Publications by authors named "Yanjun Cui"

37 Publications

L. reuteri ZJ617 inhibits inflammatory and autophagy signaling pathways in gut-liver axis in piglet induced by lipopolysaccharide.

J Anim Sci Biotechnol 2021 Oct 13;12(1):110. Epub 2021 Oct 13.

The Key Laboratory of Molecular Animal Nutrition, Ministry of Education, College of Animal Science, Zhejiang University, Hangzhou, 310058, China.

Background: This study investigated the protective effects of L. reuteri ZJ617 on intestinal and liver injury and the underlying mechanisms in modulating inflammatory, autophagy, and apoptosis signaling pathways in a piglet challenged with lipopolysaccharide (LPS).

Methods: Duroc × Landrace × Large White piglets were assigned to 3 groups (n = 6/group): control (CON) and LPS groups received oral phosphate-buffered saline for 2 weeks before intraperitoneal injection (i.p.) of physiological saline or LPS (25 μg/kg body weight), respectively, while the ZJ617 + LPS group was orally inoculated with ZJ617 for 2 weeks before i.p. of LPS. Piglets were sacrificed 4 h after LPS injection to determine intestinal integrity, serum biochemical parameters, inflammatory signaling involved in molecular and liver injury pathways.

Results: Compared with controls, LPS stimulation significantly increased intestinal phosphorylated-p38 MAPK, phosphorylated-ERK and JNK protein levels and decreased IκBα protein expression, while serum LPS, TNF-α, and IL-6 concentrations (P < 0.05) increased. ZJ617 pretreatment significantly countered the effects induced by LPS alone, with the exception of p-JNK protein levels. Compared with controls, LPS stimulation significantly increased LC3, Atg5, and Beclin-1 protein expression (P < 0.05) but decreased ZO-1, claudin-3, and occludin protein expression (P < 0.05) and increased serum DAO and D-xylose levels, effects that were all countered by ZJ617 pretreatment. LPS induced significantly higher hepatic LC3, Atg5, Beclin-1, SOD-2, and Bax protein expression (P < 0.05) and lower hepatic total bile acid (TBA) levels (P < 0.05) compared with controls. ZJ617 pretreatment significantly decreased hepatic Beclin-1, SOD2, and Bax protein expression (P < 0.05) and showed a tendency to decrease hepatic TBA (P = 0.0743) induced by LPS treatment. Pretreatment of ZJ617 before LPS injection induced the production of 5 significant metabolites in the intestinal contents: capric acid, isoleucine 1TMS, glycerol-1-phosphate byproduct, linoleic acid, alanine-alanine (P < 0.05).

Conclusions: These results demonstrated that ZJ617 pretreatment alleviated LPS-induced intestinal tight junction protein destruction, and intestinal and hepatic inflammatory and autophagy signal activation in the piglets.
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http://dx.doi.org/10.1186/s40104-021-00624-9DOI Listing
October 2021

The potassium channel Kv1.3 as a therapeutic target for immunocytoprotection after reperfusion.

Ann Clin Transl Neurol 2021 Oct 7. Epub 2021 Oct 7.

Department of Pharmacology, School of Medicine, University of California, Davis, California, 95616, USA.

Objective: The voltage-gated potassium channel Kv1.3, which is expressed on activated, disease-associated microglia and memory T cells, constitutes an attractive target for immunocytoprotection after endovascular thrombectomy (EVT). Using young male mice and rats we previously demonstrated that the Kv1.3 blocker PAP-1 when started 12 h after reperfusion dose-dependently reduces infarction and improves neurological deficit on day 8. However, these proof-of-concept findings are of limited translational value because the majority of strokes occur in patients over 65 and, when considering overall lifetime risk, in females. Here, we therefore tested whether Kv1.3 deletion or delayed pharmacological therapy would be beneficial in females and aged animals.

Methods: Transient middle cerebral artery occlusion (tMCAO, 60 min) was induced in 16-week-old and 80-week-old male and female wild-type C57BL/6J and Kv1.3 mice. Stroke outcomes were assessed daily with the 14-score tactile and proprioceptive limp placing test and on day 8 before sacrifice by T2-weighted MRI. Young and old female mice were treated twice daily with 40 mg/kg PAP-1 starting 12 h after reperfusion. Microglia/macrophage activation and T-cell infiltration were evaluated in whole slide scans.

Results: Kv1.3 deletion provided no significant benefit in young females but improved outcomes in young males, old males, and old females compared with wild-type controls of the same sex. Delayed PAP-1 treatment improved outcomes in both young and old females. In old females, Kv1.3 deletion and PAP-1 treatment significantly reduced Iba-1 and CD3 staining intensity in the ipsilateral hemisphere.

Interpretation: Our preclinical studies using aged and female mice further validate Kv1.3 inhibitors as potential adjunctive treatments for reperfusion therapy in stroke by providing both genetic and pharmacological verification.
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http://dx.doi.org/10.1002/acn3.51456DOI Listing
October 2021

UPLC-MS/MS method for the determination of Lenvatinib in rat plasma and its application to drug-drug interaction studies.

J Pharm Biomed Anal 2021 Nov 2;206:114360. Epub 2021 Sep 2.

Department of Pharmacy, Hebei General Hospital, No. 348 West Heping Road, Shijiazhuang 050051, China. Electronic address:

Lenvatinib (LEN) is a multitargeted tyrosine kinase inhibitor registered for the first-line treatment of unresectable advanced hepatocellular carcinoma. Wuzhi capsule (WZC) is a traditional Chinese medicine preparation; it is used to decrease the aminotransferase level of the liver and protect liver function. Thus, patients with hepatocellular carcinoma (HCC) are potentially treated with a combination of LEN and WZC, but there is no information about the interaction between the two drugs. We developed a simple, rapid, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantitative determination of lenvatinib in rat plasma. Liquid-liquid extraction of plasma samples was carried out with ethyl acetate. Chromatographic separation of analyte was performed using gradient elution with acetonitrile and 0.1% formic acid water. The positive ion multi-response monitoring mode was used, and the target of the parent and daughter ions of LEN and IS were m/z 427.1→370 and m/z 432.1→370, respectively. All the validation projects were in accordance with the guidelines. Good linearity of 0.2-1000 ng/mL (r > 0.999) was achieved. The lower limit of quantification was 0.2 ng/mL. The precision and accuracy are acceptable. The method was successfully applied to pharmacokinetics and drug interaction analysis. The results show that WZC can significantly increase the C (maximum plasma concentration) and AUC (area under the concentration-time curve) of LEN. An UPLC -MS/MS method that can be used for studying drug-drug interaction as a valuable tool was developed in this study. Drug-drug interactions were observed between the WZC and LEN.
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http://dx.doi.org/10.1016/j.jpba.2021.114360DOI Listing
November 2021

Functionalized Attapulgite for the Adsorption of Methylene Blue: Synthesis, Characterization, and Adsorption Mechanism.

ACS Omega 2021 Aug 21;6(30):19586-19595. Epub 2021 Jul 21.

College of Science, Gansu Agricultural University, Lanzhou 730000, Gansu Province, China.

Attapulgite (denoted as APT, also called palygorskite) has been regarded as the green material in the "21st century world" and has attracted widespread attention due to its advantages of low cost, natural abundance, nontoxic nature, and environmental friendliness. However, the limited adsorption sites and surface charges of natural APT greatly hinder its application as an adsorbent in industrial fields. In this work, natural APT was modified with sodium humate (SA) via a facile one-step hydrothermal process to improve its adsorption capacity and systematically studied its ability to remove methylene blue (MB) from aqueous solutions. The effect of hydrothermal modification in the presence of SA on the microscopic structure, morphology, and physicochemical properties of APT was studied by field-emission scanning electron microscopy, Fourier transform infrared spectrometry, X-ray diffraction, and Brunauer-Emmett-Teller analyses. The adsorption properties of the modified APT toward MB were evaluated systematically. The results demonstrated that the modified APT has a high adsorption capacity of 227.27 mg/g and also shows a high removal rate up to 99.7% toward MB in a dye solution with an initial concentration of 150 mg/L, which was a 64.7% increase as compared to that of raw APT. The adsorption kinetics could be fitted to the pseudo-second-order model, while the adsorption isotherm could be well-described with the Langmuir model. It was concluded that electrostatic attraction, hydrogen-bonding interaction, and chemical association are the main driving force during the adsorption process.
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http://dx.doi.org/10.1021/acsomega.1c02111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8340108PMC
August 2021

Development, validation, and application of a simple UPLC-MS/MS method for simultaneous quantification of five traditional antipsychotics in human plasma.

Biomed Chromatogr 2021 Sep 6;35(9):e5143. Epub 2021 May 6.

College of Graduate Studies, Hebei Medical University, Shijiazhuang, China.

A UPLC-MS/MS method was developed to determine the levels of five traditional antipsychotics (APs) (chlorprothixene, perphenazine, fluphenazine, thioridazine, and promethazine) in human plasma with carbamazepine as the internal standard. Samples were extracted using simple liquid-liquid extraction (ethyl acetate/methyl tert-butyl ether, 2:3 v/v); then the analytes were subjected to gradient elution chromatography with a mobile phase composed of 0.1% formic acid in water and acetonitrile. The analytes were separated using a Waters XBridge BEH C column (100 × 2.1 mm, 2.5 μm). The linear ranges of chlorprothixene, perphenazine, fluphenazine, thioridazine, and promethazine are 2-250 ng/mL, r > 0.995. The limit of quantitation is 2 ng/mL, and the limit of detection is in the range of 0.1-0.5 ng/mL. The inter-day and intra-day relative standard deviations are less than 10%, and the relative errors are in the range of -5.70 to 7.20%. The recoveries of the five drugs are in the range of 70-109%. The results of methodology verification indicate that this method is simple, economical, sensitive, and suitable for the simultaneous quantification of five traditional APs in human plasma.
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http://dx.doi.org/10.1002/bmc.5143DOI Listing
September 2021

Carotid arterial hemodynamics in patients with essential hypertension of different dialectical types of traditional Chinese medicine.

Ann Palliat Med 2020 Nov;9(6):4097-4102

Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.

Background: To probe into the relationship between traditional Chinese medicine (TCM) different dialectical types of essential hypertension and carotid artery hemodynamics.

Methods: According to TCM, different dialectical types, 189 patients suffering from essential hypertension were divided into five type groups by two deputy chief physicians of TCM, according to TCM diagnosis and treatment guide for hypertension prepared by the Chinese society of TCM in 2011. A total of 189 patients were examined by transcranial color doppler (TCD) and compared with 47 normal subjects. The measuring target included middle cerebral artery (MCA) and anterior cerebral artery (ACA), recording the average blood flow velocity (Vm) and vascular pulse index (PI).

Results: In MCAVm and ACAVm, hyperpyrexia of liver syndrome showed unique advantages and positive correlation. In MCAPI and ACAPI, internal block because of phlegm dampness, stasis syndrome, deficiency of qi and blood syndrome, and internal block because of blood stasis syndrome had advantages and positive correlation, and the above three groups were not statistically significant.

Conclusions: Our study determined different dialectical types and objective material foundations of TCM in hypertension, correlating with carotid artery hemodynamics. Also, dialectical treatment would reduce the cerebrovascular accident.
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http://dx.doi.org/10.21037/apm-20-2060DOI Listing
November 2020

Preparation of natural rubber based semi-IPNs superabsorbent and its adsorption behavior for ammonium.

Int J Biol Macromol 2021 Jan 24;166:268-276. Epub 2020 Oct 24.

Institute of Agricultural Resources Chemistry and Application, College of Science, Gansu Agricultural University, Lanzhou 730070, China.

In this study, a natural rubber (NR) based amphiphilic semi-interpenetrating polymer network (semi-IPN) superabsorbent hydrogel was designed and synthesized with natural rubber-graft-poly (acrylic acid-co-acrylamide) [NR-g-P(AA-co-AM)] network and linear poly (diallyldimethyl ammonium chloride) (PDADMAC). Through a series of characterization and test, the structure, morphology, thermal properties, biodegradation, and swelling properties of NR-g-P(AA-co-AM)/PDADMAC were determined. Subsequently, NR-g-P(AA-co-AM)/PDADMAC was used for ammonium adsorption to remove ammonium nitrogen in aqueous solution. The adsorption behavior of the absorbent was also studied. Results showed that the maximum water absorbency of NR-g-P(AA-co-AM)/PDADMAC was 112.04 ± 6.55 g/g and water retention capacity of soil with the superabsorbent was 115.62 ± 2.08%. The NH adsorption quickly reached equilibrium and the maximum adsorption capacity was 13.02 mmol g calculated from Langmuir isotherm model. The results suggest that the product is efficient for ammonium removal and can be used as water-retaining agents.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.10.180DOI Listing
January 2021

Biophysical basis for Kv1.3 regulation of membrane potential changes induced by P2X4-mediated calcium entry in microglia.

Glia 2020 11 11;68(11):2377-2394. Epub 2020 Jun 11.

Department of Pharmacology, University of California, Davis, California, USA.

Microglia-mediated inflammation exerts adverse effects in ischemic stroke and in neurodegenerative disorders such as Alzheimer's disease (AD). Expression of the voltage-gated potassium channel Kv1.3 is required for microglia activation. Both genetic deletion and pharmacological inhibition of Kv1.3 are effective in reducing microglia activation and the associated inflammatory responses, as well as in improving neurological outcomes in animal models of AD and ischemic stroke. Here we sought to elucidate the molecular mechanisms underlying the therapeutic effects of Kv1.3 inhibition, which remain incompletely understood. Using a combination of whole-cell voltage-clamp electrophysiology and quantitative PCR (qPCR), we first characterized a stimulus-dependent differential expression pattern for Kv1.3 and P2X4, a major ATP-gated cationic channel, both in vitro and in vivo. We then demonstrated by whole-cell current-clamp experiments that Kv1.3 channels contribute not only to setting the resting membrane potential but also play an important role in counteracting excessive membrane potential changes evoked by depolarizing current injections. Similarly, the presence of Kv1.3 channels renders microglia more resistant to depolarization produced by ATP-mediated P2X4 receptor activation. Inhibiting Kv1.3 channels with ShK-223 completely nullified the ability of Kv1.3 to normalize membrane potential changes, resulting in excessive depolarization and reduced calcium transients through P2X4 receptors. Our report thus links Kv1.3 function to P2X4 receptor-mediated signaling as one of the underlying mechanisms by which Kv1.3 blockade reduces microglia-mediated inflammation. While we could confirm previously reported differences between males and females in microglial P2X4 expression, microglial Kv1.3 expression exhibited no gender differences in vitro or in vivo. MAIN POINTS: The voltage-gated K channel Kv1.3 regulates microglial membrane potential. Inhibition of Kv1.3 depolarizes microglia and reduces calcium entry mediated by P2X4 receptors by dissipating the electrochemical driving force for calcium.
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http://dx.doi.org/10.1002/glia.23847DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7540709PMC
November 2020

Lactobacillus rhamnosus GG Attenuates Lipopolysaccharide-Induced Inflammation and Barrier Dysfunction by Regulating MAPK/NF-κB Signaling and Modulating Metabolome in the Piglet Intestine.

J Nutr 2020 05;150(5):1313-1323

College of Animal Science, Zhejiang University, The Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Hangzhou, China.

Background: Probiotic Lactobacillius rhamnosus GG (LGG) shows beneficial immunomodulation on cultured cell lines in vitro and in mouse models.

Objective: The aim was to investigate the effects of LGG on intestinal injury and the underlying mechanisms by elucidating inflammatory signaling pathways and metabolomic response to LPS stimulation in the piglet intestine.

Methods: Piglets (Duroc × Landrace × Large White, including males and female; 8.6 ± 1.1 kg) aged 28 d were assigned to 3 groups (n = 6/group): oral inoculation with PBS for 2 wk before intraperitoneal injection of physiological saline [control (CON)] or LPS (25 μg/kg body weight; LPS) or oral inoculation with LGG for 2 wk before intraperitoneal injection of LPS (LGG+LPS). Piglets were killed 4 h after LPS injection. Systemic inflammation, intestinal integrity, inflammation signals, and metabolomic characteristics in the intestine were determined.

Results: Compared with CON, LPS stimulation significantly decreased ileal zonula occludens 1 (ZO-1; 44%), claudin-3 (44%), and occludin (41%) expression; increased serum diamineoxidase (73%), D-xylose (19%), TNF-α (43%), and IL-6 (55%) concentrations; induced p38 mitogen-activated protein kinase (p38 MAPK; 85%), extracellular signal-regulated kinase (ERK; 96%), and NF-κB p65 phosphorylation (37%) (P < 0.05). Compared with LPS stimulation alone, LGG pretreatment significantly enhanced the intestinal barrier by upregulating expressions of tight junction proteins (ZO-1, 73%; claudin-3, 55%; occludin, 67%), thereby decreasing serum diamineoxidase (26%) and D-xylose (28%) concentrations, and also reduced serum TNF-α expression (16%) and ileal p38 MAPK (79%), ERK (43%) and NF-κB p65 (37%) phosphorylation levels (P < 0.05). Metabolomic analysis showed clear separation between each group. The concentrations of caprylic acid [fold-change (FC) = 2.39], 1-mono-olein (FC = 2.68), erythritol (FC = 4.62), and ethanolamine (FC = 4.47) significantly increased in the intestine of LGG + LPS piglets compared with the LPS group (P < 0.05).

Conclusions: These data suggest that LGG alleviates gut inflammation, improves intestinal barrier function, and modulates the metabolite profile of piglets challenged with LPS. This trial was registered at the Zhejiang University (http://www.lac.zju.edu.cn) as ZJU20170529.
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http://dx.doi.org/10.1093/jn/nxaa009DOI Listing
May 2020

Identification of Metabonomics Changes in Longissimus Dorsi Muscle of Finishing Pigs Following Heat Stress through LC-MS/MS-Based Metabonomics Method.

Animals (Basel) 2020 Jan 13;10(1). Epub 2020 Jan 13.

State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

Heat stress (HS) negatively affects meat quality by affecting material and energy metabolism, and exploring the mechanism underlying the muscle response to chronic HS in finishing pigs is important for the global pork industry. This study investigated changes in the metabolic profiles of the longissimus dorsi (LD) muscle of finishing pigs under high temperature using ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) and multivariate data analysis (MDA). Castrated male DLY pigs (Duroc × Landrance × Yorkshire pigs, n = 24) from 8 litters were divided into three treatment groups: constant optimal ambient temperature at 22 °C and ad libitum feeding (CR, n = 8); constant high ambient temperature at 30 °C and ad libitum feeding (HS, n = 8); and constant optimal ambient temperature 22 °C and pair-feeding to the control pigs (PF, n = 8). The metabolic profile data from LD muscle samples were analyzed by MDA and external search engines. Nine differential metabolites (L-carnosine, acetylcholine, inosinic acid, L-carnitine, L-anserine, L-α-glycerylphosphorylcholine, acetylcarnitine, thiamine triphosphate, and adenosine thiamine diphosphate) were involved in antioxidant function, lipid metabolism, and cell signal transduction, which may decrease post mortem meat quality and play important roles in anti-HS. Four metabolites (L-carnosine, acetylcholine, inosinic acid, and L-carnitine) were verified, and it was indicated that the muscle L-carnitine content was significantly lower in HS than in CR ( < 0.01). The results show that constant HS affects the metabolites in the LD muscle and leads to coordinated changes in the endogenous antioxidant defense and meat quality of finishing pigs. These metabonomics results provide a basis for researching nutritional strategies to reduce the negative effects of heat stress on livestock and present new insights for further research.
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http://dx.doi.org/10.3390/ani10010129DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022765PMC
January 2020

miR-17-5p promotes proliferation and migration of CAL-27 human tongue squamous cell carcinoma cells involved in autophagy inhibition under hypoxia.

Int J Clin Exp Pathol 2019 1;12(6):2084-2091. Epub 2019 Jun 1.

Department of Oral and Maxillofacial Surgery, School of Stomatology, Shandong University Jinan, Shandong, China.

Autophagy contributes to head and neck squamous cell carcinoma (HNSCC) development and progression. MiR-17-5p down-regulates Beclin-1 and thus plays an important role in autophagy, but little is known about the function and regulation of miR-17-5p in HNSCC autophagy. This study aimed to investigate the role of miR-17-5p on proliferation, migration, and autophagy under hypoxia in CAL-27 human tongue squamous cell carcinoma cells. CAL-27 cells were transfected with 50 nmol miR-17-5p mimics to overexpress miR-17-5p. Cell proliferation and migration were determined by CCK-8 and wound healing assays, respectively, under hypoxia. Autophagy induced by hypoxia was detected by transmission electron microscope and Beclin-1 mRNA and protein expressions. The miR-17-5p mimics successfully increased the expression of miR-17-5p in CAL-27 cells by almost 700 fold compared with the miRNA mimic negative control. After 3 days, cells transfected with the miR-17-5p mimics showed higher proliferation compared with controls ( < 0.05) under hypoxia. MiR-17-5p transfected CAL-27 cells had a higher migratory capacity compared with the control cells ( < 0.05) under hypoxia. Furthermore, transmission electron microscopy showed that miR-17-5p overexpression inhibited the formation of autophagosomes in hypoxic cells. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot showed that miR-17-5p overexpression inhibited the mRNA and protein expression of Beclin-1 in CAL-27 cells submitted to hypoxia. MiR-17-5p overexpression promoted the proliferation and migration of the CAL-27 cells, but inhibited autophagy under hypoxia.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6949637PMC
June 2019

Poly-acrylic acid grafted natural rubber for multi-coated slow release compound fertilizer: Preparation, properties and slow-release characteristics.

Int J Biol Macromol 2020 Mar 7;146:540-548. Epub 2020 Jan 7.

Institute of Agricultural Resources Chemistry and Application, College of Science, Gansu Agricultural University, Lanzhou 730070, China.

In this study, a novel multi-coated slow release compound fertilizer based on natural rubber (NR) was prepared and characterized. Firstly, NR was grafted with poly-acrylic acid by in-situ radical solution polymerization to synthesize poly-acrylic acid grafted natural rubber (NR-g-PAA), the reaction conditions were optimized to increase the water absorption properties of NR-g-PAA. Through a series of characterization and test, the structure, morphology, thermal properties and biodegradability of NR-g-PAA were determined. Subsequently, a multi-nutrient fertilizer core was fabricated with urea, KHPO, and attapulgite by pan granulation. Then the fertilizer core was coated by NR as the inner layer and NR-g-PAA as the outer layer. Meanwhile, the slow release behavior of the compound fertilizer in soil was also studied. Results showed that the maximum water absorbency of NR-g-PAA is 744.00 ± 14.38%. The release rate of N, P and K in 30 days for NR/NR-g-PAA coated fertilizer was about 54.35 ± 1.49%, 51.18 ± 2.15% and 44.37 ± 1.38%, respectively, showing that the nutrient element release can last for >30 days. Overall, the novel method introduced in this study can inform the development of NR based controlled release fertilizers.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.01.051DOI Listing
March 2020

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Animals (Basel) 2019 Jun 28;9(7). Epub 2019 Jun 28.

College of Animal Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, Hangzhou 310058, China.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino acid, fatty acid, amines metabolism, and gut microbiome-derived metabolism. Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic acid were decreased, indicating insufficient source of fuel for energy supply, resulting in negative energy balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty acid (myristic acid, palmitic acid, and linoleic acid) and short-chain fatty acids (3-hydroxybutanoic acid and maleic acid) suggested fat decomposition compensating for energy shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic acid were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.
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http://dx.doi.org/10.3390/ani9070395DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6680871PMC
June 2019

Lactobacillus reuteri ZJ617 Culture Supernatant Attenuates Acute Liver Injury Induced in Mice by Lipopolysaccharide.

J Nutr 2019 11;149(11):2046-2055

College of Animal Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, Hangzhou, China.

Background: Lactobacillus rhamnosus GG culture supernatant (LGGs) promotes intestinal integrity and ameliorates acute liver injury induced by alcohol in mice.

Objectives: The aim of this study was to investigate the protective effects and molecular mechanisms of Lactobacillus reuteri ZJ617 culture supernatant (ZJ617s) on acute liver injury induced by lipopolysaccharide (LPS) in mice.

Methods: Male C57BL/6 mice (20 ± 2 g, 8 wk old) were randomly divided into 4 groups (6 mice/group): oral inoculation with phosphate-buffered saline (control), intraperitoneal injection of LPS (10 mg/kg body weight) (LPS), oral inoculation with ZJ617s 2 wk before intraperitoneal injection of LPS (ZJ617s + LPS), or oral inoculation with LGGs 2 wk before intraperitoneal injection of LPS (LGGs + LPS). Systemic inflammation, intestinal integrity, biomarkers of hepatic function, autophagy, and apoptosis signals in the liver were determined.

Results: Twenty-four hours after LPS injection, the activities of serum alanine transaminase and aspartate transaminase were 32.2% and 30.3% lower in the ZJ617s + LPS group compared with the LPS group, respectively (P < 0.05). The ZJ617s + LPS group exhibited higher intestinal expression of claudin 3 (62.5%), occludin (60.1%), and zonula occludens 1 (60.5%) compared with the LPS group (P < 0.05). The concentrations of hepatic interleukin-6 and tumor necrosis factor-α were 21.4% and 27.3% lower in the ZJ617s + LPS group compared with the LPS group, respectively (P < 0.05). However, the concentration of interleukin-10 was 22.2% higher in the ZJ617s + LPS group. LPS increased the expression of Toll-like receptor 4 (TLR4; by 50.5%), phosphorylation p38 mitogen-activated protein kinase (p38MAPK; by 57.1%), extracellular signal-regulated kinase (by 77.8%), c-Jun N-terminal kinase (by 42.9%), and nuclear factor-κB (NF-κB; by 36.0%) compared with the control group. Supplementation with ZJ617s or LGGs ameliorated these effects (P < 0.05). Moreover, the hepatic expression of active caspase-3 and microtubule-associated protein 1 light chain 3 II was 23.8% and 28.6% lower in the ZJ617s + LPS group compared with the LPS group, respectively (P < 0.05).

Conclusions: ZJ617s exerts beneficial effects on the mouse liver through suppression of hepatic TLR4/MAPK/NF-κB activation, apoptosis, and autophagy. This trial was registered at Zhejiang University (http://www.lac.zju.edu.cn) as NO.ZJU20170529.
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http://dx.doi.org/10.1093/jn/nxz088DOI Listing
November 2019

Salt-stress induced proteomic changes of two contrasting alfalfa cultivars during germination stage.

J Sci Food Agric 2019 Feb 25;99(3):1384-1396. Epub 2018 Oct 25.

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.

Background: Alfalfa (Medicago sativa L.), the primary forage crop throughout the world, is sensitive to salt stress during the germination stage. To investigate the response of alfalfa to salt stress, a comprehensive proteomic analysis was performed comparing alfalfa cultivars that differ in salinity tolerance in the early seedling.

Results: Five cultivars were examined for salt tolerance, and the most salt-tolerant cultivar, ZhongmuNo.3, and the most salt-sensitive cultivar, Daxiyang, were compared in terms of their physiological and proteomic responses. The two alfalfa cultivars seeds were exposed to 0 mmolL or 200 mmolL NaCl salt treatment for 10 days. Salt stress significantly reduced young seedling growth and the cotyledons' chlorophyll content; meanwhile, it increased the cotyledons' H O and malondialdehyde (MDA) levels, all of which were less adversely affected in ZhongmuNo.3 than in Daxiyang. A total of 51 spots (24 and 27 protein spots in the salt-sensitive and salt-tolerant cultivars, respectively) were identified as significantly differentially expressed using two-dimensional electrophoresis analysis. The proteins that were associated with salt tolerance included antioxidants/detoxifying enzymes, molecular chaperones, energy metabolic enzymes, a secondary metabolic enzyme, and pathogenesis-related proteins.

Conclusions: Under salt stress, ZhongmuNo.3 possessed a higher capacity for reactive oxygen species (ROS) scavenging, a more abundant energy supply, and stronger photosynthesis than the salt-sensitive cultivar Daxiyang, and these physiological processes may be the primary contributors to salt tolerance in ZhongmuNo.3. These advanced proteome data expand our knowledge of the physiology of the response of alfalfa to salt stress, providing a potentially valuable foundation for molecular breeding to enhance salt tolerance. © 2018 Society of Chemical Industry.
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http://dx.doi.org/10.1002/jsfa.9331DOI Listing
February 2019

Simvastatin Improves the Jaw Bone Microstructural Defect Induced by High Cholesterol Diet in Rats by Regulating Autophagic Flux.

Biomed Res Int 2018 27;2018:4147932. Epub 2018 Jun 27.

Department of Oral and Maxillofacial Surgery, Shandong Provincial Hospital, Affiliated to Shandong University, Jinan, Shandong 250012, China.

Objective: The objective of this study is to evaluate the effect of simvastatin on the jaw bone microstructural defect and autophagy in rats with high cholesterol diet (HCD).

Methods: Male Sprague-Dawley rats were fed a standard rodent chow (NC group) or a high cholesterol diet for 32 weeks and the HCD-fed rats were treated with vehicle (HC group) or simvastatin (5 mg/kg orally daily for 8 weeks, HC + SIM group, and = 10/group). The static histomorphometric changes in the jaw bone tissues in individual rats were evaluated. The relative levels of OPG, RANKL, NF-B, LC3, and p62 in the jaw bone tissues were determined by quantitative RT-PCR and/or immunohistochemistry.

Results: Compared with the NC group, the HC groups had lower trabecular bone volume, trabecular thickness and trabecular number, and increased ratios of RANKL/OPG in the jaw bone, accompanied by enhanced NF-B activation and autophagy. Simvastatin treatment inhabited these changes, including the decreased levels of serum proinflammatory cytokines and increased autophagy.

Conclusion: Simvastatin treatment could inhibit the hyperlipidemia-induced jaw bone microstructural defect in rats by increasing autophagic flux.
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http://dx.doi.org/10.1155/2018/4147932DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6040281PMC
January 2019

Genome-Wide Association and Functional Studies Identify and as Novel Susceptibility Genes for Coronary Artery Disease.

Arterioscler Thromb Vasc Biol 2018 04 22;38(4):964-975. Epub 2018 Feb 22.

From the Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, Beijing, China (Y.L., J.G., K.X., C.S., F.Y., B.Z., H.Y., W.-J.W., C.W., J.D., G.Z., X.-L.T.); Beijing Anzhen Hospital (Y.L.), Department of Cardiology, Beijing Anzhen Hospital (Y.Y., C.W.), and Department of Cardiology, Beijing Chaoyang Hospital (F.J.), Capital Medical University, China; Beijing Institute of Heart, Lung, and Blood Vessel Disease, China (Y.L.); Department of Internal Medicine, Gene Therapy Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (D.W.W., H.D., S.T.); Department of Cardiology, The General Hospital of Chinese People's Liberation Army, Beijing, China (Y.C., Z.S.); Cardiovascular Medicine Center, Affiliated Hospital of Guangdong Medical University, Zhanjiang, China (C.C., S.H., J.Z., Y.M., W.C., J.S., W.L.); State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing (S.Z., X.L., X.Y., D.G.); Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, China (L.Z.); Department of Internal Medicine, Hospital of Peking University, Beijing, China (Y.C.); Department of Cardiology, The Second Affiliated Hospital of Harbin Medical University, China (L.J.); Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, China (G.L.); Cardiovascular Department, Ningbo First Hospital, Ningbo University, China (H.C., S.W.); Institut für Integrative und Experimentelle Genomik, Universität zu Lübeck, Germany (J.E.); German Research Centre for Cardiovascular Research, Partner Site Hamburg/Lübeck/Kiel (J.E.); Deutsches Herzzentrum München, Technische Universität München, Germany (L.Z., H.S.); German Centre for Cardiovascular Research, Partner Site Munich Heart Alliance (L.Z., H.S.); Department of Human Population Genetics, Human Aging Research Institute and School of Life Science, Nanchang University, China (S.C., X.-L.T.); and Institute of Medical Systems Biology (S.R.) and School of Public Health (S.R.), Guangdong Medical University, Dongguan, China.

Objective: The genetic contribution to coronary artery disease (CAD) remains largely unclear. We combined genetic screening with functional characterizations to identify novel loci and candidate genes for CAD.

Approach And Results: We performed genome-wide screening followed by multicenter validation in 8 cohorts consisting of 21 828 participants of Han ethnicity and identified 3 novel intragenic SNPs (single nucleotide polymorphisms), rs9486729 ( [Scm polycomb group protein-like 4]; odds ratio, 1.25; 95% CI, 1.17-1.34; =3.51×10), rs17165136 ( [thrombospondin type 1 domain-containing 7A]; odds ratio 1.28; 95% CI, 1.21-1.35; <1.00×10), and rs852787 ( [disabled-1]; odds ratio, 1.29; 95% CI, 1.21-1.38; =2.02×10), associated with CAD with genome-wide significance. The risk allele of rs9486729 and protective allele of rs17165136 were associated with the decreased expression of their host genes, and , respectively, whereas rs852787 did not have transcriptional effects on any gene. Knockdown of activated endothelial cells by increasing the expression of , , and and weakened their antiapoptotic activity, whereas the knockdown of had little effect on these endothelial cell functions but attenuated monocyte adhesion via decreasing the expression of , , and . We further showed that inhibiting the expression of exacerbated endothelial dysfunction and vascular remodeling in a rat model with partial carotid ligation.

Conclusions: We identify 3 novel loci associated with CAD and show that 2 genes, and , make functional contributions to atherosclerosis. How rs852787 and its host gene are linked to CAD needs further studies.
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http://dx.doi.org/10.1161/ATVBAHA.117.310594DOI Listing
April 2018

Proteomic changes of the porcine skeletal muscle in response to chronic heat stress.

J Sci Food Agric 2018 Jul 27;98(9):3315-3323. Epub 2018 Feb 27.

State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.

Background: Heat stress (HS) has an adverse effect on meat quality; however, the underlying molecular mechanisms altering meat quality due to muscle responses to stress remain unclear. Sixteen castrated male crossbreeds between Landrace × Yorkshire sows and Duroc boars (79.00 ± 1.50 kg body weight) were exposed to either thermal neutral (22 °C, n = 8) or HS (30 °C, n = 8) conditions for 3 weeks. Subsequently, the longissimus dorsi (LD) muscle of all pigs was assayed for meat quality parameters and proteome analysis.

Results: HS decreased post mortem (24 h) pH and intramuscular fat, changed ultimate L*, a* and b* values and increased drip loss and shear force. Proteome analysis of the LD was conducted by two-dimensional gel electrophoresis and mass spectrometry. A total of 23 differentially expressed proteins were identified, of which three were verified by western blotting analysis. The identified proteins were involved in six types of biological process: carbohydrate metabolism, myofibrillar and cytoskeleton structure, stress response, antioxidant and detoxification, calcium binding and cellular apoptosis. Interestingly, HS induced higher levels of heat shock protein, antioxidants and calcium binding proteins, which are involved in the mechanisms of defense and homeostasis.

Conclusion: The results indicate that HS-induced changes in the expression of myofibrillar proteins, glucose and energy metabolism-related proteins, heat shock protein and antioxidant enzymes might, at least partly, contribute to increase in meat tenderness. These findings will provide the foundation for developing future mitigating solutions and preventative therapies to reduce the detrimental effects of chronic HS on muscle function, metabolism and meat quality. © 2017 Society of Chemical Industry.
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http://dx.doi.org/10.1002/jsfa.8835DOI Listing
July 2018

ZJ617 maintains intestinal integrity via regulating tight junction, autophagy and apoptosis in mice challenged with lipopolysaccharide.

Oncotarget 2017 Sep 24;8(44):77489-77499. Epub 2017 Aug 24.

Institute of Animal Nutrition, College of Animal Science and Technology, Zhejiang A & F University, Lin'an 311300, P.R. China.

Live probiotics are effective in reducing gut permeability and inflammation. We have previously reported that ZJ617 (ZJ617) with high adhesive and GG (LGG) can ameliorate intestine inflammation induced by lipopolysaccharide (LPS). The present study was aimed at elucidating the roles of ZJ617 and LGG in alleviating the LPS-induced barrier dysfunction of ileum in mice. Six C57BL/6 mice per group were orally inoculated with ZJ617 or LGG for one week (1× 10 CFU/mouse) and intraperitoneally injected with LPS (10 mg/kg body weight) for 24 h. The results demonstrated that pretreatment with ZJ617 and LGG attenuated LPS-induced increase in intestinal permeability. The probiotics supplementation suppressed LPS-induced oxidative stress. Both ZJ617 and LGG strongly reversed the decline of occludin and claudin-3 expression induced by LPS challenge. ZJ617 relieved LPS-induced apoptosis by decreasing caspase-3 activity. Noticeably, ratio of microtubule-associated light chain 3 (LC3)-II/LC3-I and LC3 activity were elevated by LPS stimulation, whereas such increases were obviously attenuated by both of the probiotics treatment. Moreover, phosphorylated mammalian target of rapamycin (p-mTOR) was significantly inhibited by LPS, whereas complementation of ZJ617 and LGG markedly increased the expression of p-mTOR. Collectively, our results indicated that ZJ617 could protect LPS-induced intestinal barrier dysfunction via enhancing antioxidant activities and tight junction and attenuating apoptosis and autophagy via mTOR signaling pathway. These findings could serve as systematic mechanisms through which probiotics promote and maintain gut homeostasis.
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http://dx.doi.org/10.18632/oncotarget.20536DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5652795PMC
September 2017

Influence of tetramethylenedisulfotetramine on synchronous calcium oscillations at distinct developmental stages of hippocampal neuronal cultures.

Neurotoxicology 2017 01 29;58:11-22. Epub 2016 Oct 29.

Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, 95616, United States. Electronic address:

The spatial and temporal patterns of spontaneous synchronous Ca oscillations (SCOs) regulate physiological pathways that influence neuronal development, excitability, and health. Hippocampal neuronal cultures (HN) and neuron/glia co-cultures (HNG) produced from neonatal mice were loaded with Fluo-4/AM and SCOs recorded in real-time using a Fluorescence Imaging Plate Reader at different developmental stages in vitro. HNG showed an earlier onset of SCOs, with low amplitude and low frequency SCOs at 4days in vitro (DIV), whereas HN were quiescent at this point. SCO amplitude peaked at 9 DIV for both cultures. SCO network frequency peaked at 12 DIV in HN, whereas in HNG the frequency peaked at 6 DIV. SCO patterns were associated with the temporal development of neuronal networks and their ratio of glutamatergic to GABAergic markers of excitatory/inhibitory balance. HN and HNG exhibited differential responses to the convulsant tetramethylenedisulfotetramine (TETS) and were highly dependent on DIV. In HN, TETS triggered an acute rise of intracellular Ca (Phase I response) only in 14 DIV and a sustained decrease of SCO frequency with increased amplitude (Phase II response) at all developmental stages. In HNG, TETS decreased the SCO frequency and increased the amplitude at 6 and 14 but not 9 DIV. There was no acute Ca rise (Phase I response) in any age of HNG tested with TETS. These data demonstrated the importance of glia and developmental stage in modulating neuronal responses to TETS. Our results illustrate the applicability of the model for investigating how caged convulsants elicit abnormal network activity during the development of HN and HNG cultures in vitro.
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http://dx.doi.org/10.1016/j.neuro.2016.10.015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5303550PMC
January 2017

Transcriptome analysis reveals that constant heat stress modifies the metabolism and structure of the porcine longissimus dorsi skeletal muscle.

Mol Genet Genomics 2016 Dec 25;291(6):2101-2115. Epub 2016 Aug 25.

State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, China.

Exposure to high ambient temperatures is detrimental to pig rearing and porcine meat quality. Deep molecular sequencing allows for genomic characterization of porcine skeletal muscles and helps understand how the genomic landscape may impact meat quality. To this end, we performed mRNA-seq to molecularly dissect the impact of heat stress on porcine skeletal muscles, longissimus dorsi. Sixteen castrated, male DLY pigs [which are crossbreeds between Duroc (D) boars and Landrace (L) × Yorkshire (Y) sows, 79.0 ± 1.5 kg BW] were evenly split into two groups that were subjected to either control (CON) (22 °C; 55 % humidity) or constant heat stress (H30; 30 °C; 55 % humidity) conditions for 21 days. Seventy-eight genes were found to be differentially expressed, of which 37 were up-regulated and 41 were down-regulated owing to constant heat stress. We predicted 5247 unknown genes and 6108 novel transcribed units attributed to alternative splicing (AS) events in the skeletal muscle. Furthermore, 30,761 and 31,360 AS events were observed in the CON and H30 RNA-seq libraries, respectively. The differentially expressed genes in the porcine skeletal muscles were involved in glycolysis, lactate metabolism, lipid metabolism, cellular defense, and stress responses. Additionally, the expression levels of these genes were associated with variations in meat quality between the CON and H30 groups, indicating that heat stress modulated genes crucial to skeletal muscle development and metabolism. Our transcriptomic analysis provides valuable information for understanding the molecular mechanisms governing porcine skeletal muscle development. Such insights may lead to innovative strategies to improve meat quality of pigs under heat stress.
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http://dx.doi.org/10.1007/s00438-016-1242-8DOI Listing
December 2016

Genome-wide DNA methylation profiles changes associated with constant heat stress in pigs as measured by bisulfite sequencing.

Sci Rep 2016 06 6;6:27507. Epub 2016 Jun 6.

State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China.

Heat stress affects muscle development and meat quality in food animals; however, little is known regarding its regulatory mechanisms at the epigenetic level, such as via DNA methylation. In this study, we aimed to compare the DNA methylation profiles between control and heat-stressed pigs to identify candidate genes for skeletal muscle development and meat quality. Whole-genome bisulfite sequencing was used to investigate the genome-wide DNA methylation patterns in the longissimus dorsi muscles of the pigs. Both groups showed similar proportions of methylation at CpG sites but exhibited different proportions at non-CpG sites. A total of 57,147 differentially methylated regions were identified between the two groups, which corresponded to 1,422 differentially methylated genes. Gene ontogeny and KEGG pathway analyses indicated that these were mainly involved in energy and lipid metabolism, cellular defense and stress responses, and calcium signaling pathways. This study revealed the global DNA methylation pattern of pig muscle between normal and heat stress conditions. The result of this study might contribute to a better understanding of epigenetic regulation in pig muscle development and meat quality.
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http://dx.doi.org/10.1038/srep27507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4893741PMC
June 2016

Chronic Heat Stress Induces Immune Response, Oxidative Stress Response, and Apoptosis of Finishing Pig Liver: A Proteomic Approach.

Int J Mol Sci 2016 May 11;17(5). Epub 2016 May 11.

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Beijing 100193, China.

Heat stress (HS) negatively affects human health, animal welfare, and livestock production. We analyzed the hepatic proteomes of finishing pigs subjected to chronic heat stress (HS), thermal neutral (TN), and restricted feed intake conditions, identifying differences between direct and indirect (via reduced feed intake) HS. Twenty-four castrated male pigs were randomly allocated to three treatments for three weeks: (1) thermal neutral (TN) (22 °C) with ad libitum feeding; (2) chronic HS (30 °C) with ad libitum feeding; and (3) TN, pair-fed to HS intake (PF). Hepatic proteome analysis was conducted using two-dimensional gel electrophoresis and mass spectrometry. Both HS and PF significantly reduced liver weight (p < 0.05). Forty-five hepatic proteins were differentially abundant when comparing HS with TN (37), PF with TN (29), and HS with PF (16). These proteins are involved in heat shock response and immune defense, oxidative stress response, cellular apoptosis, metabolism, signal transduction, and cytoskeleton. We also observed increased abundance of proteins and enzymes associated with heat shock response and immune defense, reduced the redox state, enhanced multiple antioxidant abilities, and increased apoptosis in HS liver. Heat-load, independent of reduced feed intake, induced an innate immune response, while food restriction caused stress and cellular apoptosis. Our results provide novel insights into the effects of chronic HS on liver.
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http://dx.doi.org/10.3390/ijms17050393DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4881434PMC
May 2016

Proteomic changes of the porcine small intestine in response to chronic heat stress.

J Mol Endocrinol 2015 Dec 28;55(3):277-93. Epub 2015 Sep 28.

State Key Laboratory of Animal NutritionInstitute of Animal Sciences, Chinese Academy of Agricultural Science, Beijing 100193, People's Republic of China

Acute heat stress (HS) negatively affects intestinal integrity and barrier function. In contrast, chronic mild HS poses a distinct challenge to animals. Therefore, this study integrates biochemical, histological and proteomic approaches to investigate the effects of chronic HS on the intestine in finishing pigs. Castrated male crossbreeds (79.00 ± 1.50 kg BW) were subjected to either thermal neutral (TN, 21 °C; 55% ± 5% humidity; n=8) or HS conditions (30 °C; 55% ± 5% humidity; n=8) for 3 weeks. The pigs were sacrificed after 3 weeks of high environmental exposure and the plasma hormones, the intestinal morphology, integrity, and protein profiles of the jejunum mucosa were determined. Chronic HS reduced the free triiodothyronine (FT3) and GH levels. HS damaged intestinal morphology, increased plasma d-lactate concentrations and decreased alkaline phosphatase activity of intestinal mucosa. Proteome analysis of the jejunum mucosa was conducted by 2D gel electrophoresis and mass spectrometry. Fifty-three intestinal proteins were found to be differentially abundant, 18 of which were related to cell structure and motility, and their changes in abundance could comprise intestinal integrity and function. The down-regulation of proteins involved in tricarboxylic acid cycle (TCA cycle), electron transport chain (ETC), and oxidative phosphorylation suggested that chronic HS impaired energy metabolism and thus induced oxidative stress. Moreover, the changes of ten proteins in abundance related to stress response and defense indicated pigs mediated long-term heat exposure and counteracted its negative effects of heat exposure. These findings have important implications for understanding the effect of chronic HS on intestines.
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http://dx.doi.org/10.1530/JME-15-0161DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4632496PMC
December 2015

Rapid throughput analysis demonstrates that chemicals with distinct seizurogenic mechanisms differentially alter Ca2+ dynamics in networks formed by hippocampal neurons in culture.

Mol Pharmacol 2015 Apr 12;87(4):595-605. Epub 2015 Jan 12.

State Key Laboratory of Natural Medicines and Jiangsu Provincial Key laboratory for TCM Evaluation and Translational Development, China Pharmaceutical University, Nanjing, P.R. China (Z.C., X.Z., Y.C.); Department of Molecular Biosciences, School of Veterinary Medicine (Z.C., Y.C., S.H., P.J.L., I.N.P.) and Department of Pharmacology, School of Medicine (H.W.),University of California, Davis, California

Primary cultured hippocampal neurons (HN) form functional networks displaying synchronous Ca(2+) oscillations (SCOs) whose patterns influence plasticity. Whether chemicals with distinct seizurogenic mechanisms differentially alter SCO patterns was investigated using mouse HN loaded with the Ca(2+) indicator fluo-4-AM. Intracellular Ca(2+) dynamics were recorded from 96 wells simultaneously in real-time using fluorescent imaging plate reader. Although quiescent at 4 days in vitro (DIV), HN acquired distinctive SCO patterns as they matured to form extensive dendritic networks by 16 DIV. Challenge with kainate, a kainate receptor (KAR) agonist, 4-aminopyridine (4-AP), a K(+) channel blocker, or pilocarpine, a muscarinic acetylcholine receptor agonist, caused distinct changes in SCO dynamics. Kainate at <1 µM produced a rapid rise in baseline Ca(2+) (Phase I response) associated with high-frequency and low-amplitude SCOs (Phase II response), whereas SCOs were completely repressed with >1 µM kainate. KAR competitive antagonist CNQX [6-cyano-7-nitroquinoxaline-2,3-dione] (1-10 µM) normalized Ca(2+) dynamics to the prekainate pattern. Pilocarpine lacked Phase I activity but caused a sevenfold prolongation of Phase II SCOs without altering either their frequency or amplitude, an effect normalized by atropine (0.3-1 µM). 4-AP (1-30 µM) elicited a delayed Phase I response associated with persistent high-frequency, low-amplitude SCOs, and these disturbances were mitigated by pretreatment with the KCa activator SKA-31 [naphtho[1,2-d]thiazol-2-ylamine]. Consistent with its antiepileptic and neuroprotective activities, nonselective voltage-gated Na(+) and Ca(2+) channel blocker lamotrigine partially resolved kainate- and pilocarpine-induced Ca(2+) dysregulation. This rapid throughput approach can discriminate among distinct seizurogenic mechanisms that alter Ca(2+) dynamics in neuronal networks and may be useful in screening antiepileptic drug candidates.
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http://dx.doi.org/10.1124/mol.114.096701DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4366799PMC
April 2015

Mandelic acid chiral separation utilizing a two-phase partitioning bioreactor built by polysulfone microspheres and immobilized enzymes.

Bioprocess Biosyst Eng 2015 Mar 12;38(3):429-35. Epub 2014 Sep 12.

State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, Institute of Biochemical Engineering and Environmental Technology, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China,

A novel two-phase partitioning bioreactor (TPPB) modified by polysulfone (PSF) microspheres and immobilized enzyme (novozym-435) was formed, and the resulting TPPB was applied into mandelic acid chiral separation. The PSF microspheres containing n-hexanol (named PSF/hexanol microspheres) was prepared by using the phase inversion method, which was used as the organic phase. Meanwhile, the immobilized enzyme novozym-435 was used as a biocatalyst. The water phase was composed of the phosphate buffer solution (PBS). (R, S)-Methyl mandelate was selected as the substrate to study enzymatic properties. Different reaction factors have been researched, such as pH, reaction time, temperature and the quantity of biocatalyst and PSF/hexanol microspheres added in. Finally, (S)-mandelic acid was obtained with an 80 % optical purity after 24 h in the two-phase partitioning bioreactor. The enantiomeric excess (eep) values were very low in the water phase, in which the highest eep value was only 46 %. The eep of the two-phase partitioning bioreactor had been enhanced more obviously than that catalyzed in the water phase.
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http://dx.doi.org/10.1007/s00449-014-1283-5DOI Listing
March 2015

Gambierol inhibition of voltage-gated potassium channels augments spontaneous Ca2+ oscillations in cerebrocortical neurons.

J Pharmacol Exp Ther 2014 Sep 23;350(3):615-23. Epub 2014 Jun 23.

State Key Laboratory of Natural Medicines, Department of Complex Prescription of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, People's Republic of China (Z.C.); Department of Pharmacology, School of Medicine, Creighton University, Omaha, Nebraska (Z.C., Y.C., E.B., S.M., T.F.M.); and Department of Chemistry, University of Utah, Salt Lake City, Utah (J.D.R.)

Gambierol is a marine polycyclic ether toxin produced by the marine dinoflagellate Gambierdiscus toxicus and is a member of the ciguatoxin toxin family. Gambierol has been demonstrated to be either a low-efficacy partial agonist/antagonist of voltage-gated sodium channels or a potent blocker of voltage-gated potassium channels (Kvs). Here we examined the influence of gambierol on intact cerebrocortical neurons. We found that gambierol produced both a concentration-dependent augmentation of spontaneous Ca(2+) oscillations, and an inhibition of Kv channel function with similar potencies. In addition, an array of selective as well as universal Kv channel inhibitors mimicked gambierol in augmenting spontaneous Ca(2+) oscillations in cerebrocortical neurons. These data are consistent with a gambierol blockade of Kv channels underlying the observed increase in spontaneous Ca(2+) oscillation frequency. We also found that gambierol produced a robust stimulation of phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2). Gambierol-stimulated ERK1/2 activation was dependent on both inotropic [N-methyl-d-aspartate (NMDA)] and type I metabotropic glutamate receptors (mGluRs) inasmuch as MK-801 [NMDA receptor inhibitor; (5S,10R)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate], S-(4)-CGP [S-(4)-carboxyphenylglycine], and MTEP [type I mGluR inhibitors; 3-((2-methyl-4-thiazolyl)ethynyl) pyridine] attenuated the response. In addition, 2-aminoethoxydiphenylborane, an inositol 1,4,5-trisphosphate receptor inhibitor, and U73122 (1-[6-[[(17b)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione), a phospholipase C inhibitor, both suppressed gambierol-induced ERK1/2 activation, further confirming the role of type I mGluR-mediated signaling in the observed ERK1/2 activation. Finally, we found that gambierol produced a concentration-dependent stimulation of neurite outgrowth that was mimicked by 4-aminopyridine, a universal potassium channel inhibitor. Considered together, these data demonstrate that gambierol alters both Ca(2+) signaling and neurite outgrowth in cerebrocortical neurons as a consequence of blockade of Kv channels.
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http://dx.doi.org/10.1124/jpet.114.215319DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4152883PMC
September 2014

Nanomolar bifenthrin alters synchronous Ca2+ oscillations and cortical neuron development independent of sodium channel activity.

Mol Pharmacol 2014 Apr 30;85(4):630-9. Epub 2014 Jan 30.

Department of Molecular Biosciences, School of Veterinary Medicine (Z.C., Y.C., I.N.P.), and Department of Pharmacology, School of Medicine (H.M.N., D.P.J., H.W.), University of California Davis, Davis, California.

Bifenthrin, a relatively stable type I pyrethroid that causes tremors and impairs motor activity in rodents, is broadly used. We investigated whether nanomolar bifenthrin alters synchronous Ca(2+) oscillations (SCOs) necessary for activity-dependent dendritic development. Primary mouse cortical neurons were cultured 8 or 9 days in vitro (DIV), loaded with the Ca(2+) indicator Fluo-4, and imaged using a Fluorescence Imaging Plate Reader Tetra. Acute exposure to bifenthrin rapidly increased the frequency of SCOs by 2.7-fold (EC50 = 58 nM) and decreased SCO amplitude by 36%. Changes in SCO properties were independent of modifications in voltage-gated sodium channels since 100 nM bifenthrin had no effect on the whole-cell Na(+) current, nor did it influence neuronal resting membrane potential. The L-type Ca(2+) channel blocker nifedipine failed to ameliorate bifenthrin-triggered SCO activity. By contrast, the metabotropic glutamate receptor (mGluR)5 antagonist MPEP [2-methyl-6-(phenylethynyl)pyridine] normalized bifenthrin-triggered increase in SCO frequency without altering baseline SCO activity, indicating that bifenthrin amplifies mGluR5 signaling independent of Na(+) channel modification. Competitive [AP-5; (-)-2-amino-5-phosphonopentanoic acid] and noncompetitive (dizocilpine, or MK-801 [(5S,10R)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate]) N-methyl-d-aspartate antagonists partially decreased both basal and bifenthrin-triggered SCO frequency increase. Bifenthrin-modified SCO rapidly enhanced the phosphorylation of cAMP response element-binding protein (CREB). Subacute (48 hours) exposure to bifenthrin commencing 2 DIV-enhanced neurite outgrowth and persistently increased SCO frequency and reduced SCO amplitude. Bifenthrin-stimulated neurite outgrowth and CREB phosphorylation were dependent on mGluR5 activity since MPEP normalized both responses. Collectively these data identify a new mechanism by which bifenthrin potently alters Ca(2+) dynamics and Ca(2+)-dependent signaling in cortical neurons that have long term impacts on activity driven neuronal plasticity.
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http://dx.doi.org/10.1124/mol.113.090076DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3965893PMC
April 2014

The performance analysis of distributed Brillouin corrosion sensors for steel reinforced concrete structures.

Sensors (Basel) 2013 Dec 27;14(1):431-42. Epub 2013 Dec 27.

College of Physics and Optoelectronic Engineering, Dalian University of Technology, Dalian 116024, China.

The Brillouin optical time-domain analysis (BOTDA)-based optical fiber method has been proposed to measure strain variations caused by corrosion expansion. Spatial resolutions of 1 m can be achieved with this kind of Brillouin sensor for detecting the distributed strain. However, when the sensing fiber is wound around the steel rebar in a number of circles in a range of several meters, this spatial resolution still has limitations for corrosion monitoring. Here, we employed a low-coherent fiber-optic strain sensor (LCFS) to survey the performance of Brillouin sensors based on the fact that the deformation measured by the LCFS equals the integral of the strains obtained from Brillouin sensors. An electrochemical accelerated corrosion experiment was carried out and the corrosion expansion was monitored by both BOTDA and the LCFS. Results demonstrated that the BOTDA can only measure the expansion strain of about 1,000 με, which was generated by the 18 mm steel rebar corrosion, but, the LCFS had high sensitivity from the beginning of corrosion to the destruction of the structure, and no obvious difference in expansion speed was observed during the acceleration stage of the corrosion developed in the reinforced concrete (RC) specimens. These results proved that the BOTDA method could only be employed to monitor the corrosion inside the structure in the early stage.
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http://dx.doi.org/10.3390/s140100431DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3926566PMC
December 2013

Superparamagnetic polymer emulsion particles from a soap-free seeded emulsion polymerization and their application for lipase immobilization.

Appl Biochem Biotechnol 2014 Jan 10;172(2):701-12. Epub 2013 Oct 10.

State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China.

Using emulsion copolymer of styrene (St), glycidyl methacrylate (GMA) and 2-hydroxyethyl methacrylate (HEMA) as seed latexes, the superparamagnetic polymer emulsion particles were prepared by seeded emulsion copolymerization of butyl methacrylate (BMA), vinyl acetate (VAc) and ethylene glycol dimethacrylate in the presence of the seed latexes and superparamagnetic Fe3O4/SiOx nanoparticles (or Fe3O4-APTS nanoparticles) through a two-step process, without addition of any emulsifier. The magnetic emulsion particles named P(St-GMA-HEMA)/P(BMA-VAc) were characterized by transmission electron microscope and vibrating sample magnetometry. The results showed that the magnetic emulsion particles held a structure with a thinner shell (around 100 nm) and a bigger cavity (around 200 nm), and possessed a certain level of magnetic response. The resulting magnetic emulsion particles were employed in the immobilization of lipase by two strategies to immobilized lipase onto the resulting magnetic composites directly (S-1) or using glutaraldehyde as a coupling agent (S-2), thus, experimental data showed that the thermal stability and reusability of immobilized lipase based on S-2 were higher than that of S-1.
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http://dx.doi.org/10.1007/s12010-013-0563-xDOI Listing
January 2014
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