Objective To explore the ideal resuscitation pressure for uncontrolled hemorrhagic shock in different ages of rats.Methods Ninety-six rats were divided into 3 groups by ages (6 weeks,16 weeks and 12 months representing pre-adult,adult and elderly rats,respectively).Uncontrolled hemorrhagic-shock model was established in all rats.Then every group was divided into 4 groups by the different target pressures (untreated group,40 ～ 50 mmHg group,＞ 50 ～ 60 mmHg group,and ＞ 60 ～ 70 mmHg group).We recorded the survival time and 24 h survival rate of rats in each group and compared the blood gas analysis,blood lactate and hematocrit (Hct) level in each group at 90 min of establishing model.Results Different ages had different optimal recovery pressure.The optimal target resuscitation pressures for 6 weeks,16 weeks and 12 months rats were 40 ～50 mmHg,＞ 50 ～ 60 mmHg and ＞ 60 ～ 70 mmHg,respectively.Under the ideal pressure they had a better survival rate and lower blood lactate level (P ＜ 0.01 or P ＜ 0.05).Conclusious Hemorrhagic-shock rats at different ages have different ideal target resuscitation pressures during active hemorrhage.

Objective: To analyze the influencing factors of whole blood colloid osmotic pressure (COP) and predict reference range of plasma protein for safe COP to guide clinical infusion of protein in critically ill patients. Methods: Physical data and blood gas analysis of 405 patients were collected. hTe patients were divided into 2 groups by COP: group A (COP≤18 mmHg) and group B (COP>18 mmHg). The serum proteins including total protein (TP), albumin (Alb), globumin (Glb) and ifbrinogen (FIB) were detected. Results: APACHE II of group B was signiifcantly lower than that of group A (P<0.05). hTe survival rate, TP, Alb, Glb and FIB of group B were significantly higher than these of group A (P<0.05).Standardized regression coeffcient of Alb, Glb and FIB was 0.518,0.283 and 0.113 (P<0.05); the 95% reference range of 4 types of protein level in group B: Alb>23.3 g/L,Glb 12.6-37.6 g/L and FIB 1.3-8.7 g/L; 5 reformed equations were made. Conclusion: The main influencing factors of COP include Alb, Glb and FIB. We can use the reference range of 4 types of protein level to guide the clinical management of protein agents, and reformed equations can be used to preliminarily forecast COP in critically ill patients.

Objective To study the changes of S100βin serum and brain tissue of rats with sepsisassociated encephalopathy (SAE).Methods After placement of pole plates of electroencephalogram (EEG) on the brain cortex,thirty SD rats were randomly (random number) divided into three groups post hoc:sepsis group in which rats were made to be sepsis models by cecal ligation and puncture (n =18),normal group (n =6) and sham operation group (n =6).The sepsis-associated encephalopathy was diagnosed with electroencephalogram taken by RM6240 physiological signal recorder.Meanwhile,heart rate and blood pressure were recorded.Eight hours later,serum and brain tissue of sacrificed rats were taken for measuring S100β.The results ware analyzed with one-way ANOVA.Results Of 18 sepsis rats,3 were dead,8 without SAE and 7 with SAE.The levels of S100βin serum and brain tissue of rats with SAE group were significantly higher than those in normal group and sham operation proup (P ＜ 0.05).The Ratio of brain/serum S100βin rats with SAE group was higher than that in rats without SAE (1.74 vs 1.51,P ＜ 0.05).Conclusions Significantly high level of serum S100βwas a reliable bio-marker for diagnosis of SAE in rats.

Objective To investigate the effect of continuous veno-venous hemofiltration (CVVH) on the peripheral blood levels of high mobility group box chromosomal protein 1 (HMGB-1) in patients with systemic inflammatory response syndrome (SIRS) or sepsis.Methods Thirty patients with SIRS or sepsis in Intensive Care Unit of our hospital were scheduled for treatment of CVVH.The replacement liquid was put in by the pattern of pre-dilution of 40％ and post-dilution of 60％.The flow-rate was 3 L/h.The blood flow-rate was from 200 to 300 ml/min.5 ml blood from right radial artery was got at the time points of preCVVH,CVVH 2 h,6 h,8 h and post-CVVH 12 h and the serum was stored at the temperature of-20 ℃ after high speed centrifugation,and 2 ml filter liquor was reserved at the time point of CVVH 6 h.The concentration of serum and filter liquor HMGB-1 was measured by ELISA,but that of TNF-α and IL-6 were measured by radioimmunity.Results 30 patients adept CVVH therapy within 24 h to the hospital,and their therapy time was 2 ～ 5(2.4 ± 1.5) d.Among them,17 cases survived and 13 cases died with a fatality rate of 43.3％.Serum concentration of HMGB-1 decreased from the baseline in patients,although this decrease was not statistically significant[(11.88 ± 6.06) ng/ml,(11.97 ± 5.66) ng/ml,(11.94 ± 5.94) ng/ml,(11.73 ± 5.19) ng/ml vs (13.87 ± 4.68) ng/ml,P ＞ 0.05],Serum concentration of both TNF-αand IL-6 after therapy significantly decreased compared to the baseline in patients [TNF-α: (0.28 ± 0.15)ng/ml,(0.30 ± 0.14) ng/ml,(0.29 ± 0.19) ng/ml,(0.33 ± 0.19) ng/ml vs (0.41 ± 0.12) ng/ml,IL-6:(408.20 ±92.18)pg/ml,(250.51 ± 107.34)pg/ml,(276.00 ± 126.20) pg/ml,(315.16 ± 130.97) pg/ml vs (513.35 ± 125.95) pg/ml,P ＜ 0.05].Conclusions CVVH could decrease the concentration of HMGB-1 in peripheral blood,which would be one of the mechanisms of action for CVVH on sepsis.

Objective To investigate the effect of Xuebijing injectiong on lipopolysaccharide(LPS)-induced apoptosis of CD4+CD25+regulatory T cells(Tregs)and immune function of effector T cells(Teff)in vitro.Method CD4+CD25+Tregs isolated from rat spleens were divided into the control group,anti-CD3/CD28 group,anti-CD3/CD28+LPS group,anti-CD3/CD28+Xuebijing injection group,and anti-CD3/CD28+LPS+Xuebijing injection group.The apoptosis rate and expression of forkhead/winged helix transcription factor p3 (Foxp3)and cytotoxic T-lymphocyte-associated antigen 4(CTLA-4)of CD4+CD25+Tregs were detected by flow cytometry(FCM),and the secretion of IL-10 of Tregs was measured by ELBA on day 3.CD4+CD25-T cells were co-cultured with CD4+CD25+Tregs(1:1)for 68 hours,proliferative activity of Teff was determined by MTT,and interleukin(IL)-2/sIL-2Rα levels were measured by ELISA.Results The apoptosis rate of CD4+CD25+Tregs in anti-CD3/CD28 group was 33.70± 3.06%,which was significantly higher than that in control group(12.84±0.84%).Also,apoptosis rate of CD4 CD25+Tregs in anti-CD3/CD28+LPS+Xuebijing injection group(45.13±2.70%)was much higher than that in anti-CD3/CD28+IPS group(29.41 ± 1.63%,P＜0.01).The expression of Foxp3 as well as CTLA-4,and the secretion of IL-10 were markedly decreased along with increases in the apoptosis rates.Compared with control group(54.48%),the mean inhibitory rate of Teff proliferative activity in response to Con A was significantly decreased in anti-CD3/CD28+Xuebijing injection group(31.26%,P＜0.05),and it was markedly decreased in anti-CD3/CD28+LPS+Xuebijing injection group comaped to anti-CD3/CD28+LPS group(P＜0.01).In addition,IL-2 levels in the supernatant of anti-CD3/CD28+Xuebijing injection group and anti-CD3/CD28+LPS+Xuebijing injection group were significantly higher than those of anti-CD3/CD28+IPS group(P＜0.01).Conclusions The inhibitory activity of CD4+CD25+Tregs on Teff appears to be upregulated by IPS stimulation in vitro,and Xuebijing injection could markedly enhance apoptosis of CD4+CD25+Tregs,thereby improving suppressive immune function of Teff.

Objective To evaluate the diagnostic and prognostic value of the serum procalcitonin ( PCT ) level in the non-acquired immune deficiency syndrome ( AIDS ) immunocompromised critically ill patients suspected to have infection. Methods A retrospective study was conducted in the non-AIDS immunocompromised patients who were admitted to Department of Critical Care Medicine of Xiangya Hospital, Central South University during January 2011 to December 2014. Demographic characteristics, underlying disease, acute physiology and chronic health evaluationⅡ( APACHEⅡ) score at admission, and clinical records including baseline and peak levels of temperature, white blood count ( WBC ), PCT, and survival rate within 28 days, infection focus, infectious agents ( bacterial, fungi or mixed infection ), and the severity of infection ( sepsis, severe sepsis, or septic shock ) were recorded. Receiver operating characteristic ( ROC ) curve was plotted, and the diagnostic and protective value of above parameters was evaluated. Results A total of 98 patients ( 43 male and 55 female ) were enrolled in the study with a median age of 44 ( 28, 52 ) years old and a median APACHEⅡscore of 17 ( 11, 20 );47 with malignant hematological tumor, 45 with autoimmune diseases, and 6 post solid organ transplantation. Among them 53 patients ( 54.1%) died within 28 days. Twenty-seven patients were diagnosed as systemic inflammatory response syndrome ( SIRS ) without infection. Among 71 patients with infection, 45 were diagnosed as bacterial infection, 10 with fungal infection, and 16 with mixed infection. Sepsis was diagnosed in 7 patients, severe sepsis in 32 patients , and septic shock in 32 patients .①There was no statistical significance in the baseline and peak levels of PCT and WBC, or baseline level of temperature between the groups of SIRS patients without infection and infected patients. The peak level of temperature was significantly higher in the patients with infection as compared with that of the SIRS without infection patients [℃:39.4 ( 38.9, 40.0 ) vs. 38.8 ( 37.8, 39.2 ), Z=-3.268, P=0.001 ]. It was showed by subgroup analysis that in patients with hematological malignant disease or autoimmune diseases, higher level of body temperature was found in infection group compared with non-infection SIRS group [℃:39.5 ( 39.0, 40.0 ) vs. 39.0 ( 38.4, 39.4 ), Z=-2.349, P=0.019;39.0 ( 38.4, 39.5 ) vs. 38.2 ( 37.0, 38.9 ), Z=-2.221, P=0.026 ].②The baseline level of PCT (μg/L ) were 0.54 ( 0.20, 4.19 ), 2.78 ( 0.50, 9.54 ), 1.00 ( 0.45, 6.89 ), and 0.22 ( 0.07, 1.86 ) in non-infection SIRS patients or the patients with bacterial, fungal, and mixed infection, respectively. The peak level of PCT (μg/L ) were 4.19 ( 1.95, 13.42 ), 12.37 ( 3.82, 45.89 ), 1.82 ( 0.49, 17.86 ), and 5.14 ( 2.66, 12.62 ), respectively, in each subgroup. When the comparison was conducted among the patients with different infectious agent, the baseline level of PCT in patients with bacterial infection was significantly higher than that in SIRS patients without infection ( P=0.026 ) and mixed infection patients ( P=0.001 ), and the peak level of PCT was significantly higher than that in the SIRS patients without infection ( P=0.009 ) and the patients with fungal infection ( P=0.016 ). ROC curve showed that the higher value was found in the baseline and peak levels of PCT for diagnosis of septic shock in all patients [ area under ROC curve ( AUC ) of baseline level = 0.681±0.054, P = 0.001; AUC of peak level = 0.690±0.054, P=0.002 ], and the same value was also found in the baseline and peak levels of PCT for diagnosis of bacterial infection in the patients with malignant hematological tumor ( AUC of baseline level=0.687±0.080, P=0.008;AUC of peak level=0.697±0.079, P=0.021 ).③The peak level of PCT (μg/L ) were 4.05 ( 0.53, 31.22 ), 5.78 ( 2.14, 16.68 ), and 11.64 ( 2.94, 58.14 ) in subgroup of patients with sepsis, severe sepsis and septic shock, respectively, and they showed no statistical significance among subgroups ( P>0.05 ). A high serum level of peak PCT strongly indicated the presence of septic shock ( AUC=0.646±0.060, P=0.019 ), especially in the subgroup of patients with systemic autoimmune disease ( AUC=0.689±0.081, P=0.035 ).④The peak level of PCT (μg/L ) in the APACHEⅡ>18 group ( 38 cases ) was significantly higher than that of APACHEⅡ≤18 group [ 60 cases, PCT (μg/L ):11.64 ( 3.36, 39.39 ) vs. 4.42 ( 1.32, 14.70 ), P=0.016 ];there was a certain correlation between the peak level of PCT and the severity of the disease.⑤The peak level of PCT in death group was significantly higher than that of the survival group [μg/L:9.07 ( 3.05, 33.09 ) vs. 4.19 ( 1.26, 14.61 ), P=0.043 ]. ROC curve showed that the peak level of PCT might be valuable in predicting the prognosis in immunocompromised patients ( AUC=0.619±0.057, P=0.043 ). Conclusions The serum level of PCT is found to be a reliable marker for the diagnosis of bacterial infection in immunocompromised critical patients, especially in those with hematologic malignancy. Additionally, PCT provides a useful tool for evaluating the severity of infection and the prognosis of critically ill patients.

Objective To evaluate the correlation between the level of CD4+CD25+regulatory T cells in periphend blood and disease severity in patients with sepsis.Methods Thirty-six septic patients and 5healthy controls were enrolled.Septic patients were divided into sepsis group(n=10),severe sepsis group (n=15)and septic shock group(n=11).The lymphocyte was seperated from peripheral blood and marked by PE-CD4 and FTTC-CD25 monoclonal antibody,the level of CD4+CD25+ regulatory T cells was detected by flow cytometry,and the clinical data of APACHE Ⅱ scores of septic patients was considered in 24 hours.The correlation between level of CD4+CD25+ regulatory T cells in peripheral blood and APACHE Ⅱ scores in septic patients was analyzed.Results Compared with the healthy controls [(5.48±0.98)%],the level of CD4+ CD25+ regulatory T cells in sepsis group(10.31±2.32)%,severe sepsis group(14.27 43.33)%,septic shock group(15.32±3.98)% had a significant increase(P＜0.05 or＜0.01).The log value of regulatory T cells in each group correlated positively with the APACHE Ⅱ scores(r=0.829,P=0.032;r=0.868,P=0.021;r=0.913,P=0.009),and the total coefficient of correlation was 0.903(P=0.013).Conclusion The level of CD4+CD25+ regulatory T cells in peripheral blood in septic patients has an abnormal increase,and their levels are related with the severity of disease.

Objective To investigate the mechanism of growth hormone inhibiting IPS-induced apoptosis of alveolar type Ⅱ epithelial cells in rats. Method Isolated and purified AEC Ⅱ cells of SD rats were divided into 5 groups,8 duplicate wells in each group. Group I served as control group; group Ⅱ:LPS 10 ug/ml;group Ⅲ:LPS 10 ug/ml + GH 50 ng/ml;gronp IV :LPS 10 ug/ml + GH 100ng/ml; group V: LPS 10 ug/ml + GH 200 ng/ml. LPS was finally added into wells in group Ⅱ～V . After the cells were incubated for 24 hours, the apoptosis rate and necrosis rate of AEC Ⅱ cells stained with Annexin V/PI were detected by flow cytometry and Fas protein of AEC Ⅱ cells were measured by immunocytochemistry. Results (1) The apoptosis rate and necrosis rate of AECⅡ cells in group Ⅱ,Ⅲ, Ⅳ and V were significantly hitOer than those in group Ⅰ( qapoptosis rate Ⅰ, Ⅱ =12.26,qnecroeis Ⅰ,Ⅱ=18.34, qapoptosisⅠ.Ⅱ=9.63,qnecrosisⅠ,nⅡ=5.75,qapotosisⅠ,Ⅳ= 9.15,qnecrosisⅠ,Ⅳ= 5.39, qapotosisⅠ,Ⅴ = 10.87, qnecrosisⅠ,Ⅴ = 5.91, P 0.05), but lower in group Ⅲ,IV and V than those in group Ⅱ(qapoptosis Ⅱ,Ⅲ= 15.24, qpecrosisⅡ,Ⅲ=16.38, qapoptosisⅡ.Ⅳ = 15.95,qnecrosisⅡ.Ⅳ=16.95, qapoptosis rate Ⅱ,Ⅴ=14.57, qnecrosisⅡ.Ⅴ = 15.61,P<0.05). (2)The positive rate of Fas expression on AEC Ⅱ cells in group Ⅱ,Ⅲ, Ⅳ and V was obviously higher than that in group Ⅰ. ( q Ⅰ.Ⅱ=35.67, qⅠ ,Ⅲ=14.32, qⅠ,Ⅳ = 13.87, qⅠ.Ⅴ=26.16, P<0.05), but lower in gronpⅢ ,Ⅳ and Ⅴ than that in gronp Ⅱ(qⅡ,Ⅲ=12.54, qⅡ,Ⅳ = 13.02, qⅡ,Ⅴ =6.96, P<0.05). Conclusions GH can probably de-crease the apoptosis of AEC Ⅱ cells by inhibiting Fas expression.

Aim To observe the change of NF-?B and expression of Bax and Bcl-2 related to apoptosis regulation in the cerebral infarction rat and study the protective effect of iridoid glycoside(IG) extracted from cornus officinalis on focal cerebral ischemia injury and its mechanism of action.Methods Rats were pretreated with drugs by mouth for 7 d and a focal cerebral infarction model of rat was induced by photochemical reaction.Immunohistochemistry and western blot were used to detect the change of expression of NF-?B,Bax and Bcl-2 in the cortex.Results Compared with control group,expression of NF-?B and Bax were increased while expression of Bcl-2 was decreased.Compared with model group,IG(IG 20、60、180 mg?kg~(-1)) markedly decreased the expression of NF-?B and Bax and increased expression of Bcl-2.Conclusion IG has therapeutical effect on cerebral infarction through regulating the expression of NF-?B and apoptosis related genes

Objective To establish sepsis-associated encephalopathy (SAE) animal models by using neurobiology score,electroencephalography (EEG),somatosensory evoked potentials in order to provide evidence for early clinical diagnosis of SAE.Methods A total of 30 rats were weighted,numbered,and monitored with EEG electrodes 10 days before modeling.Ten days later,rats were weighted,numbered,and divided randomly (random number) into groups.Rat models of sepsis were made by cecal ligation and puncture (CLP).The changes of their neurological behaviors were observed and EEG was used to monitor at 4,6,8,12 and 24 hours after CLP.The changes of EEG waveform and somatosensory evoked potentials were analyzed and recorded.Rat models of sepsis were divided into sepsis + non-SAE group and SAE group based on the presence or absence of EEG or somatosensory evoked potentials changes ~thin 24 hours.Rats were sacrificed 24 hours later,and histopathological changes of brain tissue were observed under electronic microscopy.Thus,the feasibility of establishing early SAE animal model by monitoring the changes of neurological behaviors,EEG and somatosensory evoked potentials was evaluated.Results SAE could be early diagnosed by using neurobiology score,reduced α wave and markedly increased δ wave on EEG,reduced amplitudes of evoked potentials P1,and significantly prolonged latency of S-P1 and NI-P1.In survived septic rats,6 had changes on neurological behaviors,EEG and somatosensory evoked potentials,and thus were diagnosed as SAE.The incidence of SAE was 46％.Conclusions SAE can be diagnosed in early stage by using neurobiology score,EEG and somatosensory evoked potentials,confirming the SAE rat models to be successfully established.