Publications by authors named "Shuangyan Wang"

37 Publications

Piccolo is essential for the maintenance of mouse retina but not cochlear hair cell function.

Aging (Albany NY) 2021 Apr 21;13(8):11678-11695. Epub 2021 Apr 21.

School of Life Science and Key Laboratory of the Ministry of Education for Experimental Teratology, Shandong University, Jinan 250100, China.

Piccolo is a presynaptic protein with high conservation among different species, and the expression of Piccolo is extensive in vertebrates. Recently, a small fragment of Piccolo (Piccolino), arising due to the incomplete splicing of intron 5/6, was found to be present in the synapses of retinas and cochleae. However, the comprehensive function of Piccolo in the retina and cochlea remains unclear. In this study, we generated knockout mice using CRISPR-Cas9 technology to explore the function of Piccolo. Unexpectedly, whereas no abnormalities were found in the cochlear hair cells of the mutant mice, significant differences were found in the retinas, in which two layers (the outer nuclear layer and the outer plexiform layer) were absent. Additionally, the amplitudes of electroretinograms were significantly reduced and pigmentation was observed in the fundoscopy of the mutant mouse retinas. The expression levels of Bassoon, a homolog of Piccolo, as well as synapse-associated proteins CtBP1, CtBP2, Kif3A, and Rim1 were down-regulated. The numbers of ribbon synapses in the retinas of the mutant mice were also reduced. Altogether, the phenotype of -/- mice resembled the symptoms of retinitis pigmentosa (RP) in humans, suggesting might be a candidate gene of RP and indicates knockout mice are a good model for elucidating the molecular mechanisms of RP.
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http://dx.doi.org/10.18632/aging.202861DOI Listing
April 2021

Surfactant cocamide monoethanolamide causes eye irritation by activating nociceptor TRPV1.

Br J Pharmacol 2021 Apr 10. Epub 2021 Apr 10.

State Key Laboratory of Natural Medicines and Department of TCM pharmacology, School of Traditional Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, China, 211198.

Background And Purpose: Cocamide monoethanolamide (CMEA) is commonly used as a surfactant-foam booster in cosmetic formulations. Upon contact with the eye or other sensitive skin areas, CMEA elicits sting and lasting irritation. We hypothesized a specific molecular interaction with TRPV1 by which CMEA caused eye irritation.

Experimental Approach: Eye irritancy is evaluated using eye-wiping test in rabbit and mouse. Intracellular Ca concentrations and action potentials are measured using Ca imaging and current clamp, respectively. Voltage clamp, site-direct mutagenesis and molecular modeling identify the binding pocket of CMEA on TRPV1.

Key Results: CMEA-induced eye irritation is ameliorated by selective ablation of TRPV1 and rodents exhibit much stronger responses to CMEA than rabbits. In trigeminal ganglion neurons, CMEA induces Ca influx and neuronal excitability, effects mitigated by TRPV1 inhibitor and absent in TRPV1 knockout neurons. In HEK-293 cell expressing TRPV1, CMEA increases whole-cell currents by increasing channel open probability (EC = 10.2 μM) without affecting TRPV2, 3, 4 and TRPA1 activities. Lauric acid monoethanolamide (LAMEA), the most abundant constituent in CMEA, is the most efficacious and potent TRPV1 activator. LAMEA binds to the capsaicin-binding pocket of TRPV1. Both rabbit TRPV1 that possesses the T550I variant and the human TRPV1 mutant exhibit much lower sensitivity to LAMEA.

Conclusions And Implication: CMEA directly activates TRPV1 to produce eye irritation, and rabbit, the traditional animal model used for eye irritancy test is a poor model for evaluating human eye irritants structurally related to CMEA. Our study identifies potential alternatives to CMEA as non-irritating surfactants.
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http://dx.doi.org/10.1111/bph.15491DOI Listing
April 2021

The Psychological Health Status of Healthcare Workers During the COVID-19 Outbreak: A Cross-Sectional Survey Study in Guangdong, China.

Front Public Health 2020 18;8:562885. Epub 2020 Sep 18.

Department of Geriatric Medicine, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, China.

The sudden outbreak of COVID-19 has caused mental stress on healthcare workers (HCW). This study aimed to assess their psychological health status at the peak of COVID-19 and to identify some coping strategies. A cross-sectional survey study was conducted during the outbreak of COVID-19. The survey was completed by 908/924 HCW (response rate 98.27%) in government-designated hospitals in Guangdong, China. A quality of life (QoL) scale, the Zung Self-Rating Anxiety Scale (SAS), and the Zung Self-Rating Depression Scale (SDS) were used to evaluate their psychological status. Logistic regression models were used to identify the occupational factors related to anxiety or depression. A total of 221 (24.34%) respondents had varying levels of anxiety, and 299 (32.93%) of them had depression. The mean SAS (42.9) and SDS (47.8) scores of HCW indicated that they were in the normal range for both anxiety and depression. Contact with COVID-19 cases or suspected cases, worry about suffering from COVID-19, worry about their family, and dismission during the COVID-19 period were significant work-related contributing factors to the psychological health problems of HCW (all p<0.01). The overall psychological health status of HCW in Guangdong, China, during the outbreak of COVID-19 was not overly poor. Updating and strengthening training in disease information, the provision of adequate medical supplies, and care about the life and health of medical staff and their family members may reduce their mental stress.
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http://dx.doi.org/10.3389/fpubh.2020.562885DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7530179PMC
September 2020

3'-O-Methylorobol Inhibits the Voltage-Gated Sodium Channel Nav1.7 with Anti-Itch Efficacy in A Histamine-Dependent Itch Mouse Model.

Int J Mol Sci 2019 Dec 1;20(23). Epub 2019 Dec 1.

State Key Laboratory of Natural Medicines and Jiangsu Provincial Key Laboratory for TCM Evaluation and Translational Development, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China.

An itch is a clinical complication that affects millions of patients. However, few treatment options are available. The voltage-gated sodium channel Nav1.7 is predominantly expressed in peripheral sensory neurons and is responsible for the rising phase of action potentials, thereby mediating nociceptive conduction. A gain-of-function mutation of Nav1.7 results in the hyperexcitability of sensory neurons and causes the inherited paroxysmal itch. Conversely, a monoclonal antibody that selectively inhibits Nav1.7 is able to effectively suppress the histamine-dependent itch in mice. Therefore, Nav1.7 inhibitors may possess the potential to relieve the itch. In the present study, using whole-cell voltage-clamp recordings, we demonstrated that 3'-O-methylorobol inhibited Na currents in Nav1.7-CHO cells and tetrodotoxin-sensitive Na currents in mouse dorsal root ganglion (DRG) neurons with IC (half-maximal inhibitory concentration) values of 3.46 and 6.60 μM, respectively. 3'-O-methylorobol also suppressed the tetrodotoxin-resistant Na currents in DRG neurons, though with reduced potency (~43% inhibition at 30 µM). 3'-O-methylorobol (10 µM) affected the Nav1.7 by shifting the half-maximal voltage (V) of activation to a depolarizing direction by ~6.76 mV, and it shifted the V of inactivation to a hyperpolarizing direction by ~16.79 mV. An analysis of 3'-O-methylorobol activity toward an array of itch targets revealed that 3'-O-methylorobol was without effect on histamine H receptor, TRPV1, TRPV3, TRPV4, TRPC4 and TRPM8. The intrathecal administration of 3'-O-methylorobol significantly attenuated compound 48/80-induced histamine-dependent spontaneous scratching bouts and the expression level of in the nuclei of spinal dorsal horn neurons with a comparable efficacy to that of cyproheptadine. Our data illustrated the therapeutic potential for 3'-O-methylorobol for histamine-dependent itching, and the small molecule inhibition of Nav1.7 may represent a useful strategy to develop novel therapeutics for itching.
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http://dx.doi.org/10.3390/ijms20236058DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6928743PMC
December 2019

Bacillus lacisalsi sp. nov., a moderately haloalkaliphilic bacterium isolated from a saline-alkaline lake.

Antonie Van Leeuwenhoek 2020 Jan 10;113(1):127-136. Epub 2019 Sep 10.

Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, People's Republic of China.

An alkaliphilic and moderately halophilic strain, designated YSP-3, characterised by optimal growth at pH 9.0 and at 8.0% (w/v) NaCl, was isolated from Yangshapao Lake, Jilin Province, China. Cells of this strain is Gram-positive, straight rods and form a central or sub-terminal ellipsoidal endospore. Phylogenetic analysis based on 16S rRNA gene sequences indicated that it was grouped in the genus Bacillus with Bacillus aurantiacus K1-5 and Bacillus populi FJAT-45347 as the close relative (97.5 and 97.2% 16S rRNA gene sequence similarity, respectively). Genomic relatedness between strain YSP-3 and its close relative was evaluated using average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity with the values of 70.3-85.1%, 19.7-20.1% and 71.5-71.6%, respectively. Comparative genomics analysis showed that strain YSP-3 has distinct amino acid bias and significantly differences from foreign invasion events during evolution relative to the reference strains. Cell-wall peptidoglycan contains meso-diaminopimelic acid. The predominant polar lipids are phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol. The predominant quinone is menaquinone-7. The major fatty acids of strain YSP-3 are anteiso-C, iso-C, iso-C, anteiso-C and Iso-C. DNA G + C content of strain YSP-3 is 48.3 mol%. Based on genomics analysis, physiological, biochemical and chemotaxonomic data, strain YSP-3 represent a novel species, for which the name Bacillus lacisalsi sp. nov. is proposed. The type strain is YSP-3 (  = ACCC 60365 = KCTC 33934).
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http://dx.doi.org/10.1007/s10482-019-01322-3DOI Listing
January 2020

Draft Genome Sequence of ACCC 60430, an Aerobic Halophilic Bacterium Isolated from a Salt Lake.

Microbiol Resour Announc 2019 Jun 6;8(23). Epub 2019 Jun 6.

Graduate School, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China

The haloalkaliphilic bacterium ACCC 60430, which grows optimally at 8.0% (wt/vol) Na and pH 9.0, was isolated from Keqin Lake in Qiqihaer, China. The draft genome includes 4,006 predicted genes and 3,784 coding sequences (CDSs). Genomic analysis showed that various genes may explain the mechanism of salt and alkali resistance.
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http://dx.doi.org/10.1128/MRA.00339-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6554603PMC
June 2019

Dehydrocrenatidine Inhibits Voltage-Gated Sodium Channels and Ameliorates Mechanic Allodia in a Rat Model of Neuropathic Pain.

Toxins (Basel) 2019 04 18;11(4). Epub 2019 Apr 18.

Jiangsu Key Laboratory of TCM Evaluation and Translational Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China.

(D. Don) Benn, a medical plant, is used in clinic to treat inflammation, pain, sore throat, and eczema. The alkaloids are the main active components in . In this study, we examined the analgesic effect of dehydrocrenatidine (DHCT), a β-carboline alkaloid abundantly found in in a neuropathic pain rat model of a sciatic nerve chronic constriction injury. DHCT dose-dependently attenuated the mechanic allodynia. In acutely isolated dorsal root ganglion, DHCT completely suppressed the action potential firing. Further electrophysiological characterization demonstrated that DHCT suppressed both tetrodotoxin-resistant (TTX-R) and sensitive (TTX-S) voltage-gated sodium channel (VGSC) currents with IC values of 12.36 μM and 4.87 µM, respectively. DHCT shifted half-maximal voltage (V) of inactivation to hyperpolarizing direction by ~16.7 mV in TTX-S VGSCs. In TTX-R VGSCs, DHCT shifted V of inactivation voltage to hyperpolarizing direction and V of activation voltage to more depolarizing potential by ~23.9 mV and ~12.2 mV, respectively. DHCT preferred to interact with an inactivated state of VGSCs and prolonged the repriming time in both TTX-S and TTX-R VGSCs, transiting the channels into a slow inactivated state from a fast inactivated state. Considered together, these data demonstrated that the analgesic effect of DHCT was likely though the inhibition of neuronal excitability.
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http://dx.doi.org/10.3390/toxins11040229DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6521113PMC
April 2019

Salipaludibacillus keqinensis sp. nov., a moderately halophilic bacterium isolated from a saline-alkaline lake.

Antonie Van Leeuwenhoek 2019 Jun 20;112(6):897-903. Epub 2019 Feb 20.

Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, People's Republic of China.

A novel Gram-stain positive, short rod, forming sub-terminal endospores of ellipsoidal shape, halophilic, alkaliphilic and aerobic bacterium, designated strain KQ-12, was isolated from a saline-alkaline lake in China, and characterised by a polyphasic taxonomic approach. The isolate grew at 4-40 °C (optimum, 25 °C), at pH 8.0-10.0 (pH 9.0) and in the presence of 0-16% (w/v) NaCl (8%). 16S rRNA gene sequence similarity of KQ-12 to species in the genera Salipaludibacillus ranged from 96.6 to 98.1%. Phylogenetic trees indicated that the strain should be assigned to the genus Salipaludibacillus. The polar lipids of KQ-12 were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified phospholipid and its major cellular fatty acids were anteiso-C, anteiso-C, iso-C, and C. The isoprenoid quinone was MK-7. These key chemotaxonomic properties also confirmed the affiliation of the strain to the genus Salipaludibacillus. However, some physiological, biochemical properties, low average nucleotide identity and low digital DNA-DNA hybridization relatedness values enabled the strain to be differentiated from closely related species of the genus Salipaludibacillus. Thus, KQ-12 can be classified as a novel species in the genus Salipaludibacillus, for which the name Salipaludibacillus keqinensis sp. nov. is proposed. The type strain is KQ-12 ( =  ACCC 60430   =  KCTC 33935).
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http://dx.doi.org/10.1007/s10482-018-01224-wDOI Listing
June 2019

Cloning, tissue distribution and methylation analyses of Lhx9 in Chinese tongue sole (Cynoglossus semilaevis).

Gene 2019 Apr 21;691:176-184. Epub 2018 Dec 21.

Yellow Sea Fisheries Research Institute, CAFS, Key Lab for Sustainable Development of Marine Fisheries, Ministry of Agriculture, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China. Electronic address:

Lhx9 is a LIM-homeodomain protein related to gonad development and sex reversal. In this study, we cloned and characterized CS-Lhx9 in the gonads of the Chinese tongue sole (Cynoglossus semilaevis). The full-length cDNA of CS-Lhx9 was 3123 bp, including an ORF of 1149 bp encoding 383 amino acids which contains two LIM domains and a homeobox domain. CS-Lhx9 transcripts were primarily observed in the testis of male and neomale at 1 yah, but nearly undetectable in the ovary. During the development of gonad, CS-Lhx9 exhibited an increasing trend and appeared to reach its peak value of expression in testis at 1 yah. In situ hybridization was performed in male and neomale gonads at 210 dah and 1 yah. The results showed strong CS-Lhx9 signals in the spermatids and spermatozoa (germ cells). Methylation level of CS-Lhx9 promoter was higher in female and lower in male and neomale gonads, showing a negative correlation with CS-Lhx9 expression.
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http://dx.doi.org/10.1016/j.gene.2018.12.025DOI Listing
April 2019

Improving short-term information spreading efficiency in scale-free networks by specifying top large-degree vertices as the initial spreaders.

R Soc Open Sci 2018 Nov 14;5(11):181137. Epub 2018 Nov 14.

School of Engineering and Technology, China University of Geosciences, Beijing, People's Republic of China.

The positive function of initially influential vertices could be exploited to improve spreading efficiency for short-term spreading in scale-free networks. However, the selection of initial spreaders depends on the specific scenes. The selection of initial spreaders needs to offer low complexity and low power consumption for short-term spreading. In this paper, we propose a selection strategy for efficiently spreading information by specifying a set of top large-degree vertices as the initially informed vertices. The essential idea behind the proposed selection strategy is to exploit the significant diffusion of the top large-degree vertices at the beginning of spreading. To evaluate the positive impact of initially influential vertices, we first build an information spreading model in the Barabási-Albert (BA) scale-free network; next, we design 54 comparative Monte Carlo experiments based on a benchmark strategy and the proposed selection strategy in different BA scale-free network structures. Experimental results indicate that (i) the proposed selection strategy can significantly improve spreading efficiency in the short-term spreading and (ii) both network size and number of hubs have a strong impact on spreading efficiency, while the number of initially informed vertices has a weak impact. The proposed selection strategy can be employed in short-term spreading, such as sending warnings or crisis information spreading or information spreading in emergency training or realistic emergency scenes.
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http://dx.doi.org/10.1098/rsos.181137DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6281924PMC
November 2018

Draft Genome Sequence of Bacillus sp. Strain YSP-3, a Halophilic, Alkaliphilic Bacterium Isolated from a Salt Lake.

Microbiol Resour Announc 2018 Aug 23;7(7). Epub 2018 Aug 23.

Graduate School of Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China.

The halophilic, alkaliphilic bacterium Bacillus sp. strain YSP-3 was isolated from a salt lake. It grows optimally at 8% (wt/vol) NaCl (pH 9.0). The draft genome is composed of 4,006 predicted genes. Genomic analysis showed that various genes are potentially involved in the adaptation mechanisms for osmotic stress and pH homeostasis.
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http://dx.doi.org/10.1128/MRA.00882-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6256447PMC
August 2018

Cyclin B1/CDK1-regulated mitochondrial bioenergetics in cell cycle progression and tumor resistance.

Cancer Lett 2019 02 24;443:56-66. Epub 2018 Nov 24.

Department of Radiation Oncology, School of Medicine, University of California at Davis, Sacramento, CA, USA. Electronic address:

A mammalian cell houses two genomes located separately in the nucleus and mitochondria. During evolution, communications and adaptations between these two genomes occur extensively to achieve and sustain homeostasis for cellular functions and regeneration. Mitochondria provide the major cellular energy and contribute to gene regulation in the nucleus, whereas more than 98% of mitochondrial proteins are encoded by the nuclear genome. Such two-way signaling traffic presents an orchestrated dynamic between energy metabolism and consumption in cells. Recent reports have elucidated the way how mitochondrial bioenergetics synchronizes with the energy consumption for cell cycle progression mediated by cyclin B1/CDK1 as the communicator. This review is to recapitulate cyclin B1/CDK1 mediated mitochondrial activities in cell cycle progression and stress response as well as its potential link to reprogram energy metabolism in tumor adaptive resistance. Cyclin B1/CDK1-mediated mitochondrial bioenergetics is applied as an example to show how mitochondria could timely sense the cellular fuel demand and then coordinate ATP output. Such nucleus-mitochondria oscillation may play key roles in the flexible bioenergetics required for tumor cell survival and compromising the efficacy of anti-cancer therapy. Further deciphering the cyclin B1/CDK1-controlled mitochondrial metabolism may invent effect targets to treat resistant cancers.
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http://dx.doi.org/10.1016/j.canlet.2018.11.019DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6759061PMC
February 2019

cDNA cloning, characterization, and expression analysis of the Rac1 and Rac2 genes from Cynoglossus semilaevis.

Fish Shellfish Immunol 2019 Jan 3;84:998-1006. Epub 2018 Nov 3.

Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology Yellow Sea Fisheries Research Institute, CAFS, Qingdao, 266071, China; Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Qingdao, 266071, China. Electronic address:

Rac1 and Rac2, belonging to the small Rho GTPase family, play an important role during the immune responses. In this study, a Rac1 homolog (CsRac1) and a Rac2 homolog (CsRac2) were cloned from the Cynoglossus semilaevis. The full-length of CsRac1 and CsRac2 cDNA was 1219 bp and 1047 bp, respectively. Both CsRac1 and CsRac2 contain a 579 bp open reading frame (ORF) which encoding a 192 amino acids putative protein. The predicted molecular weight of CsRac1 and CsRac2 was 21.41 kDa and 21.35 kDa, and their theoretical pI was 8.50 and 7.91, respectively. Sequence analysis showed that the conserved RHO domain was detected both from amino acid of CsRac1 and CsRac2. Homologous analysis showed that CsRac1 and CsRac2 share high conservation with other counterparts from different species. The CsRac1 and CsRac2 transcript showed wide tissue distribution, in which CsRac1 and CsRac2 exhibit the highest expression level in liver and gill, respectively. The expression level of CsRac1 and CsRac2 fluctuated in the liver and gill tissues at different time points after challenged by Vibrio harveyi. Specifically, CsRac1 and CsRac2 were significantly up-regulated at 48 h and 96 h post injection. Moreover, the knocking down of CsRac1 and CsRac2 in cell line (TSHKC) reduced the expression of CsPAK1, CsIL1-β and CsTNF-α. The present data suggests that CsRac1 and CsRac2 might play important roles in the innate immunity of half-smooth tongue sole.
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http://dx.doi.org/10.1016/j.fsi.2018.11.006DOI Listing
January 2019

Designing efficient hybrid strategies for information spreading in scale-free networks.

R Soc Open Sci 2018 Aug 1;5(8):180117. Epub 2018 Aug 1.

School of Engineering and Technology, China University of Geosciences, Beijing, People's Republic of China.

Designing a spreading strategy is one of the critical issues strongly affecting spreading efficiency in complex networks. In this paper, to improve the efficiency of information spreading in scale-free networks, we propose four hybrid strategies by combining two basic strategies, i.e. (i) the LS (in which information is preferentially spread from the large-degree vertices to the small-degree ones), and (ii) the SL (in which information is preferentially spread from the small-degree vertices to the large-degree ones). The objective in combining the two basic LS and SL strategies is to fully exploit the advantages of both strategies. To evaluate the spreading efficiency of the proposed four hybrid strategies, we first propose an information spreading model. Then, we introduce the details of the proposed hybrid strategies that are formulated by combining LS and SL. Third, we build a set of scale-free network structures by differently configuring the relevant parameters. In addition, finally, we conduct various Monte Carlo experiments to examine the spreading efficiency of the proposed hybrid strategies in different scale-free network structures. Experimental results indicate that the proposed hybrid strategies are effective and efficient for spreading information in scale-free networks.
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http://dx.doi.org/10.1098/rsos.180117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6124100PMC
August 2018

Massilia violaceinigra sp. nov., a novel purple-pigmented bacterium isolated from glacier permafrost.

Int J Syst Evol Microbiol 2018 Jul 31;68(7):2271-2278. Epub 2018 May 31.

MOE Key lab of Industrial Biocatalysis; Institute of Biochemical Engineering, Department of Chemical Engineering; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, PR China.

A Gram-stain-negative, motile and rod-shaped bacterium, designated strain B2, which can synthesize purple pigments of violacein and dexyoviolacein, was isolated from Tianshan glacier in Xinjiang, China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that it was grouped in the genus Massilia with Massilia glaciei B448-2, Massilia eurypsychrophila B528-3 and Massilia psychrophila B1555-1 as its closest relatives (98.2, 97.9 and 97.0 % 16S rRNA gene sequence similarity, respectively). Genomic relatedness between strain B2 and its closest relatives was evaluated using average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity, with values of 77.93-85.08 %, 22.4-23.4 % and 71.54-72.99 %, respectively. Q-8 was the major ubiquinone. The major fatty acids (>5 %) of strain B2 were summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C12 : 0 and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The major polar lipids included phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The DNA G+C content of strain B2 was 63.51 mol%. Based on genomic relatedness, physiological, biochemical and chemotaxonomic data, strain B2 (=CGMCC 1.6993=DSM 19531=KCTC 32446) is considered to represent a novel species within the genus Massilia, for which the name Massilia violaceinigra sp. nov. is proposed.
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http://dx.doi.org/10.1099/ijsem.0.002826DOI Listing
July 2018

HBME-1 expression in differentiated thyroid carcinoma and its correlation with the ultrasonic manifestation of thyroid.

Oncol Lett 2017 Dec 15;14(6):6505-6510. Epub 2017 Sep 15.

Longnan Hospital of Daqing City, Daqing, Heilongjiang 163453, P.R. China.

We investigated expression of the protein human bone marrow endothelial cell marker-1 (HBME-1) in differentiated thyroid carcinoma tissues, and analyzed its correlation with ultrasonic manifestation of thyroid. The immunohistochemistry (IHC) staining method was used to measure the expression of HBME-1 in patient with nodular goiter (control group), papillary differentiated thyroid carcinoma (papillary carcinoma group) and follicular differentiated thyroid carcinoma (follicular carcinoma group) to investigate the differences in expression of HBME-1. We further analyzed the correlation of the expression of HBME-1 in the papillary carcinoma group and the follicular carcinoma group with ultrasonic manifestation of thyroid. In both the papillary carcinoma group and the follicular carcinoma group, the levels of HBME-1 in affected tissues and the IHS score of HBME-1 expression were higher than those in the control group (p<0.05). In the papillary carcinoma group, the mean IHS score of HBME-1 expression in affected tissues was higher than in the follicular carcinoma group (p<0.05). There were no statistically significant differences in comparison to HBME-1 expression in affected tissues between the papillary carcinoma group and the follicular carcinoma group (p>0.05). Between the papillary carcinoma group and the follicular carcinoma group, the differences in the comparison of the nodule diameter, echo, shape, boundary, calcification and blood flow signal were statistically significant (p<0.05), but incidence rate of enlargement of cervical lymph nodules between the groups were not statistically significant (p>0.05). Among patients in the papillary carcinoma group, the comparison of the nodule diameter, echo, shape, boundary, calcification and blood flow signal between the HBME-1-positive patients and the HBME-1-negative patients showed no statistical significance (p>0.05), but in the nodules of HBME-1-positive patients, the proportion of blood flow signal was higher than that in the nodules of HBME-1-negative patients. Among patients in the follicular carcinoma group, there was no statistically significant differences in the comparison of ultrasonic manifestation of thyroid (p>0.05). Therefore, there are difference in HBME-1 expression and ultrasonic manifestations of thyroid in patients with papillary carcinoma and follicular differentiated thyroid carcinoma.
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http://dx.doi.org/10.3892/ol.2017.6971DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5678245PMC
December 2017

A Novel Prediction Model for Postmolar Gestational Trophoblastic Neoplasia and Comparison With Existing Models.

Int J Gynecol Cancer 2017 06;27(5):1028-1034

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

Objective: The aim of this study was to comparatively study a novel model and existing models of predicting postmolar gestational trophoblastic neoplasia (GTN).

Methods: Two hundred twenty-two patients with complete hydatidiform moles were enrolled retrospectively. A natural regression was noted in 195 patients (spontaneous regression group), whereas the remaining 27 patients entered postmolar GTN (postmolar GTN group). The upper limits of the 95% confidence interval of human chorionic gonadotropin (hCG) values and hCG regression rates were calculated aggregately from the spontaneous regression group. The 4 prediction models (weekly hCG regression curve and weekly hCG regression rate curve reported by previous studies; daily hCG regression curve and daily hCG regression rate curve pioneered by us) were then plotted. The individual hCG curve of the postmolar GTN group was plotted and compared with the prediction models, respectively. The individual hCG curve superimposing the prediction curve was considered showing an elevated risk of GTN.

Results: All patients with postmolar GTN were preidentified by daily hCG regression rate curve. The other 3 prediction models had a considerable rate of failure in identification. Mean diagnosis time of daily hCG regression rate curve was significantly lower (P = 0.008), with an average of 15.3 days gained compared with International Federation of Gynecology and Obstetrics criteria. Cochran Q test showed that daily hCG regression rate curve produced a significantly better performance in predicting postmolar GTN than weekly hCG regression curve (P = 0.01).

Conclusions: Our data showed that daily hCG regression rate curve gives a better prediction of postmolar GTN and might potentially enhance the monitoring of patients with molar pregnancy, especially those who could not adhere to International Federation of Gynecology and Obstetrics protocols. However, this preliminary research should not change current clinical practice until further validation is carried out.
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http://dx.doi.org/10.1097/IGC.0000000000000976DOI Listing
June 2017

Effect of transcutaneous acupoint electrical stimulation on propofol sedation: an electroencephalogram analysis of patients undergoing pituitary adenomas resection.

BMC Complement Altern Med 2016 Jan 27;16:33. Epub 2016 Jan 27.

Neuroscience Research Institute, Key Lab for Neuroscience, Peking University Health Science Center, Beijing, 100191, China.

Background: Transcutaneous acupoint electrical stimulation (TAES) as a needleless acupuncture has the same effect like traditional manual acupuncture. The combination of TAES and anesthesia has been proved valid in enhancing the anesthetic effects but its mechanisms are still not clear.

Methods: In this study, we investigated the effect of TAES on anesthesia with an electroencephalogram (EEG) oscillation analysis on surgery patients anesthetized with propofol, a widely-used anesthetic in clinical practice. EEG was continuously recorded during light and deep propofol sedation (target-controlled infusion set at 1.0 and 3.0 μg/mL) in ten surgery patients with pituitary tumor excision. Each concentration of propofol was maintained for 6 min and TAES was given at 2-4 min. The changes in EEG power spectrum at different frequency bands (delta, theta, alpha, beta, and gamma) and the coherence of different EEG channels were analyzed.

Results: Our result showed that, after TAES application, the EEG power increased at alpha and beta bands in light sedation of propofol, but reduced at delta and beta bands in deep propofol sedation (p < 0.001). In addition, the EEG oscillation analysis showed an enhancement of synchronization at low frequencies and a decline in synchronization at high frequencies between different EEG channels in either light or deep propofol sedation.

Conclusions: Our study showed evidence suggested that TAES may have different effects on propofol under light and deep sedation. TAES could enhance the sedative effect of propofol at low concentration but reduce the sedative effect of propofol at high concentration.
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http://dx.doi.org/10.1186/s12906-016-1008-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4729180PMC
January 2016

PtPLC, a pacifastin-related inhibitor involved in antibacterial defense and prophenoloxidase cascade of the swimming crab Portunus trituberculatus.

Fish Shellfish Immunol 2015 Mar 24;43(1):36-42. Epub 2014 Dec 24.

School of Marine Science, Ningbo University, Ningbo 315211, China.

Pacifastin-related inhibitor is a new family of serine protease inhibitors that regulate the proteolytic cascade in multiple biological processes. Contrary to the knowledge on the structure and inhibitory mechanism of pacifastin-like members in locust, very little is known about their functions. Here, we report the inhibitory activities in relation to the structural characteristics of pacifastin light chain (PtPLC) gene identified from the swimming crab Portunus trituberculatus. The mature PtPLC and five PLD-related domains with critical residues were expressed in Escherichia coli, and assayed for their activities. The recombinant PtPLC (rPtPLC) displayed inhibitory activities against trypsin and chymotrypsin in a dose dependent manner, with a preference for trypsin. Except for rPtPLC-D4, the other four rPtPLC-related domains could inhibit at least one of serine proteases. The enzyme specificity of PtPLC domains generally corresponded to the nature of the P1 residue at the reactive site. rPtPLC was able to inhibit the growth of Gram-negative bacteria Vibrio alginolyticus and Pseudomonas aeruginosa, but not the Gram-positive bacterium and fungus tested. Further phenoloxidase (PO) assay showed the rPtPLC could depress the crab proPO system activation in vitro, and lead to 72.8% inhibition of PO activity at the concentration of 9.11 μM. It also suppressed proPO activation induced by rPtcSP and rPtSPH1. As the first functional study of the recombinant PLC protein in crustaceans, the present results together indicate that PtPLC functions in the crab immune response possibly via inhibiting bacterial growth and regulating the proPO system.
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http://dx.doi.org/10.1016/j.fsi.2014.12.017DOI Listing
March 2015

Effects of hypertonic saline - hydroxyethyl starch and mannitol on serum osmolality, dural tension and hemodynamics in patients undergoing elective neurosurgical procedures.

Int J Clin Exp Med 2014 15;7(8):2266-72. Epub 2014 Aug 15.

Department of Anesthesiology, Beijing Sanbo Brain Hospital, Capital Medical University Beijing, China.

Objective: To investigate effect of equal volumes (250 ml) of 7.2% hypertonic saline - 6% hydroxyethyl starch (HS-HES) and 20% mannitol (M) on dural tension, serum osmolality and hemodynamics in patients during elective neurosurgical procedures.

Material And Methods: Forty ASA I-II patients scheduled for elective neurosurgical supratentorial procedures were randomly assigned to two groups. About 30 min before skull opening, patients received either HS-HES or M at infusion rate 750 ml/h. Dural tension score was used to evaluate the dural tension by neurosurgeons. Serum osmolality was tested at following time points: before, 125 ml infused, 250 ml infused, 30 min and 60 min after infusion. Hemodynamic variables were measured by FloTrac.

Results: Patients who received HS-HES had a significant decrease in dural tension scores (P < 0.05) and obtained more satisfactory brain relaxation for neurosurgeon (95% vs. 75%). In HS-HES group, the peak of serum osmolality occurred earlier and hyperosmolality lasted for longer time. Transient decrease in mean arterial pressure was observed in M group at 10 min after the start infusion (P < 0.01). Heart rate significantly decreased after HS-HES infusion, whereas no significant changes were observed in M group. In HS-HES group, stroke volume variation significantly decreased from 9.7 ± 3.5 at the initiation of infusion to 6.7 ± 2.4 at 30 min after the infusion and remained decreased more than 60 min while it decreased from 6.8 ± 3.1 to 5.3 ± 1.5 in M group. Moreover, urine output in HS-HES group from initiation to 60 min after the infusion was significantly less than those in M group.

Conclusion: HS-HES might be an alternative to mannitol in treatment of intracranial hypertension.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161579PMC
September 2014

Glycyrol suppresses collagen-induced arthritis by regulating autoimmune and inflammatory responses.

PLoS One 2014 18;9(7):e98137. Epub 2014 Jul 18.

Department of Biochemistry and Molecular Biology, Beijing Normal University, Gene Engineering and Biotechnology Beijing Key Laboratory, Beijing, P. R. of China.

Glycyrol is a natural compound extracted from Glycyrrhiza uralensis, first reported by us to be a new immunosuppressant. Here, we demonstrate its beneficial effect in collagen-induced arthritis (CIA) in mice, a model for rheumatoid arthritis (RA) in man, and we document the underlying mechanisms. Peroral administration of glycyrol significantly reduced clinical scores, alleviated cartilage and bone erosion and reduced levels of serum inflammatory cytokines. Glycyrol also decreased delayed-type hypersensitivity, improved carbon clearance and reduced acetic acid-induced capillary permeability. Furthermore, glycyrol decreased NF-κB and NFAT transcriptional activities and inhibited IL-2 expression. The therapeutic effect of glycyrol was associated with down-regulation of both autoimmune and inflammatory reactions. In addition, we demonstrated that glycyrol has minimal acute toxicity in mice. Therefore, we propose that glycyrol may hold promise for future treatment of RA.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0098137PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4103760PMC
November 2015

Comparison of 7.2% hypertonic saline - 6% hydroxyethyl starch solution and 6% hydroxyethyl starch solution after the induction of anesthesia in patients undergoing elective neurosurgical procedures.

Clinics (Sao Paulo) 2013 ;68(3):323-8

Department of Anesthesiology, Beijing Sanbo Brain Hospital, Capital Medical University, Beijing, China.

Objective: The ideal solution for fluid management during neurosurgical procedures remains controversial. The aim of this study was to compare the effects of a 7.2% hypertonic saline - 6% hydroxyethyl starch (HS-HES) solution and a 6% hydroxyethyl starch (HES) solution on clinical, hemodynamic and laboratory variables during elective neurosurgical procedures.

Methods: Forty patients scheduled for elective neurosurgical procedures were randomly assigned to the HS-HES group orthe HES group. Afterthe induction of anesthesia, patients in the HS-HES group received 250 mL of HS-HES (500 mL/h), whereas the patients in the HES group received 1,000 mL of HES (1000 mL/h). The monitored variables included clinical, hemodynamic and laboratory parameters. Chictr.org: ChiCTR-TRC-12002357

Results: The patients who received the HS-HES solution had a significant decrease in the intraoperative total fluid input (p<0.01), the volume of Ringer's solution required (p<0.05), the fluid balance (p<0.01) and their dural tension scores (p<0.05). The total urine output, blood loss, bleeding severity scores, operation duration and hemodynamic variables were similar in both groups (p>0.05). Moreover, compared with the HES group, the HS-HES group had significantly higher plasma concentrations of sodium and chloride, increasing the osmolality (p<0.01).

Conclusion: Our results suggest that HS-HES reduced the volume of intraoperative fluid required to maintain the patients undergoing surgery and led to a decrease in the intraoperative fluid balance. Moreover, HS-HES improved the dural tension scores and provided satisfactory brain relaxation. Our results indicate that HS-HES may represent a new avenue for volume therapy during elective neurosurgical procedures.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3611754PMC
http://dx.doi.org/10.6061/clinics/2013(03)oa07DOI Listing
January 2014

Crustins from eyestalk cDNA library of swimming crab Portunus trituberculatus: molecular characterization, genomic organization and expression analysis.

Fish Shellfish Immunol 2012 Oct 23;33(4):937-45. Epub 2012 Aug 23.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.

Crustins are cationic, cysteine-rich antimicrobial proteins, containing a single whey acidic protein (WAP) domain in the C-terminal end. Different from the reported Ptcrustin in the hemocytes, two novel crustin genes (PtCrustin2 and PtCrustin3) were cloned completely from the eyestalk cDNA library of Portunus trituberculatus in this study. All PtCrustins share the consensus cysteine motif and are considered as Type I crustins. Four exons and three introns are identified in genomic DNA sequence of PtCrustin3 while three exons and two introns in PtCrustin2. The mRNA transcripts of PtCrustin2 and PtCrustin3 are mainly detected in eyestalk and gills, but not in hemocytes. Although both PtCrutins are up-regulated after challenge of three microorganisms, PtCrustin3 seems to respond more quickly to microbial challenge than Ptcrustin2. Unlike most crustins, both recombinant PtCrustin2 and PtCrustin3 exhibit antibacterial activity against Gram-positive bacteria Micrococcus luteus and Staphyloccocus aureus and Gram-negative bacterium Pseudomonas aeruginosa. In addition, rPtCrustin2 is moderately active against yeast Pichia pastoris and rPtCrustin3 show significant activity against Gram-negative bacterium Vibrio alginolyticus. These results indicate that PtCrustin2 and PtCrustin3 are two novel crustins and play different roles in immune response of P. trituberculatus against microbial challenge.
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http://dx.doi.org/10.1016/j.fsi.2012.08.002DOI Listing
October 2012

Molecular cloning, expression pattern and antimicrobial activity of a new isoform of anti-lipopolysaccharide factor from the swimming crab Portunus trituberculatus.

Fish Shellfish Immunol 2012 Jul 12;33(1):85-91. Epub 2012 Apr 12.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.

A new isoform of anti-lipopolysaccharide factor (PtALF5) was cloned from eyestalk cDNA library of swimming crab Portunus trituberculatus. The full-length cDNA of PtALF5 was 1045 bp encoding 120 amino acids. PtALF5 shared lower amino acid similarity with other ALFs, yet it contained the conserved LPS-binding domain and was clearly member of the ALF family. The genomic fragment of PtALF5 contained two exons separated by one intron. Several tandem repeats were found in intron. The mRNA transcript of PtALF5 was predominantly expressed in the hemocytes but barely detectable in muscle. After challenge with Vibrio alginolyticus, a main pathogen causing high mortality in P. trituberculatus, the PtALF5 transcript in hemocytes showed a clear time-dependent response expression pattern with obvious decrease at 6 h and significant increase at 24 h. The recombinant PtALF5 protein revealed antimicrobial activity against Gram-negative bacteria V. alginolyticus and Pseudomonas aeruginosa, but did not inhibit the growth of the tested Gram-positive bacteria and fungus. These results together suggest that PtALF5 is a potent antibacterial protein against Gram-negative bacteria infection, and might function as a promising therapeutic agent in disease control of crab aquaculture.
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http://dx.doi.org/10.1016/j.fsi.2012.04.004DOI Listing
July 2012

First report of two thioredoxin homologues in crustaceans: molecular characterization, genomic organization and expression pattern in swimming crab Portunus trituberculatus.

Fish Shellfish Immunol 2012 May 16;32(5):855-61. Epub 2012 Feb 16.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.

Previously, we had reported two homologues of the thioredoxin (Trx) super-family (PtTrx1 and PtTrx2) identified from eyestalk and haemocytes cDNA library of swimming crab Portunus trituberculatus, respectively. It was the first report of two thioredoxin homologues from the same crustacean species. Here, we focused on the molecular characterization, genomic organization and expression pattern of PtTrx1 and PtTrx2. The full-length cDNA sequences of PtTrx1 and PtTrx2 were 739 and 1300 bp, encoding 105 and 133 amino acids, respectively. They both had a conserved CGPC active site and highly similar tertiary structures, which containing four β-sheets and four α-helixes. Specifically, PtTrx2 was encoded by a nuclear gene and its cellular localization was targeted to mitochondria by an N-terminal mitochondrial pre-sequence. Sequence analysis revealed PtTrx1 and PtTrx2 were encoded by different genomic locus. As the first analyzed genomic structure of PtTrxs in crustaceans, two introns with microsatellites were found in the open reading frame region of these genes. Quantitative real-time PCR analysis revealed the mRNA expression of PtTrx1 transcripts were mainly detected in gill, while, PtTrx2 in eyestalk and gill. The temporal expression levels of PtTrxs transcripts in haemocytes showed different expression patterns after challenge with Vibrio alginolyticus, Micrococcus luteus and Pichia pastoris. These results together indicate that PtTrxs should be involved in the responses to pathogen challenge of P. trituberculatus.
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http://dx.doi.org/10.1016/j.fsi.2012.02.013DOI Listing
May 2012

A new anti-lipopolysaccharide factor isoform (PtALF4) from the swimming crab Portunus trituberculatus exhibited structural and functional diversity of ALFs.

Fish Shellfish Immunol 2012 May 8;32(5):724-31. Epub 2012 Feb 8.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.

Anti-lipopolysaccharide factors (ALFs) are the potent antimicrobial peptides that can bind and neutralize lipopolysaccharide (LPS). In this study, a new isoform of the ALF homologs (PtALF4) was cloned from eyestalk cDNA library of swimming crab Portunus trituberculatus. PtALF4 shared lower amino acid similarity with other ALFs, yet it contained the conserved LPS-binding domain and was clearly member of the ALF family. The genomic sequence of PtALF4 consisted of three exons interrupted by two introns. Several tandem repeats were found in both introns. Unlike most ALFs expressed in hemocytes, PtALF4 transcript was predominantly detected in eyestalk. After challenge with Vibrio alginolyticus, the temporal expression level of PtALF4 transcript in hemocytes showed a clear time-dependent response expression pattern with two significant peaks. The recombinant proteins of PtALF1, PtALF3 and PtALF4 revealed different antimicrobial activities against bacteria or fungus. These results together suggest that PtALF isoforms might be potent immune effectors to provide multiple protective functions against invading bacteria or fungus in P. trituberculatus.
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http://dx.doi.org/10.1016/j.fsi.2012.01.021DOI Listing
May 2012

Identification and characterization of two novel types of non-clip domain serine proteases (PtSP and PtSPH1) from cDNA haemocytes library of swimming crab Portunus trituberculatus.

Fish Shellfish Immunol 2012 May 21;32(5):683-92. Epub 2012 Jan 21.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.

In our previous studies, five serine proteases containing clip domain were characterized from the swimming crab Portunus trituberculatus. To further investigate the characterization and function of serine proteases, one serine protease (PtSP) and one serine protease homolog (PtSPH1) without clip domain were identified from haemocytes cDNA library in this paper. They both possessed an SP or SP-like domain at the C-terminal. In contrast to PtSP, absence of Ser catalytic residue resulted in the loss of serine protease activity of PtSPH1. Phylogenetic analysis suggested either SPs or SPHs might not have a single origin in gene evolution. Six introns presented in PtSP genomic DNA with one uncommon splice site (GG) was discovered at exon 1/intron 1 boundary region. Four introns with common splice sites were found in PtSPH1 genomic DNA. RT-PCR results showed that PtSP mRNA was mainly distributed in haemocytes, gill and eyestalk, whereas PtSPH1 transcript was mainly expressed in stomach. PtSP showed slight increase during the first 48 h compared to control groups except 8 h point after Micrococcus luteus challenge. However, significant up-regulation was observed in the expression level of PtSPH1 challenged by Gram-negative bacteria Vibrio alginolyticus, Gram-positive bacteria M. luteus and fungi Pichia pastoris during the first 48 h. It indicates that PtSPH1 might be more sensitive to microorganism challenges compared with PtSP.
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http://dx.doi.org/10.1016/j.fsi.2012.01.014DOI Listing
May 2012

Three clip domain serine proteases (cSPs) and one clip domain serine protease homologue (cSPH) identified from haemocytes and eyestalk cDNA libraries of swimming crab Portunus trituberculatus.

Fish Shellfish Immunol 2012 Apr 18;32(4):565-71. Epub 2012 Jan 18.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.

Four genes including three clip domain serine proteases (PtcSP1, PtcSP2 and PtcSP3) and one clip domain serine protease homologue (PtcSPH) of the swimming crab Portunus trituberculatus (Decapoda: Brachyura: Portunidae) were characterized based on analysis of expressed sequence tags from haemocytes and eyestalk cDNA libraries. The relative four peptidases, which share high structural similarity to the clip-SPs of other arthropod species, appeared to possess a clip domain at the N-terminus and an enzymatically active serine protease domain at the C-terminus except PtcSPH for its second catalytic residue Asp. (D) replaced by Ala (A). Alignment among the four full-sequences showed that PtcSP2 and PtcSP3 had the highest identical score (58%) while the similarity of other sequences was lower than 24%. The mRNA transcripts of PtcSPs and PtcSPH could be detected widely in all the examined tissues with remarkable different expression levels. The temporal expressions of PtcSPs and PtcSPH demonstrated different time-dependent expression pattern post Vibrio alginolyticus, Micrococcus luteus, and Pichia pastoris challenge. Especially, the expression of PtcSPH transcripts showed greater change against V. alginolyticus compared with the other two microorganisms. These findings suggest that PtcSPs and PtcSPH play different roles in the antibacterial defence mechanism of P. trituberculatus crab.
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http://dx.doi.org/10.1016/j.fsi.2012.01.006DOI Listing
April 2012

Identification and characterization of a serine protease inhibitor (PtSerpin) in the swimming crab Portunus trituberculatus.

Fish Shellfish Immunol 2012 Apr 10;32(4):544-50. Epub 2012 Jan 10.

EMBL, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.

Serine protease inhibitors (Serpins) play a key role in diverse immune biological processes. A serine protease inhibitor (Serpin), namely PtSerpin, was identified from the haemocyte cDNA library of swimming crab Portunus trituberculatus. The full-length PtSerpin cDNA was 1593 bp, including an open reading frame (ORF) of 1227 bp encoding a polypeptide of 408 amino acids with estimated molecular mass of 45.048 kDa and theoretical isoelectric point of 7.23. Predicted tertiary structure of PtSerpin contained three β-sheets and nine α-helices. Multiple sequence alignment revealed that deduced amino acid sequence of PtSerpin shared the highest similarity with serpin SPI from green mud crab Scylla paramamosain (SpSerpin). Phylogenetic analysis supported PtSerpin and SpSerpin were closely related to serpins from Penaeus monodon and Daphnia pulex while other decapods formed a separate group. Although the mRNA transcripts of PtSerpin could be detected in all the examined tissues, the higher levels were present in haemocytes and gills which are the major organs respond to pathogenic microorganism. After challenged by Vibrio alginolyticus, Micrococcus luteus and Pichia pastoris, the temporal expression of PtSerpin gene in haemocytes showed different activation times against bacteria and fungi within the experimental period of 72 h. These findings suggest that PtSerpin is involved in the antibacterial defense mechanism of P. trituberculatus crab.
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http://dx.doi.org/10.1016/j.fsi.2012.01.002DOI Listing
April 2012