Publications by authors named "Qingling Li"

84 Publications

Serum Lysophosphatidic Acid Measurement by Liquid Chromatography-Mass Spectrometry in COPD Patients.

J Am Soc Mass Spectrom 2021 Mar 23. Epub 2021 Mar 23.

Lysophospholipids are bioactive signaling molecules derived from cell membrane glycerophospholipids or sphingolipids and are highly regulated under normal physiological conditions. Lysophosphatidic acids (LPAs) are a class of lysophospholipids that act on G-protein-coupled receptors to exert a variety of cellular functions. Dysregulation of phospholipase activity and consequently LPA synthesis in serum have been linked to inflammation, such as seen in chronic obstructive pulmonary disease (COPD). The accurate measurement of phospholipids is critical for evaluating their dysregulation in disease. In this study, we optimized experimental parameters for the sensitive measurement of LPAs. We validated the method based on matrix, linearity, accuracy, precision, and stability. An investigation into sample extraction processes emphasized that the common practice of including low concentration of hydrochloric acid in the extraction buffer causes an overestimation of lipid recovery. The liquid chromatography gradient was optimized to separate various lysophospholipid classes. After optimization, detection limits of LPA were sufficiently sensitive for subsequent analysis, ranging from 2 to 8 nM. The validated workflow was applied to a cohort of healthy donor and COPD patient sera. Eight LPA species were identified, and five unique species of LPA were quantified. Most LPA species increased significantly in COPD patients compared to healthy donors. The correlation between LPAs and other demographic parameters was further investigated in a sample set of over 200 baseline patient sera from a COPD clinical trial. For the first time, LPAs other than the two most abundant and readily detectable moieties are quantified in COPD patients using validated methods, opening the door to downstream biomarker evaluation in respiratory disease.
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http://dx.doi.org/10.1021/jasms.0c00429DOI Listing
March 2021

Ocular phenotypes in a mouse model of impaired glucocerebrosidase activity.

Sci Rep 2021 Mar 16;11(1):6079. Epub 2021 Mar 16.

Departments of Neuroscience, Genentech Inc., 1 DNA Way, South San Francisco, CA, 94080, USA.

Mutations in the GBA1 gene encoding glucocerebrosidase (GCase) are linked to Gaucher (GD) and Parkinson's Disease (PD). Since some GD and PD patients develop ocular phenotypes, we determined whether ocular phenotypes might result from impaired GCase activity and the corresponding accumulation of glucosylceramide (GluCer) and glucosylsphingosine (GluSph) in the Gba1 knock-in (Gba KI/KI; "KI") mouse. Gba KI mice developed age-dependent pupil dilation deficits to an anti-muscarinic agent; histologically, the iris covered the anterior part of the lens with adhesions between the iris and the anterior surface of the lens (posterior synechia). This may prevent pupil dilation in general, beyond an un-responsiveness of the iris to anti-muscarinics. Gba KI mice displayed atrophy and pigment dispersion of the iris, and occlusion of the iridocorneal angle by pigment-laden cells, reminiscent of secondary open angle glaucoma. Gba KI mice showed progressive thinning of the retina consistent with retinal degeneration. GluSph levels were increased in the anterior and posterior segments of the eye, suggesting that accumulation of lipids in the eye may contribute to degeneration in this compartment. We conclude that the Gba KI model provides robust and reproducible eye phenotypes which may be used to test for efficacy and establish biomarkers for GBA1-related therapies.
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http://dx.doi.org/10.1038/s41598-021-85528-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7971029PMC
March 2021

Nanomedicine promotes ferroptosis to inhibit tumour proliferation in vivo.

Redox Biol 2021 Feb 20:101908. Epub 2021 Feb 20.

Department of General Surgery, Geriatrics, Obstetrics and Gynecology, Division of Pulmonary and Critical Care Medicine, Institute of Precision Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China. Electronic address:

miR-101-3p may play a therapeutic role in various tumours. However, its anti-tumour mechanism remains unclear, and a definitive strategy to treat tumour cells in vivo is lacking. The objective of this study was to investigate the inhibitory mechanism of miR-101-3p on tumour cells and to develop relevant nanomedicines for in vivo therapy. The expression levels of miR-101-3p and its target protein TBLR1 in tumour tissues and cells were detected, and their relationship with ferroptosis was clarified. Furthermore, the efficacy of nanocarriers in achieving in vivo therapeutic gene delivery was evaluated. Nanomedicine was further developed, with the anti-proliferative in vivo therapeutic effect validated using a subcutaneous xenograft cancer model. The expression level of miR-101-3p negatively correlated with clinical tumour size and TNM stage. miR-101-3p restores ferroptosis in tumour cells by directly targeting TBLR1, which in turn promotes apoptosis and inhibits proliferation. We developed nanomedicine that can deliver miR-101-3p to tumour cells in vivo to achieve ferroptosis recovery, as well as to inhibit in vivo tumour proliferation. The miR-101-3p/TBLR1 axis plays an important role in tumour ferroptosis. Nanopharmaceuticals that increase miR-101-3p levels may be effective therapies to inhibit tumour proliferation.
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http://dx.doi.org/10.1016/j.redox.2021.101908DOI Listing
February 2021

Moxibustion Improves Chronic Heart Failure by Inhibiting Autophagy and Inflammation via Upregulation of mTOR Expression.

Evid Based Complement Alternat Med 2021 2;2021:6635876. Epub 2021 Feb 2.

School of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.

How moxibustion improves chronic heart failure is extremely complex and still unclear. This study aimed to explore whether moxibustion inhibits autophagy and reduces inflammation by regulating mTOR expression to induce myocardial protective effects and alleviate symptoms associated with chronic heart failure. Echocardiography was used to detect cardiac function and cardiac structure of rats, including heart rate (HR), left atrium diameter (LA), left ventricular diameter (LV), left ventricular posterior wall (LVPW), interventricular septum (IVS), ejection fraction (EF), and fractional shortening (FS). BNP and NT-pro BNP levels were measured by enzyme-linked immunosorbent assay (ELISA). Autophagy-associated protein (ATG) genes and mTOR were detected by PCR. The expression of mTOR and phosphorylated-mTOR was detected through western blotting of proteins from myocardial tissue samples. The left ventricular inflammatory response was detected by immunohistochemistry and included ICAM-1, VCAM-1, MMP-2, and MMP-9 expression. The relationship between autophagy and inflammation was analyzed by correlation analysis. The results from echocardiography and ELISA showed that moxibustion could significantly improve heart function and structure. Western blot and PCR results showed that moxibustion treatment elevated mTOR expression. Further, moxibustion could inhibit autophagy and regulate the expression of key autophagy-related genes, including Vps34, ATG3, ATG5, ATG7, ATG12, and ATG13. By contrast, rapamycin could partially reduce the effects of moxibustion. Immunohistochemistry results indicated that moxibustion could reduce myocardial inflammation. Moreover, there was a positive correlation between autophagy and inflammation. Moxibustion can protect cardiac function in rats with heart failure, possibly inhibiting excessive autophagy of cardiomyocytes and reducing inflammatory reactions through the elevation of mTOR expression.
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http://dx.doi.org/10.1155/2021/6635876DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7872756PMC
February 2021

NINJ1 mediates plasma membrane rupture during lytic cell death.

Nature 2021 Mar 20;591(7848):131-136. Epub 2021 Jan 20.

Department of Physiological Chemistry, Genentech Inc., South San Francisco, CA, USA.

Plasma membrane rupture (PMR) is the final cataclysmic event in lytic cell death. PMR releases intracellular molecules known as damage-associated molecular patterns (DAMPs) that propagate the inflammatory response. The underlying mechanism of PMR, however, is unknown. Here we show that the cell-surface NINJ1 protein, which contains two transmembrane regions, has an essential role in the induction of PMR. A forward-genetic screen of randomly mutagenized mice linked NINJ1 to PMR. Ninj1 macrophages exhibited impaired PMR in response to diverse inducers of pyroptotic, necrotic and apoptotic cell death, and were unable to release numerous intracellular proteins including HMGB1 (a known DAMP) and LDH (a standard measure of PMR). Ninj1 macrophages died, but with a distinctive and persistent ballooned morphology, attributable to defective disintegration of bubble-like herniations. Ninj1 mice were more susceptible than wild-type mice to infection with Citrobacter rodentium, which suggests a role for PMR in anti-bacterial host defence. Mechanistically, NINJ1 used an evolutionarily conserved extracellular domain for oligomerization and subsequent PMR. The discovery of NINJ1 as a mediator of PMR overturns the long-held idea that cell death-related PMR is a passive event.
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http://dx.doi.org/10.1038/s41586-021-03218-7DOI Listing
March 2021

Application of a microfluidic paper-based bioimmunosensor with laser-induced fluorescence detection in the determination of alpha-fetoprotein from serum of hepatopaths.

Talanta 2021 Jan 14;221:121660. Epub 2020 Sep 14.

Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Research Institute of Photocatalysis, School of Chemistry, Fuzhou University, Fuzhou, 350116, PR China.

As a common plasma protein, alpha-fetoprotein (AFP) is widely applied as the tumor biomarker for the diagnosis of many cancers. To develop a low cost, high sensitive and high-throughput method for the determination of AFP is significant for the disease diagnosis. In this work, an immunoassay with sandwich-type structures was performed on a paper-based chip for the analysis of AFP. AFP could be captured by the primary antibodies which were immobilized on the paper by chitosan. On the secondary antibodies, the modified initiator DNAs could trigger the hybridization chain reaction to amplify the fluorescence signals for AFP. A laser-induced fluorescence detector coupled with an interface was applied to detect the targets on the paper-based chip. Under the optimized conditions, the detection limit for AFP was 1.0 pg/mL. For every test, the sample solution consumption only was 10 μL. Finally, the method was applied to determine the AFP in serum of normal person and hepatopaths with hepatic malignant tumor, chronic hepatitis B and other suspected liver diseases. The AFP could be found from all of the samples and the results were similar to that obtained by chemiluminescence immunoassay. The recoveries for AFP ranged from 93.8% to 106%, which indicated the method was reliable. The method based on paper chip had great potential in the application of AFP determination.
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http://dx.doi.org/10.1016/j.talanta.2020.121660DOI Listing
January 2021

Mammary synchronous mucinous cystadenocarcinoma and columnar cell mucinous carcinoma: a case report.

Int J Clin Exp Pathol 2020 1;13(9):2381-2386. Epub 2020 Sep 1.

Department of Pathology, Xiangya Hospital, Central South University Changsha, Hunan, China.

Mucin-producing carcinomas are unusual primary malignancies of breast, and constitute about 1-4 percent of total breast cancer. The mammary mucin producing carcinomas are divided into 4 histologic subtypes according to WHO classification, including mucinous carcinoma, mucinous cystadenocarcinoma (MCA), columnar cell mucinous carcinoma (CCMC), and signet ring cell carcinoma. However, the synchronous primary MCA and CCMC of breast is a very rare case presentation. The case reported a 56-year-old female, who presented with right mammary lumps and nipple discharge about 1 year. Imaging examinations revealed multiple cystic and solid nodules in upper outer quadrant of right breast, associated with ectatic ducts. Serum levels of tumor markers were normal. Right mammary lumpectomy revealed mucinous carcinoma, modified radical mastectomy, and lymph node dissection were carried out. For neoplastic cells, ER and PR were positive, HER2 (1+) was negative, Ki67 was low expression (3-5%). There was no metastatic carcinoma in lymph nodes (0/8). Modified radical mastectomy and lymph node dissections were carried out. Tamoxifen was chosen for adjuvant therapy. After a 3 month follow up, the patient survived without recurrences and distant metastasis. We report the first synchronous primary MCA and CCMC of breast with molecular subtype of Luminal A.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7539871PMC
September 2020

Structure of the essential inner membrane lipopolysaccharide-PbgA complex.

Nature 2020 08 12;584(7821):479-483. Epub 2020 Aug 12.

Infectious Diseases, Genentech Inc., South San Francisco, CA, USA.

Lipopolysaccharide (LPS) resides in the outer membrane of Gram-negative bacteria where it is responsible for barrier function. LPS can cause death as a result of septic shock, and its lipid A core is the target of polymyxin antibiotics. Despite the clinical importance of polymyxins and the emergence of multidrug resistant strains, our understanding of the bacterial factors that regulate LPS biogenesis is incomplete. Here we characterize the inner membrane protein PbgA and report that its depletion attenuates the virulence of Escherichia coli by reducing levels of LPS and outer membrane integrity. In contrast to previous claims that PbgA functions as a cardiolipin transporter, our structural analyses and physiological studies identify a lipid A-binding motif along the periplasmic leaflet of the inner membrane. Synthetic PbgA-derived peptides selectively bind to LPS in vitro and inhibit the growth of diverse Gram-negative bacteria, including polymyxin-resistant strains. Proteomic, genetic and pharmacological experiments uncover a model in which direct periplasmic sensing of LPS by PbgA coordinates the biosynthesis of lipid A by regulating the stability of LpxC, a key cytoplasmic biosynthetic enzyme. In summary, we find that PbgA has an unexpected but essential role in the regulation of LPS biogenesis, presents a new structural basis for the selective recognition of lipids, and provides opportunities for future antibiotic discovery.
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http://dx.doi.org/10.1038/s41586-020-2597-xDOI Listing
August 2020

Optimization of process parameters for preparation of polystyrene PM2.5 particles by supercritical antisolvent method using BBD-RSM.

Sci Rep 2020 Jul 7;10(1):11187. Epub 2020 Jul 7.

School of Electromechanical Engineering, Qingdao University of Science and Technology, Qingdao, 266061, China.

The objective of this study is to optimize the process parameters for preparing polystyrene (PS) PM2.5 particles by supercritical antisolvent (SAS) method. Toluene was selected as the solvent and supercritical carbon dioxide (SC-CO) was used as the antisolvent. The Box-Behnken design-response surface method was applied to investigate the effect of crystallizer pressure, PS massic concentration, flow ratio of CO/solution and crystallizer temperature on the size and the distribution of PS particles, systematically. It is found that crystallizer temperature is the most significant variable on the size and the distribution of PS particles, followed by flow ratio of CO/solution and PS massic concentration, and crystallizer pressure is the slightest significant factor. The particle size increases with the increase of crystallizer temperature. The optimum conditions are obtained as crystallizer pressure 9.8 MPa, PS massic concentration 1.6 wt%, flow ratio of CO/solution 140 g/g and crystallizer temperature 309 K. Under these conditions, the PS particle with the size of 2.78 μm and a narrow size distribution has been prepared, meeting PM2.5 standard aerosols. The results suggest that it is feasible to produce PM2.5 standard aerosols by SAS.
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http://dx.doi.org/10.1038/s41598-020-67994-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7341738PMC
July 2020

Long non-coding RNA TUSC8 inhibits breast cancer growth and metastasis via miR-190b-5p/MYLIP axis.

Aging (Albany NY) 2020 02 9;12(3):2974-2991. Epub 2020 Feb 9.

Department of Radiology, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China.

The lncRNA tumor suppressor candidate 8 (TUSC8) plays a critical role in the development of several cancers. However, the biological functions and underlying molecular mechanisms of TUSC8 with respect to breast cancer remain largely unclear. Here, we found that TUSC8 was significantly down-regulated in breast cancer tissues and its high expression predicted better prognosis of breast cancer patients. Functionally, knock-down of TUSC8 drastically promoted the proliferation, migration and invasion of breast cancer cells and facilitated tumorigenicity and metastasis . Mechanistically, the results of luciferase reporter, RIP and RNA pull-down assays proved that TUSC8 functioned as molecular sponge for miR-190b-5p. Furthermore, we showed that TUSC8 served as a competing endogenous RNA (ceRNA) of myosin regulatory light chain interacting protein (MYLIP) through competitively binding with miR-190b-5p and suppressed breast cancer metastasis through regulating the expression of epithelial-mesenchymal transition (EMT) related markers. Clinically, the receiver operating characteristic curve (ROC) analyses revealed that the combination usage of TUSC8 and MYLIP might become novel promising diagnostic biomarkers for breast cancer. Taken together, these results suggested that TUSC8 inhibited breast cancer growth and metastasis via miR-190b-5p/MYLIP axis, providing us new insights into developing potential therapeutic targets for breast cancer patients.
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http://dx.doi.org/10.18632/aging.102791DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041739PMC
February 2020

Three-Dimensional Superlithiophilic Interphase for Dendrite-Free Lithium Metal Anodes.

ACS Appl Mater Interfaces 2020 Feb 22;12(5):5767-5774. Epub 2020 Jan 22.

School of Chemistry and Chemical Engineering , Henan Normal University , Xinxiang 453007 , China.

Lithium metal is among the most promising anode candidates of high-energy-density batteries. However, the formed dendrites result in low Coulombic efficiency and serious security issues. Designing lithiophilic sites is one of the effective strategies to control Li deposition. Herein, we propose a three-dimensional lithiophilic N-rich carbon nanofiber with the decoration of ZnO granules as a protective layer for a dendrite-free lithium metal anode. Theoretical evaluation indicates the synergistic effects of lithiophilic ZnO and N-containing functional groups enhance lithium adsorption and trigger uniform deposition. With the lithiophilic interlayer, the lithium deposition overpotential is only ∼20, 50, and 74 mV at 1, 3, and 6 mA cm, respectively, which are much lower than those without the functional interlayer (∼55, 130, and 238 mV). The average Coulombic efficiency of lithium stripping and plating is up to ∼97.4% (94.0% for that without the interlayer) at 0.5 mA cm. Meanwhile, the Li|LiFePO full cell with the superlithiophilic interlayer demonstrates a high capacity retention rate of 99.6% (91.0% for that without the interlayer) over 200 cycles at 1 C. The introduction of the lithiophilic interphase could provide a convenient strategy and guidance to design the configuration for the practical application of Li metal batteries.
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http://dx.doi.org/10.1021/acsami.9b18315DOI Listing
February 2020

Efficacy of a New Therapeutic Option for Vulvar Intraepithelial Neoplasia: Superficial Shaving Combined With Photodynamic Therapy.

Lasers Surg Med 2020 07 10;52(6):488-495. Epub 2019 Nov 10.

Department of Dermatology, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, 410008, Hunan, China.

Background And Objective: Vulvar intraepithelial neoplasia (VIN), a precancerous lesion, is difficult to treat by excision or ablation due to high recurrence rates. Photodynamic therapy (PDT) is a minimally invasive therapeutic procedure and is now widely used to treat non-melanoma skin diseases. However, the clinical response rates of VIN to single PDT are unstable. The reason may be the limited light penetration into deep tissues.

Objective: To retrospectively evaluate the clinical response and recurrence of VIN after combined treatment with superficial shaving and PDT.

Study Design/materials And Methods: Seventeen patients with VIN were enrolled. All patients had multifocal high-grade VIN that had failed to respond to various therapies. Superficial shaving was performed only once and prior to the first 5-aminolaevulinic acid (5-ALA)-PDT cycle. Generally, the procedure of 5-ALA PDT for each patient was performed in three sessions. Clinical response, recurrence, cosmetic outcomes, adverse events, patient satisfaction, quality of life, and mental health were assessed. The expression of p16 and Ki-67 in pre- and post-treatment tissue was detected.

Results: A clinical response of 94% was observed in 17 patients, who were administered combination therapy, over an observation period of 12 months. Approximately, 71% of patients had excellent cosmetic outcomes. All patients had satisfactory therapeutic effects and significant improvements in quality of life and mental health. Downregulation of p16 and Ki-67 may have been correlated with recurrence after 5-ALA-PDT.

Conclusion: Combined treatment with superficial shaving and 5-ALA-PDT is a safe and effective option for VIN. In particular, combination therapy is recommended for patients with large, multifocal, high-grade lesions; repeated recurrence; and strong willingness to maintain vulvar configuration and function. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.
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http://dx.doi.org/10.1002/lsm.23185DOI Listing
July 2020

Biological actions of Cu/Zn coimplanted TiN on Ti-6Al-4V alloy.

Biointerphases 2019 10 15;14(5):051008. Epub 2019 Oct 15.

College of Physics and Materials Science, Tianjin Normal University, Tianjin 300387, China.

Ti-6Al-4V alloy, as a widely used orthopedic and dental implant, has excellent biocompatibility and machinability. However, its poor corrosion resistance and antibacterial property may lead to tissue inflammation and postoperative infection, which hinders its further development. In this paper, to solve the above problems, copper (Cu) and zinc (Zn) ions were coimplanted into a titanium nitride (TiN) coated Ti-6Al-4V alloy via a plasma immersion ion implantation system (PIII). Then, the structure and composition of Cu/Zn coimplanted TiN (Cu/Zn-TiN-PIII) were characterized by scanning electron microscopy and x-ray photoelectron spectroscopy. Also, the results of the corrosion test, the water contact angles test, and the protein electrophoresis experiment showed that the corrosion resistance, hydrophilicity, and the protein adsorption capacity of Cu/Zn-TiN-PIII were improved simultaneously. In addition, compared with TiN-PIII, Cu/Zn-TiN-PIII promoted both cytocompatibility and the antibacterial property according to L929 cells and Escherichia coli assays in vitro. Therefore, Cu/Zn-TiN-PIII may be a good candidate for orthopedic implants.
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http://dx.doi.org/10.1116/1.5116640DOI Listing
October 2019

The RIPK4-IRF6 signalling axis safeguards epidermal differentiation and barrier function.

Nature 2019 10 2;574(7777):249-253. Epub 2019 Oct 2.

Department of Physiological Chemistry, Genentech, South San Francisco, CA, USA.

The integrity of the mammalian epidermis depends on a balance of proliferation and differentiation in the resident population of stem cells. The kinase RIPK4 and the transcription factor IRF6 are mutated in severe developmental syndromes in humans, and mice lacking these genes display epidermal hyperproliferation and soft-tissue fusions that result in neonatal lethality. Our understanding of how these genes control epidermal differentiation is incomplete. Here we show that the role of RIPK4 in mouse development requires its kinase activity; that RIPK4 and IRF6 expressed in the epidermis regulate the same biological processes; and that the phosphorylation of IRF6 at Ser413 and Ser424 primes IRF6 for activation. Using RNA sequencing (RNA-seq), histone chromatin immunoprecipitation followed by sequencing (ChIP-seq) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) of skin in wild-type and IRF6-deficient mouse embryos, we define the transcriptional programs that are regulated by IRF6 during epidermal differentiation. IRF6 was enriched at bivalent promoters, and IRF6 deficiency caused defective expression of genes that are involved in the metabolism of lipids and the formation of tight junctions. Accordingly, the lipid composition of the stratum corneum of Irf6 skin was abnormal, culminating in a severe defect in the function of the epidermal barrier. Collectively, our results explain how RIPK4 and IRF6 function to ensure the integrity of the epidermis and provide mechanistic insights into why developmental syndromes that are characterized by orofacial, skin and genital abnormalities result when this axis goes awry.
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http://dx.doi.org/10.1038/s41586-019-1615-3DOI Listing
October 2019

Consecutive Sorting and Phenotypic Counting of CTCs by an Optofluidic Flow Cytometer.

Anal Chem 2019 11 14;91(21):14133-14140. Epub 2019 Oct 14.

College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Institute of Molecular and Nano Science , Shandong Normal University , Jinan 250014 , P. R. China.

Circulating tumor cell (CTC) analysis has been approved for cancer diagnosis and monitoring. However, efficient sorting and high-through phenotypic counting of CTCs from peripheral blood is still a challenge. In this manuscript, we propose an optofluidic flow cytometer (OFCM), which integrates a multistage microfluidic chip and a four-color fluorescence detection system. The OFCM can automatically complete CTC separation, 3D focusing in the microchannel, single-cell phenotypic analysis, and counting at 1.2 mL of whole blood/hour. A high recovery greater than 95% was obtained. Using the OFCM, we analyzed the epithelial-to-mesenchymal transition (EMT) phenotype of CTCs in patients with breast cancer and patients with nonsmall cell lung cancer, which proved that the OFCM is adaptable for phenotypic counting of various CTCs based on the fluorescence labeling of varied biomarkers. We believe that this OFCM will provide a convenient and efficient device for clinical liquid biopsy of tumors.
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http://dx.doi.org/10.1021/acs.analchem.9b04035DOI Listing
November 2019

High-throughput and ultra-sensitive single-cell profiling of multiple microRNAs and identification of human cancer.

Chem Commun (Camb) 2019 Aug;55(70):10404-10407

College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging, Key Laboratory of Molecular and Nano Probes, Ministry of Education Shandong Normal University, Jinan 250014, Shandong, P. R. China.

We established an efficient method for single-cell miRNA analysis by droplet microfluidics, which has high sensitivity of single molecule detection and high throughput. Single-cell analysis of multiple miRNAs in various cells shows that miRNA expression is closely related to cancer type. CTC analysis shows that the method is applicable for rare cell analysis.
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http://dx.doi.org/10.1039/c9cc05553cDOI Listing
August 2019

Downregulation of miR‑98 contributes to hypoxic pulmonary hypertension by targeting ALK1.

Mol Med Rep 2019 Sep 9;20(3):2167-2176. Epub 2019 Jul 9.

Department of Respiratory Medicine, Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University, Xuzhou, Jiangsu 221002, P.R. China.

Chronic hypoxia is one of the most common causes of secondary pulmonary hypertension, the mechanisms of which remain unclear. MicroRNAs (miRNAs) are small, noncoding RNAs that inhibit the translation or accelerate the degradation of mRNA. Previous studies have demonstrated that deregulated miRNA expression contributes to various cellular processes including cell apoptosis and proliferation, which are mediated by hypoxia. In the present study, the expression of miR‑98 was identified to be decreased in the lung tissue of a hypoxic pulmonary hypertension (HPH) rat model and pulmonary artery (PA) smooth muscle cells (PASMCs), which was induced by hypoxia. By transfecting miR‑98 mimics into PASMCs, the high expression of miR‑98 inhibited cell proliferation, but upregulated hypoxia‑induced PASMCs apoptosis. However, these effects of miR‑98 mimics on PASMCs were reversed by ALK1 (activin receptor‑like kinase‑1) overexpression. ALK1 was identified as a candidate target of miR‑98. In addition, overexpressing miR‑98 markedly decreased the pulmonary artery wall thickness and the right ventricular systolic pressure in rats induced by hypoxia. These results provided clear evidence that miR‑98 was a direct regulator of ALK1, and that the downregulation of miR‑98 contributed to the pathogenesis of HPH. These results provide a novel potential therapeutic strategy for the treatment of HPH.
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http://dx.doi.org/10.3892/mmr.2019.10482DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6691262PMC
September 2019

MRI-Based Radiomics Predicts Tumor Response to Neoadjuvant Chemoradiotherapy in Locally Advanced Rectal Cancer.

Front Oncol 2019 26;9:552. Epub 2019 Jun 26.

Department of Diagnostic Radiology, City of Hope National Medical Center, Duarte, CA, United States.

Conventional methods for predicting treatment response to neoadjuvant chemoradiotherapy (nCRT) in patients with locally advanced rectal cancer (LARC) are limited. This study retrospectively recruited 134 LARC patients who underwent standard nCRT followed by total mesorectal excision surgery in our institution. Based on pre-operative axial T2-weighted images, machine learning radiomics was performed. A receiver operating characteristic (ROC) curve was performed to test the efficiencies of the predictive model. Among the 134 patients, 32 (23.9%) achieved pathological complete response (pCR), 69 (51.5%) achieved a good response, and 91 (67.9%) achieved down-staging. For prediction of pCR, good-response, and down-staging, the predictive model demonstrated high classification efficiencies, with an AUC value of 0.91 (95% CI: 0.83-0.98), 0.90 (95% CI: 0.83-0.97), and 0.93 (95% CI: 0.87-0.98), respectively. Our machine learning radiomics model showed promise for predicting response to nCRT in patients with LARC. Our predictive model based on the commonly used T2-weighted images on pelvic Magnetic Resonance Imaging (MRI) scans has the potential to be adapted in clinical practice. Methods for predicting the response of the locally advanced rectal cancer (LARC, T3-4, or N+) to neoadjuvant chemoradiotherapy (nCRT) is lacking. In the present study, we developed a new machine learning radiomics method based on T2-weighted images. As a non-invasive tool, this method facilitates prediction performance effectively. It achieves a satisfactory overall diagnostic accuracy for predicting of pCR, good response, and down-staging show an AUC of 0.908, 0.902, and 0.930 in LARC patients, respectively.
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http://dx.doi.org/10.3389/fonc.2019.00552DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6606732PMC
June 2019

Effects of Zn and Ag Ratio on Cell Adhesion and Antibacterial Properties of Zn/Ag Coimplanted TiN.

ACS Biomater Sci Eng 2019 Jul 24;5(7):3303-3310. Epub 2019 Jun 24.

College of Physics and Materials Science and Tianjin International Joint Research Center of Surface Technology for Energy Storage Materials, Tianjin Normal University, Tianjin 300387, China.

Titanium alloy (Ti-6Al-4 V) has good biocompatibility and mechanical properties, but its biologically inert and poor antibacterial property limit its application in the field of medical implants. In this study, titanium nitride (TiN) is first deposited on the surface of titanium alloy to wrap toxic elements (for example, vanadium). Then Zn and Ag ions with different ratios are simultaneously implanted into TiN via a plasma immersion ion implantation (PIII) system. X-ray photoelectron spectrometer proves the presence of Zn, Ag, and TiN on the surface of Zn/Ag-TiN. Protein adsorption suggests that, with the increase of Zn ratio, the adsorption ability is enhanced. Biological assays indicate that Zn/Ag(1/0.3)-TiN has the best cell proliferation and adhesion, because it has a larger ratio of Zn. Zn/Ag(1/2)-TiN has the best antibacterial effect and followed by Zn/Ag(1/0.3)-TiN, since Ag plays a major role in antibacterial activity and Zn is auxiliary. But a large ratio of Ag not only impairs cells but also has a negative effect on antibacterial activity. In conclusion, Zn/Ag(1/0.3)-TiN offers both excellent cytocompatibility and antibacterial activity and has great potential in orthopedic implants.
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http://dx.doi.org/10.1021/acsbiomaterials.9b00248DOI Listing
July 2019

A Stromal Niche Defined by Expression of the Transcription Factor WT1 Mediates Programming and Homeostasis of Cavity-Resident Macrophages.

Immunity 2019 07 20;51(1):119-130.e5. Epub 2019 Jun 20.

Department of Cancer Immunology, Genentech, South San Francisco, CA 94080, USA. Electronic address:

Tissue-resident macrophages require specific milieus for the maintenance of defining gene-expression programs. Expression of the transcription factor GATA6 is required for the homeostasis, function and localization of peritoneal cavity-resident macrophages. Gata6 expression is maintained in a non-cell autonomous manner and is elicited by the vitamin A metabolite, retinoic acid. Here, we found that the GATA6 transcriptional program is a common feature of macrophages residing in all visceral body cavities. Retinoic acid-dependent and -independent hallmark genes of GATA6 macrophages were induced by mesothelial and fibroblastic stromal cells that express the transcription factor Wilms' Tumor 1 (WT1), which drives the expression of two rate-limiting enzymes in retinol metabolism. Depletion of Wt1 stromal cells reduced the frequency of GATA6 macrophages in the peritoneal, pleural and pericardial cavities. Thus, Wt1 mesothelial and fibroblastic stromal cells constitute essential niche components supporting the tissue-specifying transcriptional landscape and homeostasis of cavity-resident macrophages.
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http://dx.doi.org/10.1016/j.immuni.2019.05.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6814267PMC
July 2019

Development of a Non-Coding-RNA-based EMT/CSC Inhibitory Nanomedicine for In Vivo Treatment and Monitoring of HCC.

Adv Sci (Weinh) 2019 May 7;6(9):1801885. Epub 2019 Mar 7.

Department of General Surgery The First Affiliated Hospital of Sun Yat-Sen University Guangzhou 510080 China.

The objective of this study is to improve the overall prognosis of patients with hepatocellular carcinoma (HCC); therefore, new therapeutic methods that can be used in vivo are urgently needed. In this study, the relationship between the quantities of microRNA (miR)-125b-5p in clinical specimens and clinicopathological parameters is analyzed. A folate-conjugated nanocarrier is used to transfect miR-125b-5p in vivo and to observe the therapeutic effect on HCC. The inhibitory effect and mechanism of miR-125b-5p on hepatoma cells are also studied. Data from clinical specimens and in vitro experiments confirm that the miR-125b-5p quantity is negatively correlated with progression, and the target protein that regulates the epithelial-mesenchymal transition (EMT)/cancer stem cells (CSC) potential in HCC is STAT3. The miR-125b-5p/STAT3 axis inhibits the invasion, migration, and growth of HCC via inactivation of the wnt/β-Catenin pathway. miR-125b-5p-loaded nanomedicine effectively inhibits the EMT/CSC potential of hepatoma cells in vivo together with their magnetic resonance imaging (MRI) visualization characteristics. An HCC-therapeutic and MRI-visible nanomedicine platform that achieves noninvasive treatment effect monitoring and timely individualized treatment course adjustment is developed.
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http://dx.doi.org/10.1002/advs.201801885DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6498119PMC
May 2019

Therapeutic resistance and susceptibility is shaped by cooperative multi-compartment tumor adaptation.

Cell Death Differ 2019 Nov 1;26(11):2416-2429. Epub 2019 Mar 1.

Department of Translational Oncology, Genentech, Inc., 1 DNA Way, South San Francisco, CA, 94080, USA.

Emerging research suggests that multiple tumor compartments can influence treatment responsiveness and relapse, yet the search for therapeutic resistance mechanisms remains largely focused on acquired genomic alterations in cancer cells. Here we show how treatment-induced changes occur in multiple tumor compartments during tumor relapse and can reduce benefit of follow-on therapies. By using serial biopsies, next-generation sequencing, and single-cell transcriptomics, we tracked the evolution of multiple cellular compartments within individual lesions during first-line treatment response, relapse, and second-line therapeutic interventions in an autochthonous model of melanoma. We discovered that although treatment-relapsed tumors remained genetically stable, they converged on a shared resistance phenotype characterized by dramatic changes in tumor cell differentiation state, immune infiltration, and extracellular matrix (ECM) composition. Similar alterations in tumor cell differentiation were also observed in more than half of our treatment-relapsed patient tumors. Tumor cell-state changes were coincident with ECM remodeling and increased tumor stiffness, which alone was sufficient to alter tumor cell fate and reduce treatment responses in melanoma cell lines in vitro. Despite the absence of acquired mutations in the targeted pathway, resistant tumors showed significantly decreased responsiveness to second-line therapy intervention within the same pathway. The ability to preclinically model relapse and refractory settings-while capturing dynamics within and crosstalk between all relevant tumor compartments-provides a unique opportunity to better design and sequence appropriate clinical interventions.
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http://dx.doi.org/10.1038/s41418-019-0310-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6889278PMC
November 2019

Dynamic fluorescent imaging analysis of mitochondrial redox in single cells with a microfluidic device.

Biosens Bioelectron 2019 Mar 11;129:132-138. Epub 2019 Jan 11.

College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Institute of Molecular and Nano Science, Shandong Normal University, Jinan 250014, PR China. Electronic address:

The redox balance in cellular mitochondria is closely related to the physiological and pathological processes of the body. When exposed to external stimuli, the redox state in cells changes dynamically, and presents cell heterogeneity, which creates a need for techniques that can make dynamic and reversible visual analysis of redox in mitochondria at single-cell level. Here we describe a method for single-cell redox analysis based on a microfluidic device combing with a reversible fluorescent probe (Cy-O-ebselen), that enables online culture, labelling and dynamic fluorescent imaging analysis of mitochondrial redox (HO/GSH) change. Using this method, we further explored the dynamic changes of mitochondrial redox state after thermal stimulation or combined thermal-drug stimulation, and analysed the heterogeneous response of cells to external stimuli at the single cell level.
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http://dx.doi.org/10.1016/j.bios.2019.01.005DOI Listing
March 2019

C-reactive protein (CRP) rs3093059C predicts poor mizolastine response in chronic spontaneous urticaria patients with elevated serum CRP level.

Exp Dermatol 2019 03 11;28(3):240-246. Epub 2019 Feb 11.

Department of Dermatology, Xiangya Hospital, Central South University, Changsha, China.

Chronic spontaneous urticaria (CSU) is a frequent disorder with recurrent itchy wheals and/or angioedema, and nearly 35% patients respond poorly to non-sedating H1 antihistamine treatment. CRP gene encodes the C-reactive protein, which is involved in the pathogenesis of CSU. To investigate the impacts of CRP polymorphisms on the susceptibility and therapeutic efficacy in the South Han CSU patients, we enrolled 145 CSU patients in our study. After 4-week non-sedating H1 antihistamine monotherapy treatment, more than 50% reduction of the severity score is considered as effective, or else non-effective. The CRP rs3093059T/C and rs2794521G/A genotypes of patients were determined by Sequenom MassARRAY. Functional studies including relative luciferase assay and β-hexosaminidase assay were conducted in HEK293T cells or RBL-2H3 cells to explore the function of variants. Forty (62.50%) CSU patients were effective when treated with mizolastine, and 55 (72.4%) patients were effective in the desloratadine group. We found that the patients carried with rs3093059TT genotype were significantly associated with good response (OR = 4.20, P = 0.015), had lower serum CRP, IL-6 and TNF-α levels than the CT/CC genotypes. In vitro, the rs3093059C allele exhibited significantly higher luciferase activity than wild allele (P < 0.001). From the β-hexosaminidase assay, we observed the inhibiting degranulation effects by mizolastine and this effect is weakened when with a higher dose CRP in RBL-2H3 cells. Our findings suggested that CSU patients carrying the rs3093059C allele may respond poorly to mizolastine with elevated serum CRP level.
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http://dx.doi.org/10.1111/exd.13874DOI Listing
March 2019

Discovery and Qualification of Candidate Urinary Biomarkers of Disease Activity in Lupus Nephritis.

J Proteome Res 2019 03 3;18(3):1264-1277. Epub 2019 Jan 3.

Lupus nephritis (LN) is a severe clinical manifestation of systemic lupus erythematosus (SLE) associated with significant morbidity and mortality. Assessment of severity and activity of renal involvement in SLE requires a kidney biopsy, an invasive procedure with limited prognostic value. Noninvasive biomarkers are needed to inform treatment decisions and to monitor disease activity. Proteinuria is associated with disease progression in LN; however, the composition of the LN urinary proteome remains incompletely characterized. To address this, we profiled LN urine samples using complementary mass spectrometry-based methods:  protein gel fractionation, chemical labeling using tandem mass tags, and data-independent acquisition. Combining results from these approaches yielded quantitative information on 2573 unique proteins in urine from LN patients. A multiple-reaction monitoring (MRM) method was established to confirm eight proteins in an independent cohort of LN patients, and seven proteins (transferrin, α-2-macroglobulin, haptoglobin, afamin, α-1-antitrypsin, vimentin, and ceruloplasmin) were confirmed to be elevated in LN urine compared to healthy controls. In this study, we demonstrate that deep mass spectrometry profiling of a small number of patient samples can identify high-quality biomarkers that replicate in an independent LN disease cohort. These biomarkers are being used to inform clinical biomarker strategies to support longitudinal and interventional studies focused on evaluating disease progression and treatment efficacy of novel LN therapeutics.
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http://dx.doi.org/10.1021/acs.jproteome.8b00874DOI Listing
March 2019

MicroRNA‑181 serves an oncogenic role in breast cancer via the inhibition of SPRY4.

Mol Med Rep 2018 Dec 22;18(6):5603-5613. Epub 2018 Oct 22.

Department of Oncology, Xiangya Hospital of Central South University, Changsha, Hunan 410008, P.R. China.

Numerous microRNAs (miRs) have been implicated in breast cancer; however, the molecular mechanism is not fully understood. The present study examined the function and regulatory mechanism of miR‑181 in breast cancer. Reverse transcription‑quantitative polymerase chain reaction and western blot analysis were used to examine the RNA and protein expression. MTT assay, wound healing assay and transwell assay were conducted to study cell proliferation, migration and invasion. Luciferase reporter gene assay was used to confirm targeting relationship. The results suggested that the miR‑181 expression levels were significantly higher in breast cancer cell lines and clinical tissue samples. The increased expression of miR‑181 was markedly associated with higher clinical stage and lymph node metastasis. The patients with high miR‑181 expression demonstrated worse prognosis compared with those with a low expression of miR‑181. Small interfering RNA‑induced miR‑181 downregulation significantly inhibited breast cancer cell proliferation, migration and invasion in vitro, and tumor growth in vivo. Protein sprouty homolog 4 (SPRY4), downregulated in breast cancer tissues and cell lines, was observed to be a novel target gene of miR‑181. Downregulation of SPRY4 was significantly associated with breast cancer progression in addition to poor prognosis. Knockdown of SPRY4 rescued the inhibitory effects of miR‑181 downregulation on the malignant phenotypes of breast cancer cells. Thus, the present study demonstrated that miR‑181 serves a promoting role in breast cancer at least in part through the inhibition of SPRY4 expression. The present results expand the understanding of the miR‑181/SPRY4 axis' function during for the malignant progression of breast cancer.
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http://dx.doi.org/10.3892/mmr.2018.9572DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6236310PMC
December 2018

['s academic thoughts on moxibustion].

Zhongguo Zhen Jiu 2018 Aug;38(8):895-8

Graduate School of Anhui University of CM, Hefei 230038,China.

Xin'an doctor discussed acupuncture and moxibustion on ' and , calling it "orthodox needling method". 's academic thoughts on moxibustion were mainly reflected in () and (). made an incisive exposition of moxibustion treatment of heat syndrome, ulcer and other issues, and there is a corresponding test case for mutual reference. He believed that the moxibustion method only adapts to sick patients and holds opposition to moth moxibustion care for those who were disease-free; he also advocated that detailed verification of different syndrome should be conducted before acupuncture. This paper expounds 's academic thoughts on moxibustion can be used for heat syndrome, moxibustion does not stay phlegm, moxibustion need syndrome and moxibustion treatment of ulcer. It is believed that can present his own unique insights on the basis of his successors. He had played a guiding and promoting role in the development of academic theory of moxibustion.
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http://dx.doi.org/10.13703/j.0255-2930.2018.08.026DOI Listing
August 2018

A Comparison of the Immunostimulatory Effects of Polysaccharides from Tetraploid and Diploid .

Biomed Res Int 2018 9;2018:8628531. Epub 2018 Jul 9.

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangzhou 510642, China.

Polyploidization is an effective means of improving the active components and quality of secondary metabolism in medicinal plants. In the present study, we compared the immunostimulatory effects of crude polysaccharides from tetraploid and diploid . The results showed that the carbohydrate contents of crude polysaccharide of tetraploid (CPE4) and diploid (CPE2) were 85.51% and 44.65%, respectively. H-nuclear magnetic resonance (NMR) spectroscopy and gel-permeation chromatography (GPC) analyses showed no major differences in the overall structure and molecular weight of polysaccharides between CPE4 and CPE2. However, some differences in the relative content of the same polysaccharides group were observed between CPE4 and CPE2. In in vitro tests, EP4 could stimulate lymphocyte proliferation and secretion of cytokines maximally at the concentration of 0.0312 mg/mL, and EP2 could stimulate lymphocyte proliferation and secretion of cytokines maximally at the concentration of 0.125 mg/mL. In in vivo tests, EP4 was more effective at promoting the proliferation of lymphocytes and secretion of cytokines in mice immunosuppressed by cyclophosphamide than EP2 at the same concentration. Taken together, these data demonstrated that the relative content of the partial polysaccharides group is increased, and the immunoregulatory effect is enhanced in tetraploid .
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http://dx.doi.org/10.1155/2018/8628531DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6076949PMC
January 2019

The functional pathway analysis and clinical significance of miR-20a and its related lncRNAs in breast cancer.

Cell Signal 2018 11 6;51:152-165. Epub 2018 Aug 6.

Department of Radiology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China. Electronic address:

Background: miR-20a is a critical molecule in various biological processes and cancer progression procedures. However, its relationships with lncRNAs and their functional pathway analysis in breast tumorigenesis are less intensively studied.

Methods: The expression data from TCGA database and multiple bioinformatics resources were used to check the expression levels, survival curves, interactions and functional illustrations of miR-20a and its related lncRNAs (XIST, H19 and MALAT1) in breast cancer patients. The luciferase reporter assays and Pearson's correlation analyses were utilized to verify the direct regulatory relationship between miR-20a and three lncRNAs (XIST, H19 and MALAT1). In vitro cell proliferation, migration and invasion assays, were performed to check the biological effects of miR-20a and XIST in different breast cancer cell lines. The receiver operating characteristic curve (ROC) analyses were done for evaluating diagnostic values of serum miR-20a and XIST in breast cancer patients.

Results: The miR-20a expression was significantly up-regulated in both breast cancer samples and serum samples, and correlated with poor survival rate in breast cancer patients. LncRNAs (XIST, H19 and MALAT1) directly bound to hsa-miR-20a and were negatively correlated with hsa-miR-20a expression in breast cancer patient samples. For functional illustrations and downstream signaling pathways analysis, XIST, H19 and MALAT1 mainly shared their regulatory functions in cell motility and interleukin signaling in breast cancer progression. Additionally, over-expression of miR-20a and inhibition of XIST promoted breast cancer cell growth, migration and invasion in vitro, and serum miR-20a and XIST served as potential diagnostic biomarkers for breast cancer with the area under ROC curve (AUC) of 0.87 (95% CI = 0.78 to 0.97), and 0.78 (95% CI = 0.67 to 0.89) respectively.

Conclusions: Taken together, these findings provide us novel insights and avenues for utilizing miR-20a and its related lncRNAs as potential diagnostic biomarkers and promising therapeutic targets for breast cancer treatment.
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http://dx.doi.org/10.1016/j.cellsig.2018.08.004DOI Listing
November 2018

In situ fluorescence monitoring of diagnosis and treatment: a versatile nanoprobe combining tumor targeting based on MUC1 and controllable DOX release by telomerase.

Chem Commun (Camb) 2018 Jul;54(59):8277-8280

College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Institute of Molecular and Nano Science, Shandong Normal University, Jinan, 250014 Shandong, P. R. China.

We have constructed versatile drug-loaded nanoprobes capable of responding to both MUC1 and telomerase and achieving intracellular drug release. Besides, the synthesized drug-loaded nanoprobes can realize the in situ imaging observation of the whole process of nanoprobes targeting the tumor cell membrane, the transmembrane entering the cytoplasm and the release of DOX into the cell nucleus.
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http://dx.doi.org/10.1039/c8cc05052jDOI Listing
July 2018