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 effect of vaginal sacrospinous ligament suspension(VSSLS)with uterus preservation in treatment of patients with moderate to severe pelvic organ pro-lapse(POP).Methods A total of 92 patients with moderate to severe POP were randomly divided into control group and observation group,with 46 cases in each group.The control group received va-ginal anterior and posterior wall repair(VAPWR),while the observation group underwent VSSLS.The intraoperative blood loss,operation time and hospital stay were compared between the two groups;the Pelvic Floor Distress Inventory-20(PFDI-20)was used to evaluate quality of life;the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire-12(PISQ-12)was used to assess conditions of sexual satisfaction degree,dyspareunia,sexual frequency,and perceived control during sexual activi-ty;surgical complications in both groups were recorded.Results The observation group had less intr-aoperative blood loss,shorter operation time,and shorter hospital stay compared with the control group(P＜0.01).After treatment,the observation group had lower scores in all dimensions of the PFDI-20 compared with the control group,and the decreasing range was significantly greater than that in the control group(P＜0.01).After treatment,the observation group had significant higher scores for sex-ual satisfaction degree,sexual frequency and perceived control during sexual activity compared with the control group,and the change in these indicators before and after treatment was also significantly greater in the observation group,while the score for dyspareunia was significantly lower in the observation group,with a significant decrease compared with the control group(P＜0.01).The incidence of complications was significantly lower in the observation group than that in the control group(P＜0.01).Conclusion As a surgical approach for the treatment of moderate to severe POP,VSSLS can significantly improve patient's quality of life.

Objective To compare the application value of indocyanine green(ICG),nano-car-bon,methylene blue(MB)tracing methods,and ICG combined with nano-carbon tracing method in laparoscopic surgery for stage Ⅰ to Ⅱ endometrial cancer according to International Federation of Gynecology and Obstetrics(FIGO)staging system.Methods A retrospective selection of 92 pa-tients who underwent laparoscopic surgery for stage Ⅰ to Ⅱ endometrial cancer was conducted.Based on intraoperative sentinel lymph node(SLN)tracing method,the patients were divided into ICG group(n=24,using ICG tracing method),nano-carbon group(n=22,using nano-carbon tracing meth-od),MB group(n=23,using MB tracing method),and combined group(n=23,using ICG combined with nano-carbon tracing method).Surgical-related indicators,pathological results,and the incidence of complications were compared among the four groups.The detection efficacy of different tracing methods was also compared,using pathological results of surgical specimens as the gold standard.Results There were no statistically significant differences in the SLN resection time,to-tal operative time,and intraoperative blood loss among the four groups(P＞0.05).The number of lymph nodes resected intraoperatively in the combined group was significantly higher than that in the other three groups(P＜0.05).There were no statistically significant differences in the pathological results of surgical specimens(tumor invasion depth＞50％,tumor diameter＞2 cm,tumor invasion of the cervix or lymphovascular space)among four groups(P＞0.05).The complication rates in the ICG group,nano-carbon group,MB group,and combined group were 4.17％,9.09％,8.70％,and 13.04％,respectively,with no statistically significant difference(P＞0.05).Using the pathological results of surgical specimens as the gold standard,the detection accuracy,sensitivi-ty,and specificity of ICG combined with nano-carbon tracing method for SLNs in patients with stage Ⅰto Ⅱ endometrial cancer were 86.96％,90.00％,and 66.67％,respectively.The positive predictive value and negative predictive value were 94.74％and 50.00％,respectively.The detection efficacy of this combined method was higher than that of the other three methods.Conclusion Compared with the ICG,nano-carbon,and MB tracing method,ICG combined with nano-carbon tracing method has a higher application value in laparoscopic surgery for stage Ⅰ toⅡ endometrial cancer.It can accurately i-dentify the status of lymph node metastasis,with a moderate price and simple operation.

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.

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.

Objective　To investigate the contamination status of Bacillus cereus in food, the profile of virulence factors carried, and drug sensitivity characteristics in Liaoning Province. Methods　A total of 620 samples were isolated from 15 monitoring sites, covering four major sample types of food products in Liaoning Province between 2021 and 2022. Bacillus cereus was isolated and identified by GB4789.14—2014, and its virulence was assessed using PCR detection of the groEL and gyrB genes specific to the bacterial group, as well as 11 other virulence genes. Broth microdilution method was used to detect the antibiotic sensitivity of the strain. Results　All 79 Bacillus cereus strains were found to carry virulence genes. The detection rates of non-hemolytic enterotoxin genes nheC, nheA, and nheB were 100% (79/79), 93.3% (74/79), and 83.5% (66/79), respectively. The detection rate of enterotoxin entFM gene was 98.7% (78/79). The 79 strains of Bacillus cereus carried 19 types of virulence gene patterns, with IX and XVII gene patterns being the most prevalent, each accounted for 16.5% (13/79). The resistance rates of 79 strains of Bacillus cereus to 10 antibacterial drugs were 100% for both ampicillin and penicillin. The resistance rates to cotrimoxazole, imipenem, erythromycin, clindamycin, ciprofloxacin, and tetracycline 65.8% (52/79), 10.1% (8/79), 73.4% (58/79), 60.8% (48/79), 75.9% (60/79), and 70.9% (56/79) respectively. The resistance rates to chloramphenicol and gentamicin were both 0%. Conclusion　Food from catering services in Liaoning Province shows contamination by Bacillus cereus, with isolated strains carrying at least four kinds of virulence genes. About 100% of the isolates were resistant to ampicillin and penicillin, whereas high sensitivity was observed to chloramphenicol and gentamicin. Active monitoring, timely targeted prevention and control interventions should be strengthened to provide a theoretical basis for future traceability studies.

Objective To screen the differentially expressed proteins in glomeruli during the treatment of lupus nephritis(LN)with hydroxychloroquine sulfate(HCQ).Methods Sixty female NZB/WF1 mice were used.At the age of 28 weeks,40 mice with proteinuria of 3+to 4+were divided into HCQ and control groups.After feeding for 36 weeks,the glomerular proteins were extracted by magnetic beads and analyzed by two-dimensional fluorescence difference gel electrophoresis(2D-DIGE)and matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry(MALDI-TOF-MS).The expression of the candidate proteins in the glomeruli of the LN mice was exa-mined by immunohistochemistry.Results A total of 31 differentially expressed proteins were identified between the two groups,including 24 upregulated and seven downregulated proteins.Conclusion The expression of proteins like RI and ENOA in the glomeruli of LN mice was significantly different during HCQ treatment.These proteins may be involved in the pathogenesis of LN and are therefore potential targets of HCQ in the treatment of LN.

Objective:Analyze the impact of smoking on the mortality and life expectancy of residents in Tianjin in 2019.Methods:Use mortality case-control study method to collect all cause of death cases of residents in Tianjin in 2019 for analysis. After adjusting for the 5-years-old age group, education level, and marital status, the smoking attributed deaths from different diseases of different genders, smoking attributed deaths in different age groups, and their impact on life expectancy were analyzed.Results:The total number of deaths in 2019 was 75 254, with 42 201 males (56.1%). Among male deaths, 3 215 (9.9%) were attributed to smoking, of which 2 157 (50.2%) lung cancer deaths were attributed to smoking; The risk of lung cancer death among smokers was 3.075 times higher than that of non-smokers (95% CI: 2.812-3.364); Among the 33 053 female deaths (43.9%), 1 396 (5.8%) were caused by smoking, with 744 (29.1%) lung cancer deaths attributed to smoking. The age group with the highest number of deaths attributed to smoking for women was the 75-<80 years old age group, followed by the 70-<75 and 80-<85 years old age groups. The age group with the highest proportion of deaths attributed to smoking for men was the 55-<60 years old age group. In addition, smoking accounts for more than 60% of deaths in the 60-<65, 45-<50, 55-<60, and 65-<70 years old age groups. In 2019, the loss of life expectancy attributed to smoking deaths among all residents in Tianjin was 1.13 years, with a loss of 1.15 years for males and 0.57 years for females. The expected life expectancy excluding deaths caused by smoking was 82.92 years, 80.77 years for males and 84.61 years for females. Conclusions:Smoking remains one of the important risk factors for death among residents. Promoting effective measures to reduce smoking rates is an effective way to increase life expectancy.

Objective: To investigate GATA3 expression and the regulatory mechanism of m6A modification in the re- sponse of alveolar epithelial cells to radiation, and to provide a new therapeutic target for radiation-induced lung injury based on its pathogenesis. Methods: Human lung epithelial cell line (A549) and mouse lung epithelial cell line (MLE-12) were exposed to X-ray irradiation with a single dose of 10 Gy (dose rate 1 Gy/min) and 6 Gy (dose rate 0.75 Gy/min), respect- ively. The expression of VIRMA gene (RNA methylase) was inhibited by lipofection of A549 cells and MLE-12 cells with shRNA-VIRMA plasmid and siRNA-VIRMA interfering fragment, respectively. Quantification of m6A RNA methylation was performed by colorimetry. Changes in the expression of mRNAs of VIRMA, GATA3, and epithelial-mesenchymal transition (EMT) markers in irradiated A549 and MLE-12 cells were determined by qRT-PCR. Changes in the expression of VIRMA,  GATA3,  and  EMT  marker  proteins  in  irradiated  A549  and  MLE-12  cells  were  determined  by  Western  blot. Results: Radiation up-regulated the expression of methylase VIRMA in A549 and MLE-12 cells, which in turn enhanced the m6A of total RNA and the expression of GATA3 gene and protein, resulting in EMT. Furthermore, in A549 and MLE-12 cells, interference of the VIRMA gene significantly reduced the expression of GATA3 gene and protein and the expression of EMT-related molecules. Conclusion　 Radiation induces m6A modification in alveolar epithelial cells, which up-regu- lates the expression of GATA3 gene and induces EMT, thus playing an important role in the process of radiation-induced lung injury.

Infection is one of the most common causes of death in patients with multiple myeloma, which can lead to early death. The incidence of infection in patients with (multiple myeloma, MM) complicated with diabetes is further increased. Proteasome inhibitors, immunomodulatory drugs and other new drug therapy improve the prognosis of MM, but glucocorticoids used in combination chemotherapy are easy to induce glucocorticoid diabetes, lead to elevated blood glucose, increase the risk of MM infection, and affect the prognosis of patients. This article reviews the relationship between infection and disease progression in patients with MM complicated with diabetes mellitus, infection-related factors, and prevention and treatment measures of different types of pathogen infection.