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Bai F.,Southwest University | Luo L.,Southwest University | Huang X.,State Key Laboratory of Silkworm Genome Biology | Chen Y.,Southwest University | And 3 more authors.
Journal of Fisheries of China | Year: 2017

The experiment was conducted to study the feasibility of the application of silkworm pupal oil to GIFT Oreochromis niloticus feed. Seven isonitrogenous and isolipidic diets were formulated by adding 5.0% silkworm pupal oil (SPO), marine fish oil (MFO), tilapia oil (TO), pork lard (PL), chicken oil (CO), linseed oil (LO) and soybean oil (SO), respectively, to the basal diet to feed tilapia [initial body weight (47.51±0.52) g]. Each treatment contained three replicates with 20 fish per replicate and fish were fed for 56 days. The results showed that the final body weight, weight gain rate and specific growth rate of LO and SO groups were significantly higher than that of SPO and MFO groups, but showed no significant differences with that of TO, PL and CO groups. Feed conversion ratio of SPO group was significantly higher than that of other groups, except that of MFO group, while protein efficiency rate showed an opposite trend. Feed intake showed the lowest value in SPO group, and highest value in SO group. TO group presented the lowest viscerosomatic index, body length / body depth and intraperitoneal fat ratio and the highest condition factor. Hepatosomatic index in PL group was significantly higher than that of other groups except LO group. Intraperitoneal fat ratio in MFO group was significantly higher than that of other groups except CO group. Compared with the other groups, SPO could reduce the crude lipid in whole body lipid and tissue and triglyceride (TG), glucose and malonaldehyde contents in serum. MFO could reduce the crude lipid in whole body and enhance the acetyl-CoA carboxylase (ACC) activity in fish liver. PL groups presented lower TG and total cholesterol (TC) contents in serum and higher carnitine acyl transferase-I and ACC activities in liver. The highest TG, TC, low density lipoprotein cholesterol (LDL-C) contents and aspartate transaminase activity were observed in serum of CO group. LO and SO could significantly increase the lipid retention ratio and crude lipid content in whole body and muscle. SO significantly increased the high density lipoprotein cholesterol (HDL-C) and reduced LDL-C and TC contents in serum. SPO, LO and SO, respectively, could significantly increase highly unsaturated fatty acids, α-linolenic acid and linoleic acid contents in fish muscle. In conclusion, although fish growth performance is affected, SPO has advantages of decreasing body lipid, blood lipid and glucose, protecting fish health, and optimizing muscle fatty acids composition.


Datar I.,University of Toledo | Qiu X.,University of Toledo | Ma H.Z.,University of Toledo | Yeung M.,University of Toledo | And 8 more authors.
Oncotarget | Year: 2015

Accumulating evidence suggests that presence of macrophages in the tumor microenvironment add to the invasive and tumor-promoting hallmarks of cancer cells by secreting angiogenic and growth factors. RKIP is a known metastasis suppressor and interferes with several steps of metastasis. However, the mechanistic underpinnings of its function as a broad metastasis suppressor remain poorly understood. Here, we establish a novel pathway for RKIP regulation of metastasis inhibition through the negative regulation of RANTES/CCL5 thereby limiting tumor macrophage infiltration and inhibition of angiogenesis. Using a combination of loss- and gain-of- function approaches, we show that RKIP hinders breast cancer cell invasion by inhibiting expression of the CC chemokine CCL5 in vitro. We also show that the expression levels of RKIP and CCL5 are inversely correlated among clinical human breast cancer samples. Using a mouse allograft breast cancer transplantation model, we highlight that ectopic expression of RKIP significantly decreases tumor vasculature, macrophage infiltration and lung metastases. Mechanistically, we demonstrate that the inhibition of the CCL5 expression is the cause of the observed effects resulting from RKIP expression. Taken together, our results underscore the significance of RKIP as important negative regulator of tumor microenvironment.


PubMed | Kuwait University, National University of Singapore, University of Toledo and State Key Laboratory Of Silkworm Genome Biology
Type: Journal Article | Journal: Oncotarget | Year: 2015

Accumulating evidence suggests that presence of macrophages in the tumor microenvironment add to the invasive and tumor-promoting hallmarks of cancer cells by secreting angiogenic and growth factors. RKIP is a known metastasis suppressor and interferes with several steps of metastasis. However, the mechanistic underpinnings of its function as a broad metastasis suppressor remain poorly understood. Here, we establish a novel pathway for RKIP regulation of metastasis inhibition through the negative regulation of RANTES/CCL5 thereby limiting tumor macrophage infiltration and inhibition of angiogenesis. Using a combination of loss- and gain-of- function approaches, we show that RKIP hinders breast cancer cell invasion by inhibiting expression of the CC chemokine CCL5 in vitro. We also show that the expression levels of RKIP and CCL5 are inversely correlated among clinical human breast cancer samples. Using a mouse allograft breast cancer transplantation model, we highlight that ectopic expression of RKIP significantly decreases tumor vasculature, macrophage infiltration and lung metastases. Mechanistically, we demonstrate that the inhibition of the CCL5 expression is the cause of the observed effects resulting from RKIP expression. Taken together, our results underscore the significance of RKIP as important negative regulator of tumor microenvironment.


PubMed | Japan National Institute of Agrobiological Science, Institute of Biosciences &Applications, Indian Institute of Science, DNA Diagnostics Center and 3 more.
Type: | Journal: Scientific reports | Year: 2015

Despite more than 40 years of intense study, essential features of the silkmoth chorion (eggshell) are still not fully understood. To determine the precise structure of the chorion locus, we performed extensive EST analysis, constructed a bacterial artificial chromosome (BAC) contig, and obtained a continuous genomic sequence of 871,711 base pairs. We annotated 127 chorion genes in two segments interrupted by a 164kb region with 5 non-chorion genes, orthologs of which were on chorion bearing scaffolds in 4 ditrysian families. Detailed transcriptome analysis revealed expression throughout choriogenesis of most chorion genes originally categorized as middle, and evidence for diverse regulatory mechanisms including cis-elements, alternative splicing and promoter utilization, and antisense RNA. Phylogenetic analysis revealed multigene family associations and faster evolution of early chorion genes and transcriptionally active pseudogenes. Proteomics analysis identified 99 chorion proteins in the eggshell and micropyle localization of 1 early and 6Hc chorion proteins.


PubMed | Institute of Biosciences & Applications, Japan National Institute of Agrobiological Science, Indian Institute of Science, DNA Diagnostics Center and 3 more.
Type: | Journal: Scientific data | Year: 2015

The silkmoth chorion was studied extensively by F.C. Kafatos group for almost 40 years. However, the complete structure of the chorion locus was not obtained in the genome sequence of Bombyx mori published in 2008 due to repetitive sequences, resulting in gaps and an incomplete view of the locus. To obtain the complete sequence of the chorion locus, expressed sequence tags (ESTs) derived from follicular epithelium cells were used as probes to screen a bacterial artificial chromosome (BAC) library. Seven BACs were selected to construct a contig which covered the whole chorion locus. By Sanger sequencing, we successfully obtained complete sequences of the chorion locus spanning 871,711 base pairs on chromosome 2, where we annotated 127 chorion genes. The dataset reported here will recruit more researchers to revisit one of the oldest model systems which has been used to study developmentally regulated gene expression. It also provides insights into egg development and fertilization mechanisms and is relevant to applications related to improvements in breeding procedures and transgenesis.


Li G.-N.,Southwest University | Li G.-N.,State Key Laboratory of Silkworm Genome Biology | Xia X.-J.,Southwest University | Tang W.-C.,Southwest University | And 3 more authors.
Applied Microbiology and Biotechnology | Year: 2016

The silkworm (Bombyx mori L.) is an ideal model of Lepidoptera. However, the diversity and function of the intestinal microbiota in the gut of silkworm remain largely unknown. Changes in the intestinal microecology in fluoride-resistant strain T6 and fluoride-susceptible strain 734 of the silkworm in response to fluoride exposure were investigated. T6 and 734 were treated with 200 mg/kg fluoride (designated as T6-T and 734-T groups) and deionized water (designated as T6-C and 734-C groups). Culture-dependent approach revealed that the numbers of intestinal bacteria in the 734-T group significantly decreased compared with that in the 734-C group (4.8 ± 0.6 × 107 CFU/mL vs. 7.5 ± 0.7 × 107 CFU/mL; P < 0.05). Analyses of the intestinal content pH showed that the pH decreased in the 734-T group only. Additionally, SCFA concentrations significantly decreased in both treatment groups compared with the control groups. High-throughput sequencing indicated that the intestinal microbiota in the 734-T group was significantly more diverse than those in the other groups. The bacterial community was composed of two dominant groups (Firmicutes and Proteobacteria). Principal component analyses revealed a significant difference in the composition of the intestinal microbiota in the 734-T group compared with those in the other groups. Thaumarchaeota and Euryarchaeota were more abundant in the 734-T group, but they were less abundant in the other groups. This study enhances our understanding about the diversity and function of silkworm intestinal microbiota in response to fluoride exposure among silkworm strains with diverse resistance. © 2016 Springer-Verlag Berlin Heidelberg


Nie H.,State Key Laboratory of Silkworm Genome Biology | Liu C.,State Key Laboratory of Silkworm Genome Biology | Liu C.,Southwest University | Zhang Y.,State Key Laboratory of Silkworm Genome Biology | And 5 more authors.
PLoS ONE | Year: 2014

The ability to respond quickly and efficiently to transient extreme environmental conditions is an important property of all biota. However, the physiological basis of thermotolerance in different species is still unclear. Here, we found that the cot mutant showed a seizure phenotype including contraction of the body, rolling, vomiting gut juice and a momentary cessation of movement, and the heartbeat rhythm of the dorsal vessel significantly increases after hyperthermia. To comprehensively understand this process at the molecular level, the transcriptomic profile of cot mutant, which is a behavior mutant that exhibits a seizure phenotype, was investigated after hyperthermia (42°C) that was induced for 5 min. By digital gene expression profiling, we determined the gene expression profile of three strains (cot/cot ok/ok, +/+ ok/ok and +/+ +/+ ) under hyperthermia (42°C) and normal (25°C) conditions. A Venn diagram showed that the most common differentially expressed genes (DEGs, FDR,0.01 and log2 Ratio$1) were up-regulated and annotated with the heat shock proteins (HSPs) in 3 strains after treatment with hyperthermia, suggesting that HSPs rapidly increased in response to high temperature; 110 unique DEGs, could be identified in the cot mutant after inducing hyperthermia when compared to the control strains. Of these 110 unique DEGs, 98.18% (108 genes) were up-regulated and 1.82% (two genes) were downregulated in the cot mutant. KEGG pathways analysis of these unique DEGs suggested that the top three KEGG pathways were "Biotin metabolism," "Fatty acid biosynthesis" and "Purine metabolism," implying that diverse metabolic processes are active in cot mutant induced-hyperthermia. Unique DEGs of interest were mainly involved in the ubiquitin system, nicotinic acetylcholine receptor genes, cardiac excitation-contraction coupling or the Notch signaling pathway. Insights into hyperthermia-induced alterations in gene expression and related pathways could yield hints for understanding the relationship between behaviors and environmental stimuli (hyperthermia) in insects. © 2014 Nie et al.


Chai C.,Southwest University | Zhang Y.,Southwest University | Sun W.,Southwest University | Ding G.,Southwest University | And 5 more authors.
Gene | Year: 2014

In this report, we examined the gene expression related to carotenoid transport for a silkworm F1 hybrid with yellow cocoon generated by crossing two white-cocoon strains, Qiubai and 12-260. Our results showed that, in Qiubai, Cameo2, a transmembrane protein gene belonging to the CD36 family genes, was expressed normally in the silk gland, but no intact carotenoid-binding protein (CBP) mRNA (only the truncated CBP mRNA) was detected in the midgut. In 12-260, we detected the intact CBP mRNA expression in the midgut, but no Cameo2 expression in the silk gland. Regarding the F1 hybrid from crossing Qiubai and 12-260, both Cameo2 and intact CBP mRNA expressed normally in the silk gland and midgut. HPLC detection confirmed that in the F1 hybrid the carotenoids could be absorbed from dietary mulberry leaves through the midgut and transferred to silk gland via the hemolymph, which eventually colored cocoons into yellow. We also identified four CBP mRNA isoforms expressed in the midgut of the F1 hybrid, subsequently named as variants 5-8. Our results provide further evidences for the roles of Cameo2 and CBP in the formation of yellow cocoon of silkworm. © 2013 Elsevier B.V.


PubMed | Kuwait University, University of Toledo and State Key Laboratory Of Silkworm Genome Biology
Type: Journal Article | Journal: PloS one | Year: 2015

Raf Kinase Inhibitory Protein or RKIP was initially identified as a Raf-1 binding protein using the yeast 2-hybrid screen. RKIP inhibits the activation phosphorylation of MEK by Raf-1 by competitively inhibiting the binding of MEK to Raf-1 and thus exerting an inhibitory effect on the Raf-MEK-Erk pathway. RKIP has been identified as a metastasis suppressor gene. Expression of RKIP is low in cancer metastases. Although primary tumor growth remains unaffected, re- expression of RKIP inhibits cancer metastasis. Mechanistically, RKIP constrains metastasis by inhibiting angiogenesis, local invasion, intravasation, and colonization. The molecular mechanism of how RKIP inhibits these individual steps remains undefined. In our present study, using an unbiased PCR based screening and by analyzing DNA microarray expression datasets we observe that the expression of multiple metalloproteases (MMPs) including MMP1, MMP3, MMP10 and MMP13 are negatively correlated with RKIP expression in breast cancer cell lines and clinical samples. Since expression of MMPs by cancer cells is important for cancer metastasis, we hypothesize that RKIP may mediate suppression of breast cancer metastasis by inhibiting multiple MMPs. We show that the expression signature of RKIP and MMPs is better at predicting high metastatic risk than the individual gene. Using a combination of loss- and gain-of-function approaches, we find that MMP13 is the cause of RKIP-mediated inhibition of local cancer invasion. Interestingly expression of MMP13 alone is not sufficient to reverse the inhibition of breast cancer cell metastasis to the lung due to the expression of RKIP. We find that RKIP negatively regulates MMP13 through the Erk2 signaling pathway and the repression of MMP13 by RKIP is transcription factor AP-1 independent. Together, our findings indicate that RKIP inhibits cancer cell invasion, in part, via MMP13 inhibition. These data also implicate RKIP in the regulation of MMP transcription, suggesting a potential mechanism by which RKIP inhibits tumor progression and metastasis.


Li B.,Soochow University of China | Sun Q.,Soochow University of China | Yu X.,Soochow University of China | Xie Y.,Soochow University of China | And 6 more authors.
Environmental Toxicology | Year: 2015

It is known that exposure to organophosphorus pesticides (OP) including phoxim can produce oxidative stress, neurotoxicity, and greatly attenuate cocooning rate in the silkworm, Bombyx mori. Cerium treatment has been demonstrated to relieve phoxim-induced toxicity in B. mori; however, very little is known about the molecular mechanisms of silk gland injury due to OP exposure and protection of gland damage due to cerium pretreatment. The aim of this study was to evaluate silk gland damage and its molecular mechanisms in phoxim-induced silkworm toxicity and the protective mechanisms of cerium following exposure to phoxim. The results showed that phoxim exposure resulted in severe gland damage, reductions in protein synthesis and the cocooning rate of silkworms. Cerium (Ce) attenuated gland damage caused by phoxim, promoted protein synthesis, increased the antioxidant capacity of the gland and increased the cocooning rate of B. mori. Furthermore, digital gene expression data suggested that phoxim exposure led to significant up-regulation of 714 genes and down-regulation of 120 genes. Of these genes, 122 were related to protein metabolism, specifically, the down-regulated Ser2, Ser3, Fib-L, P25, and CYP450. Ce pretreatment resulted in up-regulation of 162 genes, and down-regulation of 141 genes, importantly, Ser2, Ser3, Fib-L, P25, and CYP333B8 were up-regulated. Treatment with CeCl3 + phoxim resulted in higher levels of Fib-L, P25, Ser2, Ser3, CAT, TPx, and CYP333B8 expression in the silk gland of silkworms. These findings indicated that Ce increased cocooning rate via the promotion of silk protein synthesis-related gene expression in the gland under phoxim-induced toxicity. These findings may expand the application of rare earths in sericulture. © 2014 Wiley Periodicals, Inc.

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