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Candida species are the most commonly isolated opportunistic fungal pathogens in humans. Candida albicans causes most of the diagnosed infections, closely followed by Candida glabrata. C. albicans is well studied, and many genes have been shown to be important for infection and colonization of the host. It is however less clear how C. glabrata infects the host. With the help of fungal RNA enrichment, we here investigated for the first time the transcriptomic profile of C. glabrata during urinary tract infection (UTI) in mice. In the UTI model, bladders and kidneys are major target organs and therefore fungal transcriptomes were addressed in these organs. Our results showed that, next to adhesins and proteases, nitrogen metabolism and regulation play a vital role during C. glabrata UTI. Genes involved in nitrogen metabolism were upregulated and among them we show that DUR1,2 (urea amidolyase) and GAP1 (amino acid permease) were important for virulence. Furthermore, we confirmed the importance of the glyoxylate cycle in the host and identified MLS1 (malate synthase) as an important gene necessary for C. glabrata virulence. In conclusion, our study shows with the support of in vivo transcriptomics how C. glabrata adapts to host conditions.

Xiaofang Wen,1,&ast; Nan Zeng,1,&ast; Ningbo Zhang,1,2,&ast; Tingting Ou,1 Xiaowei Li,1 Xiaoying Li,1 Wangen Li,1 Kang Xu,3 Tao Du1 1Department of Endocrinology, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510260, People’s Republic of China; 2Department of Endocrinology, Peking University Shenzhen Hospital, Shenzhen, 518036, People’s Republic of China; 3Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Tao Du; Kang Xu, Email dutao@gzhmu.edu.cn; xukang@mail.sysu.edu.cnBackground: Although flash continuous glucose monitoring systems (FCGM) accuracy has been extensively studied in diabetes, its accuracy is still not fully evaluated in type 2 diabetes (T2D) patients in real-world settings. In the present study, we aim to assess the effects of diabetes complications and related comorbidities on FCGM accuracy in T2D patients with diabetes complications and related comorbidities in the real world.Methods: FCGM data were collected at eight-time points daily (3 AM, 7 AM, 9 AM, 11 AM, 1 PM, 5 PM, 7 PM, and 9 PM) from 742 patients with T2D and compared with simultaneous fingertip capillary blood glucose (reference blood glucose, REF), and the difference was evaluated using Parkes error grid (PEG), surveillance error grid (SEG), and logistic regression analysis.Results: In total, 25,579 FCGM/REF data pairs were included in the study. The FCGM values were lower than the paired REF values in 75% of the pairs. The maximum bias (− 23.0%) and maximum mean absolute relative difference (24.5%) were observed at 3 AM among eight-time points. SEG analysis also demonstrated the highest percentage of paired readings in moderate and great risk zone (C and D) at 3 AM than PEG analysis (7.33% vs 0.43%, P< 0.001). According to the SEG classification, hypoglycemia, infection, diabetic foot, diabetic ketoacidosis, and hypertension were independent risk factors that impaired FCGM accuracy in patients.Conclusion: FCGM commonly underestimates blood glucose levels. Compared with PEG, SEG analysis seems more conducive to the analysis of FCGM performance. The present data highlights the impairment of diabetes complications and related comorbidities on the FCGM accuracy in T2D patients.Keywords: flash continuous glucose monitoring system, type 2 diabetes, surveillance error grid, Parkes error grid, diabetes complications

Jing Wang,1 Shuting Zuo,1 Yan Zhang,1 Shanzhi Li,2 Ying Shi,1 Tonghua Du,1 Jicheng Han,2 Ningyi Jin,2– 4 Yiquan Li,2 Xiao Li2,3 1Department of Breast Surgery, The Second Hospital of Jilin University, Changchun, 130000, People’s Republic of China; 2Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun, 130117, People’s Republic of China; 3Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, 130122, People’s Republic of China; 4Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, People’s Republic of ChinaCorrespondence: Yiquan Li; Xiao Li, Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Liuying West Road, 666, Jingyue Economic & Technological Development Zone, Changchun, People’s Republic of China, Tel +86-431-86985923, Fax +86-431-87985861, Email 905381255@qq.com; 1077614276@qq.comPurpose: Oncolytic virus therapy has gradually become an integral approach in cancer treatment. We explored the therapeutic effects of the combination of a dual cancer-selective anti-tumor recombinant adenovirus (Ad-Apoptin-hTERTp-E1a) and cyclophosphamide on breast cancer cells.Methods: The inhibition of MCF-7 and MDA-MB-231 breast cancer cells by Ad-Apoptin-hTERTp-E1a (Ad-VT), cyclophosphamide, and Ad-VT + Cyclophosphamide was investigated using the CCK-8 assay. The combination index (CI) was calculated using CalcuSyn software to determine the best combination based on the inhibition rates of the different treatment combinations. The CCK-8 assay and crystal violet staining were used to detect the cytotoxicity of the combined Ad-VT and cyclophosphamide in breast cancer cells and breast epithelial cells. Subsequently, Hoechst staining, annexin V flow cytometry, and JC-1 staining were used to analyze the inhibitory pathway of Ad-VT plus cyclophosphamide on breast cancer cells. Cell migration and invasion of breast cancer cells were assessed using the cell-scratch and Transwell assays. The anti-tumor effects of different treatment groups in a tumor-bearing nude mouse model also were analyzed.Results: The treatment combination of Ad-VT (40 MOI) and cyclophosphamide (400 μM) significantly inhibited MCF-7 and MDA-MB-231 cells and reduced the toxicity of cyclophosphamide in normal cells. Ad-VT primarily induced breast cancer cell apoptosis through the endogenous apoptotic pathway. Apoptosis was significantly increased after treatment with Ad-VT plus cyclophosphamide. The combination significantly inhibited the migration and invasion of MCF-7 and MDA-MB-231 cells. The in vivo experiments demonstrated that exposure to Ad-VT plus cyclophosphamide significantly inhibited tumor growth and extended the survival time of the nude mice.Conclusion: Ad-VT plus cyclophosphamide reduced toxicity and exhibited increased efficacy in treating breast cancer cells.Keywords: cyclophosphamide, recombinant adenovirus, breast cancer, synergy, toxicity

Li Li,1 Rongjin Sun,1 Joseph Zenga,2 Heather Himburg,3 Lu Wang,1 Shengnan Duan,1 Jingwen Liu,1 Dinh Bui,1 Zuoxu Xie,1 Ting Du,4 Lijun Xie,1 Taijun Yin,1 Stu Wong,2 Song Gao,4 Ming Hu1 1Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA; 2Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, Milwaukee, WI, USA; 3Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, WI, USA; 4Department of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX, USACorrespondence: Ming Hu, Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA, Email mhu@uh.edu Song Gao, Department of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX, USA, Email Song.Gao@tsu.eduObjective: We aim to quantify the absolute protein expression of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in various cells and tissues to determine the relative contribution of COX-1 and COX-2 to PGE2 production.Methods: An LC-MS method was developed and validated, then used for quantifying the absolute amounts of COX-1 and COX-2 in recombinant human COX-1 and COX-2, lysates from different cells, tissue microsomes of rodents and humans, Pirc rat colonic polyps, and biopsy specimens from squamous cell carcinoma (SCC) patients. The COX-1 and COX-2 turnover numbers were subsequently calculated based on apparent formation rates of PGE2.Results: A robust LC-MS method for quantification of COX-1 and COX-2 was developed and validated and then used to calculate their apparent turnover numbers. The results showed that COX-1 expression levels were much higher than that of COX-2 in all the tested tissues including the colonic epithelium of F344 (28-fold) and Pirc rats (20-fold), colonic polyps of Pirc rats (8-fold), and biopsy specimens of SCC patients (11– 17-fold). In addition, both COX-1 and COX-2 were higher in polyps when compared to adjacent mucosa of Pirc rats. The turnover number of recombinant human COX-2 was 14-fold higher than that of recombinant human COX-1. LPS stimulation increased COX-2 protein expression in three cell lines (Raw 264.7, SCC9 and EOMA) as expected but unexpectedly increased COX-1 protein expression (13.8-fold) in EOMA cells.Conclusion: In human oral cancer tissues and cells as well as Pirc rat colon, COX-1 plays an unexpectedly but more important role than COX-2 in abnormal PGE2 production since COX-1 expression is much higher than COX-2. In addition, COX-1 expression levels are inducible in cells, and higher in polyps than surrounding mucosa in Pirc rat colon. These results indicate that targeted suppression of local COX-1 should be considered to reduce colon-specific PGE2-mediated inflammation.Graphical Abstract: Keywords: cyclooxygenase-1, cyclooxygenase-2, turnover number, inflammation, absolute quantification, liquid chromatography with tandem mass spectrometry

Abstract Extracellular vesicles (EV) are membranous particles secreted by all cells and found in body fluids. Established EV contents include a variety of RNA species, proteins, lipids and metabolites that are considered to reflect the physiological status of their parental cells. However, to date, little is known about cell‐type enriched EV cargo in complex EV mixtures, especially in urine. To test whether EV secretion from distinct human kidney cells in culture differ and can recapitulate findings in normal urine, we comprehensively analysed EV components, (particularly miRNAs, long RNAs and protein) from conditionally immortalised human kidney cell lines (podocyte, glomerular endothelial, mesangial and proximal tubular cells) and compared to EV secreted in human urine. EV from cell culture media derived from immortalised kidney cells were isolated by hydrostatic filtration dialysis (HFD) and characterised by electron microscopy (EM), nanoparticle tracking analysis (NTA) and Western blotting (WB). RNA was isolated from EV and subjected to miRNA and RNA sequencing and proteins were profiled by tandem mass tag proteomics. Representative sets of EV miRNAs, RNAs and proteins were detected in each cell type and compared to human urinary EV isolates (uEV), EV cargo database, kidney biopsy bulk RNA sequencing and proteomics, and single‐cell transcriptomics. This revealed that a high proportion of the in vitro EV signatures were also found in in vivo datasets. Thus, highlighting the robustness of our in vitro model and showing that this approach enables the dissection of cell type specific EV cargo in biofluids and the potential identification of cell‐type specific EV biomarkers of kidney disease.

The rapidly increasing production and application of graphene oxide (GO) calls for much-needed attention to aspects related to the environmental implications of this unique nanomaterial. To date, very little is known about the transformation of GO when exposed to electromagnetic radiation at different wavelengths, which can occur in natural water systems. This study explored the changes in the physicochemical properties of GO when exposed to the UV and visible wavelength ranges of simulated solar light. Irradiation, especially under UV light, led to remarkable changes in the surface oxygen (O)-functionalities of GO, and cleavage of GO's graphitic bond. The carbon/oxygen (C/O) ratio of GO increased by 41% and 31% after irradiation with simulated solar light and UV irradiation, respectively. Visible light also changed the morphology of GO, but the C/O ratio only increased by 5.5%. The formation of hydroxyl radicals ( OH), superoxide (O2−) and singlet oxygen (1O2) by GO was observed during irradiation with both UV and visible lights, but the proportions were different. 1O2 was primarily formed during visible light irradiation, while O2− and OH were mostly formed under UV light. OH hydroxylated GO, which mainly resulted in the decomposition of GO. The changes in O-containing functional groups on GO by irradiation influenced the colloidal stability of GO in aqueous media, but the physical wrinkle on GO surface may strongly affect its interactions with other organic materials.

The development of optical quality GeO2–TiO2–SiO2 glasses compatible with direct ink writing (DIW) 3D printing is reported in this study. Colloidal GeO2 and TiO2–SiO2 core–shell feedstocks are prepared by a sol–gel method and converted to printable inks for DIW. Printed inks are subsequently densified to glass using heat treatment at temperatures up to 1100 °C in air. A series of print‐compatible glass compositions are prepared, yielding transparent glass with the highest refractive index n = 1.576. Herein, it is shown that more TiO2 can be incorporated into the glass without haziness or scattering through the addition of GeO2. The mechanisms for this are discussed, and it is shown that crystallization persists in these glasses despite the lack of visible light scattering. Finally, the combination of both TiO2 and GeO2 into a ternary glass also is used to independently tune the dispersion properties of the glass, as demonstrated by measured refractive indices and Abbe numbers, which has potential implications for 3D‐printed optics.

Mingbiao Ma,1– 3,&ast; Minjun Chu,1,2,&ast; Lvyan Tao,3– 5,&ast; Jue Li,1,2 Xiaojuan Li,1,2 Hailin Huang,1,2 Kexuan Qu,6 Haiping Wang,1,2 Li Li,3– 5 Tingyi Du1– 3 1Department of Clinical Laboratory, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of China; 2Department of Clinical Laboratory, Children’s Hospital Affiliated to Kunming Medical University, Kunming, Yunnan, People’s Republic of China; 3Yunnan Key Laboratory of Children’s Major Disease Research, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of China; 4Yunnan Institute of Pediatrics, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of China; 5Kunming Key Laboratory of Children Infection and Immunity, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of China; 6Department of Blood Transfusion, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Li LiYunnan Institute of Pediatrics, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of ChinaEmail lili@etyy.cnTingyi DuDepartment of Clinical Laboratory, Kunming Children’s Hospital, Kunming, Yunnan, People’s Republic of ChinaEmail dutingyi@etyy.cnPurpose: The present study investigated the prevalence characteristics of oxacillin susceptible mecA-positive Staphylococcus aureus (OS-MRSA) in a children’s hospital in Kunming from January 2019 to December 2020.Methods: A total of 499 S. aureus strains were included in the study and tested for oxacillin susceptibility using the VITEK 2 Compact automated antimicrobial susceptibility test system. All oxacillin-susceptible strains were detected mecA and mecC by polymerase chain reaction (PCR). E-test was used to compare the minimum inhibitory concentration (MIC) values of methicillin-susceptible S. aureus (MSSA), methicillin-resistant S. aureus (MRSA), and OS-MRSA for oxacillin, cefoxitin, penicillin, vancomycin, erythromycin, and clindamycin. Molecular typing of OS-MRSA was performed by MLST and SCCmec typing. Toxin genes were detected by PCR.Results: Forty-five OS-MRSA strains were detected, for an overall rate of 9.02% (45/499). The MICs of MSSA, OS-MRSA, and MRSA against oxacillin were concentrated at 0.38, 0.38, and 12 μg/mL, respectively; the cefoxitin MICs of MSSA and MRSA were concentrated at 2 and 32 μg/mL respectively; and MICs of OS-MRSA were concentrated at 2 and 8 μg/mL; penicillin, vancomycin and erythromycin MICs against MSSA, OS-MRSA, and MRSA showed same centralized points and were 32, 1, and 256 μg/mL, respectively; the MICs of clindamycin against MSSA were 0.5 μg/mL, while that against OS-MRSA and MRSA were concentrated at 256 μg/mL. Molecular typing of OS-MRSA was dominated by ST59-SCCmec IV. The carrier rates of hemolysin genes (hl-a, hl-d) and fibrinogen-binding clumping factor genes (clfA, clfB) were 100% in OS-MRSA, followed by 40% (18/45) for enterotoxin genes (sea, seb).Conclusion: OS-MRSA has a high detection rate in children, and main molecular typing is ST59-SCCmecIV in Kunming. The identification ability of automated antibacterial drug sensitivity test detection systems for OS-MRSA is very limited. A combination of phenotypic analysis and molecular detection should be used to improve OS-MRSA identification.Keywords: Staphylococcus aureus, MRSA, OS-MRSA, children, oxacillin

Shenni Yi,1,* Zhan Li,1,* Xiuqin Wang,1,* Tiantian Du,1 Xiuhong Chu2 1Department of Respiratory and Critical Medicine, Yantai Yuhuangding Hospital, Yantai, Shandong Province, 264000, People’s Republic of China; 2Department of Nursing, Yantai YEDA Hospital, Yantai, Shandong Province, 264006, People’s Republic of China*These authors contributed equally to this workCorrespondence: Tiantian DuDepartment of Respiratory and Critical Medicine, Yantai Yuhuangding Hospital, East Yuhuangding Road NO. 20, Yantai, Shandong Province, 264000, People’s Republic of ChinaEmail quebi7192@163.comXiuhong ChuDepartment of Nursing, Yantai YEDA Hospital, Taishan Road No. 11, Yantai Development Area, Yantai, Shandong Province, 264006, People’s Republic of ChinaEmail vkr758157@163.comBackground: Circular RNAs (circRNAs) are non-coding RNAs with a covalently closed loop. circRNAs affect the progression of diverse cancers. Nonetheless, circ_0001806 expression and function in non-small cell lung cancer (NSCLC) are undefined.Methods: qRT-PCR was executed to examine circ_0001806, miR-1182 and NOVA2 mRNA expression levels in NSCLC tissues and cells. CCK-8, EdU, cell scratch test and Transwell assay were conducted to examine the viability, multiplication, migration and invasion of NSCLC cell lines H1650 and HCC827. The binding sites between circ_0001806 and miR-1182, miR-1182 and NOVA2 mRNA were predicted by the circular RNA Interactome and TargetScan databases, and the dual-luciferase reporter gene experiment was employed for verification. Western blot was implemented to examine NOVA2 expression.Results: Circ_0001806 expression in NSCLC tissues and cell lines was substantially augmented, while miR-1182 expression was markedly decreased. Circ_0001806 facilitated the multiplication, migration and invasion of H1650 and HCC827 cells, while miR-1182 exerted the opposite effect. Circ_0001806 indirectly enhanced NOVA2 expression by specifically down-modulating miR-1182.Conclusion: Circ_0001806 augments NOVA2 expression by targeting miR-1182 to enhance the multiplication, migration and invasion of NSCLC cells.Keywords: circ_0001806, miR-1182, NOVA2, NSCLC

Lele Li,1,* Tiantian Du,2,* Chi Zhang3 1School of Public Policy and Management, Tsinghua University, Beijing 100084, People’s Republic of China; 2Institute for Hospital Management, Tsinghua University, Shenzhen 518055, People’s Republic of China; 3Chinese Academy of Fiscal Sciences, Beijing 100142, People’s Republic of China*These authors contributed equally to this workCorrespondence: Lele LiSchool of Public Policy and Management, Tsinghua University, No.1 Tsinghua Yard, Haidian District, Beijing 10084, People’s Republic of ChinaTel +86-010-6147-2626Email lilele1211@163.comBackground: Air pollution is an important factor in health outcomes and health-care expenditure. It has become an important issue of global concern. The objective of this study was to explore the influence of air pollution on the economic burden of respiratory diseases using different levels of PM2.5.Methods: Starting from the demand side, we took the 3,546 patients in the Respiratory and Critical Care Department of a tertiary hospital in Beijing between 2013 and 2015 as examples, combining daily air-quality data using a generalized linear regression–analysis model to explore the impact of air pollution on health-care expenditure on a microindividual level.Results: We found that PM2.5 had a significant impact on health-care expenditure on respiratory diseases. It had a positive impact on total health-care expenditure, drug expenditure, and antibiotic expenditure. The impact of different levels of air pollution on the health care–expenditure burden of disease was heterogeneous. As the air-pollution index increased, health care–expenditure burden of respiratory diseases also gradually raised. The impact of PM10 and air-quality index had a positive impact on health-care expenditure for respiratory diseases. Air pollution had a significant impact on health care–expenditure burden of respiratory diseases. The effect of length of stay on various health-care expenditure was significantly positive.Conclusion: The impact of mortality-risk classification on various health-care expenditure is significant. Therefore, policy-making must take into account both the supply side and the demand side of health-care services. Furthermore, the government should strengthen environmental governance, pay attention to the heterogeneity of the health care–expenditure burden affected by environmental pollution, improve the medical insurance system, and improve the health of residents to reduce the health care–expenditure burden.Keywords: air pollution, PM2.5, respiratory diseases, health care–expenditure burden, individual health level

Lele Li,1 Tiantian Du,2 Yanping Hu3 1School of Public Policy and Management, Tsinghua University, Beijing 100084, People’s Republic of China; 2Institute for Hospital Management, Tsinghua University, Shenzhen City 518055, Guangdong Province, People’s Republic of China; 3Department of Medical Engineering, China-Japan Friendship Hospital, Beijing 100029, People’s Republic of ChinaCorrespondence: Yanping Hu Email nd3388@sina.comBackground: With population aging (PA), the healthcare expenditure (HE) increases. The aim of this study is to analyze the HE of different age groups and the effect of age on HE among different age groups.Methods: Combining PA and HE data, this study used the fixed effect model and parameter estimation method to evaluate the influence of different age groups on HE from 2008 to 2014.Results: The age effect of HE for the population aged 65 or over was the most significant among the different age groups. Based on PA and HE data, HE per capita of the age group 65 years or over is 7.25 times as much as the population aged < 25 years, 1.61 times as much as the population aged 25∼ 59 years, and 3.47 times as much as the population aged 60∼ 64 years. Based on the result of the fixed effect model, HE per capita of the age group < 25 years was 218.39 Yuan (CNY) (USD $31.2). HE per capita of the age group 25∼ 59 years old increased to 1,548.62 Yuan (CNY) (USD $221.2). HE per capita of the 60∼ 64 years age group will be 921.56 Yuan (CNY) (USD $131.7), 4.22 times as much as that of the age group < 25 years. HE per capita in the age group of 65 years or over is 2,538.88 Yuan (CNY) (USD $362.7), 11.63 times as much as that of the age group < 25 years.Conclusion: The results suggest that PA in China is intensifying. In order to control the rising of HE, the government should not only address the supply side such as reforming medical insurance payment, developing new technologies, but also focusing on solving the demand side such as improving the quality of healthcare services, solving environmental pollution, and improving the residents’ health.Keywords: population aging, healthcare demand, healthcare expenditure, different age groups, fixed effect model, parameter estimation method

Shufen Xu,* Shengping Zhai,* Tiantian Du,* Zhan Li Respiratory Department, Yantai Yuhuangding Hospital Affiliated to Qingdao University, Yantai City, Shandong Province 264000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhan LiRespiratory Department, Yantai Yuhuangding Hospital Affiliated to Qingdao University, NO. 20 Yuhuangding East Road, Zhifu District, Yantai City, Shandong Province 264000, People’s Republic of ChinaEmail gvjycert0951@126.comIntroduction: LncRNA MIR503HG has been reported to participate in liver cancer and ALK-negative anaplastic large-cell lymphoma, while its role in non-small cell lung cancer (NSCLC) is unknown. We therefore investigated the functions of lncRNA MIR503HG in NSCLC.Methods: MIR503HG expression in paired cancer and non-cancer tissues from NSCLC patients was analyzed by RT-qPCR. The interaction between cyclin D1 and MIR503HG was analyzed by overexpression experiments. Cell cycle analysis was performed by flow cytometry. Cell proliferation was analyzed by CCK-8 assay.Results: MIR503HG was downregulated in NSCLC and low levels of MIR503HG were associated with poor survival. In contrast, cyclin D1 was upregulated in NSCLC, and cyclin D1 and MIR503HG were inversely correlated. In NSCLC cells, overexpression experiments revealed that MIR503HG functioned as an upstream inhibitor of cyclin D1. MIR503HG overexpression led to G1 cell cycle arrest, while overexpression of cyclin D1 attenuated the effects of MIR503HG overexpression. Similarly, MIR503HG overexpression resulted in reduced cell proliferation rate, while overexpression of cyclin D1 caused the increased cell proliferation rate and attenuated effects of MIR503HG overexpression.Conclusion: MIR503HG inhibits NSCLC cell proliferation by inducing cell cycle arrest through the downregulation of cyclin D1.Keywords: non-small cell lung cancer, lncRNA MIR503HG, cyclin D1, cell cycle, survival

As part of the human microbiota, the fungus Candida albicans colonizes the oral cavity and other mucosal surfaces of the human body. Commensalism is tightly controlled by complex interactions of the fungus and the host to preclude fungal elimination but also fungal overgrowth and invasion, which can result in disease. As such, defects in antifungal T cell immunity render individuals susceptible to oral thrush due to interrupted immunosurveillance of the oral mucosa. The factors that promote commensalism and ensure persistence of C. albicans in a fully immunocompetent host remain less clear. Using an experimental model of C. albicans oral colonization in mice we explored fungal determinants of commensalism in the oral cavity. Transcript profiling of the oral isolate 101 in the murine tongue tissue revealed a characteristic metabolic profile tailored to the nutrient poor conditions in the stratum corneum of the epithelium where the fungus resides. Metabolic adaptation of isolate 101 was also reflected in enhanced nutrient acquisition when grown on oral mucosa substrates. Persistent colonization of the oral mucosa by C. albicans also correlated inversely with the capacity of the fungus to induce epithelial cell damage and to elicit an inflammatory response. Here we show that these immune evasive properties of isolate 101 are explained by a strong attenuation of a number of virulence genes, including those linked to filamentation. De-repression of the hyphal program by deletion or conditional repression of NRG1 abolished the commensal behaviour of isolate 101, thereby establishing a central role of this factor in the commensal lifestyle of C. albicans in the oral niche of the host.

Tan Du, 1,* Xingyuan Jia, 2,* Xichen Dong, 2 Xiaoli Ru, 3 Lina Li, 2 Yakun Wang, 2 Jian Liu, 2 Guosheng Feng, 1 Tao Wen 2 1Department of Oncology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, People’s Republic of China; 2Medical Research Center, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, People’s Republic of China; 3Department of Gynecology and Obstetrics Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, People’s Republic of China*These authors contributed equally to this workCorrespondence: Guosheng FengDepartment of Oncology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, People’s Republic of ChinaTel +86 10 85231569Fax +86 10 85231407Email fgscy010@163.comTao WenMedical Research Center, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, People’s Republic of ChinaTel +86 10 85231154Email wentao5281@163.comBackground: Breast cancer remains the most lethal malignancy in women worldwide. Aberrant O-glycosylation is closely related to many human diseases, including breast carcinoma; however, its precise role in cancer development is insufficiently understood. Cosmc is an endoplasmic reticulum-localized chaperone that regulates the O-glycosylation of proteins. Cosmc dysfunction results in inactive T-synthase and expression of truncated O-glycans such as Tn antigen. Here we investigated the impact of Cosmc disruption-mediated aberrant O-glycosylation on breast cancer cell development through in vitro and in vivo experiments.Materials and Methods: We deleted the Cosmc gene in two breast cancer cell lines (MCF7, T47D) using the CRISPR/Cas-9 system and then measured the expression levels of Tn antigen. The proliferation of Tn-positive cells was examined by RTCA, colony formation and in vivo experiments. The effects of Cosmc deficiency on glycoprotein CD44 and MAPK pathway were also determined.Results: Both in vitro and in vivo studies showed that Cosmc deficiency markedly suppressed breast cancer cell growth compared with the corresponding controls. Mechanistically, Cosmc disruption impaired the protein expression of CD44 and the associated MAPK signaling pathway; the latter plays a crucial role in cell proliferation. Reconstitution of CD44 substantially reversed the observed alterations, confirming that CD44 requires normal O-glycosylation for its proper expression and activation of the related signaling pathway.Conclusion: This study showed that Cosmc deficiency-mediated aberrant O-glycosylation suppressed breast cancer cell growth, which was likely mediated by the impairment of CD44 expression.Keywords: breast cancer, O-glycosylation, Tn antigen, tumor growth

Taofeng Du,1,2,* Guang Zhu,1,2,* Xiaoping Wu,1,2 Junyang Fang,1,2 En-Min Zhou1,2 1Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, People’s Republic of China; 2Scientific Observing and Experimental Station of Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, Yangling 712100, Shaanxi, People’s Republic of China*These authors contributed equally to this workCorrespondence: En-Min ZhouDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, People’s Republic of ChinaEmail zhouem@nwsuaf.edu.cnBackground: Enzyme-linked immunosorbent assay (ELISA) is a common method for diagnosing swine influenza. However, the production of classical antibodies is both costly and time-consuming. As a promising alternative diagnostic tool, single-domain antibodies (sdAbs) offer the advantages of simpler and faster generation, good stability and solubility, and high affinity and specificity.Methods: Phage display technology was used to isolate sdAbs against the SIV-NP protein from a camel VHH library. The sdAb5 was fused to the biotin acceptor peptide (BAP) and a His-Tag for its expression as monomeric and site-specific biotinylation in E.coli to develop an sdAb-based blocking ELISA (sdAb-ELISA). In the sdAb-ELISA, the anti-SIV antibodies from swine samples were used to block the binding between the biotinylated sdAb5 and SIV-NP protein coated on the ELISA plate. The specificity, sensitivity, and reproducibility of sdAb-ELISA were determined. In addition, consistency among sdAb-ELISA, commercial ELISA kit, and Western blot was evaluated.Results: Six SIV-NP-specific sdAbs were isolated, among which sdAb5 was identified as a dominant sdAb with higher reactivity. The cut-off value of biotinylated sdAb5-based bELISA was determined to be 29.8%. Compared with the positive reference serum against five different types of swine viruses, the developed sdAb-ELISA showed 100% specificity. The detection limit of sdAb-ELISA was 1:160 in an anti-SIV positive reference serum, which is lower than that of the commercial ELISA kit (1:20). In 78 diluted anti-SIV positive serum (1:80), 21 and 42 samples were confirmed as positive by the commercial ELISA kit and sdAb-ELISA, respectively. The coefficients of variation of intra- and inter-assay were 1.79–4.57% and 5.54–9.98%, respectively. The sdAb-ELISA and commercial ELISA kit showed a consistency of 94.17% in clinical swine serum samples. Furthermore, the coincidence rate was 96.67% between the results detected by sdAb-ELISA and Western blot.Conclusion: A specific, sensitive, and reproducible sdAb-ELISA was successfully developed, which offers a new, promising method to detect anti-SIV antibodies in swine serum.Keywords: SIV, NP, sdAb, sdAb-ELISA

Qianqian Zhang,1,2,* Meimei Xiong,2,* Jinlu Liu,1,2 Shuai Wang,2 Ting Du,2 Tianyi Kang,2 Yu Liu,2 Hao Cheng,2 Meijuan Huang,1 Maling Gou21Department of Thoracic Oncology, West China Hospital, Sichuan University, Chengdu, 610041, People’s Republic of China; 2State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, People’s Republic of China*These authors contributed equally to this workBackground: Phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPP) is a novel tumor suppressor. However, whether LHPP is effective to melanoma has not been investigated. Gene therapy provides a new strategy for the treatment of melanoma. Currently, it suffers from the lack of safe and effective gene delivery systems. Methods: A CRGDKGPDC peptide (iRGD) modified hybrid monomethoxy poly(ethylene glycol)-poly(D,L-lactide) nanoparticle (iDPP) was prepared and complexed with a LHPP plasmid, forming an iDPP/LHPP nanocomplex. The iDPP/LHPP nanocomplex was characterized by particle size distribution, zeta potential, morphology, cytotoxicity, and transfection efficiency. The antitumor efficacy of the nanocomplex against melanoma was studied both in vitro and in vivo. Further, the potential epigenetic changes in melanoma induced by iDPP/LHPP nanocomplex were evaluated.Results: The iDPP/LHPP nanocomplex showed high transfection efficiency and low toxicity. Moreover, the nanocomplex displayed a neutral charge that can meet the requirement of intravenous injection for targeted gene therapy. In vitro and in vivo experiments indicated that the iDPP/LHPP nanocomplex significantly inhibited the melanoma growth without causing notable adverse effects. We also found that LHPP played an important role in epigenetics. It regulated the expression of genes related to the proliferation and apoptosis chiefly at the level of transcription.Conclusion: This work demonstrates that the iDPP nanoparticle-delivered LHPP gene has a potential application in melanoma therapy through regulation of the genes associated with epigenetics.Keywords: melanoma, LHPP, nanoparticle, gene therapy, epigenetics

Chunhao Lin,1,2,* Shoumin Bai,1,3,* Tao Du,4 Yiming Lai,2 Xianju Chen,5 Shengmeng Peng,2 Xiaoming Ma,2 Wanhua Wu,2 Zhenghui Guo,1,2 Hai Huang1,2 1Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China; 2Department of Urology, Sun Yatsen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China; 3Department of Radiation Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China; 4Department of Obstetrics and Gynecology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China; 5Department of Urology, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen 518017, China *These authors contributed equally to this work Background: Biological mechanism of prostate cancer (PCa) recurrence and progress is complex but many of the key elements are not fully understood. Polo-like kinases (Plks) represent a family of highly conserved serine–threonine kinases that play essential roles in cell cycle progression. Plk3 plays contradictory roles in different cancers. However, the roles of Plk3 in PCa remain largely unexplored. Methods: Kaplan–Meier analysis and Cox regression analysis were performed to evaluate the relationship between Plk3 and prognosis of patients with PCa. Gene set enrichment analysis (GSEA) was conducted to evaluate proliferation and metastasis gene sets using The Cancer Genome Atlas Dataset. MTS assay, clone formation assay, cell migration, and wound healing assay were carried out to investigate biological functions of Plk3. Results: We found that high Plk3 expression was closely correlated with poor prognosis. GSEA revealed that Plk3 was involved in proliferation and metastasis. Loss-of-function assays demonstrated that Plk3 promoted proliferation and metastasis in PCa cells in vitro. Conclusion: We discovered that Plk3 plays a critical role in PCa, indicating that it may be a potential prognostic marker and help predict the progression, especially recurrence of PCa. Keywords: prostate cancer, polo-like kinase 3, recurrence, proliferation, migration, GSEA

Vascular structures in natural systems are able to provide high mass transport through high surface areas and optimized structure. Few synthetic material fabrication techniques are able to mimic the complexity of these structures while maintaining scalability. The Vaporization of a Sacrificial Component (VaSC) process is able to do so. This process uses sacrificial fibers as a template to form hollow, cylindrical microchannels embedded within a matrix. Tin (II) oxalate (SnOx) is embedded within poly(lactic) acid (PLA) fibers which facilitates the use of this process. The SnOx catalyzes the depolymerization of the PLA fibers at lower temperatures. The lactic acid monomers are gaseous at these temperatures and can be removed from the embedded matrix at temperatures that do not damage the matrix. Here we show a method for aligning these fibers using micromachined plates and a tensioning device to create complex patterns of three-dimensionally arrayed microchannels. The process allows the exploration of virtually any arrangement of fiber topologies and structures.

Tong Zhang,1 Yunjuan Zhao,2 Tao Du,2 Xiaodan Zhang,2 Xiaowei Li,2 Ruike Liu,3 Yunlin Wang,4 Bo Chen,5 Ling He,6 Wangen Li2 1Department of Endocrinology, The Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, People’s Republic of China; 2Department of Endocrinology, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou 510260, People’s Republic of China; 3Department of Endocrinology, The Third People’s Hospital of Dongguan, Dongguan 523326, People’s Republic of China; 4Department of Endocrinology, Foshan Hospital, Southern Medical University, Foshan 528000, People’s Republic of China; 5Department of Endocrinology, Guangdong Second Provincial General Hospital, Guangzhou 510317, People’s Republic of China; 6Department of Endocrinology, Guangzhou First People’s Hospital, Guangzhou 510180, People’s Republic of China Background: Patient-centered care is respectful to a patient’s preference. All prior clinical trials on patient self-titration algorithms for basal insulin were decided by physicians. We hypothesized that patients and physicians have different preferences. Patients and methods: Physicians and diabetes patients were asked to choose their preferred insulin glargine self-titration algorithm among 5 algorithms. Algorithm 1, 1 U increase once daily; algorithm 2, 2 U increase every 3 days; algorithm 3, 3 U increase every 3 days; algorithm 4, titration every 3 days according to fasting blood glucose, and algorithm 5, weekly titration 2–8 U based on 3-day mean fasting blood glucose levels. Results: Eleven (5.2%) out of 210 physicians and 180 (90.9%) out of 198 patients preferred algorithm 1 (χ2=300.4, p=0.000). In contrast, 195 (92.9%) physicians and 18 (9.1%) patients preferred algorithm 2 (χ2=286.6, p=0.000). In addition, 4 (1.9%) physicians but no patients preferred algorithm 3 (χ2=2.099, p=0.124). Neither physicians nor patients chose algorithms 4 or 5. Most physicians preferred algorithm 2 since it is recommended by guidelines, but most patients preferred algorithm 1 for its simplicity. Conclusion: Patients had different preferences compared with physicians. Attention should be given to patients’ preferences to increase adherence and improve glycemic control. Keywords: type 2 diabetes, titration algorithm, insulin glargine, basal insulin, self-adjustment, shared decision-making, adherence

Tianyi Kang,1 Chenyang Li,1,2 Ting Du,1 Yujiao Wu,1 Yuping Yang,1 Xuan Liu,1 Qianqian Zhang,1 Xiaoping Xu,3 Maling Gou1,4 1State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, People’s Republic of China; 2Shanghai Institutes for Biological Science, Chinese Academy of Science, Shanghai 200031, People’s Republic of China; 3West China School of Pharmacy, Sichuan University, Chengdu 610041, People’s Republic of China; 4Co-Innovation Center for Micro/Nano Optoelectronic Materials and Devices, Chongqing 402160, People’s Republic of China Background: Melittin, the main active peptide ingredient of bee venom, can cause severe cell membrane lysis due to its robust interaction with negatively charged phospholipids. So far, no effective anti-melittin vaccine has been developed to protect people from undesired melittin intoxication.Methods: Herein, we prepared a polydiacetylene (PDA) nanoparticle with cell membrane-mimic surface to complex melittin, forming an anti-melittin vaccine (PDA–melittin).Results: PDA nanoparticles could effectively combine with melittin and neutralize its toxicity. PDA–melittin nanocomplex is demonstrated to enhance melittin uptake by DCs and stimulate strong melittin-specific immunity. Mice immunized with PDA–melittin nanocomplex showed higher survival rate after exposion to melittin than untreated mice.Conclusion: The PDA–melittin nanocomplex can efficiently and safely generate a specific immunity against melittin to protect body from melittin intoxication, providing a new method with potential clinical application for the treatment of melittin intoxication.Keywords: PDA nanoparticles, melittin, toxoid vaccine, immunity