1.Lentivirus-mediated shRNA interference targeting ADAM17 down-regulates MMP-9 expression in airway epithelial cells via TNF-α/NF-κB signaling
Jianping YAN ; Yaqing LI ; Hui ZHONG ; Chun CHEN ; Chao GU
Chinese Pharmacological Bulletin 2014;(4):559-565
Aim To study the role of TNF-α/NF-κB signaling in matrix metalloproteinase ( MMP)-9 expres-sion induced by lipopolysaccharide ( LPS ) in airway epithelial cells, and to investigate the effects of lenti-virus mediated RNAi targeting a disintegrin and metal-loproteinase 17 ( ADAM17 ) gene on MMP-9 expression induced by LPS. Methods The ADAM17 siRNA ex-pression vector was constructed, and packaged to re-combinant lentivirus in 293T cells. The HBE4-E6/E7 cells were pretreated for 30 min by NF-κB inhibitor ( PDTC) and a recombinant human TNFR p75-Fc fu-sion protein ( Etanercept) , or infected by the recombi-nant lentivirus for 72 h, and then stimulated for 24 h by LPS or TNF-α. The release of TNF-α was detected by ELISA. The mRNA and protein levels of MMP-9 were analyzed respectively by RT-PCR and Western blot. NF-κB activity was detected by electrophoretic mobility shift assay. Results LPS and TNF-α signifi-cantly increased MMP-9 mRNA and protein expressions and the activation of NF-κB in HBE4-E6/E7 cells ( P<0. 05 ) . Etanercept and PDTC significantly inhibited MMP-9 expression and the activation of NF-κB induced by LPS ( P<0. 05 ) . Lentivirus mediated RNAi targe-ting ADAM17 significantly decreased TNF-α produc-tion, inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by LPS in HBE4-E6/E7 cells ( P <0. 05 ) . Lentivirus mediated RNAi targeting ADAM17 did not inhibit MMP-9 mRNA and protein expressions and the activation of NF-κB in-duced by TNF-α ( P>0. 05 ) . And PDTC significantly inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by TNF-α ( P <0. 05 ) . Conclusions TNF-α/NF-κB signaling partic-ipates in the regulation of MMP-9 expression induced by LPS in airway epithelial cells, and lentivirus-media-ted RNAi targeting ADAM17 plays an important role in that signaling pathway upstream by regulating TNF-αrelease.
2.Application of quantitative fingerprint to amino acids composition analysis of Xingnao Tongluo injection.
Jun-Hua HU ; Xia LIN ; Yan ZHONG ; Hui LI ; Jian-Xiong WU ; Jia-Chun LI ; Zhen-Zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2014;39(10):1811-1815
OBJECTIVETo control the quality of the product, quantitative fingerprint was used to evaluate the composition of the amino acids in the Xingnao Tongluo injection.
METHODThe method of the quantitative fingerprint to the amino acids composition was established through AccQ Tag precolumn derivatization. The quality was evaluated by the quantitative test of the amino acids and the similarity in ten batches.
RESULTThe Xingnao Tongluo injection contained 12 amino acids and the contents of these amino acids were stable. All the ten batches of the samples had similarity of more than 0.90.
CONCLUSIONThe method was accurate, feasible and could be a simple and effective way to evaluate the quality of the traditional Chinese medicine.
Amino Acids ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control
3.Effect of sodium houttuyfonate in enhancing imipenem's activity against Pseudomonas aeruginosa biofilms.
Hui-Juan CHENG ; Min YANG ; Chang-Zhong WANG ; Chang-Feng ZHANG ; Chun-Yun CHU
China Journal of Chinese Materia Medica 2014;39(2):296-299
OBJECTIVETo investigate the resistant effect of houttuyfonate sodium (SH) combined with imipenem (IMP) against Pseudomonas aeruginosa (Pa) biofilms.
METHODThe two-fold dilution method was used to examine the minimum inhibitory concentration (MIC) of the tested drug. The crystal violet staining was applied to detect the effect of the combination of 1/2MIC, 1MIC, 2MIC of SH, single IMP, 1/2MIC of SH and IMP of various concentrations on the clearance rate of adherent bacteria, growth of biofilms and alginate production. Fluorescein diacetate (FDA)-propidium iodide (PI) doubling staining assay was employed to observe the bacterial viability and morphological changes after membrane dispersion of each drug group.
RESULTSodium houttuyfonate could enhance the effect of IMP against pseudomonas aeruginosa biofilms. Particularly, the combination group with the concentration of 2MIC showed the highest effect, with P < 0.001 compared with the negative control group. The above results were proved by the bacterial viability and biofilm morphology under fluorescence microscope.
CONCLUSIONAfter being combined with imipenem, sodium houttuyfonate shows a higher effect against biofilms. It is expected that the combination of the two drugs could improve the clinical efficacy of associated infections.
Alkanes ; pharmacology ; Biofilms ; drug effects ; growth & development ; Drug Synergism ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Microbial Viability ; drug effects ; Pseudomonas aeruginosa ; drug effects ; physiology ; Sulfites ; pharmacology
4.A survey of iodine nutrition status of Tianjin residents between 2002 to 2011 and analysis of suitable iodine content in salt
Chang-chun, HOU ; Zhong-hui, LIU ; Shu-qing, HAN ; Qiang, ZENG ; Yang, WANG
Chinese Journal of Endemiology 2013;32(5):533-536
Objective To investigate the consumption status of iodized salt and iodine nutrition status of Tianjin residents,and to provide a scientific basis for iodine supplementation.Methods Sampling methods:① Salt iodine:According to The National Project of Surveillance on IDD,the iodine in salt samples from 18 Tianjin districts (counties) was tested between 2002 to 2011.②Iodine nutritional status of children:Investigation of iodine nutritional status of children was conducted four times in 2002,2005,2009 and 2011.In 2002 and 2005,two primary schools were selected in each district.By age,gender parity principle,40 subjects aged from 8 to 10 in each school were randomly selected to perform thyroid examination and 20 of them were selected to collect urine samples for determination of urinary iodine.In 2009,according to their sub-area positions in the north,the south,the east,the west and the center of each district,5 primary schools were selected in each town (if there were less than five towns in the district,all towns had been selected).Twenty subjects aged from 8 to 10 in each school were selected to collect urine samples for determination of urinary iodine.In 2011,probability sampling method (PPS) was used to select 30 primary schools,and then 40 children aged from 8 to 10 were randomly selected in each school to examine thyroids.At the same time,urine samples from 12 children of the 40 selected children were tested.③Iodine nutritional status of women of childbearing age:In 2007,2008 and 2010,150,50 and 60 women of childbearing age were selected in Hangu District,urine samples of them were collected for determination of urinary iodine.④Iodine nutritional status of pregnant and lactating women:In 2011,3 towns around each primary school were selected.Five pregnant and five lactating women were selected in each town,urine samples of them were collected for determination of urinary iodine.Test methods:①Salt iodine was tested by direct titration,while Sichuan salt and other reinforced edible salt by arbitration determination based on the General Test Method in Salt Industry-Determination of Iodide Ion (GB/T 13025.7-1999).②Thyroid was tested by B-type-ultrasound and judged according to Diagnostic Criterion of Endemic Goiter (WS 276-2007).③Urinary iodine was tested by the Method for Determination of Iodine in Urine by As3+-Ce4+ Catalytic Spectrophotometry (WS/T 107-2006).Results From 2002 to 2011,the consumption rate of qualified iodized salt,the rate of qualified iodized salt,the coverage rate of iodized salt,the rate of non-iodinated salt was 92.7%(43 489/46 926),97.4%(43 489/44 694),95.1% (44 694/46 926) and 4.8%(2273/46 926),respectively.The median salt iodine was in the range of 29.2-36.7 mg/kg.Children's urinary iodine was monitored 4 times,the median urinary iodine was 228.0,221.5,191.8; and 194.7 μg/L,respectively.Children goiter rates were 2.1%(27/1258),1.6%(19/1186) and 2.1%(26/1219) of the 3 times monitored.The median urinary iodine in pregnant and lactating women was 145.2 and 136.0 μg/L.The median urinary iodine in women of childbearing age was 130.7,196.1 and 229.5 μg/L,which increased with the increase of coverage of iodized salt.Conclusions The iodine nutrition of Tianjin residents,women of childbearing age and lactating women are at appropriate level.The iodine nutrition of pregnant women is lower than appropriate level.Recommended salt iodine level in our city is 30 mg/kg,or 25 mg/kg for ordinary residents,and 30 mg/kg for pregnant women.
5.Investigation on the prevalent condition of adult osteofluorosis in the endemic fluorosis areas of Tianjin in 2008
Chang-chun, HOU ; Shu-qing, HAN ; Zhong-hui, LIU ; Hong-lian, LIU
Chinese Journal of Endemiology 2010;29(3):322-324
Objective To investigate the prevalence of adult osteofluomsis in the endemic fluomsis areas in Tianjin and to provide scientific foundation for endemic fluorosis.Methods Stratified sampling in 55 villages were selected in 3 areas with slight,moderate and severe fluorosis regions in Tianjin from April to June in 2008.Water fluorine were tested and clinical osteofluorosis examinations were conducted to the population aging 16 and above in the villages.Tweenty villages were selected randomly in the slight,moderate and severe fluorosis regions.X-ray osteofluorosis examination were conducted to patients and suspected patients in these 20 villages.Results The geometric mean fluoride content in the water for the 3 areas were 1.35 mg/L,3.44 mg/L,5.49 mg/L,respectively.The prevalence of osteofluorosis were 36.7%(44/120),20.6%(33/160),39.4%(43/109),respectively.The prevalence of osteofluorosis Was increased gradually(r=0.534,P<0.01)and the symptoms and signs of the disease were more serious(H=84.813,P<0.01).The prevalence of X-ray diagnosis Was increased gradually(r=0.990,P<0.01)and signs of the disease were more severe(H=25.169,P<0.01)with an increase in age.There was no statistical significance of prevalence rate of osteofluorosis between males and females,regardless if it Was a clinical diagnosis(X2=0.343,P>0.05)or an X-ray diagnosis(X2=3.532,P>0.05).Conclusions Adult osteofluorosis to a certain extent is still prevalent in the fluorosis areas in Tianjin.Endemic fluorosis is still rampant.Improving water in fulorosis areas should be mandatory.
6.A comparative study of hemodynamics after cardiac surgery with venous-arterial modified ultrafiltration and aterial-venous modified ultrafiltration in infants
Hui ZHONG ; Xue-Cun LIANG ; Zhang-gen CHEN ; Chun-Ian HE ; Bing JIA ;
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(7):412-415
Objective Evaluate the effects of venous-arterial modified ultrafiltration on hemodynamics compared to arterial-venous in children undergoing cardiopulmonary bypass (CPB) for repair of congenital heart defects. Methods Forty patients underwent MUF randomly divided into two groups,group V-A MUF (n =20) and group A-V MUF (n =20) for 10 min after CPB. They were studied before CPB, after CPB, 10 min after CPB, and 30 min after CPB. Haemodynamic data including heart rate, blood pressure, central venous pressure and hematocrit were recorded. Transoesophaegeal echocardiography determined left ventricular posterior wall thickness in end-systole ( LVPWs) and end-diastole (LVPWd) , end diastolic volume (EDV) , end systolic volume (ESV) and ejection fraction (EF) were measured and compared in two groups. Results Patients in V-A MUF maintained better systolic arterial blood pressure at 10 min and 30 min compared with 0 min values after CPB. A significant decrease in EF were observed in both groups immediately after CPB ( P < 0.05 ). Significant increase in EF was observed at 10 min (60% ) and 30 min (46% ) after CPB compared with 0 min value after bypass in V-A MUF (P <0.001 ). In A-V MUF, no such increase in EF was observed. EF were significantly higher at 10 min and 30 min in V-A MUF as compared with A-V MUF (P < 0. 001). There was also significant improvement in posterior wall thickness in V-A MUF (P <0.05). Haematocrit values were not different in duration of postoperative between two groups. Conclusion Veno-arterial modified ultrafiltration is a safe and effective method of improving hemodynamics in children following cardiac surgery.
7.Research about the expression changes of CNTF and CGRP in the rabbits' model of neuroma-in-continuity
Chun-Hui SONG ; Tong-Yi CHEN ; Feng ZHANG ; Jian ZHANG ; Zhong-Wei CHEN ;
Chinese Journal of Microsurgery 2000;0(03):-
Objective In view of being short of the mammalian model in neuroma-in-continuity,the experiment injured the part of peroneal nerve to the formation of the neuroma-in-continuity and was applied to the foundation of farther research.Methods Twelve New Zeland rabbits were selected as experimental sub- jects randomly.One lateral peroneal nerves of twelve rabbits were resected,the damaged nervous tissues' slice were showed to the typical pathological changes of neuroma by the stain of HE,luxol fast blue after six weeks. As compared with the health sides of six model rabbits,the methods of real-time PCR and Western blot were used to evaluate the expression of CNTF,CGRP mRNA and protein in injured nerves and L_7、S_1 dorsal root ganglions respectively.Results The injured nerve formed the typical pathological changes of neuroma at six weeks.Compared with hea|thg side the expression of CNTF mRNA and protein was down-graded at the lateral of neuroma(P<0.05),and the expression of CGRP mRNA and protein was up-graded(P<0.05).Con- clusion The method of partly injuring the peroneal nerve could effectively set up the model of the neuroma-in- continuity,furthermore,resulted to the expression changes of the CNTF,CGRP mRNA and protein.
8.Clinical research of synchronous colorectal neoplasms.
Hui WANG ; Zhong-Guo ZHANG ; Chun SONG ; Bin WANG
Chinese Journal of Surgery 2006;44(21):1498-1500
OBJECTIVETo investigate the clinical and pathological features of synchronous colorectal carcinomas and explore the specificity of treatment for those patients.
METHODSA retrospective evaluation of 326 patients with primary colorectal cancer were performed. The patients were divided into two groups: single cases (303 patients) and synchronous cases (23 patients, index cancer and second cancer). Clinical characteristics, routine pathological findings in the two groups were compared statistically.
RESULTSThere was no difference in location of the lesions between the single cases and the index lesions of synchronous cases, while the differentiation and Ducke stage of index lesions were better than those in single cases. The secondary cancer of synchronous cases showed better grades and stages than those of single cases. The frequencies of complicating adenoma and multiple polyps in synchronous cases were higher significantly than those in single cases, with the rate of 34.8% and 78.3%, 11.6% and 42.2%, respectively. All malignant lesions were resected radically, and multiple polyps (56.9%) were electro-resected under colonoscopy at 2 approximately 3 months after the carcinomas were radically resected. Of the patients complicated with adenomas, 79.1% (34 cases) were given one-stage local resection or local colectomy and the rest were given second stage electro-resection.
CONCLUSIONSFor the clinical and pathological features of synchronous colorectal carcinomas, it is essential to perform a complete evaluation of all colorectal lesions, establish an individual surgical treatment and follow-up in time.
Adult ; Aged ; Aged, 80 and over ; Biomedical Research ; methods ; Colorectal Neoplasms ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Neoplasms, Multiple Primary ; pathology ; surgery ; Retrospective Studies
9.Study on the Yersinia pestis plasmid along the Qinghai-Tibet Railway
Chun-hua, ZHANG ; Jing-sheng, L(U) ; Bin, ZHAO ; Xian-bin, CONG ; Zhong-hui, WANG ; Shi, ZHANG ; Kui-dong, SHAO
Chinese Journal of Endemiology 2008;27(6):595-597
Objective To detect the Yersinia pestis plasmid and molecular weight along the Qinghai-Tibet Railway.Methods Yersinia pestis plasmids molecular weight detected and analyzed using alkaline lysis,phenol-chloroform extraction of Yersinia pestis plasmid by agarose gel electrophoresis.Results The 18 Yersinia pestis strains of Qinghai-Tibet Railway contained 6×106,45×106,52×106,65×106,92×106plasmid,varing in the range of the 52×106-92×106.Conclusions The Yersinia pestis of Qinghai-Tibet Railway has a standardplasmid graphics,with the biggest Yersinia pestis plasmid changing in a certain regular degree,which providessignificance in the study of plague natural foci of the spatial structure and the genetic.characteristics of Yersiniapestis.
10.Fluorescence labeling for human bone marrow mesenchymal stem cells with PKH26
Xing-Zhong WANG ; Wen-Rong XU ; Wei ZHU ; Huan YANG ; Chun QIAO ; Hui QIAN ; Jia-Bo HU ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To establish a method of labeling human mesenchymal stem cells (MSCs) with PKH26 in vitro.Methods MSCs were cultured and labeled with PKH26 according to the manufacturer's instruction.The growth,fluorescence intensity and serial subcuhivation of labeled MSCs were analyzed with the confocal laser microscope and the flow cytometry.The biological characteristics of labeled MSCs were investigated by RT-PCR.Results The labeled MSCs appeared red fluorescence and the labeling rate was 100 percent.During serial subcuhivation of labeled MSC from passage 1 to passage 7,the fluorescence intensity and the labeling rate of MSCs were gradually decreased.The biological features such as morphology,growth,expression level of nucleostemin and GAPDH gene and capability of differentiation into osteoblast in vitro were not affected by labeling.Conclusion Labeling the human MSCs with PKH26 is an effective and practical method,which can be used as an important tool in the study on the homing, plasticity and transplantation of MSCs.