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1.
Plant Dis ; 2021 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-34668407

RESUMO

American ginseng (Panax quinquefolium) is a valuable medicinal plant that is commercially cultivated in China. In May 2020, Sclerotinia root rot of American ginseng was observed on 4-year-old plants in Fusong County in northeastern China, which is the most important part of the country for American ginseng cultivation. The pathogen only infected the tuberous ginseng roots, with sclerotia tightly attached to the root surface. Infected roots, which were brownish and had a watery soft rotted appearance (Fig. 1), eventually became hollow and filled with sclerotia. There were no significant changes to the aboveground plant parts during the initial infection stage, but as the disease progressed, the foliage became discolored and wilted because of the damaged roots. More than 31% of the plants in a 30-ha field were infected. Symptomatic roots were collected and sclerotia were removed from the diseased tissue, immersed in 1% NaClO for 1 min, rinsed three times with sterile water, and placed on acidified potato dextrose agar (PDA) in Petri dishes. After an incubation in darkness at 20 °C for 2-3 days, 21 suspected Sclerotinia isolates were obtained. Isolates JH1 and JH2 were randomly selected for identification. On PDA, colonies produced sparse, white, and cottony aerial mycelia (i.e., wool-like appearance), with septate, branched, and hyaline hyphae. Within 4 days of incubation, the PDA surface was covered with white hyphae. Small and white sclerotial primordia formed 3 days later and were irregularly distributed in the middle and along the edge of the Petri dish. After maturing, the hardened and black sclerotia had an irregular shape and size, ranging from 1.4 × 1.5 to 4.1 × 7.5 mm (n = 50). Most of the sclerotia developed separately, with approximately 15-25 per plate (Fig. 2). On the basis of their morphology, the isolates were initially identified as Sclerotinia sp. (Mordue and Holliday 1976; Kohn 1979). Using the JH1 and JH2 rDNA internal transcribed spacer (ITS) region (GenBank accession no. MZ031405 and MZ031406) and the aspartyl protease gene specific to S. sclerotiorum (MZ292709 and MZ292710) in GenBank as queries, BLAST searches revealed that the sequences were respectively 99%-100% similar to S. sclerotiorum sequences KF859933 and AF271387. The primer pairs for amplifying the ITS region and the aspartyl protease gene were respectively ITS4/ITS5 (White et al. 1990) and SSaspr F/SSaspr R (Abd-Elmagid et al. 2013). The pathogenicity of JH1 and JH2 was evaluated using healthy plants. The roots of 4-year-old ginseng plants were washed, wiped with 75% alcohol, and transferred to flower pots containing sterile sand and sorghum grain (10:1 v/v) infested with 10-day-old isolates. For both isolates, 12 plants were inoculated, with four plants per pot. Control plants were transferred to flower pots containing sorghum grain lacking fungus. The inoculated samples were incubated in a greenhouse (12 h photoperiod and 25 °C) for 25 days before they were examined. The test was repeated twice. The inoculated roots exhibited the same symptoms as those observed in the field, whereas the controls remained symptomless. The same fungus was reisolated from all infected roots and resequencing results confirmed its identity. To the best of our knowledge, this is the first report of S. sclerotiorum causing Sclerotinia root rot on American ginseng in China. Because this disease is detrimental to the production of American ginseng, effective management strategies will need to be developed.

2.
Plant Dis ; 2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-34282929

RESUMO

Asian ginseng (Panax ginseng) is a valuable medicinal plant that is commercially cultivated in China. A long postharvest storage period is required before ginseng is processed. From October 2019 to May 2020, snow rot was observed on the roots of 4- and 5-year-old fresh ginseng stored in three cold storage facilities located in Tonghua and Changbai cities in northeastern China, which are the most important regions for Asian ginseng production. We sampled 1,000 ginseng roots from the three cold storage facilities, and the average disease incidence was 21%. Initially, sparse hyphae and microsclerotia appeared on the root epidermis. Lesions gradually softened and the epidermis detached easily. Multiple infected sites slowly converged, resulting in the formation of a dense complex of multiple sclerotia and thick hyphae on the surface of the ginseng root as well as internal decay. The infection eventually spread to the adjacent ginseng roots (Fig. 1). Sixteen diseased ginseng roots were collected and then sclerotia were removed from the root surface, immersed in 1% NaClO for 2 min, rinsed three times with sterile water, and placed on potato dextrose agar (PDA) containing streptomycin (40 µg/mL) in Petri dishes. After a 3-day incubation at 20 °C in darkness, 22 suspected Sclerotinia isolates were obtained. Isolates SN1 and SN2 were randomly selected for identification. On PDA, fast-growing colonies produced white, sparse, powdery, and cotton-like aerial mycelia, and the reverse side showed the same color (Fig. 2). Small and white sclerotial primordia formed 3 days later and a ring of sclerotia was detected at the plate periphery. At 7 to 10 days after incubation, the mature sclerotia were black, spherical-to-subspherical, and elongated or fused to form irregular shapes. Each Petri dish produced 55-65 sclerotia, measuring 1.1 × 1.2 to 3.2 × 3.9 mm (n = 100). The sclerotia were firmly attached to the agar surface. The isolates were initially identified as Sclerotinia sp. (Saito 1997). After sequencing the nuclear ribosomal internal transcribed spacer region (MW927134 and MW927135) and the ß-tubulin gene (MW929179 and MW929180) (White et al. 1990; Glass and Donaldson 1995), BLAST searches revealed 100% homology with JX262268 and JX296007 of the published S. nivalis strain KGC-S0601, respectively. The pathogenicity of the two isolates was tested using detached ginseng roots. Briefly, healthy roots were washed, surface-disinfested with 75% alcohol, and rinsed with sterile water. Mycelial plugs (5 mm diameter) removed from the margin of actively growing colonies on PDA were placed on the ginseng roots. For each isolate, four roots were inoculated, with two plugs per root. Additionally, PDA plugs without mycelia were used as the negative control. The roots were placed in a fresh-keeping box at 20 °C in darkness and evaluated after 7 days. The pathogenicity test was repeated twice. The symptoms on the inoculated roots were the same as those observed on the roots during cold storage, whereas the control roots remained symptomless. The same fungus was reisolated consistently from all infected roots and its identity was confirmed by resequencing, thereby fulfilling Koch's postulates. To the best of our knowledge, this is the first report of S. nivalis causing postharvest snow rot on Asian ginseng in China. The occurrence of this disease threatens the postharvest storage of Asian ginseng. Hence, effective management strategies must be developed.

3.
Front Endocrinol (Lausanne) ; 12: 614536, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33716970

RESUMO

Objective: Prediction of therapy response to intravenous methylprednisolone pulses (ivMP) is crucial for thyroid-associated ophthalmopathy (TAO). Image histograms may offer sensitive imaging biomarkers for therapy effect prediction. This study aimed to investigate whether pretherapeutic, multiparametric T2 relaxation time(T2RT) histogram features of extraocular muscles (EOMs) can be used to predict therapy response. Materials and Methods: Forty-five active and moderate-severe TAO patients, who were treated with standard ivMP and underwent orbital MRI before therapy, were retrospectively included in this study. The patients were divided into responsive (n = 24, 48 eyes) and unresponsive group(n = 21, 42 eyes) according to clinical evaluation. Baseline clinical features of patients and histogram-derived T2RT parameters of the EOMs were analyzed and compared. Logistic regression model was conducted to determine independent predictors, and a histogram features nomogram was formulated for personalized prediction. Results: Responsive group displayed lower values for 5th, 10th percentiles (P < 0.050, respectively), and higher values for 75th, 90th, and 95th percentiles, skewness, entropy, and inhomogeneity (P < 0.050, respectively) than unresponsive group. Multivariate logistic regression analysis showed that 95th percentile of >88.1 [odds ratio (OR) = 12.078; 95% confidence interval (CI) = 3.98-36.655, p < 0.001], skewness of >0.31 (OR = 3.935; 95% CI = 2.28-6.788, p < 0.001) and entropy of >3.41 (OR = 4.375; 95% CI = 2.604-7.351, p < 0.001) were independent predictors for favorable response. The nomogram integration of three independent predictors demonstrated optimal predictive efficiency, with a C-index of 0.792. Conclusions: Pre-treatment volumetric T2RT histogram features of EOMs could function to predict the response to ivMP in patients with TAO. The nomogram based on histogram features facilitates the selection of patients who will derive maximal benefit from ivMP.

4.
Zhongguo Gu Shang ; 33(11): 1048-52, 2020 Nov 25.
Artigo em Chinês | MEDLINE | ID: mdl-33269856

RESUMO

OBJECTIVE: To observe the clinical effect of platelet rich plasma (PRP) combined with ß tricalcium phosphate bioceramic bone in the treatment of non traumatic necrosis of the femoral head in ARCO stageⅡ. METHODS: From January 2017 to December 2018, 100 patients (160 hips) with ARCO stageⅡnon traumatic necrosis of the femoral head were divided into PRP group and control group. In PRP group, 50 patients (80 hips), 22 males and 28 females, aged from 18 to 65 (43.47± 7.23) years, with a course of 4 to 18 (15.8±2.9) months, underwent core decompression and bone grafting combined with PRP implantation. There were 50 cases (80 hips) in the control group, including 27 males and 23 females, aged 20 to 63 (45.72± 7.43) years, and the course of disease was 6 to 19 (14.9±3.8) months. Hip X-ay film was followed up after operation. Harris score and VAS score were used to evaluate the curative effect, and the survival rate of hip joint was recorded. RESULTS: All patients had good wound healing, no infection, thrombosis and other complications. All patients were followed up for 12 to 14 (12.0±0.4) months. Twelve months after operation, the image expression of PRP group was better than that of control group(P<0.05). Harris hip score and VAS score of pain at twelve months after operation were 89.98±6.17 and 1.68±1.02 in PRP group and 81.62±5.62 and 2.52±1.13 in control group, respectively. The survival rate of 96.25% in PRP group was significantly higher than 86.25% in control group. The postoperative score of two groups was higher than that before operation(P<0.05), but PRP group was better than control group at any time point statistical significance (P<0.05). CONCLUSION: Platelet-rich plasma(PRP) combined with artificialbone for core decompression and bone grafting can change the situation of simple artificial bone implantation and uncertain curative effect, improve the success rate of this operation, effectively reduce the collapse rate of femoral head necrosis in the early and middle stage, delay or even avoid hip replacement.


Assuntos
Necrose da Cabeça do Fêmur , Plasma Rico em Plaquetas , Adolescente , Adulto , Idoso , Artemisininas , Transplante Ósseo , Descompressão Cirúrgica , Feminino , Cabeça do Fêmur/cirurgia , Necrose da Cabeça do Fêmur/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Naftoquinonas , Resultado do Tratamento , Adulto Jovem
5.
Sci Rep ; 10(1): 19445, 2020 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-33173086

RESUMO

Using histogram analysis of T2 values to detect early involvement of extraocular muscles (EOMs) in patients with thyroid-associated ophthalmopathy (TAO). Five EOMs of each orbit were analyzed for 45 TAO patients and 22 healthy controls (HCs). Patients' EOMs were grouped into involved or normal-appearing EOMs (NAEOMs). Histogram parameters and signal intensity ratios (SIRs) of EOMs were compared; receiver operating characteristic (ROC) curve analysis was performed to differentiate NAEOMs from EOMs of HCs. 24 patients were reassessed following immunosuppressive treatment. For SIRs, involved muscles showed higher values than those of NAEOMs and HCs (p < 0.05); there were no differences between NAEOMs and HCs (p = 0.26). Parameters of involved muscles showed no different from those of NAEOMs excluding 25th, 50th percentiles, and standard deviation (SD) (p < 0.05). NAEOMs displayed higher values of 90th, 95th percentiles, SD, skewness, inhomogeneity, and entropy than HCs (p < 0.05). ROC curve analysis of entropy yielded the best area under the ROC curve (AUC; 0.816) for differentiating NAEOMs and HCs. After treatment, histogram parameters including 5th, 75th, 90th, and 95th percentiles, SD, kurtosis, inhomogeneity, and entropy were reduced in NAEOMs (p < 0.05). T2 histogram analysis could detect early involvement of EOMs in TAO prior to detection on conventional orbital MRI.


Assuntos
Imagem de Difusão por Ressonância Magnética/métodos , Oftalmopatia de Graves/diagnóstico , Interpretação de Imagem Assistida por Computador/métodos , Músculos Oculomotores/diagnóstico por imagem , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Estudos Retrospectivos
6.
Curr Med Sci ; 40(4): 608-613, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32767263

RESUMO

The corona virus disease 2019 (COVID-19) is an emerging respiratory infectious disease caused by SARS-CoV-2, which first occurred in December 2019 in Wuhan, China. These days, in China, chest CT is used for diagnosis of COVID-19, as an important complement to the reverse-transcription polymerase chain reaction (RT-PCR) test. Because of contacting with a large number of suspected or probable cases closely during chest CT examination, radiographers are easily infected with COVID-19. This article included the rearrangement of CT examination room in fever clinic, the rearrangement of human resources in radiology department, and the drafting of new operating procedures for radiologists who carry out CT examination on COVID-19 patients. This article also introduced the emergency management procedures of the department of radiology during the outbreak, and the experience of infection prevention for the staff of the department of radiology.


Assuntos
Betacoronavirus , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico por imagem , Pandemias , Pneumonia Viral/diagnóstico por imagem , Serviço Hospitalar de Radiologia/organização & administração , COVID-19 , Teste para COVID-19 , China/epidemiologia , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Desinfecção/normas , Humanos , Controle de Infecções/organização & administração , Controle de Infecções/normas , Profissionais Controladores de Infecções/organização & administração , Pandemias/prevenção & controle , Pneumonia Viral/epidemiologia , Pneumonia Viral/prevenção & controle , Radiologistas/organização & administração , SARS-CoV-2 , Tomografia Computadorizada por Raios X
7.
Mycorrhiza ; 30(4): 529-539, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32562087

RESUMO

To ensure long-term survival of epiphytic orchids through active reintroduction, more research on critical life cycle stages such as seed germination and seedling establishment are needed. In this study, we used in vitro germination experiments to investigate the role of mycorrhizal fungi in determining seed germination and growth in the endangered epiphytic orchid species, Dendrobium chrysotoxum. Symbiotic seed germination experiments were conducted for 90 days under different light conditions with fungal strains isolated from protocorms of D. chrysotoxum and three sister species. Molecular analyses showed that five strains belonged to the typical orchid mycorrhizal family Tulasnellaceae, whereas the other two strains belonged to the Sebacinaceae and the genus Coprinellus. Fungal inoculation, light conditions, and their interaction had a significant effect on protocorm formation and seedling development. Three fungal isolates, including two from D. chrysotoxum and one from D. catenatum, significantly stimulated protocorm formation and seedling development under light conditions. However, fungi isolated from host protocorms (GC-14 and GC-15) produced the highest number of seedlings after 50 days (49.5 ± 8.5%, 51.3 ± 9.0%, respectively), while the fungus isolated from D. catenatum protocorms produced the maximum number of seedlings only after 90 days (48.7 ± 16.1%). To conclude, this study has shown that light conditions and the identity of fungi had a strong effect on in vitro seed germination and seedling formation in an epiphytic orchid, with fungi isolated from host protocorms leading to accelerated germination and seedling formation. Therefore, fungal source should be taken into account when using seeds and compatible fungi for seedling propagation and in situ reintroduction.


Assuntos
Dendrobium , Micorrizas , Orchidaceae , Animais , China , Espécies em Perigo de Extinção , Germinação , Sementes , Simbiose
8.
Zhongguo Dang Dai Er Ke Za Zhi ; 22(1): 53-57, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31948525

RESUMO

OBJECTIVE: To study the changes in the serum levels of Chemerin and Omentin-1 in children with Kawasaki disease (KD) in the acute stage after intravenous immunoglobulin (IVIG) treatment and related clinical significance. METHODS: A total of 60 children who were diagnosed with KD from January 2015 to April 2019 were enrolled as subjects. Forty healthy children and 40 children with acute infectious diseases were enrolled as the healthy control group and the infection control group respectively. According to the sensitivity to IVIG treatment, the children with KD were divided into an IVIG sensitive group with 51 children and a non-IVIG sensitive group with 9 children. According to the presence or absence of coronary artery lesion, the children with KD were divided into a CAL group with 13 children and a non-CAL group with 47 children. ELISA was used to measure the serum levels of Omentin-1 and Chemerin before and after the treatment. RESULTS: The children with KD had significantly higher serum levels of Chemerin and Omentin-1 than the healthy control and infection control groups before treatment (P<0.05). After 48 hours of treatment, the IVIG sensitive group had a significant reduction in the serum level of Chemerin (P<0.05), while there was no significant change in the serum level of Omentin-1 after treatment (P>0.05). Before treatment, the non-IVIG sensitive group had a significantly higher serum level of Chemerin than the IVIG sensitive group (P<0.05), and the CAL group had a significantly higher serum level of Chemerin than the non-CAL group, while there was no significant difference in the serum level of Omentin-1 between the IVIG sensitive and non-IVIG sensitive groups, as well as between the CAL and non-CAL groups (P>0.05). CONCLUSIONS: High serum levels of Chemerin and Omentin-1 may play an important role in the development and progression of KD. Chemerin may be involved in the development of CAL in children with KD. The serum level of Chemerin may be used as a new index for predicting the sensitivity to IVIG treatment.


Assuntos
Síndrome de Linfonodos Mucocutâneos , Adipocinas , Quimiocinas , Criança , Doença da Artéria Coronariana , Humanos , Imunoglobulinas Intravenosas
9.
Front Microbiol ; 11: 618942, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33391250

RESUMO

Panax ginseng rusty root rot caused by the Ilyonectria species complex is a devastating disease, and it is one of the main factors contributing to the difficulty in continual cropping. Rusty root rot occurs in all ginseng fields, but little is known about the taxonomy of the fungal pathogen complex, especially Ilyonectria and Ilyonectria-like species. Rusty root rot samples were collected from commercial ginseng cultivation areas of China, and the pathogens were isolated and purified as single spores. Based on the combination analysis of multiple loci (rDNA-ITS, TUB, HIS3, TEF, ACT, LSU, RPB1, RPB2, and SSU) and morphological characteristics, the pathogens causing ginseng rusty root rot were determined. Fungal isolates were obtained from infected roots in 56 locations within main cultivation areas in China. A total of 766 strains were identified as Ilyonectria, Ilyonectria-like and Rhexocercosporidium species, including I. robusta (55.0%), I. communis (21.7%), I. mors-panacis (10.9%), I. pseudodestructans (2.0%), I. changbaiensis (1.3%), I. qitaiheensis (1.3%), Neonectria obtusispora (2.0%), Dactylonectria torresensis (0.5%), D. sp. (0.5%), and R. panacis (1.5%), and four novel species, Thelonectria ginsengicola (1.0%), T. jixiensis (1.0%), T. mulanensis (0.8%) and T. fusongensis (0.5%), with a total of 14 species. As the pathogen present in the highest proportion, I. robusta was the most prevalent and damaging species, unlike the pathogens reported previously. All of the examined strains were proven to cause ginseng rusty root rot. Our results indicate that the taxonomy of the fungal complex associated with ginseng rusty root rot includes Ilyonectria, Ilyonectria-like genera (Dactylonectria, Neonectria, and Thelonectria) and Rhexocercosporidium.

10.
Poult Sci ; 98(11): 5307-5314, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31222288

RESUMO

Infectious bursal disease (IBD) is one of the most prevalent infectious diseases caused by IBD virus (IBDV), which results in bursal necrosis and immunosuppression that cause severe damage to the immune system in chickens. Cytokines are important mediators and regulators of both types of host responses. In the present study, layer chickens were artificially challenged with IBDV, and the differential expression of inflammatory genes was explored by using quantitative real-time PCR, which offered basic data for further study of IBDV pathogenesis. Data showed that after IBDV infection, the virus load in the bursa of Fabricius (BF) peaked at 96 h and then gradually decreased. Compared with those of the negative-infected group, the mRNA expression levels of pro-inflammatory cytokines (interleukin [IL]-1ß, IL-6, IL-7, IL-8, tumor necrosis factor [TNF]-α, transforming growth factor [TGF]-ß) and anti-inflammatory cytokine IL-10 in the infected group increased to varying degrees at 12 to 192 h, respectively. Furthermore, the IL-1ß mRNA expression peaked at 48 h; the mRNA transcript levels of IL-6, IL-8, and IL-10 were the highest at 96 h; TNF-α mRNA expression peaked at 120 h; the IL-7 mRNA expression peaked at 144 h; and the TGF-ß mRNA transcript level was the highest at 192 h. Taken together, these observations indicated that along with the change pattern of IBDV proliferation in BF, the mRNA expression of cytokines (IL-1ß, IL-6, IL-7, IL-8, IL-10, TNF-α, TGF-ß) obviously increased, and the kinetics of each of these cytokines was different. The kinetics of IL-6/IL-10 mRNA expression ratio was significantly positively correlated with that of the virus load. These results suggest that IBDV infection seriously interferes with the natural immune response mediated by inflammatory cytokines in chickens.


Assuntos
Infecções por Birnaviridae/veterinária , Galinhas , Citocinas/genética , Citocinas/imunologia , Inflamação/veterinária , Doenças das Aves Domésticas/imunologia , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/imunologia , Infecções por Birnaviridae/genética , Infecções por Birnaviridae/imunologia , Infecções por Birnaviridae/virologia , Vírus da Doença Infecciosa da Bursa/fisiologia , Inflamação/genética , Inflamação/imunologia , Inflamação/virologia , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/virologia , Distribuição Aleatória
11.
Int J Mol Med ; 43(1): 371-381, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30387808

RESUMO

Post­translational modification via small ubiquitin­like modifier (SUMO) is involved in the regulation of various important cellular processes. SUMO modification can be regulated at the level of conjugation, and can also be reversed by the SUMO­specific proteases (SENPs). However, current studies of the regulation and function of SENP in lung development remain limited. In this study, the expression levels of SENP1 and SUMO1 were assessed during lung development in rats. SUMO1 modification occurred during lung development and changes in SENP1 expression were consistent with the changes in the presence of free SUMO1. In order to investigate the function of SENP1, alveolar type (AT) 2 cells were transfected with SENP1­targeting small interfering RNA, and the proliferation, apoptosis and differentiation function of AT2 cells was subsequently evaluated. Marked upregulation of conjugated SUMO1 was observed following SENP1 inhibition. Furthermore, depletion of SENP1 resulted in increased apoptosis, decreased proliferation and impaired differentiation status of AT2 cells. Thus, the results support that SENP1 is an essential regulator of the balance between SUMOylation and deSUMOylation during lung development, specifically affecting the proliferation and differentiation status of AT2 cells.


Assuntos
Células Epiteliais Alveolares/citologia , Células Epiteliais Alveolares/metabolismo , Diferenciação Celular , Cisteína Endopeptidases/fisiologia , Organogênese , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cisteína Endopeptidases/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Organogênese/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Proteína SUMO-1/genética , Proteína SUMO-1/metabolismo , Tretinoína/farmacologia , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética
12.
Zhongguo Dang Dai Er Ke Za Zhi ; 20(5): 403-409, 2018 May.
Artigo em Chinês | MEDLINE | ID: mdl-29764579

RESUMO

OBJECTIVE: To study the expression of SUMO-modified CCAAT enhancer binding protein α (C/EBPα) in preterm rat model of bronchopulmonary dysplasisa (BPD) induced by hyperoxia exposure and its role. METHODS: Eighteen preterm rats were randomly divided into an air group and a hyperoxia group (n=9 each). The model of BPD was prepared in preterm rats exposed to hyperoxia. The rats from the two groups were sacrificed on postnatal days 4, 7 and 14 respectively (3 rats at each time) and lung tissues were harvested. Periodic acid-Schiff (PAS) staining was used to observe the differentiation of rat lung tissues. Ki67 expression was detected by immunohistochemistry. Western blot was used to measure the protein expression of small ubiquitin-related modifier-1(SUMO1) and C/EBPα. A co-immunoprecipitation assay was performed to measure the protein expression of SUMO-modified C/EBPα. RESULTS: Compared with the air group, the hyperoxia group showed a decreased glycogen content in the lung tissue on postnatal day 4, and an increased content on postnatal days 7 and 14. Over the time of hyperoxia exposure, the hyperoxia group showed an increased expression of Ki67 in the lung tissue compared with the air group at all time points. Compared with the air group, the protein expression of C/EBPα increased on postnatal day 4 and decreased on postnatal days 7 and 14 in the hyperoxia group (P<0.05). The hyperoxia group had significantly upregulated expression of SUMO1 and SUMO-modified C/EBPα compared with the air group at all time points (P<0.05). In the hyperoxia group, the protein expression of SUMO-modified C/EBPα was positively correlated with the glycogen content (r=0.529, P<0.05) and the expression of Ki67 (r=0.671, P<0.05). CONCLUSIONS: Hyperoxia may induce over-proliferation and differentiation disorders of alveolar epithelial cells in preterm rat model of BPD, possibly through an increased expression of SUMO-modified C/EBP&alpha.


Assuntos
Displasia Broncopulmonar/etiologia , Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Hiperóxia/patologia , Sumoilação , Animais , Animais Recém-Nascidos , Displasia Broncopulmonar/metabolismo , Displasia Broncopulmonar/patologia , Proliferação de Células , Modelos Animais de Doenças , Hiperóxia/complicações , Antígeno Ki-67/análise , Alvéolos Pulmonares/patologia , Ratos , Ratos Sprague-Dawley
13.
Mitochondrial DNA B Resour ; 3(2): 1133-1134, 2018 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-33490563

RESUMO

In this study, the complete mitochondrial genome of three-spined stickleback, Gasterosteus aculeatus, was determined through sequencing of PCR fragments. The complete mitochondrial genome of G. aculeatus was 16,543 bp in length and encoded 13 protein-coding genes, 22 transfer RNA (tRNA) genes, and two ribosomal RNA genes. The overall nucleotide composition is: 27.0% A, 28.4% T, 27.4% C, and 17.2% G, with a total G + C content of 44.6%. By phylogenetic analysis using ML method, G. aculeatus showed the closest relationship with the blackspotted stickleback (Gasterosteus wheatlandi).

14.
Int J Mol Med ; 40(4): 1037-1046, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28902364

RESUMO

CCAAT enhancer binding protein alpha (C/EBPα) is a transcription factor regulating the core aspects of cell growth and differentiation. The present study investigated the level and functional role of C/EBPα during the development of the rat lung. C/EBPα protein exhibits a dynamic expression pattern. The correlation between the expression of C/EBPα protein and the content of glycogen during lung maturation was analyzed to understand the function of C/EBPα in lung differentiation. The high expression of C/EBPα coincides with the reduction of glycogen in the fetal lung. In addition, the authors identified that changes in the level of C/EBPα are associated with the secretion of pulmonary surfactant. C/EBPα is modified by small ubiquitin-related modifier (SUMO) post-translationally. The results of double immunofluorescence staining and immunoprecipitation demonstrated that SUMO-modified C/EBPα was present in the lung. The sumoylated C/EBPα gradually decreased during lung differentiation and was negatively correlated with pulmonary surfactant secretion, thereby suggesting that the SUMO modification may participate in C/EBPα-mediated lung growth and differentiation. These results indicated that C/EBPα played a role in lung development and provided the insight into the mechanism underlying SUMO-modification.


Assuntos
Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Pulmão/embriologia , Pulmão/metabolismo , Proteínas Associadas a Surfactantes Pulmonares/metabolismo , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Animais , Diferenciação Celular , Feminino , Glicogênio/metabolismo , Pulmão/citologia , Morfogênese , Fosfatidilcolinas/metabolismo , Fosfolipídeos/metabolismo , Ratos Sprague-Dawley
15.
Mol Med Rep ; 16(2): 1493-1501, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28586043

RESUMO

Hyperoxia is one of the primary causes of bronchopulmonary dysplasia, which may occur in premature infants following supplemental oxygen therapy. Glucose regulated protein 78 (GRP78), which is a molecular chaperone located in the lumen of the endoplasmic reticulum (ER), has been reported to regulate hyperoxia­associated ER stress. The role of GRP78 in lung epithelial cells during hyperoxia remains to be elucidated. In the present study, the A549 cultured human lung epithelial cell line was exposed to hyperoxic conditions, and then transfected with short interfering (si)RNA targeted to GRP78. siRNA or pEGFP­N1 plasmid were used to knockdown or overexpress specific genes, reverse transcription-quantitative polymerase chain reaction and western blot analysis were used to detect RNA and protein levels of gene expression, and flow cytometry was used to detect apoptosis. The expression levels of ER stress­associated genes were determined, and a significant increase in C/EBP homologous protein (CHOP) expression and apoptosis of A549 cells was observed, following GRP78 knockdown. The overexpression of CHOP downregulated B­cell lymphoma (Bcl)­2 expression levels, upregulated BCL2 associated X (Bax), and increased apoptosis of A549 cells under conditions of hyperoxia. CHOP knockdown demonstrated the opposite effect on Bcl­2 and Bax expression levels. These results suggested that GRP78 silencing promoted lung epithelial cell apoptosis during hyperoxia, via regulation of the CHOP pathway.


Assuntos
Células Epiteliais Alveolares/patologia , Apoptose , Inativação Gênica , Proteínas de Choque Térmico/metabolismo , Hiperóxia/metabolismo , Hiperóxia/patologia , Transdução de Sinais , Fator de Transcrição CHOP/metabolismo , Células A549 , Fator 6 Ativador da Transcrição/metabolismo , Células Epiteliais Alveolares/metabolismo , Endorribonucleases/metabolismo , Humanos , RNA Interferente Pequeno/metabolismo , Proteína X Associada a bcl-2/metabolismo , eIF-2 Quinase/metabolismo
16.
World J Pediatr ; 13(3): 228-235, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27995540

RESUMO

BACKGROUND: High-mobility group box-1 (HMGB1) protein acts as an important pro-infl ammatory mediator, which is capable of activating inflammation and tissue repair. HMGB1 can bind to its receptor such as advanced glycation end products (RAGE). RAGE, in turn, can promote the production of pro-inflammatory cytokines. Soluble RAGE (sRAGE) is a truncated form of the receptor comprising the extracellular domain of RAGE and can inhibit RAGE-activation. The objective of this study was to investigate whether HMGB1 and RAGE are involved in the development of brain injury in preterm infants. METHODS: In total, 108 infants ≤34 weeks gestation at birth were divided into 3 groups according to cranial altrasound scan: mild brain damage (n=33), severe brain damage (n=8) and no brain damage (n=67). All the placentas were submitted for pathologic evaluation. Histological chorioamnionitis (HCA) was defined as neutrophil infi ltration of amniotic membranes, umbilical cord or chorionic plate. Expressions of HMGB1 and RAGE proteins were assessed by immunohistochemical analysis. The concentration of HMGB1 and sRAGE in umbilical cord blood were measured by enzyme-linked immunosorbent assay. RESULTS: The frequency of HCA was 30.12%. HCA was associated with elevated concentrations of HMGB1 and decreased sRAGE in umbilical cord blood. The severe brain injury group demonstrated higher cord blood HMGB1 concentrations (P<0.001) and lower sRAGE concentrations (P<0.001) than both other groups. Brain injury in the premature infants was linked to intense staining for HMGB1/RAGE, particularly in infl ammatory cells. CONCLUSIONS: Changes of cord blood HMGB1 and sRAGE of premature infants had direct relationship with the degree of infl ammation and severity of brain damage. Monitoring sRAGE and HMGB1 levels may be helpful to predict intrauterine infection and brain injury in premature infants.


Assuntos
Lesões Encefálicas/congênito , Lesões Encefálicas/metabolismo , Produtos Finais de Glicação Avançada/metabolismo , Proteína HMGB1/metabolismo , Recém-Nascido Prematuro , Biomarcadores/metabolismo , Lesões Encefálicas/diagnóstico por imagem , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imuno-Histoquímica , Recém-Nascido , Masculino , Ultrassonografia
17.
J Huazhong Univ Sci Technolog Med Sci ; 36(5): 736-740, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27752909

RESUMO

The purpose of the study was to observe changes in endolymphatic hydrops by using intratympanic injection of gadolinium and magnetic resonance imaging (MRI) before and after endolymphatic sac surgery in patients with unilateral Meniere's disease. Thirteen patients with unilateral Meniere's disease undergoing endolymphatic sac surgery were retrospectively and prospectively analyzed. Three-dimensional fluid-attenuated inversion recovery or three-dimensional real inversion recovery MRI was performed 24 h after an intratympanic injection of gadolinium to grade the presence of endolymphatic hydrops. Among the 13 patients with hydrops confirmed by preoperative MRI, vestibular hydrops had no significant change in all patients; cochlear hydrops became negative in 2 patients, and remained unchanged in the other 11 patients after surgery. Definite vertigo attacks were substantially controlled in one patient and completely controlled in 12 patients during a follow-up period of 8-34 months after surgery. The hearing levels were improved in 3 patients, remained unchanged in 7 patients, and decreased in 3 patients. In conclusion, endolymphatic sac surgery does not always alleviate endolymphatic hydrops in patients with Meniere's disease. Relief from vertigo cannot always be attributed to the remission of hydrops. A change in hearing levels cannot be explained by hydrops status alone.


Assuntos
Hidropisia Endolinfática/diagnóstico por imagem , Saco Endolinfático/diagnóstico por imagem , Imageamento por Ressonância Magnética , Doença de Meniere/diagnóstico por imagem , Adulto , Idoso , Meios de Contraste/administração & dosagem , Hidropisia Endolinfática/patologia , Hidropisia Endolinfática/cirurgia , Saco Endolinfático/patologia , Saco Endolinfático/cirurgia , Feminino , Gadolínio/administração & dosagem , Humanos , Imageamento Tridimensional , Masculino , Doença de Meniere/patologia , Doença de Meniere/cirurgia , Pessoa de Meia-Idade
18.
Zhongguo Dang Dai Er Ke Za Zhi ; 18(9): 867-873, 2016 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-27655546

RESUMO

OBJECTIVE: To study the association between endoplasmic reticulum stress (ERS) pathway mediated by inositol-requiring kinase 1 (IRE1) and the apoptosis of type II alveolar epithelial cells (AECIIs) exposed to hyperoxia. METHODS: The primarily cultured AECIIs from preterm rats were devided into an air group and a hyperoxia group. The model of hyperoxia-induced cell injury was established. The cells were harvested at 24, 48, and 72 hours after hyperoxia exposure. An inverted phase-contrast microscope was used to observe morphological changes of the cells. Annexin V/PI double staining flow cytometry was performed to measure cell apoptosis. RT-PCR and Western blot were used to measure the mRNA and protein expression of glucose-regulated protein 78 (GRP78), IRE1, X-box binding protein-1 (XBP-1), and C/EBP homologous protein (CHOP). An immunofluorescence assay was performed to measure the expression of CHOP. RESULTS: Over the time of hyperoxia exposure, the hyperoxia group showed irregular spreading and vacuolization of AECIIs. Compared with the air group, the hyperoxia group showed a significantly increased apoptosis rate of AECIIs and significantly increased mRNA and protein expression of GRP78, IRE1, XBP1, and CHOP compared at all time points (P<0.05). The hyperoxia group had significantly greater fluorescence intensity of CHOP than the air group at all time points. In the hyperoxia group, the protein expression of CHOP was positively correlated with the apoptosis rate of AECIIs and the protein expression of IRE1 and XBP1 (r=0.97, 0.85, and 0.88 respectively; P<0.05). CONCLUSIONS: Hyperoxia induces apoptosis of AECIIs possibly through activating the IRE1-XBP1-CHOP pathway.


Assuntos
Apoptose , Estresse do Retículo Endoplasmático/fisiologia , Endorribonucleases/fisiologia , Hiperóxia/patologia , Complexos Multienzimáticos/fisiologia , Alvéolos Pulmonares/patologia , Animais , Células Cultivadas , Células Epiteliais/fisiologia , Feminino , Hiperóxia/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Transcrição CHOP/fisiologia , Proteína 1 de Ligação a X-Box/fisiologia
19.
Sci Rep ; 6: 27356, 2016 06 06.
Artigo em Inglês | MEDLINE | ID: mdl-27264613

RESUMO

Purple carrots (Daucus carota ssp. sativus var. atrorubens Alef.) accumulate large amounts of cyanidin-based anthocyanins in their taproots. Cyanidin can be glycosylated with galactose, xylose, and glucose in sequence by glycosyltransferases resulting in cyanidin 3-xylosyl (glucosyl) galactosides in purple carrots. The first step in the glycosylation of cyanidin is catalysis by UDP-galactose: cyanidin galactosyltransferase (UCGalT) transferring the galactosyl moiety from UDP-galactose to cyanidin. In the present study, a gene from 'Deep purple' carrot, DcUCGalT1, was cloned and heterologously expressed in E. coli BL21 (DE3). The recombinant DcUCGalT1 galactosylated cyanidin to produce cyanidin-3-O-galactoside and showed optimal activity for cyanidin at 30 °C and pH 8.6. It showed lower galactosylation activity for peonidin, pelargonidin, kaempferol and quercetin. It accepted only UDP-galactose as a glycosyl donor when cyanidin was used as an aglycone. The expression level of DcUCGalT1 was positively correlated with anthocyanin biosynthesis in carrots. The enzyme extractions from 'Deep purple' exhibited galactosylation activity for cyanidin, peonidin and pelargonidin, while those from 'Kuroda' (a non-purple cultivar) did not.


Assuntos
Antocianinas/metabolismo , Daucus carota/enzimologia , Galactose/metabolismo , Galactosiltransferases/metabolismo , Clonagem Molecular , Estabilidade Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Galactosiltransferases/química , Galactosiltransferases/genética , Expressão Gênica , Concentração de Íons de Hidrogênio , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidade por Substrato , Temperatura
20.
Pediatr Neurol ; 61: 94-98.e1, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27353694

RESUMO

BACKGROUND: To determine the association of histologic chorioamnionitis (HCA) and fetal inflammatory response syndrome (FIRS) with brain injuries in infants born to mothers with preterm premature rupture of membranes. METHODS: A total of 103 singleton infants born to mothers with preterm premature rupture of membranes were enrolled. The placental inflammation was confirmed by HCA, and FIRS was defined in fetuses with preterm labor and an elevation of the fetal plasma interleukin-6 concentration. Examination of brain images was conducted to confirm the existence of brain injuries. Based on placental HCA and umbilical cord blood interleukin-6 level, all patients were divided into three groups: HCA(-)FIRS(+), HCA(+)FIRS(-), and HCA(+)FIRS(+). RESULTS: Among all infants with preterm premature rupture of membranes, 53.40% were exposed to HCA, 20.38% experienced FIRS, and the overall incidence of brain injuries was 38.83%. The incidence of brain injury in HCA(-)FIRS(+), HCA(+)FIRS(-), and HCA(+)FIRS(+) groups were 20.83%, 41.18%, and 76.19%, respectively. HCA at the advanced grades and stages was associated with increased risk of brain injury. Umbilical cord blood levels of interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α), and granulocyte-colony stimulating factor (G-CSF) in premature infants with brain injuries were significantly higher than in those without brain injuries. Infants diagnosed with both HCA and FIRS showed significantly higher levels of IL-8, TNF-α, and G-CSF than those with HCA alone. CONCLUSIONS: Preterm infants exposed to severe chorioamnionitis had an increased risk of brain injury. IL-6, IL-8, TNF-α, and G-CSF in cord blood were associated with brain injuries in preterm infants and may be used as extradiagnostic criteria.


Assuntos
Lesões Encefálicas/sangue , Lesões Encefálicas/epidemiologia , Corioamnionite/sangue , Corioamnionite/epidemiologia , Recém-Nascido Prematuro/sangue , Adulto , Biomarcadores/sangue , Lesões Encefálicas/imunologia , Corioamnionite/imunologia , Corioamnionite/patologia , Feminino , Sangue Fetal/imunologia , Ruptura Prematura de Membranas Fetais , Humanos , Recém-Nascido de Baixo Peso/sangue , Recém-Nascido de Baixo Peso/imunologia , Recém-Nascido , Recém-Nascido Prematuro/imunologia , Masculino , Placenta/imunologia , Placenta/patologia , Gravidez , Estudos Prospectivos , Risco , Fatores de Risco
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