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.Academician WANG Yongyan′s experience in differentiating and treating motor neuron disease from the perspective of
Liwei LIU ; Yuxuan HUANG ; Yipin FAN
Journal of Beijing University of Traditional Chinese Medicine 2025;48(4):552-558
This article summarizes Academician WANG Yongyan′s experience in the differentiation and treatment of motor neuron disease, which can be categorized into flaccidity syndrome, convulsive syndrome, and fei syndrome according to the clinical manifestations. These three syndromes may coexist, and the condition progressively worsens over time, which is believed to be caused by external pathogenic qi, based on "deficient-qi induced stagnation" , and with "toxins damaging collaterals" as the core etiology and pathogenesis. "Toxins damaging collaterals" involves three levels of qi collaterals, blood collaterals, and fluid collaterals, gradually overlapping and affecting the marrow collaterals. Academician WANG Yongyan′s theory is based on syndrome differentiation, breaking down the boundaries of flaccidity, convulsive, and fei syndromes according to different manifestations of the disease, and using the concept of "combined treatment" for treatment. The clinical presentation of motor neuron disease shows a bottom-up trend in the development of the sanjiao, and the combination of visceral syndrome differentiation and sanjiao syndrome differentiation can grasp the progress of the disease comprehensively. During the process of syndrome differentiation, the focus is on the use of xiang thinking, emphasizing the holistic correlation between diseases and syndromes and the integrated effect of reductionist analysis. Treatment is based on xiang differentiation and individualized treatment. The mid-stage of motor neuron disease is the key time point for the treatment of this disease. Based on the clinical symptoms of flaccidity, convulsive, and fei syndromes, where treatment should focus on reinforcing the spleen and kidney, combining moxibustion with herbal medicine. While targeting the disease, treatment should comprehensively apply the methods of "promoting, supplementing, softening, and warming" to eliminate toxins and unblock collaterals, and restore the neural regulation of the brain and spinal cord.
3.Application value of intrathecal synthetic related markers in cognitive dysfunction and various dementia diseases
Yanan LIU ; Wencan JIANG ; Chenxu WANG ; Chunqing SHAO ; Menglue ZHANG ; Wenya JIA ; Yuxuan HUANG ; Jingchun ZHAI ; Jiayi LIAO ; Guojun ZHANG
International Journal of Laboratory Medicine 2024;45(17):2076-2080
Objective To evaluate the value of intrathecal synthetic related markers in patients with mild cognitive impairment(MCI),Alzheimer's disease(AD),and other types of dementia.Methods Retrospec-tively collect the clinical data of 577 patients diagnosed with MCI(MCI group,178 cases),AD(AD group,131 cases),and other types of dementia(other types group,268 cases)from June 2019 to July 2023 in Beijing Tiantan Hospital,Capital Medical University.Oligoclonal zone electrophoresis(OCB)and 24 h intrathecal pro-tein synthesis rate(ISR)of each group were investigated,and the difference of different indexes among the groups was compared to evaluate the value of related indexes in the differential diagnosis of different diseases.Results Compared with AD group and other groups,AD group had a higher proportion of females,more patients were>50-70 years old,and the incidence of abnormal lipid metabolism was higher,with statistical significance(P<0.05).There were significant differences in OB(S),cerebrospinal fluid albumin,serum albumin and cerebrospinal flu-id IgG in different disease groups(P<0.05).IgG index and ISR in patients with positive SOB(CSF)were higher than those in negative and weakly positive patients,and the differences were statistically significant(P<0.001).IgG index was positively correlated with ISR(r=0.878,P<0.001).Conclusion Intrathecal synthetic mark-ers such as IgG index,SOB(CSF)and 24 h ISR have synergistic effects in the diagnosis of cognitive dysfunction and various dementias,which can be collectively utilized in the diagnosis of diseases.
4.Aerobic exercise modulates changes in cardiac function due to simulated weightlessness through the AMPK/ULK1/Beclin1 pathway
Yequn LIU ; Jianjun YANG ; Min YANG ; Hongli LI ; Fudong GAO ; Yuxuan HUANG ; Shuling ZHANG ; Junzhi SUN
Space Medicine & Medical Engineering 2024;35(4):235-240
Objective To explore the possible mechanisms of myocardial injury induced by simulated weightlessness in space flight and the recovery of myocardium in rats by aerobic exercise intervention.Methods The body weight of rats was weighed using an electronic balance;the cardiac function indexes of rats in each group were detected using echocardiography and isolated cardiac perfusion;and the expression of proteins related to the AMPK/ULK1/Beclin1 pathway in the myocardial tissues of rats in each group was detected using Western Blot.Results(1)When compared with the C1 group,the T1 group not statistically significant in body weight(P?>?0.05),and the wet weight of the flounder muscle,the wet weight-to-weight ratio of the flounder muscle,and the heart weight were all significantly decreased(P?0.01).(2)After 4 weeks of tail suspension and 4 weeks of aerobic exercise,significant changes in cardiac function-related indexes occurred,with a significant increase in the expression levels of AMPK and p-ULK1 in the T1 group(P?0.01),a significant increase in the expression of p-AMPK(P?0.05),and a slight,but not statistically significant,increase in the expression of ULK1 and Beclin1(P?>?0.05).After 4 weeks of aerobic exercise,when compared to group C2,group R showed a significant rise in the expression of AMPK and ULK1(P?0.05),a decrease in the expression of p-AMPK and p-ULK1,but not statistically significant(P?>?0.05),and a rise in the expression of Beclin1,but not statistically significant(P?>?0.05);and group A showed a slight increase in the expression of AMPK,ULK1 and Beclin1 expression decreased slightly but not statistically significant(P?>?0.05),p-AMPK expression decreased significantly(P?0.05)and p-ULK1 expression decreased significantly(P?0.01)in Group A.When compared with group R,group C2 showed a significant decrease in the expression of cardiac protein AMPK(P?0.05),a slight increase in the expression of p-AMPK and p-ULK1,a statistically insignificant decrease in the expression of Beclin1(P?>?0.05),and a significant decrease in the expression of ULK1(P?0.01);and group A showed a significant decrease in the expression of AMPK,ULK1(P?0.01);the expression of p-AMPK and Beclin1 decreased slightly but not statistically significant(P?>?0.05);the expression of p-ULK1 decreased significantly(P?0.05).Conclusion(1)After 4 weeks of tail suspension,it was found that the wet weight,and wet weight to body weight ratio of the flounder muscle of rats were significantly reduced,the weight of the heart and cardiac function related indexes were also significantly reduced,and the expression of AMPK,p-AMPK,ULK1,p-ULK1 and Beclin1 increased;(2)After 4 weeks of aerobic exercise,it was found that the morphology of the myocardium of the rat with tail suspension,the myocardial ultrastructure and cardiac function related indexes increased,and cardiac function was improved.function-related indexes rose,cardiac function was somewhat improved,and the expression of AMPK,p-AMPK,ULK1,p-ULK1,and Beclin1 declined;(3)he damage and improvement of cardiac function were associated with the regulation of related proteins through the AMPK/ULK1/Beclin1 pathway,and the regulation of the expression of AMPK,p-AMPK,ULK1,p-ULK1,and Beclin1.
5.Computer-aided design of an improved lamina hook and finite element analysis of its use in fixation of lumbar spondylolysis
Hongliang GAO ; Hua LIU ; Tao ZHANG ; Chengwei YANG ; Yizhe WANG ; Zirong HUANG ; Wenhua ZHANG ; Long CHEN ; Bing KANG ; Yuxuan MA ; Songkai LI
Chinese Journal of Trauma 2024;40(7):593-604
Objective:To design an improved lamina hook system and compare its biomechanical properties with traditional lamina hook system in fixation of lumbar spondylolysis.Methods:The thin layer CT data of the lumbosacral vertebrae of 20 healthy young male servicemen who underwent physical examination in the outpatient department of the 940th Hospital of Joint Logistics Support Force of PLA from January 2021 to August 2022 were collected. The age of the subjects was 20-30 years [(25.0±3.0)years]. A 3-dimensional model of the L 5 vertebral body was constructed using the 3-dimensional modeling software. The new improved lamina hook was designed according to the measurements including the thickness of the middle area, the longest longitudinal diameter, the curvature radius of the lower edge, the angle between the upper and lower tail ends, the thickness of the lower edge, and the longest diameter of the lower edge of the bilateral L 5 vertebral plates. One serviceman was selected from the aforementioned group to construct a linear finite element model of segments L 4-S using the 3-dimensional virtual software (normal model, model A), based on which, the L 5 bilateral spondylolysis model (model B), improved lamina hook model (model C) and traditional lamina hook models (model D) were designed. By constraining both sides of the sacrum and applying a longitudinal load of 400 N on the L 4 vertebral body, the upper 1/3 gravity of the body was simulated, and with a bending moment of 10 N·m along the X, Y, and Z directions, motions of forward flexion, backward extension, lateral bending, rotation, etc were simulated. The range of motion of segment L 4/5 and L 5/S 1 of model A was evaluated and compared with the findings of the previous researches to verify its effectiveness. The overall range of motion of models A, B, C, and D, the range of motion of segment L 4/5 and L 5/S 1, the maximum overall displacement, the maximum displacement and stress of the isthmus, the stress distribution and maximum stress of internal fixation of models C and D, and the stress distribution and maximum stress of the vertebral body of models C and D were compared. Results:(1) During forward flexion, backward extension, lateral bending and rotation, the range of motion of model A was 5.01°, 4.03°, 3.91° and 1.42° in segment L 4/5, and was 4.62°, 2.51°, 2.40° and 1.23° in segment L 5/S 1. (2) The overall range of motion, range of motion of segment L 4/5 and L 5/S 1 and maximum overall displacement of models A, C, and D were similar in axial compression, forward flexion, backward extension, left bending, and left rotation, while those of model B were significantly increased. (3) There was no significant difference in the maximum displacement of the isthmus of models A, C, and D under different motion modes, while the maximum displacement of model B in the isthmus was significantly larger than that of models A, C, and D, especially during rotation, increased by 295%, 277%, and 276% respectively. The maximum stress of the isthmus of model C was 0.938 MPa, 1.698 MPa, 0.410 MPa, 2.775 MPa, and 1.554 MPa respectively. The maximum stress in the isthmus of model D was 0.590 MPa, 1.297 MPa, 0.520 MPa, 3.088 MPa, and 2.072 MPa respectively. The maximum stress of the isthmus of models C and D was similar during axial compression and forward flexion, while the stress of the isthmus of model C was smaller than that of model D during backward extension, lateral bending, and rotation, decreased by 21.1%, 10.2%, and 25.0% respectively compared with model D. (4) The maximum stress of internal fixation in models C and D during forward flexion, backward extension, left bending, and left rotation was 135.220 MPa, 130.180 MPa, 200.940 MPa and 306.340 MPa respectively, and was 131.840 MPa, 112.280 MPa, 349.980 MPa and 370.140 MPa respectively. The maximum stress of internal fixation in the two models of internal fixation during forward flexion and backward extension was similar, while it was decreased by 42.6% and 17.2% in model C during left bending and left rotation, compared with model D. (5) The maximum stress of the vertebral body during forward flexion, backward extension, left bending, and left rotation was 79.787 MPa, 36.857 MPa, 37.943 MPa and 96.965 MPa respectively in model C, but was 80.104 MPa, 64.236 MPa, 196.010 MPa and 193.020 MPa respectively in model D. The maximum stress of models C and D was all distributed in the contact area with the internal fixation, and especially during backward extension, left bending, and left rotation, when it was reduced by 42.6%, 80.6%, and 49.8% of model C respectively, compared with that of model D. Conclusions:The improved laminar hook is more consistent with the Chinese anatomized structure of the lamina. Compared with the traditional lamina hook system, the improved lamina hook system can effectively reduce the displacement in all directions and range of motion of lumbar spondylolysis, therefor can significantly reduce the stress of internal fixation and vertebral body and has better biomechanical performance.
6.Progress in the study of animal models of dwarfism
Yifan LIU ; Nan HUANG ; Yuxuan LIU ; Ling GAN ; Hongyu CHEN
Acta Laboratorium Animalis Scientia Sinica 2024;32(9):1207-1214
Dwarfism is a globally rare growth disorder,usually caused by genetics or disease,with the most prominent phenotype being short stature.Animal models are important tools for studying its pathogenesis,prevention,and treatment options,and identifying potential therapeutic targets and biomarkers.The development of genetic engineering technology has greatly promoted the application of gene-edited animal models in the study of dwarfism.In this review,we summarize and discuss the existing animal models of dwarfism in terms of their theoretical basis,model characteristics,and research applications.This offers a reference for researchers and clinicians aiming to better conduct research on the pathogenesis and prevention of dwarfism.
7.Fibrinolytic effect of curdione in dissolving deep vein thrombosis in rats
Hanbing SHI ; Teng ZHANG ; Shan DU ; Yuxuan GUAN ; Yan HUANG ; Quan XIA
China Pharmacist 2024;28(11):424-430
Objective To investigate whether curdione inhibits the formation of deep vein thrombosis(DVT)in rats through thrombolytic effects.Methods Twenty-five rats were randomly divided into five groups,the normal control group,the sham operation group,the model group,the low dose group of curdione(60 mg/kg)and the high dose group of curdione(120 mg/kg),with five rats in each group.Using the inferior vena cava ligation model,blood was taken from the pulmonary aorta in a sodium citrate anticoagulant tube one week after drug administration.The coagulation and fibrinolytic indexes Fibrin Degradation Products,(FDP),D-dimer(D-D),plasminogen activator inhibitor(PAI)-1 and plasminogen(PLG)were detected in each group;the thrombus weight/weight-length ratio was detected;and the formation of venous thrombus in the lower limbs of rats was detected by hematoxylin-eosin(HE)staining nuclear factor;NF-κB protein expression in inferior vena cava tissues was dectected by Western Blot assay.Results There was no significant difference in plasma FDP levels among groups of rats(P>0.05);compared with the model group,the plasma FDP and D-D levels of rats in each dosing group were increased,the plasma PAI-1 and PLG levels were elevated,and the fibrinolytic effect was better in the group with high dose of curdione administration than in the group with low dose of curdione administration(P<0.05);compared with the model group,the thrombus weight/weight-length ratio,the thrombus mass and NF-κB protein expression in the curdione dosing group were decreased.Conclusion Curdione can inhibit the formation of DVT in rats by influencing the fibrinolytic regulators.
8.Application of Modified FuMai Decoction (复脉汤加减方) based on Sanjiao (三焦) Theory
Jienan GU ; Yuxuan HE ; Bin PENG ; Sheng HUANG ; Guowei WANG ; Yongtao WANG ; Qianhui LIU ; Shijie XU
Journal of Traditional Chinese Medicine 2023;64(20):2157-2160
Review of historical literature showed that the understanding of the indicated disease location of Modified FuMai Decoction (复脉汤加减方) has evolved from the upper jiao (焦) to the middle and lower jiao. Initially, it is used for the treatment of yin deficiency of both yin and yang in the upper jiao, changes to supplement stomach and produce fluids in the middle jiao, and is used to protect yin, clear the pathogens, conslidate yin and subdue yang so as to store the true yin of lower jiao. The unchanging principle of Fumai Decoction modifications is nourishing yin, while the changing aspects are determining the secondary treatment methods based on disease location of sanjiao, concomitant disease natures, internal injury or external contraction, warm disease or cold damage, thereby choosing the corresponding added or subtracted herbs, and providing reference for the application of classical formulas.
9.A multicenter study to test the reliability and validity of the frailty assessment scale for elderly patients with inguinal hernia and to evaluate the value of clinical application
Xian LI ; Jia ZHENG ; Shibo WEI ; Hangyu LI ; Lei JIANG ; Lei DONG ; Jiang WANG ; Chongzhu TAO ; Yuhao YAN ; Lihui SUN ; Lunbo CUI ; Jinhai HUANG ; Yuxuan FANG ; Chengxin TANG
Chinese Journal of Surgery 2023;61(12):1080-1085
Objectives:To verify the reliability and validity of the frailty assessment scale for elderly patients with inguinal hernia and to evaluate the value of its clinical application.Methods:A convenience sampling method was used to collect 129 geriatric patients who underwent inguinal hernia surgery from January 2018 to January 2023 in nine hospitals in Liaoning Province. There were 120 males and 9 females, of whom 89 patients were 60 to <75 years old, 33 patients were 75 to <85 years old and 7 patients were ≥85 years old. The 129 patients included 11 elderly patients with inguinal hernia who had recovered from preoperative infection with COVID-19. Statistical methods such as Cronbach′s coefficient, Kaiser-Meyer-Olkin test, Bartlett′s test, Pearson′s correlation analysis, etc. were calculated to verify the reliability indexes such as feasibility, content validity, structural validity, criterion-related validity, internal consistency reliability, and re-test reliability. Taking the 5-item modified frailty index (5-mFI) as the gold standard, the area under the curve was used to analyze the ability of the two scales to predict the occurrence of postoperative acute urinary retention, postoperative delirium, poor incision healing, operative hematoma seroma, and postoperative complications.Results:The frailty assessment scale for elderly patients with inguinal hernia showed good reliability and validity (valid completion rate of 99.2%; item content validity index of 1.000, and the scale content validity index of 1.000; exploratory factor analysis extracted a total of 1 principal component, and factor loadings of each item of 0.565 to 0.873; the AUC for frailty diagnosis using 5-mFI as the gold standard of 0.795 ( P<0.01) Cronbach′s coefficient of 0.916, retest reliability coefficient of 0.926), it could effectively predict postoperative acute urinary retention, delirium, hematoma seroma in the operative area and total complications (AUC of 0.746, 0.870, 0.806, and 0.738, respectively; all P<0.05), and prediction efficiency was higher than that of 5-mFI (AUC of 0.694, 0.838, 0.626 and 0.641, P<0.05 for delirium only), but both scales were inaccurate in predicting poor incision healing (AUC of 0.519, P=0.913 for the frailty assessment scale and 0.455, P=0.791 for the 5-mFI). Conclusions:The frailty assessment scale for elderly patients with inguinal hernia is reliable and significantly predicts the occurrence of postoperative adverse events in elderly inguinal hernia patients. The scale can also be used for preoperative frailty assessment in elderly patients with inguinal hernia after rehabilitation from COVID-19 infection.
10.A multicenter study to test the reliability and validity of the frailty assessment scale for elderly patients with inguinal hernia and to evaluate the value of clinical application
Xian LI ; Jia ZHENG ; Shibo WEI ; Hangyu LI ; Lei JIANG ; Lei DONG ; Jiang WANG ; Chongzhu TAO ; Yuhao YAN ; Lihui SUN ; Lunbo CUI ; Jinhai HUANG ; Yuxuan FANG ; Chengxin TANG
Chinese Journal of Surgery 2023;61(12):1080-1085
Objectives:To verify the reliability and validity of the frailty assessment scale for elderly patients with inguinal hernia and to evaluate the value of its clinical application.Methods:A convenience sampling method was used to collect 129 geriatric patients who underwent inguinal hernia surgery from January 2018 to January 2023 in nine hospitals in Liaoning Province. There were 120 males and 9 females, of whom 89 patients were 60 to <75 years old, 33 patients were 75 to <85 years old and 7 patients were ≥85 years old. The 129 patients included 11 elderly patients with inguinal hernia who had recovered from preoperative infection with COVID-19. Statistical methods such as Cronbach′s coefficient, Kaiser-Meyer-Olkin test, Bartlett′s test, Pearson′s correlation analysis, etc. were calculated to verify the reliability indexes such as feasibility, content validity, structural validity, criterion-related validity, internal consistency reliability, and re-test reliability. Taking the 5-item modified frailty index (5-mFI) as the gold standard, the area under the curve was used to analyze the ability of the two scales to predict the occurrence of postoperative acute urinary retention, postoperative delirium, poor incision healing, operative hematoma seroma, and postoperative complications.Results:The frailty assessment scale for elderly patients with inguinal hernia showed good reliability and validity (valid completion rate of 99.2%; item content validity index of 1.000, and the scale content validity index of 1.000; exploratory factor analysis extracted a total of 1 principal component, and factor loadings of each item of 0.565 to 0.873; the AUC for frailty diagnosis using 5-mFI as the gold standard of 0.795 ( P<0.01) Cronbach′s coefficient of 0.916, retest reliability coefficient of 0.926), it could effectively predict postoperative acute urinary retention, delirium, hematoma seroma in the operative area and total complications (AUC of 0.746, 0.870, 0.806, and 0.738, respectively; all P<0.05), and prediction efficiency was higher than that of 5-mFI (AUC of 0.694, 0.838, 0.626 and 0.641, P<0.05 for delirium only), but both scales were inaccurate in predicting poor incision healing (AUC of 0.519, P=0.913 for the frailty assessment scale and 0.455, P=0.791 for the 5-mFI). Conclusions:The frailty assessment scale for elderly patients with inguinal hernia is reliable and significantly predicts the occurrence of postoperative adverse events in elderly inguinal hernia patients. The scale can also be used for preoperative frailty assessment in elderly patients with inguinal hernia after rehabilitation from COVID-19 infection.


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