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Animals adapt to environmental challenges with long-term changes at the behavioral, circuit, cellular, and synaptic levels which often require new protein synthesis. The discovery of reversible N6-methyladenosine (m 6 A) modifications of mRNA has revealed an important layer of post-transcriptional regulation which affects almost every phase of mRNA metabolism and therefore translational control. Many in vitro and in vivo studies have demonstrated the significant role of m 6 A in cell differentiation and survival, but its role in adult neurons is understudied. We used cell-type specific gene deletion of Mettl14, which encodes one of the subunits of the m 6 A methyltransferase, and Ythdf1 , which encodes one of the cytoplasmic m 6 A reader proteins, in dopamine D1 receptor expressing or D2 receptor expressing neurons. Mettl14 or Ythdf1 deficiency blunted responses to environmental challenges at the behavioral, cellular, and molecular levels. In three different behavioral paradigms, gene deletion of either Mettl14 or Ythdf1 in D1 neurons impaired D1-dependent learning, whereas gene deletion of either Mettl14 or Ythdf1 in D2 neurons impaired D2-dependent learning. At the cellular level, modulation of D1 and D2 neuron firing in response to changes in environments was blunted in all three behavioral paradigms in mutant mice. Ythdf1 deletion resembled impairment caused by Mettl14 deletion in a cell type-specific manner, suggesting YTHDF1 is the main mediator of the functional consequences of m 6 A mRNA methylation in the striatum. At the molecular level, while striatal neurons in control mice responded to elevated cAMP by increasing de novo protein synthesis, striatal neurons in Ythdf1 knockout mice didn't. Finally, boosting dopamine release by cocaine drastically increased YTHDF1 binding to many mRNA targets in the striatum, especially those that encode structural proteins, suggesting the initiation of long-term neuronal and/or synaptic structural changes. While the m6A-YTHDF1 pathway has similar functional significance at cellular level, its cell type specific deficiency in D1 and D2 neurons often resulted in contrasting behavioral phenotypes, allowing us to cleanly dissociate the opposing yet cooperative roles of D1 and D2 neurons.
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OBJECTIVE: To investigate the clinical efficacy of percutaneous screw fixation combined with minimally invasive transpedicular bone grafting and non-bone grafting in the treatment of thoracolumbar fractures. METHODS: From Janury 2021 to June 2022, 40 patients with thoracolumbar fracture were divided into the experimental group and the control group. There were 26 patients in the experimental group, including 21 males and 5 females with an aberage age of (47.3±12.3) years old, who underwent percutaneous pedicle screw fixation combined with transpedicular autogenous bone grafting. In the control group, 14 patients received percutaneous pedicle screw fixation only. including 7 makes and 7 females with an average age of (50.2±11.2) years old. The operative time, intraoperative blood loss, anterior height ratio of injured vertebrae, Cobb angle, visual analogue score (VAS), MacNab scores, loosening or broken of the implants. were compared and analyzed. RESULTS: There was no significant difference in operation time, intraoperative blood loss, VAS and anterior height ratio of injured vertebrae between the two groups. Compared with the preoperative results, VAS and anterior height ratio of injured vertebrae were improved statistically(P<0.05). For Cobb angle of injured vertebra, there was no significant difference between the two groups before surgery (P=0.766). While at 1 week, 3 months and 12 months after surgery, there were statistically differences between the two groups (P values were 0.042, 0.007 and 0.039, respectively). The Cobb angle of injured vertebrae one year after operation was statistically decreased in both groups compared with that before surgery (P<0.001). One year after surgery, the excellent and good rate of Macnab scores was 96.15% in the experimental group and 92.86% in the control group, and there was no statistical differences between the two groups (P=0.648). There was one patient in the control group suffering superficial wound infection on the third day, which was cured by dressing change and anti-infection treatment. There were no postoperative screw loosening and broken in both groups. CONCLUSION: The two surgical methods have the advantages of less trauma, less pain and quicker recovery, which can restore the height of the injured vertebra, reconstruct the spinal sequence and reduce the fracture of the vertebral body. Transpedicular autogenous bone grafting can increase the stability of the fractured vertebra and maintain the height of the vertebra better after surgery, thus reducing the possibility of complications such as kyphosis, screw loosening and broken.
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Tornillos Pediculares , Fracturas de la Columna Vertebral , Masculino , Femenino , Humanos , Adulto , Persona de Mediana Edad , Trasplante Óseo , Pérdida de Sangre Quirúrgica , Vértebras Lumbares/cirugía , Vértebras Lumbares/lesiones , Vértebras Torácicas/cirugía , Vértebras Torácicas/lesiones , Fijación Interna de Fracturas/métodos , Fracturas de la Columna Vertebral/cirugía , Resultado del Tratamiento , Estudios RetrospectivosRESUMEN
Pyridox(am)ine 5 ' -phosphate oxidase (PNPO) catalyzes the rate-limiting step in the synthesis of pyridoxal 5 ' -phosphate (PLP), the active form of vitamin B6 required for the synthesis of neurotransmitters gamma-aminobutyric acid (GABA) and the monoamines. Pathogenic variants in PNPO have been increasingly identified in patients with neonatal epileptic encephalopathy and early-onset epilepsy. These patients often exhibit different types of seizures and variable comorbidities. Recently, the PNPO gene has also been implicated in epilepsy in adults. It is unclear how these phenotypic variations are linked to specific PNPO alleles and to what degree diet can modify their expression. Using CRISPR-Cas9, we generated four knock-in Drosophila alleles, hWT , hR116Q , hD33V , and hR95H , in which the endogenous Drosophila PNPO was replaced by wild-type human PNPO complementary DNA (cDNA) and three epilepsy-associated variants. We found that these knock-in flies exhibited a wide range of phenotypes, including developmental impairments, abnormal locomotor activities, spontaneous seizures, and shortened life span. These phenotypes are allele dependent, varying with the known biochemical severity of these mutations and our characterized molecular defects. We also showed that diet treatments further diversified the phenotypes among alleles, and PLP supplementation at larval and adult stages prevented developmental impairments and seizures in adult flies, respectively. Furthermore, we found that hR95H had a significant dominant-negative effect, rendering heterozygous flies susceptible to seizures and premature death. Together, these results provide biological bases for the various phenotypes resulting from multifunction of PNPO, specific molecular and/or genetic properties of each PNPO variant, and differential allele-diet interactions.
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Alelos , Dieta , Epilepsia/genética , Fenotipo , Piridoxaminafosfato Oxidasa/genética , Vitamina B 6/metabolismo , Secuencia de Aminoácidos , Animales , Drosophila melanogaster , Humanos , Piridoxaminafosfato Oxidasa/química , Homología de Secuencia de AminoácidoRESUMEN
Distant hybridization usually leads to female sterility of the hybrid but the mechanism behind this is poorly understood. Complete pistil abortion but normal male fertility was shown by one Brassica napus-Orychophragmus violaceus monosomic alien addition line (MA, AACC + 1 IO, 2n = 39) produced previously. To study the effect of a single O. violaceus chromosome addition on pistil development in different genetic backgrounds, hybrids between the MA and B. carinata (BBCC), B. juncea (AABB), and two synthetic hexaploids (AABBCC) were firstly produced in this study which show complete female sterility. A microspore culture was further performed to produce the haploid monosomic alien addition line (HMA, AC + 1 IO, 2n = 20) and disomic addition line (DA, AACC + 2 IO, 2n = 40) together with haploid (H, AC, 2n = 19) and double haploid (DH, AACC, 2n = 38) plants of B. napus from MA to investigate the dosage effect of the alien O. violaceus chromosome on pistil development and gene expression. Compared to MA, the development of the pistils of DA and HMA was completely or partially recovered, in which the pistils could swell and elongate to a normal shape after open pollination, although no seeds were produced. Comparative RNA-seq analyses revealed that the numbers of the differentially expressed genes (DEGs) were significantly different, dosage-dependent, and consistent with the phenotypic difference in pairwise comparisons of HMA vs. H, DA vs. DH, MA vs. DH, MA vs. DA, and MA vs. HMA. The gene ontology (GO) enrichment analysis of DEGs showed that a number of genes involved in the development of the gynoecium, embryo sac, ovule, and integuments. Particularly, several common DEGs for pistil development shared in HMA vs. H and DA vs. DH showed functions in genotoxic stress response, auxin transport, and signaling and adaxial/abaxial axis specification. The results provided updated information for the molecular mechanisms behind the gynoecium development of B. napus responding to the dosage of alien O. violaceus chromosomes.
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BACKGROUND AND AIM: Acute treatment of young patients with proximal femoral fracture (PFF) remains a challenge for trauma surgeons due to major fracture displacement and heavy pain in clinical practice. Traditional methods have a variety of intrinsic defaults and cannot successfully manage the requirements of young patients. Benefiting from our anatomic research, we explored a new method of external fixation for this specific trauma and evaluated its feasibility and clinical outcomes. MATERIAL AND METHODS: Twenty-three young multiple-trauma patients with PFF were included in this study. Surgical treatment was applied using an external fixator via the anterior inferior iliac spine (AIIS). Electronic patient records, surgical characteristics, clinical outcomes, and complications were reviewed for each patient. RESULTS: The mean surgical time was 30.3 ± 7.3 min. The mean blood loss was 25.3 ± 10.8 mL. No iatrogenic nerve palsy, pin tract infection, failure of external fixation, or bedsores were observed. The postoperative visual analog scale score was significantly lower than the preoperative score (P < 0.01). The mean fracture reduction rate of the femur was 58.1 ± 17.0%, and the mean degree of reduction was 13.5 ± 6.9°. The mean external fixation time was 7.6 ± 4.0 days and intramedullary nailing was performed. The mean hospital, follow-up, and healing times were 28.7 ± 8.7 days, 23.5 ± 7.9 months, and 22.8 ± 5.7 weeks, respectively. The Harris Hip Score indicated excellent or good results in 20 patients. CONCLUSION: Collectively, the results of this study revealed that external fixation via the AIIS is a safe, rapid, and effective method for acute treatment of PFF in young patients.
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OBJECTIVE: The number of patients with metastatic bone tumors of the pelvis (MBTP) has increased, and the risk of metastasis and recurrence in the pelvic bones is difficult to assess. Therefore, we investigated the clinical features and oncological outcomes of patients with MBTP. METHODS: We analyzed the clinical features and oncological outcomes of MBTP in 72 patients (42 men, 30 women; mean age, 50.5 years) from 2008 to 2017. Recurrence in the pelvic bones and survival rates were analyzed with regard to patients' potential contributing factors. RESULTS: Enneking region I was the area most commonly containing MBTP (47.3%). Low- and high-grade tumors were identified in 40 and 32 patients, respectively. The most common pathological type was adenomatous carcinoma (34.7%), and the most common primary lesion was lung cancer (20.8%). The 3-year overall recurrence rate within the pelvic bones was 34.7%, and the 5-year overall survival rate was 29.2%. CONCLUSION: Patients with MBTP have a high risk of recurrence in the pelvic bones and poor survival after multimodal treatment. Pelvic recurrence might be affected by the metastatic involvement, tumor grade, surgical margins, and type of surgery, whereas the survival rate tends to be associated with the tumor grade.
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Neoplasias Óseas , Huesos Pélvicos , Neoplasias Pélvicas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Huesos Pélvicos/diagnóstico por imagen , Huesos Pélvicos/cirugía , Neoplasias Pélvicas/cirugía , Pelvis/diagnóstico por imagen , Estudios RetrospectivosRESUMEN
How to respond to starvation determines fitness. One prominent behavioral response is increased locomotor activities upon starvation, also known as Starvation-Induced Hyperactivity (SIH). SIH is paradoxical as it promotes food seeking but also increases energy expenditure. Despite its importance in fitness, the genetic contributions to SIH as a behavioral trait remains unexplored. Here, we examined SIH in the Drosophila melanogaster Genetic Reference Panel (DGRP) and performed genome-wide association studies. We identified 23 significant loci, corresponding to 14 genes, significantly associated with SIH in adult Drosophila. Gene enrichment analyses indicated that genes encoding ion channels and mRNA binding proteins (RBPs) were most enriched in SIH. We are especially interested in RBPs because they provide a potential mechanism to quickly change protein expression in response to environmental challenges. Using RNA interference, we validated the role of syp in regulating SIH. syp encodes Syncrip (Syp), an RBP. While ubiquitous knockdown of syp led to semi-lethality in adult flies, adult flies with neuron-specific syp knockdown were viable and exhibited decreased SIH. Using the Temporal and Regional Gene Expression Targeting (TARGET) system, we further confirmed the role of Syp in adult neurons in regulating SIH. To determine how syp is regulated by starvation, we performed RNA-seq using the heads of flies maintained under either food or starvation conditions. RNA-seq analyses revealed that syp was alternatively spliced under starvation while its expression level was unchanged. We further generated an alternatively-spliced-exon-specific knockout (KO) line and found that KO flies showed reduced SIH. Together, this study demonstrates a significant genetic contribution to SIH as a behavioral trait, identifies syp as a SIH gene, and highlights the significance of RBPs and post-transcriptional processes in the brain in regulating behavioral responses to starvation.
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Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Metabolismo Energético/genética , Estudio de Asociación del Genoma Completo/métodos , Proteínas de Unión al ARN/genética , Inanición , Alelos , Empalme Alternativo , Animales , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Drosophila melanogaster/fisiología , Regulación de la Expresión Génica , Frecuencia de los Genes , Locomoción/genética , Masculino , Neuronas/citología , Neuronas/metabolismo , Polimorfismo de Nucleótido Simple , Interferencia de ARN , Proteínas de Unión al ARN/metabolismo , RNA-Seq/métodosRESUMEN
Incorporation of azobenzene into the linker of bimetallic chiral salen TiIV catalysts allowed the photoswitchable arrangement of the two Ti(salen) units through cis/trans photoisomerization of azobenzene. The differently arranged Ti(salen) units changed their cooperative function to reflect the positional relationships, as a result, their efficiency as cooperative catalysts in asymmetric sulfoxidation could be readily controlled by light stimuli.
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Ionic liquid-functionalized amphiphilic Janus chiral salen TiIV catalysts were prepared by partial hydrophobic modification of silica with a chiral salen TiIV complex through an imidazolium ionic liquid (IL) linker. By optimizing their hydrophobic/hydrophilic balance, the IL-functionalized JNP materials exhibited excellent interfacial activity, significantly accelerating asymmetric sulfoxidation in water through the formation of stable Pickering emulsions. Moreover, catalyst recovery was readily achieved using centrifugation.
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Intrachain TiIV-oxazoline complexation together with hydrophobic interaction triggered the self-folding of an oxazoline-containing single polymeric chain in water. The formed TiIV-folded single-chain polymeric nanoparticles (SCPNs) acted as metalloenzyme-mimetic catalysts in asymmetric sulfoxidation in water owing to their organized, compartmentalized structure, effective site isolation, and also secondary coordination sphere provided by a copolymer backbone. In addition, they also could be facilely recovered for reuse by simple thermo-controlled separation.
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Objective To evaluate the clinical application of the minimally invasive modified pedicle screw-rod fixator for unstable pelvic ring injuries, including its feasibility, merits, and limitations. Methods Twenty-three patients (13 males, 10 females; average age, 36.3 years) with unstable pelvic ring injuries underwent anterior fixation using a modified pedicle screw-rod fixator with or without posterior fixation using a transiliac internal fixator. The clinical findings were assessed using Majeed scores. The quality of reduction was evaluated using the Matta criteria. Results Clinical results at 1 year postoperatively were excellent in 14 patients, good in 7, and fair in 2. The two patients with fair results had intermittent pain at the sacroiliac joint because of the posterior implant. One woman complained of persistent pain at the pubic tubercle during sexual intercourse. Iatrogenic neuropraxia of the unilateral lateral femoral cutaneous nerve occurred in three patients. Unilateral femoral nerve palsy occurred in one patient. The quality of fracture reduction was excellent in 12 patients, good in 8, and fair in 3. Heterotopic ossification occurred in eight patients; all were asymptomatic. Conclusions Minimally invasive modified pedicle screw-rod fixation is an effective alternative treatment for pelvic ring injuries.
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Fijación Interna de Fracturas/métodos , Fracturas Óseas/cirugía , Fijadores Internos , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Traumatismos de los Nervios Periféricos/cirugía , Adulto , Femenino , Nervio Femoral/lesiones , Nervio Femoral/cirugía , Fijación Interna de Fracturas/instrumentación , Fracturas Óseas/patología , Humanos , Masculino , Persona de Mediana Edad , Tornillos Pediculares , Huesos Pélvicos/lesiones , Huesos Pélvicos/inervación , Huesos Pélvicos/cirugía , Traumatismos de los Nervios Periféricos/patologíaRESUMEN
Oilseed rape (Brassica napus L.), which has yellow flowers, is both an important oil crop and a traditional tourism resource in China, whereas the Orychophragmus violaceus, which has purple flowers, likely possesses a candidate gene or genes to alter the flower colour of oilseed rape. A previously established B. napus line has a particular pair of O. violaceus chromosomes (M4) and exhibits slightly red petals. In this study, the transcriptomic analysis of M4, B. napus (H3), and O. violaceus with purple petals (OvP) and with white petals (OvW) revealed that most anthocyanin biosynthesis genes were up-regulated in both M4 and OvP. Read assembly and sequence alignment identified a homolog of AtPAP2 in M4, which produced the O. violaceus transcript (OvPAP2). The overexpression of OvPAP2 via the CaMV35S promoter in Arabidopsis thaliana led to different levels of anthocyanin accumulation in most organs, including the petals. However, the B. napus overexpression plants showed anthocyanin accumulation primarily in the anthers, but not the petals. However, when OvPAP2 was driven by the petal-specific promoter XY355, the transgenic B. napus plants produced red anthers and red petals. The results of metabolomic experiments showed that specific anthocyanins accumulated to high levels in the red petals. This study illustrates the feasibility of producing red-flowered oilseed rape, thereby enhancing its ornamental value, via the ectopic expression of the OvPAP2 gene. Moreover, the practical application of this study for insect pest management in the crop is discussed.
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Brassica napus/metabolismo , Flores/metabolismo , Antocianinas/metabolismo , Brassica napus/genética , Expresión Génica Ectópica/genética , Expresión Génica Ectópica/fisiología , Flores/genética , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismoRESUMEN
BACKGROUND: Unstable pelvic ring injuries often involve high mortality and morbidity. This study was aimed to evaluate the modified minimally invasive pedicle screw-rod fixation for anterior pelvic ring injuries, in the respects of its feasibility, merits, and limitations. METHODS: Twenty-three patients with unstable pelvic ring injuries underwent the modified anterior pedicle screw-rod fixation, with or without posterior fixation. The clinical outcomes were assessed using Majeed scores, and the quality of reduction was evaluated according to the criteria of Matta. RESULTS: Majeed scores showed that the clinical outcomes at postoperatively 1 year were excellent in 14 patients, good in 7, and fair in 2. One woman complained of persistent pain at the pubic tubercle during sexual intercourse. Iatrogenic neuropraxia of the unilateral lateral femoral cutaneous nerve occurred in 3 patients. Unilateral femoral nerve palsy occurred in 1 patient. The reduction was found to be excellent in 12 patients, good in 8, and fair in 3. Heterotopic ossification occurred in 8 patients, all being asymptomatic. CONCLUSIONS: The modified pedicle screw-rod fixation with the minimally invasive technique offered an effective alternative for unstable anterior pelvic ring injuries.
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Fijación Interna de Fracturas/métodos , Fracturas Óseas/cirugía , Tornillos Pediculares , Huesos Pélvicos/lesiones , Adulto , Femenino , Estudios de Seguimiento , Fijación Interna de Fracturas/efectos adversos , Fijación Interna de Fracturas/instrumentación , Fracturas Óseas/diagnóstico por imagen , Humanos , Masculino , Persona de Mediana Edad , Procedimientos Quirúrgicos Mínimamente Invasivos/efectos adversos , Procedimientos Quirúrgicos Mínimamente Invasivos/instrumentación , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Huesos Pélvicos/diagnóstico por imagen , Huesos Pélvicos/cirugía , Cuidados Posoperatorios/métodos , Radiografía , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Adulto JovenRESUMEN
Autophagy has been shown to antagonize the development of Lumbar disc degeneration (LDD), whereas the molecular regulation of autophagy is unknown. We recently reported a potential role of Insulin-like growth factor 1 receptor (IGF1R)/phosphatidylinositol-3 kinase (PI3k)/Akt signaling in the initiation and progression of LDD. Here, we studied the effects of IGF1R signaling on disc cell autophagy. We showed a correction of activation of IGF1R and disc cell autophagy in the resected discs in LDD patients. In vitro, activation of IGF1R signaling antagonized the decreases in cell viability of human disc cells, HNPSV, through suppression of apoptosis and enhancement of autophagy. Suppression of IGF1R signaling or inhibition of autophagy abolished the effects of activation of IGF1R signaling on disc cell survival upon compression. Together, our data suggest that activation of IGF1R may antagonize LDD, at least partially through enhanced autophagy.
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A potential role of Insulin-like growth factor 1 (IGF1) receptor/phosphatidylinositol-3 kinase (PI3k)/Akt signaling in the initiation and progression of Lumbar disc degeneration (LDD) has been recently reported. However, the regulation of IGF1 receptor (IGF1R) at post-transcriptional levels in the development of LDD remains unknown. Here, we studied the effects of microRNA-4458 on the expression of IGF1R. We examined the IGF1R levels and microRNA-4458 (miR-4458) levels in the resected LDD discs, compared with the traumatized, non-LDD discs. We analyzed the binding of miR-4458 to the 3'-UTR of IGF1R mRNA and its effects on IGF1R translation by bioinformatics analysis and by luciferase-reporter assay, respectively. We modified miR-4458 levels in a human nucleus pulposus SV40 cell line (HNPSV), and examined the effects of miR-4458 on the expression of IGF1R and Akt, as well as their phosphorylation. We found that the levels of miR-4458 were significantly higher and the levels of IGF1R were significantly lower in LDD discs, compared with the control non-LDD discs. The levels of IGF1R inversely correlated with the levels of miR-4458 in LDD discs. Moreover, miR-4458 was found to bind to the 3'-UTR of IGF1R mRNA to prevent its translation. In miR-4458-modified HNPSV cells, we found that miR-4458 decreased both total IGF1R and phosphorylated IGF1R, resulting in deceases in phosphorylated Akt. Thus, these data suggest that miR-4458 may suppress PI3k/Akt signaling pathway through 3'-UTR-inhibtion of IGF1R mRNA to promote development of LDD.
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BACKGROUND/AIMS: The pathogenesis of Lumbar disc degeneration (LDD) has been extensively studied in the past. In particular, a role of matrix metalloproteinase 3 (MMP3) in the disease initiation and progression has been recently reported. However, an involvement of Insulin-like growth factor 1 (IGF-I)-stimulated phosphatidylinositol-3 kinase (PI3k) / Akt signaling pathway-mediated control of MMP3 in LDD has not been acknowledged. METHODS: We examined the serum IGF-1 levels and activation of the receptor for IGF-1 (IGF-1R) in resected discs in patients with LDD, compared to the fractured discs from traumatized, non-LDD subjects as a control. We analyzed the effects of IGF-1 on the activation of IGF-1R, Akt and MMP3 in a human nucleus pulposus SV40 cell line (HNPSV). We transfected HNPSV cells with a constitutive nuclear FoxO1, and analyzed its effect on the activation of IGF-1R, Akt and MMP3. RESULTS: LDD patients had significantly lower levels of serum IGF-1, and LDD discs had significantly lower levels of activated IGF-1R. IGF-1 induced phosphorylation of IGF-1R, and then phosphorylation of its downstream factor Akt in the HNPSV cells, resulting in significantly inhibition of MMP3. Further, FoxO1 nuclear retention completely abolished the inhibitory effects of IGF-1 on MMP3 in HNPSV cells. CONCLUSION: Together, IGF-1/Akt/FoxO1/MMP3 regulatory machinery may control the development of LDD.
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Factor I del Crecimiento Similar a la Insulina/metabolismo , Degeneración del Disco Intervertebral/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/fisiología , Línea Celular , Proteína Forkhead Box O1 , Factores de Transcripción Forkhead/metabolismo , Humanos , Metaloproteinasa 3 de la Matriz/metabolismo , Fosforilación/fisiología , Receptor IGF Tipo 1/metabolismoRESUMEN
BACKGROUND: The gynoecium is one of the most complex organs of angiosperms specialized for seed production and dispersal, but only several genes important for ovule or embryo sac development were identified by using female sterile mutants. The female sterility in oilseed rape (Brassica napus) was before found to be related with one alien chromosome from another crucifer Orychophragmus violaceus. Herein, the developmental anatomy and comparative transcript profiling (RNA-seq) for the female sterility were performed to reveal the genes and possible metabolic pathways behind the formation of the damaged gynoecium. RESULTS: The ovules in the female sterile Brassica napus with two copies of the alien chromosomes (S1) initiated only one short integument primordium which underwent no further development and the female gametophyte development was blocked after the tetrad stage but before megagametogenesis initiation. Using Brassica_ 95k_ unigene as the reference genome, a total of 28,065 and 27,653 unigenes were identified to be transcribed in S1 and donor B. napus (H3), respectively. Further comparison of the transcript abundance between S1 and H3 revealed that 4540 unigenes showed more than two fold expression differences. Gene ontology and pathway enrichment analysis of the Differentially Expressed Genes (DEGs) showed that a number of important genes and metabolism pathways were involved in the development of gynoecium, embryo sac, ovule, integuments as well as the interactions between pollen and pistil. CONCLUSIONS: DEGs for the ovule development were detected to function in the metabolism pathways regulating brassinosteroid (BR) biosynthesis, adaxial/abaxial axis specification, auxin transport and signaling. A model was proposed to show the possible roles and interactions of these pathways for the sterile gynoecium development. The results provided new information for the molecular mechanisms behind the gynoecium development at early stage in B. napus.