BackgroundMicroRNAs (miRNAs) play crucial roles in various physiological processes through post-transcriptional regulation of gene expressions and are involved in development, metabolism, and many other important molecular mechanisms and cellular processes. The Bombyx mori genome sequence provides opportunities for a thorough survey for miRNAs as well as comparative analyses with other sequenced insect species.Methodology/Principal FindingsWe identified 114 non-redundant conserved miRNAs and 148 novel putative miRNAs from the B. mori genome with an elaborate computational protocol. We also sequenced 6,720 clones from 14 developmental stage-specific small RNA libraries in which we identified 35 unique miRNAs containing 21 conserved miRNAs (including 17 predicted miRNAs) and 14 novel miRNAs (including 11 predicted novel miRNAs). Among the 114 conserved miRNAs, we found six pairs of clusters evolutionarily conserved cross insect lineages. Our observations on length heterogeneity at 5′ and/or 3′ ends of nine miRNAs between cloned and predicted sequences, and three mature forms deriving from the same arm of putative pre-miRNAs suggest a mechanism by which miRNAs gain new functions. Analyzing development-related miRNAs expression at 14 developmental stages based on clone-sampling and stem-loop RT PCR, we discovered an unusual abundance of 33 sequences representing 12 different miRNAs and sharply fluctuated expression of miRNAs at larva-molting stage. The potential functions of several stage-biased miRNAs were also analyzed in combination with predicted target genes and silkworm's phenotypic traits; our results indicated that miRNAs may play key regulatory roles in specific developmental stages in the silkworm, such as ecdysis.Conclusions/SignificanceTaking a combined approach, we identified 118 conserved miRNAs and 151 novel miRNA candidates from the B. mori genome sequence. Our expression analyses by sampling miRNAs and real-time PCR over multiple developmental stages allowed us to pinpoint molting stages as hotspots of miRNA expression both in sorts and quantities. Based on the analysis of target genes, we hypothesized that miRNAs regulate development through a particular emphasis on complex stages rather than general regulatory mechanisms.
BackgroundStunting and soil-transmitted helminth (STH) infections including ascariasis, trichuriasis and hookworm remain major public health problems in school-age pupils in developing countries. The objectives of this study were to determine the prevalence of stunting for children and its association with three major soil-transmitted helminths (STH) in rural areas of southern China. The study also aims to determine risk factors for stunting and to provide guidance on the prevention and control of stunting and STH infections for future studies in this field.ResultsA cross-sectional survey was carried out in the poor rural areas in Guangxi Autonomous Regional and Hainan Province where STH prevalence was higher between September and November 2009. Pupils were from 15 primary schools. All the school-age pupils aged between 9 and 12 years old (mean age 11.2 ± 3.2 years), from grades three to six took part in this study. Study contents include questionnaire surveys, physical examination and laboratory methods (stool checking for eggs of three major STH infections and haemoglobin determination was performed for the anaemia test). Finally 1031 school-age pupils took part in survey. The results showed that the overall prevalence of stunting (HAZ < 2SD) was 25.6%, based on the WHO Child Growth Standards (2007). Risk factors for stunting based on logistic regression analyses were: (1) STH moderate-to-heavy intensity infections (OR = 1.93;95%CI:1.19,3.11); (2) anaemia (OR = 3.26;95%CI: 2.02,5.27); (3) education level of mother (OR = 2.13; 95%CI: 1.39,3.25). The overall prevalence of major STH infections was 36.7%, STH moderate-to-heavy intensity infections was 16.7%. The overall prevalence of ascariasis, trichuriasis, hookworm and co-infection were 18.5%, 11.2%, 14.7% and 9.1% respectively. The prevalence of anaemic children (HB < 12 g/dl) was 13.1%.ConclusionThe present study showed that stunting was highly prevalent among the study population and STH infection is one of the important risk factors for stunting, with moderate-to-heavy intensity infections being the main predictor of stunting. Hence, additional interventions measures such as to promote de-worming treatment, to enhance health education and to improve hygiene and sanitation in order to reduce stunting in this population, are needed throughout the primary school age group.
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