2.Change of matrix metalloproteinase-9 in rats with multiple organ dysfunction syndrome
Min YU ; Guanlan LI ; Xianzhe LIU ; Min QIAN ; Xinli ZHANG ; Yuqi WU ; En LV ; Changyun GUO
Chinese Journal of Emergency Medicine 2009;18(2):165-169
Objective To investigate the changes in the serum MMP-9 (matrix metalloproteinase-9) and the expressions of MMP-9 in lung, kidney and intestine in rats with multiple organ dysfunction syndrome (MODS) and confirm extracellular matrix injuries being the mechanism in MODS in order to propose a novel theoretical basis for cfinical treatment of MODS. Method Forty wister rats were randomly divided into two groups: control group (n=8) and MODS model group (n=32). The rats of model group were further divided into four subgroups ac-cordingto the time elapsed after modelling: 12 h (n=8), 24 h(n=8) ,48 h(n=8) and 72 h (n=8), and were modelled by celiac injection of mixed liquid of zymosan-paraffin (4 mL/100 g) after blood loss (1mL/100 g) by extirpating their left eyes. Blood,lung, kidney and intestine were sampled 12,24,48 and 72 hours after models were established. The histological changes in the lung, kidney and intestine of the rats were observed by light mi-croscope. The serum MMP-9 were measured by enzyme-linked immunosorbent assay (ELISA). The immunohisto-chemistry was used to observe the expression of MMP-9 in lung,kidney and intestine during different phases of MODS. The data were processed by one-way ANOVA and Bivariate analysis. Results Compared with control group, the organs were injured by congestion, edema and inflammatory cells infiltration to a certain extent in model groups. The serum MMP-9 increased markedly 12 hours after modelling (P<0.01 ) and peaked 48 hours later. The expressions of MMP-9 in lung, kidney and small intestine significantly increased from 12 h to 72 h after mod-elling (P<0.01 or 0.05). Conclusions The MMP-9 increased both in serum and tissue are closely associated with the pathological process of MODS. The mechanism of organ damage probably attributes to the damage of extra-celluar matrix and tissue construction.
3.Preliminary evaluation on the detection of serum level of antibody to Bacillus anthracis by enzyme-linked immunosorbent assay using crude antigen
Jian-chun, WEI ; Hui-juan, ZHANG ; Feng-qin, MA ; En-min, ZHANG ; Dong-zheng, YU
Chinese Journal of Endemiology 2008;27(3):329-331
Objective To evaluate the method of detecting antibodies to Bacillus anthracis by enzymelinked immunosorbent assay(ELISA)using crude antigen.Methods The anti-Bacillus anthracis antibody levels in sera of 42 healthy people and 42 patients were detected by indirect ELISA.Standard curve was plotted using the data from positive controls,based on which the relative content of each serum was calculated and compared with the result of rLF.Results The median of antibody's relative content in patient group and healthy people group are 1.19 and 0.24,the differences being statistically significant(uc=7.643,P<0.05).The result of crude antigen is in concordance with rLF(but not parallel absolutely).Conclusions Crude antigen can distinguish most of patients with healthy population effectively.The results suggested that crude antigen is applicable in anti-Bacillus anthracis antibody surveillance.
4.Cloning and expression of the specific genes from Yersinia pestis and analysis of their antigenicity
Ying, LIANG ; Rong, HAI ; Dong-zheng, YU ; Jian-chun, WEI ; Zhi-kai, ZHANG ; En-min, ZHANG ; Jian-hua, ZHANG
Chinese Journal of Endemiology 2008;27(5):503-507
Objective To clone and express specific genes (YP01089,psi.ymt) from Yersinia pestis in Escherichia Coli(E.coli)and to analyze the antigenicity of these recombinant proteins.Methods The target genes were amplified by polymerose chmn reaction(PCR).The amplified products were ligated with pET-30a(+) vector after purification and cut by two different restriction enzymes,then these recombinant plasmids were transfefred into the host cells of E.coli BL21(DE3) strain.The target genes were successfully expressed following induction with Isopropyithio-β-D-galactoside(IPTG),and the target proteins were purified by the method of affinity chromatography.Sodium dodecyl sulfate-polyaerylamide gel eleetrophoresis(SDS-PAGE)and Westem blot were used to detect the expressed recombinant protein.Results Three recombinant plasmids were finally constructed. rYP01089,rPst and rYmt were expressed stably and effectively in E.coli thmngh optimizing the induction condition. The Western blot analysis indicated that rPst was capable of binding with positive sernm of phgue.The purity of rest was up to 95%in this stuay.Conclusions This work indicates that the genes of Yersinia pestis are able to be efficiently expressed in the prokaryotie protein expression system.The immune characteristic of rPst is sensitive and specific,80 this study has settled a foundation for developing a new type diagnostic reagent of plague.
5.Changing trends of the expression of TIMP-4 in mouse ovary during pregnant and postpartum period.
Shu-Min BU ; Zeng HU ; Fan ZHANG ; En-Kui DUAN
Chinese Journal of Applied Physiology 2008;24(2):248-252
AIMThe changes of tissue inhibitor of metalloproteinase-4 (TIMP-4) expression in mouse ovary during pregnant and postpartum period were studied to investigate the role of TIMP-4 in corpus luteum (CL).
METHODSRT-PCR was used to deter mine the change of TIMP-4 mRNA and indirect immunofluorescence was used to observe the change of TIMP-4 protein. The expression of TIMP-4 mRNA was observed in various periods throughout the stage of pregnancy and postpartum day 1.
RESULTSThe expression of TIMP-4 was gradually enhanced from day 1 to day 8, reached a maximal expression at day 8, while decreased at day 11 and to the lowest level at postpartum day 1. Indirect immunofluorescence results further indicated that TIMP-4 protein was localized to CL and theca-intera cells in various periods throughout the pregnancy and postpartum day 1. In addition, the change pattern of TIMP-4 protein agreed with that of the TIMP-4 mRNA in pregnancy CL.
CONCLUSIONThe expression of TIMP-4 in mouse ovary during pregnancy and postpartum is in spatio-temporal pattern and it may be involved in the formation and function maintain of CL during pregnancy in mice.
Animals ; Female ; Mice ; Mice, Inbred Strains ; Ovary ; metabolism ; Postpartum Period ; Pregnancy ; RNA, Messenger ; genetics ; Tissue Inhibitor of Metalloproteinases ; metabolism
6.Clinical study on treatment of severe acute respiratory syndrome by integrative Chinese and Western medicine.
Ai-min REN ; Shu-wen ZHANG ; Bao-en WANG
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(2):112-114
OBJECTIVETo evaluate the effect of integrative Chinese and western medicine (ICWM) in treating severe a cute respiratory syndrome (SARS).
METHODSSixty SARS patients were diagnosed and observed according to the universal standard, and divided into the ICWM group (n = 31, treated with ICWM) and the control group (n = 29, treated by conventional western medicine alone).
RESULTSICWM showed better effect than that of western medicine alone in improving clinical symptoms, promoting the absorption of inflammation in lung, increased oxygen saturation (P < 0.01) and decreased the dosage of corticoid used (P < 0.05).
CONCLUSIONThe effect of ICWM is better than that of simple western medicine in treating SARS.
Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Methylprednisolone ; therapeutic use ; Middle Aged ; Phytotherapy ; Ribavirin ; therapeutic use ; Severe Acute Respiratory Syndrome ; drug therapy ; immunology ; Single-Blind Method
7.Isolation of antifungal compound from Paeonia suffruticosa and its antifungal mechanism.
Yong ZHAO ; Bao-en WANG ; Shu-wen ZHANG ; Shu-min YANG ; Hong WANG ; Ai-min REN ; En-tong YI
Chinese journal of integrative medicine 2015;21(3):211-216
OBJECTIVETo isolate antifungal compound from Paeonia suffruticosa, and to find the antifungal mechanisms by observing the ultrastructural modifications of yeasts in growth phase produced by 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (PGG).
METHODSPeony (Paeonia suffruticosa) root bark (PRB) was separated by solvent extraction and purified by high performance liquid chromatography (HPLC) method using analytical and preparative reversed phase C18 column on the basis of bio-assay method. In order to investigate the antifungal mechanism of PGG, Yeasts were submitted to different concentrations [3 × minimum inhibition concentration (MIC), 0.3 × MIC] for 1 h under constant stirring at 30 °C, and transmission electron microscopy was performed.
RESULTSBased on the antifungal activity of PRB on Candida glabrata CBS138, the antifungal compound were isolated in ethyl acetate layer of PRB and identified as PGG by mass spectrometry, 1H nuclear magnetic resonance (NMR) analyses, with molecular weight of 940 and molecular formular as C41H32O26. Transmission electron microscopy showed that PGG degraded the cell wall envelope.
CONCLUSIONThe results suggest that PGG may be responsible for the antifungal activity of PRB by disrupting the structure of cell wall directly.
Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Candida ; drug effects ; ultrastructure ; Chromatography, High Pressure Liquid ; Hydrolyzable Tannins ; chemistry ; isolation & purification ; pharmacology ; Mass Spectrometry ; Microbial Sensitivity Tests ; Paeonia ; chemistry ; Plant Bark ; chemistry ; Plant Extracts ; isolation & purification ; pharmacology ; Plant Roots ; chemistry ; Proton Magnetic Resonance Spectroscopy
8.Experience in the removal of difficult and high risk tracheobronchial foreign body by bronchoscopy.
En-ming XU ; Zhong-qiang XU ; Zhi-nan WANG ; Yan WANG ; Ping CHEN ; Ya-min ZHANG ; Fei XIA ; Yan ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(12):982-986
OBJECTIVETo explore the best methods and skill for the removal of difficult and high risk tracheobronchial foreign body under bronchoscope.
METHODSA retrospective review was performed between August 1995 to August 2012. There were 4217 children with tracheobronchial foreign body, among them, 272 were diagnosed as high-risk, highly difficult tracheobronchial foreign bodies confirmed by clinical manifestations, foreign body type and bronchoscopy.
RESULTSIn 271 children, the tracheobronchial foreign body was removed under bronchoscope, the success rate was 99.6%; only one child with a pen cap blocking the left lower lobe bronchus was transferred to the department of thoracic surgery, and the foreign body was finally removed by thoracotomy. Eighty-five children (among them, 82 children were under 1 year of age) had II-II degree laryngeal obstruction, the emergency surgery was performed to remove the foreign body and to relieve the laryngeal obstruction. Twenty-six children had lung infection and 27 children had failed foreign body removal surgery before, in all these children, the foreign body was removed after infection control. There were 17 children with the pen cap as the tracheobronchial foreign body, direct removal was successful in 12 children with the history less than two weeks; in 4 children, the foreign body was removed after 0.1% epinephrine saline flush, and 1 case with the homemade bronchial foreign body hook remove. There were 26 children with the whistle as the foreign body, and 32 children had large and sharp foreign bodies. In these cases, the foreign bodies were removed together with the bronchoscope. Forty-two children had multiple or fragile foreign bodies, and 16 children had subsegmental bronchial foreign bodies. In these cases, the foreign bodies were removed with forceps under direct vision and intraoperative bronchial lavage.In This series, 129 children received intraoperative bronchial lavage, among them, 127 children showed normal X-ray changes one week after operation. Two children with a history of more than 1 month complicated with pulmonary consolidation. After bronchial lavage, pneumothorax and subcutaneous emphysema occurred, which recovered after treatment. No glottic edema, asphyxia, and other complications were found, the complication rate of surgery was 0.7%.
CONCLUSIONFor the removal of highly difficult and high risk tracheobronchial foreign bodies, preoperative analysis and discussion should be sufficient, appropriate surgical skill and surgical instruments may improve the success rate of the surgery and prevent the operation complications.
Adolescent ; Bronchi ; Bronchoscopy ; methods ; Child ; Child, Preschool ; Female ; Foreign Bodies ; surgery ; Humans ; Infant ; Male ; Retrospective Studies ; Trachea
9.Study on the internal control on polymerase chain reaction in Yersinia pestis detection.
Zhi-kai ZHANG ; Rong HAI ; En-min ZHANG ; Dong-zheng YU
Chinese Journal of Epidemiology 2005;26(1):36-38
OBJECTIVEFor the detection of Yersinia pestis by polymerase chain reaction (PCR), internal control (IC) is required in order to prevent false negative results that might be caused by PCR inhibitors.
METHODSF1 antigen was amplified by PCR with primer F1 and the PCR product of primer F1 were cloned with TOPO TA cloning Kit. The plasmid of positive clone was then digested with HpaI. The digested plasmid and the PCR products of 16S rRNA were ligated with T(4) DNA ligase before the ligated products were transformed. Isolate plasmid DNA on positive clone and its concentration were measured. Plasmid DNA on different concentration by PCR amplification with primer F1 was analyzed and the standard concentration of IC was determined.
RESULTSConstructing an IC by inserting a 16S rRNA amplicon to the original target DNA between the two primer F1 sites, the size was longer than the target DNA. The standard concentration of IC was determined.
CONCLUSIONAn optimal IC concentration to increase the reliability of the PCR assays might be used to prevent false negative results and appeared to be useful for detection of Yersinia pestis.
Bacterial Proteins ; analysis ; DNA, Bacterial ; analysis ; False Negative Reactions ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; Sensitivity and Specificity ; Yersinia pestis ; isolation & purification
10.Study on the relation between the absence of one IS100 in 102 kb pgm locus of Yersinia pestis and the stability of pigmentation phenotype.
En-min ZHANG ; Rong HAI ; Zhi-kai ZHANG ; Dong-zheng YU
Chinese Journal of Epidemiology 2004;25(10):886-889
OBJECTIVETo study the relation between the absence of one IS100 in the 102 kb pgm locus of Yersinia pestis and the stability of pigmentation phenotype (pgm(+)).
METHODSWe amplified the segment including IS100 in 102 kb pgm locus of Yersinia pestis that isolated from all ecotypes in China by polymerase chain reaction (PCR). There were 171 strains isolated from 18 ecotypes in this study. One strain was chosen to be cloned and sequenced.
RESULTSBesides the type of Microtus brandti, the types of East-North Tianshan, A and B of West-North Tianshan, Microtus Qinghai had one band with about 2560 bp. These strains lost one IS100 in 102 kb pgm locus of Yersinia pestis. Their pgm(+) phenotype was stable. Some strains of ecotypes from Qilian Mountain, Qinghai-Tibet Plateau, Gangdisi Mountain, West Yunnan Mountain had no bands in the PCR products. Negative strains would lose the whole 102 kb pgm locus. The others had one band with 4492 bp. These strains had two IS100 which flanked the 102 kb pgm locus but the pgm(+) phenotype was unstable.
CONCLUSIONYersinia pestis which had only one IS100 would flank the 102 kb pgm locus and had stable pgm(+) phenotype while the Yersinia pestis that having two IS100 flanked the 102 kb pgm locus would have unstable pgm(+) phenotype.
Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; DNA, Bacterial ; genetics ; Genetic Variation ; Genomic Instability ; Phenotype ; Pigmentation ; genetics ; Yersinia pestis ; genetics