1.Application of CRISPR/Cas System in Precision Medicine for Triple-negative Breast Cancer
Hui-Ling LIN ; Yu-Xin OUYANG ; Wan-Ying TANG ; Mi HU ; Mao PENG ; Ping-Ping HE ; Xin-Ping OUYANG
Progress in Biochemistry and Biophysics 2025;52(2):279-289
Triple-negative breast cancer (TNBC) represents a distinctive subtype, characterized by the absence of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor 2 (HER2). Due to its high inter-tumor and intra-tumor heterogeneity, TNBC poses significant chanllenges for personalized diagnosis and treatment. The advant of clustered regular interspaced short palindromic repeats (CRISPR) technology has profoundly enhanced our understanding of the structure and function of the TNBC genome, providing a powerful tool for investigating the occurrence and development of diseases. This review focuses on the application of CRISPR/Cas technology in the personalized diagnosis and treatment of TNBC. We begin by discussing the unique attributes of TNBC and the limitations of current diagnostic and treatment approaches: conventional diagnostic methods provide limited insights into TNBC, while traditional chemotherapy drugs are often associated with low efficacy and severe side effects. The CRISPR/Cas system, which activates Cas enzymes through complementary guide RNAs (gRNAs) to selectively degrade specific nucleic acids, has emerged as a robust tool for TNBC research. This technology enables precise gene editing, allowing for a deeper understanding of TNBC heterogeneity by marking and tracking diverse cell clones. Additionally, CRISPR facilitates high-throughput screening to promptly identify genes involved in TNBC growth, metastasis, and drug resistance, thus revealing new therapeutic targets and strategies. In TNBC diagnostics, CRISPR/Cas was applied to develop molecular diagnostic systems based on Cas9, Cas12, and Cas13, each employing distinct detection principles. These systems can sensitively and specifically detect a variety of TNBC biomarkers, including cell-specific DNA/RNA and circulating tumor DNA (ctDNA). In the realm of precision therapy, CRISPR/Cas has been utilized to identify key genes implicated in TNBC progression and treatment resistance. CRISPR-based screening has uncovered potential therapeutic targets, while its gene-editing capabilities have facilitated the development of combination therapies with traditional chemotherapy drugs, enhancing their efficacy. Despite its promise, the clinical translation of CRISPR/Cas technology remains in its early stages. Several clinical trials are underway to assess its safety and efficacy in the treatment of various genetic diseases and cancers. Challenges such as off-target effects, editing efficiency, and delivery methods remain to be addressed. The integration of CRISPR/Cas with other technologies, such as 3D cell culture systems, human induced pluripotent stem cells (hiPSCs), and artificial intelligence (AI), is expected to further advance precision medicine for TNBC. These technological convergences can offer deeper insights into disease mechanisms and facilitate the development of personalized treatment strategies. In conclusion, the CRISPR/Cas system holds immense potential in the precise diagnosis and treatment of TNBC. As the technology progresses and becomes more costs-effective, its clinical relevance will grow, and the translation of CRISPR/Cas system data into clinical applications will pave the way for optimal diagnosis and treatment strategies for TNBC patients. However, technical hurdles and ethical considerations require ongoing research and regulation to ensure safety and efficacy.
2.Protective effects of exosomes derived from MSCs in radiation-induced lung injury
Lili WANG ; Zien YANG ; Mingyue OUYANG ; Sining XING ; Song ZHAO ; Huiying YU
Chinese Journal of Radiological Health 2025;34(1):13-20
Objective To investigate the role and related mechanisms of exosomes derived from mesenchymal stem cells (MSCs) in radiation-induced lung injury (RILI). Methods Human umbilical cord-derived MSCs were isolated and cultured for the extraction and identification of exosomes. Eighteen male SD rats were randomly divided into Control group, RILI group and RILI + exosomes group (EXO group), with 6 rats in each group. Except for Control group, the other groups received a single X-ray dose of 30 Gy to the right lung. Immediately after irradiation, the EXO group was administered 2 × 109 exosomes/kg via tail vein injection. Control group and RILI group were given the same volume of normal saline. Eight weeks post-irradiation, the rats were sacrificed, lung tissue and peripheral venous blood were collected. HE and Masson staining were employed to observe the pathological and fibrotic changes of lung tissue. The levels of serum inflammatory factors IL-6, IFN-γ, TNF-α, and IL-10 were detected by ELISA. RT-qPCR was used to assess the mRNA levels of IL-1β, IL-6, Cdh1, and Col1a1 in lung tissue. The expression levels of Vimentin and TGF-β1 in lung tissue were measured by immunohistochemical staining. The expression levels of AMPK, p-AMPK, and TGF-β1 in lung tissue were detected by Western blot. Results MSC-derived exosomes were successfully extracted and identified. Compared with RILI group, EXO group showed significantly reduced pathological changes of lung inflammation and collagen deposition. The levels of serum inflammatory factors IL-6, INF-γ, and TNF-α were significantly decreased (P < 0.05), and the level of anti-inflammatory factor IL-10 was significantly increased (P < 0.05). The mRNA levels of IL-1β, IL-6, and Col1a1 in lung tissue were significantly decreased (P < 0.05 or P < 0.01), and the mRNA level of Cdh1 was significantly increased (P < 0.05 or P < 0.01). The levels of Vimentin and TGF-β1 in lung tissue were significantly reduced, while p-AMPK level was significantly up-regulated (P < 0.05). Conclusion Exosomes derived from MSCs may alleviate RILI by inhibiting inflammatory responses and regulating epithelial-mesenchymal transition mediated by AMPK/TGF-β1 signaling pathway.
3.Mechanism of Traditional Chinese Medicine Regulating JAK/STAT Signaling Pathway to Intervene in Lung Cancer: A Review
Jiarui CAO ; Bo FENG ; Chunzheng MA ; Weixia CHEN ; Jiangfan YU ; Shasha CAO ; Zhenyu ZHANG ; Wenhui OUYANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):265-276
Lung cancer is the malignant tumor with the highest incidence and mortality rates globally. Current treatment methods for lung cancer primarily include surgery, chemotherapy, targeted therapy, and immunotherapy. However, the main limitations of these treatments are their side effects, the drug resistance, and the economic burden they impose. As a critical cancer pathway, the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway regulates tumor occurrence and development through multiple mechanisms by influencing various downstream targets. Consequently, the JAK/STAT signaling pathway offers a promising avenue for lung cancer treatment research. Numerous studies have demonstrated that the JAK/STAT signaling pathway plays a key role in the proliferation and growth of lung cancer cells, angiogenesis, epithelial-mesenchymal transition (EMT), metabolic alterations, remodeling of the immune microenvironment, and the development of treatment resistance. Traditional Chinese medicine (TCM) has garnered increasing attention due to its minimal side effects, low economic burden, and its potential to enhance efficacy and reduce toxicity when used in conjunction with Western medicine. In addition to traditional Chinese medicine compounds, a growing number of Chinese medicine monomers have come into the spotlight because of their more targeted effects. Numerous studies investigating the regulation of the JAK/STAT signaling pathway by TCM in the treatment of lung cancer have demonstrated that TCM can inhibit the proliferation and invasion of lung cancer cells, tumor angiogenesis, and EMT, improve the inflammatory and immunosuppressive microenvironments, and enhance treatment sensitivity by intervening in the JAK/STAT signaling pathway, thereby impeding the progression of lung cancer. In recent years, the research on the regulation of this pathway by TCM in the treatment of lung cancer has been updated rapidly. However, the summary of these studies has not been updated in time. This review summarizes and reflects on the recent research findings regarding the regulation of the JAK/STAT signaling pathway by TCM to intervene in lung cancer from three aspects, introducing the JAK/STAT pathway, elaborating the mechanism of this pathway in lung cancer, and exploring the intervention of TCM in the treatment of lung cancer through this pathway, to provide more reference for the treatment of lung cancer in the future.
4.Safety and effectiveness analysis of laparoscopic radical resection of colorectal cancer without indwelling gastric tube
Yu-Fei HU ; Heng LI ; Jun-Feng OUYANG ; Yan FU
Journal of Regional Anatomy and Operative Surgery 2024;33(12):1070-1073
Objective To evaluate the safety and effectiveness of laparoscopic radical resection of colorectal cancer without indwelling gastric tube during the perioperative period for postoperative rehabilitation of patients.Methods A total of 90 patients who underwent laparoscopic radical resection for colorectal cancer in our hospital from January 2021 to January 2023 were selected as the study objects,they were randomly divided into the observation group and the control group,with 45 cases in each group.The time of first postoperative exhaust and defecation,postoperative hospital stay,and the occurrence of complications such as throat pain,nausea and vomiting,pulmonary infection,intestinal obstruction,and anastomotic fistula of patients were compared between the two groups.Results The time of first postoperative exhaust and defecation and postoperative hospital stay of patients in the observation group were shorter than those in the control group,with statistically significant differences(P<0.05).The incidence of throat pain and nausea and vomiting of patients in the observation group was lower than that in the control group,with statistically significant difference(P<0.05);However,there was no statistically significant difference in postoperative complications such as pulmonary infection,intestinal obstruction,and anastomotic fistula of patients between the two groups(P>0.05).Conclusion Laparoscopic radical resection of colorectal cancer without indwelling gastric tube during the perioperative period will not increase the incidence of postoperative complications such as pulmonary infection,intestinal obstruction,and anastomotic fistula,instead,it can reduce the incidence of postoperative throat pain and nausea and vomiting,shorten hospital stay,and accelerate postoperative recovery of patients.
5.Successful Pulsed-field Ablation for Atrial Fibrillation Guided by Intracardiac Echocardiography and 3-Dimentional Mapping System:a Case Report
Guodong NIU ; Wenbin OUYANG ; Zhiling LUO ; Yu QIAO ; Mingpeng FU ; Yulong GUO ; Jinrui GUO ; Ke YANG ; Xiangbin PAN
Chinese Circulation Journal 2024;39(11):1133-1136
The present case report described a patient with paroxysmal atrial fibrillation who received pulsed-filed ablation guided by intracardiac echocardiography and 3-dimentional mapping system.All four pulmonary veins were isolated in the procedure,good clinical results and acute safety profile were achieved.The present case reveals the safety and feasibility of the technique for the treatment of paroxysmal atrial fibrillation.
6.Nutritional status and related factors of elderly hospitalized patients with hypertension and frailty
Shujuan ZHENG ; Yu LI ; Juyan OUYANG ; Tian TIAN ; Miyesai·Ainiwaer ; Yanan WANG ; Hong WANG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(8):886-891
Objective To survey the nutritional status of elderly hospitalized patients with hyper-tension and frailty,and explore the correlation between nutrition and frailty.Methods A total of 801 elderly hypertensive patients admitted to our hospital from October 2022 to December 2023 were cosecutively enrolled,and according to Fried frailty phenotype criteria,they were divided into frailty group(score:≥3,276 cases),pre-frailty group(score:1-3,362 cases)and non-frailty group(score:0,163 cases).At the same time,MNA was used to evaluate the nutritional risk of the patients,and the scores of ADL and the score of ACCI were recorded.Logistic regression mod-el was used to analyze the influencing factors of frailty in elderly hypertensive patients,Spearman correlation analysis was employed to analyze the relationship between frailty and nutrition-related indicators,and ROC curve was plotted to analyze the predictive performance of these nutritional indicators in the occurrence of frailty.Results There were statistically differences among the three groups in terms of age,mean SBP,complication of other chronic diseases,polypharmacy,mean handgrip strength,calf circumference,MNA score and classification,ADL and ACCI scores,and levels of hemoglobin,HDL-C,LDL-C,TC,albumin,25-hydroxyvitamin D[25(OH)D],bone alkaline phosphatase,VitB12 and Hcy(P<0.05,P<0.01).Multivariate logistic regression analysis revealed that polypharmacy(OR=2.361,95%CI:1.332-4.183),MNA score ≥24(OR=0.298,95%CI:0.110-0.808),ADL score(OR=1.094,95%CI:1.028-1.166),albumin(OR=0.963,95%CI:0.934-0.994),and 25(OH)D(OR=0.989,95%CI:0.980-0.998)were independent risk factors for frailty in elderly hypertensive patients(P<0.05,P<0.01).Frailty was positively cor-related with age,polypharmacy,other chronic diseases,and ACCI score(P<0.01),and negatively with MNA score,MNA classification,hemoglobin,albumin,25(OH)D,HDL-C,LDL-C,Hcy,average grip strength,and calf circumference(P<0.01).ROC curve analysis showed that the AUC values of 25(OH)D,albumin,and MNA score were 0.607,0.588,and 0.700,separately.Con-clusion Elderly patients with hypertension and frailty have poor nutritional status,and identify-ing early frailty relies on nutritional risk assessment.Frail patients have a high risk of malnutri-tion and are influenced by various factors.Among them,albumin,MNA score,and 25(OH)D can effectively predict the occurrence of frailty in elderly hospitalized hypertensive patients,with MNA score having stronger predictive performance.
7.Intraoperative frozen section pathology of vaginal margin in radical hysterectomy on the prognosis and quality of life for patients with IB2–IIA2cervical cancer: study protocol for a multicenter randomized controlled trial
Yu LIU ; Weijuan XIN ; Ping WANG ; Mei JI ; Xiaoqing GUO ; Yunyan OUYANG ; Dong ZHAO ; Keqin HUA
Journal of Gynecologic Oncology 2024;35(3):e81-
Background:
Several risk factors have been identified that compromise the treatment outcome in patients with early-to-mid-stage cervical cancer (CC) who are primarily treated with radical surgery. However, there is no report on the impact of intraoperative frozen pathology examination of vaginal margins on the prognosis of patients with CC. This study aimed to conduct a randomized controlled trial (RCT) to determine whether selective vaginal resection can reduce the incidence of operative complications and the risk of postoperative radiotherapy. The impact of the length of the vagina removed in radical hysterectomy (RH) on prognosis and quality of life (QoL) for IB2–IIA2 CC patients will be investigated.
Methods
A multicenter, non-inferiority, RCT at 7 institutions in China is designed to investigate the effect of intraoperative frozen pathology exam of vaginal margin in RH on the survival outcomes for patients with IB2–IIA2 CC. Eligible patients aged 18–70 years will be randomly assigned online by one-to-one random allocation to receive intraoperative frozen pathology exam of vaginal margin or not. If frozen pathology indicates positive margin, continue resection of 1 centimeter of vaginal tissue until negative margin is achieved. The primary end point is 2-year disease-free survival (DFS). Adverse events (AEs) caused by further vagina resection, 5-year DFS, 2-year overall survival (OS), 5-year OS and AEs caused by radiotherapy and QoL are secondary end points. A total of 310 patients will be enrolled from 7 tertiary hospitals in China within 3-year period and followed up for 5 years.Trial Registration: Chinese Clinical Trial Registry Identifier: ChiCTR2000035668
8.Impact of serum vitamin A levels on all-cause mortality risk in diabetes patients:a prospective study
Zhuo LI ; Baoyuan JIANG ; Yu OUYANG ; Yaxin XIANG ; Zhen YANG ; Lianying GUO
Journal of Shenyang Medical College 2024;26(6):575-581
Objective:To investigate the impact of serum vitamin A levels on all-cause mortality risk in diabetes patients.Methods:Diabetes patients aged 20 years and above who participated in the US National Health and Nutrition Examination Survey during 2003-2004 and 2005-2006 were enrolled as the study population,with death data up to 2019 as the endpoint.Cox proportional hazards regression models were employed to calculate the hazard ratios of all-cause mortality in diabetes patients with different serum vitamin A levels,both unadjusted and adjusted for confounders.Restricted cubic spline methods were used to analyze the dose-response relationship between serum vitamin A levels and all-cause mortality risk in diabetes patients.Results:A total of 484 diabetes patients were included,with a median follow-up period of 13.7 years,during which 211 deaths occurred.Cox proportional hazards analysis showed that compared to the lowest quartile of serum vitamin A,higher quartiles of serum vitamin A were not associated with all-cause mortality risk in diabetes patients without adjusting for confounders.However,after adjusting for confounders,higher quartiles of serum vitamin A significantly reduced the all-cause mortality risk.The dose-response analysis indicated a significantly increased risk of all-cause mortality in diabetes patients with lower serum vitamin A levels.As the vitamin A levels increased,the mortality risk gradually decreased.A significant reduction in all-cause mortality risk was observed when serum vitamin A levels were between 2.17 and 2.50 μmol/L.Beyond this range,there was a tendency for increased all-cause mortality risk with further increases in vitamin A levels.Conclusion:Lower serum vitamin A levels increase the all-cause mortality risk in diabetes patients,while moderate serum vitamin A levels help reduce the all-cause mortality risk in diabetes patients.
9.Analysis of Research Hotspots and Theme Evolution in Traditional Chinese Medicine Based on BERTopic and CiteSpace
Anli WANG ; Wenting ZHANG ; Tingxiao WEN ; Wei OUYANG ; Shanshan YU ; Feilong YANG
Journal of Medical Informatics 2024;45(7):55-61
Purpose/Significance To sort out the literature in the field of traditional Chinese medicine(TCM)in China in the past decade,and to expound the hot topics and development trends in this field.Method/Process Literature in the field of TCM in Wanfang Data from 2013 to 2022 is searched.The abstracts are taken as the research objects,the topics are extracted by BERTopic model,and CiteSpace is used for visualization and analysis.Result/Conclusion 12 683 papers in Chinese are included,and the 5 topics are found:clinical trials and efficacy,research on pathogenesis and pharmacology,the development of the field of TCM,treatment of chronic dis-ease,drug analysis and literature research.The paper makes an in-depth analysis of the topic of the development of traditional Chinese medicine,which continues to grow in popularity,and puts forward that the deep integration of artificial intelligence(AI)and TCM is the future trend.
10.Construction of the prediction model of breast cancer bone metastasis based on machine learning
Fei OUYANG ; Yang WANG ; Yu CHEN ; Guoqing PEI ; Ling WANG ; Yang ZHANG ; Lei SHI
China Oncology 2024;34(10):903-914
Background and purpose:Breast cancer is a major global public health problem.Bone is the most common site of distant metastasis of breast cancer,accounting for about 70%of all metastatic cases.Bone metastasis of breast cancer can cause a series of complications,including severe pain,pathological fracture,hypercalcemia,spinal cord compression,etc.,which bring great inconvenience to patients'physical activities and affect their quality of life.Metastatic recurrence is the leading cause of death in breast cancer patients.Therefore,there is an urgent need to build a diagnostic model of bone metastasis in breast cancer to identify patients with a high risk of bone metastasis.The aim of this study was to develop a predictive model based on machine learning to predict the probability of breast cancer developing bone metastasis.Methods:Data of breast cancer patients diagnosed between 2010 and 2015 were extracted from The Surveillance,Epidemiology,and End Results(SEER)database.The variables were screened by least absolute shrinkage and selection operator(LASSO)regression,univariate and multivariate logistic regression analysis,and statistically significant risk factors were included to build a prediction model.In this study,nine machine learning algorithms,including decision tree,elastic network,K-nearest neighbor,lightweight gradient elevator,logistic regression,neural network,random forest,support vector machine and limit gradient lifting,were used to adjust the model hyperparameters through random search and 5x cross-validation to build a breast cancer bone metastasis prediction model.The area under the receiver operating characteristic(ROC)curve,calibration curve and decision curve were used to evaluate the model,the optimal model was obtained,and the importance of variables was analyzed based on the optimal model.Finally,a network calculator for predicting the risk of bone metastasis of breast cancer was established using the optimal model.Results:The study included 10 106 patients with breast cancer,7 073 patients in the training set,and 3 033 patients in the validation set.We found that 4 494(63.5%)patients in the training set and 1 927(63.5%)patients in the validation set developed bone metastases,respectively.Race,pathologic grade,estrogen receptor(ER)status,progesterone receptor(PR)status,human epidermal growth factor receptor 2(HER2)status,N stage,lung metastasis,radiotherapy,chemotherapy and surgery were independent predictors of bone metastasis.The training set and verification set were used to verify the model,and the limit gradient lifting algorithm was superior to other machine learning algorithms by integrating the evaluation indexes such as the area under the ROC curve,calibration curve and decision curve.Finally,we used limit gradient algorithm to build network calculator for prediction of breast cancer bone metastases(https://bcbm.shinyapps.io/DynNomapp/).Conclusion:This study developed a predictive model based on machine learning to predict the probability of bone metastases in breast cancer patients,hoping to help clinicians make more rational treatment decisions.

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