The ethical review of clinical research on rare diseases is crucial in ensuring the scientific validity of the research and the rights and interests of the subjects. Starting from the definition of rare diseases， this paper analyzed the current situation of domestic and international regulations and ethical review in clinical research on rare diseases. It also explored the key elements of ethical review from the two dimensions of scientific and ethical aspects of clinical research， including research objectives， methods， risk and benefit assessment， researcher qualifications， research infrastructure， informed consent process， data security and privacy protection， and protection of vulnerable groups such as children. Regarding the ethical review of clinical research on rare diseases， strategies can be adopted such as strengthening the training of ethics review personnel， conducting multi-center collaborative reviews， and focusing on the long-term safety of trials， to improve the quality of ethical review， protect the safety of the subjects， and ensure the efficiency and quality of clinical research.

Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective:To explore the configurational factors influencing the professional commitment of Master of Nursing Specialist (MNS) students and identify effective paths to enhance their professional commitment.Methods:A convenience sample of MNS students from six hospitals in Jiangsu Province, Shanghai City, and Anhui Province was surveyed between December 2023 and October 2024. Data were collected using a general information questionnaire, the Nurses' Professional Commitment Scale (NPCS), the Connor-Davidson Resilience Scale (CD-RISC-10), the Multidimentional Perceived Social Support (MSPSS), and the Vocational Outcome Expectation Scale (VOE-R). Fuzzy-set Qualitative Comparative Analysis (fsQCA) was used to explore the configurational influences on professional commitment.Results:A total of 332 questionnaires were distributed, and 302 valid responses were obtained (effective response rate: 90.1%). The mean NPCS score among respondents was (89.30±22.42), indicating a upper-middle level of professional commitment. The fsQCA results revealed that the frequency of night shift rotation was a necessary condition affecting professional commitment. A total of three distinct configurations (paths) were identified that can enhance the professional commitment of MNS students.Conclusions:The professional commitment of MNS students is influenced by multiple interacting factors. It is recommended that hospitals, universities, and nursing educators develop targeted interventions based on these identified configurations to improve professional commitment.

Objective:To explore the configurational factors influencing the professional commitment of Master of Nursing Specialist (MNS) students and identify effective paths to enhance their professional commitment.Methods:A convenience sample of MNS students from six hospitals in Jiangsu Province, Shanghai City, and Anhui Province was surveyed between December 2023 and October 2024. Data were collected using a general information questionnaire, the Nurses' Professional Commitment Scale (NPCS), the Connor-Davidson Resilience Scale (CD-RISC-10), the Multidimentional Perceived Social Support (MSPSS), and the Vocational Outcome Expectation Scale (VOE-R). Fuzzy-set Qualitative Comparative Analysis (fsQCA) was used to explore the configurational influences on professional commitment.Results:A total of 332 questionnaires were distributed, and 302 valid responses were obtained (effective response rate: 90.1%). The mean NPCS score among respondents was (89.30±22.42), indicating a upper-middle level of professional commitment. The fsQCA results revealed that the frequency of night shift rotation was a necessary condition affecting professional commitment. A total of three distinct configurations (paths) were identified that can enhance the professional commitment of MNS students.Conclusions:The professional commitment of MNS students is influenced by multiple interacting factors. It is recommended that hospitals, universities, and nursing educators develop targeted interventions based on these identified configurations to improve professional commitment.

Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective To explore the effects of Huoxue Jiedu Formula on the expression of the apoptosis-related factors B-cell lymphoma-2(Bcl-2)and survivin in submaxillary gland cells in mice with Sj?gren syndrome(SS).Methods Using 15 healthy C57BL/6J female mice as the blank control group.Seventy-five of non-obese diabetes mellitus female mice were classified as SS model.They were randomly assigned to the model group,total glucosides of paeony capsule group(TGP group,0.233 g/kg),Huoxue Jiedu Formula low-(15.925 g/kg),middle-(31.850 g/kg),and high-dose(62.335 g/kg)groups,with 15 mice in each group.The blank control and model groups received distilled water by gavage,and the other groups received respective drug doses by gavage once a day for 8 consecutive weeks.After 1,2,4,6,and 8 weeks of gavage,the salivary secretion volume of the mice was measured.The submaxillary gland index was calculated.Pathological changes in the submaxillary gland of mice were observed by hematoxylin-eosin staining.The expression of apoptosis-related factors Bcl-2 and survivin was detected by Western blotting and real-time fluorescence PCR.Results There was statistical significance in salivary secretion volume from aspects of between-group,time-point,and interaction effects(P<0.05).After 8 weeks of treatment,compared to the blank control group,salivary secretion volume,submaxillary gland index,and protein and mRNA expressions of Bcl-2 and survivin in the model group were decreased,while the pathological score grading increased(P<0.05).Compared to the model group,the TGP group and the Huoxue Jiedu Formula low-,medium-,and high-dose groups showed an increase in saliva secretion after 8 weeks of treatment and an increase in submaxillary gland index,Bcl-2,and survivin mRNA expression levels,and a decrease in pathological score levels.The protein expressions of Bcl-2 and survivin in the Huoxue Jiedu Formula medium-and high-dose groups were increased(P<0.05).Compared to the TGP group,salivary secretion volume and submaxillary gland index in the Huoxue Jiedu Formula low-dose group were lower at weeks 4 and 6,while the pathological score grading was higher(P<0.05).The pathological score grading in the Huoxue Jiedu Formula high-dose group was lower.The protein and mRNA expressions of Bcl-2 in the Huoxue Jiedu Formula low-and medium-dose groups were lower(P<0.05).Conclusion Huoxue Jiedu Formula may relieve the submaxillary gland lesions and increase the salivary secretion volume of the submaxillary gland in SS model mice by regulating the expressions of apoptosis-related factors such as Bcl-2 and survivin in the submaxillary gland.

Objective To explore the effects of Huoxue Jiedu Formula on the expression of the apoptosis-related factors B-cell lymphoma-2(Bcl-2)and survivin in submaxillary gland cells in mice with Sj?gren syndrome(SS).Methods Using 15 healthy C57BL/6J female mice as the blank control group.Seventy-five of non-obese diabetes mellitus female mice were classified as SS model.They were randomly assigned to the model group,total glucosides of paeony capsule group(TGP group,0.233 g/kg),Huoxue Jiedu Formula low-(15.925 g/kg),middle-(31.850 g/kg),and high-dose(62.335 g/kg)groups,with 15 mice in each group.The blank control and model groups received distilled water by gavage,and the other groups received respective drug doses by gavage once a day for 8 consecutive weeks.After 1,2,4,6,and 8 weeks of gavage,the salivary secretion volume of the mice was measured.The submaxillary gland index was calculated.Pathological changes in the submaxillary gland of mice were observed by hematoxylin-eosin staining.The expression of apoptosis-related factors Bcl-2 and survivin was detected by Western blotting and real-time fluorescence PCR.Results There was statistical significance in salivary secretion volume from aspects of between-group,time-point,and interaction effects(P<0.05).After 8 weeks of treatment,compared to the blank control group,salivary secretion volume,submaxillary gland index,and protein and mRNA expressions of Bcl-2 and survivin in the model group were decreased,while the pathological score grading increased(P<0.05).Compared to the model group,the TGP group and the Huoxue Jiedu Formula low-,medium-,and high-dose groups showed an increase in saliva secretion after 8 weeks of treatment and an increase in submaxillary gland index,Bcl-2,and survivin mRNA expression levels,and a decrease in pathological score levels.The protein expressions of Bcl-2 and survivin in the Huoxue Jiedu Formula medium-and high-dose groups were increased(P<0.05).Compared to the TGP group,salivary secretion volume and submaxillary gland index in the Huoxue Jiedu Formula low-dose group were lower at weeks 4 and 6,while the pathological score grading was higher(P<0.05).The pathological score grading in the Huoxue Jiedu Formula high-dose group was lower.The protein and mRNA expressions of Bcl-2 in the Huoxue Jiedu Formula low-and medium-dose groups were lower(P<0.05).Conclusion Huoxue Jiedu Formula may relieve the submaxillary gland lesions and increase the salivary secretion volume of the submaxillary gland in SS model mice by regulating the expressions of apoptosis-related factors such as Bcl-2 and survivin in the submaxillary gland.

Objective:To explore the therapeutic characteristics of population with gout achieving treat-to-target (T2T) indicators through real-world research and evaluate their safety.Methods:A total of 3 287 patients diagnosed with gout by rheumatologists in 21 first-class tertiary hospitals in 10 provinces, municipalities, and autonomous regions in China from January 2015 to December 2021 were included in this polycentric cross-sectional study. The database included patients′ general information, disease characteristics, and clinical application of traditional Chinese and Western medicine treatment measures. SPSS and Excel software were used for data analysis. Frequency analysis, cluster analysis, and factor analysis were used to summarize the characteristics and rules of treatment measures for patients with gout who achieved the target after treatment. The occurrence of adverse events (AE) was recorded during treatment.Results:After treatment, 691 visits (7%) achieved the serum urate (SUA) target, and the most frequent use of urate-lowering therapy (ULT) was febuxostat, followed by benzbromarone. The most common treatment options were following: GroupⅠ: traditional Chinese medicine (TCM) decoction-TCM external treatment-physical exercise-proprietary Chinese medicine; GroupⅡ: ferulic acid-nonsteroidal anti-inflammatory drugs (NSAIDs); Group Ⅲ: allopurinol-sodium bicarbonate-benzbromarone; Group Ⅳ: glucocorticoid-colchicine; Group Ⅴ: febuxostat. A total of 5 898 visits (60%) chieved manifestations of joint pain VAS scores target, and the most frequently used drug to control joint symptoms was NSAIDs. The frequency of use of drugs to control joint symptoms were 2 118 times (usage rate reached 35.9%), while the frequency of ULT were 2 504 times (usage rate reached 42.5%), which was higher than the joint symptom control drug. The most common treatment options were following: Group Ⅰ: proprietary Chinese medicine-TCM decoction-TCM external treatment-physical exercise; Group Ⅱ: NSAIDs-colchicine hormones; Group Ⅲ: allopurinol, Group Ⅳ: benzbromarone; Group Ⅴ: febuxostat. A total of 59 adverse events occurred during treatment.Conclusion:The proportions of gout patients who reach target serum urate level & good control of joint symptoms are both very low, and ULT and anti-inflammatory prescription patterns are very different from international guidelines, so it is necessary to strengthen the standardized management of gout patients. At the same time, life intervention measures account for a certain proportion of the treatment plans for the T2T population, and further exploration is needed.

Listeria monocytogenes (Lm) is zoonotic pathogen that can cause listeriosis, and vaccine is one of the effective methods to prevent this pathogen infection. In this study, we developed a novel vaccine that is a mixture of inactivated bacteria and Montanide™ ISA 61 VG, a mineral oil adjuvant, and evaluated the safety and immune response characteristics of this vaccine. The mice immunized with the ISA 61 VG adjuvant had high safety, and it could induce significantly higher titer of anti-listeriolysin O (LLO) antibody and higher value of IgG2a/IgG1 ratio compared with the group without the adjuvant. In particular, it could provide 100% immune protection against lethal doses of Lm challenge in mice. In summary, ISA 61VG adjuvant significantly enhanced the ability of inactivated listeria vaccine to induce humoral and cellular immune responses, thereby enhanced the protective immune response in the host, and it is a potential vaccine candidate for the prevention of Lm infection in humans and animals.
Adjuvants, Immunologic ; pharmacology ; Animals ; Hemolysin Proteins ; immunology ; pharmacology ; Immunity, Cellular ; drug effects ; Listeria monocytogenes ; immunology ; Listeriosis ; prevention & control ; Mice ; Mice, Inbred BALB C ; Vaccines, Inactivated ; immunology

Objective To investigate the curative effect of bronchoalveolar lavage with fiberoptic bronchoscopy combined with vibration sputum drainage in the treatment of severe pneumonia patients undergoing mechanical ventilation (MV).Methods A prospective randomized controlled trial was conducted. 286 severe pneumonia patients undergoing MV admitted to intensive care unit (ICU) of Hunan People's Hospital from January 2014 to July 2016 were enrolled, and they were divided into control group and observation group according to random number table, with 143 patients in each group. Patients in both groups received sensitive antibiotics for anti-infection, etiological treatment, and calefacient and humidifying treatment. The patients in the control group received bronchoalveolar lavage with fiberoptic bronchoscopy, and those in the observation group received bronchoalveolar lavage combined with vibration sputum drainage. The parameters of respiratory function and inflammation before and after treatment, curative effect, and prognosis were compared between the two groups.Results ① There were no significant differences in respiratory function parameters between the two groups before treatment, 2 hours after treatment, the parameters were improved in both groups. Moreover, oxygenation index (PaO2/FiO2) in observation group was significantly higher than that of control group [mmHg (1 mmHg = 0.133 kPa): 379.1±20.2 vs. 351.8±24.7], and arterial partial pressure of carbon dioxide (PaCO2) and airway resistance (Raw) were significantly lower than those of the control group[PaCO2 (mmHg): 36.5±5.8 vs. 45.3±6.9, Raw (cmH2O, 1 cmH2O = 0.098 kPa): 12.9±0.6 vs. 13.1±0.8, allP < 0.01]. ② There were no significant differences in inflammation parameters between the two groups before treatment, 24 hours after intervention, which were significantly decreased in both groups. Moreover, white blood cell count (WBC), procalcitonin (PCT) and C-reactive protein (CRP) in the observation group were significantly lower than those of the control group [WBC (×109/L): 8.2±1.7 vs. 12.8±3.7, PCT (μg/L): 15.4±2.4 vs. 21.8±3.1, CRP (mg/L): 37.1±6.1 vs. 67.2±7.2, allP < 0.01]. ③ Compared with the control group, the treatment efficiency of observation group was improved [95.1% (136/143) vs. 87.4% (125/143)], the quantity of sputum excretion was increased (mL: 49.2±12.5 vs. 36.9±11.0), duration of MV and length of ICU stay were significantly shortened (days: 6.4±3.6 vs. 9.4±2.1, 8.6±5.7 vs. 12.4±4.6, bothP < 0.01), however, there was no significantly statistical difference in 28-day mortality between control group and observation group [2.8% (4/143) vs. 2.1% (3/143),P > 0.05].Conclusion Compared with bronchoalveolar lavage with fiberoptic bronchoscopy alone, the treatment of bronchoalveolar lavage combined with vibration sputum drainage is more effective in sputum excretion for severe pneumonia patients undergoing MV, which could improve the respiratory function, reduce infection, shorten the duration of MV and the length of ICU stay, and improve the recovery.