Inhibitory effect of siRNA-mediated MSK1-silencing on the proliferation of CNE2 cells and its mechanisms / 医学研究生学报

Objective Abnormal activation of mitogen-and stress-activated kinase (MSK1) plays an important role in the development of various cancers.This study was to explore the effect of small interfering RNA (siRNA)-mediated MSK1-silencing on the proliferation of human nasopharyngeal carcinoma (NPC) cells and its underlying mechanism.Methods The siRNA vector targeting MSK1 was constructed and transfected into CNE2 cells, and the NPC cell line stably expressing MSK1 was established.Then the cells were divided into a blank control (without transfection of the plasmid), a negative control (with stable transfection of the negative control plasmid), and an experimental group (with stable transfection of the positive recombinant plasmid).The expressions of MSK1 mRNA and protein were detected by real-time quantitative PCR and Western blot, respectively, the proliferation of the cells determined by CCK-8 and colony formation assays, the cell cycles analyzed by flow cytometry, the level of histone H3 phosphorylation at Ser10 examined by Western blot, and The transcriptional activity and expression of the c-jun protein measured by dual-luciferase reporter gene assay and Western blot.Results Compared with the blank control, the inhibition rates of cell proliferation at 48, 72 and 96 hours were significantly reduced in the experimental group (P<0.05), and so were the colony formation ability of the cells (P<0.01) and the expression and transcriptional activity of the c-jun protein (P<0.05).In comparison with the negative control, the experimental group showed significant decreases in the rate of cell growth after 24 hours, the inhibition rates of cell proliferation at 48, 72 and 96 hours (P<0.05), the number of formed colonies ([221.00±20.08] vs [99.67±15.57] / 300 cells, P<0.01), the proportion of S-phase cells (P<0.01), and the expression of the c-jun protein in the CNE2 cells ([100.00±0.00] vs [48.77±10.71] %, P<0.05), but a remarkable increase in the percentage of G0/G1-phase cells (P<0.01).Furthermore, histone H3 phosphorylation at Ser10 was markedly reduced (P<0.01) but no significant change was observed in the expression of the total c-jun protein in the experimental group.Conclusion Knockdown of MSK1 using siRNA can significantly inhibit the growth and proliferation of CNE2 cells, which may be closely related to the decreased phosphorylation of histone H3 and subsequently down-regulated transcriptional activity of c-jun.

Value of procalcitonin and lipopolysaccharide in identifying pathogens and evalu-ating therapeutic efficacy of hospital-acquired pneumonia / 中国感染控制杂志

Objective To explore the value of procalcitonin (PCT)and lipopolysaccharide (LPS)in identifying pathogens and evaluating therapeutic efficacy of hospital-acquired pneumonia (HAP).Methods A total of 110 HAP patients were enrolled in a prospective study,patients were divided into gram-negative bacterial infected HAP group (G- infected group,n=50),gram-positive bacterial infected HAP group (G+ infected group,n=30),and control group (nontypical pathogen or virus infected group,n =30).Serum levels of PCT,LPS and C-reactive protein (CRP)of patients were dynamically detected,receiver operating characteristic (ROC)curve and area under the curve (AUC)were adopted to assess the value of PCT and LPS in predicting pathogenic bacteria causing HAP. Results PCT and LPS levels of G - infected group were (3.43 ±1 .15)ng/mL and (0.20 ±0.08)EU/mL respec-tively,which were higher than G+ infected group ([0.42±0.12]ng/mL and [0.05±0.02]EU/mL respectively)and control group([0.14±0.08]ng/mL and [0.02 ±0.01 ]EU/mL respectively)(all P <0.05 ).Levels of PCT and CRP of G- infected group before and after therapy were both significantly different ([3.43±1 .15]ng/mL vs [0.63 ±0.22]ng/mL,[47.26±30.35]mg/L vs [9.21 ±6.54]mg/L,respectively)(both P <0.01).The levels of PCT, LPS,and CRP in moderate and severe patients were all significantly higher than mild patients ([5.43±1 .05]ng/mL vs [0.72±0.32]ng/mL,[0.33±0.07]EU/mL vs [0.09 ±0.04]EU/mL,[57.46 ±20.15 ]mg/L vs [8.25 ± 5.24]mg/L,respectively)(all P <0.05).Sensitivity and specificity of combined detection of PCT and LPS in dif-ferentiating gram-negative bacteria infected VAP from gram-positive bacteria infected VAP were 95.83% and 96.15% respectively,AUC was 0.95.Conclusion PCT and LPS have certain value in identifying pathogens of HAP,combined detection of PCT and LPS can increase specificity in identifying HAP type,and assess the efficacy of antimicrobial therapy in accordance with the dynamic change.

Study on the relationship between polymorphism sites of IRF5 and SLE patients in Shandong Han population / 中华微生物学和免疫学杂志

Objective To investigate two single nucleotide polymorphism sites of IRF5 and to de-tect their relationship with SLE in a population from Shandong province. Methods The polymorphisms (rs2004640 G/T,rs10954213 G/A) were detected with PCR-RFLP in 92 eases of SLE and 88 healthy con-trols. The genotype and allele frequencies were calculated and analyzed. Results The genotype frequencies Of GG, GT and TT in rs2004640 site in SLE were 0. 198, 0.521 and 0.281, respectively. The difference was significant between SLE and centrol (X2 = 8.73, P < 0.05). The genotype frequencies of GG, GA and AA in rs10954213 site in SLE were 0. 318, 0. 409 and 0.273, respectively. The differenee was significant between SLE and control (X2 = 6. 36, P < 0. 05). Conclusion The polymorphism of rs2004640, rs10954213 in IRF5 may be associated with SLE in the population of Han nationality from Shandong province of China.

mRNA expression of interferon regulatory factor 5 and Toll like receptor-9 in patients with systemic lupus erythematosus / 中华皮肤科杂志

h may play a role in the pathogenesis of SLE.