Metabolomics integrated with transcriptomics reveals the changes during developmental stages in Shiraia bambusicola.

Shiraia bambusicola is a fungus with high economic value widely used in medicine, agriculture, and food. We wished to understand the genes and metabolites changes involved in the different developmental stages of S. bambusicola. So, to reveal key genes and metabolites in the main active metabolite, the  were analyzed in different developmental stages of S. bambusicola fruiting body. A total of 29,137 Unigenes were annotated. In the whole growth process, differentially expressed genes were involved in the pathways of cytochrome P450, transcription factors, transporters, and so on, while in the early stage of growth, genes enriching to synthesis pathways of basic substances. In the middle stage of growth, genes with more prominent changes were involved in the pathways of the cell cycle, cancer mechanisms, and aminobenzoate degradation; in the later stage of growth, differentially expressed genes that enriched synthesis pathways of secondary metabolites. A total of 612 metabolites were detected from different growth stages of S. bambusicola. Among them, coumarins, alkaloids, rutin, liquiritigenin, quercetin, and other medically relevant metabolites were detected for the first time. We have identified 31 secondary metabolites,  relevantly only accumulated in the early and middle stage, but not detected in the later stage, such as flavonols, coumarins, nucleotides and its derivates and hydroxycinnamoyl derivatives. The differential genes and metabolites of the same group were enriched in 127 pathways, and more significantly in ubiquinone and other terpenoid quinone biosynthesis, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and phenylpropanoid biosynthesis. The correlation networks of several significantly enriched pathways were analyzed, and the relationships within and between these pathways, genes, and metabolites, were analyzed. The synthetic pathway of hypocrellin has been speculated upon. We believe that hypocrellin is synthesized in S. bambusicola via the shikimic acid pathway followed by phenylalanine, tyrosine, and tryptophan biosynthesis pathway, then the ubiquinone and other terpenoid quinone biosynthesis pathway, and finally a series of polymerization and modification reactions. Several genes and metabolites involved in the biosynthesis of hypocrellin have been identified. This study provides a reference for further research on S. bambusicola, by providing a basis for its use and development.

Development of a simplified RT-PCR without RNA isolation for rapid detection of RNA viruses in a single small brown planthopper (Laodelphax striatellus Fallén).

BACKGROUND: The small brown planthopper (SBPH) is an important pest of cereal crops and acts as a transmission vector for multiple RNA viruses. Rapid diagnosis of virus in the vector is crucial for efficient forecast and control of viral disease. Reverse transcription polymerase chain reaction (RT-PCR) is a rapid, sensitive and reliable method for virus detection. The traditional RT-PCR contains a RNA isolation step and is widely used for virus detection in insect. However, using the traditional RT-PCR for detecting RNA virus in individual SBPHs becomes challenging because of the expensive reagents and laborious procedure associated with RNA isolation when processing a large number of samples. RESULTS: We established a simplified RT-PCR method without RNA isolation for RNA virus detection in a single SBPH. This method is achieved by grinding a single SBPH in sterile water and using the crude extract directly as the template for RT-PCR. The crude extract containing the virus RNA can be prepared in approximately two minutes. Rice stripe virus (RSV), rice black streaked dwarf virus (RBSDV) and Himetobi P virus (HiPV) were successfully detected using this simplified method. The detection results were validated by sequencing and dot immunobinding assay, indicating that this simplified method is reliable for detecting different viruses in insects. The evaluation of the sensitivity of this method showed that both RSV and HiPV can be detected when the cDNA from the crude extract was diluted up to 103 fold. Compared to the traditional RT-PCR with RNA isolation, the simplified RT-PCR method greatly reduces the sample processing time, decreases the detection cost, and improves the efficiency by avoiding RNA isolation. CONCLUSIONS: A simplified RT-PCR method is developed for rapid detection of RNA virus in a single SBPH without the laborious RNA isolation step. It offers a convenient alternative to the traditional RT-PCR method.

HPV prevalence and genotype distribution among 38 056 women in Weifang, China: a cross-sectional study.

OBJECTIVES: Human papillomavirus (HPV) infection is closely associated with cervical cancer, especially the persistent infection of high-risk HPV (HR-HPV) genotypes. Therefore, investigating the HPV prevalence, age-specific, genotype distribution and the impact of the COVID-19 pandemic among large populations was essential for HPV screening and optimising vaccination. DESIGN: This was a cross-sectional study. METHODS: A total of 38 056 cervical epithelial cell specimens were collected in Weifang city from January 2018 to December 2022. The study was divided into seven age groups based on the age of the participants. HPV genotype testing was performed by using a commercial kit which is designed for the detection of 23 HPV genotypes. RESULT: A total of 8998 women were infected with HPV, with an overall positive rate of 23.64% (8998/38 056). Single infection of HPV was dominant among different age groups, which accounted for 71.33% of total infections. The most prevalent genotype was HR-HPV 16 (4.33%), followed by 52, 58, 53 and 68. Low-risk HPV (LR-HPV) 42 exhibited the highest prevalence (2.19%) among six LR-HPV genotypes, representing a novel finding. There was a significant difference in the prevalence across different age groups (p<0.01), with the highest prevalence in the group under 25 years old. During the 3 year COVID-19 breakout period, the number of HPV samples received in 2020, 2021 and 2022 was reduced by 24.03%, 14.79% and 24.76%, respectively. In 2018-2022, the annual prevalence varied between 21.09% and 25.30%, with a decreasing trend, while the prevalence of HR-HPV 39, 56, 31 and LR-HPV 42 increased. CONCLUSION: This study indicates a high-HPV infection rate and age-specific distribution characteristics of HPV genotype infections, as well as analyses of the impact of the COVID-19 outbreak on the HPV prevalence, which provides an epidemiological basis for the control and prevention of HPV infection in this region.

School Bullying Victimization and Associated Factors Among School-Aged Adolescents in China.

There has been a significant amount of research on correlates of bullying victimization, but most prior studies are descriptive and do not distinguish between different types of bullying. The current study used a case-control study design to explore factors related to different types of bullying victimization, including physical, relational, verbal, sexual, property, and poly-bullying victimization. This study was conducted in a southern city in China, including 3054 cases who self-reported being victims of school bullying and 3054 controls who reported not being involved in any school bullying in the past 12 months. Each victim case was matched with a control on gender, school, and grade level. Univariate logistic analyses and multivariate conditional logistic regression analyses were used to identify factors associated with being a victim of school bullying. Results suggest physical bullying victimization was only associated with a family-level characteristic (parenting style) while the other four types of bullying victimization (relational, verbal, sexual, and property bullying) and poly-bullying victimization were associated with multiple social domain variables at individual, family, and school levels. Findings from this study provide evidence of factors for different types of bullying victimization and have implications for potential measures to prevent bullying. Measures from multiple social domains, including individual, family and school (e.g., developing healthy behaviors, improving social skills, positive parent-child interactions, building trust between teachers and peers, and forming strong friendships), should be considered in order to effectively prevent adolescent victimization from bullying.

Changes in microbial community and enzyme activity in soil under continuous pepper cropping in response to Trichoderma hamatum MHT1134 application.

To clarify the control effects of Trichoderma hamatum strain MHT1134 on Fusarium wilt in continuous pepper cropping fields and its regulatory effects on soil microecology, the physical and chemical properties, enzyme activities, community structures of soil samples from five field types were analysed. Samples were taken from fields that had been continuously planted for 1, 5, 9 years, and applied the strain MHT1134 for 1 and 2 years. The MHT1134 control effects on pepper wilt after application 1 year and 2 years were 63.03% and 70.21%, respectively. 4 kinds of physical and chemical indexes and 6 kinds of enzyme activities in soil were increased. With the continuous cropping years increased, the microbial abundance and diversity decreasing significantly. The relative abundances of Fusarium, Gibberella increased along with the planting years, but decreased after the MHT11134 application. However, the relative abundances of Trichoderma and Chaetomium significantly increased. Additionally, as the cropping years increased, the soil abundance of Actinobacteria gradually decreased, but it significantly increased from 17.56 to 22.44% after the MHT1134 application. Thus, strain MHT1134 effectively improved the microbial community structure of the soil, and it also positively affected soil quality. A continuous application may improve the control effect.

Revised road traffic safety law and years of life lost due to traffic deaths in China, 2002-2019.

Legal intervention is a powerful tool to reduce road traffic injuries (RTIs). China amended the Road Traffic Safety Law in 2011, but the impact of amended law on traffic crash deaths is still unknown. In this study, we conducted an interrupted time series analysis and examined years of life lost (YLLs) per 100,000 population as the assessment indicator to evaluate the association of road traffic safety law and traffic crash mortality. Annual YLLs data due to traffic deaths from 2002 to 2019 in China were obtained from the Global Burden of Disease (GBD) 2019. After implementation of the revised law, the average level of total YLLs per 100,000 population due to traffic deaths decreased from 1133.14 to 848.87, and the slope of annual YLLs per 100,000 population decreased by 30.11 (95% CI: 22.46, 37.75), indicating a steeper downward trend. The revised traffic law was associated with YLLs reduction due to traffic deaths for males, females, all age groups, pedestrians, motor vehicle users, and other road users, as well as traffic deaths attributed to alcohol use and tobacco use. These findings suggested that the revised Road Traffic Safety Law improved road safety by decreasing YLLs due to traffic deaths in China. However, the burden of RTIs is still heavy and efforts to further improve traffic laws and the adoption of other interventions are urgently needed.

Sequencing and Functional Annotation of the Whole Genome of Shiraia bambusicola.

Shiraia bambusicola is a rare medicinal fungus found in China that causes bamboo plants to decay and die with severe infection. Hypocrellin, its main active ingredient, is widely used in several fields, such as medicine, agriculture, and food industry. In this study, to clarify the genomic components, taxonomic status, pathogenic genes, secondary metabolite synthesis pathways, and regulatory mechanisms of S. bambusicola, whole-genome sequencing, assembly, and functional annotation were performed using high-throughput sequencing and bioinformatics approaches. It was observed that S. bambusicola has 33 Mb genome size, 48.89% GC content, 333 scaffolds, 2590 contigs, 10,703 genes, 82 tRNAs, and 21 rRNAs. The total length of the repeat sequence is 2,151,640 bp. The annotation of 5945 proteins was obtained from InterProScan hits based on the Gene Ontology database. Phylogenetic analysis showed that S. bambusicola belongs to Shiraiaceae, a new family of Pleosporales. It was speculated that there are more than two species or genus in Shiraiaceae. According to the annotation, 777 secreted proteins were associated with virulence or detoxification, including 777 predicted by the PHI database, 776 by the CAZY and Fungal CytochromeP450 database, and 441 by the Proteases database. The 252 genes associated with the secondary metabolism of S. bambusicola were screened and enriched into 28 pathways, among which the terpenoids, staurosporine, aflatoxin, and folate synthesis pathways have not been reported in S. bambusicola The T1PKS was the main gene cluster among the 28 secondary metabolite synthesis gene clusters in S. bambusicola The analysis of the T3PKS gene cluster related to the synthesis of hypocrellin showed that there was some similarity between S. bambusicola and 10 other species of fungi; however, the similarity was very low wherein the highest similarity was 17%. The genomic information of S. bambusicola obtained in this study was valuable to understand its genetic function and pathogenicity. The genomic information revealed that several enzyme genes and secreted proteins might be related to their host interactions and pathogenicity. The annotation and analysis of its secondary metabolite synthesis genes and gene clusters will be an important reference for future studies on the biosynthesis and regulation mechanism of the secondary metabolites, contributing to the discovery of new metabolites and accelerating drug development and application.