Publications by authors named "Ying Hu"

1,250 Publications

  • Page 1 of 1

Clinical practice guidelines for endoscopic breast surgery in patients with early-stage breast cancer: Chinese Society of Breast Surgery (CSBrS) practice guidelines 2021.

Chin Med J (Engl) 2021 Oct 19. Epub 2021 Oct 19.

Department of Breast Surgery, First Affiliated Hospital of Army Military Medical University, Chongqing 400038, China Department of General Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China.

View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1097/CM9.0000000000001592DOI Listing
October 2021

A deep learning-and CT image-based prognostic model for the prediction of survival in non-small cell lung cancer.

Med Phys 2021 Oct 18. Epub 2021 Oct 18.

School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, 200000, China.

Objective: To assist clinicians in arranging personalized treatment, planning follow-up programs and extending survival times for non-small cell lung cancer (NSCLC) patients, a method of deep learning combined with computed tomography (CT) imaging for survival prediction was designed.

Methods: Data were collected from 484 patients from 4 research centers. The data from 344 patients were utilized to build the A_CNN survival prognosis model to classify 2-year overall survival time ranges (730 days cut off). Data from 140 patients, including independent internal and external test sets, were utilized for model testing. First, a series of preprocessing techniques were used to process the original CT images and generate training and test data sets from the axial, coronal, and sagittal planes. Second, the structure of the A_CNN model was designed based on asymmetric convolution, bottleneck blocks, the uniform cross-entropy (UC) loss function, and other advanced techniques. After that, the A_CNN model was trained, and numerous comparative experiments were designed to obtain the best prognostic survival model. Last, the model performance was evaluated, and the predicted survival curves were analyzed.

Results: The A_CNN survival prognosis model yielded a high patient-level accuracy of 88.8%, a patch-level accuracy of 82.9%, and an area under the receiver operating characteristic (ROC) curve (AUC) of 0.932. When tested on an external data set, the maximum patient-level accuracy was 80.0%.

Conclusions: The results suggest that using a deep learning method can improve prognosis in patients with NSCLC and has important application value in establishing individualized prognostic models. This article is protected by copyright. All rights reserved.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/mp.15302DOI Listing
October 2021

Catalpol alleviates Ang II-induced renal injury through NF/κB pathway and TGF-β1/Smads pathway.

J Cardiovasc Pharmacol 2021 Oct 4. Epub 2021 Oct 4.

Department of Cardiology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, P.R. China Department of Traumatic Orthopaedics, The Third Affiliated Hospital of Shandong First Medical University (Affiliated Hospital of Shandong Academy of Medical Sciences), Jinan, Shandong, 250031, P.R. China Department of science and technology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, P.R. China Department of Nephrology, The Third Affiliated Hospital of Shandong First Medical University (Affiliated Hospital of Shandong Academy of Medical Sciences), Jinan, Shandong, 250031, P.R. China.

Abstract: Catalpol is an iridoid glycoside obtained from Rehmannia glutinosa, which in previous studies showed various pharmacological properties, including anti-inflammatory, antioxidant, antidiabetic, antitumor and dopaminergic neurons protecting effects. Here, we examined the effect of catalpol on AngII-induced renal injury induced by angiotensin II (AngII), and further to explore its latent molecular mechanisms. We used an in vivo model of AngII-induced renal injury mice, catalpol (25, 50, and 100 mg/kg) was administered for 28 days. Mouse glomerular mesangial cells (SV40 MES 13), rat kidney interstitial fibroblasts cells (NRK-49F), and human proximal tubular epithelial cells (HK-2) were induced by AngII (10 µM) in the presence or absence of catalpol (1, 5, and 10 µM) and incubated for 48 h in vitro. In our study, PAS and masson staining of renal tissue showed that catalpol reduced AngII-induced renal injury in a concentration-dependent manner. The positive expressions of Collagen IV and TGF-β1 were observed to decrease sharply after catalpol treatment. In renal tissue, the levels of pro-inflammatory cytokines TNF-α and IL-6 were evidently decreased after catalpol intervention. Catalpol can relieve AngII-induced renal injury by inactivating NF/κB and TGF-β1/Smads signaling pathways. Therefore, catalpol may act as a potential drug to treat AngII-induced renal injury.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1097/FJC.0000000000001148DOI Listing
October 2021

In vitro anti-motile effects of Rhoifolin, a flavonoid extracted from Callicarpa nudiflora on breast cancer cells via downregulating Podocalyxin-Ezrin interaction during Epithelial Mesenchymal Transition.

Phytomedicine 2021 Jan 28;93:153486. Epub 2021 Jan 28.

School of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. Electronic address:

Background: Callicarpa nudiflora (C. nudiflora), which is a medical herb in genus of Callicarpa, widely grows in the southern part of China. Several investigations had shown that this herb exerts anti-tumor effects. Ezrin is an important membrane-cytoskeleton-binding protein. By organizing membrane proteins and orchestrating their signal transduction, Ezrin contributes to modulation of cytoskeleton rearrangement in cell motility.

Purpose: To investigate the anti-motile properties of Rhoifolin (RFL), a flavonoid from C. nudiflora, and to determine whether its effects are related to the inhibition on Podocalyxin (PODXL)-Ezrin signal transduction.

Methods: To determine suitable concentration of RFL and exposure time on breast cancer cells, the effects of RFL on viability of breast cancer cells were evaluated by MTT assay. Then, the anti-migratory properties of RFL were determined by AP 48 chamber system and ORIS cell migration assay. F-actin in MDA-MB-231 cells was visualized by Alexa Fluor™ 488 conjugated Phalloidin. Immunoprecipitation was involved to access the effects of RFL on the interaction between Ezrin and PODXL. In addition, several EMT markers, including E-cadherin, Vimentin, Snail and Slug, were measured by Western Blotting assay and cell immunofluorescent analysis. Finally, the effects of RFL on cell migration, expression of Ezrin and EMT markers were verified by small interfering RNA (siRNA) mediated gene silencing.

Results: We showed here that treatments with 10 and 40 μM of RFL induced significant inhibitions on cell migration and alterations on the location and organization of actin cytoskeleton in breast cancer cells. Next, it was found that RFL suppressed Ezrin phosphorylation and consequent interaction with PODXL, significantly. Also, this compound showed an obvious inhibitory effect on TGF-β1-induced EMT in MDA-MB-231 cells. Furthermore, data from RNA interfering assay confirmed that the inhibitory effects of RFL on Ezrin was enhanced by the deletion of Ezrin.

Conclusion: RFL shows anti-motile properties on breast cancer cells, which is due to its potential to downregulate Podocalyxin-Ezrin interaction during Epithelial Mesenchymal Transition.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.phymed.2021.153486DOI Listing
January 2021

De-Doped Polyaniline as a Mediating Layer Promoting In-Situ Growth of Metal-Organic Frameworks on Cellulose Fiber and Enhancing Adsorptive-Photocatalytic Removal of Ciprofloxacin.

Polymers (Basel) 2021 Sep 27;13(19). Epub 2021 Sep 27.

Key Laboratory of Bio-Based Material Science & Technology, Northeast Forestry University, Ministry of Education, Harbin 150040, China.

New kinds of inorganic-organic hybrid porous materials, metal-organic frameworks (MOFs), have shown great application potential in various fields, but their powdery nature limits their application to a certain extent. As a green and renewable biomass material in nature, cellulose fiber (CelF) has the advantages of biodegradability, recyclability and easy processing, and can be used as an excellent flexible substrate for MOFs. However, the efficient deposition of MOFs on CelF is still a great challenge for the development of this new material. Herein, polyaniline (PANI) and de-doped PANI (DPANI) with rich functional groups as a mediating layer was proposed to promote the in-situ growth and immobilization of some MOFs on CelF. The PANI (especially DPANI) layer greatly promoted the deposition of the four MOFs, and more encouragingly, significantly promoted the in-situ growth and nanocrystallization of MIL-100(Fe). MIL-100(Fe)@[email protected] was selected as an adsorbent-photocatalyst to be used for the adsorptive-photocatalytic removal of ciprofloxacin (CIP) in water. The removal efficiency of CIP by MIL-100(Fe)@[email protected] reached 82.78%, and the removal capacity of CIP was as high as 105.96 mg g. The study found that DPANI had a synergistic effect on both the in-situ growth of MIL-100(Fe) on CelF and the adsorption-photocatalysis of CIP in water. The universal platform of PANI-mediated in-situ growth and immobilization of MOFs on CelF constructed in this study widens the road for the development of [email protected] composites.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/polym13193298DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8512102PMC
September 2021

Carbonyl flavor compound-targeted colorimetric sensor array based on silver nitrate and o-phenylenediamine derivatives for the discrimination of Chinese Baijiu.

Food Chem 2021 Sep 22;372:131216. Epub 2021 Sep 22.

College of Chemical Engineering, Zhejiang University of Technology, Hangzhou 310032, PR China. Electronic address:

Baijiu is a distilled liquor of great importance in the food industry. Various aroma types, brands, and grades of Baijiu have filled the market; thus, discrimination for quality control is required. Herein, we constructed a novel colorimetric sensor array based on the redox reaction between silver nitrate and o-phenylenediamine or its derivatives for the discrimination of carbonyl flavor compounds (CFCs) and Baijius. The specific colored products were changed by CFCs depending on the influence of silver nanoparticle aggregation and chemical reactions. The array was used to qualitatively and quantitatively identify 21 CFCs with fast response (<14 min), wide linear range (0.025-25 mmol/L), and low detection limits (<60 μmol/L, 29 nmol/L for carboxylic acids). Finally, the array was successfully applied to the discrimination of 56 Baijius. The method proposed in this study is simple, fast, reliable, and has good application potential for the visual determination of Chinese Baijiu.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.foodchem.2021.131216DOI Listing
September 2021

The MSDB sends a GABAergic projection to cholinergic neurons in the ventral MHb.

Mol Psychiatry 2021 Jul;26(7):2679

Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI, USA.

View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41380-021-01280-1DOI Listing
July 2021

Systematic review and meta-analysis of the correlation between intestinal flora and gestational diabetes mellitus.

Ann Palliat Med 2021 Sep;10(9):9752-9764

Department of Gynecology and Obstetrics, Zhejiang Hospital, Hangzhou, China.

Background: A meta-analysis was conducted on the correlation between intestinal flora and gestational diabetes mellitus (GDM) to provide a theoretical basis for the treatment of GDM.

Methods: The PubMed, Embase, MEDLINE, Ovid, Springer, and Web of Science databases were searched from the establishment of the databases to December 31, 2020, to retrieve randomized control trials (RCTs) involving GDM patients. The Cochrane Handbook for Systematic Reviews of Intervention 5.0.2 was used to assess the bias risk of the included articles, and Rev Man 5.3 was used for the meta-analysis.

Results: A total of 7 studies were included in the meta-analysis, comprising 665 study participants. The meta-analysis results showed that the GDM patients in the experimental group had a lower level of Bifidobacterium [MD (mean difference) =-2.49; 95% confidence interval (CI): -3.54 to -1.45; Z=4.66; P<0.00001], Lactobacillus [MD =-1.69; 95% CI: -1.84 to -1.53; Z=20.66; P<0.00001], Bacteroides [MD =-1.17; 95% CI: -1.45 to -0.89; Z=8.15; P<0.00001], Bacteroidetes [MD =-1.22; 95% CI: -1.71 to -0.72; Z=4.81; P<0.00001], and a higher level of Enterobacter [MD =1.79; 95% CI: 1.13 to 2.45; Z=5.3; P<0.00001], Enterococcus [MD =1, 29; 95% CI: 0.98 to 1.6; Z=8.06; P<0.00001], Fusobacterium [MD =0.03; 95% CI: -0.13 to 0.19; Z=0.37; P=0.71], tumor necrosis factor alpha (TNF-α) [MD =113.66; 95% CI: 52.01 to 175.31; Z=3.61; P=0.0003], interleukin (IL)-17 [MD =37.92; 95% CI: 29.74 to 46.1; Z=9.09; P<0.00001], and IL-6 [MD =66.38; 95% CI: 33.6 to 99.15; Z=3.97; P<0.0001] than those in the control group; however, no statistically significant difference was found in relation to Fusobacterium between the experimental group and the control group.

Discussion: Intestinal microecological changes are closely related to the occurrence of GDM in our study, which manifested as a decrease in the level of probiotics, an increase in the level of intestinal bacteria and other strains, and an increase in the level of inflammatory factors. Thus, special attention should be paid to changes in patients' intestinal flora to prevent GDM.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.21037/apm-21-2061DOI Listing
September 2021

Protein-protein interaction relation extraction based on multigranularity semantic fusion.

J Biomed Inform 2021 Oct 8;123:103931. Epub 2021 Oct 8.

Department of Computer Science and Technology, Xi'an Jiaotong University, Xi'an, China.

Extracting semantic relationships about biomedical entities in a sentence is a typical task in biomedical information extraction. Because a sentence usually contains several named entities, it is important to learn global semantics of a sentence to support relation extraction. In related works, many strategies have been proposed to encode a sentence representation relevant to considered named entities. Despite the current success, according to the characteristic of languages, semantics of words are expressed on multigranular levels which also heavily depends on local semantic of a sentence. In this paper, we propose a multigranularity semantic fusion method to support biomedical relation extraction. In this method, Transformer is adopted for embedding words of a sentence into distributed representations, which is effective to encode global semantic of a sentence. Meanwhile, a multichannel strategy is applied to encode local semantics of words, which enables the same word to have different representations in a sentence. Both global and local semantic representations are fused to enhance the discriminability of the neural network. To evaluate our method, experiments are conducted on five standard PPI corpora (AImed, BioInfer, IEPA, HPRD50, and LLL), which achieve F1-scores of 83.4%, 89.9%, 81.2%, 84.5%, and 92.5%, respectively. The results show that multigranular semantic fusion is helpful to support the protein-protein interaction relationship extraction.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jbi.2021.103931DOI Listing
October 2021

Bipolar disorder cured by vitamin D supplementation in a 15-year-old boy: A case report.

Bipolar Disord 2021 Oct 8. Epub 2021 Oct 8.

Clinical Pharmacy Center (Pharmacy Department), Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Zhejiang, China.

View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/bdi.13143DOI Listing
October 2021

mmBCFA C17iso ensures endoplasmic reticulum integrity for lipid droplet growth.

J Cell Biol 2021 Nov 8;220(11). Epub 2021 Oct 8.

Center for Life Sciences, School of Life Sciences, Yunnan University, Kunming, Yunnan, China.

In eukaryote cells, lipid droplets (LDs) are key intracellular organelles that dynamically regulate cellular energy homeostasis. LDs originate from the ER and continuously contact the ER during their growth. How the ER affects LD growth is largely unknown. Here, we show that RNAi knockdown of acs-1, encoding an acyl-CoA synthetase required for the biosynthesis of monomethyl branched-chain fatty acids C15iso and C17iso, remarkably prevented LD growth in Caenorhabditis elegans. Dietary C17iso, or complex lipids with C17iso including phosphatidylcholine, phosphatidylethanolamine, and triacylglycerol, could fully restore the LD growth in the acs-1RNAi worms. Mechanistically, C17iso may incorporate into phospholipids to ensure the membrane integrity of the ER so as to maintain the function of ER-resident enzymes such as SCD/stearoyl-CoA desaturase and DGAT2/diacylglycerol acyltransferase for appropriate lipid synthesis and LD growth. Collectively, our work uncovers a unique fatty acid, C17iso, as the side chain of phospholipids for determining the ER homeostasis for LD growth in an intact organism, C. elegans.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1083/jcb.202102122DOI Listing
November 2021

Characterization of the complete chloroplast genome of (Caprifoliaceae).

Mitochondrial DNA B Resour 2021 27;6(10):3067-3069. Epub 2021 Sep 27.

Guangxi Key Laboratory of Medicinal Resources Protection and Genetic Improvement, Guangxi Botanical Garden of Medicinal Plants, Nanning, China.

The complete chloroplast genome of Hemsl. has been characterized by reference-based assembly using Illumina paired-end data. The circular complete cp genome is 155,463 bp in length, containing a large single copy (LSC) region of 89,282 bp, a small single copy (SSC) region of 18,661 bp, which are separated by a pair of inverted repeat (IR) regions of 23,760 bp. A total of 129 genes were predicted from the cp genome, including 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Phylogenetic analysis reveals that is more closely related to Thunb. and Rehd. Our result will provide a reference for the phylogenetic relationship, plant identification and resource development and utilization of species.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/23802359.2021.1981161DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8480604PMC
September 2021

Case Report: Efficiency of Embolization Microcoils for the Repair of Brachiocephalic Vein Perforation During Hemodialysis Catheter Placement.

Front Med (Lausanne) 2021 17;8:726120. Epub 2021 Sep 17.

Department of Nephrology, Kidney Research Institute, West China Hospital of Sichuan University, Chengdu, China.

The cuff catheter is one of the most common routes of vascular access in hemodialysis patients, while severe complications can occur during cuff catheter placement, such as bleeding, hematoma, and artery or vein damage. During catheterization, brachiocephalic vein perforation associated with a mediastinal lesion is rare. Open chest repair is effective for brachiocephalic vein perforation during catheter placement, but it entails a risk of potentially lethal trauma. Interventional treatment can be considered to reduce injury in this context, but relevant reports are limited. Herein, we describe our experience with a 68-year-old male hemodialyzed patient in whom cuff catheter vascular access was required for regular hemodialysis. He complained of mild pain in the left side of his chest during cuff catheter placement. The surgeon immediately checked the location of the catheter. Digital subtraction angiography revealed that the hemodialysis cuff catheter had punctured the mediastinal area from the left brachiocephalic vein. The patient was diagnosed with left brachiocephalic vein perforation (d ≈ 5 mm). Fortunately, the brachiocephalic vein perforation was successfully repaired with two embolization microcoils after comprehensive assessment and multidisciplinary consultation. Brachiocephalic vein perforation can be repaired with embolization microcoils during hemodialysis catheter placement, and this method of interventional treatment is safe and effective.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fmed.2021.726120DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8484305PMC
September 2021

Case Report: Efficiency of Embolization Microcoils for the Repair of Brachiocephalic Vein Perforation During Hemodialysis Catheter Placement.

Front Med (Lausanne) 2021 17;8:726120. Epub 2021 Sep 17.

Department of Nephrology, Kidney Research Institute, West China Hospital of Sichuan University, Chengdu, China.

The cuff catheter is one of the most common routes of vascular access in hemodialysis patients, while severe complications can occur during cuff catheter placement, such as bleeding, hematoma, and artery or vein damage. During catheterization, brachiocephalic vein perforation associated with a mediastinal lesion is rare. Open chest repair is effective for brachiocephalic vein perforation during catheter placement, but it entails a risk of potentially lethal trauma. Interventional treatment can be considered to reduce injury in this context, but relevant reports are limited. Herein, we describe our experience with a 68-year-old male hemodialyzed patient in whom cuff catheter vascular access was required for regular hemodialysis. He complained of mild pain in the left side of his chest during cuff catheter placement. The surgeon immediately checked the location of the catheter. Digital subtraction angiography revealed that the hemodialysis cuff catheter had punctured the mediastinal area from the left brachiocephalic vein. The patient was diagnosed with left brachiocephalic vein perforation (d ≈ 5 mm). Fortunately, the brachiocephalic vein perforation was successfully repaired with two embolization microcoils after comprehensive assessment and multidisciplinary consultation. Brachiocephalic vein perforation can be repaired with embolization microcoils during hemodialysis catheter placement, and this method of interventional treatment is safe and effective.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fmed.2021.726120DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8484305PMC
September 2021

MLIP causes recessive myopathy with rhabdomyolysis, myalgia and baseline elevated  serum creatine kinase.

Brain 2021 Oct;144(9):2722-2731

Department of Chemistry and Earth Science, College of Arts and Sciences, Qatar University, Qatar.

Striated muscle needs to maintain cellular homeostasis in adaptation to increases in physiological and metabolic demands. Failure to do so can result in rhabdomyolysis. The identification of novel genetic conditions associated with rhabdomyolysis helps to shed light on hitherto unrecognized homeostatic mechanisms. Here we report seven individuals in six families from different ethnic backgrounds with biallelic variants in MLIP, which encodes the muscular lamin A/C-interacting protein, MLIP. Patients presented with a consistent phenotype characterized by mild muscle weakness, exercise-induced muscle pain, variable susceptibility to episodes of rhabdomyolysis, and persistent basal elevated serum creatine kinase levels. The biallelic truncating variants were predicted to result in disruption of the nuclear localizing signal of MLIP. Additionally, reduced overall RNA expression levels of the predominant MLIP isoform were observed in patients' skeletal muscle. Collectively, our data increase the understanding of the genetic landscape of rhabdomyolysis to now include MLIP as a novel disease gene in humans and solidifies MLIP's role in normal and diseased skeletal muscle homeostasis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/brain/awab275DOI Listing
October 2021

TRPM7 is critical for short-term synaptic depression by regulating synaptic vesicle endocytosis.

Elife 2021 09 27;10. Epub 2021 Sep 27.

Department of Biological Sciences, University of Illinois at Chicago, Chicago, United States.

Transient receptor potential melastatin 7 (TRPM7) contributes to a variety of physiological and pathological processes in many tissues and cells. With a widespread distribution in the nervous system, TRPM7 is involved in animal behaviors and neuronal death induced by ischemia. However, the physiological role of TRPM7 in central nervous system (CNS) neuron remains unclear. Here, we identify endocytic defects in neuroendocrine cells and neurons from TRPM7 knockout (KO) mice, indicating a role of TRPM7 in synaptic vesicle endocytosis. Our experiments further pinpoint the importance of TRPM7 as an ion channel in synaptic vesicle endocytosis. Ca imaging detects a defect in presynaptic Ca dynamics in TRPM7 KO neuron, suggesting an importance of Ca influx via TRPM7 in synaptic vesicle endocytosis. Moreover, the short-term depression is enhanced in both excitatory and inhibitory synaptic transmissions from TRPM7 KO mice. Taken together, our data suggests that Ca influx via TRPM7 may be critical for short-term plasticity of synaptic strength by regulating synaptic vesicle endocytosis in neurons.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.7554/eLife.66709DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8516418PMC
September 2021

Curcumin inhibits proliferation and invasion of papillary thyroid carcinoma cells by inhibiting the JAK2 / STAT3 pathway.

J BUON 2021 Jul-Aug;26(4):1635-1641

Clinical Pharmacy Center, Department of Pharmacy, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, China.

Purpose: The purpose of this study was to analyze the function of curcumin to suppress the proliferative and invasive abilities of papillary thyroid carcinoma (PTC) through inhibiting the JAK2/STAT3 pathway.

Methods: After treatment of different doses of curcumin in TPC-1 and SW1736 cells, changes in viability, clonality, cell cycle, apoptosis, wound healing and invasion were determined. Western blot analyses were performed to detect protein levels of apoptosis-associated genes, JAK2 and STAT3 in TPC-1 and SW1736 cells treated with different doses of curcumin.

Results: Curcumin treatment dose-dependently reduced viability, clonality and metastatic ability in TPC-1 and SW1736 cells. After treatment of 10 μM or 20 μM curcumin, PTC cells were blocked in G2/M phase, and their apoptotic rate increased. Curcumin treatment downregulated Bcl-2 and upregulated Bax in PTC cells. In addition, curcumin treatment downregulated p-JAK2 and p-STAT3 in TPC-1 and SW1736 cells.

Conclusions: Curcumin treatment blocks PTC cells to proliferate and invade via inhibiting the JAK2/STAT3 pathway.
View Article and Find Full Text PDF

Download full-text PDF

Source
September 2021

Recent developments in catalytic cross-couplings with unsaturated carboxylates.

Org Biomol Chem 2021 Sep 22;19(36):7754-7767. Epub 2021 Sep 22.

Key Laboratory of Organic Synthesis of Jiangsu Province, College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Ren-Ai Road 199, 215123 Suzhou, China.

Catalytic cross-couplings through C-O bond-cleavage of unsaturated carboxylates with organometallics have emerged as a powerful method for sustainable syntheses. Over the last decade, remarkable achievements have been made with the development of transition metal-catalyzed cross-couplings with the readily available phenol and enol derivatives as suitable coupling electrophiles beyond unsaturated halides. Therefore, this perspective describes the recent advances in the field of transition metal-catalyzed C-O bond activation of unsaturated carboxylates with organometallics, including B, Mg, Zn, Al, and Si reagents, until May 2021.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1039/d1ob00955aDOI Listing
September 2021

Busulfan impairs blood-testis barrier and spermatogenesis by increasing noncollagenous 1 domain peptide via matrix metalloproteinase 9.

Andrology 2021 Sep 18. Epub 2021 Sep 18.

Department of Biochemistry and Molecular Biology, College of Life Science, China Medical University, Shenyang, China.

Backgrounds: Sterility induced by anti-cancer treatments has caused significant concern, yet the mechanism and treatment exploration are little for male infertility after cancer therapy. Busulfan, the antineoplastic that was widely applied before bone marrow transplantation, was known to induce male reproductive disorder.

Objectives: To investigate the effect of busulfan on blood-testis barrier function in adult rats and determine whether noncollagenous 1 domain peptide, the biologically active fragment proteolyzed from the collagen α3 chain (IV) by matrix metalloproteinase 9, was involved during this process.

Materials And Methods: Adult male rats were treated with one-dose or double-dose of busulfan (10 mg/kg) before euthanized at day 35. Blood-testis barrier integrity assay, HE staining, immunofluorescence, and Western blot were used to validate the effect of busulfan on blood-testis barrier permeability and spermatogenesis. JNJ0966 was applied to specifically inhibit the matrix metalloproteinase 9 activity. The polymerization activity of F-actin/G-actin and microtubule/tubulin in the testis were assessed by using commercial kits.

Results: A noteworthy blood-testis barrier injury and significant up-regulation of matrix metalloproteinase 9 activity and noncollagenous 1 level after a single-dose busulfan (10 mg/kg) treatment in adult rat testis were revealed. The application of JNJ0966 was found to decrease noncollagenous 1 level and rescue the busulfan-induced blood-testis barrier injury including the mis-localization of junction proteins across the seminiferous epithelium, by recovering the organization and polymerization of both F-actin and microtubule. The busulfan-induced spermatogenesis impairment was also improved by JNJ0966.

Conclusion: These findings thus demonstrate that the elevation in matrix metalloproteinase 9 and noncollagenous 1 might participate in busulfan-induced blood-testis barrier disruption in adult male rats. As such, busulfan-induced male infertility could possibly be managed through interventions on noncollagenous 1 production.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/andr.13112DOI Listing
September 2021

Bioflavonoid Suppresses Hypertrophic Scar Formation by the TGF-/Smad Signaling Pathway.

Evid Based Complement Alternat Med 2021 4;2021:2444839. Epub 2021 Sep 4.

Department of Traditional Chinese Medicine, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510630, China.

Background: Hypertrophic scar (HS) is a benign fibroproliferative skin disease resulting from an aberrant wound healing process and can cause aesthetic and functional damage to patients. Currently, there is no ideal treatment to treat this disease. , a natural active bioflavonoid compound, is suggested to inhibit fibrosis and proliferation in certain cells.

Methods: In this study, we found could attenuate abnormal scar formation in an HS rabbit ear model. Additionally, the HE staining shows reduced scar elevation index (SEI) and Masson's trichrome staining changed collagen deposition.

Results: The expressions of type I collagen, type III collagen, and TGF-1 were much lower under a proper dose of treatment, and Smad7 expression was also enhanced, which are examined by real-time PCR, immunohistochemistry, and western blot.

Conclusion: Our data indicated that can alleviate dermal scarring via the TGF-/Smad signaling pathway probably by upregulating Smad 7 expression and, thus, suppressing the expression of type I and type III collagens and TGF-1.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1155/2021/2444839DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8434902PMC
September 2021

Case Report: CNNM2 Mutations Cause Damaged Brain Development and Intractable Epilepsy in a Patient Without Hypomagnesemia.

Front Genet 2021 20;12:705734. Epub 2021 Aug 20.

Department of Pediatric Neurology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.

A series of neurological manifestations such as intellectual disability and epilepsy are closely related to hypomagnesemia. Cyclin M2 (CNNM2) proteins, as a member of magnesium (Mg) transporters, were found along the basolateral membrane of distal renal tubules and involved in the reabsorption of Mg. Homozygous and heterozygous variants in reported so far were responsible for a variable degree of hypomagnesemia, several of which also showed varying degrees of neurological phenotypes such as intellectual disability and epilepsy. Here, we report a heterozygous variant (c.2228C > T, p.Ser743Phe) in a Chinese patient, which is the variant located in the cyclic nucleotide monophosphate-binding homology (CNBH) domain of CNNM2 proteins. The patient presented with mild intellectual disability and refractory epilepsy but without hypomagnesemia. Thus, we reviewed the literature and analyzed the phenotypes related to variants, and then concluded that the number of variant alleles and the changed protein domains correlates with the severity of the disease, and speculated that the CNBH domain of CNNM2 possibly plays a limited role in Mg transport but a significant role in brain development. Furthermore, it can be speculated that neurological phenotypes such as intellectual disability and seizures can be purely caused by CNNM2 variants.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fgene.2021.705734DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8417836PMC
August 2021

Genotyping-By-Sequencing diversity analysis of international Vanilla collections uncovers hidden diversity and enables plant improvement.

Plant Sci 2021 Oct 13;311:111019. Epub 2021 Aug 13.

Horticultural Sciences Department, University of Florida, Gainesville, FL, USA. Electronic address:

Genomics-based diversity analysis of natural vanilla populations is important in order to guide conservation efforts and genetic improvement through plant breeding. Vanilla is a cultivated, undomesticated spice that originated in Mesoamerica prior to spreading globally through vegetative cuttings. Vanilla extract from the commercial species, mainly V. planifolia and V. × tahitensis, is used around the world as an ingredient in foods, beverages, cosmetics, and pharmaceuticals. The global reliance on descendants of a few foundational clones in commercial production has resulted in an industry at heightened risk of catastrophic failure due to extremely narrow genetic diversity. Conversely, national and institutional collections including those near the center of cultivation contain previously undiscovered diversity that could bolster the genetic improvement of vanilla and guide conservation efforts. Towards this goal, an international vanilla genotyping effort generated and analyzed 431,204 single nucleotide polymorphisms among 412 accessions and 27 species from eight collections. Phylogenetic and STRUCTURE analysis sorted vanilla by species and identified hybrid accessions. Principal Component Analysis and the Fixation Index (FST) were used to refine relationships among accessions and showed differentiation among species. Analysis of the commercial species split V. planifolia into three types with all V. × tahitensis accessions being most similar to V. planifolia type 2. Finally, an in-depth analysis of V. × tahitensis identified seven V. planifolia and six V. odorata accessions as most similar to the estimated parental genotypes providing additional data in support of the current hybrid theory. The prevalence of probable V. × tahitensis parental accessions from Belize suggests that V. × tahitensis could have originated from this area and highlights the need for vanilla conservation throughout Central and South America. The genetic groupings among accessions, particularly for V. planifolia, can now be used to focus breeding efforts on fewer accessions that capture the greatest diversity.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.plantsci.2021.111019DOI Listing
October 2021

Huaier polysaccharides suppress triple-negative breast cancer metastasis and epithelial-mesenchymal transition by inducing autophagic degradation of Snail.

Cell Biosci 2021 Sep 4;11(1):170. Epub 2021 Sep 4.

Breast Disease Center, Southwest Hospital, Army Medical University, 30# Gaotanyan street, Chongqing, 400038, China.

Background: Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, and the targeted therapies are lacking for this type of cancer. We previously demonstrated that Huaier effectively improve 5-year OS and DFS in stage III TNBC patients, and the polysaccharides of Huaier (PS-T) have been identified as the major components of Huaier. However, the mechanisms of anti-tumor action of PS-T is unclear. This study aimed to investigate the effect of PS-T on TNBC cell invasion and migration.

Results: This study showed that PS-T inhibited cell invasion and migration both in vitro and in vivo by inducing autophagy to suppress epithelial-mesenchymal transition (EMT). Autophagy inhibitor LY294002 or knockdown of ATG5 suppressed the inhibitory effects of PS-T. In addition, as a key transcription factor controlling EMT initiation, Snail was found to be degraded by PS-T induced autophagy. In addition, overexpression of Snail reversed the inhibitory effects of PS-T. Furthermore, it was confirmed that the expression of Snail was inversely correlated with LC3 and associated with poor prognosis using immunohistochemistry and TCGA database analysis, respectively.

Conclusions: This study demonstrated that PS-T could inhibit EMT in breast cancer cells by inducing autophagy to degrade Snail protein, thus improving the prognosis of TNBC, offering potential treatment alternatives for TNBC patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13578-021-00682-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8417980PMC
September 2021

Levels of anti-oxidative molecules and inflammatory factors in patients with vascular dementia and their clinical significance.

Pak J Med Sci 2021 Sep-Oct;37(5):1509-1513

Juan Gao Department of Neurology, Baoding First Central Hospital, Baoding 071000, Hebei, China.

Objectives: To explore levels of anti-oxidative molecules and inflammatory factors in patients with vascular dementia (VD) and their clinical significance.

Methods: Sixty VD patients admitted in our hospital from January 2016 to January 2019 were classified into an experimental group, while another 60 healthy patients seeking physical examinations in the corresponding period were selected as a control group. Various indexes related to serum inflammatory factors and anti-oxidative molecules were compared among patients in such two groups. For the purpose of comparing anti-oxidative molecular expression levels and inflammatory factor levels in patients with VD of different severities, 60 cases in the experimental group were divided, based on a Mini-mental State Examination (MMSE) scale, into patients with mild symptoms (n=20, score: 21~26), patients with moderate symptoms (n=22, score: 10~20) and patients with severe symptoms (n=18, score: 0~9).

Results: By contrast to the control group, levels of inflammatory factors (e.g., TNF-a, CRP and IL-6) in VD patients are all significantly increased and their differences show statistical significance (p<0.05); and, expression levels of anti-oxidative factors, including superoxide dismutase (SOD), glutathion peroxidase (GSH-Px), total antioxidant capacity (TAC), catalase (CAT) and glutathione reductase (GR), in the experimental group are apparently below those of the control group (P<0.05). As dementia degree increases, expression levels of serum anti-oxidative molecules in such patients are inclined to drop in a significant way (P<0.05), while inflammatory factor levels tend to go up gradually (P<0.05).

Conclusions: If compared with the normal population, inflammatory factor levels in serum of VD patients are higher; however, expression levels of anti-oxidative molecules become below those of the normal population. Additionally, levels of inflammatory factors and anti-oxidative molecules may change obviously as severity of illness increases. This suggests that inflammation and oxidation play a certain role of auxoaction in VD patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.12669/pjms.37.5.3854DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8377923PMC
November 2020

Expression of SGLT1 in the Mouse Endometrial Epithelium and its Role in Early Embryonic Development and Implantation.

Reprod Sci 2021 Nov 30;28(11):3094-3108. Epub 2021 Aug 30.

Department of Physiology, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, No. 17 Section 3 Renmin South Road, Chengdu, 610041, People's Republic of China.

Many functional activities of endometrium epithelium are energy consuming which are very important for maintaining intrauterine environment needed by early embryonic development and establishment of implantation window. Glucose is a main energy supplier and one of the main components of intrauterine fluid. Obviously, glucose transports in endometrium epithelium involve in for these activities but their functions have not been elucidated. In this research, we observed a spatiotemporal pattern of sodium glucose transporter 1 (SGLT1) expression in the mouse endometrium. We also determined that progesterone can promote the expression of SGLT1 in the mouse endometrial epithelium in response to the action of oestrogen. Treatment with the SGLT1 inhibitor phlorizin or small interfering RNA specific for SGLT1 (SGLT1-siRNA) altered glucose uptake in primary cultured endometrial epithelial cells, which exhibited reduced ATP levels and AMPK activation. The injection of phlorizin or SGLT1-siRNA into one uterine horn of each mouse on day 2 of pregnancy led to an increased glucose concentration in the uterine fluid and decreased number of harvested normal blastocysts and decreased expression of integrin αVβ3 in endometrial epithelium and increased expression of mucin 1 and lactoferrin in endometrial epithelium and the uterine homogenates exhibited activated AMPK, a decreased ATP level on day 4, and a decreased number of implantation sites on day 5. In embryo transfer experiments, pre-treatment of the uterine horn with phlorizin or SGLT1-siRNA during the implantation window led to a decreased embryo implantation rate on day 5 of pregnancy, even when embryos from normal donor mice were used. In conclusion, SGLT1, which participates in glucose transport in the mouse endometrial epithelium, inhibition and/or reduced expression of SGLT1 affects early embryo development by altering the glucose concentration in the uterine fluid. Inhibition and/or reduced expression of SGLT1 also affects embryo implantation by influencing energy metabolism in epithelial cells, which consequently influences implantation-related functional activities.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s43032-021-00480-yDOI Listing
November 2021

[Progress in computer-assisted Alberta stroke program early computer tomography score of acute ischemic stroke based on different modal images].

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 2021 Aug;38(4):790-796

Institute of Medical Imaging Engineering, University of Shanghai for Science & Technology, Shanghai 200093, P.R.China.

Clinically, non-contrastive computed tomography (NCCT) is used to quickly diagnose the type and area of ​​stroke, and the Alberta stroke program early computer tomography score (ASPECTS) is used to guide the next treatment. However, in the early stage of acute ischemic stroke (AIS), it's difficult to distinguish the mild cerebral infarction on NCCT with the naked eye, and there is no obvious boundary between brain regions, which makes clinical ASPECTS difficult to conduct. The method based on machine learning and deep learning can help physicians quickly and accurately identify cerebral infarction areas, segment brain areas, and operate ASPECTS quantitative scoring, which is of great significance for improving the inconsistency in clinical ASPECTS. This article describes current challenges in the field of AIS ASPECTS, and then summarizes the application of computer-aided technology in ASPECTS from two aspects including machine learning and deep learning. Finally, this article summarizes and prospects the research direction of AIS-assisted assessment, and proposes that the computer-aided system based on multi-modal images is of great value to improve the comprehensiveness and accuracy of AIS assessment, which has the potential to open up a new research field for AIS-assisted assessment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.7507/1001-5515.202012037DOI Listing
August 2021

mA RNA Methylation Regulator YTHDF1 Correlated With Immune Microenvironment Predicts Clinical Outcomes and Therapeutic Efficacy in Breast Cancer.

Front Med (Lausanne) 2021 9;8:667543. Epub 2021 Aug 9.

Department of Breast Surgery, First Affiliated Hospital of Army Military Medical University, Chongqing, China.

Increasing evidence highlights the roles of N-methyladenosine (mA) and its regulators in oncogenesis. Herein, this study observed the associations of mA regulators with breast cancer. RNA-seq profiles of breast cancer were retrieved from the Cancer Genome Atlas (TCGA) database. The expression of mA regulators was analyzed in tumor and normal tissues. Their expression correlations were analyzed by Spearson test. Overall survival (OS) analysis of these regulators was then presented. Gene set enrichment analysis (GSEA) was performed in high and low YTHDF1 expression groups. The correlations of YTHDF1 expression with immune cells and tumor mutation burden (TMB) were calculated in breast cancer samples. Somatic variation was assessed in high and low YTHDF1 expression groups. Most of mA regulators were abnormally expressed in breast cancer compared to normal tissues. At the mRNA levels, there were closely relationships between them. Among them, YTHDF1 up-regulation was significantly related to undesirable prognosis ( = 0.025). GSEA results showed that high YTHDF1 expression was associated with cancer-related pathways. Furthermore, YTHDF1 expression was significantly correlated with T cells CD4 memory activated, NK cells activated, monocytes, and macrophages. There were higher TMB scores in YTHDF1 up-regulation group than its down-regulation group. Missense mutation and non-sense mutation were the most frequent mutation types. Our findings suggested that dysregulated mA regulator YTHDF1 was predictive of survival outcomes as well as response to immunotherapy of breast cancer, and were closely related to immune microenvironment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fmed.2021.667543DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8380833PMC
August 2021

Isolation and Characterization of Shiga Toxin-Producing from Retail Beef Samples from Eight Provinces in China.

Foodborne Pathog Dis 2021 08;18(8):616-625

Department of Food Science, College of Food Science, Southwest University, Chongqing, China.

While Shiga toxin-producing (STEC) is a major foodborne pathogen worldwide, data on the molecular and phylogenetic properties of STEC isolates from retail beef samples in China remain scant. Fresh retail beef samples ( = 1062) were collected from eight provinces, and STEC isolates were recovered and characterized. PCR data showed that more than 50% of the samples were positive, and 82 STEC isolates were recovered from 14.8% (79/535) -positive enriched broths. In contrast, all ciprofloxacin resistant isolates ( = 19) and 13 cefotaxime (CTX) resistant isolates were positive and belonged to three serotypes: O111:H8, O26:H11, or O157:H7. Point mutations in quinolone resistance-determining regions and plasmid-mediated quinolone resistance determinants were identified in 16 and 20 isolates, respectively. and a point mutation (C-42T) in promoter were detected in 15 and 8 of the CTX resistant isolates, respectively. In addition, macrolide resistance gene was identified in eight azithromycin resistant O111:H8 isolates and one O26:H11 isolate. Single nucleotide polymorphism analysis demonstrated that the O26 and O157 isolates had multiple origins, but the O111 isolates were closely related. Taken together, our data demonstrated that several sequence types associated with hemolytic uremic syndrome from the retail beef samples in China had developed into dangerous multidrug resistant pathogens. The resistant phenotype can facilitate their transmission among the farm animals and human beings when there is an antimicrobial selective pressure.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/fpd.2021.0005DOI Listing
August 2021

Sevoflurane inhibits the apoptosis of hypoxia/reoxygenation-induced cardiomyocytes via regulating miR-27a-3p-mediated autophagy.

J Pharm Pharmacol 2021 Oct;73(11):1470-1479

Department of Anesthesiology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China.

Introduction: Sevoflurane (Sevo) prevents hypoxia/reoxygenation (H/R)-induced cardiomyocytes apoptosis. MiR-27a-3p expression is up-regulated in Sevo-treated hippocampal neurons.

Objective: This study explored whether the effect of Sevo on cardiomyocytes was mediated by miR-27a-3p.

Methods: The cardiomyocytes were cultured under H/R condition or pre-treated with Sevo, and further transfected with miR-27a-3p inhibitor or treated with an autophagy inhibitor 3-methyladenine (3-MA). Then the cell morphology was observed under an optical microscope. The cell viability and apoptosis were measured by MTT and flow cytometry. Expressions of miR-27a-3p, apoptosis-related, and autophagy-related factors were determined by western blot or RT-qPCR.

Key Findings: Sevo improved the abnormal morphology, promoted the cell viability and the expressions of Bcl-2 and miR-27a-3p, but reduced the apoptosis and Bax and C-caspase-3 levels of H/R-induced cardiomyocytes. MiR-27a-3p inhibitor had an effect opposite to Sevo on the cardiomyocytes and further counteracted the effect of Sevo on the H/R-induced cardiomyocytes. Downregulation miR-27a-3p increased the expression of Beclin 1 and the ratio of LC3B-II to LC3B-I in H/R-induced cardiomyocytes. Furthermore, 3-MA had an opposite effect to miR-27a-3p inhibitor and further counteracted the effect of the miR-27a-3p inhibitor on H/R-induced cardiomyocytes.

Conclusion: Sevo inhibited the apoptosis of H/R-induced cardiomyocytes via regulating miR-27a-3p-mediated autophagy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jpp/rgab111DOI Listing
October 2021

Automatic registration and precise tumour localization method for robot-assisted puncture procedure under inconsistent breath-holding conditions.

Int J Med Robot 2021 Aug 11:e2319. Epub 2021 Aug 11.

Department of Mechanical Engineering and Automation, Harbin Institute of Technology, Shenzhen, China.

Background: During percutaneous puncture procedure, breath holding is subjectively controlled by patients, and it is difficult to ensure consistent tumour position between the preoperative CT scanning phase and the intraoperative puncture phase. In addition, the manual registration process is time-consuming and has low accuracy.

Methods: We have proposed an automatic registration method using optical markers and a tumour breath-holding position estimation model based on the support vector regression algorithm. A robot system and a tumour respiratory motion simulation platform are built to perform puncture tests under different breath-holding states.

Results: The experimental results show that automatic registration has higher accuracy than manual registration, and with the tumour breath-holding position estimation model, the targeting accuracy of puncture under inconsistent breath-holding conditions is greatly improved.

Conclusions: The proposed automatic registration and tumour breath-holding position estimation model can improve the accuracy and efficiency of puncture under inconsistent breath-holding conditions.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/rcs.2319DOI Listing
August 2021
-->