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.

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

BACKGROUND: Macrophage migration inhibitory factor (MIF) is mainly concerned with macrophage mobilizing function, as the upper stream cytokine of tumor necrosis factor-α(TNF-α), interleukin-1β (IL-1β), which may have critical effect in the process of the onset of rheumatoid arthritis. OBJECTIVE: To explore the correlations between the change of serum MIF and the activity of rheumatoid arthritis (RA) disease.DESIGN, TIME AND SETTING: Non-randomized control and case study, which was carried out in the Bethune International Peace Hospital of Chinese PLA from September in 2005 to October in 2006.PARTICIPANTS: Sixty RA patients were included in this study, and other thirty healthy subjects were selected as the control group. There were significant differences in age and sex between the two groups. METHODS: Clinical data of sixty RA patients were selected by carrying out retrospective analysis, then on the basis of disease activity score (DAS) accumulated points, they were divided into active and inactive group respectively, who were contrasted with 30 health adults. MAIN OUTCOME MEASURES: ① Morning stiffness (in minutes), joint tenderness index, arthrocele index, semi-quantity rheumatoid factor (RF), C-reactive protein concentration (CRP), erythrocyte sedimentation rate (ESR), and platelet count (PLT) were recorded; ② To compare the level of serum MIF, IL-1β and TNF-α among active group, inactive group, and control group; ③ The correlation analysis was carried out among the level of serum MIF, inflammatory index and clinical observation index.RESULTS: There was significantly increased in serum MIF of patients in the active group compared to of inactive and normal groups (P < 0.05), but there were no significantly differences between inactive and control groups (P > 0.05). There were significant correlations between the serum MIF concentration and active inflammatory index of RA disease, blood sedimentationrate (ESR), C-reactive protein (CRP), platelet counting (PLT), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), swell joint index (SJI) and tenderness joint, but no significant difference was observed between the serum MIF, age, disease course, morning stiffness and rheumatoid factor (RF).CONCLUSION: The serum MIF concentration is significantly increased in patients with RA, and it may be a useful parameter for monitoring disease activity of RA.

Objective:To investigate the feasibility of construction of tissue engineered cartilage by co-culture of bone marrow mesenchymal stem cells (BMSCs) and costal chondrocytes (CCs),and to provide theoretical basis and experimental basis for clinical repair of articular cartilage defects by Wuzhishan miniature pig knee cartilage defects with co-cultured cells.Methods:Density gradient centrifugation method was used to isolate BMSCs from Wuzhishan miniature pig.The double enzyme digestion method was used to isolate CCs.The passage 3 generation of BMSCs and passage 2 generation of CCs were randomly divided into 3 groups:a co-culture group of BMSCs∶CCs for 1∶2 (Group A),a simple CCs (Group B),and a simple BMSCs (Group C).The cell growth curve was drawn,and the content of glycosaminoglycan (GAG) of external separation in chondrocytes was determined.The 12 Wuzhishan miniature pigs were randomly divided into a co-culture cells/collagen membrane experimental group,a collagen membrane control group and the blank group.In the co-culture cells/collagen membrane experimental group,the co-cultured cells/collagen membrane were implanted into the cartilage defects of the mandibular condyle;in the collagen membrane control group,only collagen membrane was implanted;while in the blank group,nothing was implanted.Six animals were sacrificed at 8 and 16 weeks after surgery respectively (2 animals in each group).General observation,cartilage histological score and histopathological examination were carried out.Results:The BMSCs and co-culture cells grew well.The biological activity of CCs was good.After 16 weeks of operation,the repair tissues in the co-cultured cells/collagen membrane experimental group showed hyaline cartilage features:smooth,flat,and integrated well with the surrounding cartilage and subchondral bone.The collagen membrane in the collagen membrane control group was fibrously repaired.Repair tissue gross score in the co-culture cells/collagen membrane experimental group was significantly better than that in the collagen membrane control group and the blank group (both P<0.05),but there was no significant difference between the collagen membrane control group and the blank group (P>0.05).Conclusion:BMSCs,CCs and co-cultured cells can function as the seed cells for cartilage tissue engineering,and the co-culture cells (BMSCs∶CCs=1∶2) possess more advantages;the short-term effect of co-culture cells with collagen membrane on repairing cartilage defects is satisfied.

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.

Objective To discuss the changes of neutrophil gelatinase-associated lipocalin (NGAL) in patients with primary nephrotic syndrome (PNS)and the correlation with renal pathological type,renal tubulointerstitial lesions and the clinical indicators.Methods Forty patients with PNS were divided into acute kidney injury (AKI) group and non-AKI group according to whether renal tubular necrosis (ATN) occurred in renal pathology.Moreover,on the basis of pathological type they were divided into minimal change disease (MCD) group,mesangial proliferative glomerulonephritis (MsPGN) group,focal segmental glomerulosclerosis (FSGS) group,membrane proliferative glomerulonephritis (MPGN) group and membranous nephropathy (MN) group.Twenty healthy subjects and normal kidney tissues which came from 20 patients with renal tumor nephrectomy and were distant from the tumor sites were the control groups.Enzyme-linked immunosorbent assay (ELISA) was applied to detect the serum and urine level of NGAL,and immunohistochemical staining was used to observe the expression of NGAL in the renal tissue.Results (1)The serum and urine level of NGAL and the expression of NGAL in the renal tissue in the PNS complicated with AKI group were significantly higher than that in the PNS without AKI group and in the control group(P ＜ 0.05).(2)The serum and urine level of NGAL and the expression of NGAL in the renal tissue were enhanced in MPGN group and FSGS group than that in the other three groups(P ＜ 0.05).(3) Before developing to severe tubulointerstitial lesions,with the aggravation of tubulointerstitial damage,the serum and urine level of NGAL and the expression of NGAL in the renal tissue were increased.But when renal tubular interstitial lesions developed to severe disease,serum level of NGAL and the expression of NGAL in the renal tissue were decreased(P ＜ 0.05).(4)The serum and urine level of NGAL and the expression of NGAL in the renal tissue were positively correlated with serum creatinine(r values were 0.198,0.352,0.146 respectively,P values were 0.048,0.000,0.028 respectively),were positively correlated with blood urea nitrogen(r values were 0.199,0.278,0.325 respectively,P values were 0.043,0.000,0.019 respectively),were negatively correlated with serum albumin(r values were-0.384,-0.318,-0.259 respectively,P values were 0.028,0.024,0.020 respectively) and were negatively correlated with urine osmotic pressure(r values were-0.250,-0.256,-0.277 respectively,P values were 0.012,0.027,0.002 respectively).Conclusion NGAL is a sensitive biological parameter for predicting AKI in the patients with PNS,and it can be used to evaluate the degree of tubulointerstitial lesions and renal function to a certain extent.

Objective To investigate the role of silent mating type information regulation 2 homologue 1(SIRT1) in renal ischemia-reperfusion(IR) injury and its effect on NF-κBp65-peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) signal pathway in mice.Methods Seventy-two healthy C57BL/6 male mice were randomly divided into four groups:control group(n=18),sham-operated group(n=18),IR group(n=18),resveratrol group(n=18).Bilateral renal pedicle were clamped for 45 min was adopted to establish the model of acute ischemic renal injury,to give 2％ dimethyl sulfoxide or resveratrol by intraperitoneal injection for 7 days before modeling.Determination techniques included routine biochemical methods for the the levels of Scr and BUN,spectrophotometry for the level of superoxide dismutase (SOD),HE staining for the histological changes as well as immunohistochemical method and Western blotting for the expressions of SIRT1,NF-κBp65 and PGC-1α,respectively.Results Compared with that in control and sham-operated groups,the levels of serum Scr and BUN were higher and SOD levels in renal tissues were lower at 12 h and 24 h after operation in IR groups(P ＜ 0.05).HE staining revealed evident pathological lesions including necrosis of renal tubular epithelial cells in IR group.Compared with that in IR group,resveratrol attenuated the above-mentioned changes.Western blotting revealed the up-regulated SIRT1 expression and the activated NF-κB signal pathway,the up-regulated p65 expression and the down-regulated PGC-1αexpression subsequent to IR(P ＜ 0.05).Both Western blotting and immunohistochemistry showed that the expressions of SIRT1 and PGC-1α in resveratrol group were up-regulated compared to that in IRgroup(P ＜ 0.05),while the NF-κBp65 expression in resveratrol group was down-regulated(P ＜ 0.05).Conclusions In mouse model of renal ischemia-reperfusion injury,the activation of SIRT1 can inhibit the NF-κBp65 expression and accordingly up-regulated PGC-1α level,contributing to inhibiting inflammatory reactions and attenuating oxidative stress-induced injury in the protection of the kidneys.

Objective To study the significance of expressions of smad_4mRNA,TGF-?_1, and TGF-?R_1 in pancreatic carcinoma(PC) . Methods Smad_4mRNA was detected by in situ hybridization. TGF-?_1 and TGF-?R_1 were detected by immunohistochemical method. Results The positive rates of smad_4mRNA,TGF-?_1 and TGF-?R_1 were singnificantly lower in 53 slices of pancreatic carcinoma than those in 25 slices of paracancerous tissue (all P

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.

BACKGROUND:Articular chondrocytes with the ability of autocrine and paracrine can provide the growth factors and microenvironment for synovial mesenchymal stem cels differentiating into the chondrocyte. The
three-dimensional scaffold could provide space for cels adhesion, proliferation and differentiation.
OBJECTIVE: To study the ability of chondrogenesis by co-culturing synovial mesenchymal stem cels and chondrocytes under the three-dimensional condition.
METHODS:The synovial membrane and articular cartilage were harvested from rat knee joint. The synovial
mesenchymal stem cels and chondrocytes were obtained through the method of enzyme digestion. The passage 3 synovial mesenchymal stem cels and passage 2 chondrocytes were co-cultured in the chitosan/I colagen
composite scaffolds at the ratio of 1:2. Then, the cels/scaffold composite was harvested to be examined
morphologicaly, histologicaly and immunohistochemicaly after being cultured 21 days. The confocal laser was also employed to detect the cels distribution in the scaffold.
RESULTS AND CONCLUSION: After being cultured 72 hours, it could be observed from the cels/scaffold composite examined through the scanning electron microscope that the cels adhered on the surface of the
scaffold and extracelular matrix surrounding the cels was seen on the scaffold. After being cultured 21 days, it could be found through the confocal laser scanning that the cels were wel-distributed on the scaffold, and cels decreased gradualy. Type II colagen was positive in the extracelular matrix immunohistochamicaly. It
suggested from this study that the synovial mesenchymal stem cels could be co-cultured with chondrocytes in the chitosan/I colagen composite scaffolds and have the ability of chondrogenesis differentiation.