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Dandan Wang,1– 3,* Jingguo Wang,3,* Zequn Yin,4 Ke Gong,3 Shuang Zhang,3 Zhengbao Zha,3 Yajun Duan4 1School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, 230011, People’s Republic of China; 2State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University, Guiyang, 550014, People’s Republic of China; 3School of Food and Biological Engineering, Hefei University of Technology, Hefei, Anhui, 230601, People’s Republic of China; 4Department of Cardiology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230031, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhengbao Zha, School of Food and Biological Engineering, Hefei University of Technology, No. 420 Feicui Road, Hefei, Anhui, 230601, People’s Republic of China, Email zbzha@hfut.edu.cn Yajun Duan, Department of Cardiology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, No. 96 Jinzhai Road, Hefei, Anhui, 230031, People’s Republic of China, Email yajunduan@ustc.edu.cnIntroduction: Metabolic dysfunction-associated steatotic liver disease (MASLD), the most prevalent chronic liver disorder, has garnered increasing attention globally owing to its associated health complications. However, the lack of available therapeutic medications and inadequate management of complications in metabolic dysfunction-associated steatohepatitis (MASH) present significant challenges. There are little studies evaluating the effectiveness of POM in treating MASLD. In this study, we synthesized polyoxometalates (POM) for potential treatment of MASLD.Methods: We induced liver disease in mice using two approaches: feeding a high-fat diet (HFD) to establish MASLD or feeding a methionine–choline deficient (MCD) diet to induce hepatic lipotoxicity and MASH. Various metabolic parameters were detected, and biochemical and histological evaluations were conducted on MASLD. Western blotting, qRT-PCR and immunofluorescence assays were used to elucidate the molecular mechanism of POM in the treatment of MASLD.Results: POM therapy resulted in significant improvements in weight gain, dyslipidemia, liver injury, and hepatic steatosis in mice fed a HFD. Notably, in a more severe dietary-induced MASH model with MCD diet, POM significantly attenuated hepatic lipid accumulation, inflammation, and fibrosis. POM treatment effectively attenuated palmitic acid and oleic acid-induced lipid accumulation in HepG2 and Huh7 cells by targeting the AMPK pathway to regulate lipid metabolism, which was confirmed by AMPK inhibitor. Additionally, the activation of AMPK signaling by POM suppressed the expression of lipid synthesis genes, including sterol regulatory element-binding protein 1c (SREBP1c) and SREBP2, while concurrently upregulating the expression of sirtuin 1 (SIRT1) to promote fatty acid oxidation.Conclusion: These findings suggest that POM is a promising therapeutic strategy with high efficacy in multiple MASLD models. Keywords: polyoxometalates, MASLD, MASH, AMPK, lipotoxicity

Zhihong Zha,1,&ast; Shuang Gao,2,&ast; Shaofeng Liu,3 Guoqin Hu1 1Department of Otolaryngology Head and Neck Surgery, The Second People’s Hospital of Hefei, Hefei Hospital Affiliated to Anhui Medical University, Hefei, Anhui, People’s Republic of China; 2Anhui Public Health Clinical Center, The First Affiliated Hospital of Anhui Medical University North District, Hefei, Anhui, People’s Republic of China; 3Department of Otolaryngology Head and Neck Surgery, Yijishan Hospital of Wannan Medical College, Wuhu, Anhui, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Guoqin Hu, Department of Otolaryngology Head and Neck Surgery, The Second People’s Hospital of Hefei, Hefei Hospital Affiliated to Anhui Medical University, Hefei, Anhui, People’s Republic of China, Email huguoqin2010@yeah.netObjective: Chronic pharyngitis (CP) is a common disease, which has a long duration and a wide range of onset. Anxiety is considered to be a common complication in patients with CP. The purpose of this study was to evaluate the anxiety level and potentially influencing factors in patients with CP, to provide insights for the management of anxiety in patients with CP.Methods: A total of 104 adult patients with CP meeting the inclusion/exclusion criteria were enrolled from a single center in Wuhu, China between October 2015 and December 2016. A Self-rating Anxiety Scale (SAS) was used to measure the status of anxiety. The relationship between SAS scores and illness period in patients with CP was analyzed by Pearson correlation test. Univariate analysis and binary logistic regression analysis were performed to analyze the risk factors of anxiety in patients with CP.Results: The average SAS score of 104 patients with CP was (44.17 ± 8.38), including 82 cases (78.85%) without anxiety and 22 cases (21.15%) with anxiety. Moreover, the illness period was found to be positively related to SAS scores in patients with CP (r = 0.378, P = 0.001). Furthermore, the results of univariate analysis showed that there were significant differences in anxiety state among CP patients with different age, illness period, source of treatment payment, and marital status (P < 0.05). Additionally, the results of binary logistic regression analysis showed that age, source of treatment payment, and marital status were the independent risk factors affecting the anxiety status of patients with CP (P < 0.05).Conclusion: These results suggested that CP patients with advanced age, self-pay, and unmarried status had higher risk for anxiety.Keywords: chronic pharyngitis, risk factor, self-rating anxiety scale, anxiety, prevalence

Qiu Dong,1 Jialin Deng,2 Tsz Ngai Mok,1 Junyuan Chen,1 Zhengang Zha1 1Center for Bone, Joint and Sports Medicine, The First Hospital of Jinan University, Jinan University, Guangzhou, Guangdong, People’s Republic of China; 2School of Medicine, Jinan University, Guangzhou, Guangdong, People’s Republic of ChinaCorrespondence: Junyuan Chen; Zhengang ZhaCenter for Bone, Joint and Sports Medicine, The First Hospital of Jinan University, Jinan University, No. 613, Huangpu Avenue West, Guangzhou, 510515, Guangdong, People’s Republic of ChinaTel +86 134 8029 9854; +86 173 6182 0624Email chenjunyuan@jnu.edu.cn; zhazhengang1965@163.comBackground: Bone metastasis (BM) is the most common site of metastasis in non-small cell lung carcinoma (NSCLC). We aimed to construct and validate 2 novel nomograms predicting the 3-, 6-, and 12-months overall survival (OS) and cancer-specific survival (CSS).Methods: The clinical data of 7480 patients between 2010 and 2016 were enrolled from the Surveillance, Epidemiology, and End Results database (SEER). The patients were allocated randomly to training and validation cohorts in a 7:3 ratio. Cox proportional hazards regression models were used to identify prognostic risk factors and establish 2 nomograms. The prediction accuracy of nomograms was assessed by C-index, the area under the ROC curve (AUC), and calibration curves.Results: A total of 244998 NSCLC patients were identified between 2010 and 2016, with 7480 found with BM, accounting for 3.1%. Overall, 7480 patients were enrolled in the OS nomogram construction and were randomized to the training set (n = 5236) and the validation set (n = 2244). Age, sex, race, marital status, histology, grade, primary site, T stage, N stage, liver metastasis, surgery, radiotherapy, and chemotherapy were found to correlate with OS. A total of 7422 samples were included in the CSS nomogram construction, randomly grouped into training set (n = 5195) and the validation set (n = 2227). Age, sex, race, histology, grade, primary site, T stage, N stage, brain metastasis, liver metastasis, surgery, radiotherapy, and chemotherapy were associated with CSS. Two nomograms were conducted to predict the 3-, 6-, and 12-months OS and CSS. The ROC curves and exhibited good performance for predicting OS and CSS.Conclusion: We established and validated 2 high-performance nomograms to assist clinical doctors in making personalized treatment decisions.Keywords: lung cancer, metastasis, SEER, prognosis

Zhimin Zha,1,&ast; Yujia Cheng,2,3,&ast; Lu Cao,2 Yanxia Qian,2 Xinjian Liu,3,4 Yan Guo,1 Junhong Wang2 1Department of Gerontology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, People’s Republic of China; 2Department of Cardiology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, People’s Republic of China; 3Key Laboratory of Antibody Techniques of National Health Commission, Nanjing Medical University, Nanjing, People’s Republic of China; 4Department of Pathogen Biology, Nanjing Medical University, Nanjing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yan Guo; Junhong WangDepartment of Gerontology, The First Affiliated Hospital with Nanjing Medical University, No. 300 Guangzhou Road, Gulou District, Nanjing City, Jiangsu Province, 210009, People’s Republic of ChinaTel +86-25-68305083; +86-25-68308273Email guoyan51@hotmail.com; wangjunhong@jsph.org.cnObjective: A polarized macrophage response plays a critical role in the pathophysiological process of myocardial infarction (MI). Several studies have shown a pro-inflammatory role for monomeric C-reactive protein (mCRP) in cardiovascular disease. However, the mechanism of how mCRP regulates macrophage phenotype switching remains unknown. In the present study, the effect of mCRP on macrophage polarization and its pathological function in myocardial repair after myocardial infarction was investigated.Methods: MI was induced by permanent ligation of the left anterior descending coronary artery in ICR mice. Adult mice were injected with mCRP (2.5 mg/kg) with or without SP600125 (15 mg/kg, JNK inhibitor) 45 min before MI. The cardiac function, scar size as well as cardiac fibrosis, infiltration of inflammatory cells, and the level of proteins in the JNK signaling pathway in infarcted myocardium were assessed. In addition, the phenotypic characterization of macrophages was further measured by ELISA, flow cytometry and quantitative RT-PCR in cultured THP-1 cells or peritoneal macrophages.Results: Cardiac function deterioration, ventricular dilatation and fibrosis were exacerbated in mice pretreatment with mCRP following MI. Meanwhile, an increased accumulation of infiltrated inflammatory cells in infarcted myocardium was observed in the mCRP group. Moreover, activation of the JNK signaling pathway was markedly elevated in mCRP treated animals post-MI. In contrast, pharmacological inhibition of JNK phosphorylation activity by SP600125 muted the detrimental effects of mCRP in MI mice. Furthermore, in vitro and in vivo co-culture experiments showed that mCRP shifted macrophage polarization towards pro-inflammatory phenotypes, and this polarization could be abolished by sp600125.Conclusion: Taken together, our results imply that mCRP impairs myocardial repair after myocardial infarction by polarizing the macrophages into the pro-inflammatory M1 phenotype via the JNK-dependent pathway.Keywords: myocardial infarction, monomeric CRP, inflammation, macrophage polarization, JNK pathway

Zhaozhen Zhang, Zhuqing Zha, Zhiwei Zhao, Wenjing Liu, Wuyin Li Department of Bone Surgery, Luoyang Orthopedic Hospital of Henan Province (Henan Provincial Orthopedic Hospital), Zhengzhou City, Henan Province 450000, People’s Republic of ChinaCorrespondence: Wuyin LiDepartment of Bone Surgery, Luoyang Orthopedic Hospital of Henan Province (Henan Provincial Orthopedic Hospital), No. 100, Yongping Road, Zhengzhou City, Henan Province 450000, People’s Republic of ChinaTel +86-371-88820000Email wuyinl128@163.comBackground: Chondrocyte-mediated inflammation is an important pathological component of osteoarthritis (OA) development. There are currently no therapies that completely reverse the development of OA. Lentinan, a type of polysaccharide derived from Lentinus edodes, has been demonstrated to possess significant anti-viral, anti-cancer, and anti-inflammatory effects, and has been recently used in the treatment of several inflammatory diseases. However, little research has focused on the pharmacological effect of lentinan in human OA.Materials and Methods: We evaluated the anti-inflammatory and anti-ROS effects of lentinan in SW1353 chondrocytes treated with AGEs using real-time polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and the nitro oxide-specific stain DAF-FM DA. The regulatory effects of lentinan on NF-κB and MAPK p38 signaling were investigated via promoter assay and Western blot analysis.Results: We found that lentinan inhibits the production of pro-inflammatory cytokines, including IL-1β, TNF-α, IL-8 and the secretion of PGE2 and NO, by reducing the expression of COX-2 and iNOS in AGE-challenged chondrocytes. Lentinan also reduces AGE-induced increased expression of matrix metalloproteinases-1, − 3, and − 13 (MMP-1, MMP-3, MMP-13). Furthermore, lentinan has a similar effect on a disintegrin and metalloproteinase with thrombospondin motifs-4 and − 5 (ADAMTS-4, ADAMTS-5). Mechanistically, lentinan reduces the activation of NF-κB.Conclusion: Our findings indicate that lentinan shows a protective effect against AGE-induced inflammatory response in chondrocytes. These findings suggest that lentinan is a promising agent for the treatment of OA that could be used as a dietary supplement for patients with OA.Keywords: osteoarthritis, AGEs, lentinan, NF-κB, chondrocytes

Songwei Huan,1,* Tao Gui,1,* Qiutong Xu,1,* Songkuan Zhuang,2 Zhenyan Li,1 Yuling Shi,3 Jiebin Lin,3 Bin Gong,1 Guiqiang Miao,1 Manseng Tam,4 Huan-Tian Zhang,1 Zhengang Zha,1 Chunfei Wu3 1Institute of Orthopedic Diseases and Department of Bone and Joint Surgery, The First Affiliated Hospital, Jinan University, Guangzhou 510630, Guangdong, People’s Republic of China; 2School of Life Science, Xiamen University, Xiamen, Fujian 361005, People’s Republic of China; 3Department of Orthopedics, The Third Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510405, People’s Republic of China; 4IAN WO Medical Center, Macao Special Administrative Region, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhengang Zha; Chunfei Wu Tel +86-20-38688617; Tel +86-13610075186Fax +86-20-38688000; Email tbri@jnu.edu.cn; wuchunfei666@163.comBackground: Chondrosarcoma is the second-most common type of bone tumor and has inherent resistance to conventional chemotherapy. Present study aimed to explore the therapeutic effect and specific mechanism(s) of combination BET family protein and HDAC inhibition in chondrosarcoma.Methods: Two chondrosarcoma cells were treated with BET family protein inhibitor (JQ1) and histone deacetylase inhibitors (HDACIs) (vorinostat/SAHA or panobinostat/PANO) separately or in combination; then, the cell viability was determined by Cell Counting Kit-8 (CCK-8) assay, and the combination index (CI) was calculated by the Chou method; cell proliferation was evaluated by 5-ethynyl-2′-deoxyuridine (EdU) incorporation and colony formation assay; cell apoptosis and reactive oxygen species (ROS) level were determined by flow cytometry; protein expressions of caspase-3, Bcl-XL, Bcl-2, γ-H2AX, and RAD51 were examined by Immunoblotting; DNA damage was determined by comet assay; RAD51 and γ-H2AX foci were observed by immunofluorescence.Results: Combined treatment with JQ1 and SAHA or PANO synergistically suppressed the growth and colony formation ability of the chondrosarcoma cells. Combined BET and HDAC inhibition also significantly elevated the ROS level, followed by the activation of cleaved-caspase-3, and the downregulation of Bcl-2 and Bcl-XL. Mechanistically, combination treatment with JQ1 and SAHA caused numerous DNA double-strand breaks (DSBs), as evidenced by the comet assay. The increase in γ-H2AX expression and foci formation also consistently indicated the accumulation of DNA damage upon cotreatment with JQ1 and SAHA. Furthermore, RAD51, a key protein of homologous recombination (HR) DNA repair, was found to be profoundly suppressed. In contrast, ectopic expression of RAD51 partially rescued SW 1353 cell apoptosis by inhibiting the expression of cleaved-caspase-3.Conclusion: Taken together, our results disclose that BET and HDAC inhibition synergistically inhibit cell growth and induce cell apoptosis through a mechanism that involves the suppression of RAD51-related HR DNA repair in chondrosarcoma cells.Keywords: chondrosarcoma, JQ1, HDAC, apoptosis, RAD51, DNA repair

Lijin Zhang, Bin Wu, Zhenlei Zha,* Hu Zhao,* Yejun Feng* Department of Urology, Affiliated Jiang-Yin Hospital of the Southeast University Medical College, Jiang-Yin 214400, China *These authors contributed equally to this work Background and purpose: Numerous studies have demonstrated that sarcomatoid differentiation is linked to the risk of renal cell carcinoma (RCC). However, its actual clinicopathological impact remains inconclusive. Therefore, we undertook a meta-analysis to evaluate the pathologic and prognostic impacts of sarcomatoid differentiation in patients with RCC by assessing cancer-specific survival, overall survival, recurrence-free survival, progression-free survival, and cancer-specific mortality. Materials and methods: In accordance with the preferred reporting items for systematic reviews and meta-analysis statement, relevant studies were collected systematically from PubMed, Embase, and Web of Science to identify relevant studies published prior to January 2018. The pooled effects (hazard ratios, odds ratios, and standard mean differences) and 95% confidence intervals were calculated to investigate the association of sarcomatoid differentiation with cancer prognosis and clinicopathological features. Results: Thirty-five studies (N=11,261 patients [n=59–1,437 per study]) on RCC were included in this meta-analysis. Overall, the pooled analysis suggested that sarcomatoid differentiation was significantly associated with unfavorable cancer-specific survival (HR=1.46, 95% CI: 1.26–1.70, p

KDM2B (also known as FBXL10) controls stem cell self-renewal, somatic cell reprogramming and senescence, and tumorigenesis. KDM2B contains multiple functional domains, including a JmjC domain that catalyzes H3K36 demethylation and a CxxC zinc-finger that recognizes CpG islands and recruits the polycomb repressive complex 1. Here, we report that KDM2B, via its F-box domain, functions as a subunit of the CUL1-RING ubiquitin ligase (CRL1/SCF(KDM2B)) complex. KDM2B targets c-Fos for polyubiquitylation and regulates c-Fos protein levels. Unlike the phosphorylation of other SCF (SKP1-CUL1-F-box)/CRL1 substrates that promotes substrates binding to F-box, epidermal growth factor (EGF)-induced c-Fos S374 phosphorylation dissociates c-Fos from KDM2B and stabilizes c-Fos protein. Non-phosphorylatable and phosphomimetic mutations at S374 result in c-Fos protein which cannot be induced by EGF or accumulates constitutively and lead to decreased or increased cell proliferation, respectively. Multiple tumor-derived KDM2B mutations impaired the function of KDM2B to target c-Fos degradation and to suppress cell proliferation. These results reveal a novel function of KDM2B in the negative regulation of cell proliferation by assembling an E3 ligase to targeting c-Fos protein degradation that is antagonized by mitogenic stimulations.

Because of their convenience, rapidness and non-invasive advantages, non-invasive skin physiological monitoring technique can be auxiliary examination of many common skin diseases and is of great significance in guiding the diagnosis and treatment of diseases. Skin physiological monitor is a newly-developed non-invasive skin physiological monitoring technique, which can evaluate skin barrier by measuring transepidermal water loss rate (TEWL), skin surface pH, stratum corneum hydration (SCH), sebum content and so on. This manuscript reviews and discusses the applications and significance of non-invasive skin physiological monitors in dermatology. It will provide a basis for analyzing the correlation between skin barrier function and diagnostic and therapeutic outcomes.

[Objective] Achieving non-destructive testing of sub-healthy watercore apples. [Methods] First, the logarithmic function method and the power function method proposed by this study were used to correct the sample spectra. Subsequently, the corrected data were converted into different images of gramian angular difference field (GADF), gramian angular summation field (GASF), markov transition field (MTF), recurrence plot (RP), symmetric dot pattern (SDP). Then, the ResNet50 network model with the convolutional block attention module (CBAM) was used to mine the deep image features related to the degree of watercore, which were downscaled by the t-distributed stochastic neighbor embedding (t-SNE) method and analyzed by clustering to determine the most suitable image transformation method. Finally, the most suitable image features were inputted into the improved particle swarm algorithm (IPSO) optimized support vector machine (SVM), extreme learning machine (ELM), k-nearest neighbour (KNN) and random forest (RF) classifier for the three-class classification of watercore apple. [Results] The results showed that the power function method was better than the logarithmic function method in eliminating the effect of diameter on the transmission spectrum. The silhouette coefficient (SC), the calinski harabasz score (CHS), and the davies-bouldin index (DBI) were 0.93, 0.88 and 0.24 after visualization of the image features in the GADF, better than the rest of the image transformation methods. ResNet50-IPSO-ELM achieved the highest classification accuracy of 96.8%. The overall discrimination accuracy of the three watercore classes apples in the test set reached 96.3%, and the stable precision (SP), stable recall (SR), and stable F1-score (SF) were 87.2%, 95.8%, and 92.3%, respectively. [Conclusion] The model maintains a high classification accuracy for the majority class of apples without watercore and healthy apples with watercore and a high discriminatory ability for the minority class of sub-healthy apples with watercore.

OBJECTIVE: To investigate whether microRNA-588 was involved in the development and progression of renal cancer, and to explore its possible regulatory mechanisms. PATIENTS AND METHODS: Tumor tissues excised from renal carcinoma and adjacent normal tissues were selected for the experiment. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to analyze the expression level of microRNA-588 in tissue specimens. The relationship between the expression of microRNA-588 and the prognosis of patients with renal cell carcinoma was also evaluated. Subsequently, two renal cancer cell lines, including769-P and 786-O, were selected for functional experiments in vitro. Eukaryotic initiation factor 5A2 (pcDNA-EIF5A2) or microRNA-588 mimics was transfected into 769-P cells, respectively. Meanwhile, si-EIF5A2 or microRNA-588 inhibitor was transfected into 786-O cells. After that, the mRNA expression level of EIF5A2 was detected by qRT-PCR. The invasiveness and metastasis abilities of the two cell lines were evaluated via transwell assay. Furthermore, the levels of EIF5A2 and epithelial- mesenchymal transition (EMT)-related proteins were analyzed using Western blot. Luciferase reporter gene assay was used to confirm that microRNA-588 could directly regulate EIF5A2 expression. QRT-PCR and Western blot were performed to explore the mRNA and protein expressions of EIF5A2 in patients with highly or lowly-expressed microRNA-588. The correlation between the two molecules was evaluated using linear analysis. Through the above experiments, it was verified whether microRNA-588 could enhance the invasiveness and metastasis of renal cancer by targeting EIF5A2. RESULTS: MicroRNA-588 expression in tumor tissues of patients with renal carcinoma was significantly decreased with the increase of tumor diameter and stage. A higher level of microRNA-588 indicated significantly longer overall survival of patients. This suggested that microRNA-588 expression was negatively correlated with the prognosis of patients. Overexpression of microRNA-588 remarkably reduced the invasion and metastasis abilities of 769-P cells, as well as the expressions of EMT-related proteins. However, opposite results were observed in 786-O cells after knockdown of microRNA- 588. Reporter gene assay confirmed that microRNA-588 could target bind to EIF5A2. In 769-P cells, up-regulated microRNA-588 significantly inhibited the mRNA and protein expressions of EIF5A2. However, down-regulated microRNA-588 in 786-O cells significantly enhanced the expressions of EIF5A2 at both mRNA and protein levels. Linear analysis verified that microRNA-588 was negatively correlated with EIF5A2 at the mRNA level. Additionally, the up-regulation of EIF5A2 in 769-P cells enhanced the malignancy of cancer cells and the expressions of EMT-related proteins. However, in 786- O cells, opposite results were observed after knockdown of EIF5A2. CONCLUSIONS: MicroRNA-588 was lowly expressed in renal cancer tissues and cell lines. This might lead to an increase in the protein level of EIF5A2, eventually promoting tumor invasion and metastasis. [ABSTRACT FROM AUTHOR]

Wang Tang1 &ast;, Zhen-Wei Li1 &ast;, Gui-Qiang Miao2 &ast;, Zhi-Peng Li,1 Tao Gui,1 Chong-Jie Wu,1 Zhen-Yan Li,1 Jie Yang,1 Xiao-Dong Zhao,2 Ning Liu,1 Zhen-Gang Zha,1 Lu-Tian Yao,3 Huan-Tian Zhang1 1Department of Bone and Joint Surgery, The First Affiliated Hospital of Jinan University, Guangzhou, 510630, People’s Republic of China; 2Department of Orthopedics, Foshan Fosun Chancheng Hospital, Foshan, 528010, People’s Republic of China; 3Department of Orthopedics, The First Hospital of China Medical University, Shenyang, 110001, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Lu-Tian Yao; Huan-Tian Zhang, Email ltyao@cmu.edu.cn; zhanghuantian@jnu.edu.cnPurpose: Subchondral insufficiency fracture of the knee (SIFK) is a common cause of knee joint pain that mainly afflicts the elderly. Until now, how a sudden insufficiency fracture of subchondral bone affects the transcriptomic profiles of cartilage in SIFK and OA patients are largely unknown.Methods: Single-cell RNA sequencing (scRNA-seq) was used to identify various cell subsets and evaluate transcriptomic differences in cartilage of SIFK and OA patients. In addition, the above findings were confirmed by histological evaluation and immunohistochemical (IHC) staining.Results: We found that the transcriptomic profiles of cartilage in the SIFK patient was completely different from those of normal and OA patients. Accordingly, several novel cell clusters with activation of hypoxia and endochondral ossification signaling were identified in the SIFK cartilage. Chondrocyte trajectories analysis and IHC staining revealed that transcription factors including TCF4 were found to be highly up-regulated during the occurrence of SIFK, which might drive the reactive formation of cartilage and fibrous tissue and the activation of endochondral ossification.Conclusion: This is the first report to elucidate the transcriptomic alterations and distinct cell type subpopulations in the cartilage of SIFK and OA by the use of scRNA-seq, which provides a new insight in the understanding of the initiation and progression of SIFK.Keywords: SIFK, scRNA, TCF4, cartilage, hypoxia

OBJECTIVE: To explore the influences of micro ribonucleic acid (miR)-34a on liver function and hepatocyte proliferation during hepatocyte regeneration in rats and its mechanism. MATERIALS AND METHODS: A total of 80 Sprague-Dawley rats were randomly divided into 4 groups: Sham-2 d group (2 days after hepatectomy), Sham-10 d group (10 days after hepatectomy), miR-34a siRNA-2d group (miR- 34a knockdown + 2 days after hepatectomy) and miR-34a siRNA-10 d group (miR-34a knockdown + 10 days after hepatectomy), with 20 rats in each group. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were detected at 2 d and 10 d after the operation. The rat liver was harvested for calculating the liver/body weight ratio. In addition, the deoxyribonucleic acid (DNA) content in rat hepatocytes was detected via Feulgen staining. The pathological changes in rat liver were detected via hematoxylin- eosin (H&E) staining. Moreover, the hepatocyte apoptosis in each group was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Expression levels of proliferating cell nuclear antigen (PCNA), Notch1 intracellular domain (NICD), and hypoxia-inducible factor-1α (HIF-1α) in liver tissues of each group were detected via immunohistochemistry and Western blotting. RESULTS: No significant differences in the liver/body weight ratio, serum levels of ALT, AST, LDH, pathological structure of the liver, hepatocyte apoptosis level, and PCNA expression in hepatocytes were found between miR-34a siRNA-2 d group and Sham-2 d group. However, the expression levels of NICD and HIF-1α in the liver significantly increased in miR-34a siRNA- 2 d group compared with those in Sham-2 d group (p<0.05). On the contrary, compared with those in Sham-10 d group, the liver function and hepatocyte regeneration level significantly increased in miR-34a siRNA-10 d group. Increased liver/body weight ratio, remarkable decline in serum levels of ALT, AST, and LDH, significant alleviation of pathological injury of liver tissues, decreased the apoptosis level and upregulated PCNA protein were observed in miR-34a siRNA- 10 d group than those of Sham-10 d group. The Notch/HIF-1α signaling pathway was also significantly activated. CONCLUSIONS: MiR-34a knockdown can significantly enhance the liver function and hepatocyte regeneration ability in rats at 10 d after hepatectomy through activating the Notch/HIF-1α signaling pathway. [ABSTRACT FROM AUTHOR]

The capability to support large wave vector bulk plasmon polariton (BPP) waves enables the application of hyperbolic metamaterials (HMMs) in sensing. However, there is a challenge arising from the excitation of BPP, and the highly confined polarization waves are unable to meet the requirements of practical application. In this study, an HMM/bilayer silver nanoparticles (Ag NPs) platform is proposed that allows the excitation and utilization of BPP for use as a surface-enhanced Raman scattering (SERS) substrate. According to the research results, the bilayer Ag NPs provide stronger plasmonic property and act as a light-matter coupler, so as to generate a large wave vector of scattered light and excite the BPP within the HMM. Besides, Ag NPs provide the nano antenna structure, and decouple the BPP into localized surface plasmon (LSP) that can be used directly to excite the electric fields. In addition, HMM produces a modulating effect on the plasmon resonance peak, which makes it possible to overlap the spectrum of resonance peak with excitation wavelengths, thus leading to a strong absorption peak at the incident laser wavelength region. Experimentally, the platform was applied to achieve SERS detection for adenosine molecules with a concentration of 10−6 M. It is believed that this plasmonic platform has a potential of application in surface-enhanced spectroscopy.