ABSTRACT
In the present paper, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) was used to examine and identify differentially expressed genes in Capsicum annuum exposed to UV-B irradiation. Around 4000 transcript derived fragments (TDFs) were visualized and in total 183 TDFs were isolated, sequenced and analyzed by Blast 2 go. Among these TDFs, 84 of them showed homology to known genes. There were 43 TDFs showing up-regulated expression, 24 TDFs showing down-regulated expression and 27 TDFs showing both up-regulated and down-regulated expression, respectively. Some of these TDFs were found to be in response/related to UV-B stress, including carbonic anhydrase, calcium-dependent protein, thionin-like protein, bzip protein and so on. In particular, chlorophyll a/b binding protein (Capcab) responding to UV-B stress was cloned. It was concluded that Capcab could play a protective role in plant anti-UV-B and maintaining photosynthetic rate under UV-B stress.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of different cell lysis buffers on protein quantification with Bradford method and bicinchoninic acid (BCA) method.</p><p><b>METHODS</b>Bradford method and BCA method were used to determine the concentration of bovine serum albumin (BSA) in different solutions (distilled water, cell lysis buffer used in two-dimensional differential in-gel electrophoresis and three kinds of cell lysis buffers used in conventional two dimensional gel electrophoresis), as well as the protein concentrations of cell lysates using these different lysis buffers. Bradford method was also applied to determine the protein concentrations of samples with repeated freeze thaw cycle, in different colorimetric cylinders, or using different standard curves from different periods.</p><p><b>RESULT</b>The protein measurements increased for 1.2 to 2 fold when different cell lysis buffers were used in Bradford method, but the measurements increased with the increased concentration of BSA (r=0.989 approximately 0.996, P<0.05). For BCA, measurement reading increased about thousands times higher, even overflowed the limits of machine. Protein measurements didn't change significantly, only showed a declined trend after repeated freeze thaw cycle, while no significant changes were found using different colorimetric cylinders or standard curves from different periods.</p><p><b>CONCLUSION</b>Bradford method may be the choice of the protein quantification in proteomics. However, optimization is required for specific experimental conditions.</p>
Subject(s)
Buffers , Cells , Chemistry Techniques, Analytical , Methods , Proteins , Serum Albumin, Bovine , Spectrophotometry, UltravioletABSTRACT
<p><b>OBJECTIVE</b>To analyze the complications and treatment results of intraoperative radiotherapy (IORT) for esophageal carcinoma.</p><p><b>METHODS</b>Sixty patients with thoracic esophageal carcinoma underwent esophagectomy through right thoractomy, 30 patients of whom received IORT of 15 - 25 Gy.</p><p><b>RESULTS</b>In patients who underwent IORT, 2 cases of pneumonitis, 1 case of anastomotic leak and 1 case of incisional wound infection were found. In patients underwent surgery only, 1 case of thoracic empyema and 1 case of anastomotic leak were found. All the complications ultimately healed. There was no operative mortality. During the follow-up of 3 years, in patients who underwent IORT, 2 of 3 died of radiation pneumonitis 24 and 26 months after IORT with one complicated with bronchoesophageal fistula. One of 3 died of multiple lung metastases. The 3-year survival rate was 88.0% (22/25) in IORT group and 76.0% (19/25) in surgery only group.</p><p><b>CONCLUSION</b>Intraoperative radiotherapy can reduce locoregional recurrence if performed to thoracic esophageal carcinoma patients without surgical contraindication or distant metastasis. Radiation pneumonitis, a common complication difficult to manage, implies a poor prognosis and, consequently, the lung and bronchus should be protected from the radiation.</p>