1.Isolation,culture and differentiation of human urine-derived stem cells into smooth muscle cells
Jiahui CHEN ; Xiaoqi DAI ; Yangang XU ; Yuanchao LI ; Mei HUANG ; Yifei ZHAN ; Yuxuan DU ; Liuqiang LI ; Yaochuan GUO ; Jun BIAN ; Dehui LAI
Chinese Journal of Tissue Engineering Research 2025;29(19):4076-4082
		                        		
		                        			
		                        			BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.
		                        		
		                        		
		                        		
		                        	
2.Daurisoline Inhibits Progression of Triple-Negative Breast Cancer by Regulating the γγ-Secretase/Notch Axis
Xiangyi ZHAN ; Xiaoyong CHEN ; Mei FENG ; Kuo YAO ; Kefan YANG ; Hui JIA
Biomolecules & Therapeutics 2025;33(2):331-343
		                        		
		                        			
		                        			 Triple-negative breast cancer (TNBC) is a subtype of breast cancer that is challenging to treat and lacks targeted therapeutic drugs in the clinic. Natural active ingredients provide promising opportunities for discovering and developing targeted therapies for TNBC. This study investigated the effects of daurisoline on TNBC and elucidated its potential mechanisms. Using network pharmacology, a correlation was identified between daurisoline, derived from Menispermum dauricum, and breast cancer, particularly involving the Notch signaling pathway. The effects of daurisoline on the proliferation, migration, and apoptosis of MDA-MB-231 and MDA-MB-468 cells were evaluated in vitro. Additionally, the impact of daurisoline on the growth of MDA-MB-231 xenograft tumors in nude mice was assessed through in vivo experiments. Expression levels of Notch signaling pathway-related proteins, including Notch-1, NICD, PSEN-1, Bax, and Bcl-2, were examined using molecular docking and Western blotting to explore the underlying mechanisms of daurisoline’s anti-breast cancer effects. It was revealed that daurisoline could effectively inhibit the proliferation and migration of MDA-MB-231 and MDA-MB-468 cells and promote apoptosis. Furthermore, it significantly reduced the growth of subcutaneous tumors in nude mice. Notably, daurisoline could reduce the hydrolytic activity of γ-secretase by binding to the catalytic core PSEN-1, thereby inhibiting activation of the γ-secretase/Notch axis and contributing to its anti-TNBC effects.This study supported the development of naturally targeted drugs for TNBC and provided insights into the research on dibenzylisoquinoline alkaloids, such as daurisoline. 
		                        		
		                        		
		                        		
		                        	
3.Daurisoline Inhibits Progression of Triple-Negative Breast Cancer by Regulating the γγ-Secretase/Notch Axis
Xiangyi ZHAN ; Xiaoyong CHEN ; Mei FENG ; Kuo YAO ; Kefan YANG ; Hui JIA
Biomolecules & Therapeutics 2025;33(2):331-343
		                        		
		                        			
		                        			 Triple-negative breast cancer (TNBC) is a subtype of breast cancer that is challenging to treat and lacks targeted therapeutic drugs in the clinic. Natural active ingredients provide promising opportunities for discovering and developing targeted therapies for TNBC. This study investigated the effects of daurisoline on TNBC and elucidated its potential mechanisms. Using network pharmacology, a correlation was identified between daurisoline, derived from Menispermum dauricum, and breast cancer, particularly involving the Notch signaling pathway. The effects of daurisoline on the proliferation, migration, and apoptosis of MDA-MB-231 and MDA-MB-468 cells were evaluated in vitro. Additionally, the impact of daurisoline on the growth of MDA-MB-231 xenograft tumors in nude mice was assessed through in vivo experiments. Expression levels of Notch signaling pathway-related proteins, including Notch-1, NICD, PSEN-1, Bax, and Bcl-2, were examined using molecular docking and Western blotting to explore the underlying mechanisms of daurisoline’s anti-breast cancer effects. It was revealed that daurisoline could effectively inhibit the proliferation and migration of MDA-MB-231 and MDA-MB-468 cells and promote apoptosis. Furthermore, it significantly reduced the growth of subcutaneous tumors in nude mice. Notably, daurisoline could reduce the hydrolytic activity of γ-secretase by binding to the catalytic core PSEN-1, thereby inhibiting activation of the γ-secretase/Notch axis and contributing to its anti-TNBC effects.This study supported the development of naturally targeted drugs for TNBC and provided insights into the research on dibenzylisoquinoline alkaloids, such as daurisoline. 
		                        		
		                        		
		                        		
		                        	
4.Daurisoline Inhibits Progression of Triple-Negative Breast Cancer by Regulating the γγ-Secretase/Notch Axis
Xiangyi ZHAN ; Xiaoyong CHEN ; Mei FENG ; Kuo YAO ; Kefan YANG ; Hui JIA
Biomolecules & Therapeutics 2025;33(2):331-343
		                        		
		                        			
		                        			 Triple-negative breast cancer (TNBC) is a subtype of breast cancer that is challenging to treat and lacks targeted therapeutic drugs in the clinic. Natural active ingredients provide promising opportunities for discovering and developing targeted therapies for TNBC. This study investigated the effects of daurisoline on TNBC and elucidated its potential mechanisms. Using network pharmacology, a correlation was identified between daurisoline, derived from Menispermum dauricum, and breast cancer, particularly involving the Notch signaling pathway. The effects of daurisoline on the proliferation, migration, and apoptosis of MDA-MB-231 and MDA-MB-468 cells were evaluated in vitro. Additionally, the impact of daurisoline on the growth of MDA-MB-231 xenograft tumors in nude mice was assessed through in vivo experiments. Expression levels of Notch signaling pathway-related proteins, including Notch-1, NICD, PSEN-1, Bax, and Bcl-2, were examined using molecular docking and Western blotting to explore the underlying mechanisms of daurisoline’s anti-breast cancer effects. It was revealed that daurisoline could effectively inhibit the proliferation and migration of MDA-MB-231 and MDA-MB-468 cells and promote apoptosis. Furthermore, it significantly reduced the growth of subcutaneous tumors in nude mice. Notably, daurisoline could reduce the hydrolytic activity of γ-secretase by binding to the catalytic core PSEN-1, thereby inhibiting activation of the γ-secretase/Notch axis and contributing to its anti-TNBC effects.This study supported the development of naturally targeted drugs for TNBC and provided insights into the research on dibenzylisoquinoline alkaloids, such as daurisoline. 
		                        		
		                        		
		                        		
		                        	
5.Visualization Analysis of Artificial Intelligence Literature in Forensic Research
Yi-Ming DONG ; Chun-Mei ZHAO ; Nian-Nian CHEN ; Li LUO ; Zhan-Peng LI ; Li-Kai WANG ; Xiao-Qian LI ; Ting-Gan REN ; Cai-Rong GAO ; Xiang-Jie GUO
Journal of Forensic Medicine 2024;40(1):1-14
		                        		
		                        			
		                        			Objective To analyze the literature on artificial intelligence in forensic research from 2012 to 2022 in the Web of Science Core Collection Database,to explore research hotspots and developmen-tal trends.Methods A total of 736 articles on artificial intelligence in forensic medicine in the Web of Science Core Collection Database from 2012 to 2022 were visualized and analyzed through the litera-ture measuring tool CiteSpace.The authors,institution,country(region),title,journal,keywords,cited references and other information of relevant literatures were analyzed.Results A total of 736 articles published in 220 journals by 355 authors from 289 institutions in 69 countries(regions)were identi-fied,with the number of articles published showing an increasing trend year by year.Among them,the United States had the highest number of publications and China ranked the second.Academy of Forensic Science had the highest number of publications among the institutions.Forensic Science Inter-national,Journal of Forensic Sciences,International Journal of Legal Medicine ranked high in publica-tion and citation frequency.Through the analysis of keywords,it was found that the research hotspots of artificial intelligence in the forensic field mainly focused on the use of artificial intelligence technol-ogy for sex and age estimation,cause of death analysis,postmortem interval estimation,individual identification and so on.Conclusion It is necessary to pay attention to international and institutional cooperation and to strengthen the cross-disciplinary research.Exploring the combination of advanced ar-tificial intelligence technologies with forensic research will be a hotspot and direction for future re-search.
		                        		
		                        		
		                        		
		                        	
6.Clinical Diseases Responding Specifically to Traditional Chinese Medicine:Chronic Prostatitis
Zhan QIN ; Mei MO ; Chaoba HE ; Liguo GENG ; Bin WANG ; Jisheng WANG ; Kecheng LI ; Nianwen HUANG ; Zhiming HONG ; Shaoying YUAN ; Xiaoxiao ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):152-158
		                        		
		                        			
		                        			Chronic prostatitis is a common and frequently occurring disease in male clinics. Due to the diverse symptoms, complex etiology, unclear mechanisms, and recurrent episodes of chronic prostatitis, Western medicine often faces the challenge of limited treatment options and unsatisfactory outcomes. In order to fully leverage the unique advantages of traditional Chinese medicine (TCM) in the treatment of chronic prostatitis, the China Association of Chinese Medicine organized a discussion among experts from the fields of TCM urology, Western urology, and related pharmacy. The experts focused on the advancements in the understanding of chronic prostatitis in modern medicine, various theories on the etiology, pathogenesis, and treatment of chronic prostatitis in TCM, entry points from interdisciplinary fields, the role and advantages of TCM in the comprehensive management of chronic prostatitis, and research into mechanisms related to chronic prostatitis. After discussions, the experts concluded that chronic prostatitis is one of the diseases where TCM has a distinct advantage. TCM can effectively improve the clinical syndromes of chronic prostatitis, including urinary issues, social psychology, organ specificity, infection, nervous system problems, pain and discomfort, sexual dysfunction, etc., and can enhance the quality of life. For patients with chronic prostatitis who cannot tolerate or may not accept the adverse effects of Western medicine, as well as for those with non-inflammatory chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), those with pain as the primary symptom, those with urinary tract symptoms, those with coexisting anxiety and depression, and those with abnormal semen liquefaction, TCM-based treatment can be the main approach. Currently, the optimal diagnosis and treatment strategy for chronic prostatitis has not yet been established, and further research from perspectives such as clinical epidemiology, evidence-based medicine, and molecular biology is needed. Both TCM and Western medicine should integrate their respective strengths, clarify the role and advantages of TCM interventions based on mutual reference, and improve the diagnosis and treatment standards for chronic prostatitis in China. 
		                        		
		                        		
		                        		
		                        	
7.Study on Synthesis and Antioxidant Activities in Vitro of Curcumin Pyrazole Derivative
Hua-Jun ZHANG ; Can-Ming LI ; Qin-Xue SUI ; Mei-Qi ZHAN ; Jing GONG ; Li-Ping ZHU ; Tao WANG
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(9):2452-2456
		                        		
		                        			
		                        			Objective To construct curcumin pyrazole derivative by the reaction of diketone of curcumin and benzylhydrazine based on the above structure-activity relationship,and to explore its antioxidant activity to provide experimental basis for the development of curcumin antioxidant derivative.Methods Curcumin-N-substituted pyrazole derivative was synthesized from curcumin and benzylhydrazine.The structures of the derivative were confirmed by infrared spectroscopy(IR),nuclear magnetic resonance spectroscopy(1H-NMR,13C-NMR)and LC-MS.The antioxidant activity in vitro of the derivative was evaluated by determination of curcumin and its pyrazole derivative scavenging ability for 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid(ABTS)free radical.Results Curcumin pyrazole derivative was successfully synthesized.Curcumin and its pyrazole derivative showed good free radical scavenging effects in the range of 4.6-73.6,6.25-100 μg·mL-1,respectively,with a significant dose-effect relationship.The half-maximal inhibition(IC50)values of curcumin and its pyrazole derivatives determined by DPPH method were 14.24,40.37 μg·mL-1,respectively,while the IC50 values of curcumin and its pyrazole derivatives determined by ABTS method were 36.65,19.26 μg·mL-1,respectively.Conclusion The antioxidant activity of β-dione of curcumin was retained through the substitution of the pyrazole ring,and the curcumin pyrazole derivative deserves further investigation as a potential antioxidant.
		                        		
		                        		
		                        		
		                        	
8.Analysis of Ecological Factors and Research Progress in Ecological Planting Mode of Pogostemon cablin
Yun-Zi BAO ; Lin-Yuan CHEN ; Kai-Ying QIU ; Yan-Mei NI ; Han-Qing DING ; Li-Ping WANG ; Zi-Qi LIU ; Ruo-Ting ZHAN ; Li-Kai CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(11):3084-3090
		                        		
		                        			
		                        			Pogostemon cablin is a famous southern medicine.As the important raw material for modern medicine and industry,Pogostemon cablin becomes required with a large marketing demand.However,due to the serious continuous cropping obstacles in the growth process of Pogostemon cablin,the aggravation of diseases of Pogostemon cablin and the degradation of its quality arose.This paper outlined the ecological factors such as climate factors,soil factors and topographic factors suitable for the growth of Pogostemon cablin,analyzed the continuous cropping obstacles and diseases arising in the cultivation,reviewed the current ecological planting mode of Pogostemon cablin such as crop rotation,intercropping,relay-cropping and under-forest planting,and also made a comprehensive evaluation of the economic benefits,ecological benefits and social benefits of the ecological planting mode of Pogostemon cablin,aiming to provide a theoretical basis and a reference for the promotion of the ecological planting mode of Pogostemon cablin.
		                        		
		                        		
		                        		
		                        	
9.Value of multiparametric ultrasonography combined with inflammatory cell ratio in predicting the efficacy of neoadjuvant chemotherapy for breast cancer
Lan GAO ; Yunyun ZHAN ; Jiajia WANG ; Yu BI ; Xiabi WU ; Mei PENG
Chinese Journal of Ultrasonography 2024;33(11):983-991
		                        		
		                        			
		                        			Objective:To investigate the value of multimodal ultrasound features combined with peripheral blood inflammatory cell ratios in evaluating the efficacy of neoadjuvant chemotherapy (NAC) in breast cancer.Methods:A total of 106 breast cancer patients diagnosed and treated with NAC at the Second Affiliated Hospital of Anhui Medical University from May 2021 to April 2024 were retrospectively collected, resulting in the conclusion of 61 patients (61 masses) in the study. All patients underwent multimodal ultrasound and peripheral blood routine examinations before NAC and after two cycles of NAC treatment. The patients were divided into a major histological response (MHR) group and a non-major histological response (NMHR) group as indicators for evaluating NAC efficacy. The differences in multimodal ultrasound features and inflammatory cell ratios before NAC and after two cycles of NAC treatment between the MHR and NMHR groups were compared. Binary logistic multivariate regression analysis was performed to determine the independent predictors of NAC efficacy in breast cancer. ROC curves were plotted to evaluate the diagnostic efficacy of predicting NAC efficacy.Results:Among the 61 breast masses, 25 (40.98%) were in the MHR group, and 36 (59.02%) were in the NMHR group.Multivariate binary logistic regression analysis showed that the change rate of maximum tumor diameter after the second cycle (ΔD 2), change rate of vascular index after the second cycle (ΔVI 2), change rate of elastic strain ratio after the second cycle (ΔE-Strain 2), change rate of reverse imaging score after the second cycle (ΔI-imaging score 2), and platelet-to-lymphocyte ratio (PLR) before NAC were independent predictors of NAC efficacy ( OR=1.145, P=0.019; OR=1.055, P=0.016; OR=1.036, P=0.033; OR=1.276, P=0.016; OR=1.054, P=0.047). The area under the ROC curve (AUC) for the combined diagnosis of the above parameters was 0.928 (95% CI=0.866~0.990), with a sensitivity of 80.0% and a specificity of 91.7%. Conclusions:The combination of ΔD 2, ΔVI 2, ΔE-Strain 2, ΔI-imaging score 2 and PLR before NAC has high clinical application value for early prediction of NAC efficacy in breast cancer.
		                        		
		                        		
		                        		
		                        	
10.The Polymorphism Analysis of HLA Class Ⅱ Alleles Based on Next-Generation Sequencing and Prevention Strategy for Allele Dropout
Su-Qing GAO ; Zhan-Rou QUAN ; Yan-Ping ZHONG ; Hao CHEN ; Liu-Mei HE ; Hong-Yan ZOU ; Zhi-Hui DENG
Journal of Experimental Hematology 2024;32(2):603-609
		                        		
		                        			
		                        			Objective:To investigate the accuracy of next-generation sequencing technology(NGS)in detecting the polymorphisms of HLA-DRB1,DQB1,DQA1,DRB3,DRB4,DRB5,DPA1 and DPB1 alleles in randomly-selected unrelated healthy individuals from Shenzhen Han population,investigate the potential reason for HLA-DRB1 allele dropout in routine NGS,and establish an internal quality control system.Methods:NGS-based HLA class Ⅱ genotyping was performed on 1 012 samples using the MiSeqDxTM platform.The suspected missed alleles indicated by the quality control software and HLA-DRB1 homozygotes were confirmed by PCR-SSOP or PCR-SBT methods.Results:A total of 139 alleles were detected,including HLA-DRB1(45),DRB3(7),DRB4(5),DRB5(7),DQA1(17),DQB1(21),DPA1(10)and DPB1(27).HLA-DRB 1*09:01(17.09%),15:01(10.72%);DRB3*02:02(25.99%),03:01(10.18%);DRB4*01:03(36.46%);DRB5*01:01(15.42%);DQA1*01:02(20.01%),03:02(17.19%);DQB1*03:01(19.47%),03:03(17.98%),05:02(11.66%),06:01(10.67%);DPA1*02:02(54.45%),01:03(31.18%)and DPB1*05:01(39.13%),02:01(16.90%)alleles were the most common alleles in Shenzhen Han population(frequencies>10%).There was no statistical difference between the gene frequencies of HLA-DRB1 and DQB1 loci in our study.The HLA Common and Well-Documented Alleles in China(CWD2.4)(x2=12.68,P>0.05).94 cases of HLA-DRB1 homozygous samples detected by NGS were retested by PCR-SSOP or SBT method,and one case of allele dropout at HLA-DRB1 locus was found.SBT method confirmed that the allele of DRB1*04:03 was missed.The laboratory internal quality control system was established.Two cases of new alleles were detected and named by WHO Nomenclature Committee for Factors of the HLA System.Conclusion:The HLA genotyping results based on NGS showed a significantly lower ambiguity rate.The HLA class Ⅱ alleles exhibit genetic polymorphism in the Han population of unrelated healthy individuals in Shenzhen.The independent method based on NGS in clinical histocompatibility testing has limitations and requires internal quality control strategies to avoid allele-dropout events.
		                        		
		                        		
		                        		
		                        	
            
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