ABSTRACT
Objective To construct the ?-gal reporter genes containing the 5′-end flanking of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) gene in different sequence lengths and identify the sequence, which regulates the gene expression of EOLA1 by the ?-gal analysis system. Methods The target sequences were amplified by the method of genome walker, and were inserted into the upstream of ?-gal gene located in the ?-gal enhancer vector by the directional clone technique respectively; the regulative sequence was identified by analyzing the ?-gal activities of reconstructed plasmid in ECV304 cells. Results The regions, containing 2 659 bp and 1 951 bp upstreaming from exon 1, significantly stimulated the reporter gene activity as compared with that of the ?-gal control vector in transfected cells. But the region, containing 361 bp upstreaming from exon 1, did not stimulate the reporter gene activity. Conclusion There is an up-regulative element of gene transcription in the region of -361 to -1 951 bp in EOLA1 gene upstream.
ABSTRACT
In order to explore the effects of exogenous human telomerase reverse transcriptase (hTERT/hTRT/hEST2) on telomeric restriction fragment (TRF), telomerase activity and its subunits expression in human embryonic fibroblasts (hEFs), hTERT sense eukaryotic expression vector pIRES2 EGFP hTERT was constructed with DNA recombinant technique and then transfected into primary hEFs by Lipofectin method. TRF length, telomerase activity and changes in telomerase subunits expression were examined and evaluated in transfected and untransfected cells. The results showed that telomerase activity in pIRES2 EGFP hTERT transfected cells (hEF EGFP) was significantly higher than that in untransfected hEFs and vacant vector transfected cells (hEF EGFP) ( P
ABSTRACT
OBJECTIVE: This study aimed at understanding of the role of calcium homeostasis in cardiomyocytes from hypoxia, burnt serum-induced injury. METHODS: Alterations in cytosolic free calcium concentration (Ca(i)), calcium influx and viability of the cardiomyocytes in vitro after hypoxia, burnt serum stimulus were observed. RESULTS: Ca(i) increased markedly, in the meantime, the cellular transmembrane calcium influx increased and the viability of the cells decreased significantly following hypoxia, burnt serum-induced injury. CONCLUSIONS: In our study cytosolic calcium ion was transported abnormally in the cardiomyocytes after burn, to result in Ca(i) increase and runaway calcium homeostasis, thus the normal cellular function was disturbed. This may be one of the important factors in the development of burn-induced cardiac injury.
ABSTRACT
The results of early resuscitation of 104 adult patients with BSA more than 50%were studied retrospectively. Although the mean amount of fluid replaced during 48h postburn was similar to the amount calculated with our formula, there existed significant individual differences. Therefore it seems not necessary to set up a rigid fluid replacement plan. To ensure adequate tissue perfusion, the fluid replacement formula might be modified as follows: 2 ml/kg/BSA%, with urinary output 30-40 ml/h, in the first 24h; and 1.5ml/kg/BSA%, with urinary output 40-50ml/h, in the second 24h. It should be emphasized that resuscitation should be started as early as possible,and adequate amount of fluid replacement is especially important during the first 2-3h postburn.There was no obvious relationship between the incidence of visceral complications and the total amount of fluid replaced during the resuscitation. Available data indicated that the amount of fluid calculated on the basis of our formula neither increased the incidence of early pulmonary edema nor influenced its development.As far as prevention of pulmonary edema was concerned, it did not seem justifiable to restrict the amount of resuscitation fluids. It was also noted that fluid therapy alone would not prevent entirely the development of postburn renal insufficiency.