cDNA cloning and mRNA expression of heat shock protein 70 gene in blood clam Tegillarca granosa against heavy metals challenge

In this study, the full-length heat shock protein 70 of Tegillarca granosa was cloned from cDNA library by rapid amplification of cDNA end (RACE). The open reading frame (ORF) of heat shock protein 70 was 1968 bp, and it encoded a protein of 655 amino acids with a predicted molecular weight of 71.48 kDa and an isoelectric point of 5.25. Basic local alignment search tool (BLAST) analysis showed that the heat shock protein 70 of T. granosa shared high similarity with other species, supporting that it is a new member of heat shock protein family. Western blot analysis revealed that the generated polyclonal antibodies could specially detect native protein from whole cell lysate of T. granosa. The spatial distribution confirmed that the heat shock protein 70 was abundant in visceral mass, gill and haemocytes, and weakly in foot, mantle and adductor. Heavy metal pollutes such as lead (Pb2+), cadmium (Cd2+) and copper (Cu2+) could induce the gene expression in similar manners by quantitative real-time polymerase chain reaction (PCR). The present results indicate that heat shock protein 70 of T. granosa may be involved in environmental pollution challenges and should be considered as one of T. granosa promising molecular marker candidates.Keywords: Tegillarca granosa, heat shock protein 70, heavy metals, quantitative real-time polymerase chain reaction (PCR) African Journal of Biotechnology Vol. 12(18), pp. 2341-235

Molecular Characterization of a Novel Big Defensin from Clam Venerupis philippinarum

Antimicrobial peptides (AMPs) are important mediators of the primary defense mechanism against microbial invasion. In the present study, a big defensin was identified from Venerupis philippinarum haemocytes (denoted as VpBD) by RACE and EST approaches. The VpBD cDNA contained an open reading frame (ORF) of 285 bp encoding a polypeptide of 94 amino acids. The deduce amino acid sequence of VpBD shared the common features of big defensin including disulfide array organization and helix structure, indicating that VpBD should be a new member of the big defensin family. The mRNA transcript of VpBD was up-regulated significantly during the first 24 hr after Vibrio anguillarum challenge, which was 7.4-fold increase compared to that of the control group. Then the expression decreased gradually from 24 hr to 96 hr, and the lowest expression level was detected at 96 hr post-infection, which was still 3.9-fold higher than that of control. The mature peptide of VpBD was recombined in Escherichia coli and purified for minimum inhibitory concentration (MIC) determination. The rVpBD displayed broad-spectrum inhibitory activity towards all tested bacteria with the highest activity against Staphyloccocus aureus and Pseudomonas putida. These results indicated that VpBD was involved in the host immune response against bacterial infection and might contribute to the clearance of invading bacteria

Cloning and characterization of a sialic acid binding lectins (SABL) from Manila clam Venerupis philippinarum

Sialic acid binding lectin (SABL) is a member of immunoglobulin-like lectins family that are thought to promote cell cell interactions and regulate the functions of cells in the innate and adaptive immune systems through glycan recognition. In the present study, the full-length cDNA of SABL was identified from Manila clam Venerupis philippinarum (denoted as VpSABL) by cDNA library and RACE approaches. The cDNA of VpSABL consisted of a 5'terminal untranslated region (UTR) of 62 bp, a 3' UTR of 354 bp with a poly (A) tail, and an open reading frame (ORF) of 588 bp encoding a polypeptide of 195 amino acids with a typical C1q domain in the C-terminus. Multiple alignment analysis indicated that the deduced amino acid of VpSABL shared higher positive to other SABLs and C1q-contained proteins and should be adopted typical 10 beta-strand jelly-roll folding topology common to all C1q-TNF family. Spatial expression analysis indicated that mRNA transcript of VpSABL was predominantly detectable in tissues of mantle, hepatopancreas and gill, and to a lesser degree in the tissues of muscle and haemocytes. After challenged by Vibrio anguillarum, the mRNA level of VpSABL in overall haemocytes population was recorded by quantitative real-time RT-PCR. VpSABL mRNA was down-regulated in the first 24 h post-infection. Then, the expression level increased to the peak at 72 h and recovered to the original level at 96 h. All these results indicated that VpSABL was involved in the immune response against microbe infection and might be contributed to the recognition of bacterial pathogens. (C) 2011 Elsevier Ltd. All rights reserved.Sialic acid binding lectin (SABL) is a member of immunoglobulin-like lectins family that are thought to promote cell cell interactions and regulate the functions of cells in the innate and adaptive immune systems through glycan recognition. In the present study, the full-length cDNA of SABL was identified from Manila clam Venerupis philippinarum (denoted as VpSABL) by cDNA library and RACE approaches. The cDNA of VpSABL consisted of a 5'terminal untranslated region (UTR) of 62 bp, a 3' UTR of 354 bp with a poly (A) tail, and an open reading frame (ORF) of 588 bp encoding a polypeptide of 195 amino acids with a typical C1q domain in the C-terminus. Multiple alignment analysis indicated that the deduced amino acid of VpSABL shared higher positive to other SABLs and C1q-contained proteins and should be adopted typical 10 beta-strand jelly-roll folding topology common to all C1q-TNF family. Spatial expression analysis indicated that mRNA transcript of VpSABL was predominantly detectable in tissues of mantle, hepatopancreas and gill, and to a lesser degree in the tissues of muscle and haemocytes. After challenged by Vibrio anguillarum, the mRNA level of VpSABL in overall haemocytes population was recorded by quantitative real-time RT-PCR. VpSABL mRNA was down-regulated in the first 24 h post-infection. Then, the expression level increased to the peak at 72 h and recovered to the original level at 96 h. All these results indicated that VpSABL was involved in the immune response against microbe infection and might be contributed to the recognition of bacterial pathogens. (C) 2011 Elsevier Ltd. All rights reserved

A ferritin from Dendrorhynchus zhejiangensis with heavy metals detoxification activity.

Ferritin, an iron homeostasis protein, has important functions in transition and storage of toxic metal ions. In this study, the full-length cDNA of ferritin was isolated from Dendrorhynchus zhejiangensis by cDNA library and RACE approaches. The higher similarity and conserved motifs for ferritin were also identified in worm counterparts, indicating that it belonged to a new member of ferritin family. The temporal expression of worm ferritin in haemocytes was analyzed by RT-PCR, and revealed the ferritin could be induced by Cd(2+), Pb(2+) and Fe(2+). The heavy metal binding activity of recombinant ferritin was further elucidated by atomic force microscopy (AFM). It was observed that the ferritin protein could form a chain of beads with different size against three metals exposure, and the largest one with 35~40 nm in height was identified in the Cd(2+) challenge group. Our results indicated that worm ferritin was a promising candidate for heavy metals detoxification

Cloning and characterization of allograft inflammatory factor-1 (AIF-1) from manila clam Venerupis philippinarum

Allograft inflammatory factor-1 (AIF-1) is a 17 kDa interferon-gamma-inducible Ca2+-binding EF-hand protein that plays a significant role not only in different host responses to inflammatory stimuli, but in the whole host immune defense reaction. In the present study, the full-length cDNA of AIF-1 was identified from manila clam Venerupis philippinarum (denoted as VpAIF-1) by cDNA library and RACE approaches. The cDNA of vpAIF-1 consisted of a 5-terminal untranslated region (UTR) of 153 bp, a 3'UTR of 219 bp with a poly (A) tail, and an open reading frame (ORF) of 516 bp encoding a polypeptide of 171 amino acids with the putative molecular mass of 17 kDa. The deduced amino acid of VpAIF-1 shared two EF hand Ca2+-binding motifs like other AIF-1s. Phylogenetic analysis further indicated that VpAIF-1 had higher evolutional conservation to invertebrate than vertebrate counterparts and should be a new member of the AIF-1 protein family. Spatial expression analysis indicated that mRNA transcript of VpAIF-1 was found to be most abundantly expressed in the tissues of haemocytes, gills and hepatopancreas, weakly expressed in the tissues of mantle, muscle, and foot. After challenged by Vibrio anguillarum, the mRNA level of VpAIF-1 in overall haemocytes population was recorded by quantitative real-time RT-PcR. vpAIF-1 mRNA was down-regulated in the first 12 h post-infection. Then, the expression level increased to the peak at 72 h and recovered to the 48 h-level at 96 h. All these results indicated that VpAIF-1 was involved in the immune response against microbe infection and might be contributed to the clearance of bacterial pathogens. (C) 2010 Elsevier Ltd. All rights reserved.Allograft inflammatory factor-1 (AIF-1) is a 17 kDa interferon-gamma-inducible Ca(2+)-binding EF-hand protein that plays a significant role not only in different host responses to inflammatory stimuli, but in the whole host immune defense reaction. In the present study, the full-length cDNA of AIF-1 was identified from manila clam Venerupis philippinarum (denoted as VpAIF-1) by cDNA library and RACE approaches. The cDNA of vpAIF-1 consisted of a 5-terminal untranslated region (UTR) of 153 bp, a 3'UTR of 219 bp with a poly (A) tail, and an open reading frame (ORF) of 516 bp encoding a polypeptide of 171 amino acids with the putative molecular mass of 17 kDa. The deduced amino acid of VpAIF-1 shared two EF hand Ca(2+)-binding motifs like other AIF-1s. Phylogenetic analysis further indicated that VpAIF-1 had higher evolutional conservation to invertebrate than vertebrate counterparts and should be a new member of the AIF-1 protein family. Spatial expression analysis indicated that mRNA transcript of VpAIF-1 was found to be most abundantly expressed in the tissues of haemocytes, gills and hepatopancreas, weakly expressed in the tissues of mantle, muscle, and foot. After challenged by Vibrio anguillarum, the mRNA level of VpAIF-1 in overall haemocytes population was recorded by quantitative real-time RT-PcR. vpAIF-1 mRNA was down-regulated in the first 12 h post-infection. Then, the expression level increased to the peak at 72 h and recovered to the 48 h-level at 96 h. All these results indicated that VpAIF-1 was involved in the immune response against microbe infection and might be contributed to the clearance of bacterial pathogens. (C) 2010 Elsevier Ltd. All rights reserved

Prevalence of depressive symptoms among nurses in China: A systematic review and meta-analysis.

BACKGROUND:Depression is one of the most common mental disorders, profoundly impacting an individual's performance and quality of life. Due to their unique working conditions, nursing is counted among the occupational groups at high risk for developing depression. Because of the shortage of nursing resources in China, Chinese nurses suffer from heavy daily workloads more than those in many other countries. Therefore, this study aimed to evaluate the overall prevalence of depressive symptoms and analyse the potential risk factors of depressive symptoms in Chinese nurses. METHODS:A systematic literature search in PubMed, EMBASE, Web of Science, the Chinese BioMedical Literature Database (CBM), the China National Knowledge Infrastructure (CNKI), and the Weipu and Wanfang databases up to Dec 31st, 2019 was performed regarding the prevalence of depressive symptoms in Chinese nurses. Eligibility assessment and data extraction were performed independently by 2 researchers, and meta-analysis was used to synthesize the data. Heterogeneity was evaluated using Cochran's Q test and quantified using the I2 statistic. To explore the potential source of heterogeneity, subgroup analyses were also performed. In addition, both funnel plot and Egger's tests were adopted to assess publication bias. RESULTS:A total of 102 studies published from 1996 to 2019 covering 22 provinces were included for further analysis. The total number of participants was 52,592, with a range of 46 to 7205 per study. The overall prevalence of depressive symptoms in Chinese nurses was 43.83% (95%CI: 40.26%-47.42%), and 31.12% (95%CI: 27.30%-35.07%) were classified as mild degrees of depressive symptoms. The prevalence of depressive symptoms may be significantly affected by region, province or municipality and department marital status. Moreover, an increasing trend in the prevalence of depressive symptoms was observed in recent years. CONCLUSION:The results presented a high prevalence of depressive symptoms among Chinese nurses, which suggests interventional programmes by health decision-makers to improving the mental state of nurses is needed urgently, especially in nurses with high risk factors for depressive symptoms. Furthermore, the nationwide investigation of depressive symptoms prevalence should be performed with a standard diagnostic tool, which may be more useful for policy makers and planners

Two classes of glutathione S-transferase genes with different response profiles to bacterial challenge in Venerupis philippinarum

Start Research Fund; Ningbo University; MEL; NSFC [31101919]Glutathione S-transferase (GST) is major cytosolic detoxification enzymes involved in many pathological and physiological processes. In the present study, two classes of GSTs (VpGST-1 and VpGST-2) were cloned from Venerupis philippinarum haemocytes by cDNA library and RACE approaches. Sequence alignments and phylogenetic analysis together supported that they belonged to a new member of sigma and pi classes GSTs protein family, respectively. The expression profiles of these two genes under Vibrio anguillarum challenge were investigated by quantitative RT-PCR. The bacterial challenge could significantly up-regulate the mRNA expression of both VpGST-1 and VpGST-2 with larger amplitude in VpGST-2, and the feedback speed for VpGST-2 was more rapid than that of VpGST-1. The differences in the response to bacterial challenge indicated that they were functional diversity and probably played cooperative roles in mediating the Vibrio challenge in clam. (C) 2011 Elsevier Ltd. All rights reserved

Efficacy of Surgery Combined with Autologous Bone Marrow Stromal Cell Transplantation for Treatment of Intracerebral Hemorrhage

Bone marrow stromal cells (BMSCs) may differentiate into nerve cells under a certain condition; however, the clinical application for treating nervous system disease remains unclear. The aim is to assess the safety profile, feasibility, and effectiveness of surgery combined with autologous BMSCs transplantation for treating ICH. 206 ICH patients who had received surgical procedure were divided into transplantation (n=110) or control group (n=96). For transplantation group, BMSCs were injected into the perihemorrhage area in the base ganglia through an intracranial drainage tube 5.5 (3.01–6.89) days after surgery, followed by a second injection into the subarachnoid space through lumbar puncture 4 weeks later. Neurologic impairment and daily activities were assessed with National Institute Stroke Scale (NIHSS), Barthel index, and Rankin scale before transplantation and 6 months and 12 months after transplantation. Our results revealed that, compared with control group, NIHSS score and Rankin scale were both significantly decreased but Barthel index was increased in transplantation group after 6 months. Interestingly, no significant difference was observed between 12 months and 6 months. No transplantation-related adverse effects were investigated during follow-up assessments. Our findings suggest that surgery combined with autologous BMSCs transplantation is safe for treatment of ICH, providing short-term therapeutic benefits

Multiple alignment of VpBD with other known big defensins.

<p>Identical amino acids were in white letters with black background, and gray background indicated high levels of amino acid similarity.</p