1.Decreased c-kit and miR-21 gene expression in rat heart failure
Zhen LIU ; Shuo LI ; Lingling LIU ; Zhikun GUO ; Pengfei WANG
Basic & Clinical Medicine 2017;37(1):76-79
Objective To explore mechanism ( s) is dynamic expression and internal relations of c-kit gene and miR-21 in rat ventricular remodeling .Methods SD rats( n=140) were randomly divided into normal control group ( n=15 ) and heart failure model group ( n=125 ) .Heart failure model:4 mg/kg intraperitoneal injection of adria-mycin.To detect the cardiac function of rats in eight weeks , proving the heart failure .Then harvest the rat heart , frozen section , using immunohistochemistry and immunofluorescence color to detect the expression changes of miR-21 and c-kit in myocardial tissue .Results The emergence of the pathological changes of cardiac muscle cells after myocardial infarction in the heart failure groups and the control group didn't appear the myocardial infarction and heart failure .Immunohistochemistry and immunofluorescence show that miR-21 positive cells in normal and heart failure myocardium mainly express in vascular endothelium ,a few myocardial cells and stem cells , and endocardial expressing quantity fell more than epicardial;c-kit positive cells tend to cluster together , mainly gather in the epi-cardial and its nearby , and the expressing quantity decrease significantly after heart failure .A small number of cells exsit miR-21 and c-kit common expression in both groups ( P<0.05 ) .Conclusions The decreased expression of c-kit and miR-21 is highly related to rats heart failure and left ventricular remodeling .
2.Evidence-based Study on Probiotics for Ulcerative Colitis
Zhikun SHEN ; Zhen WANG ; Jingjing HE ; Miaomiao DONG
China Pharmacy 2001;0(08):-
OBJECTIVE:To evaluate the efficacy of Probiotics in the treatment of ulcerative colitis(UC).METHODS:The recent pertinent literature about the use of Probiotics in the treatment of UC was retrieved from PubMed and Medline and the methodology and literature quality were evaluated in accordance with the evaluation criteria for the quality of literature of evidence-based medicine.RESULTS & CONCLUSION:The majority of the studies showed that Probiotics are of positive value for the remission of the US at acute stage and maintaining of remission induction of drugs in the treatment of UC.Strictly designed and randomized controlled trials(RCTs)remain to be done to tackle the problems encountered by the micro-ecological preparations as a new therapy in the treatment of UC.
3.Differentiation character of adult mesenchymal stem cells and transfection of MSCs with lentiviral vectors.
Xiayi, ZHANG ; Jinsong, LI ; Jun, NIE ; Ke, JIANG ; Zhikun, ZHEN ; Jianjun, WANG ; Lin, SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(6):687-93
This study examined the differentiation character and pluripotency of mesenchymal stem cells (MSCs) under different conditions. Adult MSCs were initially isolated from the bone marrow of rats, cultured in vitro and identified by flow cytometry. After MSCs were transferred to osteogenic and adipogenic medium respectively, the morphological characterization of induced cells was observed. The expression of marker genes was detected by RT-PCR analysis. Then MSCs were transfected with lentiviral vectors pGC-FU-Sox9-EGFP. Enhanced green fluorescence protein (EGFP) expression and transfection efficiency were determined by fluorescence microscopy. The results demonstrated that EGFP caused no effect on the multilineage potential of adult MSCs. Sox9 gene expression of high level was maintained stable in the transfected MSCs and induced MSCs to differentiate into chondrocytes. Aggracan was positive in chondrogenic lineages and the expression of aggracan and type collagen II was significantly increased during MSCs chondrogenic differentiation. It was concluded that Sox9 gene-modified adult MSCs may be promising candidate cells for further studies on tissue engineering. EGFP facilitates the research on MSCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo.
4.Curative effect of intra-tendon sheath hyaluronidase injection guided by ultrasound in treating biceps brachii longitendosynovitis
Zhen HUANG ; Guisheng LIANG ; Zhikun WANG ; Yinguang YANG
Journal of Chinese Physician 2021;23(12):1795-1799
Objective:To observe the effect of hyaluronidase injection into tendon sheath under muscle bone ultrasound guidance in the treatment of tenosynovitis of long head of biceps brachii.Methods:134 patients with tenosynovitis of the long head of biceps brachii treated in SSL Central Hospital of Dongguan from April 2019 to April 2020 were selected as the research objects. All patients were randomly divided into ordinary injection group and ultrasound-assisted injection group, 67 cases in each group. The general injection group was treated with intra-articular injection of hyaluronidase, and the ultrasound-assisted injection group was treated with intra-tendon sheath injection of hyaluronidase under the guidance of muscle and bone ultrasound. Visual Analog Scales (VAS) score, active flexion joint motion (AFROM), shoulder range of motion (ROM), functional score, flexion strength score, nuclear magnetic resonance imaging (MRI) and local tenosynovitis of the long head of biceps brachii (TLHBB)were measured to evaluate the clinical effect and postoperative complications after treatment.Results:There was no significant difference in gender, age, course of disease, periarthritis of shoulder and disuse atrophy of muscles around shoulder between ordinary injection group and ultrasound-assisted injection group ( P>0.05). After treatment, the AFROM, ROM, function score, forward flexion strength score, and middle wedge angle (MWA) of the two groups were significantly higher than those before treatment ( P<0.05), while the VAS score, humeral head diameter (HHD), biceps long head tendon diameter (BTD), and TLBBB were significantly lower than those before treatment ( P<0.05). The AFROM, ROM, function score, and forward flexion strength score, MWA of the ultrasound-assisted injection group were significantly higher than those of the ordinary injection group ( P<0.05), and the VAS score, HHD, BTD, and TLHBB were significantly lower than those of the ordinary injection group ( P<0.05). The total effective rate of the ultrasound-assisted injection group was higher than that of the ordinary injection group (97.01% vs 85.07%, P<0.05). Conclusions:Intra-tendon sheath hyaluronidase injection guided by ultrasound can effectively treat tenosynouitis of the long head of biceps brachii, relieve shoulder pain and improve shoulder motion.
5.Differentiation character of adult mesenchymal stem cells and transfection of MSCs with lentiviral vectors.
Xiayi ZHANG ; Jinsong LI ; Jun NIE ; Ke JIANG ; Zhikun ZHEN ; Jianjun WANG ; Lin SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(6):687-693
This study examined the differentiation character and pluripotency of mesenchymal stem cells (MSCs) under different conditions. Adult MSCs were initially isolated from the bone marrow of rats, cultured in vitro and identified by flow cytometry. After MSCs were transferred to osteogenic and adipogenic medium respectively, the morphological characterization of induced cells was observed. The expression of marker genes was detected by RT-PCR analysis. Then MSCs were transfected with lentiviral vectors pGC-FU-Sox9-EGFP. Enhanced green fluorescence protein (EGFP) expression and transfection efficiency were determined by fluorescence microscopy. The results demonstrated that EGFP caused no effect on the multilineage potential of adult MSCs. Sox9 gene expression of high level was maintained stable in the transfected MSCs and induced MSCs to differentiate into chondrocytes. Aggracan was positive in chondrogenic lineages and the expression of aggracan and type collagen II was significantly increased during MSCs chondrogenic differentiation. It was concluded that Sox9 gene-modified adult MSCs may be promising candidate cells for further studies on tissue engineering. EGFP facilitates the research on MSCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo.
Adult Stem Cells
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cytology
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Aggrecans
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metabolism
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Animals
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Bone Marrow Cells
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cytology
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Cell Differentiation
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Cells, Cultured
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Chondrocytes
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cytology
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Collagen Type II
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metabolism
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Genetic Vectors
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genetics
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Green Fluorescent Proteins
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genetics
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Lentivirus
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genetics
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metabolism
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Mesenchymal Stromal Cells
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cytology
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Rats
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SOX9 Transcription Factor
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genetics
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Tissue Engineering
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Transfection
6.Construction and application value of prognosis associated miRNA prediction model based on bioinforma-tics analysis in pancreatic cancer patients
Jiangning GU ; Haifeng LUO ; Chenqi WANG ; Zhen NING ; Jian DU ; Chi MA ; Yunlong CHEN ; Shimeng CUI ; Zhikun LIN ; Yiping LIU ; Guang TAN
Chinese Journal of Digestive Surgery 2020;19(4):421-430
Objective:To construct a prognosis associated micro RNA(miRNA) prediction model based on bioinformatics analysis and evaluate its application value in pancreatic cancer patients.Methods:The retrospective cohort study was conducted. The clinicopathological data of 171 pancreatic cancer patients from the Cancer Genome Atlas (TCGA) (https: //cancergenome.nih.gov/) between establishment of database and September 2017 were collected. There were 93 males and 78 females, aged from 35 to 88 years, with a median age of 65 years. Of the 171 patients, 64 had complete clinicopathological data. Patients were allocated into training dataset consisting of 123 patients and validation dataset consisting of 48 patients using the random sampling method, with a ratio of 7∶3. The training dataset was used to construct a prediction model, and the validation dataset was used to evaluate performance of the prediction model. Nine pairs of miRNA sequencing data (GSE41372) of pancreatic cancer and adjacent tissues were downloaded from Gene Expression Omnibus database. The candidate miRNAs were selected from differentially expressed miRNAs in pancreatic cancer and adjacent tissues for LASSO-COX regression analysis based on the patients of training dataset. A prognosis associated miRNA prediction model was constructed upon survival associated miRNAs which were selected from candidate differentially expressed miRNAs. The performance of prognosis associated miRNA prediction model was validated in training dataset and validation dataset, the accuracy of model was evaluated using the area under curve (AUC) of the receiver operating characteristic curves and the efficiency was evaluated using the consistency index (C-index). Observation indicarors: (1) survival of patients; (2) screening results of differentially expressed miRNAs; (3) construction of prognosis associated miRNA model; (4) validation of prognosis associated miRNA model; (5) comparison of clinicopathological factors in pancreatic cancer patients; (6) analysis of factors for prognosis of pancreatic cancer patients; (7) comparison of prediction performance between prognosis associated miRNA model and the eighth edition TNM staging. Measurement data with normal distribution were represented as Mean± SD, comparison between groups was analyzed by the student- t test, and comparison between multiple groups was analyzed by the AVONA. Measurement data with skewed data were represented as M (range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Ordinal data were analyzed using the rank sum test. Correlation analysis was conducted based on count data to mine the correlation between prognosis associated miRNA model and clinicopathological factors. COX univariate analysis and multivariate analysis were applied to evaluate correlation with the results described as hazard ratio ( HR) and 95% confidence interval ( CI). HR<1 indicated the factor as a protective factor, HR>1 indicated the factor as a risk factor, and HR equal to 1 indicated no influence on survival. The Kaplan-Meier method was used to draw survival curve and calculate survival rates, and the Log-rank test was used for survival analysis. Results:(1) Survival of patients: 123 patients in the training dataset were followed up for 31-2 141 days, with a median follow-up time of 449 days. The 3- and 5-year survival rates were 16.67% and 8.06%. Forty-eight patients in the validation dataset were followed up for 41-2 182 days, with a median follow-up time of 457 days. The 3- and 5-year survival rates were 15.63% and 9.68%. There was no significant difference in the 3- or 5-year survival rates between the two groups ( χ2=0.017, 0.068, P>0.05). (2) Screening results of differentially expressed miRNAs. Results of bioinformatics analysis showed that 102 candidate differentially expressed miRNAs were selected, of which 63 were up-regulated in tumor tissues while 39 were down-regulated. (3) Construction of prognosis associated miRNA model: of the 102 candidate differentially expressed miRNAs, 5 survival associated miRNAs were selected, including miR-21, miR-125a-5p, miR-744, miR-374b, miR-664. The differential expression patterns of pancreatic cancer to adjacent tissues were up-regulation, up-regulation, down-regulation, up-regulation, and down-regulation, respectively, with the fold change of 4.00, 3.43, 3.85, 2.62, and 2.35. A prognostic expression equation constructed based on 5 survival associated miRNAs = 0.454×miR-21 expression level-0.492×miR-125a-5p expression level-0.49×miR-744 expression level-0.419×miR-374b expression level-0.036×miR-664 expression level. (4) Validation of prognosis associated miRNA model: The C-index of prognosis associated miRNA model was 0.643 and 0.642 for the training dataset and validation dataset, respectively. (5) Comparison of clinicopathological factors in pancreatic cancer patients: results of COX analysis showed that the prognosis associated miRNA model was highly related with pathological T stage and location of pancreatic cancer ( Z=45.481, χ2=10.176, P<0.05). (6) Analysis of factors for prognosis of pancreatic cancer patients: results of univariate analysis showed that pathological N stage, radiotherapy, molecular targeted therapy, score of prognosis associated miRNA model were related factors for prognosis pf pancreatic cancer patients ( HR=2.471, 0.290, 0.172, 2.001, 95% CI: 1.012-6.032, 0.101-0.833, 0.082-0.364, 1.371-2.922, P<0.05). Results of multivariate analysis showed that molecular targeted therapy was an independent protective factor for prognosis of pancreatic cancer patients ( HR=0.261, 95% CI: 0.116-0.588, P<0.05) and score of prognosis associated miRNA model≥1.16 was an independent risk factor for prognosis of pancreatic cancer patients ( HR=1.608, 95% CI: 1.091-2.369, P<0.05). (7) Comparison of prediction performance between prognosis associated miRNA model and the eighth edition TNM staging: in the training dataset, there was a significant difference in the prediction probability for 3- and 5-year survival of pancreatic cancer patients between prognosis associated miRNA model and the eighth edition TNM staging ( Z=-1.671, -1.867, P<0.05). The AUC of the prognosis associated miRNA model and the eight edition TNM staging for 3- and 5-year survival prediction was 0.797, 0.935 and 0.737 , 0.703, with the 95% CI of 0.622-0.972, 0.828-1.042 and 0.571-0.904 , 0.456-0.951. The C-index was 0.643 and 0.534. In the validation dataset, there was a significant difference in the prediction probability for 3- and 5-year survival of pancreatic cancer patients between prognosis associated miRNA model and the eighth edition TNM staging ( Z=-1.729, -1.923, P<0.05). The AUC of the prognosis associated miRNA model and the eight edition TNM staging was 0.750, 0.873 and 0.721 , 0.703, with the 95% CI of 0.553-0.948, 0.720-1.025 and 0.553-0.889, 0.456-0.950, respectively. The C-index was 0.642 and 0.544. Conclusions:A prognosis associated miRNA prediction model can be constructed based on 5 survival associated miRNAs in pancreatic cancer patients, as a complementation to current TNM staging and other clinicopathological parameters, which provides individual and accurate prediction of survival for reference in the clinical treatment.