Comparison of molecular diagnosis with serum markers and synovial fluid analysis in patients with prosthetic joint infection.

AIMS: The aim of this study was to compare the results of 16S/28S rRNA sequencing with the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, and synovial fluid analysis in the diagnosis of prosthetic joint infection (PJI). PATIENTS AND METHODS: Between September 2015 and August 2016, 214 consecutive patients were enrolled. In the study population, there were 25 patients with a PJI and 189 controls. Of the PJI patients, 14 (56%) were women, and the mean age at the time of diagnosis was 65 years (38 to 83). The ESR and CRP levels were measured, and synovial fluid specimens were collected prospectively. Synovial fluid was subjected to reverse transcription polymerase chain reaction (RT-PCR)/sequence analysis targeting the 16S/28S rRNA, and to conventional culture. Laboratory personnel who were blind to the clinical information performed all tests. The diagnosis of PJI was based on the criteria of the Musculoskeletal Infection Society. RESULTS: A total of 25 patients had a confirmed PJI. In 20 cases of monomicrobial PJI, the PCR products could be perfectly matched with the 16S/28S rRNA genes specific for different species of bacteria provided by sequence analysis. Of the five polymicrobial cases of PJI, 16S/28S rRNA PCR sequence analysis failed to identify the concordant bacteria species. In the 189 control patients, there was one false-positive RT-PCR result. The sensitivity and specificity of the molecular diagnosis method were 100% (95% confidence interval (CI) 85.7 to 100) and 99.5% (95% CI 97.1 to 99.9), respectively, whereas the positive and negative predictive values of PCR were 96.1% (95% CI 79.6 to 99.9) and 100% (95% CI 98.1 to 100), respectively. The PCR results were significantly better than serological diagnostic methods (p = 0.004 and p = 0.010 for ESR and CRP, respectively), the synovial fluid white blood cell (WBC) count (p = 0.036), and percentage of polymorphonuclear cells (PMN%) (p = 0.014). CONCLUSION: Stepwise RT-PCR and sequence analysis of the 16S/28S rRNA carried out under stringent laboratory conditions achieved highly sensitive and specific results for the differentiation between aseptic and septic joints undergoing arthroplasty. Sequence analysis successfully identified bacterial strains in monomicrobial infections but failed to identify molecular targets in polymicrobial infections. Further refinement of the protocols to identify the bacteria in polymicrobial infections is needed. Cite this article: Bone Joint J 2018;100-B:1345-51.

Integrated bioinformatics analyses identify dysregulated miRNAs in lung cancer.

BACKGROUND: Lung cancer is the most common cause of cancer-related death worldwide. MicroRNAs (miRNAs) play important roles in various biological processes, including cell development, proliferation, differentiation and apoptosis. MATERIALS AND METHODS: In the current study, using miRNA expression profiles from the Gene Expression Omnibus (GEO) database, we used three independent tests: Wilcox test, t-test and Fisher's exact test to investigate miRNA's involvement in lung carcinogenesis. RESULTS: Ten differentially expressed miRNAs were identified. Among them, miR-675 drew specific attention. Ingenuity pathway analysis of its target genes revealed its impact on cell death and cell cycle. It is possible that miR-675 contributes to the pathogenesis of lung cancer through its down regulation of the tumor suppressor gene RB1. CONCLUSIONS: Our results suggest miR-675 may serve as a potential therapeutic target of lung cancer.

Generation of induced pluripotent mouse stem cells in an indirect co-culture system.

Typically, production of induced pluripotent stem cells requires direct contact with feeder cells. However, once the stem cells have reached the appropriate maturation point, it is difficult to separate them from feeder cells, which must be irradiated with γ-rays or treated with the antibiotic mitomycin-C. We used a microporous poly-membrane-based indirect contact co-culture system with mouse embryonic fibroblasts to induce mouse pluripotent stem cells without radiation or antibiotics. We found that induced pluripotent stem cells induced by this co-culture method had a reprogramming efficiency and time similar to those induced using traditional methods. Furthermore, strongly expressed pluripotent markers showed a normal karyotype and formation and contained all three germ layers in a teratoma.

Frequent allelic loss at the FRA3B site in endemic nasopharyngeal carcinoma: association with clinical features and Epstein-Barr virus infection.

In this study, we aimed to precisely define the patterns of allelic loss at the FRA3B site in endemic nasopharyngeal carcinoma and to determine whether an association exists between allelic loss, clinicopathological features and Epstein-Barr virus infection. We examined the loss of heterozygosity in 40 cases of nasopharyngeal carcinoma from an endemic area in southern China, using eight high dense, polymorphic, microsatellite markers within or flanking the FRA3B site. Loss of heterozygosity at the FRA3B region was shown in 31 (77.5 per cent) primary tumours. Loss of heterozygosity was found most frequently at the D3S1300 (55.6 per cent) and D3S2757 (50.0 per cent) loci. The common area of deletion was located between the D3S4103 and D3S4260 loci. In nasopharyngeal carcinoma, loss of heterozygosity at the FRA3B/fragile histidine triad locus correlated with the following clinicopathological parameters: tumour T-stage, lymph node status, clinical stage, tumour differentiation and serum antibody titres of immunoglobulin (Ig) A against Epstein-Barr virus capsid antigen. Significantly frequent loss of heterozygosity was observed in nasopharyngeal carcinoma with tumour stages T3 and T4, lymph node metastasis and advanced tumour-node-metastasis staging (III and IV). Very frequent loss of heterozygosity was also observed to correlate with World Health Organization type III nasopharyngeal carcinoma histopathology. We also found that nasopharyngeal carcinoma patients with high titres of IgA against Epstein-Barr virus capsid antigen showed very frequent loss of heterozygosity. Allelic loss at the FRA3B site occurs significantly more commonly in endemic nasopharyngeal carcinoma patients. This suggests that the region between D3S4103 and D3S4260 may represent a preferential molecular target in nasopharyngeal carcinogenesis.

Mutation and abnormal expression of the fragile histidine triad gene in nasopharyngeal carcinoma.

OBJECTIVE: To determine alterations of fragile histidine triad (FHIT) gene in nasopharyngeal carcinoma and the correlation of FHIT gene with nasopharyngeal carcinogenesis. STUDY DESIGN: Prospective study. METHODS: A total of 28 nasopharyngeal carcinoma and 16 normal nasopharyngeal epithelium specimens were examined for abnormalities of FHIT gene by nested reverse-transcriptase-polymerase chain reaction and DNA sequencing. RESULTS: The deletion of FHIT gene was not observed in 16 normal nasopharyngeal epithelium specimens. In 28 cases of nasopharyngeal carcinoma tissues, 12 (42.9%) exhibited FHIT aberrant transcripts. Complementary DNA sequencing revealed exonic deletion, small DNA insertion, synonymous mutation in exon 8, or frameshift mutation in exon 5. CONCLUSIONS: The present results suggest that the FHIT gene may play an important role in the pathogenesis of nasopharyngeal carcinoma and may be one of the candidate tumor suppressor genes in nasopharyngeal carcinoma.

[Treatment of epistaxis by radiofrequency].

OBJECTIVE: To evaluate the therapeutic effect of radiofrequency on epistaxis. METHOD: Three hundred and ten patients with epistaxis were treated with radiofrequency and Nd:YAG laser, respectively. RESULT: The effective rate of radiofrequency was 88.8%, Nd:YAG laser 90.5%. CONCLUSION: The therapeutic effects of epistaxis by radiofrequency and ND:YAG laser are similar, but radiofrequency is simpleier, easy to perform, more and useful than Nd:YAG laser.

Quantitative chromatics analysis for computer imaging of cytologic subtypes of lung cancer stained by Papanicolaou stain.

OBJECTIVE: To determine the role of quantitative chromatics analysis in the classification of subtypes of lung cancer stained by Papanicolaou stain. STUDY DESIGN: By means of computer image analysis, 60 keratinized squamous carcinoma cells (KSCC), 88 nonkeratinized squamous carcinoma cells (NKSCC) and 150 adenocarcinoma cells (ACC) from lung cancer in sputum smears stained by Papanicolaou stain were analyzed and distinguished based on quantitative colorimetry. The features measured were the content of three primary colors, red (R), green (G) and blue (B) and the coefficients of R, G and B (r, g and b, respectively). Hue, saturation, brightness and gray level were also measured. A stepwise discriminant analysis was carried out. RESULTS: The values of R, G and B and r, g and b, hue and saturation in NKSCC and ACC were significantly different from those of KSCC, and the changes in the three primary colors were more sensitive than those in the gray level. Computer assessment based on three primary color coefficients, hue and saturation yielded accuracy of distinguishing KSCC from NKSCC and KSCC from ACC of 95.2% and 95%, respectively. CONCLUSION: Quantitative analyses of R, G and B and r, g, b and hue and saturation are valuable in distinguishing KSCC from NKSCC and ACC.

[Benign paroxysmal positional vertigo following radiotherapy for nasopharyngeal carcinoma (report of 3 cases)].

OBJECTIVE: To investigate the effect of ionizing radiation on the vestibule of the inner ear. METHOD: Three patients with benign paroxysmal positional vertigo (BPPV) accompanied by hearing loss following radiotherapy for nasopharyngeal carcinoma (NPC) were reported. Particle repositioning maneuver (PRM) was used as treatment for BPPV, and salvia miltiorrhiza tablets were taken orally to treat hearing loss. RESULT: Two of three cases with BPPV recovered and one was effective; Mean phone frequency hearing increased 10 dB compared with that before therapy. CONCLUSION: Occurrence of BPPV in this series may be related to radiation damage on the vestibule; PRM is a effective modality for the patients with BPPV.

[Evaluation of the effect of streptomycin perfusion of the labyrinth in the treatment of Meniere's disease and endolymphatic hydrops].

OBJECTIVE: To investigate the clinical effect of streptomycin perfusion of the labyrinth (SPL) in the treatment of Meniere's disease. METHOD: 13 patients with Meniere's disease and 3 patients with delayed endolymphatic hydrops (DEH) underwent SPL. The follow-up time was between 5 and 7 years (average 6.3 years). RESULT: Vertigo was completely controlled in 9 patients, substantialy controlled in 4, limitedly controlled in 2 and not controlled in 1. Hearing was improved in 1 patients, stabilized in 5 and worse in 10. Tinnitus was improved in 3 patients, stabilized in 5 and worse in 8. CONCLUSION: SPL is a safe and effective method in the treatment of Meniere's disease and DEH, but the hearing may be insulted and tinnitus may be worse in some patients.

[Low volume mechanical ventilation in the treatment of patients with chronic obstructive lung diseases with type II respiratory failure].

To study the value of low volume ventilation in chronic obstructive lung disease (COPD) with Type II respiratory failure, 35 patients were divided into two groups at random. The tidal volume of regular volume ventilation group was 12-14 ml.kg-1 and that of low volume ventilation group was 6-8 ml.kg-1. Some indexes of airway pressures and blood gases were recorded and those of cardiovascular function were monitored continuously. The results were that compared to the normal volume ventilation group, the actual ventilation in liters per minute was lower obviously, the expiratory time longer, the incidence rate of lung barotrauma lower, peak inspiratory pressure, end-inspiratory-pressure, and internal positive-end-expiratory pressure smaller, and side-effect on hemodynamics less in the low volume group. The results suggest that low volume ventilation with permissive hypercapnia is better than normal volume ventilation in COPD.

[Detection of D -dimer in pleural effusion].

The content of D -dimer in pleural fluid of different causes was examined by the ELISA method in 45 patients. The results were that the contents of D -dimer in both tuberculous and empyema pleural effusions were significantly higher than in malignant pleural effusion (P < 0.01; P < 0.05). D -dimer was positively correlated with lactic dehydrogenase (LDH) in pleural fluid (r = 0.4168, P < 0.01). The results suggest that fibrinolytic activities in tuberculous and empyema pleural effusion are much higher than in malignant pleural effusion. The detection of D -dimer in pleural fluid may reflect the extent of pleural inflammation and be helpful in the differential diagnosis of pleural effusion.

[The relation between p53 expression and lymph node metastasis of breast cancer].

The monoclonal antibody to p53 and a rapid immunohistochemical method were used in this study to detect p53 expression in 50 paraffin-embedded breast cancers and 7 benign hyperplastic diseases of breast. The results of immunostaining showed that 12 (24%) cases of breast cancers were positive and all benign lesions were negative. No significant difference was found between primary and metastatic sites of 11 paired samples of breast cancers in p53 expression, suggesting that the abnormal expression of had p53 occurred before lymph node metastasis of breast cancer. Statistical analysis revealed that the abnormal expression of p53 was significantly correlated with lymph node metastasis of breast cancer (P < 0.005) and also correlated with medullary breast cancer in which two of four cases strongly positive to p53 had lymph node metastasis. Our results indicate that the abnormal expression of p53 is not only involved in the development, but also involved in the metastasis of breast cancer. Therefore, detection of p53 expression may have significant clinical importance in the evaluation of the tendency of lymphatic metastasis of breast cancer.

Tube-perilymph fistula. An experimental study.

Before exploratory tympanotomy, it is difficult to diagnose an idiopathic perilymph fistula (PLF) because of its variable presentations. An animal model tube-PLF on guinea pigs was established to evaluate the change of the sensitivity of the cochlea function and the value of the postural audiometry in PLF. A fine polyethylene tube (O.D. 0.6mm) was inserted into the scala tympani of the basal turn through the incised round window membrane in 10 experimental guinea pigs, and another 5 as controls. The hearing threshold (HT), action potential (AP) latency for stimulus of 105 dB SPL, difference of HT and AP latency from the test ear down to up were measured by electrocochleography (ECoG) immediately after operation and one week later. The HT, AP latency and postural audiometry showed no significant changes immediately but a significant difference one week later in HT (P less than 0.01) and postural audiometry, the AP latency also presented a significant delay (P less than 0.01). The results showed that the tube-PLF model may be a valuable method for studying chronic PLF.