Objective·To investigate the differential gene expression of microglia in the gray and white matter of multiple sclerosis(MS)using single-nucleus transcriptomic analysis,aiming to explore their roles in disease progression,and identify key transcriptional regulatory networks associated with the disease.Methods·snRNA-seq data of frozen human brain tissue samples from MS patients and control individuals were obtained from the Gene Expression Omnibus(GEO)database.R language,along with R packages such as Seurat,was employed to identify cell types based on specific cell markers.Microglia were extracted from the identified cell populations and classified based on their anatomical origin,either gray matter or white matter.Dimensionality reduction and clustering techniques were utilized to identify distinct microglial subpopulations with differential characteristics.Differentially expressed genes(DEGs)between the MS and control groups at the subpopulation level were analyzed by using the Seurat package.Gene set enrichment analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)was conducted on the DEGs to further explore the biological significance of these differences.Monocle3 was used for pseudotime analysis to study dynamic changes in microglia subpopulations during disease progression.Single cell regulatory network inference and clustering(SCENIC)method was applied to analyze transcription factor(TF)regulatory networks,aiming to identify key transcription factors potentially involved in MS regulation.Results·After quality control,a total of 149 062 nuclei were retained for analysis.Following dimensional reduction and clustering,12 238 microglia were identified by using key markers,including DOCK8,CSF1R,P2RY12,and CD74.The results of GO and KEGG pathway analysis showed that in gray matter microglia,functions such as endocytosis,ion homeostasis,and lipid localization were downregulated during disease progression,while in white matter microglia,functions such as protein folding,cytoplasmic translation,and response to thermal stimuli were upregulated.SCENIC analysis revealed that the expression of transcription factors such as FLI1,MITF,and FOXP1 was upregulated in MS.Conclusion·Microglia play a critical role in MS,with white matter microglia being more significantly impacted by MS than their gray matter counterparts.Transcription factors such as FLI1,MITF,and FOXP1 are identified as key regulators involved in disease modulation,with their associated transcriptional regulatory networks playing a central role in disease modulation.

Objective·To investigate the differential gene expression of microglia in the gray and white matter of multiple sclerosis(MS)using single-nucleus transcriptomic analysis,aiming to explore their roles in disease progression,and identify key transcriptional regulatory networks associated with the disease.Methods·snRNA-seq data of frozen human brain tissue samples from MS patients and control individuals were obtained from the Gene Expression Omnibus(GEO)database.R language,along with R packages such as Seurat,was employed to identify cell types based on specific cell markers.Microglia were extracted from the identified cell populations and classified based on their anatomical origin,either gray matter or white matter.Dimensionality reduction and clustering techniques were utilized to identify distinct microglial subpopulations with differential characteristics.Differentially expressed genes(DEGs)between the MS and control groups at the subpopulation level were analyzed by using the Seurat package.Gene set enrichment analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)was conducted on the DEGs to further explore the biological significance of these differences.Monocle3 was used for pseudotime analysis to study dynamic changes in microglia subpopulations during disease progression.Single cell regulatory network inference and clustering(SCENIC)method was applied to analyze transcription factor(TF)regulatory networks,aiming to identify key transcription factors potentially involved in MS regulation.Results·After quality control,a total of 149 062 nuclei were retained for analysis.Following dimensional reduction and clustering,12 238 microglia were identified by using key markers,including DOCK8,CSF1R,P2RY12,and CD74.The results of GO and KEGG pathway analysis showed that in gray matter microglia,functions such as endocytosis,ion homeostasis,and lipid localization were downregulated during disease progression,while in white matter microglia,functions such as protein folding,cytoplasmic translation,and response to thermal stimuli were upregulated.SCENIC analysis revealed that the expression of transcription factors such as FLI1,MITF,and FOXP1 was upregulated in MS.Conclusion·Microglia play a critical role in MS,with white matter microglia being more significantly impacted by MS than their gray matter counterparts.Transcription factors such as FLI1,MITF,and FOXP1 are identified as key regulators involved in disease modulation,with their associated transcriptional regulatory networks playing a central role in disease modulation.

Objective:To investigate the application value of digital X-ray radiotherapy and single photon emission computed tomography(SPECT)radionuclide bone imaging in the diagnosis of intertrochanteric fracture of femur.Methods:A total of 70 patients who were diagnosed as intertrochanteric fracture of femur in Shijiazhuang People's hospital from January 2017 to December 2020 were selected,and they were divided into study group and control group according to random number method,with 35 cases in each group.Digital X-ray radiography combined with SPECT radionuclide bone imaging was used in the study group,and digital X-ray radiography was used in the control group.And then,the accuracy of the diagnostic results between the two groups were compared.Results:The radiographic examination indicated that 31 cases(88.6%)of 35 cases in the study group were confirmed,and 24 cases(68.6%)of 35 cases in the control group were confirmed.The diagnostic accuracy rate of the study group was significantly higher than that of the control group,and the difference was statistically significant(x2=4.158,P<0.05).The preoperative imaging diagnosis indicated that diagnosed number of fracture blocks in the study group was(3.57±0.50),which was significantly higher than that(2.67±0.40)in the control group,and the difference of that between two groups was statistically significant(t=8.315,P<0.05).In the actual intraoperative examining for bone continuity,5 cases were continuity and 30 cases were non-continuity.In the bone stability,13 cases were stability and 22 cases were non-stability.The study group was closer to intraoperative bone continuity and bone stability,and the differences of them between two groups were statistically significant(x2=12.857,4.644,P<0.05),respectively.Conclusion:The combination of digital X-ray radiography and SPECT radionuclide bone imaging has higher diagnostic value for intertrochanteric fracture of femur,which can provide important reference for clinical diagnosis.

ObjectiveTo observe the effects of Coptidis Rhizoma-Fermentum Rubrum on non-alcoholic fatty liver disease （NAFLD） in mice and explore its possible mechanisms. MethodSixty male SPF C57BL/6J mice were randomly divided into six groups： control group， model group， low-， medium-， and high-dose Coptidis Rhizoma-Fermentum Rubrum group （0.75， 1.5， 3 g·kg^-1）， and metformin group （0.075 g·kg^-1）， with 10 mice in each group. NAFLD mouse models were induced by high-fat diet feeding for 24 weeks. The low， medium， and high-dose Coptidis Rhizoma-Fermentum Rubrum groups were administered corresponding doses of Coptidis Rhizoma-Fermentum Rubrum by gavage， while the control and model groups received an equivalent amount of saline for four weeks. Serum total cholesterol （TC）， triglycerides （TG）， free fatty acids （FFA）， and liver function markers including alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were measured using an automatic biochemical analyzer. Hematoxylin-eosin （HE） and oil red O staining were used to detect liver lipid deposition， and Prussian blue staining was used to measure liver ferrous ion levels. Western blot was performed to detect the expression of key proteins in the nuclear factor erythroid-2-related factor 2 （Nrf2）-glutathione peroxidase 4 （GPX4） axis. ResultAfter 24 weeks of high-fat feeding， compared with the control group， the model group showed significant increases in body weight， liver weight and liver index， and serum lipid levels （P<0.01）， as well as substantial hepatic lipid deposition with marked steatosis. Compared with the model group， Coptidis Rhizoma-Fermentum Rubrum intervention reduced body weight （P<0.01）， liver weight and liver index （P<0.01）， and serum lipid levels （P<0.05， P<0.01）， improved liver function （P<0.01）， and decreased hepatic lipid deposition， with the low-dose Coptidis Rhizoma-Fermentum Rubrum group showing the best effect. Western blot results showed that compared with those in the control group， the expression levels of Nrf2， heme oxygenase-1 （HO-1）， kelch-like ECH-associated protein 1（Keap1）， and GPX4 proteins in the model group were decreased （P<0.05， P<0.01）. Compared with the model group， Coptidis Rhizoma-Fermentum Rubrum increased the expression levels of these proteins （P<0.05， P<0.01）. ConclusionCoptidis Rhizoma-Fermentum Rubrum can alleviate fatty liver in mice， improve liver function， and reduce hepatic lipid deposition， possibly by regulating the Nrf2/GPX4 ferroptosis axis.

Objective:To evaluate the efficacy and safety of sodium hyaluronate gel in preventing adhesion after prophylactic enterostomy.Methods:One hundred and twenty four patients from 6 hospitals were enrolled in this prospective multi-center randomized controlled trial. Patients were randomized into the study group ( n=59) or the control group ( n=65).All patients underwent prophylactic enterostomy. Patients of study group received odium hyaluronate gel for adhesion-prevention,while those in control group did not receive any adhesion-prevention treatment. The incidence of moderate to severe adhesion around the incision in the stoma area were evalutated during stoma reduction surgery. Results:The incidence of moderate to severe adhesion around the incision in the stoma area was 6.3% in the study group, the difference was statistically significant ( P<0.05) compared to that of the control group (32.6%). Conclusion:Sodium hyaluronate gel can safely and effectively reduce the incidence of moderate and severe adhesions after abdominal surgery.

Objective To construct DNA vaccine expressing Mycobacterium tuberculosis(Mtb) immunodominant antigen Ag85A and analyze its anti-tuberculosis T cell responses in BCG primed-mice after DNA vaccination boosting.Methods The coding gene of Ag85A mature fragment was amplified by PCR with H37Rv genomic DNA as template,and then cloned into the eukaryotic expression vector pVAX1 to construct Ag85A DNA vaccine.After purification,Ag85A DNA vaccine was injected intramuscularly twice in BCG primed-mice with BCG vaccination and DNA vaccination alone as control.Eight weeks post-vaccination,spleen lymphocytes were separated and were then used to analyze Mtb antigen specific effector T cell response and polyfuntional IFN-γ/TNF-α/IL-2 secreting CD4+ T cell frequencies and intensities,and CD8+T cell responses by IFN-γ ELISPOT assay and intracellular staining,respectively.Results Compared to BCG vaccinated-and DNA vaccinated-mice,Ag85A DNA boosting not only enhanced significantly BCG primed-mice IFN-γ+TNF-α+IL-2+,IFN-γ+ IL-2+,TNF-α+IL-2+ and IL-2+ CD4+ T cell frequencies and IL-2 secretion,but also improved significantly IFN-γ-secreting and IL-2-secreting CD8+ T cell frequencies.Condusion Ag85A DNA vaccine was constructed successfully and was demonstrated to enhance significantly BCG primed-mice Mtb antigen specific CD4+ and CD8+ T cell responses when boosting,which is beneficial to improve BCG immunogenicity and its waning immune protection against Mtb.

Dendritic cells are the most powerful cells to induce T cell immune reaction. DC derived from leukemia also possess the ability of the antigen processing and presentation. DC pulsed with tumor antigen can effectively initiate T cell immune reaction and induce T cell to cytotoxic T lymphocytes in vitro and in vivo. In this text, we introduce the advanced development of the biological character of DC, tumor immunology, the mechanism of tumor cell escaping immune surveillance and DC in leukemia immunotherapy.

Objective:Clonidine,by activating peripheral α-sbrenoceptors, produces transient pressor response after i.v.injection in anesthetized animals.Moxonidine, with at least 40-fold higher affinity to I1-imidazoline receptors than to α2-adrenoceptors,produces also a transient pressor response. This work was designed to investigate whether I1-imidazoline receptors are involved in this pressor effect of moxonidine. Methods:Female spontaneously hypertensive rats(SHRs,aged 14-16 weeks)were anesthetized with urethane.To observe the transient pressor responses,moxonidine 0.1,0.3,1.0mg/kg(intravenous,i.v),2.0μg(intracerebroventricular,i.c.v.)and 1.0,10.0mg/kg(intragastric,i.g.)were administrated in different groups of rats.To evaluate the roles of α1-adrenoceptors,α2-adrenoceptors and I1-imidazoline receptors in the transient pressor responses to moxonidine, prazosin(10.0μg/kg),yohimbine(2.0mg/kg),phentolamine(0.2mg/kg),idazoxan(1.0mg/kg)or yohimbine+idazoxan(2.0mg/kg+1.0mg/kg)were intravenously given to the animals before moxonidine 0.3mg/kg (i.v.).Results:It was found that i.v.moxonidine produced a greater pressor response than clonidine when producing a similar reduction of blood pressure.This effect of moxonidine was not influenced by prazosin, but was partly inhibited by yohimbine, phentolamine or idazoxan,and completely blocked by the combination of yohimbine and idzaxon.Neither i.c.v.injection nor i.g. administration of moxonidine induced transient pressor responses.Conclusion:The transient pressor response of i.v. moxonidine is mediated by both peripheral I1-imidazoline receptors and α2-adrenoceptors.

Objective To explore the epidemiological features of the metabolic syndrome in a cohort study of Xiangtan Mengkuang in China.Methods The prevalence rate,means and standard deviation of various risk factors associated with metabolic syndrome were analysed on the basis of base-line survey of risk factors of 5792 subjects(aged 25 years-old～) in 2004.Results (1)The prevalence rate of metabolic syndrome was 12.9%(12.4% in males,and 13.7% in females,P

This study aims at exploring the effects of tensile strain and loading time on the secretion of IL-1 beta of human periodontal ligament fibroblast. Five tensile strain values including 0%, 8%, 12%, 16%, 20% and three loading time including 24 h, 48 h, 72 h are set in this study. The prepared cell samples are mounted on the self-devised loading apparatus in vitro. The content of IL-1 beta in each sample was determined using double-antibody ELISA. The secretion amount of IL-1 beta per day is directly proportional to the loading time and tensile strain value in tensile strain group of 8%, 12%, 16%. The secretion amount of IL-1 beta reaches its maximum at tensile strain value of 16% in loading time groups. Loaded strain for 24 h and 48 h, the secretion amount of IL-1 beta at tensile strain value of 20% is obviously more than that at value of 0%, but the amount already begin to decrease apparently. Loaded strain for 72 h, the secretion amount of IL-1 beta decreases to a great extent that it is less than the amount at strain value of 0%. The tensile strain stimulates the secretion of IL-1 beta by human periodontal ligament fibroblast when the strain is under normal physiological extent, but the stimulation effect fades out as time goes on.
Cells, Cultured ; Fibroblasts ; secretion ; Humans ; Interleukin-1 ; secretion ; Periodontal Ligament ; cytology ; secretion ; Tensile Strength ; Time Factors