Objective To investigate the biocompatilility of human acellular amniotic membrane(HAM) with vascular smooth muscle cells(VSMCs) and to explore the possibility to construct tissue engineering bladder with HAM as the scaffold and VSMCs as the seed cells. Methods After physical and 1% trypsogen preparation,the HAM was mixed with VSMCs taken from rats for culture in vitro.Histological obserbation was done under inverted microscope and scanning electron microscope respectively.20 rats were divided into two groups.Hemicystectomies were performed in 20 rats,and 10 of them were repaired with HAM grafts with VSMCs on the half bladder,the other 10 were repaired with HAM grafts without VSMCs as the control group.The rats underwent postoperative assessment of bladder volume at the 2nd,4th and 8th weeks,and the grafts were observed by light microscope at the 2nd,4th and 8th weeks after surgery. Results The physical and 1% trypsogen treated HAM was pure with hollows and undamaged collagen fibers.The VSMCs could grow,adhere to and differentiate on the surface of HAM and into the hollows.At the 2nd,4th and 8th weeks after surgery, the bladder volumes of the experimental group were not different significantly compared with those of the control group.Epithelialization and smooth muscle cells regeneration occurred with the infiltration of inflammatory cells in the 2nd week after grafting,and the HAM were absorbed.In the 4th and 8th weeks,it was difficult to delineate the junction between the host bladder and grafts by histology.Conclusion HAM can be used as the scaffold to construct tissue engineering bladder as it has good biocompatibility with VSMCs without disturbing the cell form and the graft can be absorbed quickly.

Objective: To evaluate the effects of orthopedic robot navigation system used in core decompression combined with bone graft surgery for osteonecrosis of the femoral head (ONFH). Methods: A retrospective analysis was performed on 9 patients (16 hips) underwent core decompression surgery combined with bone graft treatment for early ONFH in ARCO Ⅱat Department of Osteoarthropathy, Yantaishan Hospital from June 2016 to February 2018.There were 7 males and 2 females, aged 44.6 years (range: 28-60 years). All surgery procedures were completed by the same doctor under the navigation of orthopedic surgery robot using the specific designed tools. The preoperative and postoperative Harris Hip Score and visual analogue score were compared and the time of intraoperative X-ray fluoroscopy was recorded. Results: The mean follow-up period was (12.7±3.5)months (range: 6-18 months). Four cases of treatment failure were observed in 16 hips at the last follow-up and the hip survival rate was 12/16.In stage ARCO Ⅱa, Ⅱb and Ⅱc, the survival rate was 1/1, 2/2, and 9/13.The Harris Hip Score (88.3±1.9 vs.70.0±3.8, t=16.81, P=0.000) and visual analogue score (3.7±0.7 vs. 0.9±0.6, t=13.49, P=0.000) were improved significantly at the last follow-up compared with preoperative value. The total times of intraoperative X-ray fluoroscopy were 15.9±2.5, including 5.8±1.2 before the insertion of the guiding wire and 10.1±1.7 after the insertion of the guiding wire. Conclusions The domestic orthopedic surgery robot can be applied successfully in the core decompression combined with bone graft surgery for ONFH . It is less invasive with less fluoroscopy time and better bone graft effects.

Ankle osteoarthritis (OA),a progressive disease,will develop into its full end-stage in the long run if no appropriate treatment is given.Joint-sacrificing procedures,such as total ankle replacement and ankle arthrodesis,used to be mostly applied for treatment of painful mid-and end-stage ankle OA,leading to adverse effects in functional recovery,complications and treatment costs.In recent years,joint-preserving surgery has emerged as an increasingly common treatment for mid-stage ankle OA at home and abroad.It has become a hot topic in the field of foot and ankle surgery,because it may restore normal biomechanics of the ankle joint,substantially relieve postoperative pain,improve function,and prevent or slow the degeneration process.This paper reviews the new progress in joint-preserving treatment of ankle OA.

Objective:To explore the relationship between the expression of SF3B1, UBE2V2, SETD2 and osteoporotic vertebral fracture (OVF) in elderly patients.Methods:Peripheral blood samples were collected from 31 elderly patients with osteoporotic vertebral fractures (VF group) and 16 elderly patients with osteoporotic non-vertebral fractures (NVF group) in Yantai Mountain Hospital. RNA was extracted for transcriptome sequencing to screen for differentially expressed genes. VF related genes were screened by Gene Ontology (GO) analysis, protein protein interaction (PPI) network analysis and ROC curve analysis. Qrt-pcr was used to detect gene expression levels.Results:Compared with NVF group, 691 genes were up-regulated while 131 genes were down regulatedin VF group. qRT-PCR results revealed that, compared with NVF patients (1.55±0.33) (1.70±0.33) (1.64±0.33) , SF3B1 (1.83±0.23) ( t=2.84, P=0.008) , UBE2V2 (2.24±0.43) ( t=3.91, P<0.001) expression were increased while SETD2 (1.18±0.46) ( t=3.25, P=0.003) expression was decreased in peripheral blood of VF patients. ROC curve analysis showed that the AUCs of SF3B1, UBE2V2 and SETD2 in VF were 0.8034 ( P=0.007) , 0.8145 ( P=0.005) and 0.7863 ( P=0.0014) , respectively. Conclusion:SF3B1, UBE2V2 and SETD2 are highly correlated with OVF in elderly patients, and are of great value in the diagnosis and prediction of OVF.

Tunicamycin, a potent reversible translocase I inhibitor, is produced by several Actinomycetes species. The tunicamycin structure is highly unusual, and contains an 11-carbon dialdose sugar and an α, β-1″,11'-glycosidic linkage. Here we report the identification of a gene cluster essential for tunicamycin biosynthesis by high-throughput heterologous expression (HHE) strategy combined with a bioassay. Introduction of the genes into heterologous non-producing Streptomyces hosts results in production of tunicamycin by these strains, demonstrating the role of the genes for the biosynthesis of tunicamycins. Gene disruption experiments coupled with bioinformatic analysis revealed that the tunicamycin gene cluster is minimally composed of 12 genes (tunA-tunL). Amongst these is a putative radical SAM enzyme (Tun B) with a potentially unique role in biosynthetic carbon-carbon bond formation. Hence, a seven-step novel pathway is proposed for tunicamycin biosynthesis. Moreover, two gene clusters for the potential biosynthesis of tunicamycin-like antibiotics were also identified in Streptomyces clavuligerus ATCC 27064 and Actinosynnema mirums DSM 43827. These data provide clarification of the novel mechanisms for tunicamycin biosynthesis, and for the generation of new-designer tunicamycin analogs with selective/enhanced bioactivity via combinatorial biosynthesis strategies.
Actinobacteria ; enzymology ; genetics ; Base Sequence ; Biological Assay ; Carbohydrate Sequence ; Carbohydrates ; biosynthesis ; genetics ; Cloning, Molecular ; Gene Deletion ; Gene Library ; High-Throughput Screening Assays ; Molecular Sequence Data ; Multigene Family ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; Streptomyces ; enzymology ; genetics ; Tunicamycin ; biosynthesis ; chemistry ; genetics

Age-associated changes in immune cells have been linked to an increased risk for infection. However, a global and detailed characterization of the changes that human circulating immune cells undergo with age is lacking. Here, we combined scRNA-seq, mass cytometry and scATAC-seq to compare immune cell types in peripheral blood collected from young and old subjects and patients with COVID-19. We found that the immune cell landscape was reprogrammed with age and was characterized by T cell polarization from naive and memory cells to effector, cytotoxic, exhausted and regulatory cells, along with increased late natural killer cells, age-associated B cells, inflammatory monocytes and age-associated dendritic cells. In addition, the expression of genes, which were implicated in coronavirus susceptibility, was upregulated in a cell subtype-specific manner with age. Notably, COVID-19 promoted age-induced immune cell polarization and gene expression related to inflammation and cellular senescence. Therefore, these findings suggest that a dysregulated immune system and increased gene expression associated with SARS-CoV-2 susceptibility may at least partially account for COVID-19 vulnerability in the elderly.
Adult ; Aged ; Aged, 80 and over ; Aging ; genetics ; immunology ; Betacoronavirus ; CD4-Positive T-Lymphocytes ; metabolism ; Cell Lineage ; Chromatin Assembly and Disassembly ; Coronavirus Infections ; immunology ; Cytokine Release Syndrome ; etiology ; immunology ; Cytokines ; biosynthesis ; genetics ; Disease Susceptibility ; Flow Cytometry ; methods ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gene Rearrangement ; Humans ; Immune System ; cytology ; growth & development ; immunology ; Immunocompetence ; genetics ; Inflammation ; genetics ; immunology ; Mass Spectrometry ; methods ; Middle Aged ; Pandemics ; Pneumonia, Viral ; immunology ; Sequence Analysis, RNA ; Single-Cell Analysis ; Transcriptome ; Young Adult