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Toxoplasma gondii (T. gondii) is an intracellular protozoan that can cause toxoplasmosis in all warm-blooded hosts. This study focused on the prevalence and genetic characterize of T. gondii in ducks from Fujian province, China. Genomic DNA was extracted from duck tissue samples (heart, liver, lung, and muscle). To assess the genetic diversity of the T. gondii isolates, it was determined by using multilocus polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology. A total of 586 ducks from 5 cities in Fujian province were tested, and 35 (6.0%) of which were found to be positive for the T. gondii B1 gene. Further genotyping of these positive samples at 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) using PCR-RFLP revealed that one tissue samples (heart samples from Fuzhou ducks) were identified as Type I (ToxoDB#10). This study is the first report on the prevalence and genetic characterization of T. gondii in ducks in Fujian province, and Type I (ToxoDB#10) is found in ducks in China for the first time. The findings document the genetic characterization of T. gondii in free-range ducks from Fujian Province, thereby enriching the understanding of T. gondii genetic diversity in China. Moreover, these results provide essential data support for further prospective studies and underscores the "One Health" concept, emphasizing the integral link among human, animal, and environmental health.
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Patos , Doenças das Aves Domésticas , Toxoplasma , Toxoplasmose Animal , Animais , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/parasitologia , China/epidemiologia , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/epidemiologia , Prevalência , Polimorfismo de Fragmento de Restrição , Saúde Pública , Reação em Cadeia da Polimerase/veterinária , Genótipo , Variação GenéticaRESUMO
Toxoplasma gondii is an important protozoan pathogen, which can cause severe diseases in the newborns and immunocompromised individuals. Developing an effective vaccine against Toxoplasma infection is a critically important global health priority. Immunofluorescence staining analysis revealed that TgSAG2 and TgSRS2 are membrane associated and displayed on the surface of the parasite. Immunizations with pBud-SAG2, pBud-SRS2 and pBud-SAG2-SRS2 DNA vaccines significantly increased the production of specific IgG antibodies. Immunization with pBud-SAG2-SRS2 elicited cellular immune response with higher concentrations of IFN-γ and IL-4 compared to the control group. Antigen-specific lymphocyte proliferations in the pBud-SRS2 and pBud-SAG2-SRS2 groups were significantly higher compared to that in the control group. Furthermore, 30 % of mice immunized with pBud-SAG2-SRS2 survived after the challenge infection with virulent T. gondii RH tachyzoites. This study revealed that immunization with pBud-SAG2-SRS2 induced potent immune responses, and has the potential as a promising vaccine candidate for the control of T. gondii infection.
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Anticorpos Antiprotozoários , Antígenos de Protozoários , Imunoglobulina G , Proteínas de Protozoários , Vacinas Protozoárias , Toxoplasma , Toxoplasmose Animal , Vacinas de DNA , Animais , Vacinas de DNA/imunologia , Vacinas de DNA/genética , Vacinas de DNA/administração & dosagem , Antígenos de Protozoários/imunologia , Antígenos de Protozoários/genética , Proteínas de Protozoários/imunologia , Proteínas de Protozoários/genética , Toxoplasma/imunologia , Toxoplasma/genética , Anticorpos Antiprotozoários/sangue , Vacinas Protozoárias/imunologia , Vacinas Protozoárias/administração & dosagem , Vacinas Protozoárias/genética , Camundongos , Imunoglobulina G/sangue , Feminino , Toxoplasmose Animal/prevenção & controle , Toxoplasmose Animal/imunologia , Camundongos Endogâmicos BALB C , Interferon gama/imunologia , Modelos Animais de Doenças , Proliferação de Células , Interleucina-4/imunologia , Análise de SobrevidaRESUMO
N-acetylserotonin (NAS) is a chemical intermediate in melatonin biosynthesis. NAS and its derivative N-(2-(5-hydroxy-1H-indol-3-yl) ethyl)-2-oxopiperidine-3-carboxamide (HIOC) are potential therapeutic agents for traumatic brain injury, autoimmune encephalomyelitis, hypoxic-ischemic encephalopathy, and other diseases. Evidence shows that NAS and its derivative HIOC have neuroprotective properties, and can exert neuroprotective effects by inhibiting oxidative stress, anti-apoptosis, regulating autophagy dysfunction, and anti-inflammatory. In this review, we discussed the neuroprotective effects and related mechanisms of NAS and its derivative HIOC to provide a reference for follow-up research and applications.
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Encefalomielite Autoimune Experimental , Melatonina , Fármacos Neuroprotetores , Animais , Humanos , Fármacos Neuroprotetores/farmacologia , Serotonina/farmacologia , Neuroproteção , Melatonina/farmacologiaRESUMO
Toxoplasma gondii is an obligate intracellular protozoan parasite which seriously threatens the health of domestic animals and humans. Long non-coding RNAs (lncRNAs) are non-protein-coding transcripts greater than 200 nucleotides, which are widely involved in transcriptional and epigenetic regulations. However, little is known about the roles of host lncRNAs in the response to T. gondii infections. In this study, using Illumina sequencing technology, we analyzed the expression profiles of mRNAs and lncRNAs in BALB/c mouse brain following infection by T. gondii PRU strain (type II genotype) cysts. The identified differentially expressed (DE) RNAs were subjected to bioinformatics analysis. A total of 2,090 annotated lncRNAs along with 3,577 novel lncRNAs were identified. In the acutely infected mouse brain, a total of 330 mRNAs and 19 lncRNAs were dys-regulated, whereas 136 DE mRNAs and 9 DE lncRNAs were identified in chronically infected mouse brain. GO analysis revealed that these DE mRNAs identified at acute infection stage were involved in immune response, whereas DE mRNAs found at chronic infection stage were mostly enriched in response to protozoan. KEGG analysis showed that DE mRNAs were significantly enriched in disease related pathways. In addition, the putative mRNA-lncRNA co-expression network was constructed, and several hub regulatory RNAs were identified based on the transcriptome data. This study firstly characterized the co-expression profile of mRNAs and lncRNAs in mouse brain infected with T. gondii and provided a framework for further studies of the roles of lncRNAs in host neuropathology during toxoplasmosis progression.
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RNA Longo não Codificante , Toxoplasma , Toxoplasmose , Humanos , Camundongos , Animais , RNA Longo não Codificante/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Toxoplasmose/genética , Camundongos Endogâmicos BALB C , Encéfalo/metabolismo , Perfilação da Expressão GênicaRESUMO
Spinal cord injury (SCI) is a highly disabling disorder for which few effective treatments are available. Grape seed proanthocyanidins (GSPs) are polyphenolic compounds with various biological activities. In our preliminary experiment, GSP promoted functional recovery in rats with SCI, but the mechanism remains unclear. Therefore, we explored the protective effects of GSP on SCI and its possible underlying mechanisms. We found that GSP promoted locomotor recovery, reduced neuronal apoptosis, increased neuronal preservation, and regulated microglial polarisation in vivo. We also performed in vitro studies to verify the effects of GSP on neuronal protection and microglial polarisation and their potential mechanisms. We found that GSP regulated microglial polarisation and inhibited apoptosis in PC12 cells induced by M1-BV2 cells through the Toll-like receptor 4- (TLR4-) mediated nuclear factor kappa B (NF-κB) and phosphatidylinositol 3-kinase/serine threonine kinase (PI3K/AKT) signaling pathways. This suggests that GSP regulates microglial polarisation and prevents neuronal apoptosis, possibly by the TLR4-mediated NF-κB and PI3K/AKT signaling pathways.
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Fármacos Neuroprotetores , Traumatismos da Medula Espinal , Animais , Extrato de Sementes de Uva , Microglia/metabolismo , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Fosfatidilinositol 3-Quinases/metabolismo , Proantocianidinas , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Traumatismos da Medula Espinal/tratamento farmacológico , Traumatismos da Medula Espinal/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
BACKGROUND: The protozoan parasite Toxoplasma gondii is a major concern for human and animal health. Although the metabolic understanding of toxoplasmosis has increased in recent years, the analysis of metabolic alterations through noninvasive methodologies in biofluids remains limited. METHODS: Here, we applied liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics and multivariate statistical analysis to analyze BALB/c mouse urine collected from acutely infected, chronically infected and control subjects. RESULTS: In total, we identified 2065 and 1409 metabolites in the positive electrospray ionization (ESI +) mode and ESI - mode, respectively. Metabolomic patterns generated from principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) score plots clearly separated T. gondii-infected from uninfected urine samples. Metabolites with altered levels in urine from T. gondii-infected mice revealed changes in pathways related to amino acid metabolism, fatty acid metabolism, and nicotinate and nicotinamide metabolism. CONCLUSIONS: This is the first study to our knowledge on urine metabolic profiling of BALB/c mouse with T. gondii infection. The urine metabolome of infected mouse is distinctive and has value in the understanding of Toxoplasmosis pathogenesis and improvement of treatment.
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Toxoplasma , Toxoplasmose , Animais , Cromatografia Líquida , Humanos , Metaboloma , Metabolômica/métodos , Camundongos , Camundongos Endogâmicos BALB C , Espectrometria de Massas em Tandem , Toxoplasmose/parasitologiaRESUMO
BACKGROUND: Low back pain is a common symptom of intervertebral disc degeneration (IDD), which seriously affects the quality of life of patients. The abnormal apoptosis and senescence of nucleus pulposus (NP) cells play important roles in the pathogenesis of IDD. Proanthocyanidins (PACs) are polyphenolic compounds with anti-apoptosis and anti-aging effects. However, their functions in NP cells are not yet clear. Therefore, this study was performed to explore the effects of PACs on NP cell apoptosis and aging and the underlying mechanisms of action. METHODS: Cell viability was evaluated by cell counting kit-8 (CCK-8) assay. The apoptosis rate was determined TUNEL assays. Levels of apoptosis-associated molecules (Bcl-2, Bax, C-caspase-3 and Caspase-9) were evaluated via western blot. The senescence was observed through SA-ß-gal staining and western blotting analysis was performed to observe the expression of senescence-related molecules (p-P53, P53, P21 and P16). RESULTS: Pretreatment with PACs exhibited protective effects against IL-1ß-induced NP cell apoptosis including apoptosis rate, expressions of proapoptosis and antiapoptosis related genes and protein. PACs could also alleviate the increase of p-p53, P21, and P16 in IL-1ß-treated NP cells. SA-ß-gal staining showed that IL-1ß-induced senescence of NP cells was prevented by PACs pertreatment. In addition, PACs activated PI3K/Akt pathway in IL-1ß-stimulated NP cells. However, these protected effects were inhibited after LY294002 treatment. CONCLUSION: The results of the present study showed that PACs inhibit IL-1ß-induced apoptosis and aging of NP cells by activating the PI3K/Akt pathway, and suggested that PACs have therapeutic potential for IDD.
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Degeneração do Disco Intervertebral , Disco Intervertebral , Núcleo Pulposo , Proantocianidinas , Envelhecimento , Caspase 3/metabolismo , Caspase 9/metabolismo , Caspase 9/farmacologia , Células Cultivadas , Humanos , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/patologia , Núcleo Pulposo/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Proantocianidinas/metabolismo , Proantocianidinas/farmacologia , Proantocianidinas/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Qualidade de Vida , Proteína Supressora de Tumor p53/metabolismo , Proteína Supressora de Tumor p53/farmacologia , Proteína Supressora de Tumor p53/uso terapêutico , Proteína X Associada a bcl-2/metabolismo , Proteína X Associada a bcl-2/farmacologiaRESUMO
Background: Tripartite motif containing 46 was initially identified as the oncogene in several human tumors. However, the clinical value and potential functions of tripartite motif containing 46 (TRIM46) in clear cell renal cell carcinoma (ccRCC) remained largely unclear. Methods: The expressing patterns, clinical involvement, and prognostic values of TRIM46 were analyzed using the data obtained from TCGA and GEO databases. A nomogram was constructed to examine the outcome of patients with ccRCC. We estimated the association between TRIM46 with tumor immunity in ccRCC. Results: Tripartite motif containing 46 was highly expressed in ccRCC, and its upregulation revealed an unfavorable prognosis. A nomogram based on TRIM46 expressions and other independent prognostic factors could robustly predict the overall survival of tumor patients. TRIM46 has a strong positive correlation with NUMBL, CACNB1, THBS3, ROBO3, MAP3K12, ANKRD13D, PIF1, PRELID3A, ANKRD13B, and PCNX2. Mechanically, TRIM46 displayed regulatory functions in ccRCC progression via several tumor-associated pathways. Besides, we observed that TRIM46 was distinctly related to tumor immunity in ccRCC. Conclusions: Our findings provide a novel tumor promotive role regarding TRIM46 function in the malignant progression of ccRCC.
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Spinal cord injury (SCI) is a severe CNS injury that results in abnormalities in, or loss of, motor, sensory and autonomic nervous function. miRNAs belong to a new class of noncoding RNA that regulates the production of proteins and biological function of cells by silencing translation or interfering with the expression of target mRNAs. Following SCI, miRNAs related to oxidative stress, inflammation, autophagy, apoptosis and many other secondary injuries are differentially expressed, and these miRNAs play an important role in the progression of secondary injuries after SCI. The purpose of this review is to elucidate the differential expression and functional roles of miRNAs after SCI, thus providing references for further research on miRNAs in SCI.
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MicroRNAs , Traumatismos da Medula Espinal , Apoptose , Humanos , MicroRNAs/genética , RNA Mensageiro , Medula Espinal , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/terapiaRESUMO
BACKGROUND: Degenerative lumber spondylolisthesis (DLS) is a common orthopedic condition, described as a condition that compared with the lower vertebra, the superior vertebra slides forward or backward in the sagittal plane without accompanying isthmic spondylolisthesis. Information pertaining to different types of double-level DLS is scarce. This study aims to analyze parameters of patients with different types of double-level DLS to provide a reference for guiding surgical treatment and restoring sagittal balance of patients with DLS. METHODS: From January 2014 to January 2020, records of patients with double-level DLS were retrospectively reviewed. Patients with double-level DLS were divided into 3 types: anterior, posterior, and combined; the anterior and combined types were studied. The sagittal spinopelvic parameters included C7 tilt, maximal thoracic kyphosis, maximal lumbar lordosis (LLmax), pelvic incidence (PI), pelvic tilt (PT), and sacral slope (SS). After descriptive analysis, demographic and radiographic data were compared. RESULTS: Forty and 18 patients were included in the anterior and combined type groups, respectively. Both groups had different levels of chronic low back pain, but the incidence of radiating leg pain and neurogenic claudication was significantly higher in the anterior type. Oswestry Disability Index and visual analog scale low back scores were also higher in the anterior type. In the anterior type, C7 tilt (7.14 ± 2.15 vs. 5.41 ± 2.28, P = 0.007), LLmax (50.02 ± 14.76 vs. 36.96 ± 14.56, P = 0.003), PI (68.28 ± 9.16 vs. 55.53 ± 14.19, P < 0.001), PT (28.68 ± 7.31 vs. 19.38 ± 4.70, P < 0.001), and PT/PI (42.45 ± 11.22 vs. 36.04 ± 9.87, P = 0.041) were significantly higher. In the anterior type, PI correlated positively with LLmax (r = 0.59) and SS (r = 0.71). LLmax and SS (r = 0.65) had a positive correlation. PT/PI and SS (r = -0.77) had a negative correlation. In the combined type, PI correlated positively with LLmax (r = 0.61) and SS (r = 0.88), and PT/PI correlated negatively with SS (r = -0.81). CONCLUSIONS: In patients with double-level DLS, the sagittal spinopelvic parameters differed between the anterior and combined types. Overall, spinal surgeons should focus on correcting sagittal deformities, relieving postoperative clinical symptoms, and improving quality of life during fusion surgery.
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Degeneração do Disco Intervertebral/patologia , Vértebras Lombares/patologia , Espondilolistese/patologia , Idoso , Avaliação da Deficiência , Feminino , Humanos , Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/cirurgia , Cifose/patologia , Lordose/patologia , Dor Lombar/etiologia , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/cirurgia , Masculino , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos , Pelve/patologia , Estudos Retrospectivos , Fusão Vertebral , Espondilolistese/diagnóstico por imagem , Espondilolistese/cirurgiaRESUMO
Background: Glioblastoma (GBM) is one of the most malignant and aggressive primary brain tumors. The incurability of glioblastoma is heavily influenced by the glioma microenvironment. FTY720, a potent immunosuppressant, has been reported to exert anti-tumor effects in glioblastoma. However, the impact of FTY720 on the glioma microenvironment remains unclear. Methods: We examined the effects of FTY720 on the distribution and polarization of glioma-associated microglia and macrophages (GAMs) in glioma-bearing rats using immunofluorescence staining. qRT-PCR and Western blotting were used to detect the expressions of CXCR4 and MAPK pathway-related signal molecules on microglia in the coculture system. The levels of inflammatory factors were tested via ELISA. Wound healing assay and Matrigel invasion assay were used to determine the migration and invasion of C6 glioma cells. Results: We discovered that FTY720 could inhibit the growth, migration, and invasion of glioma by targeting GAMs to impede their effect on glioma cells. Simultaneously, FTY720 could block the chemoattraction of GAMs by inhibiting MAPK-mediated secretion of IL-6 through increased internalization of CXCR4. Moreover, microglia and macrophages are polarized from pro-glioma to an anti-tumor phenotype. Conclusion: These results provide novel insights into the inhibitory effects of FTY720 on glioma by targeting GAMs-glioma interaction in the tumor microenvironment.
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Antineoplásicos/administração & dosagem , Cloridrato de Fingolimode/administração & dosagem , Glioblastoma/tratamento farmacológico , Glioblastoma/metabolismo , Microglia/efeitos dos fármacos , Receptores CXCR4/metabolismo , Microambiente Tumoral/efeitos dos fármacos , Aloenxertos , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Polaridade Celular/efeitos dos fármacos , Modelos Animais de Doenças , Glioblastoma/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Microglia/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Receptores CXCR4/genética , Transdução de Sinais/efeitos dos fármacosRESUMO
Microbial oxidation is the only biological sink of atmospheric methane (CH4). It is essential to understand the variation of CH4 fluxes among different grassland use types for developing low-emission management system. Here, we measured the CH4 flux and the soil methane-oxidizing bacteria abundance in a typical steppe under grazing, mowing and fencing management in central Inner Mongolia, with the aims to determine the effects of these grassland use types on CH4 flux, and to test the hypothesis that pmoA functional gene abundance regulates CH4 fluxes. The measurements were conducted on the experimental grassland that had experienced four grassland use treatments over five years. The treatments were whole growing season grazing from May to September (T1), spring and summer grazing (twice in May and July)(T2), autumn mowing (T3) and enclosure (T0). We measured CH4 flux using static chamber method, and quantified the abundance of pmoA functional genes using molecular techniques. Moreover, we measured plant biomass and soil physicochemical properties. The results showed that moderate grazing significantly enhanced CH4 uptake rate and the methane-oxidizing bacteria abundance (i.e., the pmoA gene copy number per gram of dry soil). The pmoA gene copy number ranged from 6.9×104 to 3.9×105 per gram of dry soil in growing season. The CH4 uptake rate was (68.21±3.01) µg·m-2·h-1 under T1, which was 22.1%, 37.5% and 30.9% higher than that under T2, T3 or T0 , respectively. The CH4 uptake rate was positively correlated with abundance of CH4 oxidizing bacteria and soil sand content, but negatively correlated with soil silt content, soil moisture, NH4+-N and NO3--N content, and plant biomass. These results suggested that the steppe ecosystem is a CH4 sink under all land-use types in central Inner Mongolia, and that moderate grazing would enhance methane-oxidizing bacteria abundance and CH4 uptake by improving soil sand content, reducing soil mineral nitrogen content and plant production in the typical steppe ecosystem. These results were of significance for the development of low-emission grassland management system.
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Ecossistema , Metano/análise , Methylococcaceae/fisiologia , China , SoloRESUMO
Women are believed to be more vulnerable to develop depressive symptoms during the perimenopause compared to postmenopause. The traditional bilateral ovariectomy and chronic mild stress (CMS) stimulation animal model produces a postmenopausal depressive-like state but the transition from perimenopausal period to postmenopausal period was ignored. Thus we establish a novel animal model in which the mice were stimulated by CMS for three months and removed the ovaries by two-step operation, and then evaluate whether this novel model could be much better for preclinical study used as a peri/postmenopause depressive model. The present study systemically evaluated the changes induced by two-step ovariectomy plus CMS in the mice. The depression-like behaviors, the levels of corticosterone, estrogen, pro-inflammatory factors, neurotransmitters, as well as brain-derived neurotrophic factor were determined; the changes of estrogen receptors, serotonin receptors, uterine weight and bone microarchitecture were also observed. The results show that the behaviors and biochemical indexes of mice changed gradually over time. Our study suggests that this two-step ovariectomy operation plus CMS successfully establishes a more reasonable peri/postmenopausal depression animal model which effectively simulates the clinical symptoms of peri/postmenopausal depressive women.
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Vascular endothelial growth factor (VEGF) is a dimeric glycoprotein that induces proliferation and migration of vascular endothelial cells as well as regulation of capillary formation around hair follicles which affects the growth and development of hair follicles. cgVEGF164 is a major splice variant of the cashmere goat VEGF-A gene, but its regulation on hair follicles is rarely known. In order to investigate the role of cgVEGF164 on the growth of murine hair follicles, we produced keratin 14 promoter-driven cgVEGF164 transgenic mice via pronuclear microinjection. Firstly, the diameter and density of hair follicles of transgenic mice were compared with non-transgenic control mice in paraffin sections stained by hematoxylin-eosin (H&E). Then, protein expression levels and the phosphorylation of ERK1/2, AKT1 and LEF1 were examined by Western blot. There are five positive individuals among the neonatal mice (positive rate is 8.5%). Compared with non-transgenic control mice, the diameter and density of hair follicles in transgenic mice are both obviously increased. The expression levels of P-ERK1/2/ERK1/2, P-AKT1/AKT1 and P-LEF1/LEF1 are significantly higher in transgenic mice than those in non-transgenic control mice. Based on these results, we conclude that cgVEGF164 as a growth factor can improve the growth of hair follicles which might be mediated by increasing the levels of ERK1/2, AKT1, and LEF1 protein phosphorylation.
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Folículo Piloso/crescimento & desenvolvimento , Isoformas de Proteínas/farmacologia , Fator A de Crescimento do Endotélio Vascular/farmacologia , Animais , Cabras , Camundongos , Camundongos Transgênicos , FosforilaçãoRESUMO
Increased cashmere yield and improved quality are some goals of cashmere goat breeding. Thymosin beta-4 (Tß4) plays a key role in the growth and development of hair follicles. For the past ten years, we have evaluated the role of Tß4 by establishing a flock of 15 cashmere goats that specifically overexpress the Tß4 gene in the hair follicles. These Tß4 overexpression (Tß4-OE) cashmere goats had more secondary hair follicles than the WT goats and produced more cashmere. Meanwhile, combined analysis of the skin transcriptome and proteome in cashmere goats suggested that Tß4 may affect hair growth by interacting with keratin type II cytoskeletal 4 epidermal (KRT4) to mediate the extracellular signal-regulated protein kinase (ERK) signaling pathway, thereby promoting the development of secondary hair follicles, and consequently, increasing cashmere yield. Thus, the specific overexpression of Tß4 in the hair follicles of cashmere goats effectively increased the cashmere yield.
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Traditional bulk polymerization imprinted technology and existing surface imprinted technology have some congenital defects. Therefore, it is necessary to design more efficient surface imprinted technology. In this paper, novel surface imprinting technology with higher imprinting efficiency is well designed. It fully realizes the synchronization of polymer crosslinking and template imprinting. Then the surface imprinted polymers (SIPs) are synthesized using metal ions as a template. The physicochemical characteristics of the SIPs are characterized by scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET) studies, Fourier transform infrared spectroscopy (FTIR) and elemental analysis. The adsorption performances and recognition selectivity of the SIPs towards the template are investigated by a batch method. The experimental results show that the SIPs possess excellent adsorption ability and selectivity towards the template. The selectivity coefficients of the SIPs prepared in this study are higher than those of IIPs prepared by other imprinting methods. The adsorption process could be well described by the Lagergren-first-order model and Langmuir monolayer chemical adsorption. The SIPs have good chemical stability and reusability. Consecutive adsorption-desorption experiments show that the exhausted SIPs could be effectively regenerated, and the regenerated SIPs could be reused without a significant reduction in adsorption capacity or selectivity coefficient.
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Microglia-mediated neuroinflammation plays a dual role in various brain diseases due to distinct microglial phenotypes, including deleterious M1 and neuroprotective M2. There is growing evidence that the peroxisome proliferator-activated receptor γ (PPARγ) agonist rosiglitazone prevents lipopolysaccharide (LPS)-induced microglial activation. Here, we observed that antagonizing PPARγ promoted LPS-stimulated changes in polarization from the M1 to the M2 phenotype in primary microglia. PPARγ antagonist T0070907 increased the expression of M2 markers, including CD206, IL-4, IGF-1, TGF-ß1, TGF-ß2, TGF-ß3, G-CSF, and GM-CSF, and reduced the expression of M1 markers, such as CD86, Cox-2, iNOS, IL-1ß, IL-6, TNF-α, IFN-γ, and CCL2, thereby inhibiting NFκB-IKKß activation. Moreover, antagonizing PPARγ promoted microglial autophagy, as indicated by the downregulation of P62 and the upregulation of Beclin1, Atg5, and LC3-II/LC3-I, thereby enhancing the formation of autophagosomes and their degradation by lysosomes in microglia. Furthermore, we found that an increase in LKB1-STRAD-MO25 complex formation enhances autophagy. The LKB1 inhibitor radicicol or knocking down LKB1 prevented autophagy improvement and the M1-to-M2 phenotype shift by T0070907. Simultaneously, we found that knocking down PPARγ in BV2 microglial cells also activated LKB1-AMPK signaling and inhibited NFκB-IKKß activation, which are similar to the effects of antagonizing PPARγ. Taken together, our findings demonstrate that antagonizing PPARγ promotes the M1-to-M2 phenotypic shift in LPS-induced microglia, which might be due to improved autophagy via the activation of the LKB1-AMPK signaling pathway.
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Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia/efeitos dos fármacos , Microglia/efeitos dos fármacos , PPAR gama/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/metabolismo , Transdução de Sinais/efeitos dos fármacos , Quinases Proteína-Quinases Ativadas por AMP , Animais , Benzamidas/farmacologia , Células Cultivadas , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Microglia/metabolismo , PPAR gama/metabolismo , Piridinas/farmacologia , Ratos , Ratos Sprague-Dawley , Rosiglitazona/farmacologiaRESUMO
Fibroblast growth factors (FGFs) are multifunctional signal molecules between cells, regulating the various physiological functions of the organism. FGF21 is a regulatory factor of the FGF family and has been postulated to play important roles in hair follicle development and hair follicle growth cycle. To evaluate the roles of FGF21, we had established a FGF21 knockout mouse model, using the CRISPR/Cas9 technology. We had constructed a FGF21 targeting vector and microinjected it with Cas9 mRNA and gRNA into fertilized ova of FVB mice. The gRNA was designed to target the exon 1 of the endogenous mouse FGF21 gene. Three lines of Fgf21 -/- mice were obtained from these experiments, and confirmed to harbor Fgf21 -/- genotypes and null expression phenotype, using DNA sequencing, qRT-PCR and Western blotting. FGF21 mRNA and FGF21 protein were not detected in tissues of these Fgf21 -/- mice. Depilation and histochemistry analyses showed that the Fgf21 -/- mice had lower body weight, slower hair regrowth and poorer hair quantities and smaller hair follicles diameters, as compared to WT mice. The Fgf21 -/- mice reported here could provide a useful genetic model for future studies of FGF21 functions in hair follicle development and hair follicle growth cycle.
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Sistemas CRISPR-Cas , Fatores de Crescimento de Fibroblastos/genética , Animais , Feminino , Fatores de Crescimento de Fibroblastos/fisiologia , Folículo Piloso/crescimento & desenvolvimento , Camundongos , Camundongos KnockoutRESUMO
Therapeutic hypothermia is a promising strategy for acute cerebral ischemia via physical or pharmacological methods. In this study, we pharmacologically induced hypothermia on Sprague Dawley rats by intraperitoneally injecting PD149163. We found that mild hypothermia was induced by PD149163 treatment without local cerebral blood flow (LCBF) alteration. To evaluate the neuroprotective effects of PD149163, TTC staining, HE staining and Nissl's staining were performed in our study. We found that PD149163 could prevent neuronal damage, and inhibit proliferation and activation of glial cells induced by ischemia. Simultaneously, we observed PD149163 ameliorated apoptosis characterized by down-regulated caspase-3 and Bax, but elevated Bcl-2. Moreover, PD149163 dramatically reduced JNK and AMPK/mTOR signaling pathway activation, and thereby inhibited autophagy by increased P62 expression, decreased the ratio of LC3-â ¡ to LC3-â and the expression of Beclin. Taken together, the present findings reveal the therapeutic effects of PD149163-induced hypothermia in brain ischemia, and provide a new strategy for stroke treatment.
Assuntos
Isquemia Encefálica/complicações , Hipotermia Induzida , Hipóxia Encefálica/etiologia , Hipóxia Encefálica/prevenção & controle , Fármacos Neuroprotetores , Oligopeptídeos/administração & dosagem , Oligopeptídeos/farmacologia , Doença Aguda , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Autofagia/genética , Proliferação de Células/efeitos dos fármacos , Injeções Intraperitoneais , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Neuroglia/citologia , Ratos Sprague-DawleyRESUMO
There is increasing interest in the association between depression and the development of metabolic diseases. Rosiglitazone, a therapeutic drug used to treat type 2 diabetes mellitus, has shown neuroprotective effects in patients with stroke and Alzheimer's disease. The present study was performed to evaluate the possible roles of rosiglitazone in in vivo (unpredictable chronic mild stress-induced depressive mouse model) and in vitro (corticosterone-induced cellular model) depressive models. The results showed that rosiglitazone reversed depressive behaviors in mice, as indicated by the forced swimming test and open field test. Rosiglitazone was also found to inhibit the inflammatory response, decrease corticosterone levels, and promote astrocyte proliferation and neuronal axon plasticity in the prefrontal cortex of mice. This series of in vivo and in vitro experiments showed that autophagy among neurons was inhibited in depressive models and that rosiglitazone promoted autophagy by upregulating LKB1, which exerted neuroprotective effects. Rosiglitazone was also found to activate the Akt/CREB pathway by increasing IGF-1R expression and IGF-1 protein levels, thereby playing an anti-apoptotic role in astrocytes. Rosiglitazone's autophagy promotion and neuroprotective effects were found to be reversed by the PPARγ antagonist T0070907 in primary neurons and by PPARγ knockdown in an N2a cell line. In conclusion, we found that rosiglitazone protects both neurons and astrocytes in in vivo and in vitro depressive models, thereby playing an anti-depressive role. These findings suggest that PPARγ could be a new target in the development of anti-depressive drugs.