OBJECTIVE: To investigate the mechanism of human embryonic stem cell-derived mesenchymal stem cells (hESC-MSC) in alleviating brain injury after resuscitation in swine with cardiac arrest (CA). METHODS: Twenty-nine healthy male large white swine were randomly divided into Sham group (n = 9), cardiopulmonary resuscitation (CPR) group (n = 10) and hESC-MSC group (n = 10). The Sham group only completed animal preparation. In CPR group and hESC-MSC group, the swine model of CA-CPR was established by inducing ventricular fibrillation for 10 minutes with electrical stimulation and CPR for 6 minutes. At 5 minutes after successful resuscitation, hESC-MSC 2.5×10⁶/kg was injected via intravenous micropump within 1 hour in hESC-MSC group. Venous blood samples were collected before resuscitation and at 4, 8, 24, 48 and 72 hours of resuscitation. The levels of neuron specific enolase (NSE) and S100B protein (S100B) were detected by enzyme linked immunosorbent assay (ELISA). At 24, 48 and 72 hours of resuscitation, neurological deficit score (NDS) and cerebral performance category (CPC) were used to evaluate the neurological function of the animals. Three animals from each group were randomly selected and euthanized at 24, 48, and 72 hours of resuscitation, and the hippocampus tissues were quickly obtained. Immunofluorescence staining was used to detect the distribution of hESC-MSC in hippocampus. Immunohistochemical staining was used to detect the activation of astrocytes and microglia and the survival of neurons in the hippocampus. The degree of apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). RESULTS: The serum NSE and S100B levels of brain injury markers in CPR group and hESC-MSC group were significantly higher than those in Sham group at 24 hours of resuscitation, and then gradually increased. The levels of NSE and S100B in serum at each time of resuscitation in hESC-MSC group were significantly lower than those in CPR group [NSE (μg/L): 20.69±3.62 vs. 28.95±3.48 at 4 hours, 27.04±5.56 vs. 48.59±9.22 at 72 hours; S100B (μg/L): 2.29±0.39 vs. 3.60±0.73 at 4 hours, 2.38±0.15 vs. 3.92±0.50 at 72 hours, all P < 0.05]. In terms of neurological function, compared with the Sham group, the NDS score and CPC score in the CPR group and hESC-MSC group increased significantly at 24 hours of resuscitation, and then gradually decreased. The NDS and CPC scores of hESC-MSC group were significantly lower than those of CPR group at 24 hours of resuscitation (NDS: 111.67±20.21 vs. 170.00±21.79, CPC: 2.33±0.29 vs. 3.00±0.00, both P < 0.05). The expression of hESC-MSC positive markers CD73, CD90 and CD105 in the hippocampus of hESC-MSC group at 24, 48 and 72 hours of resuscitation was observed under fluorescence microscope, indicating that hESC-MSC could homing to the damaged hippocampus. In addition, compared with Sham group, the proportion of astrocytes, microglia and apoptotic index in hippocampus of CPR group were significantly increased, and the proportion of neurons was significantly decreased at 24, 48 and 72 hours of resuscitation. Compared with CPR group, the proportion of astrocytes, microglia and apoptotic index in hippocampus of hESC-MSC group decreased and the proportion of neurons increased significantly at 24 hours of resuscitation [proportion of astrocytes: (14.33±1.00)% vs. (30.78±2.69)%, proportion of microglia: (12.00±0.88)% vs. (27.89±5.68)%, apoptotic index: (12.89±3.86)% vs. (52.33±7.77)%, proportion of neurons: (39.44±3.72)% vs. (28.33±1.53)%, all P < 0.05]. CONCLUSIONS Application of hESC-MSC at the early stage of resuscitation can reduce the brain injury and neurological dysfunction after resuscitation in swine with CA. The mechanism may be related to the inhibition of immune cell activation, reduction of cell apoptosis and promotion of neuronal survival.
Animals ; Heart Arrest/therapy* ; Cardiopulmonary Resuscitation ; Swine ; Humans ; Male ; Human Embryonic Stem Cells/cytology* ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stem Cells/cytology* ; Phosphopyruvate Hydratase/blood* ; Brain Injuries/therapy* ; S100 Calcium Binding Protein beta Subunit ; Apoptosis ; Disease Models, Animal

Objective To discuss the safety and effectiveness of ProGlide vascular stitching device in catheter-directed thrombolysis for patients with thrombosis of lower limb arteries.Methods The patients with arterial thrombosis of lower extremity,who received catheter-directed thrombolysis at the Department of Vascular Surgery of Weifang People's Hospital of China from January 2020 to December 2023,were collected for this study.In performing the vascular interventional surgery,a 6-Fr femoral artery sheath was used to accomplish the catheter-directed thrombolysis in all patients.When the catheter-directed thrombolysis was ended,ProGlide vascular stitching device was used to suture the femoral artery puncture point,or the femoral artery puncture point was oppressed with a finger.The hemostatic success rate and the incidence of relevant complications were compared between the two groups.Results A total of 156 patients who received catheter-directed thrombolysis of lower extremity were enrolled in this study.Of the 156 patients,ProGlide vascular stitching device was used to suture the femoral artery puncture point in 80(ProGlide device group)and finger-oppression was adopted in 76(finger-oppression group).Compared with finger-oppression group,in ProGlide device group the hemostatic success rate was obviously higher(96.25％vs.80.3％,P=0.002).Nine patients in finger-oppression group(11.8％)and 2 patients in ProGlide device group(2.5％)developed puncture site hematoma or pseudoaneurysm(P=0.023).Five patients in finger-oppression group(6.6％)had to receive surgical operation to repair the femoral artery,and one patient in ProGlide device group(1.25％)developed ProGlide device-related femoral artery occlusion.Adverse events occurred in 15 patients(19.7％)of finger-oppression group and 3 patients(3.75％)of ProGlide device group(P=0.002).Conclusion ProGlide vascular stitching device can be safely used in patients with arterial thrombosis of lower extremity receiving catheter-directed thrombolysis.This device carries high success rate for suturing the femoral artery puncture point,thus,the incidence of hematoma at the puncture site can be reduced.

Objective:To investigate the efficacy of a domestically developed small esophageal cooling device in implementing targeted temperature management (TTM) after resuscitation and its impact on organ injury using a porcine model of cardiac arrest and resuscitation.Methods:Thirty healthy male domestic white pigs were randomly divided into four groups using a random number table: sham (S group, n=6), normothermia (NT group, n=8), surface cooling (SC group, n=8), and esophageal cooling (EC group, n=8). The S group underwent only surgical preparation, while the other groups were subjected to 12 minutes of ventricular fibrillation followed by 6 minutes of cardiopulmonary resuscitation to establish cardiac arrest. The S and NT groups maintained a core temperature of (37.5±0.5)°C using a surface blanket. In the SC and EC groups, therapeutic hypothermia was induced post-resuscitation via surface blanket or esophageal cooling catheter to achieve a target temperature of 34°C, maintained the target temperature (34±0.5)°C for 6 hours, followed by controlled rewarming at 0.5°C/h to 37°C. Core temperature was continuously monitored for 12 hours post-resuscitation. Hemodynamic parameters, including stroke volume (SV), global ejection fraction (GEF), extravascular lung water index (ELWI), and pulmonary vascular permeability index (PVPI), were assessed using pulse indicator continuous cardiac output (PiCCO) monitoring. Serum levels of cardiac troponin I (cTnI), neuron-specific enolase (NSE), creatinine (Cr), and intestinal fatty acid-binding protein (IFABP) were measured via ELISA at 2, 6, 12, and 24 hours post-resuscitation. Neurological outcomes were evaluated at 24 hours using the neurological deficit score (NDS) and cerebral performance category (CPC). Continuous variables were analyzed using one-way ANOVA. Results:During TTM, the EC group exhibited a faster cooling rate [(1.52±0.18)°C/h vs. (0.94±0.32)°C/h, P<0.05] and shorter time to target temperature [(2.32±0.43) h vs. (3.78±0.82) h, P<0.05] compared to the SC group, with comparable maintenance and rewarming ( P>0.05). Compared to the S group, the NT, SC, and EC groups demonstrated significant post-resuscitation multi-organ injury, characterized by reduced SV and GEF, elevated ELWI and PVPI, and increased serum cTnI, NSE, Cr, IFABP, NDS, and CPC scores (all P<0.05). Relative to the NT group, the SC and EC groups showed improved SV (at 1 h post-resuscitation), GEF (at 1, 2, 4, and 6 h), ELWI (at 12 h), and reduced cTnI and NSE (at 6 h), Cr and IFABP (at 2 h), and NDS and CPC (at 24 h) (all P<0.05). Compared to the SC group, the EC group exhibited lower PVPI (at 12 h), reduced cTnI, Cr, and IFABP (at 2 h), decreased NSE (at 2, 12, and 24 h), and improved NDS (at 24 h) (all P<0.05). Conclusions:In a porcine model of cardiac arrest and resuscitation, the domestic esophageal cooling device facilitated rapid induction, stable maintenance, and controlled rewarming during TTM, outperforming traditional surface cooling. This approach demonstrated superior organ protection, warranting further investigation.

Objective:To explore the protective efficacy of sulforaphane (SFN) in alleviating intestinal mucosal injury after resuscitation in pigs and its possible mechanism.Methods:This experiment was performed in the laboratory animal center, Zhejiang university. Using a random number table, twenty-four domestic healthy male white pigs were randomly divided into the Sham group, cardiopulmonary resuscitation (CPR) group, and SFN group, in which the Sham group had 6 pigs, and the other two groups had 9 pigs, respectively. The experimental parameters of 10 min of cardiac arrest and 6 min of CPR were chosen to establish the porcine model of CPR in the CPR and SFN groups. At 5 min after resuscitation, a dose of 2 mg/kg of SFN was infused via the femoral vein within 10 min in the SFN group. At 1 h, 2 h, 4 h, and 24 h after resuscitation, vein samples were collected, and then the levels of intestinal fatty acid binding protein (IFABP) and diamine oxidase (DAO) in serum were measured by ELISA. Subsequently, 6 pigs were chosen to be euthanized in each group, and then tissue samples were harvested from distal ileum to measure the level of cell apoptosis by TUNEL, the activities of superoxide dismutase (SOD) and catalase (CAT) and the contents of glutathione (GSH) and malondialdehyde (MDA) by biochemical method, the contents of 4-hydroxy-2-nonenal (4-HNE) by ELISA, the fluorescence intensity of reactive oxygen species (ROS) by immunofluorescence staining, and the expression levels of zonula occluden-1 (ZO-1), occludin, nuclear factor E2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) by Western blot. Continuous variables were compared with one way analysis of variance among the three groups, and Bonferroni test was used for further pairwise comparison.Results:During the observation period after resuscitation, the serum levels of biomarkers of intestinal mucosal injury including IFABP and DAO were significantly higher in the CPR and SFN groups than in the Sham group (all P<0.05). However, the serum levels of IFABP at 2 h, 4 h, and 24 h after resuscitation and the serum levels of DAO at 1 h, 2 h, 4 h, and 24 h after resuscitation were significantly lower in the SFN group than in the CPR group (all P<0.05). At 24 h after resuscitation, apoptotic index was significantly increased, SOD and CAT activities and GSH contents were significantly decreased, MDA and 4-HNE contents and ROS production were significantly increased, ZO-1 and occludin expression were significantly down-regulated, and Nrf2 and HO-1 expression were significantly up-regulated in the CPR and SFN groups when compared with the Sham group (all P<0.05). However, apoptotic index was significantly decreased, SOD and CAT activities and GSH contents were significantly increased, MDA and 4-HNE contents and ROS production were significantly decreased, and ZO-1, occludin, Nrf2, and HO-1 expression were significantly up-regulated in the SFN group when compared to the CPR group (all P<0.05). Conclusion:SFN could effectively protect against intestinal mucosal injury after resuscitation in pigs, and its mechanism was possibly related to the inhibition of oxidative stress and cell apoptosis via the activation of Nrf2/HO-1 pathway.

Objective:To establish pig cardiac arrest resuscitation model, and explore the effect of automatic compression synchronous ventilation (ACSV) on cardiopulmonary resuscitation in pigs.Methods:Twelve male white pigs with body weight of (38±3) kg were divided into ACSV group and intermittent positive pressure ventilation (IPPV) group with 6 pigs in each group by random number table method. A porcine cardiac arrest and resuscitation model was prepared with ventricular fibrillation induced by alternating current release via right ventricular electrode for 6 min and compression for 8 min. Mechanical chest external compression depth 5 cm, frequency 100 times/min. The tidal volume of ACSV group was 3 mL/kg and the frequency was 100 times/min. In the IPPV group, the tidal volume was 7 mL/kg and the frequency was 10 times/min. Arterial blood was drawn before resuscitation and at 1, 4 and 7min during resuscitation for blood gas analysis. Coronary perfusion pressure (CPP), end-respiratory carbon dioxide (ETCO 2) and carotid blood flow (CBF) were monitored during resuscitation. Stroke volume (SV) and global ejection fraction (GEF) were recorded by pressure monitoring catheter before and 1, 2 and 4 h after resuscitation. Venous blood samples were collected at each time point and 24 h after resuscitation to detect cardiac troponin I (cTnI), neuron specific enolase (NSE), alamine aminotransferase (ALT), creatinine (Cr), and intestinal fatty acid binding protein (IFABP). Results:(1) During resuscitation, CPP, ETCO 2 and CBF in ACSV group were slightly higher than those in IPPV group, but the differences between groups were not statistically significant. (2) There was no significant difference in pH, PaCO 2, HCO 3- and lactic acid between the two groups during resuscitation. The PaO 2 in ACSV group was higher than that in IPPV group, and the difference was statistically significant at 4 and 7 min. (3) The success rate of resuscitation in both groups was 83.3%, and there was no significant difference in SV and GEF before and after resuscitation. (4) After resuscitation, cTnI, NSE, ALT, Cr, iFABP and other indexes in ACSV group were lower than those in IPPV group, and there were statistically significant differences in cTnI at 24 h after resuscitation, ALT at 2 h and 24 h after resuscitation, and IFABP at 4 h and 24 h after resuscitation (all P<0.05). Conclusions:This study preliminarily suggested that the novel ACSV could significantly improve the oxygen supply level during cardiopulmonary resuscitation in pigs, while keeping the compression efficiency unchanged, avoiding hyperventilation, and reducing multiple organ damage after resuscitation, which is worthy of further study.

Objective To investigate the safety and efficacy of ExoSeal vascular occluder in bridging therapy for patients with acute ischemic stroke(AIS).Methods The clinical data of 142 AIS patients,who received mechanical thrombectomy in the bridge-connected vascular lumen after full dose recombinant tissue plasminogen activator(rt-PA)thrombolysis at the Weifang Municipal People's Hospital of China between June 2018 and December 2020,were retrospectively analyzed.An 8-F femoral arterial sheath was used for the performance of endovascular mechanical thrombectomy in all patients.After the interventional procedure finished,the hemostasis of femoral artery puncture point was achieved by using manual compression(MC)method in 68 patients(MC group)and by using a 7-F ExoSeal vascular occluder method in 74 patients(ExoSeal group).The success rate of puncture point hemostasis and the incidence of puncture point-related complications were compared between the two methods.Results The success rate of surgery in the ExoSeal group was 94.6％,which was obviously higher than 83.8％in the MC group(P=0.037).During the period from the time of finishing surgery and the time of discharge,11 patients(16.2％)in the MC group and 4 patients(5.4％)in the ExoSeal group developed puncture point-related complications(P=0.030),and 6 patients(8.8％)in the MC group and 2 patients(2.7％)in the ExoSeal group developed deep vein thrombosis(P=0.109).No arteriovenous fistula,pseudoaneurysm,acute ischemia of the ipsilateral lower limb,puncture point-associated major bleeding,or other complications that required vascular surgery or interventional therapy occurred in all patients.Conclusion ExoSeal vascular closure device can be safely used in AIS patients who are receiving mechanical thrombectomy in the bridge-connected vascular lumen after thrombolysis.It carries high success rate for femoral artery puncture point hemostasis,besides,it can reduce the incidence of puncture point hematoma.

Objective:To establish the pig model of cardiac arrest and resuscitation, and then investigate the protective role of sivelestat (SV) on the heart after resuscitation and its relation with β-catenin signaling pathway.Methods:Twenty-five healthy male white pigs were purchased. The animals were randomly divided into the Sham group ( n=6), cardiopulmonary resuscitation group (CPR, n=10), and CPR+SV group ( n=9). The experimental animal model was established by 9 min of cardiac arrest induced by the method of ventricular fibrillation and then 6 min of CPR in the CPR and CPR+SV groups. At 5 min after successful resuscitation, a dose of 10 mg/kg of SV was infused in a duration of 1h via the femoral vein with a micro-infusion pump in the CPR+SV group. Myocardial function evaluated by the values of stroke volume (SV) and global ejection fraction (GEF) was measured by PiCCO at baseline, and at 0.5, 1, 2, 4 h after resuscitation. The serum concentrations of cardiac injury biomarkers including cardiac troponin I (cTnI) and creatine kinase isoenzymes (CK-MB) were measured by ELISA using blood samples drawn from the femoral vein at baseline, and at 1, 2, 4, and 24 h after resuscitation. The animals were euthanized at 24 h after resuscitation, and then cardiac tissue samples were harvested to measure the protein expression levels of β-catenin, Cyclin D1, c-Myc, cleaved caspase-9, and cleaved caspase-3 by Western blot and the degree of cell apoptosis by TUNEL. Results:Prior to cardiac arrest, myocardial function and cardiac injury biomarkers were maintained at the same levels, and no differences were observed among the three groups (all P> 0.05). After resuscitation, myocardial dysfunction and cardiac injury were observed in the CPR and CPR+SV groups, in which the values of SV and GEF were significantly decreased and meanwhile the serum concentrations of cTnI and CKMB were significantly increased when compared with the Sham group (all P< 0.05). However, myocardial dysfunction and cardiac injury were significantly milder in the CPR+SV group, in which the value of SV at 4h post-resuscitation and the values of GEF starting 1h post-resuscitation were significantly increased, and the serum concentrations of cTnI and CKMB were significantly decreased at 4 and 24 h post-resuscitation when compared to the CPR group (all P< 0.05). Tissue measurements indicated that the change of β-catenin signaling pathway and the occurrence of cell apoptosis were observed in the heart at 24 h post-resuscitation in the CPR and CPR+SV groups, which were indicated by significant increases in the protein expression levels of β-catenin, Cyclin D1, c-Myc, cleaved caspase-9, and cleaved caspase-3, and marked elevation in the index of cell apoptosis when compared with the Sham groups (all P< 0.05). However, the expression levels of proteins mentioned above were significantly decreased in the heart at 24 h post-resuscitation and the index of cell apoptosis was significantly reduced in the CPR+SV group when compared to the CPR group (all P< 0.05). Conclusion:SV has the protective role in alleviating post-resuscitation myocardial dysfunction and cardiac injury, in which the protective mechanism is possibly related to the alleviation of cell apoptosis through the inhibition of β-catenin signaling pathway activation.

Objective To compare and analyze the difference of cardiopulmonary reserve function of phlegm-dampness and peaceful constitutions. Methods A total of 102 cases were selected for the physi-cal examination in Air Service Department of Northern Theater Air Force Hospital,and divided into a phlegm-dampness constitution group of 50 and a peaceful constitution group of 52 according to their traditional Chinese medicine constitution. Both groups underwent the cardiopulmonary exercise testing (CPET),and the cardiopulmonary reserve function parameters such as the peak exercise load,peak metabolic equivalent,peak kilogram oxygen uptake,and peak oxygen pulse rate were compared be-tween the 2 groups. Results There was no significant difference between the 2 groups in age,as well as the history of smoking and drinking(P>0.05). Moreover,the average body mass index,neck circum-ference,heart rate reserve,and peak ventilation of the phlegm-dampness constitution group were signif-icantly higher than the peaceful constitution group,while the average peak exercise load,peak meta-bolic equivalent,peak heart rate,peak oxygen pulse,and peak kilogram oxygen uptake were signifi-cantly lower than the latter group(P<0.05). Conclusions Compared with the people of peaceful constitu-tion,those of phlegm-dampness have lower cardiopulmonary reserve function. Therefore,early physical intervention should be conducted to help them to get into a healthy lifestyle.

Objective To compare and analyze the difference of cardiopulmonary reserve function of phlegm-dampness and peaceful constitutions. Methods A total of 102 cases were selected for the physi-cal examination in Air Service Department of Northern Theater Air Force Hospital,and divided into a phlegm-dampness constitution group of 50 and a peaceful constitution group of 52 according to their traditional Chinese medicine constitution. Both groups underwent the cardiopulmonary exercise testing (CPET),and the cardiopulmonary reserve function parameters such as the peak exercise load,peak metabolic equivalent,peak kilogram oxygen uptake,and peak oxygen pulse rate were compared be-tween the 2 groups. Results There was no significant difference between the 2 groups in age,as well as the history of smoking and drinking(P>0.05). Moreover,the average body mass index,neck circum-ference,heart rate reserve,and peak ventilation of the phlegm-dampness constitution group were signif-icantly higher than the peaceful constitution group,while the average peak exercise load,peak meta-bolic equivalent,peak heart rate,peak oxygen pulse,and peak kilogram oxygen uptake were signifi-cantly lower than the latter group(P<0.05). Conclusions Compared with the people of peaceful constitu-tion,those of phlegm-dampness have lower cardiopulmonary reserve function. Therefore,early physical intervention should be conducted to help them to get into a healthy lifestyle.

Objective:To provide references for further assessment of fighter pilots′ physical status by comparing and analyzing the difference of cardiopulmonary reserve function between different performance fighter aircraft pilots.Methods:A total of 96 fighter pilots were selected in the annual physical examination of the Air Service Department of Northern Theater Air Force Hospital. They were divided into the high-performance fighter group and the ordinary fighter group according to the aircraft types. Cardiopulmonary exercise testing (CPET) was performed to compare the basic parameters and cardiopulmonary reserve function parameters such as peak equivalent, peak oxygen pulse and peak kilogram oxygen uptake in the CPET monitoring results of the 2 groups.Results:There was no significant difference in general information between the 2 groups (all P>0.05). There was no significant difference between the high-performance fighter pilots and the ordinary fighter pilots in the main exercise cardiopulmonary reserve function parameters such as peak exercise load, peak oxygen pulse and peak kilogram oxygen uptake (all P>0.05). Conclusions:The general performance of the ordinary fighter pilots is the same as that of the high-performance fighter pilots in terms of exercise cardiopulmonary reserve function, and the CPET results can also reflect the status of pilot′s physical training.