Objective To investigate the optimal injury time point of cardiac arrest (CA) induced electrically, and establish a reproducible prolonged CA and cardiopulmonary resuscitation (CPR) model in pigs. Methods Forty healthy domestic male pigs were randomly divided into four groups, which were ventricular fibrillation (VF) 8, 10, 11, and 12 minutes groups, each group for 10 animals. In these groups, VF was induced by alternating current delivered to right ventricular endocardium and untreated for 8, 10, 11, and 12 minutes, respectively, followed by 6 minutes of CPR procedure. The resuscitation and survival outcomes were recorded. Hemodynamic parameters and arterial blood gases of animals after successful resuscitation were measured and recorded for 6 hours. Those successful resuscitation animals were regularly evaluated for the neurological deficit score (NDS) and survival outcomes every 24 hours till 96 hours after resuscitation. Results The shortest duration of CPR (minute: 6.9±1.3) and the highest successful ratio of the first defibrillation (7/10) were observed in group VF 8 minutes, and the ratio of successful resuscitation was 100%. The best coronary perfusion pressure (CPP) during the CPR, less neurological impairment, longer survival time, more stable hemodynamics, and shorter time for arterial pH and lactate level restoring to the original state after CPR were also observed in group VF 8 minutes, and no severe damage was found in those animals. The longest duration of CPR (minute:10.3±2.9) and the lowest successful ratio of the first defibrillation (1/10) were observed in group VF 12 minutes, and only 4 animals achieved restoration of spontaneous circulation (ROSC), and no animal survived to CPR 96 hours. The worst CPP during CPR and the highest NDS after resuscitation were also found in VF 12 minutes animals compared to those animals in the other groups. The injuries caused by ischemia and hypoxia in groups VF 10 minutes and VF 11 minutes were in between those of the groups VF 8 minutes and VF 12 minutes, and the duration of CPR were (7.0±2.1) minutes and (8.2±2.6) minutes. There were 9 and 7 animals achieved ROSC in groups VF 10 minutes and VF 11 minutes correspondingly, and 6 and 4 animals survived to 96 hours respectively. Obviously unstable hemodynamics was observed during the period of CPR 2 hours in the two groups. At CPR 1 hour, the heart rates (HR, beats/min) in groups VF 10 minutes and VF 11 minutes increased to 172 (155, 201) and 168 (136, 196) respectively, and the mean arterial pressures (MAP, mmHg, 1 mmHg = 0.133 kPa) declined to 97 (92, 100) and 81 (77, 100), the cardiac output (CO, L/min) decreased to 5.0 (4.0, 5.8), 3.7 (3.0, 5.4) correspondingly. Distinct injuries were found in the two groups [CPR 24-96 hours NDS in groups VF 10 minutes and VF 11 minutes: 180 (110, 255)-20 (0, 400) and 275 (223, 350)-240 (110, 400)], and the arterial pH of the two group decreased to 7.26±0.09 and 7.23±0.09 respectively, and the level of lactate (mmol/L) increased to 9.17±1.48 and 12.80±2.71 correspondingly at CPR 0.5 hour. Significantly lower pH was observed in group VF 11 minutes compared to group VF 8 minutes at CPR 0.5 hour (7.23±0.09 vs. 7.33±0.04, P < 0.05). The highest level of lactate (mmol/L) was also found at the same time point in group VF 11 minutes, which recovered to normal slowly, and was still significantly higher than groups VF 8, 10, 12 minutes (7.58±3.99 vs. 2.55±1.53, 2.13±2.00, 3.40±2.30, all P < 0.05) at CPR 4 hours. Conclusions The longer duration of CA was, the more severe damage would be, the longer CPR time would be required, and the harder of the animals to achieve ROSC. In this prolonged CA and CPR porcine model, 10-11 minutes for untreated VF, was an optimal time point with appropriate successful rate of resuscitation, survival outcomes, and post-resuscitation injuries. Therefore, we recommended 10-11 minutes might be the rational length of no-flow time in this model.

Objective To study the effect of Yiqi Jiedu compound on autoimmune regulator(Aire)and transcription factor retinoic acid-related orphan receptor(ROR-γt)/Effects of forkhead wing helix transcription factor 3(Foxp3)-related inflammatory factor expression.Methods The EAMG model was replicated by immunization-induced injection of mouse-derived AchR-α subunit 97-116 peptide,including normal group,model group,positive control group(prednisone),and Yiqi Jiedu compound high-dose,medium-dose and low-dose groups.Group of 10,the behaviors and Lennon scores of the rats in each group were observed.ELISA method was used to detect serum transforming growth factor-β1(TGF-β1)and interleukin-6(IL-6)levels.Western blot was used to detect the thymus tissue of EAMG rats.The expression of Aire and Foxp3,and the expression of RORγt in the thymus tissue of rats in each group was detected by RT-PCR.Results Compared with before treatment,the Lennon grading score of each drug group was significantly decreased;compared with the prednisone group,there was no significant difference in the Lennon score of each dose group of Yiqi Jiedu compound(P>0.05).RT-PCR detection results showed that compared with the normal group,the relative expression of RORγt mRNA in the thymus of the model group was significantly increased(P<0.01).Western blot detection results showed that the expressions of Aire and Foxp3 in the model group were significantly lower than those in the normal group(P<0.05);The levels of Aire and Foxp3 proteins in thymus of the rats in each drug group were significantly higher than those of model group(P<0.01).ELISA results showed that serum IL-6 content in model group was significantly higher than that in normal group(P<0.01),while TGF-β1 content was significantly lower than that in normal group(P<0.01).After intragastric administration,the content of IL-6 in all drug groups was significantly decreased,and the content of TGF-β1 was significantly increased(P<0.01).Conclusion Yiqi Jiedu compound can improve the symptoms of muscle weakness in EAMG rats,and its mechanism may be through up-regulating the expression of Aire protein,regulating immune inflammatory response,affecting the differentiation of Th17/Treg,thereby regulating the immune balance of ROR-γt/Foxp3.The role of autoimmune response may be one of the mechanisms of its treatment of MG.

OBJECTIVE：To study the effects of chrysophanol on the activa tion of microglia and the expression of inflammatory factors in cerebral ischemia model rats. METHODS ：SD rats were randomly divided into sham operation group ， model group and chrysophanol high ，medium，low dose groups [7.88，3.94，1.97 mg/（kg·d）]，with 20 rats in each group （the number was complemented in cases of death or unsuccessful modeling during modeling process ）. Except for sham operation group ， middle cerebral artery occlusion model was established in other groups by improved thread method. After 2 hours of ischemia ， sham operation group and model group were intraperitoneally injected with 1 mL normal saline ，and each administration group was intraperitoneally injected with 1 mL corresponding drug ，once a day ，for 7 consecutive days. After last medication ，the score of neurological impairment was recorded ；cerebral infarction of rats was observed by TTC staining ，and the percentage of cerebral infarction area was calculated. The expression of Iba- 1 positive cells in ischemic penumbra of rats was observed by immunofluorescence staining. The expression of Notch- 1，TNF-α and ICAM-1 in the ischemic penumbra of rats were detected by Western blotting assay. RESULTS ：In sham operation group ，there was no infarction area in the brain tissue ，and the Iba- 1 positive cells in the ischemic penumbra were few and branched. Compared with sham operation group ，the infarction area of cerebral tissue in rats was obvious in model group ； the 052）number of Iba- 1 positive cells in ischemic penumbra were 〔ZQ2017003〕） increased significantly ，and they were in amoeba or round shape；the neurological impairment score ，the percentage of cerebral infarction area ， relative expression of Notch- 1， TNF-α and ICAM-1 protein in ischemic penumbra were increased significantly （P＜0.05）. Compared with m odel rats ，the infarction area of cerebral tissue in each dose group of chrysophanol was reduced to different extent ；the number of Iba- 1 positive cells in ischemic penumbra was decreased ；neurological impairment score ，the percentage of cerebral infarction area ，relative expression of Notch- 1，TNF-α and ICAM-1 protein were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Chrysophanol has a certain protective effect on the brain tissue of cerebral ischemia model rats ，and can relieve the nerve injury. Its mechanism may be associated with inhibiting the activation of microglia and expression of inflammatory factors mediated by Notch pathway.

Objective To observe the effects of Yiqi Jiedu Compound on TCM syndrome score and immune regulation of Th17/Treg cells in peripheral blood of patients with myasthenia gravis.Methods 63 patients with myasthenia gravis were randomly divided into the control group(n=31)and the treatment group(n=32).The control group was given the basic treatment of pyridostigmine bromide and glucocorticoid and so on,and the treatment group was given Yiqi Jiedu Compound based on the control group.The two groups were treated for 12 weeks.Clinical efficacy was observed,TCM syndrome scores were statistically analyzed,and the expression levels were detected,which Th17/Treg cells,Foxp3/RORγ t gene and IL-6 and IL-17 cytokines in peripheral blood of patients in both groups before and after treatment.Results After 12 weeks of treatment,compared with the control group,the TCM syndrome score of patients in the treatment group was significantly decreased(P<0.05),and the expression of Foxp3 and Treg genes in peripheral blood was promoted(P<0.05),and the expression of Th17 and Treg cells and IL-6 and IL-17 cytokines in peripheral blood was decreased(P<0.05).Conclusion Yiqi Jiedu Compound may regulate the immune balance of Th17/Treg cells by affecting the differentiation of Th17 and Treg cells in peripheral blood and the expression levels of transcription factors RORγ t and Foxp3 genes and related cytokines,thus achieving the effect of myasthenia gravis treatment.