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Acute kidney injury (AKI) is characterized by necroinflammation formed by necrotic tubular epithelial cells (TECs) and interstitial inflammation. In necroinflammation, macrophages are key inflammatory cells and can be activated and polarized into proinflammatory macrophages. Membranous Toll-like receptors (TLRs) can cooperate with intracellular NOD-like receptor protein 3 (NLRP3) to recognize danger signals from necrotic TECs and activate proinflammatory macrophages by assembling NLRP3 inflammasome. However, the cooperation between TLRs and NLRP3 is still unclear. Using conditioned medium from necrotic TECs, we confirmed that necrotic TECs could release danger signals to activate NLRP3 inflammasome in macrophages. We further identified that necrotic TECs-induced NLRP3 inflammasome activation was dependent on ATP secretion via Pannexin-1 (Panx1) channel in macrophages. Next, we verified that TLR2 was required for the activation of Panx1 and NLRP3 in macrophages. Mechanistically, we indicated that caspase-5 mediated TLR2-induced Panx1 activation. In addition, we showed that necrotic TECs-induced activation of TLR2/caspase-5/Panx1 axis could be decreased in macrophages when TECs was protected by N-acetylcysteine (NAC). Overall, we demonstrate that danger signals from necrotic TECs could activate NLRP3 inflammasome in macrophages via TLR2/caspase-5/Panx1 axis during AKI.

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Copper pipeline is a commonly used industrial transmission pipeline. Nondestructive testing of copper pipeline early damage is very important. Laser scanning has attracted extensive attention because it can realize the visualization of guided wave propagation and non-contact on-line detection. However, the damage points detection in laser scanning imaging method rely on the difference between the damage points signals and surrounding normal points signals. This limits the applicability of laser scanning and may lead to inaccurate in large-area detection. Facing with such challenges, a damage detection method based on CNN-LSTM network is proposed for laser ultrasonic guided wave scanning detection in this paper, which can detect each scanning point signal without relying on the surrounding detection points signals. Firstly, the proposed data conversion algorithm is used to preprocess the laser scanning signals. Next, CNN-LSTM network is used to train the damage detection model. Four 1D Conv channels with different convolution kernel sizes and depths are designed in Convolutional Neural Network (CNN) module. The module can extract the signal time domain features. Then the features are input into the Long Short-Term Memory Network (LSTM) for feature extraction and classification. Finally, the CNN-LSTM is trained using the laser scanning detection data collected on the copper pipeline with crack and corrosion damages, and applied to detect the copper pipeline damage signal. At the same time, the state-of-the-art methods is compared with proposed method. The experimental results show that the detection accuracy of the method is 99.9%, 99.9%, 99.8% and 99.8% for copper pipeline 0.5 mm deep crack damage, penetrating crack damage, corrosion damage and inside crack damage, respectively. The damage location and size can be accurately detected by the proposed method.

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This meta-analysis aimed to compare the clinical and radiographic outcomes between mobile-bearing total knee arthroplasty (MB-TKA) and fixed-bearing total knee arthroplasty (FB-TKA) at a minimum 10-year follow-up. PubMed, EMBASE, and Cochrane databases were searched. All included articles were evaluated by two trained reviewers according to the guidelines of the Cochrane Collaboration Handbook for potential risk, and the Consolidated Standards on Reporting Trials (CONSORT) checklist and scoring system was also used to assess the methodological quality of each study. The extracted data included function scores, range of motion (ROM) of the knee, incidence of adverse events or revision, survivorship analysis, and radiographic outcomes. Seven randomized controlled trials (RCTs) were included in this meta-analysis, and all RCTs had a follow-up period longer than 10 years. This meta-analysis shows no significant difference between the two groups with respect to the Keen Society Score (KSS; p = 0.38), KSS function score (p = 0.30), the Western Ontario and McMaster Universities Osteoarthritis index (WOMAC; p = 0.59), ROM (p = 0.71), radiolucent line (p = 0.45), femoral and tibial component positions in the coronal plane (p = 0.55 and 0.35, respectively), revision incidence (p = 0.77), and survivorship rates (p = 0.39). Meanwhile, it showed a slight difference between the two groups in the tibial component position in the sagittal plane (p = 0.003). According to this meta-analysis, the current best available evidence suggests no significant difference between the MB-TKA and FB-TKA groups with respect to the clinical outcomes, radiographic outcomes, revision, and survivorship at a minimum 10-year follow-up. This is a Level II, meta-analysis study.

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Acute kidney injury (AKI) is a common clinical problem, and patients who survive AKI have a high risk of chronic kidney disease (CKD). The mechanism of CKD post-AKI, characterized by progressive renal fibrosis, is still unclear. Maladaptive tubular epithelial cells (TECs) after AKI are considered a leading cause of renal fibrosis post-AKI. TECs under maladaptive repair manifest characteristics of senescence. Removing senescent TECs by genetic ablation has been proven effective in reducing renal fibrosis. Senolytics, which eliminate senescent cells by pharmacological intervention, have been studied in a series of degenerative diseases. To our knowledge, the effects of senolytics on renal fibrosis post-AKI have not been verified before. Here, we confirmed renal senescence in the unilateral ischemia/reperfusion injury murine model. Senescent TECs could activate fibroblasts and senolytics specifically induced apoptosis of senescent TECs. Next, we demonstrated that senolytics could reduce renal senescence and ameliorate renal fibrosis in both unilateral renal ischemia/reperfusion injury and multiple-cisplatin-treatment murine models. Our results indicate senescent TECs as a vital factor in renal fibrosis progression, and senolytic therapy might be promising for treating CKD post-AKI.

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OBJECTIVE: To investigate the effect of palbociclib on cell cycle progression and proliferation of human renal tubular epithelial cells. METHODS: Human renal tubular epithelial cell line HK-2 was treated with 1, 5, 10, and 20 µmol/L of palbociclib, and the changes in cell proliferation and viability were examined by cell counting and CCK8 assay. EDU staining was used to assess the proliferation of HK-2 cells following palbiciclib treatment at different concentrations for 5 days. The effect of palbociclib on cell cycle distribution of HK-2 cells was evaluated using flow cytometry. SA-ß-Gal staining and C12FDG senescence staining were used to detect senescence phenotypes of HK-2 cells after palbociclib treatment at different concentrations for 5 days. The relative mRNA expression levels of P16, P21, and P53 and the genes associated with senescence-related secretion phenotypes were detected by RT-PCR, and the protein expressions of P16, P21 and P53 were detected by Western blotting. RESULTS: Palbociclib inhibited HK-2 cell proliferation and induced cell cycle arrest in G1 phase. Compared with the control cells, HK-2 cells treated with high-dose (10 µmol/L) palbociclib exhibited significantly suppressed cell proliferation activity, and the inhibitory effect was the most obvious on day 5 (P < 0.01). Palbociclib treatment significantly reduced the number of cells in S phase (P < 0.01) and induced senescence of HK-2 cells. The results of SA-ß-Gal and C12FDG senescence staining showed a significantly enhanced activity of intracellular senescence-related galactosidase in palbociclib-treated HK-2 cells, suggesting significant senescence of the cells (P < 0.01). RT-PCR and Western blotting showed that palbociclib treatment significantly increased the mRNA and protein expression levels of P16, P21, and P53 in HK-2 cells (P < 0.01); the mRNA expression levels of senescence-related secretory factors also increased significantly in HK-2 cells after palbociclib treatment (P < 0.01). CONCLUSIONS: Palbociclib induces HK-2 cell senescence by causing cell growth arrest and delaying cell cycle progression.

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Osteoarthritis (OA), as one of the top 10 causes of physical disability, is characterized by inflammation of the synovial membrane and progressive destruction of the articular cartilage. Cinnamic aldehyde (CA), an α,ß-unsaturated aldehyde extracted from the traditional Chinese herbal medicine cinnamon (Cinnamomum verum J.Presl), has been reported to have anti-inflammatory, antioxidant, and anticancer properties. However, the anti-inflammatory effect of CA on OA remains unclear. The purpose of the present study was to investigate the effects of CA on inflammation, and cartilage degeneration in OA. A CCK-8 assay was performed to assess the potential toxicity of CA on cultured human OA chondrocytes. Following treatment with lipopolysaccharide (LPS) and CA, the expression of proinflammatory cytokines, including interleukin (IL)-1ß, IL-6, and tumor necrosis factor-alfa (TNF-α), was evaluated using quantitative real-time polymerase chain reaction (RT-qPCR) analysis, enzyme-linked immunosorbent assay, and Western blotting (WB). The production of matrix metalloproteinase-13 (MMP-13) and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5) was also examined using RT-qPCR and WB. Furthermore, to investigate the potential anti-inflammatory mechanism of CA, biomarkers of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway (p65, IKB-α) were detected using WB. The results demonstrated that CA significantly inhibited the expressions of IL-1ß, IL-6, TNF-α, MMP-13, and ADAMTS-5 in LPS-induced OA chondrocytes. CA dramatically suppressed LPS-stimulated NF-κB activation. Collectively, these results suggest that CA treatment may effectively prevent OA.

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OBJECTIVE: The aim of this study is to compare the efficiency of different separation techniques for extracting synovial tissue-derived exosomes. METHODS: The synovial tissue discarded during knee arthroscopy or total knee arthroplasty surgery was collected from the Third Affiliated Hospital of Beijing University of Chinese Medicine. Ultracentrifugation (UC), filtration combined with size exclusion chromatography (SECF), and 8% polyethylene glycol (PEG) were used to extract synovial tissue-derived exosomes. Transmission electron microscopy (TEM), nanoparticle tracer analysis (NTA), and Western Blot (WB) were used to detect the morphology, particle size, and biomarker proteins (CD9, CD63, Flotillin-1, and calnexin) of exosomes. RESULTS: The extracts of enriched round and discoid vesicles were successfully extracted with UC, SECF, and PEG. The results of TEM have shown that all three extraction methods can extract circular or elliptical vesicles with disc- and cup-shaped structures from the synovial tissue, with the diameter is about 30-150 nm. NTA suggested the main peaks of three groups of exosomes are around 100-120 nm, and the concentration of the three groups of exosomes was greater than 1 × 1010/ml. The results of WB showed that three positive protein markers (CD9, CD63, and Flotillin-1) were highly expressed in the suspension extracted by the three methods and low in the synovial tissue. However, the negative protein (calnexin) was highly expressed in synovial tissues and PEG group, while low in UC and SECF group. CONCLUSION: Morphology, particle size, and labeled protein marker detection confirmed that UC, SECF, and PEG can extract exosomes derived from synovial tissue; UC and SECF are more recommended for the extraction of synovial tissue-derived exosomes, which provides a methodological basis for studying the function and mechanism of synovial tissue exosomes in the future.

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OBJECTIVE: The purpose of this study was (1) to perform a summary of meta-analyses comparing platelet-rich plasma (PRP) injection with hyaluronic acid (HA) and placebo injection for KOA patients, (2) to determine which meta-analysis provides the best available evidence to making proposals for the use of PRP in the treatment of KOA patients, and (3) to highlight gaps in the literature that require future investigation. MATERIAL AND METHODS: PubMed, EMBASE, and Cochrane databases search were performed for meta-analyses which compared PRP injection with HA or placebo. Clinical outcomes and adverse events were extracted from these meta-analyses. Meta-analysis quality was assessed using the Quality of Reporting of Meta-analyses (QUOROM) systems and the Oxman-Guyatt quality appraisal tool. The Jadad decision algorithm was also used to determine which meta-analysis provided the best available evidence. RESULTS: Four meta-analyses were included in our study, and all of these articles were Level I evidence. The QUOROM score of each included meta-analysis range from 14 to 17 points (mean score 15, maximum score 18), and the Oxman-Guyatt score range from 4 to 6 points (mean score 5, maximum score 7). Three meta-analyses indicated PRP showed more benefit in pain relief and functional improvement than the control group, and the other one suggested no difference between these groups. All included meta-analyses found no statistical difference in adverse events between these groups. In addition, a meta-analysis conducted by Shen et al. got the highest methodological quality score and suggested that PRP provided better pain relief and function improvement in the treatment of KOA. CONCLUSIONS: For short-term follow-up (≤1 year), intra-articular PRP injection is more effective in terms of pain relief and function improvement in the treatment of KOA patients than HA and placebo, and there is no difference in the risk of an adverse event between PRP and HA or placebo. LEVEL OF EVIDENCE: Level I evidence, a summary of meta-analyses TRIAL REGISTRATION: PROSPERO ID CRD42018116168.

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OBJECTIVE: To investigate the effect of blocking pannexin-1 against acute kidney injury induced by cisplatin. METHODS: Twenty-six male C57BL/6 mice aged 6-8 weeks were randomly divided into control group, cisplatin model (Cis) group and cisplatin + carbenoxolone treatment group (Cis + CBX). In Cis group and Cis + CBX group, the mice were injected intraperitoneally with 20 mg/kg of cisplatin and with CBX (20 mg/kg) at 30 min before and 24 and 48 h after cisplatin inhjection, respectively. All the mice were sacrificed at 72 h after cisplatin injection, and plasma and kidney samples were collected for testing mRNA and protein expression levels of pannexin-1 in the renal tissue using RT-qPCR and Western blotting and for detecting plasma creatinine and BUN levels; the pathological changes in the renal tissues were observed using Periodic Acid-Schiff staining. The expression of kidney injury molecule 1 (KIM-1) was examined using immunohistochemistry and the mRNA expressions of KIM-1 and neutrophil gelatinase- related lipid transport protein (NGAL) were detected by RT-qPCR to evaluate the injuries of the renal tubules. The infiltration of F4/80-positive macrophages and CD4-positive T cells were observed by immunofluorescence. In the in vitro experiment, human proximal tubule epithelial cell line HK-2 was stimulated with 50 µmol/L cisplatin to establish a cell model of acute kidney injury, and the mRNA and protein expressions of pannexin-1 were detected by RT-qPCR and Western blotting at 4, 6, 12, 18 and 24 h after the stimulation. RESULTS: Compared with the control mice, the cisplatin-treated mice showed significantly up-regulated protein levels (P < 0.05) and mRNA levels (P < 0.005) of pannexin-1 in the kidney tissue. Cisplatin stimulation also caused significant increases in the protein levels (P < 0.005) and mRNA levels (P < 0.005) of pannexin-1 in cultured HK-2 cells. Compared with cisplatin-treated mice, the mice treated with both cisplatin and the pannexin-1 inhibitor CBX showed obviously lessened kidney pathologies and milder renal tubular injuries with significantly reduced plasma BUN and Scr levels (P < 0.01), expressions of KIM-1 and NGAL in the kidney (P < 0.05), and infiltration of F4/80-positive macrophages (P < 0.01) and CD4- positive T cells (P < 0.05) in the kidney tissues. CONCLUSIONS: In cisplatin induced acute kidney injury mice model, Pannexin-1 expression is up-regulated in the kidneys tissue, and blocking pannexin-1 alleviates the acute kidney injury via reducing renal inflammatory cell infiltration.

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Optimal sensor placement is a significant task for structural health monitoring (SHM). In this paper, an SHM system is designed which can recognize the different impact location and impact degree in the composite plate. Firstly, the finite element method is used to simulate the impact, extracting numerical signals of the structure, and the wavelet decomposition is used to extract the band energy. Meanwhile, principal component analysis (PCA) is used to reduce the dimensions of the vibration signal. Following this, the non-dominated sorting genetic algorithm (NSGA-II) is used to optimize the placement of sensors. Finally, the experimental system is established, and the Product-based Neural Network is used to recognize different impact categories. Three sets of experiments are carried out to verify the optimal results. When three sensors are applied, the average accuracy of the impact recognition is 59.14%; when the number of sensors is four, the average accuracy of impact recognition is 76.95%.