Objective:To explore the application effect of the forgetting curve combined with blended learning in extracorporeal membrane oxygenation (ECMO) nursing training for ICU nurses.Methods:A randomized controlled trial was conducted from June to November 2022. Sixty-three ICU nurses from the First Hospital of Quanzhou City were selected using cluster sampling and divided into an observation group ( n=30) and a control group ( n=33). The control group received traditional teaching methods for training, while the observation group received training using the forgetting curve combined with blended learning. Compared the ECMO theoretical scores and skill operation scores of two groups of nurses after 1 day, 1 month, and 3 months of training; observed the core competencies and training satisfaction of ECMO nurses before and after training in two groups of nurses. Results:In the control group, there were 7 males and 26 females with an average age of (27.88 ± 4.36) years old; in the observation group, there were 6 males and 24 females with an average age of (28.67 ± 4.24) years old.Prior to training, there were no statistically significant differences in theoretical scores, skill operation scores, and core competencies between the two groups (all P>0.05). After 1 day, 1 month, and 3 months of training, the observation group′s ECMO theoretical scores were (80.33 ± 3.11), (78.13 ± 3.70), (76.73 ± 3.30) points respectively, higher than those of the control group which were (75.18 ± 3.30), (73.88 ± 2.75), (70.48 ± 2.96) points, with statistically significant differences ( t=6.36, 5.21, 7.92, all P<0.01); the observation group′s ECMO skill operation scores were (84.10 ± 4.16), (82.73 ± 3.71), (81.50 ± 3.40) points respectively, higher than the control group′s (78.09 ± 4.30), (74.97 ± 4.17), (71.85 ± 4.03) points, with statistically significant differences ( t=5.63, 7.77, 10.22, all P<0.01). There were statistically significant differences between two groups of nurses in terms of ECMO theory score and skill operation score, as well as time and interactive effect after training ( Finteraction=11.16, 84.76, both P<0.05). After training, the observation group′s total score for ECMO nurse core competency was (280.23 ± 9.23) points, superior to the control group′s (245.39 ± 14.90) points, with a statistically significant difference ( t=11.26, P<0.01). The observation group′s satisfaction total score and the scores in various dimensions were (99.17 ± 10.79), (4.43 ± 0.50), (4.30 ± 0.53), (4.57 ± 0.68), (4.37 ± 0.67), (4.23 ± 0.57) points, all higher than those of the control group which were (84.30 ± 12.61), (3.67 ± 0.96), (3.48 ± 0.71), (3.67 ± 0.74), (3.73 ± 0.72), (3.82 ± 0.77) points, with statistically significant differences ( t values were 2.42 to 5.09, all P<0.05). Conclusions:The application of the forgetting curve combined with blended learning in ECMO nursing training for ICU nurses is scientifically feasible. It helps nurses master ECMO theory and skill operations, improve ECMO nurse core competencies, enhance training satisfaction, and provides a new method for ECMO nursing training.

Objective To explore the potential categories of self-regulatory fatigue among maintenance hemodialysis patients and their relationships with self-management. Methods A total of 502 maintenance hemodialysis patients in Lanzhou were surveyed using a general information questionnaire, a Self-Regulatory Fatigue Scale, and a Self-Management Scale for Hemodialysis Patients. Potential profile analysis was conducted on self-regulatory fatigue, and its relationship with self-management was explored. Results Based on their self-regulatory fatigue status, maintenance hemodialysis patients could be categorized into four potential groups: low self-regulatory fatigue group (5.0%), moderate self-regulatory fatigue group (49.2%), high self-regulatory fatigue group (13.9%), and high cognitive regulatory fatiguegroup (31.9%). Gender, education level, and monthly per capita family income were identified as influencing factors for the potential categories of self-regulatory fatigue. Significant differences were observed in the total self-management scores among patients belonging to different potential categories of self-regulatory fatigue (F=50.617, P < 0.001). Conclusion Maintenance hemodialysis patients can be classified into four potential categories of self-regulatory fatigue, with patients in the low self-regulatory fatigue group demonstrating higher levels of self-management. Reducing the level of self-regulatory fatigue can contribute to enhancing patients'self-management capabilities.

Objective:To evaluate the effect of esketamine on hippocampal neuronal necroptosis in aged rats with postoperative cognitive dysfunction.Methods:One hundred and twenty SPF-grade healthy male Sprague-Dawley rats, aged 22 months, weighing 550-600 g, were divided into 4 groups ( n=30 each) using a random number table method: control group (group C), postoperative cognitive dysfunction group (group P), postoperative cognitive dysfunction+ esketamine group (group PE), and esketamine group (group CE). Rats received exploratory laparotomy under sevoflurane anesthesia, and esketamine 10 mg/kg and the equal volume of 0.9% sodium chloride were intraperitoneally injected at the end of surgery once a day for 6 consecutive days in group P and group PE, respectively. Rats received no anesthesia and surgery, and esketamine 10 mg/kg and the equal volume of 0.9% sodium chloride were intraperitoneally injected at the end of surgery once a day for 6 consecutive days in group CE and group C, respectively. Morris water maze test was performed at 7th day after surgery. The escape latency, times of crossing the original platform and time spent in the original platform quadrant were recorded. The rats were sacrificed at the end of Morris water maze test, and the hippocampal tissues were collected for determination of the rate of necroptosis and cytosolic Ca 2+ concentrations (by flow cytometry) and expression of mixed lineage kinase domain-like protein (MLKL), phosphorylated MLKL (p-MLKL), receptor-interacting protein kinase-3 (RIPK3), phosphorylated RIPK3 (p-RIPK3), receptor-interacting protein kinase-1 (RIPK1) and phosphorylated RIPK1 (p-RIPK1) (by Western blot). Results:Compared with group C, the escape latency was significantly prolonged, the times of crossing the original platform were decreased, the time spent in the original platform quadrant was shortened, the necroptosis rate of hippocampal neurons and cytosolic Ca 2+ concentrations were increased, and the expression of MLKL, p-MLKL, RIPK3, p-RIPK3, RIPK1 and p-RIPK1 was up-regulated in group P and group PE ( P<0.05). Compared with group P, the escape latency was significantly shortened, the times of crossing the original platform were increased, the time spent in the original platform quadrant was prolonged, the necroptosis rate of hippocampal neurons and cytosolic Ca 2+ concentrations were decreased, and the expression of MLKL, p-MLKL, RIPK3, p-RIPK3, RIPK1 and p-RIPK1 was down-regulated in group PE ( P<0.05). Conclusions:The mechanism by which esketamine attenuates postoperative cognitive dysfunction may be related to inhibition of necroptosis in hippocampal neurons of aged rats.

Objective:To evaluate the effect of necrostatin-1 (Nec-1)pre-treatment on postoperative cognitive function in aged rats with chronic pain due to knee arthritis.Methods:One hundred and twenty healthy male Sprague-Dawley rats, aged 22 months, weighing 550-600 g, were divided into 4 groups ( n=30 each) using a random number table method: chronic pain due to knee arthritis group(group P), chronic pain due to knee arthritis + operation group (group PS), dimethyl sulfoxide (DMSO) + chronic pain due to knee arthritis + operation group (DMSO+ PS group), and necrostatin-1 + chronic pain due to knee arthritis + operation group (Nec-1+ PS group). The inflammation-induced knee arthritis model was developed by injecting monosodium iodoacetate (MIA) into the left joint cavity.The exploratory laparotomy under sevoflurane anesthesia was performed at 12 weeks after intra-articular MIA injection. In Nec-1+ PS group and DMSO+ PS group, necrosstatin-1 6.25 mg/kg and the equal dose of DMSO were intraperitoneally injected at 1 h before surgery, respectively. At 7 days after surgery, the Morris water maze test was used to evaluate the cognitive function, the activation of microglial cells in the dentate gyrus of hippocampus was observed by immunofluorescent staining, and the activation rate of microglia cells was calculated, the necrosis rate of neurons was determined by flow cytometry, the expression of receptor-interacting protein kinase 1 (RIPK1)and phosphorylated mixed lineage kinase domain-like protein (p-MLKL) was determined by Western blot, and the contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 were determined by enzyme-linked immunosorbent assay. Results:Compared with P group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, and the number of crossing the original platform was reduced, the activation rate of microglia cells in the hippocampal dentate gyrus and necrosis rate of hippocampal neurons were increased, the expression of RIPK1 and p-MLKL was up-regulated, and the contents of pro-inflammatory factors TNF-α, IL-1β and IL-6 in hippocampus were increased in PS, DMSO+ PS and Nec-1+ PS groups ( P<0.05). Compared with PS group and DMSO+ PS group, the escape latency was significantly shortened, the time of staying at the original platform quadrant was prolonged, and the number of crossing the original platform was increased, the activation rate of microglia cells in the hippocampal dentate gyrus and necrosis rate of hippocampal neurons were decreased, the expression of RIPK1 and p-MLKL was down-regulated, and the contents of pro-inflammatory factors TNF-α, IL-1β and IL-6 in hippocampus were decreased in Nec-1+ PS group ( P<0.05). Conclusions:Necrostatin-1 pre-treatment can improve postoperative cognitive function in aged rats with chronic pain due to knee arthritis, and the mechanism may be related to inhibition of necrosis in hippocampal neurons and reduction of neuroinflammation.

Objective:To evaluate the efficacy of daily use of a test emollient combined with topical glucocorticoids applied at the weekend for delaying the recurrence of atopic dermatitis (AD) in children during the maintenance period.Methods:A randomized, blank-controlled, multicenter clinical study was conducted in children with moderate AD from Beijing Children′s Hospital, Capital Medical University, Children′s Hospital of Chongqing Medical University and Shenzhen Children′s Hospital from March 2021 to February 2022. A total of 127 children aged 0 - 12 years with moderate AD were treated with topical glucocorticoids combined with emollients during the run-in period, 112 out of them achieved the investigator′s global assessment (IGA) score ≤ 1 point, and then the 112 patients were randomly divided into a test group (56 cases) and a control group (56 cases) at a ratio of 1∶1. Patients in the test group received treatment with a test emollient twice a day in combination with topical glucocorticoids applied at the weekend, and those in the control group were only treated with topical glucocorticoids at the weekend. Patients in the two groups were followed up at baseline, week 2 (± 3 d), week 4 (± 5 d), and week 12 (±7 d), as well as at the time of AD relapse, and the effect of the test emollient on the remission rate of AD in children during the maintenance period was evaluated, so were its effects on the dosage of topical glucocorticoids, pruritus, sleep, and skin pH. The occurrence of treatment-related adverse events was evaluated and recorded at the same time. Study endpoints were defined as AD relapse during the maintenance period, end of 12-week follow-up, or occurrence of serious adverse events. Comparisons of efficacy indicators between groups were conducted by using chi-square test, Kaplan-Meier survival analysis, Satterthwaite t′ test and Mann-Whitney U test. Results:In the full-analysis set, 45 (80.36%) patients with AD maintained remission in the test group (56 cases) and 30 (53.57%) in the control group (56 cases), and the remission rate difference between the two groups was 26.79% (95% confidence interval [ CI]: 10.09%, 43.49%; χ2 = 9.11, P = 0.003) ; the 12-week follow-up during the maintenance period showed that the time to first relapse was 75.05 ± 25.07 days in the test group, which was significantly longer than that in the control group (49.55 ± 33.92 days, t′ = 4.52, P < 0.001). At the study endpoint, the test group showed significantly decreased AD disease severity score (eczema area and severity index [EASI] score: 0.00 [0.00, 1.20] points vs. 0.60 [0.00, 4.00] points), pruritus visual analog scale (VAS) score (0.00 [0.00, 2.00] points vs. 2. 00 [0.00, 10.00] points), and sleep VAS score (0.00 [0.00, 0.00] points vs. 1.00 [0.00, 4.00] points) compared with the control group ( Z = -2.77, 2.43, 3.48, P = 0.006, = 0.015, < 0.001, respectively), while there was no significant difference in the pH value at the lesional sites between the test group and control group ( t = 0.97, P = 0.335). For the group aged 0 - 2 years, the average daily glucocorticoid dosage at the weekend in AD children during the maintenance period was significantly lower in the test group than in the control group ( Z = -1.97, P = 0.049) ; for the group aged >2 - 12 years, there was no significant difference in the average daily glucocorticoid dosage at the weekend between the two groups ( Z = -0.25, P = 0.802). During the study period, no significant difference was observed in the incidence of treatment-related adverse events between the test group (2/56, 3.57%) and control group (3/56, 5.36%; P = 1.000), and no serious adverse events occurred. Conclusion:Compared with the weekend treatment with topical glucocorticoids alone, the daily use of the test emollient combined with topical glucocorticoids at the weekend could markedly improve the remission rate of AD, prolong the time to relapse, and reduce the disease severity at relapse in children with AD during the maintenance period, which provides a new option for maintenance treatment of children with AD.

Objective:To evaluate the role of brain-derived neurotrophic factor (BDNF)/tropomyosin-related kinase B (TrkB) signaling pathway in pre-injection of young rat plasma-induced reduction of sevoflurane-caused cognitive dysfunction in aged rats.Methods:Eighty SPF healthy male Sprague-Dawley rats, aged 18 months, weighing 550-650 g, were divided into 4 groups ( n=20 each) using a random number table method: control group (group C), sevoflurane anesthesia group (group S), young rat plasma group (group Y) and BDNF/TrkB signaling pathway inhibitor K252a group (group K). The plasma 100 μl obtained from 3-month-old young rats was injected via the tail vein in group Y and group K, while the equal volume of normal saline was given via the tail vein in group C and group S, twice a week, for 4 weeks.In S, Y and K groups, 3% sevoflurane was inhaled for 3 h starting from the end of treatment, and BDNF/TrkB signaling pathway inhibitor K252a was injected via the tail vein before anesthesia in group K. The open field test and Morris water maze test were performed at 3 days after anesthesia to assess the spontaneous motor ability and cognitive function.Then the rats were sacrificed, and the hippocampal tissues were isolated for determination of the expression of BDNF, phosphorylated TrkB (p-TrkB), postsynaptic dense protein-95 (PSD-95) and synaptic vesicle protein (SYN) (by Western blot), dendritic length and dendritic ridge density of neurons in hippocampal CA1 area (by Golgi staining), and the number of synapses and length of synaptic active area (with a transmission electron microscope). Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the expression of p-TrkB, BDNF, PSD-95 and SYN was down-regulated, and the dendritic length, dendritic ridge density, the number of synapses and length of synaptic active area were decreased in group S ( P<0.05). Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, the expression of p-TrkB, BDNF, PSD-95 and SYN was up-regulated, and the dendritic length, dendritic ridge density, the number of synapses and length of synaptic active area were increased in group Y ( P<0.05). Compared with group Y, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the expression of p-TrkB, BDNF, PSD-95 and SYN was down-regulated, and the dendritic length, dendritic ridge density, the number of synapses and length of synaptic active area were decreased in group K ( P<0.05). Conclusions:The mechanism by which pre-injection of young rat plasma reduces sevoflurane-induced cognitive dysfunction is related to activation of BDNF/TrkB signaling pathway and improvement in synaptic plasticity in the hippocampus of aged rats.

Objective:To investigate the whole genome of SARS-CoV-2 causing COVID-19 in Rongcheng city of Shandong Province in May 2022 and to further analyze the nucleotide and amino acid variations for source tracing.Methods:High-throughput sequencing was used to sequence the SARS-CoV-2 genome in 15 nasopharyngeal swab samples from COVID-19 cluster infections and three environmental samples related to an aquatic product import company. Whole-genome sequence splicing, variant site analysis and sequence typing were performed on the raw sequencing data using virus sequence and variant analysis software. A phylogenetic tree was constructed by evolutionary analysis software. Epidemiological investigation was used to trace the potential source of infection.Results:Thirteen whole genome sequences of SARS-CoV-2 with the length ranging from 29 653 bp to 29 780 bp were successfully obtained from the nasopharyngeal swab samples. The average sequencing depth was 1 756-6 565 X and the genome coverage was 99.20%-99.63%. The results of Pangolin typing showed that the 13 genomes belonged to the VOC/Gamma (P.1.15) evolutionary branch. Compared with the Wuhan reference strain (NC_045512.2), the 13 genome sequences had 40-41 nucleotide mutation sites. There were 23-24 amino acid variation sites in seven protein domains (ORF1a, ORF1b, S, ORF3a, ORF8, ORF9b and N proteins). Evolutionary analysis showed that the viral sequence was grouped to the same subclade as the reference strain from Argentina (EPI_ISL_4082233).Conclusions:In this study, the whole genome sequences of 13 Gamma variant strains were obtained from COVID-19 cluster infections associated with imported cold-chain aquatic products in Rongcheng city, and the imported seafood from South America in 2021 was found to be the source of the virus in a timely manner. This study provided reference for the SARS-CoV-2 variant analysis and case tracing and also suggested that the survival and transmission ability of SARS-CoV-2 on the surface of cold-chain products should not be underestimated and needed further investigation.

Objective:To evaluate the role of N-methyl-D-aspartate receptors (NMDA receptors) in sevoflurane anesthesia-caused necroptosis in hippocampal neurons of aged mice.Methods:Ninety clean-grade healthy male C57BL/6 mice, aged 18 months, weighing 27-30 g, were divided into 3 groups ( n=30 each) using a random number table method: control group (group C), sevoflurane anesthesia group (group S) and sevoflurane anesthesia plus NMDA receptor antagonist memantine hydrochloride group (group S+ M). Mice inhaled 3% sevoflurane for 2 h for 3 consecutive days in S group and S+ M group, and memantine hydrochloride 20 mg/kg was intraperitoneally injected at 1 h before each inhalation of sevoflurane in S+ M group.Mice only inhaled pure oxygen for 2 h in group C. Ten mice of each group were selected on 1 day before anesthesia and 3 and 7 days after anesthesia to perform Morris water maze test.The mice were sacrificed immediately after Morris water maze test, and hippocampus was removed for microscopic examination of pathological changes (with a light microscope) and for determination of the necroptosis rate of neurons and cytoplasmic free calcium concentration([Ca 2+ ] i)(by flow cytometry), and expression of NMDA receptor subtypes GluN2A, GluN2B and receptor-interacting protein kinase 1 (RIP1) (by Western blot). Results:Compared with group C, the escape latency was significantly prolonged, and the frequency of crossing the original platform was decreased, and the [Ca 2+ ] i and neuronal necroptosis rate in the hippocampus were increased at each time point after anesthesia, and the expression of GluN2A, GluN2B and RIP1 was up-regulated( P<0.05), and the pathologic changes were accentuated in S group and S+ M group.Compared with group S, the escape latency was significantly shortened, and the frequency of crossing the original platform was increased, and the [Ca 2+ ] i and neuronal necroptosis rate in the hippocampus were decreased at each time point after anesthesia, and the expression of GluN2A, GluN2B and RIP1 was down-regulated ( P<0.05), and the pathologic changes were attenuated in group S+ M. Conclusions:NMDA receptors are involved in the process of cognitive dysfunction induced by sevoflurane anesthesia in aged mice, and the mechanism may be related to the promotion of necrptosis in hippocampal neurons.

Objective:To evaluate the role of RhoA/ROCK2 signaling pathway in multiple exposures to sevoflurane-induced long-term cognitive impairment in neonatal rats.Methods:Sixty SPF healthy neonatal Sprague-Dawley rats of either sex, aged 6 days, weighing 12-20 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), multiple exposures to sevoflurane group (group S) and RhoA/ROCK2 signaling pathway inhibitor Y-27632 group (group Y). Group S and group Y inhaled 3% sevoflurane for 2 h at days 6, 7 and 8 after birth.In group Y, Y-27632 5 mg/kg was intraperitoneally injected before sevoflurane anesthesia.The spontaneous activity was evaluated by open field test on day 35 after birth.The cognitive function was detected by Morris water maze test at day 36 after birth.The rats were sacrificed after Morris water maze test, and the hippocampal tissues were isolated for determination of the apoptosis rate of hippocampal neurons and cytoplasmic calcium concentration ([Ca 2+ ] i) (by flow cytometry) and expression of phosphorylated RhoA (p-RhoA), ROCK2 and cleaved-caspase-3 (by Western blot) and for microscopic examination of the ultrastructure of hippocampal neurons (with a transmission electron microscope). Results:There was no significant difference in movement speed, distance and time of stay in the open field center in the open field test among the three groups ( P>0.05). Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the apoptosis rate of hippocampal neurons and [Ca 2+ ] i were increased, the expression of p-RhoA, ROCK2 and cleaved-caspase-3 was up-regulated ( P<0.05), and the pathological injury to hippocampal neurons was found in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, the apoptosis rate of hippocampal neurons and [Ca 2+ ] i were decreased, the expression of p-RhoA, ROCK2 and cleaved-caspase-3 was down-regulated ( P<0.05), and the pathological injury to hippocampal neurons was attenuated in group Y. Conclusions:The mechanism by which multiple exposures to sevoflurane induces long-term cognitive impairment is related to activation of RhoA/Rock2 signaling pathway and induction of apoptosis rate of hippocampal neurons in neonatal rats.

OBJECTIVE To study the improvement effects and mechanism of gentian iridoids （GI）on lipid accumulation and inflammation in non -alcoholic fatty liver disease （NAFLD）model. METHODS Human hepatoma HepG 2 cells induced by free fatty acids（0.5 mmol/L）were selected as NAFLD cell models . The effects of GI at different concentrations （0.125，0.25，0.5，1，2，4 mg/mL）on HepG 2 cell viability were investigated . HepG2 cells were divided into control group ，model group and low ，medium and high concentration groups of GI （0.25，0.5 and 1 mg/mL）. Except for control group ，0.5 mmol/L free fatty acid was added in other groups for modeling .After 24 h of culture ，the formation of lipid droplets was observed in HepG 2 cells；the content of triglyceride（TG）was detected . The levels of aspartate aminotransferase （AST），alanine aminotransferase （ALT）and inflammation indexes [interleukin-1 β（IL-1 β），IL-6，tumor necrosis factor α（TNF-α）] in cell supernatant were determined . The mRNA expressions of sterol -regulatory element binding protein （SREBP-1c）and fatty acid synthase （FAS）as well as the expression of nuclear factor -κB（NF-κB）related proteins [NF-κB inhibitor protein α（IκBα），NF-κB，phosphorylated NF -κB（p-NF-κB）] were also detected . RESULTS After treated with different concentrations of GI ，the cell viability of HepG 2 cells was improved significantly（P＜0.01）. Compared with model group ，the number of red lipid droplets in the cells of each concentration group of GI was significantly reduced ，the nuclear atrophy was not obvious ，and the size was normal ；the content of TG （except for low concentration group ），and the levels of ALT ，AST，IL-1β，IL-6（except for low concentration group ）and TNF - α in cell supernatant were all decreased significantly ；mRNA expression of SREBP -1c and FAS and the phosphorylation level of NF -κB were also decreased significantly （P＜0.05 or P＜0.01）；the protein expression of I κBα was increased significantly （P＜0.01）. CONCLUSIONS GI can reduce the lipid accumulation and inflammatory response of NAFLD cell model induced by free fatty acids，and its mechanism may be related to the inhibition of SREBP-1c/FAS and NF -κB signaling pathway .