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Phenylalanine ammonia lyase (PAL) can catalyze L-phenylalanine to produce trans-cinnamic acid, which is widely used in the fields of pharmacy, food and agriculture. In particular, phenylalanine ammonia lyase from Anabaena variabilis (AvPAL) is the only protein drug for the treatment of phenylketonuria. However, the poor activity and low stability limit the application in industry of AvPAL. In this study, the key amino acids of substrate-binding cavity in AvPAL were identified by screening the single site saturation mutagenesis library. Subsequently, the impact of replacing M222 with the additional 19 amino acids on activity was also evaluated by site-directed mutagenesis. It was found that the kcat values of mutants M222L and M222V were 90% and 60% higher than that of AvPAL, and the kcat/Km was 1.4 and 1.5 times as that of AvPAL. Molecular docking results revealed that the higher activity of M222L and M222V may be due to the increase of hydrophobicity favorable for the substrate-binding cavity. This study is important for elucidating the structure-function relationship of AvPAL.
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This present study aims to establish a UPLC method for simultaneously determining eleven components such as new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,rutin,hibisin and loganin in Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica and comparing the differences in the contents of phenolic acids,flavonoids and iridoid glycosides of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The method was carried out on an ACQUITY UPLC BEH C18column(2.1 mm×100 mm,1.7 μm) by a gradient elution using acetonitrile and 0.1% phosphoric acid.The flow rate was 0.3 mL·min-1.The column temperature was maintained at 30 ℃.The sample room temperature was 8 ℃.The wavelength was set at 326 nm for new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C,352 nm for rutin and lignin,and 238 nm for loganin.The injection volume was 1 μL.The eleven components has good resolution and was separated to baseline.Each component had a wide linear range and a good linear relationship(r≥0.999 6),the average recovery rate(n=9) was 98.96%,100.7%,97.24%,97.06%,99.53%,96.78%,98.12%,95.20%,95.12%,100.2%,98.61%and with RSD was 2.5%,1.4%,1.9%,2.1%,1.7%,1.9%,1.6%,2.0%,1.4%,2.2%,2.0%,respectively.Based on the results of the content determination,the chemometric methods such as cluster analysis and principal component analysis were used to compare the Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The results showed that Lonicerae Japonicae Flos and leaves of Lonicera japonica were similar in the chemical constituents,but both showed chemical constituents difference compored to Lonicerae Japonicae Caulis.The established multi-component quantitative analysis method can provide a reference for the quality control of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Flavonoids , Flowers , Chemistry , Hydroxybenzoates , Iridoid Glycosides , Lonicera , Chemistry , Phytochemicals , Plant Leaves , Chemistry , Quality ControlABSTRACT
Nowadays,with the rapid development of science and technology of medicine,the detection rate of obstructivc azoospermia increased gradually,clinically not uncommon.Surgical treatment of obstructive azoospermia is also improved from traditional vasovasostomy and epididymovasostomy to microsurgical vasovasos-tomy and microsurgical va-soepididymostomy,and from 3-suture intussusception vasoepididymostomy to 2-suture intussusception vasoepididymostomy,then our modified reverse single needle anastomosis,and finally to the robot assisted anastomosis.Therefore,the anastomosis is more precise,the recanalization rate and the pregnancy rate have a revolutionary improvement.The robot era is coming.The article focuses on surgical treatment of obstructive azoospermia,comparison of the advantages and disadvantages of each operation,and prediction of the future direction of development.
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Objective To observe Rongxin Pills in the treatment of viral myocarditis in children (deficiency of both qi and yin and heart meridian stasis syndrome) and the effectiveness and clinical application of safety.Methods Viral myocarditis patients (280 cases,deficiency of both qi and yin and heart meridian stasis syndrome),according to 3:1 ratio as the test group (n =21 0) and control group (n =70).The test group took orally Rongxin pills each time 4.5～9 g,3 times daily;the control group oral coenzyme Q10 capsule each time 10 ～ 20 mg,twice daily.The course of treatment was 28 d.The experiment was carried out with the random and double blind method.The symptoms of myocarditis,integrated and electrocardiogram,echocardiography,myocardial enzymes,as well as the efficacy of traditional Chinese medicine and improvement of the effect of the disease were observed.Results The results of FAS (PPS) analysis showed that 28 d after treatment,the symptom score and mean of experimental group and control group were 5.975 (6.000) and 4.721 (4.788).The syndromes of the total effective rates were 91.62％ (90.59％) and 70.59％ (71.21％),curative effect the total effective rates were 90.14％ (92.08％) and 72.06％ (72.73％).The total effective rate of experimental group was higher than that of the control group,the difference was statistically significant.In this experiment,three cases of clinical adverse events were reported,which were not related to the experimental drug.It also not belongs to adverse drug reactions.Conclusion Rongxin Pill in the treatment of viral myocarditis in children (deficiency of both qi and yin and heart meridian stasis syndrome) is more effective than coenzyme Q 10 capsule,and there was no indication of higher risk of clinical application.
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BACKGROUND:The third molar extraction may lead to different degrees of soft tissue damage. Ultrasound and ultrashort waves belong to hyperthermia, which can increase tissue temperature and improve the therapeutic efficacy of soft tissue stretch. OBJECTIVE:To observe the effects of soft tissue stretch combined with ultrasound and ultrashort wave therapy for patients who were recently diagnosed with trismus and pain folowing third molar removal. METHODS:Sixteen patients with trismus and pain after third molar removal were randomly divided into two groups: patients in stretch group received soft tissue stretch with ultrasound and ultrashort wave therapy, while those in control group received only ultrasound and ultrashort wave therapy. Patients received the treatment five times per week, totaly for 2 weeks. RESULTS AND CONCLUSION:It was found that after treatment pain was significantly reduced at the temporomandibular joint, and the range of maximum active mouth opening increased significantly. The improvement in mouth opening and temporomandibular joint function was better in the stretch group than the control group. These findings indicate that ultrasound and ultrashort wave therapy can improve symptoms of trismus and pain folowing third molar removal, and the combination of ultrasound and ultrashort wave therapy and soft tissue stretch therapy can achieve more effective results.
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BACKGROUND:Cartilage injury is stil one of the clinical problems difficult to be treated completely so far. Recently, the discovery of synovial mesenchymal stem cells (SMSCs) has brought about the new hope to cartilage repair. OBJECTIVE:To explore the process concerning SMSCs-based therapy for cartilage repair in the past few years, such as the characteristics of SMSCs, culture conditions, preclinical and clinical studies, and then to summarize the literatures published in recent years. METHODS:A computed-based online search of PubMed and SpringerLink databases was performed using the key words of“synovial mesenchymal stem cells, cartilage repair”for literatures published from January 1993 to May 2013. RESULTS AND CONCLUSION:Final y, 37 articles were included. SMSCs have a greater proliferative capability, colony-forming potential and chondrogenic potential than other mesenchymal stem cells. The diseases such as osteoarthritis and rheumatoid arthritis can influence the characteristics of SMSCs. Numerous articles have aimed at the studies of cellculture in vitro and celltransplantation in vivo. However, the process of SMSCs therapy is mostly at its preliminary stage. Reports on its unique characteristics, optimal culture conditions and the high-quality clinical studies are stil largely lacking. In a word, though further studies are needed, SMSCs appear to be a promising cellsource for cartilage repair in the future.
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Objective To observe the activation patterns in surface electromyographs (sEMGs) of the lumbar dorsal and ventral muscles and the gluteus medius during asymmetrical trunk postures and while carrying loads with the trunk rotated.Methods sEMGs were recorded from ten healthy men as they stood with their feet in 5 positions-rotated clockwise 0°,45°,90°,135° and 180° with respect to the initial plane of the torso.With each foot position they stood in 3 postures:upright,in lateral flexion and in lateral extension.Each posture was recorded with and without carrying a load of 20 kg by the right hand.The EMG activities of the rectus abdominis (RA),obliquus externus abdominis (EO),erector spinae (ES),multifidus (LM) and gluteus medius (GM) on both sides were recorded.Trunk displacement was monitored by 8 cameras.The EMG data of each muscle were normalized with readings from a maximal voluntary contraction (MVC) of the muscle.Results With the feet at 0° and no load the muscle activities on both sides were 1％-2％ of the MVC readings.With a 20 kg load the percentage was 3％-13％.With the feet at 45° and no load the activities were greater,except in the RA on the contralateral side.With no load at 90°,135° and 180° the muscle activities were greater,but a few muscles in the two extreme trunk postures showed greater activity only on one side,especially the ES and LM.With a load and rotational torque in the spine,the ventral muscles were less active but the dorsal muscles were significantly more active on both sides.Conclusion Standing with rotation of the spine,the activity of some trunk muscles shows great asymmetry,particularly with asymmetric loading of the arms.This might create a phenomenon of lumbar flashing and it could possibly be one of the pathological bases of acute lumbar sprain.
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Objective To observe electromyography (EMG) activity and kinematic characteristics of the dorsal and ventral muscles during extension-flexion of the trunk． Methods Ten healthy male subjects stood on a test platform, performing the front and back lifting and lowering. The EMG activity of 10 muscles (rectus abdominus, external oblique, gluteus medius, erector spinae, multifidus), the angular movement, the plantar center of pressure (COP) were recorded at the same time. The average amplitude of EMG, contribution ratio of muscles, and COP displacement were calculated, and statistical analyses of all the data were conducted． Results During front lifting and lowering, the activity of dorsal muscles was greater than that of ventral muscles under both loading and unloading, and the contribution ratio of dorsal muscles was increasing with loading. During back lifting and lowering, the activity of ventral muscles was slightly greater, but the contribution ratio of both dorsal and ventral muscles was very close. The contribution ratio of ventral muscles was increasing with loading, while the activity of dorsal muscles decreased. The activity of gluteus medius was increasing with loading, but the contribution ratio was basically consistent under both loading and unloading. Conclusions During extension-flexion of the trunk, muscle activation varies with the load size and trunk position. Under loading at front, dorsal muscles are working as an agonist, while ventral muscles as antagonist; under loading at back, ventral muscles become an agonist and dorsal muscles as a co-activator. The activity of gluteus medius might play a role of stabilization during front and back lifting and lowering.
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Objective To observe the electromyography (EMG) activities and kinematic characteristics of the back, abdomen and hip muscles under the static symmetric loading and unloading with the flexion and extension posture. Methods Six healthy male subjects stood on a specially designed testing platform, doing flexion/extension under symmetric loading and unloading, respectively. Each trial lasted 4 seconds and was repeated 3 times. The EMG activities of 10 muscles(rectus abdominis，obliquus externus abdominis，erector spinae，multifidus，gluteus Medius), the 3D angular movement, the plantar COP (center of pressure) were recorded during the trial.The normalized EMG, trunk angle and COP displacement were calculated, and statistical analyses of all data were made. Results Muscle activities of the back were greater (10.47~16.94) with flexion both under loading and unloading. Muscle activities of the abdomen were greater with extension under unloading, and those of the back (3.70~17.95) and hip (6.64~11.52) were increasing under loading, with muscle activities of the abdomen decreasing (10.66~4.18). The flashing trunk numbers was increased with the increase of loading, especially 1.55-time increase in the trunk angle. The COP displacement in anterior posterior direction was shifted greater than that in lateral direction, and this shift was more during flexion (14.60) than that during extension (7.65).Conclusions Extension increases the activities of back muscles, especially for multifidus, and it also increases the torso angular displacement and flashing trunk numbers, especially under loading.