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.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.
4.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.
5.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.
6.The expression and meaning of bcl-xL in superficial bladder tumor
Bing CAI ; Wei WU ; Yeping LI ; Yirong ZHANG
Chinese Journal of Postgraduates of Medicine 2010;33(5):15-17
Objective To investigate the relations of the expression of bcl-xL in superficial bladder tumor and prognosis. Method The expression of bcl-xL was detected by envision system in 80 cases of superficial bladder tumor. Results In patients with high expression of bcl-xL, the recurrence rate was higher than that of normal expression [71.4%(30/42) vs 50.0%(19/38) ,P < 0.05], and the recurrence of time as early as normal expression [(16.0 ± 1.2) months vs (36.0 ± 4.5) months](P < 0.05). Conclusion The expression of bcl-xL could effectively predict the prognosis of patients with bladder tumor, those who with high expression of-bcl-xL have bad prognosis.
7.Study on Fap1 glycosylation and maturation regulating by ORF3 coded by fap1-orf4 gene locus of Streptococcus parasanguis
Yirong LI ; Xiang HUANG ; Hui WU ; Lihua HU
Chinese Journal of Microbiology and Immunology 2009;29(5):460-465
Objective To study whether ORF3 coded by fap1-orf4 gene locus of Streptococcus pa-rasanguis is involved in the regulation of Fap1 glycosylation and maturation and to investigate whether ORF3 influences Streptococcus parasanguis adhesion. Methods A gene replacement strategy was adapted to con-struct orf3 alleic replace mutant of Streptococcus parasanguis. Complementation assay and Western blot were used to test Fap1 expression levels. Whole saliva-coated hydroxyapatite (SHA) adhesion assay was adapted to determine Streptococcus parasanguis adhension. Results (1) Non-polar was found in strain VT1774, the orf3 alleic replace mutant of Streptococcus parasanguis. (2) Western blot showed that mature Fapl (Mr about 220 × 103) disappeared and were substituted with high molecular weight Fapl (Mr about 470 × 103) in strain VT1774, furthermore, complementation assay showed VT1775, the complementation strain of VT1774, re-stored mature Fapl expression. (3) The binding ability reduced significantly in strain VT1774. Conclusion ORF3 coded byfapl-orf4 gone locus was required for Fap1 glycosylation and maturation in Streptococcus pa-rasanguis, orf3 alleic replacment resulted in Fap1 glycosylation and mature disorder and decreasing of adhen-sion ability of Streptococcus parasanguis.
8.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.
9.Optimizing extraction process of Chonglian oral solution by orthogonal test
Yongzhong WANG ; Lunzhu XIA ; Yirong FENG ; Li CHEN ; Mei MENG
Chinese Journal of Primary Medicine and Pharmacy 2011;18(4):436-438
ObjectiveTo optimize Chonglian oral solution extracting craft. MethodsWith the obtaining rate of extract and the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the water extraction process,and with the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the alcohol deposition process,orthogonal design was used to optimize the conditions for the extraction process respectively. ResultsThe optimal conditions for the water extraction of Chonglian oral solution was as following:to add water 10 times,decocting 3 times,1 hour each time.The optimal conditions for the alcohol deposition of Chonglian oral solution was as following:concentrated for the relative density to 1.10(80C hot test),cold,add ethanol to the solution for the ethanol content of the solution reached 80%,static settlement for 24 hours. ConclusionThe extracting method is reasonable,stable,and suitable to industrialized producting.
10.Effects of BMI-1 over-expression on HOX family expression and cell cycle in HeLa cells
Fenghua CHEN ; Yirong LI ; Lin WANG ; Lihua HU
Chinese Journal of Pathophysiology 2009;25(12):2366-2370
AIM: To observe whether transfection of mammalian expression vector pEGFP containing the gene of B-cell specific moloney leukemia virus insertion site 1(BMI-1) could express in human cervix cancer cell line HeLa, and to detect its effect on HOX family expression and cell cycle.METHODS: pEGFP-BMI-1 was transfected into HeLa cells with Lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blotting. SYBR green I real-time RT-PCR was used to quantitate mRNA expression of P16~(INK4a), hTERT, HOXA9, HOXB4 and HOXC13. FACS analysis was used to detect the change of cell cycle.RESULTS: In HeLa cells transfected with pEGFP-BMI-1, the results of real-time RT-PCR showed that the mRNA expressions of P16~(INK4a), HOXA9 and HOXC13 were reduced to 9.2%, 10.9% and 69.7%, respectively, as compared to control HeLa cells (P<0.01). However, hTERT and HOXB4 mRNA expressions did not change significantly (P>0.05). FACS analysis showed a decrease from 65.68 % to 50.53% in G_1 population and a significant increase from 27.17% to 39.59 % in S population after transfection (P<0.01).CONCLUSION: BMI-1 over-expression in HeLa cells down-regulates mRNA expressions of P16~(INK4a), HOXA9 and HOXC13, decreases G_1 population and increases S population. Therefore, BMI-1 may be involved in carcinogenesis and cancer development.