1.Analysis of epigenetic characteristics in colonic tumors of Apcmin/+via spatial ATAC-seq technology
Lebin LIANG ; Huifang CHEN ; Shujing LAI ; Liang GU ; Bing SU
Journal of Shanghai Jiaotong University(Medical Science) 2025;45(10):1261-1270
Objective·To investigate the spatial epigenetic characteristics of spontaneous colon tumors in Apcmin/+mice.Methods·A spatial assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)technology platform was established using an eight-month-old male Apcmin/+mouse model with spontaneous colon tumors.One tumor from a mouse was harvested and embedded in OCT compound for serial cryosectioning;one tissue section was stained with hematoxylin-eosin(H-E)to observe its histological characteristics,while an adjacent section was processed using spatial ATAC-seq technology to generate spatially resolved DNA libraries,followed by sequencing to obtain spatial chromatin accessibility data.Another tumor from the same mouse was digested into a single-cell suspension,in which viable single cells were sorted by flow cytometry and processed for single-cell RNA sequencing.The results were integrated with spatial chromatin accessibility data to jointly analyze the epigenetic characteristics of the colon tumor microenvironment.Results·A stable spatial ATAC-seq platform was successfully established,dividing the tumor into malignant,non-malignant,and malignant-non-malignant boundary regions.Transcription factors enriched in malignant regions included NK2 homeobox 5(NKX2-5)and transcription factor 3(TCF3).Analysis of transcription factor enrichment in the 3 regions revealed two distinct expression trends:one showing a gradual decrease from malignant to boundary to non-malignant regions,and the other exhibiting high expression in malignant and boundary regions but low expression in non-malignant regions.Gene analysis across regions revealed significant upregulation of hypoxia response,transforming growth factor(TGF),and Kirsten rat sarcoma viral oncogene homolog(KRAS)signaling pathways in malignant regions,with cell cycle-related functions markedly enhanced.Analysis of cell-cell interactions in the tumor microenvironment revealed significant differences in interaction strength:strong interactions within non-malignant regions,moderate interactions between boundary and non-malignant regions,and weak interactions between malignant and boundary regions as well as between malignant and non-malignant regions.Conclusion·Colon tumors in Apcmin/+mice exhibit high spatial heterogeneity;malignant regions were enriched with transcription factors including TCF3,and cell interactions between malignant regions and boundary/non-malignant regions were relatively weak.
2.Analysis of epigenetic characteristics in colonic tumors of Apcmin/+via spatial ATAC-seq technology
Lebin LIANG ; Huifang CHEN ; Shujing LAI ; Liang GU ; Bing SU
Journal of Shanghai Jiaotong University(Medical Science) 2025;45(10):1261-1270
Objective·To investigate the spatial epigenetic characteristics of spontaneous colon tumors in Apcmin/+mice.Methods·A spatial assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)technology platform was established using an eight-month-old male Apcmin/+mouse model with spontaneous colon tumors.One tumor from a mouse was harvested and embedded in OCT compound for serial cryosectioning;one tissue section was stained with hematoxylin-eosin(H-E)to observe its histological characteristics,while an adjacent section was processed using spatial ATAC-seq technology to generate spatially resolved DNA libraries,followed by sequencing to obtain spatial chromatin accessibility data.Another tumor from the same mouse was digested into a single-cell suspension,in which viable single cells were sorted by flow cytometry and processed for single-cell RNA sequencing.The results were integrated with spatial chromatin accessibility data to jointly analyze the epigenetic characteristics of the colon tumor microenvironment.Results·A stable spatial ATAC-seq platform was successfully established,dividing the tumor into malignant,non-malignant,and malignant-non-malignant boundary regions.Transcription factors enriched in malignant regions included NK2 homeobox 5(NKX2-5)and transcription factor 3(TCF3).Analysis of transcription factor enrichment in the 3 regions revealed two distinct expression trends:one showing a gradual decrease from malignant to boundary to non-malignant regions,and the other exhibiting high expression in malignant and boundary regions but low expression in non-malignant regions.Gene analysis across regions revealed significant upregulation of hypoxia response,transforming growth factor(TGF),and Kirsten rat sarcoma viral oncogene homolog(KRAS)signaling pathways in malignant regions,with cell cycle-related functions markedly enhanced.Analysis of cell-cell interactions in the tumor microenvironment revealed significant differences in interaction strength:strong interactions within non-malignant regions,moderate interactions between boundary and non-malignant regions,and weak interactions between malignant and boundary regions as well as between malignant and non-malignant regions.Conclusion·Colon tumors in Apcmin/+mice exhibit high spatial heterogeneity;malignant regions were enriched with transcription factors including TCF3,and cell interactions between malignant regions and boundary/non-malignant regions were relatively weak.
3.Value of preoperative salivary cortisol morning/night secretion ratio in predicting acute postoperative pain
Shujing GU ; Cunjin WANG ; Ju GAO ; Huaizhong MO
Chinese Journal of Anesthesiology 2024;44(8):917-921
Objective:To assess the value of preoperative salivary cortisol morning/night secretion ratio (AM/PM ratio) in predicting acute postoperative pain in patients.Methods:This was a prospective cohort study. A total of 120 patients, aged 18-64 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, with body mass index of 18-28 kg/m 2, undergoing elective single-port laparoscopic hysterectomy from January to October 2023 in Subei People′s Hospital of Jiangsu Province, were selected, and their anxiety and depression status was evaluated by using the Hospital Anxiety and Depression Mood Measurement Scale (HADS) before surgery. Saliva samples were collected in the morning and evening 1 day before operation, and the cortisol concentration was determined by enzyme-linked immunosorbent assay. The ratio of morning cortisol concentration to night cortisol concentration before operation was calculated (cortisol AM/PM ratio). The patients were divided into two groups according to the highest numerical rating scale (NRS) score within 48 h after surgery: no or mild pain group with the highest NRS score ≤3 and moderate to severe pain group with the highest NRS score ≥ 4. Multivariate logistic regression was used to identify the risk factors of moderate to severe acute pain after surgery, and the receiver operating characteristic curve was plotted to evaluate the predictive value. Results:Multivariate logistic regression analysis showed that age, preoperative HADS anxiety score, preoperative HADS depression score and preoperative cortisol AM/PM ratio were independent risk factors for moderate to severe acute pain after surgery ( P<0.05). The results of the receiver operating characteristic curve showed that the area under the curve of preoperative cortisol AM/PM ratio in predicting moderate to severe acute postoperative pain was 0.803 (95% confidence interval 0.708-0.898, P<0.001), the optimal cut-off value was 3.46, the sensitivity was 71.1%, and the specificity was 87.7%. Conclusions:Preoperative salivary cortisol morning/night secretion ratio can predict the occurrence of acute postoperative pain in patients.
4.A qualitative study on disorienting dilemmas of master of nursing specialist
Zhijia SHEN ; Caifeng LUO ; Jian'ou XU ; Shujing GU ; Fei LYU ; Dongmei ZHU ; Xiaoyun GUO
Chinese Journal of Modern Nursing 2020;26(27):3744-3750
Objective:To explore disorienting dilemmas during postgraduate study period experienced by postgraduates with master of nursing specialist who participated in the national postgraduate entrance examination and provide a basis for teaching reform based on transformative learning theory.Methods:Using the purposive sampling method and the maximum difference method, 23 full-time masters of nursing specialist who participated in the national postgraduate entrance examination from 3 universities of Jiangsu Province were selected as the research objects from March to June 2019. The descriptive qualitative research method was used to conduct semi-structured interviews, and the traditional content analysis method was used to analyze the interview data.Results:Four themes of disorienting dilemmas of masters of nursing specialist were extracted: the expected hypothesis of challenges of new environment in course learning, a gap between scientific research ability and training target, adjustment of self-positioning in clinical practice and insufficient preparations for graduation of master of nursing specialist.Conclusions:Master of nursing specialist have disorienting dilemmas in course study, scientific research, clinical practice and graduation preparation, and nursing educators can use this as a trigger point to carry out teaching reform based on transformative learning theory.

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