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OBJECTIVE: In the present study, we evaluated and compared the outcomes of transarterial embolization with those of percutaneous contrast-enhanced ultrasound-guided transabdominal sac embolization (PUSE) for type II endoleaks (T2ELs). METHODS: A retrospective review was conducted of consecutive patients who had undergone T2EL embolization between January 2015 and December 2020 at our center. The cohort was divided into two groups according to the embolization approach: PUSE vs transarterial. Freedom from aneurysm growth, safety, immediate technical success, freedom from persistent T2ELs, and the repeat embolization rate were assessed. RESULTS: A total of 25 patients and 28 embolization procedures (PUSE, n = 16; transarterial embolization, n = 12) were examined. Both the fluoroscopic time (13.3 ± 3.2 minutes vs 35.0 ± 7.0 minutes; P < .001) and the procedural time (84.9 ± 8.4 minutes vs 117.1 ± 14.8 minutes; P < .001) were significantly shorter in the PUSE group than in the transarterial group. After the embolization procedure, the patients were followed up for a mean duration of 24.7 ± 14.9 months for the PUSE group and 35.9 ± 21.1 months for the transarterial group (P = .1323). Five patients in the transarterial group had undergone unsuccessful embolization, with success in 7 of the 12 patients in the transarterial group and all 16 patients in the PUSE group (P = .0081). Failure had resulted from failed transarterial access or a recurrent T2EL. Three of the five patients had undergone subsequent PUSE during follow-up. No patient in the PUSE group had experienced sac expansion compared with four patients in the transarterial group (P = .0242). Similarly, no patient in the PUSE group had developed a newly discovered T2EL vs four patients in the transarterial group (P = .0242). Thus, the outcomes were markedly better for the PUSE group than were those for the transarterial group. A major procedure-related complication (abdominal abscess) occurred in one patient in the transarterial group. CONCLUSIONS: PUSE is safe and effective for managing T2ELs. It yields better outcomes in terms of preventing aneurysm growth, decreasing the incidence of repeat embolization and complications, minimizing the recurrence of T2ELs, and reducing the fluoroscopic and procedural times. We, thus, regard it as the preferred approach for the management of T2ELs.
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Aneurisma da Aorta Abdominal , Implante de Prótese Vascular , Embolização Terapêutica , Procedimentos Endovasculares , Aneurisma da Aorta Abdominal/complicações , Aneurisma da Aorta Abdominal/diagnóstico por imagem , Aneurisma da Aorta Abdominal/cirurgia , Implante de Prótese Vascular/efeitos adversos , Embolização Terapêutica/efeitos adversos , Embolização Terapêutica/métodos , Endoleak/diagnóstico por imagem , Endoleak/etiologia , Endoleak/terapia , Procedimentos Endovasculares/efeitos adversos , Humanos , Estudos Retrospectivos , Fatores de Tempo , Resultado do Tratamento , Ultrassonografia de Intervenção/efeitos adversosRESUMO
Inflammation is a common situation during bone healing and is recognized to inhibit osteogenic differentiation and bone formation. However, the effect of inflammation on BMP-9-induced osteoblastic differentiation remains unclear. In the present study, we found that an inflammatory environment triggered by lipopolysaccharide (LPS) in vitro suppressed BMP-9-induced osteogenic differentiation. In addition, LPS decreased BMP-9-induced phosphorylation of Smad1/5/8 and showed obvious inhibitory effects on BMP-9-induced Smad signaling. We then confirmed that LPS and BMP-9 can activate p38MAPK and ERK1/2 signaling, and that LPS stimulation reduces BMP-9-induced Runx2 expression through the activation of p38MAPK and ERK1/2 signaling. Finally, we determined that blockade of MAPK signaling by specific inhibitors reverses the inhibitory effect of LPS on BMP-9-induced osteogenic differentiation, and that MAPK acts as a mediator of the negative regulatory role of LPS in BMP-9-induced activation of Smad signaling. Based on these results, we conclude that the LPS-mediated inflammatory environment inhibits BMP-9-induced osteogenic differentiation, via crosstalk between BMP/MAPK and Smad signaling. The elucidation of these mechanisms may hasten the development of new strategies and improve the osteoinductive efficacy of BMP-9 in the clinic, resulting in enhanced osteoblastic differentiation and bone formation.
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Diferenciação Celular/efeitos dos fármacos , Fator 2 de Diferenciação de Crescimento/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Osteoblastos/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Proteínas Smad/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Fator 2 de Diferenciação de Crescimento/metabolismo , Inflamação/induzido quimicamente , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismoRESUMO
Apolipophorin-III (ApoLp-III) acts in lipid transport, lipoprotein metabolism, and innate immunity in insects. In this study, an ApoLp-III gene of Antheraea pernyi pupae (Ap-ApoLp-III) was isolated and characterized. The full-length cDNA of Ap-ApoLp-III is 687 bp, including a 5'-untranslated region (UTR) of 40 bp, 3'-UTR of 86 bp and an open reading frame of 561 bp encoding a polypeptide of 186 amino acids that contains an Apolipophorin-III precursor domain (PF07464). The deduced Ap-apoLp-III protein sequence has 68, 59, and 23% identity with its orthologs of Manduca sexta, Bombyx mori, and Aedes aegypti, respectively. Phylogenetic analysis showed that the Ap-apoLp-III was close to that of Bombycoidea. qPCR analysis revealed that Ap-ApoLp-III expressed during the four developmental stages and in integument, fat body, and ovaries. After six types of microorganism infections, expression levels of the Ap-ApoLp-III gene were upregulated significantly at different time points compared with control. RNA interference (RNAi) of Ap-ApoLp-III showed that the expression of Ap-ApoLp-III was significantly downregulated using qPCR after injection of E. coli. We infer that the Ap-ApoLp-III gene acts in the innate immunity of A. pernyi.
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Apolipoproteínas/genética , Imunidade Inata , Mariposas/genética , Sequência de Aminoácidos , Animais , Apolipoproteínas/biossíntese , Apolipoproteínas/imunologia , Sequência de Bases , DNA Complementar , Feminino , Estágios do Ciclo de Vida , Masculino , Dados de Sequência Molecular , Mariposas/imunologia , Mariposas/microbiologia , Fases de Leitura Aberta , Filogenia , Interferência de RNA , Reação em Cadeia da Polimerase em Tempo Real , Regiões não TraduzidasRESUMO
The purpose of this study is to evaluate the efficacy of plate augmentation and hybrid bone grafting for treating atrophic nonunion of the femur with original intramedullary nail retained in situ.In this study, 36 patients with atrophic nonunion of the femur who underwent surgery using the technique of plate augmentation and a hybrid bone grafting while retaining the original intramedullary nail in situ in Xi'an Honghui Hospital from January 2019 to December 2021 were enrolled. 28 patients who met the inclusion and exclusion criteria were ultimately included in the study. These 28 patients, consisting of 20 males and 8 females with a mean age of 38 years, were evaluated based on factors such as operation time, intraoperative blood loss, the average hospitalization days. Additionally, the results and function of these patients were evaluated by union time, Wu's scores of limb function and incidence of serious complications.All 28 patients achieved bone union at the 12 month follow-up, with an average follow-up time of 14.6 ± 4.2 months.The average operation time was 68.3 ± 11.2 min, and the average intraoperative blood loss was 140 ± 22.6 ml. Patients were hospitalized for an average of 5.8 ± 1.1 days. Full clinical and radiological bone union was achieved on average at 5.1 ± 1.9 months. The mean value of Wu's scores at the 12 month follow-up was significantly higher than before the operation. Limb function was excellent in 27 patients and good in one patient at the 12 month follow-up. However, five patients experienced the lower limb vein thrombosis, including one deep vein thrombosis and four lower limb intermuscular vein thromboses. One patient had a superficial infections of the surgical incision site, while three patients reported pain and numbness where their iliac bone graft was extracted at the 12 month follow-up. The technique of plate augmentation and hybrid bone grafting, combined with retaining the original intramedullary nail in situ has been shown to be a safe, effective, simply and standardizable practice for treating atrophic femoral nonunion with an intact original IMN fixation.
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Fixação Intramedular de Fraturas , Fraturas não Consolidadas , Masculino , Feminino , Humanos , Adulto , Transplante Ósseo/métodos , Perda Sanguínea Cirúrgica , Fixação Intramedular de Fraturas/métodos , Pinos Ortopédicos/efeitos adversos , Fraturas não Consolidadas/cirurgia , Resultado do Tratamento , Fêmur/cirurgia , Extremidade Inferior , Estudos Retrospectivos , Consolidação da FraturaRESUMO
OBJECTIVES: Brucellar cervical epidural abscess (CEA) is a rare condition with potentially permanent neurological damage if left untreated. This study aims to define the clinical presentation of brucellar CEA and evaluate the outcome of surgical treatment, specifically decompression and fusion surgery. The findings will contribute to understanding whether all patients with brucellar CEA could benefit from this surgical intervention. METHODS: A retrospective study on brucellar spondylitis was conducted at the First Hospital of Jilin University from August 2018 to August 2022. During this period, a total of 37 patients were diagnosed with brucellar spondylitis at the hospital. Out of the 37 cases, six patients (16.2%) were confirmed to have CEA through cervical magnetic resonance imaging examination and serology test results.. RESULTS: Six patients were diagnosed with brucellar CEA (16.2%), of whom 5 successfully underwent anterior cervical decompression and fusion surgery. One patient had a large prevertebral abscess that could only be drained. In combination with effective antibiotic therapy, the clinical performance of the 5 patients who underwent surgery improved after the surgery. The remaining one patient required delayed surgery due to instability of the cervical spine. The follow-up period of all the 6 patients was 6 months. CONCLUSIONS: Brucellosis should be considered as a potential cause of CEA, especially in endemic areas. Timely detection and effective management of this condition are crucial in order to minimize the associated morbidity and mortality. For patients with detectable brucellar CEA, we recommend decompression and fusion surgery.
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Brucella , Brucelose , Abscesso Epidural , Espondilite , Humanos , Abscesso Epidural/diagnóstico por imagem , Abscesso Epidural/cirurgia , Abscesso Epidural/tratamento farmacológico , Estudos Retrospectivos , Brucelose/complicações , Espondilite/complicações , Imageamento por Ressonância MagnéticaRESUMO
Precise targeting of specific regions within the central nervous system (CNS) is crucial for both scientific research and gene therapy in the context of brain diseases. Adeno-associated virus 13 (AAV13) is known for its restricted diffusion range within the CNS, making it an ideal choice for precise labeling and administration within small brain regions. However, AAV13 mediates relatively low expression of target genes. Here, we introduced specifically engineered modifications to the AAV13 capsid protein to enhance its transduction efficiency. We first constructed AAV13-YF by mutating tyrosine to phenylalanine on the surface of the AAV13 capsid. We then inserted the 7m8 peptide, known to enhance cell transduction, into positions 587/588 and 585/586 of the AAV13 capsid, resulting in two distinct variants named AAV13-587-7m8 and AAV13-585-7m8, respectively. We found that AAV13-YF exhibited superior in vitro infectivity in HEK293T cells compared to AAV13, while AAV13-587-7m8 and AAV13-585-7m8 showed enhanced CNS infection capabilities in C57BL/6 mice, with AAV13-587-7m8 infection retaining a limited spread range. These modified AAV13 variants hold promising potential for applications in gene therapy and neuroscience research.
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Dependovirus , Camundongos Endogâmicos C57BL , Dependovirus/genética , Animais , Humanos , Camundongos , Células HEK293 , Transdução Genética , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismoRESUMO
The smooth surface of the metallic nanostructure is essential for the propagation of surface plasmon polaritons. In this paper, we present a novel method to fabricate the metallic nanopatterns with ultra-smooth surface on various substrates. By using a silica film as the sacrificial layer, we show that the prefabricated metallic nanopatterns produced by electron beam lithography and film deposition can be hydrolyzed and transferred onto a designated substrate. The ultra-smooth surface morphology of nanopatterns has been characterized and verified by scanning electron microscopy and atomic force microscopy. More importantly, we demonstrate that this method can successfully produce a variety of nanostructures with high product yield, even onto the uneven substrate. The results indicate that our proposed method is a promising and versatile means to fabricate multiplicate smooth metallic nanostructure on various substrates for the application of nanophotonic devices.
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Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Impressão Molecular/métodos , Refratometria/métodos , Elétrons , Teste de Materiais , Nanopartículas Metálicas/efeitos da radiação , Propriedades de Superfície/efeitos da radiaçãoRESUMO
Insects possess an innate immune system that responds to invading microorganisms. In this study, a subtractive cDNA library was constructed to screen for immune response-related genes in the fat bodies of Antheraea pernyi (Lepidoptera: Saturniidae) pupa challenged with Escherichia coli. Four hundred putative EST clones were identified by suppression subtractive hybridization (SSH), including 50 immune response-related genes, three cytoskeleton genes, eight cell cycle and apoptosis genes, five respiration and energy metabolism genes, five transport genes, 40 metabolism genes, ten stress response genes, four transcription and translation regulation genes and 77 unknown genes. To verify the reliability of the SSH data, the transcription of a set of randomly selected immune response-related genes were confirmed by semi-quantitative reverse transcription-PCR (RT-PCR) and real-time quantitative reverse transcription-PCR (qRT-PCR). These identified immune response-related genes provide insight into understanding the innate immunity in A. pernyi.
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Bombyx/imunologia , Genes de Insetos , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Bombyx/microbiologia , Escherichia coli/imunologia , Corpo Adiposo/metabolismo , Biblioteca Gênica , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de SequênciaRESUMO
As radiation therapy is increasingly utilized in the treatment of cancer, neuropathic pain (NP) is a common radiotherapy-related adverse effect and has a significant impact on clinical outcomes negatively. However, despite an improved understanding of neuropathic pain management, pain is often undertreated in patients with cancer. Herein, we reported two cases with radiotherapy-related neuropathic pain (RRNP) who presented a positive reaction to acupuncture. Patient 1 (a 73-year-old woman) with gynecologic cancer complained of burning and electric shock-like pain in the lower limb after radiotherapy. With the accepted combination of acupuncture and drugs, the pain was alleviated completely in 8 weeks. Patient 2 (a 64-year-old woman) accepted acupuncture in the absence of medication because of her inability to tolerate the adverse events of anticonvulsant drugs. She achieved remission of pain 4 weeks later. The results of this study showed that acupuncture might be promising for controlling the RRNP in patients with cancer, especially who were intolerant or unresponsive to medications.
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Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases. Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing, making these vectors powerful candidates for the dissection of synaptic inputs. Although several attenuated rabies viral vectors have been developed, their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors. To overcome these limitations, we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus. Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably: (1) N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro, and the efficiency was six times higher than that of rAAV9-Retro; (2) the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2-3 times higher than that of oG-mediated SAD-B19-ΔG; (3) CVS-N2c-ΔG could delivery modified genes for neural activity monitoring, and the time window during which this was maintained was 3 weeks; and (4) CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination. These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.
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OBJECTIVE: We investigated the safety and efficacy of the Numen coil compared with the Axium coil in the treatment of intracranial aneurysms. METHODS: Because CATCH (Coil Application Trial in China) is a prospective randomized controlled open-label noninferiority trial conducted in 10 centers across China, patients who fulfilled the inclusion and exclusion criteria were randomized 1:1 to either a test group (Numen) or a control group (Axium). The primary outcome was based on successful aneurysm occlusion at 6 months follow-up, whereas secondary outcomes included technical success, the recanalization and retreatment rates, and the rate of serious adverse events (SAEs) at 6 months and 12 months follow-up. RESULTS: Between August 2017 and December 2019, 350 patients presenting with 350 aneurysms were enrolled and randomized. Per-protocol analysis showed that the successful aneurysm occlusion rate at 6 months was 91.18% for the test group compared with 91.85% in the control group, with a difference of -0.68% (P = 0.8419), and the overall mortality during the 30-day follow-up period was 1.19% and 1.81% in the test and control group, respectively, showing no significant difference between the 2 groups (P = 0.6837), whereas the SAE incidence during the 12-month follow-up period was 12.50% and 17.47% in the test and control groups, respectively, which was not statistically significant (P = 0.2222). CONCLUSIONS: This trial showed that the Numen coil was noninferior to the Axium coil in terms of intracranial aneurysm embolization and can be considered as a safe and effective coil for treating patients with intracranial aneurysm in clinical practice.
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Aneurisma da Aorta Abdominal , Implante de Prótese Vascular , Embolização Terapêutica , Procedimentos Endovasculares , Aneurisma Intracraniano , Embolização Terapêutica/métodos , Procedimentos Endovasculares/métodos , Seguimentos , Humanos , Aneurisma Intracraniano/etiologia , Aneurisma Intracraniano/cirurgia , Estudos Prospectivos , Resultado do TratamentoRESUMO
Deep brachial artery aneurysms are extremely rare. The purpose of this article is to report a case of deep brachial artery aneurysm that was successfully treated by open surgery. A 76-year-old man presented with complaints of an asymptomatic pulsatile mass in the left axilla. A computed tomography angiography revealed a deep brachial artery aneurysm. The aneurysm was resected surgically, then the axillary artery was repaired, and the distal end of the deep brachial artery was ligated without vascular reconstruction. The patient had a good recovery with no complications, and the arterial pulses of the left upper extremity were normal.
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Aneurisma/cirurgia , Artéria Braquial/cirurgia , Procedimentos Cirúrgicos Vasculares , Idoso , Aneurisma/diagnóstico por imagem , Artéria Braquial/diagnóstico por imagem , Humanos , Ligadura , Masculino , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
The aim of this study was to investigate whether a moderate-intensity static magnetic field (SMF) can enhance the killing effect of adriamycin (ADM) on K562 cells, and to explore the effects of SMF combined with ADM on K562 cells. We analyzed the metabolic activity of cells, cell cycle distribution, DNA damage, change in cell ultrastructure, and P-glycoprotein (P-gp) expression after K562 cells were exposed continuously to a uniform 8.8 mT SMF for 12 h, with or without ADM. Our results showed that the SMF combined with ADM (25 ng/ml) significantly inhibited the metabolic activity of K562 cells (P < 0.05), while neither ADM nor the SMF alone affected the metabolic activity of these cells. Cell ultrastructure was altered in the SMF + ADM group. For example, cell membrane was depressed, some protuberances were observable, and vacuoles in the cytoplasm became larger. Cells were arrested at the G2/M phase and DNA damage increased after cells were treated with the SMF plus ADM. ADM also induced the P-gp expression. In contrast, in the SMF group and SMF + ADM group, the P-gp expression was decreased compared with the ADM group. Taken together, our results showed that the 8.8 mT SMF enhanced the cytotoxicity potency of ADM on K562 cells, and the decrease in P-gp expression may be one reason underlying this effect.
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Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Magnetismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Dano ao DNA , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Humanos , Células K562 , Organelas/efeitos dos fármacos , Organelas/ultraestruturaRESUMO
This study aimed to investigate the relationship between the incidence of deep vein thrombosis (DVT) during hospitalization and the energy of injury in tibial plateau fractures (TPFs). One hundred and forty patients were enrolled between September 1, 2014, and October 1, 2017. According to Schatzker's classification, they were classified into the low-energy (type I-III) and high-energy (type IV-VI) groups. For DVT evaluation, duplex ultrasonography was performed in the lower extremities preoperatively and postoperatively. The location and changes of DVT were recorded. All patients underwent mechanical and chemical thromboprophylaxis. The incidence of DVT in TPFs was 36.43% and 46.43% preoperatively and postoperatively, respectively. The DVT incidence was 31.75% (20/63) in the low-energy group and 40.26% (31/77) in the high-energy group preoperatively, and 44.44% (28/63) in the low-energy group and 48.05% (37/77) in the high-energy group postoperatively. There was no significant difference between the 2 groups preoperatively (P = .298) and postoperatively (P = .785). The days between operation and discharge (P = .016), blood loss during surgery (P = .016), and preoperative d-dimer level (P = .02) showed differences between the 2 groups. Additionally, 29 new thrombi (14 [48.28%] in the high-energy group and 15 [51.72%] in the low-energy group) appeared and 16 preoperative thrombi disappeared postoperatively. Despite mechanical and chemical thromboprophylaxis, the DVT risk in patients with TPFs remains high. Although the DVT incidence is not significantly different between high-energy and low-energy injuries, the occurrence of DVT should be carefully monitored.
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Fraturas da Tíbia/complicações , Trombose Venosa/etiologia , Adulto , Idoso , Feminino , Hospitalização , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Fraturas da Tíbia/cirurgia , Trombose Venosa/patologia , Adulto JovemRESUMO
The tectonic boundaries and geodynamic evolution of the South China Block are widely debated. While the community largely agrees on the occurrence of the collision between the Yangtze and Cathaysia Blocks, the lack of ultrahigh-pressure metamorphic rocks and obscurity of the boundary lead to inconsistencies among the abundant geological and geophysical data. We present three profiles that reveal the geoelectrical structure of eastern South China. Distinct conductive interfaces oriented NE-SW are identified in the geoelectrical lithosphere, which separate the region into six parts. To interpret our observations and resultant model, we develop and propose a mechanism of "microcontinent interaction". Our new model justifies the prior proposed models of 'block collision' and additionally proposes 'multi-terrane accretion-collision' to address the tectonic evolution.
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A novel scheme to correct aberration of each beam from the front-end to the target point in a thermal inertia laser (TIL) is presented. Each beam contains a deformable mirror (DM) with an aperture of 70 mm x 70 mm at the injection of the main amplifier and a Hartman-Shack (HS) sensor in a parameter diagnostic unit (PDU). A temporary HS sensor for measuring the static aberration of each beam with 1 Hz source is placed at the target point. The sensor will be removed from the target point during the main single shot, so we transfer the results measured at the target point to the sensors in the PDU. Dynamic aberration can also be measured by the HS sensor in the PDU during the single shot. In this way, we need not calibrate the aberration of the PDU, and aberration of each beam can be corrected by the DM with the HS sensor in the PDU. We demonstrate that with this scheme the divergence angle of the TIL pulses can be improved from 100 to less than 60 murad with a focal length of 2200 mm and beam size of 290 mm x 290 mm, which meets the requirement of a TIL.
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Epidermal growth factor-like repeats and discoidin I like domain 3 (EDIL3) is an integrin ligand which is implicated in bone metabolism and bone marrow myelopoiesis. Recently, myeloid derived suppressor cells (MDSCs) as osteoclast progenitor have been demonstrated in several kinds of cancers including breast cancer. In this paper we explored the association between tumor derived EDIL3 and MDSCs in a murine breast cancer model. Knockdown of EDIL3 in MDA-MB-231 breast cancer cells inhibited the expansion of tumor induced MDSCs in bone marrow. However, generation of bone marrow derived MDSCs in vitro was not affected by recombinant EDIL3. Osteoclastogenesis of MDSCs was dose-dependently inhibited by recombinant EDIL3 in vitro via binding to Mac-1 but not LFA-1. Moreover, in accordance with previous studies, our data showed that tumor derived EDIL3 was involved in tumor associated bone loss. The convoluted effects of EDIL3 on MDSCs compose a potential mechanism hired by tumor cells for perpetration approximately.
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BACKGROUND: U6 promoters have been used for single guide RNA (sgRNA) transcription in the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas9) genome editing system. However, no available U6 promoters have been identified in Aspergillus niger, which is an important industrial platform for organic acid and protein production. Two CRISPR/Cas9 systems established in A. niger have recourse to the RNA polymerase II promoter or in vitro transcription for sgRNA synthesis, but these approaches generally increase cloning efforts and genetic manipulation. The validation of functional RNA polymerase II promoters is therefore an urgent need for A. niger. RESULTS: Here, we developed a novel CRISPR/Cas9 system in A. niger for sgRNA expression, based on one endogenous U6 promoter and two heterologous U6 promoters. The three tested U6 promoters enabled sgRNA transcription and the disruption of the polyketide synthase albA gene in A. niger. Furthermore, this system enabled highly efficient gene insertion at the targeted genome loci in A. niger using donor DNAs with homologous arms as short as 40-bp. CONCLUSIONS: This study demonstrated that both heterologous and endogenous U6 promoters were functional for sgRNA expression in A. niger. Based on this result, a novel and simple CRISPR/Cas9 toolbox was established in A. niger, that will benefit future gene functional analysis and genome editing.
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The objective of the present study was to investigate the effects of leptin addition in in vitro maturation (IVM) medium on meiotic maturation of oocytes and preimplantation development of parthenogenetic and cloned embryos in pigs. In experiment 1, oocytes were matured in North Carolina State University 23 (NCSU-23) medium supplemented with various concentrations of leptin: 0, 1, 10 and 100 ng/ml. IVM medium added with 10 or 100 ng/ml leptin significantly increased the rate of oocytes reaching metaphase II compared to the control (76.8% and 73.8% versus 61.7%). In experiment 2, the influence of the timing of leptin addition in IVM medium on meiotic maturation of porcine oocytes was assessed, and maximum maturation rate of oocytes developing to metaphase II was achieved when supplemented during the first half (0-22 h), the latter half (22-44 h) or the entire maturation period (0-44 h) compared to the control (80.5%, 84.7% and 78.1% versus 70.4%). In experiment 3, leptin strikingly increased the blastocyst rate of parthenogenetic embryos at the concentration of 10 ng/ml (37.5% versus 21.7%) and this increase was independent of the addition timing (0-44, 0-22, 22-44 h) compared to the control (32.5%, 34.6% and 31.5% versus 16.2%). Moreover, total cell number per blastocyst of parthenogenetic embryos was obviously increased in the 10 and 100 ng/ml leptin treatments as compared with the control (36, 38 versus 28). In experiment 4, 10 ng/ml leptin treatment significantly increased the rate of cleavage (72% versus 56%) of cloned embryos. Meanwhile, the rate of blastocyst formation was also improved although no significant difference was found (12.8% versus 7.1%). Collectively, our results indicate that leptin supplementation in IVM medium may be beneficial not only for developmental potential of oocytes but for subsequent developmental competence of embryos produced by parthenogenetic activation and the cleavage of embryos derived by somatic cell nuclear transfer (SCNT).
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Técnicas de Cultura de Células , Clonagem de Organismos , Desenvolvimento Embrionário/efeitos dos fármacos , Leptina/farmacologia , Meiose/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Oogênese/efeitos dos fármacos , Partenogênese , Suínos/embriologia , Animais , Meios de Cultura/farmacologia , Feminino , Técnicas de Transferência Nuclear , Partenogênese/efeitos dos fármacos , Fatores de TempoRESUMO
Pituitary adenomas usually develop temozolomide resistance, which could compromise the anticancer effects of temozolomide. Suppression of hypoxia-inducible factor 1α has been shown to sensitize glioblastoma cells to temozolomide treatment according to previous reports. However, whether and how the suppression of hypoxia-inducible factor 1α could sensitize pituitary adenomas to temozolomide treatment are still poorly understood. In the present study, using hypoxia-inducible factor 1α knockdown strategy, we demonstrated for the first time that hypoxia-inducible factor 1α knockdown could inhibit temozolomide-induced autophagy in rat pituitary adenoma GH3 cells and thus increase antitumor efficacy of temozolomide. Furthermore, we found hypoxia-inducible factor 1α knockdown could block autophagy process through neutralizing lysosomal pH value but not inhibiting autophagy induction. Finally, we found hypoxia-inducible factor 1α could regulate lysosomal pH value through regulating full length presenilin 1 expression, and exogenous reexpression of presenilin 1could restore lysosome acidic levels. Our data indicated hypoxia-inducible factor 1α knockdown could be a potential approach to improve the efficacy of temozolomide therapy for pituitary adenomas.