1.Inhibition of M2-type macrophage-mediated migration and epithelial mesenchymal transition in colorectal cancer by bufalin
Donghao TANG ; Jinbao CHEN ; Linlin JIA ; Dongxiao SHEN ; Jing SHANG ; Yuejiao FENG ; Jiahao LU ; Zengyou XIAO ; Yujie HE ; Jie WANG
Acta Universitatis Medicinalis Anhui 2024;59(2):310-315
Objective To investigate the role of bufalin(BU)in inhibiting M2-type macrophage-mediated colorec-tal cancer metastasis.Methods Human acute leukemia mononuclear cells(THP-1)were differentiated into M0 macrophages using phorbol ester induction(PMA)for 48 hours.The M0 macrophages were then treated with IL-4 and IL-13 medium.Surface markers and morphological changes were observed through ELISA,morphology,and RT-qPCR experiments.RT-PCR and ELISA experiments were conducted to detect the surface markers TGF-β and IL-10 of M2 macrophages.The secretion level of IL-6 in the supernatant of M2 macrophages and colorectal cancer cells HCT116 was compared using ELISA.Additionally,the effect of conditioned medium on colorectal cancer cell HCT116 was assessed through Transwell,Wound healing,RT-qPCR,and Western blot experiments.Subsequent-ly,bufalin was added to the conditioned medium and the changes in AKT/PI3K protein,migration,and epithelial-mesenchymal transition ability in HCT116 were observed using Western blot,Transwell,Wound healing and RT-qPCR experiments.Results THP-1 were successfully differentiated into M2 macrophages.The activation of AKT/PI3K protein in HCT116 cells was induced by the secretion of IL-6 from M2 macrophages,which in turn promoted the migration and epithelial-mesenchymal transition ability of the HCT116 cells.The migration and epithelial-mes-enchymal transition mediated by M2 macrophages in HCT116 cells were effectively inhibited by Bufalin.Conclu-sion The release of IL-6 from M2 macrophages activates the AKT/PI3K signaling pathway in colorectal cancer cells,thereby promoting their migration and epithelial-mesenchymal transition capacity.Moreover,bufalin exhibits inhibitory effects on this effect.
2.Adeno-associated virus-mediated hepatocyte-specific NDUFA13 overexpression protects against CCl4-induced liver fibrosis in mice by inhibiting hepatic NLRP3 activation
Xiaohui XU ; Jinmei FENG ; Ying LUO ; Xinyu HE ; Jinbao ZANG ; Daochao HUANG
Journal of Southern Medical University 2024;44(2):201-209
Objective To investigate the protective effect of NDUFA13 protein against acute liver injury and liver fibrosis in mice and explore the possible mechanisms.Methods BALB/C mice(7 to 8 weeks old)were divided into normal group,CCl4 group,CCl4+AAV-NC group and CCl4+AAV-NDU13 group(n=18).Mouse models of liver fibrosis were established by intraperitoneal injection of CCl4 twice a week for 3,5 or 7 weeks,and the recombinant virus AAV8-TBG-NC or AAV8-TBG-NDUFA13 was injected via the tail vein 7-10 days prior to CCl4 injection.After the treatments,pathological changes in the liver of the mice were observed using HE and Masson staining.Hepatic expression levels of NDUFA13 and α-SMA were detected with Western blotting,and the coexpression of NDUFA13 and NLRP3,TNF-α and IL-1β,and α-SMA and collagen Ⅲ was analyzed with immunofluorescence assay.Results HE and Masson staining showed deranged liver architecture,necrotic hepatocytes and obvious inflammatory infiltration and collagen fiber deposition in mice with CCl4 injection(P<0.001).NDUFA13 expression markedly decreased in CCl4-treated mice(P<0.001),while a significant reduction in inflammatory aggregation and fibrosis was observed in mice with AAV-mediated NDUFA13 overexpression(P<0.001).In CCl4+AAV-NDU13 group,immunofluorescence assay revealed markedly weakened activation of NLRP3 inflammasomes(P<0.001),significantly decreased TNF-α and IL-1β secretion(P<0.001),and inhibited hepatic stellate cell activation(P<0.05)and collagen formation in the liver(P<0.001).Conclusion Mitochondrial NDUFA13 overexpression in hepatocytes protects against CCl4-induced liver fibrosis in mice by inhibiting activation of NLRP3 signaling.
3.Adeno-associated virus-mediated hepatocyte-specific NDUFA13 overexpression protects against CCl4-induced liver fibrosis in mice by inhibiting hepatic NLRP3 activation
Xiaohui XU ; Jinmei FENG ; Ying LUO ; Xinyu HE ; Jinbao ZANG ; Daochao HUANG
Journal of Southern Medical University 2024;44(2):201-209
Objective To investigate the protective effect of NDUFA13 protein against acute liver injury and liver fibrosis in mice and explore the possible mechanisms.Methods BALB/C mice(7 to 8 weeks old)were divided into normal group,CCl4 group,CCl4+AAV-NC group and CCl4+AAV-NDU13 group(n=18).Mouse models of liver fibrosis were established by intraperitoneal injection of CCl4 twice a week for 3,5 or 7 weeks,and the recombinant virus AAV8-TBG-NC or AAV8-TBG-NDUFA13 was injected via the tail vein 7-10 days prior to CCl4 injection.After the treatments,pathological changes in the liver of the mice were observed using HE and Masson staining.Hepatic expression levels of NDUFA13 and α-SMA were detected with Western blotting,and the coexpression of NDUFA13 and NLRP3,TNF-α and IL-1β,and α-SMA and collagen Ⅲ was analyzed with immunofluorescence assay.Results HE and Masson staining showed deranged liver architecture,necrotic hepatocytes and obvious inflammatory infiltration and collagen fiber deposition in mice with CCl4 injection(P<0.001).NDUFA13 expression markedly decreased in CCl4-treated mice(P<0.001),while a significant reduction in inflammatory aggregation and fibrosis was observed in mice with AAV-mediated NDUFA13 overexpression(P<0.001).In CCl4+AAV-NDU13 group,immunofluorescence assay revealed markedly weakened activation of NLRP3 inflammasomes(P<0.001),significantly decreased TNF-α and IL-1β secretion(P<0.001),and inhibited hepatic stellate cell activation(P<0.05)and collagen formation in the liver(P<0.001).Conclusion Mitochondrial NDUFA13 overexpression in hepatocytes protects against CCl4-induced liver fibrosis in mice by inhibiting activation of NLRP3 signaling.
4.An interpretable machine learning method for heart beat classification
Jinbao ZHANG ; Peiyu HE ; Pian TIAN ; Jianmin CAI ; Fan PAN ; Yongjun QIAN ; Qijun ZHAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2023;30(02):185-190
Objective To explore the application of Tsetlin Machine (TM) in heart beat classification. Methods TM was used to classify the normal beats, premature ventricular contraction (PVC) and supraventricular premature beats (SPB) in the 2020 data set of China Physiological Signal Challenge. This data set consisted of the single-lead electro-cardiogram data of 10 patients with arrhythmia. One patient with atrial fibrillation was excluded, and finally data of the other 9 patients were included in this study. The classification results were then analyzed. Results The classification results showed that the average recognition accuracy of TM was 84.3%, and the basis of classification could be shown by the bit pattern interpretation diagram. Conclusion TM can explain the classification results when classifying heart beats. The reasonable interpretation of classification results can increase the reliability of the model and facilitate people's review and understanding.
5.KCTD4 interacts with CLIC1 to disrupt calcium homeostasis and promote metastasis in esophageal cancer.
Cancan ZHENG ; Xiaomei YU ; Taoyang XU ; Zhichao LIU ; Zhili JIANG ; Jiaojiao XU ; Jing YANG ; Guogeng ZHANG ; Yan HE ; Han YANG ; Xingyuan SHI ; Zhigang LI ; Jinbao LIU ; Wen Wen XU
Acta Pharmaceutica Sinica B 2023;13(10):4217-4233
Increasing evidences suggest the important role of calcium homeostasis in hallmarks of cancer, but its function and regulatory network in metastasis remain unclear. A comprehensive investigation of key regulators in cancer metastasis is urgently needed. Transcriptome sequencing (RNA-seq) of primary esophageal squamous cell carcinoma (ESCC) and matched metastatic tissues and a series of gain/loss-of-function experiments identified potassium channel tetramerization domain containing 4 (KCTD4) as a driver of cancer metastasis. KCTD4 expression was found upregulated in metastatic ESCC. High KCTD4 expression is associated with poor prognosis in patients with ESCC and contributes to cancer metastasis in vitro and in vivo. Mechanistically, KCTD4 binds to CLIC1 and disrupts its dimerization, thus increasing intracellular Ca2+ level to enhance NFATc1-dependent fibronectin transcription. KCTD4-induced fibronectin secretion activates fibroblasts in a paracrine manner, which in turn promotes cancer cell invasion via MMP24 signaling as positive feedback. Furthermore, a lead compound K279-0738 significantly suppresses cancer metastasis by targeting the KCTD4‒CLIC1 interaction, providing a potential therapeutic strategy. Taken together, our study not only uncovers KCTD4 as a regulator of calcium homeostasis, but also reveals KCTD4/CLIC1-Ca2+-NFATc1-fibronectin signaling as a novel mechanism of cancer metastasis. These findings validate KCTD4 as a potential prognostic biomarker and therapeutic target for ESCC.
6.A new method for high-density surface electromyography decomposition in dynamic muscle contraction.
Jinbao HE ; Binglei GUAN ; Kai HUANG ; Zaifei LUO
Journal of Biomedical Engineering 2021;38(6):1081-1086
In this paper, a new surface electromyography (sEMG) signal decomposition method based on spatial location is proposed for the high-density sEMG signals in dynamic muscle contraction. Firstly, according to the waveform correlation of each muscle motor units (MU) in each channel, the firing times are extracted, and then the firing times are classified by the spatial location of MU. The MU firing trains are finally obtained. The simulation results show that the accuracy rate of a single MU firing train after classification is more than 91.67%. For real sEMG signals, the accuracy rate to find a same MU by the "two source" method is over (88.3 ± 2.1)%. This paper provides a new idea for dynamic sEMG signal decomposition.
Action Potentials
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Computer Simulation
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Electromyography
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Muscle Contraction
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Muscle, Skeletal
7. Application of valvuloplasty in mitral valve insufficiency in infants and small children
Siyi HE ; Jinbao ZHANG ; Dong XIN ; Fan WU ; Xiaochen WU ; Feng GAO ; Hui OUYANG
Chinese Journal of Applied Clinical Pediatrics 2019;34(22):1725-1727
Objective:
To explore the efficacy of valvuloplasty on mitral valve regurgitation (MR) in infants and small children, in order to provide evidence for clinical treatment.
Methods:
Retrospective analysis was conducted in patients aged less than 3 years old who have undergone mitral repair surgeries at Department of Cardiovascular Surgery, General Hospital of Western Theater Command from January 2015 to December 2017.Cardiopulmonary bypass (CPB) surgery was performed under general anesthesia.The corresponding repair operations were selected for various MR types.All the children were examined by cardiac ultrasound before discharge.Patients were followed up after surgery and mitral regurgitation was recorded.
Results:
A total of 69 patients were enrolled, including 30 boys and 39 girls, with an average age of (19.3±11.6) months and an average weight of (9.6±2.1) kg.There were 32 cases of Carpentier type Ⅰ, 30 cases of type Ⅱ and 7 cases of type Ⅲ.Twenty-nine patients were diagnosed as simple MR, while the other 40 patients were combined with other cardiac malformations.One patient died at postoperative day 7.One patient still had severe MR after surgery, and then second operation was performed on the 10th day after first surgery.Sixty-five patients were followed up from 1 month to 3 years[(15.9±10.9) months]. During the follow-up period, no patient died or had reoperation.Aggravated MR was observed in 4 patients.
Conclusions
Mitral valvuloplasty is a safe and effective surgical method for treating infants and younger children with MR.The key to the success of surgery is to choose the appropriate individualized repair operation.At the same time, the growth potential of mitral valve should be fully considered to avoid restrained development.
8.Application of Remote Sensing Technology in Investigating Planting Area of Paeoniaceae suffruticosa in Shicheng County of Jiangxi Province
Huiting ZENG ; Xiaoqun HE ; Chao CHEN ; Miaoting CAI ; Yuanjian YUAN ; Xingxing CHEN ; Jinbao YU ; Xiaoqing WANG
China Pharmacy 2019;30(24):3404-3407
OBJECTIVE: To provide reference for the development and sustainable utilization of TCM industry with regional characteristics. METHODS: Taking Shicheng county of Jiangxi province as an example, field investigation was carried out on Paeoniaceae suffruticosa planting base in the county, a few representative P. suffruticosa planting bases in the county were selected as sample points, and GPS was used to locate and record the location information of sample points. The remote sensing image was automatically extracted by computer, the artificial visual interpretation method was used to get P. suffruticosa planting area image. Then combined with the field inspection verification, P. suffruticosa planting area was obtained, and the investigation results were analyzed. RESULTS: Through remote sensing interpretation of the planting area of P. suffruticosa in Shicheng county, it was obtained that the total planting area of P. suffruticosa in Shicheng county was 42 597 951.505 square meters (63 864.995 mu) in 2018, accounting for about 33% of the cultivated land area, which was 42.12% higher than the conventional planting area of 44 936 mu in 2013. The distribution of P. suffruticosa planting in Shicheng county was mainly concentrated in Xiaosong town and Fengshan town in the north, and Daqu town and Pingshan town in the south. CONCLUSIONS: Remote sensing technology has the advantages of fast data acquisition, large amount of information, high accuracy and strong timeliness, which greatly avoids the complexity of work, saves a lot of manpower and material resources. The technology can provide technology support for obtaining the regional planting area and distribution information of TCM such as P. suffruticosa, dynamic monitoring, scientific warning of the market status of TCM, and guiding the large-scale, standardized and intensive development of TCM cultivation.
9.Separation and Determination of Rhynchophylline and Isorhynchophylline in Uncaria rhynchophylla by HPLC with Ionic Liquid as Mobile Phase Additives
Yuanyuan ZHANG ; Huiting ZENG ; Chao CHEN ; Xiaoqun HE ; Yanzhen HU ; Le CHEN ; Jinbao YU
China Pharmacy 2019;30(21):2952-2957
OBJECTIVE: To establish a method to determine the contents of rhynchophylline and isorhynchophylline in Uncaria rhynchophylla. METHODS: The separation degree of ionic liquid 1-butyl-3-methylimidazolium chloride (C4mimCl) as mobile phase additive was compared with that of mobile phase without additives and with traditional additive triethylamine (which damaged the chromatographic column). The optimum concentration of C4mimCl was screened and the contents of rhynchophylline and isorhynchophylline in U. rhynchophylla from 4 habitats in Jiangxi province were determined by the newly established method. The determination was performed on Dikmatech Diamonsil Plus C18 column, the mobil phase was acetonitrile-buffer (0.1% phosphoric acid+3.0 mmol/L C4mimCl), gradient elution. UV detection wavelength was set at 245 nm and the flow rate was 1 mL/min. Sample size was 10 μL. RESULTS: When mobile phase had no additives or 3.0 mmol/L triethylamine and 3.0 mmol/L C4mimCl were added as additives, the separation of rhynchophylline from the front peak was 1.02, 1.23 and 1.72, and the separation from the back peak was 1.06, 6.00 and 4.25, respectively. The symmetry factors were 0.81, 0.86 and 1.13, respectively. The separation of isorhynchophylline from the front peak was 0.96, 3.89 and 4.05, and the separation from the back peak was 1.02, 2.34 and 2.36, respectively. The symmetry factors were 0.88, 0.81 and 0.96, respectively. The linear range of rhynchophylline and isorhynchophylline were 4.93-157.76 (r=0.999 9) and 4.98-159.50 μg/mL (r=1.000), respectively. The quantitative limits were 0.486 4, 0.793 6 μg/mL, respectively. RSDs of precision, repeatability, stability and durability tests were all less than 5% (n=6). The recovery rates were 102.9%-107.8% (RSD=1.7%,n=6) and 95.4%-106.3% (RSD=3.9%,n=6), respectively. The content of rhynchophylline and isorhynchophylline in U. rhynchophylla from 4 habitats were 0.758-1.343 and 1.511-1.823 mg/g, respectively. CONCLUSIONS: Addition of C4mimCl into mobile phase can enhance its separation. Established HPLC method is rapid, accurate and reproducible, which can be used for content determination of rhynchophylline and isorhynchophylline in U. rhynchophylla.

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