Objective To develop a precise method for analyzing urinary peptides based on electromembrane extraction(EME)combined with liquid chromatography-tandem mass spectrometry(LC-MS/MS),and to evaluate its potential applicability in tumor biomarker screening.Methods A total of 15 disease-associated peptides were selected as the target analytes.A supported liquid membrane(SLM)composed of n-octanol containing 5%di(2-ethylhexyl)phosphate was employed,with the donor phase being a 1∶1 mixture of urine and 100 mmol/L formic acid and urine,and the acceptor phase being 20 mmol/L formic acid containing 50%dimethyl sulfoxide(DMSO).After EME at 40 V for 15 min,the acceptor phase solution was analyzed by LC-MS/MS.Subsequently,the method,EME combined with LC-MS/MS(EME-LC-MS/MS),was preliminarily validated utilizing urine samples from 12 healthy controls and 7 patients with urinary system tumors.Results All 15 peptides exhibited excellent linearity in the range of 0.1-100.0 ng/mL(r≥0.995),with the limits of detection(LODs)being 0.01-0.50 ng/mL and the limits of quantification(LOQs)being 0.03-1.50 ng/mL.The spiked recoveries ranged from 21.0%to 71.2%,with relative standard deviations(RSDs)of 0.8%-20.0%(n=3).Small-sample analysis of clinical specimens revealed that the concentration of bradykinin 1-5 in the urine were significantly higher in tumor patients(median:0.65 ng/mL)than that in healthy controls(median:0.37 ng/mL)(P<0.05),suggesting its potential as a specific biomarker for urinary system tumors.Conclusion The EME-LC-MS/MS method established in the study features simplicity,high efficiency,and high sensitivity,enabling precise determination of trace-level peptides in urine samples.Moreover,this approach provides a reliable methodological basis for disease biomarker screening and promotes the clinical application of electromembrane extraction.

Objective:To explore the early diagnostic value and related mechanism of Lnc-IL7R/Jumonji domain-containing protein 3(Jmjd3)/keratinocyte growth factor(KGF)axis in inflammatory injury of acute respiratory distress syndrome(ARDS).Methods:① RT-qPCR and Western blot were used to detect Lnc-IL7R,Jmjd3,KGF mRNA and protein levels,as well as trimethyl-ation of histone H3 at lysine 27(H3K27)me3 protein level in serum of ARDS patients;ELISA was used to detect IL-1β,IL-6,TNF-α levels in serum.② Mitochondrial damage-associated molecular patterns(MTDs)was used to stimulate HBE cells and create traumatic ARDS cell model.HBE cells were divided into control group,model group(MTDs group),overexpression Lnc-IL7R group(Lnc-IL7R group),overexpression control group(NC group),overexpression Lnc-IL7R+overexpression Jmjd3 group(Lnc-IL7R+Jm-jd3 group).Chromatin immunoprecipitation(ChIP)was used to detect Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in pro-moter region;qRT-PCR and Western blot were used to detect Lnc-IL7R,Jmjd3,KGF,IL-1β,IL-6,TNF-α mRNA and protein levels,as well as H3K27me3 protein level.③ Injection of MTDs into tail vein of mice was used to create traumatic ARDS mice model.Eighty mice were divided into control group,MTDs group,Lnc-IL7R group,NC group and Lnc-IL7R+Jmjd3 group,with sixteen mice in each group.HE staining was used to detect pathological damage in lung tissue;ChIP and RT-qPCR were used to detect Lnc-IL7R,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region and mRNA levels;ELISA was used to detect IL-1β,IL-6,TNF-α protein levels in BALF;Western blot was used to detect Jmjd3,KGF,H3K27me3 protein levels in lung tissue;immunohisto-chemical staining was used to detect surfactant protein C(SPC)and surfactant protein D(SPD)protein levels in lung tissue.Results:①Compared with healthy individuals,Lnc-IL7R,Jmjd3,KGF mRNA and protein levels,IL-1β,IL-6,TNF-α protein levels in serum of ARDS patients were significant increased,while H3K27me3 protein level was significant decreased(P<0.05).②Compared with control group,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,H3K27me3 protein level in HBE of MTDs group were significant decreased,while Lnc-IL7R,Jmjd3,KGF,IL-1β,IL-6,TNF-α mRNA and protein levels,Lnc-IL7R were significant increased(P<0.05);after overexpression of Lnc-IL7R,Lnc-IL7R,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,KGF mRNA and protein levels,H3K27me3 protein level were significant increased,Jmjd3,IL-1β,IL-6,TNF-α mRNA and protein levels in lung tissue and BALF were significant decreased(P<0.05);overexpression of Jmjd3 could partially reverse the effects of overexpression of Lnc-IL7R on the above indicators(P<0.05).③Compared with Control group,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,H3K27me3 protein level in lung tissue of MTDs group were significant decreased,while Lnc-IL7R,Jmjd3,KGF,IL-1β,IL-6,TNF-α mRNA and protein levels,H3K27me3,SPC,SPD protein levels in lung tissue and BALF were significant increased(P<0.05);after overexpression of Lnc-IL7R,Lnc-IL7R,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,KGF mRNA and protein levels,H3K27me3,SPC,SPD protein levels in lung tissue were significant increased,while Jmjd3,IL-1β,IL-6,TNF-α mRNA and protein levels in lung tissue and BALF were significant decreased(P<0.05);overexpression of Jmjd3 could partially reverse the effects of overexpression of Lnc-IL7R on the above indicators(P<0.05).Conclusion:Lnc-IL7R/Jmjd3/KGF axis participates in inflammation regulation and alveolar epithelial cell injury repair in ARDS by regulating histone methylation modifications in the promoter region.Lnc-IL7R/Jmjd3/KGF can be used for early diagnosis of ARDS.

Objective:To explore the early diagnostic value and related mechanism of Lnc-IL7R/Jumonji domain-containing protein 3(Jmjd3)/keratinocyte growth factor(KGF)axis in inflammatory injury of acute respiratory distress syndrome(ARDS).Methods:① RT-qPCR and Western blot were used to detect Lnc-IL7R,Jmjd3,KGF mRNA and protein levels,as well as trimethyl-ation of histone H3 at lysine 27(H3K27)me3 protein level in serum of ARDS patients;ELISA was used to detect IL-1β,IL-6,TNF-α levels in serum.② Mitochondrial damage-associated molecular patterns(MTDs)was used to stimulate HBE cells and create traumatic ARDS cell model.HBE cells were divided into control group,model group(MTDs group),overexpression Lnc-IL7R group(Lnc-IL7R group),overexpression control group(NC group),overexpression Lnc-IL7R+overexpression Jmjd3 group(Lnc-IL7R+Jm-jd3 group).Chromatin immunoprecipitation(ChIP)was used to detect Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in pro-moter region;qRT-PCR and Western blot were used to detect Lnc-IL7R,Jmjd3,KGF,IL-1β,IL-6,TNF-α mRNA and protein levels,as well as H3K27me3 protein level.③ Injection of MTDs into tail vein of mice was used to create traumatic ARDS mice model.Eighty mice were divided into control group,MTDs group,Lnc-IL7R group,NC group and Lnc-IL7R+Jmjd3 group,with sixteen mice in each group.HE staining was used to detect pathological damage in lung tissue;ChIP and RT-qPCR were used to detect Lnc-IL7R,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region and mRNA levels;ELISA was used to detect IL-1β,IL-6,TNF-α protein levels in BALF;Western blot was used to detect Jmjd3,KGF,H3K27me3 protein levels in lung tissue;immunohisto-chemical staining was used to detect surfactant protein C(SPC)and surfactant protein D(SPD)protein levels in lung tissue.Results:①Compared with healthy individuals,Lnc-IL7R,Jmjd3,KGF mRNA and protein levels,IL-1β,IL-6,TNF-α protein levels in serum of ARDS patients were significant increased,while H3K27me3 protein level was significant decreased(P<0.05).②Compared with control group,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,H3K27me3 protein level in HBE of MTDs group were significant decreased,while Lnc-IL7R,Jmjd3,KGF,IL-1β,IL-6,TNF-α mRNA and protein levels,Lnc-IL7R were significant increased(P<0.05);after overexpression of Lnc-IL7R,Lnc-IL7R,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,KGF mRNA and protein levels,H3K27me3 protein level were significant increased,Jmjd3,IL-1β,IL-6,TNF-α mRNA and protein levels in lung tissue and BALF were significant decreased(P<0.05);overexpression of Jmjd3 could partially reverse the effects of overexpression of Lnc-IL7R on the above indicators(P<0.05).③Compared with Control group,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,H3K27me3 protein level in lung tissue of MTDs group were significant decreased,while Lnc-IL7R,Jmjd3,KGF,IL-1β,IL-6,TNF-α mRNA and protein levels,H3K27me3,SPC,SPD protein levels in lung tissue and BALF were significant increased(P<0.05);after overexpression of Lnc-IL7R,Lnc-IL7R,Jmjd3,IL-1β,IL-6,TNF-α histone methylation levels in the promoter region,KGF mRNA and protein levels,H3K27me3,SPC,SPD protein levels in lung tissue were significant increased,while Jmjd3,IL-1β,IL-6,TNF-α mRNA and protein levels in lung tissue and BALF were significant decreased(P<0.05);overexpression of Jmjd3 could partially reverse the effects of overexpression of Lnc-IL7R on the above indicators(P<0.05).Conclusion:Lnc-IL7R/Jmjd3/KGF axis participates in inflammation regulation and alveolar epithelial cell injury repair in ARDS by regulating histone methylation modifications in the promoter region.Lnc-IL7R/Jmjd3/KGF can be used for early diagnosis of ARDS.

Objective To investigate the effect of sarcopenia on balance function in patients with Parkinson's disease(PD).Methods Sixty patients with PD diagnosed by our hospital from January 2021 to June 2021 were selected as the study objects.All patients were divided into myopenia group and control group according to the diagnostic criteria established by the Asian Working Group for Sarcopenia(AWGS)in 2014.The UPDRS Ⅲ,Berg balance scale,timed-up-go test and activity balance confidence scale were compared between the two groups.Results The grip strength,walking speed and RASM of the myopenia group were lower than those of the control group(P<0.05);Compared with the control group,the number of falls,UPDRS Ⅲ,Berg score and TUG duration in the myopenia group were significantly increased(P<0.05).The total score of ABC,ABC-2,ABC-3,ABC-5,ABC-6,ABC-7,ABC-8,ABC-9,ABC-11,ABC-14,ABC-15,ABC-16 in myopenia group were poor,and the differences were statistically significant(P<0.05).Further analyzing the differences between sex genders,we found that female sarcopenia had lower scores on ABC-2,ABC-3,ABC-8,ABC-9,ABC-11,ABC-13,ABC-14,ABC-15,ABC-16,and total score indexes,with statistically significant differences.Conclusion Myopenia significantly affects the balance function of PD and increases the risk of falls.

Objective To investigate the value of serum aquaporin 1(AQP1)level combined with extra-vascular lung water index(EVLWI)in assessing the severity and prognosis of sepsis causing acute respiratory distress syndrome(ARDS).Methods 268 patients with sepsis-induced ARDS(ARDS group)and 55 patients with sepsis alone(sepsis alone group)admitted to our hospital from January 2020 to December 2023 were selected.Patients with sepsis-induced ARDS were divided into 89 mild,109 moderate,and 70 severe groups according to the oxygenation index(OI),and 104 deaths and 164 survivors according to the 28 d prognosis.Detection of serum AQP1 levels and calculation of EVLWI.Using Spearman's method,the correlation between serum AQP1 level,EVLWI and OI in patients with sepsis-induced ARDS;the establishment of a logistic regression model to determine the factors of sepsis-induced mortality in patients with ARDS;and the plotting of ROC curves to evaluate the value of serum AQP1 level in combination with EVLWI for its assessment.Results Compared with the simple sepsis group,serum AQP1 level was reduced and EVLWI was increased in the ARDS group(P<0.05).AQP1 levels were sequentially lower and EVLWI sequentially higher in the mild,moderate,and severe groups(P<0.05).Serum AQP1 levels were positively correlated with OI in patients with sepsis-induced ARDS,and EVLWI was negatively correlated with OI in patients with sepsis-induced ARDS(P<0.05).The 28 d mortality rate in 268 patients with ARDS due to sepsis was 38.81%(104/268).The independent protective factors for death in ARDS patients with sepsis were elevated OI(OR=0.984,95%CI:0.976～0.992)and elevated AQP1(OR=0.761,95%CI:0.677～0.854),and the independent risk factors were increased SOFA score(OR=1.367,95%CI:1.142～1.636)and elevated blood lactate(OR=2.515,95%CI:1.689～3.745),and elevated EVLWI(OR=1.559,95%CI:1.290～1.885)(P<0.05).The AUC predicted by serum AQP1 level combined with EVLWI was 0.887(95%CI:0.843～0.923),which was greater than the AUC predicted by serum AQP1 level,and EVLWI alone,which was 0.792(95%CI:0.738～0.839),and 0.807(95%CI:0.754～0.852)(P<0.05).Conclusion Decreased serum AQP1 levels and elevated EVLWI were associated with increased severity and poor prognosis in patients with sepsis-induced ARDS,and serum AQP1 levels in combination with EVLWI were of high value in assessing the prognosis of patients with sepsis-induced ARDS.

Humans ; Hypertension, Pulmonary/surgery* ; Learning Curve ; Angioplasty, Balloon ; Pulmonary Embolism/therapy* ; Chronic Disease ; Pulmonary Artery/surgery*

Objective:To investigate the clinical efficacy and safety of intravenous thrombolysis with different doses of alteplase in the treatment of acute cerebral infarction in older adult patients.Methods:A total of 65 older adult patients with acute cerebral infarction (onset within 4.5 hours, age ≥ 75 years) who underwent intravenous thrombolysis in Wenzhou Central Hospital from February 2021 to February 2022 were included in this study. They were randomly assigned to undergo intravenous thrombolysis with either low dose alteplase (0.6 mg/kg, low dose group, n = 32) or standard dose alteplase (0.9 mg/kg, standard dose group, n = 33). The National Institutes of Health Neurological Stroke Scale score before and 24 and 48 hours after treatment, modified Rankin scale score before and 7, 14 and 90 days after treatment, serum C-reactive protein (CRP), neuron-specific enolase (NSE) and tumor necrosis factor-α (TNF-α) levels before and 24 hours after treatment, 24-hour incidence of intracranial hemorrhage, 24-hour incidence of symptomatic intracranial hemorrhage, and 90-day mortality were compared between the two groups. Results:Compared with before treatment, the National Institutes of Health Neurological Stroke Scale scores in each group were significantly decreased at 24 and 48 hours after treatment (low dose group, t24 h = 6.78, t48 h = 7.86; standard dose group: t24 h = 8.09, t48 h = 10.13, all P < 0.001). Compared with before treatment, the modified Rankin scale score in each group was significantly decreased at 7, 14 and 90 days after treatment (low-dose group: t7 d = 5.19, t14 d = 8.47, t90 d = 9.85; standard dose group: t7 d = 6.83, t14 d = 7.74, t90 d = 13.66, all P < 0.001). At 24 hours after treatment, serum levels of CRP, NSE, TNF-α in each group were significantly decreased (low-dose group: tCRP = 5.13 , tNSE = 4.22, tTNF-α = 34.29; standard dose group: tCRP = 4.87, tNSE = 5.53, tTNF-α = 31.98, all P < 0.001). At each time point after treatment, there were no significant differences in these indices between the two groups (all P > 0.05). The 24-hour incidence of intracranial hemorrhage in the low dose group was significantly lower than that in the standard dose group ( χ2 = 4.58, P = 0.032). There were no significant differences in incidence of symptomatic intracranial hemorrhage and 90-day mortality between the two groups (all P > 0.05). Conclusion:Intravenous thrombolysis with low dose alteplase (0.6 mg/kg) for the treatment of acute cerebral infarction in older adult patients exhibits equivalent clinical efficacy to that with standard dose alteplase (0.9 mg/kg), and the former is much safer than the latter.

Atopic dermatitis (AD) is a chronic, relapsable and pruritic skin disease, commonly found in children and adolescents. The prevalence of AD is increasing worldwide. It is reported that AD is related to many factors such as genetic inheritance, environment, immunity and skin barrier dysfunction, suggesting a very complex pathogenesis. In recent years, high-throughput technologies in the field of genomics and metabolomics have opened up new perspectives on the pathogenesis of AD, and shown potential application prospects in microbiome transplantation therapies for AD. This review summarized the current advances in the relationship between skin microecology and skin health, the pathogenesis and microbiomic characteristics of AD, features of pathogenic microorganisms, and microbiome transplantation therapies for AD. Based on our own practical experience, we put forward a culturomics research protocol to study the human skin microbiome and a method for quantitative microbiological examination, aiming to provide reference for the prevention, clinical treatment and therapeutic monitoring of AD.

Modern medical imaging techniques,such as computed tomorgraphy (CT),magnetic resonance imaging (MRI) and position emission tomorgraphy/computed tomorgraphy (PET-CT) can accurately delineate the gross target volume (GTV) of hepatocellular carcinoma (HCC).Comparison of postoperative pathological subclinical lesions,imaging and clinical parameters contributes to the precise delineation of clinical target volume (CTV).Moreover,radiotherapy-assisted techniques,such as fourdimensional computed tomography (4DCT),compression of abdomen,active breathing control and respiratory gating,can minimize the internal target volume (ITV).In addition,immobilization with vacuum cushion and body membrane can reduce the set-up error,minimize the planning target volume (PTV) and avoid or decrease the irradiation error or missing irradiation.All these approach can minimize the target volume,elevate the dose and reduce the complications during radiotherapy for HCC.In this article,the research progress on the target delineation for external beam radiotherapy in HCC patients was reviewed.

Surgical resection or liver transplantation are the primary curative treatments for hepatocellular carcinoma(HCC).However, a minority of patients are eligible for these treatment strategies upon initial diagnosis.With the development of radiotherapy technol-ogy,radiotherapy is being used increasingly in the treatment of HCC.A large number of studies have shown that external beam radio-therapy has obvious advantages for different stages of HCC.Stereotactic radiotherapy provides an alternative to radiofrequency abla-tion in the treatment of early unresectable HCC,and radiotherapy combined with other local or systemic treatments,including trans-catheter arterial chemoembolization and sorafenib,has been applied for the treatment of locally advanced and distant metastatic le-sions.The particle beam has the advantages of Bragg peak and better dose distribution compared to conventional radiography,show-ing promising prospect in clinical application.This review focuses on the developments in the field of external beam radiotherapy in the treatment of HCC.