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The last essential enzyme in the biosynthetic pathway of trilobatin, phloretin-4'-O glycosyltransferase (P4'-OGT), catalyzes the conversion of trilobatin to phloretin in vitro. However, only a few P4'-OGTs have been found in plants. This study used Malus domestica phloretin-4'-O glycosyltransferase (MdPh-4'-OGT) as a query to identify and clone two UDP-glucuronosyltransferase (UGT) genes, designated UGT74L2 and UGT74L3, from the transcriptome of Andrographis paniculata. According to a phylogenetic tree analysis, UGT74L2 and UGT74L3 belonged to the UGT74 family, which has been linked to several activities in other species. The in vitro enzymatic reaction demonstrated that UGT74L2 could particularly catalyze the formation of trilobatin from phloretin, but UGT74L3 had no effects. By using Ni-NTA affinity chromatography to extract the soluble UGT74L2 recombinant protein, the enzymatic kinetics of the activity was investigated using phloretin as the substrate. The results showed that the optimal temperature and pH for UGT74L2 enzymatic reaction were 40 ℃ and 8.0 (Tris-HCl system), respectively. Three metal ions (Ca2+, Mn2+ and Co2+) showed inhibitory effect on the activity of UGT74L2, while Mg2+ could improve the activity of UGT74L2. Other tested metal ions have no significant effect on UGT74L2. The results of enzymatic kinetic parameters that the Km value was 29.84 μmol·L-1, the kcat was 0.02 s-1, and the kcat·Km-1 was 572.6 mol-1·s-1. By homology modeling, molecular docking and mutation experiments, we found that multiple amino acids residues around the substrate binding pocket play quite an important role during catalytic process, In summary, we identified a novel P4'-OGT gene from medicinal plant Andrographis paniculata and provided a new efficient catalyst to synthesize trilobatin. Meanwhile, this study provides a reference for mining new efficient glycosylation modules from plants.
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Traditional Chinese medicine(TCM) carries the experience and theoretical knowledge of the ancients, and the use of "toxic" Chinese medicines is a major feature and advantage of TCM. "Toxic" Chinese medicines have unique clinical value and certain medication risk under the guidance of TCM theories such as compatibility for detoxification and treatment based on syndrome differentiation. In recent years, the safety events of Chinese medicines have occurred frequently, which has made the safety of Chinese medicine a public concern in China and abroad. However, limited by conventional cognitive laws and technical methods, basic research on toxicity of Chinese medicines fails to be combined with the clinical application. As a result, it is difficult to identify the clinical characteristics of, predict toxic and side effects of, or form a universal precise medication regimen for "toxic" Chinese medicines, which restricts the clinical application of them. In view of the problem that the toxicity of "toxic" Chinese medicines is difficult to be predicted and restricts the clinical application, the evidence-based research concept will provide new ideas for safe applcation of them in clinical practice. The integrated development of multiple disciplines and techniques in the field of big data and artificial intelligence will also promote the renewal and development of the research models for "toxic" Chinese medicines. Our team tried to propose the academic concept of evidence-based Chinese medicine toxicology and establish the data-intelligence research mode for "toxic" Chinese medicines and the intelligent risk prediction method for medicinal combination in the early stage, which provided methodological supports for solving the above problem. Thus, on the basis of summarizing the research status and problems of the clinical medication regimen of "toxic" Chinese medicines, our team took the evidence-based toxicology of TCM as the core concept, and tried to construct the multiple-evidence integrated evaluation and prediction method for "toxic" Chinese medicine, so as to guide the establishment of the non-toxic medication regimen of "toxic" Chinese medicines. Specifically, through the analysis of multivariate data obtained from the basic research, the evidence-based toxicology database of Chinese medicines and the individualized "toxicity-effect" intelligent prediction platform were built based on the disease-syndrome virtual patients, so as to identify the clinical characteristics and risks of "toxic" Chinese medicines and develop individualized medication regime. This study is expected to provide a methodological reference for the establishment of medication regimen and risk prevention strategy for "toxic" Chinese medicines. The method established in this study will bridge clinical research and basic research, enhance the transformation of the scientific connotation of attenuated compatibility, promote the development of evidence-based Chinese medicine toxicology, and ensure the clinical safety of "toxic" Chinese medicines.
Subject(s)
Artificial Intelligence , China , Drugs, Chinese Herbal/pharmacology , Humans , Medicine, Chinese Traditional , Research Design , SyndromeABSTRACT
Sweat gland is one of the important appendage organs of the skin, which plays an important role in thermoregulation and homeostasis maintenance. Sweat glands are damaged and unable to self-repair after burns, resulting in perspiration disorders eventually. However, current clinical strategies cannot restore the function of the damaged sweat glands effectively. Therefore, it is urgent to seek treatments that can promote the regeneration of sweat glands and restore their normal functions. Stem cells have extensive sources, low immunogenicity, high proliferation capacity, and multi-directional differentiation potential, which have become a focus in the field of regenerative medicine. In recent years, a variety of stem cells have been induced to differentiate into sweat gland-like tissue with certain secretory function, which provides treatment direction for sweat gland regeneration after burns in clinic. This article reviews the recent research advances on the application of stem cells in sweat gland regeneration from the perspectives of the manner by which stem cells transform into sweat gland cells in different environments and their influencing factors.
Subject(s)
Cell Differentiation/physiology , Regeneration/physiology , Skin , Stem Cells , Sweat Glands/physiologyABSTRACT
Objective:To study the relationship between morphologic classification and prognosis of patients with intrahepatic cholangiocarcinoma (ICC).Methods:The clinicopathologic data of 125 ICC patients who underwent R 0 resection at the Ningbo Medical Center Li Huili Hospital from January 2011 to May 2019 were retrospectively analyzed. This cohort consisted of 67 males and 58 females, aged 26.0 to 82.0 (63.5±9.5) years old. Based on the resected specimens, the patients were divided into the mass forming type, periductal infiltrating type and mixed type. The survival rates and recurrence-free survival outcomes of the patients among the three types were compared, and the risk factors that influenced prognosis were analyzed. Results:Using the predetermined inclusion and exclusion criteria, 16 patients with missing data and other causes of death were excluded from the initial 125 patients. Also, as the number of the mixed type was small, these patients were excluded. Finally 62 patients in the mass forming type and 42 patients in the periductal infiltrating type were analyzed. In the mass forming group, there were 32 males and 30 females, aged (63.1±9.1) years. In the perivascular infiltration group, there were 22 males and 20 females, aged (64.1±10.2) years. The 1-, 3-, 5-year survival rates of the mass forming group were 78.1%, 33.2%, 18.9% (median survival 25 months). The 1-, 3-, 5-year survival rates of the periductal infiltrating type were 63.3%, 8.3%, 6.1% (median survival 15 months). There were statistically significant differences between the two groups ( P<0.05). The median recurrence-free survival for the mass forming and the periductal infiltrating groups were 18 and 11 months, respectively, with a statistically significant differences between them ( P<0.05). Univariate and multivariate analysis showed that TNM staging Ⅲ~Ⅳ ( HR=2.966, 95% CI: 1.549-5.679) and periductal infiltrating type ( HR=2.403, 95% CI: 1.236-4.670) were independent risk factors for survival of these patients after operations. TNM staging Ⅲ~Ⅳ ( HR=2.466, 95% CI: 1.325-4.589), low grade differentiation ( HR=0.528, 95% CI: 0.299-0.934) and periductal infiltrating type ( HR=2.432, 95% CI: 1.295-4.565) were independent risk factors for relapse-free survival of these patients ( P<0.05). Conclusions:Morphological classification was found to be an independent risk factor for prognosis of ICC patients in this study, with significantly worse long-term prognosis when compared with the mass forming type.
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Objective@#To explore the predictive significance of logarithm of lymph node positive number to lymph node negative number (LODDS) in patients with gallbladder cancer, and to establish a predictive model of gallbladder cancer based on LODDS.@*Methods@#The clinical data of 1321 gallbladder cancer patients who underwent lymphadenectomy in SEER database from 2010 to 2014 were collected. They were randomly divided into test group (n=793) and verification group (n=528). COX multivariate analysis was used to determine independent prognostic factor, and a Nomogram prediction model was established. C index was used to evaluate the accuracy of patient Nomogram and TNM. ROC curve was used to evaluate the accuracy of the contour map and TNM staging in predicting survival rates.@*Results@#Independent risk factors for COX analysis were selected including age, tumor size, grade, TNM stage and LODDS analysis, and the contour map was drawn. In the experimental group, the C index was 0.744 (95% CI, 0.707-0.781), and the AUC values of ROC curves predicting 3-year and 5-year survival were 0.815 and 0.823, respectively. The predictive value of C index for TNM staging in the test group was 0.699 (95% CI, 0.662-0.736), and the AUC value of ROC curve for TNM staging predicting 3-year and 5-year survival was 0.779 and 0.790, respectively.@*Conclusion@#LODDS is an independent prognostic factor for gallbladder cancer. The prediction model based on LODDS is more accurate than TNM staging.
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Objective To explore the predictive significance of logarithm of lymph node positive number to lymph node negative number (LODDS) in patients with gallbladder cancer,and to establish a predictive model of gallbladder cancer based on LODDS.Methods The clinical data of 1321 gallbladder cancer patients who underwent lymphadenectomy in SEER database from 2010 to 2014 were collected.They were randomly divided into test group (n =793) and verification group (n =528).COX multivariate analysis was used to determine independent prognostic factor,and a Nomogram prediction model was established.C index was used to evaluate the accuracy of patient Nomogram and TNM.ROC curve was used to evaluate the accuracy of the contour map and TNM staging in predicting survival rates.Results Independent risk factors for COX analysis were selected including age,tumor size,grade,TNM stage and LODDS analysis,and the contour map was drawn.In the experimental group,the C index was 0.744 (95％ CI,0.707-0.781),and the AUC values of ROC curves predicting 3-year and 5-year survival were 0.815 and 0.823,respectively.The predictive value of C index for TNM staging in the test group was 0.699 (95 ％ CI,0.662-0.736),and the AUC value of ROC curve for TNM staging predicting 3-year and 5-year survival was 0.779 and 0.790,respectively.Conclusion LODDS is an independent prognostic factor for gallbladder cancer.The prediction model based on LODDS is more accurate than TNM staging.
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In review of recent literature on the treatment of heart failure with coronary heart disease by injection of Yiqi Fumai (freeze-dried), clinical evidence for the treatment of coronary heart disease with heart failure was reviewed from relevant systematic reviews, clinical randomized controlled trials, and safety evaluation, from the experiments in vivo and in vitro on its working mechanism of the treatment of coronary heart disease, heart failure, for more in-depth research on clinical and basic research in the future to lay the foundation.
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Traditional Chinese medicine (TCM) emphasizes treatment timing, and it believes that the timing of treatment is closely related to the curative effect. In the era of the Chinese and Western medicine combined application, it attaches new connotations on how to choose the timing of interventions. Defining the timing of intervention is one of the key links to make the best effect of Chinese medicine, and it is also the focus of clinical decision-making, which deserves research attention. This article expounded the significance of TCM intervention timing research, and pointed out that the alternative timing should be based on the baseline investigation and mechanism clues. This article also demonstrated some key points of intervention timing trial design, like the outcome indicators setting, sample size estimation, blinding and data analysis. With the examples of national key research and development program"evidence-based optimization and mechanism research of traditional Chinese medicine therapies for coronary artery disease (angina pectoris-myocardial infarction-heart failure) ", the timing of TCM intervention in myocardial infarction perioperative period and coronary heart disease related research cases aboard, we hope to provide reference for the intervention timing optimization research.
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Objective: To conduct a comprehensive and systematic review of the efficacy and safety of Wenxin Keli (WXKL) in the treatment of atrial fibrillation (AF) . Methods: Seven databases (PubMed, The Cochrane Library, Web ofScience, CNKI, Wanfang Database, VIP and SinoMed) were searched to identify relevant randomized controlled trials (RCTs) from inceptions to 1 October, 2018. Two review authors independently assessed the methodological quality andanalyzed data by Cochrane handbook and the Rev Man 5.3 software. Begg.s test was conducted to assess publication biasvia Stata 14 software. Results: Twenty-four RCTs with 2246 patients were included in this review. Compared with blankcontrol, placebo or western medicine alone, WXKL alone or combined with western medicine could effectively reducerapid ventricular rate (MD=-7.14, 95％CI:-8.42——5.87), the frequency and duration of AF. It could also shorten thesinus rhythm conversion time (MD=-3.04, 95％CI:-3.47——2.61), increase the sinus rhythm conversion rate (RR=1.19, 95％ CI: 1.09～1.29) and decrease recurrence rate of AF (RR=0.28, 95％ CI: 0.13-0.59) . Besides, WXKL alone orcombined with western medicine was beneficial for improving the left ventricular ejection fraction (LVEF) (MD=3.44, 95％ CI: 0.87-6.01), left ventricular end diastolic diameter (LVEDD) (MD=-2.47, 95％ CI:-2.86——2.08), left atrialdiameter (LAd) (MD=-0.91, 95％CI:-1.58——0.25) and P wave dispersion (Pd) (MD=-4.04, 95％CI:-4.15——3.93) .WXKL combined with low-dose amiodarone was superior to conventional-dose amiodarone alone in improving themaximum P wave (Pmax) (MD=-8.25, 95％ CI:-10.33——6.17), and WXKL combined with conventional-doseamiodarone is more effective (MD=-13.10, 95％CI:-13.65——12.55) . Compared with the control group, the treatmentgroup had fewer adverse reactions, and the Begg.s test did not find any publication bias. Conclusion: WXKL alone orcombined with western medicine exhibited better therapeutic effects in the treatment of AF, but these results still needhigh-quality evidence to verify.
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Background Corneal collagen cross-linking (CXL) shows good clinical effects for keratoconus,and de-epithelized CXL appears to be benefit to the distribution and absorption of riboflavin in cornea stroma.However,de-epithelization of CXL will increase the infective risk and corneal healing time.It is very important to understand and control the affecting factors of corneal repair after de-epithelization of CXL.Objective This study was to evaluate the characteristics of corneal epithelial repair and analyze the relevant factors affecting corneal healing time after de-epithelized CXL.Methods A series-cases observational study was performed.De-epithelized CXL was performed on 77 eyes of 68 keratoconus patients in Shandong Eye Hospital from September 2013 to September 2015 under the approval of Ethic Committee of this hospital and informed consent of each patient.The age,corneal curvature,corneal thickness,breakup time of tear film (BUT),corneal front astigmatism (Astig) and epithelial healing time of the patients were recorded after surgery.The correlations between corneal epithelium healing time and above-mentioned factors were analyzed.Results De-epithelized CXL was smoothly finished in all the eyes.The corneal epithelium healing time was 2-12 days after surgery,with the average healing time 5 (4,6) days.The mean age,thickness at corneal thinnest point,minimal cornea curvature (Kf),maximal corneal curvature (Ks),corneal average curvature (Km) and Astig was 22.00 (18.00,25.00) years,436 (412,470) μm,47.40 (44.70,50.45) D,52.10 (49.00,54.55) D,50.00 (47.15,53.15) D and-3.30 (-5.45,1.70) D,respectively.Spearman rank correlation analysis showed significant negative correlations between corneal epithelium healing time and BUT or the thickness at corneal thinnest point (BUT:rs =-0.334,P =0.003;corneal thickness:rs =-0.417,P =0.000),and thesignificant positive correlations were found between corneal epithelium healing time and Km,Kf and Ks (Km:rs =0.449,P =0.000;Kf:rs =0.300,P =0.008;Ks:rs =0.432,P =0.000).There were no considerable correlarions between corneal epithelium healing time and age or Astig (age:rs =0.023,P =0.845;Astig:rs =-0.190,P =0.098).Multiple linear regression analysis were carried out to study the dependent variable and independent factors.Because of the multiple co-linearity between variables,this paper corrects the model by using ridge regression.There is significant negative correlation between BUT,corneal thickness and corneal healing time,respectively (both at P＜0.05),corneal curvature Km and Kf is positively correlated with corneal healing time (both at P ＜ 0.05).Conclusions The corneal thickness,Kf,Km,as well as BUT are influencing factors of epithelial healing after CXL.
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Objective To analyze the related risk factors for delayed gastric emptying ( DGE) after pancreaticoduodenectomy .Methods Clinical data on 308 patients who underwent pancreaticoduodenectomy at Ningbo Lihuili hospital from January 2009 to December 2014 were retrospectively analyzed , and patients were divided into DGE group and non-DGE group.Univariate analysis and multivariate logistic regression analysis were used to study the risk factors associated with DGE during perioperative period .Results DGE occurred in 55 patients (17.9%).The incidences of grade A, grade B and grade C DGE were 7.1%(22/308), 6.2%(19/308) and 4.5%(14/308), respectively.The univariate analysis showed the method of pancreatic digestive tract reconstruction ( pancreaticogastrostomy or pancreaticojejunostomy ) , postoperative pancreatic fistula, postoperative biliary fistula and postoperative intraabdominal infection were risk factors for DGE after surgery. Multivariate analysis indicated that the method of pancreatic digestive tract reconstruction (OR=1.19, P=0.046), postoperative pancreatic fistula ( OR=1.33, P=0.014), postoperative biliary fistula (OR=1.43, P=0.047) and postoperative intraabdominal infection (OR=1.51, P=0.001) were independently associated with DGE . Postoperative pancreatic fistula (OR=3.692, P=0.021) and intraabdominal infection (OR=3.725,P=0.003)were also the independent risk factors for Grade B and Grade C DGE. Conclusions DGE after pancreaticoduodenectomy was strongly related to the postoperative complications .Postoperative pancreatic fistula , biliary fistula and intraabdominal infection were associated with increased risk of DGE , while pancreaticogastrostomy reduced the incidence of DGE by decreasing the incidence of pancreatic or biliary fistula .
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<p><b>BACKGROUND</b>The optimal age at which to initiate for auricular reconstruction is controversial. Rib cartilage growth is closely related to age and determines the feasibility and outcomes of auricular reconstruction. We developed a method to guide the timing of auricular reconstruction in children with microtia ranging in age from 5 to 10 years.</p><p><b>METHODS</b>Rib cartilage and the healthy ear were assessed using low-dose multi-slice computed tomography. The lengths of the eighth rib cartilage and the helix of the healthy ear (from the helical crus to the joint of the helix and the earlobe) were measured. Surgery was performed when the two lengths were approximately equal.</p><p><b>RESULTS</b>The preoperative eighth rib measurements significantly correlated with the intraoperative measurements (P < 0.05). From 5 to 10 years of age, eighth rib growth was not linear. In 76 (62.8%) of 121 patients, the eighth rib length was approximately equal to the helix length in the healthy ear; satisfactory outcomes were achieved in these patients. In 18 (14.9%) patients, the eighth rib was slightly shorter than the helix, helix fabrication was accomplished by adjusting the length of the helical crus of stent, and satisfactory outcomes were also achieved. Acceptable outcomes were achieved in 17 (14.0%) patients in whom helix fabrication was accomplished by cartilage splicing. In 9 (7.4%) patients with insufficient rib cartilage length, the operation was delayed. In one (0.8%) patient with insufficient rib cartilage length, which left no cartilage for helix splicing, the result was unsatisfactory.</p><p><b>CONCLUSIONS</b>Eighth rib cartilage growth is variable. Rib cartilage assessment relative to the healthy ear can guide auricular reconstruction and personalize treatment in young patients with microtia.</p>
Subject(s)
Age Factors , Autografts , Cartilage , Diagnostic Imaging , Transplantation , Child , Child, Preschool , Congenital Microtia , General Surgery , Ear, External , Diagnostic Imaging , Female , Humans , Male , Plastic Surgery Procedures , Ribs , Diagnostic Imaging , Tomography, X-Ray ComputedABSTRACT
ECM is a supporting structure for stabilizing the location of cells and preserving the structure of tissues. Recently, it has been discovered that ECM and its degradation products may exert profound influences on tissues and cells, such as activities of inflammatory cells and immune cells. Angiogenesis may be stimulated or inhibited by degradation products of ECM. Matrikines, liberated by partial proteolysis of ECM macromolecules, are found to regulate cell functional activities and play a significant role in wound healing or tumor invasion. Post-burn denatured dermal matrix is being studied in burn healing now. The study of post-burn denatured or necrotic dermal matrix should be emphasized in future.
Subject(s)
Animals , Extracellular Matrix , Metabolism , Humans , Inflammation , Metabolism , Wound HealingABSTRACT
Objective To identify the miR-217 targeted gene ANLN by experiment.Methods Bioinformatic algorithms were used to predict the potential targets of miR-217.Then,ANLN binding with miR217 and mutant ANLN (mutANLN) sequence were designed and synthesized,and their amplified fragments were inserted into plasmid psiCHECK-2,and recombinant plasmid psiCHECK-2-ANLN and psiCHECK-2-mutANLN were reconstructed.The two recombinant plasmids were co-transfected into pancreatic cancer cell line PANC1 with miR-217,miR-217 inhibitor,NC,NC inhibitor by liposome,respectively.Dual luciferase reporter system was used to determine the luciferase activity,and Western blot was used to measure the expression of ANLN protein.Results The luciferase activities of psiCHECK-2-ANLN,psiCHECK-2-ANLN +miR-217,psiCHECK-2ANLN + miR-217 inhibitor,psiCHECK-2ANLN + NC,psiCHECK-2-ANLN + NC inhibitor were 2.221 ± 0.188,0.769 ± 0.061,3.764 ± 0.371,2.265 ± 0.201,2.242 ± 0.018,and the difference among these groups was statistically significant (F =77.405,P ＜0.001),but the difference among psiCHECK-2ANLN group,psiCHECK-2-ANLN + NC group and psiCHECK-2-ANLN + NC inhibitor group was not statistically significant.However,luciferase activities of psiCHECK-2-ANLN + miR-217 group were significantly decreased when compared with other 3 groups,and luciferase activity of psiCHECK-2-ANLN +miR-217 inhibitor group were significantly increased when compared with other 4 groups (all P ＜0.001).Luciferase activities of groups transfected with psiCHECK-2-mutANLN was not significantly different (P =0.053).The expression of ANLN protein in PANC1 with psiCHECK-2-ANLN + miR-217 co-transfection was significantly down-regulated when compared with that with psiCHECK-2-ANLN transfection alone.Conclusions ANLN is one of the direct target genes of miR-217 in PANC1 cells.
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Objectives To investigate the prevalence of non-alcoholic fatty liver (NAFLD) disease and its correlation with chronic metabolic diseases in two primary school students in Shanghai. Methods One thousand ifve hundred and thirty-two 7-11 year-old students from two primary schools were enrolled in Septamber-October 2011. The anthropometric indices, blood pressure, screening for pseudoacanthosis nigricans and liver ultrasonography of all subjects were recorded. Results The overall prevalence of NAFLD, obesity, abdominal obesity, pseudoacanthosis nigricans, high systolic blood pressure and high diastolic blood pressure was 6.5%, 26.7%, 16.3%, 5.1%, 1.7%and 1.9%, respectively. The prevalence of NAFLD and abdomi-nal obesity in students of central urban area was signiifcantly higher than that in suburban area (P<0.01). The binary regression analysis revealed a signiifcant association between NAFLD in students with sex, age, obesity, abdominal obesity, pseudoacan-thosis nigricans and high systolic blood pressure (P<0.05). Conclusions NAFLD has close correlation with chronic metabolic disease in children. It is time to adopt prevention, detection and treatment in NAFLD children with symptoms of chronic meta-bolic diseases.
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Sucrose phosphorylase (EC 2.4.1.7, Sucrose phosphorylase, SPase) can be produced by recombinant strain Escherichia coli Rosetta(DE3)/Pet-SPase. Crude enzyme was obtained from the cells by the high pressure disruption and centrifugation. Sucrose phosphorylase was purified by Ni-NTA affinity column chromatography and desalted by ultrafiltration. The specific enzyme activity was 1.1-fold higher than that of the crude enzyme, and recovery rate was 82.7%. The purified recombinant SPase had a band of 59 kDa on SDS-PAGE. Thermostability of the enzyme was shown at temperatures up to 37 degrees C, and pH stability between pH 6.0 and 6.7. The optimum temperature and pH were 37 degrees C and 6.7, respectively. The K(m) of SPase for sucrose was 7.3 mmol/L, and Vmax was 0.2 micromol/(min x mg). Besides, alpha-arbutin was synthesized from sucrose and hydroquinone by transglucosylation with recombinant SPase. The optimal conditions for synthesis of alpha-arbutin were 200 U/mL of recombinant SPase, 20% of sucrose, and 1.6% hydroquinone at pH 6-6.5 and 25 degrees C for 21 h. Under these conditions, alpha-arbutin was obtained with a 78.3% molar yield with respect to hydroquinone, and the concentration of alpha-arbutin was about 31 g/L.
Subject(s)
Arbutin , Catalysis , Enzyme Stability , Escherichia coli , Genetics , Glucosyltransferases , Genetics , Metabolism , Hydroquinones , Metabolism , Recombinant Proteins , Genetics , Metabolism , Sucrose , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the feasibility of burn denatured acellular dermal matrix (DADM) as dermal substitute in repairing wounds.</p><p><b>METHODS</b>(1) Nine Wistar rats received a deep partial-thickness scald on the back. Full-thickness wounded skin was collected on post scald day (PBD) 1, 2, and 3 (with 3 rats at each time point), and it was treated with 2.5 g/L trypsin/0.5% Triton X-100 to remove cells to prepare DADM, respectively called DADM-1 d, DADM-2 d, and DADM-3 d. Another 3 rats without scald injury were treated with the same method as above to prepare acellular dermal matrix (ADM) to serve as control. Gross and histological observations and microbiological and biomechanical tests, including ultimate tensile strength, maximum tension, stretched length at breaking, stress-strain relationship, were conducted for the resulting ADM and DADM. (2) Another 64 rats were divided into ADM group and DADM-1 d, DADM-2 d, and DADM-3 d groups according to the random number table, with 16 rats in each group. A skin flap in size of 2.0 cm×1.8 cm was raised on the back of each rat. The above-mentioned ADM, DADM-1 d, DADM-2 d, and DADM-3 d were cut into pieces in the size of 1.8 cm×1.5 cm, and they were respectively implanted under the skin flaps of rats in corresponding group. At post surgery week (PSW) 1, 3, 5, or 9, 4 rats in each group were used to observe wound healing condition and change in implants with naked eye, and histological observation of the implants was conducted. Data were processed with one-way analysis of variance and t test.</p><p><b>RESULTS</b>(1) The freshly prepared DADM was milky white, soft in texture with flexibility, but poor in elasticity as compared with ADM. No epithelial structure or cellular component was observed in ADM or DADM under light microscope. Collagen fibers of DADM were seen to be thickened unevenly and arranged in disorder and eosinophilic. All microbiological results of DADM were negative. There was no statistically significant difference among DADM-1 d, DADM-2 d, and DADM-3 d in levels of ultimate tensile strength, maximum tension, stretched length at breaking, and stress-strain relationship (with F values from 0.088 to 3.591, P values all above 0.05). Values of the above-mentioned four indexes were the highest in DADM-3 d, they were respectively (13.0 ± 2.4) MPa, (61 ± 4) N, (173 ± 7)%, (45.7 ± 2.0)%. Values of the four indexes of ADM were respectively (19.0 ± 2.6) MPa, (95 ± 4) N, (201 ± 5)%, (62.5 ± 2.2)%, which were higher than those of DADM-1 d, DADM-2 d, and DADM-3 d (with t values from 6.424 to 17.125, P values all below 0.01). (2) No exudate or swelling in the wounds of rats, and no contraction or curling of implants were observed in every group at PSW 1, but inflammatory cells infiltration and Fbs inward migration were observed in the wound. At PSW 3, the growth of hair was normal in the wound in DADM-1 d, DADM-2 d, and ADM groups, but few and scattered hair grew in DADM-3 d group. The inflammatory cells decreased, while Fbs increased, and new capillaries were found to grow inwardly in each group. The decrease in inflammatory cells was slightly delayed in DADM-3 d group. At PSW 5, hair growth became normal, and implants shrank and thinned with fiber membrane wrapped densely and bundles of ingrowing large caliber blood vessels in all groups. The dermal matrix in each group merged with the surrounding normal tissue. At PSW 9, ADM and DADM became white, thin, and soft tissue sheet which was closely connected with the inner side of the flap. There was no infiltration of inflammatory cells in implants in either group. The collagen fibers arranged regularly and densely, and they were integrated with normal collagen tissue.</p><p><b>CONCLUSIONS</b>The burned DADM does not have obvious immunogenicity, but with good biocompatibility. It is prospective to become as a dermal substitute in repairing wounds.</p>
Subject(s)
Acellular Dermis , Animals , Burns , Pathology , General Surgery , Female , Male , Rats , Rats, Wistar , Plastic Surgery Procedures , Methods , Skin , Wounds and Injuries , Skin Transplantation , Methods , Skin, Artificial , Wound HealingABSTRACT
The aim of this study is to investigate the feasibility of silica-coated ethosome as a novel oral delivery system for the poorly water-soluble curcumin (as a model drug). The silica-coated ethosomes loading curcumin (CU-SE) were prepared by alcohol injection method with homogenization, followed by the precipitation of silica by sol-gel process. The physical and chemical features of CU-SEs, and curcumin release were determined in vitro. The pharmacodynamics and bioavailability measurements were sequentially performed. The mean diameter of CU-SE was (478.5 +/- 80.3) nm and the polydispersity index was 0.285 +/- 0.042, while the mean value of apparent drug entrapment efficiency was 80.77%. In vitro assays demonstrated that CU-SEs were significantly stable with improved release properties when compared with curcumin-loaded ethosomes (CU-ETs) without silica-coatings. The bioavailability of CU-SEs and CU-ETs was 11.86- and 5.25-fold higher, respectively, than that of curcumin suspensions (CU-SUs) in in vivo assays. The silica coatings significantly promoted the stability of ethosomes and CU-SEs exhibited 2.26-fold increase in bioavailablity relative to CU-ETs, indicating that the silica-coated ethosomes might be a potential approach for oral delivery of poorly water-soluble drugs especially the active ingredients of traditional Chinese medicine with improved bioavailability.
Subject(s)
Administration, Oral , Animals , Biological Availability , Coated Materials, Biocompatible , Chemistry , Curcumin , Chemistry , Pharmacokinetics , Drug Carriers , Ethanol , Chemistry , Liposomes , Chemistry , Male , Particle Size , Rats , Rats, Sprague-Dawley , Silicon Dioxide , Chemistry , SolubilityABSTRACT
The aim of this study is to investigate the feasibility of silica-coated ethosome as a novel oral delivery system for the poorly water-soluble curcumin (as a model drug). The silica-coated ethosomes loading curcumin (CU-SE) were prepared by alcohol injection method with homogenization, followed by the precipitation of silica by sol-gel process. The physical and chemical features of CU-SEs, and curcumin release were determined in vitro. The pharmacodynamics and bioavailability measurements were sequentially performed. The mean diameter of CU-SE was (478.5 +/- 80.3) nm and the polydispersity index was 0.285 +/- 0.042, while the mean value of apparent drug entrapment efficiency was 80.77%. In vitro assays demonstrated that CU-SEs were significantly stable with improved release properties when compared with curcumin-loaded ethosomes (CU-ETs) without silica-coatings. The bioavailability of CU-SEs and CU-ETs was 11.86- and 5.25-fold higher, respectively, than that of curcumin suspensions (CU-SUs) in in vivo assays. The silica coatings significantly promoted the stability of ethosomes and CU-SEs exhibited 2.26-fold increase in bioavailablity relative to CU-ETs, indicating that the silica-coated ethosomes might be a potential approach for oral delivery of poorly water-soluble drugs especially the active ingredients of traditional Chinese medicine with improved bioavailability.
ABSTRACT
<p><b>BACKGROUND</b>Transforming growth factor-β1 (TGF-β1) is known to have a role in keloid formation through the activation of fibroblasts and the acceleration of collagen deposition. The objective of this current study was to isolate TGF-β1 phage model peptides from a phage display 7-mer peptide library to evaluate their therapeutic effect on inhibiting the activity of keloid fibroblasts.</p><p><b>METHODS</b>A phage display 7-mer peptide library was screened using monoclonal anti-human TGF-β1 as the target to obtain specific phages containing ectogenous model peptides similar to TGF-β1. Enzyme-linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity, which underwent DNA sequencing. MTT assay and apoptosis assessment were used to evaluate the biological effects of the phage model peptides on keloid fibroblasts. Immunofluorescence assay was employed to show the binding affinity of the model peptides on phages causing keloid fibroblasts. Quantitative real-time PCR analysis was carried out to detect the expressions of nuclear factor κB (NF-κB) mRNA, connective tissue growth factor (CTGF) mRNA and TGF-β receptor II (TβRII) mRNA in keloid fibroblasts.</p><p><b>RESULTS</b>Specific phages with good results of ELISA were beneficiated. Four phage model peptides were obtained. The data of MTT showed that TGF-β1 and one phage model peptide (No. 4) could promote keloid fibroblasts proliferation, however, three phage model peptides (No. 1 - 3) could inhibit keloid fibroblasts proliferation. The results of apoptosis assessment showed that the three phage model peptides could slightly induce the apoptosis in keloid fibroblasts. The data of immunofluorescence assay revealed that the model peptides on phages rather than phages could bind to keloid fibroblasts. The findings of quantitative real-time PCR analysis suggested that the expressions of NF-κB mRNA and CTGF mRNA in the three phage model peptide groups decreased, while the expression of TβRII mRNA slightly increased.</p><p><b>CONCLUSIONS</b>Three phage model peptides isolated from a phage display 7-mer peptide library can inhibit keloid fibroblasts proliferation and induce the apoptosis in keloid fibroblasts. They can inhibit the activity of keloid fibroblasts by blocking TGF-β1 binding to its receptor and then regulating the expressions of NF-κB, CTGF and TβRII.</p>