BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.

Objective To explore the relationship between education level and the risk of all-cause mortality in the middle-aged and elderly (≥45 years old) population in China. Methods Using data from five surveys from 2011-2020, years of education, age, gender, marital status, type of household, alcohol consumption status, smoking status, physical activity, limited ability to perform activities of daily living (ADLs), chronic disease status, and body mass index were collected. According to the survival status，the study participants were divided into a survival group (n=8625) and an all-cause mortality group (n=1735). Cox proportional risk regression model was used to analyze the relationship between years of education and the risk of all-cause mortality in middle-aged and elderly people with stratified analysis. Results The survey was conducted on 10360 research subjects, including 4 983 males and 5 377 females, with an age M(Q_R) of 59(8) years old. A total of 88 187 person years (average 8.512 person years) were followed up, and a total of 1735 deaths were reported over 9 years. The all-cause mortality rate was 19.674 ‰, with an education period M(Q_R) of 5(8) years and a survival time M(Q_R) of 9 (0) years. After adjusting for confounding factors, for every 1 year increase in education, the risk of all-cause mortality decreased by 2.60% [HR=0.974, 95% CI (0.960-0.988)]. The stratified analysis results showed that in the population aged 45-59, for every 1 year increase in education, the risk of all-cause mortality decreased by 1.00% [HR=0.990, 95% CI (0.959,1.021)]; In the population aged 60-74 and over 75, for every 1 year increase in education, the risk of all-cause mortality decreased by 2.70% [HR=0.973, 95% CI (0.948, 0.999)] and 3.50% [HR=0.965, 95% CI (0.929, 1.003)], respectively. Conclusion Improving the education level of middle-aged and elderly people (≥ 45 years old) in China can reduce the risk of all-cause mortality, and elderly education should be vigorously promoted.

Acute respiratory distress syndrome(ARDS)is a common respiratory emergency,but current clinical treatment remains at the level of symptomatic support and there is a lack of effective targeted treatment measures.Our previous study confirmed that inhalation of hydrogen gas can reduce the acute lung injury of ARDS,but the application of hydrogen has flammable and explosive safety concerns.Drinking hydrogen-rich liquid or inhaling hydrogen gas has been shown to play an important role in scavenging reactive oxygen species and maintaining mitochondrial quality control balance,thus improving ARDS in patients and animal models.Coral calcium hydrogenation(CCH)is a new solid molecular hydrogen carrier prepared from coral calcium(CC).Whether and how CCH affects acute lung injury in ARDS re-mains unstudied.In this study,we observed the therapeutic effect of CCH on lipopolysaccharide(LPS)induced acute lung injury in ARDS mice.The survival rate of mice treated with CCH and hydrogen inhalation was found to be comparable,demonstrating a significant improvement compared to the untreated ARDS model group.CCH treatment significantly reduced pulmonary hemorrhage and edema,and improved pulmonary function and local microcirculation in ARDS mice.CCH promoted mitochon-drial peripheral division in the early course of ARDS by activating mitochondrial thioredoxin 2(Trx2),improved lung mitochondrial dysfunction induced by LPS,and reduced oxidative stress damage.The results indicate that CCH is a highly efficient hydrogen-rich agent that can attenuate acute lung injury of ARDS by improving the mitochondrial function through Trx2 activation.

Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections.Herein,we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering(SERS)and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease(MNase)in serum samples.The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine(TMB)molecules to SERS-enhanced oxidized TMB(oxTMB),accompanied by the color change from colorless to blue.In the presence of S.aureus,the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads(MBs)to release alkaline phosphatase(ALP),which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away.Using this"on-to-off"triggering strategy,the target S.aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode.Meanwhile,the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis(n=7)and healthy participants(n=3),as well as monitored the prog-nostic progression of the disease(n=2).Overall,benefiting from highly active and dense"hot spot"substrate,MNase-mediated cascade response strategy,and colorimetric/SERS dual-signal output,this methodology will offer a promising avenue for the early diagnosis of S.aureus infection.

SUMMARY Hip fractures are common in elderly patients and are associated with significant morbidity and mortality,often referred to as the"last fracture of life".These fractures frequently result in a loss of functional independence.Evidence suggests that early surgical intervention can reduce mortality.The selection of treatment modality should take into account factors such as the type of fracture,the patient's age,and overall health status.This case report discusses an 88-year-old female patient who sustained an unstable intertrochanteric fracture of the left femur following a fall.She underwent closed reduction and internal fixation using an InterTAN intramedullary nail,resulting in a satisfactory postoperative recovery.Sixteen months following the surgical procedure,the patient presented with progressive pain in the left hip and ambulatory difficulties,absent from any evident trauma.Radiographic analysis identified a fracture of the left femoral neck accompanied by some degree of acetabular bone degradation attributable to the im-plant.Subsequently,the patient underwent removal of the internal fixation device and received a hemiar-throplasty.The postoperative course was uneventful,with marked improvements in both pain levels and functional capacity.This case underscored the intricate nature of femoral neck fractures following the internal fixation of intertrochanteric fractures.Contributing factors may include advanced age,osteoporo-sis,and stress shielding induced by the implant.In patients presenting with hip pain or gait disturbances months to years post-intertrochanteric fracture surgery,the potential for a new fracture should be consi-dered,even in the absence of an explicit traumatic incident.Radiographic imaging is imperative to exclude the presence of a fracture,particularly in individuals with high-risk factors such as advanced age,osteo-porosis,alcohol abuse,and a history of hormone therapy.Management of such cases may necessitate the removal of internal fixation devices and the implementation of hemiarthroplasty or total hip arthroplasty,contingent upon the patient's surgical tolerance.Crucially,anti-osteoporosis therapy serves as a vital pre-ventive strategy.Considering the high-risk profile of elderly patients with hip fractures,diligent follow-up and timely intervention are paramount to mitigating complications and mortality,thereby enhancing the quality of life for these patients.This case highlights the critical need for increased vigilance and compre-hensive management of elderly patients with hip fractures to enhance treatment outcomes and improve prognosis.

Objective:To construct a biosafety evaluation index system for major emergency public health events in medical institutions.Methods:Based on previous laboratory biosafety evaluation work, relevant regulations and standards on biosafety in China were collected through literature research and expert consultations. Candidate indicators for constructing the biosafety evaluation system for major emergency public health events in medical institutions were selected, and a framework was established. Two rounds of expert questionnaires were conducted to determine the content of the index system based on experts′ evaluation, and each indicator′s relevance and importance were scored. Finally, two rounds of Delphi consultations were carried out, and the Analytic Hierarchy Process (AHP) was applied to calculate the weights of indicators.Results:The response rates for the total four rounds of questionnaire surveys were all 100%. The first two rounds focused on determining the framework, while the latter two focused on determining the weights for each indicator. The authority coefficients of the expert consultations for the two rounds of weights were 0.65 and 0.70, respectively, indicating the reliability of the research results. In the final round of survey, the Kendall′s coefficients of concordance at each level were all greater than 0.1. Through statistical testing, the P-values were all less than 0.05, indicating good coordination of expert opinions. Ultimately, we established an operational biosafety evaluation system for major emergency public health events in medical institutions, consisting of 4 primary indicators, 26 secondary indicators, and 119 tertiary indicators, with additional deduction items, bonus items, unacceptable items, and monitoring indicators.Conclusions:Based on scientific theory, a biosafety evaluation system for major emergency public health events in medical institutions was constructed, achieving the integration of routine and emergency measures. This system can be used for self-assessment of laboratory biosafety during emergency public health events, addressing the lack of unified standards in biosafety evaluation. Through regular self-assessment, it can enhance the level of biosafety management in medical institution laboratories, to realize the value of application and dissemination.

Objective:To detect the expression levels of long non-coding RNAs(lncRNA)in elderly patients with pulmonary tuberculosis(PTB)and those with non-tuberculous lung diseases(non-TB), and to assess the performance of these lncRNA in the differential diagnosis of PTB.Methods:A total of 300 elderly patients with suspected PTB were recruited from Beijing Chest Hospital between January 2024 and September 2024, and were further divided into the PTB group and the non-TB lung disease group based on the results of mycobacterium tuberculosis(MTB)pathogenicity testing.Peripheral blood mononuclear cells were isolated using a lymphocyte separation solution, and RNA was extracted using the TRIzol method.Nine lncRNAs, previously identified as differentially expressed in PTB through our group's microarray analysis, were selected and detected by real-time fluorescence quantitative polymerase chain reaction to evaluate the expression levels of these lncRNAs between the PTB and non-TB lung disease groups.The overall patients were randomly divided into training and validation sets in a 7∶3 ratio.Lasso regression was employed to select the characteristic variables, and a random forest algorithm was then used to construct the lncRNA diagnostic portfolio.Receiver operating characteristic(ROC)curves were generated to evaluate the diagnostic performance of individual lncRNAs and the combined panel in differentiating elderly patients with PTB from those with other non-TB lung diseases.Results:A total of 201 cases were included, with 105 confirmed elderly patients diagnosed with PTB(52.2%)and 96 elderly patients suffering from non-TB lung disease(47.8%).Compared to the elderly patients with non-TB lung disease, the expression levels of ENST00000417346.1, ENST00000620744.1, lncRNA PWP1, ENST00000583184.1, lncRNA ABHD17B, ENST00000607464.1, ENST00000516057.1, and NR_003000 were significantly downregulated in the PTB patients, whereas the expression level of lncRNA BCL2L10 was significantly upregulated in the PTB patients.ROC analysis revealed that the area under the curve(AUC)for each lncRNA ranged from 0.659 to 0.848.The diagnostic panel, which included NR_003000, ENST00000607464.1, ENST00000583184.1, and ENST00000620744.1 as determined by Lasso analysis, exhibited AUC values of 0.917 and 0.906 in the training and validation sets, respectively.The performance of this panel was superior to that of each individual lncRNA.Conclusions:The random forest model, which incorporates NR_003000, ENST00000607464.1, ENST00000583184.1, and ENST00000620744.1, demonstrates potential in differentiating between PTB and non-TB lung diseases.

Objective To explore the therapeutic effect and potential mechanism of CA330 and OXIRIS adsorbent columns in septic shock.Methods Patients who met the diagnostic criteria for septic shock and admitted to the De-partment of Critical Care Medicine of Shenzhen Third People's Hospital from February 2022 to June 2024 were se-lected.They were randomly divided into an OXIRIS group and a CA330 group according to the random number table method.The CA330 group received hemoperfusion combined with hemodiafiltration using CA330 adsorbent co-lumn,while the OXIRIS group was treated with OXIRIS adsorbent column.Relevant markers of the two groups of patients before and after treatment were collected and compared,including inflammatory markers,bilirubin(total bilirubin[TBil],direct bilirubin[DBil]),coagulation functions(prothrombin time[PT],activated partial throm-boplastin time[APTT],etc),endotoxin(ETX),organ function scores(acute physiology and chronic health score Ⅱ[APACHE Ⅱ],sequential organ failure assessment[SOFA],etc).Molecular biology techniques were adopted to detect changes in inflammation-related gene expression(nuclear factor kappa B[NF-κB],toll-like receptor 4[TLR4],myeloid differentiation factor 88[MyD88]),and oxidative stress factors(glutathione peroxidase[GSH-Px],superoxide dismutase[SOD])in the blood of patients before and after treatment.The safety and effectiveness of two types of adsorbent columns during the treatment process was evaluated.Results A total of 92 patients were included and randomly divided into the OXIRIS group and the CA330 group,with 46 cases in each group.After treatment,the levels of TBil,DBil,and ETX in two groups of patients all showed significant decreases compared with before treatment(all P＜0.01),the levels of TBil,DBil,and ETX in patients in the OXIRIS group after treat-ment were all lower than those in the CA330 group during the same period(all P＜0.05);PT and APTT in both groups shortened significantly compared with before treatment(both P＜0.01),PT and APTT in the OXIRIS group after treatment were both shorter than those in the CA330 group during the same period(both P＜0.05);The APACHE Ⅱ score and SOFA score in patients in the OXIRIS group after treatment were both lower than those in the CA330 group during the same period(both P＜0.05);The levels of serum high-sensitivity C-reactive protein(hs-CRP),tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-5,and IL-8 in patients in both groups showed significant decreases compared with before treatment(all P＜0.05),and the levels of these serum markers in the CA330 group after treatment were all lower than those in the OXIRIS group during the same period(all P＜0.05).The gene expression levels of NF-κB,TLR4,and MyD88 in patients in the CA330 group after treatment were all lower than those in the OXIRIS group during the same period(all P＜0.05);The levels of GSH-Px and SOD in pa-tients in the OXIRIS group after treatment were both higher than those in the CA330 group(both P＜0.01).No serious adverse event occurred in patients in the CA330 group and the OXIRIS group during the treatment process.Conclusion OXIRIS may be better in clearing bilirubin and endotoxin,improving coagulation function,protecting organ function,and regulating oxidative stress response in patients,while CA330 may be more prominent in clearing inflammatory markers and regulating inflammation-related gene expression in patients.

[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

The clinical antiprotozoal drug nitazoxanide has been demonstrated to improve the experimental diabetes mellitus, lipid metabolism disorders, atherosclerosis and inhibit inflammation. Since the pathogenesis of heart failure with preserved ejection (HFpEF) is multifactorial and closely associated with the aforementioned diseases, we aim to study the effect of nitazoxanide on high-fat diet (HFD) plus L-NAME (N ω-nitro-l-arginine methyl ester)-induced HFpEF and metabolic syndrome in mice. We found that oral nitazoxanide improved cardiac hypertrophy, cardiac fibrosis, cardiac diastolic dysfunction, increased blood pressure, impaired exercise tolerance, impaired glucose handling, serum lipid disorders, hepatic steatosis, increased weight of white adipose tissues and kidney fibrosis in HFD + L-NAME-treated mice. In the established HFD + L-NAME-induced HFpEF and metabolic syndrome mouse model, therapeutic treatment with nitazoxanide rescued HFD + L-NAME-induced pathological phenotypes as mentioned above. The in vitro experiments revealed that tizoxanide, the active metabolite of nitazoxanide, increased the basal mitochondria metabolism of cardiomyocytes, inhibited cardiomyocyte hypertrophy and collagen secretion from cardiac fibroblasts, and relaxed phenylephrine- and U46619-induced constriction of rat mesenteric arteries, indicating that the direct effect of tizoxanide might partly contribute to the protective effect of nitazoxanide against HFpEF in vivo. The present study suggests that nitazoxanide might be a potential drug for HFpEF and metabolic syndrome therapy.