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.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.
3.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.
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.Study on the TIM-4 gene polymorphism and its association with the susceptibility to allergic asthma in Chinese Ban population of Hubei province
Xiaohui WU ; Yirong LI ; Qingguo CHEN ; Fenghua CHEN ; Lihua HU
Chinese Journal of Microbiology and Immunology 2008;28(9):833-837
Objective To investigate whether two polymorphism sites of the the exon 2 Lys65Lys(G/A) and exon 9 Va1365Met(G/A) in T ceils immunoglobulin domain and mucin domain protein-4(TIM-4) are associated with asthma in Chinese Han population of Hubei province. Methods The polymorphisms were de-tected with polymerase ehain reaction-restriction fragment length polymorphism(PCR-RFLP) in 185 cases of al-lergic asthma and 162 healthy controls. The genotype and allele frequencies were calculated and analyzed. Re-sults(1)The genotype frequencies of G/G, G/A and A/A in Lys65Lys(G/A)polymorphism were 0.840, 0.160 and 0 respectively in the healthy population, and were 0.859, 0.141 and 0 respectively in the allergic asthma population. No significant difference in genotype and allele frequencies was found between asthma pa-tients and the control subjects (P=0.603, P=0.618). (2) The polymorphism of the Val365 Met(G/A) was not detected in our study. Conclusion There is polymorphism site of the exon 2 Lys65Lys(G/A)in TIM-4, but this polymorphism site is not associated with asthma in Han nationality in Hubei Chinese population. There is no SNP of the exon 9 Val365Met(G/A) in TIM-4 in Chinese Han population of Hubei province.
6.Study on function and interaction of GTF and ORF4 coded by fap1-orf4 gene locus of Streptococcus parasanguis
Yirong LI ; Lihua HU ; Xiang HUANG ; Hui WU
Chinese Journal of Microbiology and Immunology 2008;28(9):771-776
Objective To investigate whether glucosyltransfernse(GTF) and open reading frame 4 (ORF4) coded by fap1-orf4 gene locus of Streptococcus parasanguis was involved in the regulation of Fap1 glycosylation, and mature and to determine whether there was interaction between GTF and ORF4. Methods A gene replacement strategy was adapted to construct gtf and orf4 allele replace mutant of S. Parasanguis. Complementation assay and Western blot were used to test fap1 expression levels. Yeast two-hybrid analysis and GST pull down assays were adapted to determine the interaction between GTF and ORF4. Results (1) Compared with wild S. Parasanguis, mature Fapl (Mr about 220×103) disappeared and were substituted with high molecular weight Fapl (Mr about 360×103) in gtf or orf4 alleie replace mutants of S. Parasan-guis. Complementation assay showed that pVPT-GFP-gtf and pVPT-GFP-orf4 restored mature fap1 expression in gtf or orf4 alleie replace mutants, respectively. (2) With Yeast two-hybrid analysis, the eotransformants, AH109/pAD-Gtf+pBD-orf4 and AHlOg/pAD-orf4+pBD-gtf growed on SD-LTHA selective ngar plate after streaked, reversely, the eotransformants, AH109/pAD+pBD-orf4,AH109/pAD+pBD-gtf、AH109/pBD+ pAD-orf4、AH109/pBD+pAD-gtf did not grow on SD-LTHA selective agar plate, furthermore, the cotrans-formants, AH109/pAD+pBD-orf4 and AH109/pAD-orf4+pBD-gtfshowed blue during X-α-gal assay. (3) GST pull down assay confirmed the direct interaction between GTF and ORF4. Conclusion There is inter-action between GTF and ORF4 coded byfapl-orf4 gene locus of S. Parasangnis and the formation of the GTF and ORF4 complex was required for the glycosylation and mature of Fapl in S. Parasanguis.
7.Detection of telomerase gene expression In the peripheral blood mononuclear cells in patients with colorectal carcinoma by real-time quantitative telomeric-repeat amplification protocol assay
Changxin SHEN ; Lihua HU ; Lin XIA ; Yirong LI ; Guohong LIN
Chinese Journal of Laboratory Medicine 2008;31(8):880-883
Objective To investigates the gene expression of telomerase by real-time quantitative telomeric-repeat amplification protocol assay (RTQ-TRAP) in the peripheral blood mononuclear cells (PBMC) of colorectal carcinoma patients and the relationship between the telomerase gene expression in PBMC and clinicopathological features. Methods Peripheral blood samptes were collected from 71 colorectal carcinoma patients, 20 benign colorectal disease patients and 25 normal controls. The telomerase gene expression in PBMC was measured by RTQ-TRAP, and serum CEA in colorectal carcinoma patients was measured by chemiluminescence immunoassay. Results The gene expression of telomerase in PBMC was positive in 50 out of 71 cancer patients (70. 4% ) , 1 out of benign patients (5. 0% ), respectively. The difference of the telomerase gene expression of PBMC in benign colorectal diseases and cancer patients was significant (χ2 = 24. 521, P < 0. 001 ). There was no significant association between the expression of telomerase and patient's gender, age, Dukes stage, and tumor site. The positive rate of CEA in colorectal carcinoma patients was not significantly higher than positive rate of telomerase gene expression(χ2 = 2. 286,P = 0. 125). Conclusions The RTQ-TRAP method is highly accurate and sensitive in measuring telomerase gene expression. The detection of telomerase gene expression in PBMC of colorectal carcinoma patients is a simple and useful molecular marker for the diagnosis of colorectal carcinoma.
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.Evaluation of a modified Chinese version of the National Institutes of Health-Chronic Prostatitis Symptom Index
Yifu LI ; Hui XIE ; Yu YANG ; Yong CAI ; Yirong YANG
Chinese Journal of Urology 2010;31(10):710-714
Objective To revise the Chinese version of the National Institutes of Health-Chronic Prostatitis Symptom Index (CHN-NIH-CPSD), and evaluate its feasibility, reliability, validity and responsiveness. Methods The NIH-CPSI was translated into Chinese according to a standard methodology including forward-backward-forward technique. The CHN-NIH-CPSI was pre-tested in consecutive samples of 162 native-speaking Chinese chronic prostatitis(CP)patients. Ninety-five of 162 filled the index again on the same day and after 4-week therapy. Ninety-seven healthy men were included as evaluated. Results The recovery of the questionnaires was 100% and all the patients filled the index completely. The mean time to complete the questionnaire for the patient group was 5.2±2.4 (range 2 - 12) min. The split-half reliability was 0.82. For the overall index and each subscale, the test-retest reliability was 0.98, 0. 98, 0. 98, 0. 97, respectively(P<0.01);and the Cronbach's α coefficient was 0. 61,0. 71, 0. 59, 0. 75, respectively. The confirmatory factor analysis showed good construct validity with a goodness of fit index of 0. 85 and a x2 of 124.67(P<0. 01). Of all 162 patients, the scores of the overall index and each subscale were 23. 33±5.91. 8. 80±4.26, 5.30±2.82, 9. 23±1.90, respectively;and those of healthy controls were 1. 95±1.97, 0. 37±1.03, 0. 15±0.58, 1.42± 1.20,respectively. Of the 95 patients, the original scores were 23. 53±5.60, 9.21 ±4.04, 5.10±2.75,9.21 ±2.05, comparing with 19.47±6.36, 7.79±3.95, 3. 58±1.88, 8.11±2.50, the 4 weeks later scores. The group t-test and paired t-test showed good responsiveness. Conclusions The CHN-NIH-CPSI has high feasibility, reliability, validity and responsiveness for testing the patients with CP. It is suitable for Chinese-speaking patients and helpful for cross-cultural comparisons of men with CP in clinical and research settings.
10.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.