Genes involved in melanin production directly impact insect pigmentation and can affect diverse physiology and behaviours. The role these genes have on sex behaviour, however, is unclear. In the present study, the crucial melanin pigment gene black was functionally characterised in an urban pest, the German cockroach, Blattella germanica. RNAi knockdown of B. germanica black (Bgblack) had no effect on survival, but did result in black pigmentation of the thoraxes, abdomens, heads, wings, legs, antennae, and cerci due to cuticular accumulation of melanin. Sex-specific variation in the pigmentation pattern was apparent, with females exhibiting darker coloration on the abdomen and thorax than males. Bgblack knockdown also resulted in wing deformation and negatively impacted the contact sex pheromone-based courtship behaviour of males. This study provides evidence for black function in multiple aspects of B. germanica biology and opens new avenues of exploration for novel pest control strategies.

Iodine is an essential nutrient that may change the occurrence of autoimmune thyroiditis (AIT). Apoptosis and DNA methylation participate in the pathogenesis and destructive mechanism of AIT. We detected the methylation and the expression of mRNA of intrinsic apoptosis-associated genes (YWHAG, ING4, BRSK2 and GJA1) to identify the potential interactions between the levels of methylation in these genes and different levels of iodine. 176 adult patients with AIT in Shandong Province, China, were included. The MethylTargetTM assay was used to verify the levels of methylation. We used PCR to detect the mRNA levels of the candidate genes. Interactions between methylation levels of the candidate genes and iodine levels were evaluated with multiplicative and addictive interaction models and GMDR. In the AIT group, YWHAG_1 and six CpG sites and BRSK2_1 and eight CpG sites were hypermethylated, whereas ING4_1 and one CpG site were hypomethylated. A negative correlation was found between methylation levels of YWHAG and mRNA expression. The combination of iodine fortification, YWHAG_1 hypermethylation and BRSK2_1 hypermethylation was significantly associated with elevated AIT risk. A four-locus model (YWHAG_1 × ING4_1 × BRSK2_1 × iodine level) was found to be the best model of the gene–environment interactions. We identified abnormal changes in the methylation status of YWHAG, ING4 and BRSK2 in patients with AIT in different iodine levels. Iodine fortification not only affected the methylation levels of YWHAG and BRSK2 but also interacted with the methylation levels of these genes and may ultimately increase the risk of AIT.

Meristems in land plants share conserved functions but develop highly variable structures. Meristems in seed-free plants, including ferns, usually contain one or a few pyramid-/wedge-shaped apical cells (ACs) as initials, which are lacking in seed plants. It remained unclear how ACs promote cell proliferation in fern gametophytes and whether any persistent AC exists to sustain fern gametophyte development continuously. Here, we uncovered previously undefined ACs maintained even at late developmental stages in fern gametophytes. Through quantitative live-imaging, we determined division patterns and growth dynamics that maintain the persistent AC in Sphenomeris chinensis, a representative fern. The AC and its immediate progenies form a conserved cell packet, driving cell proliferation and prothallus expansion. At the apical centre of gametophytes, the AC and its adjacent progenies display small dimensions resulting from active cell division instead of reduced cell expansion. These findings provide insight into diversified meristem development in land plants.

A new system for preparing 14C samples was established for a compact accelerator mass spectrometer (GXNU-AMS) at Guangxi Normal University. This sample preparation system consists of three units: a vacuum maintenance unit, a CO2 purification unit, and a CO2 reduction unit, all of which were made of quartz glass. A series of radiocarbon (14C) preparation experiments were conducted to verify the reliability of the system. The recovery rate of graphite obtained was more than 80%. The carbon content in the commercial toner and wood sample was linearly fitted to the CO2 pressure in the measurement unit of the system. The results showed a good linear relationship, indicating that the reliability of the sample preparation system. AMS measurements were conducted on a batch of standard, wood, and dead graphite samples prepared using this system. The results showed that the beam current of 12C- for each sample was more than 40 μA, the carbon contamination introduced during the sample preparation process was ∼ 2 × 10–15, and that the new sample preparation system is compact, low-contamination, and efficient and meets the GXNU-AMS requirements for 14C samples.

Many waterflooding oil fields, injecting water into an oil-bearing reservoir for pressure maintenance, are in their middle to late stages of development. To explore the geological conditions and improve oilfield recovery of the most important well group of the Hu 136 block, located on the border areas of three provinces (Henan, Shandong, and Hebei), Zhongyuan Oilfield, Sinopec, central China, a 14C cross-well tracer monitoring technology was developed and applied in monitoring the development status and recognize the heterogeneity of oil reservoirs. The tracer response in the production well was tracked, and the water drive speed, swept volume of the injection fluid were obtained. Finally, the reservoir heterogeneity characteristics, such as the dilution coefficient, porosity, permeability, and average pore-throat radius, were fitted according to the mathematical model of the heterogeneous multi-layer inter-well theory. The 14C-AMS technique developed in this work is expected to be a potential analytical method for evaluating underground reservoir characteristics and providing crucial scientific guidance for the mid to late oil field recovery process.

The fall armyworm (Spodoptera frugiperda), a destructive pest that originated in South and North America, spread to China in early 2019. Controlling this invasive pest requires an understanding of its population structure and migration patterns, yet the invasion genetics of Chinese S. frugiperda is not clear. Here, using the mitochondrial cytochrome oxidase subunit I (COI) gene, triose phosphate isomerase (Tpi) gene and eight microsatellite loci, we investigated genetic structure and genetic diversity of 16 S. frugiperda populations in China. The Tpi locus identified most S. frugiperda populations as the corn-strains, and a few were heterozygous strains. The microsatellite loci revealed that the genetic diversity of this pest in China was lower than that in South America. Furthermore, we found moderate differentiation among the populations, distinct genetic structures between adjacent populations and abundant genetic resources in the S. frugiperda populations from China sampled across 2 years. The survival rate of S. frugiperda was significantly higher when it was fed on corn leaves than on rice leaves, and the larval stage mortality rate was the highest under both treatments. Our results showed that S. frugiperda probably invaded China via multiple independent introductions and careful pesticide control, continuous monitoring and further studies will be needed to minimize its potential future outbreak.

OBJECTIVES/GOALS: Whole-genome viral sequencing is vital to inform public health and study evolution. Arboviruses evolve in vectors, reservoir hosts, and humans, and require surveillance at all points. We developed a new rigorous method of sequencing that captures whole viral genomes in field-collected and clinical samples. METHODS/STUDY POPULATION: ClickSeq is a novel method of Next Generation Sequencing (NGS) library synthesis using azido-nucleotides to terminate reverse transcription. The cDNA generated can be ligated to sequencing and indexing primers at room temperature using copper (Cu I) and vitamin C. With this approach, we designed primers located ~250 bp apart along the genomes of the arboviruses Chikungunya 37797, Zika Dakar, Yellow Fever Asibi, Dengue serotype 2, West Nile 385-99, and St. Louis Encephalitis Virus (SLEV) clade II. We tested this method with varying viral titers: lab-infected mosquito pools, field-collected mosquito pools from a Texas West Nile and SLEV outbreak, and patient isolates from a Pakistani CHIKV outbreak. The cDNA was sequenced in the UTMB NGS Core and aligned using bowtie. RESULTS/ANTICIPATED RESULTS: The use of a single protocol to capture whole viral genomes including UTRs for multiple viruses from different sample collection styles is ideal for arboviruses. Primers for multiple viruses were pooled and used to sequence mosquito pools. The Tiled ClickSeq method captured whole viral genomes without the need for host depletion. UTRs were captured even when the viral strain used for primer design differed from the resulting strain. Discreet variants were captured in both the hypervariable nsP3 region and the UTR in the patient isolates from the CHIKV outbreak compared to the 2017 outbreak. Texas WNV and SLEV outbreaks are now defined from the 2020 outbreak and can be further tracked to update public health measures and understand viral evolution. DISCUSSION/SIGNIFICANCE: UTRs impact both human and mosquito fitness, leading to further outbreaks. Tiled ClickSeq aims to capture whole viral genomes with a method and cost that can be implemented by public health researchers to understand disease evolution as it happens to update both public health and basic virology to the effects of evolution on arboviruses.

Current studies on inhibitory effects of n-3 PUFA on pro-inflammatory cytokines have inconsistent results. Thus, a meta-analysis of randomised controlled trials was conducted to identify the effects of n-3 PUFA administration on circulating IL-6 and TNF in patients with cancer. Studies that examined the effects of n-3 PUFA administration on circulating IL-6 and TNF in patients with cancer were identified by searching PubMed and EMBASE from January 1975 to February 2021. Differences in n-3 PUFA administration and control conditions were determined by calculating standardised mean differences (SMD) with 95 % CI. Twenty studies involving 971 patients met the inclusion criteria. The overall SMD were 0·485 (95 % CI 0·087, 0·883) for IL-6 and 0·712 (95 % CI 0·461, 0·962) for TNF between n-3 PUFA administration and control conditions. Sources of heterogeneity were not found through subgroup and meta-regression analyses. Publication bias was observed in TNF with a slight contribution to the effect size. n-3 PUFA can reduce circulating IL-6 and TNF levels in patients with cancer. Results supported the recommendation of n-3 PUFA as adjuvant therapy for patients with cancer, possibly excluding head and neck cancer, owing to their anti-inflammatory properties.

The aim of this study was to determine the pregnancy loss rate of amniocentesis with double-needle insertions in twin pregnancies. This was a retrospective study of twin pregnancies who underwent amniocentesis with double-needle insertion between 2010 and 2019 at a single center. The pregnancy loss rates were recorded as single or double fetal loss before 24 weeks’ gestation and within 4 weeks after the procedure. Risk factors for pregnancy loss after amniocentesis were also assessed. A total of 678 twin pregnancies with amniocentesis were finally included. The pregnancy loss rates before 24 weeks’ gestation and within 4 weeks after the procedure were 0.9% and 1.9%, respectively. Only one fetal loss was presumed to be a direct result of the procedure. All other cases were complicated by structural or chromosomal anomalies. Twin pregnancies with abnormal ultrasound findings had a significantly higher rate of pregnancy loss with a relative risk of 4.81 (95% CI [1.03, 22.2]). Our study showed a low pregnancy loss rate after amniocentesis in twin pregnancies with double-needle insertions technique of sampling, which can help decision making in prenatal screening and diagnosis for twin pregnancies.

The Harihada–Chegendalai ophiolitic mélange, which is located between the Bainaimiao arc and the North China Craton, holds significant clues regarding the tectonic setting of the southern margin of the Central Asian Orogenic Belt. The ophiolitic mélange is mainly composed of gabbroic and serpentinized ultramafic rocks. Here, zircon U–Pb dating, in situ zircon Hf isotopic, whole-rock geochemical and in situ mineral chemical data from the ophiolitic mélange are reported. The zircons in the gabbroic rocks yielded concordia U–Pb ages of 450–448 Ma and exhibited slightly positive ϵHf(t) values (0.87–4.34). The geochemical characteristics of the gabbroic rocks indicate that they were generated from a mantle wedge metasomatized by subduction-derived melts from sediments with continental crust contamination, in a fore-arc tectonic setting. These rocks also experienced the accumulation of plagioclase. The geochemical characteristics of the ultramafic rocks and their Cr-spinels indicate that they may constitute part of residual mantle that has experienced a high degree of partial melting and has interacted with fluids/melts released from the subducted slab in the same fore-arc tectonic setting. The ophiolitic mélange may therefore have formed in this fore-arc tectonic setting, resulting from the northward subduction of the South Bainaimiao Ocean beneath the Bainaimiao arc during Late Ordovician time, prior to the collision between the Bainaimiao arc and the North China Craton during the Silurian to Carboniferous periods.

The present study evaluated effects of dietary supplementation with tryptophan (Trp) on muscle growth, protein synthesis and antioxidant capacity in hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂. Fish were fed six different diets containing 2·6 (control), 3·1, 3·7, 4·2, 4·7 and 5·6 g Trp/kg diet for 56 d, respectively. Results showed that dietary Trp significantly (1) improved muscle protein content, fibre density and frequency of fibre diameter; (2) up-regulated the mRNA levels of PCNA, myf5, MyoD1, MyoG, MRF4, IGF-I, IGF-II, IGF-IR, PIK3Ca, TOR, 4EBP1 and S6K1; (3) increased phosphorylation levels of AKT, TOR and S6K1; (4) decreased contents of MDA and PC, and increased activities of CAT, GST, GR, ASA and AHR; (5) up-regulated mRNA levels of CuZnSOD, CAT, GST, GPx, GCLC and Nrf2, and decreased Keap1 mRNA level; (6) increased nuclear Nrf2 protein level and the intranuclear antioxidant response element-binding ability, and reduced Keap1 protein level. These results indicated that dietary Trp improved muscle growth, protein synthesis as well as antioxidant capacity, which might be partly related to myogenic regulatory factors, IGF/PIK3Ca/AKT/TOR and Keap1/Nrf2 signalling pathways. Finally, based on the quadratic regression analysis of muscle protein and MDA contents, the optimal Trp requirements of hybrid catfish (21·82–39·64 g) were estimated to be 3·94 and 3·93 g Trp/kg diet (9·57 and 9·54 g/kg of dietary protein), respectively.

ABSTRACT IMPACT: Our improved understanding of the changes in chromatin accessibility that occur in persistent pain states may identify regulatory genomic elements that play essential roles in modulating gene expression in the DRG. OBJECTIVES/GOALS: Efforts to understand genetic variability involved in an individual’s susceptibility to persistent pain support a role for upstream regulation by epigenetic mechanisms. Our objective was to examine the transcriptomic and epigenetic basis of persistent pain following nerve injury. METHODS/STUDY POPULATION: We used a multiomic approach to identify novel molecular pathways associated with nerve injury-induced pain hypersensitivity. Adult Sprague Dawley rats were randomized to Chronic Constriction Injury (CCI) to the sciatic nerve or no treatment (naive). The ipsilateral L4-L6 dorsal root ganglia (DRG)s were removed on Day 14 and used for ChIP-seq for H3K4me1, ATAC-seq, and RNA-seq. We assessed for differential chromatin accessibility, transcription factor motifs, and enrichment for biological processes in chromatin accessible regions associated with cis-regulatory regions identified by ATAC-seq and H3K4me1 enrichment. Luciferase assays determined the functional significance of these sequences. RESULTS/ANTICIPATED RESULTS: We identified 58,446 genomic regions where H3K4me1 enrichment overlapped with chromatin accessibility. Differential analysis identified 2145 of these 58,446 regions that had changes in accessibility after CCI. The majority of these regions were located in introns or intergenic regions. Functional annotation of the differentially accessible regions identified disparate molecular functions enriched following nerve injury which suggests that altered chromatin structure plays a role in the development of mechanical hypersensitivity. Motif analysis identified specific transcription factor families whose binding sequences were enriched in regions of increased or decreased accessibility. Luciferase assays showed significant enhancement or repression of gene transcription. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our data provides a comprehensive map of chromatin accessibility changes in the DRG after CCI and emphasizes the importance of chromatin structure in the development and maintenance of chronic pain.

Six acidic dykes were discovered surrounding the Laiziling pluton, Xianghualing area, in the western Cathaysia Block, South China. A number of captured zircons are found in two of these acidic dykes. By detailed U–Pb dating, Lu–Hf isotopes and trace-element analysis, we find that these zircons have ages clustered at c. 2.5 Ga. Two acidic dyke samples yielded upper intersection point 206U/238Pb ages of 2505 ± 42 Ma and 2533 ± 22 Ma, and weighted mean 207Pb/206Pb ages of 2500 ± 30 Ma and 2535 ± 16 Ma. The majority of these zircons have high (Sm/La)N, Th/U and low Ce/Ce* ratios, indicating a magmatic origin, but some grains were altered by later hydrothermal fluid. Additionally, the magmatic zircons have high Y, U, heavy rare earth element, Nb and Ta contents, indicating that their host rocks were mainly mafic rocks or trondhjemite–tonalite–granodiorite rock series. Equally, their moderate Y, Yb, Th, Gd and Er contents also indicate that a mafic source formed in a continental volcanic-arc environment. These zircons have positive ϵHf(t) values (2.5–6.9) close to zircons from the depleted mantle, with TDM (2565–2741 Ma) and TDM2 (2608–2864 Ma) ages close to their formation ages, indicating that these zircons originated directly from depleted mantle magma, or juvenile crust derived from the depleted mantle in a very short period. We therefore infer that the Cathaysia Block experienced a crustal growth event at c. 2.5 Ga.

Corona Virus Disease 2019 (COVID-19) has presented an unprecedented challenge to the health-care system across the world. The current study aims to identify the determinants of illness severity of COVID-19 based on ordinal responses. A retrospective cohort of COVID-19 patients from four hospitals in three provinces in China was established, and 598 patients were included from 1 January to 8 March 2020, and divided into moderate, severe and critical illness group. Relative variables were retrieved from electronic medical records. The univariate and multivariate ordinal logistic regression models were fitted to identify the independent predictors of illness severity. The cohort included 400 (66.89%) moderate cases, 85 (14.21%) severe and 113 (18.90%) critical cases, of whom 79 died during hospitalisation as of 28 April. Patients in the age group of 70+ years (OR = 3.419, 95% CI: 1.596–7.323), age of 40–69 years (OR = 1.586, 95% CI: 0.824–3.053), hypertension (OR = 3.372, 95% CI: 2.185–5.202), ALT >50 μ/l (OR = 3.304, 95% CI: 2.107–5.180), cTnI >0.04 ng/ml (OR = 7.464, 95% CI: 4.292–12.980), myohaemoglobin>48.8 ng/ml (OR = 2.214, 95% CI: 1.42–3.453) had greater risk of developing worse severity of illness. The interval between illness onset and diagnosis (OR = 1.056, 95% CI: 1.012–1.101) and interval between illness onset and admission (OR = 1.048, 95% CI: 1.009–1.087) were independent significant predictors of illness severity. Patients of critical illness suffered from inferior survival, as compared with patients in the severe group (HR = 14.309, 95% CI: 5.585–36.659) and in the moderate group (HR = 41.021, 95% CI: 17.588–95.678). Our findings highlight that the identified determinants may help to predict the risk of developing more severe illness among COVID-19 patients and contribute to optimising arrangement of health resources.

This paper proposes a novel laser beam tracking mechanism for a mobile target robot that is used in shooting ranges. Compared with other traditional tracking mechanisms and modules, the proposed laser beam tracking mechanism is more flexible and low cost in use. The mechanical design and the working principle of the tracking module are illustrated, and the complete control system of the mobile target robot is introduced in detail. The tracking control includes two main steps: localizing the mobile target robot with regards to the position of the laser beam and tracking the laser beam by the linear quadratic regulator (LQR). First of all, the state function of the control system is built for this tracking system; second, the control law is deduced according to the discretized state function; lastly, the stability of the control method is proved by the Lyapunov theory. The experimental results demonstrate that the Hue, Saturation, Value feature-extracting method is robust and is qualified to be used for localization in the laser beam tracking control. It is verified through experiments that the LQR method is of better performance than the conventional Proportional Derivative control in the aspect of converge time, lateral error control, and distance error control.