ABSTRACT
OBJECTIVE To investigate the mechanism of catalpol affecting the differentiation of helper T cell 17 （Th17） by interfering the expressions of pyruvate kinase M2 （PKM2） and lactate dehydrogenase A （LDHA）. METHODS The naive CD4+ T cells were selected from the spleen of C57BL/6 mice， and were differentiated into Th17 cells by adding directional differentiation stimulants for 72 hours. At the same time， the cells were treated with 0 （directed control）， 20， 40 and 80 μg/mL catalpol. The flow cytometry was used to detect the proportion of Th17 cell differentiation in cells； the colorimetric method was adopted to detect the levels of pyruvate and lactate in cell culture supernatant； mRNA expressions of retinoid-related orphan nuclear receptor gamma t （RORγt）， PKM2 and LDHA were detected by qRT-PCR method； Western blot was used to detect the expression levels of PKM2， LDHA， signal transducer and activator of transcription 3 （STAT3）， and phosphorylated STAT3 （p-STAT3） proteins in cells. RESULTS Compared with the directed control group， after 72 hours of treatment with 20， 40， 80 μg/mL catalpol， the differentiation ratio of Th17 cells were decreased by 6.74%， 8.41%， 9.24%， and the levels of pyruvate and lactate in the cell culture supernatant， the mRNA expressions of PKM2， LDHA and RORγt as well as the protein expressions of PKM2 and LDHA and the phosphorylation of STAT3 were significantly reduced （P＜0.05）. CONCLUSIONS Catalpol can reduce the glycolysis level by down-regulating the expressions of PKM2 and LDHA， thereby inhibiting the differentiation of Th17 cells.
ABSTRACT
Objective To investigate the correlation between atlE gene and biofilm formation of Staphylococcus epidermidis . Methods 64 strains of clinically isolated Staphylococcus epidermidis in our hospital from June 2015 to June 2016 were collected . The biofilm formation test was used to detect bacterial biofilm .PCR was use to amplify atlE gene .Then the correlation between the atlE gene with biofilm formation was analyzed .Results 24 strains of biofilm positive bacterium were detected ,the detection rate was 37 .5% ;31 strains of atlE gene was detected ,the detection rate was 48 .4% ;atlE gene was significantly correlated to biofilm formation(P＜0 .05) .Conclusion Staphylococcus epidermidis has the ability to form biofilm ;atlE gene has a relation with biofilm formation of Staphylococcus epidermidis .
ABSTRACT
Objective To investigate the formation of biofilm in clinical isolates of Staphylococcus epidermidis ,and to analyse the correlation between biofilm formation and antibacterial resistance of Staphylococcus epidermidis .Methods A total of 62 strains of Staphylococcus epidermidis isolated from blood specimens of inpatients with bloodstream infection ,from January 2014 to February 2015 ,were collected .The biofilm formation of Staphylococcus epidermidis was detected by using the semi‐quantitative adherence as‐say and polymerase chain reaction(PCR) amplification experiment .The antibacterial susceptibility test was carried out according to K‐B method .Results The positive rate of biofilm formation detected by using the semi‐quantitative adherence assay and PCR for icaA gene were 37 .1% (23 strains) and 43 .5% (27 strains) respectively ,and there was no statistically significant difference(P>0 .05) .There were 14 positive strains detected by both methods .The resistance rates of strains producing biofilm to antibacterial a‐gents were generally higher than those of non‐producing biofilm strains ,and there were statistically significant differences in resist‐ance rates of strains to gentamicin ,penicillin ,oxacillin ,levofloxacin and cefoxitin(P<0 .05) .All bacteria were sensitive to vancomy‐cin ,linezolid and quinupristin/dalfopristin .Conclusion There is no significant difference between the two methods in detecing bio‐film formation .The resistance rates of strains producing biofilm to antibacterial agents were generally higher than those of non‐pro‐ducing biofilm strains .
ABSTRACT
Objective To investigate the infection status and drug suscepetibility of mycoplasma from 6 573 patients with non-gonococcal urethritis ,and to provide the scientific bases for the clinical application of antibiotics .Methods Mycoplasma detection kit was used to detect ureaplasma urealyticum (Uu) and mycoplasma hominis(Mh) and the drug susceptibility .All the patients were divided into two groups :Chinese group and foreigner group .Results Among 5 675 Chinese patients ,2 985 patients were infected by mycoplasma(52 .6% ) .The infection rate of Uu was 2 312(40 .7% ) .35 .2% patients were male ,and 61 .4% patients were female .In 898 foreign patients ,440 patients were infected by mycoplasma(49 .0% ) .The infection rate of Uu was 327(36 .4% ) .32 .2% pa-tients were male ,and 59 .5% patients were female .In Chinese patients infected by Uu ,the susceptibility rates to MIN ,DOX ,JOS and CLA were 96 .7% ,96 .2% ,93 .7% ,89 .7% ,respectively .In foreign patients ,the susceptibility rates to MIN ,DOX ,JOS ,and CLA were 98 .9% ,98 .4% ,95 .8% ,92 .1% .Conclusion The mycoplasma infection rate of Chinese patients is higher than foreign patients .In both groups ,Uu infection is the main type .Female patients are more than male patients .The drug sensitivity rate in for-eign group is higher than that in Chinese group .mycoplasma are sensitivity to MIN ,DOX ,JOS .
ABSTRACT
Objective To elucidate the expression of gankyrin in human gastric cancer cells and it's role in nimesulide induced apoptosis. Methods Four human gastric cancer cell lines including MKN28 (well differentiated), AGS (poorly differentiated), MKN45 (poorly differentiated), and SGC7901(moderately differentiated) were cultured and treated with nimesulide. Nimesulide induced growth inhibition and apoptosis of the cells were detected by methyl thiazolyl tetrazolium assay, and confirmed by flow cytometry. The expressions of gankyrin gene and protein were further assessed by real-time PCR and Western blotting. Results Gankyrin mRNA and protein were detected in all four human gastric cancer cell lines. The proliferations of AGS and SGC7901 cell lines were significantly suppressed by nimesulide in a time-dose dependent manner. When treated with 400 μmol/L of nimesulide for 48 hours, the significant apoptosis was found in AGS cells (23.30%±2.50%) and SGC7901 cells (16.80%±1.55% ) in comparison with controls (0.57%±0.19% and 0.88%± 0.17%, respectively, all P values <0.01). Apoptosis of AGS cells induced by nimesulide was accompanied by a considerably decreased gankyrin expression that was more significant at 24 hours (0.0035±0.0014) and 36 hours (0.0980±0.0160) in comparison with controls (0.4690±0.1190, all P values<0.01). Conclusion Gankyrin expresses in human gastric cancer cell lines and may be involved in nimesulide induced apoptosis of AGS cells.