Publications by authors named "Nan Zhang"

2,348 Publications

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Human endogenous retrovirus K (HERV-K) env in neuronal extracellular vesicles: a new biomarker of motor neuron disease.

Amyotroph Lateral Scler Frontotemporal Degener 2021 Jun 20:1-8. Epub 2021 Jun 20.

Department of Neurology, Peking University Third Hospital, Beijing, China.

Human endogenous retroviruses (HERVs) have been gradually confirmed to be involved in the onset and progression of motor neuron disease(MND). However, noninvasive detection of HERVs in the central nervous system is lacking. The aim of this study is to verify the relationship between the level of HERV-K env in neuronal extracellular vesicles in plasma and the onset and severity of MND. : We extracted neuronal extracellular vesicles from plasma of 39 MND patients and 30 age- and sex-matched controls, and detected HERV-K env in extracellular vesicles by an enzyme-linked immunosorbent assay (ELISA). : Levels of HERV-K env in neuronal extracellular vesicles positively associated with range of lower motor neurons (LMNs) involved (1.66 ± 0.37 vs. 1.35 ± 0.34,  = 0.041), ALS phenotype (1.52 ± 0.31 vs. 1.24 ± 0.37,  = 0.013) and course of disease (1.83 ± 0.35 vs. 1.42 ± 0.22,  = 0.003), and increased in advanced-phase MND (definite and probable according to revised EI Escorial criteria) compared with early-phase MND (possible and lab-supported probable), albeit without very profound significance (1.52 ± 0.34 vs. 1.29 ± 0.36,  = 0.048). : In conclusion, levels of HERV-K env in neuronal extracellular vesicles extracted from plasma can be used as a noninvasive biomarker of severity of MND.
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http://dx.doi.org/10.1080/21678421.2021.1936061DOI Listing
June 2021

An Open-Label, Randomized, 2-Way, Crossover Bioequivalence Study of Cefradine Capsules in Healthy Chinese Volunteers.

Clin Pharmacol Drug Dev 2021 Jun 20. Epub 2021 Jun 20.

Phase I Clinical Trial Center, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, China.

The purpose of this study was to evaluate whether test cefradine capsules and reference cefradine capsules were bioequivalent in healthy Chinese volunteers. An open-label, randomized, biperiodic, crossover design was used. In each of the 2 study periods (separated by a 1-week washout period), 250-mg single doses of either the test or reference cefradine capsule were administered to study participants under fasted and fed conditions. Blood samples were collected at intervals from predose to 8 hours afterward. In the fasting study, the 90% confidence intervals (90%CI) of the C , AUC , and AUC for the test and reference preparations were 93.7%-112.2%, 94.6%-100.8%, and 94.7%-100.9%, respectively. In the fed study, the 90%CI of the C , AUC , and AUC for the test and reference preparations was 81.0%-99.1%, 100.5%-106.3%, and 100.5%-105.9%, respectively. The results showed that the test cefradine capsules and the reference formulation are bioequivalent under both fasting and fed conditions.
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http://dx.doi.org/10.1002/cpdd.991DOI Listing
June 2021

A comprehensive prognostic signature for glioblastoma patients based on transcriptomics and single cell sequencing.

Cell Oncol (Dordr) 2021 Jun 17. Epub 2021 Jun 17.

One-third Lab, College of Bioinformatics Science and Technology, Harbin Medical University, Harbin, Hei Longjiang, 150000, People's Republic of China.

Purpose: Glioblastoma (GBM) is the most common and deadly brain tumor. We aimed to reveal potential prognostic GBM marker genes, elaborate their functions, and build an effective a prognostic model for GBM patients.

Methods: Through data mining of The Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA), we screened for significantly differentially expressed genes (DEGs) to calculate risk scores for individual patients. Published data of somatic mutation and copy number variation profiles were analyzed for distinct genomic alterations associated with risk scores. In addition, single-cell sequencing was used to explore the biological functions of the identified prognostic marker genes. By combining risk scores and other clinical features, we built a comprehensive prognostic GBM model.

Results: Seven DEGs (CLEC5A, HOXC6, HOXA5, CCL2, GPRASP1, BSCL2 and PTX3) were identified as being prognostic for GBM. Expression of these genes was confirmed in different GBM cell lines using real-time PCR. Risk scores calculated from the seven DEGs revealed prognostic value irrespective of other clinical factors, including IDH mutation status, and were negatively correlated with TP53 expression. The prognostic genes were found to be associated with tumor proliferation and progression based on pseudo-time analysis in neoplastic cells. A final prognostic model was developed and validated with a good performance, especially in geriatric GBM patients.

Conclusions: Using genetic profiles, age, IDH mutation status, and chemotherapy and radiotherapy, we constructed a comprehensive prognostic model for GBM patients. The model has a good performance, especially in geriatric GBM patients.
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http://dx.doi.org/10.1007/s13402-021-00612-1DOI Listing
June 2021

Diagnosis and treatment of pediatric anaplastic lymphoma kinase-positive large B-cell lymphoma: A case report.

World J Clin Cases 2021 Jun;9(17):4268-4278

Beijing Key Laboratory of Pediatric Hematology Oncology, National Discipline of Pediatrics, Ministry of Education, MOE Key Laboratory of Major Diseases in Children, Hematology Oncology Center, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing 100045, China.

Background: Anaplastic lymphoma kinase-positive (ALK+) large B-cell lymphoma (LBCL) is a rare type of lymphoma with high invasiveness and rapid progression. It occurs in all age groups, but is extremely rare in children. The lesions mainly involve the lymph nodes and may present with extra-nodal involvement. Response to conventional chemotherapies and local radiotherapy is poor, with a 5-year overall survival of less than 40%. Recently, the use of ALK inhibitors for the treatment of this disease has been reported.

Case Summary: We present a case of a 12-year-old boy diagnosed with ALK+LBCL. The patient had a 2-mo medical history of a calvarial mass, extensive systemic involvement, and positive bone marrow ( fusion gene. Complete remission 1 (CR1) was achieved using the modified LMB89 Group C regimen followed by autologous stem cell transplantation. The patient relapsed 3 mo later. He then achieved CR2 with three short courses of chemotherapy (COP, reduced-dose ICE, low-dose Ara-c+VP16) and continuous alectinib targeted therapy. Afterward, allogeneic hematopoietic stem cell transplantation (allo-HSCT) was performed. At 16 mo after the allo-HSCT, the patient was still in CR2.

Conclusion: The modified LMB89 Group C regimen and ALK inhibitors are effective. Allo-HSCT should be performed after remission.
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http://dx.doi.org/10.12998/wjcc.v9.i17.4268DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8173411PMC
June 2021

Omics-Based Identification of Shared and Gender Disparity Routes in -Induced Hepatocarcinogenesis: An Important Role for Dlk1-Dio3 Genomic Imprinting Region.

Front Genet 2021 31;12:620594. Epub 2021 May 31.

Department of Comparative Medicine, Laboratory Animal Center, Dalian Medical University, Dalian, China.

The phenomenon of gender disparity is very profound in hepatocellular carcinoma (HCC). Although previous research has revealed important roles of microRNA (miRNA) in HCC, there are no studies investigating the role of miRNAs in gender disparity observed hepatocarcinogenesis. In the present study, we investigated the global miRNAomics changes related to -induced male-prevalent hepatocarcinogenesis in a -transgenic mouse model (-Tg) by next-generation sequencing (NGS). We identified shared by also unique changes in miRNA expression profiles in gender-dependent hepatocarcinogenesis. Two hundred sixty-four differentially expressed miRNAs (DEMIRs) with value ≤0.05 and fold change ≥2 were identified. A vertical comparison revealed that the lower numbers of DEMIRs in the hepatic tumor (T) compared with the peri-tumor precancerous tissue (P) of -Tg and normal liver tissue of wild-type C57BL/6J mice (W) in males indicated that males are more susceptible to develop HCC. The expression pattern analysis revealed 43 common HCC-related miRNAs and 4 -positive-related miRNAs between males and females. By integrating the mRNA transcriptomic data and using 3-node FFL analysis, a group of significant components commonly contributing to HCC between sexes were filtered out. A horizontal comparison showed that the majority of DEMIRs are located in the Dlk1-Dio3 genomic imprinting region (GIR) and that they are closely related to not only hepatic tumorigenesis but also to gender disparity in hepatocarcinogenesis. This is achieved by regulating multiple metabolic pathways, including retinol, bile acid, and steroid hormones. In conclusion, the identification of shared and gender-dependent DEMIRs in hepatocarcinogenesis provides valuable insights into the mechanisms that contribute to male-biased -induced hepatic carcinogenesis.
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http://dx.doi.org/10.3389/fgene.2021.620594DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8202007PMC
May 2021

Effects of different parameters on the removal of atrazine in a water environment by biosorption.

J Pestic Sci 2021 May;46(2):214-221

Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.

Atrazine is the most extensively used herbicide in the agricultural and forestry sectors. Nevertheless, along with the increasing usage amount of Atrazine, its harm exposed gradually, the main problem is its residues in the environment. Microbial adsorption may effectively reduce the pollution caused by atrazine residue in the environment. In this study, a strain of fungi with the function of adsorbing atrazine was selected using microbial screening technology. According to its phenotypic characteristics and 18S rDNA gene sequencing, this strain was of the species genus and was named ECUST-TXZC2018. By studying the dynamic adsorption effect of this strain on atrazine, we found that this strain adsorbed atrazine after 36 hr at pH=5-7, and 20-30°C with more than 70% adsorption. These results demonstrated that ECUST-TXZC2018 had potential application ability to control atrazine residue pollution through the biosorption function.
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http://dx.doi.org/10.1584/jpestics.D20-043DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8175221PMC
May 2021

Quantitative analysis of retinal vessel density and thickness changes in diabetes mellitus evaluated using optical coherence tomography angiography: a cross-sectional study.

BMC Ophthalmol 2021 Jun 15;21(1):259. Epub 2021 Jun 15.

Eye Hospital, The First Affiliated Hospital of Harbin Medical University, Harbin, China.

Background: Diabetic retinopathy is the most common microvascular complication of diabetes; however, early changes in retinal microvessels are difficult to detect clinically, and a patient's vision may have begun to deteriorate by the time a problem is identified. Optical coherence tomography angiography (OCTA) is an innovative tool for observing capillaries in vivo. The aim of this study was to analyze retinal vessel density and thickness changes in patients with diabetes.

Methods: This was a retrospective, observational cross-sectional study. Between August 2018 and February 2019, we collected OCTA data from healthy participants and diabetics from the First Affiliated Hospital of Harbin Medical University. Analyzed their retinal vessel density and thickness changes.

Results: A total of 97 diabetic patients with diabetes at different severity stages of diabetic retinopathy and 85 controls were involved in the experiment. Diabetic patients exhibited significantly lower retinal VD (particularly in the deep vascular complexes), thickening of the neurosensory retina, and thinning of the retinal pigment epithelium compared with controls. In the control group, nondiabetic retinopathy group and mild diabetic retinopathy group, superficial VD was significantly correlated with retinal thickness (r = 0.3886, P < 0.0001; r = 0.3276, P = 0.0019; r = 0.4614, P = 0.0024, respectively).

Conclusions: Patients with diabetes exhibit ischemia of the retinal capillaries and morphologic changes in vivo prior to vision loss. Therefore, OCTA may be useful as a quantitative method for the early detection of diabetic retinopathy.
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http://dx.doi.org/10.1186/s12886-021-01988-2DOI Listing
June 2021

Dissecting the Flash Chemistry of Electrogenerated Reactive Intermediates by Microdroplet Fusion Mass Spectrometry.

Angew Chem Int Ed Engl 2021 Jun 15. Epub 2021 Jun 15.

Najing University, Chemistry, Xianlin Ave. 163, , 210023, Nanjing, CHINA.

A novel mass spectrometric method for probing the flash chemistry of electrogenerated reactive intermediates was developed based on rapid collision mixing of electrosprayed microdroplets by using a theta-glass capillary. The two individual microchannels of the theta-glass capillary are asymmetrically or symmetrically fabricated with a carbon bipolar electrode to produce intermediates in situ . Microdroplets containing the newly formed intermediates collide with those of the invoked reactants at sub-10 microsecond level, making it a powerful tool for exploring their ultrafast initial transformations. As a proof-of-concept, we present the identification of the key radical cation intermediate in the oxidative dimerization of 8-methyl-1,2,3,4-tetrahydroquinoline and also the first revealment of previously hidden nitrenium ion involved reaction pathway in the C-H/N-H cross-coupling between N,N'-dimethylaniline and phenothiazine.
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http://dx.doi.org/10.1002/anie.202106945DOI Listing
June 2021

PP04 improves high-fat diet-induced liver injury by the modulation of gut inflammation and intestinal microbiota in C57BL/6N mice.

Food Funct 2021 Jun 14. Epub 2021 Jun 14.

College of Food Science and Engineering, Jilin Agricultural University, Changchun, China. and National Engineering Laboratory for Wheat and Corn Deep Processing, Changchun, China.

In this study, Pediococcus pentococcus PP04 (PP04) isolated from the Northeast pickled cabbage was given to C57BL/6N mice for eight weeks, aiming to investigate the ameliorative effects of PP04 on liver injury induced by a high-fat diet. The western blot results suggested that PP04 ameliorated the increase of intestinal permeability by dramatically increasing the expressions of tight junction proteins, such as Occludin, Claudin-1 and ZO-1, which decreased hepatic lipopolysaccharides (LPS), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations to effectively alleviate the liver injury. Furthermore, PP04 relieved the high-fat diet-caused gut inflammation by the NF-κB/Nrf2 signaling pathway, which regulated the expression of inflammatory cytokines and antioxidants, to positively improve the liver injury. In addition, the 16S rDNA sequencing results inferred that PP04 had the potential to rebalance intestinal flora disorders through regulating the relative abundance of inflammation and obesity-related bacteria in mice.
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http://dx.doi.org/10.1039/d1fo00857aDOI Listing
June 2021

Comparison of decabromodiphenyl ether degradation in long-term operated anaerobic bioreactors under thermophilic and mesophilic conditions and the pathways involved.

J Environ Manage 2021 Jun 11;294:113009. Epub 2021 Jun 11.

Material Cycles Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8506, Japan; Fujian Ospring Technology Development Co., Ltd., No. 22 Jinrong North Road Cangshan District, Fuzhou, 350000, PR China. Electronic address:

Anaerobic digestion of decabromodiphenyl ether was carried out and compared in two continuously stirred anaerobic bioreactors for 210 days under thermophilic and mesophilic conditions. Results show that the degradation of decabromodiphenyl ether followed the first-order reaction kinetics, which exhibited a higher removal rate in the thermophilic reactor when compared to the mesophilic one, reaching its maximum of 1.1 μg·day. The anaerobic digestion of decabromodiphenyl ether was found to involve the replacement of bromines from polybrominated diphenyl ether by hydrogen atoms, gradually forming nona-, octa- and hepta-brominated diphenyl ether, respectively. Under the thermophilic condition, the reactors were dominated by Bacillus sp. and Methanosarcina sp. with high bioactivity and high concentrations of debromination microorganisms.
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http://dx.doi.org/10.1016/j.jenvman.2021.113009DOI Listing
June 2021

Examining the Effects of Brief Mindfulness Training on Athletes' Flow: The Mediating Role of Resilience.

Evid Based Complement Alternat Med 2021 24;2021:6633658. Epub 2021 May 24.

School of Psychology, Beijing Sport University, Beijing, China.

Background: Flow is characterized by the strong concentration in competitions, eliminating irrelevant thoughts and emotions, integrating all tasks, and continuing the competition smoothly even in challenging situations. The present study was into whether or not brief mindfulness training can improve athletes' flow and further explore the mediating effect of resilience in the intervention.

Methods: The 2 (experimental conditions) × 2 (time) mixed design was used in this study. Fifty-seven student-athletes were recruited and randomly assigned into either a brief mindfulness group ( = 29) or a control group ( = 28). Before and after the intervention, every participant completed a self-report measure including mindfulness, flow, and resilience.

Results: Participants in the brief mindfulness group showed increased mindfulness, flow, and resilience ( < 0.001) after brief mindfulness training; when putting resilience change ( = 0.30, 95% CI [0.031, 0.564]) into the equation, the direct (95% CI [3.156, 13.583]) and indirect (95% CI [0.470, 5.048]) effects of mindfulness training were both significant.

Conclusion: It was concluded that brief mindfulness training could significantly improve athletes' flow and resilience, and resilience partly mediated the effects of brief mindfulness training on flow.
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http://dx.doi.org/10.1155/2021/6633658DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8166472PMC
May 2021

A 4-aminonaphthalimide-based fluorescent traceable prodrug with excellent photoinduced cytotoxicity.

Chem Commun (Camb) 2021 Jun 11. Epub 2021 Jun 11.

Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, China. and University of Chinese Academy of Sciences, Beijing, 100049, China and School of Molecular Medicine, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, 310013, China.

A blue light activated anti-cancer prodrug, NST, was designed based on a photoactive 4-aminonaphthalimide derivative and an anticancer drug, 10-hydroxycamptothecin. NST was hard to be taken up by living cells and showed negligible dark cytotoxicity. The irradiation caused photocleavage of NST and resulted in high cytotoxicity.
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http://dx.doi.org/10.1039/d1cc01847gDOI Listing
June 2021

[Clinical effects of intermittent oro-esophageal tube feeding combined with Xuanqiaoliyan decotion in stroke patients with dysphagia].

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 2021 May;33(5):552-556

Department of Neurology, Yidu Central Hospital Affiliated Hospital of Weifang Medical University, Qingzhou 262500, Shandong, China. Corresponding author: Sun Xibo, Email:

Objective: To investigate the clinical effects of intermittent oro-esophageal tube feeding (IOE) combined with Xuanqiaoliyan decotion in stroke patients with dysphagia.

Methods: A prospective study was conducted. Stroke patients with dysphagia admitted to Yidu Central Hospital Affiliated Hospital of Weifang Medical University from January 2018 to December 2019 were enrolled. According to the simple random sampling method, the patients were divided into control group and observation group, with 50 cases in each group. The control group was given routine swallowing function training, including low-frequency pulse electrical stimulation, swallowing function training and acupuncture treatment. The observation group was given IOE and Xuanqiaoliyan decoction (prescription composition: Rhizoma acori tatarinowii 15 g, Radix polygalae 10 g, Rhizoma gastrodiae 15 g, Arisaema cum bile 6 g, Rhizoma typhonii 6 g, Scorpio 6 g, Bombyx batryticatus 6 g, Perilla frutescens 10 g, Rhizoma pinelliae 10 g, Pericarpium citri reticulatae 10 g, Rhizoma zingiberis recens 3 tablets, decoction 200 mL, twice in the morning and evening by oral or nasal feeding) on the basis of the control group. Both groups were treated for 14 days. The standard swallowing function assessment (SSA) and water swallow test were used to evaluate the swallowing function before and after treatment. The time required for the improvement of swallowing function, total hospitalization time and the therapeutic effects were observed and the safety assessment was conducted.

Results: There were no significant differences in the gender, age, course of disease, and location and frequency of stroke between the two groups. After treatment, both the SSA scores in the two groups were decreased, and the grading of water swallow test was improved. The SSA scores in the observation group were significantly lower than that in the control group (19.8±1.8 vs. 23.2±3.2, P < 0.05), the recovery degree of water swallow test was higher than that in the control group [complete recovery (cases): 18 vs. 13, basic recovery (cases): 23 vs. 18, effective (cases): 9 vs. 19, χ = -2.107, P = 0.008]. The total effective rate of swallowing function in the observation group was higher than that in the control group (94.0% vs. 80.0%, Z = 4.684, P = 0.012), the time for improvement (days: 12.8±2.6 vs. 16.9±4.3, t = 11.628, P = 0.008)and total hospitalization time (days: 20.8±4.2 vs. 33.5±5.6, t = 10.924, P = 0.015) were shorter than those in the control group. In the observation group, there was 1 case of throat discomfort during the operation of IOE, and the symptoms disappeared after the operation; there was 1 case of mild elevation of alanine aminotransferase (ALT) and blood urea nitrogen (BUN) respectively, which returned to normal after the treatment. No adverse symptoms and damage to the liver and kidney were observed in the control group.

Conclusions: IOE combined with Xuanqiaoliyan decotion could significantly improve the swallowing function of stroke patients with dysphagia, shorten the hospitalization time, and improve the curative effects and lifequality.
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http://dx.doi.org/10.3760/cma.j.cn121430-20200628-00487DOI Listing
May 2021

Highly Thermal-Wet Comfortable and Conformal Silk-Based Electrodes for On-Skin Sensors with Sweat Tolerance.

ACS Nano 2021 Jun 10. Epub 2021 Jun 10.

Innovative Centre for Flexible Devices (iFLEX), Max Planck-NTU Joint Lab for Artificial Senses, School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798 Singapore.

Noninvasive and seamless interfacing between the sensors and human skin is highly desired for wearable healthcare. Thin-film-based soft and stretchable sensors can to some extent form conformal contact with skin even under dynamic movements for high-fidelity signals acquisition. However, sweat accumulation underneath these sensors for long-term monitoring would compromise the thermal-wet comfort, electrode adherence to the skin, and signal fidelity. Here, we report the fabrication of a highly thermal-wet comfortable and conformal silk-based electrode, which can be used for on-skin electrophysiological measurement under sweaty conditions. It is realized through incorporating conducting polymers poly(3,4-ethylenedioxythiophene): polystyrenesulfonate (PEDOT:PSS) into glycerol-plasticized silk fiber mats. Glycerol plays the role of tuning the mechanical properties of silk fiber mats and enhancing the conductivity of PEDOT:PSS. Our silk-based electrodes show high stretchability (>250%), low thermal insulation (∼0.13 °C·m·W), low evaporative resistance (∼23 Pa·m·W, 10 times lower than ∼1.3 mm thick commercial gel electrodes), and high water-vapor transmission rate (∼117 g·m·h under sweaty conditions, 2 times higher than skin water loss). These features enable a better electrocardiography signal quality than that of commercial gel electrodes without disturbing the heat dissipation during sweat evaporation and provide possibilities for textile integration to monitor the muscle activities under large deformation. Our glycerol-plasticized silk-based electrodes possessing superior physiological comfortability may further engage progress in on-skin electronics with sweat tolerance.
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http://dx.doi.org/10.1021/acsnano.1c01431DOI Listing
June 2021

[Cognitive investigation and analysis of space loss in children].

Shanghai Kou Qiang Yi Xue 2021 Apr;30(2):177-181

Department of Stomatology, First Affiliated Hospital ,School of Medicine, Shihezi University. Shihezi 832008, Xinjiang Uygur Autonomous Region, China.

Purpose: To investigate the difference of parent's cognition of children's space loss and its influencing factors, to provide reference for children's oral health education, in order to reduce the incidence of malocclusion.

Methods: A self-designed questionnaire was used to investigate the basic information and related problems of 36 parents whose children received space loss treatment. SPSS 22.0 software package was used for statistical analysis.

Results: Parents paid more attention to the common oral problems of their children, the awareness of oral prevention and health care were weak, the attention to treatment was higher than that of prevention. The attitude of parents to the cognition and targeted treatment of malocclusion caused by space loss was low(27.78%), which was significantly different from that of caries (75.00%) and premature loss of deciduous teeth (63.89%). The total cognition of space loss was 61.11%. The cognitive degree of parents with different characteristics was different: the cognitive degree of parents over 50 years old was lower(14.29%), the cognitive degree of parents with higher education was higher(82.35%), there was no significant difference in cognitive degree among different gender, occupation and family monthly income groups. Parents paid more attention to the basic oral health of their children, most of the interval time between this visit and the last visit was 3 months(63.89%). The main purpose of the visit was caries or tooth pain caused by caries(63.89%). Most of parents did not know the harmful effect of space loss(72.22%). There were more parents who did not know the necessity of treatment to space loss(58.33%). Parents mainly learned the treatment to space loss through the doctor's education during dental visits(72.22%). The number of active visits were less when space loss occured(22.22%), and most of parents were recommended by dentist when they came to the clinics(72.22%). Most parents were able to accept the dentists' advice on the treatment of space loss(77.78%).

Conclusions: It is necessary to strengthen the propaganda, coverage and depth of oral health education care, strengthen the awareness of oral preventive health care, especially for the parents with special personality on the importance of space loss, to prevent primary teeth caries and premature loss of primary teeth, avoid the loss of dental space, make the complete primary dentition to form a normal permanent dentition.
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April 2021

Global profiling of RNA-binding protein target sites by LACE-seq.

Nat Cell Biol 2021 Jun 9;23(6):664-675. Epub 2021 Jun 9.

Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.

RNA-binding proteins (RBPs) have essential functions during germline and early embryo development. However, current methods are unable to identify the in vivo targets of a RBP in these low-abundance cells. Here, by coupling RBP-mediated reverse transcription termination with linear amplification of complementary DNA ends and sequencing, we present the LACE-seq method for identifying RBP-regulated RNA networks at or near the single-oocyte level. We determined the binding sites and regulatory mechanisms for several RBPs, including Argonaute 2 (Ago2), Mili, Ddx4 and Ptbp1, in mature mouse oocytes. Unexpectedly, transcriptomics and proteomics analysis of Ago2 oocytes revealed that Ago2 interacts with endogenous small interfering RNAs (endo-siRNAs) to repress mRNA translation globally. Furthermore, the Ago2 and endo-siRNA complexes fine-tune the transcriptome by slicing long terminal repeat retrotransposon-derived chimeric transcripts. The precise mapping of RBP-binding sites in low-input cells opens the door to studying the roles of RBPs in embryonic development and reproductive diseases.
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http://dx.doi.org/10.1038/s41556-021-00696-9DOI Listing
June 2021

Ultrasound-Guided Phrenic Nerve Block for Lung Nodule Biopsy: Single-Center Initial Experience.

Acad Radiol 2021 Jun 7. Epub 2021 Jun 7.

Department of Radiology, Mayo Clinic Hospital, Phoenix, Arizona, and Biostatistics, Mayo Clinic, Scottsdale, Arizona. Electronic address:

Rationale And Objectives: Biopsy of lung nodules in the lower lung fields can be difficult because of breathing motion. Ipsilateral phrenic nerve block (PNB) before biopsy should make the biopsy safer, easier, and more precisely targeted. We describe the use of ultrasound-guided PNB before lung nodule biopsy, including relevant anatomy and variations, complications, and technique, along with our first 40 cases.

Materials And Methods: We retrospectively reviewed patients who underwent PNB before computed tomography (CT)- or ultrasound-guided lung nodule biopsy from April 2015 through March 2020. Patient demographics, CT fluoroscopy time, radiation dose, complications, diagnostic yield, and effectiveness of PNB were recorded. Effectiveness of PNB was based on direct observation of diaphragmatic motion. Control group data for biopsies during the same time frame were collected and matched with nodules ≤1 cm from the PNB group.

Results: Among 40 patients identified, no complications occurred related to the PNB. Mean (SD) nodule size was 12.4 (6.2) mm. True-positive results were obtained in 39 patients (98%), with 1 false-negative after an ineffective PNB. PNB was effective in 70%. When CT fluoroscopy was used for the biopsy, radiation dose was significantly lower after an effective PNB than an ineffective PNB (p < .001). Effective PNB was significantly more common with injection of ≥4 mL of local anesthetic (p = .01). Comparison with 19 matched controls showed significantly fewer instances of pneumothorax (p = .02) and greater diagnostic success (p = .03) for the PNB group.

Conclusion: Ultrasound-guided PNB is safe and effective and can improve outcomes when used before lung nodule biopsy.
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http://dx.doi.org/10.1016/j.acra.2021.04.017DOI Listing
June 2021

In vivo sensing of rabbit cornea by terahertz technology.

J Biophotonics 2021 Jun 9:e202100130. Epub 2021 Jun 9.

Singapore Eye Research Institute (SERI), Singapore.

A Novel scalable approach using Terahertz (THz) waves together with the electromagnetic field simulation was applied to investigate four rabbits of eight rabbit corneas in vivo. One eye of each rabbits' corneas was edema induced; the other eye of the corneas served as the control. The simulation revealed the propagation of THz waves at a certain distance along the sub-surface of the cornea. THz spectra have been collected close to the corneal surface by deviating the direct reflection of the THz beam for the edema cornea, the reflected wave intensity for edema corneas is generally larger compared with the control cornea. Upon edema becomes severe at the end of the observation, the reflected wave intensities obtained by detector corresponding to the corneal deep stroma layer approach to the same value for all observed corneas. Good correlation is observed between central corneal thickness measurements and THz wave reflection signal intensities. Our results demonstrated that THz spectroscopy technique could obtain the information from different corneal sublayers.
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http://dx.doi.org/10.1002/jbio.202100130DOI Listing
June 2021

Age-Related Retinal Changes in Wild-Type C57BL/6J Mice Between 2 and 32 Months.

Invest Ophthalmol Vis Sci 2021 Jun;62(7)

Department of Ophthalmology, Emory University, Atlanta, Georgia, United States.

Purpose: The purpose of this study was to extend our understanding of how aging affects normal retina function and morphology in wild-type C57BL/6J mice, by analyzing electrophysiological recordings and in vivo and post mortem anatomy.

Methods: Electroretinograms (ERGs), spectral domain optical coherence tomography (SD-OCT), and confocal scanning laser ophthalmoscope (cSLO) in vivo images were obtained from mice between the ages of 2 and 32 months in four groups: group 1 (<0.5 years), group 2 (1.0-1.5 years), group 3 (1.5-2.0 years), and group 4 (>2.0 years). Afterward, mouse bodies and eyes were weighed. Eyes were stained with hematoxylin and eosin (H&E) and cell nuclei were quantified.

Results: With aging, mice showed a significant reduction in both a- and b-wave ERG amplitudes in scotopic and photopic conditions. Additionally, total retina and outer nuclear layer (ONL) thickness, as measured by SD-OCT images, were significantly reduced in older groups. The cSLO images showed an increase in auto-fluorescence at the photoreceptor-RPE interface as age increases. H&E cell nuclei quantification showed significant reduction in the ONL in older ages, but no differences in the inner nuclear layer (INL) or ganglion cell layer (GCL).

Conclusions: By using multiple age groups and extending the upper age limit of our animals to approximately 2.65 years (P970), we found that natural aging causes negative effects on retinal function and morphology in a gradual, rather than abrupt, process. Future studies should investigate the exact mechanisms that contribute to these gradual declines in order to discover pathways that could potentially serve as therapeutic targets.
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http://dx.doi.org/10.1167/iovs.62.7.9DOI Listing
June 2021

CRISPR/Cas9-mediated correction of homozygous point mutation in a Waardenburg syndrome 2A pig model.

Mol Ther Nucleic Acids 2021 Jun 16;24:986-999. Epub 2021 Apr 16.

State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.

Gene therapy for curing congenital human diseases is promising, but the feasibility and safety need to be further evaluated. In this study, based on a pig model that carries the c.740T>C (L247S) mutation in with an inheritance pattern and clinical pathology that mimics Waardenburg syndrome 2A (WS2A), we corrected the point mutation by the CRISPR-Cas9 system in the mutant fibroblast cells using single-stranded oligodeoxynucleotide (ssODN) and long donor plasmid DNA as the repair template. By using long donor DNA, precise correction of this point mutation was achieved. The corrected cells were then used as the donor cell for somatic cell nuclear transfer (SCNT) to produce piglets, which exhibited a successfully rescued phenotype of WS2A, including anophthalmia and hearing loss. Furthermore, engineered base editors (BEs) were exploited to make the correction in mutant porcine fibroblast cells and early embryos. The correction efficiency was greatly improved, whereas substantial off-targeting mutations were detected, raising a safety concern for their potential applications in gene therapy. Thus, we explored the possibility of precise correction of WS2A-causing gene mutation by the CRISPR-Cas9 system in a large-animal model, suggesting great prospects for its future applications in treating human genetic diseases.
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http://dx.doi.org/10.1016/j.omtn.2021.04.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8141604PMC
June 2021

Deciphering the mutational signature of congenital limb malformations.

Mol Ther Nucleic Acids 2021 Jun 20;24:961-970. Epub 2021 Apr 20.

Department of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China.

Congenital limb malformations (CLMs) affect 1 in 500 live births. However, the value of exome sequencing (ES) for CLM is lacking. The purpose of this study was to decipher the mutational signature of CLM on an exome level. We enrolled a cohort of 66 unrelated probands (including 47 families) with CLM requiring surgical correction. ES was performed for all patients and available parental samples. A definite molecular diagnosis was achieved in 21 out of 66 (32%) patients. We identified 19 pathogenic or likely pathogenic single-nucleotide variants and three copy number variants, of which 11 variants were novel. We identified four variants of uncertain significance. Additionally, we identified and as novel candidate genes for CLM. By comparing the detailed phenotypic features, we expand the phenotypic spectrum of diastrophic dysplasia and chromosome 6q terminal deletion syndrome. We also found that the diagnostic rate was significantly higher in patients with a family history of CLM (p = 0.012) or more than one limb affected (p = 0.034). Our study expands our understanding of the mutational and phenotypic spectrum of CLM and provides novel insights into the genetic basis of these syndromes.
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http://dx.doi.org/10.1016/j.omtn.2021.04.012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8141661PMC
June 2021

Regulatory mechanisms of immune checkpoints PD-L1 and CTLA-4 in cancer.

J Exp Clin Cancer Res 2021 Jun 4;40(1):184. Epub 2021 Jun 4.

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

The cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4)/B7 and programmed death 1 (PD-1)/ programmed cell death-ligand 1 (PD-L1) are two most representative immune checkpoint pathways, which negatively regulate T cell immune function during different phases of T-cell activation. Inhibitors targeting CTLA-4/B7 and PD1/PD-L1 pathways have revolutionized immunotherapies for numerous cancer types. Although the combined anti-CTLA-4/B7 and anti-PD1/PD-L1 therapy has demonstrated promising clinical efficacy, only a small percentage of patients receiving anti-CTLA-4/B7 or anti-PD1/PD-L1 therapy experienced prolonged survival. Regulation of the expression of PD-L1 and CTLA-4 significantly impacts the treatment effect. Understanding the in-depth mechanisms and interplays of PD-L1 and CTLA-4 could help identify patients with better immunotherapy responses and promote their clinical care. In this review, regulation of PD-L1 and CTLA-4 is discussed at the levels of DNA, RNA, and proteins, as well as indirect regulation of biomarkers, localization within the cell, and drugs. Specifically, some potential drugs have been developed to regulate PD-L1 and CTLA-4 expressions with high efficiency.
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http://dx.doi.org/10.1186/s13046-021-01987-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8178863PMC
June 2021

Proteomic Analysis Identifies Prolonged Disturbances in Pathways Related to Cholesterol Metabolism and Myocardium Function in the COVID-19 Recovery Stage.

J Proteome Res 2021 Jun 3. Epub 2021 Jun 3.

State Key Laboratory for Diagnosis and Treatment of Infectious Disease, National Clinical Research Center for Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.

The COVID-19 pandemic has become a worldwide health crisis. So far, most studies have focused on the epidemiology and pathogenesis of this infectious disease. Little attention has been given to the disease sequelae in patients recovering from COVID-19, and nothing is known about the mechanisms underlying these sequelae. Herein, we profiled the serum proteome of a cohort of COVID-19 patients in the disease onset and recovery stages. Based on the close integration of our proteomic analysis with clinical data, we propose that COVID-19 is associated with prolonged disorders in cholesterol metabolism and myocardium, even in the recovery stage. We identify potential biomarkers for these disorders. Moreover, severely affected patients presented more serious disturbances in these pathways. Our findings potentially support clinical decision-making to improve the prognosis and treatment of patients.
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http://dx.doi.org/10.1021/acs.jproteome.1c00054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8189036PMC
June 2021

Imaging features of mycotic aortic aneurysms.

Quant Imaging Med Surg 2021 Jun;11(6):2861-2878

Department of Cardiovascular Surgery, Beijing Aortic Disease Centre, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing Engineering Research Centre for Vascular Prostheses, Beijing, China.

Infectious aortitis (IA) is a rare and life-threatening cardiovascular disease. Early diagnosis and timely intervention are crucial for reducing mortality associated with mycotic aortic aneurysms (MAAs); however, early diagnosis is challenging due to the nonspecific symptoms. Some cases are diagnosed at an advanced stage or after developing complications, such as rupture or aortic fistula. Current state-of-the-art imaging modalities-including computed tomography (CT), magnetic resonance imaging (MRI), and 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET)/CT-can detect infected aneurysms in clinically suspicious cases. MAA features on imaging include lobulated pseudoaneurysm, indistinct irregular arterial wall, perianeurysmal gas, perianeurysmal edema, perianeurysmal soft tissue mass, aneurysmal thrombosis, and high metabolic activity with increased uptake of FDG. Enlarged lymph nodes are often found adjacent to the aneurysm, while iliopsoas abscess (IPA), spondylitis, and aortic fistulas are commonly associated complications. After surgery or endovascular repair, radiological features-including ectopic gas, peri-graft fluid, thickening of adjacent bowel, pseudoaneurysm formed at the graft anastomosis, and increased uptake of FDG-may indicate an infection of aortic graft. This article provides an overview of the clinical and imaging features of MAAs. Thus, familiarity with the imaging appearances of MAAs may assist radiologists in the diagnosis and facilitation of timely treatment.
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http://dx.doi.org/10.21037/qims-20-941DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8107294PMC
June 2021

Immunoregulation and antidepressant effect of ketamine.

Transl Neurosci 2021 Jan 26;12(1):218-236. Epub 2021 May 26.

Department of Psychiatry, Renmin Hospital of Wuhan University, Jiefang Rd. 238, 430060, Wuhan, China.

Major depressive disorder (MDD) is a common mental health disorder that brings severe disease burden worldwide. Traditional antidepressants are mainly targeted at monoamine neurotransmitters, with low remission rates and high recurrence rates. Ketamine is a noncompetitive glutamate -methyl-d-aspartate receptor (NMDAR) antagonist, and its rapid and powerful antidepressant effects have come to light. Its antidepressant mechanism is still unclarified. Research found that ketamine had not only antagonistic effect on NMDAR but also strong immunomodulatory effect, both of which were closely related to the pathophysiology of MDD. Although there are many related studies, they are relatively heterogeneous. Therefore, this review mainly describes the immune mechanisms involved in MDD and how ketamine plays an antidepressant role by regulating peripheral and central immune system, including peripheral inflammatory cytokines, central microglia, and astrocytes. This review summarizes the related research, finds out the deficiencies of current research, and provides ideas for future research and the development of novel antidepressants.
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http://dx.doi.org/10.1515/tnsci-2020-0167DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8155793PMC
January 2021

Effects of laparoscopic-assisted gastrectomy on elderly patients with gastric cancer.

J BUON 2021 Mar-Apr;26(2):490-498

Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Gastrointestinal Surgery IV, Peking University Cancer Hospital and Institute, Beijing, China.

Purpose: Although the acceptance of laparoscopy-assisted gastrectomy (LAG) for the treatment of gastric cancer (GC) has been increasing, it is still controversial that LAG is an applicable treatment method for elderly patients since elderly patients are usually complicated with other diseases. Therefore, this study aimed to investigate the prognostic differences between elderly patients and non-elderly patients after receiving LAG.

Methods: Patients (n = 306) who received LAG for the treatment of GC from April 2009 to December 2014 were included in the study. The patients were divided into the elderly group (≥ 65 years old, n = 120) and the non-elderly group (< 65 years old, n=186). The postoperative outcomes as well as the morbidity and the survival rates were compared between the two groups.

Results: American Society of Anesthesiologists (ASA) score and comprehensive complication index (CCI) score in the elderly group were significantly higher than those in the non-elderly group (p<0.05). In terms of surgical outcomes, there was no significant difference in blood loss or postoperative hospital stay between the elderly group and the non-elderly group. As for postoperative comorbidities, there were significant differences in intraperitoneal hemorrhage and pleural effusion between the elderly group and the non-elderly group. Moreover, the median follow-up time was 38.5 months, and the overall survival of elderly patients with comorbidities was significantly lower than that of the elderly patients without comorbidities (p<0.05).

Conclusions: LAG can be performed safely and successfully in the elderly population with acceptable postoperative and long-term results.
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June 2021

Tracking Changes in Age Distribution of Head and Neck Cancer in the U.S. from 1975-2016.

Clin Otolaryngol 2021 Jun 2. Epub 2021 Jun 2.

Division of Laryngology, Department of Otolaryngology - Head and Neck Surgery, Mayo Clinic Arizona, Phoenix, AZ, USA.

Introduction: Several independent studies report an alarming increase of patients younger than 40 being diagnosed with squamous cell carcinoma. There is currently a lack of available data clearly tracking changes in the age distribution of head and neck cancer (HNC) within the U.S. This study attempts to elucidate any trends in oral cavity, oropharynx, larynx and hypopharynx cancer age distribution in the United States population from 1975-2016. Unlike previous studies, this paper does not track incidence, but rather reports proportional changes of prevalence within age cohorts over time.

Methods: This is a retrospective chart review centered on data from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute (NCI). Each decade interval from 1975-2016 displays the proportion of HNC patients, classified by primary tumor subsite, within each age cohort.

Results: Mean age at diagnosis increased for all subsites except oropharynx. Oropharyngeal cancer was the only subsite to show a decrease in the mean age at diagnosis. In addition, oropharyngeal cancer was the only subsite to demonstrate an overall increase in proportional prevalence, largely due to increased incidence in middle age (40-59 years) patients. Cancers of the oral cavity were the only subset to show a true increase in the proportion of young (0-39 years) patients, but its mean age at diagnosis still increased. When stratifying by gender, the proportion of young patients in female HNC cases is higher than the young male proportion.

Conclusion: Overall, this study demonstrates an increased proportion of older HNC patients that is consistent with the aging population. Oral cavity cancer demonstrated a true increase in the proportion of young patients, likely due to the increased incidence of young women diagnosed with this cancer. Oropharyngeal cancer was the only subsite to show a decrease in the mean age at diagnosis. The increased proportion of middle-age patients with oropharyngeal cancer likely reflects the increase in HPV-related cancers.
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http://dx.doi.org/10.1111/coa.13817DOI Listing
June 2021

Screening of differential components of Gegenqinlian decoction and their comparative pharmacokinetics in normal and pyrexia rats by UHPLC-FT-ICR-MS and UHPLC-MS/MS.

Biomed Chromatogr 2021 Jun 2:e5186. Epub 2021 Jun 2.

School of Pharmacy, Shenyang Pharmaceutical University, Shenhe District, Shenyang, China.

An ultra-high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) metabonomic approach was employed to screen the differential components between normal rats and yeast-induced pyrexia rats after an oral administration of Gegenqinlian decoction (GQLD). Nine compounds, including puerarin, daidzein, baicalin, wogonoside, wogonin, berberine, palmatine, jateorhizine, and coptisine were identified as differential components in the plasma. A rapid, sensitive, selective, and accurate ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS) method was developed and fully validated for the simultaneous determination of the screened components in rat plasma after an oral administration of GQLD. The values for the limit of quantification ranged from 0.025 to 5.0 ng/ml. The inter- and intra-day precision of all analytes was ≤ 10.7%, with an accuracy of ≤ 10.5%. Good extraction recovery and matrix effects were also obtained. The method was successfully applied to a comparative pharmacokinetic study of GQLD in normal and pyrexia rats. The results showed that the pharmacokinetic behavior of the analytes was changed in pyrexia rats compared to normal rats. These results could provide beneficial guidance for clinical applications of GQLD.
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http://dx.doi.org/10.1002/bmc.5186DOI Listing
June 2021

Effect of Poly(L-lysine) and Heparin Coatings on the Surface of Polyester-Based Particles on Prednisolone Release and Biocompatibility.

Pharmaceutics 2021 May 27;13(6). Epub 2021 May 27.

Institute of Chemistry, St. Petersburg State University, Universitetskii Pr. 26, Peterhoff, 198504 St. Petersburg, Russia.

A plethora of micro- and nanoparticle types are currently investigated for advanced ocular treatment due to improved drug retention times, higher bioavailability and better biocompatibility. Yet, comparative studies of both physicochemical and toxicological performance of these novel drug delivery systems are still rare. Herein, poly(L-lactic acid)- and poly(ε-caprolactone)-based micro- and nanoparticles were loaded with prednisolone as a model drug. The physicochemical properties of the particles were varied with respect to their hydrophilicity and size as well as their charge and the effect on prednisolone release was evaluated. The particle biocompatibility was assessed by a two-tier testing strategy, combining the EpiOcular eye irritation test and bovine corneal opacity and permeability assay. The biodegradable polyelectrolyte corona on the particles' surface determined the surface charge and the release rate, enabling prednisolone release for at least 30 days. Thereby, the prednisolone release process was mainly governed by molecular diffusion. Finally, the developed particle formulations were found to be nontoxic in the tested range of concentrations.
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http://dx.doi.org/10.3390/pharmaceutics13060801DOI Listing
May 2021

A Single-Pass Type I Membrane Protein from the Apicomplexan Parasite with Nanomolar Binding Affinity to Host Cell Surface.

Microorganisms 2021 May 8;9(5). Epub 2021 May 8.

Key Laboratory for Zoonosis Research of the Ministry of Education, College of Veterinary Medicine, Institute of Zoonosis, Jilin University, Changchun 130062, China.

is a globally recognized zoonotic parasite of medical and veterinary importance. This parasite mainly infects intestinal epithelial cells and causes mild to severe watery diarrhea that could be deadly in patients with weakened or defect immunity. However, its molecular interactions with hosts and pathogenesis, an important part in adaptation of parasitic lifestyle, remain poorly understood. Here we report the identification and characterization of a T-cell immunomodulatory protein homolog (CpTIPH). CpTIPH is a 901-aa single-pass type I membrane protein encoded by cgd5_830 gene that also contains a short and (VCBS) repeat and relatively long integrin alpha (ITGA) N-terminus domain. Immunofluorescence assay confirmed the location of CpTIPH on the cell surface of sporozoites. In congruence with the presence of VCBS repeat and ITGA domain, CpTIPH displayed high, nanomolar binding affinity to host cell surface (i.e., at 16.2 to 44.7 nM on fixed HCT-8 and CHO-K1 cells, respectively). The involvement of CpTIPH in the parasite invasion is partly supported by experiments showing that an anti-CpTIPH antibody could partially block the invasion of sporozoites into host cells. These observations provide a strong basis for further investigation of the roles of CpTIPH in parasite-host cell interactions.
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http://dx.doi.org/10.3390/microorganisms9051015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8151451PMC
May 2021