Hsa_circ_0009057 can be a novel potential diagnostic biomarker and a therapeutic target for premature brain injury.

Circular RNAs (CircRNAs) represent a novel class of noncoding RNAs, and are widely expressed in brain tissues, indicating their great potential as novel biomarkers for the diagnosis of neuropsychiatric disorders. However, the expression profiles of circRNAs for infants with premature brain injury (PBI) have not been fully explored. This study assessed the differentially expressed circRNAs in plasma of infants with and without PBI using the human circRNA microarray analysis. Hsa_circ_0009057, a significantly up-regulated circRNA in infants with PBI, was selected for further study. The microRNA (miRNA)-recognition element prediction analysis and construction of the circRNA-miRNA-mRNA network showed that hsa_circ_0009057 was associated with several miRNAs and mRNAs, which played important roles in the regulation of neuropsychiatric disorders. The results of the receiver operating characteristic curve analysis showed that hsa_circ_0009057 had high degrees of specificity and sensitivity. The results also indicated that hsa_circ_0009057 could be served as a novel potential diagnostic biomarker and a therapeutic target for PBI.

Long noncoding RNAs interact with mRNAs: A new perspective on the mechanism of premature brain injury.

The molecular mechanism of premature brain injury induced by inflammation is not fully understood. Long noncoding RNAs (lncRNAs) have been reported to play crucial roles in neurological disorders including brain injury. However, little is known about the regulatory function of lncRNAs in the premature brain. This study investigates differentially expressed lncRNAs and mRNAs as well as their interactions in the premature brain. Lipopolysaccharides (LPS) were used to induce inflammation in premature rodent models. Brain histology was observed via hematoxylin and eosin (HE) staining and CD68 immunostaining. Arraystar microarry was designed for the profiling of differentially expressed lncRNAs and mRNAs in 4 LPS induced premature brains (L group), 4 full-term control brains (C group) and 3 premature brains were not induced by LPS (P group). Bioinformatic analysis was applied to reveal the functions and co-expression relationship of lncRNAs and mRNAs. Three lncRNAs and 2 mRNAs were selected for validation applying quantitative real time polymerase chain reaction (qRT-PCR). This study demonstrates dysregulated lncRNA and mRNA profiles in the premature brains upon inflammatory insult, thus revealing a novel mechanism of premature brain development from a new perspective of the lncRNAs and mRNA coexpression network and providing important insights into the therapy of premature brain injury.

Adoptive transfer of T regulatory cells inhibits lipopolysaccharide-induced inflammation in fetal brain tissue in a late-pregnancy preterm birth mouse model.

To evaluate the effect of regulatory T cells (Tregs) on the inflammation resulting from lipopolysaccharide (LPS) challenge in prenatal brain tissue, Tregs isolated from pregnant mice were transferred into model mice, and the expression levels of fork head family transcription factor (Foxp3), interleukin-6 (IL-6), CD68 (a marker of microglia), and toll-like receptor 4 (TLR-4) were assessed in the fetal brain tissue. Foxp3, IL-6, and TLR-4 expression were detected by polymerase chain reaction and Western blot; CD68 expression level was detected using immunochemical analysis. Foxp3, IL-6, TLR-4, and CD68 expressions in fetal brain were significantly induced by maternal LPS administration, and the increased expression levels were markedly reduced by adoptive transfer of Tregs. Maternal LPS exposure significantly induced inflammation in perinatal brain tissue, and Tregs negatively regulated this LPS-induced inflammation.

[Screening and validation of long non-coding RNAs in brain tissue of inflammation-induced preterm mice].

OBJECTIVE: To investigate the association between long non-coding RNAs (lncRNAs) and brain injury in inflammation-induced preterm mice, and to provide a reference for the prevention and treatment of brain injury. METHODS: An intraperitoneal injection of lipopolysaccharide in pregnant mice was performed to establish a model of inflammation-induced preterm mice with brain injury (preterm group). The full-term mice delivered by normal pregnant mice were used as controls (full-term group). The lncRNA chip assay was used to screen out the lncRNAs associated with brain injury in preterm mice. Quantitative real-time PCR was used to validate the lncRNAs identified by the above method. RESULTS: The preterm and full-term groups showed significant differences in the expression of 1 978 lncRNAs (P<0.05), consisting of 786 up-regulated lncRNAs and 1 192 down-regulated lncRNAs, and 29 lncRNAs were 1.5 or more times differentially expressed between the two groups. A further analysis was performed for the 10 most differentially expressed lncRNAs, and the results showed that these lncRNAs were involved in the biological processes including transcription, signal transduction, apoptosis, cell cycle, and inflammatory response, as well as G protein-coupled receptor signaling pathway and neuropeptide signaling pathway. Real-time PCR was performed to validate the expression of two lncRNAs in brain tissue in the preterm and full-term groups, and the results were consistent with those of the chip assay. CONCLUSIONS: The expression profiles of lncRNAs in brain tissue change significantly in inflammation-induced preterm mice, and the G protein-coupled receptor signaling pathway may be involved in the pathogenesis of preterm brain injury.

Epidemiologic and etiologic characteristics of hand, foot, and mouth disease in Chongqing, China between 2010 and 2013.

Hand, foot, and mouth disease (HFMD) has become very common in children, with widespread occurrence across China. The aim of this study was to investigate the epidemiologic and etiologic characteristics of HFMD, including etiologic variations in Chongqing, China. An epidemiologic investigation was based on 3,472 patients who presented with HFMD manifestations and were admitted at the Children's Hospital of Chongqing Medical University between 2010 and 2013. Fecal specimens from 830 patients were analyzed by nested RT-PCR to identify the enterovirus pathogens, and the molecular characterization of HFMD was illustrated by phylogenetic tree analysis. The results of this study indicate that the peak of the HFMD epidemic in Chongqing between 2010 and 2013 occurred between April and July each year. The median age of onset was 2.24 years old, and children under the age of five accounted for 96.4% of all the HFMD cases; the male-to-female ratio was 1.89:1. Enterovirus 71 accounted for a major proportion of the isolated strains every year, including the majority (74%) of severe cases. However, the proportion of Coxsackie A (CV-A) 6 infections increased from 2.11% in 2010 to 16.36% in 2013, while the proportion of CV-A16 infections decreased from 31.23% in 2010 to 4.67% in 2013. Molecular epidemiologic study showed that all enterovirus 71 strains belonged to subgenotype C4a, whereas all CV-A16 strains belonged to genotype B1, including subgenotype B1a and subgenotype B1b.

[Acute toxicity of LDS, a molluscicide, to non-target organisms].

OBJECTIVE: To evaluate the acute toxicity of 10% LDS, a new molluscicide, to non-target organisms. METHODS: Based on "Chemical pesticide environmental safety test evaluation standard", an acute toxicity test was carried out with Coturnix coturnix japonica (quail), Apis mellifera L (bee), Bombyx mori (silkworm), and Brachydonio rerio (zebra fish) , and the skin stimulus test was also performed with guinea pig. RESULTS: The quails had no toxic symptoms while the maximum poisoning concentration of LDS was 200 mg/kg (no toxicity). LC50 of bees was 2.68 x 10³ mg/L (low toxicity). After 96 hours, no silkworms died in each group of different concentrations of LDS while the most concentration was 6.00 x 10² mg/kg, but there were some toxic symptoms such as inappetence and inactive in the high concentration group as compared to the blank control group (low toxic). LC50 (96 h) of zebra fish was 6.16 mg/L (medium toxicity). CONCLUSIONS: LDS has no toxicity to Coturnix coturnix japonica, low toxicity to Apis mellifera L and Bombyx mori, and medium toxicity to Brachydonio rerio. Compared with niclosamide ethanolamine salt, the toxicity to the fish is lower, and therefore, it is more suitable for the field application.

[Evaluation on molluscicidal effect of chlorosalicylicamide on Oncomelania hupensis in schistosomiaisis endemic areas of eight provinces in China].

OBJECTIVE: To evaluate the effects of a novel molluscicide, the salt quinoid-2', 5-dichloro-4'-nitrosalicylanilide from niclosamide (LDS), with 10% wettable powder, in main schistosomiasis epidemic areas of China, including Hunan, Jiangxi, Hubei, Anhui, Jiangsu, Sichuan, Yunnan and Zhejiang Province. METHODS: In the immersion test, 6 effective concentrations of 10% LDS were tested respectively: 0.1, 0.2, 0.4, 0.6, 0.8, 1.0 g/m3 in the field; at the same time, 50% wettable powder of niclosamide ethanolamine salt (WPN) with effective concentrations of 1.0 g/m was used as the molluscicide control, and the fresh water as the blank control, then the mortality rates of 0. hupensis snails were recorded at 24 h, 48 h and 72 h after the immersion. In the spraying test and powder-spraying test, 5 effective dosages of 10% LDS were tested respectively: 0.2, 0.4, 0.6, 0.8, 1.0 g/m2, while 50% WPN 1.0 g/m2 was used as the molluscicide control, and the fresh water as the blank control in the field for 1 d, 3 d and 7 d, then the mortality rates of O. hupensis snails were recorded at 1 d, 3 d and 7 d after the spraying and powder-spraying. RESULTS: The snail mortality rates of LDS using the immersion test for 72 h were more than 95% in the field of eight provinces (0.1 g/m in Sichuan, Jiangxi, Jiangsu and Zhejiang provinces, 0.2 g/m3 in Yunnan, Hunan and Hubei provinces, and 0.4 g/min Anhui Province); the snail mortality rates of LDS using the spraying test for 7 d were more than 85% (0.2 g/m2 in Hunan, Hubei, Jiangxi and Zhejiang provinces, 0.4 g/m2 in Sichuan and Anhui provinces, 0.6 g/m2 in Yunnan and Jiangsu provinces). The snail mortality rates of LDS the powder-spraying test for 7 d were more than 85% (0.6 g/m2 in Yunnan, Sichuan, Hubei, Jiangxi, Anhui, Jiangsu and Zhejiang provinces). According to the standards of "Efficacy test methods and evaluation of molluscicide for pesticide registration (NY/T 1617-2008)", LDS is a qualified molluscicide. CONCLUSIONS: LDS has good molluscicidal effects through the immersion, spraying and powder-spraying test in the fields. It is suitable for a variety of environments to control O. hupensis snails of schistosomiasis endemic areas in China. The recommended dosages of LDS are 0.1-0.2 g/m3 by the immersion method, 0.2-0.4 g/m2 by the spraying method, and 0.4-0.6 g/m2 by the powder-spraying method in the fields.

Evaluating the effect of a novel molluscicide in the endemic schistosomiasis japonica area of China.

Oncomelania hupensis is the sole intermediate host snail of Schistosoma japonicum in China. Snail control by molluscicide remains one of the most effective measures of schistosomiasis japonica control. A 50% wettable powder of niclosamide ethanolamine salt (WPN) is widely used for snail control in China. However, WPN is costly and toxic to fish. A novel molluscicide named LDS, the salt of quinoid-2', 5-dichloro-4'-nitrosalicylanilide from niclosamide, has been developed. To evaluate the effects of large-scale field application of LDS on field snail control, tests were conducted in 15 counties of Hubei Province, China. Active adult snails, were immersed in 0.2, 0.4, and 0.6 g/m3 of 10% LDS, 1.0 g/m3 of 50% WPN was used as the molluscicide control, and then the mortality rates of snails were investigated after 1, 2, and 3 days. In addition, four active concentrations of 10% LDS (0.4, 0.6, 0.8 and 1.0 g/m2) were applied by spraying and powdering in the field. 1.0 g/m2 of 50% WPN was used as the molluscicide control, and then the mortality rates of snails were observed after 1, 3, and 7 days. The results indicated that 0.4 g/m3 LDS applied by the immersion or 0.6 g/m2 LDS applied by spraying and powdering achieved the same molluscicidal effect as that of WPN, regardless of exposure time. By using different methods, the snail mortality rates in the molluscicide groups were related to exposure time and concentration, respectively. LDS costs less than WPN; thus, LDS is suitable and applicable for use as a molluscicide in schistosomiasis japonica epidemic areas.