Objective To explore the effects of interleukin- 1?(IL-1?)on the biological activity of human periodontal ligament(PDL) cells in vitro. Methods Human PDL cells were cultured in DMEM medium containing IL-1?(0.1,0.5,1,5 and 10ng/ml) for 1,2,3,4 and 5 days, respectively. The proliferation of PDL cells was measured by MTT assay at the first, second, third, fourth and fifth days after IL-1? treatment, respectively. Fibronectin level in the medium was determined by ELISA at the fourth days after 10ng/ml IL-1? treatment, and alkaline phosphatase(ALP) activity was measured by enzyme kinetic method. Results IL-1? inhibited the growth of human PDL cells in a dose-dependent manner, and its lowest effective concentration was 1ng/ml(P

Objective To investigate the antibiotic resistance and molecular epidemiology character of ?-lactamase producers among clinical isolates of Citrobacter freundii.Methods Four strains of Citrobacter freundii were detected by K-B susceptibility method and three-dimensional test.The ?-lactamase gene was identified by polymerase chain reaction(PCR) and sequencing.Results The No.29 strain of Citrobacter friundii was multiple-drug-resistant and proved to produce CMY,DHA type AmpC,TEM and SHV type of extended-spectrum ?-lactamases(ESBLs).The sequencing of corresponding DNA revealed 97% identities of the CMY-2 amino acid sequence.It was a new type of cephalosporinase.Conclusion A novel strain of Citrobacter friundii with CMY type AmpC,DHA type AmpC,TEM and SHV type ESBLs has been found in southern China and it is multiple-drug-resistant.

OBJECTIVE To investigate the phenotypical and genotypical characteristics of the ?-lactamase in the 302 strains of Gram-negative bacilli,and assess the characteristics of the drug resistance.METHODS The phenotypes of ?-lactamase were detected in the Gram-negative bacilli by disc diffusion screen test,which were further ascertained by disc diffusion confirmatory test,a cefoxitin three dimensional test,and metallo ?-lactamases double-disc synergy test.The ?-lactamases were detected by multiple PCR amplification,then DNA product was sequenced.Antibiotics susceptibility was detected by K-B method.RESULTS Among the clinical Gram-negative isolates,a total of 26.49%(80/302)strains′ ?-lactamases phenotypes were positive,ESBLs,ESBLs combined with AmpC enzyme,ESBLs combined with AmpC enzyme and MBL,and AmpC enzyme producing strains were found in 16.56%,6.62%,1.66% and 0.66%,respectively.A total of 24.83% strains′?-lactamases phenotypes were positive,strains produced ESBLs,ESBLs combined with AmpC enzyme,ESBLs combined with AmpC enzyme and MBL,and AmpC enzyme producing strains were found in 17.55%,6.95%,0.33%,and 0.33%,respectively.Drug-sensitivity test showed multi-resistant Gram-negative isolates were more sensitive to amikacin(18.75%),and heavier resistant to ampicillin(97.50%).CONCLUSIONS ?-Lactamase producing Gram-negative bacilli are resistant to multi-antibiotics.

Objective To investigate the distribution of the CTX-M- extended spectrum beta-lactamase (ESBLs) producing Esche- richia coli(ECO) and the molecular mechanism of dissemination. Methods To analyze the drug resistance of the 43 isolates, Kirby-Bauer susceptibility method was used. Multiple polymeraso chain reaction (PCR) was used to amplify the gene of ESBLs, AmpC, full length of blaCTX-M-like gene, insertion sequence (IS) ISEcp1B, IS903 , IS26 and integron I. NEST-PCR was used to detect if the beta-lactamase gene lo- cated in the integron I. The product of full length of bla-CTX-M like gone amplified by PCR was sequenced. Results Susceptibility test showed the resistance from high to low in turn was Ampicillin (97.68%), Coftriaxone (67.44 % ), piperacillin(65.12 % ), Cefotaxime (62.79 % ) ,Coftasidime(58.14% ), Cofasolin(55.81% ), Cofepime (53.49%), Cefexitin(51.16%), ciprofloxacin (44. 19% ), Aztreo- nam(41.86% ), Cefoperasone/Sulbactam ( 20.93% ), Amikacin (0% ), Imipenem (0% ), respectively. ECO was susceptive to Imipenem. CTX-M-G1 was found in 25 strains of ECO , TEM, SHV, CTX-M-G1, ISEcp1B, and integron I were found in the nine isolates. IS903 were found in ECO 3 and 5, and IS26 was found in ECO 3. In ECO 3 and 5, blaCTX-M-like was flanked upstream by ISEcp1B element that provided -35 and -10 promoter sequences and a right inverted repeat (IRR) recognized by transposase, downstream by IS903 provided an inverted re- peat, ISEcp1 B and IS903 composed the complex transpeson. Conclusion ISEcplB may drive the expression and dissemination of blaCTX-M-like gene at a high level.

Objective To analyze the infection status and variation tendency of chlamydia trachomatis (CT) ,Neisseria gonorrhoe-ae(NG) and ureaplasma urealyticum(UU) in female genital tract in northeast Sichuan province during 2005 -2012 to provide the laboratory basis for their diagnosis and treatment Methods The real-time fluorescent quantitative PCR (RT-PCR) was applied to detect the CT DNA ,NG DNA and UU DNA in 1 386 samples from female genital tract and the detection results were performed the statistical analysis .Results The total positive rate of these 3 kinds of pathogens was 62 .8% (871/1 386) .Among the simple in-fection ,UU had the highest positive rate(48 .0% ,665/1 386);the positive rates of CT and NG were only 2 .2% .In the mixed infec-tion ,the positive rate of CT + UU was highest(6 .5% ,90/1 386) ,while which of UU + NG ,CT + NG and CT+ NG+ UU was 2 .5% (35/1 386) ,0 .4% (5/1 386) and 1 .1% (15/1 386) respectively .In different age groups ,the positive rate in the age <20 years old group was 49 .3% ,while which in the age >20 years old groups were all more than 60% .The positive rate of the CT ,NG and UU pathogens in females was in continuous high level during 2005 -2012 ,and which totally showed an increasing tendency . Conclusion CT and UU are the main pathogens in female genital tract infection in this region ,and the positive rate of genital tract infection in females aged more than 20 years is higher ,the infection rate of these 3 kinds of pathogens demonstrates the increasing trend year by year ,so more attention should be paid to the detection of CT and UU in this group for guiding the clinicians to con-duct the diagnosis and treatment .

Objective To investigate the distribution of high pathogenicity island(HPI)in multiple-drug-resistance gram-negative bacilli and analyze the protein sequence.Methods To amplify thefyuA-irp2 gene cluster of the 84 isolates by multiple polymerase chain reaction(PCR),the product was subsequently sequenced.Results The positive rate ofirpl,irp2,irp3,irp4 and fyuA was 40.48％,41,67％,5.95％,O％and 16.67％,respectively.Theamino sequence offyuA comefromEC06748,Kp7151 and PAE7 was usedto compare with AL590842,there are 100％identities.Amino sequence ofirp2 come from Kp49 and Kp51 have 99％identities with AAA27636.1,but amino sequence of irp2 come from EC04 and EC07 only have 90％identities with 1176840.The GenBank accession number is FJ211852 and FJ211851.Amino sequence ofirpl come fromKp 10,Kp49 and Kp51 have 99％identities with AL590842。and amino sequence ofirp3 come from EC03,Kp51,Kp10 and Kp49 have 97％identities with CAA73128.There are the same mutation among the same species,and different mutation among different species.Conclusion There was different extant mutant lost in thefy～t-i,v2 gene cluster in multiple-drug-resistanee gram-negative bacilli.

Objective:To investigate the mechanism of Yunshi Ganmao Heji against respiratory syncytial virus (RSV) infection based on network pharmacology and in vivo experiments. Methods:Network pharmacological prediction: Several databases including TCMSP and GeneCards were used to predict the active ingredients and targets of Yunshi Ganmao Heji in the intervention of RSV infection. Cytoscape 3.2.1 software was used to construct the traditional Chinese medicine component-disease target network diagram. The interactions between proteins were analyzed by STRING database. GO functional enrichment analysis and KEGG pathway enrichment analysis were performed using Metascape database. Molecular docking technology was used to verify the results of network pharmacology. Experimental verification of Yunshi Ganmao Heji for the intervention of RSV infection: A mouse model of RSV infection was established through intranasal infection. After the administration of Yunshi Ganmao Heji, blood routine test results, lung indexes and pathological changes in lung tissue were analyzed. Peripheral blood T cell subsets were detected by flow cytometry. The levels of TNF-α, IL-6 and IL-1β in serum were detected by ELISA. RT-PCR was used to detect the relative expression of TLR4, NF-κB and RSV-N gene at mRNA level in lung tissues.Results:A total of 41 active ingredients of Yunshi Ganmao Heji and 111 drug targets for RSV infection were obtained. Besides, 167 signaling pathways mainly including PI3K/AKT, MAPK and Toll-like receptor signaling pathways were obtained. Molecular docking results showed that the binding energies of luteotin, kaempferol and quercetin, three active ingredients of Yunshi Ganmao Heji, with RSV-G, RSV-F, PI3K, AKT1 and Bcl-2 were less than 0 kcal/mol. In vivo experiment results showed that compared with RSV group, the counts of white blood cells and lymphocytes increased and the lung index decreased in high-dose Yunshi Ganmao Heji group, with statistically significant difference ( P<0.05). HE staining showed pulmonary hyperplasia, thickened alveolar wall and inflammatory cell infiltration in interstitium in RSV group. Alveoli in ribavirin group as well as low-dose, medium-dose and high-dose Yunshi Ganmao Heji groups tended to be of uniform size, and the alveolar walls was roughly uniform in thickness. Compared with RSV group, the low-dose, medium-dose and high-dose Yunshi Ganmao Heji groups showed significantly increased numbers of CD3 +, CD4 + and CD8 + T lymphocytes, decreased CD4 + /CD8 + T cell ratio, lower levels of TNF-α, IL-6, IL-1β in serum, and reduced viral load and inhibited expression of TLR4 and NF-κB at mRNA level in lung tissues ( P<0.05). Conclusions:Yunshi Ganmao Heji can regulate RSV infection by targeting multiple targets and pathways with several active ingredients. Its main functions are to alleviate pathological injury in lung tissues and reduce inflammatory response, and the possible mechanism underlying the antiviral functions may be related to its inhibitory effect on the activation of TLR4/NF-κB pathway.