Objective To establish a fluorescent recombinase-aided amplification (RAA) assay for detection of Strongyloides stercoralis nucleic acid and to preliminarily evaluate its performance. Methods Six sets of specific primers targeting S. stercoralis 18S ribosomal RNA (18S rRNA) gene and one fluorescent probe were designed and synthesized. The optimal primer-probe set was determined through systematic screening and optimization to establish the fluorescent RAA assay. The assay was evaluated using S. stercoralis genomic DNA at concentrations of 100, 10, and 1 pg/μL, and 100, 10, and 1 fg/μL, as well as recombinant pUC57 plasmids containing the target gene fragments at 1 × 10⁵, 1 × 10⁴, 1 × 10³, 1 × 10², 1 × 10¹, 1 × 10⁰ copies/reaction, to determine the analytical sensitivity. Genomic DNA from Ascaris lumbricoides, Ancylostoma duodenale, Enterobius vermicularis, Angiostrongylus cantonensis, Trichinella spiralis, Clonorchis sinensis, Schistosoma japonicum, and Taenia saginata was used to assess assay specificity. A total of 25 stool samples from patients suspected of S. stercoralis infection were tested by the modified Baermann funnel technique, PCR, and the established fluorescent RAA assay. The sensitivity, specificity, concordance rate and their 95% confidence intervals (CI) of these three techniques were estimated, and agreement between methods was evaluated using the Kappa coefficient. Results Exo-4 was identified as the optimal primer set screened from the six primer sets, and the best amplification performance was achieved when the final concentrations of the forward and reverse primers were 0.44 μmol/L and a probe concentration was 0.20 μmol/L. The limit of detection of the fluorescent RAA assay was 100 fg/μL for genomic DNA of S. stercoralis and 1 × 10⁰ copies/reaction for recombinant plasmids. Specific fluorescence signals were detected within 5 min, with no cross-reactivity observed with A. lumbricoides, A. duodenale, E. vermicularis, A. cantonensis, T. spiralis, C. sinensis, S. japonicum, or T. saginata. Among the 25 clinical stool samples from patients suspected of S. stercoralis infections, the modified Baermann funnel technique and fluorescent RAA assay detected 19 positives and 6 negatives, whereas PCR detected 18 positives and 7 negatives. The fluorescent RAA assay showed a sensitivity of 100.00% [95% CI: (82.35%, 100.00%)], specificity of 100.00% [95% CI: (54.07%, 100.00%)], concordance rate of 100.00% [95% CI: (86.28%, 100.00%)], and a Kappa coefficient of 1.00 [95% CI: (1.00, 1.00)] (P < 0.001) relative to the modified Baermann funnel technique, and a sensitivity of 100.00% [95% CI: (81.47%, 100.00%)], specificity of 85.71% [95% CI: (42.13%, 99.64%)], concordance rate of 96.00% [95% CI: (79.65%, 99.90%)], and a Kappa coefficient of 0.90 [95% CI: (0.70, 1.00)] (P < 0.001). Positive amplification products emitted green fluorescence under a portable blue-light device, enabling visual interpretation of results. Conclusions The fluorescent RAA assay established in this study is rapid, highly sensitive, and highly specific. It enables detection of S. stercoralis nucleic acid under isothermal conditions and allows visual interpretation of results, providing a novel tool for rapid clinical diagnosis and field screening of S. stercoralis infections.

Objective : To explore the association between serum γ-aminobutyric acid ( GABA) levels and the risk of developing type 2 diabetes( T2DM) ． Methods : 187 cases of T2DM patients attending the hospital were selected as the T2DM group，and 187 cases of non-T2DM population attending the same period of time were selected as the control group according to age ( ± 3 years) and gender 1 ∶ 1．On-site questionnaires and physical examination were conducted for the study subjects，and serum levels of GABA，Malondialdehyde ( MDA) and activities of superoxide dismutase ( SOD) and Glutathione peroxidase ( GSH-Px) were detected by using ELISA kits．The differences in the levels of GABA and oxidative stress indicators ( SOD，GSH-Px，MDA) between the two groups were compared， and the correlation between GABA and oxidative stress indicators was analyzed by Spearman's method; GABA and oxidative stress indicators were divided into three groups according to their control quartiles，respectively ［low level group ( Q1: ＜P25) ，medium level group ( Q2: P25 －P75) ，high level group ( Q3: ＞P75) ］，and conditional logistic regression was applied to analyze the relationship between GABA，oxidative stress indicators and the risk of develo- ping T2DM; the dose-response relationship between GABA，oxidative stress indicators and the risk of developing T2DM was analyzed by using restricted cubic spline ( RCS) ． Results : T2DM group ( P＜0. 05) ．Spearman's correlation analysis showed that GABA level was positively correlated with SOD and GSH-Px activities and negatively correlated with MDA level ( P＜0. 001) ．Conditional logistic regression analysis showed that medium levels of SOD and GSH-Px as well as medium and high levels of GABA were protective factors for T2DM compared with low levels in each group ( P＜0. 05) ．RCS results showed that a negative dose-response relationship between GABA，GSH-Px and the risk of developing T2DM，and SOD showed a trend of decreasing and then increasing the risk of developing T2DM ( P＜0. 05) ． Conclusion Serum GABA levels have been associated with the risk of developing T2DM．As serum GABA levels increase，the risk of developing T2DM may decrease．

Skeletal muscle dysfunction is a common extrapulmonary comorbidity of chronic obstructive pulmonary disease (COPD) and is associated with decreased quality-of-life and survival in patients. The autophagy lysosome pathway is one of the proteolytic systems that significantly affect skeletal muscle structure and function. Intriguingly, both promoting and inhibiting autophagy have been observed to improve COPD skeletal muscle dysfunction, yet the mechanism is unclear. This paper first reviewed the effects of macroautophagy and mitophagy on the structure and function of skeletal muscle in COPD, and then explored the mechanism of autophagy mediating the dysfunction of skeletal muscle in COPD. The results showed that macroautophagy- and mitophagy-related proteins were significantly increased in COPD skeletal muscle. Promoting macroautophagy in COPD improves myogenesis and replication capacity of muscle satellite cells, while inhibiting macroautophagy in COPD myotubes increases their diameters. Mitophagy helps to maintain mitochondrial homeostasis by removing impaired mitochondria in COPD. Autophagy is a promising target for improving COPD skeletal muscle dysfunction, and further research should be conducted to elucidate the specific mechanisms by which autophagy mediates COPD skeletal muscle dysfunction, with the aim of enhancing our understanding in this field.
Pulmonary Disease, Chronic Obstructive/physiopathology* ; Autophagy/physiology* ; Humans ; Muscle, Skeletal/pathology* ; Mitophagy ; Animals ; Mitochondria/metabolism* ; Lysosomes

Objective To systematically assess the spatiotemporal distribution,risk factors,and future trends of chronic obstructive pulmonary disease(COPD)among individuals under 50 years of age globally from 1990 to 2021 based on Global Burden of Disease(GBD)data in order to provide support for the formulation of prevention and control strategies of the disease.Methods The GBD data from 1990 to 2021 were analyzed for the incidence,mortality,disability-adjusted life years(DALYs),and estimated annual percentage change(EAPC)of COPD in<50-year-old individuals across 204 countries and regions.The data were stratified by age,sex,region,country,and sociodemographic index(SDI).The COPD trends until 2035 were predicted.Results In 2021,the global incidence of early-onset COPD was estimated at 2.5 million cases(95%uncertainty interval:2.09～2.96 million),representing a 50.55%increase compared to 1990.Significant regional heterogeneity was observed,with low SDI regions experiencing a 134.08%increase,whereas high SDI regions exhibited a rise-then-fall trend.Risk factor analysis identified environmental and occupational exposures(air pollution,ambient ozone pollution,household air pollution from solid fuels,etc.)and smoking as the primary etiological factors.Notably,household solid fuel exposure accounted for 50.90%of COPD-related deaths in low SDI regions,compared to only 0.03%in high SDI regions.Projections indicated that by 2035,the global burden of early-onset COPD will increase to 2.59 million cases.Conclusion The global disease burden of COPD among people under 50 years increased significantly from 1990 to 2021,with pronounced disparities across regions and socioeconomic levels.COPD deaths in low-SDI regions are strongly associated with solid fuel exposure and particulate matter pollution,and these regions are expected to remain the main drivers of global COPD incidence growth through 2035.

Objective To explore the effects of commonly used rapid imaging techniques on the image quality of cerebral three-dimensional time-of-flight magnetic resonance angiography(3D-TOF-MRA)and to select the optimal imaging technique for clinical use.Methods Thirty subjects were prospectively recruited,and five sets of 3D-TOF-MRA images were acquired as follows:non-acceleration(reference group),intra-layer parallel imaging(PI)(1D group),inter-layer PI(2D group),compressed sensing(CS)group,and artificial intelligence-assisted compressive sensing(ACS)group.The metrics including clarity,pseudo-stenosis,signal-to-noise ratio(SNR),contrast-to-noise ratio(CNR),and edge sharpness of the intracranial proximal,middle,and distal blood vessels were quantitatively or qualitatively assessed.Results There were no significant differences in the clarity of intracranial proximal and middle blood vessels between the groups(P＞0.05).The clarity of distal blood vessels in the ACS group was superior to that in the other groups,and the CS and 2D groups were superior to the 1D group.Compared to the reference group,no stenosis was observed in any segment of the blood vessels in the ACS group,while the incidence of stenosis in the distal blood vessels of the 1D,2D,and CS groups was 36.67％,13.33％,and 6.67％,respectively.The SNR of the proximal and middle blood vessels in the ACS group did not differ from the reference group,and the SNR of the proximal,middle,and distal segments was higher than that in the 1D and CS groups,and higher than the 2D group for the proximal and distal segments.The CNR of the proximal,middle,and distal segments in the ACS group did not differ from the reference group,and was higher than that of the 1D and CS groups,and higher than the 2D group for the proximal and distal segments.The edge sharpness of the ACS group was significantly higher than that of the other groups.Conclusion ACS technology can accelerate the imaging speed and improve the image quality of cerebral 3D-TOF-MRA.It is recommended for use.

Objective:To explore the latent profile characteristics of fatigue in patients with non-small cell lung cancer (NSCLC) complicated by radiation-induced pulmonary fibrosis (RIPF), and to provide evidence for developing precision nursing strategies.Methods:A convenience sample of 120 patients with RIPF and NSCLC who received treatment at the First Affiliated Hospital of Zhengzhou University between January 2022 and December 2023 was recruited. Baseline demographic and clinical data and the Multidimensional Fatigue Inventory (MFI) were collected. Latent profile analysis (LPA) was performed to classify fatigue levels, and multinomial logistic regression was used to identify influencing factors. A total of 120 questionnaires were distributed, and 116 valid responses were obtained, with a valid response rate of 96.67% (116/120) .Results:LPA identified three latent classes of fatigue among the 116 patients: the physiological-cognitive compound fatigue group ( n=52), the emotional-sleep disturbance group ( n=38), and the mildly adaptive group ( n=26). Multinomial logistic regression revealed that age, Eastern Cooperative Oncology Group performance status (ECOG-PS), Karnofsky Performance Status (KPS), sleep quality, and anxiety were significant factors associated with the physiological-cognitive compound fatigue group ( P<0.05). Sleep quality, anxiety, depression, pain, and KPS were significant factors associated with the emotional-sleep disturbance group ( P<0.05) . Conclusions:Patients with RIPF and NSCLC can be classified into three subtypes of fatigue. Differentiated nursing strategies should be developed accordingly to achieve precise and individualized interventions.

Objective To explore the effects of commonly used rapid imaging techniques on the image quality of cerebral three-dimensional time-of-flight magnetic resonance angiography(3D-TOF-MRA)and to select the optimal imaging technique for clinical use.Methods Thirty subjects were prospectively recruited,and five sets of 3D-TOF-MRA images were acquired as follows:non-acceleration(reference group),intra-layer parallel imaging(PI)(1D group),inter-layer PI(2D group),compressed sensing(CS)group,and artificial intelligence-assisted compressive sensing(ACS)group.The metrics including clarity,pseudo-stenosis,signal-to-noise ratio(SNR),contrast-to-noise ratio(CNR),and edge sharpness of the intracranial proximal,middle,and distal blood vessels were quantitatively or qualitatively assessed.Results There were no significant differences in the clarity of intracranial proximal and middle blood vessels between the groups(P＞0.05).The clarity of distal blood vessels in the ACS group was superior to that in the other groups,and the CS and 2D groups were superior to the 1D group.Compared to the reference group,no stenosis was observed in any segment of the blood vessels in the ACS group,while the incidence of stenosis in the distal blood vessels of the 1D,2D,and CS groups was 36.67％,13.33％,and 6.67％,respectively.The SNR of the proximal and middle blood vessels in the ACS group did not differ from the reference group,and the SNR of the proximal,middle,and distal segments was higher than that in the 1D and CS groups,and higher than the 2D group for the proximal and distal segments.The CNR of the proximal,middle,and distal segments in the ACS group did not differ from the reference group,and was higher than that of the 1D and CS groups,and higher than the 2D group for the proximal and distal segments.The edge sharpness of the ACS group was significantly higher than that of the other groups.Conclusion ACS technology can accelerate the imaging speed and improve the image quality of cerebral 3D-TOF-MRA.It is recommended for use.

Objective:To explore the latent profile characteristics of fatigue in patients with non-small cell lung cancer (NSCLC) complicated by radiation-induced pulmonary fibrosis (RIPF), and to provide evidence for developing precision nursing strategies.Methods:A convenience sample of 120 patients with RIPF and NSCLC who received treatment at the First Affiliated Hospital of Zhengzhou University between January 2022 and December 2023 was recruited. Baseline demographic and clinical data and the Multidimensional Fatigue Inventory (MFI) were collected. Latent profile analysis (LPA) was performed to classify fatigue levels, and multinomial logistic regression was used to identify influencing factors. A total of 120 questionnaires were distributed, and 116 valid responses were obtained, with a valid response rate of 96.67% (116/120) .Results:LPA identified three latent classes of fatigue among the 116 patients: the physiological-cognitive compound fatigue group ( n=52), the emotional-sleep disturbance group ( n=38), and the mildly adaptive group ( n=26). Multinomial logistic regression revealed that age, Eastern Cooperative Oncology Group performance status (ECOG-PS), Karnofsky Performance Status (KPS), sleep quality, and anxiety were significant factors associated with the physiological-cognitive compound fatigue group ( P<0.05). Sleep quality, anxiety, depression, pain, and KPS were significant factors associated with the emotional-sleep disturbance group ( P<0.05) . Conclusions:Patients with RIPF and NSCLC can be classified into three subtypes of fatigue. Differentiated nursing strategies should be developed accordingly to achieve precise and individualized interventions.

Objective:To investigate the therapeutic efficacy of different modes of neoadjuvant therapy for locally advanced rectal cancer (LARC) with proficient in mismatch repair intact (pMMR) or microsatellite stability (MSS).Methods:A retrospective case series study was conducted. A total of 210 LARC patients with pMMR or MSS admitted to the First Affiliated Hospital of Zhengzhou University from January 2020 to December 2022 were selected. Patients were divided into the chemotherapy alone group (98 cases) and the total neoadjuvant therapy (TNT) group (112 cases) based on different neoadjuvant treatment modes. The pathological complete remission (pCR) rate of both groups was compared. In the TNT group, 24 patients received short-course radiotherapy (SCRT) plus chemotherapy (group A), 62 patients received SCRT plus chemotherapy combined with immunotherapy (group B), 23 patients received long-course radiotherapy (LCRT) plus chemotherapy (group C), and 3 patients received LCRT plus chemotherapy combined with immunotherapy (group D). The pCR rate, tumor down-staging and the incidence of adverse reactions were compared between the 4 groups. A subgroup analysis was performed in 87 patients with the distance from the tumor site to anal verge >5 cm and T 2-3N 0-2M 0 TNM staging. And then 87 patients were divided into the chemotherapy alone group (47 cases) and the TNT group (40 cases), and the overall survival (OS) and disease-free survival (DFS) of patients were compared between the 2 groups. Results:Among the 210 LARC patients, 126 cases were male and 84 cases were female, with the age of (51±11) years. There were 3 cases in clinical stage Ⅱ and 207 cases in clinical stage Ⅲ; 68 cases with the distance from the tumor site to the anal verge <5 cm and 142 cases with the distance from the tumor site to the anal verge ≥ 5 cm. The pCR rate of the TNT group was higher than that of the chemotherapy alone group [40.2% (45/112) vs. 7.2% (7/98)], and the difference was statistically significant ( χ2 = 30.62, P < 0.001). In group D, 1 patient achieved pCR, but no statistical comparison was made due to the small sample size. There was no statistically significant difference in pCR rate between group A and group C [25.0% (6/24) vs. 21.7% (5/23), χ2 = 0.07, P = 0.792]. The pCR rate of group B was higher than that of group A and group C [53.2% (33/62) vs. 25.0% (6/24), 53.2% (33/62) vs. 21.7% (5/23)], and the differences were statistically significant ( χ2 = 5.56, P = 0.029; χ2 = 6.73, P = 0.013). However, there were no statistically significant differences in T and N down-staging among group A, group B and group C (all P > 0.05). During neoadjuvant treatment, the incidence of grade 3-4 myelosuppression was 10.7% (12/112) in the TNT group, and no grade 3-4 radiation proctitis was observed. The incidence of granulocytopenia in group B was lower than that in group A, and the incidence of thrombocytopenia in group was lower than that in group C; and the differences were statistically significant (all P < 0.05). In the subgroup analysis, there were no statistically significant differences in OS and DFS between the chemotherapy alone group and the TNT group ( χ2 = 2.17, P = 0.141; χ2 = 0.24, P = 0.624). Conclusions:The neoadjuvant treatment of SCRT combined with immunotherapy has a high pCR rate and a good safety in LARC patients with pMMR or MSS. TNT is not effective in improving the survival of low-risk LARC patients with tumors located in the mid to high segment of the rectum and may have the risk of overtreatment.

Skeletal muscle dysfunction is a common extra-pulmonary complication in patients with chronic ob-structive pulmonary disease(COPD),significantly impacting exercise capacity and quality of life,leading to a poorer prognosis and increased mortality.Oxidative stress closely associates with the development and progression of skeletal muscle dysfunction in COPD.Exercise,a core component of pulmonary rehabilitation,stands as the primary non-pharma-cological treatment for skeletal muscle dysfunction in COPD patients and exerts a positive modulating effect on oxidative stress.This paper reviews the effects of oxidative stress on skeletal muscle dysfunction in COPD and discusses the mecha-nisms by which exercise improves skeletal muscle dysfunction in COPD from an anti-oxidative stress perspective.It has been found that oxidative stress affects the structure and function of muscles in COPD patients by upregulating the protein hydrolysis system,disrupting mitochondrial function,and impairing calcium homeostasis.Mechanisms by which exercise modulates oxidative stress to improve skeletal muscle dysfunction include the activation of antioxidant genes such as silent mating type information regulation 2 homolog 1 and nuclear factor erythroid 2 related factor to enhance the body's antioxi-dant capacity,inhibiting muscle atrophy.Exercise also regulates mitochondrial reactive oxygen species metabolism,im-proving mitochondrial function,and reduces oxidase activity to protect sarcoplasmic reticulum calcium regulation.In con-clusion,the regulation of skeletal muscle oxidative stress by exercise is a crucial target for improving skeletal muscle dys-function in COPD.