ABSTRACT
Objective:To investigate the effect of sodium bicarbonated Ringer's solution on fluid resuscitation in rabbits with traumatic hemorrhagic shock.Methods:The rabbit model of traumatic hemorrhagic shock was established by Lamson's method. All 30 Japanese white rabbits were randomly divided into three groups: the normal control group (NC group, n=10), the sodium lactate Ringer's solution group (LRS group, n=10), and the sodium bicarbonate Ringer's solution group (BRS group, n=10). In the resuscitation period, rabbits in the NC group received only carotid artery and femoral artery intubation and systemic heparinization, without beating or bleeding. Rabbits in the LRS group received all blood loss and the same amount of sodium lactate Ringer's solution through the ear vein, and rabbits in the BRS group received all blood loss and the same amount of sodium bicarbonate Ringer's solution for resuscitation. Mean arterial pressure (MAP) and heart rate (HR) were recorded at each time point before shock, namely basal stage (T1), end of shock (T2), end of resuscitation (T3) and 2 h after resuscitation (T4), and arterial blood was collected for blood gas analysis. The death rates of the experimental animals in each group were recorded at 12, 24, 36 and 48 h, respectively, and the survival rates were calculated. One-way analysis of variance was used for comparison between groups, LSD method was used for multi-group comparison, and Kaplan survival analysis was used for comparison of survival time of each group. Results:There were no significant differences in age, body mass index and basal blood pressure among the three groups ( P>0.05). After 2 h of fluid resuscitation, the improvement effect of the BRS group was significantly better than that of the LRS group (MAP: 87.79±2.94 mmHg vs 79.24±3.81 mmHg; HR: 191.9±16.8 times/min vs 211.3±15.6 times/min; lactic acid: 6.09±1.94 mmol/L vs 7.89±2.47 mmol/L; arterial blood pH: 7.31±0.04 vs 7.21±0.04, all P<0.05). The survival rates of experimental animals in the BRS group at 12, 24, 36 and 48 h were significantly higher than those of the LRS groups (90% vs 80%, 80% vs 60%, 70% vs 50%, 60% vs 30%; all P<0.05), and the survival time of the BRS group was longer than that of the LRS group. Conclusions:During the fluid resuscitation of rabbits with traumatic hemorrhagic shock, sodium bicarbonate Ringer's solution can significantly reduce the blood lactic acid level, maintain the acid-base balance and hemodynamic stability, and improve the survival rate of rabbits with traumatic hemorrhagic shock.
ABSTRACT
Objective To compare the accuracy of three classification systems [determinant based classification (DBC),Revision of the Atlanta classification (RAC),and Atlanta classification (AC)] to stratify severity of acute pancreatitis (AP),and to analyze the association between different severity categories and clinical outcomes.Methods In this retrospective study,we reviewed the clinical data of 458 patients with AP admitted to our unit from January 2015 to December 2017.AP severity was stratified according to the three classification systems (DBC,RAC,and AC) respectively.The classification accuracy of three classification systems was analyzed.Receiver operating characteristic analysis (area under the curve) compared the accuracy of each classification.Multi-factors logistic regression analysis identified the independent risk fators for mortality of AP.Results Among the three classification systems,there were significant differences in the mortality rate,invasive treatment rate,ICU monitoring rate and the average hospitalization time among the three subtypes (P＜0.001).The RAC and DBC were comparable,but performed better than AC in predicting mortality (AUC 0.94 and 0.95 vs.0.63,P＜0.001),ICU admission (AUC 0.90 and 0.88 vs 0.60,P＜0.001).The DBC performed better than the RAC and OAC in predicting the need for intervention (AUC 0.88 vs 0.69 and 0.68,P＜0.001).Persistent organ failure (OR=13.131,P=0.003) and infected necrosis(OR=9.424,P=0.014) were independent risk factors for mortality.Conclusion The accuracy of DBC and RAC to stratify the severity of AP was significantly higher than that of AC.The accuracy of DBC in predicting clinical outcome was genarally higher than that of RAC and AC.Infectious necrosis and persistent organ failure were the independent risk fators for mortality.
ABSTRACT
Objective To study the system of streptococcal C5a peptidase (ScpB) and the specif-ic antibody levels in serum, lung, vagina and recta after subcutaneous and intranasal immunization with dif-ferent doses of C5a peptide. Methods Recombinant protein C5a peptide was expressed in E. coli strain BL21 and purified by affinity chromatography. The expressed product was identified by SDS-PAGE and pep-tide mass fingerprinting (PMF). BALB/c mice were subcutaneously and intranasally injected with different doses of ScpB. Antibody titer was tested by ELISA. Opsonophagocytosis assay was used to test the function of antibody. Results ScpB protein was successfully expressed and purified. The probability based mouse score of ScpB was 175 by PMF analysis. ELISA data showed that both subcutaneous and intranasal immtmi-zation could elicit significantly higher levels of IgG in immunized mice serum than that of control group (P ＜0.01), 30 μg group waa better than 5 μg and 10 μg group. Intranasal immunization could elicit higher lev-els of IgA in lung, vagina and rectum (P ＜0.001) while system immunization could not. Opsenophagocyto-sis tests indicated that anti-serum of ScpB had opsenophagocytic activity than that of control (P ＜ 0. 05).Conclusion The results demonstrated that intranasal immunization with ScpB could induce significantly higher levels of lgG and IgA, and its anti-serum had better opsenic activity.
ABSTRACT
AIM: To solve the two difficulties of bone resorption and inflammation in rheumatoid arthritis, clone the recombinant human osteoprotegerin (OPG) and mycobacteria heat shock protein 70 (HSP70) functional gene,and study the expression and activity of OPG-HSP70 fusion protein in E.coli. METHODS: Experiments were performed at the Laboratory of Department of Immunology, Capital Medical University from May 2006 to September 2007. Complementary DNA encoding full length OPG protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma cell line MG63 and cloned into pGEMT-Easy vectors. Then, using the recombinant plasmid as the template,the DNA encoding the fusion protein OPG-HSP 70 was amplified by PCR,and was inserted into prokaryotic expression vector pET-28a. Construct pET-28a-OPG-HSP 70 was used to transform into competent E.coli. BL21(DE3) which were induced by isopropyl B-D-thiogalactopyranoside (IPTG),and the fusion protein from above E.coli was collected. Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blotting were performed to identify OPG-HSP 70 fusion protein. The experiments of osteoclast inhibition and restraining inflammation were used to detect the bioactivity of fusion protein. RESULTS: ①The complementary DNA encoding full length OPG protein was obtained. OPG-HSP 70 fusion gene obtained in this experiment was successfully inserted into pET-28a vector. OPG-HSP70 fusion protein was expressed when transformed into E.coli.BL21(DE3). ②SDS-PAGE indicated that the fusion protein was large expressed at the molecular weight of Mr22 000, but there were no band in the total lysate of bacteria harboring pET-28a-OPG-HSP70 without IPTG induction group. ③Western-blotting indicated that the OPG-HSP70 fusion protein could specifically react with anti-human OPG monoclonal antibodies. ④The osteoclast inhibition test demonstrated that the fusion protein could reduce the number of osteoclast, and had the ability to inhibit bone absorptions in vitro. ⑤The experiment of restraining inflammation showed that the fusion protein could significantly reduce the inflammation of delayed type hypersensitivity (DTH) mice, which explained that HSP70 in fusion protein had the inflammation inhibitory bioactivity. CONCLUSION: OPG-HSP70 fusion protein is expressed in E.coli.BL21(DE3),and function study in vitro illustrates the bioactivity of fusion protein.
ABSTRACT
Objective To investigate the function of human glia maturation factor gamma to induce the proliferation and differentiation of neural stem cells into astrocytes.Methods The neural stem cells were isolated from cerebral cortex of 11-to 12-day embryoes of SD rats.After three passages,the cells could react with nestin antigen,which demonstrated that the cells were neural stem cells.The neural stem cells were divided into 4 groups,control group,FBS group FBS,GMFG 5 ?g/L group and 5 ?g/L of GMFG,GMFG 10 ?g/L group with medium and 10 ?g/L of GMFG.Results In the control group,the cells had no significant differentiation in two days and slight branches were observed at the fourth day.In FBS group,the cells differentiation into astrocytes within three days,which grew on the surface of the culture flasks.In GMFG group,the cells differentiated into astrocytes in one day,and the differentiation of cells from the 10 ?g/L group was more obvious than that of cells from the 5 ?g/L group.The cells from the 10 ?g/L group reacted weakly with GFAP antigen.Conclusion GMFG can induce proliferation and differentiation of neural stem cells into astrocytes effectively in vitro.
ABSTRACT
Objective To investigate the inhibitive effect of antisense VEGF165 cDNA on growth and angiogensis of human neuroblastoma in nude mouse models.Methods Three kinds of SH-SY5Y cells which had been stably transfected by sense VEGF165 cDNA,antisense VEGF165 cDNA and empty vector respectively,were inoculated subcutaneously to nude mice.The size of subcutaneous tumors was measured,and the morphology of tumor tissue was observed by microscope.The microvessel density(MVD)in tumor mass was analyzed by CD31 immunohistochemistry staining.Apoptotic cells were detected by the electron microscope.Results The growth of tumor mass,which was inoculated with antisense VEGF,significantly decreased in nude mice compared with empty vector group(P