Objective: The genetic characteristics of the human adenovirus type 53 (HAdV-53) strains isolated from Taiyuan city of Shanxi Province were studied to obtain the baseline data of their molecular characteristics. Methods: Conjunctival swabs (n=79) were collected from epidemic keratoconjunctivitis (EKC) patients in Shanxi eye Hospital in 2016, and five HAdV-53 strains were obtained after virus isolation and identification based on the three major capsid genes sequences including Penton base, Hexon and Fiber gene. And the corresponding sequences of global epidemic HAdV-53 strains and the strains with the same genetic origin as HAdV-53 were also downloaded from GenBank database, and then the three gene database were established, respectively. With the database, phylogenetic tree was constructed, and the genetic and molecular evolutionary characteristics were analyzed with bioinformatics software. Results: Five HAdV-53 strains in Shanxi Province in 2016 showed high consistency with the HAdV-53 strains prevalent in other countries in 1996-2014 (>99.8%). All HAdV-53 strains were in the same evolutionary branch with their recombinant source genotypes (HAdV-37 and HAdV-8) in Penton base and Fiber gene, respectively, and maintained a high degree of consistency in gene sequences. In Hexon gene, HAdV-53 strains were more closed to its recombinant source genotype HAdV-22, the nucleotide and amino acid sequences between two types were highly homologous, while HAdV-53 and HAdV-22 belonged to different evolutionary branches, and the evolution rate of HAdV-53 based on Hexon gene was 3.51×10^-5 substitution/site/year. Conclusion HAdV-53 has become an important new ocular infectious pathogen of Taiyuan. HAdV-53 strain are relatively conservative and stable based on Penton base, Hexon, and Fiber gene.

Objective To compare the difference of three methods testing the antibiotic susceptibility of mucoid Pseudomonas aeruginosa in order to provide accurate and reliable antibiotic susceptibility result for clinic.Methods A total of 630 mucoid Pseudomonas aeruginosa were collected from Linyi People′s Hospital during January 2015 to December 2016.They mainly come from respiratory medicine and the most common specimen source was sputum.All specimens were examined in 2 h.The strains isolated from the same patient were discarded.Antibiotic susceptibility was tested by the automatic microorganism analyzer VITEK2 compact, E-test, which was reference method, and K-B disk.The results of three methods were analyzed and compared by χ2 test.Results The result of E-test showed that antibiotic sensitivity of 630 mucoid Pseudomonas aeruginosa was above 52.7% except for Cefepime (39.2%).The result of K-B disk was compared with E-test, the antibiotic sensitivity of mucoid Pseudomonas aeruginosa to imipenem (72.4% vs 52.7%) and amikacin (48.6% vs 71.1%)had significant difference (χ2=8.283 7 and 10.533 8, P<0.05).The result of VITEK2 compact showed that the antibiotic susceptibility of mucoid Pseudomonas aeruginosa to imipenem(70.8% vs 52.7%), cefepime(60.8% vs 39.2%), gentamicin (87.6% vs 74.1%)and levofloxacin(81.3% vs 65.4%) was significant higher than the result of E-test (χ2=6.935 2,9.331 2,5.885 6 and 6.466 5, P<0.05).For tobramycin, piperacillin/tazobactam and ciprofloxacin, the result of three methods is more consistent.Compared to VITEK2 compact, the consistency between K-B disk and E-test was higher.The rate of very major error and major error were between 0.0%-4.8% (Amikacin 12.2%) and minor error was 4.6%-20.3%.Conclusions The drug sensitivity of mucoid Pseudomonas aeruginosa is different between various methods.The result of K-B disk and E-test using blood MH is more reliable than VITEK2 compact.

OBJECTIVE:To study pharmacognosy of Melilotus officinalis.METHODS:Qualitative identification of M.officinalis was conducted in respects of morphology,macroscopic characters,microscopic identification,UV spectra for medicinal extracts and IR spectra for medicinal powder.RESULTS:Nineteen bundles of rays were distributed in the radial bundle of root cross section of M.officinalis.The main vein vascular bundle of leaf cross section had a palisade tissue passing over it.The pith of the cross section of the stem occupied 4/5 of the whole cross section.The cross section of the leafstalk was heart-shaped,and unequal vascular bundles arranged in a triangular array.There were glandular hairs,which consist of three cells,and unicellular non glandular hairs in leaf epidermis.The crystal fibers of calcium oxalate crystals and infinitive blowhole were found in the leaves;glandular hairs and non glandular hairs could be found in the leafstalk under tissue dissociation;verrucous like protuberance and threaded catheter were found on the surface.The UV spectrum of extract and two order derivative IR spectrum of the medicinal powder showed obvious characteristics.CONCLUSIONS:Established method can be used for pharmacognostic identification of M.officinalis.

Objective To analyze the antibiotic resistance of Branhemella catarrhalis strains isolated from sputum specimens of patients with lower respiratory tract infections from Linyi, Shandong Province, and to explore the relationship between bro genotypes of the strains and their resistance to antibiotic agents.Methods Sputum specimens were colleted from the patients with lower respiratory tract infections in Linyi People ’ s Hospital from the January 2010 to December 2014.The specimens were inoculated into 4 different disks for bacterial isolation and cultivation.β-lactamase detection and drug sensitivity tests were performed, and PCR coupled with restriction endonuclease analysis was employed for bro genotyping.χ2 test was used to compare drug resistance of strains with different bro genotypes.Results A total of 497 Branhemella catarrhalis strains were isolated in five years, among which 221 strains were isolated in winter.All strains were sensitive to ertapenem and chloramphenicol, and the resistance rates to amoxicillin/clavulanate and cefaclor were low (≤2.8%).The strains were highly resistant to compound sulfamethoxazole, erythromycin and ampicillin (47.6%-89.8%), and there was a trend of increasing resistance rates with the year, but no statistically significant difference was observed ( P >0.05 ) .β-lactamases was positive in 412 strains (82.9%), and all of these strains were positive for bro gene, and the resistances to erythromycin, compound sulfamethoxazole, levofloxacin and ampicillin were higher in bro positive strains than those in bro negative strains (χ2 =12.16, 16.18, 8.41 and 200.00,P<0.05).Among bro positive strains, 391 (94.9%) were of genotype bro-1, 21 (5.1%) were of genotype bro-2, and their resistance to antibiotic agents was not of statistical difference ( P >0.05 ).Conclusions Most of Branhemella catarrhalis clinical isolates are β-lactamase producing strains, and bro-1 is the most common genotype.Strains are highly sensitive to carbapenems, cephalosporins andβ-Lactamaseinhibitors, which can be recommended for the treatment of Branhemella catarrhalis-related respiratory tract infections.

Objective To investigate the drug sensitivity of mucoid Pseudomonas aeruginosa to common antibiotics and the expression of β-lactamase-resistant phenotype.Methods The specimens were inoculated onto different disks to isolate and cultivate bacteria.The antibiotic susceptibility of mucoid Pseudomonas aeruginosa isolates was detected and judged by CLSI 2013.The detection of drug resistance was done by Kirby-Bauer (K-B) method and β lactamase-resistant phenotype was detected by E-test.SPSS19.0 was used to statistic data and x2 test was used to compare the antibiotic susceptibility between different groups.For all statistical test,a P values less than 0.05 was defined as statistically significant.Results The susceptibilities of mucoid Pseudomonas aeruginosa to the regular antibiotics were above 70％,of which the sensitivities to amikacin,to bramycin,gentamicin,imipenem and meropenem were higher than 90％.The positive rate of ampler class C β-lactamase (AmpC) was 28.3％ (56/198).The drug sensitivity of positive strains was lower than that of the negative strains,and the differentiation was significant to piperacillin-tazobactam,amikacin,ceftazidime,levofloxacin,ciprofloxacin and aztreonam (x2 =3.89-14.45,all P ＜0.05).The positive rate of extended spectrum β-lactamase(ESBLs) was 10.6％ (21/198).The drug sensitivity to ceftazidime and aztreonam of positive strains[42.9％ (9/21) and 57.1％ (12/21),respectively].It was lower than that of the negative strains [73.5％ (130/177) and 72.3％ (128/177)],x2 =5.06 and 19.24,both P ＜ 0.05.The difference of the other antibiotics was not significant(x2 =0.01-3.47,all P ＞0.05).The positive rate of metallo-β-lactamase (MBL) was 19.7％ (39/198),and the drug susceptibility of positive strains was lower than that of negative strains except gentamicin and aztreonam(x2 =4.07-15.99,all P ＜ 0.05).All the detected strains were Klebsiella pneumonia carbapenemase (KPC) negative.Conclusions The antibiotic susceptible rate of mucoid Pseudomonas aeruginosa was high,but some enzyme-produced strains were lower.The clinician should adjust medicine program by the results of laboratory.

Objective To discuss the distributional characteristics and drug-resistance of Enterococcus species isolated from urine specimens.Methods 3096 middle-segment urine specimens were collected since January to December in 2011 for culture.The identification of pathogenic bacteria and antibiotics sensitivity tests were carried out with VITEK2-compact combined with GN,AST-GN13,GP,AST-GP67,and antibiotics sensitivity tested performed by K-B and E-test at the same time.The results were determined by CLSI 2011.Results 1248 of 3096 pathogenic bacteria were isolated (40.31％).549 strains of Escherichia coli were detected (43.99％) which was the most common and 159 strains of Enterococcus were detected (12.74％) which was the second most common.The Enterococcus detection rate in woman (15.02％)was higher than that in man (10.35％),in out-patients (15.54％) than the that in hospitalized patients (12.49％),and in the patients of non-surgical departments (13.65％) than those of surgical departments (11.38％).The Enterococcus was absolutely sensitive to tigecycline,and the sensitive rate to vancomycin and linezolid were over 90％.The antibiotics sensitivity was higher for Enterococcus faecalis than that for Enterococcus faecium,and in surgical departments than non-surgical departments.Conclusions The detection rate of Enterococcus in urinary tract infection patients is quite high and varied between sexes and departments.The difference of drug resistance between species is obvious,and the bacteria species should be identified in order to use the antibiotics reasonably to postpone the development of drug resistant strains.

Objective To investigate the distribution and antibiotic susceptibilities of quantitatively cultivated bacteria from bronchoalveolar lavage fluid (BALF) samples. Methods Totally 312 BALF samples were streak inoculated to chocolate,blood and MAC plates with 10 μL annulus,and the bacterial colony ＞ 104 CFU/mL was considered pathogenic bacteria. The identification of pathogenic bacteria was carried out with Vitek 2-Compact,and Kirby-Bauer disc agar diffusion method,Etest and dilution method were used for antibiotics sensitivity test.Results Totally 216 (69.2％) strains of pathogenic bacteria were isolated.The major gram-negative strains were Pseudomonas aeruginosa,Acinetobacter baumannii,Klebsiella pneumoniae and Escherichia coli, and the major gram-positive strains were Staphylococcus aureus,Streptococcus pneumoniae and Staphylococcus epidermidis.The resistance rate of Pseudomonas aeruginosa to aztreonam was high,but lower than 30％ to piperacillin/tazobactam,imipenem,cefepime,ofloxacin,ceftazidime and amikacin.Staphylococcus aureus was highly resistant to erythromycin,benzylpenicillin and clindamycin,but it was sensitive to furadantin,vancomycin,quinupristin/dalfoprisdn,tigecycline and linezolid.Conclusion The positive rate of BALF cultivation is high,and the main pathogenic bacteria Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus and Escherichia coli are resistant to several commonly used antibiotics.

Objective To compare the drug resistance of mucoid Pseudomonas aeFagillosa with that of non-mucoid.Methods All specimens isolated and cultured from patients were identified and the antibiotic sensitivity was tested by automatic microorganism analyzer VITEK-32,GNI+,GNS-448,E-teat and K-B.Results Drug resistant rates of mucoid Pseudomonas aeruginosa to imipenem.levofloxacin.meropenem,cefepime,cefoperazone/sulbactam,amikacin,gentamicin,piperacillin.tazobactam,ceftazidime and cefotaxime were 5.3%, 16.1%, 5.6%, 20.6%, 1.1%, 10.5%,26.9%,5.3%.31.5% and 60.2%,respectively.The drug resistant rates of mucoid Pseudomonas aeruginosa were lower than those of non-mucoid except to ceftazidime.Conclusion Compared with non-mucoid Pseudomonas aeruginosa,antibiotic resistance of mucoid Pseudomonas aeruginosa is weaker in vitro.

BACKGROUND: Studies of biological characteristics of mesenchymal stem cells (MSCs) and regulatory factors that influenced the differentiation of MSCs have shown that the proportion of the natural differentiation from in vitro primarily cultured MSCs into hepatocytes was low, and to select a suitable inductor is important to enhance the differentiation of MSCs into hepatocytes.OBJECTIVE: To verify the feasibility of induced differentiation of rat bone marrow MSCs (BMSCs) into hepatocytes using the combination of hepatocyte growth factor (HGF), epidermal growth factor (EGF) and fibroblast growth factor (FGF-4).DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Experimental Center, Weifang Medical College in August 2007.MATERIALS: Totally 40 Sprague-Dawley rats were supplied by the Experimental Animal Center, Weifang Medical College.METHODS: Rat BMSCs were incubated by adherent method. BMSCs at passage 3 were assigned to 2 groups. BMSCs in the blank control group were treated with L-DMEM containing 10% fetal bovine serum. BMSCs in the combination group were treated with 10 μg/L FGF, 8 μg/L HGF and 8 μg/L EGF following above-mentioned procedures.MAIN OUTCOME MEASURES: Inverted microscope was used to observe the morphological changes in cells.Immunofluorescence method was used to observe the expression of alpha-fetoprotein (AFP) and albumin (ALB). PAS was employed to detect the expression of glycogen. Fox green intake experiment was conducted. Enzymology was utilized to test the contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP).RESULTS: BMSCs in the combination group presented polygonal, orbicular or round shape. BMSCs in the blank control group remained spindle. BMSCs in the combination group were positive for AFP and ALB at day 14 following culture, and a few PAS-positive and fox green-positive cells were found at day 7. Positive cells became more over time. Synthesis of ALT, AST and ALP was detected at day 14, reached a peak at 21 days, and then decreased. Above-described indexes were negative in the blank control group.CONCLUSION: After induced by the FGF, HGF and EGF, BMSCs have the ability to differentiate into hepatocytes in vitro.

OBJECTIVE To evaluate VITEK32 expert system(7.02) for detection and analysis of clinically important resistance phenotypes.METHODS A total of 508 known resistant phenotype clinial strains and 9 standards strains were tested by VITEK32 expert system(7.02) and antibiotic susceptibilities were determined by CLSI recommendation.RESULTS The correct phenotype was identified by the expert system in one or more choices for 312 from the 508(61.4%) isolates and standards.The resistant phenotypes for meticillin-susceptible,and resistant Staphylococcus spp,extended-spectrum ?-lactamases(ESBLs) producingEscherichia coli,Klebsiella spp,AmpC producing Enterobacter,cloacae,Serratia marcescens,and vancomycin-resistant Enterococcus faecalis were accurately identified by VITEK32 expert system,this expert system was not including ESBLs producing Proteus mirabilis.CONCLUSIONS VITEK32 expert system can be accurately identified most clinically important bacteria based on phenotype.The data of ESBLs Producing P.mirabilis should be included in the further work on expert system.