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1.
Planta ; 254(3): 50, 2021 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-34386845

RESUMO

MAIN CONCLUSION: Overexpression of the leaf color (Lc) gene in Ma bamboo substantially increased the accumulation level of anthocyanin, and improved plant tolerance to cold and drought stresses, probably due to the increased antioxidant capacity. Most bamboos, including Ma bamboo (Dendrocalamus latiflorus Munro), are naturally evergreen and sensitive to cold and drought stresses, while it's nearly impossible to make improvements through conventual breeding due to their long and irregular flowering habit. Moreover, few studies have reported bamboo germplasm innovation through genetic engineering as bamboo genetic transformation remains difficult. In this study, we have upregulated anthocyanin biosynthesis in Ma bamboo, to generate non-green Ma bamboo with increased abiotic stress tolerance. By overexpressing the maize Lc gene, a bHLH transcription activator involved in the anthocyanin biosynthesis in Ma bamboo, we generated purple bamboos with increased anthocyanin levels including cyanidin-3-O-rutinoside, peonidin 3-O-rutinoside, and an unknown cyanidin pentaglycoside derivative. The expression levels of 9 anthocyanin biosynthesis genes were up-regulated. Overexpression of the Lc gene improved the plant tolerance to cold and drought stress, probably due to increased antioxidant capacity. The levels of the cold- and drought-related phytohormone jasmonic acid in the transgenic plants were also enhanced, which may also contribute to the plant stress-tolerant phenotypes. High anthocyanin accumulation level did not affect plant growth. Transcriptomic analysis showed higher expressions of genes involved in the flavonoid pathway in Lc transgenic bamboos compared with those in wild-type ones. The anthocyanin-rich bamboos generated here provide an example of ornamental and multiple agronomic trait improvements by genetic engineering in this important grass species.


Assuntos
Secas , Regulação da Expressão Gênica de Plantas , Antocianinas , Resposta ao Choque Frio , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo
2.
World J Pediatr ; 17(4): 409-418, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34059960

RESUMO

BACKGROUND: Primary vesicoureteral reflux (VUR) is a common congenital anomaly of the kidney and urinary tract (CAKUT) in childhood. The present study identified the possible genetic contributions to primary VUR in children. METHODS: Patients with primary VUR were enrolled and analysed based on a national multi-center registration network (Chinese Children Genetic Kidney Disease Database, CCGKDD) that covered 23 different provinces/regions in China from 2014 to 2019. Genetic causes were sought using whole-exome sequencing (WES) or targeted-exome sequencing. RESULTS: A total of 379 unrelated patients (male: female 219:160) with primary VUR were recruited. Sixty-four (16.9%) children had extrarenal manifestations, and 165 (43.5%) patients showed the coexistence of other CAKUT phenotypes. Eighty-eight patient (23.2%) exhibited impaired renal function at their last visit, and 18 of them (20.5%) developed ESRD at the median age of 7.0 (IQR 0.9-11.4) years. A monogenic cause was identified in 28 patients (7.39%). These genes included PAX2 (n = 4), TNXB (n = 3), GATA3 (n = 3), SLIT2 (n = 3), ROBO2 (n = 2), TBX18 (n = 2), and the other 11 genes (one gene for each patient). There was a significant difference in the rate of gene mutations between patients with or without extrarenal complications (14.1% vs. 6%, P = 0.035). The frequency of genetic abnormality was not statistically significant based on the coexistence of another CAKUT (9.6% vs. 5.6%, P = 0.139, Chi-square test) and the grade of reflux (9.4% vs. 6.7%, P = 0.429). Kaplan-Meier survival curve showed that the presence of genetic mutations did affect renal survival (Log-rank test, P = 0.01). PAX2 mutation carriers (HR 5.1, 95% CI 1.3-20.0; P = 0.02) and TNXB mutation carriers (HR 20.3, 95% CI 2.4-168.7; P = 0.01) were associated with increased risk of progression to ESRD. CONCLUSIONS: PAX2, TNXB, GATA3 and SLIT2 were the main underlying monogenic causes and accounted for up to 46.4% of monogenic VUR. Extrarenal complications and renal function were significantly related to the findings of genetic factors in children with primary VUR. Like other types of CAKUT, several genes may be responsible for isolated VUR.

3.
J Med Genet ; 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33323469

RESUMO

BACKGROUND: Nephronophthisis-related ciliopathies (NPHP-RC) account for the majority of cases of monogenetically caused end-stage renal disease (ESRD) in children. Exploring the correlation between the phenotype and genotype of NPHP-RC is helpful for early diagnosis and management. We investigated the phenotype and genotype spectra of NPHP-RC in a Chinese multicentre cohort. METHODS: Crosss-ectional and longitudinal data of 60 patients from 57 families with pathogenic NPHP-RC gene mutations distributed in 22 regions of China were collected into a unified, anonymous database. The mean observation time of this cohort was 3.5±3.1 years. RESULTS: Mutations in NPHP1 and NPHP3 were the most common genetic defects. Overall, 45% of patients presented with isolated nephronophthisis (NPH), and 55% exhibited the extrarenal phenotype, which frequently involved the liver (41.7%, n=25), central nervous system (26.7%, n=16), eyes (26.7%, n=16) and skeletal system (11.7%, n=7). Accidental detection of elevated serum creatinine and non-specific symptoms caused by chronic kidney disease occurred in 65% of patients. Patients carrying NPHP1 mutations mainly presented with isolated NPH (90%, 18/20) and progressed to ESRD at a mean age of 12.9±0.5 years. The mean age of ESRD onset in the non-NPHP1 group was lower than that in the NPHP1 group (6.2±1.4 years, p<0.001), especially for patients carrying NPHP3 mutations (3.1±1.2 years), showing a heterogeneous phenotype characterised by Bardet-Biedl syndrome (12.5%, n=5), Joubert syndrome (7.5%, n=3), COACH syndrome (2.5%, n=1), Mainzer-Saldino syndrome (2.5%, n=1), short-rib thoracic dysplasia (2.5%, n=1) and unclassified symptoms (32.5%, n=13). CONCLUSIONS: The Chinese Children Genetic Kidney Disease Database registry characterised the spectrum of the phenotype and genotype of NPHP-RC in the Chinese population. NPHP1 and NPHP3 were the most common pathogenic genes. Rapid progression to ESRD and liver involvement were noted in patients with NPHP3 mutations.

4.
Opt Express ; 28(23): 33809-33822, 2020 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-33182861

RESUMO

We proposed a differential fiber-optic SPR remote sensor with ultra-high sensitivity in telecom band. The working band of the sensor is designed as the C-band which is the low loss band of optical fiber communication aiming to improve the sensitivity and enable the capability of remote monitoring. The sensor head is a BK7 prism coated with Au/TiO2 films, enabling two channels for differential intensity interrogation. The intensities of the reflected lights through the channels vary oppositely within the measurement range of refractive index. Due to the sharp dip of angular resonant response in the C-band, the differential signal produces a steep slope as the refractive index of the sample varies, thus higher sensitivity is expected in a narrow measurement range. According to the results, the sensitivity is as high as 456 V/RIUs within the narrow measurement range of 1.3×10-2 RIUs and the resolution reaches to 6×10-6 RIUs. The measurement range can be tuned conveniently by adjusting the thickness of TiO2 film and can be expanded by increasing the number of sensing channels, which provides great convenience for the application of biosensor requiring high sensitivity.

5.
Eur J Med Genet ; 63(11): 104047, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32891756

RESUMO

BACKGROUND: WT1 mutations cause a wide spectrum of renal and extrarenal manifestations concerning urogenital development and the development of tumors. METHODS: We retrospectively collected the information on the genotype and phenotype of WT1 nephropathy from the multicenter registry since 2014 to 2019. All patients were stratified by renal function decline status or by sequence timing. Rapid progressive group was defined as rapidly developing into ERSD within 12 months since disease onset. Early sequencing group was defined as gene mutation identified before ERSD. RESULTS: Thirty-three (3.5%) cases were identified with a WT1 mutation in patients with steroid resistant nephrotic syndrome (SRNS), proteinuria and chronic kidney disease (CKD) 3-5 stage of unknown origin. ESRD developed in twenty patients at a median age of 4.3 years old. Comparing study between the rapid progressive group (n = 8) and non-rapid progressive group (n = 25) showed no significant difference in age of onset, gender, syndrome phenotype, genotype and proteinuria except for initial estimated glomerular filtration rate (eGFR) (p = 0.021) or sequencing timing (p = 0.003). In multivariable logistic regression analysis, the delayed sequencing was associated with rapid renal function decline, even after adjusting for established clinical factors including syndromic phenotype, genotype, age onset and eGFR at initial stage (p = 0.019). The renal survival analysis did not show a significantly better outcome in early sequencing group than in delayed sequencing group (p > 0.05). CONCLUSION: Screening for WT1 mutations should be performed in children with Wilms' tumor, proteinuria/SRNS or CKD. Early diagnosis of WT1 nephropathy through clinical and genetic findings is warranted.


Assuntos
Testes Genéticos/normas , Síndrome Nefrótica/diagnóstico , Proteinúria/diagnóstico , Insuficiência Renal Crônica/diagnóstico , Proteínas WT1/genética , Tumor de Wilms/diagnóstico , Pré-Escolar , Estudos de Coortes , Diagnóstico Precoce , Feminino , Testes Genéticos/métodos , Humanos , Lactente , Masculino , Estudos Multicêntricos como Assunto , Síndrome Nefrótica/genética , Proteinúria/genética , Insuficiência Renal Crônica/genética , Tumor de Wilms/genética
6.
Clin J Am Soc Nephrol ; 15(9): 1259-1266, 2020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32665227

RESUMO

BACKGROUND AND OBJECTIVES: During the coronavirus disease 2019 outbreak, the treatment of families with children on long-term KRT is challenging. This study was conducted to identify the current difficulties, worries regarding the next 2 months, and mental distress experienced by families with children on long-term KRT during the coronavirus disease 2019 outbreak and to deliver possible management approaches to ensure uninterrupted treatment for children on long-term KRT. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A multicenter online survey was conducted between February 10 and 15, 2020, among the families with children on long-term KRT from five major pediatric dialysis centers in mainland China. The primary caregivers of children currently on long-term KRT were eligible and included. Demographic information, severe acute respiratory syndrome coronavirus 2 infection status, current difficulties, and worries regarding the next 2 months were surveyed using a self-developed questionnaire. The Patient Health Questionnaire-9 and the General Anxiety Disorder Scale-7 were used to screen for depressive symptoms and anxiety, respectively. RESULTS: Among the children in the 220 families included in data analysis, 113 (51%) children were on dialysis, and the other 107 (49%) had kidney transplants. No families reported confirmed or suspected cases of coronavirus disease 2019. Overall, 135 (61%) and 173 (79%) caregivers reported having difficulties now and having worries regarding the next 2 months, respectively. Dialysis supply shortage (dialysis group) and hard to have blood tests (kidney transplantation group) were most commonly reported. A total of 29 (13%) caregivers had depressive symptoms, and 24 (11%) had anxiety. After the survey, we offered online and offline interventions to address their problems. At the time of the submission of this paper, no treatment interruption had been reported. CONCLUSIONS: The coronavirus disease 2019 outbreak has had physical, mental, logistical, and financial effects on families with children on long-term KRT.


Assuntos
Betacoronavirus/patogenicidade , Infecções por Coronavirus/prevenção & controle , Família/psicologia , Nefropatias/terapia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Terapia de Substituição Renal , Adaptação Psicológica , Adolescente , Adulto , Fatores Etários , COVID-19 , Cuidadores/psicologia , Criança , China/epidemiologia , Infecções por Coronavirus/psicologia , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Efeitos Psicossociais da Doença , Relações Familiares , Feminino , Pesquisas sobre Serviços de Saúde , Conhecimentos, Atitudes e Prática em Saúde , Acesso aos Serviços de Saúde , Interações entre Hospedeiro e Microrganismos , Humanos , Nefropatias/psicologia , Masculino , Saúde Mental , Pessoa de Meia-Idade , Segurança do Paciente , Pneumonia Viral/psicologia , Pneumonia Viral/transmissão , Pneumonia Viral/virologia , Terapia de Substituição Renal/efeitos adversos , Medição de Risco , Fatores de Risco , SARS-CoV-2 , Fatores de Tempo , Resultado do Tratamento
7.
Mol Genet Genomic Med ; 8(8): e1360, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32543055

RESUMO

BACKGROUND: Mutations in COQ8B (*615567) as a defect of coenzyme Q10 (CoQ10) cause steroid resistant nephrotic syndrome (SRNS). METHODS: To define the clinical course and prognosis of COQ8B nephropathy, we retrospectively assessed the genotype and phenotype in patients with COQ8B mutations from Chinese Children Genetic Kidney Disease Database. We performed the comparing study of renal outcome following CoQ10 treatment and renal transplantation between early genetic detection and delayed genetic detection group. RESULTS: We identified 20 (5.8%) patients with biallelic mutations of COQ8B screening for patients with SRNS, non-nephrotic proteinuria, or chronic kidney disease (CKD) of unknown origin. Patients with COQ8B mutations showed a largely renal-limited phenotype presenting with proteinuria and/or advanced CKD at the time of diagnosis. Renal biopsy uniformly showed focal segmental glomerulosclerosis. Proteinuria was decreased, whereas the renal function was preserved in five patients following CoQ10 administration combined with angiotensin-converting enzyme (ACE) inhibitor. The renal survival analysis disclosed a significantly better outcome in early genetic detection group than in delayed genetic detection group (Kaplan-Meier plot and log rank test, p = .037). Seven patients underwent deceased donor renal transplantation without recurrence of proteinuria or graft failure. Blood pressure showed decreased significantly during 6 to 12 months post transplantation. CONCLUSIONS: COQ8B mutations are one of the most common causes of adolescent-onset proteinuria and/or CKD of unknown etiology in the Chinese children. Early detection of COQ8B nephropathy following CoQ10 supplementation combined with ACE inhibitor could slow the progression of renal dysfunction. Renal transplantation in patients with COQ8B nephropathy showed no recurrence of proteinuria.


Assuntos
Testes Genéticos/métodos , Síndrome Nefrótica/congênito , Fenótipo , Proteínas Quinases/genética , Adolescente , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Criança , Diagnóstico Precoce , Feminino , Rejeição de Enxerto/epidemiologia , Humanos , Rim/metabolismo , Rim/patologia , Transplante de Rim/efeitos adversos , Transplante de Rim/métodos , Masculino , Mutação , Síndrome Nefrótica/diagnóstico , Síndrome Nefrótica/tratamento farmacológico , Síndrome Nefrótica/genética , Síndrome Nefrótica/terapia , Complicações Pós-Operatórias/epidemiologia , Ubiquinona/análogos & derivados , Ubiquinona/uso terapêutico
8.
Clin Genet ; 96(5): 402-410, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31328266

RESUMO

To explore the approaches and diagnostic yield of genetic testing for renal disease in children, we describe the genotype and phenotype of the national cohort of children with renal disease from 13 different regions of China recruited from 2014 to 2018 by building up the multicenter registration system (Chinese Children Genetic Kidney Disease Database, CCGKDD). Genetic diagnosis was confirmed in 42.1% of our cohort of 1001 pediatric patients with clinical suspicion of a genetic renal disease. Of the 106 distinct monogenetic disorders detected, 15 accounted for 60.7% of genetic diagnoses. The diagnostic yield was 29.1% in steroid resistant nephritic syndrome (SRNS), 61.4% in cystic renal disease, 17.0% in congenital anomalies of the kidney and urinary tract (CAKUT), 62.3% in renal tubular disease/renal calcinosis, and 23.9% for chronic kidney disease (CKD) 3 to 5 stage with unknown origin. Genetic approaches of target gene sequence (TGS), singleton whole-exome sequencing (WES) and trio-WES were performed with diagnostic rates of 44.8%, 36.2%, and 42.6%, respectively. The early use of trio-WES could improve the diagnostic rate especially in renal tubular disease and calcinosis. We report the genetic spectrum of Chinese children with renal disease. Establishment of the CCGKDD will improve the genetic work on renal disease.


Assuntos
Exoma/genética , Predisposição Genética para Doença , Doenças Renais Císticas/genética , Insuficiência Renal Crônica/genética , Criança , Pré-Escolar , China/epidemiologia , Estudos de Coortes , Feminino , Testes Genéticos , Humanos , Rim/metabolismo , Rim/patologia , Doenças Renais Císticas/diagnóstico , Doenças Renais Císticas/patologia , Masculino , Fenótipo , Insuficiência Renal Crônica/diagnóstico , Insuficiência Renal Crônica/patologia , Sistema Urinário/metabolismo , Sistema Urinário/patologia , Sequenciamento Completo do Exoma
10.
Clin Nephrol ; 92(2): 89-94, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31131822

RESUMO

OBJECTIVE: Nephronophthisis (NPH) is an autosomal recessive cystic kidney disease. Its onset is obscure, and its early clinical manifestations and pathological changes lack specificity, which makes clinical diagnosis difficult. At present, as many as 90 genetic alterations can result in NPH, which exhibits significant genetic heterogeneity. Therefore, high-throughput sequencing technology provides an effective method to identify and characterize novel NPH pathogenic genes when compared to Sanger sequencing. This study summarizes the gene mutations and clinical data of whole exome sequencing, which was used to diagnose 5 NPH patients to improve the understanding of the causative genes and clinical phenotypes of NPH. MATERIALS AND METHODS: The clinical manifestations, laboratory examination indexes, and imaging data of 5 patients of NPH were reported. Whole exome sequencing was performed in 5 children, and the causative genes and mutation sites were analyzed by bioinformatics and genetics. The mutation sites were verified in children and their parents using Sanger direct sequencing. RESULTS: Among the 5 patients (3 male and 2 female), 2 patients had infantile NPH, and 3 patients had juvenile NPH. The 2 infantile NPH patients were characterized by the onset of liver dysfunction accompanied by hypertension and left ventricular change, and the renal function progressed to end-stage renal disease (ESRD) after 7 months and 9 months, respectively. The 2 cases of infantile NPH had NPHP3 mutations, with one carrying compound heterozygous mutations (c.1358A>G, c.2369A>G) and the other simultaneously carrying a c.1174C>T IVS26-3A>G cleavage site mutation from the father and a nonsense mutation (p.392R>X, 939) from the mother. The 2 juvenile NPH children had entered ESRD at the onset of the disease, including 1 patient with Joubert syndrome. The 2 patients with juvenile NPH had frameshift mutations (c.1583 to 1596: deletion) and homozygous point mutations (7 c.640G>T) of the NPHP1 gene. In addition, another patient with frequent urination and nocturia resulting in stage CKD3 renal function had a complex heterozygous mutation of the NPHP2 gene (c.2686G>A, c.1943A>G). The urine A1MU/creatinine and urinary transferrin increased in all 5 patients without hematuria. CONCLUSION: Whole exome sequencing identified the causative genes of NPH in 5 children. In NPH children with NPHP3 gene mutations, renal functional damage was characterized by early onset and rapid progression to ESRD, often accompanied by liver dysfunction and hypertension.


Assuntos
Doenças Renais Císticas/congênito , Proteínas Adaptadoras de Transdução de Sinal/genética , Adolescente , Criança , Pré-Escolar , China , Proteínas do Citoesqueleto , Feminino , Heterozigoto , Homozigoto , Humanos , Lactente , Doenças Renais Císticas/diagnóstico , Doenças Renais Císticas/genética , Falência Renal Crônica/etiologia , Cinesina/genética , Masculino , Proteínas de Membrana/genética , Mutação/genética , Fenótipo , Fatores de Transcrição/genética , Sequenciamento Completo do Exoma
12.
Chem Commun (Camb) ; 54(81): 11411-11414, 2018 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-30246199

RESUMO

We report herein the identification of urea structure-like small molecules by structure-based virtual screening of 10.5 million compounds. Based on a variety of HEK-Blue hTLRs reporter cell assay results, we validated a TLR1/2-specific small molecule agonist, ZINC666243 (SMU127), with EC50 of 0.55 ± 0.01 µM. SMU127 stimulates NF-κB activation and promotes TNFα secretion in human macrophages and mononuclear cells. Moreover, the in vivo assay indicated that SMU127 could inhibit the growth of breast cancer tumors in BABL/c mice. This work has shown for the first time that a small molecule TLR1/2 agonist can inhibit breast cancer in vivo.

13.
Bioorg Med Chem ; 26(8): 2041-2050, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29534935

RESUMO

Toll-like receptor 2 (TLR2) can recognize pathogen-associated molecular patterns to defense against invading organisms and has been represents an attractive therapeutic target. Until today, none TLR2 small molecule antagonist have been developed in clinical trial. Herein, we designed and synthesized 50 N-benzylideneaniline compounds with the help of CADD. And subsequent in vitro studies leading to the optimized compound SMU-A0B13 with most potent inhibitory activity to TLR2 (IC50=18.21 ±â€¯0.87 µM). Preliminary mechanism studies indicated that this TLR2 inhibitor can work through the NF-κB signaling pathway with high specificity and low toxicity, and can also efficiently downregulate inflammatory cytokines, such as SEAP, TNF-α and NO in HEK-Blue hTLR2, human PBMC and Raw 264.7 cell lines. Additionally, the docking situation also indicate SMU-A0B13 can well bind to the TLR2-TIR (PDB: 1FYW) active domain, which probably explains the bioactivity.


Assuntos
Compostos de Anilina/química , Receptor 2 Toll-Like/antagonistas & inibidores , Compostos de Anilina/metabolismo , Compostos de Anilina/farmacologia , Animais , Sítios de Ligação , Domínio Catalítico , Células Cultivadas , Citocinas/metabolismo , Humanos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Camundongos , Simulação de Acoplamento Molecular , NF-kappa B/metabolismo , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacos , Relação Estrutura-Atividade , Receptor 2 Toll-Like/metabolismo
14.
J Oral Maxillofac Surg ; 76(1): 221-228, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28651068

RESUMO

PURPOSE: The aim of this study was to explore the effect of Choukroun platelet-rich fibrin (PRF) combined with autologous micro-morselized bone on the repair of mandibular defects in rabbits. MATERIALS AND METHODS: Thirty-six healthy New Zealand rabbits were selected for the present study. After models of mandibular defects were established, rabbits were randomly divided into Choukroun PRF, autologous micro-morselized bone (autologous), Choukroun PRF combined with autologous bone (combined) and model groups. After the rabbits were sacrificed at 2, 8, and 12 weeks postoperatively, their bone formation was assessed by x-ray and scanning electron microscopy, and the histologic changes of the mandibular defect area were detected by hematoxylin and eosin staining. Cone-beam computed tomography was used to observe the size of the change of the mandibular defect area. Bone mineral density (BMD) was analyzed by dual-energy x-ray absorptiometry. RESULTS: The bone defect in the combined group showed better repair, increased bone mineral content, and denser callus than the other groups, and the defect area was filled with mature trabecular bone. In the Choukroun PRF and autologous groups, the defect area was smaller and filled with osteoporotic trabecular bone. A clear mandibular defect area was still observed in the model group. Compared with the other groups, the combined group showed more bone regeneration, more fibrous tissue regeneration, and greater bone maturity at all time points. The combined group had the highest BMD, there was no relevant difference in BMD between the Choukroun PRF and autologous groups, and the model group had the lowest BMD. BMD in all 4 groups increased with time. CONCLUSION: These findings indicate that Choukroun PRF combined with autologous micro-morselized bone can substantially improve the repair of mandibular defects in rabbits, and the effect is superior to Choukroun PRF or autologous micro-morselized bone alone.


Assuntos
Transplante Ósseo/métodos , Mandíbula/cirurgia , Fibrina Rica em Plaquetas , Animais , Densidade Óssea , Regeneração Óssea , Tomografia Computadorizada de Feixe Cônico , Microscopia Eletrônica de Varredura , Coelhos , Transplante Autólogo
15.
Med Sci Monit ; 23: 4601-4611, 2017 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-28945699

RESUMO

BACKGROUND This study explored the effects of nano-hydroxyapatite/polyetheretherketone (n-HA/PEEK)- coated sandblasted, large-grit, and acid-etched (SLA) implants on inflammatory cytokines and osseointegration in peri-implantitis model beagle dogs. MATERIAL AND METHODS Peri-implantitis models were established. Eight beagle dogs were randomly and evenly assigned into SLA tied, SLA + n-HA/PEEK tied, SLA untied, or SLA + n-HA/PEEK untied groups. A special periodontal probe was used to detect the plaque index (PLI), probing depth (PD), and modified Sulcus Bleeding Index (mSBI). Gingival crevicular fluid was collected and an ELISA kit was utilized to detect IL-1, IL-6, and IL-17 levels. The colony-forming units were counted and the maximum shear strength of implants was tested using the axial pullout test. HE staining was used to detect the inflammation of peri-implant bone tissues. Osseointegration was observed through toluidine blue staining. Bone-to-implant contact (BIC) was obtained through histological observation and the mineral apposition rate (MAR) was calculated after immune fluorescent double staining. RESULTS The SLA tied group demonstrated higher levels of PLI, PD, mSBI, IL-1, IL-6, and IL-17 and a higher degree of inflammation than the SLA + n-HA/PEEK tied group. The tied groups also displayed similar results over the untied groups at the same time point. The maximum shear strength, BIC, and MAR in the SLA tied group were significantly lower than in the SLA + n-HA/PEEK tied group. CONCLUSIONS Our findings demonstrate that SLA + n-HA/PEEK implants can promote osseointegration and relieve the inflammation response of peri-implantitis in beagle dogs.


Assuntos
Condicionamento Ácido do Dente , Citocinas/metabolismo , Implantes Dentários , Durapatita/farmacologia , Cetonas/farmacologia , Nanopartículas/química , Osseointegração/efeitos dos fármacos , Peri-Implantite/metabolismo , Polietilenoglicóis/farmacologia , Animais , Osso e Ossos/patologia , Ensaio de Unidades Formadoras de Colônias , Placa Dentária/patologia , Modelos Animais de Doenças , Cães , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Minerais/metabolismo , Peri-Implantite/patologia , Resistência ao Cisalhamento
16.
Front Plant Sci ; 8: 1298, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28798758

RESUMO

Genetic engineering technology has been successfully used in many plant species, but is limited in woody plants, especially in bamboos. Ma bamboo (Dendrocalamus latiflorus Munro) is one of the most important bamboo species in Asia, and its genetic improvement was largely restricted by the lack of an efficient regeneration and transformation method. Here we reported a plantlet regeneration and Agrobacterium-mediated transformation protocol by using Ma bamboo young shoots as explants. Under our optimized conditions, embryogenic calluses were successfully induced from the excised young shoots on callus induction medium and rapidly grew on callus multiplication medium. Shoots and roots were regenerated on shoot induction medium and root induction medium, respectively, with high efficiency. An Agrobacterium-mediated genetic transformation protocol of Ma bamboo was established, verified by PCR and GUS staining. Furthermore, the maize Lc gene under the control of the ubiquitin promoter was successfully introduced into Ma bamboo genome and generated an anthocyanin over-accumulation phenotype. Our methods established here will facilitate the basic research as well as genetic breeding of this important bamboo species. Key achievements: A stable and high efficiency regeneration and Agrobacterium-mediated transformation protocol for Ma bamboo from vegetative organ is established.

17.
Cell Prolif ; 50(3)2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28205268

RESUMO

INTRODUCTION: This study aimed to investigate the functions of delta-like homologue 1 (DLK1) in the proliferation and differentiation of human dental pulp stem cells (hDPSCs). METHODS: Immunohistochemical analysis was used to determine the expression of alkaline phosphatase (ALP), dentin sialophosphoprotein (DSPP), DLK1, NOTCH1 and p-ERK1/2 in the mouse first maxillary molar. Recombinant lentivirus was constructed to overexpress DLK1 stably in hDPSCs. The cell viability and proliferation of hDPSCs were examined by CCK8 and EdU incorporation assay respectively. The odontoblastic differentiation of hDPSCs was determined by detection of ALPase activity assay, ALP and alizarin red staining and the expression of mineralization-related genes including ALP, DSPP and dental matrix protein. The mRNA and protein levels of DLK1 and p-ERK1/2 protein expression were detected. ERK inhibitor was used to test the differentiation effect of DLK1 on hDPSCs. RESULTS: Delta-like homologue 1 was highly expressed on the odontoblasts and dental pulp cells on the first maxillary molar; the expression of p-ERK1/2 is similar with the DLK1 in the same process. The expression level of DLK1 increased significantly after the odontoblastic induction of hDPSCs. DLK1 overexpression increased the proliferation ability of hDPSCs and inhibited odontoblastic differentiation of hDPSCs. The protein level of p-ERK1/2 significantly increased in hDPSCs/dlk1-oe group. ERK signalling pathway inhibitor reversed the odontoblastic differentiation effects of DLK1 on hDPSCs. CONCLUSIONS: The proliferation of hDPSCs was promoted after DLK1 overexpression. DLK1 inhibited the odontoblastic differentiation of hDPSCs, which maybe through ERK signalling pathway.


Assuntos
Diferenciação Celular , Proliferação de Células , Polpa Dentária/citologia , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteínas de Membrana/metabolismo , Odontoblastos/citologia , Odontoblastos/metabolismo , Células-Tronco/citologia , Animais , Proteínas de Ligação ao Cálcio , Células Cultivadas , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL
18.
Mater Sci Eng C Mater Biol Appl ; 72: 676-681, 2017 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-28024637

RESUMO

Magnesium and its alloys have attracted much attention as metallic biodegradable implants for their excellent biocompatibility and mechanical properties. However, magnesium has a poor corrosion resistance, causing its rapid degrading in vivo via an electrochemical reaction, which has become a major obstacle to their applications in implants. In this work, CaP coating was successfully coated on the ZK60 magnesium alloys by a simple hydrothermal deposition method. The mechanisms of the hydrothermal reactions of CaP coatings on Mg substrate are described in details. The effect of Ca/P ratio in the hydrothermal solution on the phase composition, microstructure and biodegradation properties of CaP coatings on ZK60 alloys was investigated by varying the Ca/P ratio from 0.83 to 4.18. The morphology of the CaP coating changed significantly with the Ca/P ratio. Biodegradation behavior of the CaP coating magnesium was characterized by anodic polarization and immersion tests in a simulated body fluid. It is revealed that the corrosion resistance of ZK60 magnesium alloy was greatly improved with the biomimetic CaP coatings, and the ZK60 alloy with CaP coating deposited at Ca/P ratio of 1.67 has the best corrosion resistance, which indicates that the CaP coatings are promising for improving the biodegradation properties of Mg-based orthopedic implants and devices.


Assuntos
Ligas/química , Cálcio/química , Materiais Revestidos Biocompatíveis/química , Fósforo/química , Corrosão , Técnicas Eletroquímicas , Microscopia Eletrônica de Varredura , Propriedades de Superfície , Difração de Raios X
19.
Curr Stem Cell Res Ther ; 11(3): 188-96, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-25882852

RESUMO

In recent years, beneficial effects of various ligands of three peroxisome-proliferator-activated receptor (PPAR) isoforms (α, ß, γ) have been reported in neurodegenerative diseases through delaying the onset and progression of diseases, reducing lesion size and improving functional recovery. Neural stem cells (NSCs) are assumed as a promising strategy for the treatment of human neurodegenerative diseases. PPARs are supposed to be one group of the key regulators of fate decisions in neural stem cells during development and adulthood, through their impact on the target genes involving cell proliferation, death and differentiation. The neuroprotective role of PPARs is suggested to be closely associated with the inflammation control and regenerative function of NSCs. Nevertheless, the molecular mechanisms remain to be elucidated. Here, we review the current knowledge about the beneficial role of PPARs in NSC development and neurogenesis and attempt to discuss the underlying mechanisms.


Assuntos
Diferenciação Celular , Proliferação de Células , Células-Tronco Neurais/citologia , Neurogênese/fisiologia , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Humanos , Doenças Neurodegenerativas/terapia , Fármacos Neuroprotetores
20.
Nephrology (Carlton) ; 21(3): 200-8, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26246161

RESUMO

AIM: Maternal dietary protein restriction reduces nephron number in offspring and increases the risk of cardiovascular and chronic kidney diseases. Perlecan is the major basement membrane/extracellular matrix heparan sulfate proteoglycan (HSPG) that plays a crucial role in nephron formation. This study was to determine whether maternal dietary protein restriction during pregnancy leads to an abnormal perlecan expression pattern during kidney development and a correlation with aberrant cell proliferation and apoptosis. METHODS: Pregnant Sprague-Dawley rats were divided into two groups, maintained on either a low-protein diet (MLP group) or a normal-protein diet (MNP group). Kidneys were dissected from embryos of different kidney development stages. Real-time PCR and immunohistochemistry were performed to detect the transcript level of rHSPG2, the coding gene of perlecan, and its protein expression pattern. Apoptosis and proliferation cell were detected by TUNEL system and Ki67 marker. RESULTS: Embryonic weights and nephron number were significantly affected by maternal low protein diets. The transcript level of rHSPG2 in the MLP group was significantly lower at embryonic day 18 and the neonatal period. Immunohistochemistry study was consistent with the RT-PCR results. The proliferation level of the MLP group was significantly lower than the MNP group at E18 and more apoptotic cells was detected in MLP newborn. CONCLUSION: Maternal protein restriction reduced the expression of perlecan and lead aberrant cell proliferation and apoptosis during mid-metanephrogenesis in offspring. This data may provide new evidence to understand the mechanism of reduced nephron number due to maternal protein restriction and enlighten solution.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Dieta com Restrição de Proteínas , Proteoglicanas de Heparan Sulfato/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Néfrons/metabolismo , Animais , Apoptose , Proliferação de Células , Regulação para Baixo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Idade Gestacional , Proteoglicanas de Heparan Sulfato/genética , Imuno-Histoquímica , Antígeno Ki-67/metabolismo , Néfrons/embriologia , Estado Nutricional , Organogênese , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real
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