Publications by authors named "Jinxiu Huang"

26 Publications

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The intestinal microbiota contributes to the growth and physiological state of muscle tissue in piglets.

Sci Rep 2021 May 27;11(1):11237. Epub 2021 May 27.

Chongqing Academy of Animal Science, Rongchang, Chongqing, 402460, China.

Although the importance of the intestinal microbiota in host growth and health is well known, the relationship between microbiota colonization and muscle development is unclear. In this study, the direct causal effects of the colonization of gut microorganisms on the muscle tissue of piglets were investigated. The body weight and lean mass of germ-free (GF) piglets were approximately 40% lower than those of normal piglets. The deletion of the intestinal microbiota led to weakened muscle function and a reduction in myogenic regulatory proteins, such as MyoG and MyoD, in GF piglets. In addition, the blinded IGF1/AKT/mTOR pathway in GF piglets caused muscle atrophy and autophagy, which were characterized by the high expression of Murf-1 and KLF15. Gut microbiota introduced to GF piglets via fecal microbiota transplantation not only colonized the gut but also partially restored muscle growth and development. Furthermore, the proportion of slow-twitch muscle fibers was lower in the muscle of GF piglets, which was caused by the reduced short-chain fatty acid content in the circulation and impaired mitochondrial function in muscle. Collectively, these findings suggest that the growth, development and function of skeletal muscle in animals are mediated by the intestinal microbiota.
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http://dx.doi.org/10.1038/s41598-021-90881-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8160342PMC
May 2021

Neuroprotective effect of stroke pretreated MSCs against cerebral ischemia/reperfusion injury in rats.

World Neurosurg 2021 May 3. Epub 2021 May 3.

Department of General Medicine, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, China. Electronic address:

Background: Mesenchymal stem cells (MSCs) have been shown to enhance neurological recovery after stroke. In this study, a middle cerebral artery occlusion (MCAO) rat model was designed to assess the neuroprotective effects of stroke pretreated MSCs on cerebral ischemia/reperfusion (I/R) injury.

Method: The MSCs were isolated and cultured in medium with 10% FBS, normal control serum (NS), or stroke serum (SS). Then these MSCs were injected into each group rats (n=6) 1 day after MCAO and continue feeding for 28 days. A battery of behavioral tests, TTC staining, hematoxylin and eosin staining, ELISA, and TUNEL assay were used to assess neural injury. Moreover, in order to detected the enhancement of neuronal regeneration and angiogenesis, immunofluorescence and western blotting were performed to assess the expression of trophic factor and growth factor.

Result: After treatment, the behaviour was improvement significantly. And the infarct area, brain lesion, and apoptosis cells were significantly decreased in the SS-MSCs group than others. It also modulated the inflammation by attenuating inflammatory cytokines. In addition, the number of neurogenesis positive cells, the expression of trophic factors and growth factors were significantly higher in the SS-MSCs group than others. At the same time, FBS-MSCs and NS-MSCs showed differences in the expression of trophic factors and growth factors, but the results were not as well as SS-MSCs.

Conclusion: SS-MSCs administration after the reperfusion led to neuroprotection by improving pathological changes, behavioral improvement, neurogenesis, suppression of apoptosis and inflammatory as well as angiogenesis.
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http://dx.doi.org/10.1016/j.wneu.2021.04.114DOI Listing
May 2021

Introduction of Colonic and Fecal Microbiota From an Adult Pig Differently Affects the Growth, Gut Health, Intestinal Microbiota and Blood Metabolome of Newborn Piglets.

Front Microbiol 2021 4;12:623673. Epub 2021 Feb 4.

Chongqing Academy of Animal Science, Chongqing, China.

Microbiota transplantation is a rapid and effective method for changing and reshaping the intestinal microbiota and metabolic profile in humans and animals. This study compared the different influences of the introduction of fecal microbes and colonic microbes from a fat, adult pig in newborn pigs. Both colonic microbiota transplantation (CMT) and fecal microbiota transplantation (FMT) promoted growth and improved gut functions in suckling pigs up to weaning. FMT was more beneficial for body weight gain and body fat deposition in piglets, while CMT was more beneficial for intestinal health and mucosal immunity. 16S rDNA sequence analysis indicated that both CMT and FMT significantly increased the abundances of beneficial or functional bacteria, such as and genera, in the piglets, and reduced the abundances of harmful bacteria, such as . Blood metabolome analysis showed that transplantation, especially FMT, enhanced lipid metabolism in piglets. In addition, while CMT also changed amino acid metabolism and increased anti-inflammatory metabolites such as 3-indoleacetic acid and 3-indolepropionic acid in piglets, FMT did not. Of note, FMT damaged the intestinal barrier of piglets to a certain extent and increased the levels of inflammatory factors in the blood that are potentially harmful to the health of pigs. Taken together, these results suggested that intestinal and fecal microbiota transplantations elicited similar but different physiological effects on young animals, so the application of microbiota transplantation in animal production requires the careful selection and evaluation of source bacteria.
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http://dx.doi.org/10.3389/fmicb.2021.623673DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7889522PMC
February 2021

Lactobacillus Plantarum 299v Changes miRNA Expression in the Intestines of Piglets and Leads to Downregulation of LITAF by Regulating ssc-miR-450a.

Probiotics Antimicrob Proteins 2021 Jan 24. Epub 2021 Jan 24.

Chongqing Academy of Animal Sciences, Rongchang, Chongqing, 402460, China.

Lactiplantibacillus plantarum subsp. plantarum 299v (L. plantarum 299v) is one of the most important probiotic strains in animal health, but the molecular mechanisms of how it exerts health benefits remain unclear. The purpose of this study was to explore the changes in miRNA expression profiles in the intestinal tissues of piglets by L. plantarum 299v and to explore its possible molecular regulatory mechanism in intestinal function. Neonatal piglets were orally administered L. plantarum 299v daily from 1 to 20 days old, and high-throughput sequencing was conducted to analyse the changes in miRNA expression in the jejunum and ileum. The results showed that 370 known porcine miRNAs were identified from eight libraries. Five miRNAs (ssc-miR-21-5p, -143-3p, -194b-5p, -192, and -126-3p) were highly expressed in the intestinal tissues. There were 15 differentially expressed miRNAs between the control group and the L. plantarum group, and only miR-450a was expressed differentially in both intestinal tissues. KEGG analysis revealed that the target genes of the 15 differentially expressed miRNAs were involved in 37 significantly enriched pathways (P < 0.01). Then, quantitative polymerase chain reaction confirmed that the miRNA expression was corresponded well with those from the sequencing. Luciferase reporter assays verified that lipopolysaccharide-induced TNF-α factor is a target of miR-450a. Our results also showed L. plantarum 299v could influence intestinal function by changing the levels of cytokines via miRNA expression. This is the first study to analyse differential expression miRNA profiles in intestinal tissue after L. plantarum 299v treatment and investigate the molecular regulatory mechanism of functional miRNA.
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http://dx.doi.org/10.1007/s12602-021-09743-1DOI Listing
January 2021

Changes of Gut Microbiota and Its Correlation With Short Chain Fatty Acids and Bioamine in Piglets at the Early Growth Stage.

Front Vet Sci 2020 5;7:617259. Epub 2021 Jan 5.

Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Chongqing, China.

The change characteristics of intestinal microbial succession and the correlation with the production of two important types of bacterial metabolites (short chain fatty acids and bioamine) in piglets during the early stage were fully explored in this study. Six piglets from different litters with the same birth time were selected, weighted and euthanized at 1, 7, 14, 21, 28, 35, and 42 days of age. During this stage, the piglets grew quickly with gradual increases in blood levels of growth hormone and insulin, and in the intestinal developmental index and immunity. 16s rRNA analysis indicated the alpha diversity of colonic microbiome community was higher than ileum. However, the composition change in the ileal microbiota was more dramatic over time. genus was the dominant bacteria in piglets' ileum while and genera were the dominant bacteria in colon up to weaning. Gut bacterial community of the piglets showed obvious differences between the three different phases: newborn, before weaning, and post weaning. This was similar to the morphological change pattern of pigs' gut. Total SCFA content in the colon of pigs showed almost a 20-fold increase at day 42 compared to the value at day 1. The percentage of acetic acid among the total SCFAs dropped quickly from 74.5% at day 1 to 36.5% at day 42, while butyric acid and propionic acid showed significant increases at the stage. The histamine level increased and putrescine level decreased markedly in the colon with time while the amounts of total bioamines, tyramine and spermidine were devoid of changes. Dozens bacteria taxa showed highly correlations with SCFAs and bioamines. These findings provide an expanded view of the dynamic pig gut and gut microbiome at the important early growth stage.
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http://dx.doi.org/10.3389/fvets.2020.617259DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7813758PMC
January 2021

Identification of differentially expressed miRNAs after Lactobacillus reuteri treatment in the ileum mucosa of piglets.

Genes Genomics 2020 11 26;42(11):1327-1338. Epub 2020 Sep 26.

Chongqing Academy of Animal Sciences, Rongchang, Chongqing, 402460, China.

Background: Lactobacillus reuteri I5007 possesses many excellent probiotic characteristics in piglets. miRNA plays important role in host-microbiota interactions, but the mechanism by which L. reuteri I5007 regulates intestinal function through its influence on miRNA expression is unknown.

Objective: This study analyzed the miRNA expression patterns in the ileum mucosa tissue of piglets by L. reuteri I5007 treatment, aim to clarify its molecular mechanism for regulating intestinal function through miRNA.

Methods: Neonatal piglets were orally administered L. reuteri I5007 or a placebo daily starting on day 1, and differential expression of ileal miRNAs was analyzed at 10 and 20 days of age by small RNA sequencing.

Results: 361 known porcine miRNAs were identified, and ten miRNAs were highly expressed in the ileum mucosa in both treatments. Nineteen differentially expressed (DE) miRNAs were identified in response to L. reuteri treatment, and four DE miRNAs (ssc-miR-196a, -196b-5p, -1285 and -10386) were differentially expressed at both time points. The KEGG pathway analyses showed the targets of 19 DE miRNAs were involved in 63 significantly enriched pathways, including the PI3K-Akt and MAPK pathways, which were confirmed to play important roles in probiotic-host communication. L. reuteri I5007 exerted anti-inflammatory effects by influencing the levels of inflammatory cytokines. Suppressor of cytokine signalling 4 gene was the target gene of ssc-miR-196a/-196b-5p, overexpression of ssc-miR-196a/-196b-5p downregulated the mRNA expression of IL-1β and TNFα in IPEC-J2 cells.

Conclusion: Our study provides new insight into the role of miRNAs in the intestinal function of piglets after L. reuteri I5007 treatment.
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http://dx.doi.org/10.1007/s13258-020-00998-6DOI Listing
November 2020

Effects of Lactobacillus plantarum on the intestinal morphology, intestinal barrier function and microbiota composition of suckling piglets.

J Anim Physiol Anim Nutr (Berl) 2019 Nov 9;103(6):1908-1918. Epub 2019 Sep 9.

Chongqing Academy of Animal Sciences, Chongqing, China.

This study investigated the effect of Lactobacillus plantarum strain 299v on gut health in suckling piglets. Sixty newborn piglets were assigned to control and probiotic treatments, with three litters per treatment (ten piglets/litter). From days 1 to 20 of life, piglets were orally administered a placebo of 0.1% peptone or 1.0 ×  10 CFU L. plantarum 299v daily. Six piglets per treatment were sacrificed on day 20, and intestinal tissues (including duodenum, jejunum, ileum and colon) and the intestinal contents from colon segments were collected. The results demonstrated that piglets treated with L. plantarum 299v had a lower diarrhoea incidence than the controls. L. plantarum 299v administration significantly increased the ratio of the villus height to the crypt depth in the jejunum and ileum, as well as the mRNA expression of jejunal occludin and ileal zonula occludens 1 (ZO-1). The L. plantarum treatment also increased the mRNA abundance of porcine β-defensin 2 (pBD2) and pBD3 in the jejunum and ileum and of toll-like receptors (TLRs), such as TLR2, TLR4, TLR6 and TLR9 in the ileum, and significantly upregulated the mRNA abundances of ileal pBD1 and colonic TLR4. Additionally, the L. plantarum 299v treatment significantly changed the structure of the colonic microbiota, as evidenced by the obvious increases in the relative abundances of the phyla Firmicutes and Actinobacteria and of the genus Lactobacillus. Our findings indicate that L. plantarum 299v facilitates the gut health of suckling piglets, probably by improving the intestinal morphology and intestinal barrier function and by modifying the structure of the gut microbiota.
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http://dx.doi.org/10.1111/jpn.13198DOI Listing
November 2019

Comparison of LncRNA Expression Profiles during Myogenic Differentiation and Adipogenic Transdifferentiation of Myoblasts.

Int J Mol Sci 2019 Jul 30;20(15). Epub 2019 Jul 30.

Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Rongchang, Chongqing 402460, China.

Myoblasts could transdifferentiate into adipocytes or adipocyte-like cells, which have the capability of producing and storing intracellular lipids. Long-chain non-coding RNAs (lncRNAs) have many important physiological functions in eukaryotes, which include regulating gene expression, chromosome silencing, and nuclear transport. However, changes in the expression of lncRNAs in muscle cells during adipogenic transdifferentiation have not been investigated to date. Here, C2C12 myoblasts were seeded and then induced to undergo myogenic and adipogenic transdifferentiation. The expression profiles of lncRNAs in various differentiated cells were analyzed and then compared by digital gene expression (DGE) RNA sequencing. A total of 114 core lncRNAs from 836 differentially expressed lncRNAs in adipogenic cells were identified. Further investigation by in silico analysis revealed that the target genes of core lncRNAs significantly enriched various signaling pathways that were related to glucose and lipid metabolism and muscle growth. The gene was a potential regulator of adipogenesis in muscle cells. It showed the highest levels of expression in adipogenic cells, and the knocking down negatively influenced lipid deposition in transdifferentiated myoblasts. This study has identified the novel candidate regulators that may be assessed in future molecular studies on adipogenic conversion of muscle cells.
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http://dx.doi.org/10.3390/ijms20153725DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6695777PMC
July 2019

MicroRNA-425 controls lipogenesis and lipolysis in adipocytes.

Biochim Biophys Acta Mol Cell Biol Lipids 2019 05 26;1864(5):744-755. Epub 2019 Feb 26.

Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Rongchang 402460, China. Electronic address:

An increasing number of studies have demonstrated that some microRNAs participate in the regulation of growth and development of adipocytes. The present study shows that microRNA-425-5p (miR-425) is a novel strong regulator of adipogenesis and adipolysis in adipocytes. Forced expression of miR-425 in mice promoted body fat accumulation and the development of obesity due to high-fat intake, whereas silencing miR-425 prevented mice from being obese. Mechanically, the expression of miR-425 is controlled by PPARγ during the adipogenesis process in adipocytes. MiR-425 overexpression resulted in a reduction in the proliferation of 3t3-L1 pre-adipocytes but significantly accelerated cellular adipogenic differentiation. Mapk14, a negative regulator of adipogenesis, was predicted and confirmed as a real target gene of miR-425. Moreover, knocking down miR-425 remarkably intensified intracellular lipolysis and promoted lipid oxidation, which is related to the activation of AMPK, a monitor for intracellular energy balance. MiR-425 activated AMPK not only by decreasing cellular ATP concentrations but also by targeting the gene of Cab39, which is an upstream co-activator of AMPK. The findings of the present study suggest that miR-425 could control adipogenesis and adipolysis in adipocytes by simultaneously triggering multidirectional targets.
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http://dx.doi.org/10.1016/j.bbalip.2019.02.007DOI Listing
May 2019

Evaluation of MYORG mutations as a novel cause of primary familial brain calcification.

Mov Disord 2019 02 27;34(2):291-297. Epub 2018 Dec 27.

Department of Neurology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.

Background: Very recently, the MYORG gene was identified as a novel causative gene for autosomal-recessive primary familial brain calcification.

Objective: To investigate the clinical, genetic, and neuroradiological characteristics of primary familial brain calcification patients with biallelic MYORG mutations in China.

Methods: We collected clinical and neuroradiological data of 169 Chinese patients with primary familial brain calcification, including 151 sporadic patients and 18 patients from 13 families compatible with an autosomal-recessive mode of inheritance. Mutational analysis of MYORG was performed in the cohort.

Results: We identified four, including three novel, MYORG mutations segregating in four families with 5 patients: one nonsense mutation (c.1431C>A, p.Y477*), one missense mutation (c.687G>T, p.W229C), and two nonframeshift indels (c.348_349insCTGGCCTTCCGC, p.116_117insLAFR; c. 428_442delTGCACTTCTTCATCC, p.143_147delLHFFI). The 12-base-pair insertion, c.348_349insCTGGCCTTCCGC, was found in either homozygous or heterozygous state in 2 probands of our cohort and another Chinese primary familial brain calcification patient previously reported on in the literature. Haplotype analysis of our patients harboring the insertion indicated a founder effect in the ethnic Han Chinese population. To date, biallelic MYORG mutations have been reported in 17 patients (including our cohort). Most patients were symptomatic (13 of 17; 76.5%), and the most recurrent symptoms were movement disorders (10 of 17; 58.8%), cognitive decline (7 of 17; 41.2%), and cerebellar symptoms (6 of 17; 35.3%). All patients had calcifications on comprehensive cranial CT, most frequently located in the basal ganglia (17 of 17; 100%), cerebellum (17 of 17; 100%), subcortical white matter (14 of 17; 82.4%), and thalamus (13 of 17; 76.5%).

Conclusions: We confirmed MYORG as a novel causative gene for primary familial brain calcification and further expanded the mutational and phenotypic spectrum of MYORG-related primary familial brain calcification. © 2018 International Parkinson and Movement Disorder Society.
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http://dx.doi.org/10.1002/mds.27582DOI Listing
February 2019

Expressions and Regulatory Effects of P38/ERK/JNK Mapks in the Adipogenic Trans-Differentiation of C2C12 Myoblasts.

Cell Physiol Biochem 2017 18;44(6):2467-2475. Epub 2017 Dec 18.

Chongqing Academy of Animal Science, Rongchang, China.

Background/aims: Myoblasts and muscle satellite cells have the potential to transdifferentiate into adipocytes or adipocyte-like cells. Previous studies suggest that mitogen-activated protein kinase (MAPK) is critical to adipogenic trans-differentiation of muscle cells. ERK1/2, P38 and JNK are three major MAPK family members; their activation and regulatory functions during adipogenic trans-differentiation of myoblasts are investigated.

Methods: C2C12 myoblasts were cultured and induced for adipogenic trans-differentiation. Activation patterns of MAPKs were assayed using protein microarray and Western blot. Three specific MAPK blockers, U0126, SB20358 and SP600125, were used to block ERK1/2, P38 and JNK during trans-differentiation. Cellular adipogenesis was measured using staining and morphological observations of cells and expression changes in adipogenic genes.

Results: Inhibitors reduced phosphorylation of corresponding MAPK and produced unique cellular effects. Suppressing P38 promoted adipogenic trans-differentiation and intensified adipolytic metabolism in differentiated cells. However, inhibition of ERK1/2 had the opposite effects on adipogenesis and no effect on adipolysis. Blocking JNK weakly blocked trans-differentiation but stimulated adipolysis and induced apoptosis.

Conclusion: Three MAPKs participate in the regulation of myoblast adipogenic trans-differentiation by controlling adipogenic and adipolysis metabolism.
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http://dx.doi.org/10.1159/000486169DOI Listing
March 2018

Effects of Conjugated Linoleic Acid Supplementation on the Expression Profile of miRNAs in Porcine Adipose Tissue.

Genes (Basel) 2017 Oct 13;8(10). Epub 2017 Oct 13.

Chongqing Academy of Animal Sciences, Chongqing 402460, China.

Conjugated linoleic acids (CLAs) play a major role in adipocyte differentiation and lipid metabolism in animals. MicroRNAs (miRNAs) appear to be involved in many biological processes in adipose tissue. However, the specific influence on miRNAs by CLA supplementation in porcine adipose tissue remains unclear. Thus, we continuously added 1.5% CLA to the pig diet from the embryo stage to the finishing period and conducted a high-throughput sequencing approach to analyse the changes in adipose tissue miRNAs. We identified 283 known porcine miRNAs, and 14 miRNAs were differentially expressed in response to CLA treatment. A Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the targets of the 14 differentially expressed miRNAs were involved in the Wnt signalling pathway. The CLA treatment downregulated the gene expression of PPARγ, C/EBPα, FAS, and FATP1 in both subcutaneous and abdominal fat tissues; the analysis showed that ssc-miR-21 expression was significantly correlated with PPARγ expression (<0.05), and speculated that ssc-miR-21 might influence adipogenesis through PPARγ. In conclusion, our study analysed the miRNA profiles in porcine adipose tissues by CLA treatment, and demonstrated that miRNAs are important regulators of fat lipogenesis. This study provides valuable information for the molecular regulatory mechanism of CLA on adipose tissue.
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http://dx.doi.org/10.3390/genes8100271DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5664121PMC
October 2017

MicroRNA-224-5p regulates adipocyte apoptosis induced by TNFα via controlling NF-κB activation.

J Cell Physiol 2018 Feb 9;233(2):1236-1246. Epub 2017 Jun 9.

Chongqing Academy of Animal Science, Rongchang, Chongqing, China.

Tumor necrosis factor (TNF) α can induce cell apoptosis and activate nuclear transcription (NF)-κB in different cell types. Activated NF-κB further promotes or suppresses cellular apoptosis in different cases. The present study explored the effect of activated NF-κB on adipocyte apoptosis induced by TNFα and which microRNAs (miRNAs) were involved in the process. Our findings demonstrated that treatment of differentiated 3T3-L1 adipocytes with TNFα (20 ng/mL) rapidly activated NF-κB and induced moderate apoptosis. Pyrrolidinedithiocarbamic acid (PDTC, 60 µM), a specific NF-κB inhibitor, abated NF-κB activation that rendered the adipocytes vulnerable to TNFα-induced apoptosis. Dozens of miRNAs exhibited significant expression changes following TNFα treatment and the addition of PDTC. In which, miRNA-224-5p (miR-224) was up-regulated by TNFα exposure but down-regulated by PDTC addition. Furthermore, over-expression of miR-224 promoted NF-κB activation and prevented the adipocyte apoptosis induced by TNFα, while miR-224 deficiency showed the opposite effects. The TRAF-associated NF-κB activator (TANK) gene was identified as a direct target of miR-224 by computational and luciferase reporter assays. Additionally, silencing the TANK gene by the small interfering RNA similarly promoted NF-κB activation and attenuated the cellular apoptosis. In conclusion, these findings demonstrate that miR-224 plays an essential role in adipocyte apoptosis caused by TNFα through control of NF-κB activation via targeting the TANK gene.
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http://dx.doi.org/10.1002/jcp.25992DOI Listing
February 2018

Differential expression profile of miRNAs in porcine muscle and adipose tissue during development.

Gene 2017 Jun 8;618:49-56. Epub 2017 Apr 8.

Chongqing Academy of Animal Sciences, Chongqing 402460, China; Key Laboratory of Pig Industry Sciences, Ministry of Agriculture; Chongqing Key Laboratory of Pig Industry Sciences, Chongqing 402460, China.. Electronic address:

MicroRNAs (miRNAs) are a class of non-coding RNAs that play a crucial regulatory role in many biological processes. Previous studies have reported miRNAs that are associated with the growth, differentiation, and proliferation of myocytes and adipocytes in pigs. However, differences in the miRNA expression profiles between muscle and adipose tissues during porcine development are unknown. Muscle and adipose tissues are the two major organs that are crucial for dynamic energy balance in the development and metabolism. Identification of differential expression profile of miRNAs will be useful for understanding the regulatory role of miRNAs in growth, development and evolution of these two tissues, and the research results will provide theoretical basis to improve meat quality. Therefore, we applied Hiseq sequencing to profile miRNAs in muscle and adipose tissues during four development stage at 1, 30, 90 and 240-day-old to explore their regulatory patterns at critical growth stages of pigs. We slaughtered 6 pigs at each developmental stages (24 pigs in total), respectively, RNA of three individual pigs were pooled and duplicate samples at each time point were given to sequence. We obtained a total of 96 million clean reads, and identified 329 known miRNAs and 157 novel miRNAs from all the libraries. We detected 37 miRNAs that were differentially expressed between porcine muscle and adipose tissues; 17 miRNAs which differentially expressed at 30, 90 and 240-day-old were considered as core differentially expressed miRNAs, among them, three miRNAs (ssc-miR-128, -133a-5p, -489) were differentially expressed at all four stages. KEGG analysis revealed the target genes of 17 core differentially expressed miRNAs were involved in 27 significantly enriched pathways (P<0.01), including the MAPK, Wnt, TGF-beta, and Notch signaling pathways, which are known to regulate myogenesis or are related to lipogenesis. 8 and 11 miRNAs were differentially expressed in the muscle and adipose tissues, respectively, of 30, 90, and 240-day-old pigs compared with the tissues of 1-day-old pigs. We selected five miRNAs from 17 core differentially expressed miRNAs to validate the miRNA-seq results by RT-qPCR, the RT-qPCR expression results corresponded well with those from the miRNA-seq. This study analyzed differential expression miRNA profiles between porcine skeletal muscle and adipose tissues during development, and provided valuable information to investigate the regulation mechanism of functional miRNAs.
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http://dx.doi.org/10.1016/j.gene.2017.04.013DOI Listing
June 2017

Expression Pattern and Regulatory Role of microRNA-23a in Conjugated Linoleic Acids-Induced Apoptosis of Adipocytes.

Cell Physiol Biochem 2016 30;40(3-4):668-680. Epub 2016 Nov 30.

Chongqing Academy of Animal Science Rongchang, China.

Background/aims: Conjugated linoleic acids (CLAs) are known to induce apoptosis in adipocytes; however, the cellular mechanisms involved remained illdefined. We explored the different apoptotic induction effects of two CLA isomers on adipocytes and then investigated the expression and function of microRNAs (miRNAs) related to the apoptosis.

Methods: TUNEL and FCM assays were used to detect CLAs-induced adipocyte apoptosis. Microarrays were used to compare the differential expression of miRNAs. MiR-23a, a miRNA that showed significant changes in expression in the CLA-treated cells, was selected for the subsequent functional studies via over-expression and knock down in in vivo and in vitro experiments.

Results: C9, t11-CLA exhibited a stronger induction of apoptosis in the differentiated 3T3-L1 adipocytes than t10, c12-CLA. However, t10, c12-CLA could rapidly activate NF-κB, which may have caused their different apoptotic effects. MiR-23a was markedly down-regulated by the CLAs treatment and miR-23a over-expression attenuated CLA-induced apoptosis. Apoptosis protease-activating factor 1 (APAF1) was identified as a target gene of miR-23a. In an in vivo experiment endogenous miR-23a was down-regulated in mice fed with a mixture of both CLAs. The mice also exhibited less fat deposition and more apoptotic fat cells in adipose tissue. Moreover, endogenous miR-23a was suppressed in mice via intravenous injection with an antagomir which resulted in decreased body weight, increased number of apoptotic fat cells and increased APAF1 expression in adipose tissue.

Conclusion: Taken together, our results suggest that miR-23a plays a critical role in CLA-induced apoptosis in adipocytes via controlling APAF1 expression.
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http://dx.doi.org/10.1159/000452579DOI Listing
February 2017

Effects of MicroRNA-23a on Differentiation and Gene Expression Profiles in 3T3-L1 Adipocytes.

Genes (Basel) 2016 Oct 24;7(10). Epub 2016 Oct 24.

Chongqing Academy of Animal Science, Rongchang, Chongqing 402460, China.

MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate growth, development, and programmed death of cells. A newly-published study has shown that miRNA-23a could regulate 3T3-L1 adipocyte differentiation. Here, we identified miRNA-23a as a negative regulator of 3T3-L1 adipocyte differentiation again. Over-expression of miRNA-23a inhibited differentiation and decreased lipogenesis as well as down-regulated mRNA and protein expression of both peroxisome proliferator-activated receptor (PPAR) γ and fatty acid binding protein (FABP) 4, whereas knock down of miRNA-23a showed the opposite effects on differentiation as well as increasing the number of apoptotic cells. Additionally, digital gene expression profiling sequencing (DGE-Seq) was used to assay changes in gene expression profiles following alterations in the level of miR-23a. In total, over-expression or knock down of miRNA-23a significantly changed the expression of 313 and 425 genes, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these genes were mainly involved in the stress response, immune system, metabolism, cell cycle, among other pathways. Additionally, the signal transducer and activator of transcription 1 (Stat1) was shown to be a target of miRNA-23a by computational and dual-luciferase reporter assays that indicated Janus Kinase (Jak)-Stat signal pathway was implicated in regulating adipogenesis mediated by miRNA-23a in adipocytes.
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http://dx.doi.org/10.3390/genes7100092DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5083931PMC
October 2016

MicroRNA-199a Targets the Fatty Acid Transport Protein 1 Gene and Inhibits the Adipogenic Trans-Differentiation of C2C12 Myoblasts.

Cell Physiol Biochem 2016 26;39(3):1087-97. Epub 2016 Aug 26.

Chongqing Academy of Animal Science, Rongchang, China.

Background/aims: Muscle cells are able to trans-differentiate into adipocytes with adipogenesis induction. MicroRNAs (miRNAs), a class of small non-coding RNAs, widely participate in the regulation of growth and development of cells. However, the expression and regulatory role of miRNAs in the trans-differentiation of muscle cell are largely unknown.

Methods: C2C12 myoblasts were inducted to adipogenesis trans-differentiation and microarrays were used to assay the changes of expression profile of miRNAs. MiR-199a, a miRNA showed significant change in the trans-differentiation, was selected for the subsequent function study via over- expression and knock down.

Results: Dozens of miRNAs showed different changes followed the adipogenesis trans-differentiation of C2C12 cells. In which, miR-199a was decreased in the adipogenic cells and miR-199a over-expression inhibited the trans-differentiation and decreased lipid accumulation in the cells. Moreover, Fatty acid transport protein 1 (Fatp1), a major regulator of trans-membrane transportation and the oxidative metabolism of free fatty acids, was showed to be a target of miR-199a by computational and luciferase reporter assays. Additionally, Fatp1 knock-down by small interfering RNA had similar inhibitory effects on the trans-differentiation in C2C12 cells.

Conclusion: Our study reveals an important role for miR-199a in the regulation of adipogenic trans-differentiation in muscle cells via suppression of Fatp1 gene.
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http://dx.doi.org/10.1159/000447817DOI Listing
February 2017

Advance in spinal cord ischemia reperfusion injury: Blood-spinal cord barrier and remote ischemic preconditioning.

Life Sci 2016 Jun 7;154:34-8. Epub 2016 Apr 7.

Department of Anesthesiology, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, Hubei, China.

The blood-spinal cord barrier (BSCB) is the physiological and metabolic substance diffusion barrier between blood circulation and spinal cord tissues. This barrier plays a vital role in maintaining the microenvironment stability of the spinal cord. When the spinal cord is subjected to ischemia/reperfusion (I/R) injury, the structure and function of the BSCB is disrupted, further destroying the spinal cord homeostasis and ultimately leading to neurological deficit. Remote ischemic preconditioning (RIPC) is an approach in which interspersed cycles of preconditioning ischemia is followed by reperfusion to tissues/organs to protect the distant target tissues/organs against subsequent lethal ischemic injuries. RIPC is an innovation of the treatment strategies that protect the organ from I/R injury. In this study, we review the morphological structure and function of the BSCB, the injury mechanism of BSCB resulting from spinal cord I/R, and the effect of RIPC on it.
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http://dx.doi.org/10.1016/j.lfs.2016.03.046DOI Listing
June 2016

Comparative study on copper leaching from waste printed circuit boards by typical ionic liquid acids.

Waste Manag 2015 Jul 11;41:142-7. Epub 2015 Apr 11.

School of Chemistry and Environmental Science, Shaanxi University of Technology, Hanzhong, Shaanxi 723001, China.

Waste printed circuit boards (WPCBs) are attracting increasing concerns because the recovery of its content of valuable metallic resources is hampered by the presence of hazardous substances. In this study, we used ionic liquids (IL) to leach copper from WPCBs. [BSO3HPy]OTf, [BSO3HMIm]OTf, [BSO4HPy]HSO4, [BSO4HMim]HSO4 and [MIm]HSO4 were selected. Factors that affect copper leaching rate were investigated in detail and their leaching kinetics were also examined with the comparison of [Bmim]HSO4. The results showed that all six IL acids could successfully leach copper out, with near 100% recovery. WPCB particle size and leaching time had similar influences on copper leaching performance, while IL acid concentration, hydrogen peroxide addition, solid to liquid ratio, temperature, showed different influences. Moreover, IL acid with HSO4(-) was more efficient than IL acid with CF3SO3(-). These six IL acids indicate a similar behavior with common inorganic acids, except temperature since copper leaching rate of some IL acids decreases with its increase. The results of leaching kinetics studies showed that diffusion plays a more important role than surface reaction, whereas copper leaching by inorganic acids is usually controlled by surface reaction. This innovation provides a new option for recovering valuable materials such as copper from WPCBs.
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http://dx.doi.org/10.1016/j.wasman.2015.03.037DOI Listing
July 2015

Supplementation with conjugated linoeic acid decreases pig back fat deposition by inducing adipocyte apoptosis.

BMC Vet Res 2014 Jun 26;10:141. Epub 2014 Jun 26.

Chongqing Academy of Animal Science, Rongchang, Chongqing 402460, China.

Background: Conjugated linoleic acid (CLA), a C18 fatty acid with conjugated double bonds, has been shown to serve as a powerful anti-obesity agent by several research groups, although the precise mechanism remains elusive. Previous studies showed that CLA induced apoptosis in 3T3-L1 cells and in mice. The aim of this research was to clarify the role of CLA in adipocyte apoptosis in pigs, a relevant model for obesity research.

Results: Our results clearly show that back fat deposition of CLA-fed pigs was significantly lower than that of pigs in the control group. Moreover, some typical apoptotic cells were observed among the adipocytes of CLA-fed pigs. Furthermore, the CLA-fed pigs had reduced expression of the anti-apoptosis factor Bcl-2 and increased expression of the pro-apoptosis factors Bax and P53. Subsequently, increased cytochrome C was released from the mitochondria to the endochylema, and the caspase cascade was activated, resulting in cellular apoptosis. These results are consistent with the effects of Bcl-2 and Bax in regulating CLA-induced adipocyte apoptosis via the mitochondrial signaling pathway. However, the increased expression of tumor necrosis factor (TNF)-α and its receptor TNFR indicate that the effect of CLA might partly be through the death receptor signaling pathway in adipose cells.

Conclusions: Our study has demonstrated that CLA reduces pig body fat deposition, an outcome that is partly meditated by apoptosis of adipose cells, and that both the mitochondrial pathway and the death receptor pathway are involved in this effect.
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http://dx.doi.org/10.1186/1746-6148-10-141DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4074849PMC
June 2014

Leaching behavior of copper from waste printed circuit boards with Brønsted acidic ionic liquid.

Waste Manag 2014 Feb 15;34(2):483-8. Epub 2013 Nov 15.

Key Laboratory of Solid Waste Treatment and Resource Recycle, Ministry of Education, Southwest University of Science and Technology, Mianyang 621010, China.

In this work, a Brønsted acidic ionic liquid, 1-butyl-3-methyl-imidazolium hydrogen sulfate ([bmim]HSO4), was used to leach copper from waste printed circuit boards (WPCBs, mounted with electronic components) for the first time, and the leaching behavior of copper was discussed in detail. The results showed that after the pre-treatment, the metal distributions were different with the particle size: Cu, Zn and Al increased with the increasing particle size; while Ni, Sn and Pb were in the contrary. And the particle size has significant influence on copper leaching rate. Copper leaching rate was higher than 99%, almost 100%, when 1g WPCBs powder was leached under the optimum conditions: particle size of 0.1-0.25 mm, 25 mL 80% (v/v) ionic liquid, 10 mL 30% hydrogen peroxide, solid/liquid ratio of 1/25, 70°C and 2h. Copper leaching by [bmim]HSO4 can be modeled with the shrinking core model, controlled by diffusion through a solid product layer, and the kinetic apparent activation energy has been calculated to be 25.36 kJ/mol.
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http://dx.doi.org/10.1016/j.wasman.2013.10.027DOI Listing
February 2014

Effects of suspended common-scale and nanoscale particles on the survival, growth and reproduction of Daphnia magna.

Chemosphere 2013 Nov 11;93(10):2644-9. Epub 2013 Oct 11.

Department of Environmental Science and Engineering, College of Chemical Engineering, Huaqiao University, Xiamen 361021, China; Institute of Environmental and Resources Technology, Huaqiao University, Xiamen 361021, China.

Suspended particles are a natural component of aquatic ecosystems. This study provides a report on the survival, growth and reproduction of common-scale and nanoscale particles of Daphnia magna Straus exposed to five types of particles (i.e. KN (kaolinite), MN (montmorillonite), MNn (nanoscale MN), NP (natural particles), and NPn (nanoscale NP)). The results of the study show that the suspended particles elicited a dose-dependent toxicity in KN, MN and MNn, with the following toxicity pattern: MN>KN>MNn. On the contrary, NP and NPn did not show any harmful effects on the animals. Instead, NP and NPn, especially NPn, contributed to the survivorship of the animals. The animals were able to survive throughout the 21-day period of bioassays, and when the particle concentration reached 600 mg L(-1), they produced many neonates without any addition of food. This experimental results also indicated that the nanoscale particles were less toxic than the common-scale ones, both for MN and NP. Moreover, the results of the 21-day period of bioassays indicated that when the organisms were exposed to particles of different size scales, they showed different selection patterns for allocating resources. This may be due to the generation of different assimilation and digestion patterns.
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http://dx.doi.org/10.1016/j.chemosphere.2013.08.096DOI Listing
November 2013

S-adenosylmethionine-induced adipogenesis is accompanied by suppression of Wnt/β-catenin and Hedgehog signaling pathways.

Mol Cell Biochem 2013 Oct 30;382(1-2):59-73. Epub 2013 Jul 30.

State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, China Agricultural University, Beijing, 100193, China,

S-Adenosylmethionine (SAM) plays a crucial role as a methyl donor in various biological processes and has been previously shown to be involved in adipogenesis in skeletal muscle. This study was conducted to explore the mechanism of SAM inducing adipogenesis in skeletal muscle. Adipose precursor cells, 3T3-L1, and C2C12 cells, were induced into adipogenic differentiation by addition of SAM in MDI-differentiation media (0.5 mmol/L isobutylmethylxanthine, 1 μm/L dexamethasone, and 10 μg/mL insulin) to explore the role of SAM in promoting adipogenesis. Subsequently, cells were cultured with a medium containing SAM alone at the beginning of differentiation to test the relationship between SAM-induced adipogenesis and Wnt/β-catenin, and Hedgehog signaling pathways that control the cell commitment to adipogenic- or myogenic-differentiation. We found SAM possessed an additive effect with MDI in promoting adipogenesis of 3T3-L1 and C2C12 cells at the beginning of adipogenic differentiation. SAM could also individually induce cell adipogenesis in a dose-dependent manner. Moreover, the expression of Wnt/β-catenin and Hedgehog signals and their targets were suppressed by SAM (P < 0.05). These results demonstrate that SAM, as an increasingly accepted nutritional supplement, can initiate adipogenesis of adipose precursor cells derived from adipose and muscle tissues, a function at least partly correlated with the suppression of Wnt/β-catenin and Hedgehog pathways.
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http://dx.doi.org/10.1007/s11010-013-1718-3DOI Listing
October 2013

Efficacy of cytidine-5'-diphosp-bocholine combined with compound anisodine in the treatment of early optic nerve contusion.

Eye Sci 2012 Mar;27(1):37-40

Department of Ophthalmology, People's Hospital of Lianjiang, Lianjiang, China.

Purpose: To investigate the efficacy of anisodine combined with cytidine-5'-diphosp-bocholine (citicoline) in the treatment of early optic nerve contusion.

Methods: A total of 33 subjects eligible for inclusion were selected from 105 patients clinically diagnosed with optic nerve contusion. These patients were subsequently divided into the control group (n=16) and the intervention group (n=17). In the control group, the participants received therapy consisting of glucocorticoids, mannitol, vasodilators and vitamin B. The patients in the intervention group additionally received anisodine in combination with citicoline. The visual acuity was graded on a scale from 0 to 8.

Results: Prior to treatment, the 25th, 50th and 75th percentiles of visual acuity grade were 3, 4 and 6.75 for the controls, and 3, 4 and 6.5 for the patients in the intervention group. (P=0.97). After treatment, the 25th, 50th and 75th percentiles of visual acuity grade were 4, 6 and 7.75 in the control group, and 7, 7 and 8 in the intervention group. (P=0.046). A significant difference was observed in both control (P=0.005) and intervention groups (P=0.001) when comparing presenting visual acuity before and after treatment.

Conclusion: The combination of anisodine and citicoline with standard steroid and mannitol therapy appears to be effective in the treatment of early optic nerve contusion.
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http://dx.doi.org/10.3969/j.issn.1000-4432.2012.01.008DOI Listing
March 2012

Clinical study on interferon treatment of early scarring in filtering bleb.

Eye Sci 2011 Dec;26(4):197-200

Department of Ophthalmology, People's Hospital of Lianjiang, Lianjiang, Guangdong Province, China.

Purpose: To investigate the effectiveness of needle revision combined with subconjuctival injection of interferon α-2b in reversing early scarring of filtering blebs following trabeculectomy surgery.

Methods: Twenty-five glaucoma patients (31 eyes) who presented with scarred or encapsulated filtering bleb after glaucoma surgery underwent needle revision in combination with subconjuctival injection of interferon α-2b, and were followed for 12 months. Intraocular pressure (IOP) and filtering bleb morphology were observed post treatment.

Results: The mean time until scarring occurred was 21.0±7.4 days. The average time between recognition of bleb scarring and completion of needle revision was 2.2±0.8 days. The time interval between surgery and needle revision was inversely correlated with the time until needle revision (r = -0.694, P<0.001). The mean IOPs before and after needle revision were 24.2±2.7mmHg and 19.6±3.8mmHg, respectively (t = 5.916,P<0.001). At the 12-month follow-up visit, 18 eyes (58.1%) achieved complete success in IOP control, and 6 eyes (19.4%) had conditional success. The overall success rate for needling was thus 77.4%. Subconjunctival hemorrhage was observed in 4 eyes during the needle revision procedure. Punctate staining was found in the corneal epithelium of 2 eyes. Shallow anterior chamber (Grade I or II) was identified in 5 eyes.

Conclusion: Slit-lamp needle revision combined with subconjunctival injection of interferon α-2b may be efficacious in the treatment of early scarring of filtering blebs, is easy and safe to perform, and may be considered for more widespread application.
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http://dx.doi.org/10.3969/j.issn.1000-4432.2011.04.003DOI Listing
December 2011

The mRNA of lipin1 and its isoforms are differently expressed in the longissimus dorsi muscle of obese and lean pigs.

Mol Biol Rep 2011 Jan 1;38(1):319-25. Epub 2010 Apr 1.

State Key Laboratory of Animal Nutrition, College of Animal Science, China Agricultural University, No. 2. Yuanmingyuan West Road, Beijing, 100193, People's Republic of China.

Lipin1 has been documented to play an important role in adipogenesis. In the present study, the mRNA expression level of lipin1 and its isoforms in longissimus dorsi muscle were determined by semi-quantification RT-PCR in lean PIC and obese Rongchang pigs. Further, we determined mRNA expression for lipin1 and its two isoforms in Rongchang obese pigs which had either a high or low intramuscular fat content. We demonstrate for the first time that porcine lipin1 has two alternative forms, lipin-α and lipin-β. Unlike mice and humans where the lipin-β has 99 more nucleotides than lipin-α, we found that in swine, lipin-β has 108 more nucleotides than lipin-α. Our results indicate that the longissimus dorsi muscle of Rongchang obese pigs have a higher level of mRNA expression for lipin1 and its isoforms than PIC lean pigs. Furthermore, Rongchang pigs with higher intramuscular fat content had a higher lipin1 and lipin-β mRNA expression in longissimus dorsisi muscle than Rongchang pigs with lower intramuscular fat content (P<0.05), whereas no difference was seen in lipin-α mRNA expression between Rongchang pigs with high or low intramuscular fat. The ratio of lipin-β mRNA to lipin-α mRNA was also significantly different between Rongchang pigs distinguished by a high intramuscular fat content compared with those with low intramuscular fat (P<0.05). These data suggested that the lipin1 gene may have a crucial effect on body lipid accumulation in pigs, whereas the lipin-β isoform may play an important role in intramuscular fat deposition in obese pigs.
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http://dx.doi.org/10.1007/s11033-010-0110-6DOI Listing
January 2011