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Arabinose-5-phosphate isomerase (KdsD) is the first key limiting enzyme in the biosynthesis of 3-deoxy-D-manno-octulosonate (KDO). KdsD gene was cloned into prokaryotic expression vector pET-HTT by seamless DNA cloning method and the amount of soluble recombinant protein was expressed in a soluble form in E. coli BL21 (DE3) after induction of Isopropyl β-D-1-thiogalactopyranoside (IPTG). The target protein was separated and purified by Ni-NTA affinity chromatography and size exclusion chromatography, and its purity was more than 85%. Size exclusion chromatography showed that KdsD protein existed in three forms: polymers, dimmers, and monomers in water solution, different from microbial KdsD enzyme with the four polymers in water solution. Further, the purified protein was identified through Western blotting and MALDI-TOF MASS technology. The results of activity assay showed that the optimum pH and temperature of AtKdsD isomerase activities were 8.0 and 37 ℃, respectively. The enzyme was activated by metal protease inhibitor EDTA (5 mmol/L) and inhibited by some metal ions at lower concentration, especially with Co²⁺ and Cd²⁺ metal ion. Furthermore, when D-arabinose-5-phosphate (A5P) was used as substrate, Km and Vmax of AtKdsD values were 0.16 mmol/L, 0.18 mmol/L·min. The affinity of AtKdsD was higher than KdsD in E. coli combined with substrate. Above results have laid a foundation for the KdsD protein structure and function for its potential industrial application.
Subject(s)
Aldose-Ketose Isomerases , Arabidopsis , Arabidopsis Proteins , Cloning, Molecular , Escherichia coli , Metabolism , Metals , Pentosephosphates , Recombinant ProteinsABSTRACT
Objective To investigate the effect of integrated swallowing function rehabilitation training on nasal feeding patients with pseudobulbar palsy or bulbar palsy after acute ischemic stroke. Methods A total of 54 patients met the diagnosis criteria were enrolled prospectively. They were divided into either an integrated group (n=31)or a control group (n=23)using a random number table method. The patients of the control group received routine treatment of stroke,including brain protection therapy,improvement of cerebral circulation,dehydration,regulating blood pressure,and symptomatic treatment,etc;in addition to conventional treatment,those of the integrated group also received integrated swallowing function rehabilitation training,including neuromuscular electrical stimulation plus swallowing reflex facilitation technique,and oral facial glossopharyngeal function training,etc. The oral intake function in patients of the integration and pseudobulbar palsy or bulbar palsy (according to MRI and signs)was compared between the two groups. The evaluation index was the Functional Oral Intake Scale (FOIS)score. Results (1)The FOIS score (median 4)of the integrated group was higher than that of the control group (median 2). There was significant difference between the two groups (P<0. 01). There was also significant difference before and after comparison between the two groups. (2)The FOIS score of pseudobulbar palsy in the integrated group was higher than that of the control group (median scores 4 and 2 ). There was significant difference (P<0.01);There was no significant difference in the FOIS scores of bulbar palsy between the two groups,but the FOIS score of the integrated group was higher than that of the control group (median scores 4 and 3). (3)The median differences between the integrated group and the control group were 2 and 1 respectively. The increased amplitude of FOIS of the integrated group was higher than that of the control group. There was significant difference (P<0. 01). Conclusion The integrated swallowing function rehabilitation training can improve the oral intake function,significantly reduce irritating cough,increase the capacity of oral intake,and increase the types of food. The effect of the improvement of pseudobulbar palsy is more significant in the integrated group.
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@#Objective To investigate the effects of repetitive transcranial magnetic stimulation (rTMS) on motor recovery in patients with incomplete spinal cord injury （SCI）. Methods 22 patients with T5～T12 incomplete spinal cord injury (ASIA C) were randomly divided into rTMS group and control group. 12 ptients in rTMS group were treated with 10Hz rTMS （10×5 s with 2 min interval) on motor cortex and rehabilitation. 10 patients in control group treated with rehabilitation only. They were assessed with ASIA motor score, ASIA pin prick score, ASIA light touch score, range of motion (ROM) of legs in antigravitation position, walking index for spinal cord injury Ⅱ (WISCIⅡ) and functional independence measure (FIM) score before and 4 weeks after treatment. Results There was no difference between two groups with the clinical assessment before treatment （P>0.05）. The ASIA motor score, ROM of legs and FIM score improved in both groups after treatment （P<0.05）. The score of WISCIⅡ also improved in rTMS group （P<0.05）, and ASIA motor score improved more in rTMS group than that of control group （P<0.05）. Conclusion rTMS can facilitate the recovery of the motor and walking function in patient with incomplete spinal cord injury.
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@# Objective To explore the role of architecture of the human muscle in treatment of spasticity after stroke with BTX.Methods40 patients with spasticity hemiplegia after stroke were involved,whose scores of Modified Ashworth Scale beyond 3.They were divided in 2 groups:control group and observed group,in which BTX dose and injection points designed without or with architecture of the human muscle.All muscles injected were located with electrical stimulator.They were evaluated with Modified Ashworth Scale and the incidence of brace wearing were observed 2 weeks after treatment.ResultsThe Ashworth scores for wrist flexors were(1.3±0.9234)in observed group,(1.9±0.8522)in control group(P<0.05);for the triceps surae,they were(1.6±0.8826)and(2.25±0.7864)respectively(P<0.05).10 cases wore brace for upper limbs in observed group and 14 cases in control group;for the brace for lower limbs,there were 12 in observed group and 16 in control group(P>0.05).ConclusionBTX dose and injection points designed by Architecture of the Human Muscle in Treatment of spasticity after stroke is better than routine way,but the brace wearing for patients showed no significant improvement.
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OBJECTIVE:To compare the in vitro antibacterial activity of azlocillin given alone with azlocillin plus tazobactam against168strains of clinically isolated pathogenic bacteria.METHODS:The antibacterial activities of two antibac_ terials against168strains of clinically isolated pathogenic bacteria in vitro were detected by two-fold dilution method.RE-SULTS:The MIC 50 and the MIC 90 of the combined therapy of azlocillin/tazobactam(4∶1)were1/32and1/64,respectively that of azlocillin given alone.CONCLUSION:The concomitant therapy of azlocillin with tazobactam improves the antibacterial activity.