple and rapid method to achieve comprehensive and deep profiling of the plasma proteome using the plasma protein corona formed on zeolite NaY.Specifically,zeolite NaY and plasma were co-incubated to form plasma protein corona on zeolite NaY(NaY-PPC),followed by conventional protein identification using liquid chromatography-tandem mass spectrometry.NaY was able to significantly enhance the detection of low-abundance plasma proteins,minimizing the"masking"effect caused by high-abundance proteins.The relative abundance of middle-and low-abundance proteins increased substantially from 2.54％to 54.41％,and the top 20 high-abundance proteins decreased from 83.63％to 25.77％.Notably,our method can quantify approxi-mately 4000 plasma proteins with sensitivity up to pg/mL,compared to only about 600 proteins iden-tified from untreated plasma samples.A pilot study based on plasma samples from 30 lung adenocarcinoma patients and 15 healthy subjects demonstrated that our method could successfully distinguish between healthy and disease states.In summary,this work provides an advantageous tool for the exploration of plasma proteomics and its translational applications.

Objective To observe the therapeutic effect of probiotics combined with montelukast in children with Mycoplasma pneumoniae pneumonia(MPP)complicated with myocardial injury.Methods A total of 102 children with MPP and myocardial injury were selected as the study objects,and were randomly divided into control group and observation group,with 51 cases in each group.The control group was treated with montelukast,and the observation group was treated with probiotics based on the control group.The clinical efficacy,symptom resolution time,hospital stay,myocardial injury index,inflammatory cytokines,intestinal flora level and incidence of adverse reactions were compared between the two groups.Results The total effective rate of the observation group was 92.16％,which was significantly higher than 76.47％of the control group(P＜0.05).The disappearance time of symptoms of fever,cough,shortness of breath,chest tightness and chest pain and hospital stay in the observation group were significantly shorter than those in the control group(P＜0.05).After treat-ment,the levels of cardiac troponin T(cTnT),B-type natriuretic peptide(BNP),myoglobin(Mb)and cardiac fatty acid binding protein(H-FABP)in the observation group were significantly lower than those in the control group(P＜0.05);the levels of interleukin-1 receptor 1(IL-1R1),gamma-interferon(γ-IFN)and high mobility group protein 1(HMGB1)in the observation group were significantly lower than those inthe control group(P＜0.05);the number of Bifidobacteria,Lactobacillus aci-dophilus,Enterococcus faecalis and Bacillus cereus in the observation group were significantly higher than the control group,while the number of Escherichia coli and Salmonella were significantly lower than those in the control group(P＜0.05).There was no significant difference in the incidence of ad-verse reactions between the two groups(P＞0.05).Conclusion Probiotics combined with monte-lukast have good efficacy in MPP children with myocardial injury,which can shorten the time of symp-tom resolution and hospital stay,improve the indexes of myocardial injury in children,inhibit the lev-el of inflammatory cytokines,adjust the structure of intestinal flora disorder,and have good safety.

Objective To observe the therapeutic effect of probiotics combined with montelukast in children with Mycoplasma pneumoniae pneumonia(MPP)complicated with myocardial injury.Methods A total of 102 children with MPP and myocardial injury were selected as the study objects,and were randomly divided into control group and observation group,with 51 cases in each group.The control group was treated with montelukast,and the observation group was treated with probiotics based on the control group.The clinical efficacy,symptom resolution time,hospital stay,myocardial injury index,inflammatory cytokines,intestinal flora level and incidence of adverse reactions were compared between the two groups.Results The total effective rate of the observation group was 92.16％,which was significantly higher than 76.47％of the control group(P＜0.05).The disappearance time of symptoms of fever,cough,shortness of breath,chest tightness and chest pain and hospital stay in the observation group were significantly shorter than those in the control group(P＜0.05).After treat-ment,the levels of cardiac troponin T(cTnT),B-type natriuretic peptide(BNP),myoglobin(Mb)and cardiac fatty acid binding protein(H-FABP)in the observation group were significantly lower than those in the control group(P＜0.05);the levels of interleukin-1 receptor 1(IL-1R1),gamma-interferon(γ-IFN)and high mobility group protein 1(HMGB1)in the observation group were significantly lower than those inthe control group(P＜0.05);the number of Bifidobacteria,Lactobacillus aci-dophilus,Enterococcus faecalis and Bacillus cereus in the observation group were significantly higher than the control group,while the number of Escherichia coli and Salmonella were significantly lower than those in the control group(P＜0.05).There was no significant difference in the incidence of ad-verse reactions between the two groups(P＞0.05).Conclusion Probiotics combined with monte-lukast have good efficacy in MPP children with myocardial injury,which can shorten the time of symp-tom resolution and hospital stay,improve the indexes of myocardial injury in children,inhibit the lev-el of inflammatory cytokines,adjust the structure of intestinal flora disorder,and have good safety.

Objective To analyze the expression of KCNJ11 mRNA in gliomas and its prognostic value. Methods The clinical, histopathological and molecular pathological features of 273 patients with gliomas were collected from CGGA. We analyzed the differences of KCNJ11 mRNA expression in different types of gliomas and the survival time of patients with high and low expression of KCNJ11 mRNA in different subtypes of gliomas. Results The expression levels of KCNJ11 mRNA in young glioma and primary glioma patients were higher than those in old glioma and recurrent glioma patients, respectively (P=0.008, 0.001). The expression of KCNJ11 mRNA in oligodendroglioma was the highest, astrocytoma was the second, and glioblastoma was the lowest (P=0.000). The expression of KCNJ11 mRNA in WHOⅡ grade glioma was the highest, WHOⅢ was the second, and WHOⅣ was the lowest (P=0.000). The expression levels of KCNJ11 mRNA in IDH-mutant type glioma patients were higher than those in IDH-wild type glioma patients (P=0.000). The expression of KCNJ11 mRNA in deletion of 1p/19q glioma patients was higher than that in non-deletion of 1p/19q ones (P=0.000). The expression of KCNJ11 mRNA in MGMT methylated glioma patients was higher than that in non-methylated ones (P=0.036). The survival time of patients with high expression of KCNJ11 mRNA (≥2.77) was longer than that with low expression (P=0.000). Multivariate Cox analysis showed that the high expression of KCNJ11 mRNA was an independent factor affecting the good prognosis of patients with glioma. Conclusion The expression of KCNJ11 mRNA is negatively related to the malignant degree of the tumor. The high expression of KCNJ11 mRNA is an independent factor affecting the good prognosis of patients with glioma.

Objective:To investigate the mRNA expression of adenosine triphosphate binding cassette subfamily C member 8 (ABCC8) in gliomas and its application value as an index for predicting the prognosis of gliomas.Methods:The data of sex, age, histopathological type, WHO grade, isocitrate dehydrogenase (IDH) mutation, 1p/19q deletion, survival time and ABCC8 mRNA expression of 304 patients with gliomas (study group) in the Chinese glioma Genome Atlas (CGGA) database were collected. The prognostic factors of patients with gliomas were determined by univariate and multivariate Cox regression analysis. This result was verified by the data of 506 glioma patients in TCGA database and 271 glioma patients in REMBRANDT database.Results:The expression of ABCC8 mRNA in 304 cases of gliomas in the study group was 15.93±1.23. ABCC8 mRNA expression was correlated with age, histopathological type, WHO grade, IDH mutation status, 1p/19q deletion status and WHO molecular grade ( P<0.01). The median survival time of patients in ABCC8 mRNA high expression group was 85.60 months, which was significantly longer than 14.07 months in low expression group ( P<0.001). Univariate Cox regression analysis showed that age, histopathological type, WHO grade, IDH mutation status, 1p/19q deletion status, postoperative radiotherapy, postoperative chemotherapy and ABCC8 mRNA expression were related to the overall survival time of patients ( P<0.05). Multivariate Cox regression analysis showed that WHO grade ( HR=3.117, 95% CI: 1.970-4.930), 1p/19q deletion status ( HR=0.285, 95% CI: 0.155-0.526), postoperative chemotherapy ( HR=0.680, 95% CI: 0.488-0.948) and ABCC8 mRNA expression ( HR=0.690, 95% CI: 0.491-0.971) were independent factors for the overall survival of patients. In the TCGA dataset, the median survival time of patients with high ABCC8 mRNA expression group was 133.70 months, which was longer than 19.60 months of patients with low ABCC8 mRNA expression group ( P<0.001). In Rembrandt dataset, the median survival time of patients with high ABCC8 mRNA expression group was 37.93 months, which was longer than 15.83 months of patients with low expression group ( P<0.001). Conclusions:The expression level of ABCC8 mRNA in glioma is related to the degree of malignancy. The higher degree of malignancy, the lower the expression. The expression level of ABCC8 mRNA is an independent factor for the overall survival of patients with gliomas. The overall survival of patients with high ABCC8 mRNA expression is longer than that of patients with low ABCC8 mRNA expression.

Objective:To investigate the mRNA expression of adenosine triphosphate binding cassette subfamily C member 8 (ABCC8) in gliomas and its application value as an index for predicting the prognosis of gliomas.Methods:The data of sex, age, histopathological type, WHO grade, isocitrate dehydrogenase (IDH) mutation, 1p/19q deletion, survival time and ABCC8 mRNA expression of 304 patients with gliomas (study group) in the Chinese glioma Genome Atlas (CGGA) database were collected. The prognostic factors of patients with gliomas were determined by univariate and multivariate Cox regression analysis. This result was verified by the data of 506 glioma patients in TCGA database and 271 glioma patients in REMBRANDT database.Results:The expression of ABCC8 mRNA in 304 cases of gliomas in the study group was 15.93±1.23. ABCC8 mRNA expression was correlated with age, histopathological type, WHO grade, IDH mutation status, 1p/19q deletion status and WHO molecular grade ( P<0.01). The median survival time of patients in ABCC8 mRNA high expression group was 85.60 months, which was significantly longer than 14.07 months in low expression group ( P<0.001). Univariate Cox regression analysis showed that age, histopathological type, WHO grade, IDH mutation status, 1p/19q deletion status, postoperative radiotherapy, postoperative chemotherapy and ABCC8 mRNA expression were related to the overall survival time of patients ( P<0.05). Multivariate Cox regression analysis showed that WHO grade ( HR=3.117, 95% CI: 1.970-4.930), 1p/19q deletion status ( HR=0.285, 95% CI: 0.155-0.526), postoperative chemotherapy ( HR=0.680, 95% CI: 0.488-0.948) and ABCC8 mRNA expression ( HR=0.690, 95% CI: 0.491-0.971) were independent factors for the overall survival of patients. In the TCGA dataset, the median survival time of patients with high ABCC8 mRNA expression group was 133.70 months, which was longer than 19.60 months of patients with low ABCC8 mRNA expression group ( P<0.001). In Rembrandt dataset, the median survival time of patients with high ABCC8 mRNA expression group was 37.93 months, which was longer than 15.83 months of patients with low expression group ( P<0.001). Conclusions:The expression level of ABCC8 mRNA in glioma is related to the degree of malignancy. The higher degree of malignancy, the lower the expression. The expression level of ABCC8 mRNA is an independent factor for the overall survival of patients with gliomas. The overall survival of patients with high ABCC8 mRNA expression is longer than that of patients with low ABCC8 mRNA expression.