Objective To investigate the effects of indirect moxibustion on the expressions of DNA methyltransferases(DNMT)and methylation of the brain-derived neurotrophic factor(BDNF)promoter region in uterine tissues of rats with primary dysmenorrhea(PD);To explore the mechanism of epigenetic regulation of indirect moxibustion on PD model rats.Methods A total of 32 female SD rats were randomly divided into blank group,model group,indirect moxibustion group and Western medicine group,with 8 rats in each group.The PD model with cold dampness stagnation syndrome was established using ice-water baths combined with estradiol benzoate and oxytocin.Starting from the first day of modeling,the indirect moxibustion group received salt-partitioned moxibustion at"Shenque"and ginger-partitioned moxibustion at"Guanyuan"for 20 min,while the Western medicine group was gavaged ibuprofen solution.Both interventions were given once a day for 10 days.On day 11,writhing responses were observed and scored after oxytocin injection,Western blot and RT-qPCR were used to detect protein and mRNA expression of BDNF,DNMT3A and DNMT3B in uterine tissue,immunohistochemical staining was used to detect the positive expressions of DNMT3A and DNMT3B in uterine tissue.The DNA methylation of BDNF promoter region in uterine tissue was detected by sulfite sequencing.Results Compared with the blank group,the writhing latency was shortened and the writhing score increased in the model group(P<0.01);the protein and mRNA expressions of BDNF,DNMT3A and DNMT3B in uterine tissue increased(P<0.01),the positive expressions of DNMT3A and DNMT3B increased(P<0.01),and the DNA methylation rate in BDNF promoter region decreased(P<0.01).Compared with the model group,the writhing latency was lengthened and the writhing score decreased in the indirect moxibustion group and Western medicine group(P<0.05,P<0.01);the protein and mRNA expressions of BDNF,DNMT3A and DNMT3B in uterine tissue decreased(P<0.05,P<0.01),the positive expressions of DNMT3A and DNMT3B decreased(P<0.01),and the DNA methylation rate in BDNF promoter region increased(P<0.01).Conclusion Indirect moxibustion at"Shenque"and"Guanyuan"may inhibit the transcription of BDNF by increasing the DNA methylation level of BDNF promoter region,and reduce the expression of BDNF,so as to relieve the pain of PD rats.

Objective:To evaluate the efficacy of cyclophosphamide in patients with T-cell large granular lymphocytic leukemia (T-LGLL) and the maintenance of treatment-free remission (TFR) following drug discontinuation.Methods:Clinical data were collected from 37 patients with T-LGLL who received oral cyclophosphamide at the Regenerative Medicine Clinic of the Institute of Hematology and Blood Diseases Hospital between June 2019 and March 2024. Patient clinical characteristics, treatment efficacy, and long-term TFR were analyzed.Results:The median age of the 37 patients was 60 years (range: 37-86), and 22 (59.5%) were male. Anemia was observed in 30 patients (81.1%), and 28 (75.7%) met the diagnostic criteria for secondary pure red cell aplasia. Neutropenia occurred in 15 patients (40.5%), lymphocytosis in 11 (29.7%), and thrombocytopenia in three (8.1%). Sixteen patients (43.2%) had not received prior immunosuppressive therapy (treatment-naive group), while 21 patients (56.8%) were refractory to or had relapsed after immunosuppressive treatment (refractory/relapsed group). All patients met the treatment criteria and received oral cyclophosphamide at doses of 50-100 mg/day. Among the 36 evaluable patients, hematologic remission was achieved in 25 (69.4%), with a median time of 2.0 months (range: 0.7-7.0). There was no statistically significant difference in remission rates between the treatment-naive and refractory/relapsed groups (68.5% vs. 66.7%, P=0.589). Among the 25 patients who achieved hematologic remission, 24 discontinued cyclophosphamide. With a median follow-up of 39.0 months (range: 8.0-56.0), the median TFR duration was not reached. The estimated TFR rates were (90.87± 6.16) % at 12 months and (75.72±11.04) % at 36 months. No significant difference in TFR was observed between the treatment-naive and refractory/relapsed groups ( P=0.451) . Conclusion:Oral cyclophosphamide is effective in the treatment of T-LGLL, and patients may maintain long-term TFR following drug discontinuation.

Objective:To evaluate the clinical value of high-risk human papillomavirus (HPV) viral load for the cervical cancer screening and triage of high-risk HPV positive populations without additional tests.Methods:(1) This study conducted a retrospective analysis of 29 720 women aged 35-64 years who received cervical cancer screening in Quanzhou, China, in 2021. Fourteen high-risk HPV types (including HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) were detected for cervical cancer primary screening using hybrid capture-chemiluminescence method. High-risk HPV positive samples were further subjected to HPV 16/18 genotyping using hybrid capture-chemiluminescence method. Among them, HPV 16/18 positive women were directly referred to colposcopy, while the other 12 high-risk HPV positive samples were further subjected to liquid based cytology test. Those with abnormal or suspicious cytology were referred to colposcopy. Biopsies were taken for histopathological examination of suspicious or abnormal individuals under colposcopy. (2) Ten cases of colposcopy loss or refusal to undergo examination were excluded, and the data from the 29 710 cases were analyzed. The HPV viral loads of the other 12 high-risk HPV positive populations were focused and evaluated their HPV viral loads for further cervical intraepithelial neoplasia (CIN) Ⅱ and above lesions (CINⅡ +) triage in cervical cancer screening. Results:(1) Among 29 720 women, 2 487 women (8.37%, 2 487/29 720) were positive for high-risk HPV, including 807 women (2.72%, 807/29 720) were positive for HPV 16/18 and 1 680 patients (5.65%, 1 680/29 720) were positive for the other 12 high-risk HPV types. Among 1 680 women who tested positive for the other 12 high-risk HPV types, 573 patients were atypical squamous cell carcinoma of unclear significance or above, 346 patients were CIN Ⅰ, 122 patients were CIN Ⅱ-Ⅲ, 9 patients were squamous cell carcinoma patients, and 4 patients were adenocarcinoma in situ. The immediate risk of CIN Ⅱ + in HPV 16/18 positive women (11.13%) was approximately four times higher than that of other 12 high-risk HPV positive women (2.74%). (2) Through the viral load analysis of the other 12 high-risk HPV types, we found that the viral load of the other 12 high-risk HPV provide a good value for the pathological results, with a clinical cutoff (CO) value of 11.21 relative light unit/CO (RLU/CO) for the CINⅡ + detection. Except for HPV 16/18 positive patients, when the viral load values of the other 12 high-risk HPV types were greater than 10 RLU/CO, these patients had a higher risk of CINⅡ +, with a positive predictive value of 31.29%. CINⅡ + was not found in any of the other 12 high-risk HPV positive with viral load values less than or equal to 10 RLU/CO. Conclusions:Using hybrid capture-chemiluminescence HPV tests for HPV 16/18 genotyping, combined with the viral loads (>10 RLU/CO) of the other 12 high-risk HPV analysis, one could triage HPV positive population without additional tests. Such triage strategy could promote the coverage of cervical cancer screening, particularly where cytology pathologists or economic resources are limited.

Objective:To evaluate the clinical value of high-risk human papillomavirus (HPV) viral load for the cervical cancer screening and triage of high-risk HPV positive populations without additional tests.Methods:(1) This study conducted a retrospective analysis of 29 720 women aged 35-64 years who received cervical cancer screening in Quanzhou, China, in 2021. Fourteen high-risk HPV types (including HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) were detected for cervical cancer primary screening using hybrid capture-chemiluminescence method. High-risk HPV positive samples were further subjected to HPV 16/18 genotyping using hybrid capture-chemiluminescence method. Among them, HPV 16/18 positive women were directly referred to colposcopy, while the other 12 high-risk HPV positive samples were further subjected to liquid based cytology test. Those with abnormal or suspicious cytology were referred to colposcopy. Biopsies were taken for histopathological examination of suspicious or abnormal individuals under colposcopy. (2) Ten cases of colposcopy loss or refusal to undergo examination were excluded, and the data from the 29 710 cases were analyzed. The HPV viral loads of the other 12 high-risk HPV positive populations were focused and evaluated their HPV viral loads for further cervical intraepithelial neoplasia (CIN) Ⅱ and above lesions (CINⅡ +) triage in cervical cancer screening. Results:(1) Among 29 720 women, 2 487 women (8.37%, 2 487/29 720) were positive for high-risk HPV, including 807 women (2.72%, 807/29 720) were positive for HPV 16/18 and 1 680 patients (5.65%, 1 680/29 720) were positive for the other 12 high-risk HPV types. Among 1 680 women who tested positive for the other 12 high-risk HPV types, 573 patients were atypical squamous cell carcinoma of unclear significance or above, 346 patients were CIN Ⅰ, 122 patients were CIN Ⅱ-Ⅲ, 9 patients were squamous cell carcinoma patients, and 4 patients were adenocarcinoma in situ. The immediate risk of CIN Ⅱ + in HPV 16/18 positive women (11.13%) was approximately four times higher than that of other 12 high-risk HPV positive women (2.74%). (2) Through the viral load analysis of the other 12 high-risk HPV types, we found that the viral load of the other 12 high-risk HPV provide a good value for the pathological results, with a clinical cutoff (CO) value of 11.21 relative light unit/CO (RLU/CO) for the CINⅡ + detection. Except for HPV 16/18 positive patients, when the viral load values of the other 12 high-risk HPV types were greater than 10 RLU/CO, these patients had a higher risk of CINⅡ +, with a positive predictive value of 31.29%. CINⅡ + was not found in any of the other 12 high-risk HPV positive with viral load values less than or equal to 10 RLU/CO. Conclusions:Using hybrid capture-chemiluminescence HPV tests for HPV 16/18 genotyping, combined with the viral loads (>10 RLU/CO) of the other 12 high-risk HPV analysis, one could triage HPV positive population without additional tests. Such triage strategy could promote the coverage of cervical cancer screening, particularly where cytology pathologists or economic resources are limited.

Objective To explore the feasibility, safety, and efficacy of transesophageal endoscopic surgery for mediastinal tumors. Methods A retrospective analysis was conducted on the clinical data of 17 patients who underwent transesophageal endoscopic resection for benign mediastinal tumors at the Endoscopy Center of Zhongshan Hospital, Fudan University, between January 1, 2016 and December 31, 2024. Epidemiological characteristics, surgical parameters, adverse events, and follow-up outcomes were analyzed. Results Among the 17 patients, there were 9 males and 8 females, with an average age of (42.4±14.5) years and an average tumor size of (2.6±1.6) cm. Pathological types included esophageal duplication cysts (6 cases, 35.3%), bronchogenic cysts (5 cases, 29.4%), gastroenteric cysts (3 cases, 17.6%), schwannomas (2 cases, 11.8%), and lymphangioma (1 case, 5.9%). Fourteen patients (82.4%) underwent submucosal tunneling endoscopic resection (STER), 3 patients (17.6%) underwent natural orifice transluminal endoscopic mediastinal surgery. All surgeries were successfully completed without conversion to open surgery. En bloc resection was achieved in 11 patients (64.7%), with an average operative time of (60.9±32.6) min. No intraoperative bleeding or mucosal injury occurred, and 4 patients (23.5%) experienced minor complications (pneumothorax, fever, recurrent laryngeal nerve injury), all of which resolved with conservative treatment. The average postoperative hospital stay was (3.2±1.5) days, and no recurrence was observed during the follow-up period. Conclusions Transesophageal endoscopic resection of benign mediastinal tumors is a safe, effective, and minimally invasive treatment method. Further validation of its efficacy and safety through large-scale prospective studies is warranted.

Objective:To evaluate the efficacy of cyclophosphamide in patients with T-cell large granular lymphocytic leukemia (T-LGLL) and the maintenance of treatment-free remission (TFR) following drug discontinuation.Methods:Clinical data were collected from 37 patients with T-LGLL who received oral cyclophosphamide at the Regenerative Medicine Clinic of the Institute of Hematology and Blood Diseases Hospital between June 2019 and March 2024. Patient clinical characteristics, treatment efficacy, and long-term TFR were analyzed.Results:The median age of the 37 patients was 60 years (range: 37-86), and 22 (59.5%) were male. Anemia was observed in 30 patients (81.1%), and 28 (75.7%) met the diagnostic criteria for secondary pure red cell aplasia. Neutropenia occurred in 15 patients (40.5%), lymphocytosis in 11 (29.7%), and thrombocytopenia in three (8.1%). Sixteen patients (43.2%) had not received prior immunosuppressive therapy (treatment-naive group), while 21 patients (56.8%) were refractory to or had relapsed after immunosuppressive treatment (refractory/relapsed group). All patients met the treatment criteria and received oral cyclophosphamide at doses of 50-100 mg/day. Among the 36 evaluable patients, hematologic remission was achieved in 25 (69.4%), with a median time of 2.0 months (range: 0.7-7.0). There was no statistically significant difference in remission rates between the treatment-naive and refractory/relapsed groups (68.5% vs. 66.7%, P=0.589). Among the 25 patients who achieved hematologic remission, 24 discontinued cyclophosphamide. With a median follow-up of 39.0 months (range: 8.0-56.0), the median TFR duration was not reached. The estimated TFR rates were (90.87± 6.16) % at 12 months and (75.72±11.04) % at 36 months. No significant difference in TFR was observed between the treatment-naive and refractory/relapsed groups ( P=0.451) . Conclusion:Oral cyclophosphamide is effective in the treatment of T-LGLL, and patients may maintain long-term TFR following drug discontinuation.

Objective To investigate the effects of indirect moxibustion on the expressions of DNA methyltransferases(DNMT)and methylation of the brain-derived neurotrophic factor(BDNF)promoter region in uterine tissues of rats with primary dysmenorrhea(PD);To explore the mechanism of epigenetic regulation of indirect moxibustion on PD model rats.Methods A total of 32 female SD rats were randomly divided into blank group,model group,indirect moxibustion group and Western medicine group,with 8 rats in each group.The PD model with cold dampness stagnation syndrome was established using ice-water baths combined with estradiol benzoate and oxytocin.Starting from the first day of modeling,the indirect moxibustion group received salt-partitioned moxibustion at"Shenque"and ginger-partitioned moxibustion at"Guanyuan"for 20 min,while the Western medicine group was gavaged ibuprofen solution.Both interventions were given once a day for 10 days.On day 11,writhing responses were observed and scored after oxytocin injection,Western blot and RT-qPCR were used to detect protein and mRNA expression of BDNF,DNMT3A and DNMT3B in uterine tissue,immunohistochemical staining was used to detect the positive expressions of DNMT3A and DNMT3B in uterine tissue.The DNA methylation of BDNF promoter region in uterine tissue was detected by sulfite sequencing.Results Compared with the blank group,the writhing latency was shortened and the writhing score increased in the model group(P<0.01);the protein and mRNA expressions of BDNF,DNMT3A and DNMT3B in uterine tissue increased(P<0.01),the positive expressions of DNMT3A and DNMT3B increased(P<0.01),and the DNA methylation rate in BDNF promoter region decreased(P<0.01).Compared with the model group,the writhing latency was lengthened and the writhing score decreased in the indirect moxibustion group and Western medicine group(P<0.05,P<0.01);the protein and mRNA expressions of BDNF,DNMT3A and DNMT3B in uterine tissue decreased(P<0.05,P<0.01),the positive expressions of DNMT3A and DNMT3B decreased(P<0.01),and the DNA methylation rate in BDNF promoter region increased(P<0.01).Conclusion Indirect moxibustion at"Shenque"and"Guanyuan"may inhibit the transcription of BDNF by increasing the DNA methylation level of BDNF promoter region,and reduce the expression of BDNF,so as to relieve the pain of PD rats.

Objective:To identify factors influencing orthostatic intolerance (OI) in patients during initial ambulation following minimally invasive lung surgery and develop and validate a risk prediction model to assist clinical practitioners in screening high-risk patients.Methods:Totally 1 000 patients who underwent minimally invasive lung surgery at the Department of Thoracic Surgery of Peking University Third Hospital from March 2022 to November 2023 were recruited by convenience sampling. Patients were randomly divided into a modeling group ( n=800) and an internal validation group ( n=200) in an 8∶2 ratio. Univariate analysis and logistic regression were applied to determine risk factors for OI in the modeling group. R software was utilized to construct a nomogram model. The model's predictive performance was assessed using the area under the ROC curve ( AUC) for both the modeling and validation groups. Calibration curves were plotted to evaluate consistency, and the Hosmer-Lemeshow test was conducted to confirm model fit. Results:The incidence of OI during initial ambulation was 37.2% (372/1 000). Logistic regression identified BMI, postoperative day 1 drainage volume, postoperative use of nonsteroidal anti-inflammatory drugs (NSAIDs), and initial ambulation pain score as independent risk factors for OI ( P<0.05). The AUC for the nomogram model in the modeling group was 0.645, and 0.694 in the validation group, indicating good predictive accuracy. Calibration curves showed strong agreement between predicted and observed outcomes ( P>0.05) . Conclusions:The constructed risk prediction model demonstrates good predictive ability for OI risk during initial ambulation following minimally invasive lung surgery, which can support clinical identification of high-risk patients. This tool may provide valuable guidance for implementing early, targeted preventive measures.

Objective The long-term epidemiological and etiological characteristics of hand, foot and mouth disease (HFMD) in Huai’an, Jiangsu were analyzed to provide scientific evidence for the prevention of HFMD. Methods The data of HFMD reports, etiological diagnosis and in Huai’an from 2009 to 2022 were described and analyzed. Results A total of 78 535 cases were reported from 2009 to 2022 , with 14-year average annual incidence rate of 114.71/100 000. Before 2020, the incidence rate of HFMD in Huai’an showed the epidemic intensity increased every other year on the whole, and the average annual incidence rate during the COVID-19 pandemic (2020-2022) (55.69/100 000) was significantly lower than that in previous years (2009-2019) (129.95/100 000). The joinpoint regression analysis showed that the best fitting model from 2009 to 2022 had no joinpoints, APC=AAPC=-1.24%. The overall trend showed a monotonously decreasing trend, but the trend was not statistically significant. The male-to-female distribution ratio was 1.53:1, and the age distribution was mainly under 5 years old, especially in scattered children. The epidemic season was from April to July. The results of etiological surveillance showed that the co-epidemic of Coxsackievirus A16 (CV-A16) and Enterovirus A71 (EV-A71) during the early stages had changed to the co-epidemic of CV-A16 and CV-A6 in the current period. Conclusion The burden of HFMD in Huai’an was large, and the epidemic intensity increased every other year was affected by the COVID-19 pandemic. The epidemiological features after the COVID-19 pandemic should be further monitored.

Modulating Tankyrases (TNKS), interactions with USP25 to promote TNKS degradation, rather than inhibiting their enzymatic activities, is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway. Here, we identified UAT-B, a novel neoantimycin analog isolated from Streptomyces conglobatus, as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction (PPI) to overcome multi-drug resistance in colorectal cancer (CRC). The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels, triggering cell apoptosis through modulation of the Wnt/β-catenin pathway. Importantly, UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels, as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts, as well as APC^min/+ spontaneous CRC models. Collectively, these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment, and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.