1.Effect of rotating cell culture system on proliferation and stemness maintenance of epidermal stem cells in the elderly
Min ZHANG ; Xixi DONG ; Meng HUANG ; Chaoxiang WANG ; Luyue ZHANG ; Junkai CAO
Chinese Journal of Tissue Engineering Research 2025;29(13):2698-2705
BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.
2.Prediction of Expression of Ki-67 Status in Breast Cancer via Deep Learning-Based Radiomics Model
Hanmin XIE ; Jialing CHENG ; Yuelong LI ; Chengwei LI ; Chaoxiang YANG ; Ruoxian ZHANG
Chinese Journal of Medical Imaging 2025;33(10):1049-1055
Purpose To analyze the value of a deep learning(DL)radiomics model based on dynamic contrast-enhanced MRI images in predicting the expression of Ki-67 status in breast cancer.Materials and Methods A retrospective analysis of 152 breast cancer patients confirmed by pathological results at Guangdong Women and Children Hospital,MRI images and clinical pathological data were reviewed,and based on postoperative immunohistochemistry results,the images of the high and low expression groups of Ki-67 were randomly sampled in a ratio of 8∶2 to form a training set of 122 cases and a validation set of 30 cases.Single-factor and multi-factor Logistic regression analyses of clinical data were performed to select independent predictors of breast cancer expressing Ki-67 status.The ResNet-18 model was used as the basic model for DL feature extraction.Hand-crafted radiomic features and DL features were extracted.Eight machine learning models were constructed based on clinical features,hand-crafted radiomic features,DL features,and their combinations.The area under the receiver operating characteristic curve was used to evaluate the predictive performance of the models,and the best model was determined as the output model.Results The progesterone receptor status(OR=0.764,P=0.040)and human epidermal growth factor receptor-2 status(OR=1.187,P=0.046)were independent clinical predictors of breast cancer expressing Ki-67 status.The combined feature models demonstrated superior performance over the individual feature models,and the support vector machine algorithm had the highest prediction performance in the validation set,with an area under the curve of 0.847.Conclusion The DL radiomics model based on dynamic contrast-enhanced MRI images can effectively predict the expression of Ki-67 status in breast cancer.The support vector machine algorithm combined with feature model is the best,which can help the clinical diagnosis and treatment of breast cancer and prognosis evaluation.
3.Prediction of Expression of Ki-67 Status in Breast Cancer via Deep Learning-Based Radiomics Model
Hanmin XIE ; Jialing CHENG ; Yuelong LI ; Chengwei LI ; Chaoxiang YANG ; Ruoxian ZHANG
Chinese Journal of Medical Imaging 2025;33(10):1049-1055
Purpose To analyze the value of a deep learning(DL)radiomics model based on dynamic contrast-enhanced MRI images in predicting the expression of Ki-67 status in breast cancer.Materials and Methods A retrospective analysis of 152 breast cancer patients confirmed by pathological results at Guangdong Women and Children Hospital,MRI images and clinical pathological data were reviewed,and based on postoperative immunohistochemistry results,the images of the high and low expression groups of Ki-67 were randomly sampled in a ratio of 8∶2 to form a training set of 122 cases and a validation set of 30 cases.Single-factor and multi-factor Logistic regression analyses of clinical data were performed to select independent predictors of breast cancer expressing Ki-67 status.The ResNet-18 model was used as the basic model for DL feature extraction.Hand-crafted radiomic features and DL features were extracted.Eight machine learning models were constructed based on clinical features,hand-crafted radiomic features,DL features,and their combinations.The area under the receiver operating characteristic curve was used to evaluate the predictive performance of the models,and the best model was determined as the output model.Results The progesterone receptor status(OR=0.764,P=0.040)and human epidermal growth factor receptor-2 status(OR=1.187,P=0.046)were independent clinical predictors of breast cancer expressing Ki-67 status.The combined feature models demonstrated superior performance over the individual feature models,and the support vector machine algorithm had the highest prediction performance in the validation set,with an area under the curve of 0.847.Conclusion The DL radiomics model based on dynamic contrast-enhanced MRI images can effectively predict the expression of Ki-67 status in breast cancer.The support vector machine algorithm combined with feature model is the best,which can help the clinical diagnosis and treatment of breast cancer and prognosis evaluation.
4.Effect of rotating cell culture system on proliferation and stemness maintenance of epidermal stem cells in the elderly
Min ZHANG ; Xixi DONG ; Meng HUANG ; Chaoxiang WANG ; Luyue ZHANG ; Junkai CAO
Chinese Journal of Tissue Engineering Research 2025;29(13):2698-2705
BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.
5.Analysis of prenatal ultrasound signs and prevention strategies for missed diagnosis and misdiagnosis in Apert syndrome
Huijuan LIN ; Limin WANG ; Li ZHENG ; Chaoxiang YANG ; Ning SHANG ; Zhen XIAO ; Fangfang XU
Chinese Journal of Ultrasonography 2024;33(9):784-790
Objective:To summarize the key prenatal ultrasound diagnosis features of Apert syndrome, analyze the causes of missed diagnosis and misdiagnosis, and propose corresponding preventive strategies.Methods:A retrospective analysis was made on the medical records and prenatal ultrasound images of 15 fetuses (including 14 cases referred from other hospitals) who underwent prenatal ultrasound examination in Guangdong Women and Children Hospital from August 2014 to May 2022 and were eventually clinically confirmed as Apert syndrome by induction or after birth. By conducting a comparative analysis, particularly focusing on the initial and final diagnoses of referral cases, the key ultrasound diagnostic points of Apert syndrome and the causes for missed and misdiagnosis were summarized.Results:①Diagnostic accuracy: Among the 15 fetuses, 11 cases (73.3%) were correctly diagnosed by prenatal ultrasound and 4 cases (26.7%) were missed diagnosis and misdiagnosis. For the 14 referral cases, only 2 cases (14.3%) were correctly identified in the initial diagnosis at the referring hospital (14.3%), and 12 cases (85.7%) were missed missed diagnosis and misdiagnosis. ②Detection rate of ultrasound signs: In the 15 fetuses with Apert syndrome, the detection rate of " cloverleaf" skull was 13.3% (2/15), premature coronal suture was 66.7% (10/15), the " brain shadowing sign" and flat occiput were both 93.3% (14/15), prominent forehead, hypertelorism and bilateral syndactyly of hands were all 100% (15/15), and bilateral syndactyly of feet was 73.3% (11/15). ③Analysis of missed diagnosis and misdiagnosis: Among the 4 cases of missed diagnosis and misdiagnosis in our hospital, premature closure of coronal suture, " brain shadowing sign", flat occiput and hypertelorism were all not recognized. Among these, 3 cases also missed the prominent forehead, bilateral syndactyly of hands and feet. Additionally, 1 case of bilateral syndactyly of hands was misdiagnosed as partial absence of metacarpals and phalangess.Conclusions:In the prenatal ultrasound diagnosis of fetal Apert syndrome, the symmetric syndactyly of both hands serves as an extremely important diagnostic clue. The " cloverleaf" skull is not common. The premature closure of coronal suture as a direct diagnostic sign with a high detection rate, highlighting its significance in the diagnostic of Apert syndrome. Furthermore, the high detection rates of characteristic ultrasound features such as prominent forehead, flat occiput, " brain shadowing sign" and hypertelorism could help to improve the accuracy of prenatal ultrasound diagnosis for Apert syndrome and effectively reduce missed and misdiagnosis.
6.A study of longitudinal trajectories and predictive factors of symptom clusters in patients with laparoscopic anal preservation surgery for rectal cancer
Chaoxiang YOU ; Mengying JIA ; Shuang LI ; Lili CHEN ; Wendan JING ; Hongyan KOU
Chinese Journal of Nursing 2024;59(8):922-929
Objective To explore the longitudinal trajectories of symptom clusters in patients with rectal cancer within 6 months after laparoscopic anal preservation surgery,and analyze the predictive factors of each trajectory subgroup.Methods A longitudinal survey was conducted to select 128 patients who underwent laparoscopic anal preservation surgery for rectal cancer at the Department of Gastrointestinal Surgery,a tertiary hospital in Nanchong from November 2021 to April 2022.The general information questionnaire,Chinese version of Anderson Symptom Inventory for Gastrointestinal Cancer and Charlson Comorbidity Index were used to conduct follow-up surveys of the selected patients at 2 weeks,1 month,3 months and 6 months after surgeiy.The symptom cluster were extracted by exploratory factor analysis,and the latent category growth model was constructed to identify the trajectory subgroups of each symptom cluster.The predictive factors of each trajectory subgroup were analyzed by Mplus statistical software.Results There were 4 symptom clusters within 6 months after laparoscopic anus preserving surgeiy for rectal cancer,which were named as sickness symptom cluster,psychological-sleep symptom cluster,gastrointestinal symptom cluster,and psychological-treatment-related symptom cluster.The variance contribution rates were 65.173%,66.225%,62.421%,and 60.492%,respectively.The latent class growth model identified that there are 4 to 5 trajectory subgroups in the symptom cluster.Gender,education level,Charles Comorbidity Index,clinical T stage,medical cost burden,and treatment method were the predictors of high-risk trajectory subgroups in the symptom cluster(P<0.05).Conclusion There are 4 symptom clusters within 6 months after laparoscopic anus preserving surgeiy for rectal cancer.The gastrointestinal symptom cluster is a specific symptom cluster for this type of patients with the heaviest symptom burden.The overall trajectory of each symptom cluster shows a dynamic decline trend,but individual trajectoiy subgroups still have an exacerbation trend.There are differences in predictive factors for different symptom clusters.Clinical medical staff should pay attention to the management of symptom clusters in patients with female patients,low education level,high Charles Comorbidity Index,high clinical T stage,heavy medical cost burden,and patients whose treatment method is chemotherapy plus surgery.High-risk groups should be timely identified;corresponding intervention methods were dynamically adjusted;the quality of precision nursing was improved.
7.Cloning, expression and activity analysises of chalcone synthase genes in Carthamus tinctorius.
Xiaohui TANG ; Chaoxiang REN ; Jing HU ; Jiang CHEN ; Jie WANG ; Rui WANG ; Qinghua WU ; Wan LIAO ; Jin PEI
Chinese Herbal Medicines 2023;15(2):291-297
OBJECTIVE:
Flavonoids are the bioactive compounds in safflower (Carthamus tinctorius), in which chalcone synthase (CHS) is the first limiting enzyme. However, it is unclear that which chalcone synthase genes (CHSs) are participated in flavonoids biosynthesis in C. tinctorius. In this study, the CHSs in the molecular characterization and enzyme activities were investigated.
METHODS:
Putative chalcone biosynthase genes were screened by the full-length transcriptome sequences data in C. tinctorius. Chalcone biosynthase genes in C. tinctorius (CtCHSs) were cloned from cDNA of flowers of C. tinctorius. The cloned gene sequences were analyzed by bioinformatics, and their expression patterns were analyzed by real-time PCR (RT-PCR). The protein of CtCHS in the development of flowers was detected by polyclonal antibody Western blot. A recombinant vector of CtCHS was constructed. The CtCHS recombinant protein was induced and purified to detect the enzyme reaction (catalyzing the reaction of p-coumaryl-CoA and malonyl-CoA to produce naringin chalcone). The reaction product was detected by HPLC and LC-MS.
RESULTS:
Two full-length CtCHS genes were successfully cloned from the flowers of safflower (CtCHS1 and CtCHS3), with gene lengths of 1525 bp and 1358 bp, respectively. RT-PCR analysis showed that both genes were highly expressed in the flowers, but the expression of CtCHS1 was higher than that of CtCHS3 at each developmental stage of the flowers. WB analysis showed that only CtCHS1 protein could be detected at each developmental stage of the flowers. HPLC and LC-MS analyses showed that CtCHS1 could catalyze the conversion of p-coumaryl-CoA and malonyl-CoA substrates to naringin chalcone.
CONCLUSION
CtCHS1 is involved in the biosynthesis of naringin chalcone in safflower.
8.Prenatal diagnosis and intrauterine treatment of a giant fetal hepatic hemangioma:a case report
Hui TANG ; Chaoxiang YANG ; Jingshu LI ; Wei WANG ; Dan CHEN ; Jing WU
Chinese Journal of Perinatal Medicine 2023;26(4):331-334
This article reported a case of fetal giant hepatic hemangioma with cardiomegaly managed with intrauterine treatment. At 23 weeks of gestation, the patient was referred to Guangdong Women and Children Hospital due to abnormal abdominal echogenicity of the fetus, which was suspected to be a hepatic hemangioma or a hepatic arteriovenous fistula. The prenatal ultrasound at 26 weeks of gestation revealed an enlarged fetal hepatic hemangioma of 45 mm×35 mm×42 mm and an enlarged heart (cardiothoracic area ratio of 0.50). So, with the patient's informed consent, the fetus was treated with intrauterine administration of propranolol and dexamethasone and closely monitored by ultrasound. The volume of the lump still increased at the beginning of the medication, but started to shrink in the 7th week. Besides, the fetal cardiac load was reduced and the condition was controlled. The patient delivered at 37 weeks of gestation. The baby received a CT examination on the fourth day after birth which revealed an abdominal mass of 40 mm×30 mm×44 mm requiring no treatment, and no abnormalities were reported during a one-year follow-up.
9.Self-compassion in patients after colorectal cancer surgery:current situations and influencing factors
Shuang LI ; Mengying JIA ; Chaoxiang YOU ; Lili CHEN ; Hongyan KOU ; Shoujiang WEI
Modern Clinical Nursing 2023;22(10):8-15
Objective To investigate the current situations and analyse the influencing factors of self-compassion in patients 3 months after colorectal cancer surgery,so as to provide a basis for clinical interventions.Methods This cross-sectional survey was conducted from August 2020 to January 2022 among 190 patients who were at 3 months after colorectal cancer surgery in a tertiary hospital in sichuan.General information questionnaire,self-compassion scale,Connor-Davidson resilience scale and meaning in life questionnaire were used in the survey.Results Total scores of self-compassion,mental toughness and meaning of life after colorectal cancer surgery were 76.69±9.68,58.01±12.54 and 36.02±12.22,respectively.With analysis of multiple linear regression,the influencing factors of self-compassion were found as comorbid chronic diseases,postoperative stoma,postoperative radiotherapy,mental toughness and meaning of life(all P<0.05),toally explaining 58.4%its varience.Conclusions The self-compassion of patients with colorectal cancer is at a moderately lower level and the influencing factors of it include comorbid chronic diseases,postoperative stoma,postoperative radiotherapy,mental toughness and meaning of life.Medical staff should assess the psychological status of the patients,strengthen a psychological care and help patients to gain correct cognitive views for the purpose to improve the self-compassion of patients.
10. Effects of miR-195-5p on ox-LDL-induced vascular endothelial cell injury through regulating FBXW7
Naping LIN ; Yutang WU ; Defen HUANG ; Chaoxiang XU
Chinese Journal of Clinical Pharmacology and Therapeutics 2022;27(2):163-170
AIM: To investigate the effect of miR-195-5p on ox-LDL-induced vascular endothelial cell injury and its mechanism. METHODS: The expression of miR-195-5p in HUVECs cells induced by ox-LDL was detected by RT-PCR. Cell proliferation, the expression of oxidative stress associated factors (MDA and LDH) and apoptosis were detected by CCK-8, ELISA and flow cytometry, respectively. The potential targets of miR-195-5p were determined by dual luciferase reporter assay. The expression of target protein in ox-LDL-induced HUVECs cells and the relationship between miR-195-5p and FBXW7 expression were detected by Western blotting. RESULTS: The expression of miR-195-5p in ox-LDL-induced HUVECs cells was significantly higher than that in normal HUVECs cells. Subsequently, miR-195-5p silencing in ox-LDL-induced HUVECs cells effectively promoted the proliferation of HUVECs cells, whereas suppressed the expression of oxidative stress associated factors (MDA and LDH) and apoptosis level. Luciferase reporter assay and Western blot results showed that miR-195-5p could directly target FBXW7 protein gene and negatively regulate its expression. Finally, miR-195-5p modulated the proliferation, oxidative stress and apoptosis of HUVECs cells induced by ox-LDL by regulating the expression of FBXW7. CONCLUSION: miR-195-5p is highly expressed in HUVECs cells induced by ox-LDL. In addition, miR-195-5p can aggravate ox-LDL-induced cytotoxicity, oxidative stress and apoptosis of HUVECs cells by inhibiting the expression of FBXW7.

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