Objective To establish a stable liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for detecting gamma-aminobutyric acid(GABA)in plasma,and to evaluate the value of GABA detection in the diagnosis of sleep disorders.Methods GABA was detected using a UPLC Xevo TQs system.The method was pre-validated and its performance was verified to establish a reference range for healthy individuals.The difference in plasma GABA levels between apparently healthy individuals and patients with sleep disorders was compared.Results We employed deuterated compounds as isotopic internal standards and utilized an Amide chromatographic column for separation.The mobile phase was 0.050%formic acid in water and 90%acetonitrile in water containing 0.175%formic acid and 5 mmol/L ammonium acetate with gradient elution in the column temperature of 35℃.The linear range for the detection of GABA by LC-MS/MS was 0.05-10.00 μmol/L,with a lower limit of quantification of 0.02 μmol/L,the inter-day CV<3.00%and intra assay CV<4.00%,respectively,and the recovery rate was 101.06%-109.02%.The reference ranges for plasma GABA were established by analyzing 300 healthy controls stratified by age:18-34 years(0.08-0.15 μmol/L),35-49 years(0.10-0.20 μmol/L),and≥50 years(0.12-0.23 μmol/L).Then plasma GABA was used as a biomarker for auxiliary diagnosis of sleep disorders in analyzing 221 patients and 300 healthy controls,which revealed that AUC values were 0.510(P=0.850),0.686(P=0.002),and 0.890(P<0.001)in the groups of 18-34 years,35-49 years,and≥50 years,respectively,with optimal cut-off values of 0.09,0.10 and 0.11 μmol/L.Conclusion A reliable LC-MS/MS method for detecting GABA has been established,which can detect plasma GABA levels sensitively and accurately and can be used in assisting the clinical diagnosis of sleep disorders.

Purpose To investigate the change of network topology attribute of brain structure in obese adolescents using graph theory analysis.Materials and Methods A total of 86 obese adolescents in Children's Hospital of Nanjing Medical University from January 2023 to April 2024 were prospectively collected,including 60 patients in the simple obesity group(OB group),26 patients in the obese group with metabolic syndrome(MS group).Meanwhile,24 healthy volunteers were recruited as the control group.Diffusion tensor imaging scan of the head,anthropological characteristics examination and blood test were performed.The differences of global and local topology properties among the three groups were compared.Global or node attributes with statistical differences were correlated with clinical indicators.Results There were significant differences in body height,body weight,body mass index,waist circumference,diastolic blood pressure,systolic blood pressure and high density lipoprotein cholesterol among the three groups(χ2/F=3.208-5.085,all P<0.05).Compared with the control group,there were significant differences in total body fat,triglyceride,total cholesterol,alanine amino-transferase,aspartate aminotransferase and fasting insulin between the OB group and the MS group(all P<0.05).No statistically significant differences were observed in the small world attribute,shortest path length and global efficiency among the three groups(all P>0.05).A statistically significant difference was observed in the clustering coefficient and local efficiency among the three groups(F=2.534,3.012,both P<0.05),with the values of clustering coefficient and local efficiency of OB group were significantly lower than those of the control group(P<0.05).The node attribute differences occurred in the left frontal lobe,right insula,right anterior central gyrus,left anterior cingulate gyrus,left anterior cuneus and bilateral occipital lobes.The connection value between module Association and Limbic in control group was significantly lower smaller than those of the OB group(P=0.033).A negative correlation was observed between local efficiency and age(r=-0.239),body weight(r=-0.254),body mass index(r=-0.230)and waist circumference(r=-0.263,all P<0.05).The modularity was negatively correlated with fasting insulin(r=-0.325,P<0.05).Conclusion Adolescent obesity leads to the abnormal topological properties of structural network in some brain regions,which may be associated with the abnormal cognitive function and could provide neuroimaging evidence for clinical intervention.

Objective The pathophysiological process of acute high-altitude hypoxia-induced lung injury affects protein expression levels,which are mainly evaluated by Western blot.No systematic study has investigated changes in internal control proteins as calibration loading amounts.Methods Lung injury at an altitude of 6000 m was induced in a low-pressure,low-oxygen chamber for 8,24,and 72 h using C57BL/6J mice.Establishment of the model was confirmed by hematoxylin and eosin staining.Expression levels of various internal control proteins,including vinculin,α-tubulin,eukaryotic translation initiation factor 5(EIF5),β-actin,and glyceraldehyde 3-phosphate dehydrogenase(GAPDH)were detected by Western blot,and total protein expression was detected by Coomassie blue staining.Furthermore,the lung injury model in vitro was establised by using,Bronchial epithelial cell(BZAS-2B)andhunman umbilical vein endothelial cells(HUVECS)confirmed by TUNEL staining.Expression levels of internal control proteins were detected by Western blot,and total protein expression was detected by Coomassie Blue staining.Results Acute 8,24,and 72 h hypoxic models were successfully established in lung tissue,demonstrating consistent total protein expression and stable levels of the internal reference proteins vinculin,α-tubulin,EIF5,andβ-actin.GAPDH expression was elevated in the HH8 h,HH24 h,and HH72 h groups compared with the normoxia(Nor)group,but only the increase at HH72 h groups was significant.Similarly,8,24,and 48 h hypoxic models were successfully established in BEAS-2B cells and HUVECs,with consistent total protein expression.In BEAS-2B cells,expression levels of the internal reference proteins β-actin and GAPDH were consistent with the normoxic control(NC)group,while vinculin,α-tubulin,and EIF5 expression levels were significantly reduced under hypoxic conditions for up to 24 h.In HUVECs,vinculin and α-tubulin expression levels were also consistent with the NC group,while EIF5,β-actin,and GAPDH expression levels were significantly reduced at 8 h and increased at 48 h.Conclusions Acute hypoxia induces lung tissue injury,and protein expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,and β-actin are stable,making them suitable internal references for Western blot.Additionally,Western blot detected differential expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,β-actin,and GAPDH in BEAS-2B cells and HUVECs,as the most important in vitro lung tissue models of hypoxia-induced injury.

Objective The pathophysiological process of acute high-altitude hypoxia-induced lung injury affects protein expression levels,which are mainly evaluated by Western blot.No systematic study has investigated changes in internal control proteins as calibration loading amounts.Methods Lung injury at an altitude of 6000 m was induced in a low-pressure,low-oxygen chamber for 8,24,and 72 h using C57BL/6J mice.Establishment of the model was confirmed by hematoxylin and eosin staining.Expression levels of various internal control proteins,including vinculin,α-tubulin,eukaryotic translation initiation factor 5(EIF5),β-actin,and glyceraldehyde 3-phosphate dehydrogenase(GAPDH)were detected by Western blot,and total protein expression was detected by Coomassie blue staining.Furthermore,the lung injury model in vitro was establised by using,Bronchial epithelial cell(BZAS-2B)andhunman umbilical vein endothelial cells(HUVECS)confirmed by TUNEL staining.Expression levels of internal control proteins were detected by Western blot,and total protein expression was detected by Coomassie Blue staining.Results Acute 8,24,and 72 h hypoxic models were successfully established in lung tissue,demonstrating consistent total protein expression and stable levels of the internal reference proteins vinculin,α-tubulin,EIF5,andβ-actin.GAPDH expression was elevated in the HH8 h,HH24 h,and HH72 h groups compared with the normoxia(Nor)group,but only the increase at HH72 h groups was significant.Similarly,8,24,and 48 h hypoxic models were successfully established in BEAS-2B cells and HUVECs,with consistent total protein expression.In BEAS-2B cells,expression levels of the internal reference proteins β-actin and GAPDH were consistent with the normoxic control(NC)group,while vinculin,α-tubulin,and EIF5 expression levels were significantly reduced under hypoxic conditions for up to 24 h.In HUVECs,vinculin and α-tubulin expression levels were also consistent with the NC group,while EIF5,β-actin,and GAPDH expression levels were significantly reduced at 8 h and increased at 48 h.Conclusions Acute hypoxia induces lung tissue injury,and protein expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,and β-actin are stable,making them suitable internal references for Western blot.Additionally,Western blot detected differential expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,β-actin,and GAPDH in BEAS-2B cells and HUVECs,as the most important in vitro lung tissue models of hypoxia-induced injury.

OBJECTIVE To explore the clinical characteristics of the nontuberculous Mycobacteria pulmonary dis-ease(NTMPD)patients with previous pulmonary tuberculosis(PPTB)and analyze the clinical difference from the recurrence of pulmonary tuberculosis.METHODS By means of retrospective survey,the patients who were diag-nosed with NTMPD and recurrent pulmonary tuberculosis in Wuhan Pulmonary Hospital from Mar.2021 to Oct.2023 were recruited as the research subjects,a total of 395 patients with NTMPD were enrolled in the study and were divided into the PPTB-NTMPD group with 92 cases and the NPPTB-NTMPD group with 303 cases according to the history of PPTB.The baseline data,clinical symptoms,imaging findings,underlying diseases,pulmonary diseases,and species of nontuberculous Mycobacteria(NTM)were observed and compared.Totally 92 patients with recurrent pulmonary tuberculosis were randomly screened and assigned as the recurrent pulmonary tuberculo-sis group in a 1:1 ratio by matching the PPTB-NTMPD group with the gender and age.The major clinical charac-teristics were compared between the two groups.The 92 patients with PPTB-NTMPD were divided into the 1-10 years group with 40 cases,the 10-30 years group with 37 cases,and the more than 30 years group with 15 cases according to the interval between the initial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD.The major clinical characteristics were compared among the groups.RESULTS The age was(64.21±10.71)years old in the PPTB-NTMPD group,(60.26±11.83)years old in the NPPTB-NTMPD group(t=3.020,P=0.003).The proportion of patients with body mass index less than 18.5 kg/m2 was 59.78％in the PPTB-NTMPD group,41.25％in the NPPTB-NTMPD group(x2=6.155,P=0.013);the proportion of patients with cough was 77.17％in the PPTB-NTMPD group,65.68％in the NPPTB-NTMPD group(x2=4.313,P=0.038);the inci-dence of cavitary shadow was 50.00％in the PPTB-NTMPD group,35.31％in the NPPTB-NTMPD group(x2=6.414,P=0.011);the incidence of emphysema and pulmonary bullae was 29.35％in the PPTB-NTMPD group,12.87％in the NPPTB-NTMPD group(x2=13.766,P＜0.001);the incidence of chronic obstructive pulmonary disease(COPD)was 22.83％in the PPTB-NTMPD group,14.19％in the NPPTB-NTMPD group(x2=3.875,P=0.049);the incidence of damaged lung was 9.78％in the PPTB-NTMPD group,2.97％in the NPPTB-NT-MPD group(x2=7.530,P=0.014);there were significant differences.Mycobacterium intracellulare and Myco-bacterium abscessus were the predominant species of NTM in both the PPTB-NTMPD group and the NPPTB-NT-MPD group,there was no significant difference in the distribution of NTM species between the two groups of pa-tients.The incidence of patch shadow of the PPTB-NTMPD group was lower than that of the recurrent pulmonary tuberculosis group(P＜0.05),the incidence of bronchiectatic shadow of the PPTB-NTMPD group was higher than that of the recurrent pulmonary tuberculosis group(P＜0.05).There were significant differences in the age,incidence of pleural thickening and incidence of COPD among the patients with different time intervals between ini-tial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD in the PPTB-NTMPD group(P＜0.05).CONCLUSIONS The previous pulmonary tuberculosis mainly affect the body mass index less than 18.5 kg/m2 and the post-tuberculosis pulmonary diseases such as cough,pulmonary cavity,emphysema,pulmonary bullae,COPD and damaged lung of the NTMPD patients.The NTMPD patients with previous pulmonary tuberculosis are more likely to have bronchiectasia than the patients with recurrent tuberculosis.It is necessary for the clinicians to attach great importance.

Aim To identify the underlying target of bullatine A(BA)against rheumatoid arthritis(RA)u-sing limited proteolysis-small molecule mapping(LiP-SMap)drug target proteomics and to provide a scientif-ic basis for clinical application of Aconiti brachypodi Radix in the treatment of RA.Methods LiP-SMap drug target proteomics was employed to perform bioin-formatics analysis for comparing and validating the dif-ferential protein expression after BA intervention.A collagen-induced arthritis(CIA)model was estab-lished in DBA/1 mice using bovine type Ⅱ collagen.The mice were then divided into the CIA model group,methotrexate-positive control group(MTX group),and BA groups(10 mg·kg-1 and 20 mg·kg-1)based on their clinical scores.After drug intervention,the thera-peutic efficacy against RA was assessed by joint index scores and foot thickness measurements.Histopatholog-ical changes in the arthritic joints of CIA mice were e-valuated using hematoxylin and eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was employed to detect inflammatory cytokines interleukin-17(IL-17)and total IgG and IgG3 anti-collagen-spe-cific antibodies levels from the serum of CIA mice.Flow cytometry was used to detect the expression levels of intracellular Th17 cells(IL-17+CD4+T cells)and Th1 cells(IFN-γ+CD4+T cells).Fluorescent quanti-tative PCR was performed to detect the expression of genes related to differential proteins.Results The proteomic analysis identified Serpinb1a as a protein with strong binding affinity to BA,and KEGG enrich-ment analysis indicated IL-17 signaling pathway was a crucial pathway of BA in against RA.BA treatment significantly reduced clinical scores and foot thickness,improved local arthritis symptoms in CIA mice,and al-leviated inflammatory cell infiltration into arthritic joints(P＜0.05).Differential protein validation re-sults showed that BA had strong affinity with Serpinb1a(-5.92 kJ·mol-1)and downregulated the expres-sion of Serpinb1a mRNA.Furthermore,the administra-tion of BA markedly reduced serum IL-17 A levels from CIA mice,inhibited the expression of intracellular IL-17 A and IFN-γ cytokines in splenic CD4+T cells(P＜0.05),and significantly downregulated the transcrip-tional expression of IL-17F(P＜0.05).Conclusion BA exhibits therapeutic effects on collagen-induced arthritis,and its mechanism of action may involve the regulation of Serpinb1a and the IL-17 signaling path-way.

Objective:To explore the construction and efficacy of cranial magnetic resonance imaging(MRI)classification model based on attention mechanism in type 2 diabetes patients with cognitive impairment.Methods:The case data of 100 patients with type 2 diabetes who were treated in the General Hospital of North China Petroleum Administration Bureau from June 2022 to January 2024 were retrospectively selected.A total of 100 MRI images of cranial FLAIR_LongTR sequence with cognitive impairment(32 cases)and those without cognitive impairment(68 cases)were respectively collected.The images of the above two kinds of samples were horizontally and vertically translated to expand to 1000 samples,respectively.The samples were randomly divided into training samples(n=700)and test samples(n=300)as the ratio of 7:3 according to affine transformation data augmentation method.Then,the attention mechanism model was established to test the images with full scan of the test samples.The ability of the attention mechanism system in screening cognitive impairment was analyzed according to the method of setting threshold value.The 100 MRI images of cranial FLAIR_LongTR sequence of patients in our hospital from January 2024.From January to May 2024 were used as a verification set to verify the diagnostic value of attention mechanism.Results:With the increasing of iteration times,the sample loss of training and verification of attention mechanism model gradually decreased and tended toward stability,and the accuracy of training set and verification set gradually increased and tended toward stability.In the attention mechanism model,the average loss rate of training samples was 10.024%,and that of test samples was 15.247%.In the attention mechanism model,the average accuracy of training samples was 99.078%,and the average accuracy of test samples was 99.753%.Receiver operating characteristic(ROC)curves showed that the area under curve(AUC)of attention mechanism model was 0.998,which can better diagnose cognitive impairment of patients with type 2 diabetes than the resNET model(AUC=0.656)(Z=3.437,P<0.001).Conclusion:The constructed cranial MRI classification model by using attention mechanism has favorable diagnostic value for cognitive impairment in patients with type 2 diabetes.

Objective:To investigate effect of quality improvement based on the Attitude-Skill-Knowledge(ASK)model on quality of life and emotion in patients with acute myocardial infarction(AMI).Methods:This random-ized controlled study enrolled 220 AMI patients admitted in Liyang People's Hospital between October 2020 and Oc-tober 2022.They were divided into control group(n=110,routine nursing)and intervention group(n=110,addi-tional quality improvement intervention based on ASK model).After 2-month intervention,scores of Patient Health Questionaire-9(PHQ-9),Beck Anxiety Inventory(BAI)and General Quality of life Inventory(GQOLI-74),cardiac function and satisfaction for nursing were compared between two groups.Results:After 2-month intervention,compared with patients in control group,those in intervention group had significant higher score of GQOLI-74[(372.82±23.07)points vs.(306.48±15.53)points],significant lower scores of PHQ-9[(9.58±2.41)points vs.(15.36±3.51)points]and BAI[(8.61±0.24)points vs.(16.81±3.54)points],levels of N-terminal pro brain natriuretic peptide(NT-proBNP)[(5.03±1.15)pg/ml vs.(13.11±1.22)pg/ml],cardiac troponin I(cTnⅠ)[(2.90±0.51)pg/ml vs.(4.76±1.32)pg/ml]and creatine kinase isoenzyme MB(CK-MB)[(8.70±2.31)ng/ml vs.(13.63±2.63)ng/ml](P＜0.001 all).Total satisfaction for nursing in intervention group was significantly higher than that of control group(94.55％vs.83.64％,P=0.009).Conclusion:Quality improvement based on Attitude-Skill-Knowledge model could improve cardiac function and quality of life as well as negative emotion in AMI patients with high satisfaction.

Objective:To translate the Short Self-Regulation Questionnaire (SSRQ) into Chinese, and test its reliability and validity in breast cancer patients.Methods:Using the Brislin translation model, cross-cultural adaptation was conducted by combining expert consultation and a pre-survey on SSRQ. From August to December 2023, 324 breast cancer patients from Department of Breast Surgery of Peking University First Hospital were selected by convenience sampling to test the reliability and validity of SSRQ.Results:A total of 324 questionnaires were distributed and 307 valid questionnaires were recovered, with an effective recovery rate of 94.75% (307/324). The final questionnaire consisted of 30 items. The content validity index at the item level ranged from 0.80 to 1.00, and the content validity index at the scale level was 0.97. The predictive validity result showed a good model fit. The Cronbach's α coefficient for the questionnaire was 0.90, the folded reliability coefficient was 0.85, and the retest reliability coefficient was 0.79.Conclusions:SSRQ has good reliability and validity in breast cancer patients and has value for clinical promotion.

Objective:To retrieve, summarize, evaluate, and synthesize the best available evidence on helicopter transport nursing care for critically ill patients in order to provide an evidence-based reference for clinical practice.Methods:A systematic search was conducted across guideline repositories, professional society websites, and both Chinese and international databases for all relevant evidence regarding helicopter transport nursing care for critically ill patients, covering the period from database inception to January 22, 2024. Three researchers independently conducted quality appraisals of the included literature, and two researchers extracted and synthesized the evidence.Results:A total of 13 articles were included, comprising five guidelines, seven expert consensus, and one national standard. Twenty-nine pieces of evidence were summarized, covering four aspects: assessment and decision-making, pre-transfer preparation, en route care, and patient handover.Conclusions:This study synthesizes the best available evidence on helicopter transport nursing care for critically ill patients and provides an evidence-based foundation for the implementation of helicopter-based patient transfer nursing practices.