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Ming-Ming Li,* Yi Zhang,* Fang Sun, Man-Xiu Huai, Fei-Yu Zhang, Jia-Xing Pan, Chun-Ying Qu, Feng Shen, Zheng-Hong Li, Lei-Ming Xu Department of Gastroenterology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Lei-Ming Xu; Yi Zhang, Department of Gastroenterology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, 1665 Kongjiang Road, Shanghai, People’s Republic of China, 200092, Tel/Fax +86 21 2507 8999, Email xuleiming@xinhuamed.com.cn; zhangyi02@xinhuamed.com.cnPurpose: Photodynamic therapy (PDT) induces anti-tumor immune responses by triggering immunogenic cell death in tumor cells. Previously, we demonstrated that novel QDs-RGD nanoparticles exhibited high efficiency as photosensitizers in the treatment of pancreatic cancer. However, the underlying mechanism of the anti-tumor immune effects induced by the photosensitizer remains unknown. This study assessed the anticancer immune effect of QDs-RGD, as well as the conventional photosensitizer chlorine derivative, YLG-1, for comparison, against pancreatic cancer in support of superior therapeutic efficacy.Methods: The pancreatic cancer cell line, Panc02, was used for in vitro studies. C57BL/6 mice bearing pancreatic cancer cell-derived xenografts were generated for in vivo studies to assess the anti-tumor effects of QDs-RGD-PDT and YLG-1-PDT. The immunostimulatory ability of both photosensitizers was examined by measuring the expression of damage-associated molecular patterns (DAMP), such as calreticulin (CRT), assessing dendritic cell (DC) maturation, and analyzing cytokine expression. The specific immunity of QDs-RGD and YLG-1-PDT on distant tumor were determined by combining PDT with anti-CTLA-4 antibody.Results: QDs-RGD-PDT and YLG-1-PDT significantly inhibited pancreatic cancer cell growth in a dose- and time-dependent manner. While both photosensitizers significantly promoted CRT release, DC maturation, and interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α) expression, QDs-RGD exerted a stronger immunostimulatory effect than YLG-1. Combination treatment with QDs-RGD and CTLA-4 blockade was able to significantly inhibit the growth of distant tumors.Conclusion: QDs-RGD is a novel and effective PDT strategy for treating pancreatic tumors by inducing anti-tumor immune responses.Keywords: quantum dots, RGD peptides, cancer immunotherapy, pancreatic neoplasm

Qingqing Luo,1,* Yibo Quan,1,* Wei Liu,1 Zixin Wu,1 Wenjing Qiu,1 Wenlong Liang,1 Ping Yang,1 Qing Huang,1 Guanwei Li,1 Jianchang Wei,1 Qiang Wang,1 Fei Shen,2 Wanglin Li,1 Feng He,3 Jie Cao1 1Department of Gastrointestinal Surgery, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, 510180, People’s Republic of China; 2Department of Thyroid Surgery, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, 510180, People’s Republic of China; 3Department of Nephrology, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, 510180, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jie Cao, Department of Gastrointestinal Surgery, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, 510180, People’s Republic of China, Email eycaojie@scut.edu.cn Feng He, Department of Nephrology, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, 510180, People’s Republic of China, Email eyhefeng@scut.edu.cnPurpose: Consensus molecular subtypes (CMS) are mainly used for biological interpretability and clinical stratification of colorectal cancer (CRC) in primary tumors (PT) but few in metastases. The heterogeneity of CMS distribution in metastases and the concordance of CMS between PT and metastases still lack sufficient study. We used CMS to classify CRC metastases and combine it with histopathological analysis to explore differences between PT and distant metastases.Patients and Methods: We obtained gene expression profiles for 942 PT samples from TCGA database (n=376) and GEO database (n=566), as well as 442 metastasis samples from GEO database. Among these, 765 PT samples and 442 metastasis samples were confidently identified with CMS using the “CMS classifier” and enrolled for analysis. Clinicopathological manifestation and CMS classification of CRC metastases were assessed with data from GEO, TCGA, and cBioPortal. Overall, 105 PT-metastasis pairs were extracted from 10 GEO datasets to assess CMS concordance. Tumor microenvironment (TME) features between PT and metastases were analyzed by immune-stromal infiltration with ESTIMATE and xCell algorithms. Finally, TME features were validated with multiplex immunohistochemistry in 27 PT-metastasis pairs we retrospectively collected.Results: Up to 64% of CRC metastases exhibited concordant CMS groups with matched PT, and the TME of metastases was similar to that of PT. For most common distant metastases, liver metastases were predominantly CMS2 and lung and peritoneal metastases were mainly CMS4, highlighting “seed” of tumor cells of different CMS groups had a preference for metastasis to “soil” of specific organs. Compared with PT, cancer-associated fibroblasts (CAF) reduced in liver metastases, CD4+T cells and M2-like macrophages increased in lung metastases, and M2-like macrophages and CAF increased in peritoneal metastases.Conclusion: Our findings underscore the importance of CMS-guided specific organ monitoring and treatment post-primary tumor surgery for patients. Differences in immune-stromal infiltration among different metastases provide targeted therapeutic opportunities for metastatic CRC.Keywords: colorectal cancer, primary tumors, metastases, consensus molecular subtypes, tumor microenvironment

Shen F, Pan X, Li M, Chen Y, Jiang Y, He J. Onco Targets Ther. 2020;13:3165–3176. We, the Editor and Publisher of the journal OncoTargets and Therapy have retracted the published article. Following publication of the article, the authors raised concerns about the duplication of images from Figures 5 and 6. Specifically, The images for Figure 5B, Veh and Nec-1, 0h, have been duplicated. The images for Figure 6A and 6B, MCF-7, Nec-1, have been duplicated. The authors were cooperative and were able to provide some original data for the study but were unable to provide a satisfactory explanation for how the images came to be duplicated. As verifying the validity of published work is core to the integrity of the scholarly record, the Publisher and Editor requested to retract the article and the authors do not agree with this decision. We have been informed in our decision-making by our editorial policies and COPE guidelines. The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as “Retracted”.

Cong Zhou,1 Hui Zhang,1 Maosuo Xu,1 Yajuan Liu,1 Baoyu Yuan,2 Yong Lin,1 Fang Shen1 1Department of Clinical Laboratory, Shanghai Fifth People’s Hospital, Fudan University, Shanghai, People’s Republic of China; 2Department of Clinical Laboratory, Shanghai Children’s Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of ChinaCorrespondence: Fang Shen; Yong Lin, Department of Clinical Laboratory, Shanghai Fifth People’s Hospital, Fudan University, No. 128, Ruili Road, Minhang District, Shanghai, 200240, People’s Republic of China, Tel +86 18021073261 ; +86 18718168695, Email shenfang5th@aliyun.com; linyong7007@163.comBackground: Hypervirulent carbapenem-resistant Klebsiella pneumoniae (hv-CRKP) has recently aroused an extremely severe health challenge and public concern. However, the underlying mechanisms of fitness costs that accompany antibiotic resistance acquisition remain largely unexplored. Here, we report a hv-CRKP-associated fatal infection and reveal a reduction in virulence due to the acquisition of aminoglycoside resistance.Methods: The bacterial identification, antimicrobial susceptibility, hypermucoviscosity, virulence factors, MLST and serotypes were profiled.The clonal homology and plasmid acquisition among hv-CRKP strains were detected by XbaI and S1-PFGE. The virulence potential of the strains was evaluated using Galleria mellonella larvae infection model, serum resistance assay, capsular polysaccharide quantification, and biofilm formation assay. Genomic variations were identified using whole-genome sequencing (WGS).Results: Four K. pneumoniae carbapenemase (KPC)-producing CRKP strains were consecutively isolated from an 86-year-old patient with severe pneumonia. Whole-genome sequencing (WGS) showed that all four hv-CRKP strains belonged to the ST11-KL64 clone. PFGE analysis revealed that the four ST11-KL64 hv-CRKP strains could be grouped into the same PFGE type. Under the pressure of antibiotics, the antimicrobial resistance of the strains increased and the virulence potential decreased. Further sequencing, using the Nanopore platform, was performed on three representative isolates (WYKP586, WYKP589, and WYKP594). Genomic analysis showed that the plasmids of these three strains underwent a large number of breaks and recombination events under antibiotic pressure. We found that as aminoglycoside resistance emerged via acquisition of the rmtB gene, the hypermucoviscosity and virulence of the strains decreased because of internal mutations in the rmpA and rmpA2 genes.Conclusion: This study shows that ST11-KL64 hv-CRKP can further evolve to acquire aminoglycoside resistance accompanied by decreased virulence to adapt to antibiotic pressure in the host.Keywords: hypervirulent carbapenem-resistant Klebsiella pneumoniae, within-host evolution, plasmids, whole-genome sequencing

Fei Shen, Jihe Huang, Kejia Yang, Chunhua Sun Department of Orthopedics, Suzhou Wuzhong People’s Hospital, Suzhou, People’s Republic of ChinaCorrespondence: Fei Shen; Chunhua Sun, Department of Orthopedics, Suzhou Wuzhong People’s Hospital, No. 61 Dongwu North Road, Wuzhong District, Suzhou, Jiangsu Province, 215128, People’s Republic of China, Email sanfi2000@sina.com; schaofie2023@163.comAbstract: Bone metastasis is one of the most important factors associated with poor prognosis for patients with prostate, breast, thyroid, and lung cancer. In the past two decades, 651 clinical trials, including 554 interventional trials, were being registered in ClinicalTrials.gov and pharma.id.informa.com to combat bone metastases from different perspectives. In this review, we comprehensively analyzed, regrouped, and discussed all the interventional trials on bone metastases. Clinical trials were re-grouped into bone-targeting agents, radiotherapy, small molecule targeted therapy, combination therapy, and others, based on the different mechanisms of action including modifying the bone microenvironment and preventing the growth of cancer cells. We also discussed the potential strategies that might improve overall survival and progression-free survival of patients with bone metastases in the future.Keywords: bone metastases, clinical trials, interventional, ClinicalTrials.gov, pharma.id.informa.com

Yeping Song,1 Fei Shen,1 Qing Dong,1 Liling Wang,1,* Jianhua Mi2,* 1Cerebrovascular Disease Center, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, 200127, People’s Republic of China; 2Health Management Center, Renji Hospital, School of Medical School, Shanghai Jiaotong University, Shanghai, 200127, People’s Republic of China*These authors contributed equally to this workCorrespondence: Liling Wang; Jianhua Mi, Email drliling0911@163.com; mjh851123@sina.comPurpose: Among all ischemic stroke patients, more than half are mild and rapidly improving acute ischemic stroke (MaRAIS) patients. However, many MaRAIS patients do not recognize the disease early on, and thus they delay access to the treatment that would be most effective if provided earlier. This is especially true in rural areas. The aim of this study was to develop and validate a late hospital arrival risk nomogram in a rural Chinese population of patients with MaRAIS.Methods: We developed a prediction model based on a training dataset of 173 MaRAIS patients collected from September 9, 2019 to May 13, 2020. Data analyzed included demographics and disease characteristics. A least absolute shrinkage and selection operator (LASSO) regression model was used to optimize feature selection for the late hospital arrival risk model. Multivariable logistic regression analysis was applied to build a prediction model incorporating the features selected in the LASSO regression models. The discrimination, calibration, and clinical usefulness of the prediction model were assessed using the C-index, calibration plot, and decision curve analysis, respectively. Internal validation was then assessed using bootstrapping validation.Results: Variables contained in the prediction nomogram included transportation mode, history of diabetes, knowledge of stroke symptoms, and thrombolytic therapy. The model had moderate predictive power with a C-index of 0.709 (95% confidence interval: 0.636– 0.783) and good calibration. In the internal validation, the C-index reached 0.692. The risk threshold was 30– 97% according to the analysis of the decision curve, and the nomogram could be applied in clinical practice.Conclusion: This novel nomogram, which incorporates transportation mode, history of diabetes, knowledge of stroke symptoms, and thrombolytic therapy, was conveniently applied to facilitate individual late hospital arrival risk prediction among MaRAIS patients in a rural area of Shanghai, China.Keywords: mild and rapidly improving acute ischemic stroke, mild stroke, prehospital delay, late hospital arrival

The regions of interaction between solar wind streams of different speed, known as corotating interaction regions, form an almost constantly existing structure of the inner heliosphere. Using observational data on the main characteristics of the heliosphere, important for GCR modulation, and the results of 3D MHD modeling of corotating interaction regions, and Monte Carlo simulation of recurrent GCR variations, we analyze the importance of the corotating interaction regions for longitude-averaged characteristics of the heliosphere and GCR propagation, and possible ways for simulating long-term GCR intensity variations with respect to the corotating interaction regions.

Jian Peng,1– 3,* Yue Wang,3,* Zhaoyin Wu,3 Chengju Mao,3 Lu Li,1,2 Huijun Cao,4 Zhilang Qiu,3 Guo Guo,3,5 Guiyou Liang,4,5 Feng Shen1,2 1Department of Intensive Care Unit, Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, 550009, People’s Republic of China; 2Guizhou Institute of Precision Medicine, Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, 550009, People’s Republic of China; 3Key Laboratory of Infectious Immune and Antibody Engineering of Guizhou Province, Cellular Immunotherapy Engineering Research Center of Guizhou Province, School of Biology and Engineering/School of Basic Medical Sciences, Guizhou Medical University, Guiyang, Guizhou, 550025, People’s Republic of China; 4Department of Cardiac Surgery, the affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, 550009, People’s Republic of China; 5Translational Medicine Research Center, Guizhou Medical University, Guiyang, Guizhou, 550025, People’s Republic of China*These authors contributed equally to this workCorrespondence: Feng Shen; Guiyou Liang, Email doctorshenfeng@163.com; guiyou515@163.comIntroduction: Acinetobacter baumannii has become a major difficulty in the treatment of bacteria-associated infection. The previously reported antimicrobial peptide Cec4 exhibited good and stable activity against A. baumannii in vitro, but the mechanisms and effects in vivo are elusive.Methods: The effects of Cec4 on bacterial membrane permeability, membrane potential and bacterial reactive oxygen species were measured. The cell membrane localization of antimicrobial peptides was studied by fluorescence labelling. The ability of bacteria to develop resistance to antimicrobial peptides was studied by continuous induction, and transcriptome difference was analysed. The in vivo toxicity of Cec4 against nematodes and mice was studied, and the in vivo therapeutic potential of Cec4 against A. baumannii was assessed.Results: Cec4 effectively cleared multidrug-resistant A. baumannii by altering bacterial cell membrane permeability, changing bacterial cell membrane polarity, and increasing bacterial intracellular reactive oxygen species. Cec4 affected the expression of the secretion system, outer membrane, and efflux pump genes of A. baumannii. In addition, the bacteria did not acquire stable drug-resistant ability. Cec4 at 1.024 mg/mL did not affect the proliferation of HeLa and HepG2 cells, and Cec4 at 45 mg/kg had little effect on the mortality of Caenorhabditis elegans, even the liver and kidney tissues of mouse. Most importantly, Cec4 could effectively improve the survival rates and reduce the bacterial load of various tissues in the mouse model of infection.Conclusion: In conclusion, Cec4 can damage the cell membrane of bacteria, and the bacteria is not easy to produce resistance to Cec4. Besides, Cec4 has good potential for the treatment of multidrug-resistant A. baumannii infections.Keywords: antimicrobial peptide, A. baumannii, sepsis, skin infection, drug safety

Mudan He,1,* Guoqing Zhang,2,* Fang Shen,1 Xingjing Li1 1Department of Respiratory, Zhongshan Hospital Wusong Branch, Fudan University, Shanghai, 201900, People’s Republic of China; 2Department of Respiratory, Jiading Branch of Shanghai General Hospital, Shanghai Jiao tong University School of Medicine, Shanghai, 200803, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xingjing Li, Department of Respiratory, Zhongshan Hospital Wusong Branch, Fudan University, No. 101 of North Tongtai Road, Baoshan District, Shanghai, 201900, People’s Republic of China, Tel +86 13816446543, Email lixingjinglxj9a@126.comBackground and Objectives: Smoking can lead to airway inflammation and mucus secretion through the nucleotide-binding domain-like receptor protein 3/caspase-1 pathway. In this study, z-VaD-Ala-Asp-fluoromethyl ketone(Z-VAD), a pan-caspase inhibitor, and acetyl-Asp-Glu-Val-Asp-aldehyde(Ac-DEVD), a caspase-3 inhibitor, were used to investigate the effect of caspase inhibitors on the expression of interleukin(IL)-1β and IL-8, airway inflammation, and mucus secretion in mice exposed to cigarette smoke(CS).Methods: Thirty-two C57BL/6J male mice were divided into a control group, Smoke group, Z-VAD group, and Ac-DEVD group. Except for the control group, the animals were all exposed to CS for three months. After the experiment, lung function was measured and hematoxylin and eosin staining and periodic acid–Schiff staining were performed. The levels of IL-1β, IL-8, and mucin 5ac(Muc5ac) in serum and bronchoalveolar lavage fluid(BALF) were determined by enzyme-linked immunosorbent assay.Results: Compared with the control group, the lung function of mice exposed to smoke was poorer, with a large number of inflammatory cells infiltrating around the airway, collapse of alveoli, expansion and fusion of distal alveoli, and formation of emphysema. The Z-VAD group was relieved compared with the smoke group. Airway inflammation was also reduced in the Ac-DEVD group compared with the Smoke group, but the degree of emphysema was not significantly improved. Although Z-VAD relieved airway inflammation and emphysema, Ac-DEVD only relieved inflammation. Z-VAD and Ac-DEVD decreased serum IL-1β and IL-8 levels. In BALF, IL-1β was decreased in Z-VAD group and IL-8 was highest in Smoke +Ac-DEVD group compared with control group and Ac-DEVD group. There was no significant difference in the expression of Muc5ac in serum. However, in BALF, levels of Muc5ac were higher in the smoking group and the lowest in the Ac-DEVD group.Conclusion: Mice exposed to smoke had decreased lung function and significant cilia lodging, epithelial cell shedding, and inflammatory cell infiltration, with significant emphysema formation. The pan-caspase inhibitor, Z-VAD, improved airway inflammation and emphysema lesions in the mice exposed to smoke and reduced IL-1β and IL-8 levels in serum. The caspase-3 inhibitor, Ac-DEVD, reduced airway inflammation, serum IL-1β and IL-8 levels, and Muc5ac levels in BALF, but it did not improve emphysema.Keywords: cigarette smoke, Z-VAD-FMK, Ac-DEVD-CHO, inflammation, mucin 5ac

With a view to understanding the performance of the blast furnace slag with high Al2O3 content, the effects of w(MgO)/w(Al2O3), w(CaO)/w(SiO2), and w(Al2O3) on the melting performance (melting characteristic temperature and melting heat) of the blast furnace slag with high Al2O3 content were investigated by the differential scanning calorimeter (DSC) method. Experimental results indicate that melting end temperature (Tend) for almost all the slags has no obvious change with the increase of w(MgO)/w(Al2O3), w(CaO)/w(SiO2) and w(Al2O3) of the slag will raise Tend of the slag. When (w(MgO)/w(Al2O3), R, and w(Al2O3) are high, the melting onset temperature (Tonset) of the slag increases with the increase of any variables. When w(CaO)/w(SiO2) is low, Tonset of the slag decreases with the increase of w(Al2O3). w(MgO)/w(Al2O3), w(CaO)/w(SiO2), and w(Al2O3) within the scope of this study and all these factors lead to the increase of the slag melting heat.

Kejun Peng,1,* Nianhua Deng,1,* Yao Meng,1,* Qianchuan He,2 Hao Meng,3 Ting Luo,4 Yanru Wei,1 Yue Kang,1 Xiaodong Zhou,1 Fubing Shen1 1School of Laboratory Medicine, Chengdu Medical College, Chengdu, People’s Republic of China; 2Public Health Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, 98109, USA; 3PKU-IDG/McGovern Institute for Brain Research, Peking-Tsinghua Center for Life Sciences, Peking University School of Life Sciences, Beijing, People’s Republic of China; 4Wuhan Corebiolab Co., Ltd, Wuhan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Fubing Shen, School of Laboratory Medicine, Chengdu Medical College, No. 783 Xindu Avenue, Xindu District, Chengdu, 610500, People’s Republic of China, Tel +86 28 62739522, Email shenfubing@163.comBackground: Alpha-momorcharin (α-MMC) is a natural medicine derived from bitter melon and has been found to exert immunomodulatory effects. Our previous study indicated that α-MMC can regulate cytokine release from monocytes, but it remains unknown about its regulatory effect on different types of cytokines, such as inflammatory cytokines or anti-inflammatory cytokines.Methods: LPS-induced M1-type macrophages model and IL-4-induced M2-type macrophages model were established, and the expression of proinflammatory cytokines and anti-inflammatory cytokines were assessed by ELISA after α-MMC was administered. Then, a LPS-induced acute pneumonia mouse model was established, the proinflammatory cytokines levels and inflammatory lesions in lung tissues were examined by ELISA or H&E staining. Furthermore, omics screening analysis and Western blotting verification were performed on TLR4 and JAK1-STAT6 signalling pathway-related proteins to elucidate the regulatory mechanism of α-MMC in those M1 macrophages and M2 macrophages.Results: At a noncytotoxic dose of 0.3 μg/mL, α-MMC significantly inhibited the LPS-induced expression of inflammatory cytokines, such as TNF-α, IL-1β, IL-6, IL-8, MIP-1α and MCP-1, by M1 macrophages in a time-dependent manner, but α-MMC did not inhibit the IL-4-induced synthesis of anti-inflammatory cytokines, such as IL-10, IL-1RA, EGF, VEGF, TGF-β and CCL22, by M2 macrophages. Moreover, α-MMC also inhibited inflammatory cytokine expression in an LPS-induced acute pneumonia mouse model and alleviated inflammation in lung tissues. Furthermore, omics screening and Western blotting analysis confirmed that α-MMC inhibited TAK1/p-TAK1 and subsequently blocked the downstream MAPK and NF-κB pathways, thus inhibiting the LPS-induced inflammatory cytokine expression.Conclusion: Our results reveal that α-MMC inhibits proinflammatory cytokine expression by M1 macrophages but not anti-inflammatory cytokine expression by M2 macrophages. The efficacy of α-MMC in selectively inhibiting proinflammatory cytokine expression renders it particularly suitable for the treatment of severe inflammation and autoimmune diseases characterized by cytokine storms.Keywords: α-MMC, M1 macrophage, proinflammatory cytokine, TLR4-NF-κB/MAPK

Liming Fan,1,&ast; Deyang Sun,1,&ast; Jia Yang,1 Xiawei Shi,1 Fenglin Shen,1 Ke Chen,1 Junchao Yang1,2 1The First Clinical College, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 2Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Junchao Yang, Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University, 54 Youdian Road, Shangcheng District, Hangzhou City, Zhejiang Province, People’s Republic of China, Tel +86-13858036093, Email yangjunchaozj@zcmu.edu.cnPurpose: The purpose of our study was to investigate the relationship between serum sodium levels and 1-year and 3-year mortality in critically ill patients with comorbid chronic obstructive pulmonary disease using real-world data.Methods: The data of this study were collected from the Medical Information Mart for Intensive Care-IV (MIMIC-IV) database. First of all, we used the Kaplan–Meier curves and multivariable Cox regression analyses to measure the relationship between serum sodium levels and 1-year and 3-year mortality for critically ill patients with comorbid COPD. Next, a restricted cubic spline was used to analyze non-parametrically the relationship between mortality and serum sodium as a continuous variable. In addition, we also analyzed the mortality of different subgroups.Results: A total of 5540 eligible subjects were extracted. Compared to normal serum sodium levels, adjusted multivariable Cox regression analysis confirmed that hyponatremia and hypernatremia were still significantly associated with 1-year mortality (HR = 1.551, 95% CI = 1.333∼ 1.805, P< 0.001; HR = 1.683, 95% CI = 1.317∼ 2.151, P< 0.001, respectively) and 3-year mortality (HR = 1.507, 95% CI = 1.302∼ 1.744, P< 0.001; HR = 1.612, 95% CI = 1.269∼ 2.048, P< 0.001, respectively). In patients with or without adjustment variables, there was an obvious U-shaped non-linear relationship between serum sodium levels and 1-year and 3-year mortality with a reference level of 139 mmol/L, which indicated that patients in both hyponatremia and hypernatremia had higher mortality than normal serum sodium levels.Conclusion: This study showed that both hyponatremia and hypernatremia were related to increased 1-year and 3-year mortality in critically ill patients with comorbid COPD, which provides a new reference for the control strategy of correcting serum sodium levels.Keywords: serum sodium, chronic obstructive pulmonary disease, intensive care unit, MIMIC-IV

Jian Fan,1,&ast; Yan Shang,1,&ast; Fei Shen,2,&ast; Qiaowen Huang,3 Ziyu Wang,1 Yiping Han,4 Jiayi Zhao1 1Department of General Practice, Changhai Hospital, Naval Military Medical University, Shanghai, 200433, People’s Republic of China; 2Yinhang Community Healthcare Center, Shanghai, 200433, People’s Republic of China; 3Zhaoxiang Community Healthcare Center, Shanghai, 200433, People’s Republic of China; 4Department of Respiratory and Critical Care Medicine, Changhai Hospital, Naval Military Medical University, Shanghai, 200433, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jiayi Zhao, Department of General Practice, Changhai Hongkou Hospital, Shanghai, 200081, People’s Republic of China, Email rayzhaojiayi@163.com Yiping Han, Department of Respiratory and Critical Care Medicine, Changhai Hospital, Naval Military Medical University, Shanghai, 200433, People’s Republic of China, Email yphan2006@163.comBackground: Venous thrombosis (VTE) is a high-incidence disease that affects different systems of the body, but no comparative studies have been conducted to explore its incidence in different systems in patients with the coexistence of multiple diseases. Primary care is a medical institution that has the first contact with patients’ conditions. The majority of outpatients suffer from multiple diseases, and they are more prone to VTE, which is of significance and requires investigation. At the same time, we observed that the primary care patients’ cognition level of VTE is insufficiency at present, and the prevention and management of VTE in China is still in its infancy. Most studies focus on whether doctors and nurses are clear about the diagnosis and manifestations of VTE, however, few studies on patients’ cognitive level and influencing factors of symptoms and prevention of VTE. Therefore, it is necessary to study the causes of cognitive deficiency of VTE from the perspective of patients, conduct targeted cognitive and behavioral interventions, and finally achieve the purpose of improving the effect of VTE prevention and management.Aim: This study aimed to explore the risk factors, cognitive level and cognitive potential influence factors of VTE in outpatients of primary care in Shanghai, as well as which diseases, are more likely to promote occurrence of VTE among patients with multiple diseases.Methods: A total of 710 patients were included in the analysis. Shanghai in China is divided into urban and suburban areas by their geographical location, and a random sample of 10 primary care areas was investigated, including 5 urban areas and 5 suburban areas. The Padua scale and the VTE cognitive level-related knowledge test scale were used for the investigation. Statistical software was used to conduct univariate Pearson correlation analysis and multivariate logistic regression analysis for VTE high-risk factors among outpatients of primary care. Furthermore, the high-risk factors for VTE and the types of chronic disease in patients with multiple diseases were subjected to subgroup analyses. t-test was used to statistically analyze cognitive level of VTE and the potential influence factors related to the patients’ VTE cognitive level.Results: Univariate analysis of risk factors for VTE in 710 outpatients showed that age (χ2=83.895, P< 0.001), education (χ2=8.324, P=0.004), region (χ2=63.936, P< 0.001), chronic disease (χ2=18.198, P< 0.001) and the coexistence of multiple diseases (χ2=61.347, P< 0.001) were the main risk factors for VTE risk in patients. Logistic multivariate regression analysis showed that age, region, chronic diseases and the coexistence of multiple diseases were independent risk factors for VTE risk in patients. The correlation between high risk of VTE in patients with multiple diseases and various types of chronic diseases was further analyzed by logistic multivariate regression. The results showed that diabetes mellitus (OR=1.636 [95% CI, 1.030– 2.599], P=0.037), cerebral infarction (OR=8.484 [95% CI, 4.615– 15.597], P< 0.001), coronary heart disease (OR=2.987 [95% CI, 1.699– 5.252], P< 0.001), intracerebral hemorrhage (OR=15.130 [95% CI, 4.707– 48.630], P< 0.001), respiratory diseases (OR=3.470, [95% CI, 1.428– 8.43], P=0.006) and malignant tumors (OR=4.576, [95% CI, 1.85– 11.315], P=0.001) were independent risk factors for VTE among patients with co-existing diseases, and the risks from high to low were cerebral hemorrhage>cerebral infarction>malignant tumor>respiratory system disease>coronary heart disease> diabetes. In addition, patients’ age (t=6.869, P< 0.001), region (t=3.420, P< 0.001), education level (t=− 7.341, P< 0.001), chronic disease (t=5.875, P< 0.001), BMI (t=6.404, P=0.012), smoking (t=4.152, P=0.042) were related to the VTE cognitive level.Conclusion: General practitioners should pay close attention to independent risk factors for VTE among patients in routine outpatient care, especially patients with co-existing diseases with independent risk for VTE, placing them high risk of VTE. At the same time, health education and other measures should be strengthened to improve the patients’ awareness and cognitive level of VTE to prevent the occurrence of VTE and its related complications.Keywords: venous thrombosis, risk assessment, cognitive level

Yiying Liu,1,&ast; Xin Wang,2,&ast; Linying Wang,1 Wenwen Chen,3 Wenyue Liu,1 Tingting Ye,1 Jing Hong,1 Hong Zhu,1 Feixia Shen1 1Department of Endocrinology and Metabolism, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 2Department of Urology, Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University, Taizhou, People’s Republic of China; 3Department of Senile Diseases, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Feixia ShenDepartment of Endocrinology and Metabolism, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of ChinaEmail shenfeixia@wmu.edu.cnPurpose: Diabetic neurogenic bladder (DNB) has been widely recognized in recent years. It is common in patients with long-term diabetes and may also lead to many severe complications. Although there has been widespread evidence that inflammation is involved in the development of some diabetic complications, there is little evidence that this can also occur in the bladder. In recent years, platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) have been viewed as potential novel markers of inflammatory responses. This study was designed to evaluate the relationship between the presence of DNB and the PLR and NLR.Patients and Methods: A total of 371 cases of T2DM patients were included in this retrospective study. Patients were divided into two groups, with 115 diabetic subjects diagnosed with diabetic neurogenic bladder and 256 control subjects without DNB. The independent predictors of DNB were analyzed using logistic regression.Results: Compared with patients without DNB, the mean PLR and NLR were significantly higher in those with DNB (p < 0.001). Based on the logistic regression, PLR was found to be an independent risk factor for DNB (odds ratio [OR]: 1.408, 95% confidence interval [CI]: 1.248– 1.617). From the receiver operating characteristic (ROC) curve, using PLR as indicative of DNB was expected to be 101.1949, and it generated a sensitivity and specificity value of 89.6% and 23.4%, respectively. The area under the curve (AUC) was also found to be 0.899 (95% CI: 0.865– 0.932).Conclusion: In our study, PLR and NLR were significantly higher for patients with DNB. The PLR was found to be a risk factor in the presence of DNB after correcting for possible confounding factors. Considering the severe complications associated with DNB, patients with elevated PLR should be seriously cared for in clinics.Keywords: diabetic autonomic neuropathy, neurogenic bladder, oxidative stress, inflammatory response

Qin Zhang,1,&ast; Fangming Shen,1,&ast; Qingfeng Wei,1,&ast; He Liu,2 Bo Li,1 Qian Zhang,3 Yueying Zhang3 1Xuzhou Medical University, Xuzhou City, Jiangsu Province, People’s Republic of China; 2Department of Anesthesiology, The Affiliated Huzhou Hospital, Zhejiang University School of Medicine; Huzhou Central Hospital, Huzhou City, Zhejiang Province, People’s Republic of China; 3Department of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou City, Jiangsu Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yueying ZhangDepartment of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou City, Jiangsu Province, People’s Republic of ChinaTel +86 138 1531 0789Email zyy0218@126.comPurpose: The aim of this study was to explore the associated risk factors of perioperative respiratory adverse events (PRAEs) in children undergoing airway surgery and establish and validate a nomogram prediction model for PRAEs.Patients and Methods: This study involved 709 children undergoing airway surgery between November 2020 and July 2021, aged ≤ 18 years in the affiliated hospital of Xuzhou Medical University. They were divided into training (70%; n = 496) and validation (30%; n = 213) cohorts. The least absolute shrinkage and selection operator (LASSO) was used to develop a risk nomogram model. Concordance index values, calibration plot, decision curve analysis, and the area under the curve (AUC) were examined.Results: PRAEs were found in 226 of 496 patients (45.6%) and 88 of 213 patients (41.3%) in the training and validation cohorts, respectively. The perioperative risk factors associated with PRAEs were age, obesity, degree of upper respiratory tract infection, premedication, and passive smoking. The risk nomogram model showed good discrimination power, and the AUC generated to predict survival in the training cohort was 0.760 (95% confidence interval, 0.695– 0.875). In the validation cohort, the AUC of survival predictions was 0.802 (95% confidence interval, 0.797– 0.895). Calibration plots and decision curve analysis showed good model performance in both datasets. The sensitivity and specificity of the risk nomogram model were calculated, and the result showed the sensitivity of 69.5% and 64.8% and specificity of 73.3% and 81.6% for the training and validation cohorts, respectively.Conclusion: The present study showed the proposed nomogram achieved an optimal prediction of PRAEs in patients undergoing airway surgery, which can provide a certain reference value for predicting the high-risk population of perioperative respiratory adverse events and can lead to reasonable preventive and treatment measures.Keywords: perioperative, adverse events, children, LASSO, nomogram

Shuting Yang,1 Qian Zhang,1 Yifan Xu,1 Futeng Chen,1 Fangming Shen,1 Qin Zhang,1 He Liu,2 Yueying Zhang3 1Xuzhou Medical University, Xuzhou City, Jiangsu Province, People’s Republic of China; 2Department of Anesthesiology, The Affiliated Huzhou Hospital, Zhejiang University School of Medicine; Huzhou Central Hospital, Huzhou City, Zhejiang Province, People’s Republic of China; 3Department of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou City, Jiangsu Province, People’s Republic of ChinaCorrespondence: Yueying ZhangDepartment of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou City, Jiangsu Province, People’s Republic of ChinaTel +86 138 1531 0789Email zyy0218@126.comPurpose: To develop a risk prediction nomogram of postoperative sleep disturbance (PSD) in patients undergoing non-cardiac surgery.Patients and methods: Data on 881 consecutive patients who underwent non-cardiac surgery at the Affiliated Hospital of Xuzhou Medical University between June 2020 and April 2021 were prospectively collected. Of these, we randomly divided 881 non-cardiac patients into two groups, training cohort (n = 617) and validation cohort (n = 264) at the ratio of 7:3. Characteristic variables were selected based on the data of training cohort through least absolute shrinkage and selection operator (LASSO) regression. Multivariate logistic regression was used to identify the independent risk factors associated with PSD that then were incorporated into the nomogram. The predictive performance of the nomogram was measured by concordance index (C index), receiver operating characteristic (ROC) curve, and calibration with 1000 bootstrap samples to decrease the over-fit bias.Results: PSD was found in 443 of 617 patients (71.8%) and 190 of 264 patients (72.0%) in the training and validation cohorts, respectively. The perioperative risk factors associated with PSD were female sex, anxiety, dissatisfaction of ward environment, absence of combined regional nerve block, postoperative nausea and vomiting (PONV), the longer duration stayed in post anesthesia care unit (PACU), the higher dose of midazolam and sufentanil, the higher postoperative numeric rating score for pain (NRS) score. Incorporating these 9 factors, the nomogram achieved good concordance indexes of 0.82 (95% confidence interval [CI], 0.78– 0.85) and 0.80 (95% CI, 0.74– 0.85) in predicting PSD in the training and validation cohorts, respectively, and obtained well-fitted calibration curves. The sensitivity and specificity (95% CIs) of the nomogram were calculated, resulting in sensitivity of 74.0% (70.0– 78.2%) and 75.3% (68.4– 81.7%) and specificity of 79.3% (72.5– 85.2%) and 70.3% (58.4– 80.7%) for the training and validation cohorts, respectively. Patients who had a nomogram score of less than 262 or 262 or greater were considered to have low or high risks of PSD presence, respectively.Conclusion: The proposed nomogram achieved an optimal prediction of PSD in patients undergoing non-cardiac surgery. The risks for an individual patient to harbor PSD can be determined by this model, which can lead to a reasonable preventive and treatment measures.Keywords: postoperative sleep disturbance, surgery, LASSO regression analysis, nomogram

Cong Zhou,1 Qiang Wu,1 Leqi He,1 Hui Zhang,1 Maosuo Xu,1 Baoyu Yuan,2 Zhi Jin,3 Fang Shen1 1Department of Clinical Laboratory Medicine, Shanghai Fifth People’s Hospital, Fudan University, Shanghai, People’s Republic of China; 2Department of Clinical Laboratory, Shanghai Children’s Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 3Department of Neurology, Shanghai Fifth People’s Hospital, Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Fang ShenDepartment of Clinical Laboratory Medicine, Shanghai Fifth People’s Hospital, Fudan University, No. 128, Ruili Road, Minhang District, Shanghai, 200240, People’s Republic of ChinaTel +86 18021073261Email shenfang5th@aliyun.comBackground: The convergence of carbapenem-resistance and hypervirulence in Klebsiella pneumoniae has led to a significant public health challenge. In recent years, there have been more and more reports on carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) isolates.Materials and Methods: Clinical data of patients infected with CR-hvKP from January 2019 to December 2020 in a tertiary hospital were retrospectively evaluated. The number of isolates of Klebsiella pneumoniae, hypermucoviscous Klebsiella pneumoniae (hmKP), carbapenem-resistant hypermucoviscous Klebsiella pneumoniae (CR-hmKP) and carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) collected during the period of 2 years was calculated. The antimicrobial resistance gene, virulence-associated gene, capsular serotype gene and multilocus sequence typing (MLST) of CR-hvKP isolates were detected by PCR.Results: During the study period, a total of 1081 isolates of non-repeat Klebsiella pneumoniae were isolated, including 392 isolates of hypermucoviscous Klebsiella pneumoniae (36.3%), 39 isolates of CR-hmKP (3.6%), and 16 isolates of CR-hvKP (1.5%). About 31.2% (5/16) of CR-hvKP were isolated from 2019, and 68.8% (11/16) of CR-hvKP were isolated from 2020. Among the 16 isolates of CR-hvKP, 13 isolates were ST11 and serotype K64, 1 isolate was ST11 and serotype K47, 1 isolate was ST23 and serotype K1, and 1 isolate was ST86 and serotype K2. The virulence-associated genes entB, fimH, rmpA2, iutA, iucA were present in all of 16 CR-hvKP isolates, followed by mrkD (n=14), rmpA (n=13), aerobactin (n=2), allS (n=1). Sixteen CR-hvKP isolates all carry carbapenemase gene blaKPC-2 and extended-spectrum β-lactamase gene blaSHV. ERIC-PCR DNA fingerprinting results showed that 16 CR-hvKP isolates were highly polymorphic, and there were significant differences in bands among the isolates, presenting a sporadic state.Conclusion: Although CR-hvKP was sporadically distributed, it showed an increasing trend year by year. Therefore, clinical attention should be paid, and necessary measures should be taken to avoid the cloning and transmission of superbacterium CR-hvKP.Keywords: Klebsiella pneumoniae, carbapenem-resistant, hypervirulent, hypermucoviscous, epidemiology

Lei Liang,1,2,* Ming-Da Wang,3,* Yao-Ming Zhang,4 Wan-Guang Zhang,5 Cheng-Wu Zhang,1 Wan Yee Lau,3,6 Feng Shen,3 Timothy M Pawlik,7 Dong-Sheng Huang,1,2,8 Tian Yang1– 3,8 1Department of Hepatobiliary, Pancreatic and Minimal Invasive Surgery, Zhejiang Provincial People’s Hospital, People’s Hospital of Hangzhou Medical College, Zhejiang, People’s Republic of China; 2School of Clinical Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, People’s Republic of China; 3Department of Hepatobiliary Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University (Navy Medical University), Shanghai, People’s Republic of China; 4The 2nd Department of Hepatobiliary Surgery, Meizhou People’s Hospital, Guangdong, People’s Republic of China; 5Department of Hepatic Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 6Faculty of Medicine, The Chinese University of Hong Kong, New Territories, Hong Kong, China; 7Department of Surgery, Ohio State University, Wexner Medical Center, Columbus, OH, USA; 8The Key Laboratory of Tumor Molecular Diagnosis and Individualized Medicine of Zhejiang Province, Zhejiang Provincial People’ s Hospital (People’ s Hospital of Hangzhou Medical College), Hangzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Tian YangDepartment of Hepatobiliary, Pancreatic and Minimal Invasive Surgery, Zhejiang Provincial People’s Hospital, People’s Hospital of Hangzhou Medical College, No. 158, Shangtang Road, Hangzhou, Zhejiang, 310014, People’s Republic of ChinaEmail yangtiandfgd@hotmail.comDong-Sheng HuangDepartment of Hepatobiliary, Pancreatic and Minimal Invasive Surgery, Zhejiang Provincial People’s Hospital, People’s Hospital of Hangzhou Medical College, No. 158, Shangtang Road, Hangzhou, Zhejiang, 310014, People’s Republic of ChinaEmail Huangdongshengsj@hotmail.comBackground: High alpha-fetoprotein (AFP) expressions (> 400 ng/mL) are associated with poor oncological characteristics for hepatocellular carcinoma (HCC). However, prognosis after liver resection for high-AFP HCC is poorly studied. To investigate long-term recurrence and survival after hepatectomy for high-AFP HCC, and to identify the predictive value of postoperative incomplete biomarker response (IBR) on overall survival (OS) and recurrence-free survival (RFS).Methods: Patients undergoing curative resection for high-AFP HCC were analyzed. According to the decline magnitude of serum AFP as measured at first follow-up (4∼ 6 weeks after surgery), all patients were divided into the complete biomarker response (CBR) and IBR groups. Characteristics, recurrence, and survival rates were compared. Univariate and Multivariate Cox-regression analyses were performed to identify independent predictors associated with poorer OS and RFS after liver resection for high-AFP HCC.Results: Among 549 patients, the overall and early recurrence rates in patients with IBR were significantly higher than patients with CBR (97.8%vs.56.4%, and 92.5%vs.33.3%, both P< 0.001). On multivariate analysis, postoperative IBR was the strongest risk factor with the highest hazard ratio in predicting poor OS (HR 2.97; 95% CI 2.49∼ 3.45; P< 0.001) and RFS (HR 4.29; 95% CI 3.31∼ 5.55; P< 0.001).Conclusion: Postoperative biomarker response of serum AFP can be used in predicting recurrence and survival for high-AFP HCC patients. Once postoperative IBR was identified at first follow-up, subsequent enhanced recurrence surveillance and available treatments against recurrence should actively be considered.Keywords: hepatocellular carcinoma, alpha-fetoprotein, hepatectomy, survival, recurrence

Wenwen Chen,1,* Kun Chen,2,* Zhixiao Xu,3 Yepeng Hu,1 Yiying Liu,1 Wenyue Liu,1 Xiang Hu,1 Tingting Ye,1 Jing Hong,1 Hong Zhu,1 Feixia Shen1 1Department of Endocrinology and Metabolism, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 2Department of Thoracic Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 3Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Feixia ShenDepartment of Endocrinology and Metabolism, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of ChinaEmail shenfeixia@wmu.edu.cnPurpose: Elevated platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) are associated with poor outcomes in various diseases. The objectives of this study were to explore the utility of PLR and NLR in predicting all-cause mortality in patients with diabetic foot ulcers (DFU) undergoing amputations.Patients and Methods: A retrospective observational study was performed that included a total of 348 DFU patients undergoing amputations. The primary end-point was all-cause death. According to the PLR and NLR cut-off values, patients were divided into two groups and Kaplan–Meier survival curves were constructed. Multivariable Cox regression was conducted to test the independent predictors of mortality in the study cohort.Results: All-cause mortality was significantly higher in patients with a high PLR/NLR compared to those with a low PLR/NLR. In the low NLR group, the overall survival (OS) rates at 1, 3, and 5 years after amputation were 96.8%, 84% and 80.1%, respectively (p=0.001). In the high NLR group the corresponding OS rates at 1, 3, and 5 years were 85.2%, 58.6% and 23.9% (p< 0.001). According to the multivariate analysis, age (HR 1.074, 95% CI 1.045– 1.104, p< 0.001), Wagner classification (HR 2.274, 95% CI 1.351– 3.828, p=0.002), PLR (HR 1.794, 95% CI 1.014– 3.174, p=0.045), NLR (HR 2.029, 95% CI 1.177– 3.499, p=0.011), creatinine (HR 1.003, 95% CI 1.001– 1.004, p< 0.001) and direct bilirubin (HR 1.154, 95% CI 1.081– 1.232, p< 0.001) were independent predictors of mortality following amputation.Conclusion: Postoperative PLR and NLR values may be reliable predictive biomarkers of mortality in patients following amputation for DFU. Considering the high mortality in those patients, the patients with elevated PLR/NLR should be given more intensive in clinical practice.Keywords: amputation, platelet-to-lymphocyte ratio, PLR, neutrophil-to-lymphocyte ratio, NLR, diabetic foot ulcer, mortality

Xinyi Zhu,1,2,* Mengqiao Xu,3,* Xingpeng Zhao,4,* Fei Shen,1,2 Changgeng Ruan,1,2 Yiming Zhao1,2 1Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of the Ministry of Health, The First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu, People’s Republic of China; 2Collaborative Innovation Center of Hematology, Soochow University, Suzhou 215006, Jiangsu, People’s Republic of China; 3Department of Laboratory Medicine, The Affiliated Municipal Hospital of Taizhou University, Taizhou 318000, Zhejiang, People’s Republic of China; 4Clinical Laboratory Center, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang 471000, Henan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yiming ZhaoJiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of the Ministry of Health, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou 215006, Jiangsu, People’s Republic of ChinaTel + 86-512-67781379Fax + 86-512-65113556Email zhaoyimingbox@163.comBackground: Podoplanin (PDPN) is a type-1 membrane sialoglycoprotein that is expressed in many cancer tumors including breast cancer; nonetheless, its roles in tumor occurrence, development, and metastasis are unclear. In this study, we aimed to investigate the clinical significance of plasma soluble PDPN (sPDPN) levels in patients with breast cancer and its significance in the diagnosis and metastasis.Materials and Methods: Blood samples from healthy controls (CTL), patients with fibroadenomas of breast (FOB), and breast cancer (pathological type: invasive ductal carcinoma, IDC) were collected. sPDPN levels in the plasma of CTL and patients with FOB and IDC were measured by the ELISA.Results: The plasma sPDPN levels in IDC patients (159 cases, 22.59± 3.70 ng/mL) were higher than those in FOB patients (50 cases, 8.29± 1.09 ng/mL; P< 0.05) and CTL (100 cases, 1.21± 0.12 ng/mL; P< 0.0001). The sPDPN levels in patients at stage III and stage IV (30.08± 4.66 ng/mL) were higher than in patients at stage I and stage II (11.84± 1.12 ng/mL; P=0.005). The sPDPN levels in patients with high-moderate and moderate differentiation (17.50± 3.02 ng/mL) were lower than those in patients with moderately low and low differentiation (35.73± 4.26 ng/mL; P=0.026). The sPDPN levels in patients with metastasis (30.60± 4.27 ng/mL) were much higher than those in patients without metastasis (13.02± 1.30 ng/mL; P=0.017).Conclusion: Plasma sPDPN may be used as a new marker for the determination of the clinical stage, differentiation degree, and metastasis status of breast cancer.Keywords: podoplanin, breast cancer, ELISA, monoclonal antibody

Qiang Zhang,1,2,* Fei Shen,1,2,* WenQing Shen,1,2 Jie Xia,1,2 Jing Wang,1,2 Yu Zhao,1,3 Zhe Zhang,1,2 Yi Sun,1,2 Min Qian,4 ShuZhe Ding1,2 1Key Laboratory of Adolescent Health Assessment and Exercise Intervention of Ministry of Education, East China Normal University, Shanghai 200241, People’s Republic of China; 2School of Physical Education & Health, East China Normal University, Shanghai 200241, People’s Republic of China; 3Center for Physical Education, Xi’an Jiaotong University, Xi’an, Shaanxi 710049, People’s Republic of China; 4Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, People’s Republic of China*These authors contributed equally to this workCorrespondence: ShuZhe Ding Tel +86 13601798505Email szding@tyxx.ecnu.edu.cnObjective: Ketogenic diet (KD) and high-intensity interval training (HIIT) have preclinical benefits for type 2 diabetes (Db). However, the health risks of long-term KD use in diabetes should be ascertained and prevented. We hypothesized that KD-induced liver fibrosis in type 2 diabetic mice could be ameliorated by HIIT.Methods: Streptozotocin-induced type 2 diabetic mice were divided into high-fat diet (HFD) control (Db+HFD+Sed), KD control (Db+KD+Sed), HFD coupled with HIIT (Db+HFD+HIIT), and KD coupled with HIIT (Db+KD+HIIT) groups (n=6, per group). Control mice were kept in sedentary (Sed), while HIIT group mice underwent 40-minute high-intensity interval training three alternate days per week. After 8-week intervention, the indicators of body weight and insulin resistance, oxidative stress markers, hepatic fibrosis, genetic and protein expression of related pathways were tested.Results: We found that fasting blood glucose level was reduced in the Db+HFD+HIIT, Db+KD+Sed, and Db+KD+HIIT groups. Insulin sensitivity was increased in diabetic mice of these groups, whereas ROS levels were decreased in mice that underwent HIIT. The immunohistochemical staining of liver, serum index, and hepatic parameters of diabetic mice in the KD group revealed liver fibrosis, which was significantly attenuated by HIIT. Besides, these effects of HIIT were the outcome of hepatic stellate cell’s inactivation, reduced protein expression of matrix metalloproteinases and tissue inhibitor of metalloproteinases, and the inhibition of TGF-β 1/Smad signaling.Conclusion: KD had a profound fibrotic effect on the liver of type 2 diabetic mice, whereas HIIT ameliorated this effect. KD did not show any apparent benefit as far as glucose tolerance and homeostasis were concerned. Concisely, our results demonstrated that KD should be coupled with HIIT for the prevention and preclinical mitigation of type 2 diabetes.Keywords: diabetes, ketogenic diet, high-intensity interval training, hepatic fibrosis, ROS, TGF-β 1/Smad signal

Chenlu Zhang,1 Zhiming Wang,1 Rongyuan Zhuang,1 Xi Guo,1 Yi Feng,1 Feng Shen,1 Wenshuai Liu,2 Yong Zhang,3 Hanxing Tong,3 Wending Sun,4 Jun Liu,4 Guan Wang,4 Chun Dai,4 Weiqi Lu,3 Yuhong Zhou1 1Department of Medical Oncology, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China; 2Department of General Surgery, Shanghai Public Health Clinical Center, Shanghai, People’s Republic of China; 3Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China; 4GenomiCare Biotechnology Co. Ltd, Shanghai, People’s Republic of ChinaCorrespondence: Weiqi LuDepartment of General Surgery, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, People’s Republic of ChinaEmail lu.weiqi@zs-hospital.sh.cnYuhong ZhouDepartment of Medical Oncology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, People’s Republic of ChinaEmail zhou.yuhong@zs-hospital.sh.cnAbstract: We report two inflammatory myofibroblastic tumor (IMT) patients with ALK fusions (RRBP-ALK and TNS1-ALK, respectively). They both received tumor resection surgery and treatment with ALK inhibitors crizotinib followed by alectinib, and upon receiving each of the drugs, showed a brief response, then experienced recurrence or progression of the disease. During the treatment, whole exome sequencing (WES) and RNA sequencing (RNA-Seq) were applied to monitor potential drug-induced gene mutation and expression changes. A novel, secondary mutation in ALK exon 23 (L1196Q) was identified in patient 1 after alectinib resistance developed. Guided by this result, a newer ALK inhibitor, ceritinib was prescribed. The patient was able to achieve a partial response (PR) and is in good condition as of the manuscript date. On the contrary, there was no secondary mutation identified in ALK in patient 2 after drug resistance. While the expression of PTCH1, a negative regulator of the sonic hedgehog (SHH) signaling pathway, was significantly reduced at the time after the treatment with crizotinib before that of alectinib. The expression of PTCH1 was also reduced after the treatment with alectinib. It was reported that ALK can exert its biological functions partially by activating SHH signaling pathway. The down-regulation of PTCH1 suggests the compensatory activation of SHH pathway may cause resistance to ALK inhibitors in IMT. Going forward, monitoring gene mutation and expression changes through DNA and RNA sequencing will be able to offer opportunities to investigate potential mechanisms of drug resistance and will help to achieve precise prescription for better treatment outcomes.Keywords: inflammatory myofibroblastic tumor, crizotinib resistance, alectinib resistance, ALK L1196Q, sonic hedgehog pathway

Rong Luo ,* Xiaochen Sun ,* Feiyan Shen, Bin Hong, Zilong Wang Department of Cardiology, Qingpu Medical Center, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zilong Wang Email wangthinker123@outlook.comObjective: To investigate the effects of high-dose rosuvastatin on ventricular remodelling and cardiac function in ST-segment elevation myocardial infarction (STEMI).Materials and Methods: From January 2017 to March 2019, the clinical data of 93 patients with STEMI were collected and analysed, with 46 cases in the conventional-dose group (rosuvastatin, 10 mg/d) and 47 cases in the high-dose group (rosuvastatin, 20 mg/d). Blood lipid (TC, TG, LDL-C and HDL-C), serum inflammatory markers (hs-CRP, IL-6, TNF-α and ICAM-1), ventricular remodelling markers (NT-pro BNP, MMP-9, TIMP-4 and Gal-3) and indicators of cardiac function (LVESD, LVESD, LVESV, LVEDV, IVST and LVEF) were collected from all patients at the time of admission and 8 weeks after rosuvastatin treatment.Results: After treatment with rosuvastatin for 8 weeks, compared with those in conventional-dose group, the levels of TC, TG, LDL-C, hs-CRP, IL-6, TNF-α, ICAM-1, NT-pro BNP, MMP-9 and Gal-3 in the high-dose group decreased significantly (P< 0.05), while the increase of HDL-C and TIMP-4 levels was more obvious (P< 0.05) than that in the conventional-dose group. Moreover, LVEF was significantly higher (P< 0.05) and LVESD, LVESD, LVESV, LVEDV and IVST were significantly lower (P< 0.05) after treatment than before treatment in both groups. The improvement of cardiac ultrasound results in the high-dose group was more significant than that in the conventional-dose group (P< 0.05).Conclusion: This study suggests that high-dose rosuvastatin was better than conventional-dose rosuvastatin for improving blood lipid metabolism, reducing the inflammatory response, and preventing and treating ventricular remodelling and myocardial fibrosis, indicating that high-dose rosuvastatin had stronger therapeutic effect on STEMI than conventional-dose rosuvastatin.Keywords: rosuvastatin, ST elevation myocardial infarction, blood lipids, inflammatory factor, ventricular remodelling, myocardial fibrosis

Zheng-Liang Chen,1,* Cheng-Wu Zhang,2,* Lei Liang,2,* Han Wu,3,* Wan-Guang Zhang,4 Yong-Yi Zeng,5 Wei-Min Gu,6 Ting-Hao Chen,7 Jie Li,8 Yao-Ming Zhang,9 Hong Wang,10 Ya-Hao Zhou,11 Chao Li,2 Yong-Kang Diao,2 Wan Yee Lau,3,12 Meng-Chao Wu,3 Feng Shen,3 Tian Yang,3 Ying-Jian Liang1 1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Harbin Medical University, Heilongjiang, People’s Republic of China; 2Department of Hepatobiliary, Pancreatic and Minimal Invasive Surgery, Zhejiang Provincial People’s Hospital, People’s Hospital of Hangzhou Medical College, Zhejiang, People’s Republic of China; 3Department of Hepatobiliary Surgery, Eastern Hepatobiliary Surgery Hospital (Navy Medical University), Second Military Medical University, Shanghai, People’s Republic of China; 4Department of Hepatic Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 5Department of Hepatobiliary Surgery, Mengchao Hepatobiliary Hospital, Fujian Medical University, Fujian, People’s Republic of China; 6The First Department of General Surgery, The Fourth Hospital of Harbin, Heilongjiang, People’s Republic of China; 7Department of General Surgery, Ziyang First People’s Hospital, Sichuan, People’s Republic of China; 8Department of Hepatobiliary Surgery, Fuyang People’s Hospital, Anhui, People’s Republic of China; 9The 2nd Department of Hepatobiliary Surgery, Meizhou People’s Hospital, Guangdong, People’s Republic of China; 10Department of General Surgery, Liuyang People’s Hospital, Hunan, People’s Republic of China; 11Department of Hepatobiliary Surgery, Pu’er People’s Hospital, Yunnan, People’s Republic of China; 12Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, SAR, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ying-Jian LiangDepartment of Hepatobiliary Surgery, The First Affiliated Hospital of Harbin Medical University, No. 23, Youzheng Road, Heilongjiang 150006, People’s Republic of ChinaEmail genomeliang@hotmail.comTian YangDepartment of Hepatobiliary Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University (Navy Medical University), No. 225, Changhai Road, Shanghai 200438, People’s Republic of ChinaEmail yangtiandfgd@hotmail.comBackground: With an increase in life expectancy and improvement of surgical safety, more elderly patients with hepatocellular carcinoma (HCC), even with large tumors, are now considered for hepatectomy. This study aimed to clarify the impact of age on short- and long-term outcomes after major hepatectomy (≥ 3 segments) for large HCC (≥ 5 cm).Patients and Methods: Using a multicenter database, patients who underwent curative-intent major hepatectomy for large HCC between 2006 and 2016 were identified. Postoperative morbidity and mortality, overall survival (OS) and recurrence-free survival (RFS) were compared between the elderly (≥ 65 years) and younger (< 65 years) patients. Univariable and multivariable Cox-regression analyses were performed to identify the risk factors of OS and RFS in the entire and elderly cohorts, respectively.Results: Of 830 patients, 92 (11.1%) and 738 (88.9%) were elderly and younger patients, respectively. There were no significant differences in postoperative 30-day mortality and morbidity between the two groups (5.4% vs 2.6% and 43.5% vs 38.3%, both P> 0.05). The 5-year OS and RFS rates in elderly patients were also comparable to younger patients (35.0% vs 33.2% and 20.0% vs 20.8%, both P> 0.05). In the entire cohort, multivariable Cox-regression analyses identified that old age was not independently associated with OS and RFS. However, in the elderly cohort, preoperative alpha-fetoprotein level > 400 μg/L, multiple tumors, macrovascular invasion and microvascular invasion were independently associated with decreased OS and RFS.Conclusion: Carefully selected elderly patients benefited from major hepatectomy for large HCC as much as younger patients, and their long-term prognosis was determined by preoperative alpha-fetoprotein level, tumor number and presence of macro- or micro-vascular invasion.Keywords: hepatocellular carcinoma, hepatectomy, survival, recurrence, elderly

Feng Shen,1,* Xiangou Pan,2,* Min Li,3,* Yixing Chen,2 Ying Jiang,3 Jian He2 1Department of Medical Oncology, Zhongshan Hospital, Fudan University, Shanghai 200032, People’s Republic of China; 2Department of Radiation Oncology, Zhongshan Hospital, Fudan University, Shanghai 200032, People’s Republic of China; 3Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jian HeDepartment of Radiation Oncology, Zhongshan Hospital, Fudan University, Shanghai 200032, People’s Republic of ChinaTel/ Fax +86-021- 64041990Email hejian62@163.comYing JiangDepartment of General Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, People’s Republic of ChinaTel/Fax +86-21-64041990Email jiang.ying3@zs-hospital.sh.cnBackground: Breast cancer remains a great threat to females worldwide. As a recently defined programmed cell death pathway that associates with immune activation, RIP1/RIP3/MLKL necroptosis signaling has been implicated in a variety of diseases. The present study aimed to investigate the role of RIP1/RIP3/MLKL signaling in breast cancer cell proliferation and metastasis in vivo and in vitro.Methods: Western blot and quantitative real-time PCR were performed to evaluate the activation of necroptosis signaling in clinical human breast cancer tissues. Correlation of necroptosis signaling markers with clinicopathological parameters was statistically assessed. Cell viability assay, colony formation assay, wound healing assay, and transwell migration and invasion assays were performed to investigate the effects of necroptosis inhibition on breast cancer cell proliferation and metastasis.Results: Clinical breast cancer tissues showed significantly higher levels of tumor necrosis factor alpha (TNFα), RIP1, RIP3 and MLKL at both mRNA and protein levels as compared with their paired non-cancerous tissues. Phosphorylation of RIP3 and MLKL was also remarkably provoked. Statistics showed that both RIP1 and MLKL positively correlated with cancer parameters such as N-cadherin (p=0.002 for RIP1 and p=0.021 for MLKL) and Ki67 (p=0.031 for RIP1 and p=0.05 for MLKL). The MLKL expression level significantly correlated with tumor size (p=0.001) and the proliferation indicator Ki67 (p=0.018). In addition, pharmacological inhibition of the necroptosis signaling using necrostatin-1 promoted breast cancer cell proliferation and colony formation by approximately 50%. Blockade of necroptosis signaling also accelerated wound healing process and cell transmigration in breast cancer cells.Conclusion: Our results suggested that pharmacological inhibition of necroptosis promoted breast cancer cell proliferation and metastasis. Modulation of tumor cell necroptosis might represent a novel strategy as to breast cancer treatment.Keywords: breast cancer, necroptosis, RIP1, MLKL, proliferation, metastasis

As Australia is the main iron ore importing country for China, the abundant mineral resources of Australia are the primary raw materials for the sintering process. To better understand the properties of different iron ores from Australia and then guide the sintering process, this study selected four types of Australian ore and one type of domestic ore, and their properties were investigated under different conditions. The experiment of single iron ore sintering was studied to examine the influence of different iron ores on the metallurgical properties of sinter. From this study, the following results were obtained: GG showed poor fluidity and higher assimilability temperature, but the bonding phase strength was the highest; YD showed better fluidity and lower assimilability temperature, whereas SJY (domestic ore) showed better fluidity and higher bonding phase strength, and lower assimilability temperature. The influence mechanism of iron ore on the fluidity was further analyzed by using SEM and ion theory of slag. With the increase SiO2 content of iron ore, the fluidity index increased, the main reason was that the amount of liquid phase increased and the melting point decreased during the sintering. However, an excessive amount of SiO2 caused the decrease of fluidity index of iron ore; the main reason was that the fluidity of the liquid phase itself decreased and secondary hematite appeared. In the case of the higher SiO2 content of iron ore, the main bonding phase was calcium silicate. With the decrease of SiO2 content, the calcium silicate transformed into calcium ferrite. The main reason for this was that the Gibbs free energy of calcium ferrite and dicalcium ferrite (2CaO·SiO2) was higher than that of calcium silicate in the temperature ranges of 400-1600 K. The reduction degree of YD was the highest in all the cases and that of GG was the lowest. Activation energies of 5.39, 3.14, 3.51, 4.47 and 2.92 kJ/mol were obtained for the reduction of GG, PB, BH, SJY, and YD, respectively. In all the cases, the reaction corresponded to the model function of F1(α), and the integral form was -ln(1-α)=kt. Through this investigation, it could be concluded that the most appropriate ore category for sinter pot was YD.

He-Qi Bu, Feng Shen, Junhui Cui Department of Coloproctological Surgery, Tongde Hospital of Zhejiang Province, Hangzhou 310012, People’s Republic of ChinaCorrespondence: Junhui CuiDepartment of Coloproctological Surgery, Tongde Hospital of Zhejiang Province, 234 Gu-Cui Road, Hangzhou 310012, People’s Republic of ChinaTel/Fax +86 571 8997 2373Email 13958105761@163.comBackground: Oridonin, the main active component of Rabdosia rubescens, has been demonstrated to have anti-tumor effect on all kinds of cancer cells through various mechanisms and it has shown antitumor activity in some tumors partially via the suppression of TGF-β/Smads signaling pathway. The aim of this study was to explore the anticancer effect of oridonin on human colon carcinoma and underlying mechanism in vitro and vivo.Methods: CCK-8 assay was employed to assess cell viability. The key target genes and proteins involved in TGF-β/Smads pathway was detected by RT-PCR, Western blotting and immunohistochemistry. The orthotopic transplantation tumor model of colon cance LOVO cell was introduced to detect anti-cancer effects in vivo.Results: Oridonin inhibited the proliferation of colon cancer LOVO cells in a concentration and time dependent manner. In addition, oridonin reduced the levels of Smad2, Smad3, Smad4, PAI-1 and the phosphorylation of Smad2 and Smad3 induced by TGF-β1 in vitro. Subsequently, we established an orthotopically implanted tumor model in nude mice and found that oridonin treatment significantly suppressed tumor growth, and which was accompanied by the down-regulation of Smad2, Smad3, Smad4, PAI-1 and p-Smad2, p-Smad3 expression levels.Conclusion: Our present study demonstrated that the growth inhibition of colon cancer by oridonin could be partially mediated through discontinuing TGF-β1/Smads-PAI-1 signaling pathway, suggesting it as a promising agent in treating colorectal cancer.Keywords: oridonin, colon cancer, TGF-β/Smads pathway, PAI-1

Fengru Zhou,1 Feng Shen,1 Zhuojun Zheng,2 Jincheng Ruan11Department of Anus and Intestine Surgery, The Tongde Hospital of Zhejiang Province, Hangzhou, Zhejiang Province, People’s Republic of China; 2Department of Hematology, The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu Province, People’s Republic of ChinaBackground: Colorectal cancer is a heterogeneous disease with complex genetic and epigenetic changes. LncRNA has recently been regarded as the biomarker in cancers. Novel biomarkers in colon cancer need to be identified.Purpose: The objective of this study was to identify the differentially expressed lncRNAs between colon cancer tissue and adjacent tissue, as well as to explore its biological functions.Patients and methods: There were 130 colon cancer patients included in this study. Of them, 6 colon cancer samples and 3 normal samples were selected for microarray profiling. Another 121 colon cancer samples with complete clinical information were used for immunohistochemical assay and survival analysis. Microarray analysis was performed to determine the differentially expressed lncRNAs between colon cancer tissue and adjacent tissue. Gain-of-function experiments was conducted in vitro and in vivo. In situ hybridization and survival analysis were applied to determine the prognostic impact on survival.Results: LncRNA XIRP2-AS1 was significantly less expressed in colon cancer tissue. XIRP2-AS1 was remarkably downregulated in colon cancer tissues and cell lines. Functionally, XIRP2-AS1 could inhibit the proliferation and invasion ability of colon cancer cells in vitro and in vivo. Clinical sample analysis showed that XIRP2-AS1 had a favorable impact on the overall survival and progression free survival of patients with colon cancer. miR-182 was validated as the target of XIRP2-AS1 according to luciferase reporter assays, RNA immunoprecipitation and RNA pull down.Conclusions: Our results suggested that XIRP2-AS1 may act as a favorable biomarker for patients with colon cancer.Keywords: colon cancer, XIRP2-AS1, proliferation, invasion, survival

Han-Song Sheng,1 Fang Shen,2 Nu Zhang,1 Li-Sheng Yu,1 Xiang-Qi Lu,1 Zhe Zhang,1,3 Huang-Yi Fang,1,3 Ling-Li Zhou,4 Jian Lin11Department of Neurosurgery, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 2Department of Orthopedic Surgery‘s Spine Division, The Affiliated Hospital of Medical School of Ningbo University, Ningbo, People’s Republic of China; 3School of the 2nd Clinical Medical Sciences, Wenzhou Medical University, Wenzhou, People’s Republic of China; 4Department of Pathology, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of ChinaBackground: Although meningiomas are common intracranial tumors, multiple meningiomas (MMs) are rare entities in patients without neurofibromatosis type 2. Previous studies suggest most sporadic MMs are of monoclone in origin.Objective: To elucidate the clonal relationship between two sporadic meningiomas from the same patient by using the next-generation sequencing (NGS) platform.Methods: Two MMs, located frontally and parietally on the right side, were surgically removed from a 52-year-old male. Pathological examinations and whole exome sequencing were performed on tumor samples, followed by Sanger sequencing validation.Results: MMs were diagnosed as secretory and fibrous subtypes, respectively, on histology (WHO grade I) and tumor DNA exhibited distinctive somatic mutation patterns. Specifically, the secretory subtype carried more single nucleotide variant while the fibrous subtype had much higher copy number variation. Besides, the two tumors demonstrated different mutation profiles in predisposing genes and known driver mutations. For example, the secretory subtype had missense mutations in TRAF7 and KLF4, while the fibrous subtype had frameshift deletion of NF2 gene in addition to copy number loss of NF2 and SMARCB1, genetic events that have already been associated with the development of meningiomas. Significantly mutated gene analysis revealed novel mutations of LOC729159 in the secretory subtype and RPGRIP1L and DPP6 in the fibrous subtype. Sanger sequencing validated important point mutations in TRAF7 (c.1678G>A, p.G560S), KLF4 (c.1225A>C, p.K409Q) and CDH11 (c.169T>G, p.W57G).Conclusion: Our data suggest the two meningiomas might develop independently in this patient and molecular subtyping by NGS is a valuable supplement to conventional pathology. Further study is needed to ascertain whether these novel genetic events are tumorigenic or simply passenger mutations, as well as their clinical implicationsKeywords: multiple meningiomas, whole exome sequencing, secretory meningioma, fibrous meningioma, TRAF7, NF2

Feifei Shen,1,* Xin Dong,1,* Xin Zhou,2 Lanyun Yan,1 Qi Wan11Department of Neurology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, People’s Republic of China; 2Department of Ophthalmology, The Affiliated Hospital of Nanjing University of Traditional Chinese Medicine, Nanjing, Jiangsu 210029, People’s Republic of China *These authors contributed equally to this workBackground and purpose: It has been generally thought that activation and sensitization of the trigeminovascular system may contribute to the pathogenesis of migraine. Nevertheless, there is little evidence on abnormalities in peripheral trigeminal afferent nerves from humans in vivo. Alterations of corneal nerves from the ophthalmic branch of the trigeminal nerve may support the notion that trigeminal afferent nerves are involved in migraine pathophysiology. The aim of the present study was to investigate the structural changes in corneal subbasal nerve plexus in patients with episodic migraine (EM) with in vivo confocal microscope (IVCM).Methods: In this cross-sectional observational study, 10 EM patients and 10 age- and sex-matched healthy controls were included. Analysis of IVCM images with Image J software was performed to quantify the changes in the corneal subbasal nerve plexus.Results: EM patients showed an increase in nerve fiber length (25.0±2.65 vs 22.3±2.41 mm/mm,2 p=0.047) and nerve fiber density (36.3±7.29 vs 30.5±6.19 fibers/mm,2 p=0.104) as compared with normal controls, but this difference was not statistically significant. Nerve branching and tortuosity were significantly increased in the EM subjects compared to the normal subjects (91.3±13.8 vs 75.0±14.2 branches/mm,2 p=0.030 and 2.30±0.46 versus 1.63±0.52, p=0.011, respectively). In addition, nerve sprouts and increased number of Langerhans cells were observed in the EM patients.Conclusion: The morphologic changes of corneal subbasal nerve plexus and Langerhans cell aggregation suggest the presence of nerve regeneration and inflammation in EM. Furthermore, the alterations of corneal nerves from the ophthalmic branch of the trigeminal nerve offer support for the hypothesis that the peripheral trigeminal system may be involved in the pathogenesis of migraine.Keywords: migraine, corneal nerves, subbasal nerve plexus, in vivo confocal microscopy

Peng Zheng,1,* Xiaoqian Wang,2,* Zhong Hong,1 Feixia Shen,2 Qiyu Zhang3 1Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China; 2Department of Endocrinology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China; 3Department of Hepatobiliary Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China *These authors contributed equally to this work Background: Preoperative high blood glucose levels are closely associated with poor performance and high mortality in cancer patients. This study was designed to investigate the relationship between preoperative fasting hyperglycemia and the prognosis of patients with gallbladder cancer (GBC) after undergoing GBC radical surgery.Patients and methods: A retrospective analysis of 83 eligible patients who underwent GBC radical surgery between 2007 and 2016 was performed. Factors affecting overall survival (OS) and recurrence-free survival (RFS) were analyzed by univariate and multivariate analyses.Results: Of the 83 patients, 35 (42.2%) had preoperative fasting hyperglycemia. The median OS of the enrolled patients was 12 months. The median OS in patients with fasting hyperglycemia before surgery was 18 months, which was shorter than for patients with normal fasting blood glucose levels before surgery (47 months, P

Yi-jun Lyu,1,2,* Fang Shen,3,* Yun Yan,4,5 Ming-zhu Situ,4,5 Wei-zhu Wu,1,2 Guo-qiang Jiang,3 Ya-ya Chen4,5 1Department of Thyroid and Breast Surgery, Ningbo Medical Center Lihuili Eastern Hospital, Ningbo 315040, Zhejiang, China; 2Department of Thyroid and Breast Surgery, Taipei Medical University Ningbo Medical Center, Ningbo 315040, Zhejiang, China; 3Department of Orthopaedic Surgery’s Spine Division, The Affiliated Hospital of Medical School of Ningbo University, Ningbo 315020, Zhejiang, China; 4Department of Ultrasound, Ningbo Medical Center Lihuili Eastern Hospital, Ningbo 315040, Zhejiang, China; 5Department of Ultrasound, Taipei Medical University Ningbo Medical Center, Ningbo 315040, Zhejiang, China *These authors contributed equally to this work Objective: Ultrasound-guided fine-needle aspiration biopsy (US-FNAB) is a safe and effective method of screening malignant thyroid nodules such as papillary thyroid carcinoma. However, not much data are available regarding the diagnostic efficacy of US-FNAB for papillary thyroid microcarcinoma (≤10 mm in diameter). We aim to compare the diagnostic efficacy of US-FNAB on thyroid nodules between two groups divided by a diameter of 10 mm by correlating the cytological results of US-FNAB with the histopathologic diagnoses in selected patients. Patients and methods: Eight hundred twenty-two thyroid nodules (Group A: diameter ≤10 mm, n=620; Group B: diameter >10 mm, n=202) from 797 patients treated between March 2014 and June 2017 were retrospectively evaluated. Only nodules with Thyroid Imaging Reporting and Data System (TIRADS) categories 4–6 were enrolled and sampled by US-FNAB, followed by surgical resection. Results: According to The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) diagnostic categories, 94 thyroid nodules were classified as I, III and IV, and were excluded from the analysis. The resultant 728 thyroid nodules from 721 patients were analyzed. The malignant tendency (TBSRTC V and VI) rates on US-FNAB were 88.2% and 84.6% (P=0.202) in Group A and Group B, respectively, and the malignant rates were 89.5% and 86.9% (P=0.330), respectively, on histopathology. There was a high concordance between cytology and histopathology diagnoses (kappa value =0.797), and no statistical difference in terms of US-FNAB accuracy was found between the two groups (P=0.533). Conclusion: For thyroid nodules of TIRADS category 4–6, the diagnostic efficacy of US-FNAB is similar for thyroid nodules either smaller or greater than 10 mm in their maximum diameter. Keywords: ultrasound-guided fine-needle aspiration biopsy, thyroid nodules, microcarcinoma, histopathology, cytopathology

The edge and internal morphology of coke with different contents of UPC (unburned pulverized coal) after reaction with CO2 were analyzed by SEM. The influence of UPC on CRI (coke reactivity)，CSR (coke strength after reaction) and apparent porosity was also studied. The synthetic weighted mark method was used to analyze the comprehensive effect of UPC content on coke quality. The results show that because of the decrease of the content of intermediates Cf(O) and C(O)Cf(O) the restrictive step between the coke and CO2 is an interfacial chemical reaction, and it accords with the Mckewan equation 1-(1-α)1/3=kt. The UPC has a strong effect on coke when the content of UPC is 10~20%; meanwhile , the CRI and the apparent porosity are significantly decreased by 6.8% and 9.5%, respectively, and the CSR is significantly increased by 3.8%. The UPC can effectively reduce the effect of CO2 on the edge and the internal erosion of coke; the large pores and the pulverization of coke were avoided. The results of the synthetic weighted mark method showed that the comprehensive quality of coke changed greatly when the content of unburned pulverized coal was 11.24~20.87%, which is in agreement with the experimental results.

Ming-Ming Li,* Jia Cao,* Jia-Chun Yang, Yu-Jie Shen, Xiao-Lei Cai, Yuan-Wen Chen, Chun-Ying Qu, Yi Zhang, Feng Shen, Lei-Ming Xu Department of Gastroenterology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: Quantum dots (QDs) conjugated with integrin antagonist arginine–glycine–aspartic acid (RGD) peptides (QDs-RGD) are novel nanomaterials with a unique optical property: a high molar extinction coefficient. Previously, we have shown that QDs-RGD demonstrate a photodynamic therapy (PDT) effect as new photosensitizers for the pancreatic cancer cell line SW1990 in vitro. Here, we investigate the application of QDs-RGD in mice bearing pancreatic tumors using PDT. To ensure that more photosensitizers accumulated in tumors, QDs-RGD were injected intratumorally. After selection of an adequate dosage for injection from analyses of biodistribution images captured by an IVIS system, PDT was initiated. Three groups were created according to different PDT procedures. In group 1, mice were injected with QDs-RGD intratumorally, and an optical fiber connected to a laser light was inserted directly into the tumor. Irradiation was sustained for 20 min with a laser light (630 nm) at 100 mW/cm2. In group 2, the laser optical fiber was placed around, and not inserted into, tumors. In group 3, PDT was conducted as in group 1 but without injection of QDs-RGD. After 28 days of observation, tumors on the back of mice in group 1 grew slowly (V/V0 =3.24±0.70) compared with the control groups, whose tumors grew quickly, and the mean V/V0 reached 6.08±0.50 (group 2) and 7.25±0.82 (group 3). Histology of tumor tissues showed more necrotic tissues, more inflammatory cells, and less vascular tissue in the PDT group than those in the control groups. These results suggest that QDs-RGD-mediated PDT, with illumination using an optical fiber inserted directly into the tumor, can inhibit the growth of SW1990 tumors with high efficiency in nude mice. Keywords: quantum dots, RGD peptides, pancreatic neoplasm, intratumoral injection, photodynamic therapy

Chun-Ying Qu,1,* Min Zhou,1,* Ying-wei Chen,2 Mei-mei Chen,3 Feng Shen,1 Lei-Ming Xu11Digestive Endoscopic Diagnosis and Treatment Center, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China; 2Shanghai Key Laboratory of Pediatric Gastroenterology and Nutrition, Shanghai, People’s Republic of China; 3Digestive Department, Xinhua Hospital, School of medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China*These authors contributed equally to this workPurpose: The first-line chemotherapy treatment protocol for gastric cancer is combination chemotherapy of 5-fluorouracil (5-FU) and cisplatin (CDDP). The aim of this study was to engineer prodrug-based nanostructured lipid carriers (NLC) platform for codelivery of 5-FU and CDDP to enhance therapy and decrease toxicity.Methods: First, 5-FU-stearic acid lipid conjugate was synthesized by two steps. Second, 5-FU-stearic acid prodrug and CDDP were loaded in NLC. Finally, hyaluronic acid (HA) was coated onto NLC surface. Average size, zeta potential, and drug loading capacity of NLC were evaluated. Human gastric cancer cell line BGC823 (BGC823 cells) was used for the testing of in vitro cytotoxicity assays. In vivo antitumor activity of NLC was evaluated in mice bearing BGC823 cells model.Results: HA-coated 5-FU-stearic acid prodrug and CDDP-loaded NLC (HA-FU/C-NLC) showed a synergistic effect in combination therapy and displayed the greatest antitumor activity than all of the free drugs or uncoated NLC in vitro and in vivo.Conclusion: This work reveals that HA-coated NLC could be used as a novel carrier to codeliver 5-FU and CDDP for gastric cancer therapy. HA-FU/C-NLC could be a promising targeted and combinational therapy in nanomedicine.Keywords: gastric cancer, nanostructured lipid carriers, hyaluronic acid, combination chemotherapy, lipid prodrug

Fei Shen,1 Wen-Song Cai,1 Jiang-Lin Li,1 Zhe Feng,1 Qi-cai Liu,2 Huan-qing Xiao,1 Jie Cao,1 Bo Xu1 1Department of General Surgery, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou, People's Republic of China; 2Experimental Medical Research Center, Guangzhou Medical University, Guangzhou, People's Republic of China Abstract: Liver metastasis is a major cause of mortality in colorectal cancer (CRC). The current study was to investigate the ability of ulinastatin (UTI) and curcumin (CUR) to inhibit CRC liver metastases via modulating matrix metalloproteinase-9 (MMP-9) and E-cadherin expression. Human CRC HCT-116 cells were treated with compounds individually and in combination in order to understand the effect on cell migration and invasion. The HCT-116 cell line was established to stably express luciferase and green fluorescent protein (GFP) by lentiviral transduction (HCT-116-Luc-GFP). We identified an anti-metastasis effect of UTI and CUR on a CRC liver metastasis mouse model. Tumor development and therapeutic responses were dynamically tracked by bioluminescence imaging. Expression of MMP-9 and E-cadherin in metastatic tumors was detected by immunohistochemical assay. Results of wound healing and cell invasion assays suggest that treatment with UTI, CUR, and UTI plus CUR, respectively, significantly inhibit HCT-116 cell migration and invasion. Furthermore, results of CRC hepatic metastasis on a nude mouse model showed that treatment with UTI, CUR alone, and a combination notably inhibited hepatic metastases from CRC and prolonged survival of tumor-bearing mice, especially in the UTI plus CUR group. These results suggest that the combination of UTI and CUR together may offer greater inhibition against metastasis of CRC. Keywords: hepatic metastasis, bioluminescence imaging, therapy

Fei Shen, Jiang-Lin Li, Weng-Song Cai, Guang-Hui Zhu, Wei-Li Gu, Lin Jia, Bo XuDepartment of General Surgery, Guangzhou First People's Hospital, Guangzhou Medical College, Guangzhou, People's Republic of ChinaAbstract: Interleukin (IL)-12 has emerged to be a prospective molecule for antitumor therapies with many types of cancers. Here we examine the effect of IL-12 treatment in preventing the colorectal cancer liver metastasis in a mice model. At different doses, we found that IL-12 treatment decreased the formation of liver metastasis sites and the percentage of metastasis volume in the liver. Additionally, this treatment leads to improved survival function of mice after tumor cell transplantation. We believe that IL-12 based therapy provided a novel treatment to colorectal cancer patients suffering from liver metastasis.Keywords: IL-12, liver metastasis, colorectal cancer, therapy

Solid-state lithium batteries are promising for safety and energy density compared with traditional lithium-ion batteries. However, the large interfacial resistance between the electrode and electrolyte is a bottleneck to achieving high-performance solid-state batteries. Engineered electrode structures with a porous scaffold of the solid electrolyte material are promising to lower the interfacial resistance and provide a mechanical support for a thin solid electrolyte layer. In this work, two ceramic processing techniques are used to fabricate porous/dense bilayer architectures based on a Li6.25Al0.25La3Zr2O12 (LLZO) Li-garnet material. Finger-like vertically aligned pores are created by the phase inversion (PI) process. A water bath presaturated with Li salt prevents Li loss during the PI solvent exchange step. Pore size and porosity can be optimized by adjusting the bath temperature. The high shear compaction process was used to prepare LLZO tapes with 40, 60, and 80 vol% poreformer. The porosity of the tapes after sintering is 39.5%, 58.4%, and 75.4%, respectively. Microtomography exhibits the porosity, pore shape, and pore distribution of the tapes. A typical cathode material LiNi0.33Mn0.33Co0.33O2 (NMC) is filled into the pores via vacuum infiltration, and a dense cathode layer is formed within the garnet scaffold.