Objective To investigate the changes of immune state and the impact on immunological rejection elicited by abdominal cavity bacterial infection after DA-Lewis rat liver transplantation.Methods Orthotopic liver transplant model was established by modified Kamada two-cuff technique.The animals were divided randomly into Group 1,isotonic Na chloride injected into abdominal cavity 3 days after operation;Group 2,mixed Bacillus coli liquid injected instead of saline;Group 3,immunosuppressive drug CsA administered routinely after operation(3 mg?kg-1?d-1).All the animals were sacrificed 7 days after infection.The blood and graft samples were collected for cell-subpopulation,mixed lymphocyte culture,IL-4,IFN-? mRNA detection and histological evaluation.Results Seven days after infection,the lympholeukocyte population,CD4/CD8(G1=1.753?0.181,G2=1.384?0.073,G3=0.997?0.025)and lympholeukocyte function(SI:G1=67.59?3.40,G2=37.14?0.90,G3=15.87?0.51)declined in Group 2 as compared with other groups and cellular differentiation drifted to Th2.There was significant difference between Group 2 and Group 1 or 3.Conclusion Abdominal cavity bacterial infection after rat liver transplantation will promote the differentiation of T cells into Th2,down-regulate CD4/CD8 ratio and immune function of lymphocytes and accordingly alleviate partly the acute rejection following liver transplantation.

Objective To establish a rat model of peritoneal bacterial infection after liver transplantation.Methods To construct a dark Agouti rat-to-Lewis(DA-to-LEW) rat model of liver transplantation.Peritoneal bacterial infection in the rats was induced by intraperitoneal injection of bacterial suspension.The liver function,blood gas,blood cell count and other indicators of the rat models were detected.Results There was a high mortality rate in rats with bacterial injection at day 5 after liver transplantation,therefore unfavorable for the following study.It waft better to inject the bacteria in an amount of 5×10~5 cfu/mL at day 3 after liver transplantation.The cumulative 7-day survival rate of those rats after infection reached up to 37.5%.The infection became increasingly severe,the general conditions were worsening,the rectal temperature was rising,the WBC count was increased,the pH was decreased,liver dysfunction was progressively increased,and metabolic acidosis occurred in the rats.Liver parenchymal damage was more pronounced than that of bile ductal injuries,and the rats died one after another at about 5 days after infection.Pathological examination of multiple organs showed that the main cause of death of the rats was liver damage,without accompanying lung and kidney damages.Conclusion The results of this study suggest that it is a successful method to establish a rat model of peritoneal bacterial infection after liver transplantation,and this model can be used in related experimental researches.

Objective To evaluate the effects of modified early goal directed therapy (EGDT )on the prognosis of patients with septic shock .Methods Clinical data of 116 patients with septic shock admitted to ICU during January 2011 to March 2013 were retrospectively analyzed .Patients were divided into modified early goal‐directed therapy group (n=57) and traditional early goal‐di‐rected therapy group (n=59) according to different methods of treatment ,the patients′28‐day survival rates of these 2 groups were compared .Modified early goal‐directed therapy are divided into survival group (n=46) and non‐survival group (n=11) according to 28‐day prognosis .Acute physiology and chronic health evaluation Ⅱ (APACHEⅡ ) score ,sequential organ failure assessment (SOFA) ,multiple organ dysfunction syndrome (MODS) score and other relevant indicators of survival group and non‐survival group were compared .Results The 28‐day survival rate in modified early goal‐directed therapy group had increased approximately 18 .9% higher than that of the traditional early goal‐directed therapy group(P< 0 .05) .The APACH Ⅱ score ,SOFA score and MODS score in non‐survivors were significantly higher than those of survivors in modified EGDT group ,which were[(29 .36 ± 1 .57)d vs .(24 .30 ± 3 .27)d] ,[(13 .45 ± 0 .52)d vs .(12 .78 ± 1 .33)d] ,[(9 .00 ± 0 .00)d vs .(4 .04 ± 1 .94)d]separately .And vaso‐pressors time and mechanical ventilation time was significantly longer in non‐survivors than survivors(P<0 .05) .Conclusion Mod‐ified early goal directed therapy could improve 28‐day survival rate ,and it show s beneficial effects on outcome of critical patients w ith septic shock .

Objective To analyze the epidemiology and clinical characteristics of epidemic encephalitis B in Yuneheng area of Shanxi Province in 2006.Methods Seventy-nine cases diagnosed with epidemic encephalitis B were enrolled in this study and correlated data,epidemic and clinical characteristics,laboratory examinations and treatment outcome were analyzed.Results Sixty-two of 79 patients(78.5%)were middle-aged or old people,all cases(100%)occurred in July,August and September,69 cases(87.3%)were peasants.All patients(100%)had fever,73(92.4%)had conscious disturbance,27(34.2%)had respiratory failure.Encephalitis B specific IgM antibody was examined and 40 cases(85.1%)were positive.Twenty cases(25.3%)had complications.When they were discharged,37 cases(46.8%)recovered completely,14 cases(17.7%)died,12 were improved and 16 were voluntarily discharged,7 cases(8.9%)left more or less neurological deficits. Conclusion The epidemiology and clinical characteristics are important basis to diagnose epidemic encephalitis B.

AIM : To investigate the effects of the new K ATP channel opener, iptakalim (Ipt), on the Parkinsonian symptoms induced by haloperidol in rats. METHODS : Using L -DOPA as the positive drug, the influences of Ipt on latency time, walking time, rearing and grooming of the parkinsonian-like rats induced by haloperidol were observed. RESULTS : Ipt ameliorated hypolocomotion and assuaged catalepsy induced by haloperidol in rats. Moreover, the effects of Ipt ( 0.5 mg?kg -1) were better than those of L -DOPA (100 mg?kg -1). CONCLUSION : Opening of K ATP channels may play a key role in improving Parkinsonian symptoms induced by haloperidol in rats.

Objective: To observe the changes in numbers and degranulation status of local mast cells (MCs) of the complementary acupionts, as well as the concentrations of serum histamine and bradykinin after acupuncture at rat's complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 10), for exploring the mechanisms of the synergistic effect of complementary acupionts. Methods: Using random number table method, 40 Wistar rats were randomly divided into a blank control group (group K), an acupuncture at Yanglingquan (GB 34) group (group A), an acupuncture at Yinlingquan (SP 9) group (group B), an acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group AB), and an acupuncture at points near Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group CD), 8 rats in each group. Group A received acupuncture at Yanglingquan (GB 34); group B received acupuncture at Yinlingquan (SP 9); group AB received acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9); group CD received acupuncture at the control points [points 3 mm away from Yanglingquan (GB 34) and Yinlingquan (SP 9) respectively]. Acupuncture was performed at bilateral points in each group. Rats in group K were fixed using the same method as rats in the other 4 groups without acupuncture stimulation. Needles of 0.35 mm in diameter and 40 mm in length were used for acupuncture. Needle handle was connected to the G6805-Ⅱ mode electroacupuncture (EA) device after needling qi was obtained, with sparse-dense wave, frequency of 2 Hz/100 Hz and current of 1 mA to keep the needle handles slightly tremulous while the rats kept quiet. Rats were continuously stimulated by EA for 20 min each time. Experimental interventions were conducted on the 1st, 3rd, 5th and 7th days when the experiment started, for 4 times in total. Specimens from rats in each group were collected 2 h after the 7th day. The levels of histamine and bradykinin in serum were detected by enzyme-linked immunosorbent assay (ELISA). Local tissues of the points were used to prepare cryosections. The changes of MCs were observed after toluidine blue staining. Results: Compared with group K, the numbers and degranulation rates of MCs in group A, group B, group AB and group CD were significantly increased (all P<0.05). The number and degranulation rate of MCs in group AB were significantly higher than those in group A, group B and group CD (all P<0.05). The order of histamine levels from high to low was: group AB >group B > group A > group CD > group K, and the differences were statistically significant (all P<0.05); the levels of bradykinin in group AB, group A and group B were significantly higher than those in group K and group CD (all P<0.05). There was no significant difference in the bradykinin level between group A and group B, nor between group CD and group K (all P>0.05). Conclusion: The number and degranulation rate of MCs of the complementary acupionts are significantly increased after acupuncture at complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 9) of rats, and the serum levels of histamine and bradykinin are increased, which may be one of the mechanisms of the synergism produced by the combination of complementary acupionts.

Mechanical ventilation (MV) with large tidal volumes can increase lung alveolar permeability and initiate inflammatory responses,resulting in ventilator-induced lung injury (VILI).The mechanisms of the injurious effects of MV and the genetic susceptibility remain unclear.VILI-related genes such as cysteine-rich angiogenic inducer 61 (Cyr61)have been demonstrated to play a detrimental role in the aggressive ventilation strategies.In the present study,we investigated the involvement of Cyr61 in the VIM and the underlying mechanism.A549 cells were exposed to cyclic stretch of varying durations and then the mRNA and protein levels of Cyr61 were measured by real-time PCR and Western blotting,respectively.Additionally,after exposure ofA549 cells to cyclic stretch for 5 min to 1 h,the expression levels of nuclear factor kappaB (NF-κB) and IL-8 were detected by ELISA and Western blotting.Thereafter,Cyr61 expression was depressed in A549 cells with the siRNA pGenesill.1-Cyr61-3 before the cyclic stretch,and IL-8 secretion and the activation of NF-κB pathways were probed by ELISA and Western blotting,respectively.Moreover,a NF-κB inhibitor (PDTC) and an activator (TNF) were used before mechanical stretch.Realtime PCR and ELISA were performed to detect the mRNA and protein of IL-8,respectively.The results showed that the mechanical cyclic stretch led to increased Cyr61 expression at mRNA and protein levels in A549 cells.Additionally,cyclic stretch also mobilized NF-κB from the cytoplasm to the nucleus and increased IL-8 secretion in A549 cells.The inhibition of Cyr61 blocked the NF-κB activation and IL-8 secretion in response to cyclic stretch.Inhibition of NF-κB attenuated the mRNA and protein expression of IL-8 in A549 cells transfected with Cyr61 siRNA.It was suggested that Cyr61/NF-κB signaling pathway mediates the upregulation of IL-8 in response to cyclic stretch in A594 cells.These findings support the hypothesis that Cyr61 plays a critical role in acute lung inflammation triggered by mechanical strain.

BACKGROUND:Human umbilical vein endothelial cells transfected with lentivirus containing enhanced green fluorescent protein can be easily traced. The optimal multiplicity of infection and time for producing strong fluorescence intensity can lay the foundation of tracing human umbilical vein endothelial cells in animal models.
OBJECTIVE:To observe expression of lentivirus containing enhanced green fluorescent protein in human umbilical vein endothelial cells, and thereby to find a stable method to label human umbilical vein endothelial cells.
METHODS:Using 0.1%col agenase perfusion digestion, we isolated human umbilical vein endothelial cells, which then were placed into a culture medium containing 20%fetal bovine serum and endothelial cellgrowth factor and observed under an inverted microscope. Fol owing digestion, centrifugation and suspension, the cells were counted and divided into four groups, 5.0×105 cells in each group. After cells were seededonto 24-wel plates, 10μL serum-free Dulbecco’s modified Eagle’s medium was added into the blank group, and lentiviruses containing enhanced green fluorescent protein were added into another three groups for celltransfection respectively at multiplicities of infection of 2, 3, 4. There were three dishes in each group.
RESULTS AND CONCLUSION:After cultured for 5-7 days, isolated cells grew into a single layer and exhibited a cobblestone-like arrangement under a light microscope. In addition, factor VIII related antigen test was positive. A green fluorescence was visible at 24 hours of transfection, and peaked at 72 hours. Transfection efficiency was in a linear growth with the multiplicity of infection. Up to the 21st day of transfection, the green fluorescence was stil visible. After 0, 7, 14, 21 days of transfection, the number of human umbilical vein endothelial cells showed no difference between the transfection group with the multiplicity of infection=3 and blank group, suggesting the proliferative ability of cells has no changes after transfection with lentivirus containing enhanced green fluorescent protein. These findings indicate that the lentivirus containing enhanced green fluorescent protein can highly transfect human umbilical vein endothelial cells, and green fluorescent protein can sustainably express for 21 days but cannot impact the cellproliferation.

Objective To evaluate the efficacy and safety of linezolid in the treatment of severe lung infections caused by methicillin-resistant Staphylococcus aureus (MRSA).Methods Fifteen patients admitted to ICU due to severe lung infection caused by MRSA received linezolid treatment. WBC, lactic acid (LAC), IL-1β, IL-6, and TNF-α levels were measured before and after treatment. Results Clinical efficacy rate was 73.3%. The level of WBC, LAC and inflammatory cytokines decreased significantly after linezolid treatment (P<0.01).Conclusions Linezolid shows good efficacy in the treatment of severe lung infections caused by MRSA.

Objective To test plasma levels of uPA,uPAR,D-dimer,IL-6 and TNF-α,and observe the relationships between uPA,uPAR and D-dimer,IL-6,TNF-αin patients with SIRS.Methods A prospective,clinical case-control study was adopted.Cases were collected from our hospital in January 2008 to January 2010,and all were ＞ 55 years of age.Venous blood samples were collected via routine venipuncture.Eighty-five patients were divided into two groups according to diagnostic criteria of SIRS:SIRS group collected from Intensive Care Unit ( n =50) and non SIRS group collected from medical ward ( n =35).The control group comprised 30 unrelated healthy blood donors who visited the General Health Checkup Division at our hospital.Patients with (1) pregnant women; (2) cancer; (3) died after admitted into ICU in 7 days; (4) after cardiopulmonary resuscitation ; ( 5 ) with previous blood system diseases; (6) patients with SIRS before admitted into ICU were excluded from the study.uPA,uPAR,D-D,IL-6 and TNF-α in blood were detected by commercial enzyme-linked immunosorbent assay (ELISA) kits.The data was analyzed using SPSS version17.0.Data accorded with normal distribution of measurement was expressed as mean ± standard,and analyzed by independent-samples t test; non-normal distribution of measurement data,expressed by median,was analyzed with Mann-Whitney test.Relationships between plasma levels of uPA,uPAR and D-dimer,IL-6 TNF-α were analyzed using Spearman rank correlation test.To compare with blood level of uPA,uPAR,IL-6 and TNF-α in SIRS patients in the application of diagnostic value of MODS,we constructed the receiver operating characteristic curve ( ROC curve) for blood levels of uPA,uPAR,IL-6 and TNF-α in 24 h.Results The plasma levels of uPA,uPAR,D-dimer,IL-6 and TNF-αin patients of SIRS were obviously higher compared with non-SIRS and normal controls ( all P ＜ 0.01 ).Correlation analysis showed that there was positive correlation between uPAR level and IL-6 level (r =0.395,P =0.004) ; there was positive correlation between uPAR level and TNF-αlevel ( r =0.606,P ＜0.01 ).There was no correlation between uPAR levd and D-dimer level ( r =- 0.069,P =0.632).There was no correlation between uPA level and D-dimer,IL-6 or TNF-α ( all p ＞ 0.05).There ROC curve were made based on the abilities of uPAR,D-dimer,IL-6 and TNF-αlevels in 24 hours to diagnose MODS,and the ROC areas under the curves were 0.76,0.58,0.86,0.83 respectively.Conclusions These results demonstrate that uPA and uPAR play a major contributory role in patients with SIRS in the process of coagulation disorders,but the mechanism in SIRS is not the same.uPAR may play a central rolein the development of SIRS to MODS.