Publications by authors named "HongLing Ma"

31 Publications

Biochemical, metabolic, and immune responses of mud crab (Scylla paramamosain) after mud crab reovirus infection.

Fish Shellfish Immunol 2022 Jun 30;127:437-445. Epub 2022 Jun 30.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, 510300, PR China. Electronic address:

Mud crab reovirus (MCRV) is a serious pathogen that leads to large economic losses in the mud crab farming. However, the molecular mechanism of the immune response after MCRV infection is unclear. In the present study, physiological, transcriptomic, and metabolomic responses after MCRV infection were investigated. The results showed that MCRV infection could increase lactate dehydrogenase, alanine aminotransferase, and aspartate aminotransferase activities. MCRV infection decreased antioxidant enzyme activity levels, induced oxidative stress, and caused severe histological damage. Transcriptome analysis identified 416 differentially expressed genes, including 354 up-regulated and 62 down-regulated genes. The detoxification, immune response, and metabolic processes-related genes were found. The results showed that two key pathways including phagocytosis and apoptosis played important roles in response to MCRV infection. The combination of transcriptomic and metabolomic analyses showed that related metabolic pathways, such as glycolysis, citrate cycle, lipid, and amino acid metabolism were also significantly disrupted. Moreover, the biosynthesis of unsaturated fatty acids was activated in response to MCRV infection. This study provided a novel insight into the understanding of cellular mechanisms in crustaceans against viral invasion.
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http://dx.doi.org/10.1016/j.fsi.2022.06.058DOI Listing
June 2022

Mechanism of Cadmium Exposure Induced Hepatotoxicity in the Mud Crab (): Activation of Oxidative Stress and Nrf2 Signaling Pathway.

Antioxidants (Basel) 2022 May 17;11(5). Epub 2022 May 17.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China.

Cadmium, one of the most toxic heavy metals, can cause severe oxidative damage to aquatic animals. However, the mechanism whereby the mud crabs respond to cadmium exposure remains unclear. This study investigated the effects of cadmium exposure on oxidative stress and histopathology changes and evaluated the role of the Nrf2 signaling pathway in regulating responses to cadmium-induced hepatotoxicity were investigated in mud crabs. Mud crabs were exposed to 0, 0.01, 0.05, and 0.125 mg/L cadmium for 21 d. The present results indicated that cadmium exposure increased hydrogen peroxide (HO) production, lipid peroxidation and tissue damage, but decreased the activity of superoxide dismutase (SOD) and catalase (CAT), and caused lipid peroxidation and tissue damage. The results of an integrated biomarker index analysis suggested that the toxicity of cadmium was positively related to cadmium concentration. The expression levels of the Nrf2 signaling pathway (Nrf2, metallothionein, and cytochrome P450 enzymes) were up-regulated after cadmium exposure. Silencing of Nrf2 in vivo decreased antioxidant gene (SOD, CAT, and glutathione S-transferase) expression, suggesting that Nrf2 can regulate antioxidant genes. Knocking down Nrf2 in vivo also significantly decreased the activity of SOD and CAT after cadmium exposure. Moreover, silencing of Nrf2 in vivo enhanced HO production and the mortality rates of mud crabs after cadmium exposure. The present study indicated that cadmium exposure induced hepatotoxicity in the mud crab by increasing HO content, which decreased the antioxidant capacity, leading to cell injury. In addition, the Nrf2 is activated to bound with antioxidant response element, initiating the expression of antioxidant enzyme genes during cadmium induced hepatotoxicity in the mud crabs.
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http://dx.doi.org/10.3390/antiox11050978DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9137997PMC
May 2022

Prevalence, virulence genes, and antimicrobial resistance of Vibrio species isolated from diseased marine fish in South China.

Sci Rep 2020 08 31;10(1):14329. Epub 2020 Aug 31.

Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, China.

Here, 70 potential Vibrio pathogens belonging to nine species, dominated by Vibrio harveyi, were isolated and identified from diseased aquacultured marine fish in South China. Subsequently, the prevalence of 11 virulence genes and the resistance to 15 antibiotics in these strains were determined. Most strains possessed atypical virulence genes in addition to typical virulence genes. Notably, hflk and chiA originating from V. harveyi, and flaC associated with V. anguillarum were detected in more than 40% of atypical host strains. Multidrug resistance was widespread: 64.29% strains were resistant to more than three antibiotics, and the multi-antibiotic resistance index ranged from 0.00 to 0.60. The proportions of strains resistant to the antibiotics vancomycin, amoxicillin, midecamycin, and furazolidone all exceeded 50%; nevertheless, all strains were sensitive to florfenicol, norfloxacin, and ciprofloxacin. Furthermore, both virulence genes and antibiotic resistance were more prevalent in Hainan than in Guangdong, owing to the warmer climate and longer annual farming time in Hainan. These results therefore suggest that warming temperatures and overuse of antibiotics are probably enhancing antibiotic resistance and bacterial infection. This study reveals that pathogenic Vibrio spp. with multi-antibiotic resistance are highly prevalent among marine fish in South China and thus warrant further attention. The results will provide helpful guidance for ecological regulation and local antibiotic use in the control of marine fish farming' Vibrio diseases in South China, facilitating the implementation of national green and healthful aquaculture.
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http://dx.doi.org/10.1038/s41598-020-71288-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7459350PMC
August 2020

Phoenixin 14 inhibits ischemia/reperfusion-induced cytotoxicity in microglia.

Arch Biochem Biophys 2020 08 23;689:108411. Epub 2020 May 23.

Department of Neurology, Liaocheng People's Hospital of Shandong First Medical University, Liaocheng City, Shandong Province, 252000, China. Electronic address:

The process of ischemia/reperfusion (IR) in ischemic stroke often leads to significant cell death and permanent neuronal damage. Safe and effective treatments are urgently needed to mitigate the damage caused by IR injury. The naturally occurring pleiotropic peptide phoenixin 14 (PNX-14) has recently come to light as a potential treatment for IR injury. In the present study, we examined the effects of PNX-14 on several key processes involved in ischemic injury, such as pro-inflammatory cytokine expression, oxidative stress, and the related cascade mediated through the toll-like receptor 4 (TLR4) pathway, using BV2 microglia exposed to oxygen-glucose deprivation and reoxygenation (OGD/R). Our results demonstrate an acute ability of PNX-14 to regulate the expression levels of proinflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). PNX-14 also prevented oxidative stress by reducing the generation of reactive oxygen species (ROS) and increasing the level of the antioxidant glutathione (GSH). Importantly, PNX-14 inhibited high-mobility group box 1 (HMGB1)/TLR4/myeloid differentiation primary response 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway, by inhibiting the activation of TLR4 and preventing the nuclear translocation of p65 protein. We further confirmed the cerebroprotective effects of PNX-14 in an MCAO rat model, which resulted in reduced infarct volume and decreased microglia activation. Together, the results of this study implicate a possible protective role of PNX-14 against various aspects of IR injury in vitro.
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http://dx.doi.org/10.1016/j.abb.2020.108411DOI Listing
August 2020

Anti-leucine-rich Glioma Inactivated-1 Encephalitis Associated with Essential Thrombocythemia.

Intern Med 2020 15;59(2):271-275. Epub 2020 Jan 15.

Department of Neurology, Liaocheng People's Hospital; Liaocheng Clinical School, Taishan Medical University, PR China.

Anti-leucine-rich glioma inactivated-1 (anti-LGI1) encephalitis is a subgroup of autoimmune encephalitis. We herein report the case of a 60-year-old man who presented with typical symptoms, including short-term memory loss, mental abnormalities, hyponatremia and seizures characterized by faciobrachial dystonic seizures and who was diagnosed with anti-LGI1 encephalitis. At the same time, he was diagnosed with essential thrombocythemia. A significant improvement was obtained by treatment with corticosteroid, immunoglobulin, mycophenolate mofetil, and hydroxyurea. Autoimmune diseases are associated with a significantly increased risk of developing myeloproliferative neoplasms, which may explain the coexistence of anti-LGI1 encephalitis and essential thrombocythema in this patient; however, but more cases and studies are needed to determine whether there is any correlation between these conditions.
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http://dx.doi.org/10.2169/internalmedicine.2963-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7008042PMC
April 2020

Rapid environmental change shapes pond water microbial community structure and function, affecting mud crab (Scylla paramamosain) survivability.

Appl Microbiol Biotechnol 2020 Mar 8;104(5):2229-2241. Epub 2020 Jan 8.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, China.

The aquatic microbial community is sensitive to environmental change; however, the impacts of those changes combined with disease outbreaks affecting S. paramamosain are unknown. Thus, from March to October, we explored the interaction between aquacultural environmental conditions and microbial composition and function in open-air aquaculture ponds containing S. paramamosain in Southern China. The microbial community structure was significantly positively correlated with microbial community function. The environment variables such as temperature and salinity during May and June changed more quickly compared with other periods, resulting changes in the structure and function of the microbial community affected S. paramamosain survivability, with higher crab mortality observed from May to June compared with other periods. These included changes in the relative abundance of Microtrichales, Synechococcales, Rhodobacterales, Chitinophagales, and SAR11_clade, and corresponding functions associated with glycolysis and/or gluconeogenesis, porphyrin and chlorophyll metabolism, photosynthetic proteins, and transcription factors. These changes could impact S. paramamosain mortality and be used to evaluate the health status of the ponds. Though the environment variables during July~October changed slowly comparing to May and June, the ponds microflora changed which benefit S. paramamosain survivability with correspondingly low S. paramamosain mortality. Therefore, rapid environmental change alters the structure and function of the aquatic microflora, increasing S. paramamosain mortality.
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http://dx.doi.org/10.1007/s00253-019-10328-wDOI Listing
March 2020

Farnesoid X Receptor (FXR) Aggravates Amyloid-β-Triggered Apoptosis by Modulating the cAMP-Response Element-Binding Protein (CREB)/Brain-Derived Neurotrophic Factor (BDNF) Pathway In Vitro.

Med Sci Monit 2019 Dec 8;25:9335-9345. Epub 2019 Dec 8.

Joint Pharmacology Center, Liaocheng People's Hospital, Liaocheng, Shandong, China (mainland).

BACKGROUND Alzheimer's disease (AD), which results in cognitive deficits, usually occurs in older people and is mainly caused by amyloid beta (Aß) deposits and neurofibrillary tangles. The bile acid receptor, farnesoid X receptor (FXR), has been extensively studied in cardiovascular diseases and digestive diseases. However, the role of FXR in AD is not yet understood. The purpose of the present study was to investigate the mechanism of FXR function in AD. MATERIAL AND METHODS Lentivirus infection, flow cytometry, real-time PCR, and western blotting were used to detect the gain or loss of FXR in cell apoptosis induced by Aß. Co-immunoprecipitation was used to analyze the molecular partners involved in Aß-induced apoptosis. RESULTS We found that the mRNA and protein expression of FXR was enhanced in Ab-triggered neuronal apoptosis in differentiated SH-SY5Y cells and in mouse hippocampal neurons. Overexpression of FXR aggravated Aß-triggered neuronal apoptosis in differentiated SH-SY5Y cells, and this effect was further increased by treatment with the FXR agonist 6ECDCA. Molecular mechanism analysis by co-immunoprecipitation and immunoblotting revealed that FXR interacted with the cAMP-response element-binding protein (CREB), leading to decreased CREB and brain-derived neurotrophic factor (BDNF) protein levels. Low expression of FXR mostly reversed the Aß-triggered neuronal apoptosis effect and prevented the reduction in CREB and BDNF. CONCLUSIONS These data suggest that FXR regulates Aß-induced neuronal apoptosis, which may be dependent on the CREB/BDNF signaling pathway in vitro.
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http://dx.doi.org/10.12659/MSM.920065DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6918812PMC
December 2019

Molecular typing of Streptococcus agalactiae isolates of serotype Ia from tilapia in southern China.

FEMS Microbiol Lett 2019 07;366(13)

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, Guangdong Province, PR China.

Streptococcus agalactiae is an important pathogen of tilapia causing enormous economic losses worldwide. In this study, multilocus sequence typing indicated that 75 S. agalactiae isolates from tilapia in southern China belonged to sequence type-7, as well as belonging to serotype Ia, as confirmed by multiplex PCR assay. The putative-virulence gene profiles and genetic variation of these strains were determined by three sets of multiplex PCR and multi-virulence locus sequencing typing (MVLST), respectively. Analysis of putative-virulence gene profiles showed that each strain harbored 18 putative-virulence genes but lacked lmb and scpB. Three putative-virulence genes (srr-1, bibA and fbsA) were further selected for MVLST analysis. Our data showed that the strains had 14 MVLST types (1-14) and clustered in three groups (Groups I-Ⅲ). The period of time during 2013 and 2014 was an important turning point for the differentiation of the putative-virulence genes of S. agalactiae, as type 1 within Group Ⅱ became the predominant MVLST type. There were significant differences in MVLST types of S. agalactiae isolated from different tilapia farming regions. MVLST assay may improve the discriminatory power and is suitable for understanding the epidemiology of S. agalactiae serotype Ia and screening multivalent vaccine candidate strains.
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http://dx.doi.org/10.1093/femsle/fnz154DOI Listing
July 2019

Expression of lactate dehydrogenase is induced during hypoxia via HIF-1 in the mud crab Scylla paramamosain.

Comp Biochem Physiol C Toxicol Pharmacol 2019 Nov 2;225:108563. Epub 2019 Jul 2.

Key Laboratory of Aquatic Product Processing, Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong 510300, PR China; Shanghai Ocean University, Shanghai 201206, PR China. Electronic address:

Lactate dehydrogenase (LDH) is a key enzyme involved in anaerobic metabolism in most organisms. In the present study, we determined the structure and function of LDH sequence in Scylla paramamosain (SpLDH) by gene cloning, expression and RNA interference techniques in order to explore the genetic characteristics of LDH and its relationship with HIF-1 during hypoxia. The full-length cDNA was 1453 bp with an open reading frame (ORF) of 996 bp, and encoded a polypeptide of 332 amino acids. Homology analysis showed that the SpLDH gene is highly similar to arthropods. The SpLDH transcript increased after hypoxia in all tested tissues. The silencing of HIF-1 blocked the increase in LDH mRNA and activity, which were induced by hypoxia in gill and muscle tissues. Our results indicated that SpLDH expression was regulated transcriptionally by HIF-1.
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http://dx.doi.org/10.1016/j.cbpc.2019.108563DOI Listing
November 2019

Genetic basis of surfactant dysfunction in Chinese children: A retrospective study.

Pediatr Pulmonol 2019 08 12;54(8):1173-1181. Epub 2019 May 12.

National Clinical Research Center for Respiratory Diseases, Department of Respiratory Medicine, Beijing Children's Hospital, Capital Medical University, and the National Center for Children's Health, Beijing, China.

Objective: To investigate the prevalence of surfactant dysfunction (SD) and the genotype distribution in Chinese childhood interstitial lung disease (chILD).

Methods: From December 2013 to December 2016, whole exons and splicing regions of surfactant protein (SP)-B, SP-C, and adenosine triphosphate (ATP)-binding cassette subfamily A member 3 (ABCA3) were sequenced in chILD with unknown etiology in five children's medical centers of China. The sequencing was performed by Next-generation sequencing technique in a molecular genetics laboratory. The clinical and genetic data were reviewed retrospectively.

Results: In total, 136 patients of age 3 months to 13 years (mean 12.5 ± 9.4 months) were recruited, among which 76 were males. Of the 136 cases of chILD, 13.2% (18 of 136) were diagnosed with SD. In these 18 SD cases, 15 had heterozygous SP-C deficiencies, two cases had compound heterozygous ABCA3 deficiencies, and no SP-B deficiency was identified. In SP-C deficiencies, there were six cases with p.I73T, 2 with p.I73N, 5 with p.V39L, 1 with c.417delA, and 1 case with IVS4, +1G>C. Two cases of ABCA3 mutation were heterozygous with c.1755delC and c.2890G>A; c.3913T>C (R1305W) and exon 13 to 18 deletion. One was negative by sequencing while diagnosed positive by pathology.

Conclusion: The proportion of genetic mutation of SD in chILD is 13.2% in China, of which SP-C deficiency is predominant. The mutation, SP-C p.V39L, was found to be relatively prevalent in China and warrants further investigation.
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http://dx.doi.org/10.1002/ppul.24334DOI Listing
August 2019

Corrigendum: Identification of a Novel Small RNA in ZJ-T and Its Characterization With Phenotype MicroArray Technology.

Front Microbiol 2019 31;10:21. Epub 2019 Jan 31.

Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, China.

[This corrects the article DOI: 10.3389/fmicb.2018.02394.].
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http://dx.doi.org/10.3389/fmicb.2019.00021DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6365953PMC
January 2019

Coupled changes of bacterial community and function in the gut of mud crab (Scylla Paramamosain) in response to Baimang disease.

AMB Express 2019 Feb 2;9(1):18. Epub 2019 Feb 2.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 309 of Building Keyan, 231 Xingang Xi Road, Guangzhou, China.

Increasing evidence has revealed a close association between intestinal bacterial community and hosts health. However, it is unclear whether and what extend Baimang disease alters the intestinal microbiota in mud crab (Scylla paramamosain). Here, we conducted intestinal contents Illumina sequencing of healthy and Baimang diseased mud crab (S. paramamosain) to understand bacterial community variations among health status. In addition, bacterial functional predication was used to investigate whether and how the bacteria variations further change their functions? The phyla of Proteobacteria, Fusobacteria, Cyanobacteria, Tenericutes, Firmicutes, Bacteroidetes, and Spirochaetae constituted over 96.44% of the total intestinal bacteria, with being the dominant taxa. The 7 most significantly different orders, including the increased four orders of Clostridiales, Entomoplasmatales, Bacteroidales, and Mycoplasmatales and the decreased three orders of Vibrionales, Campylobacterales, and Fusobacteriales, accounted for 61.14% dissimilarity, probably being the indicator taxa of Baimang disease. Accordingly, 12 Kyoto Encyclopedia of Genes and Genomes orthologies in level 3 shifted significantly at the diseased crabs. Especially, bacterial secretion system, secretion system, lipopolysaccharide biosynthesis proteins and Vibrio cholerae pathogenic cycle, being related to bacterial virulence, were reduced. In addition, the reduced butanoate metabolism, and induced methane metabolism and one carbon pool by folate were important metabolic processes of probiotic, such as Bacteroides spp. and Clostridium spp., with playing critical roles in host health. This study suggests that Baimang disease coupled altered the intestinal bacterial communities and functions, providing timely information for further analysis the influencing mechanism of Baimang disease in mud crab (S. paramamosain).
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http://dx.doi.org/10.1186/s13568-019-0745-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6359999PMC
February 2019

Spatial and temporal variation of antibiotic resistance in marine fish cage-culture area of Guangdong, China.

Environ Pollut 2019 Mar 12;246:463-471. Epub 2018 Dec 12.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, China. Electronic address:

The rapid emergence and dissemination of antibiotic resistance poses a threat to human health and to the marine environment. We have investigated the abundance and diversity of antibiotic resistance genes (ARGs) and of antibiotic-resistant bacteria (ARB), during the seedling period, rearing period, and harvesting period in seven marine fish cage-culture areas in Guangdong. Spatial and temporal variations of AGRs and ARB were also analyzed. Culture-based methods and quantitative PCR were used to detect ARB and ARGs. Bacterial resistance rates were no significantly different within farming periods. The proportion of antibiotic-resistant bacteria was extremely low (average on 1.15%), except for oxytetracycline-resistant bacteria (average on 34.15%). Vibrio was the most common ARB. Sul1, tetB, and ermB, had the highest relative abundance. The abundance of ARGs in the harvesting period was significant highest. The total abundance of ARGs was highest at Raoping and lowest at Dayawan and Liusha. Most ARGs were associated with opportunistic pathogens. The environmental factors effecting ARB and ARGs are complex, and no key factors were identified. This study provides a theoretical basis for assessing the harmfulness of ARGs and ARB to food safety and human health.
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http://dx.doi.org/10.1016/j.envpol.2018.12.024DOI Listing
March 2019

Identification of a Novel Small RNA in ZJ-T and Its Characterization With Phenotype MicroArray Technology.

Front Microbiol 2018 5;9:2394. Epub 2018 Oct 5.

Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, China.

Small non-coding RNAs (sRNAs) are important modulators of gene expression and are involved in the pathogenesis and survival of prokaryotes. However, few studies have been conducted with , which limits our ability to probe the regulation of virulence and environmental adaptation by sRNAs in this opportunistic pathogen. In this study, the sRNA candidate was identified in ZJ-T. The precise transcript end, secondary structure, and sequence conservation were determined. A null mutant was constructed and characterized by using Phenotype MicroArray (PM) technology. target prediction was conducted by IntaRNA and TargetRNA2. Subsequently, a 107 nt transcript was validated with a sigma70 promoter at the 5' end and a Rho-independent terminator at the 3' end. The sRNA had four stem-loop structures and was conserved among , and . Deletion of in ZJ-T led to a weaker utilization of D-mannose, D-melibiose, lactulose, and inosine as carbon sources but stronger utilization of L-cysteine as nitrogen source. Moreover, the mutant showed stronger resistance to osmotic stress but weaker resistance to pH stress. Additionally, a total of 22 common targets were identified and several were related to the observed phenotype of the mutant. This study indicated that the novel sRNA, , is conserved and restricted to spp., affecting the utilization of several carbon and nitrogen sources and the response to osmotic and pH stress. These results extend our understanding of sRNA regulation in and provide a significant resource for the further study of the precise target mRNAs of , which may provide targets for antibacterial therapeutic or attenuated vaccines against spp.
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http://dx.doi.org/10.3389/fmicb.2018.02394DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6186989PMC
October 2018

Mathematical model of salt cavern leaching for gas storage in high-insoluble salt formations.

Sci Rep 2018 01 10;8(1):372. Epub 2018 Jan 10.

State Key Laboratory of Geomechanics and Geotechnical Engineering, Institute of Rock and Soil Mechanics, Chinese Academy of Sciences, Wuhan, 430071, Hubei, China.

A mathematical model is established to predict the salt cavern development during leaching in high-insoluble salt formations. The salt-brine mass transfer rate is introduced, and the effects of the insoluble sediments on the development of the cavern are included. Considering the salt mass conservation in the cavern, the couple equations of the cavern shape, brine concentration and brine velocity are derived. According to the falling and accumulating rules of the insoluble particles, the governing equations of the insoluble sediments are deduced. A computer program using VC++ language is developed to obtain the numerical solution of these equations. To verify the proposed model, the leaching processes of two salt caverns of Jintan underground gas storage are simulated by the program, using the actual geological and technological parameters. The same simulation is performed by the current mainstream leaching software in China. The simulation results of the two programs are compared with the available field data. It shows that the proposed software is more accurate on the shape prediction of the cavern bottom and roof, which demonstrates the reliability and applicability of the model.
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http://dx.doi.org/10.1038/s41598-017-18546-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5762648PMC
January 2018

ORP4L Facilitates Macrophage Survival via G-Protein-Coupled Signaling: ORP4L-/- Mice Display a Reduction of Atherosclerosis.

Circ Res 2016 Dec 11;119(12):1296-1312. Epub 2016 Oct 11.

From the Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Department of Biotechnology, College of Life Science and Technology, Jinan University, Guangzhou, China (W.Z., G.P., L.W., J.L., B.Z., X.C., H.M., C.L., D.Y.); Minerva Foundation Institute for Medical Research, Biomedicum 2U, Helsinki, Finland (S.L., V.M.O.); Division of Cardiac Surgery, the First Affiliated Hospital (J.O.) and Research Center for Drug Discovery, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China (M.X., J.X.); and U1011 Inserm, EGID, Université Lille, CHU Lille, Institut Pasteur de Lille, France (B.S.).

Rationale: Macrophage survival within the arterial wall is a central factor contributing to atherogenesis. Oxysterols, major components of oxidized low-density lipoprotein, exert cytotoxic effects on macrophages.

Objective: To determine whether oxysterol-binding protein-related protein 4 L (ORP4L), an oxysterol-binding protein, affects macrophage survival and the pathogenesis of atherosclerosis.

Methods And Results: By hiring cell biological approaches and ORP4L mice, we show that ORP4L coexpresses with and forms a complex with Gα and phospholipase C (PLC)-β3 in macrophages. ORP4L facilitates G-protein-coupled ligand-induced PLCβ3 activation, IP production, and Ca release from the endoplasmic reticulum. Through this mechanism, ORP4L sustains antiapoptotic Bcl-XL expression through Ca-mediated c-AMP responsive element binding protein transcriptional regulation and thus protects macrophages from apoptosis. Excessive stimulation with the oxysterol 25-hydroxycholesterol disassembles the ORP4L/Gα/PLCβ3 complexes, resulting in reduced PLCβ3 activity, IP production, and Ca release, as well as decreased Bcl-XL expression and increased apoptosis. Overexpression of ORP4L counteracts these oxysterol-induced defects. Mice lacking ORP4L exhibit increased apoptosis of macrophages in atherosclerotic lesions and a reduced lesion size.

Conclusions: ORP4L is crucial for macrophage survival. It counteracts the cytotoxicity of oxysterols/oxidized low-density lipoprotein to protect macrophage from apoptosis, thus playing an important role in the development of atherosclerosis.
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http://dx.doi.org/10.1161/CIRCRESAHA.116.309603DOI Listing
December 2016

[The significance of the monocyte human leukocyte antigen-DR level in the assessment of the severity of acute pancreatitis].

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 2016 Mar;28(3):221-4

Objective: To investigate the clinical value of the peripheral blood monocyte human leukocyte antigen-DR (mHLA-DR) for assessment of degree of severity and the diagnosis of acute pancreatitis (AP).

Methods: A case-control study was conducted. Eighty-six AP patients admitted to Shandong Liaocheng People's Hospital from June 2014 to May 2015 were enrolled. Patients were classified into four groups [mild (n = 33), moderate (n = 25), severe (n = 16), critical (n = 12)] according to the disease classification. Eighty healthy persons subjected to physical examination center of our hospital at the same time were served as controls. Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores in patients were estimated. Flow cytometry was used to measure the expression of the peripheral blood mHLA-DR, and the Pearson method was used to analyze the relationship between the level of mHLA-DR and the APACHE Ⅱ score. The receiver-operating characteristic curve (ROC) was plotted, and then the clinical value of the peripheral blood mHLA-DR was analyzed for the diagnostic value in AP patients.

Results: The expression of the mHLA-DR in patients with AP was significantly lower than that of healthy control group [(63.7±18.6)% vs. (86.4±8.3)%, t = 5.319, P < 0.001]. The expression levels of the mHLA-DR in mild group, moderate group, severe group, and critical group were (79.6±6.5)%, (66.4±9.4)%, (49.9±8.1)%, (32.5±12.0)%, respectively, and the APACHE Ⅱ score were 4.67±1.99, 5.88±2.05, 9.06±2.62, 12.33±3.96, respectively. Pair wise comparisons were statistically significant (all P < 0.05). The HLA-DR expression level in the peripheral blood of patients with AP was negatively correlated with the APACHE Ⅱ score (r = -0.695, P < 0.001). The area under the ROC curve (AUC) of mHLA-DR expression in peripheral blood for AP was 0.894 [95% confidence interval (95%CI) = 0.847-0.941, P < 0.001], and the cut-off point was 84.40%, with the sensitivity of 75.0%, the specificity of 90.7%, and the accuracy rate of 83.1%. The AUC of mHLA-DR expression for mild AP was 0.938 (95%CI = 0.889-0.987, P < 0.001), and the cut-off point was 72.70%, with the sensitivity of 87.9%, the specificity of 88.7%, and the accuracy rate of 88.4%. The AUC of mHLA-DR expression for severe and critical AP was 0.943 (95%CI = 0.881-1.005, P < 0.001), and the cut-off point was 57.85%, with the sensitivity of 84.0%, the specificity of 96.4%, and the accuracy rate of 90.6%.

Conclusions: The expression levels of the peripheral blood mHLA-DR in AP patients can reflect the degree of disease, and contribute to the diagnosis of AP. The value of mHLA-DR may be used as a new biological indicator in the diagnosis and assessment for the severity of AP.%
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March 2016

[Trisomy 21 syndrome associated interstitial lung disease: a case report].

Zhonghua Er Ke Za Zhi 2015 Oct;53(10):771-4

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Objective: To study the pathology, imaging and clinical features of a child with trisomy 21 syndrome associated interstitial lung disease.

Method: Data of a case with trisomy 21 syndrome associated interstitial lung disease confirmed by lung imaging and pathology were collected, analyzed and the related reports in literature were reviewed.

Result: The patient was a one year and 7 months old boy who suffered from severe pneumonia and recurrent infection during his hospital stay. When his disease was stable, he did not have shortness of breath and cyanosis, but a chest computed tomography (CT) showed ground-glass opacity, regional emphysema, band-like change in lung parenchyma, which indicated interstitial lung diseases. Unequal air inflation in bilateral lungs and diffuse over-distension of peripheral air spaces in lung surface were seen through thoracoscope. Pathological examination indicated that alveolar, alveolar ducts and alveolar sac were enlarged, alveolar septa was expanded. There were two reports in lung pathology of trisomy 21 syndrome, alveolar growth abnormalities was seen in 86%-88% cases. The multiple subpleural cysts in chest CT was characteristic. Clinically, trisomy 21 syndrome had high morbidity of respiratory tract infection and progress to respiratory failure frequently. Prolonged postoperative desaturation was constant which required long duration of respiratory support.

Conclusion: Trisomy 21 syndrome associated alveolar growth abnormalities were confirmed, which manifest as alveolar simplification in pathology and interstitial lung diseases in imaging. The risk of respiratory failure in these cases caused by infection and surgery should be considered.
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October 2015

The complete genome of a viable archaeum isolated from 123-million-year-old rock salt.

Environ Microbiol 2016 Feb 18;18(2):565-79. Epub 2016 Jan 18.

Department of Biosciences, Institute of Biotechnology, University of Helsinki, Helsinki, Finland.

Live microbes have been isolated from rock salt up to Permian age. Only obligatory cellular functions can be performed in halite-buried cells. Consequently, their genomic sequences are likely to remain virtually unchanged. However, the available sequence information from these organisms is scarce and consists of mainly ribosomal 16S sequences. Here, live archaea were isolated from early Cretaceous (∼ 123 million years old) halite from the depth of 2000 m in Qianjiang Depression, Hubei Province, China. The sample was radiologically dated and subjected to rigorous surface sterilization before microbe isolation. The isolates represented a single novel species of Halobacterium, for which we suggest the name Halobacterium hubeiense, type strain Hbt. hubeiense JI20-1. The species was closely related to a Permian (225-280 million years old) isolate, Halobacterium noricense, originating from Alpine rock salt. This study is the first one to publish the complete genome of an organism originating from surface-sterilized ancient halite. In the future, genomic data from halite-buried microbes can become a key factor in understanding the mechanisms by which these organisms are able to survive in harsh conditions deep underground or possibly on other celestial bodies.
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http://dx.doi.org/10.1111/1462-2920.13130DOI Listing
February 2016

Epidemiological characterization of VNNV in hatchery-reared and wild marine fish on Hainan Island, China, and experimental infection of golden pompano (Trachinotus ovatus) juveniles.

Arch Virol 2015 Dec 9;160(12):2979-89. Epub 2015 Sep 9.

Key Laboratory of Aquatic Product Processing, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, Guangdong, People's Republic of China.

The current epidemiological situation of viral nervous necrosis virus (VNNV) on Hainan Island was investigated. A total of 490 hatchery-reared fish and 652 wild fish were sampled for VNNV detection from March 2013 to May 2014. Positive detection rates of 84.53% (153/181) and 0.97 % (3/309) were obtained in diseased and healthy hatchery-reared samples, respectively, by conventional RT-PCR. However, using more-sensitive nested RT-PCR, the positive detection rates in healthy hatchery-reared fish reached up to 64.08% (198/309), suggesting that asymptomatic VNNV carriers commonly exist among larvae and juveniles breeding on Hainan Island. In wild-fish samples, 2.6% (17/652) and 34.2% (223/652) positive detection rates were observed using RT-PCR and nested RT-PCR, respectively, indicating that wild fish may be a potential reservoir for VNNV. Phylogenetic analysis showed that all 52 VNNV isolates from cultured fish belong to the RGNNV genotype, but 2 out of 48 VNNV isolates from wild fish samples were found to be of the SJNNV genotype. This study is the first to confirm the existence of SJNNV-genotype VNNV in China. Golden pompano, an important fish species for culture, was selected as a fish model to investigate the optimal conditions for RGNNV disease progression in artificial infection experiments. The effects of temperature, salinity, and fish size were evaluated. Results showed that 28 °C and 20 ‰ are the optimal infection temperature and salinity, respectively, and golden pompano juveniles with small body sizes are more susceptible to RGNNV. These findings are highly consistent with those conditions involved in the natural outbreak of RGNNV.
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http://dx.doi.org/10.1007/s00705-015-2590-0DOI Listing
December 2015

Susceptibility of farmed juvenile giant grouper Epinephelus lanceolatus to a newly isolated grouper iridovirus (genus Ranavirus).

Vet Microbiol 2015 Jun 24;177(3-4):270-9. Epub 2015 Mar 24.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, The South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, PR China. Electronic address:

A ranavirus was isolated from the diseased farmed groupers (Grouper iridovirus in genus Ranavirus, GIV-R), Epinephelus hybrids (blotchy rock cod, Epinephelus fuscoguttatus ♀×giant grouper, Epinephelus lanceolatus ♂), in Sanya, Hainan, in July 2013. In this study, susceptibility of farmed juvenile giant grouper E. lanceolatus to GIV-R was determined by intraperitoneally injection. The cumulative mortality reached to 81% at 5 day post infection. Histologically, severe degeneration with massive pycnotic nuclei in spleen and kidney tissues was observed, and some small-size inclusion body-bearing cells (IBCs) existed in spleen. Hemorrhage and infiltration of inflammatory cells were presented in gill, liver and heart along with tissue degeneration and necrosis of varying severity. The results of immunohistochemistry analysis showed that the strongest immunolabellings were obtained from the kidney and spleen tissues, while intermediate intensity signals were observed in the heart, stomach, gill and liver tissues, and the weakest signals were obtained from the intestine and brain, but no signal was obtained in eyes. Electron microscopy revealed that spleen of moribund fish contained many viral particles in cytoplasm. Interestingly, in surviving fish, abnormal hypertrophic cells were observed in both splenic corpuscle and renal corpuscle, while no hypertrophic cell was observed in the other parts of spleen and kidney tissues. Moreover, immunolabellings only stained the hypertrophic cells in splenic corpuscle and renal corpuscle. This indicated that splenic corpuscle and renal corpuscle play an important role in GIV-R infection and replication.
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http://dx.doi.org/10.1016/j.vetmic.2015.03.017DOI Listing
June 2015

[Analysis of 3 cases with Mycoplasma pneumoniae-associated hemophagocytic syndrome and review of literature].

Zhonghua Er Ke Za Zhi 2014 Oct;52(10):792-6

Department of Respiratory Disease, Shenzhen Children's Hospital, Shenzhen 518026, China. Email:

Objective: To analyze the clinical characteristics of Mycoplasma pneumoniae-associated hemophagocytic syndrome (MP-HLH).

Method: A retrospective investigation of the clinical manifestation, laboratory test, imagelogy, clinical course and outcome of 3 cases with MP-HLH seen between June 2013 and July 2013 in Shenzhen Children's Hospital, and review of relevant literature were conducted.

Result: Of the 3 cases of MP-HLH, 2 were males, one was female, the ages were 1 year, 3 years and 6 years, respectively. They had no underlying disease previously. All the 3 cases had onset of fever, cough as main symptoms. Diagnosis of refractory Mycoplasma pneumoniae pneumonia was made, which was accompanied by decreased neutrophils [(0.08-0.68)×10(9)/L], hemoglobin [(79-103) g/L], platelet [(64-157)×10(9)/L], plasma fibrinogen [(1.3-1.5) g/L], lactate dehydrogenase [(1,170-1,285) U/L] and increased serum ferritin [(936.7-39 789.0) µg/L] in the third week of course. In two cases the T lymphocytes decreased, and the NK cell activity decreased significantly in one. Bone marrow cytology showed prompted bone marrow hyperplasia, and the phenomenon of phagocytosed blood cells. CT scan was performed for all the cases and consolidation with pleural effusion were shown. Two cases were admitted to PICU, and required endotracheal intubation and mechanical ventilation. Flexible bronchoscopy and bronchial lavage were performed and bronchial cast was found in two cases. All of them were treated with macrolide combined with other antibiotics, glucocorticoids and gamma globulin combination therapy, including one case given dexamethasone [10 mg/(m2·d)], cyclosporine[6 mg/(kg·d)], etoposide [150 mg/(m2·d)] chemotherapy. Two cases were cured, and 1 case died. The authors summarized the 18 cases reported in domestic and foreign literature. Foreign children were diagnosed and treated with steroids in 1-2 weeks, and 10 cases were cured, and 2 cases died. They died of massive hemorrhage and meningoencephalitis, and domestic children were diagnosed and treated within two to 4 weeks after onset, 5 cases were cured, one case died of severe pneumonia.

Conclusion: MP-HLH is a rare disease in children, and had acute onset, rapid progression and high mortality. Early treatment with steroids was associated with a good prognosis, the key to successful treatment is early diagnosis and treatment, avoiding the immune cascade. Too late a diagnosis or development of serious complications may lead to death.
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October 2014

Halophilic archaea cultivated from surface sterilized middle-late eocene rock salt are polyploid.

PLoS One 2014 22;9(10):e110533. Epub 2014 Oct 22.

Institute of Biotechnology and Department of Biosciences, University of Helsinki, Helsinki, Finland.

Live bacteria and archaea have been isolated from several rock salt deposits of up to hundreds of millions of years of age from all around the world. A key factor affecting their longevity is the ability to keep their genomic DNA intact, for which efficient repair mechanisms are needed. Polyploid microbes are known to have an increased resistance towards mutations and DNA damage, and it has been suggested that microbes from deeply buried rock salt would carry several copies of their genomes. Here, cultivable halophilic microbes were isolated from a surface sterilized middle-late Eocene (38-41 million years ago) rock salt sample, drilled from the depth of 800 m at Yunying salt mine, China. Eight unique isolates were obtained, which represented two haloarchaeal genera, Halobacterium and Halolamina. We used real-time PCR to show that our isolates are polyploid, with genome copy numbers of 11-14 genomes per cell in exponential growth phase. The ploidy level was slightly downregulated in stationary growth phase, but the cells still had an average genome copy number of 6-8. The polyploidy of halophilic archaea living in ancient rock salt might be a factor explaining how these organisms are able to overcome the challenge of prolonged survival during their entombment.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0110533PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4206341PMC
June 2015

Elevation in circulating YKL-40 concentration in patients with cerebrovascular disease.

Bosn J Basic Med Sci 2014 Aug 14;14(3):120-4. Epub 2014 Aug 14.

Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou.

YKL-40 is a novel inflammatory protein. Elevated serum levels of YKL-40 have been reported in patients with atherosclerosis and other cardiovascular diseases, but the circulating profile of YKL-40 in patients with cerebrovascular disease has been less investigated. This prospective observational study aimed to determine serum levels of YKL-40 in patients with different subtypes and severities of cerebrovascular disease. Eighty patients with acute ischemic stroke, 30 patients with acute hemorrhagic stroke, 15 patients with transient ischemic attack (TIA) and 18 age- and gender-matched healthy control subjects were recruited. Blood was sampled. Serum levels of YKL-40 were measured by ELISA. In healthy control subjects, serum levels of YKL-40 were 45.09 ± 31.41 ng/ml, significantly lower than those in patients with acute ischemic stroke (178.58 ± 127.78 ng/ml), hemorrhagic stroke (105.32 ± 87.35 ng/ml) and TIA (148.09 ± 108 ng/ml) respectively (P<0.05). When the 80 acute ischemic stroke cases were stratified into four Oxfordshire Community Stroke subtypes, serum levels of YKL-40 were significantly higher in patients with total anterior (n=16), partial anterior (n=25) and posterior (n=12) circulation infarctions respectively than those with lacunar (n=27) infarction (P<0.05). Moreover, 63 of 80 patients with acute ischemic stroke survived. Circulating levels of YKL-40 in these stroke survivors were associated with the United States National Institutes of Health Stroke Scale (NIHSS) scores of neurological deficit. In summary, serum levels of YKL-40 were elevated in patients with cerebrovascular disease in lesion subtype- and severity-dependent manners. These observations suggest a potential for YKL-40 as a diagnostic/prognostic biomarker for cerebrovascular disease.
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http://dx.doi.org/10.17305/bjbms.2014.3.42DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4333993PMC
August 2014

Rapid detection of mud crab dicistrovirus-1 using loop-mediated isothermal amplification.

J Virol Methods 2014 Nov 27;208:171-6. Epub 2014 Aug 27.

Key Laboratory of Aquatic Product Processing, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China.

Mud crab dicistrovirus-1 (MCDV-1) was isolated from the mud crab (Scylla paramamosain), resulting in mass mortality and widespread economic loss in China. In this study, a detection method for MCDV-1 using loop-mediated isothermal amplification was developed. Two pairs of primers targeting the VP2 gene were designed. These primers were the outer primers F3 and B3, and the inner primers FIP and BIP. Optimal amplification was carried out using 0.2 μmol/L F3/B3, 1.6 μmol/L FIP/BIP, 6 mmol/L Mg(2+), 0.8 mmol/L dNTPs, and 0.8 mol/L betaine, and completed in 1h at 62°C. The products demonstrated a ladder pattern on agarose gel electrophoresis and could also be detected visually according to turbidity, or by adding SYBR Green I and observing a color change from orange to green. The proposed method could specifically amplify MCDV-1 gene fragments. Sensitivity assay revealed that six copies of the viral genome could be detected by this method, which was 1000-fold more sensitive than that of conventional PCR using constructed plasmid as amplification template. At clinical sample level, sensitivity of LAMP was 100-fold higher than that of conventional PCR.
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http://dx.doi.org/10.1016/j.jviromet.2014.08.014DOI Listing
November 2014

[Clinical analysis of 4 children with negative pressure pulmonary edema].

Zhonghua Er Ke Za Zhi 2014 Feb;52(2):122-7

Objective: To analyze the clinical characteristics of negative pressure pulmonary edema (NPPE).

Method: A retrospective investigation of the clinical manifestation, imageology, clinical course and outcome of 4 children with NPPE seen between June 2012 and July 2013 in a children's hospital. The causation of the airway obstruction was also explored.

Result: All the 4 cases were boys, the range of age was 40 days to 9 years. They had no history of respiratory and circulatory system disease. In 3 cases the disease had a sudden onset after the obstruction of airway, and in one the onset occurred 1.5 hours after removing the airway foreign body. All these cases presented with tachypnea, dyspnea, and cyanosis, none had fever. Three cases had coarse rales. Chest radiography was performed in 3 cases and CT scan was performed in 1 case, in all of them both lungs displayed diffuse ground-glass-like change and patchy consolidative infiltrates. Three cases were admitted to the ICU, duration of mechanical ventilation was less than 24 hours in 2 cases and 39 hours in one. Oxygen was given by mask to the remaining one in emergency department, whose symptoms were obviously improved in 10 hours. None was treated with diuretics, glucocorticoids or inotropic agents. Chest radiographs were taken within 24 hours of treatment in 2 cases and 24-48 hours in the other 2; almost all the pulmonary infiltrates were resolved. All the 4 cases were cured. The causes of airway obstruction were airway foreign bodies in two cases, laryngospasm in one and laryngomalacia in the other.

Conclusion: NPPE is a life-threatening emergency, which is manifested by rapid onset of respiratory distress rapidly (usually in several minutes, but might be hours later) after relief of the airway obstruction, with findings of pulmonary edema in chest radiograph. The symptoms resolve rapidly by oxygen therapy timely with or without mechanical ventilation. In children with airway obstruction, NPPE should be considered.
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February 2014

Increased atherosclerotic lesions in LDL receptor deficient mice with hematopoietic nuclear receptor Rev-erbα knock- down.

J Am Heart Assoc 2013 Aug 20;2(4):e000235. Epub 2013 Aug 20.

Department of Biotechnology, Jinan University, Guangzhou, China.

Background: Nuclear receptor Rev-erbα plays important roles in circadian clock timing, lipid metabolism, adipogenesis, and vascular inflammation. However, the role of Rev-erbα in atherosclerotic lesion development has not been assessed in vivo.

Methods And Results: The nuclear receptor Rev-erbα was knocked down in mouse haematopoietic cells by means of shRNA-lentiviral transduction, followed by bone marrow transplantation into LDL receptor knockout mice. The Rev-erbα protein in peripheral macrophage was reduced by 70% as compared to control mice injected with nontargeting shRNA lentivirus-transduced bone marrow. A significant increase in atherosclerotic lesions was observed around the aorta valves as well as upon en face aorta analysis of Rev-erbα knock-down bone marrow recipients (P<0.01) as compared to the control mice, while plasma cholesterol, phospholipid, and triacylglycerol levels were not affected. Overexpression of Rev-erbα in bone marrow mononuclear cells decreased inflammatory M1 while increasing M2 macrophage markers, while Rev-erbα knock down increased the macrophage inflammatory phenotype in vitro and in vivo. Furthermore, treatment of differentiating macrophages with the Rev-erbα ligand heme promoted expression of antiinflammatory M2 markers.

Conclusions: These observations identify hematopoietic cell Rev-erbα as a new modulator of atherogenesis in mice.
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http://dx.doi.org/10.1161/JAHA.113.000235DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3828791PMC
August 2013

OSBP-related protein 7 interacts with GATE-16 and negatively regulates GS28 protein stability.

Exp Cell Res 2011 Oct 6;317(16):2353-63. Epub 2011 Jun 6.

Department of Biotechnology, Jinan University, Guangzhou, China.

ORP7 is a member of oxysterol-binding protein (OSBP) family, the function of which has remained obscure. In this study, we identified by yeast two-hybrid screening an interaction partner of ORP7, GATE-16, which (i) regulates Golgi SNARE of 28kDa (GS28) function and stability, and (ii) plays a role in autophagosome biogenesis. The interaction was confirmed by bimolecular fluorescence complementation (BiFC) assay in living cells. The interacting regions were delineated within aa 1-142 of ORP7 and aa 30-117 of GATE-16. ORP7 knock-down in 293A cells resulted in a 40% increase of GS28 protein while ORP7 overexpression had the opposite effect (25% decrease of GS28). We show evidence that the regulation of GS28 by ORP7 does not occur at the level of transcription, but involves degradation of GS28 on proteasomes. Truncated ORP7 that lacks the GATE-16 binding region failed to affect GS28 stability, evidencing for specificity of the observed effect. Similar to ORP7 overexpression, treatment of cells with 25-hydroxycholesterol (25-OH) resulted in GS28 destabilization, which was potentiated by excess ORP7 and inhibited by ORP7 silencing. Overexpression of ORP7 led in most cells to formation of vacuolar structures positive for RFP-LC3, thus representing autophagic elements. Also GATE-16 was found in the vacuolar ORP7-positive elements, suggesting that excess ORP7 increases entrapment of GATE-16 in autophagosomes. Taken together, our results suggest that ORP7 negatively regulates GS28 protein stability via sequestration of GATE-16, and may mediate the effect of 25-OH on GS28 and Golgi function.
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http://dx.doi.org/10.1016/j.yexcr.2011.05.028DOI Listing
October 2011

Pinus massoniana bark extract selectively induces apoptosis in human hepatoma cells, possibly through caspase-dependent pathways.

Int J Mol Med 2010 May;25(5):751-9

The State Key Laboratory of Biocontrol and The Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen Zhongshan University, Guangzhou 510275, Guangdong Province, PR China.

Pinus massoniana bark extract (PMBE) is a mixture of flavonoids, whose antioxidant and apoptosis-inducing properties have been confirmed in vitro. In this study, the apoptotic effect and mechanism of PMBE in HepG2 human hepatoma cells were evaluated. PMBE exerted dose- and time-dependent cell growth inhibition on HepG2 cells, and selectively induced apoptosis without impact on normal liver L-02 cells. Apoptosis induced by PMBE in HepG2 cells was also confirmed by annexin-V/PI staining, transmission electron microscopy and sub-G1 phase accumulation. Moreover, PMBE also slightly blocked the cell cycle in the G2/M and S phases in HepG2 cells. The investigation of the mechanism by which PMBE induced apoptosis in HepG2 cells indicated that activation of extrinsic and intrinsic caspase, inhibition of NF-kappaB activation and decrease of the antiapoptotic protein Bcl-2 and the intact Bid protein were involved. Furthermore, the antitumor activity of PBME was demonstrated in vivo by a 42.88-69.94% reduction rate of tumor weight in H22 tumor-implanted mice. Taken together, these data indicate that PMBE selectively induces apoptosis in HepG2 cells through caspase-dependent pathways, and inhibits tumor growth in vivo, making it a potential candidate for anticancer therapeutics.
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http://dx.doi.org/10.3892/ijmm_00000401DOI Listing
May 2010

Effect of Pinus massoniana bark extract on IFN-gamma-induced ICAM-1 expression in HaCaT human keratinocytes.

J Ethnopharmacol 2009 Feb 3;122(1):48-53. Epub 2008 Dec 3.

The State Key Laboratory of Biocontrol and The Key Laboratory of Gene Engineering of Ministry of Education, School of Life Sciences, Sun Yat-sen (Zhongshan) University, Guangzhou 510275, Guangdong Province, PR China.

Aims Of The Study: Pinus massoniana bark extract (PMBE) with known anti-oxidant activity is comprised of various flavonoids including several bioactive compounds. We found that PMBE contains 1.27% taxifolin, a well-studied compound with known anti-inflammatory activity. Therefore, in the present study, we evaluated the effects of PMBE and taxifolin on intercellular adhesion molecule-1 (ICAM-1) expression.

Materials And Methods: PMBE and taxifolin were prepared. After HaCaT cells were pre-treatmented with PMBE and taxifolin, HaCaT cells were treatmented with 1000 U/ml IFN-gamma for 24 h.

Results: Treatment of HaCaT cells with 1000 U/ml IFN-gamma for 24 h markedly increased ICAM-1 expression. However, PMBE pre-treatment (40 microg/ml for 24 h) significantly inhibited IFN-gamma-induced ICAM-1 expression. In equal concentrations of taxifolin, PMBE-mediated inhibition of ICAM-1 mRNA and protein expression was greater than taxifolin mediated-inhibition, and the front on inhibition of ICAM-1 protein expression was 2.24-2.30-fold of the latter. When cells were treated with both compounds at a concentration of 40 microg/ml, PMBE-mediated inhibition of ICAM-1 mRNA was also greater than taxifolin-mediated inhibition and PMBE on inhibition of ICAM-1 protein expression was 2.60-3.00-fold the inhibition mediated by taxifolin.

Conclusions: PMBE including additional bioactive compounds may possibly synergize to inhibit transcription and translation of inducible ICAM-1expression and PMBE was greater than monomeric flavonoid taxifolin. These results indicate that PMBE exhibits great potential as a therapeutic treatment for inflammatory skin diseases.
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http://dx.doi.org/10.1016/j.jep.2008.11.024DOI Listing
February 2009
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