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Two novel rod-shaped, strictly aerobic, non-motile and Gram-stain-negative bacterial strains, designated SDUM040013T and SDUM040014T, were isolated from kelp seedlings in Weihai, PR China. Cells of strain SDUM040013T were 0.3-0.4 µm wide and 0.8-1.8 µm long, catalase-positive and oxidase-positive. Growth of SDUM040013T was observed at 0-37 °C (optimum, 28-30 °C) and pH 5.5-9 (optimum, pH 8.0) and in the presence of 1-8â% (w/v) NaCl (optimum, 2â%). The DNA G+C content of strain SDUM040013T was 50.5â%. Strain SDUM040013T showed the highest 16S rRNA gene sequence similarity (97.1â%) to Gilvimarinus chinensis. Cells of strain SDUM040014T were 0.4-0.5 µm wide and 1.0-1.4 µm long, catalase-positive and oxidase-positive. Growth of SDUM040014T was observed at 4-40 °C (optimum, 28-30 °C) and pH 5.5-9 (optimum, pH 8.5) and in the presence of 0-8â% (w/v) NaCl (optimum, 2â%). The DNA G+C content of strain SDUM040014T was 56.5â%. Strain SDUM040014T showed the highest 16S rRNA gene sequence similarity (96.2%) to Gilvimarinus polysaccharolyticus. The isoprenoid quinone of both strains was Q-8 and the predominant fatty acids were summed feature 3 (C16â:â1 ω7c and/or C16â:â1 ω6c), summed feature 8 (C18â:â1 ω7c) and C16â:â0. Diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine were the major polar lipids. Given these phenotypic and chemotaxonomic properties, as well as phylogenetic data, strains SDUM040013T and SDUM040014T were considered to represent two novel species of the genus Gilvimarinus, for which the names Gilvimarinus gilvus sp. nov. and Gilvimarinus algae sp. nov. are proposed. The type strains are SDUM040013T (=KCTC 8123T=MCCC 1H01413T) and SDUM040014T (=KCTC 8124T=MCCC 1H01414T), respectively.
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Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano , Ácidos Grasos , Kelp , Filogenia , ARN Ribosómico 16S , Plantones , Análisis de Secuencia de ADN , ARN Ribosómico 16S/genética , Ácidos Grasos/química , China , ADN Bacteriano/genética , Kelp/microbiología , Plantones/microbiología , Ubiquinona/análogos & derivadosRESUMEN
An orange-pigmented, Gram-stain-negative, strictly aerobic, non-flagellated and rod-shaped bacterium, designated strain DF17T, was isolated from coastal sediment collected from Jingzi Wharf, Weihai, PR China. The optimal growth conditions were determined to be at 30 °C, pH 7.5, and in 3â% (w/v) NaCl. According to phylogenetic analysis of the 16S rRNA gene sequence, strain DF17T showed the highest sequence similarity of 96.9â% to Winogradskyella aquimaris KCTC 23502T. The DNA G+C content was 35.8âmol%, and the major fatty acids were iso-C15â:â1 G, iso-C15â:â0, and iso-C17â:â0 3-OH. The major polar lipids were two aminoglycolipids, one phosphatidylethanolamine and four unidentified lipids. The predominant respiratory quinone was menaquinone-6 (MK-6). The average nucleotide identity, digital DNA-DNA hybridization, and amino acid identity values between strain DF17T and other Winogradskyella species were below the species delineation thresholds of 69.35-72.95â%, 16.9-19.6â% and 71.25-78.93â%, respectively. On the basis of its phenotypic, genetic and physiological characteristics, strain DF17T is suggested to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella pelagia sp. nov. is proposed. The type strain is DF17T (MCCC 1H00456T=KCTC 82421T).
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Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano , Ácidos Grasos , Sedimentos Geológicos , Hibridación de Ácido Nucleico , Filogenia , ARN Ribosómico 16S , Análisis de Secuencia de ADN , Vitamina K 2 , ARN Ribosómico 16S/genética , Sedimentos Geológicos/microbiología , Ácidos Grasos/química , ADN Bacteriano/genética , Vitamina K 2/análogos & derivados , Vitamina K 2/análisis , China , Agua de Mar/microbiología , Datos de Secuencia Molecular , FosfatidiletanolaminasRESUMEN
PURPOSE: To evaluate the impact of two different parameters (body position and distension medium) on the rectal sensory test in patients with functional constipation and provide data support for the development of standardized operating procedures in clinical practice. METHODS: Based on a single-center process of the rectal sensory test, 39 patients with functional constipation were recruited for rectal sensory test under different body positions and distension mediums. RESULTS: Among the items of the Constipation Scoring System, the score of frequency of bowel movements showed a negative correlation with the first constant sensation volume (r = -0.323, P = 0.045). Conversely, the score of painful evacuation effort showed a positive correlation with the desire to defecate volume (r = 0.343, P = 0.033). There was a statistically significant difference in the first constant sensation volume (when the distension medium was gas) measured in different body positions (left lateral position, sitting position, squatting position), and the data measured in the squatting position were significantly higher than those in left lateral position (P < 0.05). In terms of research on distension medium, it was found that the first constant sensation volume measured in the squatting position (when the distension medium was water) was significantly lower than that of gas (P < 0.05). CONCLUSION: For patients with functional constipation, there are differences in the results of rectal sensory tests between body positions and distension mediums. When conducting multicenter studies, it is necessary to unify the standard operating procedure (SOP) for operational details to ensure consistency and reliability of the test results.
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Estreñimiento , Posicionamiento del Paciente , Recto , Humanos , Estreñimiento/fisiopatología , Estreñimiento/diagnóstico , Femenino , Recto/fisiopatología , Masculino , Adulto , Persona de Mediana Edad , Posicionamiento del Paciente/métodos , Defecación/fisiología , Sensación/fisiología , Anciano , Adulto JovenRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Scutellaria baicalensis Georgi (Scutellariae Radix, SR) and Coptis chinensis Franch (Coptidis Rhizoma, CR) is a classic herbal pair used in many Traditional Chinese Medicine formulations in the treatment of hyperlipidemia (HLP). As effective ingredients of the drug pair, the effects and mechanisms of berberine and baicalin in the treatment of HLP in the form of components compatibility are still unclear. AIM OF THE STUDY: To explore the mechanism of the components compatibility of SR and CR in the treatment of HLP. MATERIALS AND METHODS: The HLP model was established by a high-fat diet. Serum biochemical indexes were detected. Transcriptomics and metabolomics were detected. RT-PCR and Western Blot were used to analyze the effect of RA on the expression of the Cyp4a family during the treatment of HLP. RESULTS: Berberine-baicalin (RA) has a good effect in the treatment of HLP. RA can significantly reduce the body weight and liver weight of HLP, reduce the levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), and increase the level of high-density lipoprotein (HDL-C). Through transcriptomic analysis, RA significantly reversed the gene expression of Cyp4a10, Cyp4a12 b, Cyp4a31, and Cyp4a32 in cytochrome P450 family 4 subfamily a (Cyp4a) which related to fatty acid degradation in the liver of HLP mice. The results of fatty acid detection showed that RA could significantly regulate heptanoic acid, EPA, adrenic acid, DH-γ-linolenic acid, and DPA in the cecum of HLP mice. The Cyp4a family genes regulated by RA are closely related to a variety of fatty acids regulated by RA. RT-PCR confirmed that RA could regulate Cyp4a mRNA expression in HLP mice. WB also showed that RA can regulate the protein expression level of Cyp4a. CONCLUSION: The components compatibility of SR and CR can effectively improve the blood lipid level of HLP mice, its mechanism may be related to regulating Cyp4a gene expression and affecting fatty acid degradation, regulating the level of fatty acid metabolism in the body.
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Berberina , Coptis chinensis , Dieta Alta en Grasa , Medicamentos Herbarios Chinos , Hiperlipidemias , Scutellaria baicalensis , Animales , Hiperlipidemias/tratamiento farmacológico , Scutellaria baicalensis/química , Masculino , Berberina/farmacología , Berberina/uso terapéutico , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/química , Ratones , Flavonoides/farmacología , Ratones Endogámicos C57BL , Familia 4 del Citocromo P450/genética , Familia 4 del Citocromo P450/metabolismo , Coptis/química , Hipolipemiantes/farmacología , Hígado/efectos de los fármacos , Hígado/metabolismo , Lípidos/sangre , Modelos Animales de Enfermedad , RizomaRESUMEN
A Gram-stain-negative, rod-shaped, non-gliding, non-flagellated, yellow, facultatively aerobic bacterial strain, designated as W260T, was isolated from marine sediment of Xiaoshi Island, Weihai, PR China. The cells of W260T were 0.3-0.5 µm wide and 1.5-2.0 µm long. Strain W260T grows optimally at a temperature of 33â°C (range, 15-37â°C), pH 8 (range, pH 6.5-9.5) and witha NaCl concentration of 3.0â% (w/v; range, 1-8â%). It has the highest sequence similarity to Thiohalobacter thiocyanaticus DSM 21152T (91.7â%), followed by Wenzhouxiangella marina MCCC 1K00261T (91.4â%) and Thiohalospira alkaliphila DSM 17116T (90.7â%). The similarity between strain W260T and the species Thiohalophilus thiocyanatoxydans DSM 16326T was 89.4â%. Genome sequencing revealed a genome size of 3â430â000 bp and a DNA G+C content of 64.5âmol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain W260T and W. marina MCCC 1K00261T were 69.6 and 16.1-20.6â%, respectively. The predominant quinone was ubiquitin-8, and the major fatty acids were iso-C14â:â0 and iso-C16â:â0. The polar lipids consisted of phosphatidylethanolamine, phospholipid, phosphatidylglycerol, diphosphatidylglycerol and four unidentified lipids. Based on phenotypic, phylogenetic and chemotaxonomic information, it was determined that strain W260T represents a novel genus and species and it was given the name Marinihelvus fidelis sp. nov. The type strain is W260T (=MCCC 1H00471T=KCTC 92639T).
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Ácidos Grasos , Sedimentos Geológicos , Ácidos Grasos/química , Filogenia , Análisis de Secuencia de ADN , Composición de Base , Técnicas de Tipificación Bacteriana , ARN Ribosómico 16S/genética , ADN Bacteriano/genética , Sedimentos Geológicos/microbiología , Fosfolípidos/química , GenómicaRESUMEN
Pseudorabies (PR) and classical swine fever (CSF) are economically important infectious diseases in pigs. Most pig farms in China are vaccinated against these two diseases. Gene-deleted pseudorabies virus (PRV) can be used to develop promising and economical multivalent live attenuated viral vector vaccines. It has been reported that recombinant PRV can express a truncated E2 protein (1-338 aa), but it has not been reported that recombinant PRV can express a full-length E2 protein. We constructed nine groups of E2 proteins with different expression forms and found that the E2 protein could be expressed in vitro only when the transmembrane region of E2 was removed and the signal peptide was added. Analysis of the transmembrane region of E2 revealed that the high hydrophobicity of the E2 transmembrane region was the main reason for its inability to express. By mutating an amino acid to reduce the hydrophobicity of the transmembrane region, it was found that the full-length mutant of E2 (E2FL-muta3 or E2FL-muta4) could be expressed. The expressed full-length mutant E2 could also localize to the cell membrane. Mice immunized with a PRV vector vaccine expressing E2FL-muta3 or E2FL-muta4 developed specific cellular immunity to the E2 protein and stimulated higher levels of E2 antibody than mice immunized with a PRV vector expressing truncated E2. After immunizing the rabbits, the lethal challenge by PRV-ZJ2013 and the febrile response elicited by CSFV were simultaneously prevented. These results suggest that rPRV-dTK/gE-E2FL-muta4 is a promising bivalent vaccine against CSFV and PRV infections.
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Virus de la Fiebre Porcina Clásica , Peste Porcina Clásica , Herpesvirus Suido 1 , Seudorrabia , Enfermedades de los Porcinos , Vacunas Virales , Animales , Porcinos , Ratones , Conejos , Herpesvirus Suido 1/genética , Virus de la Fiebre Porcina Clásica/genética , Aminoácidos/genética , Vacunas Virales/genética , Anticuerpos Antivirales , Inmunización , Seudorrabia/prevención & control , Mutación , Proteínas del Envoltorio Viral/genéticaRESUMEN
Pseudorabies (PR) and classical swine fever (CSF) are economically important infectious diseases of pigs. Most pig farms in China are immunized against these two diseases. Here, we describe a stabilized E2 protein as an immunogen inserted into the PRV genome as a bivalent live virus-vectored vaccine. The E2 protein has 48 variant sites, there are 2-5 candidate amino acids per variant site, and the relative energy contribution of each amino acid to E2 energy was calculated. Combined substitutions of amino acids at the neighbor variant site (neighbor substitution) were performed to obtain the E2 protein sequence with the lowest energy (stabilized E2). Multiple amino acid substitutions at 48 variant sites were performed, and the results were consistent with neighbor substitutions. The stabilized E2 sequence was obtained, and its energy decreased by 22 Rosetta Energy Units (REUs) compared with the original sequence. After the recombinant PRV expressing stabilized E2 of CSFV was constructed, the secretion efficiency of stabilized E2 was increased by 2.97 times, and the thermal stability was increased by 10.5 times. Immunization of mice resulted in a 2-fold increase in antibody production, and a balanced antibody level against subtype 1.1 and subtype 2.1d E2 was achieved. In rabbits immunized, the lethal challenge of PRV-ZJ and the fever response induced by CSFV could be prevented simultaneously. These findings suggest that rPRV-muta/287aaE2 is a promising bivalent vaccine against CSFV and PRV infections.
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Virus de la Fiebre Porcina Clásica , Peste Porcina Clásica , Herpesvirus Suido 1 , Seudorrabia , Vacunas Virales , Conejos , Animales , Porcinos , Ratones , Virus de la Fiebre Porcina Clásica/genética , Herpesvirus Suido 1/genética , Seudorrabia/prevención & control , Aminoácidos , Proteínas del Envoltorio Viral/genética , Anticuerpos AntiviralesRESUMEN
AeP-P-2, a pectic polysaccharide, was extracted from the fruit pod of okra. It composed of rhamnose (Rha), arabinose (Ara), glucose (Glc), galactose (Gal) and galacturonic acid (GalA) with the ratio of 4.75:2.01:1.00:4.91:7.24. The main structural feature of AeP-P-2 are 1,4-linked galacturonan units (homogalacturonan backbone) and (1 â 2) and (1 â 2,4) linked Rha (rhamnogalacturonan I region). And the other side chains contained â1)-linked Ara, (1 â 5)-linked Ara, (1 â 4)-linked Glc, (1 â 6)-linked Gal, (1 â 4)-linked Rha, (1 â 2,4)-linked Rha, â1)-linked Ara and â1)-linked Gal. When the concentration of AeP-P-2 was 3.2 mg/mL, the scavenging rates on DPPH·, ABTS, O2-· and ·OH reached to 61.88%, 87.10%, 52.17% and 60.32%, respectively. AeP-P-2 also could protect PC12 cells from the damage of H2O2 and reduce apoptosis caused by oxidative damage by decreasing the level of ROS. The findings indicated that okra was a functional vegetable and AeP-P-2 was worth studying and developing into antioxidant component.
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Abelmoschus , Abelmoschus/química , Animales , Antioxidantes/química , Antioxidantes/farmacología , Peróxido de Hidrógeno , Células PC12 , Polisacáridos/química , Polisacáridos/farmacología , RatasRESUMEN
Lung ischemia-reperfusion (IR) injury is induced by pulmonary artery occlusion and reperfusion. Lung IR injury commonly happens after weaning from extracorporeal circulation, lung transplantation, and pulmonary thromboendarterectomy; it is a lethal perioperative complication. A definite therapeutic intervention remains to be determined. It is known that the enzyme activity of angiotensin-converting enzyme 2 (ACE2) is critical in maintaining pulmonary vascular tone and epithelial integrity. In a noxious environment to the lungs, inactivation of ACE2 is mainly due to a disintegrin and metalloprotease 17 (ADAM17) protein-mediated ACE2 shedding. Thus, we assumed that protection of local ACE2 in the lung against ADAM17-mediated shedding would be a therapeutic target for lung IR injury. In this study, we established both in vivo and in vitro models to demonstrate that the damage degree of lung IR injury depends on the loss of ACE2 and ACE2 enzyme dysfunction in lung tissue. Treatment with ACE2 protectant diminazen aceturate (DIZE) maintained higher ACE2 enzyme activity and reduced angiotensin II, angiotensin type 1 receptor, and ADAM17 levels in the lung tissue. Concurrently, DIZE-inhibited oxidative stress and nitrosative stress via p38MAPK and NF-κB pathways consequently reduced release of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1ß. The underlying molecular mechanism of DIZE contributed to its protective effect against lung IR injury and resulted in the improvement of oxygenation index and ameliorating pulmonary pathological damage. We concluded that DIZE protects the lungs from IR injury via inhibition of ADAM17-mediated ACE2 shedding.
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Steam explosion (SE) was a friendly environmentally pretreatment method. In this study, the effect of steam explosion (SE) pretreatment on structure and α-glucosidase inhibitory activity of Ampelopsis grossedentata polysaccharides was evaluated. Two novel polysaccharides (AGP and AGP-SE) were extracted, isolated, purified and analyzed by NMR, FT-IR and methylation. The results indicated that AGP mainly consisted of Rha, Xyl, Glc, and Ara with a molecular weight of 2.74 × 103 kDa and AGP-SE mainly consisted of Man, Ara, and Gal with a molecular weight of 2.14 × 103 kDa. Furthermore, the backbone of AGP and AGP-SE were mainly composed of 5)-Araf-(1â, -Glcp-(1â, 6)-Glcp-(1â, 6)-Galp-(1â, 3,6)-Manp-(1â, and 2,3,6)-Glcp-(1â. Finally, we demonstrated that all polysaccharides exhibited obviously α-glucosidase inhibition activity and mixed type inhibition. AGP-SE had better α-glucosidase inhibition activity and the binding affinity KD on α-glucosidase by using Surface Plasmon Resonance (SPR) than AGP. Overall, SE pretreatment is an effective method for extracting polysaccharide and provides a new idea into the improvement of biological activity.
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Ampelopsis/química , Inhibidores de Glicósido Hidrolasas/farmacología , Polisacáridos/farmacología , alfa-Glucosidasas/metabolismo , Secuencia de Carbohidratos , Inhibidores de Glicósido Hidrolasas/química , Inhibidores de Glicósido Hidrolasas/aislamiento & purificación , Humanos , Metilación , Peso Molecular , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Espectroscopía Infrarroja por Transformada de Fourier , Vapor , Resonancia por Plasmón de SuperficieRESUMEN
BACKGROUND: Alzheimer's disease (AD) is a progressive neurodegenerative disease. One of the pathologies of AD is the accumulation of amyloid-ß (Aß) to form senile plaques, leading to a decline in cognitive ability and a lack of learning and memory. However, the cause leading to Aß aggregation is not well understood. Dendritic cell factor 1 (Dcf1) shows a high expression in the entorhinal cortex neurons and neurofibrillary tangles in AD patients. OBJECTIVE: Our goal is to investigate the effect of Dcf1 on Aß aggregation and memory deficits in AD development. METHODS: The mouse and Drosophila AD model were used to test the expression and aggregation of Aß, senile plaque formation, and pathological changes in cognitive behavior during dcf1 knockout and expression. We finally explored possible drug target effects through intracerebroventricular delivery of Dcf1 antibodies. RESULTS: Deletion of Dcf1 resulted in decreased Aß42 level and deposition, and rescued AMPA Receptor (GluA2) levels in the hippocampus of APP-PS1-AD mice. In Aß42 AD Drosophila, the expression of Dcf1 in Aß42 AD flies aggravated the formation and accumulation of senile plaques, significantly reduced its climbing ability and learning-memory. Data analysis from all 20 donors with and without AD patients aged between 80 and 90 indicated a high-level expression of Dcf1 in the temporal neocortex. Dcf1 contributed to Aß aggregation by UV spectroscopy assay. Intracerebroventricular delivery of Dcf1 antibodies in the hippocampus reduced the area of senile plaques and reversed learning and memory deficits in APP-PS1-AD mice. CONCLUSION: Dcf1 causes Aß-plaque accumulation, inhibiting dcf1 expression could potentially offer therapeutic avenues.
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Péptidos beta-Amiloides/metabolismo , Hipocampo/metabolismo , Proteínas de la Membrana/genética , Trastornos de la Memoria/genética , Proteínas del Tejido Nervioso/genética , Agregación Patológica de Proteínas/genética , Anciano de 80 o más Años , Precursor de Proteína beta-Amiloide/genética , Precursor de Proteína beta-Amiloide/metabolismo , Animales , Condicionamiento Clásico/fisiología , Drosophila melanogaster , Hipocampo/patología , Humanos , Aprendizaje/fisiología , Proteínas de la Membrana/metabolismo , Memoria/fisiología , Trastornos de la Memoria/metabolismo , Ratones , Ratones Noqueados , Ratones Transgénicos , Proteínas del Tejido Nervioso/metabolismo , Agregación Patológica de Proteínas/metabolismo , Agregación Patológica de Proteínas/patología , Receptores AMPA/metabolismoRESUMEN
Single herbs and Chinese patent medicine preparations often have bad taste, such as bitterness and astringency, which is one of the key factors affecting patients' medication compliance, and would affect the therapeutic effect and restrict the extensive application in clinical practice. Therefore, how to make use of taste masking techniques to improve the bad taste of traditional Chinese medicines has become an important project. Through the collection and summarization of Chinese and foreign journals and papers in recent years, this paper discussed the generation mechanism of bitter taste, the new methods of masking bitter taste and the evaluation me-thods of bitter taste, in order to provide references for the taste masking of Chinese patent medicines preparations.
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Medicamentos sin Prescripción , Gusto , Astringentes , China , Humanos , Medicina Tradicional China , Medicamentos sin Prescripción/farmacologíaRESUMEN
BACKGROUND: The goal of this study was to review eligible randomized controlled trials to determine the efficacy of the sinus tarsi approach (STA) versus the extended lateral approach (ELA) for the treatment of displaced intra-articular calcaneal fractures (DIACF). METHODS: Using appropriate keywords, we identified relevant studies using PubMed, the Cochrane Library, Embase, CNKI, VANFUN, and VIP. Key pertinent sources in the literature were also reviewed, and all articles published through June 2020 were considered for inclusion. For each study, we assessed odds ratios (ORs), mean difference (MD), and 95% confidence interval (95% CI) to assess and synthesize the outcomes. RESULTS: We included 15 RCTs, with a total of 847 patients in the STA group and 959 in the ELA group. The results found that after STA and ELA, no significant difference in changes of Böhler's angle (WMD: 0.746, 95% CI: - 0.316-1.809), Gissane angle (WMD: - 0.710, 95% CI: - 2.157-0.737), calcaneal heights (WMD: 0.378, 95% CI: - 1.973-2.728), calcaneal widths (SMD: - 0.431, 95% CI: - 1.604- 0.742), calcaneal lengths (WMD: 0.691, 95% CI: - 0.749-2.131). Besides, there was no significant difference in the incidence of complications between the STA group and the ELA group (RR: 0.592, 95% CI: 0.336-1.045). CONCLUSION: There was no difference in clinical efficacy between STA and ELA in treating DIACF. Besides, there is still a need of large-sample, high-quality, long-term randomized controlled trials to confirm the conclusion. LEVEL OF EVIDENCE: Level I-High-Quality Prospective Randomized Study.
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Calcáneo , Fracturas Óseas , Fracturas Intraarticulares , Calcáneo/cirugía , Fijación Interna de Fracturas , Talón , Humanos , Fracturas Intraarticulares/diagnóstico por imagen , Fracturas Intraarticulares/cirugía , Estudios Prospectivos , Ensayos Clínicos Controlados Aleatorios como Asunto , Resultado del TratamientoRESUMEN
Refrigeration is commonly used to extend the storage life of "Nanguo" pears, but fruit in long-term refrigeration is prone to peel browning, which is related to membrane lipid degradation. To determine the mechanism of membrane lipid degradation, we identified two R2R3-MYB transcription factors (TFs), PuMYB21 and PuMYB54, from "Nanguo" pears, which were notably expressed in response to cold stress and during the peel-browning process. The results from yeast one-hybrid, electrophoretic mobility shift, and transient expression assays indicated that both PuMYB21 and PuMYB54 directly bind to the promoter of PuPLDß1 (a key enzyme catalyzing the hydrolysis of membrane phospholipids) and activate its expression, which probably enhances the degradation of membrane phospholipids and eventually results in peel browning. Moreover, the overexpression of PuMYB21 and PuMYB54 can greatly activate the transcription of endogenous PuPLDß1 in both "Nanguo" pear fruits and calli, and their silencing can inhibit its transcription. Furthermore, yeast two-hybrid, bimolecular fluorescence complementation, and pull-down assays verified that PuMYB21 interacts with PuMYB54 to enhance the expression of PuPLDß1. In summary, we demonstrate that PuMYB21 and PuMYB54 may have roles in membrane lipid metabolism by directly binding to the downstream structural gene PuPLDß1 during the low temperature-induced peel browning of "Nanguo" pears.
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WHAT IS KNOWN AND OBJECTIVE: Thyroid follicular carcinoma is a malignant tumor from thyroid follicular epithelium, which is prone to involve capsular and vascular invasion. The present study was conducted in order to detect the expression of microRNA-155-5p (miR-155-5p) in thyroid follicular carcinoma with an attempt to analyze its involvement in apoptosis-related factors. METHODS: Forty-five patients with thyroid follicular carcinoma made up the observation group and 45 patients with thyroid follicular adenoma were included into the control group. Tissues of thyroid follicular carcinoma and thyroid follicular adenoma were obtained from the patients, and analysed for expression of miR-155-5p by real-time fluorescence quantitative PCR (qPCR). The expression of cysteine-containing aspartic acid protein hydrolase-3 (Caspase-3) in thyroid follicular carcinoma was detected with the use of Western Blot analyses. Immunohistochemical method was used to detect the expression of B-cell lymphoma protein-2 (Bcl-2) in thyroid follicular carcinoma. RESULTS: There was significant difference in the expression of miR-155-5p between the two groups (Observation vs Control: 1.46 ± 0.42 vs 0.98 ± 0.33 P < .05). The expression of miR-155-5p was significantly different in the maximum diameter of tumor, vascular invasion and neural invasion (maximum diameter of tumor <4 cm vs ≥4 cm: 1.36 ± 0.40 vs 1.68 ± 0.32, vascular invasion N vs Y: 1.35 ± 0.42 vs 1.69 ± 0.39, Neural invasion N vs Y: 1.35 ± 0.38 vs 1.70 ± 0.31 P < .05). However, there was no significant difference in the expression of miR-155-5p in terms of different gender, age and group with or without lymph node metastasis (P > .05). Based on survival analysis, patients with high expression of miR-155-5p experienced short survival time (median survival time was 45 months, P < .05). There was a negative correlation between miR-155-5p and Caspase-3 (r = -.50, P < .05). In addition, positive correlation was observed between miR-155-5p and Bcl-2 (r = .55, P < .05). WHAT IS NEW AND CONCLUSION: There was increased expression of miR-155-5p in thyroid follicular carcinoma. The abnormal expression of miR-155-5p may be an independent prognostic factor for thyroid follicular carcinoma associated with cell apoptosis.
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Adenocarcinoma Folicular/genética , Apoptosis/genética , MicroARNs/genética , Neoplasias de la Tiroides/genética , Adulto , Anciano , Anciano de 80 o más Años , Caspasa 3/genética , Proliferación Celular/genética , Femenino , Humanos , Metástasis Linfática/genética , Masculino , Persona de Mediana Edad , Glándula Tiroides/patologíaRESUMEN
OBJECTIVE: To investigate the clinicopathological and immunohistochemical features as well as diagnosis, differential diagnosis,treatment and prognosis of patients with primary renal non-hodgkin's lymphoma (PRNHL). METHODS: Clinical data of 2 patients with PRNHL from January 2013 to October 2017 were retrospectively analyzed. RESULTS: One of the two male patients PRNHL aged 51 years old, admitted to hospital for swelling and pain in the right waist, B-ultra sonic examination showed a very solid upper right renal space occupying lesion. CT showed the soft tissue mass of the right kidney about 10.5 cmx 8.6 cm. Preoperative diagnosis was a right kidney cancer subjected to radical surgery, and the postoperative pathological diagnosis was a right renal diffuse large B cell lymphoma (DLBCL). Expression of CD20, CD79a, Mum-1, BCL-2, BCL-6 in tumor cells was identified by inmmunohistochemistry, and Ki-67 proliferation index was 90%. Six courses of chemotherapy with R-CHOPE regimen were carried out supplemented local radiotherapy. The patients were followed up for 32 months, which were generally in good condition and stable. The another patient was 65 years old, no without obvious clinical symptoms, B-ultrasonic examination found a space occupying lesion in the left kidney. Abdomind CT scan plus enhancement showed the left kidneydisplayed uneven enhanced lesions of size 9 cm x 5 cm, the preoperative diagnosis was left kidney cancer, the pathological diagnosis was diagnosed as mall B cell lymphoma of the left kidney after the radical operation. Immunohistochemistry determined expression of CD20, CD79a, pax-5, BCL-2 in tumor cells and Ki-67 proliferation index 10%. 6 courses of chemotherapy with CHOPE regimen had been performed and followed up for 20 months, the conditions were generally good and stable. CONCLUSION: PRNHL is a rare extranodal non-Hodgkin lymphoma with atypical clinical symptoms characterized by lumbar swelling pain. It is easy to be misdiagnosed as renal cell carcinoma before operation. The diagnosis depends on pathology and immunohisto-chemistry. The prognosis of 2 patients was relatively good. PRNHL should be differetiated with renal sarcomatoid cancer, nephroblastoma and other diseases. Surgical resection plus chemotherapy is recommended.
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Linfoma no Hodgkin , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica , Ciclofosfamida , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
OBJECTIVE: To establish a novel method to isolate endothelial progenitor cellsï¼EPCï¼ from cryopreserved umbilical cord blood (cryoUCB), to investigate the biological characteristics of EPC and to improve the rate of EPC obtained from cryoUCB. METHODS: Twelve cryoUCB samples during 2000 to 2001 years were collected from allogeneic cord blood bank, cryoUCB was thawed rapidly in a water bath at 37 â, total nucleated cells (TNCs) were washed by phosphate-buffered saline (PBS). TNCs were seeded onto fibronectin-coated dishes to isolate EPC. Flow cytometry and immunofluorescence were used to identify EPC. The function of EPC was identified in vitro, such as the incorporation of Dil-Ac-LDL and FITC-UEA-I, the formation of capillary-like structure in matrigel, and the release of VEGF by ELISA. RESULTS: One to five cluster of cobble stone-like cells appeared at 2-3 weeks after seeding. Flow cytometric analysis showed that positive rates of CD31, CD34, CD144, and VEGFR (CD309) wereï¼92.91±5.20ï¼%, ï¼30.0±23.27ï¼%, ï¼88.55±3.83ï¼% and ï¼67.21±12.12ï¼% in passage 1 to passage 3 of EPC. EPC could uptake Dil-Ac-LDL and FITC-UEA-I, form capillary-like network on Matriget and release VEGF. CONCLUSION: EPC had been successfully isolated from cryopreserved umbilical cord blood by this method with high stability and reproducibility. EPC can be obtained in 85% frozen umbilical cord blood. This method may lay a foundation to supply abundant EPC for clinical application.
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Células Progenitoras Endoteliales , Sangre Fetal , Diferenciación Celular , Células Cultivadas , Reproducibilidad de los Resultados , Células MadreRESUMEN
Proteins, as the major executer for cell progresses and functions, its abundance and the level of post-translational modifications, are tightly monitored by regulators. Genetic perturbation could help us to understand the relationships between genes and protein functions. Herein, to explore the impact of the genome-wide interruption on certain protein, we developed a cell lysate microarray on kilo-conditions (CLICK) with 4837 knockout (YKO) and 322 temperature-sensitive (ts) mutant strains of yeast (Saccharomyces cerevisiae). Taking histone marks as examples, a general workflow was established for the global identification of upstream regulators. Through a single CLICK array test, we obtained a series of regulators for H3K4me3, which covers most of the known regulators in S. cerevisiae We also noted that several group of proteins are involved in negatively regulation of H3K4me3. Further, we discovered that Cab4p and Cab5p, two key enzymes of CoA biosynthesis, play central roles in histone acylation. Because of its general applicability, CLICK array could be easily adopted to rapid and global identification of upstream protein/enzyme(s) that regulate/modify the level of a protein or the posttranslational modification of a non-histone protein.
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Redes Reguladoras de Genes , Código de Histonas/genética , Saccharomyces cerevisiae/genética , Acilcoenzima A/metabolismo , Acilación , Química Clic , Histonas/metabolismo , Lisina/metabolismo , Metilación , Modelos Biológicos , Mutación/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Estrés FisiológicoRESUMEN
Icariside II (ICA II) is used in erectile dysfunction treatment. Adipose tissue-derived stem cells (ADSCs) are efficient at improving erectile function. This study aimed to explore the action mechanism of ADSCs in improving erectile function. ADSCs were isolated from the adipose tissues of rats. Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay. The expressions of mRNA and protein were determined separately through qRT-PCR and western blot. The endogenous expressions of related genes were regulated using recombinant plasmids and cell transfection. A Dual-Luciferase Reporter Assay was performed to determine the interaction between miR-34a and STAT3. Rat models with bilateral cavernous nerve injuries (BCNIs) were used to assess erectile function through the detection of mean arterial pressure (MAP) and intracavernosal pressure (ICP). ICA II promoted ADSCs' proliferation and differentiation to Schwann cells (SCs) through the inhibition of miR-34a. Suppressed miR-34a promoted the differentiation of ADSCs to SCs by upregulating STAT3. ICA II promoted the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway. The combination of ICA II and ADSCs preserved the erectile function of the BCNI model rats. ADSCs treated with ICA II markedly preserved the erectile function of the BCNI model rats, which was reversed through miR-34a overexpression. ICA II promotes the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway, contributing to erectile function preservation after the occurrence of a cavernous nerve injury.
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Tejido Adiposo/metabolismo , Medicamentos Herbarios Chinos/uso terapéutico , Disfunción Eréctil/etiología , Flavonoides/uso terapéutico , Hemangioma Cavernoso del Sistema Nervioso Central/complicaciones , Tejido Adiposo/citología , Adulto , Diferenciación Celular , Medicamentos Herbarios Chinos/farmacología , Flavonoides/farmacología , Humanos , Masculino , Células de Schwann , TransfecciónRESUMEN
Interleukin 17 (IL-17) plays an important role in the pathogenesis of cardiac interstitial fibrosis. In this study, we explored the role of interleukin-17 in the development of diabetic cardiomyopathy and the underlying mechanisms. The level of IL-17 increased in both the serum and cardiac tissue of diabetic mice. Knockout of IL-17 improved cardiac function of diabetic mice induced by streptozotocin (STZ), and significantly alleviated interstitial fibrosis as manifested by reduced collagen mRNA expression and collagen deposition evaluated by Masson's staining. High glucose treatment induced collagen production were abolished in cultured IL-17 knockout cardiac fibroblasts (CFs). The levels of long noncoding RNA-AK081284 were increased in the CFs treated with high glucose or IL-17. Knockout of IL-17 abrogated high glucose induced upregulation of AK081284. Overexpression of AK081284 in cultured CFs promoted the production of collagen and TGFß1. Both high glucose and IL-17 induced collagen and TGFß1 production were mitigated by the application of the siRNA for AK081284. In summary, deletion of IL-17 is able to mitigate myocardial fibrosis and improve cardiac function of diabetic mice. The IL-17/AK081284/TGFß1 signaling pathway mediates high glucose induced collagen production. This study indicates the therapeutic potential of IL-17 inhibition on diabetic cardiomyopathy disease associated with fibrosis.