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Having enterprises engaged in environmentally friendly behavior is an important part of reducing negative environmental impacts. This study makes a quantitative analysis against the backdrop of China's transitional economic system. The results show that politically-connected enterprises significantly reduce environmental expenditure, but this only holds for state-owned enterprises; private enterprises with political connections spend significantly more. Analysis of the efficiency of environmental expenditure indicates that, for private enterprises, environmental spending is used as a way to maintain political connections, with rent-seeking as the likely motivation. Politically-connected private enterprises have not reduced their emissions to the same extent as state-owned enterprises, despite increased expenditure. Given the scale of environmental degradation in China during a period of massive economic and social upheaval, the results of this analysis provide a quantitative case for policy change: governments should shift focus to the results that environmental spending produces.
In the current research, a 60-day experiment was conducted with the purpose of exploring the impacts of methionine on growth performance, muscle nutritive deposition, muscle fibre growth and type I collagen synthesis as well as related signaling pathway. Six diets (iso-nitrogenous) differing in methionine concentrations (2.54, 4.85, 7.43, 10.12, 12.40 and 15.11 g kg-1 diets) were fed to 540 grass carp (178.47 ± 0.36 g). Results showed (P < 0.05) that, compared with methionine deficiency, optimal level of dietary methionine (1) increased feed intake (FI), feed efficiency (FE), specific growth rate (SGR) and percentage weight gain (PWG); (2) increased fish muscle protein, lipid and free amino acid contents, and improved fish muscle fatty acid profile as well as increased protein content in part associated with TORC1/S6K1 signaling pathway; (3) increased the frequency distribution of muscle fibre with >50 µm of diameter; (4) increased type I collagen synthesis partly related to TGF-β1/Smads and CK2/TORC1 signaling pathways. In conclusion, dietary methionine improved muscle growth, which might be due to the regulation of muscle nutritive deposition, and muscle fibre growth and type I collagen synthesis related signal molecules. Finally, according to PWG and muscle collagen content, the methionine requirements for on-growing grass carp (178-626 g) were estimated to be 9.56 g kg-1 diet (33.26 g kg-1 protein of diet) and 9.28 g kg-1 diet (32.29 g kg-1 of dietary protein), respectively.
Many emotional experiences such as anxiety and depression are influenced by negative affect (NA). NA has both trait and state features, which play different roles in physiological and mental health. Attending to NA common to various emotional experiences and their trait-state features might help deepen the understanding of the shared foundation of related emotional disorders.
The principal component of five measures was calculated to indicate individuals' NA level. Applying the connectivity-based correlation analysis, we first identified resting-state functional connectives (FCs) relating to NA in sample 1 (n = 367), which were validated through an independent sample (n = 232; sample 2). Next, based on the variability of FCs across large timescale, we further divided the NA-related FCs into high- and low-variability groups. Finally, FCs in different variability groups were separately applied to predict individuals' neuroticism level (which is assumed to be the core trait-related factor underlying NA), and the change of NA level (which represents the state-related fluctuation of NA).
The low-variability FCs were primarily within the default mode network (DMN) and between the DMN and dorsal attention network/sensory system and significantly predicted trait rather than state NA. The high-variability FCs were primarily between the DMN and ventral attention network, the fronto-parietal network and DMN/sensory system, and significantly predicted the change of NA level.
The trait and state NA can be separately predicted by stable and variable spontaneous FCs with different attentional processes and emotion regulatory mechanisms, which could deepen our understanding of NA.
The experiment was conducted to investigate the effects of dietary threonine (Thr) on growth performance and muscle growth, protein synthesis and antioxidant-related signalling pathways of hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂. A total of 1200 fish (14·19 (se 0·13) g) were randomly distributed into six groups with four replicates each, fed six diets with graded level of Thr (9·5, 11·5, 13·5, 15·4, 17·4 and 19·3 g/kg diets) for 56 d. Results showed (P < 0·05) that dietary Thr (1) increased percentage weight gain, specific growth rate, feed efficiency and protein efficiency ratio; (2) up-regulated growth hormone (GH), insulin-like growth factor 1 (IGF-1), proliferating cell nuclear antigen, myogenic regulation factors (MyoD, Myf5, MyoG and Mrf4) and myosin heavy chain (MyHC) mRNA levels; (3) increased muscle protein content via regulating the protein kinase B/target of rapamycin signalling pathway and (4) decreased malondialdehyde and protein carbonyl contents, increased catalase, glutathione-S-transferase, glutathione reductase and GSH activities, up-regulated mRNA levels of antioxidant enzymes related to NFE2-related factor 2 and γ-glutamylcysteine ligase catalytic subunit. These results suggest that Thr has a potential role to improve muscle growth and protein synthesis, which might be due to the regulation of GH-IGF system, muscle growth-related gene, antioxidative capacity and protein synthesis-related signalling pathways. Based on the quadratic regression analysis of specific growth rate, the Thr requirement of hybrid catfish (14·19–25·77 g) was estimated to be 13·77 g/kg of the diet (33·40 g/kg of dietary protein).
The present study investigated the effects of condensed tannins (CT) on intestinal immune function in on-growing grass carp (Ctenopharyngodon idella). A total of 540 healthy grass carp were fed six diets containing different levels of CT (0, 10·00, 20·00, 30·00, 40·00 and 50·00 g/kg diet) for 70 d and then challenged with Aeromonas hydrophila for 14 d. The results showed that, compared with the control group, dietary CT (1) induced intestinal histopathological lesions and aggravated enteritis; (2) decreased lysozyme and acid phosphatase activities, complement 3 (C3), C4 and IgM contents and down-regulated the Hepcidin, liver-expressed antimicrobial peptide (LEAP)-2A, LEAP-2B, Mucin2 and β-defensin-1 mRNA levels in the proximal intestine (PI), mid intestine (MI) and distal intestine (DI) (P < 0·05); (3) down-regulated the mRNA levels of anti-inflammatory cytokines transforming growth factor (TGF)-β1, TGF-β2 (not in MI and DI), IL-4/13A (not IL-4/13B), IL-10 and IL-11 partly correlated with target of rapamycin (TOR) signalling; and (4) up-regulated the mRNA levels of pro-inflammatory cytokines interferon-γ2, IL-1β, IL-6, IL-8 (not in PI), IL-12p35, IL-12p40, IL-15 and IL-17D partly related to NF-κB signalling in the intestine of on-growing grass carp. Overall, the results indicated that CT could impair the intestinal immune function, and its potential regulation mechanisms were partly associated with the TOR and NF-κB signalling pathways. Finally, based on the percentage weight gain and enteritis morbidity, the maximum allowable levels of CT for on-growing grass carp (232·22–890·11 g) were estimated to be 18·6 and 17·4 g/kg diet, respectively.
Imaging studies have shown that the subcallosal region (SCR) volume was decreased in patients with major depressive disorder (MDD). However, whether the volumetric reductions in the SCR are due to thinning of the cortex or a loss of surface area (SA) remains unclear. In addition, the relationship between cortical measurements of the SCR and age through the adult life span in MDD remains unclear.
We used a cross-sectional design from 114 individuals with MDD and 112 matched healthy control (HC) individuals across the adult life span (range: 18–74 years). The mean cortical volume (CV), SA and cortical thickness (CT) of the SCR were computed using cortical parcellation based on FreeSurfer software. Multivariate analyses of covariance models were performed to compare differences between the MDD and HC groups on cortical measurements of the SCR. Multiple linear regression models were used to test age-by-group interaction effects on these cortical measurements of the SCR.
The MDD had significant reductions in the CV and SA of the left SCR compared with HC individuals after controlling of other variables. The left SCR CV and SA reductions compared with matched controls were observed only in early adulthood patients. We also found a significant age-related CT reduction in the SCR both in the MDD and HC participants.
The SCR volume reduction was mainly driven by SA in MDD. The different trajectories between the CT and SA of the SCR with age may provide valuable information to distinguish pathological processes and normal ageing in MDD.
Chitooligosaccharides (COS) are multi-functional foods and nutrients and environmentally friendly biological abiotic-resistance inducing agents for plants. In the current study, the effects and possible mechanisms of COS on improving the cold resistance of rice (II YOU 1259) seedlings were investigated. Compared with the control, a COS pre-soaking treatment enhanced photosynthesis, reduced oxidation damage and led to accumulation of more osmotic regulation substances under chilling treatment. In addition, a novel Deg/HtrA family serine endopeptidase (DegQ) gene, related to COS enhanced rice cold resistance, was identified. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that transcription of DegQ and psbA (D1 protein encoding gene) were up-regulated in a time-dependent manner by COS treatment under cold stress. With increasing expression of the D1 protein, chlorophyll b content was enhanced correspondingly. The current results suggest that COS could enhance cold stress tolerance of rice by repairing the photodamaged photosystem II, altering osmotic regulation and reducing oxidation damage.
Northeastern China is a region of high tick abundance, multiple tick-borne pathogens and likely human infections. The spectrum of diseases caused by tick-borne pathogens has not been objectively evaluated in this region for clinical management and for comparison with other regions globally where tick-transmitted diseases are common. Based on clinical symptoms, PCR, indirect immunofluorescent assay and (or) blood smear, we identified and described tick-borne diseases from patients with recent tick bite seen at Mudanjiang Forestry Central Hospital. From May 2010 to September 2011, 42% (75/180) of patients were diagnosed with a specific tick-borne disease, including Lyme borreliosis, tick-borne encephalitis, human granulocytic anaplasmosis, human babesiosis and spotted fever group rickettsiosis. When we compared clinical and laboratory features to identify factors that might discriminate tick-transmitted infections from those lacking that evidence, we revealed that erythema migrans and neurological manifestations were statistically significantly differently presented between those with and without documented aetiologies (P < 0.001, P = 0.003). Twelve patients (6.7%, 12/180) were co-infected with two tick-borne pathogens. We demonstrated the poor ability of clinicians to identify the specific tick-borne disease. In addition, it is necessary to develop specific laboratory assays for optimal diagnosis of tick-borne diseases.
A comprehensive geochronological and geochemical study was carried out on the gneissic monzogranites, porphyritic granodiorites and charnockites in the Gaozhou complex of the Yunkai massif in the southern part of the South China block to better understand the Early Palaeozoic tectonic regime of the South China block. Laser ablation – inductively coupled plasma – mass spectrometry (LA-ICP-MS) U–Pb dating of zircons indicates an age of 453.2 ± 5.1 Ma to the formation of the gneissic monzogranites, whereas the porphyritic granodiorites and charnockites were generated at 437.0 ± 1.5 Ma and 435.2 ± 2.2 Ma, respectively. The gneissic monzogranites show geochemical features consistent with the high-K, calc-alkaline rock series and are strongly peraluminous. They have SiO2 contents ranging from 67.75 to 69.65 wt. % and relatively low CaO contents (1.66–1.94 wt. %). Their REE patterns are fractionated with enriched LREEs and negative Eu anomalies. The samples also show enrichment in LILEs (e.g. Rb and K) and Pb, and depletion in Sr, Ba and HFSEs (e.g. Nb, Ta, Ti and P). They have εNd(t) values of −8.2 to −7.7. Conversely, the porphyritic granodiorites and charnockites are characterized as medium-K, calc-alkaline rock series and weakly to strongly peraluminous. They exhibit pronounced depletions in HFSEs and positive Pb anomalies. Compared to the earlier gneissic monzogranites, these rocks have relatively lower SiO2 (65.50–69.36 wt. %), but higher CaO contents (3.34–4.05 wt. %), and have slightly lower εNd(t) values (−9.1 to −8.4). Petrography and geochemical compositions of the gneissic monzogranites indicate that they are S-type granite and likely formed by partial melting of Neoproterozoic to Early Palaeozoic immature metagreywackes; whereas The porphyritic granodiorites and charnockites are A-type granite and likely derived from low degrees of partial melting of the dry, granulitic residue depleted by prior extraction of granitic melt. The new data for the Caledonian granitoids in the Yunkai massif suggest that they were formed in a post-collisional tectonic setting. They represent the earliest post-collisional alkaline magmatism reported so far in the Yunkai massif, and thus indicate a tectonic regime switch, from compression to extension, as early as the Late Ordovician to Early Silurian (~450–435 Ma).
This study aimed to investigate the impacts of dietary threonine on intestinal immunity and inflammation in juvenile grass carp. Six iso-nitrogenous semi-purified diets containing graded levels of threonine (3·99–21·66 g threonine/kg) were formulated and fed to fishes for 8 weeks, and then challenged with Aeromonas hydrophila for 14 d. Results showed that, compared with optimum threonine supplementation, threonine deficiency (1) decreased the ability of fish against enteritis, intestinal lysozyme activities (except in the distal intestine), acid phosphatase activities, complement 3 (C3) and C4 contents and IgM contents (except in the proximal intestine (PI)), and it down-regulated the transcript abundances of liver-expressed antimicrobial peptide (LEAP)-2A, LEAP-2B, hepcidin, IgZ, IgM and β-defensin1 (except in the PI) (P<0·05); (2) could up-regulate intestinal pro-inflammatory cytokines TNF-α, IL-1β, IL-6, IL-8 and IL-17D mRNA levels partly related to NF-κB signalling; (3) could down-regulate intestinal anti-inflammatory cytokine transforming growth factor (TGF)-β1, TGF-β2, IL-4/13A (not IL-4/13B) and IL-10 mRNA levels partly by target of rapamycin signalling. Finally, on the basis of the specific growth rate, against the enteritis morbidity and IgM contents, the optimum threonine requirements were estimated to be 14·53 g threonine/kg diet (4·48 g threonine/100 g protein), 15.05 g threonine/kg diet (4·64 g threonine/100 g protein) and 15·17 g threonine/kg diet (4·68 g threonine/100 g protein), respectively.
The prevalence of CHD has been well described worldwide except in Tibet. This study aimed to illustrate the prevalence and composition of CHD in Tibetan children according to altitude.
Methods and results
In the first part, we prospectively recruited 7088 unselected Tibetan children (4–17 years) from south-west Tibet. The total prevalence of CHD increased from 4.6/1000 below 4200 m to 13.4/1000 above 4700 m, with a female-to-male ratio of 1.3:3.1. The total prevalence and female prevalence of patent ductus arteriosus increased more than 10-fold. Females living above 4700 m had exceptionally high prevalence of patent ductus arteriosus (14.9/1000). The prevalence of atrial septal defect was comparable among different altitudes (3.3–3.8/1000). The prevalence of ventricular septal defect was 1.3/1000 below 4700 m, and no cases were found above this altitude. In the second part, we retrospectively reviewed the clinical data of 383 CHD children in Tibet and 73 children at lower altitudes. The percentage of isolated ventricular septal defect decreased from 54.8 to 3.1%, and the percentage of isolated patent ductus arteriosus increased from 8.2 to 68.4% with elevation. Children living below 4200 m (10.4–13.7%) had a larger proportion of complex CHD than those above this altitude (2.0–3.1%). Of the 20 Tibetan children with complex CHD, 14 (70.0%) lived below 4200 m.
A wide variation in CHD prevalence and composition existed in Tibetan children among different altitudes.
Palaeoproterozic metasedimentary rocks, also referred to as khondalites, characterized by Al-rich minerals, are extensively exposed in the nucleus of the Yangtze craton, South China block. Samples of garnet–sillimanite gneiss in the khondalite suite were collected from the Kongling complex for Nd isotopic and elemental geochemical study. These rocks are characterized by variable SiO2 contents ranging from 35.71 to 58.07 wt%, and have low CaO (0.45–0.84 wt%) but high Al2O3 (18.56–29.04 wt%), Cr (174–334 ppm) and Ni (42.5–153 ppm) contents. They have high CIW (Chemical Index of Weathering) values (90.4–94.7), indicating intense chemical weathering of the source material. The samples display light rare earth elements (LREE) enrichment with negative Eu anomalies (Eu/Eu*=0.40–0.68), and have flat heavy rare earth elements (HREE) patterns. The high contents of transition elements (e.g. Cr, Ni, Sc, V) and moderately radiogenic Nd isotopic compositions suggest that the paragneisses might be those of first-cycle erosion products of predominantly mafic rocks mixing with small amounts of felsic moderately evolved Archaean crustal source. Geochemical and Nd isotopic compositions reveal that at least some of the protoliths of Kongling khondalite were sourced from local pre-existing mafic igneous rocks in a continental arc tectonic setting. Combined with documented zircon U–Pb geochronological data, we propose that the Palaeoproterozoic high-pressure granulite-facies metamorphism, rapid weathering, erosion and deposition of the khondalites in the interior of the Yangtze craton might be related to a Palaeoproterozoic collisional orogenic event during 2.1–1.9 Ga, consistent with the worldwide contemporary orogeny, implying that the Yangtze craton may have been an important component of the Palaeoprotorozoic Columbia supercontinent.
The present study investigated the effects of dietary vitamin A on immune function in the proximal intestine (PI), mid intestine (MI) and distal intestine (DI) of young grass carp (Ctenopharyngodon idella). Fish were fed graded levels of dietary vitamin A for 10 weeks, and then a challenge test using an injection of Aeromonas hydrophila was conducted for 14 d. The results showed that, compared with the optimum vitamin A level, vitamin A deficiency significantly decreased fish growth performance, increased enteritis morbidity, decreased intestinal innate humoral immune response and aggravated intestinal inflammation. However, liver-expressed antimicrobial peptide 2A/B mRNA in the DI and IL-6, IL-17D, IL-10, transforming growth factor (TGF)-β1 and TGF-β2 mRNA in the PI were not affected by vitamin A levels. Meanwhile, vitamin A deficiency disturbed inflammatory cytokines in the PI, MI and DI, which might be partly linked to p38 mitogen-activated protein kinase (p38MAPK) signalling and NF-κB canonical signalling pathway (IκB kinase β (IKKβ), IKKγ, inhibitor of κBα, NF-κB p65 and c-Rel) rather than NF-κB non-canonical signalling pathway (NF-κB p52 and IKKα). However, the signalling molecules NF-κB p65 and p38MAPK did not participate in regulating cytokines in the PI. These results suggested that vitamin A deficiency decreased fish growth and impaired intestinal immune function, and that different immune responses in the PI, MI and DI were mediated partly by NF-κB canonical signalling and p38MAPK signalling pathways. On the basis of percentage of weight gain, to protect fish against enteritis morbidity and acid phosphatase activity, the optimum dietary vitamin A levels were estimated to be 0·664, 0·707 and 0·722 mg /kg, respectively.
To describe nutrient intakes, characterize dietary patterns and analyse their associations with sociodemographic characteristics among pregnant women in Shaanxi, China.
Population-based cross-sectional survey.
Twenty counties and ten districts in Shaanxi Province of Northwest China, 2013.
Women (n 7462) were recruited using a stratified multistage random sampling method to report diets during pregnancy, at 0–12 months (median 3 months; 10th–90th percentile, 0–7 months) after delivery.
Pregnant women had higher intakes of fat, niacin and vitamin E than the nutrient reference values, while most micronutrients such as vitamin A, folate, Ca and Zn were reportedly low. Women in the highest education, occupation and household income groups had higher nutrient intakes than those in the lowest groups. Nutrient intake differences also existed by geographic area, residence and maternal age at delivery. Three dietary patterns were identified: balanced pattern, vegetarian pattern and snacks pattern. Participants with high balanced pattern scores tended to be better educated, wealthier, 25–29 years old at delivery, working outside and living in urban areas and central Shaanxi. Women with high scores on the vegetarian pattern and snacks pattern tended to be in low balanced pattern score groups, and had lower nutrient intakes than those in the high balanced pattern score groups.
The study suggested that pregnant women in Shaanxi, China had low intakes of most nutrients such as vitamin A, folate and Ca. Dietary patterns and most nutrient intakes varied by sociodemographic characteristics. Targeted programmes are needed to improve dietary intakes and dietary patterns among sociodemographically disadvantaged groups.
Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a serious hospital and community-acquired infection and some strains are associated with greater severity. We investigated the clinical variability and molecular characteristics of MRSA infections in Shenzhen, China through a study at nine sentinel hospitals from January to December 2014. MRSA infections were classified as community-associated (CA-MRSA), healthcare-associated (HA-MRSA), and healthcare-associated community-onset (HACO-MRSA). In total, 812 MRSA isolates were collected and 183 of these were selected for further study. Patients with HA-MRSA infections were generally of greater age compared to other groups. Distinct body site and clinical presentations were evident in infected patients, e.g. CA-MRSA (skin and soft tissue, 53%), HA-MRSA (respiratory tract, 22%; surgical site, 20%; trauma wounds, 20%) and HACO-MRSA (mastitis, 47%). In contrast to HA-MRSA, other categories of strains were significantly more susceptible to gentamicin, sulfamethoxazole/trimethoprim, and tetracycline. No resistance to vancomycin or linezolid was recorded. The predominant clonal lineage within each strain category was CC59-t437-SCCmec IV/V-agr I (CA, 51·4%; HA, 28·9%; HACO, 52·9%) which exhibited characteristics of a traditional CA clone together with agr I which is more often associated with HA clones. In conclusion, for the three categories of MRSA infections, there were significant differences in clinical characteristics of patients, but the predominant clone in each category shared a similar genetic background which suggests that transmission of MRSA strains has occurred between the community and hospitals in Shenzhen.
The objective of this study was to determine the protective effect of glutamate (Glu) in Cu-induced oxidative injury in fish intestine in vivo and enterocytes in vitro. The results indicated that exposure to 6 mg/l Cu for 72 h induced the production of reactive oxygen species, thereby increasing protein oxidation and lipid peroxidation in enterocytes of grass carp in vitro. Cells exposed to Cu alone resulted in a significant increase in lactate dehydrogenase release, which is accompanied by depletions of antioxidants, including total superoxide dismutase (T-SOD), glutathione S-transferase (GST), glutathione reductase (GR), anti-superoxide anion (ASA), anti-hydroxy radical (AHR) activities and GSH content. Pre-treatment with Glu remarkably prevented the toxic effects of Cu on the T-SOD, GST, GR, AHR, and ASA activities and GSH content in enterocytes. However, Cu induced an adaptive increase in the activities of catalase and glutathione peroxidase (GPx). Glu supplementation further increased GPx activity in enterocytes. Interestingly, the experiment in vivo showed that Glu pre-supplementation significantly elevated SOD, GPx, GST, GR, ASA and AHR activities, as well as GSH content. Further results showed that pre-treatment with Glu could alleviate Cu-induced oxidative injury by elevating antioxidant enzyme activities through regulating the expression of NF-E2-related nuclear factor 2 (Nrf2) mRNA. Together, these results indicated that Glu could attenuate Cu-induced cellular oxidative damage in fish intestine, likely mediated through Nrf2 signalling pathways regulating mRNA expressions of antioxidant enzyme genes and synthesis of GSH.
This study investigated the effects of glycinin on the growth, intestinal oxidative status, tight junction components, cytokines and apoptosis signalling factors of fish. The results showed that an 80 g/kg diet of glycinin exposure for 42 d caused poor growth performance and depressed intestinal growth and function of juvenile Jian carp (Cyprinus carpio var. Jian). Meanwhile, dietary glycinin exposure induced increases in lipid peroxidation and protein oxidation; it caused reductions in superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) activities; and it increased MnSOD, CuZnSOD, GPx1b and GPx4a mRNA levels, suggesting an adaptive mechanism against stress in the intestines of fish. However, dietary glycinin exposure decreased both the activity and mRNA levels of nine isoforms of glutathione-S-transferase (GST) (α, μ, π, ρ, θ, κ, mGST1, mGST2 and mGST3), indicating toxicity to this enzyme activity and corresponding isoform gene expressions. In addition, glycinin exposure caused partial disruption of intestinal cell–cell tight junction components, disturbances of cytokines and induced apoptosis signalling in the distal intestines>mid intestines>proximal intestines of fish. Glycinin exposure also disturbed the mRNA levels of intestinal-related signalling factors Nrf2, Keap1a, Keap1b, eleven isoforms of protein kinase C and target of rapamycin/4E-BP. Interestingly, glutamine was observed to partially block those negative influences. In conclusion, this study indicates that dietary glycinin exposure causes intestinal oxidative damage and disruption of intestinal physical barriers and functions and reduces fish growth, but glutamine can reverse those negative effects in fish. This study provides some information on the mechanism of glycinin-induced negative effects.
The present study was conducted to investigate the anti-inflammatory effect of vitamin D both in juvenile Jian carp (Cyprinus carpio var. Jian) in vivo and in enterocytes in vitro. In primary enterocytes, exposure to 10 mg lipopolysaccharide (LPS)/l increased lactate dehydrogenase activity in the culture medium (P<0·05) and resulted in a significant loss of cell viability (P<0·05). LPS exposure increased (P<0·05) the mRNA expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-8), which was decreased by pre-treatment with 1,25-dihydroxyvitamin D (1,25D3) in a dose-dependent manner (P<0·05). Further results showed that pre-treatment with 1,25D3 down-regulated Toll-like receptor 4 (TLR4), myeloid differentiation primary response gene 88 (Myd88) and NF-κB p65 mRNA expression (P<0·05), suggesting potential mechanisms against LPS-induced inflammatory response. In vivo, intraperitoneal injection of LPS significantly increased TNF-α, IL-1β, IL-6 and IL-8 mRNA expression in the intestine of carp (P<0·05). Pre-treatment of fish with vitamin D3 protected the fish intestine from the LPS-induced increase of TNF-α, IL-1β, IL-6 and IL-8 mainly by downregulating TLR4, Myd88 and NF-κB p65 mRNA expression (P<0·05). These observations suggest that vitamin D could inhibit LPS-induced inflammatory response in juvenile Jian carp in vivo and in enterocytes in vitro. The anti-inflammatory effect of vitamin D is mediated at least in part by TLR4-Myd88 signalling pathways in the intestine and enterocytes of juvenile Jian carp.
The family of interferon-inducible transmembrane proteins (IFITMs) plays a crucial role in inhibiting proliferation, promoting homotypic cell adhesion and mediating germ cell development. In the present study, the full-length cDNAs of zebrafish ifitm1 (744 bp) and ifitm3 (702 bp) were obtained by rapid amplification of cDNA ends (RACE). Reverse transcription polymerase chain reaction (RT-PCR) analysis showed that ifitm1 mRNA was expressed in the ovary, testis, brain, muscle, liver and kidney, while ifitm3 mRNA was only detected in the ovary. Based on in situ hybridization, ifitm1 mRNA was found to be strongly expressed in the ooplasm from stage I to stage II and ifitm3 mRNA was also strongly expressed in the ooplasm from stage I to stage II, furthermore ifitm3 expression ultimately localized to the cortex region beneath the plasma membrane of stage IV oocytes. During development, ifitm1 expression was initially detected in the enveloping layer cells and deep layer cells of shield stage embryos. Then, throughout the segmentation phase (10.25–24 hours post-fertilization (hpf)), ifitm1 expression was mainly detected in the head, trunk and tail regions. Unlike ifitm1, ifitm3 expression was initially detected in sphere stage embryos and was then broadly expressed throughout the embryo from the 70% epiboly stage to 24 hpf. Interestingly, ifitm3 was also expressed in primordial germ cells (PGCs) from the bud stage to 24 hpf. This expression analysis indicates that zebrafish ifitm1 may play a critical role in early organogenesis and may perform immune or hematopoietic functions and ifitm3 might be necessary for PGC migration and the formation of female germ cells.