Chemokine receptor 7 mediates miRNA-182 to regulate cerebral ischemia/reperfusion injury in rats.

AIMS: Chemokine receptor 7 (CXCR7) exerts protective effects on the brain. MicroRNAs (miRNAs) are involved in cerebral ischemia/reperfusion (I/R) injury, but their involvement in CXCR7-mediated brain protection is unknown. In this study, we investigated the role of miRNAs in CXCR7-mediated brain protection. METHODS: CXCR7 levels in peripheral blood samples from patients with acute ischemic stroke (AIS) and ischemic penumbra area brain tissues from middle cerebral artery occlusion (MCAO) rats after recanalization were measured. An miRNA microarray analysis was performed to examine the expression of miRNAs caused by CXCR7 knockdown in ischemic penumbra area brain tissue in middle cerebral artery occlusion-reperfusion rats and to predict corresponding downstream target genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed the most enriched pathways. A dual-luciferase reporter assay confirmed the direct regulation of miR-182 on the target gene TCF7L2. The correlation between TCF7L2 and CXCR7/miR-182 was verified using rescue assays. RESULTS: CXCR7 expression was upregulated in MCAO rats and mechanical thrombectomy patients with AIS compared to that in controls. The motor and sensory functions of MCAO rats with CXCR7 knockdown further decreased, and the infarct volume and cerebral edema increased. miRNA microarray data showed that seven miRNAs were differentially expressed after shRNA-CXCR7 treatment. The dual-luciferase reporter assay confirmed that miR-182 directly targeted the TCF7L2 gene. Rescue assays confirmed that TCF7L2 is downstream of CXCR7/miR-182. KEGG pathway analysis showed that the Hippo pathway may be a key pathway in CXCR7 upregulation and plays a role in protecting the brain after interventional surgery. Animal experiments have shown that CXCR7-mediated cerebral I/R injury promotes the phosphorylation of key molecules YAP and TAZ in the Hippo pathway. CONCLUSION: CXCR7 protects against cerebral I/R injury, possibly via the miR-182/TCF7L2/Hippo pathway. These results indicate that CXCR7 affects cerebral ischemia-reperfusion injury through miRNA regulation and downstream pathways.

How Information Repertoire Affects Vaccine Hesitancy: Processes of Information Verification and Cognitive Elaboration.

As information consumption plays a critical role in addressing vaccine hesitancy in the hybrid media environment, it becomes crucial to understand how individuals' use of a combination of channels and sources affects their vaccine hesitancy. Based on information repertoire approaches emphasizing the multiplicity of channels and sources, we investigated different patterns of information repertoire related to the COVID-19 pandemic and how these patterns affected vaccine hesitancy through different informational mechanisms. Our results based on a U.S. sample suggest that while a richer information repertoire related to increased confidence in vaccines through increased information verification, this richness also corresponded with deepened vaccine hesitancy through heightened cognitive elaboration and perceived information inconsistency. Our findings support the utility of repertoire approaches for better understanding health information acquisition in the complex media ecology.

Adsorption of water on the pristine and defective semiconducting 2D CrPX3monolayers (X: S, Se).

The effect of vacancy and water adsorption on the electronic structure of semiconducting 2D trichalcogenide material CrPX3(X: S, Se) is studied using state-of-the-art density functional theory (DFT) approach. It is found that chalcogen vacancies play a minor role on the electronic structure of CrPX3in the vicinity of the Fermi level leading to the slightly reduced band gap for these materials, however, inducing strongly localised defect states which are placed in the energy gap formed by the valence band states. Our DFT calculations show that the interaction of water molecules with CrPX3, pristine and defective, can be described as physisorption and the adsorption energy for H2O is insensitive to the difference between pristine and chalcogen-defective surface of trichalcogenide material. These results are the first steps for the theoretical description of the ambient molecules interaction with 2D semiconducting CrPX3material, that is important for its future experimental studies and possible applications.

Gas-solid interfacial charge transfer in volatile organic compound detection by CuCrO2nanoparticles.

Nanostructured metal oxide semiconductors have received great attention used as the chemiresistive layer of gas sensor to detect the volatile organic compound recently. As indispensable complementary parts for dominative n-type semiconductors, the p-type metal oxides based gas sensors fail to be studied sufficiently, which hampers their practical applications. In this work, the p-type delafossite CuCrO2nanoparticles were synthesized, characterized, and tested for gas sensing, followed by the first principles calculations to simulate the generation of chemiresistive signal. The hydrothermal synthesis time of CuCrO2nanoparticles is optimized as 24 h with a higher proportion of oxygen vacancies but a smaller size, which is confirmed by the microscopy and spectrum characterization and allows for a prevailing gas sensitivity. Meanwhile, this CuCrO2gas sensor is proven to perform a higher selectivity to n-propanol and a low detection limit of 1 ppm. The adsorption sites and charge variations of dehydrogenation at the gas-solid interface predicted by the theoretical analysis are claimed to be crucial to such selectivity. It is an innovative approach to understand the chemiresistive gas sensing by evaluating the preference of charge transfer between the sensor and target gaseous molecule, which provides a new route to precisely design and develop the advanced sensing devices for the diverse applications.

Potential role of miR-155-5p in fat deposition and skeletal muscle development of chicken.

miR-155 has multiple functions in many physiological and pathological processes. However, little is known about the expression characteristics of avian miR-155. In the present study, partial pri-miR-155 sequences were cloned from AA+ broiler, Sanhuang broiler and Hy-Line Brown layer, respectively. Stem-loop qRT-PCR was performed to detect the miR-155-5p spatiotemporal expression profiles of each chicken breed, and the target genes of miR-155-5p were predicted in Gene Oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The results showed that the partial pri-miR-155 sequences of different breeds of chicken were high conserved. The expression patterns of miR-155-5p between broiler and layer were basically similar, and miR-155-5p was expressed highly in immune related tissues (spleen, thymus and bursa). In the same old chicken (14 days old), miR-155-5p expression activity of fat tissue all had higher level in the three chicken breeds, but the expression activities in skeletal muscle of broilers were significantly lower than that of layer (P<0.05). In different development stages of Hy-Line Brown layer, miR-155-5p expression activities in skeletal muscle of 14-day-old and 10-month-old layers were significantly lower than that of 24-month-old layer (P<0.05). Fat related target genes (ACOX1, ACOT7, FADS1, SCD and HSD17B12) and skeletal muscle related target genes (CCNT2, DMD, CFL2, MAPK14, FLNB, ZBTB18 and CDK5) of miR-155-5p were predicted, respectively. The results indicate that miR-155-5p may be an important factor inhibiting the fat deposition and skeletal muscle development in chicken.

[Expression and function characterization of Gimap5 gene from different breeds of broilers].

GTPase immune-associated protein 5 (Gimap5), a key factor in maintaining T cell homeostasis, plays important roles in immune and inflammatory processes. However, its function and characteristics in poultry have not been reported. In this study, AA⁺ and Sanhuang broilers were used as models. The full-length coding sequence of the Gimap5 gene was cloned by RT-PCR and analyzed using bioinformatic methods. Tissue expression and distribution characteristics of the Gimap5 gene and its functional characteristics in inflammatory response were analyzed by RT-qPCR, respectively. The full-length coding sequences of the Gimap5 gene from AA⁺ and Sanhuang broilers were 771 bp, encoding 256 amino acids, and presented low conservative among different species. The AIG-1 domain of N-terminal and alpha-helix structure of C-terminal transmembrane sequence of might play important roles in the Gimap5 protein function and cell localization, respectively. The Gimap5 gene was widely distributed and expressed in various tissues and the pattern of its expression change was basically similar between AA⁺ and Sanhuang broilers. However, there were some differences in expression activity between the two breeds or various tissues of the same breed. In inflammatory response, the expression activities of the Gimap5 gene were down-regulated in blood and liver (P<0.05), but up-regulated in the bursa of Fabricius (P<0.01). It is speculated that Gimap5 is a multifunctional gene involved in the development of the body and inflammatory response, and has potential application value for diagnostic marker of inflammatory response.

Microwave-assisted Synthesis of N,S-co-carbon Dots as Switch-on Fluorescent Sensor for Rapid and Sensitive Detection of Ascorbic Acid in Processed Fruit Juice.

To achieve a rapid, sensitive, and economical method for the detection of ascorbic acid (AA) in the presence of Fe3+, a nitrogen and sulfur co-doped carbon dots (N,S-co-CDs) based fluorescence sensing system was developed. In this work, N,S-co-CDs were successfully synthesized via a one-step microwave-assisted method within 2.5 min using ammonium citrate and L-cysteine as precursors. The fluorescence of N,S-co-CDs was quenched (off ) by Fe3+ through a static-quenching mechanism. Subsequently, the fluorescence was recovered (on) after introducing AA into the quenched system, which was attributed to the reduction effect of AA for Fe3+. Therefore, a switch-on sensor (N,S-co-CDs/Fe3+ system) was developed for AA detection. Under optimal conditions, the limit of detection (LOD) of 2.31 µmol/L for AA was obtained over a linear range from 0 to 150 µmol/L. Furthermore, the proposed sensing method was successfully applied to detect AA in processed fruit juice with satisfactory results. The most important is that the sensor derived from a microwave-assisted method has simple and eco-friendly synthesis processes, is rapid, and has high detection efficiency. Therefore, such a switch-on sensor may be a promising candidate sensor for AA detection in processed fruit samples.

Exploring the expression and preliminary function of chicken Gimap5 gene.

GTPase immune-associated protein 5 (Gimap5) plays a key role in maintaining T cell homeostasis, immunological tolerance and inflammatory processes. However, there are no reports on the chicken Gimap5 gene. In this study, the Gimap5 gene was first cloned from chicken and characterized its tissue expression characteristics in different developmental stages. The transcriptional activities of the Gimap5 gene in immune response were identified. The results showed that full-length cDNA sequence of Gimap5 contained 771 bp and encoded a 256-amino acid protein. The Gimap5 gene was transcribed in various tissues and different development stages. The transcriptional activities of Gimap5 gene in the most tissues increased with the development of chicken, but significantly up to peak in liver and large intestine of 10-month-old chicken. The Gimap5 gene exhibited differential transcriptional activities in immune-related tissues in immune responses, with down-regulated in liver (P < 0.01), spleen (P < 0.05) and bursa of Fabricius (P < 0.05), and up-regulated in thymus (P < 0.01). The results show that Gimap5 may be a multifunctional gene involved in tissue function, development and immune response in chicken. These data can provide the foundation for further study of Gimap5.

Arih2 gene influences immune response and tissue development in chicken.

Ariadne homolog 2 (ARIH2), as an E3 ubiquitin ligase, is one of the important factors involved in regulating biological functions, such as inflammation and skeletal muscle degeneration. In the present study, the full-length coding sequence of Arih2 gene was cloned from Hy-Line Brown chicken. The tissue transcriptional profiles of Arih2 gene at different developmental stages were detected using quantitative real-time PCR (qRT-PCR), and the Arih2 functional characteristics in immune response were analyzed. The results showed that the full-length coding sequence of Arih2 gene was 1473 bp, encoding 490 amino acids, and conservative between different species. The Arih2 gene was transcribed in various tissues at different developmental stages, and its transcriptional activities varied significantly between multiple tissues. With the development of chicken, Arih2 gene was basically up-regulated in heart, liver, kidney, skeletal muscle and glandular stomach, but fluctuated significantly in large intestine. In immune response, the transcriptional activities of Arih2 gene exhibited significant changes in the bursa, thymus and blood (P<0.05). The results showed that Arih2 might be a multifunctional gene involved in tissue development and immune response in chicken, and have a potential possible application as diagnostic marker for identifying immune response.

Nutrients, phytochemicals and antioxidant activities of 26 kidney bean cultivars.

Detailed characterization in nutrients and phytochemicals with antioxidant activities of 26 kidney beans was performed. The kidney beans contained high levels of dietary fiber (29.32-46.77%), resistant starch (9.16-18.09%) and protein (22.06-32.63%) but low levels of lipid (1.05-2.83%) and sugars (1.55-9.07%). The monosaccharide composition of soluble fiber was dominated by arabinose, galactose, mannose and galacturonic acid. The ratio of essential amino acid to the total amino acid was ranged from 0.29 to 0.36. The predominant fatty acid was polyunsaturated fatty acids, accounting for 47.54-67.26% of total fatty acids. The total tocopherol content was in the range of 12.83-68.35 µg/g, predominantly γ-tocopherol, followed by δ-tocopherol. In addition, certain levels of total phenolics and flavonoids with respective values of 0.25-3.79 mg gallic acid equivalent/g dry weight and 0.19-7.05 mg rutin equivalent/g dry weight resulted in significant antioxidant activities. And a good correlation was observed between TPC and FRAP values (R2 = 0.8030). The results indicated that kidney beans are excellent sources of health-promoting compounds.

[Water extract from Codonopsis thalictrifolia wall affects the reproductive system of male infant rats].

OBJECTIVE: To study the impact of the water extract from Codonopsis thalictrifolia Wall (CTW) on the reproductive METHODS: We divided 32 male SD infant rats into four groups of equal number to be treated intragastrical-system of male infant rats. ly with distilled water (control) and CTW at 10 g/kg (low dose) , 20 g/kg (medium dose), and 40 g/kg (high dose), respectively, twice a day for 2 weeks. Then we killed the rats, measured the levels of testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum, obtained the testis weight, body weight, testis visceral coefficient and sperm concentration, and detected sperm viability, sperm motility and the level of cyclic adenosine monophosphate (cAMP) in the Leydig cells, followed by RESULTS: Compared with the control group, the low-dose, me-analysis of differences among different groups using the SPSS software. Medium-dose and high-dose CTW groups showed significant decreases in the serum T level ([3.09 +/-0.42] vs [1.22 +/-0. 32] , [1.06 +/- 0.29] and [0.57 +/-0.18] nmol/L, P<0.01), testis weight ([1.40 +/-0.16] vs [0.96 +/-0.09], [0.92 +/-0.11] and [0.91 +/- 0.08] g, P <0.01), and sperm concentration ([1.03 +/-0.16] vs [0.19 +/-0.07], [0.17 +/-0.08] and [0.16 +/-0.07] x 10(6)/ml, P <0.01), but a dramatic elevation in the testis visceral coefficient ([42.22 +/- 3.02] vs [51.39 +/- 3.09], [52.28 +/- 4.86] and [54.13 +/-6.06] mg/10 g, P <0.01); the medium- and high-dose CTW groups exhibited remarkable increases in the levels of serum LH ([13.62+/-0.89] vs [14.69 +/-0.12] and [14.93 +/-0.28] ng/L, P<0.01) and FSH ([4.32 +/-0.18] vs [4.77 +/-0.23] and [4.89 +/-0. 38] IU/L, P <0.05); all the three CTW groups showed markedly inhibited serum T secretion ([1.85 +/- 0.18] vs [1.42 +/-0.15], [1.12+/-0.18] and [0.88 +/-0.21] nmol/L, P<0.01) and intracellular cAMP ([5.51 +/-0.12] vs [4.39+/-0.06], [4.28 +/-0.07] and [4.11 +/- 0.10] nmol/L, P <0.01) in the Leydig cells. CONCLUSION: The water extract from CTW may reduce the synthesis of testosterone in the serum of male infant rats through the PKA pathway and consequently inhibit their testicular development and sperm production and affect the development of their reproductive system.

Metabolic profiling of Gynostemma pentaphyllum extract in rat serum, urine and faeces after oral administration.

Folk drug Gynostemma pentaphyllum (Thunb.) Makino contains many biologically active phytochemicals which have been demonstrated to be effective against chronic diseases. As in vivo anti-tumor experiments of G. pentaphyllum extract (GP) show much stronger antitumor activities than in vitro, it is important and necessary to understand the metabolic study of GP. A sensitive and specific U-HPLC-MS method was utilized for the first time to rapidly identify gypenosides and its possible metabolites in rat serum, urine, and faeces after oral administration. Solid phase extraction was utilized in the sample preparation. Negative Electrospray ionisation (ESI) mass spectrometry was used to discern gypenosides and its possible metabolites in rat samples. As a result, after oral administration, a total of seven metabolites of G. pentaphyllum extract were assigned, two from the rat serum and seven both from the rat urine and faeces. As metabolites of G. pentaphyllum extract, all of them have never been reported before.

Estradiol synthesis and release in cultured female rat bone marrow stem cells.

Bone marrow stem cells (BMSCs) have the capacity to differentiate into mature cell types of multiple tissues. Thus, they represent an alternative source for organ-specific cell replacement therapy in degenerative diseases. In this study, we demonstrated that female rat BMSCs could differentiate into steroidogenic cells with the capacity for de novo synthesis of Estradiol-17 ß (E2) under high glucose culture conditions with or without retinoic acid (RA). The cultured BMSCs could express the mRNA and protein for P450arom, the enzyme responsible for estrogen biosynthesis. Moreover, radioimmunoassay revealed that BMSCs cultured in the present culture system produced and secreted significant amounts of testosterone, androstenedione, and E2. In addition, RA promoted E2 secretion but did not affect the levels of androgen. These results indicate that BMSCs can synthesize and release E2 and may contribute to autologous transplantation therapy for estrogen deficiency.

[STRA8 as a specific expression marker in postnatal male germ cells].

Stimulated by retinoic acid gene 8 (STRA8) is specifically expressed in mammalian germ cells before their transition from mitosis to meiosis. STRA8 expression is observed only in the postnatal testis. Single nucleotide polymorphisms but no mutations were identified in the coding or proximal promoter region of STRA8 in some gonadal dysgenesis patients. Studies on the teratocarcinoma cells and embryonic stem cells (ESC) transfected with STRA8-EGFP, a fusion construct harboring the promoter and coding region of the enhanced green fluorescence protein, have shown that the STRA8-EGFP positive cells may undergo meiosis, develop into sperm and generate live offspring mice. STRA8-EGFP positive cells derived from the bone marrow are able to differentiate into spermatogenic cells, but arrest in the premeiotic stage, and those from the adult mouse testis, when cultured in ESC culture conditions, may acquire ESC properties, pluripotency and redifferentiation capacity and act as a new stem cell source for tissue regeneration. The presence of oocytes renewed in postnatal mouse ovaries calls in question the absence of STRA8 in postnatal mouse ovaries.

[Application of fluorescence microscopic imaging technique with self-ordered ring to residues detection of antibiotics].

Fluorescence microscopic self-ordered ring (SOR) technique with microwave heating is proposed for minocycline based on the capillary flow of solvent on the surface of hydrophobic glass slide and applied to residues detection of the antibiotics in milks of Inner Mongolia. In the reaction medium of hexahydropyridine containing polyvinyl alcohol-124 (PVA-124), a SOR of minocycline with the outer diameter of 1.54 mm and the ring belt width of 22.6 microm can be formed. When a 0.30 microL droplet of minocycline mixture was spotted on the solid surface, minocycline in the range of 4.2 x 10(-2)-1.8 x 10(-11) mol x ring(-1) (1.4 x 10(-6)-0.60 x 10(-5) mol x L(-1)) can be detected, the maximum fluorescence intensity was found to be proportional to the minocycline concentration, and the limit of detection can reach 4.2 x 10(-13) mol x ring(-1) (1.4 x 10(-7) mol x L(-1)) with three times of signal to noise ratio. With the present method, the contents of minocycline in milk samples of Inner Mongolia and minocycline hydrochloride capsule sample were satisfactorily determined with recoveries of 97.2%-103% and 99.4%-102%, respectively, and RSD lower than 1.2%, correspondingly. Therefore, a highly sensitive and selective analysis method for detecting content of trace drug contaminations was established. The method provided theory basis to quantificational analysis of the residues of the antibiotics in milks which was Inner Mongolia' preponderant resource products. It has important realistic meaning and broad application prospect.

[Stra8: a specifically expressed gene in premeiotic germ cells' transition from mitosis to meiosis].

Stra8 (stimulated by retinoic acid gene 8) is a specific expression gene in mammalian germ cells' transition from mitosis to meiosis. Stra8 expresses in an anterior to posterior wave from embryonic days 12.5 to 16.5 in germ cells of XX gonads in mice. Meiosis germ cells are observed 1 day after Stra8 expression, which begins in the testis after birth and triggers meiosis entry. Germ cell sex determination is directed by a difference in the timing of entry into meiosis in embryonic gonads. The position of retinoic acid production and CYP26b1 expression that metabolizes RA to an inactive form regulates Stra8 expression. Although cytoplasmic protein Stra8 is necessary for the entry of germ cells into meiotic prophase, the function of the protein remains unknown.

[Proto-oncogene c-src regulates the viability of rat spermatogonial stem cells in vitro through phosphorylated signal transducer and activator of transcription-3].

The present study aimed to investigate the effect of proto-oncogene c-src on the viability of rat spermatogonial stem cells from 9 day-old rat in vitro. MTT method was used to observe the viability of the spermatogonial stem cells treated with antisense c-src oligodeoxynucleotides (ODNs) in vitro; RT-PCR was utilized to observe the expression of c-src mRNA and Western blot was used to observe the protein expressions of pp60c-src and phosphorylated signal transducer and activator of transcription-3 (p-STAT3). Compared with that in control group, the viability of spermatogonial stem cells decreased by 8.1% (P<0.05) and the expression of c-src mRNA decreased significantly after treatment with 10 µmol/L antisense c-src ODNs for 12 h. Compared with that in the control group, the protein expressions of pp60c-src and p-STAT3 decreased by 33.8% and 45.3% (both P<0.01), respectively, in the spermatogonial stem cells after being transfected with antisense c-src ODNs. The results suggest that proto-oncogene c-src regulates the viability of rat spermatogonial stem cells through p-STAT3.

[Expression of proto-oncogenes and its role in spermatogenic cells].

This article reviews the specific expression of many proto-oncogenes during male germ cell development. The normal expression of proto-oncogenes plays an important role in the regulation of spermatogonial mitosis, spermatocyte meiosis as well as spermiogenesis and sperm maturation.

[Effects of c-jun on hCG-induced testosterone secretion of rat Leydig cells in vitro].

OBJECTIVE: To investigate the effects of c-jun on hCG-induced testosterone secretion in isolated rat Leydig cells by antisense oligodeoxynucleotides(ASODNs). METHODS: c-jun ASODNs were used to antagonise the effects of c-jun, hCG was used to induce the testosterone secretion of LC cultured in vitro and testosterone was measured by radioimmunoassay. RESULTS: The testosterone secretion of LC in vitro could be induced by hCG, which was a good model for the functional study of LC. c-jun ASODNs decreased the hCG-induced testosterone secretion of LC in a dose-dependent manner(P < 0.05). CONCLUSION: It is suggested that c-jun proto-oncogene enhances the testosterone secretion of LC.

[Advances in the research of spermatogonial stem cell].

In recent years, people have paid more attention to the spermatogonial stem cells that have the capacity for self renewal and multilineage differentiation and produce daughter cells that can expand and differentiate into spermatozoa under the adjustment of self genes and external signal. This article reviews recent advances in studies of enrichment and original selection of the spermatogonial stem cells. This review also summarizes some control factors in proliferation and transplantation techniques.