1.Effect of lentiviral silencing of Piezo1 on osteogenic differentiation and TAZ expression in human bone marrow mesenchymal stem cells
Yurou WEI ; Jiaqing TIAN ; Xianshun HE ; Zhiwei ZHAN ; Tengfei WEI ; Tianye LIN ; Wei HE ; Qiushi WEI
Chinese Journal of Tissue Engineering Research 2024;28(1):12-19
BACKGROUND:Piezo1,a mechanosensitive protein,is tightly connected to osteogenic differentiation,and it has been demonstrated that TAZ has a role in regulating osteogenic differentiation.It is unclear whether TAZ participates in the regulation of osteogenic differentiation of human bone marrow mesenchymal stem cells by Piezo1,so it is crucial to investigate its unique mechanism to prevent osteonecrosis of the femoral head. OBJECTIVE:To elucidate what function Piezo1 plays in osteogenic differentiation and TAZ expression in human bone marrow mesenchymal stem cells. METHODS:The siRNA targeting Piezo1 was constructed and transfected into 293T cells.The silencing efficiency was detected by RT-qPCR.The selected Piezo1-Home-2337 was packaged according to the silencing efficiency,and its optimal multiplicity of infection value was assayed by immunofluorescence staining.The packaged Piezo1 silencing recombinant lentivirus was transfected into human bone marrow mesenchymal stem cells,and its silencing effect was detected by RT-qPCR and western blot assay.Alizarin red staining,alkaline phosphatase activity analysis,immunofluorescence staining,RT-qPCR and western blot assay were utilized to analyze the effect of silencing Piezo1 on the osteogenic differentiation of human bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION:(1)The mRNA and protein levels of Piezo1 in human bone marrow mesenchymal stem cells transfected by si-Piezo1 were decreased significantly,with a statistically significant difference compared with normal and negative control groups.(2)The alkaline phosphatase activity in the si-Piezo1 group was much lower and the calcium deposition in the si-Piezo1 group was significantly reduced compared with the negative control group.(3)The mRNA levels of osteogenesis-related genes including Runt-related transcription factor 2(Runx2),osteopontin(OPN),distal-less homeobox 5(DLX5),osteocalcin,β-catenin and Tafazzin(TAZ)in the si-Piezo1 group were significantly decreased compared with the negative control group.Afterward,the expression levels of TAZ and β-catenin protein in the si-Piezo1 group were down-regulated significantly compared with the negative control group,whereas the expression levels of p-TAZ and p-β-catenin protein in the si-Piezo1 group had the opposite condition.(4)The results of immunofluorescence staining showed that the expression of TAZ and β-catenin in human bone marrow mesenchymal stem cells in the si-Piezo1 group was less compared with the negative control group.(5)These findings indicate that Piezo1 can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells.The osteogenic ability of human bone marrow mesenchymal stem cells is significantly reduced after silencing Piezo1,and the expression of TAZ is also reduced.
2.Progress in animal models of hemophilic arthritis
Kun LIN ; Xianshun HE ; Jiaqing TIAN ; Shun LU ; Mincong HE ; Tianye LIN ; Xiaoming HE ; Shuai FAN ; Hongguang YANG ; Tianqi YU ; Daguang ZHANG ; Wei HE ; Qiushi WEI
Acta Laboratorium Animalis Scientia Sinica 2024;32(4):523-529
Hemophilic arthritis(HA),caused by recurrent bleeding,can seriously affect patient quality of life and consumes extensive social and medical resources.There is thus a need to establish an animal model of HA for research;however,this is limited by ethical requirements.Here we review the recent literature and summarize research progress into animal models of HA at home and abroad,from the aspects of species selection,modeling method,histopathology,and imaging evaluation method.Species selection includes rodents such as mice,New Zealand rabbits,beagles,miniature pigs,and crab-eating macaques.Modeling method comprise gene knockout trauma models,gene knockout spontaneous models,and injection models.Among these,the gene knockout spontaneous model closely mimics the pathological process of spontaneous bleeding and concurrent arthritis in human HA,making it more relevant to human HA.However,due to high modeling costs,phenotypic instability,and low survival rates,this model is not the preferred choice for animal experimental studies.In contrast,gene knockout trauma models exhibit characteristics such as short modeling time,strong stability,and high success rates,thus being widely utilized in animal experimental research.Evaluation of HA models involves various imaging method including MRI,micro-CT,MSKUS/PD,in addition to various gross scoring method.By reviewing the progress of HA model research,more experimental evidence is provided for investigating the pathogenesis and validating the efficacy of HA treatments,thereby compensating for the lack of clinical data,particularly in the field of traditional Chinese medicine therapy.
3.Therapeutic effects of Qingre jiedu formula on mice with gout based on immune inflammation and gut microbiota
Xianshun HE ; Kun LIN ; Shun LU ; Yurou WEI ; Jiaqing TIAN ; Yulai JIANG ; Tengfei WEI ; Tianye LIN ; Mincong HE ; Qiushi WEI
Chinese Journal of Comparative Medicine 2024;34(5):1-12
Objective To explore the anti-inflammatory effect of Qingre Jiedu(QRJD)formula on mice with gout and its effect on gut microbiota.Methods Forty C57BL/6 mice weighing 20~22 g were divided into control(CON),model(MOD),allopurinol(ALLO),and QRJD formula(QRJD)groups,and i.g.10 g/0.1 mL carboxymethyl cellulose was administered to the CON every morning from 1 to 35 days.A hyperuricemia mouse model was prepared by intragastric injection of a potassium oxalic acid(500 mg/kg)and yeast extract(10 g/kg)suspension.On day 29,50 μL sterile carboxymethyl cellulose was injected into the right ankle of mice in the CON group under isoflurane-induced anesthesia,and a gouty arthritis model was prepared by injecting the same volume of sodium urate(50 mg/mL)into the right ankle of mice in the other groups.Each group was treated with corresponding drugs every day.On day 35,samples were collected from mice that had been fasted for 6 hours without water.Blood indexes,such as uric acid,creatinine,and urea nitrogen,were assessed.Hematoxylin-eosin and saffranine O-fast green staining was performed on ankle joints.Anti-inflammatory indexes of interleukin-10(IL-10)and transforming growth factor-β1(TGF-β1)were detected by ankle joints immunohistochemical assay.The cecum contents of mice were collected,and changes in gut microbiota were analyzed by high-throughput sequencing of 16S rDNA.Results(1)After 7 days of treatment,compared with the MOD group,QRJD formula effectively reduced the blood concentrations of uric acid(P<0.001),creatinine(P<0.01),and urea nitrogen(P<0.05),and effectively protected renal functions.(2)Compared with the MOD group,HE staining showed that synovial hyperplasia and inflammatory cell infiltration were reduced in the QRJD formula group after treatment.The cartilage arrangement of the compound was more orderly than before,cartilage destruction was less than that in the MOD group,and no matrix loss was observed.(3)Immunohistochemical analysis of the ankle joint indicated that IL-10 and TGF-β1 were not significantly increased in CON and MOD groups.Compared with the MOD group,IL-10 and TGF-β1 expression in the QRJD formula group were increased(P<0.05).(4)In terms of biodiversity,the number of MOD-specific OTUs increased by 75 compared to the CON group,while the QRJD was able to reduce the number of MOD-specific OTUs to more closely resemble the CON group;no significant difference was found in α-diversity among the four groups(P<0.05),whereas β-diversity was more similar to the CON group(P=0.001).(5)Compared with the CON group,the MOD group exhibited increased abundances(P<0.05)of Ruminococcaceae spp.,Dubosiella sp.,Tyzzerella sp.,Ileibacterium sp.,and Bacteroidales spp..In contrast to the MOD group,the QRJD formula group showed elevated abundances(P<0.05)of Lactobacillus sp.,Ligilactobacillus sp.,and Bacteroides sp..Furthermore,an interaction network of gut microbiota indicated mutual interactions among these microorganisms.(6)In the correlation analysis between gut microbiota and renal functions as well as anti-inflammatory factors,the relative abundances of Dubosiella sp.,Tyzzerella sp.,and Bacteroidales spp.were significantly positively correlated to SUA and SCR(P<0.05).However,Lactobacillus sp.,Ligilactobacillus sp.,and Mitochondria spp.exhibited a positive correlation to anti-inflammatory factors IL-10 and TGF-β1 with a more significant association observed for TGF-β1(P<0.05).(7)COG function prediction suggested that the functions of the QRJD formula group were concentrated on inorganic ion transport and metabolism,and carbohydrate transport and metabolism.Conclusions QRJD effectively modulates immune inflammation and gut microbiota dysbiosis,thereby treating gout.Its mechanism of gout prevention and treatment may involve regulation of gut microbiota diversity and abundance,as well as the control of the abundance of differential bacterial species,such as Ruminococcaceae spp.,Dubosiella sp.,and Lactobacillus sp.,to achieve gout therapy.
4.Finite element analysis for predicting osteonecrosis of the femoral head collapse based on the preserved angles.
Shun LU ; Tianye LIN ; Mincong HE ; Xiaoming HE ; Xianshun HE ; Jiaqing TIAN ; Tengfei WEI ; Zhiwei ZHAN ; Kun LIN ; Qiushi WEI
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(11):1394-1402
OBJECTIVE:
To establish finite element models of different preserved angles of osteonecrosis of the femoral head (ONFH) for the biomechanical analysis, and to provide mechanical evidence for predicting the risk of ONFH collapse with anterior preserved angle (APA) and lateral preserved angle (LPA).
METHODS:
A healthy adult was selected as the study object, and the CT data of the left femoral head was acquired and imported into Mimics 21.0 software to reconstruct a complete proximal femur model and construct 3 models of necrotic area with equal volume and different morphology, all models were imported into Solidworks 2022 software to construct 21 finite element models of ONFH with LPA of 45°, 50°, 55°, 60°, 65°, 70°, and 75° when APA was 45°, respectively, and 21 finite element models of ONFH with APA of 45°, 50°, 55°, 60°, 65°, 70°, 75° when LPA was 45°, respectively. According to the physiological load condition of the femoral head, the distal femur was completely fixed, and a force with an angle of 25°, downward direction, and a magnitude of 3.5 times the subject's body mass was applied to the weight-bearing area of the femoral head surface. The maximum Von Mises stress of the surface of the femoral head and the necrotic area and the maximum displacement of the weight-bearing area of the femoral head were calculated and observed by Abaqus 2021 software.
RESULTS:
The finite element models of ONFH were basically consistent with biomechanics of ONFH. Under the same loading condition, there was stress concentration around the necrotic area in the 42 ONFH models with different preserved angles composed of 3 necrotic areas with equal volume and different morphology. When APA was 60°, the maximum Von Mises stress of the surface of the femoral head and the necrotic area and the maximum displacement of the weight-bearing area of the femoral head of the ONFH models with LPA<60° were significantly higher than those of the models with LPA≥60° ( P<0.05); there was no significant difference in each index among the ONFH models with LPA≥60° ( P>0.05). When LPA was 60°, each index of the ONFH models with APA<60° were significantly higher than those of the models with APA≥60° ( P<0.05); there was no significant difference in each index among the ONFH models with APA≥60° ( P>0.05).
CONCLUSION
From the perspective of biomechanics, when a preserved angle of ONFH is less than its critical value, the stress concentration phenomenon in the femoral head is more pronounced, suggesting that the necrotic femoral head may have a higher risk of collapse in this state.
Adult
;
Humans
;
Femur Head/surgery*
;
Finite Element Analysis
;
Stress, Mechanical
;
Femur/diagnostic imaging*
;
Femur Head Necrosis/surgery*
5.Analysis of the status quo of clinical nurses' disaster nursing ability and its influencing factors
Jiaqing XU ; Yingying GAO ; Lingyu DAI ; Chengyuan HE ; Ruixuan XIANG ; Wenjuan LAI
Chinese Journal of Practical Nursing 2021;37(27):2104-2110
Objective:To explore the current situation of clinical nurses' disaster nursing ability and its influencing factors, and to provide basis for carrying out clinical nurses' disaster nursing education and training.Methods:By convenient sampling method, 322 clinical nurses from Peking University Shenzhen Hospital in September 2020 were selected. The disaster nursing ability assessment tool and general information questionnaire were used to conduct a questionnaire survey to analyze the current situation of clinical nurses' disaster nursing ability and its main influencing factors.Results:The total score of disaster nursing ability of clinical nurses was (199.07±34.31) points, which was at the middle and lower level; the scores of each dimension from high to low were as follows: disaster preparedness score was (72.94±12.89) points, coping ability score was (80.00±13.82) points, disaster reduction/prevention capability score was (28.19±6.09) points, recovery/reconstruction ability score was (17.94±3.27) points. Regression analysis showed that the highest degree, clinical department, whether they have autonomously studied disaster nursing related courses after work, and whether they have participated in disaster rescue after work were the main influencing factors of clinical nurses' disaster nursing ability ( t values were -4.715-5.508, P<0.05). Conclusions:The overall level of disaster nursing ability of clinical nurses is at the middle and lower level. It should be combined with clinical nurses' work department, whether clinical nurses have disaster rescue experience and whether they have independently studied disaster nursing related courses, and targeted disaster nursing related training should be carried out to improve their disaster nursing knowledge and skill.
6.Analysis of self-efficacy and influencing factors of male nurses
Jiaqing XU ; Ruixuan XIANG ; Wenting ZHANG ; Chengyuan HE ; Lingyu DAI ; Wenjuan LAI ; Xiaorong DING
Chinese Journal of Practical Nursing 2021;37(32):2488-2493
Objective:To investigate and analyze the current situation and influencing factors of professional decision-making self-efficacy of male nurses.Methods:Convenience sampling was used to select 133 male nurses from September to October 2020 in Shenzhen City as the research objects. The Self-efficacy Scale for Career Decision-making was used to conduct self-evaluation, career information collection, career goal selection, career planning formulation, and job-selection problem resolution.Results:Attitudes towards nursing majors, academic qualifications, reasons for applying for nursing majors, monthly family income, whether it is an only child, family residence, work status, number of job changes, mother′s education level, and married or not were the factors that affect male nurses′ professional self-efficacy ( t values were -1.989-12.523, F values were 7.476-325.316, P<0.05 or 0.01). Conclusion:Career decision-making self-efficacy has a good guiding role in the career selection, development and planning of male nurses. Medical units should formulate reasonable training methods or related training for new male nurses entering the clinic, so as to increase male nurses′ recognition of their occupations, thereby enhancing them Career decision-making self-efficacy.
7.Effects of BCL6 B on proliferation and migration of human colorectal car-cinoma LoVo cells and its potential mechanism
Yue GU ; Aifang LI ; Hui SUN ; Xueru LI ; He ZHA ; Jiali ZHAO ; Jiaqing XIE ; Lan ZHOU
Chinese Journal of Pathophysiology 2017;33(1):38-45
AIM:To detect the endogenous expression of B-cell leukemia/lymphoma 6 member B (BCL6B) in FHC and LoVo cells, and to investigate the effects of BCL6B on proliferation and migration of LoVo cells for further explo-ring the underlying mechanism .METHODS:The endogenous expression of BCL 6B in the FHC and LoVo cells was detec-ted by RT-PCR and Western blot .The methods of MTT assay , colony formation assay , wound healing assay and Transwell chamber experiment were employed to examine the biological functions of BCL 6B in the LoVo cells.The mRNA and protein levels of BCL6B, cyclin D1 and matrix metalloproteinase-9 ( MMP-9) were determined by RT-PCR and Western blot , re-spectively.The level of phosphorylated protein kinase B (p-AKT) was detected by Western blot.RESULTS:BCL6B ex-pression was notably repressed in the LoVo cells as compared with the FHC cells , which were significantly increased by transfection with pcDNA3.1-BCL6B.The abilities of proliferation and migration of the LoVo cells at 72 h were inhibited by 28.33%(P<0.01) and 36.11%(P<0.05) in BCL6B group.The mRNA levels of cyclin D1 and MMP-9 in the cells of BCL6B group were decreased by 39.90%(P<0.01) and 77.36% (P <0.05), and the protein levels of cyclin D1, MMP-9 and p-AKT were reduced by 44.00%(P<0.05), 47.06%(P<0.01) and 32.88% (P<0.05), respectively. CONCLUSION:BCL6B inhibits proliferation and migration of the LoVo cells , and the PI3K/AKT signaling pathway is in-volved in this process .
8.Status and influencing factors of contracting and renewing on family doctor service in pilot areas in Guangzhou
Yujing NIU ; Miaowen YU ; Yue QIU ; Jin CAI ; Hongzheng HE ; Xingjun HUANG ; Zhaofei DING ; Jiaqing MIAO ; Yuanyi LIN ; Li LI
Chinese Journal of Hospital Administration 2017;33(12):939-943
Objective To learn the current status of family doctor service at pilot communities in Guangzhou, and discover existing problems and influencing factors by investigating the residents who have contracted such service , those have not and the family doctors . Methods This study chose typical community health centers of six communities in Guangzhou in January 2016 .In random sampling , residents who visited doctors during the survey and all the family doctors were surveyed .EpiData was used to doubly inputdata,withSPSS20.0forstatisticalanalysis.Results 66.0%ofthoseresidentswhohavenot contracted the service are willing to contract a family doctor .According to the binary logistic regression analysis after eliminating the interference factors , there are two factors affecting their willingness:gender and whether needing a family doctor for themselves and their family for health management .According to the binary logistic regression analysis after eliminating the interference factors , the influencing factors of renewing contract are overall satisfaction and necessity for signing family doctors .Conclusions The smooth development of the family doctor service is faced with many bottlenecks , while improving willingness to contract and renew contract to family doctors are the cornerstone for sustainability of the family doctor system.
9.Effects of effective fraction of Epimedium,Astragalus,Radix Puerariae on the expression of DMT1 in the cerebral cortex of transgenic mice model of Alzheimer′s disease
Xianhui DONG ; Weijuan GAO ; Xiaoping HE ; Zhichao CUI ; Xiqing CHAI ; Jiaqing ZHAO
Chongqing Medicine 2016;45(16):2176-2179
Objective To investigate the effects of Epimedium ,Astragalus ,Radix Puerariae on DMT1 expression in the cere‐bral cortex of APPswe/PS1ΔE9 double transgenic mice model of AD .Methods A total of 60 specific‐pathogen‐free male APPswe/PS1ΔE9 double transgenic mice aged 6 months were equally and randomly assigned to model ,Epimedium ,Astragalus ,Radix puerari‐ae ,compound and DFO groups .An additional 10 6‐month‐old C57BL/6J mice served as negative control group .Using immunohisto‐chemistry and molecular biology methods to investigate the effects of a compound combining the effective components of Epimedi‐um ,Astragalus ,Radix puerariae on DMT1 expression in the cerebral cortex of APPswe/PS1ΔE9 double transgenic mice model of AD . Results Immunohistochemical staining results revealed that DM T 1 positive cell did not show in negative control group .DM T1 ex‐pression was higher in model group compared with the negative control group .DMT1 expression was lower in the compound and deferoxamine groups than in the model group .No significant difference was detected in DM T 1 expression between deferoxamine and compound groups .RT‐PCR ,Western blot and immunohistochemical staining results showed no significant difference .Conclusion These compounds can downregulate DMT1 expression and inhibit iron overload in the cerebral cortex of mice with Alzheimer′s dis‐ease ,reduce iron overload induced impairment of the central nervous system .
10.Clinical significance of soluble B7-H3 in early evaluating the severity of patients with acute pancreatitis
Jian HUANG ; Guangbo ZHANG ; Guangsheng HE ; Jiaqing SHEN ; Xiaoming KONG ; Yueping SHEN
The Journal of Practical Medicine 2014;(22):3569-3572
Objective To investigate the clinical value of sB7-H3 in predicting the severity of acute pancreatitis at early stage. Methods By using the double antibody sandwich ELISA method, the level of plasma sB7-H3 was measured at 24h after onset of abdominal pain in 75 patients with acute pancreatitis (MAP30, MSAP20, SAP25), and 20 healthy persons were enrolled in the controlgroup.The sensitivity and specificity correlations of sB7-H3 in acute pancreatitis with severity degree , as well as with the clinical detection index , were also evaluated. Results The level of plasma sB7-H3 at 24 h in the AP group was significantly higher than that in the healthy control ( HC) group (t = 3.925, P = 0.0002), however, no significant difference was found between the MAP groep and the HC group (P>0.05). The levels of plasma sB7-H3 in the MSAP and the SAP group were significantly higher than that in the HC group (P<0.05和P<0.001)or the MAP group (P<0.05 和P<0.01);The level of plasma sB7-H3 in the SAP group was also markedly higher than that in the MSAP group (P < 0.01). sB7-H3 had a linear positive correlation with LDH、hs-CRP、WBC(P<0.05). ALB had a linear negative correlation with and Ca (S)(P<0.05). By the cutoff of sB7-H3, the sensitivity and specificity to judgethe above moderate pancreatitis were 88.9%and 83.3%,and to judgethe SAP were 96%and 96%. Conclusion sB7-H3 has important clinical value to judge the severity of acute pancreatitis at early time with high sensitivity and specificity , with a linear correlation with the clinical severity index.

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