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BACKGROUND: Skin flap transplantation is used to effectively reconstruct defects of the hand and foot skin and soft tissues. We here investigated the effect of the PF127/bleomycin (BLM) hydrogel on the extracellular matrix (ECM) remodeling of skin flaps and the underlying mechanism, thereby providing a new reference point for personalized flap modification and overcoming abrasion resistance- and stability-associated difficulties. METHODS: The appropriate PF127/BLM concentration was selected based on the gelation time and drug release curve. Migration assays, scratch assays, and live/dead staining were conducted to verify the effect of PF127/BLM on human skin fibroblasts (HSFs). The effects of PF127/BLM on the ECM were assessed through hematoxylin and eosin and Masson staining. Additionally, we examined the expression of ECM remodeling-related genes and proteins involved in their associated signaling pathway. Finally, the effects of PF127/BLM on organ fibrosis and toxicity to liver and kidney functions were assessed in mice. RESULTS: A 25% PF127/BLM hydrogel was selected as the study concentration. PF127/BLM augmented HSF chemotaxis and proliferation. Furthermore, PF127/BLM promoted subcutaneous ECM remodeling and fibrosis, increased the flap dermis thickness, and reduced the toxic side effects of BLM on liver/lung fibrosis and liver/kidney function. Additional studies confirmed that the PF127/BLM-mediated regulation of ECM remodeling in skin flaps was associated with TGFß-Col signaling pathway activation. CONCLUSION: The PF127/BLM hydrogel promoted subcutaneous ECM remodeling and fibrosis, which aided the construction of personalized flaps through the TGFß-Col signaling pathway, with decreased hepatic, pulmonary, and renal toxicities.
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Bleomicina , Matriz Extracelular , Hidrogéis , Transdução de Sinais , Fator de Crescimento Transformador beta , Matriz Extracelular/metabolismo , Matriz Extracelular/efeitos dos fármacos , Animais , Fator de Crescimento Transformador beta/metabolismo , Camundongos , Humanos , Transdução de Sinais/efeitos dos fármacos , Bleomicina/toxicidade , Bleomicina/farmacologia , Fibrose , Retalhos Cirúrgicos/patologia , Masculino , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/patologia , Colágeno , Medicina de Precisão/métodosRESUMO
We aimed to obtain the optimal formula for human milk fat substitute (HMFS) through a combination of software and an evaluation model and further verify its practicability through an animal experiment. The results showed that a total of 33 fatty acid (FA) and 63 triglyceride (TAG) molecular species were detected in vegetable oils. Palmitic acid, oleic acid, linoleic acid, 18:1/16:0/18:1, 18:2/16:0/18:2, 18:1/18:1/18:1 and 18:1/18:2/18:1, were the main molecular species among the FAs and TAGs in the vegetable oils. Based on the HMFS evaluation model, the optimal mixed vegetable oil formula was blended with 21.3% palm oil, 2.8% linseed oil, 2.6% soybean oil, 29.9% rapeseed oil and 43.4% maize oil, with the highest score of 83.146. Moreover, there was no difference in the weight, blood routine indices or calcium and magnesium concentrations in the feces of the mice between the homemade mixed vegetable oil (HMVO) group and the commercial mixed vegetable oil (CMVO) group, while nervonic acid (C24:1) and octanoic acid (C8:0) were absorbed easily in the HMVO group. Therefore, these results demonstrate that the mixing of the different vegetable oils was feasible via a combination of computer software and an evaluation model and provided a new way to produce HMFS.
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Substitutos da Gordura , Ácidos Graxos , Leite Humano , Óleos de Plantas , Software , Triglicerídeos , Humanos , Animais , Óleos de Plantas/química , Ácidos Graxos/química , Leite Humano/química , Camundongos , Triglicerídeos/química , Substitutos da Gordura/química , Óleo de Palmeira/química , Óleo de Soja/química , Óleo de Semente do Linho/química , Óleo de Brassica napus/química , Óleo de Milho/química , Caprilatos/química , Ácido Palmítico/química , Ácido Oleico/químicaRESUMO
BACKGROUND: Polydatin has shown considerable pharmacological activities in ischemia-reperfusion injuries of various organs. However, its effects and mechanisms in spinal cord ischemia-reperfusion injury have not been fully established. In this study, the mechanisms of polydatin against spinal cord ischemia-reperfusion injury were investigated via network pharmacology, molecular docking and molecular dynamics simulation. METHODS: Spinal cord ischemia-reperfusion injury-related targets were obtained from the GeneCards database, while polydatin-related action targets were obtained from the CTD and SwissTarget databases. A protein-protein interaction network of potential targets was constructed using the String platform. After selecting the potential key targets, GO functional enrichment and KEGG pathway enrichment analyses were performed via the Metascape database, and a network map of "drug-target-pathway-disease" constructed. The relationships between polydatin and various key targets were assessed via molecular docking. Molecular dynamics simulation was conducted for optimal core protein-compound complexes obtained by molecular docking. RESULTS: Topological analysis of the PPI network revealed 14 core targets. GO functional enrichment analysis revealed that 435 biological processes, 12 cell components and 29 molecular functions were enriched while KEGG pathway enrichment analysis revealed 91 enriched signaling pathways. Molecular docking showed that polydatin had the highest binding affinity for MAPK3, suggesting that MAPK3 is a key target of polydatin against spinal cord ischemia-reperfusion injury. Molecular dynamics simulations revealed good binding abilities between polydatin and MAPK3. CONCLUSIONS: Polydatin exerts its effects on spinal cord ischemia-reperfusion injury through multiple targets and pathways. MAPK3 may be a key target of polydatin in spinal cord ischemia-reperfusion injury.
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Simulação de Dinâmica Molecular , Traumatismo por Reperfusão , Medula Espinal , Humanos , Simulação de Acoplamento Molecular , Farmacologia em Rede , Traumatismo por Reperfusão/tratamento farmacológicoRESUMO
In this study, we built classification models using machine learning techniques to predict the bioactivity of non-covalent inhibitors of Bruton's tyrosine kinase (BTK) and to provide interpretable and transparent explanations for these predictions. To achieve this, we gathered data on BTK inhibitors from the Reaxys and ChEMBL databases, removing compounds with covalent bonds and duplicates to obtain a dataset of 3895 inhibitors of non-covalent. These inhibitors were characterized using MACCS fingerprints and Morgan fingerprints, and four traditional machine learning algorithms (decision trees (DT), random forests (RF), support vector machines (SVM), and extreme gradient boosting (XGBoost)) were used to build 16 classification models. In addition, four deep learning models were developed using deep neural networks (DNN). The best model, Model D_4, which was built using XGBoost and MACCS fingerprints, achieved an accuracy of 94.1% and a Matthews correlation coefficient (MCC) of 0.75 on the test set. To provide interpretable explanations, we employed the SHAP method to decompose the predicted values into the contributions of each feature. We also used K-means dimensionality reduction and hierarchical clustering to visualize the clustering effects of molecular structures of the inhibitors. The results of this study were validated using crystal structures, and we found that the interaction between the BTK amino acid residue and the important features of clustered scaffold was consistent with the known properties of the complex crystal structures. Overall, our models demonstrated high predictive ability and a qualitative model can be converted to a quantitative model to some extent by SHAP, making them valuable for guiding the design of new BTK inhibitors with desired activity.
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FMS-like tyrosine kinase 3 (FLT3) is a type III receptor tyrosine kinase, which is an important target for anti-cancer therapy. In this work, we conducted a structure-activity relationship (SAR) study on 3867 FLT3 inhibitors we collected. MACCS fingerprints, ECFP4 fingerprints, and TT fingerprints were used to represent the inhibitors in the dataset. A total of 36 classification models were built based on support vector machine (SVM), random forest (RF), eXtreme Gradient Boosting (XGBoost), and deep neural networks (DNN) algorithms. Model 3D_3 built by deep neural networks (DNN) and TT fingerprints performed best on the test set with the highest prediction accuracy of 85.83% and Matthews correlation coefficient (MCC) of 0.72 and also performed well on the external test set. In addition, we clustered 3867 inhibitors into 11 subsets by the K-Means algorithm to figure out the structural characteristics of the reported FLT3 inhibitors. Finally, we analyzed the SAR of FLT3 inhibitors by RF algorithm based on ECFP4 fingerprints. The results showed that 2-aminopyrimidine, 1-ethylpiperidine,2,4-bis(methylamino)pyrimidine, amino-aromatic heterocycle, [(2E)-but-2-enyl]dimethylamine, but-2-enyl, and alkynyl were typical fragments among highly active inhibitors. Besides, three scaffolds in Subset_A (Subset 4), Subset_B, and Subset_C showed a significant relationship to inhibition activity targeting FLT3.
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BACKGROUND: Vascular malformations are common but complicated types of disease in infants, with unclear causes and lack of effective prevention. The symptoms usually do not disappear and tend to progress without medical intervention. It is extremely necessary to choose correct treatment options for different types of vascular malformations. A large number of studies have confirmed that sclerotherapy has a tendency to become the first-line treatment in near future, but it is also associated with mild or severe complications. Furthermore, to our knowledge, the serious adverse event of progressive limb necrosis has not been systematically analyzed and reported in the literature. CASE PRESENTATION: Three cases (two females and one male) were presented who were all diagnosed as vascular malformations and were treated by several sessions of interventional sclerotherapy. Their previous medical records showed the use of several sclerosants in different sessions including Polidocanol and Bleomycin. The sign of limb necrosis did not occur during the first sclerotherapy, but after the second and third sessions. Furthermore, the short-term symptomatic treatment could improve the necrosis syndrome, but could not change the outcome of amputation. CONCLUSION: Sclerotherapy undoubtedly tends to be the first-line treatment in near future, but the adverse reactions still remain major challenges. Awareness of progressive limb necrosis after sclerotherapy and timely management by experts in centers of experience of this complication can avoid amputation.
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Escleroterapia , Malformações Vasculares , Lactente , Feminino , Humanos , Masculino , Escleroterapia/efeitos adversos , Resultado do Tratamento , Estudos Retrospectivos , Soluções Esclerosantes/efeitos adversos , Malformações Vasculares/complicações , Malformações Vasculares/terapia , Malformações Vasculares/diagnósticoRESUMO
In this study, we aimed to detect the physicochemical properties of distilled products (residue and distillate) obtained from anhydrous milk fat (AMF) and its dry fractionation products (liquid and solid fractions at 25°C [25 L and 25 S]). The results showed that the saturated fatty acids and low- and medium molecular-weight triglycerides were easily accumulated in the distillate, and the percentage of unsaturated fatty acid and high molecular-weight triglycerides in the residue were higher, and these components in 25 S and 25 L were influenced more significantly than those in the AMF. In addition, the distillate had larger melting ranges in comparison with the distilled substrate, while the melting ranges of residue was smaller. The triglycerides were presented as the mixture crystal forms (α, ß', and ß crystal) in 25 S, AMF, and their distilling products, and it was transformed gradually to a single form as the increasing of distilling temperature. Moreover, the accumulated pattern of triglycerides was double chain length in 25 S, AMF, and their distilling products. These results provide a new approach to obtain the milk fat fractions with different properties, and the findings of this study enrich the theoretical basis of milk fat separation in practical production.
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Ácidos Graxos , Leite , Animais , Ácidos Graxos/análise , Triglicerídeos/análise , Leite/química , Destilação , Fracionamento QuímicoRESUMO
Kestoses, the smallest fructooligosaccharides, are trisaccharides composed of a fructose molecule and a sucrose molecule linked by either ß-(2,1) or ß-(2,6) linkage. 1-kestose, 6-kestose and neokestose are the three types of kestoses occurring in nature. As the main kind of fructooligosaccharide, kestoses share similar physiological effects with other fructooligosaccharides, and they have recently been determined to show more notable effects in promoting the growth of probiotics including Faecalibacterium prausnitzii and Bifidobacterium than those of other fructooligosaccharides. Kestoses exist in many plants, but the relatively low content and the isolation and purification are the main barriers limiting their industrial application. The production of kestoses by enzymatic biosynthesis and microbial fermentation has the potential to facilitate its production and industrial use. In this article, the recent advances in the research of kestoses were overviewed, including those studying their functions and production. Kestose-producing enzymes were introduced in detail, and microbial production and fermentation optimization techniques for enhancing the yield of kestoses were addressed. ß-Fructofuranosidase is the main one used to produce kestoses because of the extensive range of microbial sources. Therefore, the production of kestoses by microorganisms containing ß-fructofuranosidase has also been reviewed. However, few molecular modification studies have attempted to change the production profile of some enzymes and improve the yield of kestoses, which is a topic that should garner more attention. Additionally, the production of kestoses using food-grade microorganisms may be beneficial to their application in the food industry.
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Biotecnologia , Oligossacarídeos , Biotecnologia/tendências , Enzimas/metabolismo , Fermentação , Microbiologia de Alimentos , Oligossacarídeos/metabolismoRESUMO
OBJECTIVES: The polyfoliate anterolateral thigh perforator flap needs to dissect two or more perforators, which is an ideal choice for repairing wide and irregular wounds. However, the uncertainty of perforating vessels restricts the development of this operation. This study discusses the feasibility and clinical efficacy of the polyfoliate anterolateral thigh perforator flap with single-perforator. METHODS: Fifteen patients with skin and soft tissue defects in extremities, were treated with polyfoliate anterolateral thigh perforator flap with single-perforator. Based on the perforator detected by Doppler ultrasound or color Doppler ultrasonography before operation, a polyfoliate anterolateral thigh perforator flap with single-perforator was designed. The perforating point of perforator was near the boundary of the skin paddle. Following the perforating vessels and vascular pedicles free, the vessels in the deep layer of the superficial fascia were meticulously free under the microscope. After obtaining the appropriate length, the skinpaddles were separated and recombined. After confirming the blood supply of flap, the vascular pedicle was ligated and transplanted to the recipient area. RESULTS: In 15 cases, the area of the flap was 8.0 cm×5.0 cm+6.0 cm×5.5 cm to 16.0 cm× 9.5 cm+24.0 cm×9.0 cm. All flaps survived well without necrosis and had a satisfactory appearance. The donor area was closed directly. The patients were followed up for 3 to 12 months, with an average of 6 months. The skin flaps were normal in color and good in texture. CONCLUSIONS: It's a better method to repair the skin and soft tissue defects in extremities by the polyfoliate anterolateral thigh perforator flap with single-perforator because only one perforator needs to be dissected, a group of blood vessels need to be anastomosed, and only one donor area needs to be sacrificed.
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Retalho Perfurante , Procedimentos de Cirurgia Plástica , Lesões dos Tecidos Moles , Humanos , Transplante de Pele , Lesões dos Tecidos Moles/cirurgia , Coxa da Perna/cirurgia , Resultado do TratamentoRESUMO
BACKGROUND: Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metabolites toward CRC cells is not clear. Therefore, the aim of this study was to assess the inhibitory mechanism of cell-free supernatants (CFSs) of L. rhamnosus GG, L. casei M3, and L. plantarum YYC-3 on metastasis of CRC cells. RESULTS: YYC-3 CFS showed the highest inhibitory effect on CRC cell growth, invasion and migration, and inhibited MMP2, MMP9, and VEGFA gene and protein expression, and protein secretion. Furthermore, it suppressed the activities of MMPs by gelatin zymography. Moreover, the effective compounds in these CFSs were analyzed by Q Exactive Focus liquid chromatography-mass spectrometry. CONCLUSIONS: Our results showed that metabolite secretions of YYC-3 may inhibited cell metastasis by downregulating the VEGF/MMPs signaling pathway. These data suggest that treatment of CRC cells with metabolites from L. plantarum YYC-3 may reduce colon cancer metastasis.
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Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/microbiologia , Lactobacillus/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Células CACO-2 , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Células HT29 , Humanos , Lacticaseibacillus casei/metabolismo , Lactobacillus plantarum/metabolismo , Lacticaseibacillus rhamnosus/metabolismo , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Metástase Neoplásica/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
The probiotic efficacy and fermentative ability of Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus), a widely used probiotic, is majorly affected by its acid tolerance. Here, we conducted whole-genome sequencing of the high acid-tolerant L. bulgaricus LJJ stored in the laboratory. Compared with the whole genome of low acid-tolerant strain L. bulgaricus ATCC11842, the results show that 16 candidate acid-tolerant genes may be involved in the regulation of the acid tolerance of L. bulgaricus LJJ. Association analysis of candidate acid-tolerant genes and acid-tolerant traits of different L. bulgaricus strains revealed that the three genes dapA, dapH, and lysC are the main reasons for the strong acid tolerance of L. bulgaricus LJJ. The results of real-time quantitative PCR (RT-qPCR) supported this conclusion. KEGG pathway analysis showed that these three acid-tolerant genes are involved in the synthesis of lysine; the synthesis of lysine may confer L. bulgaricus LJJ strong acid tolerance. This study successfully revealed the acid tolerance mechanism of L. bulgaricus LJJ and provides a theoretical basis for the subsequent selection of strains with high acid tolerance for improved probiotic functions. KEY POINTS: ⢠Three genes are identified as acid-tolerant genes, respectively, lysC, dapA, and dapH. ⢠LysC and dapA are the major key genes in the synthesis of lysine. ⢠The synthesis of lysine may confer L. bulgaricus LJJ strong acid tolerance.
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Lactobacillus delbrueckii , Probióticos , Ácidos/toxicidade , Fermentação , Genômica , Lactobacillus delbrueckii/genéticaRESUMO
PURPOSE: The aim of this study was to demonstrate the viability of the transverse circumflex scapular artery perforator flap (TCSAPF) in children with soft tissue defects of the lower limb. METHODS: In an anatomic study, 25 fresh cadavers were injected with lead oxide-gelatin for spiral computed tomography and 3-dimensional image reconstruction. In a 3-year clinical application study, children with soft tissue defects and exposed tendons and/or bones in the lower limb underwent free-TCSAPF repair of the defect. RESULTS: Perforators from the transverse branch of the circumflex scapular artery were identified in both anatomical and clinical studies. The average external diameter was 0.9 ± 0.3 mm. Each perforator supplied an average area of 63.5 ± 16.8 cm in anatomical. Twenty-one children were included in this group (9 boys, 12 girls, mean age, 6.6 ± 2.7 years). The size of the flaps ranged from 6 to 17 cm × 4.5 to 7 cm (average, 65.3 ± 22.6 cm). The average flap harvesting time was 30.1 ± 8.5 minutes, average operation time was 138.6 ± 31.5 minutes, and average blood loss was 89.5 ± 21.9 mL. The average length of the vessel pedicle was 8.2 ± 2.4 cm. Arterial congestion occurred in one child, 18 hours postoperatively; subsequent re-exploration and great saphenous vein transplantation were successful. Of the 3 children who had bulky flaps, 1 patient underwent defatting. Satisfactory outcomes included good appearance and function of the recipient and donor areas. CONCLUSIONS: The TCSAPF provides high-quality skin and vessel flexibility, providing a reliable blood supply in children. The flap has potential benefits over existing perforator flaps.
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Retalho Perfurante , Procedimentos de Cirurgia Plástica , Lesões dos Tecidos Moles , Artérias/cirurgia , Criança , Pré-Escolar , Feminino , Humanos , Extremidade Inferior/lesões , Extremidade Inferior/cirurgia , Masculino , Transplante de Pele , Lesões dos Tecidos Moles/cirurgiaRESUMO
BACKGROUND: Tumor protein p63 (TP63)-related disorders can be divided into at least six categories, including ectrodactyly-ectodermal dysplasia-cleft lip/palate syndrome 3 (EEC syndrome 3), ankyloblepharon-ectodermal defects-cleft lip/palate syndrome (AEC syndrome), acro-dermo-ungual-lacrimal-tooth syndrome (ADULT syndrome), limb-mammary syndrome (LMS), Rapp-Hodgkin syndrome (RHS) and split-hand/foot malformation 4 (SHFM4), and are all a result of heterozygous mutations of TP63. The phenotypes of TP63-related disorders broadly involve ectodermal dysplasias, acromelic malformation and orofacial cleft. SHFM and hypodontia are prominent clinical manifestations of TP63-related disorders. METHODS: The present study investigated a family with SHFM and hypodontia; determined the sequences of DLX5, WNT8B, WNT10B, BHLHA9, CDH3, DYNC1I1 and FGFR1; and performed single nucleotide polymorphism-array analysis. We detected the mutation by multiple sequence alignments and a bioinformatic prediction. RESULTS: We identified a novel missense mutation of TP63 (c.1010G>T; R337L) in the family without mutations of DLX5, WNT8B, WNT10B, BHLHA9, CDH3, DYNC1I1, FGFR1 and copy number variants causing SHFM. CONCLUSIONS: A mutation of TP63 (c.1010G>T; R337L) leads to SHFM with hypodontia. The identification of this mutation expands the spectrum of known TP63 mutations and also may contribute to novel approaches for the genetic diagnosis and counseling of families with TP63-related disorders.
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Alelos , Substituição de Aminoácidos , Anodontia/diagnóstico , Anodontia/genética , Deformidades Congênitas dos Membros/diagnóstico , Deformidades Congênitas dos Membros/genética , Mutação , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética , Adulto , Criança , Biologia Computacional , Análise Mutacional de DNA , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Masculino , Linhagem , Fenótipo , Polimorfismo de Nucleotídeo Único , Radiografia , SíndromeRESUMO
BACKGROUND: The traditional method of bacterial identification based on 16S rRNA is a widely used and very effective detection method, but this method still has some deficiencies, especially in the identification of closely related strains. A high homology with little differences is mostly observed in the 16S sequence of closely related bacteria, which results in difficulty to distinguish them by 16S rRNA-based detection method. In order to develop a rapid and accurate method of bacterial identification, we studied the possibility of identifying bacteria with other characteristic fragments without the use of 16S rRNA as detection targets. RESULTS: We analyzed the potential of using cas (CRISPR-associated proteins) gene as a target for bacteria detection. We found that certain fragment located in the casx gene was species-specific and could be used as a specific target gene. Based on these fragments, we established a TaqMan MGB Real-time PCR method for detecting bacteria. We found that the method used in this study had the advantages of high sensitivity and good specificity. CONCLUSIONS: The casx gene-based method of bacterial identification could be used as a supplement to the conventional 16 s rRNA-based detection method. This method has an advantage over the 16 s rRNA-based detection method in distinguishing the genetic relationship between closely-related bacteria, such as subgroup bacteria, and can be used as a supplement to the 16 s rRNA-based detection method.
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Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana/métodos , Proteínas Associadas a CRISPR/genética , Lacticaseibacillus casei/isolamento & purificação , Sequência de Bases , Lacticaseibacillus casei/classificação , Lacticaseibacillus casei/genética , Filogenia , Reação em Cadeia da PolimeraseRESUMO
A total of 85 strains of lactic acid bacteria were isolated from corn silage in this study and analyzed in vitro for their cholesterol removal, NPC1L1 protein down-regulation and bile salt deconjugation ability, respectively. Nineteen strains were selected for further analysis for their probiotic potential. Finally, 3 strains showing better probiotic potential were evaluated for their cholesterol-lowering activity in hamsters. The strains showing the greater cholesterol removal and NPC1L1 protein down-regulation activity had no significant effects on serum and hepatic cholesterol levels in hamsters (p > 0.05). However, Lactobacillus plantarum CAAS 18008 (1 × 109 CFU/d) showing the greater bile salt deconjugation ability significantly reduced serum low-density lipoprotein cholesterol, total cholesterol, and hepatic total cholesterol levels by 28.8%, 21.7%, and 30.9%, respectively (p < 0.05). The cholesterol-lowering mechanism was attributed to its bile salt hydrolase activity, which enhanced daily fecal bile acid excretion levels and thereby accelerated new bile acid synthesis from cholesterol in liver. This study demonstrated that the strains showing greater cholesterol removal and NPC1L1 protein down-regulation activity in vitro hardly reveal cholesterol-lowering activity in vivo, whereas the strains showing greater bile salt deconjugation ability in vitro has large potential to decrease serum cholesterol levels in vivo.
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Lactobacillales/isolamento & purificação , Probióticos/isolamento & purificação , Silagem/microbiologia , Zea mays/microbiologia , Animais , Ácidos e Sais Biliares/metabolismo , Colesterol/metabolismo , Colesterol 7-alfa-Hidroxilase , Cricetinae , Fezes/química , Humanos , Lactobacillales/metabolismo , Metabolismo dos Lipídeos , Fígado/metabolismo , Proteínas de Membrana/metabolismo , Proteínas de Membrana Transportadoras , Redes e Vias Metabólicas , Viabilidade Microbiana , Probióticos/metabolismo , Esteroides/química , Esteroides/isolamento & purificaçãoRESUMO
BACKGROUND: Thermostable lipases from microbial sources have been substantially overexpressed in E. coli, however, these enzymes are often produced with low-level enzymatic activity and mainly in the form of inclusion bodies. Several studies have reported that the secretory production of recombinant proteins fused their N-terminus to a signal peptide has been employed to resolve the problem. In general, the feasibility of this approach largely depends on the secretory pathway of signal peptide and the type of target protein to be secreted. This study was performed to compare and optimize signal peptides for efficient secretion of thermostable lipase lipBJ10 from Pseudomonas fluorescens BJ-10. Meanwhile, a comparative study between this method and cytoplasmic secretion was implemented in secreting soluble and active lipases. RESULTS: Fusion expression using six signal peptides, i.e., PelB and five native E. coli signal peptides, as fusion partners produced more soluble and functional recombinant lipBJ10 than non-fusion expression. Recombinant lipBJ10, fused to these six diverse signal peptides, was secreted into the periplasm in E. coli. The total lipase activity in all cases of fusion expression was higher than those in non-fusion expression. The relative activity peaked when lipBJ10 was fused to DsbA, yielding a value 73.3 times greater than that of the non-fusion protein. When DsbA was used as the fusion partner, the highest activity (265.41 U/ml) was achieved with the least formation of inclusion bodies; the other four E. coli signal peptides, to some extent, led to low activity and insoluble inclusion bodies. Therefore, DsbA is the optimal signal peptide partner to fuse with lipBJ10 to efficiently produce soluble and functional protein. CONCLUSION: We found that fusing to these signal peptides, especially that of DsbA, can significantly decrease the formation of inclusion bodies and enhance the function and solubility of lipBJ10 compared to non-fusion lipBJ10. Our results reported here can provide a reference for the high-level expression of other lipases with respect to a possible industrial application.
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Proteínas de Bactérias/química , Escherichia coli/química , Lipase/química , Sinais Direcionadores de Proteínas/fisiologia , Pseudomonas fluorescens/metabolismoRESUMO
BACKGROUND: This study investigates the feasibility and clinical impact of the microdissected thin perforator skin flap strategy on bulky and deformed skin flaps during second-stage revision surgery. METHODS: Seventeen patients were selected and underwent the microdissected thin perforator skin flap technique to treat bulky and deformed skin flaps after free flap reconstruction between October 2013 and October 2015. Perforator vessels were isolated and protected under a microscope. Subdermal fat with a thickness of 4 mm to 7 mm was preserved, and excess adipose tissue was resected. RESULTS: No skin flap necrosis was observed after the operation in all 17 patients, and all wounds healed without complications. Patients were followed up for 3 to 24 months, with an average follow-up time of 10 months. The skin flaps maintain normal color and texture. Both appearance and function of the recipient sites were improved significantly. CONCLUSIONS: The utilization of microdissected thin perforator flap technique to further thin bulky skin flaps at the second stage can be effective in a single operation. The blood supply of all free flaps was preserved, with no evidence of necrosis or healing complications. This technique offers an effective approach for secondary thinning of bulky free flaps.
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Retalho Perfurante/irrigação sanguínea , Procedimentos de Cirurgia Plástica/métodos , Complicações Pós-Operatórias/cirurgia , Transplante de Pele/métodos , Adolescente , Adulto , Criança , Estudos de Viabilidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reoperação , Resultado do Tratamento , Ultrassonografia Doppler , CicatrizaçãoAssuntos
Retalho Perfurante , Coxa da Perna , Angiografia , Catéteres , Computadores , Humanos , Coxa da Perna/diagnóstico por imagemRESUMO
PURPOSE: The correction of severe post-tubercular kyphosis (PTK) is complex and has the disadvantage of being multistaged with a high morbidity. In this study, we evaluated the outcomes of children who underwent single-stage closing-opening wedge osteotomy as a surgical treatment of PTK of the thoracolumbar spine. METHOD: Our study group included 12 children with thoracolumbar PTK (seven boys and five girls) with an average age of 9.4 years (range 6-12 years), who were treated at our institution from January 2004 to October 2009. The American Spinal Injury Association Impairment Scale and visual analog scale score were used to classify neurologic function. All patients underwent halo-pelvic traction before surgery and were treated with single-stage closing-opening wedge osteotomy. RESULT: The duration of surgery averaged 99 min (range 70-150 min). Average blood loss was 782 ml (range 560-1,200 ml), and the average length of hospital stay was 12 days (range 8-16 days). The neurological function of all patients improved significantly after the procedure. The mean preoperative kyphotic angle was 83.3° (range 59-118°), which had reduced to 27.6° (range 20-38°) at the final follow-up visit. All patients had solid fusion, and no major complications were observed through the final follow-up visit. CONCLUSION: Single-stage closing-opening wedge osteotomy is an effective method to correct severe thoracolumbar PTK. A main advantage of the procedure is that it is a posterior-only, single-staged surgery, allowing for significant correction with minimal complications.
Assuntos
Cifose/cirurgia , Osteotomia/métodos , Complicações Pós-Operatórias/cirurgia , Fusão Vertebral/métodos , Tuberculose da Coluna Vertebral/cirurgia , Criança , Avaliação da Deficiência , Feminino , Humanos , Cifose/complicações , Estudos Longitudinais , Vértebras Lombares/cirurgia , Masculino , Vértebras Torácicas/cirurgia , Resultado do Tratamento , Tuberculose da Coluna Vertebral/complicações , Escala Visual AnalógicaRESUMO
PURPOSE: The aim of this study was to compare single posterior debridement, interbody fusion and instrumentation with one-stage anterior debridement, interbody fusion and posterior instrumentation for treating thoracic and lumbar spinal tuberculosis. METHOD: From January 2006 to January 2010, we enrolled 115 spinal tuberculosis patients with obvious surgical indications. Overall, 55 patients had vertebral body destruction, accompanied by a flow injection abscess or a unilateral abscess volume greater than 500 ml. The patients underwent one-staged anterior debridement, bone grafting and posterior instrumentation (group A) or single posterior debridement, bone grafting and instrumentation (group B). Clinical and radiographic results for the two groups were analyzed and compared. RESULTS: Patients were followed 12-36 months (mean 21.3 months), Fusion occurred at 4-12 months (mean 7.8 months). There were significant differences between groups regarding the post-operative kyphosis angle, angle correction and angle correction rate, especially if pathology is present in thoracolumbar and lumbar regions. Operative complications affected five patients in group A, and one patient in group B. A unilateral psoas abscess was observed in three patients 12 months postoperatively. In one of them, interbody fusion did not occur, and there was fixation loosening and interbody absorption. All of them were cured by an anterior operation. CONCLUSION: Anterior debridement and bone grafting with posterior instrumentation may not be the best choice for treating patients with spinal tuberculosis. Single posterior debridement/bone grafting/instrumentation for single-segment of thoracic or lumbar spine tuberculosis produced good clinical results, except in patients who had a psoas abscess.