Objective To observe the effect of m⁶A methylation regulation on Notch1 pathway on the homing of BMSCs in asthma, and the intervention study of traditional Chinese medicine compound Yanghe Pingchuan Granules. Methods Rat bone mesenchymal stem cells(BMSC)and bronchial epithelial cells were cocultured. The extracted cells were divided into: bronchial epithelial cell group, asthma bronchial epithelial cell+mesenchymal stem cell co-culture group (co-culture group), co-culture cell+normal serum group, coculture cell+serum containing optimal drug group, siRNA FTO+normal serum group, siRNA FTO-NC+normal serum group, and siRNA FTO+serum containing optimal drug group. The vitality and cell cycle changes of co-cultured cells were detected. The level and markers of homing BMSC were detected by immunofluorescence staining. The expression of Notch1 pathway related genes were detected by qRT-PCR. The expression of Notch1 pathway related proteins were detected by Western blot. Results Compared with bronchial epithelial cell group, the co-cultured cell group showed an increase in the homing level of BMSCs and the expression of C-X-C motif chemokine receptor 4 (CXCR4), stromal cell-derived factor 1 (SDF-1), Notch1, transcription factor recombination signal binding protein-J (RBP-J), and hairy enhancer of split 1 (Hes1) proteins. Compared with the co-cultured cell group and co-cultured cell+normal serum group, the co-cultured cell+serum containing optimal drug group showed an increase in the homing level of BMSCs and the expressions of CXCR4 and SDF-1, while the protein and mRNA levels of Notch1 and Hes1 decreased. Compared with the siRNA FTO-NC+normal serum group, the siRNA FTO+normal serum group showed an increase in the levels of Notch1, activated Notch1, RBP-J, Hes1 protein, and cell viability, while the level of homing BMSC decreased. Compared with siRNA FTO+normal serum group, the levels of Notch1, RBP-J mRNA, activated Notch1, and Hes1 protein decreased, while the level of homing BMSCs increased in siRNA FTO+serum containing optimal drug group. The levels of Notch1, RBP-J, and Hes1 mRNA were reduced in the co-cultured cells+serum containing optimal drug group. Compared with siRNA FTO+serum containing optimal drug group, the expressions of Notch1, activated Notch1, RBP-J, Hes1 protein and cell viability decreased, while the level of homing BMSCs increased in the co-cultured cells+serum containing optimal drug group. Conclusion Yanghe Pingchuan Granules may promote the homing of BMSCs in asthma and alleviate asthma inflammation by upregulating the expression of FTO and inhibiting the expression of downstream genes in the Notch1 signaling pathway.
Animals ; Receptor, Notch1/genetics* ; Mesenchymal Stem Cells/cytology* ; Asthma/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Signal Transduction/drug effects* ; Rats ; Coculture Techniques ; Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics* ; Epithelial Cells/metabolism* ; Rats, Sprague-Dawley ; Cells, Cultured ; Male

[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

Objective To observe the clinical and CT features of neonatal adrenal cystic neuroblastoma(CNB).Methods Eight newborns with adrenal CNB confirmed by surgical pathology were retrospectively analyzed.The clinical data were recorded,and the plain and enhancement abdominal CT manifestations were observed.Results Among 8 cases,6(6/8,75.00％)were detected with prenatal ultrasound,while 2(2/8,25.00％)were detected after birth with ultrasound,all with single adrenal grand lesion,located half in left and half in right adrenal gland(each 4/8,50.00％).The maximum diameter of CNB lesion was 2.3-6.1 cm,with the median maximum diameter of 4.5 cm.CT showed all 8 lesions(8/8,100％)presented as single localized adrenal grand thick-walled cystic lesion,among which 3(3/8,37.50％)with uniform density within the cysts,3(3/8,37.50％)with internal septum within the cysts,1(1/8,12.50％)with slight floating debris and the rest 1(1/8,12.50％)with both internal septum and floating debris in the cyst.No calcification,cross the midline nor surround blood vessels were observed.Seven(7/8,87.50％)lesions had clear while 1(1/8,12.50％)had unclear boundaries,all mildly compressed surrounding structures.The capsule wall and internal septum of the cysts slightly enhanced after enhancement.Multiple liver metastases occurred in 2 cases.Conclusion Most neonatal adrenal CNB were detected before delivery,which mainly presented as thick-walled cystic mass with clear boundary,accompanied by septa and floating debris,and the cystic wall and septa slightly enhanced after enhancement,and liver metastasis might occur.

Hepatocellular carcinoma (HCC) is a common malignant tumor with poor prognosis and high mortality. In this study, we demonstrated a novel vaccine targeting HCC and tumor neovascular endothelial cells by fusing recombinant MHCC97H cells expressing porcine α-1,3-galactose epitopes (αGal) and endorphin extracellular domains (END) with dendritic cells (DCs) from healthy volunteers. END⁺/Gal⁺-MHCC97H/DC fusion cells induced cytotoxic T lymphocytes (CTLs) and secretion of interferon-gamma (IFN-γ). CTLs targeted cells expressing αGal and END and tumor angiogenesis. The fused cell vaccine can effectively inhibit tumor growth and prolong the survival time of human hepatoma mice, indicating the high clinical potential of this new cell based vaccine.

OBJECTIVE: To analyze the characteristics of skeletal muscle cells gene markers in septic patients by using bioinformatics. METHODS: The differential gene expression of marker microarrays (GSE13205) in skeletal muscle tissue of patients with sepsis was obtained from gene expression omnibus (GEO) database of National Center for Biotechnology Information (NCBI). Gene differential expression analysis was carried out using online GEO2R provided by NCBI. Data processing, analysis and mapping were carried out using online bioinformatics array research tool (BART) and Cytoscpe software, the software of the national resource for network biology. Functional annotation and pathway analysis of differential expression genes were performed using Kyoto encyclopedia of genes and genomes (KEGG) and gene ontology (GO) provided by the database for annotation, visualization and integrated discovery (DAVID), and protein interaction analysis was further performed in search tool for the retrieve of interacting genes/proteins (STRING-DB). RESULTS: The TOP250 genes were extracted from the GSE13205 dataset. A total of 242 differentially expressed genes were included in the analysis. Among them, 78 up-regulated genes and 164 down-regulated genes were identified. After extensive data analysis, these differentially expressed genes were enriched into different biological processes or subsets of molecular functions, mainly enriched in the positive and negative regulation of growth, mineral absorption and other pathways. The 14 most closely related genes among differentially expressed genes were identified from the protein interaction network. CONCLUSIONS The differential expression genes in patients with sepsis were mainly concentrated on cell growth and apoptosis and mediating tumor-related immune function regulation.
Computational Biology ; Gene Expression Profiling ; Gene Ontology ; Gene Regulatory Networks ; Genetic Markers ; Humans ; Muscle, Skeletal/metabolism* ; Sepsis/genetics*

Objective To evaluate the expression and clinical significance of plasma circulating microRNA-23b in patients with myasthenia gravis (MG).Methods Plasma samples from 32 MG patients and 32 healthy people were collected in the Second Affiliated Hospital of Harbin Medical University from October 2015 to August 2016. Expression of microRNA-23b was detected by quantitative reverse transcription polymerase chain reaction .The relationship among microRNA-23b expression level, clinical features and quantitative myasthenia gravis score ( QMG ) was analyzed. Results The expression of circulating microRNA-23b in plasma was significantly higher in MG patients (4.587 (3.654, 5.423)) compared with control group (2.889 (1.968, 4.027), Z=-4.169, P<0.01), meanwhile it was positively related to QMG (r=0.661,P<0.01).But the expression of circulating microRNA-23b had no statistically significant difference in MG subgroups .Conclusion Plasma circulating microRNA-23b is highly expressed in MG and has positive correlation with MG severity .

Objective To study whether crystal-8 exerts analgesic effect on the central nervous system,and to observe the relationship between crystal-8 and receptors. Methods A total of 32 SD rats were randomly divided into the 0.9% sodium chloride solution group, positive control group ( rotundine 2 mg·kg-1 ) , high dose of crystal-8 group ( 2 mg·kg-1 ) and low dose of crystal-8 group ( 1 mg · kg-1 ) . The changes of pain threshold were measured by the thermal stimulation test following intracerebroventricular (ICV) injection.The SD rats or mice were randomly divided into the 0.9% sodium chloride solution group, receptor tool medicine group ( including naloxone, reserpine, haloperidol, scopolamine) , crystal-8 group, receptor tool medicine plus crystal-8 group. The pain threshold was detected by the thermal stimulation test at 15, 30, 45, 60, 90, 120 min after dosing, then the influence of analgesic effect of crystal-8 on the neurotransmitter was observed. Results Compared with the 0.9% sodium chloride solution group, the pain threshold of rats was improved after taking the crystal-8 by intracerebroventricular (ICV) injection(P<0.01).Compared with the crystal-8 group, the naloxone plus crystal-8 group, the haloperidol plus crystal-8 group, the scopolamine plus crystal-8 group couldn't increase the pain threshold, there was no significant difference among these groups(P>0.05).However, Reserpine plus crystal-8 group could significantly decrease the pain threshold in rats compared with the crystal-8 group(P<0.01).The analgesic effect of crystal-8 was interfered by reserpine. Conclusion The analgesic effect of crystal-8 may be involved in the central mechanism,which relates to monoamine neurotransmitters, but has nothing to do with the opioid receptor, M receptor, and the inhibition of central DA receptor.

Objective To investigate the expression level of thymus microRNA-27a-3p in patients with myasthenia gravis (MG) and to explore the pathogenesis of MG.Methods Thymus tissue samples from 36 cases were collected from December 2014 to February 2015 in the Second Affiliated Hospital of Harbin Medical University.Nineteen thymus tissue samples of MG group were collected from department of chest surgery,17 thymus tissue samples of control group were collected from department of chest surgery or congenital heart disease patients from department of cardiac surgery.The expression of microRNA-27a-3p in the thymus from 36 patients was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR),using U6 as housekeeping control.The Wilcoxon Rank-Sum test was used to analyze the relative expression of microRNA-27a-3p of the two groups.Spearman rank correlation was used to determine the correlation coefficient between microRNA-27a-3p and Quantitative Myasthenia Gravis Score (QMGS).Results (1) The expression level of microRNA-27a-3p in thymus was significantly higher in MG group (0.195(0.049,0.714)) compared with control group (0.045(0.004,0.088),Z =-2.646,P =0.008).(2) Nineteen MG patients were included in the study,out of which 7 were ocular myasthenia gravis (OMG) patients and 12 were generalized myasthenia gravis (GMG) patients.The expression of microRNA-27a-3p in GMG patients (0.493 (0.157,1.123)) was significantly higher than that in OMG patients (0.035 (0.008,0.103),Z =-2.620,P =0.009).(3) There was a positive correlation between the expression of microRNA-27a-3p and QMGS (r =0.576,P =0.010).Conclusions The expression of microRNA-27a-3p in thymus is significantly up-regulated in the patients with MG.MicroRNA-27a-3p may be associated with MG severity and significantly elevated in GMG patients compared with OMG patients.

Objective To investigate the association between myasthenia gravis (MG) and single nucleotide polymorphisms (SNPs) of PTPN22 + 1858C/T,CTLA-4 (+ 49A/G;-1772C/T;-1661A/G),KRAS(rs9226),BCL2(rs4987855) and IGF-1R(rs34804698) genes.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to detect the gene types of SNPs in 76 MG patients who were enrolled in the Second Affiliated Hospital of Harbin Medical University from July 2011 to June 2015 and 59 healthy blood donors.Results In MG patients,the frequences of CTLA-4 +49A/G(rs231775) (57.9％) and-1772C/T (rs733618) (43.4％) were higher than that in the healthy controls (22.1％) (x2 =35.252,P =0.000; x2 =4.098,P =0.043).The frequence of CTLA-4 +49A/G in MG patients combined with thymoma (25.6％) was higher than other subgroups (thymic hyperplasia group:13.8％; normal thymus group:18.4％)(x2 =7.564,P=0.006; x2 =7.155,P=0.007).Meanwhile,the frequence of the C-1772 allele was higher in thymoma group (19.7％) compared with other two groups (thymic hyperplasia group:9.86％ ; normal thymus group:13.8％) (x2 =5.331,P =0.021 ;x2 =4.411,P =0.036).However,the other SNPs were not associated with the risk of MG.Conclusion There are associations of MG with CTLA-4 + 49A/G and-1772C/T SNPs,but not with PTPN,KRAS,BC12 and IGF-1R SNPs.

Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.
Amplified Fragment Length Polymorphism Analysis ; methods ; Cytoplasm ; metabolism ; DNA, Mitochondrial ; genetics ; DNA, Plant ; genetics ; Gene Expression Profiling ; Genotype ; Plant Infertility ; genetics ; Triticum ; genetics