Objective To establish models for real-time dynamic monitoring of intracellular cytoplasmic adenosine triphosphate(ATP)and mitochondrial ATP levels in cells in order to study the changes in metabolic processes.Methods The lentiviral plasmids of the cytoplasmic chemogenetic green fluorescent protein(GFP)ATP probe(Chemo-G)and those of the mitochondrial-localized chemogenetic GFP ATP probe(mito-Chemo-G)were constructed before being transfected into HEK293T together with the helper plasmids,respectively.Chemo-G and mito-Chemo-G lentiviruses were obtained.HeLa cells were infected with the lentivirus.Puromycin resistance selection and flow cytometry cell sorting were employed to identify and isolate the infected HeLa cells.The growth and GFP expressions of HeLa cells were observed.A live-cell imaging system was used for continuous imaging of the cells,with stimuli added at specific time points to alter intracellular ATP levels to observe changes in the fluorescence intensity of the ATP probe.Results Lentiviral plasmids containing Chemo-G and mito-Chemo-G sequences were constructed.Two cell lines which could stably express Chemo-G and mito-Chemo-G were established that exhibited strong growth and accurate intracellular fluorescence localization.Live-cell imaging revealed that after the addition of 2-deoxy-D-glucose(2-DG)into HeLa-Chemo-G,the fluorescence resonance energy transfer(FRET)/GFP ratio showed a decrease that was partially reversed by the addition of glucose.The FRET/GFP ratio increased after histamine stimulation,but rapidly decreased after the addition of oligomycin.Conclusion The Chemo-G and mito-Chemo-G lentiviral vectors and stably transfected cell lines HeLa-Chemo-G and HeLa-mito-Chemo-G are constructed,which provides reliable experimental models for studying cellular metabolism and changes in intracellular ATP levels.

OBJECTIVES: To report the development, validation, and findings of the Multi-dimensional Attention Rating Scale (MARS), a self-report tool crafted to evaluate six-dimension attention levels. METHODS: The MARS was developed based on Classical Test Theory (CTT). Totally 202 highly educated healthy adult participants were recruited for reliability and validity tests. Reliability was measured using Cronbach's alpha and test-retest reliability. Structural validity was explored using principal component analysis. Criterion validity was analyzed by correlating MARS scores with the Toronto Hospital Alertness Test (THAT), the Attentional Control Scale (ACS), and the Attention Network Test (ANT). RESULTS: The MARS comprises 12 items spanning six distinct dimensions of attention: focused attention, sustained attention, shifting attention, selective attention, divided attention, and response inhibition.As assessed by six experts, the content validation index (CVI) was 0.95, the Cronbach's alpha for the MARS was 0.78, and the test-retest reliability was 0.81. Four factors were identified (cumulative variance contribution rate 68.79%). The total score of MARS was correlated positively with THAT (r = 0.60, P < 0.01) and ACS (r = 0.78, P < 0.01) and negatively with ANT's reaction time for alerting (r = -0.31, P = 0.049). CONCLUSIONS The MARS can reliably and validly assess six-dimension attention levels in real-world settings and is expected to be a new tool for assessing multi-dimensional attention impairments in different mental disorders.
Humans ; Adult ; Male ; Attention/physiology* ; Female ; Middle Aged ; Reproducibility of Results ; Young Adult ; Psychometrics

ObjectiveTo understand the whole genome characteristics and the information for genetic evolution in the two coxsackievirus A2 (CVA2) strains isolated from patients with herpangina in Shanghai, and to provide a scientific basis for the prevention and treatment of herpetic angina. MethodsTwo CAV2 strains isolated from patients with herpetic angina in Shanghai were performed whole genome sequencing and analysis for phylogenetics, nucleotide homology, and evolution. ResultsA phylogenetic analysis of the VP1 region revealed that the two Shanghai strains both belonged to CVA2 genotype D, with the highest homology to OL357660, a strain from Yunnan. The average nucleotide identity (ANI) of the whole genome between the two Shanghai strains was 98.88%, and the ANI of the whole genome comparisons to other CVA2 genotype D strains and CVA2 genotypes A-C strains ranged from 84.64% to 97.42% and from 79.21% to 84.20%, respectively. The two Shanghai strains had low homology in the 3D region compared to the existing CVA2 strains. The phylogenetic analysis and sliding window nucleotide similarity analysis indicated that the two Shanghai strains and the Yunnan OL357660 strain might constitute a new genetic lineage. ConclusionThe two CVA2 strains isolated for the first time in Shanghai are assigned to genotype D (GenBank: PQ130039 and PQ130040), which is identical to the existing subtype prevalent in China. As represented by the Shanghai strains, a new CVA2 genetic lineage is been identified. This study has enriched the data on genetic evolution and genetic variation of CVA2 in Shanghai, indicating the requirement to strengthen surveillance for the epidemiological pattern of CVA2.

Background and Objectives: Long-term pathological myocardial hypertrophy (MH) seriously affects the normal function of the heart. Dronedarone was reported to attenuate left ventricular hypertrophy of mice. However, the molecular regulatory mechanism of dronedarone in MH is unclear. Methods: Angiotensin II (Ang II) was used to induce cell hypertrophy of H9C2 cells.Transverse aortic constriction (TAC) surgery was performed to establish a rat model of MH.Cell size was evaluated using crystal violet staining and rhodamine phalloidin staining.Reverse transcription quantitative polymerase chain reaction and western blot were performed to detect the mRNA and protein expressions of genes. JASPAR and luciferase activity were conducted to predict and validate interaction between forkhead box O3 (FOXO3) and protein kinase inhibitor alpha (PKIA) promoter. Results: Ang II treatment induced cell hypertrophy and inhibited sirtuin 1 (SIRT1) expression, which were reversed by dronedarone. SIRT1 overexpression or PKIA overexpression enhanced dronedarone-mediated suppression of cell hypertrophy in Ang II-induced H9C2 cells. Mechanistically, SIRT1 elevated FOXO3 expression through SIRT1-mediated deacetylation of FOXO3 and FOXO3 upregulated PKIA expression through interacting with PKIA promoter. Moreover, SIRT1 silencing compromised dronedaronemediated suppression of cell hypertrophy, while PKIA upregulation abolished the influences of SIRT1 silencing. More importantly, dronedarone improved TAC surgery-induced MH and impairment of cardiac function of rats via affecting SIRT1/FOXO3/PKIA axis. Conclusions Dronedarone alleviated MH through mediating SIRT1/FOXO3/PKIA axis, which provide more evidences for dronedarone against MH.

INTRODUCTION: Emergency laparotomy (EL) is associated with high morbidity and mortality, often exceeding 10%. This study evaluated the impact of the EMergency Laparotomy Audit (EMLA) interdisciplinary perioperative pathway on patient outcomes, hospital costs and length of stay (LOS) within a single centre. METHOD: A prospective cohort study was conducted from August 2020 to July 2023. The intervention team included specialist clinicians, hospital administrators and an in-hospital quality improvement team. Patients who underwent EL were divided into a pre-intervention control group (n=136) and a post-intervention group (n=293), and an 8-item bundle was implemented. Propensity scoring with a 1:1 matching method was utilised to reduce confounding and selection bias. The primary outcomes examined were LOS, hospitalis-ation costs and surgical morbidity, while secondary outcomes included 30-day mortality and adherence to the intervention protocol. RESULTS: The utilisation of the EMLA perioperative care bundle led to a significant reduction in surgical complications (34.8% to 20.6%, P<0.01), a decrease in LOS by 3.3 days (15.4 to 12.1 days, P=0.03) and lower hospitalisation costs (SGD 40,160 to 30,948, P=0.04). Compliance with key interventions also showed improvement. However, there was no difference in 30-day mortality. CONCLUSION This study offers insights on how surgical units can implement systemic perioperative changes to improve outcomes for patients undergoing emergency laparotomy.
Humans ; Laparotomy/methods* ; Propensity Score ; Female ; Male ; Prospective Studies ; Length of Stay/statistics & numerical data* ; Middle Aged ; Hospital Costs/statistics & numerical data* ; Postoperative Complications/epidemiology* ; Aged ; Emergencies ; Perioperative Care/methods* ; Critical Pathways ; Singapore ; Adult

Objective:To analyze and evaluate the impact of balance training on patients with chronic obstructive pulmonary disease (COPD).Methods:The randomized controlled trials of balance training in COPD patients in PubMed, Embase, Web of science, the Cochrane Library, CNKI, Wanfang, VIP, and Chinese Biomedical Literature Database were searched. The retrieval time was from the establishment of the database to November 20, 2022. A Meta-analysis was conducted using RevMan5.3 on balance ability, exercise ability, quality of life, and balance confidence in patients with COPD.Results:A total of 432 patients were included in 7 studies. The results of the Meta-analysis showed that compared with other training, balanced training could improve the balance ability of patients [Timed Up and Go Test: mean difference ( MD)=-2.19, 95% CI (-2.86, -1.52); P<0.001, Berg Balance Scale: MD=4.02, 95% CI (0.78, 7.27); P=0.020, the Balance Evaluation Systems Test: MD=7.83, 95% CI (0.13, 15.52); P=0.050], exercise ability [Six-Minute Walk Test: standard mean difference ( SMD)=0.21, 95% CI (0.01, 0.40); P=0.040] and quality of life [COPD Assessment Test and St. George′s Respiratory Questionnaire: SMD=-0.54, 95% CI (-1.08, -0.01); P=0.050], but balance training did not effectively enhance patients′ activity balance confidence [Activities-Specific Balance Confidence Scale: MD=9.34, 95% CI (-0.98, 19.66); P=0.080]. Conclusion:Balance training can improve the balance ability, exercise ability, and quality of life of patients with COPD, but the impact on patients′ activity balance confidence needs to be further verified.

Chronic psychological stress can promote vascular diseases, such as hypertension and atherosclerosis. This study aims to explore the effects and mechanism of chronic psychological stress on aortic medial calcification (AMC). Rat arterial calcification model was established by nicotine gavage in combination with vitamin D₃ (VitD₃) intramuscular injection, and rat model of chronic psychological stress was induced by humid environment. Aortic calcification in rats was evaluated by using Alizarin red staining, aortic calcium content detection, and alkaline phosphatase (ALP) activity assay. The expression levels of the related proteins, including vascular smooth muscle cells (VSMCs) contractile phenotype marker SM22α, osteoblast-like phenotype marker RUNX2, and endoplasmic reticulum stress (ERS) markers (GRP78 and CHOP), were determined by Western blot. The results showed that chronic psychological stress alone induced AMC in rats, further aggravated AMC induced by nicotine in combination with VitD₃, promoted the osteoblast-like phenotype transformation of VSMCs and aortic ERS activation, and significantly increased the plasma cortisol levels. The 11β-hydroxylase inhibitor metyrapone effectively reduced chronic psychological stress-induced plasma cortisol levels and ameliorated AMC and aortic ERS in chronic psychological stress model rats. Conversely, the glucocorticoid receptor agonist dexamethasone induced AMC, promoted AMC induced by nicotine combined with VitD₃, and further activated aortic ERS. The above effects of dexamethasone could be inhibited by ERS inhibitor 4-phenylbutyrate. These results suggest that chronic psychological stress can lead to the occurrence and development of AMC by promoting glucocorticoid synthesis, which may provide new strategies and targets for the prevention and control of AMC.
Rats ; Animals ; Glucocorticoids/metabolism* ; Rats, Sprague-Dawley ; Nicotine/metabolism* ; Hydrocortisone/metabolism* ; Muscle, Smooth, Vascular ; Dexamethasone/metabolism* ; Vascular Calcification/metabolism* ; Myocytes, Smooth Muscle/metabolism* ; Cells, Cultured

No consensus on standardized technique of enterostomy creation has been made meanwhile high heterogeneity of surgical procedure exists in 'stoma creation' chapters of textbooks or atlases of colorectal surgery. The present article reviews the anatomy of tendinous aponeurotic fibers which is crucial for abdominal wall tension and integrity. Through empirical practice we hypothesize a procedure of enterostomy creation basied on abdominal wall tension plus anchor suture for fascia fixation which could theoretically decrease short-term stoma complication rates and long-term parastomal hernia rates. Surgical techniques are as followed: (1) preoperative stoma site mark for de-functioning ileostomy should be positioned at the lateral border of rectus abdominis muscle (RAM) to decrease the difficulty of stoma reversal and for permanent colostomy should be placed overlying the RAM to promote adhesion; (2)Optimal circular removal or lineal opening of skin, and avoid dissection of subcutaneous tissue; (3) Lineal dissection of natural strong fascia (rectus sheath) at stoma site and blunt separation of muscular fibers. The tunnel of the fascia should be made with appropriate size without undue tension. To prevent the formation of dead space, additional suturing at fascia layer is unnecessary. (4) Anchor suture for fascia fixation at two ends of fascia opening could be considered to avoid delayed fascia disruption and parastomal hernia. (5) After pull-through of ileum or colon loop, 4-8 interrupted seromuscular sutures could be placed to attach loop to skin. For ileostomy, self-eversion of mucosa can be successful in vast majority of cases and a Brooke ileostomy is not necessary. The efficacy and safety of this procedure should be tested in future trials.
Humans ; Abdominal Wall/surgery* ; Surgical Stomas/adverse effects* ; Enterostomy ; Incisional Hernia ; Fascia

Objective:To systematically review the effects of health coaching on patients with chronic obstructive pulmonary disease (COPD) .Methods:CNKI, Wanfang Database, China Biology Medicine disc, VIP Database, PubMed, Embase, Cochrane Library, and Web of Science were searched for randomized controlled trials concerning the health coaching in COPD patients which were published up to January 10, 2022. Two researchers independently screened the retrieved literature, extracted data, and evaluated the quality of the included literature according to the Cochrane 5.1.0 handbook. Meta-analysis was performed with RevMan 5.3.Results:Finally, 10 studies including 1 797 COPD patients were included. Meta-analysis showed that compared with the routine care group, patients in the health coaching group experienced reduced readmission rates ( OR=0.42, 95% CI: 0.29-0.60, P<0.01), alleviated depression ( MD=-0.51, 95% CI: -0.98--0.04, P=0.03), improved disease awareness ( MD=3.83, 95% CI: 0.97-6.69, P=0.009), and enhanced self-efficacy ( SMD=0.32, 95% CI: 0.09-0.55, P=0.006). The quality of life was analyzed in subgroups according to different research tools, and the results revealed that there was a statistically significant difference in the combined effect size among the subgroups ( P<0.05) ; however, it showed no obvious advantage in improving exercise tolerance and reducing anxiety. Conclusions:Health coaching can effectively reduce the readmission rate, alleviate the depression, improve the disease awareness, and enhance the self-efficacy and quality of life among COPD patients. However, there is a lack of evidence for its effect on exercise endurance and anxiety, and in-depth research is needed to verify it.