ObjectiveTo investigate the heterogeneity and transcriptomic characteristics of T-cell subsets in the liver of mice with nonalcoholic steatohepatitis （NASH） at the single-cell level using single-cell RNA sequencing （scRNA-seq）， and to provide a reference for studying the mechanism of action of T cells in NASH. MethodsSix male C57BL/6 mice were randomly divided into control group fed with regular diet and NASH group fed with methionine-choline-deficient （MCD） diet， with three mice in each group， and liver tissue was collected for scRNA-seq after 6 weeks of modeling. Specific differentially expressed genes were analyzed between T-cell subsets， and related analyses were performed， including dimension clustering， cell type annotation， t-distributed stochastic neighbor embedding （t-SNE）， violin plot， gene ontology （GO） functional enrichment analysis， and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. Immunofluorescent staining was used to observe the expression of the T cell marker Tcrα and the specific marker genes Tcf7 and Cxcr6 in the liver of mice in the two groups. The independent-samples t test was used for comparison of continuous data between two groups. ResultsTwo T cell subsets were identified in the liver of mice， and the percentage of cluster 6 decreased from 58.5% in the control group to 48.7% in the NASH group. The top four specific genes were Nsg2， Cd8b1， Cd8a， and Tcf7. Tcf7， a characteristic marker gene for cluster 6， was expressed in 65% of cells in cluster 6， and therefore， cluster 6 was defined as Tcf7⁺ T cells. The GO and KEGG enrichment analyses showed that the differentially expressed genes of cluster 6 were involved in T cell activation， leukocyte adhesion， binding ubiquitin-like protein ligase， and the signaling pathways for Th17， Th1， and Th2 cell differentiation. The percentage of cluster 7 increased from 41.5% in the control group to 51.3% in the NASH group. The top four specific genes of cluster 7 were Cd40lg， Tcrg-C1， Il2rα， and Cxcr6. Cxcr6 was expressed in 90% of cells in cluster 7， and therefore， cluster 7 was defined as Cxcr6⁺ T cells. The GO and KEGG enrichment analyses showed that cluster 7 was involved in T cell activation， cytokine production， the T cell receptor signaling pathway， and the Th17 cell differentiation and MAPK signaling pathway. Immunofluorescence assay showed that compared with the control group， the NASH group showed a significant reduction in the area with positive co-expression of Tcf7 protein and Tcrα protein （1.80%±0.67% vs 0.33%±0.13%， P<0.05） and a significant increase in the area with positive co-expression of Cxcr6 protein and Tcrα protein （0.50%±0.09% vs 2.66%± 0.33%， P<0.001）. ConclusionThere is a reduction in the percentage of Tcf7⁺ T cells and an increase in the percentage of Cxcr6⁺ T cells in NASH mice， revealing the characteristics and differences of T cells in the liver of NASH mice.

The research focus of diabetes, a common chronic metabolic disease, has shifted from individual factors to environmental factors at the population level. Epidemiological studies suggest an association between exposure to environmental pollutants and the risk of diabetes. Major environmental pollutants include organochlorine pesticides, polychlorinated biphenyls, phthalates and their metabolites, and arsenics, which primarily enter the human body through the skin, respiratory tract, and digestive system. Long-term exposure to these pollutants can affect the pathology of diabetes through various mechanisms, such as promoting insulin resistance, causing insulin secretion deficiencies, inducing oxidative stress-induced glucose metabolism disorders, and affecting DNA methylation. However, the potential damaging mechanism of the impact of environmental pollutants on diabetes remain unclear. Limitations such as insufficient sample sizes, uncertainties regarding exposure time and dosage, and differences between single and co-exposures. In the future, it is necessary to focus on exploring and analyzing the mechanisms of environmental pollutant exposure on diabetes to develop effective prevention strategies, control and reduce the incidence and development of diabetes, and provide new insights into its diagnosis and treatment.

By organizing and studying on the original works of LI Wenrong， this paper aimed to explore his theories and methods for treating liver diseases. LI Wenrong believes that the liver is easily prone to physiological disturbances， difficult to pacify， and can be compared to a “chariot” that moves horizontally and vertically， acting as a “general” for both civil and military affairs. Pathologically， liver diseases tend to spread to other organs， resulting in ever-changing pathological changes， often affecting emotional health. In terms of therapeutic methods， LI Wenrong proposes ten methods including draining fire to calm the liver， nourishing water to moisten wood， clearing metal to calm the liver， calming gallbladder and harmonizing liver， subduing yang with shell-drug， treating liver diseases by supplementing spleen， soothing the liver with sweetness， astringing the liver with sourness， relieving the liver with spiciness， and draining the liver with excess-fire. These methods have great guidance and reference value for contemporary clinical practice in traditional Chinese medicine.

Children and adults differ significantly in physiology,biochemistry and immune function,which leads to sig-nificant differences in blood transfusion strategies between children and adults.To guide the clinical transfusion practice of pediatric patients and improve the prognosis of children,the National Health Commission organized the formulation and re-lease of the health industry standard Guideline for Pediatric Transfusion(WS/T 795-2022).This paper will briefly introduce some concepts that help understand of the Standard and the preparation process of the Standard,and explain and interpret the preparation of the"scope","general provisions"and"factors to consider"of the Standard,hoping to contribute to the understanding and implementation of the Standard.

Objective:To study the complete genome characterization of Human Astrovirus (HAstV) in Shandong Province.Methods:Stool samples from acute flaccid paralysis (AFP) surveillance in Shandong Province from 2020 to 2022 were collected, and HAstV nucleic acid was examined by real-time quantitative PCR (qPCR). Next-generation sequencing (NGS) was conducted for the positive samples to obtain complete genome sequences and identify the genotype. Homology comparison and phylogenetic analysis were performed by using BioEdit and Mega software.Results:A total of 667 samples were examined by qPCR, of which 14 were HAstV-positive (2.1%), including HAstV-1 ( n=6), MLB1 ( n=6), MLB2 ( n=1), and VA2 ( n=1). The complete genome sequences were obtained from 11 samples. The six HAstV-1 sequences of this study had 98.2% to 99.9% nt similarities with each other and 87.6% to 98.6% with those from other regions. The four MLB1 sequences of this study had 99.1% to 99.9% nt similarities with each other and 92.2% to 99.4% with those from other regions. The VA2 sequence of this study had 96.0% to 96.3% nt similarities with those from other regions. Phylogenetic analysis based on ORF2 region showed that the local HAstV-1 sequences were most closely related to Japanese strains, and had distinct topology with phylogenies based on ORF1a and ORF1b regions. Conclusion:The complete genome sequences of 11 HAstV strains are obtained, and the VA2 complete genome is found.

Objective To analyze the seroepidemiology characteristics of Epstein-Barr virus(EBV)infection in children in Shenyang.Methods From June 2022 to May 2023,serum was collected from 12 083 children at Shengjing hospital of China Medical University.Serum capsid antigen(VCA)-IgM,VCA-IgG,EBV nuclear antigen-IgG(EBNA-IgG),and early antigen-IgG(EA-IgG)antibodies were detected using the LIAISION chemiluminescence immunoassay.EBV-DNA was detected using real-time PCR.Differences in antibody positivity rates between sexes,ages,and seasons of onset were compared.Results In 12 083 patients,the positive rates of VCA-IgM,VCA-IgG,EBNA-IgG,and EA-IgG were 9.95％(1 202/12 083),50.57％(6 110/12 083),46.03％(5 562/12 083)and 4.93％(596/12 083).The positive rates of VCA-IgM and EA-IgG were lower in male patients(9.09％and 4.44％,respectively)than in female patients(11.10％,5.60％;all P＜0.05).The differences in the positive rates for VCA-IgM,VCA-IgG,EBNA-IgG,and EA-IgG antibodies in children of different ages were statistically significant(all P＜0.05).Differences in the positive rates of VCA-IgG,EBNA-IgG,and EA-IgG antibodies were statistically significant when compared between seasons(all P＜0.05).Fourteen EBV antibody-positive combinations were detected,of which the main combination was VCA-IgG and EBNA-IgG double-positive,with a total of 4 741 cases(39.24％).Of the 3 712 children who underwent EBV-DNA detection testing,3 034(81.73％)were EBV-DNA-negative and 678(18.27％)were EBV-DNA-posi-tive.VCA-IgG and EBNA-IgG double positivity was the most common in EBV-DNA-negative and EBV-DNA-positive children,accounting for 983(26.48％)and 194 cases(5.23％),respectively.Conclusion Both VCA-IgG and EBNA-IgG antibodies are main positive in children with EBV infection in Shenyang.The positive rate of EBV antibodies is lower in boys than in girls.The positive rates of EBV anti-bodies differ in children of different ages and seasons of onset.

Sepsis is a complex syndrome caused by multiple pathogens and involves multiple organ failure, particularly spleen dysfunction. In 2017, the worldwide incidence was 48.9 million sepsis cases and 11 million sepsis-related deaths were reported (Rudd et al., 2020). Inflammation, oxidative stress, and apoptosis are the most common pathologies seen in sepsis. Liensinine (LIE) is a bisbenzylisoquinoline-type alkaloid extracted from the seed embryo of Nelumbo nucifera. Lotus seed hearts have high content of LIE which mainly has antihypertensive and antiarrhythmic pharmacological effects. It can exert anti-carcinogenic activity by regulating cell, inflammation, and apoptosis signaling pathways (Manogaran et al., 2019). However, its protective effect from sepsis-induced spleen damage is unknown. In this research, we established a mouse sepsis model induced by lipopolysaccharide (LPS) and investigated the protective effects of LIE on sepsis spleen injury in terms of inflammatory response, oxidative stress, and apoptosis.
Mice ; Animals ; Lipopolysaccharides/pharmacology* ; Spleen ; Inflammation ; Apoptosis ; Sepsis ; Oxidative Stress

ObjectiveTo determine the seroprevalence of anti-Streptococcus pneumoniae immunoglobulin G （IgG） antibody and its risk factors in the elderly attending physical examination. MethodsTwo communities were randomly selected， in which the elderly aged over 60 years attending physical examination in the community health care centers were included in the study. A questionnaire survey was conducted， companied by examination for serum anti-Streptococcus pneumoniae IgG antibody. In addition， data of physical examination was collected. Chi-square test and multivariate logistic regression were utilized to determine the risk factors associated with seroprevalence of anti-Streptococcus pneumoniae antibody. ResultsA total of 635 participants were included. The average age was （73.2±7.4） years old. Forty-nine （7.72%） participants tested positive for anti-Streptococcus pneumoniae IgG antibody. The seroprevalence differed by gender （6.49% in male and 8.86% in female）， smoking （5.48% in smokers and 8.01% in non-smokers）， history of pneumonia （7.89% in those suffered from pneumonia in the past year and 7.71% in those didn^’t have pneumonia）， history of common cold （10.38% in those had common cold in the past year and 5.87% in those didn’t have cold）， obesity （15.62% in those with obesity and 6.83% in those without obesity）. Further multivariate logistic regression showed the seroprevalence of anti-Streptococcus pneumoniae IgG significantly increased in the elderly with history of common cold （OR： 2.24； 95%CI： 1.21‒4.14）and obesity （OR： 2.81； 95%CI： 1.26‒6.26）. ConclusionSeroprevalence of anti-Streptococcus pneumoniae IgG antibody is low in the elderly attending physical examination. It warrants improving the vaccination of pneumococcal polysaccharide vaccine for the elderly with obesity and susceptibility to common cold.

Objective:To compare the clinical efficacy of femoral neck system (FNS) and cannulated screw fixation in the treatment of femoral neck fracture in the young and middle-aged patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 76 young and middle-aged patients with femoral neck fracture admitted to Honghui Hospital Affiliated to Xi′an Jiaotong University from March 2018 to September 2020, including 45 males and 31 females, aged 23-63 years [(49.5±8.2)years]. Garden classification of the fracture was 25 patients with type II, 34 with type III and 17 with type IV. Pauwels classification of the fracture was 16 patients with type I, 31 with type II and 29 with type III. A total of 40 patients were treated with cannulated screw fixation (cannulated screw group) and 36 patients with FNS fixation (FNS group). Operation time, frequency of fluoroscopy, length of hospital stay, weight-bearing time and bone union time were compared between the two groups. Hip function was assessed by Harris hip score at 6, 12 months after operation. The incidence of postoperative complications was observed.Results:All patients were followed up for 12-20 months [(15.2±1.7)months]. The frequency of fluoroscopy [(13.4±1.9)times], weight-bearing time [(11.1±1.9)weeks] and bone union time [(13.8±1.6)weeks] in FNS group were lower or shorter than those in cannulated screw group [(18.2±2.6)times, (15.7±1.6)weeks, (14.6±1.6)weeks] (all P<0.05 or 0.01). There were no significant differences in operation time, length of hospital stay and Harris hip score at 6, 12 months after operation between the two groups (all P>0.05). No postoperative infection occurred in both groups. In cannulated screw group, there were 4 patients with femoral neck shortening, 3 with deep venous thrombosis, 2 with internal fixation loosening, 2 with nail withdrawal and 1 with fracture non-union, showing the incidence of complications of 30% (12/40). In FNS group, there was 2 patients with femoral neck shortening, 1 with deep vein thrombosis and 1 with internal fixation loosening, showing the incidence of complications of 11% (4/36) ( P<0.05). Conclusion:For femoral neck fracture in the young and middle-aged patients, both FNS and cannulated screw fixation can obtain satisfactory efficacy, but FNS has the advantages of decreased frequency of intraoperative fluoroscopy, early weight bearing, reduced time to bone union and low incidence of complications.

Objective:To explore the immune infiltration cells in rheumatoid arthritis (RA) synovial lesions, and to provide new research directions and therapeutic targets for the pathogenesis and treatment of RA.Methods:The three gene expression data sets GSE77298, GSE55457 and GSE1919 were downloaded from gene expression omnibus (GEO) (http://www.ncbi.nlm.nih.gov/geo), and the data were merged with Perl. The "limma" package was used to adjust batch differences. In R, "CIBERSORT" software was used to obtain the expression matrix of 22 kinds of immune cells corresponding to RA synovial tissue samples and normal synovial tissue samples were analyzed with the three packages of "e1071", "parallel" and "preprocessCore". Perl was used to screen samples with P<0.05 in the immune cell matrix. R's "barplot" function was analyzed by the percentage of 22 immune cells in samples with P<0.05. The "pheatmap" package of R was used to visualize heatmaps, and "corrplot" package was used to draw correlation heatmaps. The "vioplot" package of R was used to draw violin plots of differences via the wilcox test. Results:The results of immune cell infiltration analysis showed that in RA synovial tissue samples and normal synovial tissue samples at P<0.05, B cells naive and natural killer cells resting were under-expressed in RA synovial tissue, and plasma cells, mast cells resting, macrophages M1, B cells memory and T cells regulatory were highly expressed in RA synovial tissue. This study also found that in the same sample, the correlation coefficient between natural killer cells resting and neutrophils ( r=0.91) was the highest, indicating synergistic effect between the two. In the same sample, the correlation coefficient between macrophages M0 and plasma cells ( r=-0.88) was the lowest, indicating antagonistic effect between the two. Conclusion:The immune infiltrating cells in RA synovial lesions discovered in this study provide a certain theoretical basis and research direction for the research on the disease mechanism and treatment of RA.