【Abstract】 Objective To isolate and culture human amniotic fluid-derived mesenchymal stem cells (HAFMSCs),to investigate a better cryopreservation protocol of HAFMSCs and to observe the biocharacteristics and the multi-potential of HAFMSCs after cryopreservation for the further fundamental researches and cl inical appl ications. Methods HAFMSCswere isolated from the amniotic fluid of pregnant women during the second trimester by the improved two-step method.HAFMSCs were cryopreserved with different cryopreservation protocols (containing different contents of FBS and DMSO atcryoprotectant) in l iquid nitrogen for 12 weeks. The biocharacteristics of the HAFMSCs after cryopreservation were analyzed. The growth characteristics were observed by MTT method and the growth curves were drawn. The surface antigens of HAFMSCs were detected using flow cytometry, including CD29, CD34, CD44, CD45, CD73, and CD90. The adi pogenic and osteogenic differentiation abil ities of HAFMSCs were observed. The mRNA levels of Oct-4 and Nanog of the HAFMSCs were compared between before and after cryopreservations. Results At 12 weeks after cryopreservation, different protocols had different effects on the cell viabil ity; the better formula of cryoprotectant was 50% DMEM, 40% FBS, and 10% DMSO. After cryopreservation, the cells proliferated rapidly and the growth curves showed “S” shape, which was the same as the cells before cryopreservation. Phenotype showed that HAFMSCs were positive for the surface markers CD29, CD44, CD73, and CD90, and negative for CD34 and CD45. After 21 days of adi pogenic differentiation, the l ipid droplets were observed by oil red O staining. After 21 days of osteogenic differentiation, the calcium mineralizations were verified by von Kossa staining. There was no significant difference (P gt; 0.05) in the mRNA levels of Oct-4 and Nanog between before and after cryopreservations. Conclusion HAFMSCs have rapid proliferation and multi-potential in vitro. The cells have high viabil ities and no changes of the biocharacteristics and differentiation potential ities after cryopreservation for 12 weeks. Cryoprotectant containing 50% DMEM, 40% FBS, and 10% DMSO is a better cryopreservation protocol.
Objective To investigate the relationship between thrombocytopenia after the restoration of spontaneous circulation and short-term prognosis of patients with in-hospital cardiac arrest. Methods The demographic data, post-resuscitation vital signs, post-resuscitation laboratory tests, and the 28-day mortality rate of patients who experienced in-hospital cardiac arrest at the Emergency Department of West China Hospital, Sichuan University between January 1st, 2016 and December 31st, 2016 were retrospectively analyzed. Logistic regression was used to analyze the correlation between thrombocytopenia after the return of spontaneous circulation and the 28-day mortality rate in these cardiac arrest patients. Results Among the 285 patients included, compared with the normal platelet group (n=130), the thrombocytopenia group (n=155) showed statistically significant differences in red blood cell count, hematocrit, white blood cell count, prothrombin time, activated partial thromboplastin time, and international normalized ratio (P<0.05). The 28-day mortality rate was higher in the thrombocytopenia group than that in the normal platelet group (84.5% vs. 71.5%, P=0.008). Multiple logistic regression analysis indicated that thrombocytopenia [odds ratio =2.260, 95% confidence interval (1.153, 4.429), P=0.018] and cardiopulmonary resuscitation duration [odds ratio=1.117, 95% confidence interval (1.060, 1.177), P<0.001] were independent risk factors for 28-day mortality in patients with in-hospital cardiac arrest. Conclusion Thrombocytopenia after restoration of spontaneous circulation is associated with poor short-term prognosis in patients with in-hospital cardiac arrest.
ObjectiveTo determine the effects of different volume fluid resuscitation on intestinal injury and the permeability of intestine in hemorrhagic shock rats. MethodsSprague-Dawley male rats(n=72) were randomly equally divided into 4 groups after the model establishment of blood pressure-controlled hemorrhage, 45, 30, and 15 mL/(kg·h) of fluid resuscitation were performed in high dosage of resuscitation(HLR), moderate dosage of resuscitation(MLR), and low dosage of resuscitation(LLR) group respectively, but rats of Sham group didn't accept fluid resuscitation. After resuscitation, ten centimeters ileum was harvested for testing intestinal permeability. Then 6 rats of each group were sacrificed at 24, 48, and 72 hours after fluid resuscitation respectively. Over the specified time interval, blood was collected for testing levels of lactic acid and plasma tumor necrosis factor-α(TNF-α). The ileums of 3 resuscitation groups were obtained for testing the ratio of wet weight to dry weight and observing the histological changes. ResultsAfter resuscitation, the intestinal permeability was higher in HLR group(P<0.05). At 3-8 hours after resuscitation, rats of Sham group were all died, and the other rats of 3 groups were all alive. The level of plasma lactic acid was lower in LLR group than those of other 2 groups at 24 hours(P<0.05). The levels of TNF-α were higher in HLR group than those of other 2 groups at 24, 48, and 72 hours(P<0.05), and at 48 hours, level of TNF-α in LLR group was lower than MLR group(P<0.05). At 24 hours after resuscitation, ratio of intestinal wet weight to dry weight in LLR group was the lowest, and HLR group was the highest(P<0.05). According to the histopathology, intestinal injuries of the 3 groups were tend to be remission with the time, and at 48 and 72 hours after resuscitation, intestinal villus of LLR group appeared to be normal. ConclusionLimited fluid resuscitation of 15 mL/(kg·h) could not only decrease the levels of lactic acid and TNF-α, but also moderate the intestinal permeability and the intestinal injury in early stage after shock and surgery.
Brain injury after cardiopulmonary resuscitation is closely related to the survival rate and prognosis of neurological function of cardiac arrest (CA) patients. Recently, the American Academy of Neurology (AAN) published a practice guideline which had updated the evaluation of different treatments for reducing brain injury following cardiopulmonary resuscitation. In order to master and transmit AAN 2017 practice guideline on reducing brain injury following cardiopulmonary resuscitation, this paper interprets the new AAN clinical practice guideline to assist Chinese clinicians for better studying the guideline.
ObjectiveTo analyze the long-term effect on cardiopulmonary resuscitation skill between video-led and scene simulation training and traditional instructor-led courses in medical student with eight-year program.MethodsNinety-nine medical students with eight-year program who studied in Peking Union Medical College were trained in cardiopulmonary resuscitation skill from January to February 2018. They were randomly divided into two groups, 53 students participated in basic life support course training, which belonged to video-led and scene simulation training as the trial group, and 46 students were trained by traditional instructor-led courses as the control group. In January 2019, the above 99 students were re-evaluated for cardiopulmonary resuscitation, and the outcome of cardiopulmonary resuscitation skill test in total scores and sub-items scores between two groups were compared. The data were analyzed using t test and Wilcoxon rank sum test.ResultsThe total average scores of the trial group (8.02±1.11) was higher than that of the control group (6.85±1.50) (P<0.05). The sub-items scores of the trial group in the three aspects of on-site assessment, chest compressions and simple respirators (1.64±0.37, 3.38±0.46, 1.52±0.58) were higher than those of the control group (1.33±0.45, 2.80±0.76, 1.19±0.58) (P<0.05). In terms of opening airway, there was no significant difference in scores between the two groups (1.02±0.47 vs. 1.10±0.45, P>0.05). The excellent rate of the trial group (60.3%) was significantly higher than that of the control group (30.4%) (P<0.05), and the unqualified rate (5.6%) was significantly lower than that of the control group (21.7%) (P<0.05).ConclusionsThe video-led and scene simulation training has a better effect on cardiopulmonary resuscitation skills acquisition and long-term maintenance than traditional instructor-led courses for medical student with eight-year program.
Objective To evaluate the effects and the clinical significances of liquid resuscitation on blood gas analysis， acid-base balance， electrolytes， acute physiology and chronic health evaluationsⅡ（APACHEⅡ） score of patients with septic shock， and then to analyze the relations between serum chlorine （Cl－） level and APACHEⅡscore and the volume of liquid resuscitation. Methods According to the target of resuscitation （centre venous pressure 8-12mm Hg and mean arterial pressure≥65mm Hg）， 21 patients with septic shock received enough fluid for resuscitation during 24h . The results of blood gas analysis， acid-base balance， electrolytes， and APACHE Ⅱ score were compared between pre-resuscitation and 24h post-resuscitation by self-controlled prospective study. The relationships of the level of serum Cl－ and APACHEⅡ score with the volume of liquid used in resuscitation were analyzed . Results The mean resus-citation duration was （18.09±4.57） h， and the volume of liquid during 24 h resuscitation was 5 320-11 028mL with mean volume of （7 775±1 735） mL in 21 patients with septic shock. Serum sodium （Na＋， mmol/L） and Cl－（mmol/L）levels of post-resuscitation were significant higher than those of pre-resuscitation （Na＋：138.71±5.67 versus 135.62±7.23， P=0.024；Cl－：109.10±4.90 versus 101.67±8.59， P=0.000）. Compared with the levels of pre-resuscitation， the blood pH value， hematocrit （Hct，%）， anion gap （AG， mmol/L）， lactic acid （mmol/L）， and APACHE Ⅱscore significantly decreased （pH：7.31±0.05 versus 7.37±0.06， P=0.000；Hct：28.48±2.56 versus 32.76±9.19， P=0.049；AG：8.33±3.45 versus 14.17±8.83， P=0.004；lactic acid：1.66±0.89 versus 2.96±1.23， P=0.001；APACHEⅡ：10.90±3.73 versus 17.24±4.06， P=0.000） after 24h resuscitation. The correlation analysis showed that the level of serum Cl- was positively correlated with the volume of liquid used in resuscitation （r=0.717，P＜0.01）. However， there was no correlation between APACHEⅡscore and the volume of liquid used in resuscitation （P＞0.05）. Conclusions The target of liquid resuscitation in patients with septic shock should be cautiously determined， including control of the volume of crystal liquid for resuscitation， in order to avoid acid-base imbalance or hyperchloraemia. At the same time， the change in internal environment should be monitored. An optimistic fluid resuscitation to decrease APACHE Ⅱ score in patients with septic shock is unrelated to the volume of liquid resuscitation.
Since the outbreak of the coronavirus disease 2019, the incidence and mortality of cardiac arrest have increased significantly worldwide, and the management of cardiac arrest is facing new challenges. The European Resuscitation Council issued the 2021 European Resuscitation Council Guidelines in March 2021 to update the important parts of cardiopulmonary resuscitation and added recommendations for the management of cardiopulmonary resuscitation during the coronavirus disease 2019 epidemic. This article will compare the difference between this guideline and the 2020 American Heart Association Guidelines for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care and integrate some key points, review literature and then summarize the latest research progress in cardiopulmonary resuscitation since the outbreak of the coronavirus disease 2019 epidemic. The content mainly involves cardiopulmonary resuscitation during the coronavirus disease 2019 epidemic, early prevention, early recognition, application of new technologies, airway management, extracorporeal cardiopulmonary resuscitation and post-resuscitation treatment.
Currently, cardiac arrest has become a major public health problem, which has a high incidence rate and a high mortality rate in humans. With the continuous advancement of cardiopulmonary resuscitation techniques, the overall prognosis of cardiac arrest victims is gradually improved. However, cardiac arrest events under special circumstances are still serious threats to human health. This article reviews the progress of epidemiology, pathogenesis, treatment characteristics, and key points of cardiopulmonary resuscitation in those special cardiac arrest events associated with trauma, poisoning, drowning and pregnancy.
ObjectiveTo investigate the effects of esophageal cooling (EC) on lung injury and systemic inflammatory response after cardiopulmonary resuscitation in swine.MethodsThirty-two domestic male white pigs were randomly divided into sham group (S group, n=5), normothermia group (NT group, n=9), surface cooling group (SC group, n=9), and EC group (n=9). The animals in the S group only experienced the animal preparation. The animal model was established by 8 min of ventricular fibrillation and then 5 min of cardiopulmonary resuscitation in the other three groups. A normal temperature of (38.0±0.5)℃ was maintained by surface blanket throughout the experiment in the S and NT groups. At 5 min after resuscitation, therapeutic hypothermia was implemented via surface blanket or EC catheter to reach a target temperature of 33℃, and then maintained until 24 h post resuscitation, and followed by a rewarming rate of 1℃/h for 5 h in the SC and EC groups. At 1, 6, 12, 24 and 30 h after resuscitation, the values of extra-vascular lung water index (ELWI) and pulmonary vascular permeability index (PVPI) were measured, and meanwhile arterial blood samples were collected to measure the values of oxygenation index (OI) and venous blood samples were collected to measure the serum levels of tumor necrosis factor-α (TNF-α) and inerleukin-6 (IL-6). At 30 h after resuscitation, the animals were euthanized, and then the lung tissue contents of TNF-α, IL-6 and malondialdehyde, and the activities of superoxide dismutase (SOD) were detected.ResultsAfter resuscitation, the induction of hypothermia was significantly faster in the EC group than that in the SC group (2.8 vs. 1.5℃/h, P<0.05), and then its maintenance and rewarming were equally achieved in the two groups. The values of ELWI and PVPI significantly decreased and the values of OI significantly increased from 6 h after resuscitation in the EC group and from 12 h after resuscitation in the SC group compared with the NT group (all P<0.05). Additionally, the values of ELWI and PVPI were significantly lower and the values of OI were significantly higher from 12 h after resuscitation in the EC group than those in the SC group [ELWI: (13.4±3.1) vs. (16.8±2.7) mL/kg at 12 h, (12.4±3.0) vs. (16.0±3.6) mL/kg at 24 h, (11.1±2.4) vs. (13.9±1.9) mL/kg at 30 h; PVPI: 3.7±0.9 vs. 5.0±1.1 at 12 h, 3.4±0.8 vs. 4.6±1.0 at 24 h, 3.1±0.7 vs. 4.2±0.7 at 30 h; OI: (470±41) vs. (417±42) mm Hg (1 mm Hg=0.133 kPa) at 12 h, (462±39) vs. (407±36) mm Hg at 24 h, (438±60) vs. (380±33) mm Hg at 30 h; all P<0.05]. The serum levels of TNF-α and IL-6 significantly decreased from 6 h after resuscitation in the SC and EC groups compared with the NT group (all P<0.05). Additionally, the serum levels of IL-6 from 6 h after resuscitation and the serum levels of TNF-α from 12 h after resuscitation were significantly lower in the EC group than those in the SC group [IL-6: (299±23) vs. (329±30) pg/mL at 6 h, (336±35) vs. (375±30) pg/mL at 12 h, (297±29) vs. (339±36) pg/mL at 24 h, (255±20) vs. (297±33) pg/mL at 30 h; TNF-α: (519±46) vs. (572±49) pg/mL at 12 h, (477±77) vs. (570±64) pg/mL at 24 h, (436±49) vs. (509±51) pg/mL at 30 h; all P<0.05]. The contents of TNF-α, IL-6, and malondialdehyde significantly decreased and the activities of SOD significantly increased in the SC and EC groups compared with the NT group (all P<0.05). Additionally, lung inflammation and oxidative stress were further significantly alleviated in the EC group compared with the SC group [TNF-α: (557±155) vs. (782±154) pg/mg prot; IL-6: (616±134) vs. (868±143) pg/mg prot; malondialdehyde: (4.95±1.53) vs. (7.53±1.77) nmol/mg prot; SOD: (3.18±0.74) vs. (2.14±1.00) U/mg prot; all P<0.05].ConclusionTherapeutic hypothermia could be rapidly induced by EC after resuscitation, and further significantly alleviated post-resuscitation lung injury and systemic inflammatory response compared with conventional surface cooling.
Cardiopulmonary resuscitation (CPR) is a very important treatment after cardiac arrest. The optimal treatment strategy of CPR is uncertain. With the accumulation of clinical medical evidence, the CPR treatment recommendations have been changed. This article will review the current hot issues and progress, including the pathophysiological mechanisms of CPR, how to achieve high-quality chest compression, how to achieve CPR quality monitoring, how to achieve optimal CPR for different individuals and how to use antiarrhythmic drugs.