RNA-Seq reveals miRNA role in thermogenic regulation in brown adipose tissues of goats.

BACKGROUND: MicroRNAs (miRNAs) are a family of short non-coding RNA molecules and play important roles in various biological processes. However, knowledge of the expression profiles and function of miRNAs on the regulation of brown adipose tissue (BAT) thermogenesis remains largely unknown. RESULTS: In this study, we found that brown adipose tissue (BAT) existed within the perirenal fat at 1 day after birth (D1) and transferred into white adipose tissue (WAT) at 30 days after birth (D30) by UCP1 protein expression and immunohistochemistry analysis. After that, we performed RNA sequencing on six libraries of goat BAT and WAT. A total of 238 known miRNAs and 1834 goat novel miRNAs were identified. Moreover, 395 differentially expressed miRNAs including 167 up-regulated and 228 down-regulated miRNAs were obtained in BAT. For the known BAT enriched miRNA, 30 miRNAs were enriched in goat BAT but not in mouse BAT. In addition, miR-433 was enriched in goat BAT but not in mouse BAT. Gain- and loss-of-function experiments reveal that miR-433 reduced the lipid accumulation of brown adipocytes and decreased the expression of BAT marker and mitochondrial related genes. However, miR-433 had no effect on lipid accumulation and thermogenesis in white adipocytes. In addition, miR-433 inhibited the expression of MAPK8 by targeting to the 3'UTR of MAPK8 gene. These data demonstrate that miR-433 acts as a negative regulator in controlling brown adipocytes differentiation and thermogenesis. CONCLUSION: The present study provides a detailed miRNAs expression landscape in BAT and WAT. Furthermore, we found that miR-433, which was highly expressed on BAT had a negative regulatory function on the thermogenesis and adipogenesis in goat brown adipocytes. This study provides evidence for understanding the role of miRNAs in regulating BAT thermogenesis and energy expenditure in goats.

How does national development zone policy affect carbon emissions in China? New evidence from a quasi-natural experiment.

The expansion of China's development zones has made great contributions to economic development, as well as provided practical guidance for other developing countries to implement development zone policies. However, in the context of global advocacy of low carbon, literature about how the development zone policy affect carbon emissions is poor, especially in China at the urban level. Therefore, this study takes China's development zone policy as a quasi-natural experiment, using the panel data of 285 cities in China from 2003 to 2020, and adopting the DID model to analyze its impact on carbon emissions. After a series of robustness tests including placebo test, dynamic test (all independent variables are lagged by one period), endogeneity test, and parallel trend test, the results are basically robust. The findings show that the development zone policy indeed significantly reduces carbon emissions. In addition, we find that cities with higher resource endowments, cities in the eastern and central regions, and other larger cities across the country have better carbon emissions reduction effects. To a certain extent, the research in this paper fills the gap of theoretical research on carbon emissions in terms of the development zone policy, and provides some practical basis for future research in the field of carbon emissions.

[Spatiotemporal variations of ecosystem service values and landscape ecological risk and their correlation in forest area of Loess Plateau, China:A case study of Ziwuling region.] / é»„åœŸé«˜åŽŸæž—åŒºç”Ÿæ€ç³»ç»ŸæœåŠ¡ä»·å€¼ä¸Žæ™¯è§‚ç”Ÿæ€é£Žé™©æ—¶ç©ºå˜åŒ–åŠå ¶å ³è”æ€§­­ä»¥å­åˆå²­åŒºä¸ºä¾‹.

Although the awareness of ecosystem services provided by natural landscape is increa-sing, few studies integrate ecosystem services value (ESV) into ecological risk management and its control. With Ziwuling region as an example, we carried out the gridding resampling of landscape pattern types in the forest area of Ziwuling region in 1980, 1990, 2000, 2010 and 2017. We quantified the ESV and landscape ecological risk and their spatiotemporal variations, based on the 2.5 km×2.5 km grid. The spatial correlation between the ESV and landscape ecological risk was examined. The results showed that the ESV decreased from the center to the outside of Ziwuling region, which increased from 12.345 to 12.633 billion yuan from 1980 to 2017. The landscape ecological risk increased from the center to the outside edge of Ziwuling region, indicating that the landscape ecological risk of Ziwuling region was reduced and the overall ecological situation was improved. There was a significant negative correlation and negative spatial correlation between ESV and landscape ecological risk in this area. The high value-low risk area was mainly located in Ziwu-ling forest area, while the ESV might be maintained in the future.

Oridonin Alleviates LPS-Induced Depression by Inhibiting NLRP3 Inflammasome via Activation of Autophagy.

Objective: Oridonin (Ori) is a diterpene compound that has multiple biological properties. Here, our study was conducted to observe the therapeutic effect of Ori on depression as well as to uncover the mechanism. Methods: Lipopolysaccharide (LPS)-induced depression models were established both in C57BL/6 mice and primary astrocytes, which were treated with Ori, autophagy agonist Rapamycin (Rap) and autophagy inhibitor 3-Methyladenine (3-MA). The depressive-like behaviors were assessed with behavioral tests. Autophagy was evaluated in the hippocampus and astrocytes by investigating autophagosomes under transmission electron microscope (TEM) and detecting LC3II/I, Beclin1 and P62 through western blotting. Astrocyte marker glial fibrillary acidic protein (GFAP) was investigated by immunofluorescence. NLRP3 inflammasome activation was evaluated by detecting IL-1ß, NLRP3, ASC and Caspase-1 expression and reactive oxygen species (ROS) accumulation was quantified via DCFH-DA probe. Autolysosomes, autophagosomes and mitophagy were separately observed through mTag-Wasabi-LC3 plasmid, MitoTracker Deep Red staining, and TEM. Results: Our results showed that Ori administration alleviated LPS-induced depressive-like behaviors and increased GFAP expression in the hippocampus. Furthermore, Ori treatment promoted autophagy activation and cell viability as well as weakened NLRP3 inflammasome activation and ROS accumulation both in LPS-induced mice and astrocytes. Ori promoted the autophagic flux unblocked through enhancing fusion of autophagosomes with lysosomes as well as enhanced mitophagy in LPS-treated astrocytes. The therapeutic effect of Ori was enhanced by Rap and weakened by 3-MA. Conclusion: Collectively, our findings provided a promising antidepressant drug and uncovered that Ori alleviated LPS-induced depression by inhibiting NLRP3 inflammasome through activation of autophagy.

Genome-Wide Identification and Characterization of Long Noncoding RNAs of Brown to White Adipose Tissue Transformation in Goats.

Long noncoding RNAs (lncRNAs) play an important role in the thermogenesis and energy storage of brown adipose tissue (BAT). However, knowledge of the cellular transition from BAT to white adipose tissue (WAT) and the potential role of lncRNAs in goat adipose tissue remains largely unknown. In this study, we analyzed the transformation from BAT to WAT using histological and uncoupling protein 1 (UCP1) gene analyses. Brown adipose tissue mainly existed within the goat perirenal fat at 1 day and there was obviously a transition from BAT to WAT from 1 day to 1 year. The RNA libraries constructed from the perirenal adipose tissues of 1 day, 30 days, and 1 year goats were sequenced. A total number of 21,232 lncRNAs from perirenal fat were identified, including 5393 intronic-lncRNAs and 3546 antisense-lncRNAs. Furthermore, a total of 548 differentially expressed lncRNAs were detected across three stages (fold change ≥ 2.0, false discovery rate (FDR) < 0.05), and six lncRNAs were validated by qPCR. Furthermore, trans analysis found lncRNAs that were transcribed close to 890 protein-coding genes. Additionally, a coexpression network suggested that 4519 lncRNAs and 5212 mRNAs were potentially in trans-regulatory relationships (r > 0.95 or r < -0.95). In addition, Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses showed that the targeted genes were involved in the biosynthesis of unsaturated fatty acids, fatty acid elongation and metabolism, the citrate cycle, oxidative phosphorylation, the mitochondrial respiratory chain complex, and AMP-activated protein kinase (AMPK) signaling pathways. The present study provides a comprehensive catalog of lncRNAs involved in the transformation from BAT to WAT and provides insight into understanding the role of lncRNAs in goat brown adipogenesis.

Glycogen synthase kinase 3ß (GSK3ß) regulates the expression of MyHC2a in goat skeletal muscle satellite cells (SMSCs).

Glycogen synthase kinase beta (GSK3ß) plays an important role in skeletal muscle growth, regeneration, and repair. However, the mechanism of GSK3ß regulating MyHC2a expression is currently not clear. In this study, GSK3ß inhibition promoted skeletal muscle satellite cells (SMSCs) differentiation and increased expression of MyoD, MyoG, MyHC1, and MyHC2a genes. Then we cloned approximately 1.1 kb of goat MyHC2a gene promoter. The deletion fragment (-514/+55) of MyHC2a promoter exhibited the highest level of promoter activity, and a NFATc2 element in this region was responsible for MyHC2a promoter activity. Treatment of SB216713 significantly decreased the transcriptional activity of the fragment (-514/+55). Furthermore, GSK3ß inhibition had no effect on the luciferase activity of MyHC2a promoter after mutating the NFATc2-binding site. These results demonstrated that GSK3ß inhibition promoted SMSCs differentiation and regulated the MyHC2a gene expression through NFATc2 in goat-differentiated SMSCs.

5-Hydr-oxy-1-(3-hydr-oxy-2-naphtho-yl)-3,5-dimethyl-2-pyrazoline.

In the title mol-ecule, C(16)H(16)N(2)O(3), intra-molecular O-H⋯O hydrogen bonds influence the mol-ecular conformation. Inter-molecular O-H⋯O hydrogen bonds [O⋯O = 2.922 (2) Å] link the mol-ecules into centrosymmetric dimers. Weak inter-molecular C-H⋯O inter-actions assemble these dimers into layers parallel to the bc plane.

Expression of NOD1 and downstream factors in placenta, fetal membrane and plasma from pregnancies with premature rupture of membranes and their significance.

OBJECTIVE: To investigate the expression and significance of NOD1 (nucleotide oligomerization domain 1) and its downstream factors in placenta, fetal membrane and plasma of pregnancies with premature rupture of membranes (PROM). METHODS: 60 cases of PROM pregnancies were recruited, including 30 cases of preterm premature rupture of membranes (< 37 weeks) and 30 cases of mature premature rupture of membranes (≥ 37 weeks); 30 healthy pregnancies in the same period were selected as a control group (gestational weeks ≥ 37 weeks). Western blot was used to detect the expression of NOD1, receptor interacting protein 2 (RIP2) and nuclear factor-κB (NF-κB) in placenta and fetal membrane tissues of the three groups. Immunohistochemistry staining was used to investigate the protein levels of NOD1 in the placenta and fetal membrane of three groups of pregnancies. Reverse transcription quantitative PCR (RT-qPCR) demonstrated the expression of NOD1, RIP2, and NF-κB mRNA in placenta, fetal membrane, and plasma of the three groups. The content of NOD1 in plasma was detected by ELISA. RESULTS: Western blot analysis showed that the expressions of NOD1, RIP2 and NF-κB in the placenta and fetal membrane of the preterm PROM group were significantly higher than those in the control group (P < 0.01). Furthermore, the increased levels of NOD1, RIP2 and NF-κB protein in placenta and fetal membrane of mature PROM group were more dramatic than those in the preterm PROM group (P < 0.05). The immunohistochemical staining showed that the staining intensity of NOD1 in placenta and fetal membranes of mature PROM group was stronger those that in the preterm PROM group, and the staining intensity of the former two groups was significantly higher than that of the control group. RT-qPCR detection showed that the mRNA expressions of NOD1, RIP2 and NF-κB in placenta, fetal membrane and plasma of preterm PROM group were significantly higher than in the control group (P < 0.01). In addition, NOD1, RIP2 and NF-κB mRNA levels in the placenta, fetal membranes and plasma of mature PROM group were further increased compared with preterm PROM group (P < 0.01); ELISA assay revealed that the levels of NOD1 in plasma of the mature PROM group, preterm PROM group and control group were (8.34±0.16), (6.82±0.11) and (0.92±0.08) ng/mL, respectively. In the mature PROM group, the content of NOD1 in plasma was increased compared with preterm PROM group (P < 0.05), and plasma NOD1 in both the mature PROM group and preterm PROM group were significantly higher than those in the control group (P < 0.01). CONCLUSIONS: Premature rupture of membranes leads to increaseh mRNA and protein levels of NOD1, RIP2 and NF-κB in placenta, fetal membrane, and peripheral blood, which triggers an inflammatory response and increases the severity of PROM.

N,N'-disalicyloylhydrazine.

The approximately planar mol-ecule of the title compound, C(14)H(12)N(2)O(4), is centrosymmetric and has an E configuration with respect to the N-N bond. This compound adopts the ketoamine form with C=O and C-N distances of 1.233 (3) and 1.331 (4) Å, respectively. Adjacent mol-ecules are assembled into a two-dimensional supra-molecular structure parallel to the (101) plane via inter-molecular O-H⋯O hydrogen bonds.

Neuroligin 1 regulates spines and synaptic plasticity via LIMK1/cofilin-mediated actin reorganization.

Neuroligin (NLG) 1 is important for synapse development and function, but the underlying mechanisms remain unclear. It is known that at least some aspects of NLG1 function are independent of the presynaptic neurexin, suggesting that the C-terminal domain (CTD) of NLG1 may be sufficient for synaptic regulation. In addition, NLG1 is subjected to activity-dependent proteolytic cleavage, generating a cytosolic CTD fragment, but the significance of this process remains unknown. In this study, we show that the CTD of NLG1 is sufficient to (a) enhance spine and synapse number, (b) modulate synaptic plasticity, and (c) exert these effects via its interaction with spine-associated Rap guanosine triphosphatase-activating protein and subsequent activation of LIM-domain protein kinase 1/cofilin-mediated actin reorganization. Our results provide a novel postsynaptic mechanism by which NLG1 regulates synapse development and function.

p21-activated kinase 1 restricts tonic endocannabinoid signaling in the hippocampus.

PAK1 inhibitors are known to markedly improve social and cognitive function in several animal models of brain disorders, including autism, but the underlying mechanisms remain elusive. We show here that disruption of PAK1 in mice suppresses inhibitory neurotransmission through an increase in tonic, but not phasic, secretion of endocannabinoids (eCB). Consistently, we found elevated levels of anandamide (AEA), but not 2-arachidonoylglycerol (2-AG) following PAK1 disruption. This increased tonic AEA signaling is mediated by reduced cyclooxygenase-2 (COX-2), and COX-2 inhibitors recapitulate the effect of PAK1 deletion on GABAergic transmission in a CB1 receptor-dependent manner. These results establish a novel signaling process whereby PAK1 upregulates COX-2, reduces AEA and restricts tonic eCB-mediated processes. Because PAK1 and eCB are both critically involved in many other organ systems in addition to the brain, our findings may provide a unified mechanism by which PAK1 regulates these systems and their dysfunctions including cancers, inflammations and allergies.

Association of Mannose-binding Lectin Polymorphisms with Tuberculosis Susceptibility among Chinese.

Tuberculosis (TB) is caused by infection of Mycobacterium tuberculosis. Host genetic variability is an important determinant of the risk of developing TB in humans. Although the association between MBL2 polymorphisms and TB has been studied in various populations, the results are controversial. In this study four functional single-nucleotide polymorphisms (SNPs, H/L, X/Y, P/Q and A/B) across the MBL2 gene were genotyped by direct DNA sequencing of PCR products in a case-control population of Chinese Han origin, consisting of 1,020 patients with pulmonary TB and 1,020 controls. We found that individuals carrying variant allele at A/B (namely BB or AB genotypes) was associated with increased susceptibility to TB (odds ratios [OR] = 1.57, 95% confidence interval [CI] 1.30-1.91, P = 1.3 × 10-6). Additionally, LYPB haplotype showed a significant association with increased risk of TB (OR = 1.54, 95% CI 1.27-1.87, P = 4.2 × 10-6; global haplotype association P = 3.5 × 10-5). Furthermore, individuals bearing low- or medium- MBL expression haplotype pairs had an increased risk of TB (OR = 1.56, 95% CI 1.29-1.90, P = 1.4 × 10-6). Thus, the reduced expression of functional MBL secondary to having MBL2 variants may partially mediate the increased susceptibility to TB risk.

Exceedingly expanded retroauricular flaps for microtia reconstruction.

BACKGROUND: The authors propose here a new strategy to obtain exceedingly expanded retroauricular mastoid skin for sufficient coverage of the three-dimensional autogenous costal cartilage framework generally used in auricular reconstruction surgery. From February 2000 to September 2009, 42 microtia reconstructions were performed using this new strategy. METHODS: Auricular reconstruction was performed in three surgical stages. In the first stage, a 50-ml kidney-shaped expander was inserted subcutaneously in the retroauricular mastoid region. From 5 to 8 ml saline was then injected into the expander every 4 days until the final volume of the expander reached 100-120 ml. In the second stage, we divided the expanded mastoid skin into a superior two-third region (flap A) and an inferior one-third region (flap B, rotation flap). Autogenous costal cartilage framework was then enveloped by these expanded flaps. Tragus construction and conchal excavation was performed in the third stage. RESULTS: All patients were followed up from 6 months to 4 years after reconstruction. A total of 36 cases reported to be satisfied with the appearance of good shape, accurate size, right orientation, and duplication of well-detailed structures. Further revision was requested by six of the total. Complications in this series includes one case of haematoma, two cases of partial evection of the expanded skin and two cases of partial skin necrosis of the helix. All the complications were treated appropriately. CONCLUSION: Exceeding expansion can provide sufficient retroauricular non-hair-bearing skin tissues for draping the auricular cartilage framework. Skin grafts and retroauricular fascial flap are not needed any more. Patients are usually satisfied with their reconstructive auricles as regards the size, location, projection, convolution, skin-colour matching, etc. Exceedingly expanded retroauricular flaps are the appropriate envelope for the auricular cartilage framework.