RESUMO
BACKGROUND: Multiple carpometacarpal fractures and dislocations are rare. This case report describes a novel multiple carpometacarpal injury, namely, 'diagonal' carpometacarpal joint fracture and dislocation. CASE PRESENTATION: A 39-year-old male general worker sustained a compression injury to his right hand in the dorsiflexion position. Radiography indicated a Bennett fracture, hamate fracture, and fracture at the base of the second metacarpal. Subsequent computed tomography and intraoperative examination confirmed an injury to the first to fourth carpometacarpal joint along a diagonal line. The normal anatomy of the patient's hand was successfully restored via open reduction combined with Kirschner wire and steel plate fixation. CONCLUSION: Our findings highlight the importance of taking the injury mechanism into account to avoid a missed diagnosis and to choose the best treatment approach. This is the first case of 'diagonal' carpometacarpal joint fracture and dislocation to be reported in the literature.
Assuntos
Articulações Carpometacarpais , Fraturas Ósseas , Fraturas Múltiplas , Traumatismos da Mão , Luxações Articulares , Traumatismo Múltiplo , Traumatismos do Punho , Masculino , Humanos , Adulto , Articulações Carpometacarpais/diagnóstico por imagem , Articulações Carpometacarpais/cirurgia , Fraturas Ósseas/complicações , Fraturas Ósseas/diagnóstico por imagem , Luxações Articulares/complicações , Luxações Articulares/diagnóstico por imagem , Traumatismos da Mão/cirurgiaRESUMO
OBJECTIVES: To explore the potential biomarkers for the diagnosis of primary brain stem injury (PBSI) by using metabonomics method to observe the changes of metabolites in rats with PBSI caused death. METHODS: PBSI, non-brain stem brain injury and decapitation rat models were established, and metabolic maps of brain stem were obtained by LC-MS metabonomics method and annotated to the HMDB database. Partial least square-discriminant analysis (PLS-DA) and random forest methods were used to screen potential biomarkers associated with PBSI diagnosis. RESULTS: Eighty-six potential metabolic markers associated with PBSI were screened by PLS-DA. They were modeled and predicted by random forest algorithm with an accuracy rate of 83.3%. The 818 metabolic markers annotated to HMDB database were used for random forest modeling and prediction, and the accuracy rate was 88.9%. According to the importance in the identification of cause of death, the most important metabolic markers that were significantly up-regulated in PBSI group were HMDB0038126 (genipinic acid, GA), HMDB0013272 (N-lauroylglycine), HMDB0005199 [(R)-salsolinol] and HMDB0013645 (N,N-dimethylsphingosine). CONCLUSIONS: GA, N-lauroylglycine, (R)-salsolinol and N,N-dimethylsphingosine are expected to be important metabolite indicators in the diagnosis of PBSI caused death, thus providing clues for forensic medicine practice.
Assuntos
Lesões Encefálicas , Metabolômica , Ratos , Animais , Metabolômica/métodos , Biomarcadores/metabolismo , Tronco Encefálico/metabolismoRESUMO
l-Phenylalanine is an important amino acid that is widely used in the production of food flavors and pharmaceuticals. Generally, l-phenylalanine production by engineered Escherichia coli requires a high rate of oxygen supply. However, the coexpression of Vitreoscilla hemoglobin gene (vgb), driven bya tac promoter, with the genes encoding 3-deoxy-d-arabinoheptulosonate-7-phosphate synthetase (aroF) and feedback-resistant chorismate mutase/prephenate dehydratase (pheAfbr ), led to increased productivity and decreased demand for aeration by E. coli CICC10245. Shake-flask studies showed that vgb-expressing strains displayed higher rates of oxygen uptake, and l-phenylalanine production under standard aeration conditions was increased. In the aerobic fermentation process, cell growth, l-phenylalanine production, and glucose consumption by the recombinant E. coli strain PAPV, which harbored aroF, pheAfbr , and tac-vgb genes, were increased compared to that in the strain harboring only aroF and pheAfbr (E. coli strain PAP), especially under oxygen-limited conditions. The vgb-expressing strain PAPV produced 21.9% more biomass and 16.6% more l-phenylalanine, while consuming only approximately 5% more glucose after 48 H of fermentation. This study demonstrates a method to enhance the l-phenylalanine production by E. coli using less intensive and thus more economical aeration conditions.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Fenilalanina/biossíntese , Hemoglobinas Truncadas/genética , Hemoglobinas Truncadas/metabolismo , Proteínas de Bactérias/biossíntese , Fermentação , Fenilalanina/química , Fenilalanina/genética , Regiões Promotoras Genéticas/genética , Hemoglobinas Truncadas/biossínteseRESUMO
BACKGROUND: Aberrant vascular smooth muscle cell (VSMC) proliferation and migration contribute to the development of vascular pathologies, such as atherosclerosis and post-angioplasty restenosis. The aim of this study was to determine whether miR-22-3p plays a role in regulating human artery vascular smooth muscle cell (HASMC) function and neointima formation. METHODS: Quantitative real-time PCR (qRT-PCR) and fluorescence in situ hybridization (FISH) were used to detect miR-22-3p expression in human arteries. Cell Counting Kit-8 (CCK-8) and EdU assays were performed to assess cell proliferation, and transwell and wound closure assays were performed to assess cell migration. Moreover, luciferase reporter assays were performed to identify the target genes of miR-22-3p. Finally, a rat carotid artery balloon-injury model was used to determine the role of miR-22-3p in neointima formation. RESULTS: MiR-22-3p expression was downregulated in arteriosclerosis obliterans (ASO) arteries compared with normal arteries, as well as in platelet-derived growth factor-BB (PDGF-BB)-stimulated HASMCs compared with control cells. MiR-22-3p overexpression had anti-proliferative and anti-migratory effects and dual-luciferase assay showed that high mobility group box-1 (HMGB1) is a direct target of miR-22-3p in HASMCs. Furthermore, miR-22-3p expression was negatively correlated with HMGB1 expression in ASO tissue specimens. Finally, LV-miR-22-3p-mediated miR-22-3p upregulation significantly suppressed neointimal hyperplasia specifically by reducing HMGB1 expression in vivo. CONCLUSIONS: Our results indicate that miR-22-3p is a key molecule in regulating HASMC proliferation and migration by targeting HMGB1 and that miR-22-3p and HMGB1 may be therapeutic targets in the treatment of human ASO.
Assuntos
Arteriosclerose Obliterante/patologia , Proteína HMGB1/metabolismo , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Animais , Antagomirs/metabolismo , Arteriosclerose Obliterante/metabolismo , Sequência de Bases , Becaplermina , Lesões das Artérias Carótidas/metabolismo , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/veterinária , Movimento Celular , Proliferação de Células , Células Cultivadas , Proteína HMGB1/antagonistas & inibidores , Proteína HMGB1/genética , Humanos , Masculino , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Músculo Liso Vascular/citologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Proteínas Proto-Oncogênicas c-sis/farmacologia , Ratos , Ratos Sprague-Dawley , Alinhamento de SequênciaRESUMO
BACKGROUND/AIMS: ALT1 is a novel long non-coding RNA derived from the alternatively spliced transcript of the deleted in lymphocytic leukemia 2 (DLEU2). To date, ALT1 biological roles in human vascular endothelial cells have not been reported. METHODS: ALT1 was knocked down by siRNAs. Cell proliferation was analyzed by cck-8. The existence and sequence of human ALT1 were identified by 3' rapid amplification of cDNA ends. The interaction between lncRNA and proteins was analyzed by RNA-Protein pull down assay, RNA immunoprecipitation, and mass spectrometry analysis. RESULTS: ALT1 was expressed in human umbilical vein endothelial cells (HUVECs). The expression of ALT1 was significantly downregulated in contact-inhibited HUVECs and in hypoxia-induced, growth-arrested HUVECs. Knocking down of ALT1 inhibited the proliferation of HUVECs by G0/G1 cell cycle arrest. We observed that angiotensin converting enzyme â ¡(ACE2) was a direct target gene of ALT1. Knocking-down of ALT1 or its target gene ACE2 could efficiently decrease the expression of cyclin D1 via the enhanced ubiquitination and degradation, in which HIF-1α and protein von Hippel-Lindau (pVHL) might be involved. CONCLUSION: The results suggested the human long non-coding RNA ALT1 is a novel regulator for cell cycle of HUVECs via ACE2 and cyclin D1 pathway.
Assuntos
Peptidil Dipeptidase A/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Enzima de Conversão de Angiotensina 2 , Apoptose , Proteínas de Transporte/metabolismo , Hipóxia Celular , Proliferação de Células , Proteínas Culina/antagonistas & inibidores , Proteínas Culina/genética , Proteínas Culina/metabolismo , Ciclina D1/genética , Ciclina D1/metabolismo , Proteínas do Citoesqueleto , Regulação para Baixo , Pontos de Checagem da Fase G1 do Ciclo Celular , Células Endoteliais da Veia Umbilical Humana , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Imunoprecipitação , MicroRNAs/metabolismo , Chaperonas Moleculares , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/genética , Interferência de RNA , RNA Longo não Codificante , RNA Interferente Pequeno/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Transferases , Proteínas Supressoras de Tumor/antagonistas & inibidores , Proteínas Supressoras de Tumor/genética , UbiquitinaçãoRESUMO
Fetal growth restriction (FGR) threatens perinatal health and is correlated with increased incidence of fetal original adult diseases. Most cases of FGR were idiopathic, which were supposed to be associated with placental abnormality. Decreased circulating placental growth factor (PGF) was recognized as an indication of placental deficiency in FGR. In this study, the epigenetic regulation of PGF in FGR placentas and the involvement of PGF in modulation of trophoblast activity were investigated. The expression level of PGF in placental tissues was determined by RT-qPCR, immunohistochemistry and ELISA. DNA methylation profile of PGF gene was analyzed by bisulfite sequencing. Trophoblastic cell lines were treated with ZM-306416, an inhibitor of PGF receptor FLT1, to observe the effect of PGF/FLT1 signaling on cell proliferation and migration. We demonstrated that PGF was downregulated in placentas from FGR pregnancies compared with normal controls. The villous expression of PGF was positively correlated with placental and fetal weight. The CpG island inside PGF promoter was hypomethylated without obvious difference in both normal and FGR placentas. However, the higher DNA methylation at another CpG island downstream exon 7 of PGF was demonstrated in FGR placentas. Additionally, we found FLT1 was expressed in trophoblast cells. Inhibition of PGF/FLT1 signaling by a selective inhibitor impaired trophoblast proliferation and migration. In conclusion, our data suggested that the PGF expression was dysregulated, and disrupted PGF/FLT1 signaling in trophoblast might contribute to placenta dysfunction in FGR. Thus, our results support the significant role of PGF in the pathogenesis of FGR.
Assuntos
Metilação de DNA , Retardo do Crescimento Fetal/etiologia , Regulação da Expressão Gênica no Desenvolvimento , Doenças Placentárias/fisiopatologia , Fator de Crescimento Placentário/metabolismo , Trofoblastos/metabolismo , Adulto , Peso ao Nascer , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ilhas de CpG/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Éxons/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Doenças Placentárias/sangue , Doenças Placentárias/metabolismo , Doenças Placentárias/patologia , Fator de Crescimento Placentário/antagonistas & inibidores , Fator de Crescimento Placentário/genética , Placentação , Gravidez , Regiões Promotoras Genéticas/efeitos dos fármacos , Quinazolinas/farmacologia , Interferência de RNA , Trofoblastos/efeitos dos fármacos , Trofoblastos/patologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
OBJECTIVE: The aims of this study were to make clear whether miR-21 was dysregulated in hydatidiform mole (HM) tissues and choriocarcinoma (CCA) cells, to elucidate whether aberrant miR-21 expression would affect the function of CCA cells, and to find out whether there was a relationship between miR-21 and AKT, PDCD4, and PTEN in CCA cells. METHODS: Fresh and formalin-fixed, paraffin-embedded trophoblastic tissues (normal first trimester placentas and HMs) were retrieved from the biobank in the International Peace Maternity and Child Health Hospital, Shanghai Jiao Tong University. Choriocarcinoma JAR and JEG-3 cells were cultured. Expression of miR-21 in trophoblast cells and tissues was examined by quantitative real-time polymerase chain reaction. Location and distribution of miR-21 in trophoblast tissues were determinated by in situ hybridization and fluorescent in situ hybridization. The effect of miR-21 on JAR and JEG-3 cells was tested by miR-21 mimics and inhibitor transfection, followed by cell viability assay, flow cytometric analysis, and Transwell analysis. Interaction between miR-21 and its target genes in CCA cells was verified by quantitative real-time polymerase chain reaction, Western blot, and luciferase report system. RESULTS: We originally found miR-21 was markedly upregulated in HM tissues compared with normal first trimester placentas. The expression of miR-21 was exclusively confined in trophoblastic layers. Furthermore, we discovered miR-21 was significantly increased in JAR and JEG-3 cells compared with normal primary human trophoblastic cells. Moreover, we demonstrated miR-21 could promote proliferation, migration, and invasion of CCA cells. We furthermore proved miR-21 negatively regulated PDCD4 and PTEN in CCA cells and targeted to PDCD4 3'UTR directly. In addition, we confirmed that miR-21 could activate Akt pathway by phosphorylating Akt at Ser 473. CONCLUSIONS: Our results suggested miR-21 was responsible for aggressive phenotype of gestational trophoblastic disease and had the potential diagnostic and therapeutic values for gestational trophoblastic neoplasm.
Assuntos
Coriocarcinoma/genética , Coriocarcinoma/patologia , Mola Hidatiforme/genética , Mola Hidatiforme/patologia , MicroRNAs/biossíntese , Neoplasias Uterinas/genética , Neoplasias Uterinas/patologia , Proteínas Reguladoras de Apoptose/genética , Materiais Biomiméticos/farmacologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Coriocarcinoma/metabolismo , Feminino , Humanos , Mola Hidatiforme/metabolismo , Hibridização In Situ , Hibridização in Situ Fluorescente , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Invasividade Neoplásica , PTEN Fosfo-Hidrolase/genética , Fosforilação , Gravidez , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas de Ligação a RNA/genética , Transfecção , Regulação para Cima , Neoplasias Uterinas/metabolismoRESUMO
The objective of present study was to find out an accurate, rapid and nondestructive method to detect total acid content (TA) of pitaya with visible/near-infrared spectrometry, wavelet transform (WT) and successive projections algorithm (SPA), which will provide scientific basis for non- destructive measurement of pitaya. Maya2000 fiber-optic spectrumeter was used to collect spectral data of pitaya on the wavelength in the range of 380~1 099 nm; and then with the methods of WT denosing pretreatment, SPA and partial least squares regression (PLSR) quantitative forecasting model of TA of pitaya was established. The result showed that the precision of WT-SPA-PLSR model, which combine the WT with SPA, was better than that of PLSR model based on the whole wave variables. The relation coefficient of the PLSR model (Rp) that predicted TA based on the original spectrum of all samples as the input variables was 0.851 394 and RMSEP was 0.086 848. The original spectrum variable of the all samples were processed by using wavelet function dbN(N=2, 3, , 10) for wavelet decomposition and de-noising. The optimal results of noise reduction were decomposed in level 2 using wavelet function db4 (db4-2). The Rp of WT-PLSR model was 0.915 635 and RMSEP was 0.066 752. The prediction of model using wavelet transform de-noising was improved significantly. After the original spectrum processed by db10-3 and SPA, 12 preferred variables were selected from 570 spectrum variables, such as 530, 545, 604, 626, 648, 676, 685, 695, 730, 897, 972, 1 016 nm spectrum variables. The WT-SPA-PLSR model based on these 12 variables as input variables was established. Rp of the WT-SPA-PLSR prediction model was 0.882 83 and RMSEP was 0.077 39. SPA algorithm was suitable for the selection of spectrum variables which could effectively obtain the spectrum variables which were strong correlation with TA and increase the accuracy and stability of the prediction model. The results indicated that the nondestructive detection for TA of pitaya based on the diffuse reflectance visible/near-infrared spectrometry, WT and SPA was feasible.
RESUMO
AIMS: To explore the expression of miR-24-3p in human arteries with arteriosclerosis obliterans (ASO) as well as the role of miR-24-3p in the pathogenesis of ASO. METHODS: We used quantitative real-time PCR (qRT-PCR) and in situ hybridization to monitor miR-24-3p expression in human arteries. To investigate the effect of miR-24-3p on human arterial smooth muscle cells (HASMCs), we applied cell counting and EdU assays to monitor proliferation and transwell and wound healing assays to investigate migration and flow cytometry to investigate apoptosis. Furthermore, we applied 3'-untranslated region (3'-UTR) luciferase assays to investigate the role of miR-24-3p in targeting platelet-derived growth factor receptor B (PDGFRB) and c-Myc. RESULTS: MiR-24-3p was mainly located in the media of arteries and was downregulated in ASO arteries compared with normal arteries. Platelet-derived growth factor BB (PDGF-BB) treatment reduced the expression of miR-24-3p in primary cultured HASMCs. MiR-24-3p mimic oligos inhibited the proliferation and migration, and promotes apoptosis of HASMCs. Our 3'-UTR luciferase assays confirmed that PDGFRB and c-Myc were targets of miR-24-3p. CONCLUSION: The results suggest that miR-24-3p regulates the proliferation and migration of HASMCs by targeting PDGFRB and c-Myc. The PDGF/miR-24-3p/PDGFRB and PDGF/miR-24-3p/c-Myc pathways may play critical roles in the pathogenesis of ASO. These findings highlight the potential for new therapeutic targets for ASO.
Assuntos
Apoptose , Arteriosclerose Obliterante/genética , Movimento Celular , Proliferação de Células , Regulação da Expressão Gênica , MicroRNAs/genética , Miócitos de Músculo Liso/patologia , Arteriosclerose Obliterante/patologia , Sequência de Bases , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miócitos de Músculo Liso/metabolismo , Proteínas Proto-Oncogênicas c-myc/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genéticaRESUMO
BACKGROUND: Anatomical sublobar resection was widely performed for small-sized nodule located in the deep field of the lung. In this study, we compared surgical outcomes between anatomical sublobar resection in left upper lobe including segmentectomy and subsegmentectomy. We also applied a technique based on bronchovascular patterns for subsegmentectomy. PATIENTS AND METHODS: A hundred and fifty-one patients underwent anatomical sublobar resection of left upper lobe in our hospital in the period from January 2018 to December 2019 and they were retrospectively reviewed. Patients' characteristics and surgical outcome of the subsegmentectomy group (n = 71) were analyzed and compared to those of patients of the segmentectomy group (n = 80). The bronchovascular patterns of left upper lobe in these patients were also classified, and a technique of subsegmentectomy was introduced by cases description. RESULTS: Compared to segmentectomy, the subsegmentectomy group had longer operative time [141min, interquartile range (IQR): 125-161 vs. 127.5 min, interquartile range (IQR): 114.75-148.25; P = 0.001] and more estimated blood loss [50 mL, IQR: 30-50 vs. 30 mL, IQR: 20-50; P = 0.02]. Branching pattern of bronchus was classified into 3 types in left upper division, and 2 types in lingular division. Branching pattern of pulmonary artery was classified into 7 types in left upper division and 2 types in lingular division. Branching pattern of pulmonary vein was classified into 3 types in left upper division and V3 b was classified into 4 types. CONCLUSION: Subsegmentectomy is a safe procedure with surgical outcome comparable to the one of segmentectomy except operative time and estimated blood loss. Subsegmentectomy based on bronchovascular patterns in left upper lobe is feasible.
Assuntos
Neoplasias Pulmonares , Pneumonectomia , Humanos , Pneumonectomia/métodos , Neoplasias Pulmonares/cirurgia , Estudos Retrospectivos , Pulmão/irrigação sanguínea , Resultado do TratamentoRESUMO
The rapid and convenient method for identification of all kinds of G-quadruplex is highly desirable. In the present study, a novel colorimetric indicator for a vast variety of G-quadruplex was designed and synthesized on the basis of thiazole orange and isaindigotone skeleton. Its distinct color change enables label-free visual detection of G-quadruplexes, which is due to the disassembly of dye H-aggregates to monomers. This specific detection of G-quadruplex arises from its end-stacking interaction with G-quartet. On the basis of this universal indicator, a facile approach for large-scale identification of G-quadruplex was developed.
Assuntos
Alcaloides/química , Benzotiazóis/química , Colorimetria , Quadruplex G , Quinazolinas/química , Quinolinas/química , Transferência Ressonante de Energia de Fluorescência , Indicadores e Reagentes/químicaRESUMO
INTRODUCTION: Gestational trophoblastic disease (GTD) encompasses a range of trophoblastic disorders from hydatidiform mole (HM), to malignant gestational trophoblastic neoplasia (GTN). The exact molecular mechanisms of GTN remain unknown. Dysregulation and dysfunction of programmed cell death 4 (PDCD4)have been observed in many cancers. The roles of PDCD4 in GTD have not been previously reported. METHODS: A total of 161 cases of formalin-fixed, paraffin-embedded trophoblast blocks, and 36 cases of fresh trophoblast tissues were collected, including normal first trimester placentas, HM, and invasive HM. Choriocarcinoma cells JAR and JEG-3 were employed. The expressions of PDCD4 and small ubiquitin-like modifier 2/3 (SUMO2/3) were examined by immunohistochemistry, quantitative reverse transcription PCR and Western blotting in trophoblastic tissues and cells. The relationship between SUMOylation and PDCD4 was investigated. The effects of PDCD4 on proliferation, invasion, and migration of choriocarcinoma cells were evaluated by Cell Counting Kit-8 and transwell assays post siRNA transfection. Extracellular Matrix & Adhesion Profiler PCR Array was used to screen the downstream molecules of PDCD4. RESULTS: PDCD4 was significantly repressed in HM tissues. Loss of PDCD4 expression was demonstrated in 90% invasive HMs. Choriocarcinoma cells also displayed with suppressed PDCD4 expression. The varied expression of PDCD4 was paralleled by SUMO2/3. Inhibition of SUMOylation reduced the expression of PDCD4. Silencing of PDCD4promoted proliferation/migration/invasion, upregulatedMMP3/MMP8/ITGB2, and downregulated TIMP1/TIMP2 in choriocarcinoma cells. DISCUSSION: Our results suggest that reduced SUMOylation is one reason for suppressed PDCD4 in GTD. Loss of PDCD4 likely determines the malignant phenotype of GTN by dysregulating some members of the MMPs/TIMPs/integrins complex.
Assuntos
Coriocarcinoma , Doença Trofoblástica Gestacional , Mola Hidatiforme , Neoplasias Uterinas , Feminino , Humanos , Gravidez , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Coriocarcinoma/patologia , Regulação para Baixo , Doença Trofoblástica Gestacional/patologia , Mola Hidatiforme/patologia , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Neoplasias Uterinas/patologiaRESUMO
A series of 2-phenyl-benzopyranopyrimidine (PBPP) derivatives with alkylamino side chains were synthesized and found to be a new type of highly selective ligand to bind with telomeric G-quadruplex DNA, and their biological properties were reported for the first time. Their interactions with telomeric G-quadruplex DNA were studied with FRET melting, surface plasmon resonance, CD spectroscopy, and molecular modeling. Our results showed that the disubstituted PBPP derivatives could strongly bind to and effectively stabilize the telomeric G-quadruplex structure, and had significant selectivity for G-quadruplex over duplex DNA. In comparison, the mono substituted derivatives had much less effect on the G-quadruplex, suggesting that the disubstitution of PBPP is essential for its interaction with the G-quadruplex. Furthermore, telomerase inhibition of the PBPP derivatives and their cellular effects were studied, and compound 11b was found to be the most promising compound as a telomerase inhibitor and telomeric G-quadruplex binding ligand for further development for cancer treatment.
Assuntos
Benzopiranos/química , Quadruplex G , Pirimidinas/química , Telômero/química , Benzopiranos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Ligantes , Modelos Moleculares , Pirimidinas/farmacologia , Telomerase/antagonistas & inibidoresRESUMO
Deacetoxy/deacetylcephalosporin C synthase (acDAOC/DACS) from Acremonium chrysogenum is a bifunctional enzyme that catalyzes both the ring-expansion of penicillin N to deacetoxycephalosporin C and the hydroxylation of the latter to deacetylcephalosporin C. The R308 residue located in close proximity to the C-terminus of acDAOC/DACS was mutated to the other 19 amino acids. In the resulting mutant pool, R308L, R308I, R308T and R308V showed significant improvement in their ability to convert penicillin analogs, thus confirming the role of R308 in controlling substrate selectivity, the four amino acids all possess short aliphatic sidechains that may improve hydrophobic interactions with the substrates. The mutant R308I showed the highest reactivity for penicillin G, with 3-fold increase in k(cat)/K(m) ratio and 7-fold increase in relative activity.
Assuntos
Acremonium/enzimologia , Acremonium/genética , Oxirredutases/genética , Cromatografia Líquida de Alta Pressão , Mutagênese , Mutação , Penicilina G/análogos & derivados , Penicilina G/química , Especificidade por Substrato/genéticaRESUMO
BACKGROUND: Primary small cell esophageal carcinoma (PSCEC) is aggressive and rare, with a worse prognosis than other subtypes esophageal carcinoma. No definitive and optimum standard guidelines are established for treating it. Herein, we report a case of PSCEC, including a current literature review of PSCEC. CASE SUMMARY: A 79-year-old male was diagnosed PSCEC with multiple lymph node metastasis thorough computed tomography, positron emission tomography-computed tomography, endoscopy and pathology. Surgery was not suitable for this patient. He was treated with etoposide 100 mg/m2 and cisplatin 25 mg/m2 on days 1-3, every 3 wk for 4 cycles. The tumor and lymph nodes became smaller and dysphagia and vomiting symptoms improved. The patient could not tolerate subsequent chemotherapy (CT) because of hematological toxicity; therefore, we performed immunotherapy (durvalumab, 1500 mg) every 4 wk. At present the patient has received 12 cycles immunotherapy over about 1 year. He is still receiving treatment and follow-up. CONCLUSION: PSCEC with multiple lymph nodes metastasis does not always indicate surgery. CT may extend survival time and improve the quality of life in the absence of surgery. Immunotherapy or immunotherapy plus CT may also work as a treatment for PSCEC.
RESUMO
INTRODUCTION: In this study we examined the hypothesis that a hypoxic intrauterine environment causes mitochondrial dysfunction of trophoblasts in fetal growth restriction (FGR). METHODS: The mtDNA content, mRNA levels of mitochondrial encoded genes (ND6, COX I), mitochondrial membrane proteins (COX I, COX IV and VDAC), HIF-1α and BINP3 (mitophagy receptor) protein levels were examined in FGR placentas and normal placentas. The mitochondrial function (ATP production and mitochondrial membrane potential-ΔΨm) and above related proteins were further examined in hypoxic HTR-8/SVneo cells induced by cobalt chloride (CoCl2). Mitophagy and its regulating mechanism under hypoxia in FGR was also investigated. RESULTS: Compared with normal controls, both FGR placentas and CoCl2-treated trophoblast cells demonstrated statistically lower mtDNA content, reduced mRNAs of mitochondrial encoding genes, and decreased mitochondrial membrane proteins, accompanied by increased HIF-1α. Mitochondrial functions were impaired as demonstrated by decreased ATP production, and, reduced ΔΨm in CoCl2-treated cells. Meanwhile, mitophagy was markedly enhanced as indicated by increased LC3 fluorescent puncta in mitochondria of hypoxic trophoblastic cells. The upregulated BINP3 expression was demonstrated in FGR placentas as well as in hypoxic trophoblastic cells. DISCUSSION: We demonstrated that hypoxic conditions lead to impaired mitochondrial function in trophoblasts in FGR. Reduced mtDNA may be associated with enhanced mitophagy via activating HIF-1α/BINP3 signalling pathway, that may, in turn, affect nutrition and energy transfer to the growth-restricted fetus.
Assuntos
Retardo do Crescimento Fetal/metabolismo , Hipóxia/metabolismo , Mitocôndrias/metabolismo , Placenta/metabolismo , Adulto , Feminino , Humanos , Gravidez , Trofoblastos/metabolismoRESUMO
BACKGROUND: Dedifferentiated liposarcoma in the mediastinum is an extremely rare malignant neoplasm. A few previous case reports indicate that surgical resection is the major treatment, but frequent recurrence occurs locally. Due to its rarity, its clinical characteristics, optimal treatment and clinical outcomes remain unclear. Here, we report a case of multifocal recurrent dedifferentiated liposarcoma in the posterior mediastinum treated by combining surgery with 125I brachytherapy, and summarize its clinical features, treatment and prognosis. CASE SUMMARY: A 75-year-old man was admitted to our hospital with a history of gradual dysphagia for one year and aggravated dysphagia for 3 mo. Contrast-enhanced computed tomography (CT) revealed several large cystic-solid masses with lipomatous density, and calcification in the posterior-inferior mediastinum. The patient received a wide excision by video-assisted thoracoscopic surgery. Pathological analysis confirmed the tumors were dedifferentiated liposarcomas. The tumor locally relapsed 24 mo later, and another operation was performed by video-assisted thoracoscopic surgery. Fifteen months after the second surgery, the tumor recurred again, and the patient received CT-guided radioactive seeds 125I implantation. After 8 mo, follow-up chest CT showed an enlarged tumor. Finally, his condition exacerbated with severe dysphagia and dyspnea, and he died of respiratory failure in July 2018. CONCLUSION: We reviewed the literature, and suggest that surgical resection provides beneficial effects for dedifferentiated liposarcoma in the mediastinum, even in cases with local recurrence. 125I brachytherapy may be beneficial for recurrent unresectable patients.
RESUMO
BACKGROUND: Non-intubated video-assisted thoracoscopic surgery (NIVATS) has been increasingly used in lobectomy, bullectomy, wedge resection, lung volume reduction, sympathectomy and talc pleurodesis, which may reduce postoperative complications. However, the benefits of non-intubated and intubated methods of VATS remain controversial. METHODS: We comprehensively searched PubMed, Web of Science, Embase and the Cochrane Library, and performed a systematic review to assess the two techniques. Random and fixed-effects meta-analytical models were used based on the low between-study heterogeneity. Study quality, publication bias, and heterogeneity were assessed. RESULTS: Compared to intubated methods, NIVATS had a lower postoperative complications rate [odds ratio (OR): 0.63; 95% confidence interval (CI), 0.46-0.86; P<0.01], shorter global in-operating time [weighted mean difference (WMD): -35.96 min; 95% CI, -48.00 to -23.91; P<0.01], shorter hospital stay (WMD: -1.35 days; 95% CI, -1.72 to -0.98; P<0.01), shorter anesthesia time (WMD: -7.29 min; 95% CI, -13.30 to -1.29; P<0.01), shorter chest-tube placement time (WMD: -1.04 days; 95% CI, -1.75 to -0.33; P<0.01), less chest pain (WMD: -1.31; 95% CI, -2.45 to -0.17; P<0.05) and lower perioperative mortality rate (OR: 0.13; 95% CI, 0.02-0.99; P=0.05). CONCLUSIONS: NIVATS is a safe, efficient and feasible technique for thoracic surgery and may be a better alternative procedure owing to its advantage in reducing postoperative complications rate, hospital stay, and chest pain.
RESUMO
Gestational trophoblastic disease (GTD) encompasses a range of trophoblast-derived disorders. The most common type of GTD is hydatidiform mole (HM). Some of HMs can further develop into malignant gestational trophoblastic neoplasia (GTN). Aberrant expression of microRNA (miRNA) is widely reported to be involved in the initiation and progression of cancers. MiRNA expression profile also has been proved to be the useful signature for diagnosis, staging, prognosis, and response to chemotherapy. Till now, the profile of miRNA in the progression of GTD has not been determined. In this study, a total of 34 GTN and 60 complete HMs (CHM) trophoblastic tissues were collected. By miRNA array screening and qRT-PCR validating, six miRNAs, including miR-370-3p, -371a-5p, -518a-3p, -519d-3p, -520a-3p, and -934, were identified to be differentially expressed in GTN vs. CHM. Functional analyses further proved that miR-371a-5p and miR-518a-3p promoted proliferation, migration, and invasion of choriocarcinoma cells. Moreover, we demonstrated that miR-371a-5p was negatively related to protein levels of its predictive target genes BCCIP, SOX2, and BNIP3L, while miR-518a-3p was negatively related to MST1 and EFNA4. For the first time, we proved that miR-371a-5p and miR-518a-3p directly targeted to 3'-UTR regions of BCCIP and MST1, respectively. Additionally, we found that miR-371a-5p and miR-518a-3p regulated diverse pathways related to tumorigenesis and metastasis in choriocarcinoma cells. The results presented here may offer new clues to the progression of GTD and may provide diagnostic biomarkers for GTN.