Objective:To evaluate the effect of Alda-1 on ferroptosis in cardiomyocytes after cardiac arrest and cardiopulmonary resuscitation in swine.Methods:Twenty-two healthy male white swine, weighing 35-43 kg, were divided into 3 groups using a random number table method: sham operation group (group S, n=6), cardiac arrest-cardiopulmonary resuscitation group (group CA-CPR, n=8) and Alda-1 group ( n=8). The animals only underwent the general preparation in group S, and the swine model of cardiac arrest and cardiopulmonary resuscitation was developed by 8 min of electrically induced cardiac arrest through the pacing catheter in the right ventricle followed by 8 min of cardiopulmonary resuscitation in CA-CPR and Alda-1 groups.Alda-1 0.88 mg/kg was intravenously injected at 5 min after resuscitation in group Alda-1, and the equal volume of vehicle was administered instead in the other two groups.Stroke volume (SV) and global ejection fraction (GEF) were measured using PiCCO before developing the model and at 1, 2 and 4 h after resuscitation (T 0-3). Venous blood samples were collected from the femoral vein to measure the concentrations of serum cardiac troponin (cTnI) by enzyme-linked immunosorbent assay at T 0-3, and at 24 h after resuscitation (T 4). The animals were then sacrificed, and myocardial tissues in the left ventricle were harvested to measure the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) (by Western blot), iron deposition (by Prussian blue staining), 4-hydroxy-2-nonenal (4-HNE) content (by enzyme-linked immunosorbent assay), and malondialdehyde (MDA) and glutathione (GSH) contents (by colorimetry). Results:Compared with group S, SV and GEF were significantly decreased at T 1-3, the serum concentrations of cTnI were increased at T 1-4, myocardial ACSL4 expression was up-regulated, GPX4 expression was down-regulated, iron deposition and contents of 4-HNE and MDA were increased, and the content of GSH was decreased in CA-CPR and Alda-1 groups ( P<0.05). Compared with group CA-CPR, SV and GEF were significantly increased at T 2-3, the serum concentrations of cTnI were decreased at T 3-4, myocardial ACSL4 expression was down-regulated, GPX4 expression was up-regulated, iron deposition and contents of 4-HNE and MDA were decreased, and the content of GSH was increased in group Alda-1 ( P<0.05). Conclusions:Alda-1 can alleviate myocardial injury after cardiac arrest and cardiopulmonary resuscitation in swine and further improve cardiac dysfunction, and the mechanism may be related to inhibition of cell ferroptosis.

To establish a model of inactivation adenosine A2A receptors in brain tissues of mice, we transplanted bone marrow cells (BMCs) from wild type (WT) C57BL/6 mice into A2A receptor knockout (A2A KO) C57BL/6 mice which were previously fractionated total body irradiation of 6.2 Gyx2. Six weeks later, we identified and evaluated the model. The results showed that the sexual chromagene pattern on white blood cells of recipient mice changed from female pattern to male pattern and there were 95.9% of A2AR+ cells in peripheral white blood cells of recipient mice, whereas there was no significant difference of A2AR mRNA level in brains between these recipient mice and A2AR KO mice. Furthermore, there was no significant difference of breathing frequency, brain water content and level of glutamate between the model mice and WT mice. These results indicated that we established successfully a mouse model of inactivation adenosine A2A receptors in brain tissues. This may provide a new and efficient strategy to study the effect of adenosine A2A receptors in disease and injuries of central nervous system.
Animals ; Bone Marrow Transplantation ; Brain ; metabolism ; Disease Models, Animal ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Receptors, Adenosine A2 ; deficiency ; genetics ; Whole-Body Irradiation

Objective To study the characteristics of the sciatic nerve injuries inflicted by highvelocity triangle fragments in swine.Methods According to the injury position,14 swines weighing (34.29±5.2)kg were divided into two groups.All swines were shot by 0.37 g triangle fragments at (773.1±12.4)m/s aimed at the midpoint of the lateral body surface projection of the right sciatic nerve in Group A(n=7)and at 2 cm aside from sciatic nerve in Group B(n=7).The entrance,exit and length of wound tract,distance between nerve and wound tract were detected,and the pathology of sciatic nerve and wound tract at 48 hours after injury were observed under light microscope and electron microscope.Results The course of the fragments was deviated to different extent.Six sciatic nerves were in primary wound tract,four of which were lacerated and ruptured at different degrees.The light microscope showed pathological changes including severe hemorrhage and edema,leukocytic infiltration,neurotmesis,axonotmesis and light myelin coloration;while electron microscope showed severe degeneration of myelin sheath and neuraxon distortion.Six nerves(one in Group A and five in Group B)were in the concussion zone,with the distance between sciatic nerve and primary wound tract for(2.07±0.45)cm.Hyporrhea and hyperemia under perilemma and perineurium,partial nerve fiber disrupt and axonotmesis were observed by light microscope and partly myelin sheath delamination by electron microscope.Two nerves (one in Group A and another in Group B)were in the contusion zone,with moderate pathological changes.Conclusions The track of high velocity triangle fragments is unstable in the tissues.High disrupt rate,severe and wide extent lesion,early Wallerian degeneration are the main characteristics of direct injuries of swine's sciatic nerves.Indirect injuries can induce pathologic changes too,for which the lesion severity is related to the distance between nerves and primary wound tract.

OBJECTIVE: To investigate whether the acetaldehyde dehydrogenase 2 specific activator, Alda-1, can alleviate brain injury after cardiopulmonary resuscitation (CPR) by inhibiting cell ferroptosis mediated by acyl-CoA synthetase long-chain family member 4/glutathione peroxidase 4 (ACSL4/GPx4) pathway in swine. METHODS: Twenty-two conventional healthy male white swine were divided into Sham group (n = 6), CPR model group (n = 8), and Alda-1 intervention group (CPR+Alda-1 group, n = 8) using a random number table. The swine model of CPR was reproduced by 8 minutes of cardiac arrest induced by ventricular fibrillation through electrical stimulation in the right ventricle followed by 8 minutes of CPR. The Sham group only experienced general preparation. A dose of 0.88 mg/kg of Alda-1 was intravenously injected at 5 minutes after resuscitation in the CPR+Alda-1 group. The same volume of saline was infused in the Sham and CPR model groups. Blood samples were collected from the femoral vein before modeling and 1, 2, 4, 24 hours after resuscitation, and the serum levels of neuron specific enolase (NSE) and S100 β protein were determined by enzyme-linked immunosorbent assay (ELISA). At 24 hours after resuscitation, the status of neurologic function was evaluated by neurological deficit score (NDS). Thereafter, the animals were sacrificed, and brain cortex was harvested to measure iron deposition by Prussian blue staining, malondialdehyde (MDA) and glutathione (GSH) contents by colorimetry, and ACSL4 and GPx4 protein expressions by Western blotting. RESULTS: Compared with the Sham group, the serum levels of NSE and S100β after resuscitation were gradually increased over time, and the NDS score was significantly increased, brain cortical iron deposition and MDA content were significantly increased, GSH content and GPx4 protein expression in brain cortical were significantly decreased, and ACSL4 protein expression was significantly increased at 24 hours after resuscitation in the CPR model and CPR+Alda-1 groups, which indicated that cell ferroptosis occurred in the brain cortex, and the ACSL4/GPx4 pathway participated in this process of cell ferroptosis. Compared with the CPR model group, the serum levels of NSE and S100 β starting 2 hours after resuscitation were significantly decreased in the CPR+Alda-1 group [NSE (μg/L): 24.1±2.4 vs. 28.2±2.1, S100 β (ng/L): 2 279±169 vs. 2 620±241, both P < 0.05]; at 24 hours after resuscitation, the NDS score and brain cortical iron deposition and MDA content were significantly decreased [NDS score: 120±44 vs. 207±68, iron deposition: (2.61±0.36)% vs. (6.31±1.66)%, MDA (μmol/g): 2.93±0.30 vs. 3.68±0.29, all P < 0.05], brain cortical GSH content and GPx4 expression in brain cortical was significantly increased [GSH (mg/g): 4.59±0.63 vs. 3.51±0.56, GPx4 protein (GPx4/GAPDH): 0.54±0.14 vs. 0.21±0.08, both P < 0.05], and ACSL4 protein expression was significantly decreased (ACSL4/GAPDH: 0.46±0.08 vs. 0.85±0.13, P < 0.05), which indicated that Alda-1 might alleviate brain cortical cell ferroptosis through regulating ACSL4/GPx4 pathway. CONCLUSIONS Alda-1 can reduce brain injury after CPR in swine, which may be related to the inhibition of ACSL4/GPx4 pathway mediated ferroptosis.
Male ; Animals ; Swine ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Ferroptosis ; Brain Injuries ; Glutathione ; Cardiopulmonary Resuscitation ; Ligases ; Iron

Objective To summarize the efficacy and adverse reactions of incremental corticotrophin (ACTH) therapy in the treatment of infantile spasms (IS),and to provide new clinical treatment options.Methods The clinical data of 40 children with IS who were hospitalized in the Department of Neurology,Shanghai Children's Medical Center,Shanghai Jiaotong University School of Medicine,treated with ACTH from January 2016 to January 2018 were collected and retrospectively analyzed.All the children were treated with intravenous infusion of ACTH with an initial dose 12.5 U/d for 3 days.If the spasms did not disappear,dosage of ACTH increased to 25.0 U/d for another 3 days.If the spasms could not yet be fully controlled,the dosage increased to 40.0 U/d,and the total course of treatment did not exceed 2 weeks.If the spasms disappeared at each dose stage or the course of treatment reached to 2 weeks,ACTH would be changed to Prednisone 2 mg/(kg · d) orally,which gradually decreased in 2 months.All children underwent electroencephalogram examination before and after treatment.Results Forty patients with IS were treated with ACTH increasing therapy.The disappearance rate of spasms was 40.0％ (16/40 cases) totally,with 7.5％ (3/40 cases) at the dosage phase of 12.5 U/d,16.2％ (6/37 cases) at the dosage stage of 25.0 U/d,and 22.6％ (7/31 cases) at the dosage of 40.0 U/d.The disappearance rate of hypsarrhythmia on electroencephalogram was 60.0％ (24/40 cases) generally,and 5.0％ (2/40 cases),10.8％ (4/37 cases),58.1％ (18/31 cases),respectively at above different dosage phases,while 37.5％ (15/40 cases) of the children had mild adverse reactions,mostly respiratory infections.Conclusions The short-term efficacy of the ACTH incremental therapy in the treatment of IS is positive,and the incidence of adverse reactions is low.

Traumatic brain injury（TBI）in combination with its sequelae is currently a public health problem that seriously endangers human health. Accumulating studies have deeply investigated the pathological processes of TBI to find possible therapeutic targets. In recent years，the relationship between TBI pathogenesis and gut-brain axis changes in aspects of microflora，metabolites，inflammatory factors and neurohumoral changes has drawn widespread attention，which not only provides a better understanding how TBI affects gut-brain axis，but also provides new therapeutic targets in improving cure rate and recovery level after TBI. However，due to the existence of complicated gut-brain axis self-regulation as well as the individual differences of TBI patients，the recognition and transformation for the latest experimental research are largely limited. In this review，the authors summarize how TBI event influences gut-brain axis balance，as well as how gut-brain axis regulates TBI pathogenesis mutually，so as to provide a better understanding of the interaction between gut-brain axis and TBI.

Objective:To investigate the association between the Doppler variables of the ophthalmic artery with the severity of preeclampsia(PE).Methods:Systematic literature was searched between January 1995 and March 2023 in PubMed,Web of Science,Embase,and the Cochrane Library.Studies comparing ophthalmic artery Doppler variables,including peak systolic velocity(PSV),end-diastolic velocity(EDV),resistive index(Rl),pulsa-tility index(PI),and peak ratio(PR,the ratio of the flow velocity of the second peak to that of the initial peak)in patients with PE,severe preeclampsia(sPE),and healthy pregnant women were included.The random-effects model was adopted as the method of pooled analysis,and the I2value was used to assess heterogeneity.The pooled standardized mean difference(SMD)with 95％confidence interval(CI)was used to estimate the associa-tion between ophthalmic artery Doppler variables and PE patient's characteristics.Results:Eight retrospective studies were eventually included in this Meta-analysis.Our pooled results suggested that compared with PE ca-ses,sPE patients had lower PI levels(SMD-0.56,95％CI-0.92～-0.20,P=0.000),higher EDV levels(SMD 0.47,95％CI 0.12～0.83,P=0.028)and higher PR levels(SMD0.96,95％CI 0.13～1.78,P=0.023).Howev-er,there was no significant difference between PE and sPE patients about the PSV and RI(P=0.361,P=0.626).Conclusions:This review demonstrates that ophthalmic artery Doppler variables(PI,EDV and PR)could be useful for predicting PE and PE development(especially in identifying sPE),which in turn may help the practitioner in the management of these complicated cases and in taking early necessary precautions.