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Sepsis is a life-threatening organ dysfunction caused by dysregulation of the host response due to infection, and it can further develop into septic shock.Currently, sepsis is still a leading cause of death in children all over the world.Therefore, early assessment of the severity and prognosis of sepsis is of great significance.However, there are no indexes with high sensitivity and specificity for evaluating the severity and prognosis of sepsis at present.In recent years, a large number of studies have revealed the essential role of platelets in sepsis.It has been reported that the platelet count is an independent factor affecting the severity and prognosis of sepsis patients.Up to now, the specific mechanism of sepsis-induced thrombocytopenia has not been fully clarified.In this review, the value of thrombocytopenia in predicting the severity and prognosis of sepsis patients was elaborated.
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Objective To invkstigatk thk kffkcts of intkrlkucin-6(IF-6)on mitochondrial biogknksis in ac-tivatkd astrocetks(LS)and thk rolk of adknosink monophosphatk protkin cinask( LMPK)in this prockss. Methods Thk LS isolatkd from nkonatal rat ckrkbral codkx wkrk purifikd and culturkd. Thk LS was randomle dividkd into 5 groups:control group,lipopolesaccharidk(FPS)+intkrfkron-γ(IPN-γ)group( IPN-γ group),FPS+IPN-γ+IF-6 group(IF-6 group),FPS+IPN-γ+IF-6A siANL+IF-6 group(siANL group),and FPS+IPN-γ+nkga-tivk control(NC)+IF-6 group(NC group),thkn,LS in kach group was trkatkd for 6 h. Tumor nkcrosis factor-α (TNP-α)mANL and intkrlkucin-1β(IF-1β)mANL kxprkssion wkrk dktkctkd be adopting rkvkrsk transcription-polemkrask chain rkaction(AT-PCA). Thk lkvkls of rkactivk oxegkn spkciks(AOS)wkrk dktkctkd be fluorksknt probk mkthod and thk lkvkls of adknosink triphosphatk(LTP)wkrk dktkctkd be lucifkrask mkthod. Ckll viabilite was kvaluatkd be using ckll count Kit-8. Pkroxisomk prolifkrator-activatkd rkckptor gamma coactivator-1α(PGC-1α),nuclkar rk-spiratore factor-1(NAP-1),mitochondrial transcription factor L( TPLM)and phospho-adknosink monophosphatk activatkd protkin cinask(p-LMPK)protkin kxprkssion wkrk dktkctkd be using Zkstkrn blotting. ResuIts (1)Com-parkd with thk control group,thk mANL kxprkssions of TNP-α and IF-1β(2. 548 ± 0. 154 vs. 1. 000 ± 0. 001,P﹦ 0. 000;2. 912 ± 0. 102 vs. 1. 000 ± 0. 001,P﹦0. 000),thk lkvkls of AOS[(245. 307 ± 13. 379)APR vs.(69. 460 ± 7. 257)APR,P﹦0. 000]and LTP[(1. 558 ± 0. 008)nmol╱mg protkin vs.(1. 016 ± 0. 025)nmol╱mg protkin,P﹦0. 000]significantle klkvatkd,and thk ckll viabilite(0. 840 ± 0. 013 vs. 1. 000 ± 0. 001,P﹦0. 000)dkcrkaskd,whilk thk protkin kxprkssion of NAP-1(0. 406 ± 0. 045 vs. 0. 157 ± 0. 016,P﹦0. 017),TPLM(0. 605 ± 0. 025 vs. 0. 416 ± 0. 013,P﹦0. 005)klkvatkd in FPS+IPN-γ group,and thk diffkrkncks wkrk significant(all P＜0. 05).(2)Comparkd with FPS+IPN-γ group,thk lkvkls of LTP[(1. 763 ± 0. 028)nmol╱mg protkin vs.(1. 558 ± 0. 008)nmol╱mg pro-tkin,P﹦0. 000],thk ckll viabilite(0. 910 ± 0. 024 vs. 0. 840 ± 0. 013,P﹦0. 008)wkrk klkvatkd,whilk thk protkin kx-prkssion of PGC-1α(0. 724 ± 0. 027 vs. 0. 586 ± 0. 039,P﹦0. 000),NAP-1(1. 036 ± 0. 211 vs. 0. 406 ± 0. 045,P﹦0. 000),TPLM(0. 786 ± 0. 058 vs. 0. 605 ± 0. 025,P﹦0. 002)and p-LMPK(1. 094 ± 0. 223 vs. 0. 755 ± 0. 084,P﹦0. 014)wkrk klkvatkd in IF-6 group,and thk diffkrkncks wkrk significant( all P＜0. 05).(3)Comparkd with IF-6 group,LTP[(1. 187 ± 0. 005)nmol╱mg protkin vs.(1. 763 ± 0. 028)nmol╱mg protkin,P﹦0. 000]and thk ckll viabili-te(0. 680 ± 0. 040 vs. 0. 910 ± 0. 024,P ﹦0. 000)all dkcrkaskd in siANL group,whilk thk protkin kxprkssion of PGC-1α(0. 631 ± 0. 022 vs. 0. 724 ± 0. 027,P﹦0. 020),NAP-1(0. 386 ± 0. 066 vs. 1. 036 ± 0. 211,P﹦0. 000), TPLM(0. 593 ± 0. 022 vs. 0. 786 ± 0. 058,P﹦0. 009)and p-LMPK(0. 365 ± 0. 063 vs. 1. 094 ± 0. 223,P﹦0. 002) significantle dkcrkaskd in siANL group,and thk diffkrkncks wkrk significant(all P＜0. 05). ConcIusions IF-6 can incrkask mitochondrial biogknksis in activatkd LS,which is probable mkdiatkd through up-rkgulating thk kxprkssion of LMPK.
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Objective To preliminarily investigate the role of Vitamin C in cecal ligation and puncture-induced septic brain injury.Methods Male specific pathogen free (SPF) Sprague-Dawley male rats were randomly assigned into control group,sham operation group,sepsis group and sepsis therapy group.The rats in sepsis group were prepared by cecal ligation and puncture.The rats in sepsis therapy group were injected sodium ascorbate through the tail vein 3 h after the cecal ligation and punature procedure.The animals in other groups were subjected only to subcutaneous bolus injection of 9 g/L saline only.Animals were evaluated by neurologic reflex scores before sacrifice and brain tissues were quickly removed at the indicated time points.Reactive oxygen species (ROS),superoxide dismutase (SOD),malondialdehyde (MDA),nitric oxide (NO),inducible nitric oxide synthase (iNOS) and catalase (CAT) were determined by using enzyme assay kits.Hematoxylin-eosin (HE) staining was used to observe morphological changes in brain tissues.Results The survival rate of the sepsis group (30％ at 7th day) was significantly lower than that of the control group (100％ at 7th day)and sham operation group(100％ at 7th day).The survival rate of the sepsis therapy group (45％ at 7th day)was significantly higher than that of the sepsis group(P ＜ O.05).The neurological reflex assessment began to decrease at 6 h in sepsis group and reached the lowest at 24 h (6.00 ± 0.53).The sepsis therapy group (7.62 ± 0.52) was significantly higher (P ＜ 0.05) than the sepsis group and began to recover at 72 h (8.63 ±0.52).ROS,SOD,MDA,NO and iNOS in the sepsis group and the sepsis therapy group reached a peak at 24 h,which decreased at 72 h.The value in sepsis therapy group was significantly decreased than that in the sepsis group,and the difference was statistically significant(P ＜0.05).CAT changed in the opposition.The SOD/CAT in sepsis group was the highest 24 h after the operation,while the ratio in sepsis therapy group was significantly improved.SOD/CAT and MDA were positively correlated(r =0.968,P ＜ 0.05).HE staining showed significant damage to the brain tissue structure in the sepsis group,however some improvement was observed in the sepsis therapy group.Conclusion Vitamin C can significantly improve the survival rate and encephalopathy prognosis in the cecal ligation and puncture SD rat models.The mechanism may be related to the reduction of oxidative stress.
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The definition of sepsis has been updated as life-threatening organ dysfunction caused by a dysregulated host response to infection according to the Third International Consensus Definitions for Sepsis and Septic Shock (SEPSIS-3).Sepsis affects functions of endothelial cell (EC) which include vasoregulation,barrier function,inflammation,hemostasis,and is thought to be the key factor in the progression from sepsis to organ failure.Recent studies also demonstrated the mechanism of endothelial dysfunction in sepsis is mediated by glycocalyx shedding.This review covers the current insight in sepsis-associated endothelial dysfunction in different organ systems,as well as the clinical assessmeut and clinical trial aiming at the system of endothelium.
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The definition of sepsis has been updated as life-threatening organ dysfunction caused by a dysregulated host response to infection according to the Third International Consensus Definitions for Sepsis and Septic Shock (SEPSIS-3).Sepsis affects functions of endothelial cell (EC) which include vasoregulation,barrier function,inflammation,hemostasis,and is thought to be the key factor in the progression from sepsis to organ failure.Recent studies also demonstrated the mechanism of endothelial dysfunction in sepsis is mediated by glycocalyx shedding.This review covers the current insight in sepsis-associated endothelial dysfunction in different organ systems,as well as the clinical assessmeut and clinical trial aiming at the system of endothelium.
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Objective To investigate the protective effects and mechanism of insulin(INS) on brain in septic rats,and explore the possible role of uncoupling protein 2 (UCP2) in these effects.Methods Fifty male specific pathogen free(SPF) Sprague-Dawley rats were randomly divided into normal control (CN) group(n=10),lipopolysaccharide(LPS) group(n=20) and INS group (n=20) according to random number table.The septic rat model was established through an intraperitoneal injection of 15 mg/kg LPS of gram-negative bacteria.The rats in the INS group received a 1 U/kg INS injection subcutaneously 30 minutes before the injection of LPS,and the rats in the CN group were given equivalent 9 g/L saline in the same way.Eight rats in each group were killed,and their cerebral cortex were collected after the injection of LPS for 24 h.Pathological change of cerebral cortex was detected by Hematoxylin-Eosin(HE) staining.The cerebral cortex mitochondia were extracted for detecting the levels of reactive oxygen species(ROS),malondialdehyde (MDA) and the activity of superoxide dismutase(SOD).Neuronal apoptosis was detected by terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling staining.UCP2 mRNA expression was detected by quantitative real-time(RT)-PCR.Apoptosis-associated protein B lymphocyte tumor-2(Bcl-2),Bcl-2 associated X protein(Bax),cleaved cysteinyl aspartate specific protease(cleaved Caspase-9) and UCP2 protein expression were determined by Western blot.Results (1)Compared with the CN group,obvious abnormal pathological change was revealed by HE staining in cerebral cortex of rats in the LPS group and the INS group,but the pathological change was attenuated in the INS group compared with the LPS group.(2)Compared with the CN group,the levels of mitochondrial ROS[(210.01±14.09) RFU vs.(49.06±7.28) RFU] and MDA[(2.19±0.18) nmol/mg pro vs.(1.25±0.11)nmol/mg pro]in the LPS group significantly increased,whereas SOD activity significantly decreased [(238.49±35.60) U/g pro vs.(446.66±24.90)U/g pro],and the differences were significant(all P<0.05).Compared with the LPS group,the levels of ROS [(152.69±15.83) RFU vs.(210.01±14.09) RFU] and MDA[(1.55±0.14) nmol/mg pro vs.(2.19±0.18) nmol/mg pro] in the INS group decreased,while SOD activity increased[(327.8±23.26) U/g pro vs.(238.49± 35.60) U/g pro],and the differences were significant(all P<0.05).(3)Compared with the CN group,the neuronal apoptosis index of cortex in the LPS group was elevated[(54.16±6.84)% vs.(5.45±1.43)%],while the expression of Bcl-2 decreased (627±0.018 vs.0.739±0.020),but the expressions of Bax(0.768±0.019 vs.0.520±0.010) and cleaved Caspase-9(0.739±0.016 vs.0.467±0.030) increased,and the differences were significant(all P<0.05).Compared with the LPS group,the neuronal apoptosis index of cortex in the INS group decreased [(33.30±3.07)% vs.(54.16±6.84)%],but the Bcl-2 expression increased (0.743±0.022 vs.0.627±0.018),and Bax (0.687±0.034 vs.0.768±0.019) and cleaved Caspase-9(0.551±0.013 vs.0.739±0.016) were reduced,and the differences were significant (all P<0.05).(4)Compared with the CN group,the mRNA (2.248±0.155 vs.1.000±0.100) and protein expression of UCP2 (0.659±0.016 vs.0.599±0.018) were elevated in the LPS group.Compared with the LPS group,the UCP2 mRNA (2.944±0.117 vs.2.248±0.155) and UCP2 protein (0.719±0.018 vs.0.659±0.016) increased,and the differences were significant(all P<0.05).Conclusions INS can protect the brain of septic rats through alleviating mitochondrial oxidative stress and inhibiting the mitochondrial-initiated apoptotic pathway to reduce neuronal apoptosis.INS upregulates UCP2 expression in the brain of septic rats,which may play a role in the protective effects mentioned above.
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Objective Mitochondrial dysfunction, cell energy metabolism, and oxidative stress play important roles in sepsis-induced acute brain injury.This study was to investigate the effects of Xingnaojing Injection (XNJ) on brain mitochondrial oxidative stress in rats with lipopolysaccharide (LPS)-induced sepsis.Methods Totally, 252 male SD rats were randomly divided into a normal control group, 3 LPS-induced sepsis model groups (LPS 6, 24, and 48 h), and 3 XNJ treatment groups (XNJ 6, 24, and 48 h), with 36 in each group.After treatment, the mitochondrial membrane potential (MMP) was monitored by flow cytometry and the levels of manganese superoxide dismutase (Mn-SOD), malondialdehyde (MDA), nitric oxide (NO), and nitric oxide synthase (NOS) were determined by chromatometry.Results The MMP was significantly increased in the XNJ 6 h group as compared with the LPS 6 h group (0.80±0.11 vs 0.54±0.19, P<0.05).In the LPS and XNJ groups, the levels of MDA and NOS reached the peak at 6 hours and then dropped gradually, while those of NO and Mn-SOD rose to the peak at 24 hours followed by a gradual fall.Statistically significant differences were observed in the levels of MDA, NOS and NO between the LPS 6h and XNJ 6 h groups (P<0.05), as well as in those of NOS, NO and Mn-SOD between the LPS 24 h and XNJ 24 h groups (P<0.05).Conclusion Xingnaojing Injection can elevate the level of the brain mitochondrial membrane potential, improve anti-oxidation indexes in the mitochondria, and protect brain mitochondria in sepsis rats.
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Objective To research the protective effect of insulin(IN)on lipopolysaccharide(LPS)- induced impairments of rat cardiomyocytes H9c2,and the role of uncoupling protein 2(UCP2)in this process. Methods Using randomized controlled grouping,after cultured for 24 h,H9c2 cells were randomly divided into 5 groups as follows:con-trol group,LPS stimulation group(LPS group),LPS + 70 IU/ L IN group(IN 70 IU/ L group),LPS + 350 IU/ L IN group(IN 350 IU/ L group),and LPS + 700 IU/ L IN group(IN 700 IU/ L group). H9c2 cells in IN group were treated with 70 IU/ L,350 IU/ L or 700 IU/ L IN 15 min before LPS stimulation,and H9c2 cells in control group were treated with an equal volume of saline. After that,cells in group LPS and IN were treated with LPS for 24 h. Lactate dehydro-genase(LDH)in the culture was determined with LDH detecting assay kit. The activity of reactive oxygen species (ROS)and superoxide dismutase(SOD),and content of malonaldehyde(MDA)were determined by colorimetric detec-tion. Cell viability was evaluated by cell count kit - 8. The expressions of UCP2 in transcription and translation levels were detected through transcription polymerase chain reaction and Western blot respectively. Results The levels of LDH,MDA,and intracellular ROS in LPS group significantly increased compared with control group[LDH:(829. 3 ± 75. 3)U/ L vs(223. 5 ± 23. 6)U/ L,MDA:(60. 90 ± 5. 73)nmol/ mgprot vs(19. 70 ± 1. 99)nmol/ mgprot,ROS:(410. 2 ± 81. 6)U/ well vs(94. 3 ± 18. 5)U/ well,all P ﹤ 0. 05)],while the cell viability and SOD activity significantly decreased[cell viability:0. 822 ± 0. 058 vs 1. 012 ± 0. 023,SOD:(49. 20 ± 5. 81)U/ mgprot vs(89. 80 ± 2. 57)U/ mg-prot,all P ﹤ 0. 05]. And the mRNA and protein expressions of UCP2 in LPS stimulation group were up - regulated (1. 867 ± 0. 130 vs 1. 028 ± 0. 097,0. 288 ± 0. 018 vs 0. 180 ± 0. 008,all P ﹤ 0. 05). 350 IU/ L and 700 IU/ L IN inter-vention significantly decreased the levels of LDH,MDA and intracellular ROS[LDH:(568. 2 ± 35. 7)U/ L,(622. 8 ± 27. 6)U/ L vs(829. 3 ± 75. 3)U/ L,MDA:(29. 20 ± 4. 20)nmol/ mgprot,(42. 10 ± 2. 32)nmol/ mgprot vs(60. 90 ± 5. 73)nmol/ mgprot,ROS:(270. 3 ± 46. 8)U/ well,(301. 5 ± 16. 9)U/ well vs(410. 2 ± 81. 6)U/ well,all P ﹤ 0. 05], increased the cell survival and the levels of SOD activity[cell viability:0. 960 ± 0. 029,0. 906 ± 0. 039 vs 0. 822 ± 0. 058,SOD:(75. 20 ± 2. 21)U/ mgprot,(61. 20 ± 3. 38)U/ mgprot vs(49. 20 ± 5. 81)U/ mgprot,all P ﹤ 0. 05]. And IN with 350 IU/ L and 700 IU/ L increased the mRNA and protein expression of UCP2(3. 830 ± 0. 265,2. 855 ± 0. 215 vs 1. 867 ± 0. 130,0. 464 ± 0. 215,0. 355 ± 0. 006 vs 0. 288 ± 0. 018,all P ﹤ 0. 05). Compared with 70 IU/ L and 700 IU/ L IN group,350 IU/ L IN group had better results. Conclusions IN attenuates LPS - induced oxidative injury in H9c2 cells,which is probably mediated through up - regulating the expression of UCP2.
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Objective To investigate the differential expression of microRNA - 30e in sepsis - induced acute lung injury(ALI)and its correlation with interleukin(IL)- 1β and tumor necrosis factor(TNF)- α from two aspects of in vivo and in vitro. Methods Thirty SD male rats were randomly divided into 5 groups:normal control group,3 - hour sepsis group,6 - hour sepsis group,12 - hour sepsis group and 24 - hour sepsis group in equal number. Sepsis - in-duced ALI model was induced by intraperitoneal injection of lipopolysaccharide(LPS,10 mg/ kg). The rat alveolar mac-rophages NR8383 were divided into blank control group and LPS(1 mg/ L)stimulated 3,6,12,24 hour groups. Inverse transcription - polymerase chain reaction was used to assay the production changes of IL - 1β,TNF - α and miRNA - 30e in lungs and cells. The injury of lung tissue was evaluated through histopathology. Results The levels of IL - 1β and TNF - α in lung tissues of rats in sepsis groups were obviously up - regulated when compared with those in normal control groups(all P ﹤ 0. 01). The lung tissue hematoxylin - eosin staining indicated ALI in the sepsis group. The relative expression of miR - 30e in rat lung tissue in sepsis 3,6,12,24 hour groups were respectively 0. 26 ± 0. 02, 0. 41 ± 0. 08,0. 29 ± 0. 05 and 0. 18 ± 0. 05,which were significantly lower than those in normal control group(1. 23 ± 0. 24,all P ﹤ 0. 01). The levels of IL - 1β and TNF - α in LPS stimulated NR8383 cells at different time points were obviously up - regulated when compared with those in blank control groups(all P ﹤ 0. 01). The relative expression of miR - 30e in LPS stimulated 3,6,12,24 hour groups were respectively 0. 27 ± 0. 04,0. 55 ± 0. 05,0. 65 ± 0. 02 and 0. 41 ± 0. 10,which were significantly lower than those in blank control group(1. 17 ± 0. 21,all P ﹤ 0. 01). The expres-sion of miR - 30e in lung tissues of groups showed significantly negative correlations with those of IL - 1β and TNF - α(IL - 1β:r = - 0. 417,P = 0. 022;TNF - α:r = - 0. 437,P = 0. 016). The expression of miR - 30e in LPS stimulated NR8383 cells of groups also showed significantly negative correlations with those of IL - 1β and TNF - α(IL - 1β :r =- 0. 713,P = 0. 003;TNF - α:r = - 0. 712,P = 0. 002). Conclusions The expression level of miR - 30e was signifi-cantly down - regulated in sepsis - induced ALI,and had a significantly negative correlation with IL - 1β and TNF - α, which may be used as a new biomarker of diagnostic,prognosis evaluation and therapy of sepsis - induced ALI.
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Uncoupling protein 2(UCP2)is a proton transporter which presents in the mitochondrial inner membrane. Recent studies have found that UCP2 plays important roles in protecting mitochondria functions,re-ducing mitochondrial ROS generation by inducing proton leak across the inner mitochondrial membrane and pro-moting mild uncoupling which leading to a decrease in the mitochondrial inner membrane′s potential. Because of its antioxidant function,UCP2 has been studied in several domains. This review provides novel insights into the molecular mechanisms,by which the activity of UCP2 is regulated and describe novel findings concerning basical experiments of UCP2.
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Objective To investigate the effects of epinephrine in sepsis-associated lung injury in rats. Methods Thirty SD rats were randomly divided into three groups(n =10 per group):control group received intravenous 0. 9% saline 2. 4 ml/( kg·h ); LPS group received intravenous LPS ( 6 mg/kg ); epi-nephrine treatment group received an infusion of epinephrine 0. 6μg/( kg·min) after LPS intravenous injec-tion . Blood samples were taken at 2 h and 6 h after LPS injection and the levels of serum tumor necrosis factor ( TNF)-α,interleukin( IL)-6 and IL-10 were detected. The rats were sacrificed at 6 h. The lung tissues and bronchoalveolar lavage fluid( BALF) were collected. Pathological changes of the lung tissues were observed under light microscope. Water content of lung,expression of TNF-α,IL-6 and IL-10 in BALF and in serum were detected. Results (1) The water content of lung in LPS group significantly increased compared with that in control group(85. 24% ± 5. 87% vs. 70. 19% ± 5. 87%) and epinephrine group(78. 00% ± 6. 41%) (P<0. 05). (2)Pathological examination showed that LPS could cause pulmonary capillary hyperemia,ede-ma,inflammatory cells infiltration. Atelectasis and alveolar edema were found in small number of lung tissue. Compared with LPS group, epinephrine ameliorated the lung pathological injury. ( 3 ) Compared with LPS group,serum levels of TNF-α and IL-6 significantly decreased ( P <0. 05 ) , whereas IL-10 increased ( P <0. 05) in epinephrine group. (4)Compared with LPS group,BALF levels of TNF-α[(78 ± 9)ng/L vs. (102 ±16)ng/L]andIL-6[(268±42)ng/Lvs.(347±50)ng/L]significantlydepressed(P<0.05),whereas BALFlevelsofIL-10[(210±23)ng/Lvs.(146±34) ng/L]elevated(P <0.05) inepinephrinegroup. Conclusion Epinephrine could reduce the acute lung injury caused by LPS. Its protective effect may be re-lated to decreasing the levels of TNF-α and IL-6,elevating IL-10 level.
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Sepsis associated encephalopathy (SAE)is the most common form of encephalopathy in the pediatric intensive care units and might appear before other systemic features of sepsis.The pathogenesis of SAE is complex and not clear.SAE causes increased morbidity and mortality but has limited therapeutic options.SAE has become a hot issue in critical care medicine.
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ObjectiveTo study the change of the pulmonary surfactant protein C, D (SP-C, SP-D) and apoptosis of alveolar epithelium cells in neonatal rats with hyperoxia-induced lung injury.MethodsThe neonatal rats born within 24 hours were divided into the air group (n=50) and the hyperoxia group (n=50). The lung tissue was collected on the ifrst, third, seventh, tenth, fourteenth day after the hyperoxia exposure. The pathological changes were observed by HE staining. The apoptosis rate of lung epithelial cells was detected by TUNEL (terminal deoxyn ucleotidyl transfer-mediated end labeling). The content of SP-C and SP-D in broncho alveolar lavage lfuid (BALF) was detected by enzyme-linked immunosorbent assay.ResultsIn the air group, as age increased, the alveolar were gradually more completely formed with the regular shape and uniform size. Mean-while, in the hyperoxia group, as age increased, the number of alveolar was reduced, the small blood vessels expanded, the alve-olar hemorrhage was increased, the interstitial cells were increased and the lung tissue was swelling. The levels of SP-C, SP-D decreased with the increase of age in the air group. The level of SP-C in hyperoxia group was lower than that in the air group on the ifrst day. It was higher than that in the air group on the third day, peaked on the seventh day, and then it began to decline on the tenth day and decreased more obviously on the fourteenth day. The level of SP-D in hyperoxia group was not signiifcantly dif-ferent from that in the air group on the ifrst day, was higher than that in the air group on the third day and peaked on the seventh day. Then it began to decline on the tenth day and decreased more on the fourteenth day. ConclusionsLong-term inhalation of high concentrations of oxygen inhibits alveolar development. With the prolonged time of oxygen inhalation, the apoptosis of lung epithelial cells is increased, and the level of SP-C and SP-D in BALF was increased ifrst and then decreased.
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Objective To study the reasonable doses, efficacy and safety of regional citrate anticoagulation (RCA) for continuous veno-venous hemofiltration(CVVH) in children. Methods There were 66 patients hospi-ta-lized in Pediatric Intensive Care Unit of Zhujiang Hospital,Southern Medical University treated with RCA-CVVH that were recruited in the study from October 2012 to July 2014. The patients were divided into 4 groups according to their weight:≤10 kg( group Ⅰ) ,20 kg≥weight>10 kg( group Ⅱ) ,30 kg≥weight>20 kg( group Ⅲ) ,>30 kg( groupⅣ),and each group randomly received 2 different doses of anticoagulant acid citrate dextrose formula A(ACD-A):ACD-A(mL/h)=0. 75×blood flow rate(BFR)(mL/min)(A dose) and ACD-A=1. 5×BFR(B dose). Data of hemo-filter duration, activated partial thromboplastin time( APTT) ( systemic and circuit) , ionized calcium( Ca2+) ( systemic and circuit), blood urea nitrogen(BUN), serum creatinine(Cr), alanine aminotransferase(ALT), aspartate amin-otransferase(AST), blood pH, sodium ion(Na+), bicarbonate ion(HCO3-) were collected and analyzed. Results There was no significant difference in BUN,Cr,ALT,AST and APTT of 2 different doses of ACD-A among the groups (all P>0.05);pH of B dose of ACD-A in group Ⅰwas significantly higher than that in A dose(F=7.384,P=0. 015);pH of B dose of ACD-A in groupⅡwas significantly higher than that in A dose(F=4. 492,P=0. 046),HCO3-of B dose of ACD-A in groupⅠwas significantly higher than that in A dose(F=7. 735,P=0. 013);HCO3-of B dose of ACD-A in groupⅡwas significantly higher than that in A dose(F=4. 644,P=0. 042);hemofilter duration of B dose of ACD-A in group Ⅲ was significantly higher than that in A dose(t=-3. 147,P=0. 016);hemofilter duration of B dose of ACD-A in groupⅣwas significantly higher than that in A dose(t=-6. 342,P=0. 000). Conclusions RCA-CVVH is effective and safe for critical children,and different doses of ACD-A for children with different weight can re-duce metabolic alkalosis and enhance regional anticoagulation.
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Objective To investigate the protective effect of continuous intravenous infusion of Isoproterenol (ISO)on myocardial mitochondria of early septic rats and the corresponding mechanism.Methods Thirty Sprague Dawley (SD)rats were randomly divided into 5 groups (6 cases per group):control group,endotoxin group,ISO small-dose group,ISO medium-dose group and ISO large-dose group.Endotoxin group and ISO intervene group received same management apart from drug intervention:receiving intravenous injection of lipopolysaccharide (LPS)10 mg/kg followed by an continuous intravenous infusion of 9 g/L saline 1 mL/h or ISO 0.06 μg/(kg · min),0.30 μg/(kg · min)and 0.60 μg/(kg · min).Control group received intraperitoneal injection and continuous intravenous infusion with the same amount of 9 g/L saline.The primary endpoint of the study was 24 hours after injection of 9 g/L saline or LPS.Serum creatine kinase (CK) and creatine kinase isoenzyme (CK-MB),oxidative and nitrosative stress levels and swelling of isolated heart mitochondrion were detected.The pathological changes of the myocardium and morphologic changes of the heart mitochondria were observed through light microscope and scanning electron microscope,respectively.Results The levels of CK,CK-MB,nitric oxide (NO) content,inducible nitric oxide synthase (iNOS) activity and malondialdehyde (MDA)content in endotoxin group were increased compared with control group (all P ＜ 0.05),while the superoxide dismutase (SOD) activity decreased [(11.543 ± 1.080) U/mg prot vs (9.892 ±0.815) U/mg prot,P ＜0.05].The morphology of the heart mitochondria significantly changed (such as swelling,disordered arrangement,crest fracture,fusion and cavitations,and so on).ISO intervention significantly decreased the levels of CK,CK-MB and mitochondrial swelling (all P ＜ 0.05) and increased the SOD activity (all P ＜ 0.05).The levels of NO content,iNOS activity and MDA content were significantly decreased in small-dose group [(10.823 ± 2.240) μmol/g prot vs (7.917 ± 2.203) μmol/g prot,(0.045 ± 0.008) U/mg prot vs (0.033 ± 0.003) U/mg prot,(1.663 ± 0.618) mmol/mg prot vs (0.768 ± 0.312) mmol/mg prot,all P ＜ 0.05],while the levels of iNOS activity and MDA content were significantly increased in medium-and large-dose group (all P ＜ 0.05) ; compared with medium-dose group,the degree of mitochondrial swelling in large-dose group increased (1.160 ± 0.186 vs 1.393 ± 0.128,P ＜ 0.05).The pathological changes of the myocardium mitochondria significantly improved.Conclusions The myocardium and myocardial mitochondria of early septic rats were damaged,continuous intravenous infusion of low-dose ISO revealed protective effect on these damages,and the corresponding mechanism may relate to the decrease of the oxidative and nitrosative stress.
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ObjectiveTo investigate the effects of autophagy on exocrine function of pancreas in rats with acute sepsis, and to determine whether the mitochondrial coenzyme Q (Mito Q) can prevent exocrine dysfunction of pancreas mediated by autophagy.Methods ExperimentⅠ: 30 Sprague-Dawley (SD) rats were randomly divided into three groups, with 10 rats in each group. All the rats were given lipopolysaccharide (LPS, 10 mg/kg) intraperitoneally, and Wortmannin (2 mg/kg), the specific inhibitor of autophagy (LPS+ Wortmannin group), Mito Q (6.5μmol/kg, LPS+Mito Q group), or the same volume of normal saline (LPS group) was respectively injected via the tail vein 1 hour later. Survival rate was assessed within 12 hours after LPS injection. ExperimentⅡ: another 100 male SD rats were randomly divided into ten groups with 10 rats in each group: namely control 4, 6 and 12 hours groups, LPS 4, 6 and 12 hours groups, and LPS+ Wortmannin 4 hours group, Wortmannin 4 hours group, LPS+ Mito Q 6 hours group, and Mito Q 6 hours group. The protocols of model reproduction and drug administration were the same as in the experimentⅠ. Blood samples were collected at each time point, and the amylase content was determined with the velocity method. The levels of reactive oxygen species (ROS) in the pancreases were measured with enzyme-linked immunosorbent assay (ELISA). The expression of the autophagy-related protein LC3 was determined with Western Blot. The pathological changes in the pancreas were observed with microscopy.Results① The survival time in the LPS+ Wortmannin group was significantly shorter than that in the LPS group (hours: 7.50±0.64 vs. 11.90±0.13,χ2= 19.847,P= 0.001). There was no significant difference in the survival time between LPS+ Mito Q and LPS groups (hours: 11.60±0.24 vs. 11.90±0.13,χ2= 1.055,P= 0.137).② The serum amylase in the LPS 6 hours, LPS+ Wortmannin 4 hours, and LPS+ Mito Q 6 hours groups were significantly higher than those in the control group at the same time points (U/L:2 881.00±550.12 vs. 2 099.20±249.57, 3 672.00±779.24 vs. 2 081.36±245.18, 2 975.20±687.03 vs. 2 099.20± 249.57, allP 0.05). Light microscopy showed that obvious pathological changes were found in the pancreas in the LPS 6 hours and 12 hours groups, LPS+Wortmannin 4 hours group, and LPS+ Mito Q 6 hours group. Electron microscopy showed that the number of autophagic vacuoles increased 6 hours after LPS administration. There was no difference at any time point in the number of autophagic vacuoles between LPS+ Mito Q 6 hours group and LPS 6 hours group, and the autophagic vacuoles were not found after Wortmannin intervention. It was demonstrated by Western Blot that the levels of LC3 protein in the LPS 6 hours and 12 hours groups, and LPS+ Mito Q 6 hours group were significantly higher than those of the control group at the same time points (A value: 0.34±0.02 vs. 0.17±0.02, 0.37±0.03 vs. 0.18±0.04, 0.36±0.02 vs. 0.17±0.02, allP 0.05).Conclusions Autophagy prevents exocrine dysfunction of pancreas in septic rats, and the autophagic capacity or autophagosome-formation rate may determine the development of exocrine pancreatic dysfunction. The mitochondria-targeted antioxidant Mito Q does not prevent exocrine dysfunction of pancreas.
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Multiple organ dysfunction syndrome(MODS),a hot topic worldwide,has made rapid progress with nigh mortality.MODS in pediatrics versus MODS in adults are similar but different.Due to special age-related physiological characteristics.It is difficult to carry out randomized controlled clinical study compared with adults.Diagnosis and treatment of pediatric MODS can only be obtained with reference to adult MODS.This study reviews on the epidemiology,clinical scoring system,pathogenesis,clinical features and treatment of MODS in pediatrics.
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Objective To observe the effect of Xingnaojing injection in myocardial mitochondrial oxidative stress injury in sepsis mice. Methods The septic model were set up by receiving endotoxin through interaperitoneal injection. After Xingnaojing injection by gastric tube , seventy mice were randomly divided into 7 groups in ten of each group including group LPS-6 h , group LPS-24 h , group LPS-48 h; group XNJ-6 h , group XNJ-24 h, group XNJ-48 h and control group. The myocardial mitochondrial changes , the semi-quantitative scores and the level of SOD、MDA、NO、iNOS in sepsis mice were observed. Results Xingnaojing injection could improve the myocardial mitochondrial changes , reduce the semi-quantitative scores , significantly reduce the level of MDA、NO、iNOS and elevate the level of SOD. Conclusion Xingnaojing injection could significantly reduce the myocardial mitochondrial oxidative stress level and improve the ability of clearing oxygen radicals , thereby protect the myocardial mitochondrion in sepsis mice.
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Objective To investigate the effect of concurrent exercise intervention in metabolic endotoxemia induced by high-fat diet in rats,and further understand the damage of liver mitochondrial ultramicrostructure.Methods Eighteen SD rats with the weight of 100g were randomly divided into 3 groups:group A (standard diet group),group B(high-fat diet group) and group C (treadmill-trained group with high-fat diet).Training (1 hour/d) initiated at the same time as the HF diet was fed.After being raised for 6 weeks,the rats was euthanized and weighed up.Blood samples were taken and the levels of serum lipid were detected.The levels of serum endotoxin were detected by enzyme-linked immunoadsorbent assay.The membrane potentials of isolated mitochondrion were detected by flow cytometry instrument and the morphologic changes in mitochondria in liver were observed by electronic microscopy.Results In group B,the levels of endotoxin increased significantly(2.916 ± 0.761 rs 5.454 ± 1.254,t =-4.236,P ＜ 0.05),and the liver mitochondrial density and membrane potential also increased significantly compared with group A after 6 weeks (4.330 ±0.501 vs 3.507 ±0.532,t =2.759,P ＜0.05;l.660 ±0.202 vs 0.473 ±0.064,t =13.712,P ＜0.05).But there was no markedly different in serum endotoxin between group B and group C (4.972 ± 1.757 vs 5.454 ± 1.254,t =-0.547,P ＞ 0.05).Compared with group B,the liver mitochondrial density of group C decreased significantly (4.330±0.501 vs 3.581 ±0.188,t =3.426,P ＜ 0.05).The mitochondrial ultrastructurctural changes in each group were not obvious.Conclusions The rats fed with high-fat diet for 6 weeks can reach the state of metabolic endotoxemia.The increasing levels of the liver mitochondrial membrane potential caused by metabolic endotoxin may affect the happening and development of other diseases in the future.Concurrent exercise can not decrease the level of endotoxin.It also shows that metabolic disease caused by high-fat diet should be prevented by moderation in eating and drinking.
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Objective To investigate the protective effect and possible mechanisms of continuous infusion of norepinephrine in kidney of septic rats in the early stage.Methods Thirty healthy male SD rats of SPF level were randomly divided into five groups.Rats in control group were given intraperitoneal injection of saline and began a continuous infusion of saline (1 ml/h).Rats in LPS group and the intervention group (low-dose,medium-dose and high-dose norepinephrine group) were given intraperitoneal injection of LPS 10 mg/kg.LPS group began a continuous infusion of saline (1 ml/h) while low,medium and high dose groups began continuous infusion of different norepinephrine solution [(0.06,0.3,0.6 μg/(kg·min)].Rats were sacrificed after 24 hours infusion.We detected serum inflammatory cytokines [tumor necrosis factor (TNF)-α,interleukin(IL)-1β,IL-6 and IL-10] in 2 h and 6 h by ELISA.Rat serum CRP,Cr and BUN,swelling and membrane potential of kidney mitochondria and oxidative stress-related indicators were tested in 24 h.We also observed renal pathologic changes by electronic microscopy and biopsy.Results Compared with the control group,serum levels of TNF-α [(2 203.3 ± 1 028.7) pg/ml],IL-1β [(2 214.5 ±457.0) pg/ml],IL-6 [(7784.7 ±248.2) pg/ml] and IL-10 [(1 076.1 ±368.4) pg/ml] were statistically higher in LPS group in 2 h (P ＜ 0.05) ; CRP [(0.35 ± 0.24) mg/L],Cr [(30.8 ± 11.5) μ mol/L],BUN [(7.7 ± 1.8) mmol/L],NO [(1 057.4 ± 172.9) μmol/gprot] were statistically higher (P ＜ 0.01),membrane potential of kidney mitochondria (0.0464 ±0.018 5) decreased statistically (P ＜0.01).Compared with LPS group,serum levels of TNF-αt [(506.8 ±301.7) pg/ml],IL-lβ [(415.6 ± 178.0) pg/ml],and IL-10 [(381.7 ± 171.0) pg/ml] significantly decreased in low-dose group in 2 h (P ＜0.05),BUN [(5.2 ± 1.4)mmol/L] decreased (P ＜ 0.05),mitochondrial membrane potential (0.347 4 ± 0.152 6) increased in 24 h (P ＜ 0.05) ; serum levels of TNF-α [(323.9 ± 227.9) pg/ml],IL-1 β [(700.0 ± 246.2) pg/ml],and IL-10[(282.6 ± 134.4) pg/ml] significantly decreased statistically in medium-dose group in 2 h (P ＜0.05),CRP [(0.17 ± 0.08) mg/L] decreased statistically (P ＜ 0.01),mitochondrial membrane potential (0.377 5 ±0.143 7) increased in 24 h (P ＜0.05) ;serum levels of TNF-α [(378.7 ±89.8) pg/ml],IL-1β [(945.7 ±264.4) pg/ml] significantly decreased in high-dose group in 2 h (P ＜0.05),CRP [(0.19 ±0.12) mg/L] and Cr [(23.2 ±3.4) μmol/L] decreased in 24 h (P ＜0.05).Mitochondrial matrix coated fuzzy,vacuoles and coagulation were found in LPS group by electronic microscopy examination.Interstitial edema,monocyte-macrophage infiltration,glomerular shrinkage,tubular epithelial cell swelling,empty bubble degeneration were found in LPS group by microscopy examination.Pathological damage was alleviated in mediumdose group.Conclusion Continuous infusion of norepinephrine plays a protective role on renal function in rats with sepsis in the early stage.The intervention protect rat kidney by reducing levels of inflammatory cytokines,oxidative stress and mitochondrial damage.