Objective To investigate the effects of norepinephrine on brain edema of rats in 24 h after severe burn.Methods A total of 48 healthy Wistar rats were randomly divided into 8 groups: normal control group,1,2,5 mg/kg norepinephrine,burn group,burn with 1,2,5 mg/kg norepinephrine pretreatment groups(n=6 in each group).The rats in all burn groups were scalded into 40%TBSAⅢ degree burn.Pathological features were observed,and blood brain barrier,brain water(%) were examined in postburn 24 h.Results Pathological evidence of brain edema exhibited in the burn group and burn group with norepinephrine pretreatment,and increased permeability of blood brain barrier and brain water were observed.The burn with norepinephrine pretreatment groups were more significantly severe in comparison with simple burn groups and normal control group.Conclusion Norepinephrine may play an important role in brain edema in postburn 24 h,suggesting that stress of postburn may induce brain edema.

Objective To investigate the effects of norepinephrine (NE) on vascular endothelial growth factor (VEGF) expression of brain tissues in severe burn rats. Methods The healthy male Wistar rats were made into 40%TBSAⅢ?burn models to observe the effect of NE on blood brain barrier. In the meantime, effect of NE was examined by means of immunocytochemistry and real time PCR. Results (1) Permeability of blood brain barrier was increased in burn and burn with NE stimulating rats, with significantly statistical difference compared with normal control group (P

To explore the role of apoptosis, apoptosis regulating genes in the pathogenesis and development of smoke inhalation injury. With smoke inhalation injury rat model, the changes int the expression of Bcl 2, Bax, Fas, FasL genes mRNA and protein contents and their relationship with apoptosis of lung tissue cells at different time points after the injury were observed with TUNEL, immunohistochemistry and RT PCR techniques. The results showed that: ①apoptosis indet of lung colls after smoke inhalation injury increased, ②expressions of Bcl 2, Bax, Fas, FasL genes were obviously up regulated in injury group, peaking at the 12th hour, whereas the peak of protein expression was at the 24th hour. Furthermore, a significant correlation was found between the expression of Fas, Fasl, Bax gene and apoptosis indices in lung cells. The results suggested that apoptosis participated in the early pathological process of smoke inhalation injury, and apoptosis regulating genes foot part in the regulation of apoptosis in smoke inhalation injury.

AIM: To investigate the changes of gene expression of vascular endothelial growth factor(VEGF) in norepinephrine/ burn serum-induced astrocytes.METHODS: Immunofluorescence staining was used to show the distribution of VEGF in astrocytes after 24 h using norepinephrine/burn serum stimulation.Western blotting was used to detect protein expression of VEGF.Real time PCR was used to investigate expression of VEGF mRNA.RESULTS: ① Green fluorescence of protein expression of VEGF in astrocytes was increased when treated with high dose norepinephrine(50 ?mol/L).Green fluorescence of protein expression of VEGF in astrocytes was increased distinctness after burn serum stimulation.Green fluorescence protein expression of VEGF in astrocytes was increased significantly when high dose norepinephrine combined with burn serum stimulation was added.② VEGF protein expression in burn serum stimulating group was increased,and VEGF protein expression was significantly increased when burn serum was added during moderate(20 ?mol/L),high dose norepinephrine stimulation.③ Expression of VEGF mRNA was increased in burn serumtreated astrocytes.Expression of VEGF mRNA was increased significantly when norepinephrine-stimulated astrocytes exposed to barn serum,and as norepinephrine dose increases gradually.CONCLUSION: Norepinephrine and burn serum play an important role in inducing VEGF protein expression in astrocytes,suggesting that stress reaction of postburn is an important cause in inducing brain edema by excreting VEGF in astrocytes.

AIM: To investigate the effect of norepinephrine (NE) on brain tissue VEGF protein expression in 24 h after severe burns. METHODS: (1) 40% total body surface area (TBSA) Ⅲ? burn model was made in Wistar males rats. Brain water (%) was examined at 24 h after burn. (2) NE levels in the rat brain tissue were determined by high performence liquid chromatography. (3) VEGF protein levels in the rat brain tissue were determined by Western blotting at 24 h of postburn. RESULTS: (1) Brain edema exhibited at the burns and burn with norepinephrine groups. (2) A increase in NE levels in the rat brain tissue was observed at the burns. NE levels increased significantly in burn with norepinephrine groups. (3) VEGF protein expression in the rat brain tissue was gradually increased with increasing in norepinephrine stimulating dose. VEGF protein expression in the burn rats brain tissue was significantly increased in burn with high dose of norepinephrine stimulation. CONCLUSION: Norepinephrine induced VEGF protein expression in rat brain tissue at 24 h after severe burn, suggesting that increases in norepinephrine level postburn induce brain edema.

Objective To investigate effects of the complex therapy on severe inhalation injury. Methods The cases with inhalation injury and acute respiratory failure (ARF) of 4172 consecutive burn patients from Jan. 2000 to Feb. 2003 in our institute were studied retrospectively. Results 30 of the 128 patients with inhalation injury occurred acute ARF. 48 hours after the exogenous pulmonary surfactant (PS) was used to treat 6 cases with uncomplicated ARF, pulmonary function obviously ameliorated as compared with routine therapy. PaO 2 /FiO 2 rose from 239?33 mmHg to 317?28 mmHg(P

Objective To explore a suitable plan for the delayed rapid fluid resuscitation in burn patients with shock. Methods 20 patients with total body surface area (TBSA) burned over 40% admitted 4～8h after postburn were enrolled in this study. The patients were randomly divided into plasma and gelofusin groups. Rapid fluid replacement was given immediately after admission under close hemodynamic monitoring. Hemodynamic (PAP, PAWP, CO, PVR, SVR) and hemorrheological parameters, tissue oxygenation (DO 2, VO 2, O 2ext , lactic acid, base deficit) as well as indices reflecting the main visceral functions and damage were investigated. Results The amount of rapid fluid infusion within 2h after admission accounted for 38 8?6 1% of the amount calculated with the formula for the first 24h. When the infusion amount of pre-hospitalization was added, the amount would be (48 3?5 0)% of the amount for the first 24h. The real amount of the infusion for the first 24h was (31 4?14 3)% more than that of the amount calculated with the Evans formula. The real infused fluid amount for the second 24h was almost equal to the amount calculated with the formula. After fast fluid replacement therapy, all the parameters determined were markedly improved. Conclusions It is proposed that the formula for the delayed rapid fluid resuscitation in burn patients with shock should be: the amount infused for the first 24h (ml) =TBSA (%)?body weight (kg)?2 6,the ratio of colloid to electrolytes is 1:1, water=2000ml. Half of the total amount should be infused in the first 2h after admission under close hemodynamic monitoring. The amount infused for the second 24h (ml)=TBSA (%)?body weight (kg)?1,the ratio of colloid to electrolytes is 1:1, water=2000m1.

Objective To study the subcellular localization and the tissue expression of EOLA1(endothelial-overexpressed lipopolysaccharide-associated factor 1). Methods The fusion protein EOLA1-EGFP expressed vector was constructed and transfected into endothelial cells. After 24 hours posttransfection, the subcellular localization of EOLA1 was detected by laser-scanning microscopy. The tissue-specific distribution of EOLA1 was assessed with Multiple Tissue Northern Blots. Results The expression of EOLA1 was tissue-specific in various human tissues. With human multiple tissue Northern blot analysis, it was shown that EOLA1 could express in the heart, skeletal muscle, kidney, liver, placenta, colon, spleen, small intestine, but did not in the brain, lung, thymus, and peripheral blood leukocyte. EOLA1 was mainly localized in cytoplasm and could move into the nucleus. Conclusion EOLA1 is one of intercellular proteins and may play a role in intercellular signal transduction.

Objective To construct small interfering RNA(siRNA) expression vectors targeting EOLA1 (Endothelial-overexpressed lipopolysaccharide-associated factor 1) gene, and to assay their effects on the expression of EOLA1. Methods GFP-EOLA1 fusion protein expressive vector pEGFP-N2/EOLA1 was constructed and transfected into ECV304 cells. The transfected cells were cultured in M199 containing G418 (400?g/ml) for 5 weeks to screen the cell line stably expressing GFP-EOLA1 fusion protein. For interfering EOLA1 target gene, complementary oligonucleotides were desiqned and synthesized at different sites. The oligonucleotides were inserted into pSinencer3.1/H1 plasmid. Then, the vector was transfected into the ECV304 cells stably expressing GFP-EOLA1 fusion protein and the inhibiting affection to target gene EOLA1 was identitied by the observation of the green fluorescence in transfected cells with inverted fluorescent microscope and Western immunoblot assay. Results The ECV304 cell line stably expressing GFP-EOLA1 fusion protein was constructed, and the siRNA vector of EOLA1 can knockdown EOLA1 gene expression specifically. Conclusion The siRNA vectors specifically interfering EOLA1 expression were constructed successfully, which would be useful in the study of function of EOLA1 gene.

Objective To screen and analyze genes up regulated in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS). Methods Suppression subtractive hybridization (SSH) was performed between the unstimulated HUVEC(driver) and HUVEC stimulated with LPS(tester) to generate subtractive cDNA library. The library was screened with colony dot hybridization to further verify the differentially expressed cDNA clones. Positive clones were sequenced and BLAST analyzed. The 3 novel cDNA sequences were verified by RT PCR. Results Twenty five up regulated genes related to inflammation, cellular cytoskeletal rearrangement, cellular proliferation and apoptosis, intercellular message transduction, and 3 new expression sequence tags (EST) were acquired. RT PCR indicated the expression of the new ESTs only in HUVEC stimulated by LPS. Conclusion SSH is a powerful technique of high sensitivity for the detection and clone of up regulated gene expressed in HUVEC stimulated by LPS, which may be helpful to clarify the mechanism of endothelial cells activation stimulated by LPS.