Sleep initiation patterns and sleep quality among toddlers in the southeast of China: initial study results.

BACKGROUND: A large number of psychological consequences including sleep health emerged during the 2019 Coronavirus disease (COVID-19) pandemic. Sleep patterns in toddlers are vulnerable to negative environmental exposures, however, very few studies on this topic have been published so far. OBJECTIVES: In this paper, we aimed to investigate the sleep patterns and associated factors in toddlers from China confined at home in the context of COVID-19 pandemic. METHODS: From April to November 2021, a convenience sample of 493 parents of young children aged (12-35 months) were surveyed from Fuzhou, Sanming, Quanzhou, Nanping, and Longyan cities in the Fujian Province, China. A cross-sectional survey was conducted via Electronic questionnaires to collect parent and child social-demographic characteristics. The Brief Infant Sleep Questionnaire (BISQ) was used to collect data on sleep practices, sleep duration and patterns, as well as the number of nocturnal awaking . RESULTS: The mean age of toddlers was 2.11 years old, and 52.54% (259/493) were males. Among the 493 toddlers' sleep patterns, 331(67.1%) initiated sleep accompanied by parents, 67(13.6%) slept independently, 59 (12.0%) were breast fed/bottle fed to initiate sleep, 27 (5.5%) were held and 9 (1.8%) rocked. The clear longitudinal association between the duration of night-time sleep, the frequency of nighttime awakenings, and various sleep patterns remains clear (p < 0.05). Multiple linear regression analysis indicated that sleep initiation with bottle-feeding/breast-feeding and rocked significantly increased the frequency of nighttime awakenings and reduced the duration of nighttime sleep (p < 0.05), as held was dramatically only for increasing the number of nighttime awakenings (p < 0.05). Multi-variate logistic regression analysis demonstrated that toddlers with severe sleep difficulties had a higher probability of being rocked to initiate sleep (p < 0.05). Conversely, young children with minor sleep problems were more apt to be in bed alone to initiate sleep (p < 0.05). CONCLUSIONS: During the COVID-19 pandemic, most infants and toddlers initiated sleep accompanied by parents and tend to have electronic media exposure before bedtime. Increased waking at night may be associated with sleep initiation with breast-feeding/bottle-feeding. Therefore, pediatric practitioners in primary community hospitals should pay attention to the education and promotion of sleep hygiene and parenting knowledge of young children to avoid the formation of poor sleep hygiene habits.

Irisin alleviates hyperoxia-induced bronchopulmonary dysplasia through activation of Nrf2/HO-1 pathway.

BACKGROUND: Bronchopulmonary dysplasia (BPD) is a common pulmonary injury among premature infants, which is often caused by hyperoxia exposure. Irisin is a novel hormone-like myokine derived mainly from skeletal muscles as well as adipose tissues. Many studies have indicated that Irisin exert a variety of properties against hyperoxia-induced inflammation and oxidative stress (OS). We aimed to evaluate the effects of irisin on hyperoxia-induced lung injury explore the underlying mechanisms. METHODS: BPD model was established after exposing newborn mouse to 85% oxygen. BPD mouse received continuous intraperitoneal injection of irisin at a dose of 25 µg/kg/day. Lung tissues were collected for histological examination at 7 and 14 days after birth. The alveolarization and alveolar vascularization of each animal was assessed. Levels of oxidative stress indicators, and the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in lung tissues were detected at 14 days after birth. RESULTS: Hyperoxia exposure induced a markedly alveolar simplification and a disrupted alveolar angiogenesis, which was ameliorated by irisin treatment. The hyperoxia-induced increase in these oxidative stress indicators was significantly reversed by irisin treatment. The Nrf2/HO-1 pathway is inducted in the hyperoxia-induced BPD mouse model, which is further activated by irisin treatment. CONCLUSION: Our results demonstrated the beneficial effects of irisin in reducing the OS, enhancing alveolarization, and promoting vascular development through activation of Nrf2/HO-1 axis in a hyperoxia-induced experimental model of BPD.

[Two cases of MEGDEL syndrome due to variants of SERAC1 gene and a literature review].

OBJECTIVE: To explore the clinical phenotype and genetic features of two children with MEGDEL syndrome due to variants of the SERAC1 gene. METHODS: Two children who had presented at the Fujian Medical University Union Hospital respectively on July 14, 2020 and July 28, 2018 were selected as the study subjects. Clinical features and results of genetic testing were retrospectively analyzed. RESULTS: Both children had featured developmental delay, dystonia and sensorineural deafness, along with increased urine 3-methylglutaric acid levels. Magnetic resonance imaging revealed changes similar to Leigh-like syndrome. Gene sequencing revealed that both children have harbored pathogenic compound heterozygous variants of the SERAC1 gene, including c.1159C>T and c.442C>T in child 1, and c.1168C>T and exons 4~9 deletion in child 2. CONCLUSION: Children with MEGDEL syndrome due to SERAC1 gene variants have variable clinical genotypes. Delineation of its clinical characteristics and typical imaging changes can facilitate early diagnosis and treatment. Discovery of the novel variants has also enriched the spectrum of SERAC1 gene variants.

Real-Time Vehicle Detection from UAV Aerial Images Based on Improved YOLOv5.

Aerial vehicle detection has significant applications in aerial surveillance and traffic control. The pictures captured by the UAV are characterized by many tiny objects and vehicles obscuring each other, significantly increasing the detection challenge. In the research of detecting vehicles in aerial images, there is a widespread problem of missed and false detections. Therefore, we customize a model based on YOLOv5 to be more suitable for detecting vehicles in aerial images. Firstly, we add one additional prediction head to detect smaller-scale objects. Furthermore, to keep the original features involved in the training process of the model, we introduce a Bidirectional Feature Pyramid Network (BiFPN) to fuse the feature information from various scales. Lastly, Soft-NMS (soft non-maximum suppression) is employed as a prediction frame filtering method, alleviating the missed detection due to the close alignment of vehicles. The experimental findings on the self-made dataset in this research indicate that compared with YOLOv5s, the mAP@0.5 and mAP@0.5:0.95 of YOLOv5-VTO increase by 3.7% and 4.7%, respectively, and the two indexes of accuracy and recall are also improved.

Number of positive lymph nodes combined with the logarithmic ratio of positive lymph nodes predicts long-term survival for patients with node-positive parotid gland carcinoma after surgery: a SEER population-based study.

PURPOSE: To evaluate the prognostic value of the number of positive lymph nodes (NPLN), the ratio of positive lymph nodes (pLNR), and the logarithmic ratio of positive lymph nodes (LODDS) in patients with parotid gland carcinoma. On this basis, establishing and validating an optimal nomogram. METHODS: A total of 895 patients with T1-4N1-3M0 parotid gland carcinoma were included in our study from the Surveillance, Epidemiology, and End Results (SEER) database. Patients' data were randomly assigned to the training cohort and the validation cohort by a ratio of 7:3. Univariate and multivariate COX regression analysis were used to explore the relationship between the study factors and the prognosis of parotid gland carcinoma, including overall survival (OS) and cause-specific survival (CSS). The Akaike Information Criterion (AIC) was used to evaluate model fit. Harrell's concordance index (C-index), integrated discrimination improvement (IDI), and net reclassification index (NRI) were used to evaluate the predictive ability of these models. The decision curve analysis was used to evaluate the clinical benefit of the nomograms compared with the TNM stage. RESULTS: NPLN, pLNR, and LODDS are independent risk factors for the prognostic of PGC. According to the AIC, C index, IDI, and NRI, the models combined with NPLN and LODDS were the best. The decision curves suggested that our nomograms had good predictive abilities for the prognosis of parotid gland carcinoma. CONCLUSION: The two nomograms which contained NPLN and LODDS had the potential to predict OS and CSS in patients with parotid gland carcinoma.

Prognostic Value of Controlling Nutritional Status Score in Advanced Hypopharyngeal Cancer.

OBJECTIVE: The purpose of this study was to investigate the prognostic significance of the preoperative controlling nutritional status (CONUT) score in patients with resectable advanced hypopharyngeal cancer. METHODS: This retrospective study included 113 advanced hypopharyngeal cancer patients who underwent curative resection in our hospital from 2013 to 2017. The association between the CONUT score and clinicopathological variables was evaluated. The association between CONUT score and survival was analyzed using Kaplan-Meier survival curves and Cox regression. The efficacy of the CONUT score and other immune-nutritional markers to predict prognosis was compared using a time-dependent receiver operating characteristic (ROC). RESULTS: Patients were divided into the high-CONUT score group (≥3) and the low-CONUT score group (≤2) according to ROC analysis. The CONUT score was associated with body mass index (p = 0.047), monocyte (p = 0.021), pharyngocutaneous fistula (p = 0.045), flap repairment (p = 0.034), tumor (T) classification (p = 0.034), node (N) classification (p = 0.036), subsite of tumor (p = 0.035), and negative pathologic factors (p < 0.001). Tumor, node, metastasis (TNM) stage, negative pathologic factors, adjuvant radiotherapy, postoperative chemoradiotherapy, and CONUT score were independent prognostic factors for survival. Patients with a higher CONUT score had worse overall survival (OS) (hazard ratio: 2.76, 95% confidence interval [CI]: 1.44-5.29, p = 0.002) and disease-free survival (hazard ratio: 2.51, 95% CI: 1.28-4.91, p = 0.007). The area under the curve of the CONUT score (0.799) to predict 5-year OS was greater than those of Preoperative Nutritional Index (0.769), platelet-to-lymphocyte ratio (0.643), neutrophil-to-lymphocyte ratio (0.565), and lymphocyte-to-monocyte ratio (0.577). CONCLUSION: The CONUT score is a prognostic marker for patients with resectable advanced hypopharyngeal cancer. LEVEL OF EVIDENCE: 3 Laryngoscope, 133:2613-2620, 2023.

Development of a novel electroporation method for the oyster parasite Perkinsus marinus.

Gene manipulation techniques are fundamental to molecular biology and are continuously being improved. However, gene transfection methods are not established for many unicellular eukaryotes (protists), thereby hindering molecular biological investigations. The oyster parasite Perkinisus marinus is one of the few protists with established gene transfection and drug selection. Nevertheless, the present protocols are tedious, requiring a specific electroporator and pulse conditions which limits the accessibility of this technique across different research groups. Here, we present alternative buffer and electroporation conditions that make the protocol less restrictive. We revealed the pulse condition that enables the introduction of plasmids into P. marinus cell using Ingenio electroporation buffer and NEPA21 electroporator. We found that number of cells and plasmid concentration were critical parameters for the electroporation system. We also constructed a simpler expression plasmid that is removed needless regions for gene expression in the parasite. Our findings resolved the equipment restriction in electroporation of P. marinus and would be a good reference for electroporation in other protists, in particular other Perkinsozoa parasites and core dinoflagellates.

Validation of the sleep disturbance scale for children (SDSC) in infants and toddlers from mainland China.

The sleep disturbance scale for children (SDSC) has been validated in the population of Chinese children (aged 5-16 years) and has good psychometric properties. However, valid assessment tools for sleep problems in the population of infants and young children from mainland China are relatively scarce, and rates of screening and management for pediatric sleep problems are low. Objective: This study is to evaluate the reliability and validity of SDSC among infants (aged 6-36 months), and to provide a reference for expanding the application of the SDSC for Chinese infants (SDSC-I). Materials and methods: From April to November 2021, parents of infants from Fuzhou, Quanzhou, Longyan, Sanming, and Nanping cities in Fujian Province, China completed the SDSC-I. Several factor analyses were performed to evaluate the reliability and validity of the scale. Results: Of note, 432 out of 469 samples were valid. After item selections and exploratory factor analyses, the SDSC-I concluded six dimensions (disorders of initiating sleep, disorders of maintaining sleep, sleep hyperhidrosis, sleep breathing disorders, parasomnias, and non-restorative sleep and excessive somnolence) with 23 items. The Cronbach's α coefficient of the scale was 0.863, and those for the six dimensions were within 0.576-0.835. The values of parameters for content validity of the scale were: IR = 0.87, I-CVI > 0.78, Kappa value > 0.74, S-CVI/UA = 0.87, S-CVI/Ave = 0.98. Principal component analysis revealed that the Kaiser-Meyer-Olkin (KMO) value was 0.84, and the factor loading of items ranged from 0.328 to 0.849, with six factors of eigenvalue more than one, which could explain 58.274% of the total variance. The confirmatory factor analysis results showed that χ2/DF was 3.556, root-mean-square error of approximation (RMSEA) was 0.077, comparative fit index (CFI) was 0.809, and standardized RMR (SRMR) was 0.070. Conclusion: Our study provides evidence that the SDSC-I is reliable and valid, and it is effective for the screening and management of sleep disturbances among infants (aged 6-36 months). Compared with other questionnaires such as the Brief Infant Sleep Questionnaire (BISQ), it is worthy of popularization and application in pediatric primary care.

Ultrasound combined with glial cell line-derived neurotrophic factor-loaded microbubbles for the targeted treatment of drug addiction.

Drug addiction is a serious problem globally, recently exacerbated by the COVID-19 pandemic. Glial cell-derived neurotrophic factor (GDNF) is considered a potentially effective strategy for the treatment of addiction. Previous animal experiments have proven that GDNF has a good therapeutic effect on drug addiction, but its clinical application is limited due to its poor blood-brain barrier (BBB) permeability. Low-frequency focused ultrasound, combined with microbubbles, is a non-invasive and reversible technique for locally-targeted BBB opening. In the present study, magnetic resonance imaging-guided low-frequency focused ultrasound, combined with GDNF microbubbles, was used to target BBB opening in the ventral tegmental area (VTA) region. The effects of GDNF on morphine-induced conditioned place preference (CPP) and acute withdrawal symptoms in rats after a partially opened BBB were evaluated by behavioral observation. Western blot was used to detect changes in tyrosine hydroxylase (TH) expression levels in the VTA region after different treatments, and high performance liquid chromatography was used to detect the changes in monoamine neurotransmitter content. The results showed that ultrasound combined with GDNF microbubbles targeted and opened the BBB in the VTA region, and significantly increased GDNF content, destroyed morphine-induced CPP, and reduced the withdrawal symptoms of morphine addiction in rats. Furthermore, the up-regulation of TH expression and the increase of norepinephrine and dopamine content induced by morphine were significantly reversed, and the increase of 5-hydroxytryptamine content was partially reversed. Therefore, ultrasound combined with GDNF microbubbles to target and open the BBB can effectively increase the content of central GDNF, thus playing a therapeutic role in morphine addiction. Our study provides a new approach to locally open the BBB and target delivery of neurotrophic factors, such as GDNF, to treat brain diseases like addiction.

The effect of droplet size on syntrophic dynamics in droplet-enabled microbial co-cultivation.

Co-cultivation in microfluidic droplets has emerged as a versatile tool for the study of natural and synthetic microbial communities. In particular, the identification and characterization of syntrophic interactions in these communities is attracting increasing interest due to their critical importance for the functioning of environmental and host-associated communities as well as new biotechnological applications. However, one critical parameter in droplet-enabled co-cultivation that has evaded appropriate evaluation is the droplet size. Given the same number of initial cells, a larger droplet size can increase the length scale secreted metabolites must diffuse as well as dilute the initial concentration of cells and exchanged metabolites, impacting the community dynamics. To evaluate the effect of droplet size on a spectrum of syntrophic interactions, we cultivated a synthetic model system consisting of two E. coli auxotrophs, whose interactions could be modulated through supplementation of related amino acids in the medium. Our results demonstrate that the droplet size impacts substantially numerous aspects of the growth of a cross-feeding bi-culture, particularly the growth capacity, maximum specific growth rate, and lag time, depending on the degree of the interaction. This work heavily suggests that one droplet size does not fit all types of interactions; this parameter should be carefully evaluated and chosen in experimental studies that aim to utilize droplet-enabled co-cultivation to characterize or elucidate microbial interactions.

Imiquimod induced vitiligo-like lesions-A consequence of modified melanocyte function.

INTRODUCTION: Imiquimod plays an important role in the management of condyloma and premalignant lesions. Successively, an increase of hypopigmented lesions following imiquimod application has been reported. However, the mechanisms of imiquimod on melanocytes remain unclear. This study was designed to assess the effect of Imiquimod on the functions of melanocytes in vitro. METHODS: Primary cultured melanocytes were isolated from normal control skin tissue. After incubation with imiquimod for 48 h in vitro, cell viability was analyzed by cell counting kit-8 assay. Apoptosis was detected using the Annexin V-fluorescein-5-isothiocyanate flow cytometry assay. Melanin content and tyrosinase activity in melanocytes were measured by colorimetric method and the modified dopachrome method. The production of inflammatory cytokine interleukin 8 (IL-8), IL-6, and soluble ICAM-1 (soluble Intercellular Adhesion Molecule-1[sICAM-1]) in melanocytes were measured by enzyme-linked immunosorbent assay (ELISA). Toll-like receptor 7 (TLR7), toll-like receptor 9 (TLR9) protein, and autophagy-related proteins microtubule-associated protein 1A/1B-light chain 3 (LC3-II), p62, mechanistic target of rapamycin (mTOR), and Atg5 were assessed using western blot analysis. RESULTS: Imiquimod significantly inhibited the activity of tyrosinase activity and decreased melanin content in melanocytes and significantly increased apoptosis and IL-6, IL-8, and sICAM-1 production in melanocytes. Moreover, the expression of TLR7 and TLR9 proteins were significantly increased, and the expression of mTOR, p62 protein were markedly decreased, but the expression of LC3II/I and Atg5 protein were significantly increased in melanocytes after incubating with imiquimod. CONCLUSIONS: This study shows that imiquimod directly inhibits melanogenesis and increases melanocyte apoptosis rates. These effects combined with the upregulation of TLR7 and TLR9 together with increased autophagy activity and inflammatory cytokines production, might be the main reasons leading to hypopigmented lesions after imiquimod application.

Disturbed relationship between glucocorticoid receptor and 5-HT1AR/5-HT2AR in ADHD rats: A correlation study.

Objective: This work is to investigate the alterations of the central 5-hydroxytryptamine (5-HT) system in spontaneously hypertensive rats (SHR) and the correlation with the behaviors of SHR, and to explore the effects of glucocorticoid intervention on the central 5-HT system and SHR behaviors. Materials and methods: Three weeks old SHR were chosen as the attention-deficit hyperactivity disorder (ADHD) model and treated with glucocorticoid receptor (GR) agonist or inhibitor, whereas Wista Kyoto rats (WKY) were chosen as the normal control group. Open-field test and Làt maze test were used to evaluate the spontaneous activities and non-selective attention. The levels of 5-HT in the extracellular fluid specimens of the prefrontal cortex of rats were analyzed by high-performance liquid chromatography. The expressions of GR, 5-HT1A receptor (5-HT1AR), and 5-HT2A receptor (5-HT2AR) in the prefrontal cortex were analyzed through immunohistochemistry. Results: Our study demonstrated that the 5-HT level was lower in the prefrontal cortex of SHR compared to that of WKY. The Open-field test and Làt maze test showed that GR agonist (dexamethasone, DEX) intervention ameliorated attention deficit and hyperactive behavior, whereas GR inhibitor (RU486) aggravated the disorders. With DEX, the expression levels of 5-HT and 5-HT2AR in the prefrontal cortex of SHR were significantly higher than those in the control group, whereas the expression level of 5-HT1AR was lower. However, the expression levels of 5-HT and 5-HT2AR were significantly decreased after the intervention with RU486, while the expression level of 5-HT1AR increased. Results showed that glucocorticoid was negatively correlated with 5-HT1AR and positively correlated with 5-HT2AR. Conclusion: In the prefrontal cortex of ADHD rats, the down-regulation of 5-HT and 5-HT2AR expressions and the up-regulation of 5-HT1AR, compared with WYK rats, suggested a dysfunctional central 5-HT system in ADHD rats. The GR agonist can upregulate the expression of 5-HT and 5-HT2AR and downregulate the expression of 5-HT1AR in the prefrontal cortex of SHR as well as reduce the hyperactivity and attention deficit behavior in SHR, while the opposite was true for the GR inhibitor. It is suggested that the dysfunction of the 5-HT system in ADHD rats is closely related to glucocorticoid receptor activity.

The Difference in Expression of Autophagy-Related Proteins in Lesional and Perilesional Skin in Adult Patients with Active and Stable Generalized Vitiligo-A Cross-Sectional Pilot Study.

BACKGROUND: Autophagy plays an important role in maintaining intracellular homeostasis and is essential for cell survival and cell death. Dysfunction of autophagy has been described in many autoimmune diseases but data on vitiligo are scarce. AIMS: The aim of this pilot study was to investigate the expression of autophagy-related proteins in patients with vitiligo. METHODS: Western blotting was used to analyze the expression of microtubule-associated protein light chain 3 (LC3II/I), autophagy-related gene 5 (Agt5), mammalian target of rapamycin (mTOR) and p62 in lesional and perilesional vitiligo skin from seven patients with active generalized vitiligo and nine patients with stable generalized vitiligo compared to control skin from six healthy subjects. RESULTS: Our data showed increased expression of the autophagy marker LC3II/I and decreased p62 protein expression in lesional skin of active and stable vitiligo compared to control skin (P < 0.01). No significant difference in the expression of LC3II/I and p62 was found in perilesional skin of active vitiligo patients (P > 0.05) compared to control skin. Expression of LC3II/I in stable vitiligo lesional skin was higher and p62 expression was lower compared to active vitiligo lesional skin (P < 0.01). Decreased p62 expression was shown in perilesional skin of stable vitiligo patients (P < 0.05). Agt5 protein in lesional and perilesional skin of both active and stable vitiligo patients were increased (P < 0.01 and P< 0.05) compared to control skin. The expression of mTOR protein in lesional and perilesional skin of active and stable vitiligo patients was significantly lower than in control skin (P < 0.01). CONCLUSIONS: The present study indicates increased autophagy in lesional skin in vitiligo patients. Stable vitiligo lesional skin showed increased autophagy compared to active vitiligo lesional skin. Missing activation of autophagy in active vitiligo perilesional skin suggests disturbed autophagy to be associated with vitiligo.

Lectin receptor-like kinase LecRK-VIII.2 is a missing link in MAPK signaling-mediated yield control.

The energy allocation for vegetative and reproductive growth is regulated by developmental signals and environmental cues, which subsequently affects seed output. However, the molecular mechanism underlying how plants coordinate yield-related traits to control yield in changing source-sink relationships remains largely unknown. Here, we discovered the lectin receptor-like kinase LecRK-VIII.2 as a specific receptor-like kinase that coordinates silique number, seed size, and seed number to determine seed yield in Arabidopsis (Arabidopsis thaliana). The lecrk-VIII.2 mutants develop smaller seeds, but more siliques and seeds, leading to increased yield. In contrast, the plants overexpressing LecRK-VIII.2 form bigger seeds, but less siliques and seeds, which results in similar yield to that of wild-type plants. Interestingly, LecRK-VIII.2 promotes the growth of the rosette, root, and stem by coordinating the source-sink relationship. Additionally, LecRK-VIII.2 positively regulates cell expansion and proliferation in the seed coat, and maternally controls seed size. The genetic and biochemical analyses demonstrated that LecRK-VIII.2 acts upstream of the mitogen-activated protein kinase (MAPK) gene MPK6 to regulate silique number, seed size, and seed number. Collectively, these findings uncover LecRK-VIII.2 as an upstream component of the MAPK signaling pathway to control yield-related traits and suggest its potential for crop improvement aimed at developing plants with stable yield, a robust root system, and improved lodging resistance.

The prevalence of sleep problems among children in mainland China: a meta-analysis and systemic-analysis.

BACKGROUND: We conducted a meta-analysis and systematic review to identify a reliable estimate of sleep problems prevalence among children in mainland China and to describe its epidemiological characteristics. METHODS: Relevant studies were searched thoroughly via electronic databases included China National Knowledge Infrastructure, Wanfang, Weipu, PubMed, Embase and Medline databases from inception until December 2020. Prevalence estimates were calculated by random-effects models. The sources of heterogeneity were explored using subgroup analyses and Meta-regression analysis, and publication bias was estimated by funnel plots and Egger's Test. RESULTS: Overall, 66 studies were included in this meta-analysis, which revealed that the pooled prevalence of sleep problems was 37.6% (95%CI: 34.3-40.9%) with high heterogeneity (I2 = 99.6%,P < 0.001). The incidence of snoring was 7.7%, choking or gasping was 0.9%, apnea was 1.5%, restless sleep was 11.3%, mouth breathing was 4.7%, hyperhidrosis was 17.2%, leg movements was 2.7%, bruxism was 9.6%, sleep talking was 0.7%, sleep-walking was 0.8%, nightmare was 5.1%, enuresis was 3.4%, night awakening was 6.7% and trouble falling asleep was 11.1%. The prevalence rate of sleep problems among males was higher than females (OR:1.01,95%CI:1.05-1.13). In all age groups, the prevalence rates of sleep problems increased with age, including infancy or early childhood group (33.3%), pre-school group (38.9%), school-age group (43.7%). The prevalence rate in South China (30.4%, 95%CI: 23.9-36.8%) was the lowest, and the highest prevalence rate was in West China (47.4%,95%CI:35.9-58.9%), which than any other region in China. The point estimate for sleep disorders prevalence obtained using the CSHQ criterion was higher than other criteria. Meta-regression indicated that age group could influence prevalence estimation (P = 0.011). CONCLUSIONS: Over the past two decades, the prevalence rate of sleep problems among children in mainland China has increased, significantly affecting two-fifth of the school-age children. The incidence of hyperhidrosis, restless sleep and trouble falling asleep were significantly higher than other sleep prombles. The prevalence rate of sleep problems in west China was significantly higher than in any other area. There is still a lack of guidelines on children's sleep problems in mainland China, so future research should pay special attention to the sleep problems of school-age children and children in economically backward areas.

Individual Microcystis colonies harbour distinct bacterial communities that differ by Microcystis oligotype and with time.

Interactions between bacteria and phytoplankton in the phycosphere have impacts at the scale of whole ecosystems, including the development of harmful algal blooms. The cyanobacterium Microcystis causes toxic blooms that threaten freshwater ecosystems and human health globally. Microcystis grows in colonies that harbour dense assemblages of other bacteria, yet the taxonomic composition of these phycosphere communities and the nature of their interactions with Microcystis are not well characterized. To identify the taxa and compositional variance within Microcystis phycosphere communities, we performed 16S rRNA V4 region amplicon sequencing on individual Microcystis colonies collected biweekly via high-throughput droplet encapsulation during a western Lake Erie cyanobacterial bloom. The Microcystis phycosphere communities were distinct from microbial communities in whole water and bulk phytoplankton seston in western Lake Erie but lacked 'core' taxa found across all colonies. However, dissimilarity in phycosphere community composition correlated with sampling date and the Microcystis 16S rRNA oligotype. Several taxa in the phycosphere were specific to and conserved with Microcystis of a single oligotype or sampling date. Together, this suggests that physiological differences between Microcystis strains, temporal changes in strain phenotypes, and the composition of seeding communities may impact community composition of the Microcystis phycosphere.

Optimized gene expression from bacterial chromosome by high-throughput integration and screening.

Chromosomal integration of recombinant genes is desirable compared with expression from plasmids due to increased stability, reduced cell-to-cell variability, and elimination of the need for antibiotics for plasmid maintenance. Here, we present a new approach for tuning pathway gene expression levels via random integration and high-throughput screening. We demonstrate multiplexed gene integration and expression-level optimization for isobutanol production in Escherichia coli The integrated strains could, with far lower expression levels than plasmid-based expression, produce high titers (10.0 ± 0.9 g/liter isobutanol in 48 hours) and yields (69% of the theoretical maximum). Close examination of pathway expression in the top-performing, as well as other isolates, reveals the complexity of cellular metabolism and regulation, underscoring the need for precise optimization while integrating pathway genes into the chromosome. We expect this method for pathway integration and optimization can be readily extended to a wide range of pathways and chassis to create robust and efficient production strains.

The specific DNA barcodes based on chloroplast genes for species identification of Orchidaceae plants.

DNA barcoding is currently an effective and widely used tool that enables rapid and accurate identification of plant species. The Orchidaceae is the second largest family of flowering plants, with more than 700 genera and 20,000 species distributed nearly worldwide. The accurate identification of Orchids not only contributes to the safe utilization of these plants, but also it is essential to the protection and utilization of germplasm resources. In this study, the DNA barcoding of 4 chloroplast genes (matK, rbcL, ndhF and ycf1) were used to provide theoretical basis for species identification, germplasm conservation and innovative utilization of orchids. By comparing the nucleotide replacement saturation of the single or combined sequences among the 4 genes, we found that these sequences reached a saturation state and were suitable for phylogenetic relationship analysis. The phylogenetic analyses based on genetic distance indicated that ndhF and ycf1 sequences were competent to identification at genus and species level of orchids in a single gene. In the combined sequences, matK + ycf1 and ndhF + ycf1 were qualified for identification at the genera and species levels, suggesting the potential roles of ndhF, ycf1, matK + ycf1 and ndhF + ycf1 as candidate barcodes for orchids. Based on the SNP sites, candidate genes were used to obtain the specific barcode of orchid plant species and generated the corresponding DNA QR code ID card that could be immediately recognized by electronic devices. This study provides innovative research methods for efficient species identification of orchids. The standardized and accurate barcode information of Orchids is provided for researchers. It lays the foundation for the conservation, evaluation, innovative utilization and protection of Orchidaceae germplasm resources.