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.

In drug clinical trials,the protection of subjects'privacy should take precedence over scientific exploration and societal benefits.This paper compared the differences in legal systems for the protection of subjects'privacy in China and abroad by employing methods such as literature review,case analysis,and comparison of domestic and foreign situations.It also elucidated the current ethical challenges faced by the protection of subjects'privacy,which primarily included imperfect laws and regulations,insufficient protection awareness among relevant personnel,risks of privacy breaches in critical links,and increased difficulty in data confidentiality under emerging technologies.From the perspective of responsible innovation,specific paths were proposed to protect subjects'privacy by using the four-dimensional structure of"prediction-reflection-negotiation-feedback,"maintaining a balance between ethical principles and technological innovation,ensuring the ethical and compliant nature of drug clinical trials,and thus better contributing to the cause of human health.

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.

In drug clinical trials,the protection of subjects'privacy should take precedence over scientific exploration and societal benefits.This paper compared the differences in legal systems for the protection of subjects'privacy in China and abroad by employing methods such as literature review,case analysis,and comparison of domestic and foreign situations.It also elucidated the current ethical challenges faced by the protection of subjects'privacy,which primarily included imperfect laws and regulations,insufficient protection awareness among relevant personnel,risks of privacy breaches in critical links,and increased difficulty in data confidentiality under emerging technologies.From the perspective of responsible innovation,specific paths were proposed to protect subjects'privacy by using the four-dimensional structure of"prediction-reflection-negotiation-feedback,"maintaining a balance between ethical principles and technological innovation,ensuring the ethical and compliant nature of drug clinical trials,and thus better contributing to the cause of human health.

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.

Thyroid cancer is the most common malignant tumor of the head and neck, among which papillary thyroid carcinoma is the most common. Papillary thyroid carcinoma with a diameter of ≤ 1.0 cm is called thyroid micropapillary carcinoma. In recent years, thermal ablation technology for the treatment of thyroid micropapillary carcinoma has developed rapidly at home and abroad. At present, many guidelines, consensus and clinical studies related to thermal ablation treatment of thyroid micropapillary carcinoma have been published at home and abroad. Based on the existing literature, guidelines and clinical studies, this article summarizes, discusses and analyzes the advantages, indications, efficacy, safety, and existing problems of thermal ablation therapy for thyroid cancer.

Chronic obstructive pulmonary disease(COPD)has become a common chronic disease in the adult in recent years,and more attention has been gradually paid to its prevention and treatment.This paper reviewed the clinical studies about the effect of traditional Chinese medicine(TCM)exercise therapy on COPD,and indicated that TCM exercise therapy can improve the rehabilitation of COPD patients.TCM exercise therapies,such as Tai Chi,Wuqinxi,Baduanjin,and Liuzijue,have been shown to significantly improve lung function,inflammation levels,and exercise capacity in COPD patients in recent studies.Tai Chi significantly improves patient's respiratory problems although lung function indices are not changed.Liuzijue enhances the strength and endurance of respiratory muscles and limbs.Baduanjin helps to improve patient's cognitive and emotional states.Overall,TCM exercise therapy provides an effective rehabilitation option for COPD patients.However,more clinical controlled trials are needed to further confirm their effectiveness and to develop appropriate rehabilitation programs for COPD patients.

P21-activated kinase 5 (PAK5) belongs to the PAK-II subfamily, which is an important regulator of cell survival, adhesion, and motility. However, the functions of PAK5 in endometriosis remain unclear. Here, PAK5 is strikingly upregulated in endometriosis. Furthermore, the knockdown of PAK5 or its inhibitor GNE 2861 blocks the development of endometriosis, which is equally demonstrated in PAK5-knockout mice. In addition, PAK5 promotes glycolysis by enhancing the protein stability of pyruvate kinase 2 (PKM2) in endometriotic cells, which is a key enzyme for glucose metabolism. Moreover, the phosphorylation of PKM2 at Ser519 by PAK5 mediates endometriosis cell proliferation and metastasis. Collectively, PAK5 plays an indispensable role in endometriosis. Our findings demonstrate that PAK5 is an important target for the treatment of endometriosis.
Endometriosis/genetics* ; Female ; Animals ; p21-Activated Kinases/genetics* ; Mice ; Phosphorylation ; Glycolysis ; Humans ; Thyroid Hormone-Binding Proteins ; Membrane Proteins/genetics* ; Carrier Proteins/genetics* ; Cell Proliferation ; Mice, Knockout ; Thyroid Hormones/metabolism* ; Pyruvate Kinase/genetics*

Objective To investigate the effects of insulin therapy on the mechanical behavior of solids,characteristics of fluid flow,and bone marrow stromal cells(BMSCs)differentiation in the distal femoral cancellous bone of type 2 diabetic rats under normal activity and vigorous exercise conditions.Methods The finite element models of cancellous bones and fluids in the distal femurs of rats in the control,diabetes,treatment,and placebo groups in 4-week and 8-week insulin treatment experiments under normal activity and vigorous exercise conditions were established based on micro-CT scanning images.The mechanical and cell differentiation parameters of the models in each group were analyzed using the fluid-solid interaction numerical simulation method.Correlations between mechanical,cell differentiation,and microstructural morphology parameters were also analyzed.Results Insulin therapy under normal activity and vigorous exercise conditions improved the solid and fluid mechanical parameters and BMSC differentiation parameters in type 2 diabetic rats.In the 4-week experiment,insulin treatment under normal activity and vigorous exercise conditions increased the differentiation areas of bone in type 2 diabetic rats from 64.024%to 69.372%and from 73.225%to 75.336%,respectively;in the 8-week experiment,insulin treatment under normal activity and vigorous exercise conditions increased the differentiation areas of bone in type 2 diabetic rats from 67.239%to 72.910%and from 76.147%to 78.291%,respectively.Morphological parameters BV/TV,Tb.N,Tb.Th,Tb.Sp,and structure model index were significantly correlated with the differentiation areas of the bone and cartilage(P<0.05).Conclusions Under vigorous exercise conditions,BMSCs on the surface of cancellous bone in the 8-week insulin treatment group were more likely to differentiate into bone tissue.This study is of great significance for further understanding the effects of insulin on the bone under normal activity and vigorous exercise conditions,and provides theoretical guidance for the selection of the insulin therapy cycle and exercise mode in the clinical treatment of type 2 diabetes.

Bovine leukemia virus(BLV)mainly causes bovine B-cell malignant lymphoma,which causes immunosuppression and leads animals susceptible to other diseases.BLV has been detected all over the world except in Western Europe.Antibodies to BLV have been detected in humans,and reverse transcribed DNA of BLV were detected in breast tissue sections and lung tumors,which in-dicate that BLV is a potential zoonotic disease.At present,there are no effective treatment methods or commercial vaccines for BLV.An in-depth understanding of the development of BLV detection technology will help researchers choose suitable detection methods,avoid invalid detection results,and meet the requirements of disease prevention and control.In view of this,this paper reviews 19 detection technologies of BLV including cytology detection,nucleic acid detection,and serology de-tection.The advantages,disadvantages,and scope of application were also analyzed,which provide theoretical guidance for the prevention and control of bovine leukemia.