1.Serum levels of TNF-?, IL-6, IL-10, CRP and D-dimer in patients with multiple organ dysfunction syndrome
Zhijie HE ; Chun CHEN ; Longyuan JIANG ; Yini SHI
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To determine the changes of the serum levels of tumor necrosis factor ? (TNF-?), interleukin-6 (IL-6), IL-10, C reactive protein (CRP) and D-dimer in the patients with multiple organ dysfunction syndrome (MODS) and compare the relationship between the levels of cytokines in early stage and MODS. METHODS: The serum values of TNF-?, IL-6, IL-10, CRP and D-dimer were measured in 27 patients with MODS in 1 d, 3 d and 5 d after undergoing disease, and compared with the adult peripheral blood of 15 normal controls. The levels in the first undergoing day between the lived group (n=19) and died group (n=8) were compared. RESULTS: The serum levels of TNF-?, IL-6, IL-10, CRP and D-dimer in MODS group were higher than that in control (P
2.Effects of ulinastatin on expression of intestinal defensin-5 mRNA in the rat model of sepsis
Longyuan JIANG ; Meng ZHANG ; Jianxing CHANG ; Zhengfei YANG ; Zhengchao LUO
Chinese Journal of Emergency Medicine 2008;17(8):848-851
Objective To investigate the effeets of ulimstatin on expression of intestinal defemin-5 mRNAin the rat model of sepsis.Method The experiment was performed in pharmaco-laboratory of medical college,Sun Yat-Sen University.sixty Sprague-Dawley rals were randomly divided into control,sepsis,pretreated andtreated groups(n=15).Semis was induced in the mts of latter three groups by cecal lifo.and puncture(CLP).The rats of pretreated group received 25 000 U/kg ulinastatin 2 hours before operation and the rats of uli-nastatin treated groups received 50 000 U/kg ulinastatin 2 hours after operation.Some pieces of ileum mucosa weretaken 12 h after CLP.Tge pathological changes were observed and the expression of RD-5 mRNA was detectedwith RT-PCR.All data were managed by SPSS 13.0 software and arIaIyzed by using One-way ANOVA and LSD-ttest.Results The expression of RD-5 mRNA in the rats of sepsis group significantly decreased compared to col-trol(P<0.05).The expression of RD-5 mRNA of pretreated and treated groups sigificantly inereased comparedto sepsis group(P<0.05);pretreated groups had more increased expression of,RD-5 mRNA compared to treatedgroups(P<0.05).Conclusions The expression of intestinal RD-5 mRNA significantly decreases in sepsis,which could be improved by the treatment of ulinadtatin leading to intestinal mucosal protection of the siqnifleant.The pretreatment may be more effective than the theTapeatic treatment in the rat model of sepsis.
3.Analysis of emergency medical rescue system of general hospital during spring festival with snow disaster
Weiyi QIN ; Longyuan JIANG ; Lidian HUA ; Shaohui TANG ; Xuefei TAO
Chinese Journal of Emergency Medicine 2008;17(6):571-574
Objective To explore the strategy of emergency medical rescue of the massive crowd in general hospital during spring festival with snow disaster. Method The clinical data of 20 966 emergency cases were analyzed retrospectively from 22 Jan,2008 to 6 Feb, 2008 with snow disaster, and concerned about the ratio of different diseases, the character of pre-hospital care and the contrast between emergency medical treatment and routine work. Results The accidence of respiratory disease ( 57.3 % ) was followed by gastrointestinal ( 25.5 % ) and trauma (6.2% )during the emergency medical treatment, and surgical trauma, syncope, coma and convulsion were the most common symptoms, also in some conditions, but empties returning was 30.3% . Similar to the above situation, the extremities (56%)and head injury (24%)were most commonly in the hospital emergency department. The incidence of falling accidents was high( 35.7 % ), and two of them were dead due to trauma on died of being trampled, and on the other was electrothermal burn and falling. Conclusions The general hospital is very important in emergency medical treatment, and it should be ready to tackle the emergency disaster, in order to reduce the loss to minimum.
4.Minocycline inhibits BV-2 cell activation by regulating P2X7 receptor
Shuqiong LIU ; Lianhong YANG ; Longyuan JIANG ; Jinhao YE
Chinese Journal of Pathophysiology 2014;(6):1066-1069
AIM: To explore the role of P2X7 receptor in inhibition of lipopolysaccharide (LPS)-stimulated BV-2 cell activation by minocycline .METHODS:BV-2 cells were divided into 5 groups:control group, LPS group, LPS+0.1 μmol/L Mino group, LPS+1 μmol/L Mino group and LPS+10 μmol/L Mino group.The expression of P2X7 re-ceptor was determined by real-time PCR and Western blotting .The levels of TNF-αand IL-1βin the microglia culture su-pernatants were measured by ELISA .The morphological changes of the cells were also observed .RESULTS: After ex-posed to LPS, the expression of P2X7 receptor increased in BV-2 cells at mRNA and protein levels .The concentrations of TNF-αand IL-1βin the microglia culture supernatants also increased .Meanwhile, 0.1~10μmol/L minocycline inhibited those changes in a dose-dependent manner .CONCLUSION:Minocycline inhibits the activation of microglia .The mecha-nism may be related to the P2X7 receptor.
5.Effects of Xuebijing injection on intestinal mucosal morphology in rats with sepsis
Lili ZHOU ; Ruiming CHANG ; Longyuan JIANG ; Zitong HUANG
Chinese Journal of Emergency Medicine 2009;18(7):707-710
Objective To observe the effect of injecting Xuebijing via different routes, as either treatment or pretreatment, on changes in intestinal mucosal morphology in a rat model of sepsis. Method Ninety-one healthy Sprague-Dawley rats were randomly divided into five groups: 1) control group ( n = 7) , 2) sepsis group ( n = 21) , 3) intragastric pretreatment group ( n = 21) , 4) intravenous pretreatment group (n = 21) , and 5) intravenous treatment group (n = 21) . Except for the control group, the other groups were further divided into three sub-groups for assessment at 3, 6 and 12 h post-operation ( n = 7 per group) . Sepsis was induced by cecal ligation and puncture (CLP) . For the intragastric pretreatment group, Xuebijing injection (5 mL/kg) was administered via intragastric injection 2 hours before CLP. For the intravenous pretreatment group, Xuebijing injection (5 mL/kg)was administered via the caudal vein 2 hours before CLP. For the intravenous treatment group, Xuebijing injection (5 mL/kg) was intravenously infused 2 hours after CLP. The control group received no treatment. The ileum was removed from all rats for measurement. The rats were sacrificed at 3, 6 or 12 h after operation to obtain the ileum.Intestinal mucosal damage index and morphological changes in the intestinal mucosa were detected by light microscopy and electron microscopy. Analysis of variance was used to compare the groups. P -values < 0.05 were considered to indicate statistically significant differences. Results Intestinal mucosal damage was significantly reduced in and the three treatment groups compared with the untreated sepsis group (3h, F =53.35; 6h, F =74.93; 12 h, F - 171.27; P =0.000). Intestinal damage was significantly reduced in the intravenous pretreatment group compared with the intragastric pretreatment group (3 h, F = 53.35,P =0.036; 6 h, F = 74.93,P =0.039; 12 h, F = 171.27, P =0.042). Conclusions Irrespective of the route, the administration of Xuebijing protected against intestinal mucosal damage and intravenous pretreatment exerted the most effective protection against intestinal mucosal damage in this rat model of sepsis.
6.Protective effect of ulinastatin on cerebral tissue in septic rats
Chaotao ZENG ; Meng ZHANG ; Tianen ZHOU ; Longyuan JIANG
Chinese Journal of Emergency Medicine 2014;23(8):885-889
Objective To explore the effect of Ulinastatin on blood brain barrier (BBB) and apoptosis of neural cells in septic rats.Methods Fifty-two clean level male Sprague-Dawley rats were randomly (random number table) divided into six groups:Sham groups at 6 h and 24 h,each group with six rats.Sepsis groups (CLP) and Ulinastatin treated groups (UTI) at 6 h and 24 h,each group with ten rats.In CLP and UTI groups,cecal ligation and puncture (CLP) were performed to induce sepsis.Sham group was only opened and closed abdomen.Ulinastatin (50 000 U/kg) was administered via femoral vein 1 h after CLP.The same volume of saline instead of Ulinastatin was administered in Sham and CLP groups.The neurological status was assessed by Neurological Deficit Scale Scores (NDSS) at 6 h and 24 h after CLP.Then the brain was harvested for HE staining and weighing water content.The BBB permeability was assayed by Evans Blue dye extravasations.Apoptosis of neural cells were detected by TUNEL immune fluorescence.Statistical analysis was performed with SPSS version 13.0,ANOVA was used for multiple groups comparison and t-test for paired comparison.Results The Neurological Deficit Scale Scores of UTI group was lower than Sham group (P < 0.05) but higher than that of CLP group (P < 0.05).Swelling,degeneration and edema were observed in cerebral cortex and hippocampal neurons in CLP group through light microscope,and were more serious than those in UTI group.Compared with UTI 24 h group,BBB permeability of CLP 24 h group significantly rose (P < 0.05).The number of apoptosis of neural cells increased more in CLP group than it did in UTI group (P < 0.05).Conclusions Ulinastatin could protect the cerebral tissue in septic rats by alleviating the damage of BBB and reducing the apoptosis of neural cells.
7.The effect of thrombolytic therapy and TOAST and OCSP classification on the short-term prognosis of patients with acute ischemic stroke
Jie ZHANG ; Yunyuan HUANG ; Zhengfei YANG ; Longyuan JIANG
Chinese Journal of Emergency Medicine 2021;30(2):204-207
Objective:To investigate the short-term prognosis of patients with acute ischemic stroke after thrombolysis with alteplase and the relationship between TOAST and OCSP classification and prognosis of patients with acute ischemic stroke.Methods:A retrospective analysis was conducted in 193 patients with acute ischemic stroke in the Emergency Department of our hospital from January 2019 to June 2020, including 158 males and 85 females, aged 69.46±13.02 years. Among them, 66 patients were treated with alteplase thrombolysis and 127 patients were not treated with thrombolysis. The short-term prognosis of thrombolytic patients and non-thrombolytic patients were compared; the influence of TOAST classification and OCSP classification on the prognosis of acute ischemic stroke was analyzed.Results:There was no significant difference in the length of hospital stay between the thrombolytic group and the non-thrombolytic group ( P>0.05), while the improvement rate of NIHSS score, GCS score, and mRS score were significantly different at admission and discharge ( P<0.05). According to TOAST classification, 78 cases (40.41%) were LAA, 100 cases (51.81%) were SAO, 9 cases (4.66%) were CE, 3 cases (1.55%) were SOE, and 3 cases (1.55%) were SUE; SAO type accounted for the highest proportion (51.81%), the improvement rate of NIHSS score at admission and discharge was significantly different from that of the LAA patients ( P<0.05), and the short-term prognosis was the best. According to OCSP classification, 39 cases (20.21%) were LACI, 64 cases (33.16%) were PACI, 55 cases (28.50%) were POCI, 35 cases (18.13%) were TACI, among which the PACI patients had the highest proportion, while the improvement rate of NIHSS score in the LACI patients was significantly different at admission and discharge compared with other types of patietns ( P<0.05), and the short-term prognosis was good. Conclusions:The short-term prognosis and symptom improvement of patients with acute ischemic stroke after alteplase thrombolytic therapy are obvious. TOAST classification and OCSP classification have certain prediction effects and play clinical reference roles on the short-term prognosis of patients with acute ischemic stroke.
8.Effect of autophagy regulator on the injury of rat hippocampal neurons induced by oxygen-glucose deprivation
Tianen ZHOU ; Chaotao ZENG ; Jiajun FANG ; Longyuan JIANG ; Tao YU
Chinese Critical Care Medicine 2017;29(8):738-743
Objective To explore the effect of autophagy regulator on the injury of rat hippocampal neurons induced by oxygen-glucose deprivation (OGD).Methods Rat hippocampal neurons were cultivated in primary and subjected to OGD to simulate neuronal hypoxic ischemia injury for 2 hours or 6 hours followed by reperfusion for 12 hours with or without 3-methyladenine (3-MA, 20μmol/L) or rapamycin (0.2μmol/L). The morphology of neurons was observed with optical microscope. The expression of autophagy-related protein (LC3, P62) and apoptosis-related protein (cleaved caspase-3) were assessed by Western Blot analysis. The apoptosis of neurons was detected by flow cytometry, the release rate of lactate dehydrogenase (LDH) was calculated by automatic biochemical analyzer, and the cell activity was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay.Results Compared with the control group, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.091±0.160, 3.422±0.186 vs. 0.256±0.021), cleaved caspase-3 (gray value: 0.230±0.025, 0.440±0.051 vs. 0.050±0.007), neuronal apoptotic rate, LDH release rate [(38.50±4.15)%, (59.60±5.65)% vs. (12.40±1.32)%] were increased, while the expression of P62 (gray value: 0.290±0.025,0.120±0.026 vs. 0.450±0.040), neuronal activity [(71.40±7.23)%, (42.80±4.12)% vs. (100.30±2.30)%] were decreased at 2 hours or 6 hours after OGD (allP < 0.05). When the time of OGD was 2 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ (gray value: 2.281±0.121), the neuronal activity [(51.10±5.73)%] were decreased, while the expression of P62 and cleaved caspase-3 (gray scale: 0.410±0.037, 0.330±0.027, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (47.30±4.43)%] were increased (allP < 0.05). When the time of OGD was 2 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.689±0.214), the neuronal activity [(85.30±8.56)%] were increased, while the expression of P62 and cleaved caspase-3 (gray value: 0.170±0.040, 0.090±0.096, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (24.30±2.14)%] were decreased (allP < 0.05). On the contrary, when the time of OGD was 6 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.021±0.178, 0.240±0.017), neuronal apoptotic rate, the injury of neurons [LDH release rate: (36.60±3.45)%] were decreased, while the expression of P62 (gray value: 0.350±0.060), the neuronal activity [(59.70±6.13)%] were increased (allP < 0.05). When the time of OGD was 6 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.923±0.201, 0.590±0.062), neuronal apoptotic rate, the injury of neurons [LDH release rate:(71.20±7.81)%] were increased, while the expression of P62 (gray value: 0.070±0.008), the neuronal activity [(27.30±2.12)%] were decreased (allP < 0.05).Conclusion The enhancement of autophagy has protective effect on neurons under the condition of mild OGD, while it can aggravate the injury of neurons induced by a long-time OGD.
9.Protective effects of ulinastatin on intestinal barrier damaged after cardiopulmonary resuscitation in rats
Ruiming CHANG ; Jianxing CHANG ; Zhipeng JIANG ; Liqiang WEN ; Kai YU ; Tao YANG ; Longyuan JIANG
Chinese Journal of Emergency Medicine 2015;24(11):1234-1238
Objective To investigate the protective effects of Ulinastatin on intestinal barrier damaged after cardiopulmonary resuscitation (CPR) in rats in order to illustrate the possible mechanism.Methods Twenty-one male SD rats were divided into three groups randomly (random number) including control group (sham group, n =7), cardiopulmonary resuscitation group (CPR group, n =7) and ulinastatin group (UTI group, n =7).The rats were anesthetized with pentobarbital sodium (45-60 mg/kg) by intraperitoneal injection.The rats of sham group were only treated with endotracheal intubation.Ulinastatin (100 000 U/kg) were injected via caudal vein 2 hours prior to CPR, and cardiac arrest was made in rats and cardiopulmonary resuscitation was carried out in the UTI group, while equivalent volume of sterile saline was used instead in the CPR group.Blood and ileum samples were obtained at 48 hour after restoration of spontaneous circulation (ROSC).The levels of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) were assayed by ELISA (enzyme-linked immunosorbent assay), the protein levels of caspase-3 were determined by western blot, the intestinal mucosa were stained by terminaldeoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and ileac mucosa were observed under transmission electron microscope.Data were processed with SPSS 17.0 software.Results The plasma levels of TNF-α and IL-1β were dramatically higher in CPR group than those in other two groups (CPR vs.sham, P < 0.01;CPR vs.UTI, P < 0.05).Moreover, the tight junctions between cells obviously broadened and loosened in the CPR group were found under electron microscope, however, this phenomenon was not obvious in the UTI group.A large number of apoptotic cells were observed by TUNEL assay in the CPR group, but a small number of apoptotic cells were observed in the UTI group.The protein levels of caspase-3 in the UTI group were higher than those in sham group, but lower than those in CPR group (both P < 0.05).Conclusions Ulinastatin has protective effects on the intestinal barrier damaged after cardiopulmonary resuscitation in rats by decreasing the proinflammatory mediators in the blood, reducing the expression of caspase-3and then reducing the numbers of apoptotic intestinal cells.
10.The experimental probe into the construction of oxygen glucose deprivation/reoxygenation model of hippocampal neurons of rat in vitro
Jun JIANG ; Yue FU ; Xiangshao FANG ; Jianxing CHANG ; Longyuan JIANG ; Zitong HUANG
Chinese Journal of Emergency Medicine 2010;19(5):497-501
Objective To establish the oxygen glucose deprivation (OGD)/reoxygenation experimental model of hippocatnpal neurons of rat in vitro, and to try to identify the length of time for producing optimum injury in this model. Method The primary hippocampal neurons of newborn Sprague-Dawley rats were cultured for 7 days and randomly (random number) divided into a control group and OGD groups. The OGD groups were assigned into 1 h, 2 h, 4 h, 6h, 8 h and 10 h subgroups in accordance with different lengths of time for oxygen glucose deprivation. The neurons of OGD groups were placed into a tri-gas incubator containing 0.5% oxygen and the culture medium was substituted with the glucose-free Earle' s balanced salt solution, simulating cerebral ischemia injury in vivo. The morphology of neurons was observed after reoxygenation for 24 hours. The MIT assay was used to determine the rate of survived cells derived from the value of optical density (OD) of cells. The lactate dehydro-genase (LDH) content in culture medium was detected to evaluate the neuron injury. The apoptotic rate of neurons was measured by using flow cytometry. Dunnett-test and Spearman correlation analysis were used to analyze the data with SPSS version 16.0 soft ware package. Results The morphological damage of neurons in OGD groups aggravated gradually, optical density and cell survival rate decreased (rs= -0.961 and rs = -0.966, P <0.01), and the amount of LDH increased (rs = 0.990, P <0.01) with longer duration of exposure to oxygen glucose deprivation, and the rate of neuron apoptosis increased obviously which was significantly statistical difference in com-parison with the control group (P < 0.05). Under the setting of oxygen glucose deprivation for 6 hours, the apop-tosis rate of neurons approximated to 50% . Conclusions The oxygen glucose deprivation/reoxygenation model of rat's hippocampal neurons in vitro was established successfully. From the findings of morphological changes and apoptosis rate of neurons, the oxygen glucose deprivation for 6 hours may be the suitable length of time for inducing neuron injury in this model.