1.Stereotaxic intracerebral transplantation of neural stem cells with Nogo-66 receptor gene silencing for treating traumatic brain injury in rats
Dong WANG ; Jianjun ZHANG ; Jingjian MA
Chinese Journal of Tissue Engineering Research 2010;14(14):2539-2544
BACKGROUND:Neural stem cells(NSCs)have the potential of self-proliferation and multiple directional differentiation,and can differentiate into various cells in the neural system under a certain condition.Therefore,NSCs have good prospect in repair of nerve injury.However,RNA interference avoids the abuse of permanent gene silencing,and is hopeful to combine with NSC transplantation for treating craniocerebral injury.OBJECTIVE:To determine whether the Nogo-66 receptor(NgR)gene silencing in NSCs can enhance curative effects of stereotaxic intracerebral transplantation of NSCs on traumatic brain injury(TBI)in rats.METHODS:A total of 60 male Wistar rats following TBI establishment were randomly assigned to 3 groups(n = 20).In the experimental group,NgR gene silencing NSC suspension(6 μL)was injected into rat brain tissue following 24 hours of model induction.In the control group,an equal volume of NSC suspension was infused by the same method.In the blank group,an equal volume of medium without stem cells was infused by the same method.At 24 hours,3 days,1 and 2 weeks following injury,neurological deficits were scored.Two weeks later,animals were sacrificed and subjected to immunohistochemistry and hematoxylin-eosin staining.RESULTS AND CONCLUSION:Following transfection of small interfering RNA,compared with control group,NgR gene protein expression was significantly reduced in the experimental group.At 1 and 2 weeks following transplantation,neurological deficit score was significantly leas in animals undergoing NSC transplantation in the experimental group compared with the control group(P<0.05).Moreover,neuron number in the brain tissue sections of experimental group was significantly more than in the control group(P<0.01).At 2 weeks following injury,hematoxylin-eosin staining showed that brain tissue breakage at damaged site as scar connection,remarkable porosis in the blank group;typical morphological changes as neural cells at the transplanted site in the control group;typical morphological changes as neural cells without cavity in the experimental group.Immunohistochemistry showed(37.92±16.02)BrdU-labeled positive cells/high-power field in the blank group,(89.68±15.34)cells/high-power field in the control group,and(102.67±13.52)cells/high-power field in the experimental group.Significant differences were detected between groups(P<0.01).The above-mentioned results indicated that the NSCs of NgR gene silencing transplanted into the injured cerebral tissues can significantly improve the neurological function in rats with TBI.
2.Control study on MRI of articular cartilage erosion and its histological expression
Baolin ZHU ; Jianhong QI ; Jingjian ZHANG
Orthopedic Journal of China 2006;0(03):-
[Objective]To Study the sensitive sequence of MR for all stages of articular cartilage erosion and to evaluate the effect of MR on the correct diagnosis in the early stage of articular cartilage.[Method]One human fresh amputated knee joint and 4 normal pig knee joints were used to study and select the best sequence by T2WI,PDWI,GE,STIR,3D FS-FSPGR sequences and 6 pig oatcoarthritis(OA)model knee joints were studied by the selected best sequence,MRI was compared with correspont histological evidence to evaluate the effect of MR on the correct diagnosis of OA cartilage.[Result]The best results of tissue resolution and diagnosis rate were seen in 3D FS-FSPGR of all five sequences respectively 67.9% and 93.7%,of proteogly(PG)and collagen fiber(CF)mainly distributed in the deep cantilage.Dyeing of PG in diffent cantilage erosion was decreased while CF increasing.[Conclusion]3D FS-FSPGR is a favorable scan sequence to examine the OA changes of articular cartilage.MRI could show the pathological changes including early stage and have good veracity to the early pathological changes of OA cartilage.
3.Fasudil and bone marrow mesenchymal stem cell transplantation for the treatment of spinal cord injury: Is there a synergistic effect
Jianjun ZHANG ; Dong WANG ; Zepeng LIU ; Jingjian MA
Chinese Journal of Tissue Engineering Research 2009;13(45):8901-8905
BACKGROUND:Research has been proved that Fasudil,a Rho kinase inhibitor,can effectively inhibit the onset of secondary spinal cord injury.OBJECTIVE:To investigate the synergistic effect of bone marrow masenchymal stem cell (BMSC) transplantion and Fasudil on motor functional recovery following spinal cord injury.DESIGN,TIME AND SETTING:A randomized controlled animal experimant was performed at institute of Endocrinology,Tianjin Medical University from November 2008 to March 2009.MATERIALS:A 1-month-old SD rat was obtained to extract BMSCs.Another 30 healthy female SD rats were used to establish spinal cord injury models,and they were then randomly divided into single injury group,cell transplantation group,and cell transplantation + Fasudil group,with 10 rats for each group.Fasudil was provided by Tianjin Hongri Pharmaceutical Co.,Ltd.METHODS:One week after modeling,spinal cord injury was exposed in the cell transplantation group and cell transplantation+Fasudil group,and 10 μL BMSC suspension was inserted into the injured region.Otherwise,6 hours later rats in the cell transplantation +Fasudil group were treated with an intraperitoneal injection of 10 mg/kg Fasudil,twice a day for one successive week.MAIN OUTCOME MEASURES:Hindlimb motor function was detected using inclined plane test.The Phosphorylated-ERM protein expression was detected by hernatoxylin-eosin staining,pathology and horseradish peroxidase (HRP) nerve trace,and Western blot.RESULTS:Eight weeks after modeling,degree of inclined plane was significantly increased in cell transplantation group and cell transplantation + Fasudil group compared with single injury group (P < 0.05,P < 0.01);while the increased value in the cell transplantation group was significantly greater than cell transplantation + Fasudil group (P < 0.05).Broken myeloid tissue and cavitation were observed in the single injury group;a few of neuraxis-like structures were observed in the cell transplantation group and cell transplantation + Fasudil group,but the cavity in the cell transplantation group was larger than cell transplantation + Fasudil group.HRP-pesitive nerve fibers were detected at T_8 segment or even above in the single injury group and increased in cell transplantation group,in particular in cell transplantation + Fasudil group.Phospho-ERM protein expression in the single injury group and cell transplantation group was significantly greater than cell transplantation + Fasudil group (P < 0.05).CONCLUSION:BMSC transplantation can promote hindlimb motor functional recovery following spinal cord injury,while the combined application of cell transplantation and Fasudil may cause a synergistic effect.
4.Investigation and analysis on the quality of teaching bone marrow smear
Rong YU ; Lihua ZHAO ; Fengling QIAO ; Ping LENG ; Chaonan TIE ; Xinyu WU ; Fugui DENG ; Jingjian ZHANG
International Journal of Laboratory Medicine 2017;38(2):172-173,176
Objective To Investigation the quality of marrow smear purchased by CDUTCM.Methods The quality and the typ-icality of marrow smears purchased during 2015 -2016 were collectively examined,and then decided whether these smears fit the blood cell morphology experimental teaching requirement.Results Of all the 960 marrow smears purchased these two years, 49.7% failed in smear made or stained,and 16.0% failed to meet the teaching requirements in the typicality of marrow cells.Con-clusion Teaching marrow smears,being different from clinic ones in their preparation and morphological diagnosis,must be of great quality in sustaining and of better typicality in their cell features.
5. The impact of elastic chest belt on chest wall deformity after costal cartilage harvesting
Jingjian HAN ; Weiwei DONG ; Ying LIU ; Rui GUO ; Ling ZHANG ; Yupeng SONG ; Lin LIN ; Qinghua YANG
Chinese Journal of Plastic Surgery 2018;34(3):210-212
Objective:
To evaluate whether restricted breathing movement by wearing elastic chest belt within half a year postoperatively could decrease the incidence of chest wall deformity after costal cartilage harvesting.
Methods:
The contour of chests of the patients receiving the third-stage operation of auricular reconstruction was assessed 1 year after costal cartilage harvesting with history collection and physical examination. The incidence of costal wall deformity was summarized and compared by wearing elastic chest belt or not within half a year postoperatively. Logistic regression was used to analyze the significance of results.
Results:
Thirty-eight eligible patients were enrolled in the study. The incidence of chest wall deformity in patients wearing chest elastic chest belt as requested for half a year was lower (64.7% vs. 95.2%), and the difference was statistically significant, OR 10.9, 95%CI[1.16, 102.60],
6.Application of pectoralis major flap in complex defects after maxillofacial " frozen neck" surgery
Jingjian WEI ; Hao ZHENG ; Kun HAN ; Xiaodong LI ; Yanhong MA ; Aixia ZHANG ; Jian MENG
Journal of Chinese Physician 2023;25(10):1516-1519
Objective:To summarize the application of pectoralis major flap in complex defects after maxillofacial " frozen neck" surgery.Methods:The clinical data of 7 patients with maxillofacial " frozen neck" admitted to the Department of Stomatology of the Xuzhou Central Hospital from October 2020 to October 2022 were retrospectively analyzed. The pectoralis major flap was used to repair the complex defects after surgery and the treatment effect was observed.Results:All 7 patients had survived flap transplantation with no serious complications. After 3 to 24 months of follow-up, the patients were basically satisfied with the treatment effect.Conclusions:The pectoralis major flap has reliable blood supply, abundant tissue, and can be applied flexibly with a high survival rate and significant repair effect. It is a good choice for repairing complex defects after " frozen neck" surgery, and it has clinical application value.
7.Low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio predicts asymptomatic carotid plaques and their stability in high-risk stroke population
Jianyu ZHANG ; Hui SHI ; Huipin CHEN ; Chuantong ZHANG ; Xingjin DONG ; Linji LIU ; Guangxing WANG ; Jingjian WANG ; Zide GUAN ; Xiaoping TIAN ; Jianming HAN ; Ying SHI ; Yi TANG ; Mingli HE
International Journal of Cerebrovascular Diseases 2019;27(2):104-112
Objective To investigate the relationship between low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio (LHR) and asymptomatic carotid plaques and their stability in high-risk stroke population.Methods Between December 2012 and April 2015,a total of 39 944 permanent resident population ≥40 years were used as subjects of the survey from 11 rural communities in Haitou Town,Banzhuang Town and Tashan Town,Ganyu District,and 9 urban communities in Xinpu District and Haizhou District,Lianyungang City using epidemiological survey method of cluster sampling.Excluding those who took lipid-lowering drugs within 3 months and had a history of stroke or transient ischemic attack,6 592 people at high risk of stroke were finally screened out.Ultrasound was used to detect carotid plaques.The subjects were divided into plaque-free group and plaque group.The latter was further divided into stable plaque group and unstable plaque group.Multivariate logistic regression analysis was used to evaluate the independent risk factor for carotid plaques and their stability.The odds ratio (OR) and 95% confidence interval (CI) were calculated.Receiver Operating Characteristic (ROC) curve was used to evaluate the prediction efficiency of LHR on carotid plaques.Results Multivariate logistic regression analysis showed that low-density lipoprotein cholesterol (LDL-C) was an independent risk factor for carotid plaques,while high-density lipoprotein cholesterol (HDL-C) was an independent protection factor of carotid plaques.Using the lowest quintile (Q1) of LHR as a reference,carotid plaque risk increased significantly with the increasing LHR (Q2:OR 1.448,95% CI 1.082-1.937,P =0.013;Q3:OR 2.414,95% CI 1.754-3.322,P<0.001;Q4:OR 2.939,95% CI 1.945-4.441,P<0.001;Q5:OR 4.884,95% CI 3.143-7.115,P<0.001).ROC curve analysis showed that the area under the curve (AUC) of LHR predicting carotid plaques was 0.795 (95% CI 0.792-0.807;P< 0.001),and the optimal cut-off value was 3.00 (sensitivity 68.37%,specificity 75.65%).LHR ≥3.92 (LHR in the Q4 and Q5 subgroups) was an independent risk factor for unstable carotid plaques (OR 2.915,95% CI 2.104-4.040;P<0.001).The AUC of the LHR predicting unstable carotid plaques was 0.658 (95% CI 0.633-0.684;P<0.001).Conclusions LHR was an independent predictor of carotid plaques in high-risk stroke patients.It had higher predictive value for carotid plaques,and its conversion threshold for promoting plaque formation was 3.00.When LHR was ≥3.92,there was a significant increase in the risk of unstable carotid plaques.
8.Extraction of exosome by gel electrophoresis microfluidic chip and determination of miRNA-21 in exosome of human plasma.
Dan LUO ; Fengying RAN ; Lun WU ; Juan ZHANG ; Fangling REN ; Jingjian LIU ; Binqiang ZHANG ; Qinhua CHEN
Chinese Journal of Biotechnology 2021;37(2):663-672
We developed a high-efficiency microfluidic chip for extracting exosomes from human plasma. We collected peripheral blood from normal human, designed and fabricated a microfluidic chip based on nanoporous membrane and agarose gel electrophoresis to isolate exosomes. The extracted exosomes were characterized by transmission electron microscopy, nanosight and Western blotting, the morphology, concentration and particle size of exosomes were identified and analyzed. Meanwhile, we used ultracentrifugation and microfluidic chip to isolate exosomes separately. The particle size and concentration of the exosomes extracted by two methods were compared and analyzed, and their respective extraction efficiency was discussed. Finally, the expression level of miRNA-21 in exosomes was analyzed by RT-PCR. The microfluidic chip isolated (in 1 hour) high-purity exosomes with size ranging from 30-200 nm directly from human plasma, allowing downstream exosomal miRNA analysis. By comparing with ultracentrifugation, the isolation yield of microfluidic chip was 3.80 times higher than ultracentrifugation when the volume of plasma sample less than 100 μL. The optimized parameters for exosome isolation by gel electrophoresis microfluidic chip were: voltage: 100 V; concentration of agarose gel: 1.0%; flow rate of injection pump: 0.1 mL/h. The gel electrophoresis microfluidic chips could rapidly and efficiently isolate the exosomes, showing great potential in the research of exosomes and cancer biomarkers.
Exosomes
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Humans
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MicroRNAs/genetics*
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Microfluidics
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Plasma
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Ultracentrifugation