Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

Aim To examine the effect of different dosages hydroxyethyl starch(HES)130/0.4 on intercellular adhesion molecule 1(ICAM-1)expression in lung tissue of acute lung injury in endotoxemic rats and explore the role of MAPKs pathway in its expression.Methods Thirty six healthy Sprague Dawley(SD)rats weighing 270~320 g were randomly divided into 6 groups with 6 animals in each group.In group H1-H4,1 min after lipopolysaccharide(LPS)5 mg?kg-1 intravenously administration,HES 130/0.4 with 3.75,7.5,15,30 ml?kg-1 were infused intravenously respectively at a rate of 0.2 ml?min-1.In group L,saline instead of HES 130/0.4 was administered.Group N served as control by giving the same volume of saline.The animals were anesthetized with pentobarbital.Right external jugular vein was injected with LPS.The animals were killed 4 hours after LPS injection for determination of total protein,WBC,MPO,W/D,ICAM-1 protein and mRNA and MAPKs activity.Results Compared with control group,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were increased significantly in group L.Compared with group L,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were decreased significantly in group H1 and H2,especially at the dosage of 7.5 ml?kg-1.Conclusion HES 130/0.4(7.5 ml?kg-1)can attenuate inflammatory response of acute lung injury induced by LPS,which may be related with inhibiting the expression of ICAM-1 protein and mRNA through MAPKs signal pathway.

Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.

Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.

AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.

OBJECTIVE:Aspirin sustained release capsules(A-SRC) were prepared The release rate and bioavailability of A-SRC were studies METHODS:To study release mechanism by determining dissolution rate and to study relative bioavailability of A-SRC by determining serum concentrations in rabbits RESULTS:The release profile of A-SRC fitted zero-order kinetics This sustained release preparation had good release stability and high bioavailability in comparison with commonly-used aspirin sustained release capsules CONCLUSION:A-SRC showed obviously sustained release effect The drug concentration in blood kept steady and lasted long after oral administration,therefore the times of administration could be decreased Because the light-excipients were used in A-SRC,which had floating properties and could make drug release slowly A-SRC could decrease the irritations to gastrointestinal tract and other side effects

Objective: To evaluate the testing level of the relevant items in food inspection agency laboratory objectively, analyze the existing problems and help them enhance the ability of detection.Methods: The blind sample assessment of sodium saccharin in drinks was organized and performed.The test samples at high and low concentrations were prepared and studied by the tests of homogeneity and stability.Those met the requirements of blind sample assessment were randomly distributed to 193 laboratories and the returned data were analyzed statistically.Results: Totally 182 valid data were collected.Among the reported results, those from 85 laboratories were satisfied in the low concentration group with the satisfaction rate of 87.6% , those from 12 laboratories were not satisfied, and no results were suspicious;as for the high concentration group, the above data was 67(78.8%), 12 and 6, respectively.The overall satisfaction rate was 83.5%.The results, especially the dissatisfied and suspicious data, were analyzed by the original records of each participating laboratory, the analysis focused on 8 aspects including detection method, instrument brand, control product variety, pretreatment method, recovery rate, column selection and the other influencing factors, and suggestions were given to every laboratory to improve its own situation.Conclusion: The statistical data reflect that our food testing laboratory has strong detection ability in the determination of saccharin sodium in beverages, and can provide powerful technical support for the regulatory authorities.

0.05).The concentration of urinary KYNA,metabolite of the KYN,was significantly lower in SHRs compared to WKYs(7.8?1.8 vs 19.9?3.5 ?mol/24 h P=0.013).Both KAT activity in renal cortex and KYNA content in urine were negatively correlated to blood pressure(r=-0.418,P=0.023;r=-0.723,P=0.001).Conclusion The declined activity of KAT in renal cortex and the deficiency of KYNA concentration in urinary may affect blood pressure regulation in SHR by renal metabolite of the KYN.

To improve the effectiveness of nurse-patient communication and build a harmonious nurse-patient relationship, a list of melodrama of the whole nursing process from admission to discharge was designed and demonstrated. This means of training by scenarios demonstration significantly enhanced nurses' awareness and skills of nurse-patient communication with high recognition.

OBJECTIVETo investigate the effect of allogene hepatic nonparenchymal cell (NPC) on the survival of grafted skin in mice and its underlying mechanism.

METHODSSixty-five C(3)H and fifty-eight C(57)BL/6 mice were employed in the study. Twenty C(3)H mice were used as skin donor and forty as the source of hepatic NPC. The rest five served as the stimulators of mixed lymphocyte culture (MLC) before and on the 7th, 18th, 30th, and 60th day after NPC infusion with 1 at each time point. MLC was determined and expressed as count per minute (CPM). Fifty-eight C(57)BL/6 mice were further divided into experimental (E, n = 50) and control groups (C, n = 8). The mice in C group only underwent skin grafting without NPC infusion. The mice in E group received with 2 x 10(7) NPC via caudal vein, followed by peritoneal injection of cytoxan (200 mg/kg) 48 hours later; They were grafted with skin donated from C(3)H mice 18 days after injections. The survival time of the mice in the two groups was observed. The serum levels of interleukin-4, chimera and MLC in the two groups were determined before and on 7th, 18th, 30th, 60th days after NPC infusion, and micro-chimera were aslo assessed on the 1st and 3rd day after NPC infusion. Five mice were sacrificed at each time point.

RESULTSThe survival time of skin graft in E group (70.0 +/- 17.2 day) was obviously longer than that in C group. The serum levels of IL-4, chimera in E group were increased gradually, while MLC response decreased gradually. The serum IL-4 level reached 251.5 +/- 11.0 ng/L and splenic chimera level to 26.30 +/- 1.04% on the 60th day after NPC infusion.

CONCLUSIONThe high levels of IL-4 and chimera might play important roles in inducing and maintaining immune tolerance.

Animals ; Female ; Graft Survival ; immunology ; Hepatocytes ; immunology ; Immune Tolerance ; Interleukin-4 ; metabolism ; Male ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Skin Transplantation ; immunology ; Surgical Flaps ; Transplantation Chimera ; Transplantation, Homologous ; immunology