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Hydrogen sulfide (H2S) is a gaseous signaling molecule reported to play multiple roles in fruit ripening. However, the molecular mechanisms underlying H2S-mediated delay in fruit ripening remain to be established. Here, the gene encoding a WRKY transcription factor, WRKY71, was identified as substantially upregulated in H2S-treated tomato (Solanum lycopersicum) via transcriptome profiling. The expression of WRKY71 was negatively associated with that of CYANOALANINE SYNTHASE1 (CAS1). Transient and stable genetic modification experiments disclosed that WRKY71 acts as a repressor of the tomato ripening process. CAS1 appears to play an opposite role, based on the finding that the ripening process was delayed in the cas1 mutant and accelerated in CAS1-OE tomatoes. Dual-luciferase reporter assay, yeast one-hybrid, electrophoretic mobility shift assay, and transient transformation experiments showed that WRKY71 bound to the CAS1 promoter and suppressed its activation. Moreover, the persulfidation of WRKY71 enhanced its binding ability to the CAS1 promoter. Data from luciferase complementation and Y2H assays confirmed that WRKY71 interacts with a BOI-related E3 ubiquitin-protein ligase 3 (BRG3) and is ubiquitinated in vitro. Further experiments showed that modification of BRG3 via persulfidation at Cys206 and Cys212 led to reduced ubiquitination activity. Our findings support a model whereby BRG3 undergoes persulfidation at Cys206 and Cys212, leading to reduced ubiquitination activity and decreased interactions with the WRKY71 transcript, with a subsequent increase in binding activity of the persulfidated WRKY71 to the CAS1 promoter, resulting in its transcriptional inhibition and thereby delayed ripening of tomatoes. Our collective findings provide insights into a mechanism of H2S-mediated regulation of tomato fruit ripening.
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Solanum lycopersicum , Solanum lycopersicum/genética , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação , Regiões Promotoras Genéticas/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Frutas/genética , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Etilenos/metabolismoRESUMO
PURPOSE: The aim of the present study was to compare 45° and 60° of sagittal femoral tunnel angles in terms of anterior tibial translation (ATT), valgus angle and graft in situ force following anterior cruciate ligament reconstruction (ACLR). METHODS: Ten porcine knees were subjected to the following loading conditions: (1) 89 N anterior tibial load at 35° (full extension), 60° and 90° of knee flexion and (2) 5 N m valgus tibial moment at 35° and 45° of knee flexion. ATT and graft in situ force of the intact anterior cruciate ligament (ACL) and ACLR were collected using a robotic universal force/moment sensor (UFS) testing system for (1) ACL intact, (2) ACL-deficient (ACLD) and (3) two different ACLR using different sagittal femoral tunnel angles (coronal 45°/sagittal 45° and coronal 45°/sagittal 60°). RESULTS: During the anterior tibial load, the femoral tunnel angle of ACLR knees at coronal 45°/sagittal 45° and 60° had significantly higher ATT than that of the ACL-intact knees at 60° of knee flexion (p < 0.05). The femoral tunnel angle of ACLR knees at coronal 45°/sagittal 60° had significantly lower graft in situ force than that of the ACL-intact knees at 60° and 90° of knee flexion (p < 0.05). During the valgus tibial moment, the femoral tunnel angle of ACLR knees at coronal 45°/sagittal 45° and 60° had significantly lower graft in situ force than that of the ACL-intact knees at all knee flexions (p < 0.05). CONCLUSIONS: The femoral tunnel angle of ACLR knees at coronal 45°/sagittal 45° provided similar ATT, valgus angle and graft in situ force to that of ACLR knees at coronal 45°/sagittal 60°. Therefore, both femoral tunnel angles could be used in ACLR, as the sagittal femoral tunnel angle does not appear to be relevant in post-operative knee stability. LEVEL OF EVIDENCE: Not applicable.
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Ethylene is a key phytohormone that regulates the ripening of climacteric fruits, and methionine is an indirect precursor of ethylene. However, whether methionine synthase plays a role in fruit ripening in Solanum lycopersicum (tomato) is still unknown. In this study, we find that a tomato methionine synthase (named SlMS1), which could be repressed at the transcriptional level by hydrogen sulfide (H2S), acts as a positive regulator for tomato fruit ripening. By a bioinformatics analysis, it is found that SlMS1 and SlMS2 in tomato are highly homologous to methionine synthases in Arabidopsis thaliana. The expression pattern of SlMS1 and SlMS2 is analyzed in tomato, and SlMS1 expression is up-regulated during fruit ripening, suggesting its potential role in regulating fruit ripening. A potential bipartite nuclear localization signal is found in the amino acid sequence of SlMS1; thus, SlMS1 is tagged with GFP and observed in the leaves of Nicotiana benthamiana. Consistently, SlMS1-GFP shows strong nuclear localization and also cytoplasmic localization. The role of SlMS1 in regulating fruit ripening is investigated in tomato fruit by transient silencing (virus-induced gene silencing, VIGS) and transient overexpression. The results show that SlMS1 silencing causes delayed fruit ripening, evidenced by more chlorophyll and less carotenoid accumulation, while SlMS1 overexpression accelerates fruit ripening significantly compared with control. Further investigation shows that SlMS1 overexpression could up-regulate the expression of carotenoid-synthesis-related genes (PSY1, PDS, ZDS), chlorophyll-degradation-related genes (NYC1, PAO, PPH, SGR1), cell-wall-metabolism-related genes (CEL2, EXP, PG, TBG4, XTH5) and ethylene-synthesis-pathway-related genes (ACO1, ACO3, ACS2), while SlMS1 silencing causes the opposite results. The correlation analysis indicates that SlMS1 expression is negatively correlated with chlorophyll content and positively correlated with carotenoid and ripening-related gene expressions. Taken together, our data suggest that SlMS1 is a positive regulator of tomato fruit ripening and a possible target gene for the ripening-delaying effect of H2S.
Assuntos
Sulfeto de Hidrogênio , Solanum lycopersicum , Solanum lycopersicum/metabolismo , Frutas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Sulfeto de Hidrogênio/metabolismo , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , Sinais de Localização Nuclear/genética , Proteínas de Plantas/metabolismo , Etilenos/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Metionina/metabolismo , Hidrogênio/metabolismo , Sulfetos/metabolismoRESUMO
Calcium deficiency usually causes accelerated quality deterioration in postharvest fruit, whereas the underlining mechanism is still unclear. Here, we report that calcium deficiency induced the development of bitter pit on the surface of apple peels compared with the healthy appearance in control apples during postharvest storage. Physiological analysis indicates that calcium-deficient peels contained higher levels of superoxide anion (O2â¢-), malondialdehyde (MDA), total phenol, flavonoid contents and polyphenol oxidase (PPO) activity, and reduced calcium, H2S production, anthocyanin, soluble protein content, and peroxidase (POD) activity compared with those in calcium-sufficient peels. The principal component analysis (PCA) results show that calcium content, ROS, and H2S production were the main factors between calcium-deficient and calcium-sufficient apple peels. Transcriptome data indicated that four calmodulin-like proteins (CMLs), seven AP2/ERFs, and three bHLHs transcripts were significantly differentially expressed in calcium-deficient apple peels. RT-qPCR and correlation analyses further revealed that CML5 expression was significantly positively correlated with the expression of ERF2/17, bHLH2, and H2S production related genes. In addition, transcriptional co-activation of CML5 by ERF2 and bHLH2 was demonstrated by apple transient expression assays and dual-luciferase reporter system experiments. Therefore, these findings provide a basis for studying the molecular mechanism of postharvest quality decline in calcium-deficient apples and the potential interaction between Ca2+ and endogenous H2S.
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Sulfeto de Hidrogênio/metabolismo , Malus/metabolismo , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transcriptoma , Antocianinas/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Cálcio/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Catecol Oxidase/metabolismo , Flavonoides/metabolismo , Armazenamento de Alimentos , Frutas/genética , Frutas/metabolismo , Malus/genética , Fatores de Terminação de Peptídeos/genética , Fatores de Terminação de Peptídeos/metabolismo , Fenóis/metabolismo , Fenótipo , Proteínas de Plantas/genética , Análise de Componente PrincipalRESUMO
BACKGROUND: Anthocyanins, which have important biological functions and have a beneficial effect on human health, notably account for pigmentation in purple-fleshed sweet potato tuberous roots. Individual regulatory factors of anthocyanin biosynthesis have been identified; however, the regulatory network of anthocyanin biosynthesis in purple-fleshed sweet potato is unclear. RESULTS: We functionally determined that IbMYB340 cotransformed with IbbHLH2 in tobacco and strawberry receptacles induced anthocyanin accumulation, and the addition of IbNAC56a or IbNAC56b caused increased pigmentation. Furthermore, we confirmed the interaction of IbMYB340 with IbbHLH2 and IbNAC56a or IbNAC56b via yeast two-hybrid and firefly luciferase complementation assays; these proteins could form a MYB340-bHLH2-NAC56a or MYB340-bHLH2-NAC56b transcriptional complex to regulate anthocyanin biosynthesis by binding to the IbANS promoter rather than the IbUFGT promoter. Furthermore, it was found by a transient expression system in tobacco leaves that IbMYB44 could decrease anthocyanin accumulation. Moreover, the interaction of IbMYB44 with IbMYB340 and IbNAC56a or IbNAC56b was verified. This result suggested that IbMYB44 acts as a repressor of anthocyanin in sweet potato. CONCLUSIONS: The repressor IbMYB44 affected anthocyanin biosynthesis by competitively inhibiting the IbMYB340-IbbHLH2-IbNAC56a or IbMYB340-IbbHLH2-IbNAC56b regulatory complex formation. Overall, the present study proposed a novel regulatory network whereby several vital TFs play key roles in regulating anthocyanin biosynthesis, and it provides strong insight into the potential mechanism underlying anthocyanin biosynthesis in sweet potato tuberous roots with purple color.
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Antocianinas/biossíntese , Ipomoea batatas/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Fragaria , Regulação da Expressão Gênica de Plantas/genética , Ipomoea batatas/genética , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Plantas Geneticamente Modificadas , Alinhamento de Sequência , Nicotiana , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologiaRESUMO
BACKGROUND: Sweet potato is susceptible to chilling injury during low-temperature storage. To explore the correlation between chilling injury and reactive oxygen species (ROS) metabolism, the content of ROS and the activities and gene expression of antioxidant enzymes were analyzed in the typical storage-tolerant cultivar Xushu 32 and storage-sensitive cultivar Yanshu 25. RESULTS: The activities of antioxidant enzymes including ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were enhanced rapidly in the early period of storage in response to chilling stress. Thereafter, the content of ROS metabolites increased consistently due to gradual decrease in ROS scavenging enzymes. Storage-tolerant cultivar Xushu 32 had higher antioxidant enzyme activities and gene expressions as well as higher content of antioxidant metabolites and lower content of ROS metabolites compared with storage-sensitive cultivar Yanshu 25, suggesting that the capacity of ROS scavenging by antioxidant enzymes and antioxidants is highly associated with the tolerance of sweet potato to chilling stress. CONCLUSION: These results indicated that the antioxidative system is activated in the storage root of sweet potato and the antioxidative capacity is positively associated with better storage performance in the storage-tolerant cultivar. © 2019 Society of Chemical Industry.
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Antioxidantes/metabolismo , Ipomoea batatas/enzimologia , Tubérculos/química , Ascorbato Peroxidases/genética , Ascorbato Peroxidases/metabolismo , Catalase/metabolismo , Temperatura Baixa , Armazenamento de Alimentos , Regulação da Expressão Gênica de Plantas , Glutationa Redutase/genética , Glutationa Redutase/metabolismo , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismoRESUMO
KEY MESSAGE: This research reveals the different expression patterns of anthocyanin biosynthesis enzyme genes and transcription factors in six red-skinned pear cultivars with different genetic backgrounds. Skin color is an important feature of pear fruits, with red skin generally attracting consumers. However, great differences of coloration exist in different pear cultivars. To elucidate the characteristics of pigmentation in pear cultivars with different genetic backgrounds, six cultivars, belonging to P. communis, P. pyrifolia, P. ussuriensis, P. bretschneideri, and a hybrid of P. communis × P. pyrifolia, were used to detect pigment concentrations, expressions of seven anthocyanin biosynthesis enzyme genes, and three related transcription factor genes. Results showed that the occidental pears 'Starkrimson' and 'Red Bartlett' colored at the beginning of fruit setting, but color decreased with fruit maturity; the other four cultivars showed low anthocyanin accumulations and the contents increased during fruit development, but also decreased at later stages. The expression patterns of genes encoding enzymes indicated that ANS and UFGT were decisive genes for anthocyanin biosynthesis for red-skinned pear, and their different expressions led to the coloration differences between occidental and oriental pears. The expression patterns of transcription factors indicated that the different co-expression of MYB10 and bHLH33 genes and the different expressions of WD40 are involved in the differential regulation mechanisms of anthocyanin biosynthesis and coloration pattern between occidental and oriental pears.
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Antocianinas/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Pyrus/genética , Frutas/genética , Frutas/metabolismo , Fenótipo , Pigmentação , Proteínas de Plantas/metabolismo , Pyrus/metabolismo , Especificidade da Espécie , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
OBJECTIVE: To describe the characteristics of acute pancreatitis during pregnancy and postpartum. METHODS: From 1994 to 2012, 18 cases of gravida and postpartum women complicated with acute pancreatitis were treated at Peking University Third Hospital and retrospective analysis was performed. RESULTS: The admission rate due to acute pancreatitis was 41.24 per 100 000 admission. The median age was 32.0 (28.0, 34.7) years, with the onset of acute pancreatitis on 35.0 (23.5, 37.0) weeks'gestation. Of all the cases, 13 (72.2%) were interstitial edematous acute pancreatitis, and 5 (27.8%) were necrotizing. Nine (50.0%) were caused by hyperlipidemia, of which 7 (38.9%) were caused by gallstones, and 2 (1.1%) were idiopathic. Stratified by severity, 6 (33.3%) were mild, 7 (38.9%) were moderately severe and 5 (27.8%) were severe, of which 8 were transferred to Intensive Care Unit. Compared with non-hyperlipidemic acute pancreatitis, cases caused by hyperlipidemia were more severe, however, the prognoses of mothers and infants were good. CONCLUSION: The incidence of acute pancreatitis during pregnancy and postpartum was similar to that of the regular population, and mostly caused by gallstones and hyperlipidemia. Although acute hyperlipidemic pancreatitis is more severe, the morbidity and mortality of maternal and perinatal could be decreased by aggressive therapy.
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Doença Aguda , Hiperlipidemias/complicações , Pancreatite/complicações , Período Pós-Parto , Complicações na Gravidez , Colelitíase , Feminino , Humanos , Incidência , Gravidez , Prognóstico , Estudos RetrospectivosRESUMO
Introduction: The biomechanical indication for combining anterolateral structures reconstruction (ASLR) with ACL reconstruction (ACLR) to reduce pivot shift in the knee remains unclear. This study aims to investigate knee functionality after ACL rupture with different combinations of injuries, and to compare the effectiveness of ALSR with ACLR for treating these injuries. Methods: A validated finite element model of a human cadaveric knee was used to simulate pivot shift tests on the joint in different states, including 1) an intact knee; 2) after isolated ACL rupture; 3) after ACL rupture combined with different knee injuries or defect, including a posterior tibial slope (PTS) of 20°, an injury to the anterolateral structures (ALS) and an injury to the posterior meniscotibial ligament of the lateral meniscus (LP); 4) after treating the different injuries using isolated ACLR; v. after treating the different injuries using ACLR with ALSR. The knee kinematics, maximum von Mises stress (Max.S) on the tibial articular cartilage (TC) and force in the ACL graft were compared among the different simulation groups. Results and discussion: Comparing with isolated ACL rupture, combined injury to the ALS caused the largest knee laxity, when a combined PTS of 20° induced the largest Max.S on the TC. The joint stability and Max.S on the TC in the knee with an isolated ACL rupture or a combined rupture of ACL and LP were restored to the intact level after being treated with isolated ACLR. The knee biomechanics after a combined rupture of ACL and ALS were restored to the intact level only when being treated with a combination of ACLR and ALSR using a large graft diameter (6 mm) for ALSR. However, for the knee after ACL rupture combined with a PTS of 20°, the ATT and Max.S on the TC were still greater than the intact knee even after being treated with a combination of ACLR and ALSR. The finite element analysis showed that ACLR should include ALSR when treating ACL ruptures accompanied by ALS rupture. However, pivot shift in knees with a PTS of 20° was not eliminated even after a combined ACLR and ALSR.
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OBJECTIVE: Isolated intra-articular anterior cruciate ligament (ACL) reconstruction is not capable of restoring instability in many cases leading some to recommend concomitant anterolateral ligament (ALL) reconstruction. The satisfactory fixation site and graft length change are crucial in ligament reconstruction to restore the ALL function and avoid some unwanted graft behavior. The purpose of this investigation is to determine the optimal isometric region on the anterolateral aspect of the knee for ALL reconstruction using a three-dimensional optical instrument and a suture similar to an intraoperative isometric test. METHODS: Six freshly frozen cadaveric human knees were used in this study. Data regarding the anterolateral surface were obtained using an optical measurement system to create a three-dimensional model. Nine points were selected on the femur (F1-F9) and tibia (Ta-Ti) respectively. The three-dimensional length change between each pair of tibial and femoral points was measured during passive knee flexion from 0° to 90° in 15° increments. Subsequently, five femoral points (A-E) were selected from the lateral femur, located in different areas relative to the lateral femoral epicondyle, and three tibial reference points (T1-T3) were selected in the isometric test. The changes in the length between each pair of reference points were measured using sutures. The 95% confidence interval for the rate of length change was estimated using the mean and standard deviation of the maximum rate of length change at different flexion angles, and the data were expressed as the mean (95% confidence interval) and compared with the maximum acceptable rate of change (10%). RESULTS: The maximum acceptable change rate for ligament reconstruction is 10%, and the mean maximum rates and the 95% confidence interval (CI) of length change for the point combinations were calculated. Among all the combined points measured using the optical measurement system and the suture, the qualified point combination for reconstruction was F3 (8mm posterior and 8mm proximal to the lateral femoral epicondyle)-Tb (8mm proximal to the midpoint between the center of Gerdy's tubercle and the fibula head), A (posterior and proximal to the lateral femoral epicondyle)-T2 (10mm below the joint line)and A-T3 (15 mm below the joint line). The position of F3-Tb and A-T2 are close to each other. CONCLUSION: The most isometric area of the femur for ALL reconstruction was posterior and proximal to the lateral femoral epicondyle. We recommend that the initial location of the femoral point be set at 8 mm posterior and 8 mm proximal to the lateral femoral epicondyle and the tibial point at approximately 10 mm below the joint line, midway between Gerdy's tubercle and fibular head, and subsequently adjusted to the most satisfactory position according to the isometric test.
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Articulação do Joelho , Tíbia , Humanos , Fenômenos Biomecânicos , Articulação do Joelho/cirurgia , Tíbia/cirurgia , Fêmur/cirurgia , Amplitude de Movimento Articular , Cadáver , LigamentosRESUMO
Calcium acts as a secondary messenger in plants and is essential for plant growth and development. However, studies on the pathway of aroma synthesis in 'Nanguo' pear (Pyrus ussriensis Maxim.) are scarce. In this study, a bioinformatics analysis of transcriptomic data from calcium-treated 'Nanguo' pear was performed, which identified two fatty acid desaturases, PuFAD2 and PuFAD3, and eight AP2/ERF transcription factors, all exhibiting the same expression patterns. Transient expression experiments showed overexpression of PuFAD2 and PuFAD3 significantly increased the levels of aromatic substrates linoleic acid, hexanal, linolenic acid, and (E)-2-hexenal, but RNAi (RNA interference) had the opposite expression. Promoter sequences analysis revealed that PuFAD2 and PuFAD3 have ERE (estrogen response element) motifs on their promoters. The strongest activation of PuFAD2 by PuERF008 was verified using a dual-luciferase reporting system. Additionally, yeast one-hybrid and electrophoretic mobility shift assays revealed PuERF008 could active PuFAD2. Transient overexpression and RNAi analyses of PuERF008 showed a strong correlation with the expression of PuFAD2. This study provides insights into the process of aroma biosynthesis in 'Nanguo' pear and offers a theoretical basis for elucidating the role of calcium signaling in aroma synthesis.
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Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Pyrus , Pyrus/metabolismo , Pyrus/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Sinalização do Cálcio , Ácidos Graxos/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Regiões Promotoras Genéticas/genética , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Dessaturases/genética , Cálcio/metabolismo , OdorantesRESUMO
OBJECTIVE: Arthroscopic release is effective for patients with shoulder stiffness, but the traditional inside-out procedure cannot effectively alleviate the mobility of some severe stiff shoulder and even cause itrogenic injuries sometimes. The aim of this study is to evaluate the clinical efficacy and advantages of a modified outside-in shoulder release approach for severe shoulder stiffness. METHODS: Included in this retrospective study were 15 patients (five male and 10 female) with severe shoulder stiffness who underwent modified outside-in shoulder release surgery at our hospital between June 2019 and March 2021. Of them, 10 patients had a primary frozen shoulder and five had secondary shoulder stiffness, involving the right shoulder in six cases and the left shoulder in nine cases. The mean age of the 15 patients was 56.7 (34-69) years. The patients were instructed to exercise passively from second-day post-operation and enhance the rehabilitation exercise gradually. All patients received a range of motion (ROM) examination before and after surgery. The American Shoulder and Elbow Surgeon's Score (ASES), Constant Score (CS), and Visual Analog Scale (VAS) score for pain were recorded. All data were tested by normal distribution first and then by paired T test, otherwise by Wilcoxon rank sum test. RESULTS: The mean follow-up period was 18.2 (12-33) months. Compared with the preoperative value, the mean ASES score at the final follow-up improved from 38.4 ± 7.37 to 88.13 ± 6.33 points; the mean CS score from 43.27 ± 6.71 to 78.74 ± 6.93 points; the mean VAS score from 5.07 ± 1.03 to 0.81 ± 0.83 points; forward flexion from 81.93° ± 11.45° to 156.73° ± 9.12°; abduction from 65.93° ± 16.82° to 144.80° ± 8.83°; neutral external rotation from 13.53° ± 10.38° to 51.20° ± 4.77°; internal rotation from the buttock to waist (L3), all showing a significant difference (P < 0.0001). No serious complication was observed in any patient during the postoperative follow-up periods. CONCLUSION: The present study has demonstrated that the modified arthroscopic outside-in shoulder release approach can improve ROM of patients and alleviate pain effectively, proving it to be an appropriate surgical option for the treatment of severe shoulder stiffness.
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Artropatias , Articulação do Ombro , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Ombro , Estudos Retrospectivos , Articulação do Ombro/cirurgia , Artroscopia/métodos , Liberação da Cápsula Articular/métodos , Resultado do Tratamento , Amplitude de Movimento ArticularRESUMO
Hydrogen sulfide (H2S) is involved in multiple processes during plant growth and development. D-cysteine desulfhydrase (DCD) can produce H2S with D-cysteine as the substrate; however, the potential developmental roles of DCD have not been explored during the tomato lifecycle. In the present study, SlDCD2 showed increasing expression during fruit ripening. Compared with the control fruits, the silencing of SlDCD2 by pTRV2-SlDCD2 accelerated fruit ripening. A SlDCD2 gene-edited mutant was constructed by CRISPR/Cas9 transformation, and the mutant exhibited accelerated fruit ripening, decreased H2S release, higher total cysteine and ethylene contents, enhanced chlorophyll degradation and increased carotenoid accumulation. Additionally, the expression of multiple ripening-related genes, including NYC1, PAO, SGR1, PDS, PSY1, ACO1, ACS2, E4, CEL2, and EXP was enhanced during the dcd2 mutant tomato fruit ripening. Compared with the wild-type fruits, SlDCD2 mutation induced H2O2 and malondialdehyde (MDA) accumulation in fruits, which led to an imbalance in reactive oxygen species (ROS) metabolism. A correlation analysis indicated that H2O2 content was strongly positively correlated with carotenoids content, ethylene content and ripening-related gene expression and negatively correlated with the chlorophyll content. Additionally, the dcd2 mutant showed earlier leaf senescence, which may be due to disturbed ROS homeostasis. In short, our findings show that SlDCD2 is involved in H2S generation and that the reduction in endogenous H2S production in the dcd2 mutant causes accelerated fruit ripening and premature leaf senescence. Additionally, decreased H2S in the dcd2 mutant causes excessive H2O2 accumulation and increased ethylene release, suggesting a role of H2S and SlDCD2 in modulating ROS homeostasis and ethylene biosynthesis.
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OBJECTIVES: To evaluate the efficacy and safety of short-term low-dose intravenous haloperidol for delirium prevention in critically ill elderly patients after noncardiac surgery. DESIGN: Prospective, randomized, double-blind, and placebo-controlled trial in two centers. SETTING: Intensive care units of two large tertiary teaching hospitals. PATIENTS: Four hundred fifty-seven patients 65 yrs or older who were admitted to the intensive care unit after noncardiac surgery. INTERVENTION: Haloperidol (0.5 mg intravenous bolus injection followed by continuous infusion at a rate of 0.1 mg/h for 12 hrs; n = 229) or placebo (n = 228) was randomly administered from intensive care unit admission. MEASURES: The primary end point was the incidence of delirium within the first 7 days after surgery. Secondary end points included time to onset of delirium, number of delirium-free days, length of intensive care unit stay, all-cause 28-day mortality, and adverse events. Delirium was assessed using the confusion assessment method for the intensive care unit. RESULTS: The incidence of delirium during the first 7 days after surgery was 15.3% (35/229) in the haloperidol group and 23.2% (53/228) in the control group (p = .031). The mean time to onset of delirium and the mean number of delirium-free days were significantly longer (6.2 days [95% confidence interval 5.9-6.4] vs. 5.7 days [95% confidence interval 5.4-6.0]; p = .021; and 6.8 ± 0.5 days vs. 6.7 ± 0.8 days; p = .027, respectively), whereas the median length of intensive care unit stay was significantly shorter (21.3 hrs [95% confidence interval 20.3-22.2] vs. 23.0 hrs [95% confidence interval 20.9-25.1]; p = .024) in the haloperidol group than in the control group. There was no significant difference with regard to all-cause 28-day mortality between the two groups (0.9% [2/229] vs. 2.6% [6/228]; p = .175). No drug-related side effects were documented. CONCLUSIONS: For elderly patients admitted to intensive care unit after noncardiac surgery, short-term prophylactic administration of low-dose intravenous haloperidol significantly decreased the incidence of postoperative delirium. The therapy was well-tolerated.
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Antipsicóticos/uso terapêutico , Delírio/epidemiologia , Delírio/prevenção & controle , Haloperidol/uso terapêutico , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/prevenção & controle , Idoso , Método Duplo-Cego , Feminino , Humanos , Incidência , Masculino , Estudos ProspectivosRESUMO
OBJECTIVE: To determine the treatment effect of an optimized resuscitation strategy on prognosis of patients with septic shock. METHODS: A systematic review of MEDLINE, the Cochrane Library, EMBASE, CBM, CNKI and other sources using a comprehensive strategy was conducted. All randomized controlled trials (RCTs) of treatment of severe sepsis and septic shock were included. The patients were randomized into treatment group and control group. Preplanned subgroup analysis required studies to be categorized based on early (goal-directed therapy accomplished within 6-24 hours) vs. late (initiated after 24 hours or unknown time of resuscitation implementation). Two authors independently extracted data and assessed study quality using standardized methods. Consensus was reached by conference. The Cochrance Collaboration's software RevMan 5.0 was used for data analysis. RESULTS: Eleven studies were included in the final analysis, providing a sample of 2066 patients. The combined results demonstrated a decrease in mortality [odds ratio (OR) = 0.63, 95% confidence interval (95%CI) 0.48-0.84, P = 0.002]; however, there was statistically significant heterogeneity [P = 0.01, I(2) = 55%]. Among the early optimal resuscitation studies (n = 8) there was minimal heterogeneity [P = 0.44, I(2) = 0%] and a significant decrease in mortality (OR = 0.53, 95%CI 0.42-0.66, P < 0.00001) and incidence of multiple organ dysfunction syndrome (MODS, OR = 0.61, 95%CI 0.43-0.85, P = 0.004). For the late resuscitation studies (n = 3), there was statistically significant difference [P = 0.05, I (2)=66%], and there was no significant effect on mortality (OR = 0.89, 95%CI 0.52-1.54, P = 0.68) and incidence of MODS (OR = 0.43, 95%CI 0.17-1.08, P = 0.07). CONCLUSION: This meta-analysis found that the application of an early optimal resuscitation strategy to patients with septic shock imparts a significant reduction in mortality and incidence of MODS.
Assuntos
Ressuscitação/métodos , Choque Séptico/terapia , Hidratação , Humanos , Prognóstico , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do TratamentoRESUMO
OBJECTIVE: To compare the effect of albumin as a resuscitation fluid with other fluids in lowering the mortality of patients with sepsis. METHODS: By searching MEDLINE, Embase, Cochrane Central Registration of Controlled Trials databases, the metaRegister of Controlled Trials, the Medical Editors Trial Amnesty Register, and retrieval of the randomized controlled trial (RCT) literature to compare the result of resuscitation using albumin-containing fluid and other fluids. The study population included adult patients who were diagnosed to have sepsis, and the patients with sepsis who were studied as subgroup. The RevMan 5.0 software was used for Meta-analysis, and the main outcome was the mortality of the hospitalized patients. RESULTS: In the 14 RCTs, 1729 patients received the albumin-containing fluid resuscitation or resuscitation with other fluids. It was found that the patients with sepsis were the only research objects in five studies, and in other nine studies patients with sepsis were studied as subgroup. P = 0.98, I (2)=0%, i.e. no heterogenicity, and the fixed effect model was used for combining results. There was no evident difference between the group of patients with sepsis resuscitated by albumin-containing fluids and other fluids [odds ratio (OR) was 0.87, 95% confidence interval (95%CI) 0.71-1.07, P = 0.18]. The pooled OR of resuscitation using high concentration albumin solution (20%) was 1.11, 95%CI 0.71-1.73, P = 0.65, the pooled OR of resuscitation using low concentration albumin solution (4%, 5%) was 0.82, 95%CI 0.65-1.03, P = 0.09. Resuscitation with different concentration of albumin-containing fluids was identical with the whole research results. After rejecting six articles of Boldt and other authors, and also saline versus albumin fluid evaluation (SAFE) study, the sensitivity analysis of the study was performed in order to check whether the data produced decisive significance to the whole research results or not. The whole results did not change after their rejection(Boldt studies were rejected:OR 0.82, 95%CI 0.65-1.02, P = 0.08; SAFE study was rejected: OR 1.05, 95%CI 0.71-1.55, P = 0.82). Therefore the analysis results were satisfactory. CONCLUSION: The Meta-analysis shows that by using albumin-containing fluids for resuscitation can not lower the mortality of sepsis as compared with other fluids.
Assuntos
Albuminas/uso terapêutico , Hidratação/métodos , Sepse/terapia , Humanos , Prognóstico , Ensaios Clínicos Controlados Aleatórios como Assunto , RessuscitaçãoRESUMO
OBJECTIVE: To demonstrate the pharmacokinetic profile of meropenem when administered by 3-hour infusion in patients undergoing continuous veno-venous hemofiltration (CVVH). METHODS: The study was conducted in 10 patients, who were treated with CVVH. Each subject received meropenem in 3-hour infusion of 500 mg every 6 hours. Blood samples were collected before infusion (0 hour) and 0.25, 0.5, 1, 1.5, 2, 3, 4, 5, 6 hours (just before the infusion of the next dose) after the beginning of the fourth infusion. The concentrations of meropenem in plasma were measured by high-performance liquid chromatography method, and mean serum meropenem concentration-time curve was plotted. RESULTS: Peak plasma drug concentrations measured 3 hours post-infusion were (25.05 ± 5.64) mg/L, and trough levels after 6 hours of infusion were (13.03 ± 3.01) mg/L. The area under the plasma concentration-time curve (AUC) was (118.42 ± 26.78) mg x h⻹ x L⻲. The elimination half-life (T1/2) was (3.74 ± 0.55) hours. The mean residence time (MRT) was (4.99 ± 0.84) hours. The volume of distribution (Vb) was (22.85 ± 9.85) L and clearance of meropenem (CL) was (4.49 ± 1.32) L/h. The percentage of time that the serum drug concentration was above the minimum inhibitory concentration (MIC) accounting for the interval time of infusion (%T>MIC) was 100% (MIC 8 mg/L) in all the 10 patients. CONCLUSION: Based on these data, we concluded that satisfactory pharmacodynamic parameters could be attained in CVVH patients treated with meropenem by a prolonged infusion time of 3 hours with a dosage of 500 mg for every 6 hours.
Assuntos
Hemofiltração , Tienamicinas/administração & dosagem , Tienamicinas/farmacocinética , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/terapia , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Feminino , Humanos , Infusões Intravenosas , Masculino , Meropeném , Pessoa de Meia-Idade , Estudos Prospectivos , Sepse/metabolismo , Sepse/terapiaRESUMO
OBJECTIVE: To identify the risk factors of prolonged intensive care unit (ICU) stay of critically ill obstetric patients. METHODS: A retrospective analysis of cases of critically ill obstetric patients admitted to the ICUs of Peking University Third Hospital, Capital Medical University Affiliated Beijing Chaoyang Hospital, and PLA 306 Hospital from January 1st 2006 to December 31st 2010 was made. Data included demographics, causes of critical illness or complications that prompted ICU admission, the acute physiology and chronic health evaluation II (APACHEII) scores, the time intervals between onset of acute symptoms and ICU admission, laboratory test results, treatment measures, length of ICU stay and the final maternal mortality. Data were used to identify univariate and multivariate predictors for prolonged ICU stay. RESULTS: During the 5-year period there were 207 obstetric patients [mean age (31.74±2.32) years old, mean gestational age (34.86±4.72) weeks] were transferred to the ICU for critical care (42 ICU admissions per 10 000 deliveries), and among them 4 women died (mortality rate 1.93%). The pathogenesis of the cases could be divided into direct obstetric pathologies (n=138) and indirect or coincidental pathologies (n=69). The most common obstetric causes of admission were massive postpartum haemorrhage (n=42, 20.29%) and pregnancy associated hypertension (n=36, 17.39%), followed by acute fatty liver of pregnancy (AFLP, n=27, 13.04%), obstetric disseminated intravascular coagulation (DIC, n=23, 11.11%). The most common non obstetric causes of admission were acute heart failure (n=26, 12.56%) and acute respiratory failure (n=22, 10.63%), followed by severe acute pancreatitis (SAP, n=11, 5.31%). The incidence of prolonged ICU stay (ICU stay >3 days) was 52.66% (n=109). Univariate predictors of prolonged ICU stay included inadvertent antenatal care, high serum creatinine, abnormal international normalized ratio of prothrombin time (INR), abnormal oxygenation index (PaO(2)/FiO(2)), AFLP, obstetric DIC, SAP, use of mechanical ventilation, indications for inotropic support, and blood purification, and >24 hour interval between onset of acute symptoms and ICU admission. Multivariate modeling identified that inadvertent and irregular antenatal care [odds ratio (OR) 1.68, 95% confidence interval (95%CI) 1.14-2.69, P=0.011], PaO(2)/FiO(2) (OR 4.73, 95%CI 1.46-11.37, P=0.013), AFLP (OR3.21, 95%CI 1.13-4.76, P=0.026), DIC (OR 2.73, 95%CI 1.28-4.02, P=0.018), SAP (OR 4.78, 95%CI 1.83-7.42, P=0.021 ), indications for inotropic support (OR 1.96, 95%CI 1.24-3.15, P=0.001), blood purification (OR 11.02, 95%CI 3.04-58.02, P=0.015) and >24 hour interval between onset of acute symptoms and ICU admission (OR 2.04, 95%CI 1.21-4.25, P<0.001) were the independent predictors for prolonged ICU stay. CONCLUSION: The incidence of prolonged ICU stay is high for critically ill obstetric patients. Patients with identified multivariate predictors carry a high risk of prolonged ICU stay, they may benefit from enhanced regular antenatal care, prevention of critical obstetric and medical complications, shortening the interval between onset of acute symptoms and ICU admission and strengthening the support of organ function.
Assuntos
Estado Terminal , Unidades de Terapia Intensiva , Tempo de Internação , Adulto , Feminino , Humanos , Gravidez , Estudos Retrospectivos , Fatores de Risco , Adulto JovemRESUMO
Purple-fleshed sweetpotato (Ipomoea batatasï¼L.ï¼Lam.) is rich in anthocyanins. R2R3-type MYB transcription factorsï¼TFsï¼with EAR motifs inhibiting anthocyanin biosynthesis have been reported, and there is still a lack of information on how mutations in the EAR motifs of MYBs affect anthocyanin accumulation. In this study, we obtained three IbMYB44 TFs by bioinformatics. Among these TFs, IbMYB44.1, IbMYB44.3 with a complete EAR motif and IbMYB44.2 with a single amino acid mutant in the EAR motif caused an amino acid substitution from leucine to valine. RT-qPCR analysis showed that IbMYB44s was expressed at lower levels in the purple-fleshed sweetpotato than in nonpurple-fleshed sweetpotato (P < 0.01). Transient expression assays showed that the inhibitory effect of IbMYB44.1/3 was stronger than IbMYB44.2 in tobacco leaves and red-skinned pears. RT-qPCR analysis further proved that IbMYB44.1/3 significantly inhibited the expression of anthocyanin biosynthesis-related genes compared with IbMYB44.2 in tobacco leaves and red-skinned pears. A dual luciferase reporter assay showed that IbMYB44s cannot directly activate the IbANS promoter, and the result was also verified by yeast one-hybrid (Y1H) experiments. Moreover, we identified the interaction of IbMYB340 with IbMYB44.1, IbMYB44.2 and IbMYB44.3 via yeast two-hybrid (Y2H) assays. Thus, IbMYB44.1/3 could interact with IbMYB340 to negatively regulate anthocyanin biosynthesis. This study enriched the regulatory network of anthocyanins and also provided a theoretical basis for a single amino acid mutant from leucine to valine in the EAR motif of IbMYB44.2 affecting anthocyanin biosynthesis in the purple-fleshed sweetpotato.
Assuntos
Ipomoea batatas , Aminoácidos , Antocianinas , Regulação da Expressão Gênica de Plantas , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismoRESUMO
Sweet potato decays easily due to its high respiration rate and reactive oxygen species (ROS) accumulation during postharvest storage. In this study, we explored the relationship between antioxidant capacity in leaves and storage properties in different sweet potato cultivars, the tuberous roots of 10 sweet potato cultivars were used as the experimental materials to analyze the storage property during storage at 11-15°C. According to the decay percentage after 290 days of storage, Xu 32 was defined as a storage-tolerant cultivar (rot percentage less than 25%); Xu 55-2, Z 15-1, Shangshu 19, Yushu, and Zhezi 3 as above-moderate storage-tolerant cultivars (rot percentage ranging from 25 to 50%); Sushu 16, Yanshu 5, and Hanzi as medium-storable cultivars (rot percentage 50-75%); and Yan 25 as a storage-sensitive cultivar (rot percentage greater than 75%). Meanwhile, analysis of the α-amylase activity in root tubers of the 10 sweet potato cultivars during storage indicated that α-amylase activity was lowest in the storage-tolerant cultivar Xu 32 and highest in the storage-sensitive cultivar Yan 25. Evaluation of antioxidant enzyme activities and ROS content in the leaves of these 10 cultivars demonstrated that cultivar Xu 32, which showed the best storage property, had higher antioxidant enzyme activity [superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD)] but lower lipoxygenase (LOX) activity, hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents, and superoxide anion radical (O2â -) production rates compared with those of the storage-sensitive cultivar Yan 25 and the medium-storability cultivars Hanzi, Yanshu 5, and Sushu 16. Additionally, principal component analysis (PCA) suggested that sweet potato cultivars with different storage properties were clustered separately. Correlation and heat map analysis further indicated that CAT, APX, POD, and SOD activities were negatively correlated with α-amylase activity, while LOX activity and MDA and H2O2 contents were negatively correlated with the storage property of sweet potato. Combined, our findings revealed that storage property is highly correlated with antioxidant capacity in sweet potato leaves and negatively correlated with α-amylase activity in tuberous roots, which provides a convenient means for the screening of storage-tolerant sweet potato cultivars.