1.Use of serum complement C5a as a predictive indicator of fiver injury severity in traumatic rats with hemorrhagic shock
Zhiling ZHAO ; Shuhua CAO ; Yongqiang WANG ; Xinnv XU
Chinese Journal of Emergency Medicine 2011;20(5):498-501
Objective To discuss the feasibility of using serum complement C5a and C5b-9 as predictive indicators of liver injury severity in traumatic rats with hemorrhagic shock.Method Fifry healthy male Wistar rats were randomly(random number)divided into normal group,model 1 hour group,model 3 hours group,model 6 hours group,and model 24 hours group.Plasma CH50,C5a and C5b-9 were detected by using enzyme-linked immunosorbent assay,and rate method was used for determination of plasma aspartate aminotransferase.Paraffin sections of hepatic tissues were used to observe the damage of liver.Results In the model l h group,the CH50 increased significantly and reached the highest value,it began to decline in 3 hours group,and it reached the lowest point in 24 hours group.Compared with the model 3 hours group,6 hours group,and 24 hours group,the level of CH50 in model 1 hour group increased more significantly(respectively P<0.05).A small amount of C5b-9 in the normal group was detected.In the model 1 h group,C5b-9 increased significantly and reach the peak compared with 3hours group,6hours group and 24 hours group,respectively(P<0.05),but in the model 3hours,it began to decline,and in 24 hours group,it reduced to minimum.C5a increased insignificantly in the model 3 hours group,6 hours group and 24 hours group,and peaked in 24 hours group compared with normal group(P<0.05).Aspartate aminotransferase in the model 1 hour group increased significantly and peaked in 24 hours group compared with other groups(P<0.05).Conclusions A large number of complements are activated in the seRing of hemorrhagie shock.C5b-9 and CH50 increase significantly in the early stage,and C5a.increases significantly in the later stage.C5b-9 can be considered as,an initiative factor of liver injury.The low levels of C5a in the early stage may be a mechanism of self-protection of the body.The high levels of CSa in the later stage may be a kind of decompensation,and C5a can be used as a late predictor of disease severity.
2.The effect of TLR4 expression in platelets and activation of platelets on the pathogenesis of thrombocytopenia induced by lipopolysaccharide in mice
Bing WANG ; Yongqiang WANG ; Lei SHAO ; Shuhua CAO
Chinese Journal of Emergency Medicine 2011;20(12):1290-1294
Objective To determine the role of the platelet activation and the expression of platelet Toll-like receptor 4 (TLR4) in thrombocytopenia induced by lipopolysaccharide (LPS) in mice.Methods ICR mice were randomly (random number) divided into control group,model 3,6,12,24,48,72h groups and neutrocytopenia syndrome (NEP) group.The blood samples of mice in model groups were detected 3,6,12,24,48,and 72 hours after intravenous injection of LPS respectively.Anti- neutrophil monoclonal antibody was administered in the mice of NEP group 24 h before injection of LPS,and blood samples were collected 24 h after injection of LPS.The determination of platelet count (PC),mean platelet volume (MPV) and platelet distribution width (PDW) was carried out with full automatic hemocyte analyzer.The plasma tumor necrosis factor- α (TNF- α),macrophage colony stimulating factor (M-CSF) and soluble CD40 ligand (sCD40L) levels were measured by using ELISA.The rate of platelet TLR4 expression was detected by flow cytometry.Comparison between groups was carried out by using ANOVA statistical methods.Results PC was reduced by 30% 3 h after intravenous injection of LPS,and reached the lowest level within 24 h (P <0.01 ).MPV and PDW were increased compared with healthy controls (P <0.01 ).Plasma TNF - α,M - CSF and sCD40L were significantly increased after LPS stimulation (P <0.01 ),and reached the peak during 6 ~ 24 h after LPS administration.PC had correlation with plasma levels of M - CSF ( r =- 0.746) and sCD40L ( r =- 0.573 ).The platelet TLR4 expression was significantly increased 6 h after LPS stimulation.The platelet TLR4 expression in NEP group was significantly lower than that in 24 h model group ( P < 0.01 ).Conclusions The excessive activation platelet represented by increase in sCD40L,MPV,PDW and high levels of M-CSF in macrophages along with platelet TLR4 expression participate in the pathogenesis of thrombocytopenia.The up - regulation of the platelet TLR4 expression is dependent on neutrophils in case of thrombocytopenia induced by LPS.
3.Hpyerglycemic Effect of Arctiin in db/db Mice with Spontaneous Diabetes
Bo ZHANG ; Bing WANG ; Yongqiang WANG ; Shuhua CAO
China Pharmacist 2014;(11):1796-1799
Objective:To investigate the hpyerglycemic action of arctiin in db/db mice with spontaneous diabetes and the underly-ing mechanism. Methods:Totally 40 db/db mice were randomly divided into five groups: the model control group, arctiin group re-spectively with the dose of 75, 150, 300 mg·kg-1 , 300 mg·kg-1 metformin group. The age-matched db/m mice were selected as the normal control group. The mice were administered with corresponding drugs or solvent by gavage for 4 weeks. The oral glucose tol-erance test was carried out at the end of the 3rd week. After the 4-week treatment, all the mice were fasted overnight (12h), and then the body weight and fasting blood glucose ( FBG ) were determined. The concentration of insulin ( INS ) , glycated serum protein ( GSP) , triglyceride ( TG) , total cholesterol ( TC) and adiponection ( APN) were detected. Results:Arctiin could significantly lower the body weight and FBG, improve the glucose tolerance, decrease the serum concentration of INS, GSP, TG, TC and APN(P<0. 05 or 0. 01). Conclusion:Arctiin has benefit effects against glucose/lipid metabolism disorder and insulin resistance in db/db diabetic mice. The mechanism may be related to up-regulating the expression of adiponection.
4.Relationship Between Endotoxin and Prognosis of Multiple Organ Dysfunction Syndrome
Huiyun ZHANG ; Lei ZHANG ; Yongqiang WANG ; Shuhua CAO
Chinese Journal of Nosocomiology 2006;0(05):-
OBJECTIVE To explore the effect of plasma endotoxin on the prognosis of multiple organ dysfunction syndrome(MODS).METHODS Retrospective analysis was carried out on 60 patients of MODS.They were 23 patients in survival group and 37 patients in death group,compared the level of plasma endotoxin on the dd 1,3,5,and 7 after admitted in both groups,and analyzed the relationship between the plasma endotoxin and lethal outcome.RESULTS The level of plasma endotoxin in death group was higher than that of survival group on the dd 1,3,5,and 7(P
5.Bacterial Infection of MODS Patients in Intensive Care Unit
Wenxiu CHANG ; Yongqiang WANG ; Jian LI ; Yinghong XING ; Shuhua CAO
Chinese Journal of Nosocomiology 2006;0(08):-
OBJECTIVE To comprehend the main pathogens and their drug resistance of multiple organ dysfunction syndrome(MODS) patients in ICU.METHODS We retrospectively analyzed all the bacteria isolated from 40 MODS patients in ICU.RESULTS The number of bacteria strains isolated was 173,92 G-bacteria strains made up 53.18%,60 G+ bacteria strains made up 34.68%,and 21 fungi strains made up 12.14%.The top six were Staphylococcus aureus(23.70%,MRSA was 13.87%),Pseudomonas aeruginosa(14.45%),Acinetobacter baumannii(11.56%),Stenotrophomonas maltophilia(8.67%),Candida tropicalis(8.09%),and Enterococcus faecalis(7.51%).The susceptive rate of S.aureus and Enterococcus to vancomycin was all 100%,the susceptive rate of A.baumannii and Klebsiella pneumoniae to carbapenems was high.64% patients had the multiplicity of infection(MOI) which always linked with long period in ICU,respiratory failure and mechanical ventilation.CONCLUSIONS MODS patients have a high morbility of G+ bacteria,fungi and MOI,most pathogens show multi-resistance to commonly used antibiotics.Strengthening the monitoring of infection and reasonable using antibiotics should be taken.
6.Influence of XueBiJing on Apoptosis,Apoptosis-associated Proteins and Activity of Caspase 3 in Spleen of Sepsis Rats
Wenxiu CHANG ; Yongqiang WANG ; Hongmei GAO ; Shuhua CAO
Chinese Journal of Nosocomiology 1994;0(01):-
OBJECTIVE To observe the influence of XueBiJing on apoptosis,apoptosis-associated proteins and activity of caspase 3 in spleen of sepsis rats.METHODS The sepsis models were produced by cecal ligation and puncture(CLP).Ninety-six Wistar rats were randomly divided into four groups:control group(n=8),sham operation group(n=8),model group(n=40),and XueBiJing treatment group(n=40).TUNEL method was used to assess the apoptosis,the activity of caspase 3 and the percent of CD3+ lymphocyte in spleen were tested,and the expressions of FasL and Bcl-2 in spleen were detected by immunohistochemistry methods.RESULTS The integrated optic density(IOD) of FasL expression in spleen for sham operation group was 4.54?1.41,but in model group was much higher(757.96?188.10 at 48 h) and the IOD of Bcl-2 in sham operation group was 594.05?183.32,but in model group was only 22.71?9.22 at 48 h.There were a few apoptosis appeared in sham operation group(0.87?0.51),the activity of caspase 3 was low(4.34?1.34),and the percent of CD3+ lymphocyte in spleen was 63.49%.At 4 h after the CLP operation,there were more apoptosis appeared in model group(8.88?3.68),the activity of caspase 3 reached to 12.22?3.15,and the percent of CD3+ lymphocyte in spleen decreased to 52.09%.After the intervention of XueBiJing,the expression of FasL reduced and Bax raised.The intervention also lightened the activity of caspase 3 and the apoptosis,and made the percent of CD3+ lymphocyte returned to normal level.CONCLUSIONS The activation of apoptosis pathway makes the apoptosis over-appeared,also induces the immune system out-of-balance.The intervention of XueBiJing can ameliorate the immune paralysis of sepsis by regulating the expression of apoptosis-associated proteins and lighten the apoptosis of lymphocyte.
7.Effect of propofol on stress response and immune balance in patients with severe trauma hemorrhage after fluid resuscitation
Bing WANG ; Yongqiang WANG ; Xiang JI ; Shuhua CAO
Chinese Journal of Emergency Medicine 2012;21(4):401-405
Objective To explore the stress response and immune balance in patients with severe trauma hemorrhage after fluid resuscitation and to clarify the clinical effect of Propofol administered with continuous intravenous infusion pump on it.Methods With prospective,randomized and control analysis,54 patients treated with fluid resuscitation following severe trauma hemorrhage admitted from October 1st,2008 to December 1st,2009 were studied.Another 20 healthy volunteers were enrolled as control group (C group). Patients were randomly divided into:conventional treatment group (R group,n =27 ) and conventional therapy combined with propofol treatment ( P group,n =27) as per gender,age,ISS score,estimated blood loss four factors and the principle ofminimum distribution imbalance index.HR,MAP,the levels of stress hormones [ norepinephrine (NE),cortisol (Cor) ],immune function ( T lymphocyte subsets Th1/Th2) and other biomarkers were observed.Mortality within 28 d between R group and P group was compared.ANOVA was used for the comparison of biomarkers,and independent samples t test was employed to compare variables between the two groups. Results Plasma cortisol (Cor),norepinephrine (NE),alanine aminotransferase (ALT),creatinine ( Cr),blood glucose,and Th1 and Th2 lymphocytes in patients were significantly higher than those in healthy volunteers of group C (P <0.01 ),and Th1/Th2 was significantly lower than those in healthy volunteers of C group (P =0.001 ).The ALT (48 h),Glu (6 h),NE (24 h and 48 h) and Cor (6 h and 24 h) of patients in P group were lower than those in R group ( P <0.05 ),and Th1 and Th2 were lower than those in R group at different intervals ( Th1:24h P =0.028,48 hP=0.002 ; Th2:6 h P=0.033,24 h P=0.007,48 h P=0.009),and Th1/Th2 ratio (48 h) was significantly higher in P group than that in R group (P =0.028),but there was no significant difference in mortality within 28d between the two groups. Conclusions Strong stress response,immune suppression and organ dysfunction can occur in the early stages of severe trauma hemorrhage after fluid resuscitation.Propofol can inhibit excessive stress response,modulate the immune balance and protect the important organs in those exsanguination patients from severe trauma.
8.The effect of Xuebijing on the activation-induced cell death (AICD) of T lymphocyte
Wenxiu CHANG ; Shuhua CAO ; Hongmei GAO ; Yongqiang WANG
Chinese Journal of Microbiology and Immunology 2009;29(11):965-970
Objective To observe the influence of Xuebijing injection on apoptosis, apoptotic related gene mRNA levels and activity of caspase3 in activated T lymphocyte. Methods The T lymphocytes were obtained from the spleens of BALB/c mice and be induced to be activated and apoptotic by cultured with Con A + IL-2. Apoptosis was investigated by flow cytometry. RT-PCR was used to detect the expression of Fas, FasL, Bcl-2, Bax, IL-2 mRNA, and the activity of caspase3 in T lymphocyte was also detected by spectrophotometric method. In the mean time, the effect of Xuebijing injection on those parameters was observed. Results After the induction, T lymphocyte apoptosis raised at 18 h. At 6 h after the induction, there was no expression of FasL, Bax mRNA, and no change in the expression of Fas and Bcl-2 mRNA. At 18 h, the expressions of Fas, FasL, Bax mRNA rised and the expression of Bcl-2 mRNA lessened. The activity of caspase3 also ascended. Xuebijing injection can cut down the apoptosis induced by induction, make the expression of Fas, FasL, Bax mRNA decreased and Bcl-2 mRNA improved. The activity of caspase3 also fallen after the Xuebijing injection treated. It can promote the expressions of IL-2 mRNA at early phase of AICD (6 h) and depress the expressions at the late period (18 h). Conclusion The apoptosis of T lymphocyte induced by activation was regulated by the change of Fas, FasL, Bcl-2, Bax mRNA expression. Xuebijing injection can ameliorate the apoptosis through regulating the expression of IL-2 and apoptotic related gene mRNA, improve the proliferation activity of T lymphocyte.
9.Effect of Sishen Pill on NF-κB p65 mRNA and Protein Expression of Colonic Mucosa in Rats with Experimental Ulcerative Colitis
Yan WANG ; Xiangdong ZHU ; Yongqiang DUAN ; Yanfei CAO
Chinese Journal of Information on Traditional Chinese Medicine 2014;(2):49-52
Objective To observe the influence of Sishen Pill on the NF-κB p65 mRNA and protein expressions of colonic mucosa in rats with experimental ulcerative colitis (UC), and identify its underlying mechanism of action. Methods The experimental rats were divided into blank group, model group, Sishen Pill group and SASP group. The models were prepared by TNBS/ethanol enema. Sishen Pill group was intragastrically administrated by Sishen Pill extract 5 g/kg, SASP group by SASP 0.3 g/kg, and blank group and model group by equal volume of normal saline. The morphological injury of colonic mucosa was observed and scored with the naked eyes, and NF-κB p65 gene and protein expression were detected by RT-PCR and immunohistochemical method. Results Inflammation and ulceration on the colonic mucous membrane were found in the model group by naked eyes, and had significant difference with the blank group (P<0.05). The relative expression amount of NF-κB p65 gene and protein of colonic tissues were increased in the model group compared with the blank group (P<0.01). Compared with the model group, the relative expression amount of NF-κB p65 gene and protein in Sishen Pill group were decreased (P<0.05, P<0.01). Conclusion Sishen Pill has effect for treating UC, which is probably related to the activation of NF-κB signal transduction pathway.
10.A study on protective effects of epigallocatechin gallate on encephaledema following traumatic brain injury in rats
Bo ZHANG ; Bing WANG ; Shuhua CAO ; Yongqiang WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;(4):402-407
Objective To observe the inhibitory effect of epigallocatechin gallate (EGCG) on encephaledema following traumatic brain injury (TBI) in rats and its mechanism.Methods 200 Wistar rats were randomly divided into three groups: sham operation (n= 20), model (n= 90) and EGCG (n= 90) groups. The classic Feeney free fall drop method was used to establish the model of TBI. In EGCG group, intraperitoneal injection of EGCG in normal saline 100 mg/kg (10 mL/kg) was immediately given to the rats after model establishment, and in model group, equal amount of normal saline was administered with the same method, once 24 hours for 2 days in all the groups. At 24, 48, and 72 hours after the administration in various groups, the changes of water content, superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in brain tissues were determined, cerebral vascular permeability was evaluated by evans blue (EB) content in the brain tissues, the changes of expressions of aquaporin-4 (AQP4) and glial fibrillary acidic protein (GFAP) in brain tissues were determined by immunohistochemical and Western Blot, and the cerebral histopathological changes were observed in various groups.Results Compared with sham operation group, the water content, the vascular permeability and MDA level in brain tissues were significantly higher, while the cerebral SOD activity was significant lower in the model group; the scores of cells with positive AQP4 and GFAP expressions (IHC score) were obviously increased at 24 hours and 72 hours after model establishment, and the levels of expressions of AQP4 and GFAPprotein [integral absorbance (IA) value] were markedly enhanced in model group than those in the sham operation group, the changes being more remarkable at 72 hours after model formation [water content in brain tissues: (89.71±0.94)% vs. (78.34±0.87)%, EB content (μg/g): 9.13±0.66 vs. 2.71±0.72, SOD activity (U/mg): 63.53±12.57 vs. 130.85±9.91, MDA (nmol/mg): 10.19±1.47 vs. 4.57±0.74, IHC score of AQP4: 8.81±1.75 vs. 2.76±0.82, IHC score of GFAP: 9.47±1.32 vs. 6.71±0.52, expression of AQP4 protein (IA value): 1.53±0.05 vs. 0.42±0.05, expression of GFAP protein (IA value): 1.45±0.05 vs. 0.62±0.04, allP < 0.01]. Compared with the model group, the cerebral water content, MDA, IHC scores and protein expressions of AQP4 and GFAP, and cerebral vascular permeability were significantly decreased, while the SOD activity was obviously increased in the EGCG group, and the changes being more significant at 72 hours after model establishment [water content of brain tissues: (86.59±0.89)%, EB content (μg/g): 7.82±0.32, SOD activity (U/mg): 107.58±10.87, MDA (nmol/mg): 5.61±1.64, IHC score of AQP4: 6.92±0.71, IHC score of GFAP: 6.71±0.52, expression of AQP4 protein (IA value): 1.14±0.06, expression of GFAP protein (IA value): 1.21±0.07, all P < 0.01]. Imunohistochemical assay showed: the cerebral contents of AQP4 and GFAP positive cells in the rats of EGCG group were decreased, and their color became lighter.Conclusion The inhibition of EGCG on encephaledema following TBI in rats is related to its effects of decreasing the cerebral vascular permeability, enhancing the level of SOD activity, depressing MDA level and the expressions of AQP4 and GFAP.