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.

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.

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.

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.

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.

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.

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.

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.

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.

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.