1.The protective role of peroxisome proliferator activated receptors-α and pathomechanism in D-galactosamine/lipopolysaccharide-induced acute liver failure in rats
Mingjing JIAO ; Feng REN ; Li ZHOU ; Zhongping DUAN ; Caiyan ZHAO
Chinese Journal of Internal Medicine 2014;53(9):730-734
Objective To determine the role and mechanism of peroxisome proliferator activated receptors (PPAR) α in a mouse model of D-galactosamine/lipopolysaccharide (D-GalN/LPS)-induced acute liver failure(ALF).Methods Firstly,C57BL/6 mice were randomly divided into control group(n =8),ALF 2h group(n =8),ALF 4h group (n =8),ALF 6h group (n =8).Secondly C57BL/6 mice were randomly divided into control group(n =8),ALF group(n =8),WY14643 group(n =8).To induce ALF,the mice were injected intraperitoneally with D-GalN (700 mg/kg) and LPS (10 μg/kg).WY14643 (6 mg/kg),the selective agonist of PPAR α,was administered via tail vein two hours prior to D-GalN/LPS exposure.Two,four,and six hours after D-GalN/LPS treatment in the first study,mice were anesthetized and blood was collected,6h after D-GalN/LPS treatment in the second study,blood was collected.The liver tissue was harvested for histology and mRNA extraction.Serum levels of ALT and AST were measured to evaluate the hepatic damage.Inflammatory cytokines (TNFα,IL-1β,IL-6) and chemokines (CXCL-1,CXCL-10) were detected by real-time quantitative PCR.Differential protein expression of p-NF-κBp65,p-JNK,p-ERK,p-p38 in inflammatory pathways was detected by Western blotting.Significance of inter-group differences was assessed by one-way ANOVA,and pairwise comparison was performed by the least significant difference test.Results The gene and protein expression of PPAR α were gradually reduced during the development of ALF.Compared with the model group,the liver architecture was better preserved almost with normal morphology in WY14643-treated mice.Serum ALT and AST levels in WY14643-treated group were significantly lower [ALT:(555 ±62)U/L vs (2 898 ±822) U/L,P <0.05; AST:(791 ±58) U/L vs (3 013 ±997)U/L,P < 0.05].The expression of proinflammatory cytokines and chemokines was significantly suppressed during the activation of PPAR α.In the second study,the levels of gene expression of proinflammatory cytokines and chemokines were detected in control group,ALF group and WY14643 group respectively as followings:TNFα (0.161 ± 0.085,7.996 ± 1.068,3.346 ± 0.94,P < 0.05),IL-1β(0.041 ±0.002,3.657 ±0.904,0.176±0.089,P<0.01),IL-6 (0.018 ±0.008,1.762 ±0.589,0.163±0.0487,P <0.05),CXCL-1 (0.063 ±0.008,7.881 ±0.966,2.737 ±0.864,P <0.01),CXCL-10 (0.054 ±0.005,5.671 ±0.948,2.578 ±0.804,P <0.05).Conclusion Our findings first demonstrate that PPARα protects liver from injury in an ALF mouse model by suppressing inflammatory response,indicating PPARα as a potential new therapeutic target for ALF.
2.The detection and statistical analysis of blood lymphocyte immunophenotype
Wei CUI ; Lingqun ZHU ; Mingjing ZHAO ; Al ET
Chinese Journal of Immunology 2001;0(10):-
Objective:To establish the reference values on blood lymphocyte immunophenotype by means of standardized quality control and proper statistical analysis.Methods:The whole bloods of 93 healthy Chinese adults were stained by fluorescent conjugated monoclonal antibodies of Simultest TM IMK lymphocyte Kit and then were analysed by FACS Calibur flow cytometer.The data were acquired and were analysed by SimulSET.Statistical analysis would be conducted after conforming the date of standardized quality control.Results:CD + 3,CD + 3CD + 4,CD + 3CD + 8 cells showed normal distribution but CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 showed skewness distribution .the 95% reference of confidence interval of CD + 3?CD + 3CD + 4?CD + 3CD + 8 cells were established by the method of normal distribution and they were respectively 49 28%~80 52%?25 29%~48 11%?11 93~44 77%;the 95% reference of confidence interval of CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 were established by the method of percentiles and they were respectively 4 74%~19 84%?10 63%~42 86%?0 65~3 05.Conclusion:To establish the reference values,it is considered that the methods of normal distribution and percentiles should be use comprehensively.It is also suggested that the reference values should be confirmed by means of standardized quality control and proper statistical analysis,so the reference values obtained from various laboratories of the same race but in different regions with the same reagents and instruments performed compatibility.
3.Circulating microRNA-21 in hepatocellular carcinoma patients
Lan GAO ; Huifang LIU ; Mingzhi LING ; Wenqian TIAN ; Mingjing ZHAO ; Wenxin ZHAO ; Poshi XU
Chinese Journal of Laboratory Medicine 2015;(7):484-486
Objective To evaluate the plasma level of miR-21 in hepatocellular carcinoma (HCC) patients and its clinical diagnostic significance .Methods Case-control study was used .The plasma level of miR-21 was measured by real-time quantitative PCR.The relative expressions of miR-21 were calculated. This study included 60 cases of patients with HCC , 71 patients with liver cirrhosis ( LC ) , 52 healthy volunteers ( HV) from January to June in 2014 in Henan Province People′s Hospital.The receiver operating characteristic ( ROC) curves were analyzed to determine the sensitivity and specificity of miR-21 expression levels in HCC diagnosis. Differences between groups were assessed by the t-test.Results Plasma microRNA-21 level in the 60 patients with hepatocellular carcinoma ( 2.6 ±1.1 ) was significantly higher than in patients with chronic hepatitis (1.6 ±0.9) and healthy volunteers (1.0 ±0.6) (t=5.322,P=0.004;t =8.349, P =0.000 3, respectively ) .Plasma microRNA-21 level in the HCC patients were positively correlated with tumor size and differentiation (tdif=3.366,P=0.019;tsize =3.490,P=0.012). ROC analysis of plasma microRNA-21 yielded an AUC of 0.796 with 70.0% sensitivity and 65.3%specificity when differentiating hepatocellular carcinoma from chronic hepatitis , and an AUC of 0.934 with 89.5% sensitivity and 81.8% specificity when differentiating hepatocellular carcinoma from healthy volunteers.Conclusion The plasma level of miR-21 in HCC patients has high specificity , and maybe help to diagnose of HCC.
4.Community-acquired infections and risk factors in patients with diabetes:A Meta-analysis
Guangdan ZHAO ; Mingjing ZHAO ; Fangzhi LI ; Lingling WANG ; Jiaying SUN ; Donghong CHEN ; Xiaoge WANG
Chinese Journal of Infection Control 2014;(10):577-583
Objective To analyze the types and risk factors of community-acquired infections (CAI)in diabetic patients by system analysis method of evidence-based medicine.Methods China National Knowledge Infrastructure (CNKI),Wanfang database,VIP database were searched by computer,domestic published researches on CAI and related risk factors in dia-betic patients were aggregated,Meta-analysis was conducted by stata 1 1 .0 software.Results A total of 1 2 literatures were included in the study .The average rate of CAI in diabetic patients was 39.55% (22.12%-55.86%).The major infec-tions were respiratory system infection(40.74%),urinary tract infection(27.35%),tuberculosis(10.80%),skin and soft tissue infection(9.19%),and hepatobiliary system infection (5.57%).Stratified analysis on risk factors revealed that OR and OR95%CI of chronic complication,age,disease course,glycemic control,gender,type of diabetes,subtype of ketoac-idosis was 1.63(1.45,1.82),1.30(1.19,1.42),1.47(1.35,1.61),0.68(0.61,0.76),0.69(0.64,0.75),1.37 (1.13,1.66 )and 0.87(0.62,1.23),respectively.There was no publication bias and combined results were stable. Conclusion The main CAI in diabetic patients are respiratory system infection,urinary tract infection,tuberculo-sis,skin and soft tissue infection,and so on ;several factors,such as female,older age,long-term disease course, poor glycemic control,and complication,can contribute to the increase of CAI in diabetic patients.
5.The integrin linked kinase promotes migration and invasion of lung cancer cell by upregulating matrix metalloproteinase-9 through nuclear factor-κB pathway
Mingjing ZHAO ; Shuo LIU ; Lingling WANG ; Bing HAN ; Xiaoge WANG ; Xuhua ZHANG
Clinical Medicine of China 2014;30(3):243-248
Objective To investigate the role of integrin-linked kinase (ILK) on migration and invasion of lung cancer cell by upregulating matrix metalloproteinase-9 through nuclear factor-κB (NF-κB) pathway.Methods A549 cell line were overexpressed ILK and matrix metalloproteinase-9 (MMP-9) confirmed by cell transfection,siRNA interference,cell scratch test,real-time quantitative PCR and Western Blot.Results Over-expression of ILK stimulated MMP-9 expression in lung cancer cells(P < 0.01).The addition of MMP-9 inhibitor doxycycline and anti-MMP-9 neutralizing antibody significantly impaired the wound healing capacity of ILK-transfected cells(P < 0.01),as well as by in vitro matrigel invasion assay (P < 0.01).In addition,overexpression ILK induced phosphorylation and nuclear translocation of nuclear factor-κB subunit p65.Upregulation of MMP-9 was severely abolished by either BAY 11-7028,a specific NF-κB inhibitor,or siRNA targeted to NF-κB p65 in ILK over-expression cells.Conclusion The finding indicate that over-expression of ILK can promote the migration and invasion of lung cancer cell,and upregulate MMP-9 through the NF-κB pathway.
6.Survey on smoking status and risk factors in residents
Xiaoyu HE ; Jiaying SUN ; Shuo LIU ; Lingling WANG ; Mingjing ZHAO ; Xiaoge WANG
Chinese Journal of General Practitioners 2012;11(3):213-214
A survey on smoking status and risk factors was conducted among 3373 residents in two districts of Shen yang city during November 2008 and September 2009 ; the pulmonary function tests were also performed for all subjects.A logistical regression model was used to evaluate the risk factors of smoking,and the knowledge about diseases caused by smoking was evaluated by chi-square test.The overall smoking rate was 50.7% ( 1710/3373 ) ; 86.8% (1458/1680) for males a(un)14.9% (252/1693)for females.The risk factors of smoking ( P < 0.05 ) in order of OR value were as follows:drinking,work environment and underlying diseases ; the protective factors were:female,educational level,overweight ( BMI ≥ 24) and old age.The survey on knowledge of smoking-related disease was conducted in 2478 residents including 815 smokers and 1663 non-smokers. The awareness levels about whether smoking can cause COPD,asthma,birth defects,malignant tumor,abortion,growth retardation,fetal death and myocardial infarction were significantly different between smokers and non-smokers.
7.Research of Fusion Expression of Human β-Defensin-3 in Escherichia coli
Yahua ZHAO ; Laixiang XU ; Jianjun ZHANG ; Pengliang HUANG ; Shaopeng XU ; Mingjing SONG
Chinese Journal of Comparative Medicine 2007;17(11):670-675
Objective To achieve the fusion expression of the entire human beta-defensin-3(hBD-3) gene. Method We synthesized two oligonucleotide primers accor ding to the codon preference of Escherichia coli. The gene was cloned into p GEX -4T-2 to establish the pGEX-4T-2-hBD-3 as the fusion expression vector by PCR. Transformed into E.coli strain DH5α, the express vector was induced an d ex pressed by IPTG. The fusion protein GST-hBD-3 was obtained by repeated cycles of freezing and thawing, cut by thrombin to attain the recombinant hBD-3 protei n. Result The result of the antibacterial peptide agarose diffu sion assay shows the antibacterial activity of the rhBD-3 against the S.aureu s exists, and it reaches 0.843U. Conclusion The fusion expr ession of the hBD-3 gene is successful.
8.Effects of Banxia Xiexin Decoction and Its Components on Protein Expression of Cx43 in Gap Junction in Gastric Tissue of Electrogastric Dysrhythmias Rats
Cui LI ; Dongmei ZHANG ; Lixia LOU ; Mingjing ZHAO ; Lina LI ; Meng CHEN
Chinese Journal of Information on Traditional Chinese Medicine 2015;(6):75-79
Objective To discuss compatibility regularity of Banxia Xiexin Decoction adjusting gastric mobility. Methods Ninty healthy SD male rats were divided into normal group (10 rats) and model group (80 rats). Rat models of electrogastric dysrhythmias were established. After the evaluation of gastric electrophysiology, Banxia Xiexin Decoction were divided into Xinkai, Kujiang, Ganbu, Xinkaikujiang, Xinkaiganbu, and Kujiangganbu groups according to the compatibility regularity of Banxia Xiexin Decoction, 10 rats for each group. Rats in each medication administration group received gavage with same concentration but different volume for 4 weeks. Each group was analyzed for gastric electrical parameters after administration. The expressions of Cx43 and Cx43 mRNA in the gastric tissue of rats in each group were analyzed by immunohistochemistry and real-time fluorescent quantitative PCR. Results Compare with the normal group, the expressions of Cx43 and Cx43 mRNA in the gastric tissue of rats increased significantly in the model group. Compared with the model group, variable coefficient of electrogastric slow wave frequency in all medication administration groups decreased obviously. Banxia Xiexin Decoction and its components had decreased the expression of Cx43 and Cx43 mRNA to some extent, among which Xinkaiganbu group had best effects. Conclusion The mechanism of Banxia Xiexin Decoction adjusting gastric motility maybe related to its function of decreasing the expressions of Cx43 and Cx43 mRNA, as to adjust disordered electrogastric rhythm.
9.Long-term Toxicity Study of Compound Zedoary Turmeric Oil Cream for External Use in Rats
Xiaoqian ZHAO ; Ying QIAN ; Shanshan HU ; Xuan LIANG ; Mingjing ZHANG ; Zehui CHEN ; Yingbiao TIAN
China Pharmacy 2016;27(7):882-885
OBJECTIVE:To investigate toxic reaction of Compound zedoary turmeric oil cream in experimental rats with long-term consecutive transdermal administration,and to provide reference for safe use of it in the clinic. METHODS:60 SD rats were randomly divided into blank control (cream matrix) group,Compound zedoary turmeric oil cream intact skin and damaged skin low-dose and high-dose(5%,10%)groups,with 12 rats in each group,half male and half female. All of them were given relevant medicine twice a day. 92 d consecutive medication later,general situation of rats were observed,and body weight,blood routine(WBC,RBC,HGB,LYMPH,etc.)and blood biochemical indicators(AST,ALT,PA,etc.)were all detected;systemati-cal observation of organs,organ coefficient calculation and histopathology examination were carried out. RESULTS:There was no statistical significance in those indicators between Compound zedoary turmeric oil cream groups and blank control group (P>0.05),except hemoglobin decreased in intact skin low-dose group,while hemoglobin decreased,LYMPH and PA increased in dam-aged skin high-dose group(P<0.05). Pathology results showed that Compound zedoary turmeric oil cream had no significant toxici-ty for the main organs. CONCLUSIONS:Compound zedoary turmeric oil cream has no long-term toxicity to experimental rats.
10.Study of the effects of hepatocyte growth factor on inhibit Intimal hyperplasia of the anastomotic stoma after carotid artery bypass grafting
Zhuangjie XING ; Mingjing LIU ; Guoqiang HAO ; Runsheng LI ; Xin ZHENG ; Zhihui DENG ; Hui ZHAO
Journal of Chinese Physician 2012;(z1):18-20
Objective Study of the effects of hepatocyte growth factor on inhibit Intimal hyperplasia of the anastomotic stoma after carotid artery bypass grafting.Methods Thirty-two New Zealand white rabbits were randomly divided into control group and the experimental group.The veins were pretreated with saline solution(control group)only or pretreated with HGF(experimental group ;100ng/ml).The vein grafts were harvested at 14 days,28days after operation,HE Stain and Elastic fibrin Stain,The thickness of Intima and media in the vein grafts,intima-media ratio(I/M) was calculated by computer image analysis system.PCNA Immunohistochemistry was performed.Results The thickness of Intima and media in the vein grafts of control group surpassed experimental group significantly(P <0.01).At 14d I/M in the vein grafts of control group (0.81 ± 0.05) surpassed experimental group (0.47 ± 0.05) (P < 0.01),At 28d I/M in the vein grafts of control group(0.73 ± 0.01)surpassed experimental group (0.65 ± 0.01) (P < 0.01).The vascular smooth muscle cell proliferation in experimental group was significantly lower than that in control group (P < 0.01).Conclusion Treatment of veins grafts with HGF can significantly inhibit intimal hyperplasia in a rabbit carotid artery bypass grafting model.