The nucleus pulposus (NP) plays a vital role in intervertebral disc degeneration (IVDD). Previous studies have revealed cellular heterogeneity in NP tissue during IVDD progression. However, the cellular and molecular alterations of diverse cell clusters during IVDD remain to be fully elucidated. NP tissues were isolated from patients with different grades of IVDD undergoing discectomy, and then subjected to single-cell RNA sequencing (scRNA-seq). Cell subsets were identified based on unbiased clustering of gene expression profiles. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to determine the molecular features of diverse cell clusters. Monocle analysis was used to illustrate the differentiation trajectories of chondrocytes. Additionally, CellPhoneDB analysis revealed potential interactions between chondrocytes and other cells during IVDD. Based on the expression profiles of 47,610 individual cells, eight putative clusters including chondrocytes, endothelial cells, fibroblasts, macrophages, mural cells, osteoclasts, proliferating stromal cells and T cells were identified. The chondrocyte cluster was classified into three subsets, C1-C3, which were associated with stress-resistance, fibrosis and inflammatory responses, respectively. Pseudo-time trajectories suggested that chondrocytes gradually differentiated into fibroblasts during IVDD. Immune cells including cDC2s, macrophages and monocytes were identified. Further analysis showed that chondrocytes might communicate with immune cells via the MIF, TNFSF9, SPP1 and CCL4L2 signaling pathways. In addition, we found that invading endothelial cells might interact with chondrocytes through the COL4A1, CXCL12, VEGFA and SEMA3E signaling pathways. Our results reveal the cellular complexity and phenotypic characteristics of NP tissues at single-cell resolution, which will contribute to the in-depth investigation of preventative and regenerative strategies for IVDD.

Intervertebral disc degeneration (IDD) leads to low back pain (LBP). This study aimed to determine the regulation of IDD by competing endogenous RNAs (ceRNAs). We obtained the GSE63492, GSE124272, and GSE129789 datasets from the Gene Expression Omnibus database. The changes of long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs in IDD were characterized. The significantly changed mRNAs were subjected to protein-protein interaction analysis using the STRING database, and its functions and involved pathways were analyzed using the DAVID database and gene set enrichment analysis (GSEA). The significant changed lncRNAs, miRNAs and mRNAs were linked in a ceRNA network based on their interactions - predicted by Starbase and miRWalk. Differentially methylated loci of significantly changed mRNAs in early and advanced IDD were compared using the GSE129789 dataset. We identified 245 significantly changed mRNAs, 133 lncRNAs, and 228 miRNAs between patients with IDD and normal individuals. GSEA suggested that 17 pathways related to cell proliferation were activated while 35 cell signaling and immune-related pathways were suppressed in IDD. The following ceRNA network in IDD was built: LINC00665/hsa-miR-7-5p/FZD3, ZNF549; LINC00665/hsa-let-7e-5p/FZD3, ACVR2B; TRG-AS1/hsa-miR-574-5p/ACVR2B, P3H2; TRG-AS1/ hsa-let-7e-5p/FZD3, ACVR2B; and ZNF571-AS1/let-7e-5p/ACVR2B, FZD3. A lncRNA-miRNAmRNA ceRNA network which might regulate the progression of IDD was developed.

[目的]通过生物力学研究,分析3种类型股骨粗隆间骨折内侧阳性、中性及阴性支撑下的稳定性差异.[方法]使用SYNBONE人工股骨标准试验骨分别制作A1.1/A2.1/A3.2 3种粗隆间骨折模型,并使用股骨近端抗旋髓内固定,根据不同的复位状态分为阳性、中性及阴性支撑组,通过静态轴向压缩试验分析在不同载荷下的股骨近端下沉位移值以及模型轴向刚度值.[结果]A1.1型骨折阳性及中性支撑模式股骨近端下沉位移均小于阴性支撑,且前2模式的轴向刚度均大于阴性支撑模式(P＜0.05).A2.1型骨折股骨近端下沉位移依次为:阳性支撑＜中性支撑＜阴性支撑;刚度依次为阳性支撑＞中性支撑＞阴性支撑;组间差异均有统计学意义(P＜0.05).A3.2型骨折中性支撑组位移显著小于阳性支撑及阴性支撑,而刚度显著大于后两者(P＜0.05).[结论]内侧皮质阳性支撑可视为A1.1和A2.1型粗隆间骨折的最优复位状态,而A3.2型反粗隆间骨折应最大程度获得解剖复位中性支撑.

Objective:To analyze the relative expression of guanosine-5′-triphosphate cyclohydrolase 1 (GCH1) in the regulation of circular RNA in mouse microglia (BV2) in vitro. Methods:BV2 cells were cultured and passaged. The BV2 cells were transfected with viruses and divided into a low expression GCH1 adenovirus vector group (Ad-shGCH1, n=3) and a control adenovirus vector group (Ad-NC, n=3). Two sets of samples. Incubate for 48 h to observe the transformation efficiency and extract total RNA with TRIzol reagent. The second-generation circular RNA (circRNA) high-throughput sequencing was used to analyze the Ad-shGCH1 and Ad-NC samples to screen for differentially expressed circular circRNA. CircRNA sequencing data uses the R package edge R to calculate the fisher exact test based on the negative binomial distribution model to determine the significance of the difference. Finally, the coding-non-coding gene co-expression (CNC) network analysis was used to conduct bioinformatics analysis on circRNAs with significant expression differences, and finally verified by reverse transcription-polymerase chain reaction(RT-PCR). Results:Compared with the control group, we found that mmu_circ_0001322 ( P<0.05), mmu_circ_0000454 ( P<0.05), mmu_circ_0001383 ( P<0.05), mmu_circ_0000898( P<0.05) were up-regulated and mmu_circ_0000652 was down-regulated (Multiple change> 1.2 times) in Ad-shGCH1. The CNC analyzed that mmu_circ_0000652 may play a role by participating in the regulation of mitogen-activated protein kinase14(MAPK14) by participating in specific network or biological processes. Conclusion:CircRNA regulated by GCH1 may be involved in the inflammatory process of microglia in vitro, and may provide new information for the treatment of neuropathic pain.

Brucella abortus is highly contagious and can result in abortion or infertility in animals. This study investigated the mechanism of B. abortus-induced mucosal immunity at different points after infection in a mouse model. GSE125765 was obtained from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) in nasal-associated lymphoid tissues at 1, 6, and 12 h after intranasal immunization, compared with control samples, were screened. Functional enrichment analysis was performed using the DAVID tool. Gene modules associated with the sample phenotype were then explored using WGCNA. Mfuzz was used to analyze the expression pattern trends of DEGs. The KEGG pathway directly related to B. abortus infection was searched based on the Comparative Toxicogenomics Database 2019 update. Three gene modules, 172, 366, and 167 DEGs aggregated into four expression patterns, were significantly positively correlated with B. abortus infection. The expression of DEGs in cluster 1 first increased and then decreased with prolonged infection. DEG expression in clusters 2-4 continued to rise with prolonged infection. Genes related to B. abortus infection were significantly enriched in 'lysosome' in the Comparative Toxicogenomics Database, including genes CTSL, CD68, SCARB2, and CTSS, which might be necessary for modulating host-B. abortus interactions through the 'lysosome' pathway.

[目的]分析脂多糖诱导小胶质细胞中炎症相关的长链非编码RNA (long non-coding RNA,lncRNA)表达.[方法]将小鼠小胶质细胞(BY2)样本随机分为两组,脂多糖(lipopolysaccharide,LP)1ug/ml处理24h后细胞组为LPS组,未做LPS干预细胞为对照组.通过生物信息学分析后,实时荧光定量PCR (Quantitative Real-time PCR,qPCR)进一步验证LPS诱导后BV2的lncRNA与炎症的关系.[结果]在高通量测序中选取炎症相关差异表达明显的9个lncRNA,qPCR验证后发现AC005899.1、GPATCH8、VPS41与高通量测序中的表达趋势一致.[结论]多个lncRNA可能参与体外小胶质细胞的炎症反应过程.这些结果为小胶质细胞通过lncRNA参与神经病理性疼痛提供相关信息.

Several studies have confirmed that microglia are involved in neuropathic pain.Inhibition of guanosine-5′-triphosphate cyclohydrolase 1(GTPCH1) can reduce the inflammation of microglia.However,the precise mechanism by which GTPCH1 regulates neuropathic pain remains unclear.In this study,BV2 microglia were transfected with adenovirus to knockdown GTPCH1 expression.High throughput sequencing analysis revealed that the mitogen-activated protein kinase(MAPK) related pathways and proteins were the most significantly downregulated molecular function.Co-expression network analysis of Mapk14 mRNA and five long noncoding RNAs(lnc RNAs) revealed their correlation.Quantitative reverse transcription-polymerase chain reaction revealed that among five lnc RNAs,ENSMUST00000205634,ENSMUST00000218450 and ENSMUST00000156079 were related to the downregulation of Mapk14 mRNA expression.These provide some new potential targets for the involvement of GTPCH1 in neuropathic pain.This study is the first to note the differential expression of lnc RNAs and mRNA in GTPCH1 knockdown BV2 microglia.Findings from this study reveal the mechanism by which GTPCH1 activates microglia and provide new potential targets for microglial activation in neuropathic pain.

神经性疼痛是由多种疾病或神经系统损伤引起的慢性疼痛,因其发病机制尚不明确,神经性疼痛往往比非神经性疼痛更难治愈.目前在病理性疼痛发展和维持的过程中,外周致敏和中枢致敏可被看作神经元可塑性的表现.在这两个方面中,细胞因子和趋化因子在介导疼痛的发生发展中起到关键的作用.最近的研究表明炎性趋化因子CXCL1在脊髓胶质细胞中的表达与神经性疼痛密切相关. CXCL1是一种促进中枢致敏作用的趋化因子,其与受体CXCR2结合后介导神经性疼痛.现对CXCL1/CXCR2与神经性疼痛的关系作一综述.

Lumbar disc herniation (LDH) is one of the most common degenerative spinal diseases in adults and less common in adolescents.The age of onset is less than 21 years old and is defined as adolescent lumbar disc herniation (ALDH).The incidence of ALDH is only 1％-5％.However,with the accelerated pace of life,the incidence of ALDH is increasing.In recent years,the rapid development of endoscopic technology has been widely used in spinal surgery.ALDH's percutaneous endoscopic technique has achieved good results.Compared with open surgery,percutaneous endoscopic treatment can preserve more normal lumbar structures,such as ligaments,muscles,lamina and facets,minimizing the impact on pubertal growth.This article reviews the characteristics of ALDH and endoscopic treatment as an entry point.

Neuropathic pain is caused by somatosensory system injury or disease.The prevalence of neuropathic pain in the general population is about 6.9％-10.0％ which produces bad effects to quality of life of patients and imposes great burden on individuals,families and society.Many factors could lead to neuropathic pain,such as injuries and congenital diseases.Therefore,exploring the molecular targets of neuropathic pain treatment is of great significance.In recent years,scholars have explored new ways about targeted therapy of neuropathic pain by regulating long non-coding RNA (lncRNA) and studying the composition of traditional Chinese medicine.The janus kinase (JAK)/signal transducer and activators of transcription 3 (STAT3) pathway is one of the important components that involved in neuropathic pain,but specific mechanism is still unclear.This article reviews the recent advances in JAK/ STAT3 and neuropathic pain.

神经性疼痛是一种难治性慢性疾病,严重危害患者的生命质量,并且对社会及家庭带来沉重负担,我国乃至全球每年治疗慢性疼痛的花费数以亿计.然而目前常规镇痛药物难以达到理想的止痛效果,因此亟需开发新靶点治疗神经性疼痛.长链非编码RNA(lncRNA)正在成为基因调控的新参与者,但是lncRNA如何参与神经病理性疼痛的机制尚不清楚.虽然lncRNA最初被认为是转录副产物,但随着最近技术的进步使我们重新认识它们在基因调控和疾病发病机制中的重要作用.目前研究的热点从最初分析miRNA的表达与调控逐步转向lncRNA及其介导的疼痛调节功能机制,本文以此做以综述深度剖析lncRNA与神经性疼痛的关系.