1.Human Umbilical Cord Mesenchymal Stem Cells Improve the Necrosis and Osteocyte Apoptosis in Glucocorticoid-Induced Osteonecrosis of the Femoral Head Model through Reducing the Macrophage Polarization
Gang TIAN ; Chuanjie LIU ; Qi GONG ; Zhiping YU ; Haitao WANG ; Daoqiang ZHANG ; Haibo CONG
International Journal of Stem Cells 2022;15(2):195-202
Background and Objectives:
Apoptosis is an outstanding determinant of glucocorticoid (GC)-induced osteonecrosis of the femoral head (ONFH). Human umbilical cord mesenchymal stem cells (hUC-MSCs) have been demonstrated to be associated with apoptosis in diseases models. However, the role of hUC-MSCs in GC-induced ONFH via regulating apoptosis still needs further study.
Methods:
and Results: In the present study, a GC-induced ONFH model was built in vivo through a consecutive injection with lipopolysaccharide (LPS) and methylprednisolone. The necrosis and apoptosis of the femoral head was evaluated by histological and Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling (TUNEL) assay. The level of collagen and TRAP positive cells were determined by Masson and TRAP staining, respectively. M1 macrophage polarization was assessed using immunofluorescence assay. The level of proinflammatory cytokines including tumor necrosis factor (TNF)‐α, Interleukin (IL)‐1β and IL-6 of femoral head was determined by enzyme-linked immunosorbent assay (ELISA) kits. The protein expression of AKT, mTOR, p-AKT and p-mTOR was detected using western blot assay. The results showed that hUC-MSCs treatment prominently promoted the GC-induced the decrease of the collagen level and the increase of TRAP positive cells. Besides, hUC-MSCs treatment decreased necrosis and apoptosis, macrophage polarization, the level of TNF‐α, IL‐1β and IL-6, the protein expression of p-AKT and p-mTOR, and the radio of p-AKT to AKT and p-mTOR to mTOR of femoral head in vivo.
Conclusions
Therefore, the present study revealed that hUC-MSCs improved the necrosis and osteocyte apoptosis in GC-induced ONFH model through reducing the macrophage polarization, which was associated with the inhibition of AKT/mTOR signaling pathway.
2.Circular RNA HECTD1 participates in oxygen-glucose deprivation-induced neuronal cell damage by regulating miR-98-5p/ephrin A4 expressions
Yifeng LIU ; Changming WEN ; Chuanjie WU ; Jun GAO ; Jun SUN ; Meng SUN ; Shiqian GUO
Chinese Journal of Neuromedicine 2022;21(6):541-552
Objective:To explore whether circular RNA HECTD1 (circ-HECTD1) is involved in oxygen-glucose deprivation (OGD)-induced neuronal cell damage by regulating the expressions of miR-98-5p/ephrin A4 (EPHA4).Methods:Mouse primary cortical neuronal cells were isolated and cultured in vitro. The targeting relations of circ-HECTD1 and miR-98-5p with EPHA4 were detected by dual luciferase reporter assay and RNA binding protein immunoprecipitation assay. These neurons were randomly divided into control group (cultured for 24 h under normal condition) and 6, 12 and 24 h OGD treatment groups (treated with OGD for 6, 12 and 24 h, respectively), OGD+Vector group and OGD+circ-HECTD1 group, OGD+small interfering RNA (siRNA) negative control (si-NC) group and OGD+siRNA circ-HECTD1 (si-circ-HECTD1) group, OGD+micro RNA (miR) negative control (miRNC) group and OGD+miR-98-5p mimic group, OGD+miRNA inhibitor negative control (anti-miRNC) group and OGD+miR-98-5p inhibitor (anti-miR-98-5p) group, OGD+miR-98-5p mimic+pcDNA group and OGD+miR-98-5p mimic+EPHA4 group, OGD+si-circ-HECTD1+anti-miR-NC group and OGD+si-circ-HECTD1+miR-98-5p inhibitor group; pCD5-ciR empty vector, pCD5-ciR-circ-HECTD1, si-NC, si-circ-HECTD1, miR-NC, miR-98-5p mimic, anti-miR-NC or anti-miR-98-5p were transfected into the neurons, and miR-98-5p mimi and pcDNA3.1 empty vector, miR-98-5p mimic and pcDNA3.1-EPHA4 overexpression vector, si-circ-HECTD1 and anti-miR-NC, or si-circ-HECTD1 and anti-miR-98-5p were co-transfected into the neurons. After 24 h of OGD treatment, the circ-HECTD1, miR-98-5p and EPHA4 mRNA expressions were detected by real-time fluorescent quantitative PCR (qRT-PCR), the EPHA4 protein expression was detected by Western blotting, the proliferation activity was detected by MTT assay, the apoptosis rate was detected by flow cytometry, the levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in cell culture medium were detected by ELISA, and the activities of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by kit assay. Results:(1) Targeting relations between circ-HECTD1 and miR-98-5p, and EPHA4 and miR-98-5p were verified. (2) As compared with the control group, the neurons in 6, 12 and 24 h OGD treatment groups had significantly increased circ-HECTD1 and EPHA4 protein expressions and significantly decreased miR-98-5p expression ( P<0.05). (3) As compared with OGD+Vector group, OGD+circ-HECTD1 group had significantly increased circ-HECTD1 expression, and significantly decreased miR-98-5p expression ( P<0.05); as compared with OGD+si-NC group, OGD+si-circ-HECTD1 group had significantly increased miR-98-5p expression, and significantly decreased EPHA4 mRNA and protein expressions ( P<0.05); as compared with OGD+miR-NC group, OGD+miR-98-5p mimic group had significantly increased miR-98-5p expression, and significantly decreased EPHA4 protein expression ( P<0.05); as compared with OGD+anti-miR-NC group, OGD+anti-miR-98-5p group had significantly decreased miR-98-5p expression, and significantly increased EPHA4 protein expression ( P<0.05); as compared with the OGD+si-circ-HECTD1+anti-miR-NC group, OGD+si-circ-HECTD1+anti-miR-98-5p group had significantly increased EPHA4 mRNA and protein expressions ( P<0.05). (4) As compared with the control group, the OGD groups had significantly decreased cell viability and SOD activity, and significantly increased IL-1β and TNF-α levels, apoptosis rate and MDA activity ( P<0.05); as compared with the OGD+si-NC group, the OGD+si-circ-HECTD1 group had significantly decreased cell apoptosis rate, IL-1β and TNF-α levels, and MDA activity, and significantly increased cell viability and SOD activity ( P<0.05); as compared with the OGD+si-circ-HECTD1+anti-miR-NC group, the OGD+si-circ-HECTD1+anti-miR-98-5p group had significantly decreased cell viability and SOD activity, and significantly increased IL-1β and TNF-α levels, apoptosis rate and MDA activity ( P<0.05); as compared with the OGD+miR-NC group, OGD+miR-98-5p mimic group had significantly decreased cell apoptosis rate, IL-1β and TNF-α levels, and MDA activity, and significantly increased cell viability and SOD activity ( P<0.05); as compared with OGD+miR-98-5p mimic+pcDNA group, OGD+miR-98-5p mimic+EPHA4 group has significantly increased cell apoptosis rate, IL-1β and TNF-α levels, and MDA activity, and significantly increased cell viability and SOD activity ( P<0.05). Conclusion:Knockdown of circ-HECTD1 could ameliorate the OGD-induced neuronal cell damage in mice by targeting the expressions of miR-98-5p/EPHA4.
3.Nomogram for predicting Gleason grouping upgrading(GGU)in a cohort receiving radical prostatectomy based on 2014 ISUP grouping system: development and internal validation
Ao LIU ; Hai HUANG ; Chuanjie ZHANG ; Jingyi HUANG ; Yang XU ; Da HUANG ; Rong NA ; Lu CHEN ; Yi GAO ; Danfeng XU
Chinese Journal of Urology 2020;41(4):297-302
Objective:To analyze the predictive factors of GGU between biopsy and radical prostatectomy pathology based on 2014 ISUP grouping system, then establish and evaluate nomogram.Methods:Patients undergoing radical prostatectomy in Shanghai Ruijin Hospital from March 2012 to March 2019 were reviewed, and the clinical and pathological information were collected. Age(68.1±7.2), body mass indes(BMI) (24.2±3.2)kg/m 2, prostate specific antigen(PSA) 11.5(6.7-20.4)ng/ml, prostate specific antigen destiny(PSAD) 0.35(0.20-0.66). Before March 2017, the number of biopsy cores were 6 to 8; After then, all patients toke 12 cores systemic biopsy. Based on 2014 ISUP grouping system, the differences between biopsy and radical prostatectomy grades were counted. The independent predictors of GGU were analyzed by univariate and multivariate logistic regression analysis, then the nomogram for predicting GGU were established and evaluated. Results:429 patients were enrolled. There were 161 (37.5%) patients in GGU group and 268 (62.5%) patients in non-GGU group. After multivariate logistic regression analysis, body mass index (BMI)>28 kg/m 2( OR=2.54, P=0.021), prostate specific antigen density (PSAD)( OR=1.65, P=0.018)and 2014 ISUP grouping sysyem ( OR=0.53, P<0.001) of biopsy specimen were independent impact factors of GGU. The predicting model was established according to BMI, PSAD and 2014 ISUP grouping system. The area under the ROC cure of the model was 0.735 (95% CI 0.681-0.789). The nomogram model was well calibrated, with the mean absolute error of 6.7%, which means the prediction of GGU is fairly consistent with the actual situation. Conclusions:Based on the 2014 ISUP grouping system, BMI>28 kg/m 2, PSAD and 2014 ISUP grouping of biopsy specimen were independent predictors of GGU. The nomogram model for predicting GGU has a good statistical significance.
4.Study on Antibacterial Activities of Meropenem Combined with Cefoperazone Sulbactam to 3 Kinds of Multidrug Resistant Gram-negative Bacteria
Yanlin LIU ; Chuanjie SONG ; Ling YIN ; Xiaoyan LIANG ; Han ZHANG ; Changzheng LI
China Pharmacy 2017;28(21):2912-2915
OBJECTIVE:To study antibacterial activities of meropenem(MPN)combined with cefoperazone sulbactam(SCF) to 3 kinds of multidrug resistance (MDR) Gram-negative bacteria. METHODS:Each 50 strains of MDR-Escherichia coli (MDR-EC),MDR-Klebsiella pneumoniae (MDR-KPN) and MDR-Acinetobacter baumannii (MDR-AB) were isolated from spu-tum,blood,urine,ascites or drainage specimens of patients during Jan. to Dec. in 2016 from the affilidated hospital of Taishan medical university. The agar dilution method and board method were used to determine MIC50,MIC90 and MICG of MPN,SCF, MPN+SCF to MDR-EC,MDR-KPN,MDR-AB and calculate fractional inhibitory concentration (FIC). Drug sensitivity test was conducted by K-B disk method. RESULTS:In terms of the MICG to MDR-EC,MDR-KPN,MDR-AB,MPN alone were respec-tively 36.82,82.45,34.32 μg/mL;SCF alone were respectively 42.14,112.67,24.11 μg/mL;MPN combined with SCF were re-spectively 25.97,56.64,11.36 μg/mL. In terms of MICG to MDR-EC,MDR-KPN,MICG showed that MPN+SCF
5.Efficacy analysis of endovascular treatment of vertebral basilar artery dissecting aneurysms
Lisong BIAN ; Jingwei LI ; Hongqi ZHANG ; Guilin LI ; Chuan HE ; Chuanjie LI ; Lishuang YE ; Jiang LIU ; Zhichao WANG
Chinese Journal of Cerebrovascular Diseases 2016;13(5):257-261
Objective To investigate the effect of endovascular treatment of vertebral basilar artery dissecting aneurysms. Methods The clinical data of 40 patients with vertebral basilar artery dissecting aneurysm admitted to Beijing Xuanwu Hospital and Haidian hospital,Capital Medical University from August 2013 to September 2014 were analyzed retrospectively. Their clinical symptoms and imaging were followed up. According to the treatment methods,they were divided into either a stent-assisted coil emboliza-tion group (group A;n = 34)or a parent artery occlusion (group B;n = 6),and according to the clinical symptoms and imaging followed-up,the efficacy was assessed at 1 year after procedure. Results The patients were followed up for 1 year after procedure,29 patients (85. 3%)were improved in group A, 1 (2.9%)was stable,and 4 (11. 8%)deteriorated. All the 4 deteriorated patients died of cerebral infarction complications (at 1 week to 6 months after procedure). The 6 patients in group B were improved compared with before procedure. No intracranial hemorrhage and ischemic events occurred. Conclusion Using the stent-assisted coil embolization technique and the parent artery occlusion technique for the treatment of the vertebral basilar artery dissecting aneurysms are relatively safe and effective.
6.Analysis of diagnosis and treatment of dural arteriovenous fistula of super petrosal venous drainage
Yongjie MA ; Chuanjie LI ; Lisong BIAN ; Jiang LIU ; Zhichao WANG ; Guilin LI ; Hongqi ZHANG
Chinese Journal of Cerebrovascular Diseases 2016;(2):89-94
Objective To investigate the clinical manifestations and imaging features of dural arterio-venous fistula of super petrosal venous drainage and treatment. Methods From May 2013 to September 2014,9 patients with petrosal vein drained dural arteriovenous fistula at the Department of Neurosurgery, Xuanwu Hospital,Capital Medical University and the Department of Neurosurgery,Beijing Haidian Hospital were enrolled retrospectively. The patients were treated with endovascular embolization or microsurgery,and the MRI and DSA examinations were improved,and the scores of the modified Aminoff&Logue scale (ALS) were performed before and after treatment. Results In the 9 patients,there were 3 females and 6 males. They all had different degrees of limb sensory and motor abnormalities,7 of them also had urination and/or bowel disorders,4 had cranial nerve dysfunction,including hoarseness,bucking,hiccup,and paralysis. Six patients received embolization treatment,3 received microsurgery,and they all achieved anatomic cure. The preoperative ALS score was 6. 0 ± 2. 7,and the score at 3 months after procedure was 2. 8 ± 1. 7. There was significant difference between before and after treatment (t=4. 816,P<0. 05). Conclusions The petrosal vein drained dural arteriovenous fistula is a kind of rare cerebrovascular malformation. The lesion involves a wide range. The clinical manifestations are severe. Both endovascular embolization and microsurgery can achieve a more ideal therapeutic effect. If the vascular condition is permitted,the interventional embolization treatment should be preferred.
7.Influencing factors and the drug purchase behaviors of outpatients in Beijing, Zhengzhou and Guangzhou
Wei LIU ; Feng WU ; Guga SURI ; Chuanjie DUAN ; Xiaoxue ZHANG
Chinese Journal of Health Policy 2015;(10):46-50
Objective:To understand the drug purchase behaviors and prescription flows of outpatients and their influencing factors. Methods: After selecting and investigating tertiary hospitals, secondary hospitals, community health service centers ( one per each level) in Beijing, Zhengzhou, Guangzhou, this paper uses structured question-naire method to analyze 3,155 valid questionnaires. A logistic regression analysis is performed to understand the fac-tors influencing the drug purchase behaviors. Results:The factors influencing the drug purchase behaviors of outpa-tients mainly include the region where the patient is, level of the hospital, type of health insurance that the patient has, frequency of drug purchase, etc. Conclusions: This paper suggests hospital administrators to take measures to improve prescription flows, such as reforming the health care insurance to make payment system more suitable for pharmacies standardizing supply channels in order to make varieties consistent between hospitals and pharmacies, strengthening education and publicity to raise the outpatients’ awareness for drug purchase outside of hospitals, etc.
8.Study on the binding sites of radiosensitivity associated transcription factor in the promoter region of Ier5 gene
Wei CUI ; Lingling YIN ; Lingyue DONG ; Li LI ; Zhicheng LIU ; Yanling LI ; Wei AN ; Xiaodan LIU ; Chuanjie YANG ; Pingkun ZHOU ; Kuke DING
Chinese Journal of Radiological Medicine and Protection 2012;32(1):15-19
Objective To clarify the mechanism of immediate early response gene 5 (ler5)transcription induced by radiation. Methods Deletant construction, site-specific mutagenesis,electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were used to forecast the promoter region,binding sites and transcription factors of Ier5 gene in HeLa cells.Results The promoter region of Ier5 gene might be in the region of Ier5 -8 deletant ( -408 - -238 bp).The Ier5 gene had two transcription factors of GCF and NFI,and GCF had two binding sites located in the region of - 388 - - 382 bp and - 274 - - 270 bp of Ier5 promoter.The binding site of NFI was located in - 362 --357 bp of Ier5 promoter. GCF could inhibit the expression of Ier5 gene and this inhibition was diminished when the radiation dose increased. In contrast, NFI increased the expression of Ier5.Conclusions The most possible region of Ier5 promoter is from -408 to - 238 bp which has two binding sites for the radiosensitivity transcription factors of GCF and NFI that could negatively and positively regulate the expression of Ier5 respectively.
9.Application of anterograde tubular ileal fistula in Ⅰ stage anastomosis of colon cancer with acute obstruction
Shufan JIA ; Chuanjie ZHAO ; Hengling GAO ; Maorong LIU ; Shuliang LI
Chinese Journal of Postgraduates of Medicine 2011;34(20):25-26
Objective To investigate the feasibility of application of anterograde tubular ileal fistula in Ⅰ stage anastomosis of colon cancer with acute obstruction. Method Eighty patients of colon cancer with acute obstruction who treated with anterograde tubular ileal fistula in Ⅰ stage anastomosis were analyzed restropectively. Result Clinical observation showed that 80 patients were cured and discharged,no one did occur anastomotic leakage, abdominal abscess,and other serious complications. Conclusion If correctly graspe the timing of operation for colon cancer with acute obstruction,irrigation methods,good perioperative management,select the anterograde tubular ileal fistula, Ⅰ stage resection and anastomosis is safe and feasible.
10.Construction and characterization of cDNA library for IRM-2 mice
Qin WANG ; Jin LI ; Li SONG ; Qiang LIU ; Jingyin YUE ; Chuanjie MU ; Weisheng TANG ; Feiyue FAN
Chinese Journal of Radiological Medicine and Protection 2010;30(3):274-278
Objective To screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.

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