Neuron-secreted NLGN3 ameliorates ischemic brain injury via activating Gαi1/3-Akt signaling.

We here tested the potential activity and the underlying mechanisms of neuroligin-3 (NLGN3) against ischemia-reperfusion-induced neuronal cell injury. In SH-SY5Y neuronal cells and primary murine cortical neurons, NLGN3 activated Akt-mTOR and Erk signalings, and inhibited oxygen and glucose deprivation (OGD)/re-oxygenation (OGD/R)-induced cytotoxicity. Akt activation was required for NLGN3-induced neuroprotection. Gαi1/3 mediated NLGN3-induced downstream signaling activation. NLGN3-induced Akt-S6K1 activation was largely inhibited by Gαi1/3 silencing or knockout. Significantly, NLGN3-induced neuroprotection against OGD/R was almost abolished by Gαi1/3 silencing or knockout. In vivo, the middle cerebral artery occlusion (MCAO) procedure induced NLGN3 cleavage and secretion, and increased its expression and Akt activation in mouse brain tissues. ADAM10 (A Disintegrin and Metalloproteinase 10) inhibition blocked MCAO-induced NLGN3 cleavage and secretion, exacerbating ischemic brain injury in mice. Neuronal silencing of NLGN3 or Gαi1/3 in mice also inhibited Akt activation and intensified MCAO-induced ischemic brain injury. Conversely, neuronal overexpression of NLGN3 increased Akt activation and alleviated MCAO-induced ischemic brain injury. Together, NLGN3 activates Gαi1/3-Akt signaling to protect neuronal cells from ischemia-reperfusion injury.

Integrated analysis of microRNA and mRNA interactions in ovary of counter-season breeding and egg-ceased geese (Anser cygnoides).

Egg-ceasing is a phenomenon that occurs in most avian species and significantly reduces productivity. Although several factors are reported to regulate the reproduction progress, the underlying molecular mechanism of egg-ceasing remains obscure. Herein, we identified and explored the differentially expressed miRNAs and mRNAs involved in ovarian atrophy via high throughput sequencing. We identified a total of 901 mRNAs and 50 miRNAs that were differentially expressed in egg-laying and atrophic ovaries. Among them, numerous differentially expressed gene (DEG) transcripts and target genes for miRNAs were significantly enriched in Gene Ontology terms such as reproductive processes, cell proliferation, and apoptosis pathways. In addition, an interaction network was constructed by considering target relationships and correlation of the expression levels between ovary development-related genes, miRNAs and pathways. We discovered mRNA and miRNAs transcripts that are candidate regulators of ovary development in egg-ceased geese. Our findings expanded our understanding of the functional of miRNAs in ovarian atrophy and demonstrated that RNA-Seq is a powerful tool for examining the molecular mechanism in regulating egg-ceasing.

Genotype frequency distributions of 28 SNP markers in two commercial lines and five Chinese native chicken populations.

BACKGROUND: Modern breeding in the poultry industry mainly aims to produce high-performance poultry lines and breeds in two main directions of productivity, meat and eggs. To understand more about the productive potential of lowly selected Chinese native chicken populations, we selected 14 representative SNP markers strongly associated with growth traits or carcass traits and 14 SNP markers strongly associated with egg laying traits through previous reports. By using the MassArray technology, we detected the genotype frequency distributions of these 28 SNP markers in seven populations including four lowly selected as well as one moderately selected Sichuan native chicken populations, one commercial broiler line and one commercial layer line. RESULTS: Based on the genotype frequency distributions of these 28 SNP markers in 5 native chicken populations and 2 commercial lines, the results suggested that these Chinese indigenous chicken populations have a relatively close relationship with the commercial broiler line but a marked distinction from the commercial layer line. Two native chicken breeds, Shimian Caoke Chicken and Daheng Broilers, share similar genetic structure with the broiler line. CONCLUSIONS: Our observations may help us to better select and breed superior domestic chickens and provide new clues for further study of breeding programs in local chicken populations.

[Genetic regulation of broodiness in poultry].

Broodiness is a behavior commonly occurring in the poultry industry, which is characterized by inappetence, egg-laying cessation and incubation. Different from laying fowls, the ovary and oviduct of broodiness fowls is degenerate. Broodiness is a low heritability trait, which is controlled by multiple genes in autosomes and influenced by three factors, including environment, endocrine and genetics. In addition to the observation of behavioral characteristics, the current research of broodiness focuses on evaluating the genetic mode, endocrine factors, nesting candidate genes and their polymorphisms in poultry. Given the rapid development of high-throughput sequencing, the joint analyses of genomics, transcriptomics and proteomics have been used to screen out the candidate genes, pathways and molecular mechanism of broodiness. In this review, we summarize the candidate genes, microRNAs and pathways involved in broodiness to provide a reference for further research on poultry broodiness.

KLF5 functions in proliferation, differentiation, and apoptosis of chicken satellite cells.

KLF5 is an important regulator of cell proliferation, differentiation, and apoptosis in mammals. Little is known about the function of KLF5 in the regulation of chicken. Hence, qPCR was used to detect the expression of KLF5 in different tissues of chicken. And chicken skeletal muscle satellite cells (SMSCs) were transfected KLF5-specific small interfering RNA (siRNA) to assay SMSCs' proliferation, differentiation, and apoptosis. The results showed that KLF5 expressed higher in skeletal muscle than in the other tissues of chicken. Knockdown of KLF5 significantly inhibited the differentiation and increased apoptosis of chicken SMSCs, but it had no significant effect on proliferation of SMSCs. These results indicate that KLF5 plays an essential role during myogenesis, which will affect muscle repair and muscle regeneration, and may ameliorate muscle aging or sarcopenia.

Molecular Characterization, Expression and Functional Analysis of Chicken STING.

Innate immunity is an essential line of defense against pathogen invasion which is gained at birth, and the mechanism involved is mainly to identify pathogen-associated molecular patterns through pattern recognition receptors. STING (stimulator of interferon genes) is a signal junction molecule that hosts the perception of viral nucleic acids and produces type I interferon response, which plays a crucial role in innate immunity. However, relatively few studies have investigated the molecular characterization, tissue distribution, and potential function of STING in chickens. In this study, we cloned the full-length cDNA of chicken STING that is composed of 1341 bp. Sequence analyses revealed that STING contains a 1140-bp open-reading frame that probably encodes a 379-amino acid protein. Multiple sequence alignments showed that the similarity of the chicken STING gene to other birds is higher than that of mammals. Real-time polymerase chain reaction (PCR) assays revealed that STING is highly expressed in the spleen, thymus and bursa of fabricious in chickens. Furthermore, we observed that STING expression was significantly upregulated both in vitro and in vivo following infection with Newcastle disease virus (NDV). STING expression was also significantly upregulated in chicken embryo fibroblasts upon stimulation with poly(I:C) or poly(dA:dT). Taken together, these findings suggest that STING plays an important role in antiviral signaling pathways in chickens.

Compound Tufuling Granules ([characters: see text]) regulate glucose transporter 9 expression in kidney to influence serum uric acid level in hyperuricemia mice.

OBJECTIVE: To explore the effect of Compound Tufuling Granules ([characters: see text], CTG) on regulating glucose transporter 9 (GLUT9) expression in the kidney to influence the uric acid excretion by the kidney and serum uric acid (SUA) level in hyperuricemia mice. METHODS: Sixty Kunming male mice were randomly divided into the control group, model group, benzbromarone group, and CTG high-, middle- and low-dose groups. The yeast extract and uricase inhibition method were used to build hyperuricemia model, and the corresponding drugs were administrated on the 7th day. On the 21st day the 24-h urine was collected, on the 22nd day the blood was collected, the SUA level was detected by uricase colorimetry, and the mRNA and protein expressions of GLUT9 were detected by quantitative real-time polymerase chain reaction and Western blot, respectively. RESULTS: Compared with the model group, the levels of SUA and the mRNA and protein expressions of GLUT9 were significantly decreased, and the fraction excretion of uric acid (FEUA) was significantly increased in the CTG groups and benzbromarone group (all P<0.05). There was no significant difference in the above indicators between the CTG high-dose group and benzbromarone group (P>0.05). SUA is positively related to the GLUT9 mRNA and protein expressions in the kidney (P<0.05 or P<0.01). CONCLUSIONS: CTG can significantly reduce the SUA and increase the FEUA. In addition, CTG can effectively inhibit the mRNA and protein expressions of GLUT9 in the kidney of hyperuricemia mice to inhibit the uric acid re-absorption, promote uric acid excretion and reduce SUA.

Effects of Xie-Zhuo-Chu-Bi-Fang on miR-34a and URAT1 and their relationship in hyperuricemic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Xie-Zhuo-Chu-Bi-Fang (XZCBF) is an empirical formula that was developed based on the principles of traditional Chinese medicine, for the therapeutic purpose of treating hyperuricemia. XZCBF has been clinically utilized in the Department of Traditional Chinese Medicine at General Hospital of Guangzhou Military Command of PLA for many years and has exhibited favorable efficacy. The aim of the study is to evaluate the effects of XZCBF on the expression of uric acid transporter 1 (URAT1) and miR-34a in hyperuricemic mice and to determine, the correlation between the two expression levels. MATERIALS AND METHODS: A hyperuricemic animal model was created by administering adenine and allantoxanic acid potassium salt to mice. The blood uric acid levels were measured in these model mice after treatment with XZCBF for 15 days. The potential targets of miR-34a were screened. The expression levels of miR-34a and URAT1 in the renal tissues collected from the model mice were determined by quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and their correlation was further established by immunohistochemistry and in situ hybridization. RESULTS: The uric acid levels in the model mice were significantly higher than those in the blank controls (P<0.05). These levels were significantly lower in the three groups receiving different doses of XZCBF (P<0.05), which was, in agreement with the downregulation of URAT1 and the upregulation of miR-34a in each group. The mRNA expression level of URAT1 was positively correlated with the concentration of uric acid but, negatively correlated with the expression level of miR-34a. CONCLUSIONS: The ability of XZCBF to facilitate the excretion of uric acid and to lower its level in the model group was mediated by the upregulation of miR-34a and the inhibition of URAT1 mRNA expression, which suggests that XZCBF could be an option for the treatment of hyperuricemia in mice.

[Assessment on the clinical efficacy and safety of xiezhuo chubi recipe in treating hyperuricemia].

OBJECTIVE: To observe the clinical efficacy and safety of Xiezhuo Chubi Recipe (XCR) on hyperuricemic patients. METHODS: 99 patients with hyperuricemia were randomly assigned to the XCR group, the Benzbromarone group, and the blank control group. Patients in the XCR group took XCR, one dosage daily, twice per day. Patients in the Benzbromarone group took Benzbromarone Tablet (50 mg each tablet, once per day). Patients in the blank control group were not treated with any drug, but only with clinical observation. Twenty days consisted of one course of treatment. The laboratory data including uric acid, blood routines, urine routines, the liver function, and the renal function were statistically analyzed before and after treatment. RESULTS: The blood uric acid decreased in the three groups after treatment (P<0.05). The total effective rate was 85.71% in the XCR group, 92.86% in the Benzbromarone group, and 23.33% in the blank control group. There was no statistical difference between the XCR group and the Benzbromarone group (P>0.0167). There was no significant difference in the safety indices such as blood routines, urine routines, liver functions, and renal functions of the XCR group between before and after treatment (P>0.05). CONCLUSION: XCR could effectively reduce the uric acid level with higher safety.

[Effects of cleaners on the color stability of prosthesis silicone rubbers].

OBJECTIVE: To evaluate the effect of different cleaners on the color stability of two silicone rubbers used for maxillofacial prosthesis, and to provide recommendations for clinical use. METHODS: Thirty skin-color columniform specimens (12 mm diameter, 10 mm height) of two silicone rubber (A:A-2000; Z:ZY-1) were prepared, randomly divided into 6 groups according to the table of random number, and cleaned with the following solutions: isopropyl alcohol (I), three kinds of denture cleaners (P: Polident, S: Steradent, C: Cleansoft) and distilled water (D), simulating the total immersion time of 1 year (1, 15, 10, 3 and 10 min each time respectively). Control group was kept in dark place without treatment. The L(*), a(*), b(*) value were tested before and after immersion. Then color difference value was calculated. RESULTS: Color differences were different among groups. Color difference in group I (A: 2.15, Z: 2.00) were significantly greater than that in any other group. There were no significant differences between groups using denture cleaner P (A: 0.36, Z: 0.36), C (A: 0.42, Z: 0.37) and S (A: 0.33, Z: 0.38), and group D (A: 0.22, Z: 0.23). CONCLUSIONS: Isopropyl alcohol causes the most severe fading, and denture cleaners and distilled water cause obscure fading.

MicroRNA expression patterns of the kidney in hyperuricemia mice treated with Xiezhuo Chubi Decoction.

OBJECTIVE: To investigate the effects of Xiezhuo Chubi Decoction (XZCBD) on the microRNA expression patterns of kidney in mice with hyperuricemia. METHODS: Sixty Kunming male mice were randomly divided into the high-, medium-, and low-dose XZCBD groups, benzbromarone group, model group, and control group. Except the control group, all mice were established with yeast method combined with uricase inhibition method to build hyperuricemia model, and the corresponding drugs (37.5 g/kg, 18.75 g/kg, 9.375 g/kg, and 0.02 g/kg per day) were administrated on the 7th day. On the 22nd day, the blood uric acid concentration was detected, and microRNA with obvious changes in kidney was screened with qRT-PCR. RESULTS: The uric acid in the model group was higher than that in the control group, and the levels of the uric acid were reduced after being treated with XZCBD; the differences among groups were significant (P<0.05). Compared with the control group, 32 kinds of microRNA expression changes were detected on the 15th day after being treated with high-dose XZCBD by microRNA expression profile screening. Among them, miR-34a could inhibit the expression of human urate anion exthanger 1, and miR-146a might have inhibited the inflammatory reaction. CONCLUSION: XZCBD could significantly reduce the serum uric acid level; its effect on hyperuricemia might be through affecting microRNA expressions.