[Experimental studies on chitosan blend membrane as scaffold carriers for cultivating rabbit corneal endothelial cells in vitro].

Un-transfected rabbit corneal endothelial cells (RCECs) were cultivated, using chitosan blend membrane 4ha (chitosan-hyaluronic acid), 631ha (chitosan-hyaluronic acid) and 631s (chitosan-chondroitine sulfate) as scaffold carriers. Their biocompatibilities were studied in regard to cell adherence, morphological changes, growth status and monolayer forming abilities. The results indicated that RCECs cultivated on 4ha and 631ha carriers tended to be aggregated and even desquamated to some extent in local areas, and even more severely on 631ha carrier. And the RCECs cultivated on 631ha carrier could form almost a monolayer 48h later, and those on 4ha carrier could not. Contrarily, the RCECs cultivated on 631s carrier were evenly distributed and were in good status of growth with a good adherence and fibroblast-like morphology which could form almost a monolayer 48h later. And a complete monolayer was formed and was tightly attached to the 631s carrier 72h later. From the above results, it can be concluded that 631s carrier is most probably an ideal scaffold carrier for RCEC cultivation. 631s carrier may have the potential for use in the development of tissue-engineered rabbit corneal endothelium.

Detection and identification of viral pathogens in patients with hand, foot, and mouth disease by multilocus PCR, reverse-transcription PCR and electrospray ionization mass spectrometry.

BACKGROUND: Rapid detection and identification of viruses are important for early diagnosis and effective surveillance of hand, foot, and mouth disease (HFMD). We described a novel assay using multilocus PCR and reverse transcription-PCR coupled with electrospray ionization mass spectrometry (RT-PCR/ESI-MS) to simultaneously detect and identify human enterovirus A-D, adenovirus A-F, human herpesvirus 1-8, parvovirus B19 and polyomavirus. OBJECTIVES: To evaluate the accuracy and efficacy of the RT-PCR/ESI-MS method, to detect and type enterovirus from specimens of clinical diagnosed HFMD patients. STUDY DESIGN: In this study, 152 specimens of clinically diagnosed HFMD patients were studied by the assay using RT-PCR/ESI-MS method. The detection and typing of enterovirus by RT-PCR/ESI-MS were compared with results from reference molecular methods. RESULTS: The assay detected enteroviruses in 97 (63.8%) specimens, resulting in a sensitivity of 93.8% (95% CI: 91.8-95.7%) and a specificity of 87.5% (95% CI: 84.8-90.2%) compared with a reference clinical diagnostic test. Most enterovirus genotypes (65/84; 77%) determined by the platform were consistent with 5' UTR sequence analysis, and most misidentifications resulted from the virus library, which could be further improved by updating the enterovirus database. In addition to enteroviruses, herpesviruses, polyomaviruses, adenoviruses and human rhinoviruses were detected and identified in 55 (36%) HFMD specimens by RT-PCR/ESI-MS. CONCLUSION: With the capability of high throughput and detection and typing of multiple clinically relevant viruses simultaneously, RT-PCR/ESI-MS can be a rapid and robust laboratory tool for identifying viral pathogens.