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Zhongyuan Li,1,&ast; Pengfei Xu,2,&ast; Yang Deng,3,&ast; Rui Duan,1 Qiang Peng,1 Shiyao Wang,1 Zhaohan Xu,1 Ye Hong,1 Yingdong Zhang1 1Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210000, People’s Republic of China; 2Department of Neurology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, People’s Republic of China; 3Department of Neurology, Nanjing First Hospital, China Pharmaceutical University, Nanjing, 210006, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ye Hong; Yingdong Zhang, Department of Neurology, Nanjing First Hospital, Nanjing Medical University, No. 68, Changle Road, Nanjing, 210000, People’s Republic of China, Email dg1735053@smail.nju.edu.cn; zhangyingdong@aliyun.comBackground: After ischemic stroke (IS), microglia and astrocytes undergo polarization, transforming into a pro-inflammatory phenotype (M1 or A1). According to previous studies, exosomes might play an important role in the interplay between M1 microglia and A1 astrocytes after IS.Methods: We used the microglial oxygen-glucose deprivation/reperfusion (OGD/R) model and ultracentrifugation to extract M1 microglial exosomes (M1-exos). Subsequently, we identified circSTRN3 enriched in exosomes through RNA sequencing and detected the role of circSTRN3 in astrocyte activation based on bioinformatics analysis, immunofluorescence, Western blotting, and polymerase chain reaction analysis. We validated these findings in the middle cerebral artery occlusion/reperfusion (MCAO/R) model of adult male C57BL/6J mice. Finally, we confirmed the correlation among circSTRN3, miR-331-5p, and stroke severity score in exosomes isolated from peripheral blood of IS patients.Results: Our findings revealed that M1-exos promoted A1 astrocyte activation. CircSTRN3 was abundant in M1-exos, which could sponge miR-331-5p to affect mitochondrial antiviral signaling protein (MAVS), activate NF-κB pathway, and participate in A1 astrocyte activation. In addition, overexpressed circSTRN3 augmented the infarct size and neurological dysfunction in MCAO/R models, while miR-331-5p mimics reversed the effect. Furthermore, circSTRN3 in IS patients was positively correlated with stroke severity score (R2 = 0.83, P < 0.001), while miR-331-5p demonstrated a negative correlation with the same score (R2 = 0.81, P < 0.001).Conclusion: Taken together, our research indicated that circSTRN3 from M1-exos could promote A1 astrocyte activation and exacerbate ischemic brain injury via miR331-5p/MAVS/NF-κB axis.Keywords: ischemic stroke, microglia, astrocyte, exosome, circSTRN3, miR-331-5p

Ruidong Ma,&ast; Wei Li,&ast; Qiufeng Peng,&ast; Ao Ren, Ling Zhao, Jiawei Li, Shiqiao Luo Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Shiqiao Luo, Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Youyi Road 1st, Yuzhong District, Chongqing, 400016, People’s Republic of China, Tel +86-15002355050, Email 202065@hospital.cqmu.edu.cnPurpose: Prior research has suggested a correlation between gallstones and depressive symptoms, yet the specifics of this relationship remain unclear. This study aims to explore the association between gallstones and depressive symptoms among adults.Patients and Methods: Initially, we conducted a cross-sectional study using data from the National Health and Nutrition Examination Survey (NHANES) 2017 – March 2020. After propensity score matching (PSM) for participants with gallstones and those without gallstones, multivariate logistic regression analysis was used to explore the potential association between gallstones and depressive symptoms. This was followed by Mendelian randomization (MR) analysis to further elucidate the causal relationship between them. Using the genome-wide association study database, we extracted instrumental variables and performed bidirectional univariate and multivariate MR analyses.Results: In the cross-sectional study of NHANES 2017 – March 2020, 835 pairs of participants with comparable characteristics, both with and without gallstones, were identified after PSM. The multivariate adjusted logistic regression analyses revealed a significant association between gallstones and depressive symptoms [fully adjusted model: OR=1.821 (95% CI, 1.181– 2.808), P=0.007]. Subsequent MR analyses further clarified the causal relationship, indicating that genetically determined gallstones significantly increase the risk of developing depressive symptoms [forward univariate MR analysis: OR=1.04 (95% CI, 1.01– 1.06), P=0.002; multivariate MR analysis: OR=1.03 (95% CI, 1.01– 1.05), P=0.009], with no evidence of reverse causation [inverse univariate MR analysis: OR=1.28 (95% CI, 0.90– 1.83), P=0.17].Conclusion: Gallstones are a risk factor for depressive symptoms among adults. Hence, we recommend timely depression screening for patients diagnosed with gallstones, facilitating early detection and effective treatment of depressive symptoms, thus alleviating its impact on both individuals and society.Keywords: depressive symptoms, gallstones, risk factor, causality, genetic analysis, cross-sectional study

Yang Deng,1,&ast; Rui Duan,2,&ast; Ye Hong,2,&ast; Qiang Peng,2 Zhong-Yuan Li,2 Xiang-Liang Chen,2 Ying-Dong Zhang1 1Department of Neurology, Nanjing First Hospital, China Pharmaceutical University, Nanjing, 210006, People’s Republic of China; 2Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiang-Liang Chen, Department of Neurology, Nanjing First Hospital, Nanjing Medical University, No. 68, Changle Road, Nanjing, Jiangsu, 210006, People’s Republic of China, Email chenxl@njmu.edu.cn Ying-Dong Zhang, Department of Neurology, Nanjing First Hospital, China Pharmaceutical University, No.68, Changle Road, Nanjing, Jiangsu, 210006, People’s Republic of China, Email zhangyingdong@njmu.edu.cnPurpose: Vascular dementia (VaD) is the second most common dementia in the world. An increasing number of studies have demonstrated the important role of long non-coding RNAs (lncRNAs) in VaD. Our previous investigation demonstrated that Trimethylamine-N-oxide (TMAO) exacerbates cognitive impairment and neuropathological alterations in VaD rats. Thus, we hypothesized that TMAO could play an injury role in VaD by regulating lncRNAs.Materials and Methods: The rats using the bilateral common carotid artery (2VO) model were administered TMAO (120 mg/kg) for 8 consecutive weeks, 4 weeks preoperatively and 4 weeks postoperatively. High-throughput sequencing was conducted to investigate the effects of TMAO treatment on lncRNA expression in rat hippocampus and bioinformatics analysis was performed to identify potential downstream targets. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect the levels of lncRNA fetal-lethal noncoding developmental regulatory RNA (Fendrr), miR-145-5p, and paxillin (PXN). Learning and spatial memory capacities were measured, as well as inflammatory factors. Nissl staining was used to observe neuronal injury in the CA1 area of the hippocampus. Furthermore, we used the Fendrr loss-of-function assay, miR-145-5p gain-of-function assays and PXN loss-of-function assay to explore the mechanisms by which TMAO acts on VaD.Results: TMAO administration upregulated lncRNA Fendrr expression in the rat hippocampus, while the damaging effects of TMAO were counteracted after knockdown of Fendrr. Fendrr exhibits highly expressed in 2VO rats and sponged miR-145-5p, which targets PXN. Silencing of Fendrr or PXN, or promotion of miR-145-5p improved neurological function injury, reduced neuronal damage, as well as repressed inflammation response. Inhibition of miR-145-5p abrogated up Fendrr knockdown mediated influence on 2VO rats.Conclusion: The results of this study indicated that TMAO inhibits the miR-145-5p/PXN axis by increasing the Fendrr expression, thus exacerbating the development of VaD.Keywords: trimethylamine N-oxide, neuroinflammation, Fendrr, MiR-145-5p, PXN, vascular dementia

Cai-Dong Liu,1,2,&ast; Qiang Peng,3,&ast; Shi-Yao Wang,3,&ast; Yang Deng,2 Zhong-Yuan Li,3 Zhao-Han Xu,3 Liang Wu,3 Ying-Dong Zhang,2,3 Rui Duan3 1Department of Laboratory Medicine, Nanjing First Hospital, China Pharmaceutical University, Nanjing, Jiangsu, 210006, People’s Republic of China; 2School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, 210006, People’s Republic of China; 3Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, 210006, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ying-Dong Zhang; Rui Duan, Department of Neurology, Nanjing First Hospital, Nanjing Medical University, No. 68, Changle Road, Nanjing, Jiangsu, People’s Republic of China, Email zhangyingdong@njmu.edu.cn; duanruicpu@163.comPurpose: Acute ischemic stroke (AIS) has seriously threatened people’s health worldwide and there is an urge need for early diagnosis and effective treatment of AIS. This research intended to clarify the regulatory role of circ_0008146/miR-342-5p/ACSL4 axis in AIS.Methods: High-throughput small RNA sequencing analysis was adapted to identify differentially expressed miRNAs between the AIS and control group. The circ_0008146, miR-342-5p, and ACSL4 levels were detected by qRT-PCR. Middle cerebral artery occlusion/reperfusion (MCAO/R) models were constructed in C57BL/6J mice. Assay kits were used to determine Fe2+ levels and a battery of oxidative stress and lipid peroxidation indicators, including ROS, MDA, LPO, SOD and GSH/GSSG ratio. The protein levels of ACSL4 were measured by Western blot. The behavioral function was assessed using neurobehavioral tests. TTC staining was employed to visualize infarction size. Nissl staining was adapted to detect histopathological changes. Receiver operating characteristic curve and correlation analysis were applied to investigate the clinical value and association of miR-342-5p and ACSL4.Results: A total of 44 AIS patients and 49 healthy controls were enrolled in our study. The small RNA sequencing unveiled a significant decrease in miR-342-5p levels in AIS patients. MiR-342-5p inhibited oxidative stress and RSL3-induced ferroptosis after cerebral ischemic/reperfusion injury in vivo by targeting ferroptosis-related gene ACSL4. Circ_0008146 acted as a sponge of miR-342-5p, and overexpression of circ_0008146 increased neurological deficits and brain injury in mice. Circ_0008146 contributed to ferroptosis in cerebral infarction via sponging miR-342-5p to regulate ACSL4. Plasma miR-342-5p and ACSL4 demonstrated significant correlation and good diagnostic value for AIS patients.Conclusion: This study provides the first in vivo evidence to show that circ_0008146 exacerbates neuronal ferroptosis after AIS via the miR-342-5p/ACSL4 axis. Furthermore, miR-342-5p/ACSL4 axis holds promise as a viable therapeutic target and practical biomarkers for AIS patients.Keywords: ischemic stroke, miRNA, circRNA, ferroptosis, biomarker

Jiaxing Zhang,1,2,&ast; Liwei Gu,3,&ast; Yumao Jiang,4,&ast; Yun Ma,5,&ast; Ziyue Zhang,3 Shengnan Shen,3 Shuo Shen,3 Qing Peng,6 Wei Xiao2,7 1Institute of Digestive Disease, The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People’s Hospital, Qingyuan, Guang Dong, People’s Republic of China; 2School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guang Dong, People’s Republic of China; 3Artemisinin Research Center and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China; 4Gannan Medical University, Ganzhou, Jiang Xi, People’s Republic of China; 5Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou, Guang Dong, People’s Republic of China; 6Institute of Basic Medical Sciences, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China; 7Key Laboratory of Glucolipid Metabolic Disorder, Ministry of Education, Guangdong Pharmaceutical University, Guangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Qing Peng; Wei Xiao, Email pengqing85@163.com; xw7688@smu.edu.cnBackground: Acute kidney injury (AKI) is a syndrome, posing a substantial healthcare burden. The pathological basis of AKI is associated with inflammation and oxidative stress which cause additional damage to mitochondria. Artesunate (ATS) is a derivative of artemisinin isolated from Artemisia annua L. that is an effective treatment for malaria and favored for the prevention and treatment of kidney diseases. However, there are still challenges related to its efficacy, including poor water solubility, limited oral bioavailability and short half-life. Liposome-based nanoparticles are used for drug delivery due to their ideal biocompatibility and their ability to improve the bioavailability of specific drugs and enhance drug efficacy.Methods: In this study, a novel TPP-based natural ATS-nanoliposome, namely T-A-Ls, was applied for the treatment of AKI. ATS was encapsulated with or without triphenylphosphonium (TPP)-modified nanoliposomes. AKI was induced by cisplatin in C57BL/6J mice and a cisplatin-induced injury model was generated in HK-2 cells in vitro. Blood urea nitrogen (BUN), serum creatinine (Scr) measurements and section staining were utilized to assess renal protective effect of T-A-Ls. Inflammatory-related factors and proteins were quantified via Elisa, Immunofluorescence and Western Blot (WB). The anti-mitochondrial oxidative stress effect of T-A-Ls was determined by ROS, JC-1 and oxygen consumption rate (OCR) kits. Immunohistochemistry and WB were conducted to measure associated protein expressions. In vivo biodistribution and the concentration of T-A-Ls in kidney were also explored.Results: T-A-Ls exhibited good oxidative resistance, preferential renal uptake, mitochondrial targeting, and it ameliorated kidney injury in cisplatin-induced AKI mice. Mitochondrial dysfunction, ATP production and respiratory capacity were improved in damaged HK-2 cells; ROS content decreased while mitochondrial membrane potential recovered. T-A-Ls exerted renal protection by inhibiting inflammation and reducing oxidative stress; these effects were mediated by a downregulation in the expression of RAGE and iNOS and an upregulation in both Nrf2 and HO-1.Conclusion: T-A-Ls could improve the delivery of ATS to the kidney, offering a promising avenue to treat AKI. Keywords: artesunate, liposomes, AKI, cisplatin, oxidative stress, inflammation

Chengyu Yang,1,2,&ast; Li Yang,3,4,&ast; Lei Yang,5 Shuiming Li,5 Ling Ye,1,6 Jinfeng Ye,5 Chengshui Chen,3,4,7 Yiming Zeng,8 Mengchan Zhu,1 Xiaoping Lin,8 Qing Peng,9 Yun Wang,5,10 Meiling Jin1,6 1Department of Pulmonary and Critical Care Medicine, Zhongshan Hospital, Fudan University, Shanghai, 200032, People’s Republic of China; 2Department of Pulmonary and Critical Care Medicine, Huadong Hospital, Fudan University, Shanghai, 200040, People’s Republic of China; 3Department of Pulmonary and Critical Care Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015, People’s Republic of China; 4Key Laboratory of Interventional Pulmonology of Zhejiang Province, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015, People’s Republic of China; 5Longhua Innovation Institute for Biotechnology, Shenzhen University, Shenzhen, Guangdong, 518055, People’s Republic of China; 6Department of Allergy, Zhongshan Hospital, Fudan University, Shanghai, 200032, People’s Republic of China; 7Department of Pulmonary and Critical Care Medicine, the Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People’s Hospital, Zhejiang, 324000, People’s Republic of China; 8Department of Pulmonary and Critical Care Medicine, the Second Affiliated Hospital of Fujian Medical University, Respiratory Medicine Center of Fujian Province, Quanzhou, Fujian, 362000, People’s Republic of China; 9Department of Pulmonary and Critical Care Medicine, Minhang Hospital, Fudan University, Shanghai, 201199, People’s Republic of China; 10Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, Shantou, Guangdong, 515041, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Meiling Jin, Department of Pulmonary and Critical Care Medicine, Zhongshan Hospital, Fudan University, Shanghai, 200032, People’s Republic of China, Email jin.meiling@zs-hospital.sh.cn Yun Wang, Longhua Innovation Institute for Biotechnology, Shenzhen University, Shenzhen, Guangdong, 518055, People’s Republic of China, Email yunw@szu.edu.cnBackground: Frequent exacerbation (FE) and infrequent exacerbation (IE) are two phenotypes of chronic obstructive pulmonary disease (COPD), of which FE is associated with a higher incidence of exacerbation and a serious threat to human health. Because the pathogenesis mechanisms of FE are unclear, this study aims to identify FE-related proteins in the plasma via proteomics for use as predictive, diagnostic, and therapeutic biomarkers of COPD.Methods: A cross-sectional study was conducted in which plasma protein profiles were analyzed in COPD patients at stable stage, and differentially expressed proteins (DEPs) were screened out between the FE and IE patients. FE-related DEPs were identified using data-independent acquisition-based proteomics and bioinformatics analyses. In addition, FE-related candidates were verified by enzyme-linked immunosorbent assay.Results: In this study, 47 DEPs were screened out between the FE and IE groups, including 20 upregulated and 27 downregulated proteins. Key biological functions (eg, neutrophil degranulation, extracellular exosome, protein homodimerization activity) and signaling pathways (eg, arginine and proline metabolism) were enriched in association with the FE phenotype. Receiver operating characteristic (ROC) analysis of the 11 combined DEPs revealed an area under the curve of 0.985 (p < 0.05) for discriminating FE from IE. Moreover, correlation and ROC curve analyses indicated that creatine kinase, M-type (CKM) and fat storage-inducing transmembrane protein 1 (FITM1) might be clinically significant in patients with the FE phenotype. In addition, plasma expression levels of CKM and FITM1 were validated to be significantly decreased in the FE group compared with the IE group (CKM: p < 0.01; FITM1: p < 0.05).Conclusion: In this study, novel insights into COPD pathogenesis were provided by investigating and comparing plasma protein profiles between the FE and IE patients. CKM, FITM1, and a combinative biomarker panel may serve as useful tools for assisting in the precision diagnosis and effective treatment of the FE phenotype of COPD.Keywords: chronic obstructive pulmonary disease, frequent exacerbation, data-independent acquisition, proteomics, bioinformatics analysis

Chunli Fan,1,2,&ast; Quanfu Ma,1,2,&ast; Xufeng Wu,2 Xuan Dai,2 Qiuzi Peng,2 Hongning Cai1,2 1Department of Scientific Research and Teaching Management Department, Maternal and Child Health Hospital of Hubei Province, Wuhan, 430070, People’s Republic of China; 2Department of Cervical Cancer Prevention Center, Maternal and Child Health Hospital of Hubei Province, Wuhan, 430070, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Qiuzi Peng; Hongning Cai, Email 1078380199@qq.com; chn@hbfy.comObjective: To evaluate the diagnostic value of DNA methylation detection of multiple gene loci in cervical cancer.Methods: A total of 61 cases requiring cervical biopsy were selected from the outpatient clinic of Maternal and Child Health Hospital of Hubei Province between January 2018 and December 2019. The patients were divided into four groups based on histopathologic diagnosis: cervical cancer (CC) group, high-grade squamous intraepithelial lesion (HSIL) group, low-grade squamous intraepithelial lesion (LSIL) group, and control group. HPV examination, liquid-based cytology examination, and DNA methylation detection at multiple gene sites were performed. The positive rate of DNA methylation, sensitivity, specificity, area under the curve (AUC), and other efficacy indexes were calculated to evaluate the diagnostic value of DNA methylation detection at multiple gene loci in cervical cancer.Results: The positive rates of DNA methylation in CC, HSIL, LSIL, and control groups were 100%, 88%, 83% and 17%, respectively. The ZNF671 gene had the highest positive rate among the cervical lesion group, with rates of 57%, 76%, and 100% in LSIL, HSIL, and CC groups respectively. The combination of DNA methylation detection at multiple gene loci showed the highest diagnostic efficacy for HSIL and cervical cancer, with AUC value of 0.850 (95% CI:0.746– 0.954), a Youden index of 0.654, and a sensitivity and specificity of 85% and 85.4%, respectively. The diagnostic efficacy of the combined detection was significantly higher than that of HPV examination and liquid-based cytology examination (P < 0.05).Conclusion: DNA methylation detection at multiple gene loci is highly effective and diagnostic tool for cervical cancer, and has potential application value in clinical practice.Keywords: multiple gene loci, DNA methylation detection, HSIL and cervical cancer, diagnostic efficacy

Wanyang Dong,1,&ast; Qi Peng,1,&ast; Xiaohua Tang,1,2,&ast; Tian Zhong,1 Shunan Lin,1 Ziling Zhi,1 Jingyi Ye,1 Bixia Yang,1 Ning Sun,1,3 Wenchang Yuan1 1Guangzhou Key Laboratory for Clinical Rapid Diagnosis and Early Warning of Infectious Diseases, KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 510180, People’s Republic of China; 2Third Affiliated Hospital of Guangzhou Medical University, Department of Clinical Laboratory, Guangzhou, 510150, People’s Republic of China; 3Guangzhou First People’s Hospital, School of Medicine, South China University of Technology, Guangzhou, 510180, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ning Sun; Wenchang Yuan, Email ning.sun@connect.polyu.hk; yuanwenchang95@163.comBackground: Staphylococcus haemolyticus is an opportunistic pathogen that belongs to coagulase-negative Staphylococci (CoNS). Increasing infection and multi-drug resistance cases caused by this strain have been reported and thus it poses a great health threat.Methods: The third-generation sequencing technology was performed on a S. haemolyticus SH-1 isolated from a clinical sample to analyze the drug resistance genes, which included vancomycin resistance related genes. In addition, antimicrobial susceptibility tests, transmission electron microscopy and Triton X-100 stimulated autolysis were conducted to understand its biological characteristics.Results: The study shows that this clinical isolate is a vancomycin intermediate-resistant strain. Genome comparison also revealed that WalK(N70K) and WalK(R280Q) mutations may contribute to the vancomycin resistant phenotype. Besides, S. haemolyticus SH-1 exhibit common features of thicker cell wall and decreased autolytic activity.Conclusion: S. haemolyticus SH-1 with WalKR mutations shows typical characteristics of vancomycin resistant strains. Combining the genome features and biological properties, our findings may provide important information for the understanding of the molecular mechanism of S. haemolyticus to vancomycin intermediate-resistance.Keywords: Staphylococcus haemolyticus, drug resistance gene, vancomycin intermediate-resistance, whole genome sequencing

Yin-Gen Luo, Xiao-Wu Zhang, Jiay-Wei Tsauo, Ying Li, Jin-Gui Li, Qing Peng, Ya-Wei Li, Wei Sun, He Zhao, Xiao Li Department of Interventional Therapy, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People’s Republic of ChinaCorrespondence: Xiao Li; He Zhao, Department of Interventional Therapy, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, People’s Republic of China, Tel/Fax +86 010 8778 8502, Email simonlixiao@gmail.com; hezhaoo@gmail.comObjective: This study aimed to investigate the shortest compression time to achieve hemostasis and the optimal hemostasis strategy in patients treated with transradial access chemoembolization (TRA-TACE).Methods: From October 2019 to October 2021, 119 consecutive patients with hepatocellular carcinoma (HCC) who underwent 134 sessions of TRA-TACE were included in this prospective single-center study. The compression time was measured by decompressing the device for 30 min, and thereafter, every 10 min after the procedure until complete hemostasis was achieved.Results: Technical success was achieved for all TRA procedures. None of the patients experienced major TRA-related adverse events. Minor adverse events occurred in 7.5% of the patients. The mean compression time was 31.8 ± 5.0 min. Factors that may impact hemostasis were analyzed by univariate and multivariate analyses, and a platelet count < 100× 109 /L (p = 0.016, odds ratio = 3.942) was found to be an independent factor that could predict the failure to achieve hemostasis within 30 min. For patients with a platelet count < 100× 109 /L, the compression time required to achieve hemostasis was 60 min. For patients with a platelet count ≥ 100× 109 /L, the compression time required to achieve hemostasis was 40 min.Conclusion: To achieve hemostasis in patients with HCC treated with TRA-TACE, compression for 60 min is sufficient for those with a platelet count < 100× 109 /L, and compression for 40 min is sufficient for those with a platelet count ≥ 100× 109 /L.Keywords: hepatocellular carcinoma, transarterial chemoembolization, transradial access, hemostasis

XiaoJin Sun,1,&ast; Yang Deng,2,&ast; PengXin Ge,3,&ast; Qiang Peng,4 Ismatullah Soufiany,4 Lin Zhu,4 Rui Duan4 1Faculty of Pharmacy, Bengbu Medical College, Anhui Engineering Technology Research Center of Biochemical Pharmaceuticals, Bengbu, Anhui, People’s Republic of China; 2School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, People’s Republic of China; 3Department of Pharmacy, Anhui Provincial Cancer Hospital, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, People’s Republic of China; 4Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Lin Zhu; Rui Duan, Email julinzz@163.com; duanruicpu@163.comPurpose: ACE2/Ang(1– 7)/Mas Receptor, the momentous component of the renin-angiotensin system, has been shown to be involved in Alzheimer’s disease (AD). We had previously found that enhancing brain ACE2 activity ameliorated cognitive impairment and attenuated brain neuroinflammation in SAMP8 mice, an animal model of AD. However, the exact mechanism of action of Diminazene (DIZE) has not been revealed.Methods: APP/PS1 mice were injected intraperitoneally with DIZE. Cognitive functions, neuronal and synaptic integrity, and inflammation-related markers were assessed by Morris water maze, Nissl staining, Western blotting and ELISA, respectively. Since astrocytes played a crucial role in AD-related neuroinflammation whilst miRNAs were reported to participate in modulating inflammatory responses, astrocytes of APP/PS1 mice were then isolated for high-throughput miRNAs sequencing to identify the most differentially expressed miRNA following DIZE treatment. Afterward, the downstream pathway of this miRNA in the anti-inflammatory action of DIZE was investigated using primary astrocytes.Results: The results showed that DIZE alleviated cognitive impairment and neuronal and synaptic damage in APP/PS1 mice. Simultaneously, DIZE suppressed the secretion of pro-inflammatory cytokines and the expression of NLRP3 inflammasome. Importantly, miR-224-5p was significantly up-regulated in the astrocytes of APP/PS1 mice treated by DIZE, and NLRP3 is one of the targets of miR-224-5p. Upregulation of miR-224-5p inhibited the expression of NLRP3 in Aβ 1– 42-stimulated cells, whereas miR-224-5p downregulation reversed this effect. Furthermore, the inhibition of miR-224-5p could reverse the inhibitory effect of DIZE on astrocytic NLRP3 inflammasome.Conclusion: These findings firstly suggested that DIZE could inhibit astrocyte-regulated neuroinflammation via miRNA-224-5p/NLRP3 pathway. Furthermore, our study reveals the underlying mechanism by which DIZE suppresses neuroinflammatory responses in AD mice and uncovers the potential of DIZE in AD treatment.Keywords: DIZE, miRNA-224-5p, astrocyte, NLRP3, neuroinflammation, Alzheimer’s disease

Qiang Peng,1,&ast; Wenxiu Chen,2,&ast; Yan E,1,&ast; Yang Deng,3 Zhaohan Xu,1 Siyu Wang,1 Xinxin Fu,3 Bin Wei,1 Meng Wang,1 Jiankang Hou,1 Yingdong Zhang,1 Rui Duan1 1Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, 210006, People’s Republic of China; 2Department of Critical Care Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, 210006, People’s Republic of China; 3School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, 210000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yingdong Zhang; Rui Duan, Department of Neurology, Nanjing First Hospital, Nanjing Medical University, No. 68 Changle Road, Nanjing, 210006, People’s Republic of China, Email zhangyingdong@njmu.edu.cn; duanruicpu@163.comPurpose: Neuron-specific enolase (NSE) is considered a biomarker for the severity of nervous system diseases. We sought to explore whether serum NSE concentration in ischemic stroke patients undergoing mechanical thrombectomy (MT) is related to 3-month functional outcome and symptomatic intracranial hemorrhage (sICH).Patients and Methods: We retrospectively collected the data of acute ischemic stroke patients with anterior circulation infarction receiving MT within 6 h in our stroke center. Favorable outcome and poor outcome at 3 months were defined as modified Rankin Scale (mRS) score 0– 2 and 3– 6, respectively. sICH was defined according to the Heidelberg bleeding classification. We used multivariate logistic regression model and receiver operating characteristic curves to investigate the correlation between NSE and clinical outcomes.Results: Among the 426 patients enrolled, 40 (9.4%) patients developed sICH. Three-month favorable outcome in 160 (37.6%) and poor outcome in 266 (62.4%) patients were observed. Serum NSE levels was significantly correlated with 3-month mRS score (R = 0.473, P < 0.001). A cutoff value of 15.29 and 23.12 ng/mL for serum NSE was detected in discriminating 3-month poor outcome (area under the curve, 0.724) and sICH (area under the curve, 0.716), respectively. Multivariate analysis showed that high serum NSE levels were independently associated with 3-month poor outcome (odds ratio [OR] 5.049, 95% confidence interval [CI] 2.933– 8.689, P< 0.001) and sICH (OR 5.111, 95% CI 2.210– 11.820, P < 0.001).Conclusion: Our study demonstrated that high serum NSE levels after receiving MT were independently associated with 3-month poor outcome and sICH in acute ischemic stroke patients. Serum NSE levels could be a good predictor of clinical outcomes for patients receiving MT.Keywords: neuron-specific enolase, ischemic stroke, mechanical thrombectomy, symptomatic intracranial hemorrhage, prognosis

Yang Deng,1,&ast; Si-Yu Wang,2,&ast; Qing-Guang Wang,3,&ast; Zhao-Han Xu,2 Qiang Peng,2 Shuai-Yu Chen,2 Lin Zhu,2 Ying-Dong Zhang,1,2 Rui Duan1,2 1Department of Neurology, Nanjing First Hospital, China Pharmaceutical University, Nanjing, People’s Republic of China; 2Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, People’s Republic of China; 3Department of Neurology, Jiangyin Hospital Affiliated to Nantong University, Jiangyin, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ying-Dong Zhang; Rui Duan, Department of Neurology, Nanjing First Hospital, China Pharmaceutical University, No.68, Changle Road, Nanjing, Jiangsu, People’s Republic of China, Email zhangyingdong@njmu.edu.cn; duanruicpu@163.comPurpose: Our previous study has shown that AVE 0991, a nonpeptide analogue of Ang-(1-7), ameliorates cognitive decline and inhibits NLRP3 inflammasome of astrocytes in Alzheimer’s disease model mice. Additionally, several studies have suggested that activation of autophagy appears to effectively inhibit the progression of neuroinflammation. However, it is unclear whether AVE 0991 can modulate astrocyte autophagy to suppress neuroinflammation in Alzheimer’s disease.Materials and Methods: APP/PS1 mice and Aβ-treated primary astrocytes were used as the research objects in vivo and in vitro, respectively. Water maze test was used to evaluate cognitive function of mice, Nissl staining and immunofluorescence staining was used to assess neuronal damage. ELISA kits were used to detect the levels of Ang-(1-7) and Aβ in the cortex, and qRT-PCR was used to detect the expression of cortical inflammation-related mediators. The expression of autophagy-related proteins in cortex were detected by Western blot. The upstream molecular responses involved in inflammation inhibition by AVE 0991 were validated by means of using the Mas1 antagonist and autophagy inhibitor.Results: We found that 30 days of intraperitoneal administration of AVE 0991 improved. Aβ deposition, neuronal death, and cognitive deficits in APP/PS1 Alzheimer’s disease model mice. Moreover, AVE 0991 treatment greatly suppressed astrocyte-mediated inflammation and up-regulated the expression of autophagy. Furthermore, the inhibitory effect of AVE 0991 on the expression of inflammatory factors was reversed by 3-MA, an autophagy inhibitor.Conclusion: These findings suggest that regulation of autophagy is critical for inhibiting astrocyte neuroinflammatory responses and demonstrate a potential neuroprotective mechanism by which AVE 0991 could suppress neuroinflammatory responses by enhancing autophagy.Keywords: AVE 0991, neuroinflammation, Alzheimer, autophagy, astrocytes

Objective To explore the application of deep-learning technology in automatic classification of intraventricular hemorrhage. Methods The brain CT images of 1 027 patients with spontaneous ICH from the First Affiliated Hospital of Army Medical University from January 2010 to December 2020 were collected and retrospectively analyzed, which were subsequently divided into 3 types: normal, intracerebral hemorrhage with/without intraventricular extension.Six typical deep networks, including DenseNet121, ResNet101, ResNet50, Swin-base, Vit-base and VGG16, were used to construct classification models for identifying whether there was intraventricular extension of intracerebral hemorrhage.The performance of each model was assessed on both the internal and external data sets (CQ500) respectively.In order to enhance the interpretability of the deep learning networks, the EigenGradCAM method was adopted to generate heatmaps for visualization of the interested regions of the model. Results The performance of the models was evaluated based on precision, recall rate, specificity, negative predictive value (NPV) and F1 value.On the internal test set of VGG16 model, the normal group achieved 0.983, 0.977, 0.984, 0.978 and 0.980, respectively; the intracerebral hemorrhage without intraventricular extension group achieved 0.917, 0.902, 0.965, 0.958 and 0.909;and the intraventricular extension of intracerebral hemorrhage group obtained 0.877, 0.911, 0.966, 0.976 and 0.894, respectively.Meanwhile, on the external test set of VGG16 model, the normal group reached 0.967, 0.870, 0.985, 0.938 and 0.916;the intracerebral hemorrhage without intraventricular extension group reached 0.827, 0.939, 0.902, 0.967 and 0.879;and the intraventricular extension of intracerebral hemorrhage group achieved 0.938, 0.906, 0.970, 0.954 and 0.922.The accuracy (ACC) of internal and external test sets was 0.940 and 0.905, respectively.Finally, the heatmaps generated by EigenGradCAM method showed the VGG16 could reasonably focus on the relevant areas. Conclusion The VGG16 achieved the best predictive performance for identifying whether there is an intraventricular extension in intracerebral hemorrhage or not, indicating that deep learning can be effectively applied to the judgement of intraventricular hemorrhage.

Zhendong Li,1,&ast; Chenliang Zhou,2,&ast; Qing Peng,3,&ast; Suguo Wang,3 Guowei Qian,2 Lina Tang,2 Xin Zhou,4 Qingcheng Yang,1 Zan Shen,2 GaoZhong Huang,3 Yonggang Wang,2 Hongtao Li2 1Department of Orthopedics, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai, People’s Republic of China; 2Department of Oncology, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai, People’s Republic of China; 3Department of VIP Clinic, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai, People’s Republic of China; 4Cancer Research Institute of Jilin University, the First Hospital of Jilin University, Changchun, Jilin, 130021, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yonggang Wang; Hongtao Li, Department of Oncology, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, No. 600, Yishan Road, Xu Hui District, Shanghai, 200233, People’s Republic of China, Tel +086-021-24058408, Email sirui666@163.com; Lhtmed@126.comPurpose: Inflammatory response and nutritional status are associated with cancer development and progression. The present study aimed to evaluate the predictive ability of the fibrinogen–albumin ratio index (FARI) to the efficacy of neoadjuvant chemotherapy (NAC) for osteosarcoma.Patients and methods: A retrospective analysis involving 752 consecutive osteosarcoma patients between 2012 and 2020 was performed. Data on serum fibrinogen, albumin levels, white blood cell count, platelet count, and alkaline phosphatase (ALP) before and after NAC were collected. The predictive value of the NAC efficacy in osteosarcoma was assessed by constructing a receiver operating characteristic (ROC) curve and calculating the area under the curve (AUC). Prognosis and its predictive factors were analyzed by Kaplan–Meier method and COX regression analysis. Nomogram was established according to selected variables. The predictive performance of the nomogram model was assessed using C-statistics.Results: A total of 203 patients were included. ROC analysis showed that both FARI before NAC (preFARI; AUC = 0.594, p = 0.032) and the change in FARI before and after NAC (dfFARI = preFARI-postFARI; AUC = 0.652, p = 0.001) exhibited more favorable predictive ability than ALP and other inflammation markers. The preFARI was divided into the high group (> 6.1%) and the low group (≤ 6.1%) based on the optimal cut-off value of 6.1%. Patients with a high preFARI showed significantly decreased metastasis-free survival (MFS) and disease-free survival (DFS) (all p< 0.01). In multivariable analysis, preFARI was an independent prognostic marker for patients with osteosarcoma. Predictive nomograms exhibited good ability to predict MFS (C-index = 0.748, se = 0.028) and DFS (C-index=0.727, se = 0.030).Conclusion: Our findings indicated that FARI exhibits the favorable predictive ability for the efficacy of NAC for osteosarcoma, which could support clinicians and patients in clinical decision-making and treatment optimization.Keywords: osteosarcoma, neoadjuvant chemotherapy, fibrinogen, albumin, fibrinogen–albumin ratio index, prognosis

Tao Zhang,1,&ast; Qin Liu,2,&ast; Yingfan Zhu,1 Songfa Zhang,3 Qiaohua Peng,3 Amanda Louise Strickland,4 Wenxin Zheng,4,5 Feng Zhou2 1Department of Gynecology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of China; 2Department of Pathology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of China; 3Department of Oncology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of China; 4Department of Pathology, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA; 5Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX, USA&ast;These authors contributed equally to this workCorrespondence: Feng ZhouDepartment of Pathology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou City, Zhejiang Province, 310006, People’s Republic of ChinaTel +86-571-89991702Fax +86-571-87061878Email fungchew@zju.edu.cnPurpose: Programmed death-ligand 1 (PD-L1) has been widely used as a prognostic biomarker and an immunotherapeutic target in numerous cancers, but information on the clinical significance of its expression in endometrial serous carcinoma (ESC) is largely lacking. Here, we evaluate the predictive value of PD-L1 expression in ESC.Materials and Methods: A total of 79 cases of ESC accessioned between January 2003 and September 2015 were selected for further analysis. PD-L1 expression was evaluated in whole tissue sections of these cases by using the tumor proportion score (TPS, cut-off 1%) and combined positive score (CPS, cut-off 1) scoring methods.Results: Overall, there was a heterogeneous expression of PD-L1, focal or patchy, in ESCs. PD-L1 positivity was observed in 43.0% of ESCs by TPS and 73.4% of ESCs by CPS. Kaplan–Meier survival analysis showed that patients with PD-L1-positive tumors suffered significantly worse OS and PFS, when compared with PD-L1 negative tumors (log-rank p = 0.037 and p = 0.003, respectively). In contrast, PD-L1 positivity by CPS within the ESC cases showed no statistical significance for OS and PFS (log-rank p = 0.720 and p = 0.928, respectively). Multivariate Cox analysis showed that PD-L1 positivity by TPS was significantly associated with PFS (HR = 1.921, p = 0.039) but not OS (HR = 1.229, p = 0.631).Conclusion: PD-L1 expression is frequently found in ESC, suggesting a potential role of the PD-1/PD-L1 pathway as a potential therapeutic target for these tumors. PD-L1 expression by TPS also serves as a negative prognostic marker in ESC and implies an unfavorable outcome.Keywords: PD-L1, endometrial serous carcinoma, endometrial cancer, immune therapy

Ruhai Bai,1 Wanyue Dong,2 Jinli Liu,3 Qiao Peng,3 Jun Lyu4 1School of Public Affairs, Nanjing University of Science and Technology, Nanjing, People’s Republic of China; 2School of Health Economics and Management, Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China; 3Health Science Center, Xi’an Jiaotong University, Xi’an, People’s Republic of China; 4Department of Clinical Research, The First Affiliated Hospital of Jinan University, Guangzhou, People’s Republic of ChinaCorrespondence: Jun LyuDepartment of Clinical Research, The First Affiliated Hospital of Jinan University, No. 601, West Huangpu Avenue, Guangzhou, Guangdong, People’s Republic of ChinaEmail lyujun2020@jnu.edu.cnPurpose: The Belt and Road Initiative (BRI) provided a platform to promote trade and economic collaboration, with health promotion also being an important component. This study examined the burden of the under-5 mortality rate (U5MR) among BRI countries and studied the impact of socioeconomic development on the U5MR.Patients and Methods: Data were collected from 137 BRI countries for the period 2000– 2018. The temporal changes in the U5MR and the estimated annual percentage change (EAPC) were used to assess the U5MR burden. A quantile fixed-effects model was used to access the associations between socioeconomic factors and the U5MR.Results: The U5MR varied widely among the 137 analyzed countries in 2018, being highest in Somalia (121.5 per 1000) and lowest in Slovenia (2.1 per 1000). The decreasing trend in U5MR was largest in Montenegro (EAPC = – 9.6, 95% confidence interval [CI] = – 10.2 to – 9.0), while the increasing trend was largest in Dominica (EAPC = 6.0, 95% CI = 5.3 to 6.7). Countries with higher U5MRs in 2000 experienced more rapid decreases in the U5MR from 2000 to 2018. The influence of socioeconomic factors on U5MR was related to the distribution of U5MR.Conclusion: The U5MR remains a major public-health issue in some BRI countries. Improving the economic situation will benefit child health in BRI countries in the long run.Keywords: Belt and Road, under-five mortality rate, socioeconomic, economic factors

Mi Zhang,1 Wei Zhao,2 Shanshan Liu,1 Haichuan Liu,1 Liang Liu,2 Qiling Peng,2 Chengyou Du,1 Ning Jiang3 1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China; 2School of Basic Medical Science, Chongqing Medical University, Chongqing, 400016, People’s Republic of China; 3Department of Pathology, Chongqing Medical University, Chongqing, 400016, People’s Republic of ChinaCorrespondence: Chengyou Du; Ning Jiang Email duchengyou@126.com; jiangning@cqmu.edu.cnBackground: The H/ACA small nucleolar ribonucleoprotein (snoRNP) gene family, including GAR1 ribonucleoprotein (GAR1), NHP2 ribonucleoprotein (NHP2), NOP10 ribonucleoprotein (NOP10), and dyskerin pseudouridine synthase 1 (DKC1), play important roles in ribosome biogenesis. However, the potential clinical value of the H/ACA snoRNP gene family in hepatocellular carcinoma (HCC) has not yet been reported.Methods: Bioinformation databases were used to analyze the expression and roles of the H/ACA snoRNP gene family in HCC. Survival analysis, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes enrichment pathway (KEGG) analyses were performed using R software. Tumor Immune Estimation Resource (TIMER) was used to analyze the correlation between the expression of the H/ACA snoRNP gene family and immune infiltration in HCC. Finally, immunohistochemistry and Western blotting were performed to verify the protein expression of the H/ACA snoRNP gene family in HCC tissues and adjacent tissues.Results: The expression of the H/ACA snoRNP gene family was significantly increased in HCC samples compared to normal tissues, and the area under the curve (AUC) of GAR1, NHP2, NOP10, and DKC1 was 0.898, 0.962, 0.884, and 0.911, respectively. Increased expression of the H/ACA snoRNP gene family was associated with poor prognosis in HCC patients (Hazard Ratio, HR = 1.44 [1.02– 2.04], 1.70 [1.20– 2.40], 1.53 [1.09– 2.17], and 1.43 [1.02– 2.03], respectively; log-rank P = 0.036, 0.003, 0.014, 0.039, respectively). GO and KEGG analyses showed that co-expressed genes were primarily enriched in ribosome biogenesis. In addition, upregulated expression of H/ACA snoRNP gene family was related to the infiltration of various immune cells and multiple T cell exhaustion markers in HCC patients. Immunohistochemical analysis and Western blotting showed that the protein expression of H/ACA snoRNP gene family was higher in HCC tissues than in adjacent tissues of clinical samples.Conclusion: H/ACA snoRNP gene family expression was higher in HCC tissues than in normal or adjacent tissues and was highly associated with poor prognosis of HCC patients and, therefore, has the potential to serve as diagnostic and prognostic biomarkers for HCC.Keywords: hepatocellular carcinoma, H/ACA snoRNP gene family, diagnosis, prognosis, biomarker

Tingting Sun, 1,* Qingquan Wei, 1,* Peng Gao, 1,* Yongjie Zhang, 2 Qing Peng 1 1Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University, Shanghai, 200072, People’s Republic of China; 2Department of Human Anatomy, Nanjing Medical University, Nanjing, Jiangsu Province, 211166, People’s Republic of China*These authors contributed equally to this workCorrespondence: Qing PengDepartment of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University, Shanghai, 200072, People’s Republic of ChinaEmail pengqing@tongji.edu.cnYongjie ZhangDepartment of Human Anatomy, Nanjing Medical University, Nanjing, Jiangsu, 211166, People’s Republic of ChinaEmail zhangyongjie@njmu.edu.cnObjective: To investigate the concentrations of cytokine and chemokines profiling in aqueous humor for choroidal neovascularization (CNV) due to neovascular age-related macular degeneration (nAMD) before and during Intravitreal injection of ranibizumab (IVR) and its relation with the disease’s active state.Methods: The cytokine levels in aqueous humour were detected by the Bio-Plex® 200 System and the Bio-Plex™ Human Cytokine Standard 27-Plex, Group I. Aqueous humour samples of experimental group were collected from 19 patients diagnosed nAMD at baseline and at 1 month after IVR. Aqueous humour samples of control group were collected from 20 patients undergoing cataract surgery.Results: Aqueous humor levels of basic fibroblast growth factor (basic FGF) and RANTES were significantly lower in nAMD patients than in the control group (P=0.044 and P< 0.001, respectively). Vascular endothelial growth factor-A (VEGF-A) was significantly higher in nAMD patients than in the control group (P < 0.001). The average Eotaxin levels were significantly higher in nAMD patients after IVR than before (P=0.03). Contrarily, the average VEGF-A levels were significantly lower in AMD patients after IVR than before (P < 0.001).Conclusion: Angiogenic, growth factors and inflammatory are involved in the formation of neovascularization of AMD patients. IVR did not cause significant differences in any growth factors or inflammatory cytokines in nAMD patients with the exception of VEGF.Keywords: choroidal neovascularization, CNV, neovascular age-related macular degeneration, nAMD, ranibizumab, cytokines

Ting Zhang,1 Yanping Li,2 Yi Song,1 Xiaoshuang Chen,1 Jing Li,1 Qiang Peng,3 Jinhan He,1,2 Xiaofan Fei1 1Department of Pharmacy, West China Hospital, Sichuan University, Chengdu 610041, People’s Republic of China; 2Laboratory of Clinical Pharmacy and Adverse Drug Reaction, West China Hospital of Sichuan University, Chengdu, People’s Republic of China; 3State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, People’s Republic of ChinaCorrespondence: Xiaofan Fei Tel + 86-28-85423475Email 1145342867@qq.comBackground and Purpose: Hepatic fibrosis is a public health problem characterized by activation of hepatic stellate cells (HSCs), which triggers excessive production of extracellular matrix (ECM). Inhibition of HSC activation may be an effective treatment. Since various pathways control HSC activation, a combination of drugs with different mechanisms may be more effective than monotherapy.Methods: Here, we prepared liposomes loaded with curcumin and cyclopamine to inhibit HSC activation. We systematically analyzed the physicochemical characteristics of liposomes loaded with the two drugs, as well as their effects on HSC proliferation, activation and collagen production on gene, protein and cellular levels.Results: The prepared liposomes helped solubilize both drugs, contributing to their uptake by cells. Liposomes loaded with both drugs inhibited cell proliferation, migration and invasion, as well as induced more apoptosis and perturbed the cell cycle more than the free combination of both drugs in solution or liposomes loaded with either drug alone. Liposomes loaded with both drugs strongly suppressed HSC activation and collagen secretion.Conclusion: Our results suggest that liposome encapsulation can increase the uptake of curcumin and cyclopamine as well as the synergism between them in anti-fibrosis. This approach shows potential for treating hepatic fibrosis.Keywords: hepatic stellate cells, HSCs, hepatic fibrosis, cyclopamine, curcumin, synergistic effect

Huimin Li,1,* Qian Peng,2,* Yisha Guo,3 Xiaohui Wang,1 Li Zhang1 1Department of Pharmacy, Logistics College of Chinese People’s Armed Police Forces, Tianjin 300309, People’s Republic of China; 2Jiangsu Hengrui Pharmaceutical Co. LTD, Jiangsu 222000, People’s Republic of China; 3Characteristic Medical Center of the Chinese People’s Armed Police Forces, Tianjin 300162, People’s Republic of China*These authors contributed equally to this workCorrespondence: Li Zhang; Xiaohui WangDepartment of Pharmacy, Logistics College of Chinese People’s Armed Police Forces, Tianjin 300309, People’s Republic of ChinaTel +86-13302086775; +86-13302023963Email zhli62tianjin@163.com; 1517183319@qq.comPurpose: Asiaticoside (ASI), a compound of triterpene pentacyclic saponins, has apparently therapeutic efficacy on human hypertrophic scar. However, the characteristics of large molecular weight, low water solubility and poor lipophilicity do not favor the diffusion through the stratum corneum (SC). Therefore, it is expected that the development of a transdermally delivered formulation may enhance the permeability ratio (Qn) of ASI for its clinical application. In this study, we designed asiaticoside-loaded nanoemulsions (ASI-NEs) and nanoemulsions-based gels (ASI-NBGs) and studied their mechanism for transdermal delivery.Methods: The preparation of ASI-NEs was optimized by simplex lattice design (SLD). The ex vivo transdermal penetration and the in vivo pharmacokinetics studies were studied, respectively. The skin irritation of ASI-NEs and ASI-NBGs was measured on normal and damaged skin in rabbits, and the transcutaneous mechanisms of ASI-NEs and ASI-NBGs were determined by HE stained and confocal laser scanning microscopy (CLSM).Results: The mean particle size of ASI-NEs was 132± 5.84nm. The ex vivo skin permeation study verified that the Qn of the optimized ASI-NEs and ASI-NBGs was about 13.65 times and 5.05 times higher than that of the ordinary ASI-G group. In vivo, the pharmacokinetics studies showed that ASI-NEs and ASI-NBGs reached the peak value in the skin quickly and maintained stable release for a long time with high bioavailability. ASI-NEs and ASI-NBGs were proved to be safe when applied for topical skin usage, and they could play a therapeutic role through the skin mainly by acting on the microstructure of the SC and by means of the skin adnexal pathways.Conclusion: ASI-NEs and ASI-NBGs were effectively developed to overcome the barrier properties of the skin and show high drug penetration through the transdermal route. In addition, we found that ASI-NEs and ASI-NBGs are safe when applied through transdermal delivery system.Keywords: asiaticoside, transdermal delivery system, permeation enhancer, nanoemulsions

Qingquan Wei,1,* Zhongqi Wan,2,* Yongcheng Hu,3,* Qing Peng1 1Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University, Shanghai 200072, People’s Republic of China; 2School of Public Health, Nanjing Medical University, Nanjing 211166, Jiangsu Province, People’s Republic of China; 3Department of Ophthalmology, Bayannuer Paralympic Eye Hospital, Inner Mongolia 015000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Qing PengDepartment of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University, Shanghai 200072, People’s Republic of ChinaTel +86 13220811345Email pengqing@tongji.edu.cnPurpose: This study aimed to investigate the concentrations of cytokines and chemokines in diabetic macular edema (DME) eyes before and during therapy with the intravitreal injection of conbercept (IVC) and to identify associations with disease activity.Methods: The Bio-Plex® 200 System and the Bio-PlexTM Human Cytokine Standard 27-Plex, Group I (Bio-Rad, Hercules, California, USA) were used to detect cytokine levels in aqueous humour. Experimental aqueous humour samples were collected from 18 patients with DME at the same time that IVC was performed at baseline and at 1 month. Control aqueous humour samples were collected from 16 patients undergoing cataract surgery.Results: Significantly higher concentrations of vascular endothelial growth factor (VEGF), interleukin 6 (IL-6), IL-8, eotaxin, granulocyte colony stimulating factor (G-CSF), interferon gamma-induced protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) were found in the aqueous humour of DME patients than cataract patients. One month after IVC, the intraocular concentrations of VEGF were significantly lower in the eyes of DME patients than at baseline. No other cytokines were significantly altered by conbercept therapy. Best-corrected visual acuity (BCVA) slightly improved following IVC compared with that at baseline, although this difference was not significant, and central macular thickness (CMT) significantly decreased 1 month after IVC treatment.Conclusion: Angiogenic, inflammatory and growth factors are involved in the development of DME. With the exception of VEGF, IVC did not cause significant differences in any inflammatory cytokines or growth factors in DME patients. CMT is related to VEGF levels in aqueous humour.Keywords: diabetic macular oedema, conbercept, intravitreal implant, intraocular cytokines

Pengliang Xin,1 Wenqian Xu,1 Xiongpeng Zhu,1 Chuntuan Li,1 Yan Zheng,1 Tingjin Zheng,2 Wenzhao Cheng,3 Qunyi Peng11Department of Haematology, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, People’s Republic of China; 2Central Laboratory, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, People’s Republic of China; 3Stem Cell Translational Research Center, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, People’s Republic of ChinaCorrespondence: Xiongpeng ZhuQuanzhou First Hospital Affiliated to Fujian Medical University, No. 250 East Street, Licheng District, Quanzhou City, People’s Republic of ChinaTel +86 1 380 599 8581Email Xiongpengzhu@tom.comPurpose: To investigate the effects of BEZ235 on chronic myeloid leukemia (CML) cells.Methods: MTS assay was used to detect the proliferation of CML cells. The proteins expression were detected by Western blot assay. The effects of BEZ235 on autophagy in CML cells were verified through transmission electron microscopy and evaluated by laser confocal microscopy. Annexin V-FITC/PI double staining flow cytometry was used to detect apoptosis. A xenograft model was established to observe the therapeutic effect of BEZ235 in vivo.Results: BEZ235 could inhibit the proliferation of CML cells; CQ and 3-MA could increase the proliferation inhibition and Z-VAD-FMK can reduce the proliferation inhibition of BEZ235 on CML cells (P

Chunze Zhang,1,2,* Tingting Yin,3,* Ran Tao,4,* Bo Xiao,5,* Jing Chen,4 Zixuan Li,5 Xueyuan Miao,3 Qing Peng,3 Liu Sun,3 Weihua Zhang,1 Junxu Ren,4 Zhao Zhang,1 Ying Zhang,1 Xichuan Li,6 Wei Zhang51Department of Colorectal Surgery, Tianjin Union Medical Center, Tianjin, People’s Republic of China; 2State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, People’s Republic of China; 3Tianjin University of Traditional Chinese Medicine, Tianjin, People’s Republic of China; 4Department of Histology and Embryology, School of Basic Medical Sciences, Hebei North University, Zhangjiakou, Hebei, People’s Republic of China; 5School of Basic Medical Sciences, Tianjin Medical University, Tianjin, People’s Republic of China; 6Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, Tianjin, People’s Republic of China *These authors contributed equally to this work Purpose: Y-box binding protein 1 (YBX1) is a multifunctional protein linked to tumor progression and its elevated expression is an indicator of poor prognosis in various cancers. This meta-analysis aimed to investigate the prognostic value and clinical significance of YBX1 in malignant cancer.Methods: Relevant articles published through September 12, 2018 were identified from a comprehensive electronic and manual search in PubMed, Web of Science and Embase databases. The combined odds ratios (ORs) and hazard ratios (HRs) with 95% confidence intervals (95% CIs) were used to estimate the relationship among clinicopathological characteristics, overall survival and disease-free-survival of patients with solid tumor and YBX1 expression.Results: The study included 27 studies and 5,996 patients. Our analysis revealed significant association between increased YBX1 expression and tumor differentiation status, tumor size and lymph node metastasis; moreover, the pooled HR values demonstrated that high nuclear YBX1 expression was significantly associated with worse overall survival (HR=2.14; 95% CI: 1.72–2.67, P

Xueqing Zhang,1,* Jinluan Li,1,* Qingqin Peng,2 Yunxia Huang,1 Lirui Tang,1 Qingyang Zhuang,1 Feifei Lin,1 Xijin Lin,1 Kaixin Du,3 Junxin Wu1 1Department of Radiation Oncology, Fujian Medical University Cancer Hospital, Fujian Cancer Hospital, Fuzhou 350014, China; 2Department of Radiation Oncology, First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China; 3Department of Radiation Oncology, Xiamen Humanity Hospital, Xiamen 361000, China *These authors contributed equally to this work Purpose: The inflammatory status of patients with cancer appears to affect cancer progression and patient prognosis. We examined the characteristics of cancer-associated systemic and local inflammation and its impact on the overall survival (OS) of patients with locally advanced rectal cancer (LARC) who received neoadjuvant radiotherapy (nRT). Patients and methods: Seventy-six consecutive LARC patients who received nRT from February 2012 to September 2015 were retrospectively analyzed. The peripheral neutrophil-to-lymphocyte ratio (NLR) was determined at diagnosis, and the CD8+ T-cell count was determined from surgical specimens. Factors associated with OS were identified by univariate and multivariate Cox regression. Results: The median follow-up time was 23.0 months (range: 2–59), and the overall 5-year OS rate was 68.6% (95% CI =46.06–91.14). Patients with a high NLR (≥2.0) and a low CD8+ T-cell count (

Bozena Hosnedlova,1 Marta Kepinska,2 Sylvie Skalickova,3 Carlos Fernandez,4 Branislav Ruttkay-Nedecky,3 Qiuming Peng,5 Mojmir Baron,1 Magdalena Melcova,6 Radka Opatrilova,3 Jarmila Zidkova,6 Geir Bjørklund,7 Jiri Sochor,1 Rene Kizek2,3 1Department of Viticulture and Enology, Faculty of Horticulture, Mendel University in Brno, Lednice, Czech Republic; 2Department of Biomedical and Environmental Analyses, Faculty of Pharmacy, Wroclaw Medical University, Wroclaw, Poland; 3Central Laboratory, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic; 4School of Pharmacy and Life Sciences, Robert Gordon University, Aberdeen, UK; 5State Key Laboratory of Metastable Materials Science and Technology, Yanshan University, Qinhuangdao, People’s Republic of China; 6Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic; 7Council for Nutritional and Environmental Medicine, Rana, Norway Abstract: Traditional supplements of selenium generally have a low degree of absorption and increased toxicity. Therefore, it is imperative to develop innovative systems as transporters of selenium compounds, which would raise the bioavailability of this element and allow its controlled release in the organism. Nanoscale selenium has attracted a great interest as a food additive especially in individuals with selenium deficiency, but also as a therapeutic agent without significant side effects in medicine. This review is focused on the incorporation of nanotechnological applications, in particular exploring the possibilities of a more effective way of administration, especially in selenium-deficient organisms. In addition, this review summarizes the survey of knowledge on selenium nanoparticles, their biological effects in the organism, advantages, absorption mechanisms, and nanotechnological applications for peroral administration. Keywords: nanoparticles, biomedicine, drug delivery, oxidative stress, anticancer effect, antimicrobial activity, protective effect

Yu-Guo Yuan,1 Qiu-Ling Peng,2 Sangiliyandi Gurunathan3 1College of Veterinary Medicine/Animal Science and Technology/Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, People’s Republic of China; 2College of Chemistry and Bioengineering, Yichun University, Yichun, Jiangxi, People’s Republic of China; 3Department of Stem cell and Regenerative Biotechnology, Konkuk University, Seoul, Republic of Korea Background: Gemcitabine (GEM) is widely used as an anticancer agent in several types of solid tumors. Silver nanoparticles (AgNPs) possess unique cytotoxic features and can induce apoptosis in a variety of cancer cells. In this study, we investigated whether the combination of GEM and AgNPs can exert synergistic cytotoxic effects in the human ovarian cancer cell line A2780.Methods: We synthesized AgNPs using resveratrol as a reducing and stabilizing agent. The synthesized nanomaterials were characterized using various analytical techniques. The anticancer effects of a combined treatment with GEM and AgNPs were evaluated using a series of cellular assays. The expression of pro- and antiapoptotic genes was measured using real-time reverse transcription polymerase chain reaction. Apoptosis was confirmed by TUNEL assay.Results: In this study, combined treatment with GEM and AgNPs significantly inhibited viability and proliferation in A2780 cells. Moreover, the levels of apoptosis in cells treated with a combination of GEM and AgNPs were significantly higher compared with those in cells treated with GEM or AgNPs alone. Our data suggest that GEM and AgNPs exhibit potent apoptotic activity in human ovarian cancer cells. Combined treatment with GEM and AgNPs showed a significantly higher cytotoxic effect in ovarian cancer cells compared with that induced by either of these agents alone.Conclusion: Our study demonstrated that the interaction between GEM and AgNPs was cytotoxic in ovarian cancer cells. Combined treatment with GEM and AgNPs caused increased cytotoxicity and apoptosis in A2780 cells. This treatment may have therapeutic potential as targeted therapy for the treatment of ovarian cancer. To our knowledge, this study could provide evidence that AgNPs can enhance responsiveness to GEM in ovarian cancer cells and that AgNPs can potentially be used as chemosensitizing agents in ovarian cancer therapy. Keywords: cell viability, proliferation, cytotoxicity, apoptosis, gemcitabine, silver nanoparticles

Pengliang Xin, Chuntuan Li, Yan Zheng, Qunyi Peng, Huifang Xiao, Yuanling Huang, Xiongpeng Zhu Department of Haematology, First Hospital of Quanzhou Affiliated to Fujian Medical University, Licheng, Quanzhou, Fujian Province, China Background: Phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway is a therapy target of cancer. We aimed to confirm the effect of dual PI3K/mTOR inhibitor NVP-BEZ235 on proliferation, apoptosis, and autophagy of chronic myelogenous leukemia (CML) cells and sensitivity of tyrosine kinase inhibitor in vitro.Methods: Two human CML cell lines, K562 and KBM7R (T315I mutant strain), were used. The proliferation of CML cells was detected by MTS (Owen’s reagent) assay. Cell cycle and apoptosis assay were examined by flow cytometric analysis. The phosphorylation levels and the expression levels were both evaluated by Western blot analysis. NVP-BEZ235 in combination with imatinib was also used to reveal the effect on proliferation and apoptosis.Results: NVP-BEZ235 significantly inhibited the proliferation in a time- and dose-dependent manner, and the half-maximal inhibitory concentration values of NVP-BEZ235 inhibiting the proliferation of K562 and KBM7R were 0.37±0.21 and 0.43±0.27 µmol/L, respectively, after 48 h. Cell apoptosis assay showed that NVP-BEZ235 significantly increased the late apoptotic cells. Cell cycle analysis indicated that the cells were mostly arrested in G1/G0 phase after treatment by NVP-BEZ235. In addition, results also found that, after treatment by NVP-BEZ235, phosphorylation levels of Akt kinase and S6K kinase significantly reduced, and the expression levels of cleaved caspase-3 significantly increased; meanwhile, the expression levels of caspase-3, B-cell lymphoma-2, cyclin D1, and cyclin D2 significantly decreased, and the ratio of LC3II/LC3I was significantly increased with increased LC3II expression level. Moreover, imatinib in combination with NVP-BEZ235 induced a more pronounced colony growth inhibition than imatinib alone.Conclusion: NVP-BEZ235 effectively inhibited cell proliferation by G0/G1 cell cycle arrest and induced apoptosis through deregulating PI3K/Akt/mTOR pathway in CML cells; in addition, NVP-BEZ235 can enhance cell autophagy, and is conducive to raising CML cell sensitivity to imatinib to inhibit the growth of imatinib-resistant cells. Keywords: chronic myelogenous leukemia, NVP-BEZ235, phosphatidylinositol 3-kinase/Akt/mammalian pathway, imatinib, apoptosis, autophagy

Qing Peng,1 Albert K Dearden,2 Jared Crean,1 Liang Han,1 Sheng Liu,3 Xiaodong Wen,4,5 Suvranu De11Department of Mechanical, Aerospace and Nuclear Engineering, Rensselaer Polytechnic Institute, Troy, NY, USA; 2Department of Physics, Applied Physics, and Astronomy, Rensselaer Polytechnic Institute, Troy, NY, USA; 3Institute for Microsystems, School of Mechanical Engineering, Huazhong University of Science and Technology, Wuhan, People's Republic of China; 4State Key Laboratory of Coal Conversion, Institute of Coal Chemistry, Chinese Academy of Sciences, Taiyuan, People's Republic of China; 5Synfuels China Co, Ltd, Huairou, Beijing, People's Republic of ChinaAbstract: Plenty of new two-dimensional materials including graphyne, graphdiyne, graphone, and graphane have been proposed and unveiled after the discovery of the "wonder material" graphene. Graphyne and graphdiyne are two-dimensional carbon allotropes of graphene with honeycomb structures. Graphone and graphane are hydrogenated derivatives of graphene. The advanced and unique properties of these new materials make them highly promising for applications in next generation nanoelectronics. Here, we briefly review their properties, including structural, mechanical, physical, and chemical properties, as well as their synthesis and applications in nanotechnology. Graphyne is better than graphene in directional electronic properties and charge carriers. With a band gap and magnetism, graphone and graphane show important applications in nanoelectronics and spintronics. Because these materials are close to graphene and will play important roles in carbon-based electronic devices, they deserve further, careful, and thorough studies for nanotechnology applications.Keywords: two-dimensional materials, graphene-like structures, properties and synthesis, nanotechnology applications, graphyne, hydrogenation of grapheme

Ting Zhang,1 Yong Zheng,2 iang Peng,3 Xi Cao,1 Tao Gong,1 Zhirong Zhang11Key Laboratory of Drug Targeting and Drug Delivery Systems, Sichuan University, Chengdu, People’s Republic of China; 2Second Affiliated Hospital, Chongqing Medical University, Chongqing, People’s Republic of China; 3State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, People’s Republic of ChinaBackground: Vincristine (VCR), which is a widely used antineoplastic drug, was integrated with a submicron-emulsion drug-delivery system to enhance the anticancer effect.Methods: After the formation of a VCR–oleic acid ion-pair complex (VCR-OA), the VCR-OA-loaded submicron emulsion (VCR-OA-SME), prepared by classical high-pressure homogenization, was characterized and its in vitro anticancer effects were evaluated.Results: The submicron-emulsion formulation exhibited a homogeneous round shape. The mean particle size, zeta potential, and encapsulation efficiency were 157.6 ± 12.6 nm, −26.5 ± 5.0 mV and 78.64% ± 3.44%, respectively. An in vitro release study of the VCR-OA-SME revealed that 12.4% of the VCR was released within the first 2 hours (initial burst-release phase) and the rest of the drug was detected in the subsequent sustained-release phase. Compared with VCR solution, the pharmacokinetic study of VCR-OA-SME showed relatively longer mean residence time (mean residence time [0–∞] increased from 187.19 to 227.56 minutes), higher maximum concentration (from 252.13 ng/mL to 533.34 ng/mL), and greater area under the curve (area under the curve [0–∞] from 11,417.77 µg/L/minute to 17,164.34 µg/L/minute. Moreover, the VCR-OA-SME exhibited higher cytotoxicity (P < 0.05) on tumor cells by inducing cell arrest in the G2/M phase or even apoptosis (P < 0.05).Conclusion: The VCR-OA-SME formulation in our study displayed great potential for an anticancer effect for VCR.Keywords: ion-pair complex, submicron emulsion, cytotoxicity, apoptosis, cell uptake

Jinjie Zhang1, Qiang Peng1, Sanjun Shi1, Qiang Zhang2, Xun Sun1, Tao Gong1, Zhirong Zhang11Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu, People's Republic of China; 2State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Peking, People's Republic of ChinaBackground: As a poorly water-soluble drug, the oral application of morin is limited by its low oral bioavailability. In this study, a new self-nanoemulsifying drug delivery system (SNEDDS), based on the phospholipid complex technique, was developed to improve the oral bioavailability of morin.Methods: Morin-phospholipid complex (MPC) was prepared by a solvent evaporation method and characterized by infrared spectroscopy and X-ray diffraction. After formation of MPC, it was found that the liposolubility of morin was significantly increased, as verified through solubility studies. An orthogonal design was employed to screen the blank SNEDDS, using emulsifying rate and particle size as evaluation indices. Ternary phase diagrams were then constructed to investigate the effects of drug loading on the self-emulsifying performance of the optimized blank SNEDDS. Subsequently, in vivo pharmacokinetic parameters of the morin-phospholipid complex self-nanoemulsifying drug delivery system (MPC-SNEDDS) were investigated in Wistar rats (200 mg/kg of morin by oral administration).Results: The optimum formulation was composed of Labrafil® M 1944 CS, Cremophor® RH 40, and Transcutol® P (3:5:3, w/w), which gave a mean particle size of approximately 140 nm. Oral delivery of the MPC-SNEDDS exhibited a significantly greater Cmax (28.60 µg/mL) than the morin suspension (5.53 µg/mL) or MPC suspension (23.74 µg/mL) (all P < 0.05). Tmax was prolonged from 0.48 to 0.77 hours and to 1 hour for MPC and MPC-SNEDDS, respectively. In addition, the relative oral bioavailability of morin formulated in the MPC-SNEDDS was 6.23-fold higher than that of the morin suspension, and was significantly higher than that of the MPC suspension (P < 0.05).Conclusion: The study demonstrated that a SNEDDS combined with the phospholipid complex technique was a promising strategy to enhance the oral bioavailability of morin.Keywords: morin, phospholipid complex, self-nanoemulsifying drug delivery system, oral bioavailability

The present study aimed at detecting the association of ovine major histocompatibility complex class II (Ovar II) DRB1 gene second exon and susceptibility or resistance to hydatidosis in three sheep breeds of Sinkiang. The MHC-DRB1 second exon was amplified by polymerase chain reaction (PCR) from DNA samples of healthy sheep and sheep with hydatidosis. PCR products were characterized by the restriction fragment length polymorphism (RFLP) technique. Five restriction enzymes, MvaI, HaeIII, SacI, SacII, Hin1I, were used, yielding 14 alleles and 31 restriction patterns. Frequencies of patterns MvaIbc, Hin1Iab, SacIIab, HaeIIIde, HaeIIIdf, HaeIIIdd (P < 0.01) in Kazakh sheep, SacIab (P < 0.05) in Duolang sheep, and HaeIIIab, HaeIIIce, HaeIIIde, HaeIIIee (P < 0.01) in Chinese Merino (Sinkiang Junken type) sheep, were significantly higher in healthy sheep compared with infected sheep. These results indicated a strong association between these patterns and hydatidosis resistance. In contrast, the frequencies of MvaIbb, SacIIaa, Hin1Ibb, HaeIIIef (P < 0.01) and HaeIIIab (P < 0.05) in Kazakh sheep, SacIbb, HaeIIIae, Hin1Iab (P < 0.05), HaeIIIaa, HaeIIIbe, HaeIIIef (P < 0.01) in Duolang sheep, SacIIaa (P < 0.05) and HaeIIIbd, Hin1Ibb, HaeIIIcf, HaeIIIef (P < 0.01) in Chinese Merino sheep (Sinkiang Junken type) were significantly lower in healthy sheep compared with infected sheep. This indicated a strong association between these patterns and hydatidosis susceptibility. In addition, sheep with the pattern of HaeIIIef demonstrated a high hydatidosis susceptibility (P < 0.01) in all three breeds, while sheep with the pattern HaeIIIde demonstrated significant hydatidosis resistance (P < 0.01) in Kazakh and Chinese Merino sheep (Sinkiang Junken type). These results suggest that the Ovar-DRB1 gene plays a role in resistance to hydatidosis infection in the three sheep breeds.