1.CCN3 inhibits deposition of extracellular matrix of glomerular mesangial cells via promoting the expression of microRNA-29a
Hong LIU ; Haifei LIU ; Long CHEN ; Yi WEN ; Bicheng LIU
Chinese Journal of Nephrology 2017;33(7):531-537
Objective To investigate the effects of nephroblastoma over-expressed protein (CCN3) on the formation of extracellular matrix (ECM) induced by transforming growth factor-β1 (TGF-β1) in human mesangial cells (HMCs) and its underlying signal transduction mechanism related with microRNA-29(miRNA-29).Methods HMCs were pretreated with different doses of exogenous CCN3 (5 μg/L,50 μg/L and 500 μg/L) or transfected with pcDNA3.1(+)-CCN3 before exposed to TGF-β1(2 μg/L),to observe the expression of fibronectin (FN),type I collagen (COL I) and miRNA-29a,b and c.The mimics or inhibitor of the miRNA-29a were transfected into HMCs to analyze whether miRNA-29a affect CCN3.The expressions of FN mRNA,COL I mRNA and miRNA-29 family were detected by real time PCR.The protein expressions of FN and COL I were detected by Western blotting and cell immunofluorescence.Results (1) Compared with the normal control group,the expressions of FN and COL I were up-regulated in TGF-β1 group,while the expressions of miRNA-29a,b,c were down-regulated in TGF-β1 group (all P < 0.05).(2) Compared with the TGF-β1 group,the expressions of FN and COL I were decreased when pretreated with the different doses of exogenous of CCN3 or transfected with pcDNA3.1(+)-CCN3 (all P < 0.05).Meanwhile,the expression of miRNA-29a was significantly increased when pretreated with 50 μg/L and 500 μg/L CCN3 or transfected with pcDNA3.1(+)-CCN3 (all P < 0.05);whereas miRNA-29b and c had no statistical difference (all P > 0.05).(3) Compared with TGF-β1+CCN3 group,the expressions of FN and COL I were decreased in CCN3+TGF-β1+miRNA-29a mimics group (all P < 0.05),whereas the expressions of FN and COL I in CCN3+TGF-β1+miRNA-29a inhibitors group were increased (all P < 0.05).Conclusions CCN3 reduces the TGF-β1-induced production of ECM by the up-regulation of miRNA-29a.
4.Evaluation of Statistical Methods for Articles Published in《Chinese Circulation Journal》
Huanhuan LIU ; Long ZHOU ; Xiaoxiao WEN ; Yaguang PENG ; Liancheng ZHAO
Chinese Circulation Journal 2017;32(3):284-287
Objective: To evaluate the applications and problems of statistical methods for articles published in《Chinese Circulation Journal》. Methods: We reviewed the statistical methods for original articles published in《Chinese Circulation Journal》in 2010, 2011 and 2015; general status with problems was analyzed and the situations were compared among different years. Results: A total of 404 articles were enrolled, statistical hypothesis testing was performed in 366 of them, 32 had descriptive statistics only and 6 articles were meta-analysis. The most frequently used statistical methods were student'st-test (57.4%), followed by chi-squared analysis (56.7%), analysis of variance (35.2%) and regression analysis (23.0%). The main statistical problems in Methods included in described method un-matched to really used method and insufifcient or wrong description. The proportion of insufifcient or wrong description in 2010 and 2011 was 36.6% (71/194), while in 2015 was 26.2% (55/210),P=0.02. The major wrong application of statistical approach was ignoring design with improper method and using unsatisifed precondition for speciifc analysis. The proportion of improper statistical method application in 2010 and 2011 was 19.1% (33/173), in 2015 was 19.7% (38/193),P=0.88. Conclusion: The quality of statistics was improved in 2015 than 2010 and 2011 for articles published in《Chinese Circulation Journal》; while there were still problems at certain degree, enhanced statistical review should be conducted in medical journals.
6.Application of RNA Secondary Structure in Phylogenetic Analysis of Microbiology
Yang LIU ; Xiao-Long CUI ; Wen-Jun LI ; Qian PENG ;
Microbiology 1992;0(02):-
Attention was gradually paid by biologists to the using of RNA secondary structure in the classification of microbiology and phylogenetic relationship analysis in recent years. The development around the research was summarized here briefly. And more emphasis was given to the part introducing the application of RNA secondary structure to the analysis of phylogenetic relationship.
7.Magnetic resonance cholangiopancreatography classification of the bile duct anatomy of liver transplantation living donor: a meta-analysis
Miaomiao LONG ; Lihua LIU ; Tie LIU ; Jianlan YUE ; Wenjun FAN ; Wen SHEN ; Ji QI
Chinese Journal of Organ Transplantation 2012;33(7):407-411
Objective To systematically evaluate the diagnostic value of magnetic resonance cholangiopancreatography (MRCP) in the investigation of bile duct anatomy of liver transplantation living donors.Methods A search in Cochrane library,MEDLINE,EMBASE,CBMdisc (China Biology Medicine disc) was performed to identify relevant English and Chinese-language abstracts,supplemented by Springer,OVID,Sciencedirect full text database,etc.Criteria for inclusion were based on validity criteria for diagnostic research published by the Cochrane collaboration.With Meta analysis package for Stata10.1,heterogeneity of the included articles was tested,which was used to select proper effect model to calculate pooled weighted sensitivity and specificity,positive likelihood ratio,negative likelihood ratio. Summary receiver operating characteristic (SROC) curve was performed and the area under the curve (AUC) was calculated. Finally,sensitivity analysis was performed.Results Seventeen articles with 34 studies were included.Heterogeneity analysis revealed heterogeneity between studies and the source was MRCP imaging methods spotted by meta-regression analysis. Subgroup analysis according to MRCP imaging methods showed homogeneity within subgroups.The pooled sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odd ratio of breath-holding thick slice MRCP,3D MRCP,the combination of the prior two methods,contrast enhance MRCP were 0.89,0.78,4.1,0.14,29; 0.92,0.80,4.5,0.10,45;0.95,0.82,5.2,0.06,85; and 1.00,0.76,4.1,0,1228,respectively with fixed effect model analysis.The area under the SROC curve was 0.83,0.92,0.96 and 0.99 respectively.Conclusion The combination of thick slice and 3D MRCP is a practical and effective method with good sensitivity and specificity to investigate bile duct anatomy of living liver transplantation donors,which fully meets the requirements of the preoperative assessment of bile duct structure.
8.Effect and mechanism of aspirin to inhibit metastasis of colorectal cancer in mice
Wen YUAN ; Qiujuan CHEN ; Xin LIU ; Zihao YANG ; Ping LIU ; Fangyi LONG ; Ting WANG
Journal of Regional Anatomy and Operative Surgery 2017;26(5):319-323
Objective To investigate the inhibitory effect of aspirin on liver metastasis of colon cancer in mice and the possible mechanism.Methods A total of 32 BALB/C mice were injected with CT26 colorectal cancer cells to establish colon cancer liver metastatic model,with 3 mice dead,15 mice in control group and 14 mice in experimental group.The control group was given saline 0.2 mL/d,the experimental group were given aspirin 30 mg/kg.The liver weight and the number of metastatic tumors were calculated after 30 days of intervention.HE and CD31 staining was performed by immunohistochemistry to observe the metastasis and angiogenesis.The protein expression of VEGF and cox-2 were analyzed by Western blot.Results The average liver weight and number of liver metastases nodules in the experimental group were significantly lower than those in the control group(P<0.05).Pathological examination showed that the experimental group of mice the number of liver cells and liver tumor angiogenesis were significantly less than the control group(P<0.05).Western blot showed that the expression of VEGF and cox-2 of CT26 cells were down-regulated after treated with aspirin.Conclusion Aspirin can down regulate the protein expression of VEGF and cox-2 protein to inhibit liver metastasis of colon tumor proliferation and angiogenesis,thereby inhibiting metastasis of colon cancer cells,for therapeutic purposes.
9.Expression of a Novo Human Osteoprotegerin-Mycobacterial Heat Shock 70 Fusion Protein with Baculovirus-Insect Expression System
Shu LIU ; Quan-Geng ZHANG ; Yue ZHANG ; Jing MA ; Zhen-Long LIU ; Wen-Ming ZHAO ;
China Biotechnology 2006;0(05):-
Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P
10.Application of optical coherence tomography in the diagnosis of congenital pseudarthrosis of tibia
Xueqiang NIU ; Yang LIU ; Fuyun LIU ; Weiming HU ; Yuran QU ; Long WEN
Chinese Journal of Applied Clinical Pediatrics 2021;36(7):529-532
Objective:To explore the feasibility of the application of optical coherence tomography (OCT) in the diagnosis of congenital pseudarthrosis of tibia (CPT) in children.Methods:Ten children with neurofibromatosis type Ⅰ (NF1) and CPT were treated in the Third Affiliated Hospital of Zhengzhou University from January 2016 to December 2019, and enrolled as the experimental group.The bone tissue samples were collected and subjected to OCT scanning after intraoperative observation and evaluation, and were contrasted with conventional histological examination.During the same period, the bone tissues of 5 non-NF1 and non-CPT induced-labor fetuses were collected as the control group for the above examination, and the bone tissue examination results of the experimental group and the control group were compared as well.Results:Compared with the bone tissues of the control group, that of the experimental group displayed thickening trabecular bone, part of trabecular bone fusion, disordered arrangement, proliferation and fatification of interstitial fibrous tissue.OCT scan can directly show the nerves and blood vessels in the bone tissue, scattered in adipose tissue.OCT scan showed that the bone tissues of the control group were neatly arranged, with dense and regular shadows.The bone tissues of the experimental group could be observed with strong refraction, loose arrangement, and disordered bone fractures, fibrous ossification, scattered nerves, blood vessels, and increased vacuolar fat cells.Conclusions:OCT can quickly and clearly scan the freshly isolated tissues and directly display the internal structure of the tissues.It is highly compatible with routine pathological examinations and can be an effective supplement to the pathological diagnosis of children with CPT and retain samples for subsequent genetic studies.