5 days.Blood gas analysis and blood pressure were determined at admitted day.Meanwhile,peripheral white blood cells at d1,3,5,and blood glucose were measured every day,respectively.GCS at d1,3,5 and hyperglycemia scorce(HS) were evaluated.Results Of the 82 studied patients,36 cases died.Univariate analysis showed that hypotension,lower GCS,higher peripheral white blood cells and HS were the independent death risk factors(Pa0.05).In multivariate logistic regression,the factors significantly associated with an increase in mortality were hypotension,lower GCS and higher HS.Conclusion Lower GCS,higher HS and hypotension are associated with poor outcome of children with severe trauma brain injury.

Acupuncture Therapy ; Altitude ; Animals ; Brain Edema ; prevention & control ; Female ; Hypoxia, Brain ; therapy ; Male ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Adenosine A1 ; metabolism

Adult ; Diagnosis, Differential ; Female ; Humans ; Pancreas ; abnormalities ; Steatorrhea ; etiology

OBJECTIVETo analyze the transcription activation and possible regulation mechanism of human X-box binding protein 1(XBP1)gene 5'upstream DNA sequence in different cell lines.

METHODSSix kinds of XBP1 promoter deletion mutants were cloned into pGEM-Teasy vector, which included XBP1 gene 5' upstream -1039 to 66 bp,-859 to 66 bp,-623 to 66 bp,-351 to 66 bp,-227 to 66 bp,-227 to -45 bp respectively. Every deletion mutant sequence was cut from Teasy-XBP1p by KpnI and Xho I, and subcloned into pCAT3-Basic to produce a set of constructs termed as p1-XBP1p, p2-XBP1p, p3-XBP1p, p4-XBP1p, p5-XBP1p, p6-XBP1p, respectively. The transcription activity of each construct was detected after transiently transfecting K562, HepG2,NIH-3T3 and L0(2)cell with FuGENE 6 transfection reagent. Cells transfected by pCAT3-Basic or pCAT3-Promoter were used as negative and positive controls. The activity of chloramphenicol acetyltransferase(CAT), which reflects the transcription activation of the XBP1 gene promoter, was detected by ELISA after 48 hours of transfection.

RESULTSThe reporter vectors of six kinds of XBP1 promoter deletion mutants were successfully constructed, as confirmed by restriction enzyme digestion and sequencing. The activities of p4-XBP1p and p5-XBP1p were higher than the other deletion mutants in K562 and HepG2. And the activity of p5-XBP1p was the highest in HepG2. There was no activity detected from any transfected NIH-3T3.

CONCLUSIONThe XBP1 gene promoter can transactivate its downstream gene to transcription. The core sequence of XBP1 promoter was implied between -227 bp and 66 bp. This sequence was connected with the transcriptional activity of XBP1 promoter closely. Its transcription activity varies with different cell lines. XBP1 promoter might drive gene expression with cell-type specificity.

3T3 Cells ; 5' Flanking Region ; genetics ; Animals ; Base Sequence ; Cell Line ; Chloramphenicol O-Acetyltransferase ; metabolism ; DNA ; analysis ; DNA-Binding Proteins ; genetics ; Gene Deletion ; Gene Expression Regulation ; physiology ; Genes, Reporter ; Humans ; K562 Cells ; Mice ; Molecular Sequence Data ; Nuclear Proteins ; genetics ; Promoter Regions, Genetic ; genetics ; Regulatory Factor X Transcription Factors ; Transcription Factors ; Transcription, Genetic ; physiology ; Transcriptional Activation ; Transfection ; Tumor Cells, Cultured ; X-Box Binding Protein 1

OBJECTIVESince continuous blood purification (CBP) has the effects of eliminating inflammatory mediators and improving organs function, CBP had been applied to treat non-renal diseases for nearly 10 years, but few studies have been conducted in children with sepsis and multiorgan dysfunction syndrome (MODS), especially in China. The present study aimed to evaluate the clinical effect of CBP in treatment of children with severe sepsis and MODS.

METHODSTwenty-two children with severe sepsis and MODS admitted to our PICU from Aug. 2003 to Aug. 2005 were treated with continuous veno-venous hemodialysis filtration. Their heart rate, arterial blood pressure, doses of vasoactive agents, spontaneous respiratory rate, PO2/FiO2 and prognosis were investigated.

RESULTSCatheterization and CBP were carried out in all the 22 children. Continuous vein-vein hemodialysis filtration (CVVHDF) and pre-dilution were chosen. The duration of CBP was (64.4 +/- 34.5) h. All the children had tachycardia before CBP and the heart rate fell gradually to 45 +/- 13 bpm 4 h after CBP. Blood pressure (BP) was stable in 7 children without shock during CBP. Ten children with early shock could maintain normal BP during CBP, but the doses of vasoactive agents were tapered 1 to 5 h after beginning of CBP and use of these agents was discontinued at 2 to 8 h. BP was elevated by (25.2 +/- 10.7) mmHg (1 mmHg = 0.133 kPa) in 5 refractorily shocked children 4 h after CBP and returned to normal level 8 h later. The doses of the vasoactive drugs were reduced at 2 to 8 h and ended 4 to 16 h later, which was longer than that of children with early stage shock. The accelerated spontaneous respiratory rate was slowed down by 7 +/- 4 per minute 4 h later, PO2/FiO2 rose from (177.7 +/- 53.1) mmHg before CBP to (341.0 +/- 60.2) mmHg 4 h after CBP in children with respiratory failure and reached the normal value (5.3 +/- 2.1) h later. FiO2 declined to less than 50%. Pediatric critical illness score was 62.2 +/- 7.4 on admission and elevated to (86.6 +/- 9.0) 24 h later, which was a significant elevation as compared to that of children with sepsis who were not treated with CBP seen between Aug. 2001 and July 2003. The survival rate was 72.7% after CBP and the effective rate of the treatment was 90.9%, but was 36% in children who were not treated with CVVHDF.

CONCLUSIONCBP can effectively improve the vital organ's function of children with sepsis and MODS and raise their survival rate. Replacement fluid of modified Ports formula was useful for stability of serum potassium and sodium, but resulted in elevation of serum glucose, calcium, and osmolarity. The application of CBP in children with sepsis can lead to slight drop of blood pressure at the beginning and to bleeding during CBP.

Adolescent ; Blood Pressure ; Child ; Child, Preschool ; Female ; Heart Rate ; Hemofiltration ; methods ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Pediatric ; Male ; Multiple Organ Failure ; etiology ; mortality ; physiopathology ; therapy ; Prognosis ; Sepsis ; complications ; mortality ; physiopathology ; therapy ; Severity of Illness Index ; Survival Rate ; Treatment Outcome

OBJECTIVEMost of the therapeutic strategies for systemic inflammatory response syndrome (SIRS) is not effective. This study was to investigate the effect of continuous veno-venous hemodiafiltration (CVVHDF) on SIRS induced by cecum perforate peritonitis in piglets.

METHODSTwelve piglets (weighing 7-9 kg) were randomly divided into two groups: control and CVVHDF (n=6). The piglets of both groups were subjected to a cecum puncture to induce peritonitis which caused SIRS. After SIRS occurred the piglets of the CVVHDF group immediately received the CVVHDF therapy for 6 hrs, with a blood flow rate of 20 mL/min, a replacement rate of 300 mL/h, and a dialysis rate of 600 mL/h. The heart rate (HR), mean artery blood pressure (MABP), respiratory rate (RR), arterial blood gas analysis and blood cells count were measured and recorded at baseline and onset of SIRS, and 2, 4 and 6 hrs after SIRS occurred.

RESULTSWhen SIRS occurred, the HR and RR increased and the MABP, artery oxygen pressure (PaO2) and the count of white cells decreased in both groups. The HR of the CVVHDF group decreased significantly at 2 hrs (P < 0.05) and remained lower until 6 hrs after CVVHDF therapy (P < 0.01) compared with that of the control group. The RR of the CVVHDF group was significantly lower than that of the control group 6 hrs after CVVHDF therapy (P < 0.05). The MABP of the CVVHDF group increased significantly 4 and 6 hrs after therapy compared with that of the control group (P < 0.01, P < 0.05 respectively). There were no significant differences in temperature, PaO2 and blood cells count between the two groups during the experiment.

CONCLUSIONSCVVHDF has a positive effect on hemodynamics in piglets with SIRS induced by cecum perforate peritonitis.

Animals ; Blood Pressure ; Body Temperature ; Carbon Dioxide ; blood ; Cecum ; injuries ; Central Venous Pressure ; Female ; Heart Rate ; Hemodiafiltration ; Intestinal Perforation ; complications ; Male ; Oxygen ; blood ; Peritonitis ; complications ; Swine ; Systemic Inflammatory Response Syndrome ; therapy

OBJECTIVETo investigate the effect of continuous veno-venous hemodiafiltration (CVVHDF) on endotoxin-induced acute lung injury (ALI) of piglets.

METHODSEighteen piglets were randomly divided into three groups: control group (n = 6); heparin group (n = 6) and CVVHDF treatment group (n = 6). All the animals were anesthetized by muscle injection of ketamine (30 mg/kg), then placed in supine position, received continuous intravenous infusion of ketamine with the rate of 10 mg/(kgxh). After placing a 4.5 cm (inner diameter) tracheal tube via tracheostoma, controlled mechanical ventilation was established using the assisted-controlled ventilation option of the NEWPORT 200. Respiratory rate at 30 breath/min; PIP at 10 cm H2O (1 cm H2O = 0.098 kPa); PEEP at 2 cm H2O and fraction of inspired oxygen at 0.3. A vein catheter was placed into right vena jugularis interna to administer a Ringer's solution. Initially, at a rate of 10 ml/kg, followed by a rate of 15 ml/kg when the mean arterial blood pressure was below 70 mm Hg (1 mm Hg = 0.133 kPa), the rate of 20 ml/kg was used when the mean arterial blood pressure was below 60 mm Hg. An 8Fr double-lumen catheter was inserted into left femoral vein and served as the pathway for CVVHDF. A Pulsiocath Pcco catheter was positioned into left femoral artery to monitor the circulatory parameters. All catheters were flushed with heparinized saline to prevent clotting. Then all the animals were given intravenous infusion of 150 microg/kg endotoxin within 30 minutes to induce ALI. When the oxygenation index < 300 and pulmonary compliance < 30% of the baseline, the animals of heparin group received heparin infusion to maintain blood active coagulation time (ACT) 180 - 250 s, the animals of treatment group received CVVHDF with the blood flow of 50 ml/min, replacement rate of 300 ml/h, dialysis rate of 600 ml/h and the ultrafiltrate rate of 350 ml/h for six hours, heparin infusion to keep blood ACT 180 - 250 s. The circulatory parameters: heart rate (HR), mean arterial blood pressure (MABP), central venous pressure (CVP), pulse contour cardiac output index (PCCI); systemic venous resistance index (SVRI), cardiac function index (CFI), external venous lung water index (EVLWI), left ventricular contractile index (dPmx); respiratory parameters: respiratory rate (RR), pulmonary compliance (Cdyn) were monitored; arterial blood gas analysis was performed and oxygenation index (PaO2/FiO2) was calculated. All the parameters were recorded at baseline (B), onset of ALI (A 0 h), two hours (A 2 h), four hours (A 4 h), six hours (A 6 h) after ALI.

RESULTSNo significant difference in circulatory parameters, respiratory parameters and blood gas analysis were found at B and A 0 h among the three groups. When the ALI occurred, PaO2/FiO2, Cdyn, MABP and PCCI of the three groups decreased; HR, RR, EVLWI, SVRI increased. After four hours of ALI, the RR, EVLWI, SVRI, CFI and dPmx of treatment group were improved, the differences were significant compared with the other two groups (P < 0.05). After six hours of ALI, the HR, PCCI, MABP, PaO2/FiO2 and Cdyn of treatment group were significantly improved, compared with control group and heparin group (P < 0.05). There were no significant differences in any of the parameters between control group and heparin group. The difference in CVP among three groups was not significant.

CONCLUSIONCVVHDF has a good effect on hemodynamics of the endotoxin-induced ALI of the piglets.

Acute Lung Injury ; etiology ; physiopathology ; therapy ; Animals ; Endotoxins ; adverse effects ; Hemodiafiltration ; Hemodynamics ; Swine

To study effect of pre-freezing temperature and lyophilizer shelf temperature on recovery of human red blood cells after lyophilization and determine solidifying temperature of this lyophilization system, the protective solution composed of 7% DMSO, 40% polyvinylpyrrolidone (PVP) and isotonic buffer were adopted to lyophilize red blood cells at different pre-freezing temperatures or shelf temperatures. At first, fresh whole blood was centrifugated, washed and equilibrized to prepare concentrated red blood cells. Then concentrated red blood cells were mixed with the protective solution at 1:3 and pre-freezed at different temperature (-20, -35, -45, -80 or -196 degrees C) before lyophilization in lyophilizer. To study effect of shelf temperature on lyophilization of red blood cells, red blood cells were lyophilized at different shelf temperature after pre-freeze at -80 degrees C. After lyophilization, the samples were quickly rehydrated by 37 degrees C rehydration solution. The results showed the recovery rate of red blood cells and hemoglobin after pre-freeze at different temperature and lyophilization were > 85% and > 75%, there was not significant difference among these groups, but the concentration of free hemoglobin in -196 degrees C group was significantly higher than that in other groups (P < 0.01). With decreasing of shelf temperature, the lyophilizing time was also prolonged. When shelf temperature was > or = -25 degrees C, samples were not fully lyophilized; when shelf temperature was < or = -30 degrees C, the recovery rate of red blood cells and hemoglobin after lyophilization and rehydration were above 90%; after washed to isotonic state, the recovery rate of hemoglobin of the four groups was similar to each other. In conclusion, only when pre-freezing temperature is between -20 and -80 degrees C and the lyophilizer shelf temperature is < or = -30 degrees C, the effect of lyophilization is better, but the effect of excessively low pre-freezing temperature may even be worse.
Blood Preservation ; Erythrocytes ; cytology ; Freeze Drying ; Hemoglobins ; analysis ; Humans ; Temperature

Neuroglobin (Ngb) is a respiratory protein that is preferentially expressed in brain of mouse and man. In this article, Tibetan antelope, living at altitude of 3 000-5 000 m for millions of years, was selected as the model of hypoxia-tolerant adaptation species. Using reverse transcription polymerase chain reaction (RT-PCR) and Western blot techniques, expression of Ngb gene was amplified and analyzed in antelope brain tissue. Our results showed that Ngb homology protein in Tibetan antelope was identified with more sequence similarity with cattle (96%), sheep (95%), and human (95%). We detected that there were some mutations occurred in the Open Reading Frame of Ngb in Tibetan antelope compared with sheep. Phylogenetic analysis of Ngb chain showed that it was closer to cattle than the others. This study suggests possible roles of central nervous system enriched Ngb in adaptation of Tibetan antelope to extremely high altitude.
Acclimatization ; genetics ; Altitude ; Animals ; Antelopes ; genetics ; Cattle ; Globins ; genetics ; Humans ; Hypoxia ; genetics ; Mice ; Nerve Tissue Proteins ; genetics ; Phylogeny ; Sheep

Energy metabolism plays an important role in life survival for species living in high altitude hypoxia condition. Air-breathing organisms require oxygen to create energy. Tibetans are the well-adapted highlanders in Qinghai-Tibetan Plateau. It was thought that different metabolic approaches could lead to different adaptation traits to high altitude hypoxia. Recently identified hypoxia inducible factors pathway regulators, endothelial PAS domain protein1 (EPAS1)/HIF-2a and PPARA, were involved in decreasing hemoglobin concentrations in Tibetans. Because EPAS1 and PPARA also modulated the energy metabolism during hypoxia, we hypothesized that positive selected EPAS1 and PPARA genes were also involved in unique energy metabolisms in Tibetans. In this brief review, we take a look into genetic determinations to energy metabolisms for hypoxia adaptations traits in Tibetans and mal-adaptive conditions such as high altitude diseases.
Acclimatization ; genetics ; Altitude ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Energy Metabolism ; Hemoglobins ; analysis ; Humans ; Hypoxia ; metabolism ; Oxygen ; metabolism ; Phenotype ; Tibet