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.

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.

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 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 investigate the association between alcohol drinking and all-cause death in patients with ischemic stroke. METHODS: Between January, 2010 and July, 2018, consecutive patients with first-episode ischemic stroke admitted in the West China Hospital, Sichuan University were enrolled, and all the patients were followed up every 3 months.Chi-square test was used to compare the differences in the baseline characteristics between the pre-stroke drinkers and the nondrinkers.The Cox regression model was used to analyze the effects of drinking status, drinking years, drinking frequency, average single alcohol intake before stroke and drinking status during the follow-up period on the mortality of patients after discharge. RESULTS: A total of 855 patients with ischemic stroke were enrolled, and deaths occurred in 140 of these patients.Chi-square test showed significant differences in gender ( < 0.001), weekly physical exercise time (=0.035), smoking ( < 0.001), and heart disease ( < 0.001) between the pre-stroke drinkers and nondrinkers.Multivariate Cox regression analysis showed that drinking during the follow-up period (=0.001), drinking for less than 28 years before stroke (=0.035) and a moderate drinking frequency (5 to 20 times per month for males and 4 to 9 times per month for females; =0.030) were associated with a lowered risk of death after discharge.No significant effects of pre-stroke drinking status or average single alcohol intake were found on death after ischemic stroke. CONCLUSIONS The drinking years and drinking frequency before stroke and drinking status during the follow-up period are related to the all-cause mortality in patients with ischemic stroke.Investigations of the more specific variables of drinking behaviors during the follow-up period are needed to further clarify the association between drinking and death after ischemic stroke.
Alcohol Drinking ; adverse effects ; Brain Ischemia ; etiology ; China ; Female ; Follow-Up Studies ; Humans ; Male ; Risk Factors ; Stroke ; etiology