1.Effect of Liuwei Dihuangwan on EMT and Expression of CSC Properties in 4T1 Cells by Regulating Myeloid-derived Suppressor Cells
Lixiang ZHENG ; Ling HUANG ; Huiwen GUO ; Biyao GONG ; Xiaoying REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):1-10
ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum (LDP) on epithelial-mesenchymal transition (EMT) and the expression of cancer stem cell (CSC) properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells (MDSCs). MethodsSPF-grade female SD rats were randomly divided into three groups, which were given 0.39, 1.94, 3.89 g·kg-1·d-1 suspension of Liuwei Dihuangwan for 7 days, respectively, to prepare low-, medium-, and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 (CCK-8) assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry (FC). The migration, invasion, and proliferation of 4T1 cells were detected by cell scratch assay, Transwell invasion assay, and plate colony-forming assay, respectively. Western blot (WB) was used to detect the protein expression of transforming growth factor-β (TGF-β), nuclear factor-κB (NF-κB), C-X-C motif chemokine ligand 2 (CXCL2), E-cadherin, and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence (IF). ResultsCompared with the normal group, LDP showed no significant inhibitory effect on the cell viability of 4T1 cells, but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs, as well as the expression of TGF-β (P<0.05, P<0.01). The migration, invasion, and proliferation of 4T1 cells were increased after co-culture with MDSCs (P<0.05, P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin and the proportion of CSC (CD44+CD24-) were elevated (P<0.05, P<0.01), while the expression of E-cadherin was decreased (P<0.05). After the intervention of MDSCs with LDP, followed by co-culture with 4T1 cells, the migration, invasion, and proliferation of 4T1 cells were obviously reduced (P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin were decreased (P<0.05, P<0.01), and the expression of E-cadherin was increased (P<0.05, P<0.01). There was no statistical difference in the proportion of CSC (CD44+CD24-) in 4T1 cells. However, the proportion of CSC (CD44+CD24-) was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention (P<0.05, P<0.01). ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β, NF-κB, and CXCL2, reverse EMT, and reduce the characteristic expression of CSC to inhibit the migration, invasion, and proliferation of 4T1 cells.
2.Effect of Liuwei Dihuangwan on EMT and Expression of CSC Properties in 4T1 Cells by Regulating Myeloid-derived Suppressor Cells
Lixiang ZHENG ; Ling HUANG ; Huiwen GUO ; Biyao GONG ; Xiaoying REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):1-10
ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum (LDP) on epithelial-mesenchymal transition (EMT) and the expression of cancer stem cell (CSC) properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells (MDSCs). MethodsSPF-grade female SD rats were randomly divided into three groups, which were given 0.39, 1.94, 3.89 g·kg-1·d-1 suspension of Liuwei Dihuangwan for 7 days, respectively, to prepare low-, medium-, and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 (CCK-8) assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry (FC). The migration, invasion, and proliferation of 4T1 cells were detected by cell scratch assay, Transwell invasion assay, and plate colony-forming assay, respectively. Western blot (WB) was used to detect the protein expression of transforming growth factor-β (TGF-β), nuclear factor-κB (NF-κB), C-X-C motif chemokine ligand 2 (CXCL2), E-cadherin, and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence (IF). ResultsCompared with the normal group, LDP showed no significant inhibitory effect on the cell viability of 4T1 cells, but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs, as well as the expression of TGF-β (P<0.05, P<0.01). The migration, invasion, and proliferation of 4T1 cells were increased after co-culture with MDSCs (P<0.05, P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin and the proportion of CSC (CD44+CD24-) were elevated (P<0.05, P<0.01), while the expression of E-cadherin was decreased (P<0.05). After the intervention of MDSCs with LDP, followed by co-culture with 4T1 cells, the migration, invasion, and proliferation of 4T1 cells were obviously reduced (P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin were decreased (P<0.05, P<0.01), and the expression of E-cadherin was increased (P<0.05, P<0.01). There was no statistical difference in the proportion of CSC (CD44+CD24-) in 4T1 cells. However, the proportion of CSC (CD44+CD24-) was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention (P<0.05, P<0.01). ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β, NF-κB, and CXCL2, reverse EMT, and reduce the characteristic expression of CSC to inhibit the migration, invasion, and proliferation of 4T1 cells.
3.Chain mediating roles of perceived social support and coping modes between perceived risk of recurrence and depression in elderly patients with first-episode ischemic stroke
Yang SHI ; Zhie GU ; Hailong YU ; Tong ZHU ; Huiwen HUANG ; Lin WANG
Chinese Journal of Modern Nursing 2025;31(4):527-533
Objective:To explore the chain mediating role of perceived social support and coping modes between perceived risk of recurrence and depression in elderly patients with first-episode ischemic stroke.Methods:Convenience sampling was used to select 310 elderly patients with first-episode ischemic stroke in the Department of Neurology of Jiangsu Wutaishan Hospital and Northern Jiangsu People 's Hospital from December 2022 to February 2024 for the study. General Information Questionnaire, Recurrence Risk Perception Scale for Patients with Stroke, Multidimensional Scale of Perceived Social Support, Medical Coping Modes Questionnaire, and Patient Health Questionnaire were used to investigate the patients and to construct and validate the chain mediating model. Results:A total of 310 questionnaires were distributed and 306 valid questionnaires were recovered, with a valid recovery rate of 98.7%. The total scores of perceived risk of recurrence, perceived social support, medical coping modes, and depression in elderly patients with first-episode ischemic stroke were 35.00 (29.00, 39.00), 59.00 (51.00, 67.00), 43.00 (39.00, 47.00), and 3.50 (2.00, 7.00), respectively. Perceived risk of recurrence was positively correlated with perceived social support and confrontation coping mode ( r=0.418, 0.463; P<0.001), and negatively correlated with depression ( r=-0.450, P<0.001). Perceived social support was positively correlated with confrontation coping mode ( r=0.416, P<0.001), and negatively correlated with depression ( r=-0.408, P<0.001). Confrontation coping mode was negatively correlated with depression ( r=-0.472, P<0.001). The mediating test showed that perceived risk of recurrence negatively predicted depression, with a direct effect of -0.245, accounting for 48.70% of the total effect. The mediating effect of perceived social support on perceived risk of recurrence and depression was significant, with an indirect effect of -0.146, accounting for 29.03% of the total effect. The mediating effect of confrontation coping mode on perceived risk of recurrence and depression was significant, with an indirect effect of -0.085, accounting for 16.90% of the total effect. In addition, perceived social support and confrontation coping modes had a significant chain mediating effect in perceived risk of recurrence and depression, with an indirect effect of -0.027, accounting for 5.37% of the total effect. Conclusions:Perceived social support and confrontation coping mode have chain mediating effects between perceived risk of recurrence and depression. Healthcare professionals can reduce the incidence of depression by increasing the patient 's perceived risk of recurrence, giving a great deal of social support, and enhancing positive modes of coping with the disease.
4.Trajectory and influencing factors of oral frailty in first-onset elderly patients with ischemic stroke
Yang SHI ; Hailong YU ; Zhie GU ; Huiwen HUANG ; Li JI ; Lin WANG ; Can CHEN
Chinese Journal of Modern Nursing 2025;31(16):2195-2202
Objective:To explore the trajectory and influencing factors of oral frailty in first-onset elderly patients with ischemic stroke.Methods:A total of 240 inpatients from the neurology departments of Wutaishan Hospital and Northern Jiangsu People's Hospital between October 2022 and February 2024 were selected by convenience sampling. Oral frailty scores were assessed dynamically at four time points: 1 day before discharge (T 1) , 1 month after discharge (T 2) , 3 months after discharge (T 3) , and 6 months after discharge (T 4) . Latent growth modeling was used to identify potential trajectory categories and analyze their influencing factors. Results:A total of 240 questionnaires were distributed, with 220 valid responses, yielding a response rate of 91.67%. A total of three trajectory types of oral frailty were identified: non-oral frailty group ( n=92) , low-decline group ( n=66) , and high-decline group ( n=62) . Age, marital status, smoking history, dysphagia, National Institute of Health Stroke Scale score, oral health, nutritional status, oral health-related self-efficacy, monthly per capita household income, polypharmacy, denture use, and cognitive function were influencing factors of oral frailty in first-onset elderly patients with ischemic stroke ( P<0.05) . Conclusions:From 1 day before discharge to 6 months after discharge, there are 3 distinct oral frailty trajectories in first-onset elderly patients with ischemic stroke. Medical staff should implement individualized interventions based on the identified influencing factors.
5.Chain mediating roles of perceived social support and coping modes between perceived risk of recurrence and depression in elderly patients with first-episode ischemic stroke
Yang SHI ; Zhie GU ; Hailong YU ; Tong ZHU ; Huiwen HUANG ; Lin WANG
Chinese Journal of Modern Nursing 2025;31(4):527-533
Objective:To explore the chain mediating role of perceived social support and coping modes between perceived risk of recurrence and depression in elderly patients with first-episode ischemic stroke.Methods:Convenience sampling was used to select 310 elderly patients with first-episode ischemic stroke in the Department of Neurology of Jiangsu Wutaishan Hospital and Northern Jiangsu People 's Hospital from December 2022 to February 2024 for the study. General Information Questionnaire, Recurrence Risk Perception Scale for Patients with Stroke, Multidimensional Scale of Perceived Social Support, Medical Coping Modes Questionnaire, and Patient Health Questionnaire were used to investigate the patients and to construct and validate the chain mediating model. Results:A total of 310 questionnaires were distributed and 306 valid questionnaires were recovered, with a valid recovery rate of 98.7%. The total scores of perceived risk of recurrence, perceived social support, medical coping modes, and depression in elderly patients with first-episode ischemic stroke were 35.00 (29.00, 39.00), 59.00 (51.00, 67.00), 43.00 (39.00, 47.00), and 3.50 (2.00, 7.00), respectively. Perceived risk of recurrence was positively correlated with perceived social support and confrontation coping mode ( r=0.418, 0.463; P<0.001), and negatively correlated with depression ( r=-0.450, P<0.001). Perceived social support was positively correlated with confrontation coping mode ( r=0.416, P<0.001), and negatively correlated with depression ( r=-0.408, P<0.001). Confrontation coping mode was negatively correlated with depression ( r=-0.472, P<0.001). The mediating test showed that perceived risk of recurrence negatively predicted depression, with a direct effect of -0.245, accounting for 48.70% of the total effect. The mediating effect of perceived social support on perceived risk of recurrence and depression was significant, with an indirect effect of -0.146, accounting for 29.03% of the total effect. The mediating effect of confrontation coping mode on perceived risk of recurrence and depression was significant, with an indirect effect of -0.085, accounting for 16.90% of the total effect. In addition, perceived social support and confrontation coping modes had a significant chain mediating effect in perceived risk of recurrence and depression, with an indirect effect of -0.027, accounting for 5.37% of the total effect. Conclusions:Perceived social support and confrontation coping mode have chain mediating effects between perceived risk of recurrence and depression. Healthcare professionals can reduce the incidence of depression by increasing the patient 's perceived risk of recurrence, giving a great deal of social support, and enhancing positive modes of coping with the disease.
6.Trajectory and influencing factors of oral frailty in first-onset elderly patients with ischemic stroke
Yang SHI ; Hailong YU ; Zhie GU ; Huiwen HUANG ; Li JI ; Lin WANG ; Can CHEN
Chinese Journal of Modern Nursing 2025;31(16):2195-2202
Objective:To explore the trajectory and influencing factors of oral frailty in first-onset elderly patients with ischemic stroke.Methods:A total of 240 inpatients from the neurology departments of Wutaishan Hospital and Northern Jiangsu People's Hospital between October 2022 and February 2024 were selected by convenience sampling. Oral frailty scores were assessed dynamically at four time points: 1 day before discharge (T 1) , 1 month after discharge (T 2) , 3 months after discharge (T 3) , and 6 months after discharge (T 4) . Latent growth modeling was used to identify potential trajectory categories and analyze their influencing factors. Results:A total of 240 questionnaires were distributed, with 220 valid responses, yielding a response rate of 91.67%. A total of three trajectory types of oral frailty were identified: non-oral frailty group ( n=92) , low-decline group ( n=66) , and high-decline group ( n=62) . Age, marital status, smoking history, dysphagia, National Institute of Health Stroke Scale score, oral health, nutritional status, oral health-related self-efficacy, monthly per capita household income, polypharmacy, denture use, and cognitive function were influencing factors of oral frailty in first-onset elderly patients with ischemic stroke ( P<0.05) . Conclusions:From 1 day before discharge to 6 months after discharge, there are 3 distinct oral frailty trajectories in first-onset elderly patients with ischemic stroke. Medical staff should implement individualized interventions based on the identified influencing factors.
7.Research idea on the importance of traditional Chinese medicine tongue inspection based on inference correction
Jiale DOU ; Huiwen HUANG ; Tong WU ; Chunhua JIA
Journal of Beijing University of Traditional Chinese Medicine 2024;47(9):1223-1228
Previous research on traditional Chinese medicine tongue inspection has mainly focused on its development,clinical application,and objectification,ignoring the fundamental question of "why tongue inspection is important in traditional Chinese medicine diagnosis" and neglecting the dominant role of traditional Chinese medicine brain thinking activities during diagnosis. Therefore,applying modern cognitive neuroscience technology to explore the impact of tongue inspection information on traditional Chinese medicine diagnostic thinking from an empirical research perspective may open up a new path for future tongue inspection research. This article proposes the combined application of eye tracking and event-related potential technology,drawing on the experimental paradigm of text reading inference correction in cognitive psychology,to collect eye movement and electroencephalogram data of participants induced by changes in tongue information during the reading process of traditional Chinese medicine medical records. This method can achieve synchronous observation of the focus areas and electroencephalogram signals of the subjects during the reading process,and provide cognitive scientific evidence for "why tongue inspection is important in traditional Chinese medicine diagnosis" through statistical data analysis. This research approach integrates the theories,techniques,and methods of traditional Chinese medicine and cognitive science,and is of great significance for developing cognitive thinking research in traditional Chinese medicine.
8.A multicenter prospective study on early identification of refractory Mycoplasma pneumoniae pneumonia in children
Dan XU ; Ailian ZHANG ; Jishan ZHENG ; Mingwei YE ; Fan LI ; Gencai QIAN ; Hongbo SHI ; Xiaohong JIN ; Lieping HUANG ; Jiangang MEI ; Guohua MEI ; Zhen XU ; Hong FU ; Jianjun LIN ; Hongzhou YE ; Yan ZHENG ; Lingling HUA ; Min YANG ; Jiangmin TONG ; Lingling CHEN ; Yuanyuan ZHANG ; Dehua YANG ; Yunlian ZHOU ; Huiwen LI ; Yinle LAN ; Yulan XU ; Jinyan FENG ; Xing CHEN ; Min GONG ; Zhimin CHEN ; Yingshuo WANG
Chinese Journal of Pediatrics 2024;62(4):317-322
Objective:To explore potential predictors of refractory Mycoplasma pneumoniae pneumonia (RMPP) in early stage. Methods:The prospective multicenter study was conducted in Zhejiang, China from May 1 st, 2019 to January 31 st, 2020. A total of 1 428 patients with fever >48 hours to <120 hours were studied. Their clinical data and oral pharyngeal swab samples were collected; Mycoplasma pneumoniae DNA in pharyngeal swab specimens was detected. Patients with positive Mycoplasma pneumoniae DNA results underwent a series of tests, including chest X-ray, complete blood count, C-reactive protein, lactate dehydrogenase (LDH), and procalcitonin. According to the occurrence of RMPP, the patients were divided into two groups, RMPP group and general Mycoplasma pneumoniae pneumonia (GMPP) group. Measurement data between the 2 groups were compared using Mann-Whitney U test. Logistic regression analyses were used to examine the associations between clinical data and RMPP. Receiver operating characteristic (ROC) curves were used to analyse the power of the markers for predicting RMPP. Results:A total of 1 428 patients finished the study, with 801 boys and 627 girls, aged 4.3 (2.7, 6.3) years. Mycoplasma pneumoniae DNA was positive in 534 cases (37.4%), of whom 446 cases (83.5%) were diagnosed with Mycoplasma pneumoniae pneumonia, including 251 boys and 195 girls, aged 5.2 (3.3, 6.9) years. Macrolides-resistant variation was positive in 410 cases (91.9%). Fifty-five cases were with RMPP, 391 cases with GMPP. The peak body temperature before the first visit and LDH levels in RMPP patients were higher than that in GMPP patients (39.6 (39.1, 40.0) vs. 39.2 (38.9, 39.7) ℃, 333 (279, 392) vs. 311 (259, 359) U/L, both P<0.05). Logistic regression showed the prediction probability π=exp (-29.7+0.667×Peak body temperature (℃)+0.004×LDH (U/L))/(1+exp (-29.7+0.667×Peak body temperature (℃)+0.004 × LDH (U/L))), the cut-off value to predict RMPP was 0.12, with a consensus of probability forecast of 0.89, sensitivity of 0.89, and specificity of 0.67; and the area under ROC curve was 0.682 (95% CI 0.593-0.771, P<0.01). Conclusion:In MPP patients with fever over 48 to <120 hours, a prediction probability π of RMPP can be calculated based on the peak body temperature and LDH level before the first visit, which can facilitate early identification of RMPP.
9.Clinical prediction model for complicated appendicitis in children under five years old
Tianming WANG ; Guoqin ZHANG ; Tingjun LI ; Jiahu HUANG ; Zhagen WANG ; Huiwen TANG ; Zhujun GU ; Jian LIU ; Xingyuan LIU
Chinese Pediatric Emergency Medicine 2023;30(4):286-290
Objective:To retrospectively analyze the independent risk factors of complicated appendicitis(CA)in children under five years old and establish a clinical prediction model, and to evaluate the clinical application of this model.Methods:A retrospective analysis was performed on children under five years old who underwent appendectomy at Children′s Hospital of Shanghai Jiao Tong University School of Medicine from January 2018 to December 2021.The children were divided into CA group and uncomplicated appendicitis group according to whether there was sign of perforation or gangrene in appendiceal tissue after operation.The differences in clinical features and preoperative laboratory test results between two groups were compared.The independent risk factors of CA were identified and a clinical prediction model was established.The clinical prediction model was verified by receiver operating characteristic curve.Results:A total of 140 children were enrolled in this study, including 84 cases in the CA group and 56 cases in uncomplicated appendicitis group.Univariate and binary Logistic regression analysis showed that the duration of symptoms>23.5 h( OR=6.650, 95% CI 2.469-17.912, P<0.05), abdominal muscle tension( OR=3.082, 95% CI 1.190-7.979, P<0.05) and C-reactive protein>41 mg/L ( OR=3.287, 95% CI 1.274-8.480, P<0.05) were independent risk factors for CA( P<0.05). The clinical prediction model of CA was constructed by the above mentioned three independent risk factors.The area under the receiver operating characteristic curve of the clinical prediction model was 0.881(95% CI 0.825-0.936), the sensitivity was 77.4%, the specificity was 87.5%, the positive predictive value was 91.3% and the negative predictive value was 70.0%. Conclusion:Acute appendicitis in children under five years old is more likely to progress to CA if the duration of symptoms>23.5 h, the level of C-reactive protein is increased, and the abdominal muscle tension is accompanied.The clinical prediction model of CA constructed by common clinical information in pediatric clinics has good prediction efficiency, which provides a simple and feasible reference method for clinicians to distinguish CA from uncomplicated appendicitis.
10.Analysis and suggestions on China′s Traditional Chinese Medicine health service policies from the perspective of policy life cycle
Chinese Journal of Hospital Administration 2023;39(8):553-557
Objective:To analyse China′s Traditional Chinese Medicine(TCM) health service policy from the perspective of policy life cycle, so as to provide reference for improving the policy system of TCM health service.Methods:The official websites of national government agencies were searched for policy documents on TCM health service from 2012 to 2022, using the keywords of " Chinese medicine" " Traditional Chinese Medicine" and " health service", and policy text entries were extracted and coded. A two-dimensional analytical framework of policy tools(supply, demand, and environment) and policy life cycle(agenda setting, policy formulation, policy implementation, and policy evaluation) was constructed to quantitatively analyze the policy texts.Results:A total of 58 policy documents were included, containing 2 898 text entries. The distribution of policy instruments was uneven, with a high percentage of entries using supply-based instruments(40.57%, 1 176 entries), followed by environmental instruments(38.86%, 1 126 entries) and demand-based instruments(20.57%, 596 entries). From the perspective of policy cycle, the policy tools were mainly concentrated in the stages of policy formulation(53.77%, 1 558 entries) and policy implementation(36.86%, 1 068 entries), while the policy tools in the stages of agenda setting(4.97%, 144 entries) and policy evaluation(4.40%, 128 entries) needed to be strengthened.Conclusions:In recent years, the distribution of policy tools in China′s TCM health service policy was uneven, and the articulation of the stages in the policy life cycle needed to be strengthened.

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