1.Process concerning the periprosthetic infection after total hip replacement
Chinese Journal of Tissue Engineering Research 2011;15(43):8118-8123
BACKGROUND: Periprosthetic infection after total hip replacement usually results in surgery failure and needs a second operation.OBJECTIVE: To explore the pathogenesis, diagnosis and treatment of periprosthetic infection after total hip replacement by-reviewing and summarizing articles published in recent years.METHODS: A computed-based online search of Pubmed database was performed by using the key words of total hip arthroplasty, infection for manuscripts published from January 1990 to December 2010 and of those in SpringerLink database between January 1,1970 and December 31, 2010. A total of 2 109 manuscripts were retrieved. Moreover, related book or manuscripts that published by high-impact journals were included. Totally, 29 manuscripts were included. RESULTS AND CONCLUSION: The formation of biofilms on the surface of prosthesis is the main cause for hardly eradicated. Resistant bacteria and polymicrobial infection seems to be an increasing tendency. A correct diagnosis as soon as possible is very important to prognosis. However, without a gold-standard way, each mean has advantages and shortages, and comprehensive considerations are necessary. lnterleukin-6 seems a good choice for its inexpensive, non-invasive and a high sensitivity and specificity, which has aroused increasing attention. The antibiotics only, debridement with retention, one-stage replacement, two-stage replacement, joint arthrodesis, even amputation, are used to treat infection after total hip replacement. Prophylactic antibiotics are important to prevent infection. Antibiotic-loaded acrylic cement seems to be reliable and accept for more and more patients. However, each option must be selected according to the presence of infection individually.
2.The detection of telomerase activity and expression of human telomerase reverse transcriptase mRNA
Yirong LI ; Jianmin WU ; Lihua HU
Chinese Journal of Immunology 2000;0(11):-
Objective:To set up a real-time fluorescent RT-PCR and to quantitate the expression levels of hTERT mRNA in peripheral blood mononuclear cells from patients with AML and to observe the correlation between the expression of hTERT mRNA and occurrence and relapse of AML and to probe into the correlation between the expression of hTERT mRNA and telomerase activity.Methods:Real time fluorescent RT-PCR and Lightcycler PCR system were used to quantitate expression levels of hTERT mRNA.PCR-ELISA was used to quantitate telomerase activity.Results:①N_(hTERT) from AML at initial presention,AML at relapse,AML at complete remission and health examinee was 299.2?292.8,550.1?441.3,14.0?9.2 and 12.3?6.7 respectively and the expression levels of hTERT mRNA from AML at initial presention and AML at relapse elevated signficantly comparing to that from AML at complete remission and health examinee,moreover the expression levels from AML at relapse was higher than that from AML at initial presention significantly.②The telomerase activity from AML at initial presention,AML at relapse,AML at complete remission and health examinee was 32.8%?24.3%,48.6%?31.4%,7.4%?5.1% and 7.6%?3.6% respectively and the telomerase activity from AML at initial presention and AML at relapse elevated signficantly comparing to that from AML at complete remission and health examinee,moreover telomerase activity from AML at relapse was higher than that from AML at initial presention significantly.③There was a strong correlation between telomerase activity and the expression of hTERT mRNA and the correlation coeffecients was 0.78.Conclusion:The up-regulation of the expression levels of hTERT mRNA and telomerase activity are one of important factors during occurrence and relapse of AML,moreover there is a strong correlation between telomerase activity and the expression of hTERT mRNA.
3.Real-time Quantitative Assay of Insertion Sequence 6110 DNA of Mycobacterium Tuberculosis and Its Value for Diagnosis of Tuberculosis
Xiang HUANG ; Yirong LI ; Zhengxue WU
Journal of Chinese Physician 2001;0(06):-
Objective To set up a real-time quantitative assay method of insertion sequence 6110 DNA of mycobacterium tuberculosis, and explore its value for diagnosing tuberculosis. Methods Real-time quantitative assay of insertion sequence 6110 DNA of mycobacterium tuberculosis was performed with Taqman technique and Lightcycler quantitative PCR system. Results 213 clinical samples of different types were detected, 32 cases were positive, and the positive rate was 15 02%. The scope of quantitative results of positive samples was 3 1~7 2?10 6 copies/ml, and the sensitivity and specificity for diagnosis of tuberculosis were 82 76% and 95 65% respectively Conclusion Real-time quantitative assay of insertion sequence 6110 DNA of mycobacterium tuberculosis is a rapid and effective method for diagnosis of tuberculosis.
4.Upregulation of phagocytic clearance of apoptotic cells by autoimmune regulator.
Liang, SHI ; Lihua, HU ; Yirong, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(2):145-8
To investigate the effect of autoimmune regulator (AIRE) on phagocytic clearance of apoptotic cells, a recombinant expression vector containing full-length human AIRE cDNA was transfected into 16HBE cells. After incubation with transfected 16HBE cells, engulfment of apoptotic HL-60 cells induced by camptothecin was detected by myeloperoxidase (MPO) staining. The change in the expression of Rac 1 in transfected 16HBE cells was determined by RT-PCR and Western blotting. The results showed that the phagocytosis percentage of the experimental group, the mock transfection group and the negative control group (non-apoptotic cells) was (25.50+/-3.67)%, (6.25+/-1.58)% and (1.0+/-0.67)%, respectively. Moreover, the expressions of Rac 1 mRNA and protein were up-regulated in AIRE-transfected 16HBE cells, suggesting that AIRE may function as a regulator in the phagocytic clearance of apoptotic cells by promoting the expression of Rac 1.
5.RNA Interference and its Applications in Cancer Therapy
Fuguo WU ; Yirong CHEN ; Xuechun WANG ; Li CHENG
Journal of International Oncology 2007;34(11):862-866
RNA interference(RNAi), a highly conserved process of post-transcriptional gene silencing, can be induced by small interfering double-stranded RNA that mediate sequence-specific mRNA degradation. In the past several years, RNAi has been widely used as both an experimental tool to study mammalian gene function and a potential therapeutic approach to treat human diseases. In addition, some new proteins which involve in RNAi pathway have been characterized in mammalian cells. Here, we summarize the various molecules in RNAi, mechanism of action, and the current therapeutic applications in cancers.
6.The expressions of endothelin-1 and endothelin A/B receptors mRNA in tissue of benign prostatic hyperplasia treated by daily low-dose sildenafil
Hongde CHEN ; Xueting YE ; Yirong ZHANG ; Zhiliang WENG ; Chendi LI
Chinese Journal of Geriatrics 2012;31(6):500-502
Objective To observe the mRNA expressions of endothelin-1(ET-1) and endothelin A/B receptors (ETA/B) in tissue of benign prostatic hyperplasia treated by daily low-dose sildenafil.Methods A total of 32 patients with benign prostatic hyperplasia were randomly divided into two groups:treatment(25 mg sildenafi for 12 weeks,n=16) and control (no drug,n=16) groups.Immunohistochemical staining,ELISA and RT-PCR were used to detect the expression levels of ET-1 protein and ET A/B mRNA,respectively.Results The expressions of ET-1 protein and ET A/B mRNA in prostatic tissue were significantly lower in treatment group than in control group[(53.31±18.56) ng/kg vs.(83.34±31.38) ng/kg,0.356±0.056 vs.0.624±0.083,0.721±0.083 vs.0.933±0.905,t=-3.295,10.715,6.937,all P<0.001].Conclusions Daily low-dose sildenafil can reduce the expressions of ET-1 and ET A/B receptors mRNA in benign prostatic hyperplasia.
7.Screening the mimotopes of blood group A antigen by phage display technology
Zhaoming TANG ; Lihua HU ; Yirong LI ; Tianpen CUI ; Juan XIONG
Chinese Journal of Laboratory Medicine 2008;31(4):440-445
Objective To explore the peptides that can mimic the blood type A antigen and evaluate the anti-A antibody detection value of these peptides.Methods The anti-A monoclonal antibodv (NaM87-1F6)was used to panning the phage clones from a phage display 12-mer peptide library.Positive clones were identified by phage ELISA,phage mieropanning methods.Phage DNA Was sequenced and the corresponding peptide sequences were deduced.Agglutination inhibition test WaS performed to assess the ability of phage clones to inhibit the binding between the type A red blood cell and the anti-A antibody. ABO-ELISA based on the selected peptides was compared with classical haemagglutination test jn the detection of senlm anti-A antibody.Results Seven positive clones were chosen after panning,phage ELISA and phage micropanning.Six clones displayed peptide EYWYCGMNRTGC(C5),the other one displayed peptide QIWYERTLPFTF(C17).The phages displaying the selected peptides could specifically inhibit agglutination of type A red blood cells(RBCs)by anti-A antibodies.In the ABO-ELISA based on C5 and C17,the receiver operating characteristic(ROC)Curve showed that area under curve(AUC)were 0.889 (P=0.000),0.75l(P=0.000)respectively.The Spearman correlation Coeffieient between the ABO-EliSA value and the antibody titer derived from haemagglutination assay were 0.743(P<0.01),0.664(P<0.01)respectively.As for C5,0.300 was the best cut-off for ABO-ELISA with 82.2% sensitivity and 83.3% specificity.As for C17,the sensitivity and specificity of ABO-ELISA was 68.9% and 63.3% respectively when the cut-off value was 0.250.Conclusions The peptides EYWYCGMNRTGC and QIWYERTLPFTF can mimic the blood type A antigenic epitope.ABO-ELISA based on these peptides has the potential for the detection of anti-A antibody.
8.The comparison among extraction methods of exosomes from T cells of rats
Zhen SUN ; Chibing HUANG ; Yajun SONG ; Yirong CHEN ; Chuangui LI
Chongqing Medicine 2013;(29):3512-3514,3517
Objective The purification methods of the exosomes derived form T cells were established in order to get high quan-tity exosomes .Methods Exosomes from T cells culture supernatants were purified by ExoQuick Precipitation ,ultrafiltration and sucrose gradient centrifugation ,differential ultracentrifugation ,and confirmed via using transmission electron microscopy .The pro-tein expression of the exosomes were analyzed by SDS-PAGE electrophoresis .Western blotting was used to test the expression of IL-2 .Results The protein concentration of the exosomes purified through ExoQuick Precipitation ,ultrafiltration and sucrose gradi-ent centrifugation were higher than through differential ultracentrifugation (P<0 .05) .SDS-PAGE displayed the difference among the exosome purified by three methods .Three kinds of exosomes all expressed IL-2 .Conclusion ExoQuick Precipitation ,ultrafiltra-tion and sucrose gradient centrifugation technique can obtain high purity and complete exosome sample .
9.Improvement of Quality Standard of Reduqing Oral Liquid
Sheng WANG ; Li CHEN ; Yirong FENG ; Min CAI
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(3):75-77
Objective To improve the quality standard of Reduqing Oral Liquid. Methods TLC was used to identify the featured spots of Glycyrrhizae Radix et Rhizoma, Belamcandae Rhizoma and Scutellariae Radix;HPLC method was usded to detect the contents of Baicalin in Reduqing Oral Liquid. Results TLC could identify the featured spots of Glycyrrhizae Radix et Rhizoma, Belamcandae Rhizoma and Scutellariae Radix; Baicalin was detected in range of 53.60–536.00 μg/mL with good linear relationship (r=0.999 99), the average recovery rate was 99.85%, and RSD was 0.63%(n=6). Conclusion The method is simple, accurate and reliable, which can be applied to the quality control of Reduqing Oral Liquid.
10.Detection of telomerase activity and the expression of telomerase subunits in the patients with acute myelogenous leukaemia.
Yirong, LI ; Jianming, WU ; Lin, WANG ; Fenghua, CHEN ; Lihua, HU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(1):48-51
Telomerase activity and the expression of telomerase subunits (for example, telomerase reverse transcriptase and telomerase associated protein 1 and telomerase RNA component) of peripheral white blood cells were detected in the patients with acute myelogenous leukaemia (AML) and the correlation between telomerase activity and the expression of telomerase subunits was observed. In 94 peripheral white blood cells from 18 healthy volunteers and 76 patients with AML, including 31 AML at initial presentation, 24 at relapse and 21 at complete remission, the telomerase activity and telomerase subunits mRNA or RNA were detected by PCR-ELISA and RT-PCR respectively. The results showed that the positive rate of telomerase from patients with AML at initial presentation, at relapse and at complete remission was 74.1%, 79.2% and 4.8% respectively. The positive rate of telomerase reverse transcriptase mRNA from healthy volunteers, AML at initial presentation, AML at relapse and AML at complete remission was 5.6%, 80.6%, 83.3% and 9.5% respectively. The positive rate of telomerase associated protein 1 mRNA and telomerase RNA component in all samples were 100%. It was suggested that the up-regulation of telomerase activity and the expression of telomerase reverse transcriptase is correlated closely with the occurrence and relapse of AML, so telomerase activity and the expression of telomerase reverse transcriptase may be used to estimate the curative effect and predict relapse of AML. Moreover, the up-regulation of telomerase activity is correlated with the expression of telomerase reverse transcriptase significantly.
Carrier Proteins/*metabolism
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DNA-Binding Proteins
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Leukemia, Myelocytic, Acute/*enzymology
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RNA/metabolism
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RNA, Messenger/metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Telomerase/*metabolism