ABSTRACT
Objective To determine the relationship between brain injury and cerebral glucose metabolism in rat model of cardiac arrest. Methods Asphyxia-induced cardiac arrest model was established. Forty-two male Wistar rats were randomly assigned to sham or experimental groups. Rats in the CA4,CA6 and CA8 group were treated with cardiopulmonary resuscitation(CPR) 4 min, 6 min and 8 min after cardiac arrest, respectively. The maximum standardized uptake value (SUVmax) of glucose was detected by PET, and neural defi cit score (NDS) were evaluated at 24 h and 72 h after ROSC. The numbers of injured neurons and apoptotic cells and the protein level of hexokinase I (HXK I) were measured at 72 h after ROSC. Results SUVmax, NDS and the level of HXK I were all decreased after ROSC, and interestingly, this declination of these markers was correlated with the prolongation of the duration of CA, the longer duration of CA the more declination of these biomarkers. Accordingly, the number of injured neurons and apoptotic cells increased were correlated with duration of CA, and thus CA8 group had greater numbers of those cells than CA6 group and CA4 group (P<0.05),and CA6 group had greater numbers of those cells than CA4 group(P<0.05). In addition, the SUVmaxwas positively correlated with NDS(P<0.05), and negatively correlated with the numbers of injured neurons and apoptotic index(P<0.05). Conclusions The degree of brain injury is associated with cerebral glucose metabolism, and PET may become a novel method to assess the severity of brain damage after CA.
ABSTRACT
Objective To observe the effects of mild hypothermia on the myocardial mitochondrial injury induced by oxidative stress after restoration of spontaneous circulation (ROSC) in rat of cardiac arrest model.Methods Eighteen male Wistar rats were randomly (raudom number) divided into normal temperature group and mild hypothermia group after ROSC.Ultrasound was used to measure the left ventricular ejection fraction (EF),shortening fraction (FS) and stroke volume (SV).The levels of glutathione (GSH),malondialdehyde (MDA) and adenosine triphosphate (ATP) in myocardium were detected.The ultramicroscopic structure of myocardial mitochondria was observed under transmission electron microscope at 4 h after ROSC.Results There were no significant differences in basic life support (BLS) time,dosage of epinephrine and number of defibrillation attempt between two groups (P ＞ 0.05).The concentrations of GSH and ATP in myocardium of rats in hypothermia group were significantly higher than those in normal temperature group,while the level of MDA was significantly lower in hypothermia group than that in normal temperature group.Echocardiographic findings showed that hypothermia could significantly improve the EF,FS and SV after ROSC.The hypothermia decreased the myocardial mitochondria injury rather than normothermia [mitochondrial injury score:(0.21-±0.04) vs.(0.42 ±0.08),P ＜ 0.05].Conclusions In this model,mild hypothermia can decrease myocardial oxidative stress injury,improving the cardiac function after ROSC.
ABSTRACT
Objective To establish the cardiac arrest-cardiopulmonary resuscitation model in rats, and to observe the effect of mild hypothermia on autophagy in hippocampal CA1 neurons after ROSC.Methods A total of 36 Wistar rats were randomly divided into two groups: normal temperature treatment group(NT group) and mild hypothermia treatment group(HT group).To establish the cardiac arrest-cardiopulmonary resuscitation(CA-CPR) model in rats by epicardial electrical stimulation induced ventricular fibrillation, and to sacrifice 3 animals in each group to obtain the brain cortex in 2nd and 4th hours after ROSC in order to observe the expression of p-AMPK by electron microscope and LC3 granules through Western blot.The neurological deficit score(NDS) was assessed in 24、48、72 hours respectively after ROSC.To sacrifice the animals so as to take the cerebrum in 72 hours after ROSC, then calculate the apoptotic index of the hippocampal CA1 neurons, which were dyed through TUNEL method.Results The expression of p-AMPK、Beclin-1 and LC3-Ⅱ/LC3-Ⅰratio in Normothermia group were all lower than the Mild hypothermia group(P<0.05), the neurons plasma of hippocampal CA1 area in the Hypothermia group demonstrated obvious LC3 granules formation, the NDS score of the Normothermia group and the Mild hypothermia group in ROSC24h、ROSC48h、ROSC72h were 320vs205、285vs140、266vs120, respectively.The apoptotic index of the hippocampal CA1 area in the Normothemia group in ROSC72h was higher than the Mild hypothermia group,(P<0.05).Conclusions Mild hypothermia after cardiopulmonary resuscitation promotes autophagy of the hippocampal CA1 area neurons in rats and reduce neuronal apoptosis.
ABSTRACT
AIM: To explore the effect and possible mechanism of type 2 innate lymphoid cell (ILC2) on the development of chronic renal failure (CRF).METHODS: The patients with chronic renal failure (n=36) in the Fist Affiliated Hospital of Sun Yat-sen University from March 2016 to December 2016 were selected, and 32 healthy persons in the same period were enrolled in the study for control.The proportion of ILC2 in the PBMC of CRF patients and healthy controls was detected by flow cytometry, IL-13 concentration in the plasma was measured by ELISA.The isolated PBMCs from the patients and healthy persons were divided into 3 groups (control group, cytokine group, intervention group) and cultured in vitro for 3 days, respespestively, then IL-13 concentration was measured by ELISA.The protein levels of phosphorylated signal transducers and activators of transcription 6 (p-STAT6) in the PBMC of healthy controls before stimulation and after stimulation for 15 min, 30 min, 1 h, 2 h were determined by Western blot.RESULTS: The proportion of ILC2 in the PBMC and the plasma IL-13 concentration of CRF patients was higher than that in the healthy controls (P<0.05).In the culture supernatant in vitro, IL-13 concentration in the 3 subgroups of CRF patients (control group, cytokine group, intervention group) were all higher than that in the healthy controls (P<0.05), both the 2 groups showed a trend that the active IL-13 concentration in cytokine group was higher than that in control group, and that in intervention group was lower than that in cytokine group.The protein levels of p-STAT6 in cytokine stimulated-PBMC with a time dependent manner.CONCLUSION: The percentage of ILC2 in the PBMC is elevated in CRF patients.Furthermore, the ILC2 secret large amount of IL-13 to mediate the polarization of Th2 cells to regulate immunity through activating p-STAT6.
ABSTRACT
Objective To compare clinical data of the death in different intensive care unit,in order to provide the medical strategies for patients in EICU.Methods The clinical data of lethal cases from January 1,2013 to December 31,2014 in EICU,SICU and MICU of the First Affiliated Hospital of Sun Yat-sen University were compared.EICU (252 cases),SICU (93 cases) and MICU (80 cases) were enrolled.The demographics of each patient,clinical condition such as critical score (APACHE Ⅱ score),length of stay,overall costs,and the patient families' different opinions to the treatment in each ICU were analyzed.The data was analyzed with SPSS 13.0 software,averaged value was presented as mean ± standard and the non-normal distributions were expressed as median (25％,75％).The one-way analysis of variance was followed by the Tukey post hoc test for pairwise comparisons and chi-square test was used for comparison of percentage between two groups.Results Two hundred and fifty-two cases in EICU had gender ration of 148/96 (male/female),92 cases in SICU 68/24,80 cases in MICU 56/24.Ages of the fatal were EICU 72 ± 17 years,SICU 56 ± 17 years,and MICU 63 ± 20 years,respectively.Age of the fatal in EICU was significantly older than that of the SICU (P ＜ 0.01) and the MICU (P ＜ 0.01).APACHE Ⅱscores were 33 ± 8 in EICU,34 ± 10 in SICU,29 ± 10 in MICU,respectively.The severity scores in EICU patients were higher than those in MICU patients and SICU patients (P =0.01 and 0.021).Lengths of stay were 2 days (1,46) in EICU,14 days (1,84) in SICU,12 days (1,77) in MICU,respectively.EICU hospitalization time was significantly shorter than that of SICU (P ＜ 0.01) and the MICU (P ＜ 0.01).Total costs of hospitalization were 9 777 yuan (400,164 126) yuan in EICU,100 628 yuan (13 639,964 783) yuan in SICU,119 463 yuan (5 650,590 903) yuan in MICU,and that in EICU was significantly less than the total cost of hospitalization in SICU (P ＜ 0.01) and in MICU (P ＜ 0.01).The opinion of patient families was proposed to give up treatment associated with 165 dead cases in EICU,18 death cases in SICU and 20 dead cases in MICU,and the rate of discontinuous treatment in EICU patients was significantly greater than that in SICU (P ＜ 0.01) and in MICU (P ＜ 0.01).There were no significant differences in invasive procedures,invasive hemodynamic monitoring,mechanical ventilation,blood purification and deep vein puncture among three groups.The 5 leading causes in EICU were severe sepsis,stroke,sudden cardiac arrest,acute myocardial infarction and advanced malignancy.Conclusions The death of patients were due to advanced age with severe disease,poor prognosis,and the request of patient family members to give up treatment.The 5 leading causes were severe sepsis,stroke,sudden cardiac arrest,acute myocardial infarction and advanced tumors suggesting the establishment of corresponding treatment scheme to be made and preparation of abundant medical resources to be ready.Timely communication with the patients' families and let them participate in end-stage treatment decisions was the best strategies to improve the successful rate of treating severe patients and use EICU resource effectively.
ABSTRACT
Objective To investigate the neurons autophagy and apoptosis after cerebral ischemia reperfusion (I/R) injury in mice,and to explore the effect of mild hypothermia on neurons autophagy and apoptosis.Methods The global cerebral ischemia-reperfusion injury model in C57 mice was established with carotid artery ligation method.Ninety-six C57 mice were randomly (random number) divided into 8 groups (n =12 in each group),namely control group (C0),sham group,normal body temperature group (NT,37 ℃) after I/R 6 h (C6 h),normal body temperature group after I/R 12 h (C12 h),normal body temperature group after I/R 72 h (C72 h),mild hypothermia group (MH,34 ℃) after I/R6 h (C6 h +MH),mild hypothermia group after I/R12 h (C12 h + MH),mild hypothermia group after I/R 72 h (C72 h + MH).The protein expressions of Sirt1,P-FoxO1,Rab7,P53 and autophagy related genes such as Beclin1,LC3 were detected by Western blot at given intervals.The LC3 granules were assayed by immunofluorescence.The neurons apoptosis was detected by TUNEL.The software of SPSS13.0 was used for statistical analysis.Measurement data was expressed with mean ± SD,and comparison between two groups was carried out with Student's t test,One-way ANOVA was used for comparisons among groups,and P ＜ 0.05 was considered statistically significant.Results The global cerebral ischemia-reperfusion injury model in C57 mice was established successfully with bilateral carotid arteries ligation method.Compared with the control group,the protein expressions of Sirt1,P-FoxO1,Rab7,Beclin1 and LC3 Ⅱ / Ⅰ were gradually reduced,especially at 12 h after I/R in NT group (P ＜ 0.05),while the expression of P53 was obviously increased (P ＜0.05).In MH group,the expressions of Sirt1,P-FoxO1,Rab7,Beclin1,LC3 Ⅱ / Ⅰ were higher than those in NT group (P ＜ 0.05).And the expression of P53 was lower than that in NT group (P ＜0.05).Immuno-fluorescence test showed that compared with the control group,the LC3 particles of neurons cells were decreased significantly in group C12 (at 12 h after I/R 6.0 ± 1.5 vs.18.1 ±2.5,P ＜0.05).Nevertheless,LC3 particles were increased in MH group compared with NT group (36.1 ± 4.5 vs.6.0 ± 1.5,P ＜ 0.05).The results of TUNEL test showed that compared with the control group,neurons cells apoptosis were significantly increased in C72 group (at 72 h after I/R,54.8％ ±7.5％ vs.5.5％ ±1.2％,P ＜ 0.05).However,compared with NT group,neurons apoptosis were decreased in MH group (28.8％ ±4.5％ vs.54.8％ ±7.5％,P＜0.05).Conclusions The neuron autophagy was significantly reduced and apoptosis was significantly increased after ischemia reperfusion injury (I/R) in mice.However,mild hypothermia could increase the expression of Sirt1,FoxO1,beclin1 and LC3,so as to promote neurons autophagy and reduce apoptosis,which would provide therapy target for neurons injury after hypoxia and provide soundly theoretical basis for mild hypothermia for clinical application.
ABSTRACT
Objective To investigate the neurons autophagy and apoptosis after cerebral ischemia reperfusion (I/R) injury in mice,and to explore the effect of mild hypothermia on neurons autophagy and apoptosis.Methods The global cerebral ischemia-reperfusion injury model in C57 mice was established with carotid artery ligation method.Ninety-six C57 mice were randomly (random number) divided into 8 groups (n =12 in each group),namely control group (C0),sham group,normal body temperature group (NT,37 ℃) after I/R 6 h (C6 h),normal body temperature group after I/R 12 h (C12 h),normal body temperature group after I/R 72 h (C72 h),mild hypothermia group (MH,34 ℃) after I/R6 h (C6 h +MH),mild hypothermia group after I/R12 h (C12 h + MH),mild hypothermia group after I/R 72 h (C72 h + MH).The protein expressions of Sirt1,P-FoxO1,Rab7,P53 and autophagy related genes such as Beclin1,LC3 were detected by Western blot at given intervals.The LC3 granules were assayed by immunofluorescence.The neurons apoptosis was detected by TUNEL.The software of SPSS13.0 was used for statistical analysis.Measurement data was expressed with mean ± SD,and comparison between two groups was carried out with Student's t test,One-way ANOVA was used for comparisons among groups,and P ＜ 0.05 was considered statistically significant.Results The global cerebral ischemia-reperfusion injury model in C57 mice was established successfully with bilateral carotid arteries ligation method.Compared with the control group,the protein expressions of Sirt1,P-FoxO1,Rab7,Beclin1 and LC3 Ⅱ / Ⅰ were gradually reduced,especially at 12 h after I/R in NT group (P ＜ 0.05),while the expression of P53 was obviously increased (P ＜0.05).In MH group,the expressions of Sirt1,P-FoxO1,Rab7,Beclin1,LC3 Ⅱ / Ⅰ were higher than those in NT group (P ＜ 0.05).And the expression of P53 was lower than that in NT group (P ＜0.05).Immuno-fluorescence test showed that compared with the control group,the LC3 particles of neurons cells were decreased significantly in group C12 (at 12 h after I/R 6.0 ± 1.5 vs.18.1 ±2.5,P ＜0.05).Nevertheless,LC3 particles were increased in MH group compared with NT group (36.1 ± 4.5 vs.6.0 ± 1.5,P ＜ 0.05).The results of TUNEL test showed that compared with the control group,neurons cells apoptosis were significantly increased in C72 group (at 72 h after I/R,54.8％ ±7.5％ vs.5.5％ ±1.2％,P ＜ 0.05).However,compared with NT group,neurons apoptosis were decreased in MH group (28.8％ ±4.5％ vs.54.8％ ±7.5％,P＜0.05).Conclusions The neuron autophagy was significantly reduced and apoptosis was significantly increased after ischemia reperfusion injury (I/R) in mice.However,mild hypothermia could increase the expression of Sirt1,FoxO1,beclin1 and LC3,so as to promote neurons autophagy and reduce apoptosis,which would provide therapy target for neurons injury after hypoxia and provide soundly theoretical basis for mild hypothermia for clinical application.
ABSTRACT
Objective To evaluate the effect of extracorporeal cardiopulmonary resuscitation (ECPR) and conventional cardiopulmonary resuscitation (CCPR) on survival and neurological function in adult patients with cardiac arrest.Methods The PubMed and Web of Science were searched to collect relevant literature from Jan 1980 to Nov 2015,and two reviewers strictly distinguished the studies,assessed the quality of studies and picked up the valuable data for statistical analysis by using RevMan 5.0.Results A total of 8 studies involving 27 18 patients were included in our review.Of them,462 patients were treated with ECPR and 2 256 patients were cared with CCPR.The meta analysis showed that the survival discharge rate (OR =2.92,95％ CI:2.24-3.81,P ＜ 0.01),long-term survival rate (OR =2.97,95％ CI:2.11-4.19,P＜0.01) and neurological function status (OR=3.50,95％CI:2.36-5.81,P＜ 0.01) of ECPR (n =182) were better than those of CCPR (n =182).In 4 studies,propensity score matching was used to minimize bias and heterogeneity.The meta analysis also showed that the rate of ROSC,survival discharge rate,long-term survival rate and neurological function status in ECPR were superior over CCPR.Conclusions ECPR would be the excellent measures to improve ROSC rate,survival discharge rate,long-term survival rate and neurological outcome in adult victims with cardiac arrest.
ABSTRACT
Objective To investigate the clinical features and risk factors of recurrent communityacquired pneumonia (CAP) in elderly patients.Methods Clinical data of elderly CAP patients admitted to the first affiliated hospital of Sun Yat-Sen university from January 2012 to December 2014 were analyzed retrospectively,including 48 cases of recurrent CAP and 299 cases of non-recurrent CAP.The clinical features were compared between two groups,and the risk factors of recurrent CAP were assessed with logistic regression analysis.Results No significant differences were observed between the two groups in age,sex,smoking,drinking,average hospital stays and total hospitalization expenses (all P ＞ 0.05).In the recurrent pneumonia group,the chronic obstructive pulmonary diseases [54.2％ (26/48) vs.35.5％ (106/299),χ2 =6.146,P=0.013],diabetes [41.7％ (20/48) vs.19.7％ (59/299),χ2 =11.317,P=0.001],the clinical symptoms include cough [79.2％ (38/48) vs.58.5％ (175/299),χ2 =7.432,P =0.006],sputum [70.8％ (34/48) vs.54.5％ (163/299),χ2 =4.488,P =0.034],fever [64.4％ (31/48) vs.40.8％(122/299),χ2 =9.488,P =0.002],poor appetite [52.1％ (25/48) vs.33.1％ (99/299),χ2 =6.483,P =0.011],choking cough [8.3％ (4/48) vs.2.7％ (8/299),χ2 =3.966,P =0.046],poor spirit [45.8％ (22/48) vs.28.4％ (85/299),χ2 =5.875,P =0.015],bilateral lung lesions [41.7％ (20/48)vs.28.8％ (68/299),χ2 =7.825,P =0.005],the complication of respiratory failure [31.3 ％ (15/48) vs.14.7％ (44/299),χ2 =8.012,P =0.005] were higher than those in non-recurrent group.Logistic regression revealed that chronic obstructive pulmonary disease (OR =2.607,95％ CI:1.268-5.358,P =0.009),diabetes (OR =2.948,95％ CI:1.457-5.966,P =0.003) and the history of respiratory failure (0R=2.778,95％CI:1.264-6.108,P=0.011) were risk factors of recurrent CAP.Conclusions Elderly patients with recurrent CAP has a higher rate of cough,sputum and fever,and the scope of lung lesions is broader.Chronic obstructive pulmonary disease,diabetes and the history of respiratory failure are the independent risk factors for recurrent CAP in the elderly.
ABSTRACT
Objective To explore the effects of estrogen on oxidative stress of the lung tissue induced by acute paraquat (PQ) poisoning. Methods Thirty-two male adult New Zealand rabbits were randomly divided into model group and estrogen intervention group, 16 rabbits in each group. The model of lung injury induced by PQ poisoning was reproduced by feeding 16 mg/kg of 20% PQ through gastric tube. The rabbits in estrogen intervention group received intravenous infusion of 5 mg/kg estrogen after PQ challenge for 7 days, and the rabbits in model group received an equal volume of normal saline. Three rabbits in each group were sacrificed at 1, 2 and 3 days respectively after exposure. The lung tissue was harvested, the levels of reactive oxygen species (ROS) was determined by 2',7'-dichlorofluorescin diacetate (DCFH-DA), malondialdehyde (MDA) was determined by thiobarbituric acid (TBA), the mRNA expression of manganese-containing superoxide dismutase (MnSOD) was determined by reverse transcription-polymerase chain reaction (RT-PCR), and adenosine triphosphatase (ATP) content in mitochondrion was determined by enzyme linked immunosorbent assay (ELISA). The pathological changes in lung were observed under light microscopy using hematoxylin and eosin (HE) staining, and the lung injury was evaluated with lung injury score. Results The contents of ROS and MDA in lung within 3 days after PQ poisoning were gradually increased, and MnSOD mRNA expression and ATP content were gradually decreased. Estrogen intervention could significantly reduce the production of ROS and MDA after PQ poisoning [3-day ROS (fluorescence intensity): 161.05±30.04 vs. 188.30±31.80, 3-day MDA (mmol/L): 98.71±0.92 vs. 122.12±1.24], up-regulate MnSOD mRNA expression (integral A value: 3.05±0.90 vs. 1.22±0.24), and increase ATP content in mitochondrion (ng/L: 3.75±0.92 vs. 2.28±0.29) with statistically significant differences (all P < 0.01). In lung tissue after PQ poisoning, congestion, edema, focal pulmonary consolidation, pulmonary interstitial and alveolar space were infiltrated by a large number of neutrophil, alveolar interval were thickened obviously and the above phenomenon were most serious at 3 days after poisoning as shown under optical microscope. Estrogen intervention could significantly improve lung injury as compared with that of model group, and the lung injury score at 3 days was significantly lower than that of model group (11.8±0.7 vs. 13.5±1.0, P < 0.01). Conclusions The oxidative stress indicators in the lung tissue after PQ poisoning were obviously abnormal, the pathological damage was serious with time dependence. The administration of estrogen can reduce acute lung injury after PQ poisoning by reducing the oxidative stress.
ABSTRACT
AIM:To investigate the neuroprotective effect of hydrogen sulfide ( H2 S) after cardiopulmonary re-suscitation in rats with cardiac arrest ( CA) , and to explore the effects of H2 S on neuron autophagy.METHODS:The CA model was established through asphyxia.Male Wistar rats were randomly divided into sham group, model group and NaHS group.The levels of beclin-1 and LC3 II/I were measured by Western blot at 2 h, 4 h, 12 h and 24 h after the restoration of spontaneous circulation (ROSC).At 12 h after ROSC, the formation of autophagic vacuole with LC3 dots was deter-mined by immunohistochemical ( IHC) method.The phenomenon of neuron autophagy was observed under transmission electron microscope.The numbers of apoptotic neurons were counted by TUNEL staining at 72 h after ROSC.The neurolo-gic deficit score ( NDS) was used to evaluate the neurologic function after ROSC.RESULTS: The level of beclin-1 was gradually increased in model group, but it was increased and then gradually recovered in NaHS group ( P<0.05 ) .The conversion of LC3 II in the cerebral cortex was the same as beclin-1.The results of IHC showed that LC3-positive nuclei in model group were more than those in NaHS group ( P<0.05) .The number of autophagic vacuole in model group was more than that in NaHS group (P<0.05).The number of the TUNEL-positive cells in model group was more than that in NaHS group (P<0.05).The NDS of the animals in NaHS group after ROSC was lower than that in model group(P<0.05). CONCLUSION:H2 S inhibits neuronal autophagy, decreases apoptosis and improves neurologic function in CA rats after ROSC.
ABSTRACT
Objective To evaluate the effects of Ulinastatin (UTI) on the expressions of TNF-α,IL-6 and neurons apoptosis in cerebral cortex of rats after cardiopulmonary resuscitation (CPR).Methods Thirty-six healthy male adult Wistar rats were induced ventricular fibrillation untreated for 7 min and then received CPR.The animals were infused UTI 100 000 U/kg or phosphate-buffered solution (PBS) at once after ROSC.At 2,4 and 8 h after ROSC,cerebral cortex were removed to determine the mRNA expressions and levels of TNF-α protein and IL-6 protein,the translocation ratio of NF-κB p65 from cytoplasm to nucleus and the apoptotic neurons.Results The plasma levels of TNF-α (ng/mL) in animals of UTI group were (17.7 ± 1.4),(21.9 ± 2.1) and (17.1 ± 0.6) at 2,4 and 8 h after ROSC respectively,and significantly lower than those in PBS group at the given intervals.Mean while,the levels of IL-6 (ng/mL) were (208.9 ± 14.1),(281.5 ±25.9) and (251.8 ± 15.3) at 2,4 and 8 h after ROSC respectivèly in animals of UTI group,and lower than those in PBS group.The expressions of TNF-α mRNA and IL-6 mRNA and protein levels of TNF-α and IL-6 in UTI group were both lower than those in PBS group at given intervals,respectively.The translocation ratio of NF-κB p65 from plasma to nucleus in PBS group at each given interval after ROSC was significantly higher than that in UTI group.The number of viable neurons in cerebral cortex in UTI group was higher than that in PBS group,while the number apoptosis neurons was fewer in UTI group.Conclusions UTI attenuated the general inflammatory response after ROSC in rat,decreased the activation of NF-κB pathway,and subsequently attenuated the expression of TNF-α and IL-6,and finally decreased the neurons apoptosis.
ABSTRACT
[ ABSTRACT] AIM:To investigate the effects of induced pluripotent stem cells-derived mesenchymal stem cells ( iPSC-MSCs) on cobalt chloride ( CoCl2 )-induced injuries of PC12 cells and its possible mechanism.METHODS:PC12 cells were exposed to CoCl2 to set up a chemical-induced cellular injury model and were cocultured with iPSC-MSCs.The cell viability was tested by CCK-8 assay.The apoptosis was measured by flow cytometry using Annexin V/PI staining.The mitochondrial membrane potential (MMP) was analyzed by flow cytometry using JC-1 staining.Immunofluorescence was employed to observe mitochondrial transfer from iPSC-MSCs to PC12 cells.RESULTS: Apoptosis of PC12 cells was in-creased and MMP of PC12 cells was decreased after exposed to CoCl2 at concentration of 400μmol/L for 24 h.Coculture of PC12 cells with iPSC-MSCs reduced the apoptosis and recovered the MMP of the PC12 cells.Tunneling nanotubes were formed between iPSC-MSCs and PC12 cells, through which the iPSC-MSCs transferred the mitochondria to the PC12 cells. CONCLUSION:iPSC-MSCs protect PC12 cells from CoCl2-induced injuries, which may be associated with the mitochon-drial transfer from iPSC-MSCs to PC12 cells.
ABSTRACT
AIM: To investigate the effect of cobalt chloride (CoCl2) on the apoptosis of neural stem cells (NSCs) and the expression of microRNA-26a (miR-26a) in vitro, and to explore the mechanisms of NSC apoptosis in-duced by CoCl 2 .METHODS:NSCs were exposed to CoCl 2 at different doses (200~600μmol/L) for 24 h.The cell via-bility and apoptosis were measured by CCK-8 assay and TUNEL method.The expression of miR-26a-3p, miR-26a-5p, GSK-3β, caspase-3, Bcl-2 and Bax was examined by real-time PCR.The protein levels of Bcl-2 and Bax were detected by Western blotting .RESULTS: The cell viability was inhibited and the apoptosis of NSCs was increased significantly by CoCl2 in a dose-dependent manner (P<0.05).CoCl2 at concentration of 400μmol/L for 24 h was used to induce apopto-sis and the expression of miR-26a was down-regulated compared with control (P<0.05).Exposure to CoCl2 at concentra-tion of 400μmol/L up-regulated the expression of GSK-3β, caspase-3 and Bax , down-regulated the expression of Bcl-2 and Bcl-2/Bax (P<0.05).CONCLUSION:CoCl2 at concentration of 400μmol/L induces the apoptosis of NSCs obviously . CoCl2 may induce the NSC apoptosis by mitochondrial apoptotic pathway .Declining miR-26a may be related to NSC apopto-sis.
ABSTRACT
Objective To study the effect of neuronal Nogo-66 receptor (NgR1) antagonist,soluble Nogo-66 receptor (sNgR1-Fc),on promoting the endogenous neural precursor cells (NPCs) differentiating into neurons in order to clarify the mechanism.Methods The cortical infarction was induced by photochemistry,named photothrombotic cortical injury (PCI).Twelve Sprague Dawley rats were randomly divided (random number) into three groups:Sham-operated group,PBS group,and sNgR1-Fc group.PBS (PBS group) or sNgR1-Fc (sNgR1-Fc group) was injected into the lateral ventricle of brain with a minipump.BrdU (Bromodeoxyuridine) was injected into the peritoneal cavity 4-6 days after PCI.The subdentate gyrus zone (SGZ) of brain from sacrificed rat was harvested for Immunohistochemistry to observe the ratio of NeuN +/BrdU + cells 35 days after PCI.Proteins including Nestin、Notch1 and Mash1 were detected by Western Blot.Results The cortical infarction in rat was successfully induced by photochemistry.Thirty-five days after PCI,the BrdU + cells number and theratio of NeuN +/BrdU + in the SGZ of the ipsilateral cerebrum hemisphere with PCI were significantly higher in sNgR1-Fc group than those in PBS group (P ＜ 0.05).The levels of Notch1,Mash1 and Neuro D in the sNgR1-Fc group were significantly higher than those in the PBS group (P ＜ 0.05),which were significantly higher than those in the Sham-operated group.Conclusions sNgR1-Fc could promote the endogenous NPCs differentiating into neurons in a cortical infarction model.The mechanisms may be attributed to the Notch/bHLH (proneural basic helix-loop-helix genes) signaling way.
ABSTRACT
Objective To evaluate the effect of percutaneous coronary intervention (PCI) or thrombolysis,in patients with return of spontaneous circulation (ROSC) after out-of-hospital cardiac arrest (OHCA),in the presence of ST-elevation myocardial infarction (STEMI).We demonstrated the benefits of the two therapies on ROSC patients in hospital discharge and neurological recovery,and clarified the importance of ROSC,so as to guide the treatments for OHCA in the presence of STEMI.Methods It was performed a meta-analysis of clinical studies located in PUBMED and MEDLINE databases from January 1995 to October 2011.OHCA patients with ROSC were as our study objects,the hospital discharge and neurological recovery rates,of patients with and without PCI or thrombolysis,were assessed in patients with ROSC after OHCA in the presence of STEMI.In the same Cohort Study,between received and rejected PCI,or between received and rejected thrombolysis in OHCA patients with ROSC as treated group and control group,using Review Manager 5.1 software to analyze,respectively.Furthermore,we also compared the differences in hospital discharge and neurological recovery rates between patient groups who received PCI or thrombolysis by Pearson x2 analysis.Results The meta-analysis showed that the rate of hospital discharge improved with both PCI (odds ratio [OR],1.65 ; 95％ confidence interval [CI],1.05-2.59,P ＜ 0.01)and thrombolysis (OR,2.03 ; 95％ CI,1.24-3.34,P ＜ 0.01) in patients with ROSC after OHCA,in the presence of STEMI.We also found that there were not significant differences between with PCI and with thrombolysis in the rate of hospital discharge (63.00％ vs.65.19％,P =0.548) and neurological recovery (88.62％ vs.91.25％,P =0.351) for the patients with ROSC after OHCA (P ＞0.05).Conclusions In patients with ROSC after OHCA in the presence of STEMI,both PCI and thrombolysis improved hospital discharge rates.Furthermore,there were similar efficacy in hospital discharge and neurological recovery rates between with PCI and with thrombolysis.
ABSTRACT
Objective To investigate the therapeutic effects of enhanced external counterpulsation (EECP) on cerebral edema and cerebral blood flow perfusion with MRI following cardiac arrest (CA) and on successful return of spontaneous circulation (ROSC) by cardiopulmonary resuscitation (CPR) in dogs.Methods Sixteen beagle dogs were induced CA with alternating current on epicardium,then were randomly (random number) divided into the EECP and control group after successful ROSC.MR scanning brain of all animals was carried out by diffusion-weighted imaging (DWI) and perfusion weighted imaging (PWI) before CA and on the first,second and third days after ROSC.Blood pressure,right common carotid artery flow,and intracranial microcirculation perfusion were measured.Results There were no significant differences in mean artery pressure at all intervals between two groups (P ＞ 0.05).There was significant increase in right common carotid artery blood flow and intracranial microcirculation of dogs in EECP group compared with the control group (P ＜ 0.05).Apparent diffusion coefficients (ADC) of water molecule on the first and third days after ROSC were significantly higher in the EECP group than those in the control group (P ＜ 0.05).Ratios of post-ROSC relative cerebral blood flow (RCBF) /original cerebral blood flow were higher in the EECP group than those in the control group on the first,second and third days after ROSC (P ＜ 0.05).Conclusions EECP treatment could improve cerebral blood flow perfusion and relieve ischemic cerebral edema,alleviating brain injury in dogs following CA and successful ROSC.
ABSTRACT
Objective To observe the protective effects of soluble Nogo-66 receptor (NgR1 )antagonist (sNgR1-Fc) on cortical axons after cortical infarction in rats,and to study the phenomenon and molecular mechanism of its protective effects on and regeneration of axons.Methods The cortical infarction was induced by photochemistry,termed photothrombotic cortical injury (PCI).Fifteen Sprague Dawley rats were randomly divided into three groups:Sham-operated group,PBS (phosphate buffered solution) group,and s-NgR1-Fc group.In PBS group,PBS was injected into the lateral ventricle of rats; and in sNgR1-Fc group,sNgR1-Fc was injected instead of PBS. The ipsilateral cortex with lesion was harvested for histomorphometry and transmission electron microscope observation 7 days after PCI. Proteins including GTP-RhoA,p-JNK,p-c-JUN and p-ATF-2 were detected by Western blot,as well as Total-J and Total-RhoA.Results The cortical infarction in rats was successfully induced by photochemistry.Compared with sham-operated group,the pathological changes in PBS groups were more serious,including extensive edema or disappearance of axoplasm of fiber without medulla sheath involved and extensive thickening or layer derangement in axoplasm of fiber with medulla sheath involved.These changes were improved significantly after sNgR1-Fc treatment.The levels of GTP-RhoA,p-JNK1,p-JNK2,p-c-JUN and p-ATF-2 in the PBS group were significantly higher than those in the sham-operation group ( P ＜ 0.05 ),whereas the levels of Total-RhoA,Total-JNKl and Total-JNK2 were not different significantly between these two groups (P ＞0.05 ).The sNgR1-Fc treatment up-regulated the levels of these proteins ( P ＜ 0.05 ).Conclusions There is pathological change in axon induced by cerebral hypoxia-ischemia for a long period after cortical infarction.The mechanisms may be associated with RhoA/ROCK/JNK/c-Jun signal way,which is activated by ischemia injury and related to the inhibition of regeneration in axon.Our study shows that NgR1-Fc may inhibit this pathway significantly,and then promote the regeneration of axon partially.
ABSTRACT
Objective To investigate whether Ulinastatin (UTI) would minimize the systemic inflammatory response,lessen cardiac dysfunction and protect neurons against injury in hippocampus CA1area after restoration of spontaneous circulation (ROSC). Methods Animal models of cardiac arrest were established in 24 New Zealand rabbits,and those animals were randomly (random number) divided into control group and UTI treated group after ROSC.Changes in the levels of plasma inflammatory cytokines TNF-α and IL-6 were assayed before cardiac arrest and 4,8,12 and 16 hours after ROSC.Cardiac function including FS,EF and E/A were observed with ultrasonography before cardiac arrest and 4,8,12 and 16hours after ROSC,and viable and apoptotic neurons in hippocampus CA1 area and infiltrations of MPO positive cells in myocardium,cerebrum,liver,kidney and intestine were counted 72 hours after ROSC.The t-test or Mann-Whitney rank sum test was used to verify the specified theoretical distribution functions of the biomarkers tested by Kolmogorov-Smirnov test,POST HOC test was used for the multiple comparisons,and Pearson correlation analysis was used to investigate the correlation between inflammatory cytokines and cardiac function. Results The levels of TNF-α and IL-6 in UTI group were lower than those in control group as those data got 4,8,12 and 16 hours after ROSC (P ＜0.05).EF and E/A in UTI treated group were higher than those in the control group as those data got 4,8,12 hours after ROSC.FS values obtained 4 h and 8 hours after ROSC were higher in UTI group than those in control group ( P ＜ 0.05 ).The Pearson correlation analysis showed that the levels of TNF-α and IL-6 significantly correlated with EF after ROSC.The number of viable neurons in CA1 area of control group was ( 13.22 ± 0.97) which was lower than that in UTI group ( 16.89 ± 1.45 ) ( P =0.003 ),while the number of apoptotic neurons in hippocampus CA1 area was higher in control group than that in UTI group (15.67 ± 1.37) vs.(13.67 ± 1.03 ) (P =0.019).The numbers of MPO positive cells were significantly lower in liver,kidney and intestine in group UTI than those in control group. Conclusions UTI could inhibit the infiltration of MPO positive cells in liver,kidney and intestine,decreasing the levels of TNF-α and IL-6 in plasma,in turn lessening cardiac dysfunction and protecting neurons from injury in hippocampus CA1 area after ROSC of New Zealand rabbits.
ABSTRACT
Objective To explore the effects of pyrrolidine dithiocarbamate (PDTC) on the acute lung injury and the activation of Nrf2 pathway after Paraquat (PQ) induced lung injury.Methods Fortyeight adult male SD rats with lung injury induced by PQ were randomly (random number) divided into control group and PDTC group.Three animals were sacrificed at every 1-week interval,7d,14d and 21 days after PQ intoxication,and the lungs of rats were removed for acute lung injury score after HE staining,and for lung fibrosis assessment after Masson staining,and the levels of reduced glutathione (GSH) and malondialdehyde (MDA) in the lung tissue homogenate were assayed and the phosphorylation of Nrf2 (nuclear-E2-related factor 2) was detected by Weston blot.The mean values of detected variables between two groups were compared by t test,and survival curve was tested by Wilcoxon (Gehan) test.Results The intoxication symptoms of rats were obvious,and 4 rats in control group and 9 rats in PDTC group survived until 21days.The survival time of animals in PDTC group was longer than that in control group (Wilcoxon (Gehan) =10.17023,P =0.001).The levels of MDA in control group were higher than those in PDTC group,while the levels of GSH in control group were lower than those in PDTC group.The levels of phosphorylation of Nrf2 in PDTC group were higher than those in control group at 1-week intervals,1-week:(0.32±0.04) vs.(0.23±0.05),P=0.003; 2-week:(0.62±0.06) vs.(0.33±0.03),P＜0.001; 3-week:(0.61 ±0.04) vs.(0.33±0.05),P＜0.001.The acute lung injury (ALI) scores in PDTC group were lower than those in control group,1-week:(5 ± 0.95) vs.(8 ± 1.23),P =0.002 ; 2-week:(9±1.18) vs.(11±1.02),P=0.019; 3-week:(11±1.33) vs.(12±1.42),P=0.002.The percentages of lung fibrosis at 1-week intervals after PQ intoxication were (40.87 ± 7.25) ％,(43.38 ±5.71)％ and (45.91 ± 3.97)％ in control group,and they were higher than those in PDTC group (32.92±2.34)％,(33.45 ±3.04)％ and (35.27 ±3.81)％ in PDTC group,P=0.017,0.001 and 0.001 respectively.Conclusions Attenuation of acute lung injury and lung fibrosis,and prolongation of survival time of SD rats by PDTC were associated with activation of Nrf2 pathway.