The auxin/indole-3-acetic acid (Aux/IAA) gene family is an important regulator for plant growth hormone signaling, involved in plant growth, development, as well as response to environmental stresses. In the present study, we identified SmIAA7 which is potentially associated with Salvia miltiorrhiza leaf development through comparatively analyzed the transcriptome data from different pinnate leaves. SmIAA7 was successfully isolated from S. miltiorrhiza using the specific primers. Then subsequent bioinformatic analysis, prokaryotic expression and purification, subcellular localization, and induction expression analysis under auxin and abiotic stress were performed. The full-length of SmIAA7 contained an ORF of 684 bp encoding a protein of 227 amino acid with a molecular weight of 25.3 kD. Conserved domain analysis showed that SmIAA7 contains the conserved Aux_IAA domain (pfam02309). Sequence analysis and phylogenetic tree analysis results indicated SmIAA7 was phylogenetically close to IAA7 and IAA14 from other plants, suggesting SmIAA7 involved in plant growth and development as well as response to environmental stresses. The prokaryotic expression vector pET28a-SmIAA7 was constructed and SmIAA7 recombinant protein was successfully expressed in E. coli Rosetta (DE3) strain. Subcellular localization experiment demonstrated that SmIAA7 localized in the nucleus of plant cells. Real-time fluorescence quantitative PCR results showed that the expression level of SmIAA7 was upregulated in response to auxin. Drought, low temperature, and salt stress significantly increased the transcript level of SmIAA7 gene. This study lays a foundation for further elucidating the role of SmIAA7 in leaf development, signal transduction, and stress defense in S. miltiorrhiza.

ObjectiveTo study the regulatory effect of the partial sequence within the 3’ untranslated region （3’UTR） of slit guidance ligand 1 （Slit1） （Slit1-3’UTR） on the fibrotic phenotypes of cardiac fibroblasts （CFs） and its potential mechanism. MethodsThe adenovirus vector was used to overexpress the 1526nt sequence of Slit1-3’UTR in ICR neonatal mouse CFs （mCFs）. The expression of fibrosis-related genes in mCFs， such as collagen type 1 alpha1（COL1A1）， collagen type 3 alpha3 （COL3A1） and alpha smooth muscle actin （α-SMA） were detected by Western blot assay. The effect of Slit1-3’UTR 1526nt on the proliferation and migration of mCFs was assessed by EdU staining and Trans-well assays. Angiotensin Ⅱ （Ang Ⅱ） was used to treat mCFs， and the impact of Slit1-3’UTR 1526nt on the fibrotic phenotypes of Ang Ⅱ-induced mCFs was evaluated. After overexpression of Slit1-3’UTR 1526nt， miR-34a-5p mimic was transfected into mCFs， followed by actinomycin D treatment to detect the mRNA stability of Slit1-3’UTR 1526nt， and the levels of miR-34a-5p and its target gene SIRT1（si-SIRT1） in mCFs were determined. The effects of miR-34a-5p and small interfering RNA targeting SIRT1 on the Slit1-3’UTR 1526nt-mediated regulation of fibrotic phenotypes were also determined. ResultsAdenovirus-mediated overexpression of Slit 1-3’UTR 1526nt was achieved in mCFs. Overexpression of Slit 1-3’UTR 1526nt markedly inhibited the expression of the fibrosis-related genes， proliferation and migration of mCFs and fibrotic phenotypes of Ang Ⅱ. The results of actinomycin D assay showed that miR-34a-5p inhibited the stability of Slit1-3’UTR 1526nt in mCFs， while the level of miR-34a-5p was reduced in mCFs with overexpression of Slit1-3’UTR 1526nt. Transfection of miR-34a-5p promoted the fibrotic phenotypes， and reversed the inhibitory effect of Slit1-3’UTR 1526nt on the fibrotic phenotypes of mCFs. Overexpression of Slit1-3’UTR 1526nt significantly increased the level of miR-34a-5p target gene SIRT1 in mCFs. Transfection of miR-34a-5p and si-SIRT1 consistently reversed the inhibitory effects of Slit1-3’UTR 1526nt on the fibrotic phenotypes of mCFs. ConclusionSlit1-3’UTR1526nt inhibits the fibrotic phenotypes of mCFs by binding to miR-34a-5p and increasing the expression of its target gene of SIRT1.

BACKGROUND:Increasing evidence suggests that N6-methyladenosine(m6A)regulators are closely associated with osteoarthritis and are considered to be a new direction in the prevention and treatment of osteoarthritis,but their specific mechanism of action is unknown. OBJECTIVE:To conduct a bioinformatics analysis of the osteoarthritis gene microarray dataset in order to explore the role of m6A in osteoarthritis and analyze the pathogenesis of osteoarthritis. METHODS:The m6A regulators associated with osteoarthritis and their expression were first extracted from the GSE1919 dataset in the GEO database using R software,and then the results were analyzed by gene difference analysis and GO and KEGG enrichment analyses.Subsequently,the results of protein-protein interaction network topology analysis and machine learning results were intersected to obtain the m6A Hub regulators,which were validated by in vitro cellular experiments. RESULTS AND CONCLUSION:A total of 16 osteoarthritis-related m6A regulators were extracted and 11 m6A differential regulators,including ZC3H13,YTHDC1,YTHDF3 and HNRNPC,were obtained by differential analysis.GO enrichment analysis showed that osteoarthritis-related m6A differential regulators played a role in the biological processes such as mRNA transport,RNA catabolism,and regulation of insulin-like growth factor receptor signaling pathway.(3)KEGG enrichment analysis showed that the differential regulators were mainly involved in the p53,interleukin-17 and AMPK signaling pathways.The combined protein-protein interaction network topology analysis and machine learning results obtained the m6A Hub regulator-YTHDC1.(5)The results of in vitro cellular experiments showed that there was a significant difference in the expression of m6A key regulator between the control and experimental groups(P<0.05).To conclude,YTHDC1 is closely related to the development of osteoarthritis,which is expected to be a molecular target of m6A for the treatment of osteoarthritis.

BACKGROUND:SMAD family member 4(SMAD4)can promote bone remodeling in osteoporotic rats,but it is unclear whether SMAD4 interferes with the expression of iron metabolism related proteins in osteoporotic rats. OBJECTIVE:To explore the effect of SMAD4 overexpression on the expression of iron metabolism related proteins in osteoporotic rats. METHODS:Rats were randomized into sham group,ovariectomy group,transfection control group and SMAD4 overexpression group.Animal models of osteoporosis were established in the latter three groups by ovariectomy,and only adipose tissue was removed in the sham group.One week later,adenovirus was injected into the femoral bone marrow cavity.SMAD4 overexpression group and transfection control group were injected with adenovirus overexpressing SMAD4 gene and control empty virus,respectively.Index detection was performed at 1 month after injection.Micro-CT,hematoxylin-eosin staining and tartrate resistant acid phosphatase staining were used to detect bone formation and bone resorption in osteoporotic rats.ELISA was used to detect serum ferritin and hepcidin levels.Immunohistochemical staining was used to detect alkaline phosphatase,osteocalcin,receptor activator for nuclear factor-κB ligand and tartrate resistant acid phosphatase levels in femoral tissue.RT-qPCR was used to detect SMAD4,hepcidin,divalent metal transporter 1,transferrin receptor1 and ferroportin1 mRNA levels in femoral tissue.Western blot was used to detect SMAD4,alkaline phosphatase,osteocalcin,osteoprotegerin,receptor activator for nuclear factor-κB ligand,tartrate resistant acid phosphatase,β-Crosslaps,hepcidin,divalent metal transporter 1,transferrin receptor 1,and ferroportin 1 protein levels. RESULTS AND CONCLUSION:In the sham group,bone trabeculae in femur tissue were intact,and almost no osteoclasts were found.In the ovariectomy and transfection control groups,the bone trabeculae were sparse and a large number of osteoclasts were present.In the SMAD4 overexpression group,the number of bone trabeculae was increased and the number of osteoclasts was decreased.Compared with the sham group,the ovariectomy group showed a significant reduction in the protein expression of SMAD4,alkaline phosphatase,osteocalcin,and osteoprotegerin in femoral tissue and hepcidin levels in serum and femoral tissue,while receptor activator for nuclear factor-κB ligand,tartrate resistant acid phosphatase,β-Crosslaps protein levels,divalent metal transporter 1,transferrin receptor1,ferroportin1 mRNA and protein levels were significantly increased(P＜0.05).Compared with the transfection control group,the SMAD4 overexpression showed a significant increase in SMAD4,alkaline phosphatase,osteocalcin,and osteoprotegerin protein levels in femoral tissue and hepcidin levels in serum and femoral tissue,while the expressions of receptor activator for nuclear factor-κB ligand,tartrate resistant acid phosphatase,β-Crosslaps protein levels,divalent metal transporter 1,transferrin receptor1,and ferroportin 1 at mRNA and protein levels were significantly decreased(P＜0.05).To conclude,overexpression of SMAD4 promotes bone remodeling in osteoporotic rats by interfering with the expression of iron metabolism related proteins.

BACKGROUND:How to improve the accuracy of puncture,reduce surgical damage,and improve surgical efficiency during vertebroplasty is currently one of the focuses of exploration and improvement in vertebroplasty techniques. OBJECTIVE:To explore the clinical significance of application of mixed reality technology in percutaneous vertebroplasty for spinal fractures. METHODS:Two patients with osteoporotic vertebral compression fracture in Dalian Second People's Hospital in June 2023 were selected.Before operation,128-row CT scanning of the lumbar spine was performed and the original data of digital imaging and communications in medicine(DICOM)were obtained.Visual Volume software was used to build the three-dimensional network model of vertebral compression fracture.Holographic imaging glasses were used to accurately map 3D network model images to the real world,assist the surgeon in completing preoperative simulation,explaining preoperative conditions and treatment plans,and guiding puncture and bone cement injection during surgery. RESULTS AND CONCLUSION:(1)Precise puncture was achieved with the assistance of a mixed reality technology.Postoperative imaging examination showed good bone cement filling and no obvious leakage.The postoperative symptoms of the patient were alleviated well,and they were able to move to the ground on the same day after surgery.(2)It is concluded that a mixed reality technology is helpful for preoperative surgical design and communication efficiency with patients and their families.Assisting with precise puncture during surgery,shortening surgical time,and reducing side injuries is a new and effective clinical diagnosis and treatment model,which has development potential in minimally invasive,precise,and personalized treatment of spinal surgery.

BACKGROUND:Scoliosis mainly refers to sequence abnormalities in the coronal,sagittal,and axial positions of the spine,with a Cobb angle of≥10°.The patients may experience symptoms such as unequal shoulder height and back asymmetry.Severe cases may affect the patient's cardiopulmonary function,thereby affecting their daily life.Conservative treatment can control the progression of scoliosis and avoid later surgery.Scoliosis orthosis is currently a commonly used and effective treatment measure in conservative treatment. OBJECTIVE:To summarize and analyze the current research status,hotspots,and trends of scoliosis orthoses both domestically and internationally,providing reference for related research. METHODS:Using bibliometrics and visual analysis as tools,and using a comparison between China and foreign countries as a method,this paper analyzes the literature on scoliosis orthosis journals in the past decade.Based on bibliometrics,the current status of research on scoliosis orthoses is determined.Citespace software is used to analyze key words and identify the current hotspots and future trends in scoliosis orthosis research. RESULTS AND CONCLUSION:(1)At present,the number of literature on scoliosis orthoses is still on a fluctuating upward trend.China and the United States are the main countries for research,with a literature share of over 40%.However,the average citation rate of foreign language literature by Chinese scholars is relatively low.(2)The basic fields of domestic research are mainly surgery and pediatrics,while orthotics and clinical neurology are mainly studied abroad.Among them,there is also a certain number of documents in domestic Chinese medicine,indicating that China is also engaged in the combination of Chinese and Western treatment of scoliosis.The National Natural Science Foundation of China has the highest proportion in the aspect of Chinese and foreign literature,reflecting the importance of the fund attaches to the research of scoliosis orthosis.(3)The authors with the highest number of publications are Qiu Yong and Negrini Stefano,and the most published institutions are the Spinal Surgery Department of Gulou Hospital affiliated to Nanjing University Medical College and UDICE-French Research University.Domestic and foreign authors and institutions have certain communications about this,but not closely,which requires relevant institutions and scholars to further explore and study.(4)From the research hotspots and future trends,the main treatment type is adolescent idiopathic scoliosis,while the production method of the short-column side bending orthosis is three-dimensinoal printing,and the main treatment index is convex progression.The ultimate purpose of treatment is to improve the quality of life of the patients.

Cellulose synthase (CesA), one of the key enzymes in the biosynthesis of cellulose in plants, plays an important role in plant growth and plant resistance. In this study, a total of 21 AsCesA genes from Aquilaria sinensis were systematically identified and the physico-chemical characteristics were analyzed based on genome database and bioinformatical methods. The phylogenetic tree was constructed and the gene location on chromosome, cis-acting elements in the 2 000 basepairs upstream regulatory regions and conservative motifs were analyzed. The AsCesA proteins were mainly located on the plasma membrane. The number of amino acids of the proteins ranged from 390 to 1 261. The isoelectric point distributed from 5.67 to 8.86. All of the 21 AsCesA proteins possessed the transmembrane domains, the number of which was from 6 to 8. The genes were classified into 3 groups according to the phylogenetic relationship. Obvious differences were observed in motif composition in genes from different groups. However, motif2, motif6, motif7 and motif10 were observed in all of AsCesA proteins. Analysis of cis-acting elements indicated that AsCesA genes family has cis-acting elements related to plant hormones, abiotic stresses, and biological processes. Seven AsCesA genes with differential expression were selected according to the calli transcriptome data induced by NaCl at different times and their expression levels under different abiotic stresses were analyzed by quantitative real-time PCR. The results indicated that salt, low temperature, drought, and heavy metal stresses could affect the expression level of AsCesA genes, and the abundance of AsCesA1, AsCesA3 and AsCesA20 showed a significant change, implying their potential important roles to the abiotic stresses. The accumulation pattern of cellulose content under different abiotic stresses was similar to the expression trend of AsCesA genes. Our results provide valuable insights into the role of cellulose synthase in A.sinensis in plant defense.

Platycodonis Radix is the dry root of Platycodon grandiflorum of Campanulaceae, which has a variety of pharmacological effects and is a commonly used bulk Chinese medicine. In this study, the chloroplast genome sequences of six P. grandiflorum from different producing areas has been sequenced with Illumina HiSeq X Ten platform. The specific DNA barcodes were screened, and the germplasm resources and genetic diversity were analyzed according to the specific barcodes. The total length of the chloroplast genome of 6 P. grandiflorum samples was 172 260-172 275 bp, and all chloroplast genomes showed a typical circular tetrad structure and encoded 141 genes. The comparative genomics analysis and results of amplification efficiency demonstrated that trnG-UCC and ndhG_ndhF were the potential specific DNA barcodes for identification the germplasm resources of P. grandiflorum. A total of 305 P. grandiflorum samples were collected from 15 production areas in 9 provinces, for which the fragments of trnG-UCC and ndhG_ndhF were amplificated and the sequences were analyzed. The results showed that trnG-UCC and ndhG_ndhF have 5 and 11 mutation sites, respectively, and 5 and 7 haplotypes were identified, respectively. The combined analysis of the two sequences formed 13 haplotypes (named Hap1-Hap13), and Hap4 is the main genotype, followed by Hap1. The unique haplotypes possessed by the three producing areas can be used as DNA molecular tags in this area to distinguish from the germplasm resources of P. grandiflorum from other areas. The haplotype diversity, nucleotide diversity and genetic distance were 0.94, 4.79×10^-3 and 0.000 0-0.020 3, respectively, suggesting that the genetic diversity was abundant and intraspecific kinship was relatively close. This study laid a foundation for the identification of P. grandiflorum, the protection and utilization of germplasm resources, and molecular breeding.

Objective     To investigate the relationship between preoperative mean daily step counts and pulmonary complications after thoracoscopic lobectomy in elderly patients. Methods     From 2018 to 2021, the elderly patients with pulmonary complications after thoracoscopic lobectomy were included. A 1∶1 propensity score matching was performed with patients without pulmonary complications. The clinical data were compared between the two groups. Results    Totally, 100 elderly patients with pulmonary complications were enrolled, including 78 males and 22 females, aged 66.4±4.5 years. And 100 patients without pulmonary complications were matched, including 71 males and 29 females aged 66.2±5.0 years. There was no significant difference in the preoperative data between the two groups (P>0.05). Compared to the patients with pulmonary complications, the ICU stay was shorter (8.1±4.4 h vs. 12.9±7.5 h, P<0.001), the first out-of-bed activity time was earlier (8.8±4.5 h vs. 11.2±6.1 h, P=0.002), and the tube incubation time was shorter (19.3±9.2 h vs. 22.5±9.4 h, P=0.015) in the patients wihout pulmonary complications. There was no statistical difference in other perioperative data between the two groups (P>0.05). The mean daily step counts in the pulmonary complications group were significantly less than that in the non-pulmonary complications group (4 745.5±2 190.9 steps vs. 6 821.1± 2 542.0 steps, P<0.001). The daily step counts showed an upward trend for three consecutive days in the two groups, but the difference was not significant. Conclusion     The decline of preoperative mean daily step counts is related to pulmonary complications after thoracoscopic lobectomy in elderly patients. Recording daily step counts can promote preoperative active exercise training for hospitalized patients.

Objective To investigate the effect of AU-rich element RNA-binding factor 1(AUF1)on the proliferation,apoptosis,and migration abilities of hepatocellular carcinoma(HCC)cells and possible mechanisms,and to clarify the role and molecular mechanism of AUF1 in the progression of HCC.Methods The UALCAN and TCGA-HCC databases were used to analyze the expression of AUF1 in pan-cancer and investigate the association of the expression level of AUF1 with the clinicopathological features and prognosis of HCC patients.CCK-8 assay,cell apoptosis assay,and Transwell chamber assay were used to investigate the function of AUF1 at the cellular level,and RNA-seq assay was used to investigate transcriptome changes in HCC cells after AUF1 knockdown.The t-test was used for comparison of continuous data between two groups;the Kaplan-Meier method was used to plot survival curves,and the log-rank test was used for comparison of survival rates.Results There were abnormal mRNA and protein expression levels of AUF1 in various tumor tissues compared with normal tissue(P<0.05).The mRNA expression level of AUF1 was positively correlated with the degree of HCC malignancy and the poor prognosis of early-stage HCC(P<0.05).Compared with the control group,the overexpression of exogenous AUF1 in HCC cells promoted the proliferation of HCC cells and inhibited the apoptosis and migration of HCC cells,while AUF1 knockdown inhibited HCC cell proliferation and promoted the apoptosis and migration of HCC cells.The RNA-seq analysis showed that AUF1 knockdown mainly affected the Wnt/β-catenin pathway and downregulated the protein expression level of β-catenin.Conclusion The abnormal expression of AUF1 is associated with the prognosis of early-stage HCC,and AUF1 may exert an oncogenic effect by activating the Wnt signaling pathway.