1. High mobility group box chromosomal protein 1 promotes IL-18 production and IL-18 receptor expression in macrophages
Academic Journal of Second Military Medical University 2010;30(12):1353-1357
Objective: To investigate whether high mobility group box chromosomal protein 1 (HMGB1) can activate macrophages to secrete IL-18. Methods: Mouse peritoneal macrophages were cultured with HMGB1 for indicated period, then the mRNA and protein expression of IL-18 and IL-18 receptors were examined by real-time PCR, ELISA, and flow cytometric analysis. Toll-like receptor (TLR) 2 and 4 agonists (1 μg/ml Pam3Cys or 100 ng/ml LPS) were used as parallel controls. Results: Two hours after culture with HMGB1(100 ng/ml), IL-18 mRNA expression was greatly increased. The expression of IL-18 protein was also greatly increased 24 h after culture with HMGB1, and the expression of IL-18R was slightly up-regulated. Pam3Cys (1 μg/ml) and LPS(100 ng/ml) could promote the expression of IL-18 at both mRNA and protein levels in the macrophages, and could up-regulate IL-18 receptors. Conclusion: HMGB1 can directly activate macrophages to secrete IL-18 and to express IL-18 receptor, suggesting that IL-18 may mediate the proinflammatory activity of HMGB1.
4.Non-invasive visualization of tumors in the mouse liver using a novel nanoparticle contrast enhanced micro-CT imaging procedure
Boyin QIN ; Xiaonan ZHANG ; Hua YANG ; Wenjiang ZHOU ; Xiaohui ZHOU
Acta Laboratorium Animalis Scientia Sinica 2014;(6):22-27
Objective To establish an in vivo imaging method of normal or tumorous liver in mice by using a new type nanoparticle contrast agent, ExiTron nano 12000, coupled with micro-CT imaging.Methods Six 6-8-week old male C57BL/6J mice were randomly divided into group A and group group B, by intravenous injection of 50μL and 100μL Ex-iTron nano 12000, respectively.In vivo Micro-CT scans were performed before contrast agent injection, 3 minutes, 24 hours, 7, 14, 28 and 56 days after injection.To determine which dose is suitable for long-term studies, gray scale value a-nalysis was performed on selected region of interest ( ROI) in the left lobe and right anterior lobe of the liver, and the chan-ges of liver tissue contrast was monitored after ExiTron nano 12000 injection.Three male HBV transgenic mice bearing liver tumors ( group C) were intravenously injected with the determined dose of ExiTron nano 12000 and were monitored by mi-cro-CT scans as above described.At 56 days after ExiTron nano 12000 injection, the mice were sacrificed and liver sam-ples were taken for histological analysis.Results Cross-sectional images taken at various time points and the average gray scale value ( AGSV) analysis in the mouse liver revealed that the AGSV peaked at 24 hours after injection of contrast rea-gent and good contrast still presented in the livers within 56 days of observation for both groups, though group B showed a significantly higher contrast than group A (P<0.01).Those data indicated that the dose of group B (100μL) was better to maintain ExiTron nano 12000 in the liver of mice for a long time.Contrast-enhanced by 100μL of ExiTron nano 12000, the liver tumor nodules in the mice of group C could be clearly delineated by Micro CT imaging during a 56 days observa-tion.Histological analysis revealed atypical hyperplasia, enlarged nuclei with hyperchromasia and cell necrosis in the tumors.Conclusions An in vivo imaging method was established to non-invasively visualize mouse liver using micro-CT combined with nanoparticle-based contrast agent and this technology may be applied to a live imaging of murine primary liv-er tumors.
5.Effects of dobutamine on acute lung injury in rabbits of septic shock
Caizhi SUN ; Haidong QIN ; Hua SHEN ; Yang SONG ; Zheng ZHANG
Chinese Journal of Emergency Medicine 2014;23(12):1338-1343
Objective To explore the effect of different doses of dobutamine on acute lung injury (ALI) in rabbits with septic shock and to clarify the possible mechanism.Methods The rabbits model of septic shock was made by cecal ligation and puncture combined with intravenous injection of endotoxin,70 male New Zealand white rabbits were randomly divided into five groups (14 rabbits in each groups):shamc operation group (group A),ALI group (group B),dobutamine low-dose group (group C),dobutamine medium-dose group (group D) and dobutamine high-dose group (group E),7 rabbits from each group were sacrificed 3 h and 6 h after septic shock.The level of cyclic adenosine monophosphate (cAMP) in lung tissue was detected by ELISA.The expression of aquaporin 5 (AQP5) protein was determined by western blotting.The wet to dry weight (W/D) ratio was measured.The pathological and ultrastructural changes of lung tissue were evaluated by optical microscopy and electron microscope,and lung injury score was assessed.The differences among the different groups were analyzed by one-way ANOVA (LSD test).Results The level of cAMP and expression of AQP5 protein in lung tissue at 3 h and 6 h were dramatically lower in group B than those in group A (3.53 ±0.43) pmol/mLvs.(21.18 ±0.62) pmol/mL; (0.44 ± 0.04) pmol/mLvs.(0.99±0.06)pmol/mL; (2.71±0.56)pmol/mLvs.(21.78±0.62)pmol/mL; (0.29 ±0.05) pmol/mLvs.(0.91 ±0.06) pmol/mL; all P <0.001,while the W/D ratio was obviously higher in group B than those in group A (all P <0.001).Compared with group B,the level of cAMP and AQP5 protein expression in lung tissue were significantly increased at 6 h in group C (8.48 ±0.61) pmol./ mLvs.(2.71±0.56) pmol/mL,P<0.01; (0.49 ±0.04) pmol/mLvs.(0.29 ±0.05) pmol/mL,P=0.001 and at3 hand6 hin groupDandE (10.86±0.66) pmol/mLvs.(3.53±0.43) pmol/mL; (0.60±0.05) pmol/mLvs.(0.44±0.04) pmol/mL; (13.80±0.49) pmol/mLvs.(2.71±0.56) pmol/mL; (0.64 ± 0.03) pmol/mLvs.(0.29 ± 0.05) pmol/mL; (15.57 ± 0.60) pmol/mL vs.(3.53±0.43) pmol/mL; (0.91 ±0.05) pmol/mLvs.(0.44 ±0.04) pmol/mL; (19.30±0.42) pmol/mL vs.(2.71 ±0.56) pmol/mL; (0.89 ±0.08) pmol/mL vs.(0.29 ±0.05) pmol/mL; all P < 0.01,while the W/D ratio in group E was decreased obviously (P =0.002; P =0.001).Compared with group C and D,the level of cAMP and the expression of AQP5 protein at 3 h and 6 h in group E increased significantly (all P <0.01.The pathological and ultrastructural changes of lung tissue were more intensive in group B than those in group A and the lung injury scores were obviously higher (P <0.01).The degree of lung pathological and ultrastructural lesion was ameliorated after administration of dobutanmine.Additionally,histological scores decreased significantly (P < 0.01).Conclusions Our study demonstrated that dobutamine could improve ALI induced by endotoxin,the mechanism of protective effect may involve in increasing the level of cAMP and up-regulating the AQP5 protein expression,and high-dose dobutamine had better effects.
6.Effect of Ultrashort Waves Combined with Medication on Children with Cerebral Dysfunction Syndrome Accompanied with Pneumonia
Hua YAN ; Jun-ying WANG ; Hua-lin DUAN ; Peijun YU ; Huijia ZHANG ; Rong QIN ; Weihong YANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(10):850-851
ObjectiveTo observe the effect of ultrashort waves combined with medication on children with cerebral dysfunction syndrome (CDS) accompanied with pneumonia.Methods76 children with CDS accompanied with pneumonia were randomly divided into treatment group (n=38, treated with anti-inflammatory and ultrashort waves one time per day for ten days) and control group (n=38, treated only with anti-inflammatory). The effective rate and the days of rales vanished of children in two groups after treatment were compared.ResultsIn the treatment group, 14 cases cured, 20 cases got significant result, 4 cases were effective, total effective rate was 100%; in the control group, 10 cases cured, 14 cases got significant result, 9 cases were effective, 5 cases were invalid, total effective rate was 86.8%. There was a significant difference between therapeutic effects of two groups ( P<0.05).ConclusionThe ultrashort waves therapy is an effective method for the children with CDS accompanied with pneumonia.
7.Effect of early goal directed therapy on tissue perfusion in patients with septic shock
Yuan-Hua LU ; Ling LIU ; Xiao-Hua QIU ; Qin YU ; Yi YANG ; Hai-Bo QIU
World Journal of Emergency Medicine 2013;4(2):117-122
BACKGROUND: This study aimed to observe the effect of early goal directed therapy (EGDT) on tissue perfusion, microcirculation and tissue oxygenation in patients with septic shock. METHODS: Patients with early septic shock (<24 hours) who had been admitted to the ICU of Zhongda Hospital Affiliated to Southeast University from September 2009 through May 2011 were enrolled (research time: 12 months), and they didn't meet the criteria of EGDT. Patients who had one of the following were excluded: stroke, brain injury, other types of shock, severe heart failure, acute myocardial infarction, age below 18 years, pregnancy, end-stage disease, cardiac arrest, extensive burns, oral bleeding, difficulty in opening the mouth, and the onset of septic shock beyond 24 hours. Patients treated with the standard protocol of EGDT were included. Transcutaneous pressure of oxygen and carbon dioxide (PtcO2, PtcCO2) were monitored and hemodynamic measurements were obtained. Side-stream dark field (SDF) imaging device was applied to obtain sublingual microcirculation. Hemodynamics, tissue oxygen, and sublingual microcirculation were compared before and after EGDT. If the variable meets the normal distribution, Student's t test was applied. Otherwise, Wilcoxon's rank-sum test was used. Correlation between variables was analyzed with Pearson's product-moment correlation coefficient method. RESULTS: Twenty patients were involved, but one patient wasn't analyzed because he didn't meet the EGDT criteria. PtcO2 and PtcCO2 were monitored in 19 patients, of whom sublingual microcirculation was obtained. After EGDT, PtcO2 increased from 62.7±24.0 mmHg to 78.0±30.9 mmHg (P<0.05) and tissue oxygenation index (PtcO2/FiO2) was 110.7±60.4 mmHg before EGDT and 141.6±78.2 mmHg after EGDT (P<0.05). The difference between PtcCO2 and PCO2 decreased significantly after EGDT (P<0.05). The density of perfused small vessels (PPV) and microcirculatory flow index of small vessels (MFI) tended to increase, but there were no significant differences between them (P>0.05). PtcO2, PtcO2/FiO2, and PtcCO2 were not linearly related to central venous saturation, lactate, oxygen delivery, and oxygen consumption (P>0.05). CONCLUSION: Peripheral perfusion was improved after EGDT in patients with septic shock, and it was not exactly reflected by the index of systemic perfusion.
8.The application of in vivo multi-channel recording methods in the studies of the affective pain in rats.
Zhen-hua LI ; Yang YANG ; Miao-miao HOU ; Yuan WANG ; Xia QIN ; Che ZHANG ; Yu ZHANG
Chinese Journal of Applied Physiology 2016;32(1):41-45
OBJECTIVETo record the electrical activities of Antirior cingulate cortex (ACC) neurons by in vivo multi-channel recording methods using the model of complete freund's adjuvant (CFA) induced conditioned place avoidance (C-CPA), which has been set up in our previous studies.
METHODSThe electrode was self-made and the CPA responses were recorded by in vivo multi-channel recording method.
RESULTS(1) The electrical activities of ACC neurons could be successfully recorded by the self-made electrode. (2) Before or after the injection of CFA, rats were respectively conditioned to the different place. The firing rates of ACC neurons in the CFA-paired place vs that in the non-CFA-paired place was (0.853 ± 1.377) imp/s vs (0.221 ± 0.971) imp/s (P < 0.05, n = 26). (3) The CPA responses in the CFA-paired place vs that in the non-CFA-paired place were (303.55 ± 61.77)s vs (140.32 ± 33.52)s(P < 0.05, n = 6).
CONCLUSIONThe firing rates of rACC (rostral Anterior Cingulate Cortex) neurons were involved in the occurrence of the affective pain.
Animals ; Electrodes ; Freund's Adjuvant ; Gyrus Cinguli ; cytology ; Neurons ; cytology ; Pain ; diagnosis ; Pain Measurement ; methods ; Rats ; Rats, Sprague-Dawley
9.Changes of glucose metabolism in mouse models of intracerebral hemorrhage:18 F-FDG microPET/CT study
Fanhui YANG ; Qin WANG ; Chunyin ZHANG ; Hua TAN ; Chaoxian YANG ; Wei WANG ; Qiang YOU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;(6):483-486
Objective To investigate the brain glucose metabolism with 18 F?FDG microPET/CT in mouse models of intracerebral hemorrhage (ICH). Methods A total of 12 healthy adult male mice were randomly divided into sham operation group (group A, n=6) and ICH model group (group B, n=6) by simple random sampling method. The animal models were established by injecting collagenase Ⅳ into the caudate nucleus of mice. Thereafter (5.5±0.3) MBq of 18F?FDG was injected into caudal vein at 6 h, 24 h, 48 h and 3 d, 5 d, 8 d, 14 d, respectively, following anesthesia. 18 F?FDG microPET/CT scans were ac?quired 30 min after the trace injection. SUV in the perihematomal brain tissue of ICH was measured and an?alyzed. Two?sample t test was used to compare SUV between groups. Results ( 1) Some mice had mild neurologic deficit after the sham operation in group A, while all mice had a marked neurologic deficit in group B, especially at 24 h after 18 F?FDG injection. ( 2) After 6 h, FDG uptake in perihematomal brain tis?sue decreased(SUV=0.80±0.04), which significantly lower than that in the opposite side(SUV=1.10± 0?04;t=2.69, P<0.05) and decreased to the minimum at 24 h(SUV=0.50±0.05). 18F?FDG uptake in perihematomal brain tissue began to increase at 3 d(SUV=1.20±0.05) and kept increasing during the 14 d observation. Compared with the group A, glucose metabolism in group B was significantly lower at each time point(t=37.67-86.60, all P<0.05). Conclusions 18 F?FDG microPET/CT may dynamically reflect the changes of brain glucose metabolism in ICH mouse models. The FDG uptake in the center of ICH may disap?pear and the volume of hematoma with decreased uptake may shrink during the observation period.
10.Effects of intestinal ischemia reperfusion on the progression of inflammatory reaction of hemorrhagic necrosis pancreatitis
Lin LIU ; Hua YU ; Qi QIN ; Jungang ZHANG ; Yang LIU ; Shichang DENG ; Gang ZHAO ; Chunyou WANG
Chinese Journal of Digestive Surgery 2011;10(5):362-365
Objective To investigate the effects of intestinal ischemia reperfusion (IIR) on the progression of inflammatory reaction in hemorrhagic necrosis pancreatitis (HNP).Methods Eighty rats were randomly divided into sham operation (SO) group,acute edematous pancreatitis (AEP) group,AEP + IIR group and HNP group according to the random number table.Erythrocyte velocity (EV),functional capillary density (FCD) and leukocyte adherence (LA) were observed at 0,1,2,3 and 6 hours after the models were completed.The serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected.All data were analyzed by using the analysis of variance or the t test.Results The level of EV in the AEP group significantly decreased at 1 hour,and got increased at 3 hours,while the level of EV in the AEP group was still significantly lower than that in the SO group ( t =9.60,P < 0.05 ).The levels of EV in the AEP + IIR group and HNP group constantly decreased,and increased at 6 hours,but were continually lower than that in the AEP group ( t =6.03,6.12,P <0.05 ).The level of FCD in the AEP group was significantly lower than that in the SO group at 3 hours ( t =8.20,P<0.05).The levels of FCD in the AEP + IIR group and HNP group were significantly lower than that in the AEP group at 3 hours (t =35.60,23.80,P < 0.05 ).Compared with AEP group,the level of LA in the AEP group was significantly increased at 1 hour ( t =75.00,P < 0.05 ) and reached peak at 3 hours.The levels of LA in the AEP + IIR group and HNP group were significantly higher than that in the AEP group at 1,2,3,6 hours (t =23.00,29.50,53.00,38.70,23.10,48.20,39.20,47.50,P<0.05).Compared with SO group,the level of TNF-α in the AEP group significantly increased since l hour (t =77.00,P < 0.05),and began to decrease at 3 hours; the levels of TNF-α in the AEP +IIR group and HNP group at 2 hours were significantly higher than that in the AEP group (t =23.50,18.10,P<0.05).The levels of IL-6 in the AEP group at 1,2,3,6 hours were significantly higher than those in the SO group ( t =93.50,146.00,243.60,209.20,P < 0.05 ).The levels of IL-6 in the AEP + IIR group and HNP group at 1 hour were not significantly different from that in the AEP group ( t =2.30,2.03,P > 0.05),while the levels of IL-6 in the AEP + IIR group and HNP group at 2 hours were significantly higher than that in the AEP group (t =35.63,29.80,P < 0.05 ).Conclusion IIR may enhance the inflammatory reaction of AEP and IIR might be one of the factors to exaggerate the inflammatory reaction of HNP.