Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs.

Conversely to canonical splicing, back-splicing connects the upstream 3' splice site (SS) with a downstream 5'SS and generates exonic circular RNAs (circRNAs) that are widely identified and have regulatory functions in eukaryotic gene expression. However, sex-specific back-splicing in Drosophila has not been investigated and its regulation remains unclear. Here, we performed multiple RNA analyses of a variety sex-specific Drosophila samples and identified over ten thousand circular RNAs, in which hundreds are sex-differentially and -specifically back-spliced. Intriguingly, we found that expression of SXL, an RNA-binding protein encoded by Sex-lethal (Sxl), the master Drosophila sex-determination gene that is only spliced into functional proteins in females, promoted back-splicing of many female-differential circRNAs in the male S2 cells, whereas expression of a SXL mutant (SXLRRM) did not promote those events. Using a monoclonal antibody, we further obtained the transcriptome-wide RNA-binding sites of SXL through PAR-CLIP. After splicing assay of mini-genes with mutations in the SXL-binding sites, we revealed that SXL-binding on flanking exons and introns of pre-mRNAs facilitates back-splicing, whereas SXL-binding on the circRNA exons inhibits back-splicing. This study provides strong evidence that SXL has a regulatory role in back-splicing to generate sex-specific and -differential circRNAs, as well as in the initiation of sex-determination cascade through canonical forward-splicing.

[Hirsutine induces apoptosis of human breast cancer MDA-MB-231 cells through mitochondrial pathway].

The aim of this study was to investigate the effects of hirsutine on apoptosis of breast cancer cells and its possible mechanism. The MCF-10A, MCF-7 and MDA-MB-231 cells were treated with hirsutine at different concentrations for 48 h or incubated with 160 µmol/L hirsutine for 24, 48, and 72 h. The MCF-10A cell line is a non-tumorigenic epithelial cell line, and the MCF-7 and MDA-MB-231 are human breast adenocarcinoma cell lines. CCK-8 assay was employed to detect the cell viability. Flow cytometry was used to assay the apoptosis and mitochondrial membrane potential (MMP). The protein expressions of Bcl-2, Bax, cleaved-caspase 9, cleaved-caspase 3 and cytochrome C (Cyt C) in the MDA-MB-231 cells were detected by Western blotting. The results showed that hirsutine remarkably reduced the viability of MCF-7 and MDA-MB-231 cells in a time- and dose-dependent manner (P < 0.05) with IC50 values of 447.79 and 179.06 µmol/L, respectively. In the MDA-MB-231 cells, hirsutine induced apoptosis and depolarization of MMP (P < 0.05), released Cyt C from mitochondria (P < 0.05), and activated caspase 9 and caspase 3 (P < 0.05). However, these effects induced by hirsutine were all inhibited by cyclosporin A (CsA) (P < 0.05), a specific inhibitor of mitochondrial permeability transition pore (MPTP). In addition, hirsutine down-regulated the protein level of Bcl-2 and up-regulated the protein level of Bax (P < 0.05). These results suggest that hirsutine may induce apoptosis of human breast cancer MDA-MB-231 cells through decreasing the ratio of Bcl-2 to Bax, opening MPTP, releasing Cyt C from mitochondria, and activating caspase 9 and caspase 3.

Reduced glutathione and glutathione disulfide in the blood of glucose-6-phosphate dehydrogenase-deficient newborns.

BACKGROUND: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is commonly detected during mass screening for neonatal disease. We developed a method to measure reduced glutathione (GSH) and glutathione disulfide (GSSG) using tandem mass spectrometry (MS/MS) for detecting G6PD deficiency. METHODS: The concentration of GSH and the GSH/GSSG ratio in newborn dry-blood-spot (DBS) screening and in blood plus sodium citrate for test confirmation were examined by MS/MS using labeled glycine as an internal standard. RESULTS: G6PD-deficient newborns had a lower GSH content (242.9 ± 15.9 µmol/L)and GSH/GSSG ratio (14.9 ± 7.2) than neonatal controls (370.0 ± 53.2 µmol/L and 46.7 ± 19.6, respectively). Although the results showed a significance of P < 0.001 for DBS samples plus sodium citrate that were examined the first day after preparation, there were no significant differences in the mean GSH concentration and GSH/GSSG ratio between the G6PD deficiency-positive and negative groups when examined three days after sample preparation. CONCLUSION: The concentration of GSH and the ratio of GSH/GSSG in blood measured using MS/MS on the first day of sample preparation are consistent with G6PD activity and are helpful for diagnosing G6PD deficiency.

Minor Spliceosomal 65K/RNPC3 Interacts with ANKRD11 and Mediates HDAC3-Regulated Histone Deacetylation and Transcription.

RNA splicing is crucial in the multilayer regulatory networks for gene expression, making functional interactions with DNA- and other RNA-processing machineries in the nucleus. However, these established couplings are all major spliceosome-related; whether the minor spliceosome is involved remains unclear. Here, through affinity purification using Drosophila lysates, an interaction is identified between the minor spliceosomal 65K/RNPC3 and ANKRD11, a cofactor of histone deacetylase 3 (HDAC3). Using a CRISPR/Cas9 system, Deletion strains are constructed and found that both Dm65KΔ/Δ and Dmankrd11Δ/Δ mutants have reduced histone deacetylation at Lys9 of histone H3 (H3K9) and Lys5 of histone H4 (H4K5) in their heads, exhibiting various neural-related defects. The 65K-ANKRD11 interaction is also conserved in human cells, and the HsANKRD11 middle-uncharacterized domain mediates Hs65K association with HDAC3. Cleavage under targets and tagmentation (CUT&Tag) assays revealed that HsANKRD11 is a bridging factor, which facilitates the synergistic common chromatin-binding of HDAC3 and Hs65K. Knockdown (KD) of HsANKRD11 simultaneously decreased their common binding, resulting in reduced deacetylation of nearby H3K9. Ultimately, this study demonstrates that expression changes of many genes caused by HsANKRD11-KD are due to the decreased common chromatin-binding of HDAC3 and Hs65K and subsequently reduced deacetylation of H3K9, illustrating a novel and conserved coupling mechanism that links the histone deacetylation with minor spliceosome for the regulation of gene expression.

Health-related quality-of-life after traumatic brain injury: a 2-year follow-up study in Wuhan, China.

OBJECTIVE: To assess health-related quality-of-life (HRQoL) 2 years after traumatic brain injury (TBI) among a group of Chinese. METHODS: A total of 358 adult patients with moderate-to-severe TBI based on Glasgow Coma Scale score were recruited in a large trauma centre in Wuhan, China during May 2005 to April 2008. They were followed up for 2 years and the Medical Outcome Short Form 36 was used to measure HRQoL. RESULTS: After a 2-year follow-up, there were 312 (87.2%) survivors. All domains of HRQoL had the lowest scores at discharge, greatly improved over the first 6 months and showed continued improvement. Patients with TBI still had significantly lower scores in every domain than the reference group 2 years after discharge. Female patients had lower MCS scores than the males (OR = 1.8, 95% CI: 1.1-2.9). Patients older than 30 had lower scores in PCS (OR = 1.7, 95% CI: 1.1-2.6). Patients with severe TBI had lower scores in both PCS (OR = 1.9, 95% CI: 1.2-3.1) and MCS (OR = 1.6, 95% CI: 1.0-2.6) compared with those with moderate TBI. CONCLUSIONS: HRQoL of a group of Chinese patients with TBI improved during 2 years after discharge. Age, sex and severity of TBI were significantly associated with physical or mental HRQoL after discharge.

A Fully Flexible Intelligent Thermal Touch Panel Based on Intrinsically Plastic Ag2 S Semiconductor.

Wearable touch panels, a typical flexible electronic device, can recognize and feed back the information of finger touch and movement. Excellent wearable touch panels are required to accurately and quickly monitor the signals of finger movement as well as the capacity of bearing various types of deformation. High-performance thermistor materials are one of the key functional components, but to date, a long-standing bottleneck is that inorganic semiconductors are typically brittle while the electrical properties of organic semiconductors are quite low. Herein, a high-performance flexible temperature sensor is reported by using plastic Ag2 S with ultrahigh temperature coefficient of resistance of -4.7% K-1 and resolution of 0.05 K, and rapid response/recovery time of 0.11/0.11 s. Moreover, the temperature sensor shows excellent durability without performance damage or loss during force stimuli tests. In addition, a fully flexible intelligent touch panel composed of a 16 × 10 Ag2 S-film-based temperature sensor array, as well as a flexible printed circuit board and a deep-learning algorithm is designed for perceiving finger touch signals in real-time, and intelligent feedback of Chinese characters and letters on an app. These results strongly show that high-performance flexible inorganic semiconductors can be widely used in flexible electronics.

[Evaluation of risk factors for retinopathy of prematurity].

OBJECTIVE: To evaluate the incidence and risk factors of ROP (retinopathy of prematurity) through a prospective multicenter study. METHODS: Eleven children's or maternal & child hospitals participated in a collective network. All infants of birth weight < 2000 g, born in or transferred to one of the participating centers from January 1st 2005 to February 28th 2006 were recruited. Timely ophthalmologic examinations were performed. The relevant data at baseline and endpoints were collected at each unit. RESULTS: A total of 882 preterm infants fulfilled the screening criteria and 752 finished a followup. And 123 infants (16.4%) had some degree of ROP. Infants with ROP had a lower gestational age, birth weight and a longer duration of oxygen versus those without ROP [(30.82 +/- 0.20) weeks vs (32.56 +/- 0.09) weeks, (1430 +/- 25) g vs (1636 +/-10) g, (11.6 +/- 1.4) d vs (4.4 +/- 0.3) d]. Through a univariate analysis, birth weight, gestational age, asphyxia, oxygen duration > 5 days, apnea, surfactant usage, pneumonia, anemia, blood transfusion, acidosis and neonatal respiratory distress syndrome (NRDS) were associated with ROP (P < 0.05). Logistic regression analysis showed that birth weight (OR = 0.998), apnea (OR = 1.653) and blood transfusion (OR = 1.763) were independent risk factors for ROP. CONCLUSION: Asphyxia, oxygen duration > 5 days, surfactant usage, anemia, acidosis and NRDS, lower birth weight, apnea and blood transfusion may improve the risks of ROP.

In vitro physiological responses of Fusarium oxysporum f. sp. niveum to exogenously applied syringic acid.

Plant-microbe interactions are often accompanied by allelochemicals, such as syringic acid, released from the host plant. To explore the role of phenolic acids released from crop host plants in response to pathogen invasion, we examined the allelopathic effect of an artificially applied syringic acid on Fusarium oxysporum f. sp. niveum. We demonstrated that the growth and the conidial germination rate of F. oxysporum f. sp. niveum were stimulated at lower concentrations of syringic acid, though inhibited by higher dosage compared with control. The yield of fungus mycotoxin was increased from 60.9% to 561.5%. We conclude that syringic acid can be considered as a allelochemical inducer, stimulating the relative virulence factors of invading pathogens.

Polymorphisms in DNA repair gene XRCC1 and increased genetic susceptibility to glioma.

BACKGROUND: The XRCC1 gene encodes the XRCC1 protein, which complexes with three other DNA repair enzymes involved in the base-excision repair (BER) pathways. Different XRCC1 polymorphisms may increase the risk of cancers by impairing interaction with other enzymatic proteins and consequently altering DNA repair activity, and result in carcinogenesis. Our study aimed to investigate any association between three polymorphisms of the XRCC1 gene at codon 194, 280 and 399 and potential glioma risk. METHODS: We collected 127 patients with primary glioma and 249 controls who requested general health examinations from Union Hospital of Tongji Medical College hospital from March 2007 to September 2010. A total of 5 ml venous blood was drawn from each subject. The polymorphisms of XRCC1 gene at codons 194, 280 and 399 were analyzed based on duplex polymerase-chain-reactions with the confronting-two-pair primer (PCR-CTPP) method. RESULTS: The homozygous Trp/Trp and heterozygotes Arg/Trp variants of codon 194 had a 2.12 fold and 1.46 fold increased risk of glioma compared to the homozygous Arg/Arg wide genotypes. The same effect was found in codon 399, the codon 399 Gln/Gln and Arg/Gln genotypes being associated with a 2.24 fold and 1.67 fold increased risk in glioma. When comparing the codon 194 Arg/Arg and 399 Arg/Arg genotypes, the combination of codon 194 Trp allele and 399 Gln allele had a heavy increase in glioma risk (OR=2.87, 95%CI=1.56-6.73). CONCLUSION: The present study provided evidence of a potential role for XRCC1 codon 194 and 399 polymorphisms in genetic predisposition to glioma among the Chinese population. This analysis of correlation of DNA repair genes and glioma may provide a deeper insight into the genetic and environment factors for cancer risk.

[Responses of rice-wheat rotation system in south Jiangsu to organic-inorganic compound fertilizers].

In 2006-2007, a field trial was conducted to study the effects of applying three kinds of organic-inorganic compound fertilizers [rapeseed cake compost plus inorganic fertilizers (RCC), pig manure compost plus inorganic fertilizers (PMC), and Chinese medicine residues plus inorganic fertilizers (CMC)] on the crop growth and nitrogen (N) use efficiency of rice-wheat rotation system in South Jiangsu. Grain yield of wheat and rice in the different fertilization treatments was significantly higher than the control (no fertilization). In treatments RCC, PMC and CMC, the wheat yield was 13.1%, 32.2% and 39.3% lower than that of the NPK compound fertilizer (CF, 6760 kg x hm(-2)), respectively, but the rice yield (8504-9449 kg x hm(-2)) was significantly higher than that (7919 kg x hm(-2)) of CF, with an increment of 7.4%-19.3%. In wheat season, the aboveground dry mass, N accumulation, and N use efficiency in treatments RCC, PMC, and CMC were lower than those of CF, but in rice season, these parameters were significantly higher than or as the same as CF. In sum, all the test three compound fertilizers had positive effects on the rice yield and its nitrogen use efficiency in the rice-wheat rotation system, being most significant for RCC.

[Effects of organic-inorganic mixed fertilizers on rice yield and nitrogen use efficiency].

A field experiment was carried to study the effects of organic-inorganic mixed fertilizers on rice yield, nitrogen (N) use efficiency, soil N supply, and soil microbial diversity. Rapeseed cake compost (RCC), pig manure compost (PMC), and Chinese medicine residue compost (MRC) were mixed with chemical N, P and K fertilizers. All the treatments except CK received the same rate of N. The results showed that all N fertilizer application treatments had higher rice yield (7918.8-9449.2 kg x hm(-2)) than the control (6947.9 kg x hm(-2)). Compared with that of chemical fertilizers (CF) treatment (7918.8 kg x hm(-2)), the yield of the three organic-inorganic mixed fertilizers treatments ranged in 8532.0-9449.2 kg x hm(-2), and the increment was 7.7%-19.3%. Compared with treatment CF, the treatments of organic-inorganic mixed fertilizers were significantly higher in N accumulation, N transportation efficiency, N recovery rate, agronomic N use efficiency, and physiological N use efficiency. These mixed fertilizers treatments promoted rice N uptake and improved soil N supply, and thus, increased N use efficiency, compared with treatments CF and CK. Neighbor joining analysis indicated that soil bacterial communities in the five treatments could be classified into three categories, i.e., CF and CK, PMC and MRC, and RCC, implying that the application of exogenous organic materials could affect soil bacterial communities, while applying chemical fertilizers had little effect on them.

[Application of electrical impedance equipment in evaluating local lung volume during different PEEP ventilation and at different respiratory rate].

OBJECTIVE: To evaluate the applicability of electrical impedance equipment in assessing local lung volume during different PEEP ventilation and at different respiratory rate in neonatal piglets. METHODS: Electrical impedance measurements (EIM) were performed on 6 healthy newborn piglets (age 4 +/- 1 d, weight 1.66 +/- 0.31 kg) using 8 electrodes distributed to 4 quadrants of the lung (left, right, upper, lower). Tidal impedance and functional residual impedance changes during PEEP levels of 2, 4, 6 and 8 cm H2O and frequencies ranging from 0.5 to 15 Hz were investigated. RESULTS: The sum of regional tidal impedance obtained from four quadrants, significantly reflected tidal volume (VT) measured by a pneumotachograph during both frequency and PEEP changed (r2 = 0.98). A decrease of PEEP 4 to 2 cm H2O caused a significant increase in total tidal impedance (TTI) as well as in VT (P < 0.01 and P < 0.05); whereas an increase in frequency from 0.5 to 15 Hz was associated with a significant decrease in both TTI and VT (P < 0.05 and P < 0.01). Increased frequency had a balancing effect on air distribution, whereas higher PEEP did not result in more homogeneous ventilation. Minimal impedance values (FRI) (surrogate variable for FRC) showed that total FRI significantly decreased with increasing PEEP level. CONCLUSIONS: EIM demonstrated good applicability to assess changes in thoracic gas volume. It is highly suggested that this method could be considered and further studied as a non-invasive bedside method to monitor continuously regional lung ventilation of neonates under any mode of mechanical ventilation.

Antibiotic effect of exogenously applied salicylic acid on in vitro soilborne pathogen, Fusarium oxysporum f.sp.niveum.

Salicylic acid, which is biosynthesized inside plant and is often found and accumulated in soil due to plant debris decaying, is considered as a signaling substance during plant-microbe interactions. It is involved in the cycling of biogeochemistry and related to plant resistance to biotic and abiotic stress. The antibiotic effect of salicylic acid on Fusarium oxysporum f.sp.niveum (FON) was studied to investigate the relationships between the salicylic acid and the fungus in the ecological interaction of plant-microbe. Results showed that the biomass, colony diameter, number of conidium germination and conidium production of FON were decreased by 52.0%, 25.7%, 100% and 100% at concentrations of 800 mg L(-1). However, mycotoxin yield was increased by 233%, pectinase activity raised by 168.0% and cellulase activity increased by 1325% compared to control at higher concentrations. It was concluded that salicylic acid as an allelochemical greatly inhibited FON growth and conidia formation and germination, though stimulated mycotoxin production and activities of hydrolytic enzymes by FON.