AstudyofPt-metabolitesfromanewanti-hepatomadrugmiriplatinwasimportanttomiriplatin's pharmacology research. Therefore, a method based on size exclusion chromatography-inductively coupled plasma mass spectrometry ( SEC-ICP-MS) was developed to study miriplatin and its Pt-metabolites in Beagle dog plasma. This method could be used to study total platinum concentration half-quantitatively. Compared with traditional ICP-MS direct determination, data acquired from this SEC-ICP-MS method were almost the same. By using BioSep-s2000 column, 25 mmol/L of pH 7. 2 phosphate buffer as eluent, and Pt-195 as detecting isotope, we discovered miriplatin with its four Pt-metabolites in dog plasma after intra-hepatic artery administration. The main Pt-metabolite was m2 , which associated with plasma proteins. Miriplatin in plasma did not bind with plasma proteins. According to calculation, the ratio of miriplatin/m2 first decreased rapidly, and then slowly increased to its second climax, finally slowly decreased.

Cytoplasmic processing bodies, termed P bodies, are involved in diverse post-transcriptional processes including mRNA decay, nonsense-mediated RNA decay (NMD), RNAi, miRNA-mediated translational repression and storage of translationally silenced mRNAs. Regulation of the formation of P bodies in the context of multicellular organisms is poorly understood. Here we describe a systematic RNAi screen in C. elegans that identified 224 genes with diverse cellular functions whose inactivations result in a dramatic increase in the number of P bodies. 83 of these genes form a complex functional interaction network regulating NMD. We demonstrate that NMD interfaces with many cellular processes including translation, ubiquitin-mediated protein degradation, intracellular trafficking and cytoskeleton structure.We also uncover an extensive link between translation and RNAi, with different steps in protein synthesis appearing to have distinct effects on RNAi efficiency. Moreover, the intracellular vesicular trafficking network plays an important role in the regulation of RNAi. A subset of genes enhancing P body formation also regulate the formation of stress granules in C. elegans. Our study offers insights into the cellular mechanisms that regulate the formation of P bodies and also provides a framework for system-level understanding of NMD and RNAi in the context of the development of multicellular organisms.
Animals ; Animals, Genetically Modified ; Caenorhabditis elegans ; genetics ; Cytoplasmic Structures ; Gene Expression Regulation ; Genes, Helminth ; Genome, Helminth ; genetics ; MicroRNAs ; genetics ; Nonsense Mediated mRNA Decay ; physiology ; RNA Interference ; RNA, Helminth ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective:To explore the correlation between fibular head height and varus knee osteoarthritis occurrence and severity.Methods:A retrospective analysis was performed on 618 participants (618 knees, 184 males and 434 females, mean age 61.12±10.98 years) who underwent standard weight-bearing full-leg radiographs and were diagnosed as non-knee osteoarthritis or varus knee osteoarthritis from January 2019 to June 2019. Knee osteoarthritis was diagnosed according to Kellgren-Lawrence grading: 0-I grades were diagnosed as non-osteoarthritis, II-IV grades were diagnosed as osteoarthritis. Joint line convergence angle (JLCA), medial proximal tibial angle (MPTA) and hip-knee-ankle angle were measured on X-rays to reflect varus deformity. The fibular head height was defined as the vertical distance from upper edge of fibular head to lateral tibial plateau. Patients were divided into 5 groups according to Kellgren-Lawrence grading. Differences of age, gender, height, weight, body mass index, varus deformity (JLCA, MPTA and hip-knee-ankle angle) between Kellgren-Lawrence 0-IV grades were compared. Ordinal logistic regression was performed to analyze the correlation between fibular head height and Kellgren-Lawrence grades. Pearson's correlation analysis was used for the correlation among fibular head height, JLCA, MPTA and hip-knee-ankle angle, and the main factor of JLCA, MPTA and hip-knee-ankle angle was extracted by factor analysis. Multiple linear regressions were used to analyze the correlation between fibular head height and varus deformity score.Results:There were 68, 66, 97, 98, 289 participants in Kellgren-Lawrence grades 0-IV respectively that was 134 participants were diagnosed as non-osteoarthritis and 484 participants were diagnosed as osteoarthritis. Fibular head height and MPTA showed a decreasing trend ( F=129.076, 24.875; P<0.001) while JLCA and hip-knee-ankle angle showed an increasing trend ( F=414.346, 105.996; P<0.001) with the increase in Kellgren-Lawrence grading. Age, body mass index and fibular head height are influencing factors of Kellgren-Lawrence grading with OR(95%CI) were 1.116(1.093, 1.141), 1.363(1.060, 1.754), 0.617(0.575, 0.662) . Fibular head height was negatively correlated with JLCA and hip-knee-ankle angle ( r=-0.641, -0.478; P<0.001) , respectively, and positively correlated with MPTA ( r=0.320, P<0.001). There were significant correlations between age, fibular head height and the varus deformity score ( β=0.274, -0.457; P<0.001). Conclusion:Fibular head height of patients with varus knee osteoarthritis is lower than that of non-osteoarthritis. In addition to age and body mass index, fibular head height is a risk factor for varus knee osteoarthritis occurrence. The smaller the fibular head height is, the more serious the osteoarthritis severity and varus deformity are.

Objective:To analyze the risk factors for rotator cuff tear and evaluate the corresponding prediction efficacy.Methods:A case-control study was conducted in 69 patients with rotator cuff tear admitted to Affiliated Hospital of Qingdao University from June 2020 to June 2021 (rotator cuff tear group) and 51 normal volunteers or medical examiners (normal control group). There were 55 males and 65 females, with the age range of 34-77 years [(58.2±7.2)years]. Body mass index (BMI) ranged from 19.5-32.4 kg/m 2 [(25.4±2.5)kg/m 2]. Univariate analysis was performed for the correlation of gender, age, history of hypertension, history of smoking, history of diabetes, BMI, angle of humeral greater tuberosity and notch angle of humeral greater tuberosity with rotator cuff tear in the two groups. Factors with statistically significant differences were included in multivariate Logistic regression analysis to determine the independent risk factors for rotator cuff tear. Spearman correlation analysis was used to find factor correlation between the two groups. The receiver operating characteristic (ROC) curve of relevant factors for the diagnosis and prediction of rotator cuff tear was analyzed. Results:Univariate analysis showed that age, angle of humeral greater tuberosity and notch angle of humeral greater tuberosity related to rotator cuff tear (all P<0.05). On the contrary, gender, history of hypertension, history of smoking, history of diabetes and BMI were not correlated with rotator cuff tear (all P>0.05). Multivariate Logistic regression analysis showed significant correlations of angle of humeral greater tuberosity and notch angle of humeral greater tuberosity with rotator cuff tear (all P<0.05). Spearman correlation analysis showed that age was not correlated with angle of humeral greater tuberosity and notch angle of humeral greater tuberosity ( r=0.09, 0.13, all P>0.05), but there was significant positive correlation between angle of humeral greater tuberosity and notch angle of humeral greater tuberosity ( r=0.76, P<0.01). When the optimal cutoff values of angle of humeral greater tuberosity and notch angle of humeral greater tuberosity were 70.05° and 150.55°, the area under the ROC curve (AUC) for predicting rotator cuff tear was 0.79 (95% CI 0.71-0.87, P<0.01) and 0.81 (95% CI 0.74- 0.89, P<0.01). Conclusions:Angle of humeral greater tuberosity and notch angle of humeral greater tuberosity are independent factors for rotator cuff tear, and larger angles suggest higher prevalence of rotator cuff tear. Besides, the notch angle of humeral greater tuberosity has relatively better predictive performance.

Objective:To compare the clinical efficacy between the pressing tendon suture with a lateral anchor and the knotted suture with a lateral anchor in the treatment of severe rotator cuff tear under shoulder arthroscopy.Methods:A retrospective study was conducted of the 42 patients who had been treated at Department of Orthopedic Sports Medicine, Affiliated Hospital to Qingdao University from December 2018 to December 2019 for severe rotator cuff tear. They were 13 males and 29 females, with an age of (58.3±10.1) years (from 32 to 74 years). The injury was on the right side in 26 cases and on the left side in 16 cases. Of them, 22 received pressing tendon suture with a lateral anchor under shoulder arthroscopy and 20 knotted suture with a lateral anchor under shoulder arthroscopy. The flexion, abduction and external rotation of the shoulder, Visual Analog Scale (VAS) pain score, University of California-Los Angeles (UCLA) score, Constant-Murley shoulder score, American Shoulder and Elbow Surgeons (ASES) score and imaging MRI Sugaya grading were compared between the 2 groups 12 months after operation.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability between them ( P>0.05). At 12 months after operation, the pressing tendon suture group had better abduction and flexion of the shoulder, a lower VAS pain score and a higher ASES score than the knotted suture group, but the differences were insignificant ( P>0.05). At 12 months after operation, the pressing tendon suture group achieved significantly better external rotation (39.2°±11.7°) and significantly higher UCLA score (28.1±4.7) and Constant-Murley shoulder score (77.0±9.3) than the knotted suture group (29.8°±14.6°, 22.1±5.7 and 66.4±11.0) ( P<0.05). At 12 months after operation, the imaging MRI Sugaya grading was significantly lower for the pressing tendon suture group than that for the knotted suture group ( P<0.05). Conclusion:In the treatment of patients with severe rotator cuff tear under shoulder arthroscopy, the pressing tendon suture with a lateral anchor may lead to better clinical prognosis than the knotted suture with a lateral anchor.

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

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins