Objective To investigate optimal method of establishing hypoxia/reoxygenation(H/R) model of H9c2 cell by using hypoxia/anoxic workstation under different conditions in hypoxia.Methods H9c2 cell was placed into hypoxia/anoxic workstation and simultaneously cultured with complete medium, glucose-free DMEM and acidic hypoxic solution for 1,2,4,6 and 8 h respectively, and then reoxygenated with complete medium for 1 h in normoxic incubator.The level of ROS was measured by flow cytometry.The cell viability was detected by MTT assay.The cellular morphology was observed by inverted microscope.Results With the extension of cell hypoxia time, there were no significant differences in the ROS level and cell viability in complete medium-and glucose-free DMEM-treated H/R groups compared with control group(P<0.05).There was no obvious morphologic change observed with inverted microscope, either.Nevertheless, when H9c2 cells were treated with acidic hypoxic solution in hypoxia, the ROS level continuously increased and the cell viability decreased with the extension of cell hypoxia time ( P<0.01 ).Since H:1 h/R:1 h, some of the cells shrunk and a few necrotic cells floated in the media under the inverted microscope , and the damage was aggravated with the extension of hypoxia time.After the cells were exposed in hypoxia for 8 h, they wrinkled to be round and a large number of floating necrotic cells were observed.When the cells were reoxygenated for 1 h, the cytomembrane was not smooth and there were still a few necrotic cells floating in culture dish .Conclusion The H9c2 cell H/R model with good repeatability can be established successfully by using hypoxia/anoxic workstation combining with acidic hypoxic solution.

A robust,selective and highly sensitive chemiluminescent (CL) platform for protein assay was presented in this paper.This novel CL approach utilized rolling circle amplification (RCA) as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier.Typically,the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence.This aptamer-primer sequence was then employed as the primer of RCA.Based on this design,a number of the biotinylated probes and streptavidin-horseradish peroxidase (SA-HRP) were captured on the plate,and the CL signal was amplified.In summary,our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized,and devoid of light source.Therefore,this new technique will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases,by taking advantages of high sensitivity and selectivity.

Objective Cancer-related fatigue (CRF) is a key in the management of cancer patients' clinic syptoms.This article investigated the status quo of nurses' knowledge and attitude towards CFR.Methods THe method of cross-sectional survey and questionnaire was used to investigate the knowledge and attitude towards CRF among nurses from related departments of three Grade III hospitals in Nanjing.Results 142 nurses answered the questionnaire.The average correct rate was 76.25%, among which nurses from the oncology department had better congition rate than nurses from other medical and surgical departments (84.3%, 75.98%, 79.57%) , representing significant difference (P<0.05).64.79% of the nurses found the relatives of cancer patients and nurses often fail to understand cancer patients;complaint of fatigue, 76.76% of nurses assumed there is lack of communication in fatigue between patients and medical staff.94.36% of nurses agreed medical institutions should strengthen the management of CRF.Conclusion At present, the clinical nurses have inadequate knowledge about CRF, which should be enhanced in future work.

A robust, selective and highly sensitive chemiluminescent （CL） platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification （RCA） as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase （SA-HRP） were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique .will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.

Objective To study the influence of residual methylene blue after plasma viral inactivation on the human immune cell function by using the peripheral blood mononuclear cell(PBMC) .Methods PBMC were isolated by adopting the Ficoll‐Hypaque density gradient centrifugation method and co‐cultured for 72 h in presence of specific T cell stimulating factors(Anti‐CD3/28 and Anti‐CD28) ,with or without different concentration of methylene blue .The culture supernatant was collected and detected the cyto‐kines secretion situation by ELISA .After 66 h culture ,CCK‐8 dye was added and continueously cultured for 4-6 h ,the prolifera‐tion was determined at A450 .Results The high‐concentration doses of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) had signifi‐cantly inhibiting effect on the proliferation of PBMC stimulated by Anti‐CD3/28(P< 0 .01) ,its OD value was decreased from 0 .897 ± 0 .385 to 0 .632 ± 0 .334 ,0 .524 ± 0 .254 and 0 .445 ± 0 .287 respectively ,showing certain dose dependent effect .The high concentrations of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) could down‐regulate interleukin(IL)‐17a ,IL‐10 and interferon (IFN)‐γ secreted by anti‐CD28 induced PBMC ,moreover showing a dose dependent effect .1 .25 ,2 .5 ,5 μmol/L methylene blue af‐fected the IL‐17a level secreted by PBMC from (406 ± 57)pg/mL descending to (276 ± 38) ,(192 ± 31) ,(134 ± 24)pg/mL respec‐tively ;affected PBMC to secrete IL‐10 ,its level was reduced from (184 ± 15) pg/mL to (132 ± 13) ,(110 ± 12) ,(42 ± 8)pg/mL ;af‐fected PBMC to secrete IFN‐γ,its level was deduced from (4 512 ± 187)pg/mL to (2 876 ± 143) ,(2 234 ± 153) ,(1 988 ± 112)pg/mL respectively .Conclusion High concentrations of methylene blue (≥1 .25 μmol/L ) has the significant inhibiting effect on the proliferation and cytokine secretion functions of PBMC .In other words ,the residual methylene blue concentration in viral inactiva‐tion plasma (≤0 .33 μmol/L) has no obvious effect on the immune function of PBMC ,but whether this concentration of methylene blue having the effect on human pure T cell immune function needs to be further evaluated and studied .

Lysine methyltransferase 2A( KMT2A) rearrangement (KMT2A-r) is a high-risk gene subtype of acute lymphoblastic leukemia (ALL) in children with a high relapse rate, poor prognosis and suboptimal response to conventional chemotherapy.Improving the treatment and prognosis of childhood KMT2A-r ALL is an urgent problem.In recent years, with an in-depth understanding of the mechanism of KMT2A-r, more accurate risk stratification of KMT2A-r ALL has been carried out, and great progress has also been made in its immune and targeted therapy.In this article, the genetic and biological characteristics, risk stratification, treatment strategies and prognosis of children KMT2A-r ALL were reviewed in order to provide theoretical support for clinical work and future research.

Objective:To analyze the research hotspots and trends of the application of Kirkpatrick model in nursing education through bibliometrics analysis, so as to provides ideas for follow-up researchers to carry out research in related fields.Methods:Relevant literatures in CNKI, Wanfang, VIP and PubMed were retrieved from the establishment of databases until August 27, 2022. CiteSpace software was used to draw the literature topic clustering map, BICOMB was used to generate the co-occurrence matrix of bibliographic data, and gCLUTO software was used to make the visual hill map and analyze the literature hotspots.Results:A total of 230 literatures were included, including 145 in Chinese and 85 in English. Since 2016, the number of published papers had been on the rise. The papers were published in 134 journals at home and abroad, and the coauthor rate was 95.65% (220/230), and the funded papers accounted for 25.65% (59/230). The hot topic of Chinese literature research was the construction of training evaluation index system and training effectiveness evaluation. The research objects mainly included specialized nurses and in-service nurses.The research hotspot of English literature was nursing education and training. The research objects were specialized nurses and undergraduate students.Conclusions:The application of the Kirkpatrick model in the field of specialized nurse training is still in its infancy, and it needs to be extended to more specialist fields in the future. In addition, researchers should further explore the construction and application of a training evaluation index system based on the Kirkpatrick model in the future, and promote the horizontal development of training

Objective:To explore the rehabilitation effect of multi-factor intervention based on the Finnish model of prevention of cognitive impairment in the elderly on patients with cognitive impairment after first-episode stroke, and to provide reference for rehabilitation nursing of cognitive impairment after stroke.Methods:The quasi-experiment research scheme was adopted and convenience sampling method was used to select participants with first-episode stroke cognitive impairment hospitalized in the General Hospital of Tianjin Medical University Airport Site. The 50 patients admitted from January to June 2022 were selected as the control group, and 50 patients admitted from July to December 2022 were selected as the intervention group. The control group received routine rehabilitation nursing and health education, and the intervention group received the Finnish model of prevention of cognitive impairment in the elderly on patients before discharge on the basis of the control group. The Mini-Mental State Examination (MMSE) and Health Education Compliance Assessment Scale for Stroke Patients were used to evaluate the changes of overall cognitive function and rehabilitation compliance before intervention, 3 and 6 months after intervention.Results:The final control group included 49 cases, including 35 males and 14 females, aged (64.67 ± 7.47) years old; the intervention group included 50 cases, 32 males and 18 females, aged (66.68 ± 8.75) years old. Before intervention, there were no significant differences in overall cognitive function and compliance of rehabilitation score ( P>0.05). At 3 and 6 months after intervention, the overall cognitive function score, the total score on compliance of rehabilitation, dimension scores of diet compliance, exercise rehabilitation compliance and health behavior compliance of the intervention group were (26.36±2.36) , (125.96 ± 13.80) , (23.30 ± 5.26) , (27.72 ± 4.46) , (43.66 ± 6.80) and (27.26 ± 3.71) , (152.44 ± 9.06) , (30.12 ± 6.42) , (33.32 ± 3.02) , (52.36 ± 4.70) , respectively. They were higher than the control group (24.04 ± 4.50) , (116.67 ± 10.26) , (19.31 ± 3.95) , (25.29 ± 3.45) , (40.59 ± 4.33) and (24.27 ± 4.33) , (138.92 ± 16.71) , (24.20 ± 4.48) , (30.00 ± 5.53) , (47.65 ± 8.03) , and the differences had statistical significance ( t values were -5.31- -2.67, all P<0.05). According to the variance analysis of repeated measurement, intergroup and time factor, the interaction between groups and time had significant impact on general cognitive function score, the total score of rehabilitation compliance, the dimension scores of diet, exercise rehabilitation and health behavior compliance ( Fgroup values were 8.33-18.08, Ftime values were 135.71-944.69, Finteraction values were 5.46-27.30, all P<0.05) . Time factor had significant impact on patient medication adherence score ( Ftime=206.23, P<0.05) . Conclusions:Multi-factor intervention based on the Finnish model of prevention of cognitive impairment in the elderly can improve the overall cognitive function and rehabilitation compliance of patients with cognitive impairment after first-episode stroke.

Objective:To evaluate the efficacy of esketamine-based anesthesia in lumbar spine surgery.Methods:Ninety-four patients of both sexes, aged 18-64 yr, with body mass index of 18.5-29.9 kg/m 2, of American Society of Anesthesiologists Physical Status classification ⅠorⅡ, scheduled for elective lumbar posterior decompression bone grafting fusion internal fixation under general anesthesia from June 2022 to December 2022, were divided into control group(group C) and esketamine group(group K) using a random number table method, with 47 cases in each group. Midazolamm, sufentanil, etomidate and cisatracurium were intravenously injected for anesthesia induction in both groups, and esketamine 0.5 mg/kg was intravenously injected on this basis in group K. Propofol and remifentanil were intravenously infused to maintain anesthesia, and cisatracurium besylate was intermittently injected to maintain muscle relaxation in both groups, and esketamine 0.25 mg·kg -1·h -1 was intravenously infused on this basis in group K. The patients were connected to an analgesic pump for patient-controlled intravenous analgesia at 10 min before the end of surgery, and flurbiprofen axetil 50 mg was intravenously injected for rescue analgesia when the numeric rating scale score >4. The time of first pressing the analgesia pump, effective pressing times of the analgesia pump within 48 h after operation and requirement for rescue analgesia were recorded. The initial dose of remifentanil, cumulative amount of remifentanil used during operation, time of tracheal extubation, and adverse reactions within 48 h after surgery were recorded. Results:Compared with group C, the cumulative use of remifentanil during operation was significantly reduced, the time of first pressing the self-control button of the analgesia pump after surgery was prolonged, the pressing times of the analgesia pumps were decreased( P<0.05), and no significant change was found in terms of the initial dose of intraoperative remifentanil, rate of postoperative rescue analgesia, time of extubation, and incidence of adverse reactions after surgery in group K( P>0.05). Conclusions:Esketamine-based anesthesia can reduce the amount of intraoperative opioids, delay the time of postoperative pain and reduce the early postoperative pain when used for lumbar spine surgery.

Objective:To retrieve the relevant evidence of the application of virtual reality technology in the cognitive rehabilitation of stroke patients and conduct evaluation and summary of the evidence to provide a reference for formulating norms of cognitive rehabilitation of stroke based on virtual reality technology.Methods:Using evidence-based nursing methods, according to the "6S" model system to search guidelines, expert consensus, random controlled trail (RCT) , systematic reviews and other evidence of the use of virtual reality technology in the cognitive rehabilitation training of stroke patients on United States National Guidelines Network, National Institute for Health and Care Excellence, Registered Nurses' Association of Ontario, Scottish Intercollegiate Guidelines Network, Cochrane Library, Joanna Briggs Institute (JBI) Evidence-Based Health Care Database, PubMed, Web of Science, Springerlink, Science Direct, National Stroke Foundation, China Guidance Network, Medlive, CNKI, Wanfang Database and VIP Database. The retrieval period was from January 2015 to December 2020. Using Appraisal of Guidelines for Research and Evaluation Ⅱ (AGREE Ⅱ) , Assessment of Multiple Systematic Reviews 2 (AMSTAR 2) and JBI literature quality evaluation tools, two researchers independently evaluated the quality of the literature and the level of evidence.Results:Finally, 17 articles were included, including 6 guidelines, 4 systematic reviews and 7 RCT studies, summarizing 20 evidences of the application of virtual reality technology in cognitive rehabilitation of stroke patients.Conclusions:This research has initially formed a summary and evaluation of the evidence for the application of virtual reality technology in cognitive rehabilitation of stroke patients, in order to provide a reference for the formulation of practical norms of cognitive rehabilitation technology based on virtual reality and improve the effect of cognitive rehabilitation for stroke patients.