Publications by authors named "Chaoqun Li"

80 Publications

Manganese-Catalyzed Achmatowicz Rearrangement Using Green Oxidant HO.

J Org Chem 2021 Jul 28;86(14):9563-9586. Epub 2021 Jun 28.

Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi'an 710062, China.

Oxidation reactions have been extensively studied in the context of the transformations of biomass-derived furans. However, in contrast to the vast literature on utilizing the stoichiometric oxidants, such as -CPBA and NBS, catalytic methods for the oxidative furan-recyclizations remain scarcely investigated. Given this, we report a means of manganese-catalyzed oxidations of furan with low loading, achieving the Achmatowicz rearrangement in the presence of hydrogen peroxide as an environmentally benign oxidant under mild conditions with wide functional group compatibility.
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http://dx.doi.org/10.1021/acs.joc.1c00858DOI Listing
July 2021

Nucleoside transporter-guided cytarabine-conjugated liposomes for intracellular methotrexate delivery and cooperative choriocarcinoma therapy.

J Nanobiotechnology 2021 Jun 15;19(1):184. Epub 2021 Jun 15.

Department of Pharmacy, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, 310006, China.

Gestational trophoblastic tumors seriously endanger child productive needs and the health of women in childbearing age. Nanodrug-based therapy mediated by transporters provides a novel strategy for the treatment of trophoblastic tumors. Focusing on the overexpression of human equilibrative nucleoside transporter 1 (ENT1) on the membrane of choriocarcinoma cells (JEG-3), cytarabine (Cy, a substrate of ENT1)-grafted liposomes (Cy-Lipo) were introduced for the targeted delivery of methotrexate ([email protected]) for choriocarcinoma therapy in this study. ENT1 has a high affinity for Cy-Lipo and can mediate the endocytosis of the designed nanovehicles into JEG-3 cells. The ENT1 protein maintains its transportation function through circulation and regeneration during endocytosis. Therefore, Cy-Lipo-based formulations showed high tumor accumulation and retention in biodistribution studies. More importantly, the designed DSPE-PEG-Cy conjugation exhibited a synergistic therapeutic effect on choriocarcinoma. Finally, [email protected] exerted an extremely powerful anti-choriocarcinoma effect with fewer side effects. This study suggests that the overexpressed ENT1 on choriocarcinoma cells holds great potential as a high-efficiency target for the rational design of active targeting nanotherapeutics.
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http://dx.doi.org/10.1186/s12951-021-00931-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8207694PMC
June 2021

Association between sleep disturbance and multidimensional frailty assessed by Frailty Index in hospitalized cirrhosis.

Eur J Gastroenterol Hepatol 2021 Jun 7. Epub 2021 Jun 7.

Department of Gastroenterology and Hepatology Tianjin Institute of Digestive Disease Department of Neurology, Tianjin Medical University General Hospital Department of Nutriology, Tianjin Third Central Hospital Department of Internal Medicine, Tianjin Hexi Hospital, Tianjin, China.

Objectives: Both sleep disturbance and frailty are common in patients with cirrhosis, but their correlation remains elusive. We aimed to investigate whether dysregulated sleep [as estimated by Pittsburgh Sleep Quality Index (PSQI)] is independently associated with frailty and their relationship in distinct subgroups.

Methods: In total 105 adult cirrhotic patients were recruited. The frailty phenotype was identified by a self-reported scale (Frailty Index) which demonstrates good validity and moderate performance based on our previous publication. Patients were categorized into frailty and nonfrailty groups according to a cut-point of 0.38 by Frailty Index. Multiple linear regression was performed to determine independent factors associated with frailty.

Results: The median PSQI was 6.0 in the entire cohort and sleep disturbance was observed in 61 patients with cirrhosis (58.1%). Poor sleepers had a significantly higher Frailty Index than that in good sleepers (0.11 vs. 0.08; P = 0.025). In univariate analysis, PSQI score was markedly associated with the Frailty Index (β = 0.012; 95% CI, 0.006-0.018; P < 0.001), and remained significantly associated with frailty phenotype in multivariate adjustment (β = 0.010; 95% CI, 0.004-0.015; P = 0.001). The escalating PSQI scores were more prominent in frail patients, with female gender or aged 65 years and over.

Conclusions: Poor sleep quality is strongly associated with frailty in patients with cirrhosis. Given that sleep disturbance is modifiable, our data suggest that efficient interventions to mitigate frailty should incorporate strategies by reversing sleep dysfunction in cirrhotics with poor sleep quality.
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http://dx.doi.org/10.1097/MEG.0000000000002231DOI Listing
June 2021

Learning From Crowds With Multiple Noisy Label Distribution Propagation.

IEEE Trans Neural Netw Learn Syst 2021 May 31;PP. Epub 2021 May 31.

Crowdsourcing services provide a fast, efficient, and cost-effective way to obtain large labeled data for supervised learning. Unfortunately, the quality of crowdsourced labels cannot satisfy the standards of practical applications. Ground-truth inference, simply called label integration, designs proper aggregation methods to infer the unknown true label of each instance (sample) from the multiple noisy label set provided by ordinary crowd labelers (workers). However, nearly all existing label integration methods focus solely on the multiple noisy label set per individual instance while totally ignoring the intercorrelation among multiple noisy label sets of different instances. To solve this problem, a multiple noisy label distribution propagation (MNLDP) method is proposed in this article. MNLDP at first estimates the multiple noisy label distribution of each instance from its multiple noisy label set and then propagates its multiple noisy label distribution to its nearest neighbors. Consequently, each instance absorbs a fraction of the multiple noisy label distributions from its nearest neighbors and yet simultaneously maintains a fraction of its own original multiple noisy label distribution. Empirical studies on a collection of an artificial dataset, six simulated UCI datasets, and three real-world crowdsourced datasets show that MNLDP outperforms all other existing state-of-the-art label integration methods in terms of the integration accuracy and classification accuracy.
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http://dx.doi.org/10.1109/TNNLS.2021.3082496DOI Listing
May 2021

Effect of urea feeding on transforming and migrating soil fluorine in a tea garden of hilly region.

Environ Geochem Health 2021 Apr 28. Epub 2021 Apr 28.

College of Chemistry and Materials Science, Sichuan Normal University, Chengdu, 610068, People's Republic of China.

The hilly region of western Sichuan is famous for producing Mengding-mountain tea in China. However, it is very easy to enrich fluorine (F) in tea, which recently raised people's concern on fluorosis. In this study, we simulated the local special climate conditions, especially the natural precipitation through column leaching experiment to investigate the effect of fertilization on F desorption from soil. The results suggested different fertilizers could greatly affect desorption of F from soil, but only urea (CO(NH)) could significantly and continuously promote the dissolution of F from soil. Furthermore, to reveal the effect of CO(NH) on migration of F from soil to tea, field plots experiment in a tea garden was carried out. The results showed that CO(NH) promoted soil acidification continuously, resulting in the increase of dissolved F and Al content in soil, which were migrated to tea in the form of F-Al complex. Therefore, the key to reducing the F content in tea is to alleviate soil acidification and reduce F-Al complex content in soil. Most importantly, with CO(NH) addition, the contents of F in tea increased significantly from first bud to fifth bud in comparison with those of without CO(NH), especially in 2016, indicating the accelerating and prolonging enrichment of F in tea by adding CO(NH). This study would have important guidance for controlling F mitigation from soil to tea by managing fertilizers application and anthropogenic activities.
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http://dx.doi.org/10.1007/s10653-021-00949-4DOI Listing
April 2021

Abiotic and biotic factors contribute to CO exchange variation at the hourly scale in a semiarid maize cropland.

Sci Total Environ 2021 Aug 18;784:147170. Epub 2021 Apr 18.

College of Mechanical and Electronic Engineering, Northwest A&F University, Yangling, Shaanxi 712100, China; Key Laboratory of Agricultural of Things, Ministry of Agriculture, Yangling, China.

Understanding the variables influencing the carbon budget in agricultural ecosystems is crucial for the prediction of future carbon dynamics. The purpose of this study was to identify the biotic and abiotic determinants of the net ecosystem CO exchange (NEE) and net assimilation rate (NPP) in a semiarid maize cropland. The CO exchange (NEE and NPP) was measured at different growth stages of maize plants using an improved chamber methodology. Heat map clustering of the correlation coefficients between CO exchange and its driving factors demonstrated that soil temperature and air humidity were positively correlated with CO emissions regardless of daytime or nighttime, while other factors affecting CO exchange were negatively correlated with emissions during daytime yet positively correlated during nighttime. The machine learning algorithm random forest (RF) and structural equation modeling (SEM) were used to analyze the effects of different factors on CO exchange. The RF analysis results indicated that for CO exchange in the daytime, photosynthetically active radiation (PAR) was the most important variable and presented an importance score of 0.574 for NEE and 0.558 for NPP. The SEM results indicated that in the daytime PAR exerted significant direct and indirect effects on both NEE and NPP, and the standardized direct and indirect effects were -0.668 and 0.022, respectively, for NEE, and the effects were 0.655 and -0.011, respectively for NPP. Like PAR, soil water content also exerted significant direct and indirect effects on both NEE and NPP, but the remaining factors affecting CO exchange only have one of the direct or indirect effects, sometimes neither. For CO exchange at night, the leaf area was the most important variable and presented an importance score of 0.72 for NEE and 0.45 for NPP. At night, both the direct and indirect effects of most abiotic factors on NEE and NPP were significant.
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http://dx.doi.org/10.1016/j.scitotenv.2021.147170DOI Listing
August 2021

Angiopep-2-modified calcium arsenite-loaded liposomes for targeted and pH-responsive delivery for anti-glioma therapy.

Biochem Biophys Res Commun 2021 04 11;551:14-20. Epub 2021 Mar 11.

College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China. Electronic address:

The blood-brain barrier (BBB) is the most critical obstacle in the treatment of central nervous system disorders, such as glioma, the most typical type of brain tumor. To overcome the BBB and enhance drug-penetration abilities, we used angiopep-2-modified liposomes to deliver arsenic trioxide (ATO) across the BBB, targeting the glioma. Angiopep-2-modified calcium arsenite-loaded liposomes ([email protected]), with uniformly distributed hydrodynamic diameter (96.75 ± 0.57 nm), were prepared using the acetate gradient method with high drug-loading capacity (7.13 ± 0.72%) and entrapment efficiency (54.30 ± 9.81%). In the acid tumor microenvironment, arsenic was responsively released, thereby exerting an anti-glioma effect. The anti-glioma effect of [email protected] was investigated both in vitro and in vivo. As a result, [email protected] exhibited a potent, targeted anti-glioma effect mediated by the lipoprotein receptor-related (LRP) receptor, which is overexpressed in both the BBB and glioma. Therefore, [email protected] could dramatically promote the anti-glioma effect of ATO, as a promising strategy for glioma therapy.
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http://dx.doi.org/10.1016/j.bbrc.2021.02.138DOI Listing
April 2021

Long noncoding RNA MCM3AP-AS1 enhances cell proliferation and metastasis in colorectal cancer by regulating miR-193a-5p/SENP1.

Cancer Med 2021 04 8;10(7):2470-2481. Epub 2021 Mar 8.

Wuxi Cancer Institute, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China.

Background: Accumulating evidences have shown that long noncoding RNAs (lncRNAs) play key roles in many diseases, including cancer. Several studies reported that MCM3AP antisense RNA 1 (MCM3AP-AS1) was associated with the tumorigenesis and progression. However, the specific function and mechanism of MCM3AP-AS1 in colorectal cancer (CRC) have not been fully understood.

Methods: The expression of MCM3AP-AS1 was detected by quantitative reverse transcription PCR (RT-qPCR) in CRC tissues and matched noncancerous tissues (NCTs). CCK-8 assay, colony formation assay, transwell assay, xenograft and lung metastasis mouse models were used to examine the tumor-promoting function of MCM3AP-AS1 in vitro and in vivo. The binding relationship between MCM3AP-AS1, miR-193a-5p and sentrin-specific peptidase 1 (SENP1) were screened and identified by databases, RT-qPCR, dual luciferase reporter assay and western blot.

Results: In the present study, we got that the expression of MCM3AP-AS1 was higher in CRC tissues than in paired NCTs, and increased MCM3AP-AS1 expression was associated with adverse outcomes in CRC patients. Functional experiments in vitro revealed that silencing of MCM3AP-AS1 could inhibit the proliferation, colony formation, migratory, and invasive abilities of CRC cells. The mouse models of xenograft and lung metastasis further confirmed that in vivo silencing MCM3AP-AS1 could significantly inhibit the growth and metastasis of CRC. Further mechanism studies indicated that MCM3AP-AS1 could sponge miR-193a-5p and inhibit the activity of it. What is more, SENP1 was proved to be a novel target of miR-193a-5p and could be upregulated by MCM3AP-AS1. At last, we observed that SENP1 overexpression in CRC tissues was closely related to unfavorable prognosis.

Conclusion: Taken together, we identified in CRC the MCM3AP-AS1/miR-193a-5p/SENP1 regulatory axis, which affords a therapeutic possibility for CRC.
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http://dx.doi.org/10.1002/cam4.3830DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7982620PMC
April 2021

Relationship between work stressors and mental health in frontline nurses exposed to COVID-19: A structural equation model analysis.

Ann Med Psychol (Paris) 2021 Feb 16. Epub 2021 Feb 16.

Department of Nursing, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, China.

The current cross-sectional study aimed to explore the relationship between work stressors and mental health in frontline nurses exposed to COVID-19. Participants were recruited from 16 general hospitals in Anhui province from February 2020 to March 2020. The general sociodemographic questionnaire, Nurse Job Stressors Scale, Simplified Coping Style Questionnaire, NEO Five-Factor Inventory, Perceived Social Support Scale, and Kessler Psychological Distress Scale were used in this study. Based on 723 valid questionnaires retrieved (100%), the total mean scores of work stressors and mental health of frontline nurses were (94.38±23.42) and (22.81±7.16), respectively. The results of the structural equation model showed that work stressors had an indirect positive effect (β=0.484, P<0.01), social support had a direct negative effect (β=-0.934, P<0.01), personality traits had a direct positive effect (β=0.209, P<0.01), and positive coping style had both direct positive (β=0.246, P<0.01) and indirect negative effects (β=-0.873, P<0.01) on frontline nurses' mental health. In conclusion, nursing staff can reinforce positive influences by accepting social support, adopting positive coping methods, and weakening negative influences factors to reduce or buffer their negative mental states and further reduce work stress.
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http://dx.doi.org/10.1016/j.amp.2021.02.015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7885667PMC
February 2021

Different Crystal Forms of ZnS Nanomaterials for the Adsorption of Elemental Mercury.

Environ Sci Technol 2021 05 6;55(10):6965-6974. Epub 2021 Feb 6.

CAS Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Innovation Academy for Green Manufacture, Chinese Academy of Sciences, Beijing 100190, P. R. China.

ZnS is a promising sorbent in recovering Hg from industrial flue gas due to its excellent Hg adsorption capacity. However, the internal structure-activity relationship still needs to be further clarified. In this work, ZnS sorbents with different structures were synthesized with the hydrothermal method by tuning the temperature. The samples had significant differences in the crystallinity, morphology, particle size, and sulfur (S) active sites. The results indicated that Hg removal performance was determined by the specific surface area and S active sites. ZnS synthesized at low temperatures (80-ZnS and 120-ZnS) had a larger surface area, while the S sites on the high-temperature-synthesized sample (160-ZnS) were more active for Hg adsorption. The 160-ZnS sample exhibited a much higher Hg adsorption amount per unit surface area. Further characterization revealed that S and S were the main active sites for Hg adsorption. S existed in the form of long-chain polysulfur (L-S) on 80-ZnS and 120-ZnS, while it exhibited in the form of short-chain polysulfur (S-S) on 160-ZnS. L-S had negligible adsorption ability, while S-S had a high affinity for Hg. Hg can react with S and S-S, forming α-HgS and β-HgS, respectively. The new insight in this work can provide theoretical guidance for the design and structure optimization of ZnS, facilitating its practical industrial application.
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http://dx.doi.org/10.1021/acs.est.0c05878DOI Listing
May 2021

Enantioconvergent Amination of Racemic Tertiary C-H Bonds.

J Am Chem Soc 2020 12 29;142(49):20902-20911. Epub 2020 Nov 29.

Department of Chemistry, Merkert Chemistry Center, Boston College, Chestnut Hill, Massachusetts 02467, United States.

Racemization is considered to be an intrinsic stereochemical feature of free radical chemistry as can be seen in traditional radical halogenation reactions of optically active tertiary C-H bonds. If the facile process of radical racemization could be effectively combined with an ensuing step of bond formation in an enantioselective fashion, then it would give rise to deracemizative functionalization of racemic tertiary C-H bonds for stereoselective construction of chiral molecules bearing quaternary stereocenters. As a demonstration of this unique potential in radical chemistry, we herein report that metalloradical catalysis can be successfully applied to devise Co(II)-based catalytic system for enantioconvergent radical amination of racemic tertiary C(sp)-H bonds. The key to the success of the radical process is the development of Co(II)-based metalloradical catalyst with fitting steric, electronic, and chiral environments of the -symmetric chiral amidoporphyrin as the supporting ligand. The existence of optimal reaction temperature is recognized as an important factor in the realization of the enantioconvergent radical process. Supported by an optimized chiral ligand, the Co(II)-based metalloradical system can effectively catalyze the enantioconvergent 1,6-amination of racemic tertiary C(sp)-H bonds at the optimal temperature, affording chiral α-tertiary amines in excellent yields with high enantiocontrol of the newly created quaternary stereocenters. Systematic studies, including experiments utilizing optically active deuterium-labeled C-H substrates as a model system, shed light on the underlying mechanistic details of this new catalytic process for enantioconvergent radical C-H amination. The remarkable power to create quaternary stereocenters bearing multiple functionalities from ubiquitous C-H bonds, as showcased with stereoselective construction of bicyclic -heterocycles, opens the door for future synthetic applications of this new radical technology.
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http://dx.doi.org/10.1021/jacs.0c11103DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7725894PMC
December 2020

Effect of pharmaceutical care on the treatment of COVID-19: A protocol for systematic review and meta analysis.

Medicine (Baltimore) 2020 Nov;99(48):e23377

Department of Pharmacy, Jingjiang People's Hospital, the Seventh Affiliated Hospital of Yangzhou University, Jingjiang, Jiangsu, China.

Background: We aimed to conduct a meta-analysis to assess the effect of pharmaceutical care on the treatment of coronavirus disease 2019 (COVID-19).

Methods: All case-controlled studies related to pharmaceutical care on the treatment of COVID-19 will be included in this review. We will use index words related to pharmaceutical care and COVID-19 to perform literature searches in PubMed, Embase, MEDLINE, CNKI, and Wanfang databases, to include articles indexed as of October 20, 2020 in English and Chinese language. Two reviewers will select trials independently for inclusion and assess trial quality. Two pairs of review authors will independently extract information for each included trials. Primary outcomes are clinical outcomes, average hospital stays, costs, patient satisfaction, and incidence of adverse drug reactions. We will evaluate the risk of bias of the included studies based on Cochrane assessment tool. Revman 5.3 (the Cochrane collaboration, Oxford, UK) will be used for heterogeneity assessment, generating funnel-plots, data synthesis, subgroup analysis, and sensitivity analysis.

Results: We will provide targeted and practical results assessing the effect of pharmaceutical care on the treatment of COVID-19.

Conclusion: The stronger evidence about the effect of pharmaceutical care on the treatment of COVID-19 will be provided for clinicians.

Systematic Review Registration Number: PROSPERO CRD42020214223 ETHICS AND DISSEMINATION:: There is no need for ethical approval, and the review will be reported in a peer-reviewed journal.
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http://dx.doi.org/10.1097/MD.0000000000023377DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7710203PMC
November 2020

Bioactive metal-containing nanomaterials for ferroptotic cancer therapy.

J Mater Chem B 2020 12;8(46):10461-10473

Department of Pharmacy, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.

The clinical performance of the current cancer therapies is still far from satisfactory. The emerging ferroptosis-driven therapy strategies reignite the hope of chemotherapy in tumor treatment due to their incredible tumor suppression. Among ferroptosis-based cancer therapies, metal elements have attracted remarkable attention due to their inherent physicochemical properties in inducing ferroptosis of tumor cells quickly and strongly without complex cellular signal transduction. Although the discovery and applications of ferroptosis for tumor treatment have been discussed in many reviews, the unique advantages of metal-containing nanomaterials interfering ferroptotic cancer therapies (MIFCT) have seldom been mentioned. Here, we outline the latest advances of MIFCT comprehensively. Firstly, the functions of different kinds of metal elements or their ions are introduced to illustrate their advantages in MIFCT. Secondly, the emerging metal-containing nanomaterials that are designed to achieve ferroptosis-driven therapy are overviewed, including their ability to boost the Fenton or Fenton-like reaction for reactive oxygen species generation, act as hydrogen peroxide self-providers, damage the reducing system, and disturb cellular communication. Moreover, metal-containing nanomaterials with external energy conversion features for MIFCT are discussed. Finally, the future expectations and challenges of MIFCT for clinical cancer therapy are spotlighted.
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http://dx.doi.org/10.1039/d0tb02138eDOI Listing
December 2020

Developing an Automated Gas Sampling Chamber for Measuring Variations in CO2 Exchange in a Maize Ecosystem at Night.

Sensors (Basel) 2020 Oct 27;20(21). Epub 2020 Oct 27.

College of Mechanical and Electronic Engineering, Northwest A&F University, Yangling 712100, China.

The measurement of net ecosystem exchange (NEE) of field maize at a plot-sized scale is of great significance for assessing carbon emissions. Chamber methods remain the sole approach for measuring NEE at a plot-sized scale. However, traditional chamber methods are disadvantaged by their high labor intensity, significant resultant changes in microclimate, and significant impact on the physiology of crops. Therefore, an automated portable chamber with an air humidity control system to determinate the nighttime variation of NEE in field maize was developed. The chamber system can automatically open and close the chamber, and regularly collect gas in the chamber for laboratory analysis. Furthermore, a humidity control system was created to control the air humidity of the chamber. Chamber performance test results show that the maximum difference between the temperature and humidity outside and inside the chamber was 0.457 °C and 5.6%, respectively, during the NEE measuring period. Inside the chamber, the leaf temperature fluctuation range and the maximum relative change of the maize leaf respiration rate were 0.3 to 0.3 °C and 23.2015%, respectively. We verified a series of measurements of NEE using the dynamic and static closed chamber methods. The results show a good common point between the two measurement methods (N = 10, R = 0.986; and mean difference: △CO = 0.079  ). This automated chamber was found to be useful for reducing the labor requirement and improving the time resolution of NEE monitoring. In the future, the relationship between the humidity control system and chamber volume can be studied to control the microclimate change more accurately.
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http://dx.doi.org/10.3390/s20216117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7662807PMC
October 2020

Mathematical models for devising the optimal SARS-CoV-2 strategy for eradication in China, South Korea, and Italy.

J Transl Med 2020 09 5;18(1):345. Epub 2020 Sep 5.

Shanghai Public Health Clinical Center, Fudan University, 2901 Caolang Road, Jinshan District, Shanghai, 201508, People's Republic of China.

Background: Coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spreads rapidly and has attracted worldwide attention.

Methods: To improve the forecast accuracy and investigate the spread of SARS-CoV-2, we constructed four mathematical models to numerically estimate the spread of SARS-CoV-2 and the efficacy of eradication strategies.

Results: Using the Susceptible-Exposed-Infected-Removed (SEIR) model, and including measures such as city closures and extended leave policies implemented by the Chinese government that effectively reduced the β value, we estimated that the β value and basic transmission number, R, of SARS-CoV-2 was 0.476/6.66 in Wuhan, 0.359/5.03 in Korea, and 0.400/5.60 in Italy. Considering medicine and vaccines, an advanced model demonstrated that the emergence of vaccines would greatly slow the spread of the virus. Our model predicted that 100,000 people would become infected assuming that the isolation rate α in Wuhan was 0.30. If quarantine measures were taken from March 10, 2020, and the quarantine rate of α was also 0.3, then the final number of infected people was predicted to be 11,426 in South Korea and 147,142 in Italy.

Conclusions: Our mathematical models indicate that SARS-CoV-2 eradication depends on systematic planning, effective hospital isolation, and SARS-CoV-2 vaccination, and some measures including city closures and leave policies should be implemented to ensure SARS-CoV-2 eradication.
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http://dx.doi.org/10.1186/s12967-020-02513-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7474336PMC
September 2020

Recent Advances in the Development of Virus-Like Particle-Based Flavivirus Vaccines.

Vaccines (Basel) 2020 Aug 27;8(3). Epub 2020 Aug 27.

Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY 10065, USA.

Flaviviruses include several medically important viruses, such as Zika virus (ZIKV), Dengue virus (DENV), West Nile virus (WNV) and Japanese encephalitis virus (JEV). They have expanded in geographic distribution and refocused international attention in recent years. Vaccination is one of the most effective public health strategies for combating flavivirus infections. In this review, we summarized virus-like particle (VLP)-based vaccines against the above four mentioned flaviviruses. Potential strategies to improve the efficacy of VLP-based flavivirus vaccines were also illustrated. The applications of flavivirus VLPs as tools for viral detection and antiviral drug screening were finally proposed.
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http://dx.doi.org/10.3390/vaccines8030481DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7565697PMC
August 2020

Targeted Manganese doped silica nano GSH-cleaner for treatment of Liver Cancer by destroying the intracellular redox homeostasis.

Theranostics 2020 2;10(21):9865-9887. Epub 2020 Aug 2.

College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 311400, China.

Glutathione (GSH), the primary antioxidant in cells, could fight against oxidative stress. Tumor cells display a higher GSH level than normal cells for coping with the hyperoxidative state, which meets the requirements of enhanced metabolism and vicious proliferation. Therefore, the consumption of GSH will lead to cell redox imbalance and impede life activities. Herein, targeted sorafenib (SFB) loaded manganese doped silica nanoparticle ([email protected]) was constructed, which could destroy the intracellular redox homeostasis by consuming GSH. In this study, MnMSN was prepared by an optimized one-pot Stober's method for loading SFB, and FaPEG chain was modified on the surface of MnMSN to achieve long circulation and targeted delivery. The anticancer efficacy and mechanism of the designed [email protected] were assessed both and [email protected] exhibited efficient antitumor activity by dual depleting intracellular GSH (the degradation of MnMSN would consume intracellular GSH and the SFB would inhibit the effect of X transport system to inhibit GSH synthesis). Moreover, disruption of redox balance would lead to apoptosis and reactive oxygen species (ROS)-dependent ferroptosis of tumor cells. Such a GSH-starvation therapeutic strategy would cause multi-path programmed cell death and could be a promising strategy for cancer therapy.
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http://dx.doi.org/10.7150/thno.46771DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7449918PMC
May 2021

An overview of Middle East respiratory syndrome coronavirus vaccines in preclinical studies.

Expert Rev Vaccines 2020 09 8;19(9):817-829. Epub 2020 Sep 8.

Lindsley F. Kimball Research Institute, New York Blood Center , New York, NY, USA.

Introduction: Middle East respiratory syndrome coronavirus (MERS-CoV) causes high mortality in humans. No vaccines are approved for use in humans; therefore, a consistent effort to develop safe and effective MERS vaccines is needed.

Areas Covered: This review describes the structure of MERS-CoV and the function of its proteins, summarizes MERS vaccine candidates under preclinical study (based on spike and non-spike structural proteins, inactivated virus, and live-attenuated virus), and highlights potential problems that could prevent these vaccines entering clinical trials. It provides guidance for the development of safe and effective MERS-CoV vaccines.

Expert Opinion: Although many MERS-CoV vaccines have been developed, most remain at the preclinical stage. Some vaccines demonstrate immunogenicity and efficacy in animal models, while others have potential adverse effects or low efficacy against high-dose or divergent virus strains. Novel strategies are needed to design safe and effective MERS vaccines to induce broad-spectrum immune responses and improve protective efficacy against multiple strains of MERS-CoV and MERS-like coronaviruses with pandemic potential. More funds should be invested to move vaccine candidates into human clinical trials.
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http://dx.doi.org/10.1080/14760584.2020.1813574DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7755455PMC
September 2020

Targeted GSH-exhausting and hydroxyl radical self-producing manganese-silica nanomissiles for MRI guided ferroptotic cancer therapy.

Nanoscale 2020 Aug;12(32):16738-16754

Department of Pharmacy, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.

Ferroptosis, a cell death path induced by the generation of reactive oxygen species (ROS), will cause the accumulation of lipid peroxides (PL-PUFA-OOH) and achieve potent tumor-regression. However, glutathione (GSH)-dependent glutathione peroxidase 4 (GPx4) can reduce PL-PUFA-OOH and antagonize the ferroptosis inducing effect of ROS. Herein, folate-PEG modified dihydroartemisinin (DHA) loaded manganese doped mesoporous silica nanoparticles (described as nanomissiles) were constructed for integrating the effect of GSH exhaustion and ROS generation. After endocytosis by tumor cells, intracellular GSH triggered the degradation of nanomissiles, which allowed the simultaneous release of DHA and Fenton catalytic Mn2+ due to the redox reaction between the manganese-oxygen bonds and GSH. The degradation would lead to GSH exhaustion, activation of Mn2+-based magnetic resonance imaging (MRI), and DHA-driven ˙OH generation. The GSH-free environment inhibited the activity of GPx4 and enhanced the accumulation of PL-PUFA-OOH oxidized by ˙OH. Furthermore, the cooperative effects suppressed tumor metastasis by destroying the structure of polyunsaturated fatty acids in the cell membranes and showed potent antitumor activity. This innovative ferroptotic therapy integrating the GSH exhaustion and ROS generation will be a promising strategy for cancer therapy.
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http://dx.doi.org/10.1039/d0nr02396eDOI Listing
August 2020

Administration of commensal Shewanella sp. MR-7 ameliorates lipopolysaccharide-induced intestine dysfunction in turbot (Scophthalmus maximus L.).

Fish Shellfish Immunol 2020 Jul 7;102:460-468. Epub 2020 May 7.

Key Laboratory of Mariculture (Ministry of Education), Ocean University of China, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China. Electronic address:

This study was designed to evaluate whether the administration of commensal Shewanella sp. MR-7 (MR-7) could ameliorate lipopolysaccharide (LPS)-induced intestine dysfunction in turbot. Fish (body weight: 70.00 ± 2.00 g) were randomly divided into three groups including the control group treated with dough, the LPS group treated with dough plus LPS, and the LPS+MR-7 (LMR) group treated with dough plus LPS and MR-7. These three groups with 24 fish each were force-fed with 1 g dough daily for 7 continuous days. The results revealed that MR-7 administration ameliorated LPS-induced intestinal injury, showing higher intestinal villus and microvillus height. Further results showed that MR-7 could inhibit LPS-induced activation of TLR-NF-κB signaling thus maintaining the normal expression levels of cytokines and finally ameliorate the intestinal inflammatory response in turbot. Compared with the LPS group, LMR group had less goblet cells and lower mucin-2 expression level. Moreover, MR-7 restored LPS-induced down-regulation of tight junction protein-related gene expression (zonula occluden-1, occludin, tricellulin and claudin-3). Further investigations indicated that MR-7 partially counteracted LPS-induced changes in gut microbiota composition, enhanced the beneficial bacteria Lactobacillus and reduced the Pseudomonas, thus maintaining the overall microbiota balance. Taken together, the administration of MR-7 could effectively restore LPS-induced intestine function disorder in turbot by ameliorating inflammatory response, mucosal barrier dysfunction and microbiota dysbiosis.
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http://dx.doi.org/10.1016/j.fsi.2020.04.068DOI Listing
July 2020

Current development of COVID-19 diagnostics, vaccines and therapeutics.

Microbes Infect 2020 Jul - Aug;22(6-7):231-235. Epub 2020 May 6.

Lindsley F. Kimball Research Institute, New York Blood Center, New York, USA; Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Fudan University, Shanghai, China. Electronic address:

A novel coronavirus, designated as SARS-CoV-2, first emerged in Wuhan City, Hubei Province, China, in late December 2019. The rapidly increasing number of cases has caused worldwide panic. In this review, we describe some currently applied diagnostic approaches, as well as therapeutics and vaccines, to prevent, treat and control further outbreaks of SARS-CoV-2 infection.
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http://dx.doi.org/10.1016/j.micinf.2020.05.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7200352PMC
August 2020

Construction of arsenic-metal complexes loaded nanodrugs for solid tumor therapy: A mini review.

Int J Pharm 2020 Jun 4;583:119385. Epub 2020 May 4.

Department of Pharmacy, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China. Electronic address:

Arsenic trioxide (AsO), a front-line therapeutic agent against acute promyelocytic leukemia, has a broad spectrum against malignancies. Unfortunately, the clinical application of AsO in treating hematological cancers has not been transformed to solid tumors, for its dose-limited toxicity and undesirable pharmacokinetics. The ordinary AsO loaded nanodrugs (such as liposomes, polymer micelles, albumin-based nanodrugs, and silica-based nanodrugs, etc.) still could not fuel up pharmaceuticals and eradicate toxicity for low delivery efficiency caused by the instability and severe drug leakage of formulations during circulation. Recently, the approach of forming and delivering arsenic-metal complexes which will dissociate in the tumoral environment caught our mind. This is the most effective strategy to reduce drug leakage in circulation and accumulate arsenite ions in tumor sites, therefore promote the anti-tumor effect and lighten the toxicity of the drug. This review aims to explain the formation mechanism of arsenic-metal nanocomposites and summarize the constructing strategies of the arsenic-metal nanocomplexes (arsenic-nickel, arsenic-manganese, arsenic-platinum, arsenic-gadolinium, arsenic-zinc, and arsenic-iron nanobins) loaded nanodrugs for solid tumor therapy. Furthermore, the expectations and challenges of arsenic-metal complexes containing nanodrugs for cancer therapy in the future were discussed.
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http://dx.doi.org/10.1016/j.ijpharm.2020.119385DOI Listing
June 2020

Effects of dietary raw or Enterococcus faecium fermented soybean meal on growth, antioxidant status, intestinal microbiota, morphology, and inflammatory responses in turbot (Scophthalmus maximus L.).

Fish Shellfish Immunol 2020 May 3;100:261-271. Epub 2020 Mar 3.

Key Laboratory of Aquaculture Nutrition and Feed, Ministry of Agriculture, Ocean University of China, Qingdao, 266003, PR China; Key Laboratory of Mariculture (Ministry of Education), Ocean University of China, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China. Electronic address:

Fermentation has been reported to improve the utilization of plant ingredients including soybean meal (SBM) by fish, but the detailed mechanism is still poorly understood. This study compared the effects of partial replacement of fish meal (FM) protein with SBM or Enterococcus faecium fermented SBM (EFSM) on the growth, antioxidant status, intestinal microbiota, morphology, and inflammatory responses in turbot (Scophthalmus maximus L.). The FM-based diet was used as the control (CONT). Two experimental diets were formulated in which 45% of the FM protein was replaced with SBM or EFSM. Each diet was fed to triplicate groups of fish (7.57 ± 0.01 g) twice daily for 79 d. Inferior growth performance was observed in SBM group, however, no significant depression was observed in EFSM group compared to the CONT group. The CONT group had the highest values of lysozyme, complement component 3, total antioxidant capacity, superoxide dismutase and catalase, followed by the EFSM group, and the lowest in SBM group. The malondialdehyde content was lowest in the CONT group, followed by the EFSM group, and was highest in the SBM group. Gut morphology showed that SBM diet induced alterations typical for intestinal inflammation including decreased villus and microvillus height, and increased width and inflammatory cell infiltration of the lamina propria. However, the EFSM group alleviated such SBM-induced intestinal pathological disruption. Paralleled with the morphological symptoms, the inflammatory gene expression levels of tumor necrosis factor alpha, interleukin-1 beta and interleukin-8 were highest in the SBM group, followed by the EFSM group, and were lowest in the CONT group. Furthermore, the intestinal microbiota analysis revealed that EFSM group had an overall more similar microbiota with CONT group than SBM group. Specifically, compared with the SBM group, EFSM group significantly enhanced the probiotics Lactobacillus and anti-inflammatory bacterium Faecalibaculum, and inhibited the Vibrio. Collectively, this study indicated that Enterococcus faecium fermentation effectively counteracted the negative effects of SBM by enhancing antioxidant capacity, suppressing inflammatory responses, and modulating gut microbiota in turbot.
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http://dx.doi.org/10.1016/j.fsi.2020.02.070DOI Listing
May 2020

DNA Structural Distortions Induced by a Monofunctional Trinuclear Platinum Complex with Various Cross-Links Using Molecular Dynamics Simulation.

J Chem Inf Model 2020 03 4;60(3):1700-1708. Epub 2020 Mar 4.

College of Chemistry, Beijing Normal University, 19# Xinjiekouwai Street, Beijing 100875, China.

The monofunctional trinuclear platinum complex (MTPC), as a promising antitumor agent, can form MTPC-DNA adducts via bifunctional and trifunctional cross-links. Molecular dynamics simulations were used to investigate DNA structural distortions of the MTPC-DNA adducts. MTPC coordinating to DNA results in the decrease of base-pair thermal stability and DNA structural distortions. It is found that there are more significant DNA structural distortions in the trifunctional cross-link than in the bifunctional cross-link, in the 1,4-GG than in the 1,3-GG cross-link, and in the intrastrand than in the interstrand cross-link with the same spans. The results provide a better understanding of DNA structural distortions induced by MTPC with various cross-links at the nucleotide level and are helpful for exploring novel Pt-based anticancer drugs.
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http://dx.doi.org/10.1021/acs.jcim.0c00002DOI Listing
March 2020

An Unmanned Aerial Vehicle-Based Gas Sampling System for Analyzing CO and Atmospheric Particulate Matter in Laboratory.

Sensors (Basel) 2020 Feb 15;20(4). Epub 2020 Feb 15.

College of Mechanical and Electronic Engineering, Northwest A&F University, Yangling, Shaanxi 712100, China.

We developed and tested an unmanned aerial vehicle-based gas sampling system (UGSS) for collecting gases and atmospheric particulate matter (PM). The system applies an alternative way of collecting both vertical and horizontal transects of trace gases in order to analyze them in the laboratory. To identify the best position of the UGSS intake port, aerodynamic flow simulations and experimental verifications of propeller airflow were conducted with an unmanned aerial vehicle (UAV) in hover mode. The UGSS will automatically replace the original gas in the system with gas from a target location to avoid the original gas being stored in the air bags. Experimental results show that the UGSS needs 5 s to replace the system's own original gas using its pump. CO and PM2.5/10 above the corn field are used as the test species to validate the accuracy of the CO gas and PM concentrations collected by UGSS. Deming regression analyses showed good agreement between the measurements from the UGSS and the ground sampling station (y = 1.027x - 11.239, Pearson's correlation coefficient of 0.98 for CO; y = 0.992x + 0.704, Pearson's correlation coefficient of 0.99 for PM).The UGSS provides a measuring method that actively collects gases and PM for manual analyses in the laboratory.
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http://dx.doi.org/10.3390/s20041051DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7070813PMC
February 2020

Activation of Tyrosine Metabolism in CD13+ Cancer Stem Cells Drives Relapse in Hepatocellular Carcinoma.

Cancer Res Treat 2020 Apr 27;52(2):604-621. Epub 2019 Dec 27.

The Third Department of Infectious Diseases, Shanghai Public Health Clinical Center, Shanghai, China.

Purpose: Cancer stem cells (CSCs) are naturally resistant to chemotherapy, explaining why tumor relapse frequently occurs after initial regression upon administration of chemotherapeutic agents in most cases. A CSC population characterized by CD13 expression has been identified in hepatocellular carcinoma (HCC). In the current study, we aimed to clarify the molecular mechanism by which it escapes conventional therapies.

Materials And Methods: Here, we used flow cytometry to examine the percentage of CD13+ CSCs in HepG2 and HuH7 cells after chemotherapy. Using in vitro isotope labeling technique, we compared metabolic pathways between CD13+ and CD13- subpopulations. Using co-immunoprecipitation and western blotting, we determined the target expressions in protein levels under different conditions. We also performed immunohistochemistry to detect the target proteins under different conditions. Animal models were constructed to verify the potential role of tyrosine metabolism in post-chemotherapeutic relapse in vivo.

Results: We observed that quiescent CD13+ CSCs are enriched after chemotherapy in HCCs, and serve as a reservoir for recurrence. Mechanistically, CD13+ CSCs were dependent on aerobic metabolism of tyrosine rather than glucose as energy source. Tyrosine metabolism also generated nuclear acetyl-CoA to acetylate and stabilize Foxd3, thereby allowing CD13+ CSCs cells to sustain quiescence and resistance to chemotherapeutic agents.

Conclusion: These findings encourage further exploration of eliminating CD13+ cells by targeting specific metabolic pathways to prevent recurrence in HCCs.
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http://dx.doi.org/10.4143/crt.2019.444DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7176959PMC
April 2020

AIE nanodots scaffolded by mini-ferritin protein for cellular imaging and photodynamic therapy.

Nanoscale 2020 Jan 14;12(4):2340-2344. Epub 2020 Jan 14.

State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, PR China.

Photodynamic therapy (PDT) is one of the most elegant cancer treatment strategies that can be controlled by a beam of light with non-invasion, precise control, and high spatiotemporal accuracy. An ideal photosensitizer (PS) is the key to ensure the efficacy of PDT. Due to their hydrophobic and rigid planar structures, most traditional PSs are prone to aggregate under physiological conditions, which causes fluorescence quenching and significantly reduces reactive oxygen species (ROS) generation. Fortunately, the emergence of aggregation-induced emission (AIE) dyes offers a potential opportunity to overcome these limitations. When AIE PS molecules are in the aggregation state, the fluorescence intensity and ROS production can be increased. We herein use red AIE PS molecules to prepare stable AIE nanodots for cell imaging and PDT via a simple method with a highly negatively charged mini-ferritin protein as the scaffold. The as-prepared protein-AIE nanodots show strong fluorescence emission and efficient singlet oxygen generation, with good stability, relatively long wavelengths of absorption and emission, and negligible dark toxicity. The mini-ferritin-AIE system may be useful in developing novel functional probes for tumour nanotheranostics.
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http://dx.doi.org/10.1039/c9nr09788kDOI Listing
January 2020

Unmanned Aerial Vehicle-Borne Sensor System for Atmosphere-Particulate-Matter Measurements: Design and Experiments.

Sensors (Basel) 2019 Dec 20;20(1). Epub 2019 Dec 20.

Nanjing Hepu Aviation Technology Co., Ltd., NanJing 211300, China.

An unmanned aerial vehicle (UAV) particulate-matter (PM) monitoring system was developed that can perform three-dimensional stereoscopic observation of PM2.5 and PM10 in the atmosphere. The UAV monitoring system was mainly integrated by modules of data acquisition and processing, wireless data transmission, and global positioning system (GPS). Particularly, in this study, a ground measurement-control subsystem was added that can display and store collected data in real time and set up measurement scenarios, data-storage modes, and system sampling frequency as needed. The UAV PM monitoring system was calibrated via comparison with a national air-quality monitoring station; the data of both systems were highly correlated. Since rotation of the UAV propeller affects measured PM concentration, this study specifically tested this effect by setting up another identical monitoring system fixed at a tower as reference. The UAV systems worked simultaneously to collect data for comparison. A correction method for the propeller disturbance was proposed. Averaged relative errors for the PM2.5 and PM10 concentrations measured by the two systems were 6.2% and 6.6%, respectively, implying that the UAV system could be used for monitoring PM in an atmosphere environment.
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http://dx.doi.org/10.3390/s20010057DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982869PMC
December 2019

Improved utilization of soybean meal through fermentation with commensal Shewanella sp. MR-7 in turbot (Scophthalmus maximus L.).

Microb Cell Fact 2019 Dec 16;18(1):214. Epub 2019 Dec 16.

Key Laboratory of Aquaculture Nutrition and Feed, Ministry of Agriculture, Ocean University of China, 5 Yushan Road, Qingdao, 266003, People's Republic of China.

Background: Increased inclusion of plant proteins in aquafeeds has become a common practice due to the high cost and limited supply of fish meal but generally leads to inferior growth performance and health problems of fish. Effective method is needed to improve the plant proteins utilization and eliminate their negative effects on fish. This study took a unique approach to improve the utilization of soybean meal (SBM) by fish through autochthonous plant-degrading microbe isolation and subsequent fermentation.

Results: A strain of Shewanella sp. MR-7 was isolated and identified as the leading microbe that could utilize SBM in the intestine of turbot. It was further optimized for SBM fermentation and able to improve the protein availability and degrade multiple anti-nutritional factors of SBM. The fishmeal was able to be replaced up to 45% by Shewanella sp. MR-7 fermented SBM compared to only up to 30% by SBM in experimental diets without adverse effects on growth and feed utilization of turbot after feeding trials. Further analyses showed that Shewanella sp. MR-7 fermentation significantly counteracted the SBM-induced adverse effects by increasing digestive enzymes activities, suppressing inflammatory responses, and alleviating microbiota dysbiosis in the intestine of turbot.

Conclusions: This study demonstrated that plant protein utilization by fish could be significantly improved through pre-digestion with isolated plant-degrading host microbes. Further exploitation of autochthonous bacterial activities should be valuable for better performances of plant-based diets in aquaculture.
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http://dx.doi.org/10.1186/s12934-019-1265-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6913000PMC
December 2019

Role of Surface RGD Patterns on Protein Nanocages in Tumor Targeting Revealed Using Precise Discrete Models.

Small 2019 12 25;15(51):e1904838. Epub 2019 Nov 25.

State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, 430071, China.

The effectiveness of active targeting in cancer nanomedicine is becoming increasingly more debatable. Here, the role of the ligand functionalization patterns (number and distribution) on nanoparticle surfaces in tumor targeting is investigated using a 9 nm sized miniferritin protein nanocage, Dps modified with Arg-Gly-Asp (RGD) ligands whose functionalization patterns are precisely controlled. In vitro and in vivo experiments show that RGD modification endows Dps with tumor targeting capacity no matter what the surface pattern is. The tumor targeting of 2-ligand Dps, which is better than that of 1-ligand Dps, rivals or surpasses that of the 12- or 24-ligand Dps. The 12-ligand Dps with clustered RGD distribution shows 2.3 times the in vivo targeting efficiency of that with even distribution. The surface ligand pattern effects are correlated at least to receptor clustering and opsonization. This study provides insights into the understanding of the controversial findings on active tumor targeting in the literature and highlights the necessity of precise functionalization to achieve optimal active targeting in developing cancer nanomedicine.
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http://dx.doi.org/10.1002/smll.201904838DOI Listing
December 2019
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