1.Preparation and characterization of 3D plant-based scaffold based on decellularization method in liver tissue engineering
Jingjing HU ; Songlin HE ; Daxu ZHANG ; Shuo ZHAO ; Xiaonan SHI ; Weilong LI ; Shujun YE ; Jingyi WANG ; Quanyi GUO ; Li YAN
Chinese Journal of Tissue Engineering Research 2024;28(29):4645-4651
BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.
2.A survey on the intelligence status of children born during iodine deficiency disorders intervention period in rural areas of Linxia Hui Autonomous Prefecture, Gansu Province
Aiwei HE ; Limei YIN ; Yanling WANG ; Qinglin LI ; Xiaonan ZHU ; Xiulan FEI ; Jing ZHENG ; Yongqin CAO
Chinese Journal of Endemiology 2024;43(3):229-232
Objective:To investigate the intelligence status of children aged 10 - 12 in rural areas of Linxia Hui Autonomous Prefecture (referred to as Linxia) in Gansu Province.Methods:From September to November 2019, a cross-sectional study was carried out to investigate the intelligence status of children aged 10 to 12 in 8 counties (cities) of Linxia. Chinese Raven's Progressive Matrices (rural version) was used for intelligence test and children's intelligence quotient (IQ) was calculated by regular mold to evaluate children's intelligence level.Results:A total of 1 721 children in Linxia were tested for intelligence, with an average IQ of 103.2. Among them, low intelligence (≤69) accounted for 1.0% (18/1 721), borderline (70 - 79) accounted for 3.0% (52/1 721), middle and lower (80 - 89) accounted for 8.4% (144/1 721), moderate (90 - 109) accounted for 56.6% (974/1 721), middle and upper (110 - 119) accounted for 21.9% (377/1 721), excellent (120 - 129) accounted for 7.8% (135/1 721), extremely excellent (≥130) accounted for 1.2% (21/1 721). There were 635, 598 and 488 children aged 10, 11 and 12, respectively, with an average IQ of 106.1, 103.3 and 99.2. There were 919 males and 802 females, with an average IQ of 102.9 and 103.4, respectively.Conclusion:In 2019, the intelligence of children aged 10 to 12 in rural areas of Linxia has reached the moderate level.
3.Safety of X-ray-versus ultrasound-guided femoral artery puncture in elderly patients:an analysis based on propensity score matching
He YAN ; Dongyan ZHANG ; Xu GUO ; Yuqing GUO ; Ning MA ; Jianjun ZHANG ; Xiaonan GUAN
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(7):774-778
Objective To assess the safety of femoral artery puncture procedures guided by X-ray and ultrasound among elderly patients.Methods A total of 480 patients undergoing transcatheter interventional treatment for cardiovascular and cerebrovascular diseases through the femoral ar-tery in our hospital between January 2016 and December 2022 were enrolled in the study.Of them,326 patients receiving femoral artery puncture guided by X-ray fluoroscopy were assigned into X-ray group,while the other 154 patients guided by vascular Doppler ultrasound were into ultrasound group.With propensity score matching(PSM)in a ratio of 1∶1,finally 270 patients were included.Their general clinical data,success rate of puncture,puncture site,and incidence of vascular complications were compared between the two groups.Multivariate logistic regression analysis was used to identify the risk factors for vascular complications.Results Before PSM,there were no statistical differences in the mean distance from the skin fold to the bifurcation of the common femoral artery(2.5±1.0 cm vs 2.4±0.8 cm)or the distance from the fold to the in-guinal ligament(6.4±1.4 cm vs 6.3±1.7 cm)between the X-ray group and the ultrasound group(P>0.05).After PSM,the X-ray group exhibited an obviously higher incidence of puncture points below the common femoral artery than the ultrasound group(14.8%vs 6.7%,P<0.05),but no significant differences were observed in the one-time success rate of puncture or the occur-rence of vascular complications between the two groups(P>0.05).Multivariate logistic regres-sion analysis indicated that the presences of non-common femoral artery and femoral artery calci-fication at the puncture site was independent risk factors for vascular complications(OR=8.379,95%CI:3.561-19.717;OR=3.922,95%CI:1.664-9.242).Conclusion There is no statistical disparity in safety between X-ray-versus ultrasound-guided femoral artery puncture procedures.Cli-nicians should choose appropriate puncture procedure or combine them together based on individual con-dition of patients.
4.The preliminary application of cinematic rendering reconstruction technology in acute aortic dissection
He ZHANG ; Zhongxiao LIU ; Meng YU ; Miao YU ; Ziyou WANG ; Wenbei XU ; Xiaonan SUN ; Shenman QIU ; Lixiang XIE ; Yanchun ZHANG ; Yankai MENG ; Cunjie SUN ; Kai XU
Journal of Practical Radiology 2024;40(10):1620-1624
Objective To analyze the clinical application value of cinematic rendering(CR)reconstruction technology in acute aortic dissection(AAD),and to compare the imaging quality between CR and volume rendering(VR)reconstruction.Methods Patients with suspected A AD who underwent aortic computed tomography angiography(CTA)were analyzed retrospectively.All images were uploaded to Siemens Syngo.via post-processing workstation for VR and CR three-dimensional reconstruction,respectively.The optimized view angle,staining and transparency were selected and segmented by a radiologist to display the lesion to the full extent.All subjective evaluations of post-processing images were randomly evaluated on Siemens Syngo.via post-processing workstation by two radiologists.The two radiologists reached a consensus after consultation,and the results without consensus were evaluated by another senior radiologist.The 3-point scale was used in the subjective evaluation of post-processing images.The scores of rupture,endometrium,and true and false cavity were recorded.The diagnostic confidence was also recorded.Results A total of 21 ADD patients were enrolled,11 patients(52.3%)were Debakey Ⅲ type.The scores of rupture in CR and VR reconstruction were 2.952 points and 2.619 points,respectively,which had significant difference(P=0.016).For the endometrium of AAD,the score of all 21 patients in the CR reconstruction was 3 points,while only 7 patients(33.3%)in the VR reconstruction had 3 points,which showed significant difference between the both(P<0.001).For the true and false cavity of AAD,only 1 patient(4.8%)in the VR reconstruction was 3 points,while all 21 patients in the CR reconstruction had 3 points(P<0.001).The scores of CR reconstruction on the diagnostic confidence were significantly higher than those of VR reconstruction(P<0.001).Conclusion CR reconstruction can provide photorealistic anatomical post-processing images,and can improve the display and evaluation of AAD.
5.Correlation between CT attenuation value of pulmonary artery thrombi and efficacy of interventional thrombolysis in patients with acute pulmonary embolism
Xiaonan SUN ; Zhongxiao LIU ; He ZHANG ; Xin TANG ; Shenman QIU ; Yankai MENG ; Lixiang XIE ; Shaodong LI ; Qingqiao ZHANG ; Kai XU
Chinese Journal of General Practitioners 2024;23(7):728-733
Objective:To analyze the relationship between CT attenuation value of pulmonary artery thrombi and the efficacy of interventional thrombolysis in patients with acute pulmonary embolism (APE).Methods:This was a single center cross-sectional study. The clinical and imaging data of 89 APE patients who underwent interventional thrombolysis in Affiliated Hospital of Xuzhou Medical University from January 2018 to December 2022, were retrospectively analyzed. All patients underwent CT pulmonary angiography (CTPA) before and after thrombolysis, the CT attenuation value of pulmonary artery thrombi and ratio of CT attenuation value of thrombi to left subscapularis muscle CT value were obtained; and the difference of Qanadli embolism index (ΔQ) before and after thrombolysis was calculated. According to the median ΔQ, patients were classified as good efficacy group (ΔQ>50%) and poor efficacy group (ΔQ≤50%). The clinical characteristics and quantitative parameters of CT were compared between the two groups, and the factors associated with efficacy of thrombolysis were analyzed with univariate and multivariate logistic regression. The correlation between CT attenuation value of pulmonary artery thrombi and ΔQ was analyzed by Spearman correlation analysis.Results:The CT attenuation value of thrombi and ratio of attenuation value of thrombi to left subscapularis muscle CT value showed significant difference between the two groups ( P<0.05). Multivariate analysis showed that compared with CT attenuation value of emboli≤53.47 HU, the value>53.47 HU might be associated with the good efficacy of thrombosysis ( OR=9.175, 95% CI: 0.937-89.846, P=0.057). There was a positive correlation between CT value of pulmonary artery thrombi and ΔQ ( r=0.365, P<0.001). Conclusion:The CT attenuation value of thrombi can predict the efficacy of interventional thrombolysis in APE patients, and patients with higher CT attenuation value would have a better treatment response.
6.Maackiain inhibits proliferation and promotes apoptosis of nasopharyngeal carcinoma cells by inhibiting the MAPK/Ras signaling pathway.
Xing JIANG ; Xiaonan YANG ; Yanxia SHI ; Yan LONG ; Wenqing SU ; Wendong HE ; Kunhua WEI ; Jianhua MIAO
Chinese Journal of Natural Medicines (English Ed.) 2023;21(3):185-196
Nasopharyngeal carcinoma (NPC) is the third most common malignancy with a high recurrence and metastasis rate in South China. Natural compounds extracted from traditional Chinese herbal medicines have been developed and utilized for the treatment of a variety of cancers with modest properties and slight side effects. Maackiain (MA) is a type of flavonoid that was first isolated from leguminous plants, and it has been reported to relieve various nervous system disorders and exert anti-allergic as well as anti-inflammatory effects. In this study, we demonstrated that MA inhibited proliferation, arrested cell cycle and induced apoptosis in nasopharyngeal carcinoma CNE1 and CNE2 cells in vitro and in vivo. The expression of the related proteins associated with these processes were consistent with the above effects. Moreover, transcriptome sequencing and subsequent Western blot experiments revealed that inhibition of the MAPK/Ras pathway may be responsible to the anti-tumor effect of MA on NPC cells. Therefore, the effects of MA and an activator of this pathway, tertiary butylhydroquinone (TBHQ), alone or combination, were investigated. The results showed TBHQ neutralized the inhibitory effects of MA. These data suggest that MA exerts its anti-tumor effect by inhibiting the MAPK/Ras signaling pathway and it has the potential to become a treatment for patients with NPC.
Humans
;
Nasopharyngeal Carcinoma/pathology*
;
Cell Line, Tumor
;
Cell Proliferation
;
Apoptosis
;
Signal Transduction
;
Nasopharyngeal Neoplasms/pathology*
7.Role of ceRNA network in inflammatory cells of rheumatoid arthritis.
Xiaoyu HE ; Haohua HE ; Yan ZHANG ; Tianyu WU ; Yongjie CHEN ; Chengzhi TANG ; Tian XIA ; Xiaonan ZHANG ; Changhao XIE
Journal of Central South University(Medical Sciences) 2023;48(5):750-759
Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease caused by inflammatory cells. Various inflammatory cells involved in RA include fibroblast-like synoviocytes, macrophages, CD4+T-lymphocytes, B lymphocytes, osteoclasts and chondrocytes. The close interaction between various inflammatory cells leads to imbalance of immune response and disorder of the expression of mRNA in inflammatory cells. It helps to drive production of pro-inflammatory cytokines and stimulate specific antigen-specific T- and B-lymphocytes to produce autoantibodies which is an important pathogenic factor for RA. Competing endogenous RNA (ceRNA) can regulate the expression of mRNA by competitively binding to miRNA. The related ceRNA network is a new regulatory mechanism for RNA interaction. It has been found to be involved in the regulation of abnormal biological processes such as proliferation, apoptosis, invasion and release of inflammatory factors of RA inflammatory cells. Understanding the ceRNA network in 6 kinds of RA common inflammatory cells provides a new idea for further elucidating the pathogenesis of RA, and provides a theoretical basis for the discovery of new biomarkers and effective therapeutic targets.
Humans
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Arthritis, Rheumatoid/genetics*
;
MicroRNAs/metabolism*
;
Synoviocytes/pathology*
;
Cytokines/metabolism*
;
RNA, Messenger/metabolism*
;
Fibroblasts/pathology*
;
Cell Proliferation
8.Inhibitory Effect and Target Prediction of Genistein on the Growth of Human Nasopharyngeal Carcinoma CNE 1 Cells
Wendong HE ; Wenqing SU ; Kunhua WEI ; Ling KUI ; Shuo WANG ; Xiaomei GONG ; Xiaonan YANG ; Jianhua MIAO
China Pharmacy 2021;32(10):1196-1204
OBJECTIVE:To study the inhibi tory effects of genistein on the growth of human nasopharyngeal carcinoma. CNE 1 cells and predict its potential target. METHODS :CCK-8 method was used to test the effects of 0(blank control ),12.5,25,50, 100,150 µmol/L genistein on the proliferation of CNE 1 cells after treated for 24,48,72 h. Flow cytometry was carried out to detect the effects of 0(blank control ),15,30,60 µmol/L genistein on the cell cycle and ap optosis of CNE 1 cells after treated for 24 h. Scratch test was used to investigate the effects of 0(blank control ), 10, 20, 30 µmol/L genistein on themigration ability of CNE 1 cells after treated for 24 h. High (No.18210156) throughput sequencing was conducted to discover the differential genes in CNE 1 cells after treated with 0(blankcontrol),30 µmol/L genistein for 24 h. RT-qPCR assay was adopted to verify the mRNA expression of related differential genes in above trials. RESULTS : Compared with blank control,12.5,25,50,100,150 µmol/L genistein sho wed significant inhibitory effect on the proliferation of CNE 1 cells(P< 0.01),in a concentration- time-effect manner ;15,30 µmol/L genistein could arrest CNE 1 cell cycle at G 0/G1 stage(P<0.05 or P< 0.01);30,60 µmol/L could arrest CNE 1 cell cycle at G 2/M stage and promoted cell apoptosis (P<0.05 or P<0.01). 10,20,30 µmol/L genistein could significantly inhibit the migration ability of CNE 1 cells(padj<0.01). High throughput sequencing revealed a total of 2 271 differentialgenes(P<0.05),1 154 of which were up-regulated while 1 117 of which were down-regulated ;8 potential target genes ,including p53,p21,STC2,FGF2,CDK6,CYCLIN D ,PI3K,AKT,were screened by cell experiment. After validated by RT-qPCR assay ,mRNA expression of p53,p21,STC2,FGF2,CDK6,CYCLIN D and AKT were significantly down-regulated(P<0.05),which consistent with the sequencing results. CONCLUSIONS :Genistein can effectively inhibit the growth of human nasopharyngeal carcinoma CNE 1 cells,the mechanism of which may associated with inhibiting the expression of mutant gene p53,restoring the function of wild-type P 53 protein and inhibiting the activity of PI 3K/Akt pathway.
9.Feasibility study of ultrasound-guided percutaneous endomyocardial septal cryoablation of isolated porcine hearts
Xiaonan LU ; Jin HE ; Jia ZHAO ; Jing LI ; Lei ZUO ; Mengyao ZHOU ; Rui HU ; Xiaoli ZHU ; Liwen LIU
Chinese Journal of Ultrasonography 2021;30(5):436-440
Objective:To investigate the feasibility of ultrasound-guided percutaneous endomyocardial septal cryoablation of in vitro porcine hearts and to compare its effect with the percutaneous endomyocardial radiofrequency ablation.Methods:Experiment 1: Six in vitro porcine hearts were divided into 1 min ( n=2), 3 min ( n=2) and 5 min ( n=2) groups according to the cryoablation time, and all were subjected to ultrasound-guided percutaneous intra-myocardial septal cryoablation at 100% power respectively. After cryoablation, ultrasound images, the size of the solid dissection of the ice ball, and the size of the necrotic area after melting of the frozen ice ball were measured. Experiment 2: The in vitro porcine hearts were divided into cryoablation group ( n=3) and radiofrequency ablation group ( n=3), and ultrasound-guided percutaneous endomyocardial septal cryoablation and radiofrequency ablation were performed with 100% cryo power and 40 W radiofrequency power, and the extent of complete necrotic area and incomplete necrotic area were compared between the two ablation methods after 1 min. Results:Experiment 1: In the 1 min cryoablation time group ( n=2), the short diameter of the puck measured by ultrasound was (8.00±0.84)mm, the short diameter of the puck measured by solid was (8.38±1.19)mm, and the short diameter of the necrotic zone measured by solid was (8.35±0.83)mm; in the 3 min group ( n=2), the short diameter of the puck measured by ultrasound was (19.4±0.28)mm, and the short diameter of the puck measured by solid was (19.03±0.33)mm, solid measurement of the short diameter of the necrotic zone was (19.16±0.25)mm; in the 5 min group ( n=2), the short diameter of the puck measured under ultrasound was (26.4±2.54)mm, solid measurement of the short diameter of the puck was (26.01±0.24)mm, and solid measurement of the short diameter of the necrotic zone was (24.82±0.25)mm. Randomized blocks analysis of variance was performed on this data and the difference of block Factor b (freezing time: 1 min, 3 min, 5 min) among the three groups was statistically significant( F=505.884, P<0.001). The SNK- q test showed that all three groups differed from each other(all P<0.05). The analysis results for the treatment factors K (measurement modality-ultrasound image measurements, solid anatomical measurements of the puck, and measurements of the necrotic area after melting of the frozen puck) was not statistically significant ( F=0.470, P=0.635). Experiment 2: In the RF ablation group ( n=3), the ratio of incomplete necrotic zone to the radius of the RF ablation area was 0.64±0.01; in the cryoablation group ( n=3), the ratio of incomplete necrotic zone to the radius of the ablation area was 0.26±0.02. The difference was statistically significant( P=0.002) and it can be considered that the incomplete necrotic zone of cryoablation was smaller than that of RF ablation. Conclusions:Percutameous intramyocardial septal cryoablation is controllable in scope, ultrasound image evaluation of ablation area is more accurate and incomplete necrosis area is small, which may have potential applications in cardiac ablation.
10.Evaluation of myocardial microcirculation perfusion in patients with hypertrophic obstructive cardiomyopathy after Liwen procedure by myocardial contrast echocardiography
Lu YAO ; Lei ZUO ; Jing WANG ; Xiaonan LU ; Jia ZHAO ; Jin HE ; Changhui LEI ; Shengjun TA ; Liwen LIU
Chinese Journal of Ultrasonography 2021;30(11):927-931
Objective:To quantitatively evaluate myocardial microcirculation perfusion changes after percutaneous intracardiac septal radiofrequency ablation (PIMSRA, Liwen procedure) in patients with obstructive hypertrophic cardiomyopathy (HOCM) using myocardial contrast echocardiography (MCE) and to further establish the effect of the procedure and predict the long-term prognosis of patients.Methods:A total of 45 patients with HOCM treated by the Liwen procedure in the First Affiliated Hospital of Air Force Medical University (Xijing Hospital) from July 2019 to June 2020 were included. MCE was performed before and 6 months after surgery, respectively. Time-intensity perfusion curve analysis was performed using QLab 10.8 offline software to obtain quantitative parameters of myocardial microcirculation perfusion including myocardial blood volume (A value), myocardial blood flow velocity (β value), and myocardial blood flow (A×β value), then the changes in parameters before and after the procedure were compared.Results:After the Liwen procedure, the mean septal thickness and mean left ventricular free wall thickness were significantly reduced, accompanied by a significant reduction in the left ventricular outflow tract pressure gradient (LVOT-PG) and mitral regurgitation length, and a significant improvement in mitral systolic antegrade motion (SAM) (all P<0.001). In addition, the left ventricular ejection fraction (LVEF) did not deteriorate significantly ( P=0.560) and the E/e′ ratio decreased after the procedure ( P=0.015). Besides, the A values of both the ventricular septum and the left ventricular free wall were not significantly changed compared to those before procedure ( P>0.05), whereas the β values and A×β values were obviously increased ( P<0.05). Conclusions:Myocardial blood velocity and myocardial blood flow in the septum and left ventricular free wall were significantly increased in patients with HOCM after the Liwen procedure, suggesting a significant improvement in myocardial microcirculation perfusion. MCE provides a non-invasive quantitative evaluation parameters of myocardial microcirculation perfusion for the Liwen procedure for the treatment of HOCM.

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