Chinese Journal of Biotechnology,Vol 34,No. 02
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 p ET-21 a, 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.
Quantitative proteomics analysis of isoniazid, streptomycin mono-drug resistant and poly-drug resistant clinical isolates of Mycobacterium tuberculosis
Acta Microbiologica Sinica,Vol 58,No. 07
[Objective] To investigate the differentially expressed proteins between isoniazid, streptomycin mono- and poly-drug resistant clinical isolate strains of Mycobacterium tuberculosis. [Methods] Cellular proteins were extracted from isoniazid, streptomycin mono- and poly-drug resistant clinical isolates M. tuberculosis and H37Rv. Differentially expressed proteins were identified through isobaric tags for relative and absolute quantification (iTRAQ) combined with Nano LC-MS/MS technology. The biological function and interaction among differentially expressed proteins and isoniazid or streptomycin were analyzed by DAVID 6.7 and STITCH 5.0, respectively. [Results] 58 differentially expressed proteins were identified in isoniazid, streptomycin mono-drug resistant strains and isoniazid, streptomycin poly-drug resistance strain compared with the proteomic profiles of H37Rv. The biological function of differentially expressed proteins mainly relates to oxidoreductase and transferase activity. Compared with isoniazid and streptomycin poly-drug resistance strain, two proteins (Rv2986c and Rv1908c) were up-regulated over 1.25 folds and two proteins (Rv3133c and Rv0577) were down-regulated less than 0.7 fold in isoniazid, streptomycin mono-drug resistant strains. Bioinformatics predicted that the four proteins interact with isoniazid and streptomycin directly or indirectly. [Conclusion] The expressed level or the interaction together of Rv2986c, Rv1908c, Rv3133c and Rv0577 is likely to be related to isoniazid and streptomycin resistance in M. tuberculosis.
Acta Optica Sinica,Vol 37,No. 04
Wedge and strip anode (WSA) is one of the charge division anodes. In the photon counting imaging detector, the role of the WSA is to decode the incident position of photon events. The anode performance parameters have an important influence on the imaging performance of the detector. The inter-electrode capacitance of WSA is studied, and its calculation formula is obtained by the theoretical model. The WSA panel with different parameters is designed and prepared. The relationship of cycle length, insulation gap width, anode collecting area, and substrate material with the anode inter-electrode capacitance is analyzed. Test results show that the inter-electrode capacitance is in accordance with the theoretical calculation result and the deviation between calculated value and test value is within 10%.
Clinical observation of wrist–ankle needle therapy combined with patient controlled intravenous analgesia for pain after laparoscopic surgery for eccyesis
Chinese Acupuncture & Moxibustion,Vol 37,No. 11
Objective To observe the effect difference between wrist–ankle needle therapy combined with patient controlled intravenous analgesia (PCIA) and simple PCIA for pain after laparoscopic surgery for eccyesis. Methods Ninety-eight patients were assigned into an observation group and a control group by random number table, with 49 cases in each one. Intravenous-inhalational anesthesia was used in the two groups. Simple PCIA for pain was applied in the control group. wrist–ankle needle therapy at bilateral ankle areas 1 and 2 combined with PCIA were implemented in the observation group. The pain state of incision was recorded based on visual analogue scale (VAS) at 1 h, 2 h, 6 h, 12 h, 24 h, 36 h and 48 h after surgery. The total effective rates and adverse reaction rates within 48 h after surgery were compared between the two groups. Results The VAS scores at 6 h, 12 h and 24 h after surgery in the observation group were lower than those in the control group (all P < 0.01), and the scores in the other time points were not statistically different (all P > 0.05). The total effective rate of the observation group was 98.0% (48/49), which was better than 83.7% (41/49) of the control group ( P < 0.05). The adverse reaction rate of the observation group was 12.2% (6/49), and that of the control group was 69.4% (34/49), with statistical difference ( P < 0.01). Conclusion wrist–ankle needle therapy combined with PCIA can effectively relieve pain after laparoscopic surgery for eccyesis, and reduce adverse reaction rate after surgery.
Acta Optica Sinica,Vol 37,No. 05
In view of the inconsistent perspective in target images obtained by pinhole array for traditional X-ray gated micro channel plate (MCP) framing camera, an implement method of X-ray framing camera with single perspective is proposed. At the electro-optic intersection in electrostatically focused image tube, the electrons carry holographic information, and can be divided into multiple output images with single perspective; subsequently, a gated MCP unit is matched with the frame to enhance the target image. Compared with the traditional gated MCP framing camera, the designed framing camera realizes identity of image perspective and avoids direct light’s excitation on screen, and a higher signal-to-noise ratio is obtained. The designed framing camera has an effective input working diameter of 40 mm, an output image diameter of 40 mm, a magnification of 1.29, the center spatial resolution of 60l p/mm in theory, the marginal spatial resolution of 26 lp/mm, and the geometric distortion of less than 15%. Single-frame exposure duration is determined by shutter pulse time of gated MCP framing unit. This method is an effective way to realize identity of image perspective for X-ray framing camera.