Francisella guangzhouensis was used as an example to illustrate how to utilize the bioinformatics resources to identify the novel species, how to name the novel species with Latinized letters, and how to validate a novel species in the clinical microbiology laboratories.Some related rules of International Code of Nomenclature of Bacteria was also introduced.

OBJECTIVE To develop a new rapid method of real time fluorescence quantitative Polymerase chain reaction(PCR) to detect Polyoma virus BK(BKV) with high sensitivity,specificity,stability and lower price using the self-quenched probe.METHODS DNA segment of BKV was cloned into the vector pGEM-11Zf.The recombinant vector pGEM-11Zf-BK was used as standard template.The self quenched probe was designed according to the cloned gene sequence.The PCR reaction system was optimized and the method established was evaluated.RESULTS Restriction maps showed that the structure of recombinant plasmid pGEM-11Zf-BK was the same as expected.Methodology analysis suggested that self-quenched probe fluorescent quantitative PCR detective method of BKV be successfully established with high sensitivity,specificity and accuracy.CONCLUSIONS The self-quenched probe fluorescence quantitative PCR detective method for BKV has been developed successfully with high sensitivity,specificity and accuracy.

Objective To discuss the comparability of TSH results among different detection systems.Methods 4 different kinds of chemiluminescence immune detection systems (Bayer ACS180, Bayer CENTAUR, VITROS ECI and DPC immulite-1000) were used to detect TSH concentration in 3 kinds of quality control specimens (Bayer,DPC and self-made) and clinical sera. The collected data were dealt with statistical analysis.Results Analysis of variance showed the TSH results from different control and patients sera had significant difference in different detection systems (P

Objective To identify one acid‐fasting bacteria isolated from wound secretion of breast abscess .Methods The acid‐fasting strain was identified by the morphological characteristics ,API 20A strips ,classical biochemical reaction ,and 16S rRNA gene sequencing .Results Cells of the strain was anaerobic ,non‐spore‐forming ,pleomorphic ,straight or curved rods ,which Gram and acid‐fast stain both were positive .After incubation for 5 days on sheep blood agar ,colonies were than 2 mm in diameter ,circular , smooth ,entire ,bump ,rice cream‐like withβ‐hemolysis .The 16S rRNA gene sequences were 100% identity to Propionibacterium av‐idum .The API 20A profile was 44365062 with positive Voges‐Proskauer test ,which was also consistent to Propionibacterium avi‐dum .Conclusion The pathogens of breast abscess is Proionibacterium avidum ,which is the first acid‐fasting Propionibacterium re‐ported in China .

OBJECTIVE To investigate ?-lactamases genes in multi-resistant Pseudomonas aeruginosa in Guangzhou. METHODS We collected 22 P.aeruginosa strains from hospitalized patients,and detected eleven types of ?-lactamases associated genes by PCR. RESULTS CARB,IMP,TEM,VEB and VIM showed positive amplifications with positive proportions were 100.0%,95.5%,77.3%,13.6% and 4.5%,respectively.Other genes such as GIM,SPM,GES,PER,DHA,OXA-10 and SHV showed negative results. CONCLUSIONS The genes CARB and IMP have high positive rate.Different types of ?-lactamases associated genes in P.aeruginosa aften lead to multi-resistance to antibiotics,which bring great difficulties to clinical therapy and hospital infection monitoring.

ObjectiveTo investigate the effect of basic fibroblast growth factor-polyactide release nanospheres on proliferation and adipogenic induction of adipose-derived stem cells in vitro.MethodsAdipose-derived stem cells were isolated and induced for three-line differentiation in vitro.The culture medium and inductive medium of stem cells were prepared containing 0,1,2,3,4 and 5 mg/ml basic fibroblast growth factor-polyactide release nanospheres,respectively.Adipose-derived stem cells were cultured ina 96-well plate and replaced the culture medium containing release nanospheres the second day.The cells proliferation was detected by the method of MTT every day and quanti fication of oil red O every other day.The data obtained were analyzed with SPSS13.0 statistical software.ResultsThe basic fibroblast growth factor polyactide release nanospheres had the ability promoting proliferation and adipogenic induction of adipose-derived stem cells.The best concentration of nanospheres was 3 mg/ml and 4 mg/ml,respectively.ConclusionsThe basic fibroblast growth factor-polyactide release nanospheres could promote proliferation and adipogenic induetionof adipose-derived stem cells significantly.It could be used as an ideal cytokine release nanospheres in adipose tissue engineering.

Objective To investigate the survival profile of the intradermally injected mouse autologous skin fibroblasts and the changes of the collagen fibers by using green fluorescent protein labeling and two-photon fluorescence microscopy. Methods The cultured cells were transfected by EGFP lentivirus, and then the cells were injected into the corresponding mouse skin. Biopsy was taken from the animals after 1 and 2 months. The specimens made serial frozen sections, the survival profile of the injected cells and the changes of the collagen fibers were observed by two-photon fluorescence microscopy. The collagenic area and dermal thickness were measured with image analysis software, and statistical analysis was also carried out. Results Two-photon fluorescence microscopy showed clear images of the injected cells and collagen fibers. Both the area of collagen fibers and the dermal thickness were significantly increased in injected cells after 2 months (P<0.05), however, there were no difference between injected cells and control after 1 mouth (P>0.05). Conclusions Autologous cultured fibroblasts could survive in a long time after transplantating into the skin, and collagen could be newly produced, the depth of dermis increases, which provides a possibility to treat mini-defects of the tissue.

Objective To indentify Streptobacillus moniliformis isolated from the knee joint pus by 16S rRNA gene sequencing and biochemical reactions and explore the clinical value of the method. Methods The bacterial 16S rRNA gene sequence-based identification, bacterial morphology, VITEK 2 automate systems, API 20NE strips, API 20E strips and API 50CH were performed to identify the rare bacteria. Results The bacteria grew slow on blood agar and chocolate agar and were inhibited on Maconkey agar. The bacterial colony on blood agar tookes the form of 1~2 mmomelette, which was translucent and moist with circular protrusion and smooth edges. They were Gram-staining negative and in catenation, its thalli 1~3μm, round, oval or fusiform. Vitek 2 GN-13, API 20NE and API 20E were unable to reach the identification of the bacteria. 16S rRNA gene sequencing showed the bacteria were similar to streptobacillus moniliformis by 100%. Conclusion The rare bacteria isolated from left knee joint are streptobacillus moniliformis. 16S rRNA gene sequences combined with the biochemical reactions is accurate in the identification of these bacteria.

Objective Parainfluenza virus is an important pathogen of lower respiratory tract infections in infants and young children.This study was to search for a method for rapid culture and identification of human parainfluenza viruses from nasal swabs. Methods Nasal swab specimens were collected from 0-5 years old children with acute respiratory tract infection.The specimens were inoculated onto 96 plates with prefabricated LLC-MK2 cells and then centrifuged for 1 hour at 3000 r/min and also inoculated using the traditional culture method, followed by addition of virus mainte-nance medium containing 4 μg/mL TPCK trypsin.The cytopathic effect was observed daily, and hemagglutination and blood absorption tests were done at 2, 5, and 8 days after inoculation.In case of posi-tive result of either test, the specimen was subjected to immunofluo-rescence staining. Results Six strains of parainfluenza virus were isolated from the 83 nasal swab specimens, with a positive rate of 7.2%.There was a significant difference in the rate of separation be-tween the rapid and traditional culture methods after 2 days of culturing (7.2%vs 0%, P<0.05).The infected cells produced a cy-topathic effect that characterized by syncytium and crush formation.Hemagglutination and blood adsorption tests were positive at 4℃and negative at the room temperature.Immunofluorescence staining exhibited specific apple green fluorescence. Conclusion The method for rapid culture and identification of human parainfluenza viruses in nasal swab specimens was successfully established, which can be used to obtain and identify parainfluenza viruses with virulence and biological activity in 2 days.

Objectives To identify and characterize 10 strains of Francisella philomiragia-like organisms isolated from blood samples and environmental water.Methods The 10 clinical and environmental isolates were identified by traditional morphological examination and biochemical characterization,matrix-assisted laser desorption/ionization time of flight(MALDI-TOF) mass spectrometry(MS) systems and sequencing based on 16S rRNA gene.The minimum inhibitory concentrations were tested by E-test methods.Results All the 10 isolates were gram-negative coccobacilli appearing tiny and faint counterstain of safranin,negative for urease,nitrate reduction and X and/or V factor requirement,but positive for oxidase and catalase.The isolates grew rapidly in sheep blood agar,chocolate agar and BCYE plate forming white opaque,colorless transparent or gray smooth colonies with about 2-mm diameters,but did not grow in M-H agar and MacConkey agar.The sequencing for 16S rRNA gene indicated that the 10 isolates shared more than 99.6％ similarity to Francisella philomiragia,and fell into the same clusters of Francisella philomiragia on phylogenetic tree.The MALDI-TOF MS analysis also showed the typical peaks with 6 153 m/z,5 180 m/z,7 757 m/z and 9 392 m/z which were similar to Francisella philomiragia ATCC 25015.However,they may be misidentified to be Sphingomonas paucimobilis by using Vitek 2 GN cards,Neisseria cinerea by using Vitek 2 NH cards,Myroides odoratimimus by using API 20NE strips and Haemophilus by using API NH cards.The results of antimicrobial susceptibility showed that they were all sensitive to chloramphenicol,doxycycline,tetracycline,gentamicin,ofloxacin and ciprofloxacin.Conclusion The 10 isolates could be identified as Francisella philomiragia,so we should pay more attention to the infrequent pathogen for its inactive biochemical reaction and the misidentification by commercial detection systems.