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.

ObjectiveTo explore the relationship between triglyceride high-density cholesterol-glucose body index （TyHGB） and gestational diabetes mellitus （GDM）. MethodsThis single-center retrospective cohort study was conducted. A total of 723 singleton pregnant women who received prenatal care and delivered at Lingnan Hospital， the Third Affiliated Hospital of Sun Yat-sen University， from May 2024 to July 2024 were enrolled. Clinical baseline data and laboratory test results in the first trimester （before 13⁺⁶ weeks of gestation） were collected. Based on the tertiles of the TyHGB index were equally divided into 3 groups. Clinical baseline data， and GDM risk were compared among the three groups. ResultsCompared with the control group， the GDM group had older maternal age， higher Pre-pregnancy BMI， FPG， TG， and higher values of 75 g OGTT fasting plasma glucose， 1 h plasma glucose， and 2 h plasma glucose. The early-pregnancy TyHGB index， andincidence of macrosomiawere all higher in the GDM group than in the control group （ all P<0.05）， whereas gestational weight gain （GWG） was lower than that of the control group （P<0.05）. The individuals were divided into three groups according to the tertiles of the TyHGB index. Significantly differences were identified among TyHGB index tertiles in maternal age， parity， Pre-BMI， FPG， TC， TG， HDL-C， LDLL-C， Ur， 75 g OGTT fasting plasma glucose， 1 h plasma glucose， 2 h plasma glucose， GWG， neonatal birth weight， GDM， GH， macrosomia incidence， and cesarean section rate（all P<0.05）. Further trend test analysis found that matenal age， parity， Pre-BMI， FPG， TC， TG， LDL-C， 75 g OGTT_FPG， 75 g OGTT_1 h PG and 75 g OGTT_2 h PG level， neonatal birth weight， GDM， GH， incidence of macrosomia and cesarean section rate all showed an upward trend. The multivariate Logistic regression models revealed that there was an independent positive correlation between TyHGB index and the risk of GDM. Curve fitting by restricted cubic splines （RCS） were used to assess linearity between TyHGB and GDM， and there was a linear dose-response relationship between TyHGB and GDM （P _{nonlinear test} = 0.597）. The threshold value for early pregnancy TyHGB index to predict GDM was 6.449 ， with an area under the curve （AUC） of 0.644 （95% CI： 0.597-0.691）， sensitivity of 0.480 ， and specificity of 0.779. After adjusting for maternal age， Pre-BMI， family history of diabetes， and history of GDM， the AUC for the early pregnancy TyHGB index to predict GDM was 0.672 （95% CI： 0.626-0.718）. ConclusionThe early-pregnancy TyHGB index is independently associated with GDM risk， and exhibits a linear dose-response relationship with GDM. It may serve as an early screening and monitoring tool for pregnant women at high risk of GDM. However， its predictive value for GDM requires further validation.

Objective:To observe the effects of Yiyuan moxibustion on bladder function and antioxidant level of spinal cord tissues in rats with urinary retention after spinal cord injury (SCI); To explore the mechanism of inhibition of ferroptosis in spinal cord neurons after SCI by Yiyuan moxibustion.Methods:Wistar female rats were divided into sham-operation group, model group, and Yiyuan moxibustion group according to random number table method, with 12 rats in each group. The modified Allen′s vertical percussion method was used to construct the model of urinary retention after SCI in T10 segment. The rats in the Yiyuan moxibustion group were moxibued at the Zhongji acupoint, Guanyuan acupoint, and Shenque acupoint for 20 min per day, and the intervention was continued for 2 weeks. Urodynamic test was used to observe the degree of urinary retention in rats; HE staining was used to observe the morphological changes of the injured spinal cord tissues; transmission electron microscopy was used to observe the mitochondrial ultrastructure of the spinal cord tissues; ferric ion kit was used to detect the ferric ion content of the spinal cord tissues; ELISA was used to detect the GSH and MDA contents of the spinal cord tissues of the rats; Western blot was used to measure the relative expressions of glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) proteins in rat spinal cord tissue.Results:Compared with the model group, the basal and leakage point pressures of the bladder, and bladder compliance were significantly reduced in the Yiyuan moxibustion group ( P<0.05); the spinal cord tissue structure was restored and mitochondrial morphology improved; the levels of iron ions and MDA in spinal cord tissue decreased ( P<0.05), while the level of GSH increased ( P<0.05), and the relative expressions of GPX4 and SLC7A11 proteins increased ( P<0.05). Conclusion:Yiyuan moxibustion can improve bladder function in rats with urinary retention after SCI, and the mechanism may involve the initiation of antioxidant defense and reduction of lipid peroxidation in spinal cord neuronal cells, thus preventing the occurrence of ferroptosis and achieving the protection of neuronal cells.

Objective To explore the value of combined detection of serum mitogen-activated protein kinase 1(MAPK1)and lysyl oxidase-like protein 2(LOXL2)in early diagnosis of cervical cancer.Methods A total of 218 patients with cervical lesions were selected as study group(103 ca-ses in cervical cancer group,115 cases in benign tumor group).Additionally,100 patients with cer-vical intraepithelial neoplasia grade Ⅱ were selected as precancerous lesion group,and 79 healthy in-dividuals undergoing physical examinations during the same period were selected as control group.Se-rum levels of MAPK1 and LOXL2 were measured in each group.Pearson correlation analysis was used to evaluate the correlations of serum MAPK1 and LOXL2 levels in patients with cervical cancer.Logistic regression analysis was performed to screen influencing factors for the occurrence of cervical cancer.Receiver operating characteristic(ROC)curves were plotted to assess the diagnostic efficacy of serum MAPK1 and LOXL2 for cervical cancer.Results Serum MAPK1 and LOXL2 levels in the study group were higher than those in the precancerous lesion group and the control group,and those in the precancerous lesion group were higher than those in the control group,with statistically signif-icant differences(P＜0.05).The proportion of patients with high-risk human papillomavirus(HPV)infection and serum MAPK1 and LOXL2 levels in the cervical cancer group were higher than those in the benign tumor group,with statistically significant differences(P＜0.05).Serum MAPK1 and LOXL2 levels in patients with stage Ⅲ to Ⅳ cervical cancer were higher than those in patients with stage Ⅰ to Ⅱ cervical cancer,with statistically significant differences(P＜0.05).Pearson correlation analysis showed a positive correlation between serum MAPK1 and LOXL2 levels in patients with cervical cancer(r=0.468,P＜0.001).Logistic regression analysis showed that high-risk HPV infection,MAPK1 and LOXL2 were all influencing factors for the occurrence of cervi-cal cancer(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)for com-bined diagnosis of serum MAPK1 and LOXL2 was 0.911,which was significantly greater than the AUCs for individual diagnoses(0.848 and 0.843,respectively).Conclusion Serum MAPK1 and LOXL2 levels in patients with cervical cancer are significantly upregulated,and the two indicators were positively correlated.High-risk HPV infection,serum MAPK1 and LOXL2 levels were influen-cing factors for the occurrence of cervical cancer.Combined detection of MAPK1 and LOXL2 levels is expected to assist in the diagnosis of cervical cancer.

OBJECTIVE: To explore the pathogenic mechanism of a child with Waardenburg syndrome type 4C due to a c.661_664dup (p.P222Lfs*60) variant of SOX10 gene through in vitro experiments. METHODS: A child diagnosed at the Handan First Hospital was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples were collected from the child and his parents. Following extraction of genomic DNA, trio-whole exome sequencing was carried out. Pathogenicity of candidate variant was determined by bioinformatic analysis and reference to the guidelines from the American College of Medical Genetics and Genomics (ACMG). Candidate variant was verified by Sanger sequencing. Expression plasmids of wild-type SOX10 and the c.661_664dup (p.P222Lfs*60) variant were constructed and transiently transfected into 293T cells to determine the expression at the RNA and protein levels. The 293T cells transiently transfected with the wild-type/mutant SOX10 were treated with 10 ug/mL cycloheximide (CHX) for 0, 4, 8, 24 h, respectively, and the degradation rate of target protein was detected by Western blotting assay. This study has been approved by the Ethics Committee of Handan First Hospital (Ethics No. HDYY-LW-25053). RESULTS: The child was found to harbor a heterozygous c.661_664dup (p.P222Lfs*60) variant of the SOX10 gene, which was unreported previously. The variant did not significantly alter the expression of SOX10 at the mRNA level but the protein level. After the CHX treatment, the degradation of mutant SOX10 protein had slowed down. CONCLUSION The mutant SOX10 may affect the expression of downstream genes by affecting the degradation rate of its protein product.
Humans ; HEK293 Cells ; Mutation ; SOXE Transcription Factors/metabolism* ; Waardenburg Syndrome/genetics* ; Child

Objective:To explore the dynamic change trajectory and influencing factors of postoperative fatigue (POF) in patients with early digestive tract cancer undergoing endoscopic submucosal dissection (ESD), so as to provide theoretical basis for individualized management of such patients.Methods:Using cross-sectional survey and convenient sampling method, the patients who underwent ESD for early cancer of digestive tract in the endoscopic center of the Second Affiliated Hospital of Zhejiang University School of Medicine from January to June 2024 were selected as the research objects. The questionnaires were conducted at 30 min, 24 h, 3 days, 5 days, 7 days after ESD with Christensen's postoperative fatigue score. Spearman test was used to analyze the correlation between pain, insomnia and POF. The latent variable growth model was used to identify the potential categories of POF trajectory, and the influencing factors were analyzed by Logistic regression.Results:A total of 232 patients were finally induded, with ages of 19-94(59.53 ± 13.29) years, including 120 males and 112 femalss. The POF level of patients with early cancer of digestive tract showed a downward trend one week after operation. Three postoperative fatigue trajectories were fitted, 9.05% in moderate or severe fatigue low-speed smooth descent group (C1 group); 32.76% in moderate fatigue first decreased quickly and then slowly group (C2 group) and 58.19% in mild fatigue continuous decline group (C3 group). The result of Logistic regression analysis showed that, compared with C1 group, people without religious beliefs were morelikely to enter C2 group [ β = 1.572, OR = 4.818(1.033 - 22.465), P<0.05]; compared with C3 group, patients with high pain level and severe insomnia degree were likely to enter C1 [ β = 2.621, 0.663, OR were 13.754(2.692 - 70.283) and 1.942(1.429 - 2.638), both P<0.05] and C2 [ β = 2.010, 0.491, OR were 7.464(1.890 - 29.482) and 1.634(1.348 - 1.982), both P<0.05] group. Conclusions:There were three potential types of POF in patients with early digestive tract cancer ESD. Medical staff should pay attention to patients with severe insomnia, intense pain and religious beliefs, and give staged fatigue assessment and individualized intervention, so as to reduce the level of POF and promote rapid recovery.

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 explore the pathogenic mechanism of a child with Waardenburg syndrome type 4C due to a c. 661_664dup (p.P222Lfs*60) variant of SOX10 gene through in vitro experiments. Methods:A child diagnosed at the Handan First Hospital was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples were collected from the child and his parents. Following extraction of genomic DNA, trio-whole exome sequencing was carried out. Pathogenicity of candidate variant was determined by bioinformatic analysis and reference to the guidelines from the American College of Medical Genetics and Genomics (ACMG). Candidate variant was verified by Sanger sequencing. Expression plasmids of wild-type SOX10 and the c. 661_664dup (p.P222Lfs*60) variant were constructed and transiently transfected into 293T cells to determine the expression at the RNA and protein levels. The 293T cells transiently transfected with the wild-type/mutant SOX10 were treated with 10 μg/mL cycloheximide (CHX) for 0, 4, 8, 24 h, respectively, and the degradation rate of target protein was detected by Western blotting assay. This study has been approved by the Ethics Committe of Handan First Hospital(Ethics No.HDYY-LW-25053). Results:The child was found to harbor a heterozygous c. 661_664dup (p.P222Lfs*60) variant of the SOX10 gene, which was unreported previously. The variant did not significantly alter the expression of SOX10 at the mRNA level but the protein level. After the CHX treatment, the degradation of mutant SOX10 protein had slowed down. Conclusion:The mutant SOX10 may affect the expression of downstream genes by affecting the degradation rate of its protein product.

Based on the analysis of relevant policy documents,industry data,and case studies,this paper systematically reviews the current situation,policy environment,and usage of intra-hospital formulations in medical institutions in Guangzhou,and compares them with policies in other countries.It is found that although Guangzhou has achieved certain results in terms of policy support,number of varieties,and usage range of intra-hospital formulations,there are also problems such as limited pro-duction capacity,inadequate quality control,insufficient research and development investment,increasing cost pressure,and in-adequate protection of intellectual property rights.Therefore,this paper suggests the establishment of a regional Chinese medicine formulation industrial park,the development of new dosage forms and technologies,the expansion of medical insurance coverage,the strengthening of research platform construction,and the promotion of multidimensional achievement transformation to promote the high quality development of the intra-hospital formulation industry in Guangzhou.

To provide in-service medical technicians and nurses with convenient access to continuing education resources in transfusion medicine, reduce transfusion-related adverse events, and ensure the safety, rationalization, and effectiveness of clinical transfusion, we designed and developed an online transfusion continuing education platform. The platform was based on the new managed code programming model.NET Core and the powerful functions of hypertext preprocessor PHP 7.4, addressing current issues in transfusion online continuing education. Through in-depth analysis of student attributes, learning behaviors, and teaching behaviors, a comprehensive online continuous teaching quality evaluation index system was established. This system not only facilitates the quantitative assessment of teaching quality but also successfully integrates the two core functions of teaching and management, thereby achieving unified online teaching.