Effectiveness and safety of oseltamivir for treating influenza: an updated meta-analysis of clinical trials

<div><p></p><p><i>Background:</i> Oseltamivir has been widely used to treat patients with influenza; however, its effects have been debated. Recently, a new meta-analysis on the controversial topic of oseltamivir’s effectiveness found that the drug reduces the duration of influenza symptoms and the risk of hospitalization, while increasing the risk of nausea and vomiting. Unfortunately, the analysis did not include articles published in Chinese. Thus, we performed an updated meta-analysis by adding more studies in China. <i>Methods:</i> The terms ‘oseltamivir,’ ‘influenza,’ and ‘effect’ were used to search for publications in both English and Chinese. Only controlled clinical trials were included. We used the weighted mean difference (WMD) or relative risk (RR), together with the 95% confidence interval (95% CI), to estimate the effects of oseltamivir. <i>Results:</i> A total of 12 studies including 107 712 patients were eligible for analysis. Oseltamivir significantly reduced the duration of fever (WMD, –20.48; 95% CI, –28.43, –12.53) and influenza-like symptoms (WMD, –19.39; 95% CI, –32.94, –5.84). The rates of hospitalization (RR, 0.79; 95% CI, 0.68, 0.90), antibiotics usage (RR, 0.56; 95% CI, 0.42, 0.74), otitis media (RR, 0.78; 95% CI, 0.65, 0.93), and nonspecific complications (RR, 0.58; 95% CI, 0.35, 0.95) were significantly decreased among patients taking oseltamivir. No significant difference was observed with respect to the risk of adverse reactions. <i>Conclusion:</i> Oseltamivir can effectively alleviate the symptoms of influenza and reduce hospitalization, antibiotic usage, and the risk of otitis media without significantly increasing the rate of adverse drug reactions.</p></div

Label-Free MicroRNA Profiling Not Biased by 3′ End 2′-O-Methylation

Accurate quantification of miRNA expression level is essential to the study of its biology, and many cutting-edge technologies have been developed to accommodate this need. Yet most of them were designed primarily for the “regular” RNAs such as animal miRNAs and may overlook the fact that plant miRNAs and many other small noncoding RNAs are 2′-O-methylated at the 3′ end nucleotide. According to our experimental data and previous reports, this structural variation is detrimental to the effectiveness of the commonly used enzymatic labeling methods, leading to strongly biased results (∼24-fold difference). Herein, we demonstrate that our Stacking-Hybridized Universal Tag (SHUT) microarray assay is well suited for unbiased profiling of both normal and methylated small RNA species. The detected signals of small RNAs with 2′-hydroxyl and 2′-<i>O</i>-methyl 3′ ends are highly consistent (no significant difference at α = 0.01 level). For specificity, the presented method edges over others by its unique ability to discriminate single-base difference at or near the 5′ end. Notably, as compared to many delicate techniques, this enzyme-free and label-free approach requires much less reagent and manipulation, benefiting the SHUT-based applications with more efficient workflow and highly reproducible results

Effect of SF on the levels of total and phosphorylated proteins involved in the G2/M cell cycle arrest.

<p>(A) The protein levels of KLF4, p21, p53, cyclin B1, cyclin D1, cdc2, and cdc25C and phosphorylation of cdc2 (Tyr 15) and cdc25C (Ser 216) were evaluated by immunoblots in LCL, pre-B ALL, and T-ALL cell lines incubated with 7.5 µM SF or vehicle for 24 hours. β-actin was used as a loading control. The data are representative of three independent experiments. (B) Detection of phospho-H2AX (pH2AX) in nuclei by flow cytometry in Nalm-6 and Jurkat cells treated with 7.5 µM of SF or Etoposide (ETO) for 24 hours. The data are representative of three independent experiments. (C) Diagram depicting the role of SF in the G2/M cell cycle arrest of ALL cells.</p

SF inhibits the AKT/mTOR survival pathway in leukemic cells.

<p>The protein levels of phosphorylated and total AKT and mTOR were examined by immunoblotting. LCL, pre-B ALL, and T-ALL cell lines were incubated with 7.5 µM SF or vehicle for 24 hours. β-actin was used as a loading control. The data are representative of three independent experiments.</p

SF induces apoptosis selectively in ALL cell lines.

<p>(A) Apoptosis was evaluated by annexin-V and 7-AAD staining of LCL, Nalm-6, Jurkat and KOPTK1 cells cultured in the absence or presence of 7.5 µM SF. The data are representative of three independent experiments. (B) The percentages of annexin-V-positive cells were determined for each cell line cultured in the presence or absence of SF. (C) SF activates the proteolytic cascade of caspases and PARP in leukemic cells. LCL cells (control), pre-B ALL cells (Nalm-6, REH, and RS-4), and T-ALL cells (Jurkat, RPMI, DND41, and KOPTK1) were incubated with 7.5 µM SF for 24 hours and analyzed by immunoblotting. The arrows indicate the cleaved forms of the caspases and PARP. β-actin was used as a loading control. The data represent the mean and standard deviation (n = 3). *** <i>P</i><0.001 (two-tailed Student’s <i>t</i>-test).</p

<i>In vivo</i> anti-leukemic effect of SF in ALL xenograft models.

<p>(A) Nalm-6 cells were transduced with the FFluc retrovirus and injected intravenously into NOD/SCID mice. Twenty-four hours later, the mice were treated with SF or vehicle (2 mg i.p. daily) and monitored weekly for the distribution of leukemic cells by bioluminescence imaging (BLI). (B) The Nalm-6-FFluc cells were mixed with a high-protein matrigel and injected into the flank of NOD/SCID mice. One week later, tumor establishment was confirmed by BLI; then, the mice were treated by oral gavage (2 mg/gave, twice daily) for 7 days. The total counts were determined for the control- and SF-treated mice. The data are representative of three independent experiments. The statistical significance was calculated using the two-tailed Student’s <i>t</i>-test.</p

In vitro splicing assay of the c.1439+1G>C mutation.

<p>(A) The splice region (c.1439+1, intron 9) of <i>SCNN1A</i> gene was amplified and products were ligated into the pcDNA3.1/Myc-His B vector. (B) RT-PCR of HEK293 cells transfected with either wild-type or mutant <i>SCNN1A.</i> Minigenes showed that the mutation c.1439+1G>C was sufficient to produce a longer band. (C) Lane 1: Empty pcDNA3.1 vector; Lane 2: wild-type <i>SCNN1A</i> (256 bp); Lane 3: c.1439+1G>C mutant (361 bp); Lanes 4, 5, and 6: <i>GAPDH</i> used as control (245 bp).</p

Three novel mutations were identified in the <i>SCNN1A</i> gene.

<p>(A) Sequences showing a compound heterozygous mutations (c.1311delG in exon 8 c.1439+1G>C in intron 9) in PHA1 patient of case 1. (B) Sequences showing a homozygous mutation (c.814_815insG in exon 4) in PHA1 patient of case 2.</p

Bivariate Results^a for Density of and Distance to Food Outlets between Tribal Areas^b and Non-Tribal Areas^c.

<p>Bivariate Results<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161132#t003fn001" target="_blank">^a</a> for Density of and Distance to Food Outlets between Tribal Areas<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161132#t003fn002" target="_blank">^b</a> and Non-Tribal Areas<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161132#t003fn003" target="_blank">^c</a>.</p

Demographic Characteristics of Participants Providing Qualitative Interviews.

<p>Demographic Characteristics of Participants Providing Qualitative Interviews.</p