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Jingxia Sun, Jianhao Huang, Wensheng Lu Department of Endocrinology and Metabolism, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

Jiaxia Han, Wensheng Lu Department of Endocrinology, People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

Qiu Wang, Xiuxian Huang, Wensheng Lu Department of Endocrinology and Metabolism, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

A process for photocatalytically synthesizing hydrogen peroxide while simultaneously degrading organic pollutants is of interest. In this study, a dendritic nanopolymer was synthesized and utilized as a photocatalyst for the production of hydrogen peroxide, which was then used in tandem with the degradation of Tetracycline (TC) and Rhodamine B (Rh B) organic pollutants under a 36 W incandescent bulb. The yield of H2O2 was measured at 8.4 mg g−1 h−1, with photocatalytic degradation rates of 93.90 % for Rh B and 72.92 % for TC. The degradation kinetics of TC and Rh B by the dendritic polymer were found to follow pseudo‐first‐order kinetics, and a higher affinity for degradation was observed for Rh B. The photocatalytic degradation mechanism of TC in a Fenton‐like system was attributed to an oxidation system involving hydrogen peroxide and ferrous ions. [ABSTRACT FROM AUTHOR]

Wenjun Lu,1,* Quedan Qiu,2,* Keda Chen,2 Rongqing Zhao,2 Qingcao Li,2 Qiaoping Wu2 1Intensive Care Units of Ningbo Medical Centre Lihuili Hospital, Ningbo University, Ningbo, Zhejiang Province, People’s Republic of China; 2Clinical Laboratory of Ningbo Medical Centre Lihuili Hospital, Ningbo University, Ningbo, Zhejiang Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Qingcao Li; Qiaoping Wu, Tel +86-574-55835786, Fax +86-574-55835781, Email lqc_lab@163.com; lhlyywqp@163.comBackground: Integrons are the main mode of horizontal transmission of drug-resistance genes and are closely related to drug resistance in clinical bacteria. In this study, the distributions of class 1, 2, and 3 integron gene cassettes were investigated in 150 Proteus mirabilis (P. mirabilis) isolates from patients, and molecular characterization of functional class 2 integrons was further analyzed.Methods: Class 1, 2, and 3 integrons were screened by polymerase chain reaction (PCR) in 150 clinical P. mirabilis isolates. The variable regions of the integrons were determined by restriction analysis and sequencing. Internal stop codons mutations in class 2 integrons and their common promoters were also determined by sequencing. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) was used to analyze the phylogenetic relations of class 2 integron-positive isolates.Results: Class 1 integrons were detected in 69 (46%) of 150 P. mirabilis isolates, and six different gene cassette arrays were detected, with the most prevalent being dfrA32-aadA2. Class 2 integrons were detected in 61 (40.7%) of 150 P. mirabilis isolates, and three different gene cassette arrays were detected, including sat2-aadA1, which was detected for the first time in a class 2 integron. Nearly similar ERIC-PCR fingerprinting patterns were detected in 45 (73.8%) of 61 class 2 integron-positive isolates. The functional class 2 integron was detected in three P. mirabilis isolates having the same gene cassette, dfrA1-sat2-aadA1, in the variable region and four novel open reading frames with unknown functions. Same PintI2 and Pc promoters were detected in these three functional class 2 integron isolates, as was found in other class 2 integron isolates. However, these three strains did not totally show identical homology and drug sensitivity.Conclusion: Although functional class 2 integrons have low distribution and relatively conserved molecular characteristics, they can still form clinical dissemination and drug resistance expression.Keywords: Proteus mirabilis, integrons, premature stop codon, promoter regions, antibacterial drug resistance

Na Wang,1 Minghua Zhan,1,2 Jianhua Liu,3 Yao Wang,4 Yongwang Hou,5 Caiqing Li,1 Jia Li,1 Xuying Han,1 Jinlu Liu,1 Yong Chen,6 Jingjing Fan,6 Jianhua Tang,7 Wenhua Lu,8 Xinran Zhong,1 Zhihua Zhang,3 Wei Zhang1,4 1Microbiology Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 2Clinical Laboratory, Peking University People’s Hospital, Beijing, 100730, People’s Republic of China; 3Respiratory Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 4Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 5Clinical Laboratory, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 6Infectious Disease Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 7Clinical Pharmacy Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 8Dental Department, Beijing Fengtai Tieying Community Health Service Center, Beijing, 100730, People’s Republic of ChinaCorrespondence: Wei Zhang; Zhihua Zhang, Tel +86-15369318318, Email 15369318318@163.com; zzh19641229@163.comPurpose: In this study, we analyzed the clinical distribution and drug resistance of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains, the minimum inhibitory concentrations (MIC), MIC50 and MIC90, and geographical distribution in Hebei Province, China. We aimed to provide epidemiological research data, formulate appropriate combined treatment schemes, reasonably select antibiotics, and standardize nosocomial infection control schemes.Patients and Methods: A total of 6328 strains of CRKP were collected from 2017 to 2019. The MIC was determined for the drug sensitivity test, and the experimental data were statistically analyzed using WHONET5.6.Results: The detection rate of CRKP increased annually from 13.4% in 2017 to 14.5% in 2018, and 14.6% in 2019. The ratio of males to females was approximately 2:1; 53.6% were elderly, 39% were adults, 4.8% were minors, and 2.5% were newborns. The specimens collected were mainly sputum (70.9%). The resistance rate of CRKP to carbapenems and other β-lactam antibiotics was found to be increasing, with resistance rates generally greater than 90%. The resistance rate to aminoglycoside antibiotics decreased yearly to approximately 50%, and the resistance rate to quinolones remained unchanged at approximately 80%. From 2017 to 2019, the resistance rate of CRKP in Hebei Province to various antibiotics was high, and the resistance rate to β-lactam antibiotics increased each year.Conclusion: The situation of CRKP resistance is severe in Hebei Province, China. The resistance rate to most antibiotics is very high and shows an upward trend. Among them, the resistance rate to polymyxin is low; however, few resistant strains do exist. MIC50 and MIC90 are higher than their MICs. It mainly causes lung infection in elderly men. This study is helpful to improve the diagnosis, treatment, and prevention of CRKP infection in our province.Keywords: carbapenems, clinical distribution, drug resistance, minimum inhibitory concentration, geographic distribution, Klebsiella pneumoniae

Wang Li,1,2,* Mei Mei,1,3– 5,* Tian Liu,1,* ShuWen Zhang,1 ZeYu Wang,1 YuHong Suo,1 Shuai Wang,1 Yang Liu,1 NingNing Zhang,1 Wei Lu1 1Department of Hepatobiliary Oncology, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University, Tianjin, 300060, People’s Republic of China; 2NHC Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin, 300134, People’s Republic of China; 3Graduate School of Tianjin Medical University, Tianjin, 300070, People’s Republic of China; 4Tianjin First Central Hospital, Tianjin, 300192, People’s Republic of China; 5Department of Gastroenterology, Tianjin Haihe Hospital, Tianjin, 300350, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wei Lu; NingNing ZhangDepartment of Hepatobiliary Oncology, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University, Tianjin, 300060, People’s Republic of ChinaEmail mail4luwei@163.com; mail4ningning@163.comBackground: Hepatocellular carcinoma has been identified to be among the most prevalent malignancies in the world and has an unfavorable prognosis. Immune checkpoints perform an essential function in many biological processes and are associated with the survival of cancer patients. The function of immune checkpoints remains unknown.Methods: We used bioinformatic methods to examine the prognostic value of immune checkpoints and the corresponding link to immune infiltration in HCC. qRT-PCR was used to validate the expression of immune checkpoints and their prognostic significance in HCC.Results: The level of mRNA of SIGLEC15, PDCD1LG2, LAG3, PDCD1, CTLA4 as well as PDCD1LG2 was increased in HCC tissues as opposed to liver tissues. Immune checkpoints were shown to participate in the activation of the apoptotic pathway in HCC patients. The elevated expression of PDCD1 and PDCD1LG2 were shown to have a favorable recurrence-free survival (RFS), progression-free survival (PFS), disease-specific survival (DSS), and overall survival (OS). PDCD1, PDCD1LG2, and pT stage were independent variables that affect the HCC patients’ prognoses as revealed by the multivariate and univariate analyses. A prediction nomogram indicated that the calibration plots for OS rates over three and five years had a stronger predictive performance in the TCGA HCC cohort in contrast with an ideal model. Positive correlations were observed between the PDCD1 and PDCD1LG2 expression and immune biomarkers, immune cells, chemokine receptors, as well as chemokines.Conclusion: The present research performed a thorough examination of the prognostic significance of immune checkpoints in HCC and its correlation with immune infiltration, which suggested that PDCD1 and PDCD1LG2 were prognostic biomarkers in HCC and related to the immune infiltration.Keywords: HCC, PDCD1, PDCD1LG2, immune checkpoints, prognosis biomarker

Samuel James Brake,1 Mathew Suji Eapen,1 Kielan Darcy McAlinden,1 James Markos,1,2 Greg Haug,1,2 Josie Larby,1,2 Collin Chia,1,2 Ashutosh Hardikar,1,3 Gurpreet Kaur Singhera,4,5 Tillie L Hackett,4,5 Wenying Lu,1 Sukhwinder Singh Sohal1 1Respiratory Translational Research Group, Department of Laboratory Medicine, School of Health Sciences, College of Health and Medicine, University of Tasmania, Launceston, TAS, 7248, Australia; 2Department of Respiratory Medicine, Launceston General Hospital, Launceston, TAS, 7250, Australia; 3Department of Cardiothoracic Surgery, Royal Hobart Hospital, Hobart, TAS, 7000, Australia; 4Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, BC, Canada; 5Department of Medicine, University of British Columbia Centre for Heart Lung Innovation, St. Paul’s Hospital, Vancouver, BC, CanadaCorrespondence: Sukhwinder Singh SohalRespiratory Translational Research Group, Department of Laboratory Medicine, School of Health Sciences, College of Health and Medicine, University of Tasmania, Locked Bag – 1322, Newnham Drive, Launceston, TAS, 7248, AustraliaTel +61 3 6324 5434Email sssohal@utas.edu.auBackground: Smokers and patients with COPD are highly susceptible to SARS-CoV-2 infection, leading to severe COVID-19.Methods: This cross-sectional study involved resected lung tissues from 16 patients with GOLD stage I or II COPD; of which 8 were current smokers COPD (COPD-CS), and 8 ex-smokers COPD (COPD-ES), 7 normal lung function smokers (NLFS), 9 patients with small airways disease (SAD), and 10 were never-smoking normal controls (NC). Immunostaining for ACE2, Furin, and TMPRSS2 was performed and analysed for percent expression in small airway epithelium (SAE) and counts for positively and negatively stained type 2 pneumocytes and alveolar macrophages (AMs) were done using Image ProPlus V7.0. Furthermore, primary small airway epithelial cells (pSAEC) were analysed by immunofluorescence after exposure to cigarette smoke extract (CSE).Results: ACE2, Furin, and TMPRSS2 expression significantly increased in SAE and type 2 pneumocytes in all the subjects (except Furin for NLFS) compared to NC (p < 0.001). Similar significance was observed for ACE2 positive AM (p < 0.002), except COPD-ES, which decreased in ACE2 positive AMs (p < 0.003). Total type 2 pneumocytes and AMs significantly increased in the pathological groups compared to NC (p < 0.01), except SAD (p = 0.08). However, AMs are significantly reduced in COPD-ES (p < 0.003). Significant changes were observed for tissue co-expression of Furin and TMPRSS2 with ACE2 in SAE, type 2 pneumocytes and AMs. These markers also negatively correlated with lung function parameters, such as FEV1/FVC % predicted, FEF25-75%, DLCO% predicted. A strong co-localisation and expression for ACE2 (p < 0.0001), Furin (p < 0.01), and TMPRSS2 (p < 0.0001) was observed in pSAEC treated with 1% CSE than controls.Discussion: The increased expression of ACE2, TMPRSS2 and Furin, in the SAE, type 2 pneumocytes and AMs of smokers and COPD are detrimental to lung function and proves that these patient groups could be more susceptible to severe COVID-19 infection. Increased type 2 pneumocytes suggest that these patients are vulnerable to developing post-COVID-19 interstitial pulmonary fibrosis or fibrosis in general. There could be a silently developing interstitial pathology in smokers and patients with COPD. This is the first comprehensive study to report such changes.Keywords: COVID-19, SARS-CoV-2, smoking, COPD, ACE2, type 2 pneumocytes, alveolar macrophages, epithelium

Midbrain dopamine neurons are crucial for many behavioral and cognitive functions. As the major excitatory input, glutamatergic afferents are important for control of the activity and plasticity of dopamine neurons. However, the role of glutamatergic input as a whole onto dopamine neurons remains unclear. Here we developed a mouse line in which glutamatergic inputs onto dopamine neurons are specifically impaired, and utilized this genetic model to directly test the role of glutamatergic inputs in dopamine-related functions. We found that while motor coordination and reward learning were largely unchanged, these animals showed prominent deficits in effort-related behavioral tasks. These results provide genetic evidence that glutamatergic transmission onto dopaminergic neurons underlies incentive motivation, a willingness to exert high levels of effort to obtain reinforcers, and have important implications for understanding the normal function of the midbrain dopamine system.

The development of superconducting ECR source with higher magnetic fields and higher microwave frequency is the most straight forward path to achieve higher beam intensity and higher charge state performance. SECRAL, a superconducting third generation ECR ion source, is designed for 24-28 GHz microwave frequency operation with an innovative magnet configuration of sextupole coils located outside the three solenoids. SECRAL at 24 GHz has already produced a number of record beam intensities, such as 40Ar12+ 1.4 emA, 129Xe26+ 1.1 emA, 129Xe30+ 0.36 emA, and 209Bi31+ 0.68 emA. SECRAL-II, an upgraded version of SECRAL, was built successfully in less than 3 years and has recently been commissioned at full power of a 28 GHz gyrotron and three-frequency heating (28 + 45 + 18 GHz). New record beam intensities for highly charged ion production have been achieved, such as 620 eμA 40Ar16+, 15 eμA 40Ar18+, 146 eμA 86Kr28+, 0.5 eμA 86Kr33+, 53 eμA 129Xe38+, and 17 eμA 129Xe42+. Recent beam test results at SECRAL and SECRAL II have demonstrated that the production of more intense highly charged heavy ion beams needs higher microwave power and higher frequency, as the scaling law predicted. A 45 GHz superconducting ECR ion source FECR (a first fourth generation ECR ion source) is being built at IMP. FECR will be the world's first Nb3Sn superconducting-magnet-based ECR ion source with 6.5 T axial mirror field, 3.5 T sextupole field on the plasma chamber inner wall, and 20 kW at a 45 GHz microwave coupling system. This paper will focus on SECRAL performance studies at 24-28 GHz and technical design of 45 GHz FECR, which demonstrates a technical path for highly charged ion beam production from 24 to 28 GHz SECRAL to 45 GHz FECR.

Lawrence Berkeley National Laboratory in collaboration with the Institute of Modern Physics has developed a Nb3Sn-based superconducting magnet system for a fourth-generation electron cyclotron resonance source, with a goal of achieving the magnetic field required for operating at the microwave frequency of 45 GHz. The magnet system is composed of one sextupole magnet inside three solenoids of different sizes manufactured from Nb3Sn Sn round wire. Given the high stored energy density and relatively low wire copper fraction, the coils are not self-protected in the case of a quench. The study of the transient following a quench is carried out by means of the lumped-element dynamic electro-thermal program, which includes a detailed simulation of the interfilament coupling losses developing in the wire. Nonlinear effects occurring in the magnet, such as coupling loss and differential inductance reduction, have a significant impact on protecting these magnets. The resulting baseline quench protection strategy based on four independent energy extraction systems protecting the four magnets meets the quench protection requirements. Furthermore, in order to enhance the redundancy of the quench protection system and reduce the peak voltages to ground, the implementation of a coupling-loss induced quench (CLIQ) system is considered.

(1-x)Pb(Zn1/3Nb2/3)O3-xPbTiO3 ((1-x)PZN-xPT in short) is one of the most important piezoelectric materials. In this work, we extensively investigated (1-x)PZN-xPT (x = 0.07–0.11) ferroelectric single crystals using in-situ synchrotron μXRD, complemented by TEM and PFM, to correlate microstructures with phase transitions. The results reveal that (i) at 25 °C, the equilibrium state of (1-x)PZN-xPT is a metastable orthorhombic phase for x = 0.07 and 0.08, while it shows coexistence of orthorhombic and tetragonal phases for x = 0.09 and x = 0.11, with all ferroelectric phases accompanied by ferroelastic domains; (ii) upon heating, the phase transformation in x = 0.07 is Orthorhombic → Monoclinic → Tetragonal → Cubic. The coexistence of ferroelectric tetragonal and paraelectric cubic phases was in-situ observed in x = 0.08 above Curie temperature (TC), and (iii) phase transition can be explained by the evolution of the ferroelectric and ferroelastic domains. These results disclose that (1-x)PZN-xPT are in an unstable regime, which is possible factor for its anomalous dielectric response and high piezoelectric coefficient.

Haoming Chen,&ast; Jianmang Yu,&ast; Lihua Hang,&ast; Shuai Li, Weikang Lu, Zhenkai Xu Department of Anesthesiology, Kunshan First People’s Hospital Affiliated to Jiangsu University, Kunshan, Jiangsu, 215300, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Lihua HangDepartment of Anesthesiology, Kunshan First People’s Hospital Affiliated to Jiangsu University, Suzhou, Jiangsu Province, Peoples Republic of ChinaTel +86-18013273169Email zjhanglihua@foxmail.comIntroduction: Bone cancer pain (BCP) seriously affects the quality of life of patients with advanced cancer, but effective treatment methods are lacking. This study mainly investigates the role of EZH2 in a well-established BCP model induced by Walker 256 breast cancer cells in rats.Methods: Female Sprague–Dawley rats of the same age weighing approximately 160 g were selected for the experiment. The BCP model was established by injecting inactivated Walker 256 breast cancer cells into the tibia. von Frey filaments were used to measure the paw withdrawal threshold, and bone destruction in the rat was observed using x-ray. The spinal EZH2 and H3K27Tm levels were measured using Western blotting and RT–qPCR analysis. Intrathecal injection of an EZH2 inhibitor was performed to examine the role of EZH2 in trigeminal BCP.Results: Experimental results showed that injecting Walker 256 breast cancer cells into the tibia induced bone cancer pain. Spinal EZH2 and H3K27Tm levels were significantly increased over time in BCP rats. An intrathecal injection of 3-deazaneplanocin A (DZNep), a selective EZH2 inhibitor, downregulated the expression of EZH2 and attenuate the BCP-induced mechanical allodynia state.Conclusion: Intrathecal injection of DZNep relieve bone cancer pain in rats. EZH2 expressed in spinal cord tissue may be involved in the process of bone cancer pain in rats.Keywords: enhancer of zeste homologue 2, bone cancer pain, histone methyltransferase, 3 deazaneplanocin A, intrathecal injection

Lingwen Ying,1,&ast; Yong Zhang,2,&ast; Jun Yin,1,&ast; Yufei Wang,1 Wei Lu,1 Wei Zhu,1 Yuqian Bao,1 Jian Zhou1 1Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital; Shanghai Clinical Center for Diabetes; Shanghai Diabetes Institute; Shanghai Key Laboratory of Diabetes Mellitus, Shanghai, 200233, People’s Republic of China; 2Department of Immunology and Microbiology, Shanghai Jiao Tong University, School of Medicine, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jian ZhouDepartment of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, 600 Yishan Road, Shanghai, 200233, People’s Republic of ChinaTel +86-21-64369181Fax +86-21-64368031Email zhoujian@sjtu.edu.cnPurpose: We aimed to explore the immunological characteristics and cytokine profile of the initial stage of type 1 diabetes.Patients and Methods: In total, 123 age- and sex-matched subjects with newly diagnosed classic type 1 diabetes mellitus (T1ADM), fulminant type 1 diabetes mellitus (FT1DM), and normal glucose tolerance (NGT) were enrolled. Serum cytokine levels were measured using Milliplex MAP multifactor detection.Results: There was a significant increase in the levels of transforming growth factor β (TGFβ 1) and TGFβ 2 and decrease in programmed death-1 (PD-1), PD ligand 1 (PD-L1), pro-inflammatory cytokines, and anti-inflammatory cytokines in type 1 diabetes patients compared with the NGT subjects (all P < 0.05). There was no significant difference in C-reactive protein (CRP) and blood routine indicators between the two groups. Type 1 diabetes was further divided into T1ADM and FT1DM subgroups. FT1DM patients had much higher CRP levels than T1ADM patients (4.90 [0.95– 26.05] mg/L vs 0.39 [0.20– 0.74] mg/L, P < 0.01). Blood routine results showed that the number of leukocytes was significantly increased in FT1DM compared with that in T1ADM (9.2 [5.1– 18.8] × 109 cells/L vs 5.4 [4.5– 6.7] × 109 cells/L, P < 0.01). In FT1DM patients, neutrophil% was increased, and lymphocyte% was declined significantly, compared with that in T1ADM patients (neutrophil%: 80.2 [59.2– 85.2]% vs 59.5 [54.8– 64.0]%; lymphocyte%: 18.3 [10.1– 32.3]% vs 32.6 [26.8– 35.9]%; both P < 0.01). However, there was no difference between FT1DM and T1ADM in cytokine profile except for the decrease in CTLA-4 in T1ADM (P < 0.05).Conclusion: Compared with T1ADM, CRP and leukocytes’ levels were increased significantly in FT1DM, with an increase in neutrophil% and decline in lymphocyte%, suggesting that FT1DM may have more abrupt onset and occur as a more serious subtype of type 1 diabetes mellitus.Keywords: cytokine profile, fulminant type 1 diabetes mellitus, immune response, multifactor detection, type 1 diabetes mellitus

Hui Li,1,2,&ast; Xiao-Lei Fan,1,2,&ast; Yi-Nan Wang,1,2 Wei Lu,1,2 Haoyi Wang,1,2 Runzhi Liao,1,2 Min Zeng,1,2 Jun-Xiao Yang,1,2 Yihe Hu,1,2 Jie Xie1,2 1Department of Orthopedics, Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China; 2Hunan Engineering Research Center of Biomedical Metal and Ceramic Implants, Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jie Xie; Yihe HuDepartment of Orthopedics, Xiangya Hospital, Central South University, #87 Xiangya Road, Changsha, Hunan, 410008, People’s Republic of ChinaTel +86– 731– 89753006; +86– 731– 89753706Email dr_xiejie@163.com; csuhuyihe@163.comPurpose: Wear debris particle-induced periprosthetic osteolysis is a severe complication of total joint replacement that results in aseptic loosening and subsequent arthroplasty failure. No effective therapeutic agents or drugs have been approved to prevent or treat osteolysis; thus, revision surgery is often needed. Extracellular vesicles (EVs) are vital nanosized regulators of intercellular communication that can be directly applied to promote tissue repair and regeneration. In this study, we assessed the therapeutic potential of EVs from human urine-derived stem cells (USCs) (USC-EVs) in preventing ultrahigh-molecular-weight polyethylene (UHMWPE) particle-induced osteolysis.Methods: USCs were characterized by measuring induced multipotent differentiation and flow cytometry. USC-EVs were isolated and characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS) and Western blotting. RAW264.7 cells and bone marrow mesenchymal stem cells (BMSCs) were cultured with USC-EVs to verify osteoclast differentiation and osteoblast formation, respectively, in vitro. The effects of USC-EVs were investigated on a UHMWPE particle-induced murine calvarial osteolysis model by assessing bone mass, the inflammatory reaction, and osteoblast and osteoclast formation.Results: USCs differentiated into osteogenic, adipogenic and chondrogenic cells in vitro and were positive for CD44, CD73, CD29 and CD90 but negative for CD34 and CD45. USC-EVs exhibited a cup-like morphology with a double-layered membrane structure and were positive for CD63 and TSG101 and negative for calnexin. In vitro, USC-EVs promoted the osteogenic differentiation of BMSCs and reduced proinflammatory factor production and osteoclastic activity in RAW264.7 cells. In vivo, local injection of USC-EVs around the central sites of the calvaria decreased inflammatory cytokine generation and osteolysis compared with the control groups and significantly increased bone formation.Conclusion: Based on our findings, USC-EVs prevent UHMWPE particle-induced osteolysis by decreasing inflammation, suppressing bone resorption and promoting bone formation.Keywords: extracellular vesicles, urine-derived stem cells, UHMWPE, wear particle-induced osteolysis, anti-inflammatory

Min Xu,1,&ast; Dingchao Xiang,2,&ast; Wenhua Wang,1 Long Chen,1 Wei Lu,1 Feng Cheng3 1Department of Neurosurgery, Kunshan Hospital of Traditional Chinese Medicine, Kunshan Affiliated Hospital of Nanjing University of Chinese Medicine, Kunshan City, Jiangsu Province, 215300, People’s Republic of China; 2Department of Neurosurgery, Wuxi clinical medical school of Anhui Medical University, 904th Hospital of PLA(Taihu Hospital of Wuxi), Wuxi, 214000, People’s Republic of China; 3Department of Neurosurgery, Affiliated Kunshan Hospital of Jiangsu University, Kunshan, 215300, Jiangsu Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Feng ChengDepartment of Neurosurgery, Affiliated Kunshan Hospital of Jiangsu University, Kunshan, 215300, Jiangsu Province, People’s Republic of ChinaTel +86-13913208695Email fengchengjiangsu@163.comBackground: Nuclear factor erythroid 2-related factor 2 (Nrf2) is a key regulator responsible for oxidative stress in brain injury. This study aimed to investigate the potential mechanism of miR-448-3p and Nrf2 in cerebral ischemia/reperfusion (I/R) injury.Methods: In vitro and in vivo cerebral I/R injury models were constructed, and Nrf2 expression levels were detected by qRT-PCR and Western blot. The potential miRNAs for Nrf2 were predicted by bioinformatic analysis. The binding interaction between miR-448-3p and Nrf2 was determined by luciferase reporter assay. The effects of miR-448-3p on neurological deficit, infarct volume, and brain water content in mice were tested. The effects of miR-448-3p on oxidative stress indicators (SOD activity, MDA content, and ROS production) were detected by commercial assay kits. The levels of HO-1 and cleaved caspase-3 were evaluated by Western blot. Cell viability was evaluated by MTT assay, and cell apoptosis was evaluated by TUNEL staining and flow cytometry.Results: Nrf2 was significantly downregulated and miR-448-3p was upregulated in cerebral I/R injury both in vivo and in vitro. MiR-448-3p downregulation efficiently attenuated brain injury and reduced oxidative stress and apoptosis. MiR-448-3p was identified to act as ceRNA of Nrf2 and negatively regulated Nrf2 expression, which was consistent with the animal studies. In addition, Nrf2 silencing obviously attenuated the neuroprotective effects of miR-448-3p inhibitor in vitro.Conclusion: MiR-448-3p participated in the regulation of cerebral I/R injury via inhibiting Nrf2.Keywords: ischemia/reperfusion, miR-448-3p, Nrf2, oxidative stress, apoptosis