ABSTRACT
Glutamine synthetase is a key enzyme in plant nitrogen assimilation. To study the structure of wheat glutamine synthetase isoenzymes, GS1, GSr, GSe, GS2 and GS2p of wheat were cloned into pET-21a, and the expression condition was optimized. Although wheat glutamine synthetase isoenzymes had 70%-80% amino acid sequence homology, the isoforms expressed with different characteristics. Induced at 30 °C, the most expression level of GSr, GSe and GS2 was after 3 h, and of GS1 was at the 7 h whereas no GS2p was expressed, and the GS isoenzymes showed different expression level, with the order of GS1 (22%)>GSr (15%)>GS2 (12%)>GSe (5%). GSe expressed as soluble protein, and GS1 expressed mainly as soluble protein whereas GSr and GS2 expressed as insoluble proteins. Induced at 30 °C for 3 h, mRNA transcript levels of GS isoforms were different, with the order of GSr (7.59)>GS2 (1.84)>GS2p (1.66)>GSe (1.46)>GS1 (1.00). The levels of mRNA transcription were not consistent with the level of the protein translation. The analysis of mRNA secondary structure showed the free energy of translation initiation region of glutamine synthetase isoforms was different, with the order of GS1 (14.4)<GSr (17.2)<GS2 (22.6) <GSe (25.4) <GS2p (31.6), the smaller freed energy, the more unstable mRNA secondary structure of translation initiation region and the higher level of protein expression. Soluble expression condition of glutamine synthetase isozymes was also different, with GS1, GSr, GSe and GS2 induced at 30 °C for 5 h, 16 °C for 15 h, 37 °C for 5 h, and 25 °C for 7 h respectively. The soluble protein showed different expression level with GS1 (20%)>GSr (13%)>GS2 (10%)>GSe (7%), and different activities with GS1>GSe>GS2, and the activity of GSr was not detected. The gene sequence of glutamine synthetase isoenzymes determines the amount, status and activity of proteins expressed in prokaryotic cells.
ABSTRACT
Objective To investigate the curative effect and safety of percutaneous intradiscal injection of ozone combined with epidural injection of collagenase for the treatment of prominent lumbar disc protrusion.Methods A total of 541 patients with lumbar disc protrusion were included in this study.All the patients fulfilled the following criteria:the fibrous ring was broken,the protruded extent of nucleus pulposus was less than 10 mm and the dura sac and/or nerve root was compressed.The intradiscal injection of ozone combined with epidural injection of collagenase was performed in all patients.The patients were followed up through further consultation,letters,telephone or other ways.Five hundred and forty-one cases were followed up for 2-3 months(short-term),312 cases for 4-12 months(medium-term)and 115 cases for 13-18 months(ling-term).The short-term,medium-term and long-term curative effects were evaluated with unified curative criteria,and the results were compared with that obtained with the treatment of epidural injection of collagenase only. Results The short-term, medium-term and long-term effective rate of selected cases was 95.9%(519/541),90.4%(489/541)and 87.2%(472/541),respectively.When combined application of two methods was performed,different degree of shrinkage of the protruded nucleus pulposus was obviously observed,with a mean shrinkage degree of 30.5%.The short-term,medium-term and long-term effective rate of simple collagenase chemonucleolysis was 89.5%,82.4%and 80.4%,respectively.Conclusion Intradiscal injection of ozone combined with epidural injection of collagenase is an effective treatment for lumbar disc protrusion when the fibrous ring is broken,the protruded extent of nucleus pulposus is less than 10 mm and the dura sac and/or nerve root is compressed. Its curative effect is superior to simple collagenase chemonucleolysis. Ozone is of clinical significance for producing the shrinkage of the nucleus pulposus.
ABSTRACT
Objective To investigate the curative effect and safety of percutaneous intradiscal injection of ozone combined with epidural injection of collagenase for the treatment of prominent lumbar disc protrusion.Methods A total of 541 patients with lumbar disc protrusion were included in this study.All the patients fulfilled the following criteria:the fibrous ring was broken,the protruded extent of nucleus pulposus was less than 10 mm and the dura sac and/or nerve root was compressed.The intradiscal injection of ozone combined with epidural injection of collagenase was performed in all patients.The patients were followed up through further consultation,letters,telephone or other ways.Five hundred and forty-one cases were followed up for 2-3 months(short-term),312 cases for 4-12 months(medium-term)and 115 cases for 13-18 months(long-term).The short-term,medium-term and long-term curative effects were evaluated with unified curative criteria,and the results were compared with that obtained with the treatment of epidural injection of collagenase only.Results The short-term,medium-term and long-term effective rate of selected cases was 95.9%(519/541),90.4%(489/541)and 87.2%(472/541),respectively.When combined application of two methods was performed,different degree of shrinkage of the protruded nucleus pulposus was obviously observed,with a mean shrinkage degree of 30.5%.The short-term,medium-term and long-term effective rate of simple collagenase chemonucleolysis was 89.5%,82.4%and 80.4%,respectively.Conclusion Intradiscal injection of ozone combined with epidural injection of collagenase is an effective treatment for lumbar disc protrusion when the fibrous ring is broken,the protruded extent of nucleus pulposus is less than 10 mm and the dura sac and/or nerve root is compressed.Its curative effect is superior to simple collagenase chemonucleolysis.Ozone is of clinical significance for producing the shrinkage of the nucleus pulposus.