An engineered Pichia pastoris platform for the biosynthesis of silk-based nanomaterials with therapeutic potential.

Silk fibroin (SF) from the cocoon of silkworm has exceptional mechanical properties and biocompatibility and is used as a biomaterial in a variety of fields. Sustainable, affordable, and scalable manufacturing of SF would enable its large-scale use. We report for the first time the high-level secretory production of recombinant SF peptides in engineered Pichia pastoris cell factories and the processing thereof to nanomaterials. Two SF peptides (BmSPR3 and BmSPR4) were synthesized and secreted by P. pastoris using signal peptides and appropriate spacing between hydrophilic sequences. By strain engineering to reduce protein degradation, increase glycyl-tRNA supply, and improve protein secretion, we created the optimized P. pastoris chassis PPGSP-8 to produce BmSPR3 and BmSPR4. The SF fed-batch fermentation titers of the resulting two P. pastoris cell factories were 11.39 and 9.48 g/L, respectively. Protein self-assembly was inhibited by adding Tween 80 to the medium. Recombinant SF peptides were processed to nanoparticles (NPs) and nanofibrils. The physicochemical properties of nanoparticles R3NPs and R4NPs from the recombinant SFs synthesized in P. pastoris cell factories were similar or superior to those of RSFNPs (Regenerated Silk Fibroin NanoParticles) originating from commercially available SF. Our work will facilitate the production by microbial fermentation of functional SF for use as a biomaterial.

Microstructure, Tensile Properties, and Fracture Toughness of an In Situ Rolling Hybrid with Wire Arc Additive Manufacturing AerMet100 Steel.

As a type of ultra-high strength steel, AerMet100 steel is used in the aerospace and military industries. Due to the fact that AerMet100 steel is difficult to machine, people have been exploring the process of additive manufacturing to fabricate AerMet100 steel. In this study, AerMet100 steel was produced using an in situ rolling hybrid with wire arc additive manufacturing. Microstructure, tensile properties, and fracture toughness of as-deposited and heat-treated AerMet100 steel were evaluated in different directions. The results reveal that the manufacturing process leads to grain fragmentation and obvious microstructural refinement of the AerMet100 steel, and weakens the anisotropy of the mechanical properties. After heat treatment, the microstructure of the AerMet100 steel is mainly composed of lath martensite and reversed austenite. Alloy carbides are precipitated within the martensitic matrix, and a high density of dislocations is the primary strengthening mechanism. The existence of film-like austenite among the martensite matrix enhances the toughness of AerMet100 steel, which coordinates stress distribution and restrains crack propagation, resulting in an excellent balance between strength and toughness. The AerMet100 steel with in situ rolling is isotropy and achieves the following values: an average ultimate strength of 1747.7 ± 16.3 MPa, yield strength of 1615 ± 40.6 MPa, elongation of 8.3 ± 0.2% in deposition direction, and corresponding values in the building direction are 1821.3 ± 22.1 MPa, 1624 ± 84.5 MPa, and 7.6 ± 1.7%, and the KIC value up to 70.6 MPa/m0.5.

The protein methyltransferase TrSAM inhibits cellulase gene expression by interacting with the negative regulator ACE1 in Trichoderma reesei.

Protein methylation is a commonly posttranslational modification of transcriptional regulators to fine-tune protein function, however, whether this regulation strategy participates in the regulation of lignocellulase synthesis and secretion in Trichoderma reesei remains unexplored. Here, a putative protein methyltransferase (TrSAM) is screened from a T. reesei mutant with the ability to express heterologous ß-glucosidase efficiently even under glucose repression. The deletion of its encoding gene trsam causes a significant increase of cellulase activities in all tested T. reesei strains, including transformants of expressing heterologous genes using cbh1 promotor. Further investigation confirms that TrSAM interacts with the cellulase negative regulator ACE1 via its amino acid residue Arg383, which causes a decrease in the ACE1-DNA binding affinity. The enzyme activity of a T. reesei strain harboring ACE1R383Q increases by 85.8%, whereas that of the strains with trsam or ace1 deletion increases by more than 100%. By contrast, the strain with ACE1R383K shows no difference to the parent strain. Taken together, our results demonstrate that TrSAM plays an important role in regulating the expression of cellulase and heterologous proteins initiated by cbh1 promotor through interacting with ACE1R383. Elimination and mutation of TrSAM and its downstream ACE1 alleviate the carbon catabolite repression (CCR) in expressing cellulase and heterologous protein in varying degrees. This provides a new solution for the exquisite modification of T. reesei chassis.

Stigma and its influencing factors for seeking professional psychological help among pregnant women: A cross-sectional study.

BACKGROUND: Although the stigma associated with seeking professional psychological help has received significant attention, few studies have focused on the status and factors influencing this stigma among pregnant women, especially those with a Chinese cultural background. This study aimed to assess the status of stigma associated with seeking professional psychological help and its influencing factors. METHODS: A total of 1325 pregnant women were recruited from Hunan Province, China. The Stigma for Seeking Professional Psychological Help questionnaire, Patient Health Questionnaire-9, Perceived Social Support Scale, Simplified Coping Style Questionnaire, and questionnaires on sociodemographic characteristics and pregnancy-related information were used. Descriptive, univariate, correlation, and multivariate linear regression analyses were used to investigate the status of stigma and identify its influencing factors. FINDINGS: Pregnant Chinese women experienced a mild to moderate level of stigma associated with seeking professional psychological help, and self-stigma was more common than social stigma associated with seeking psychological help. The results of the multivariate analysis suggested that perceived social support, positive coping styles, negative coping styles, and depressive symptoms were the factors influencing self-stigma and social stigma associated with seeking professional psychological help. Advanced age and a low educational level were the only predictors associated with self-stigma, while a history of multiple pregnancies had an effect only on social stigma associated with seeking professional psychological help. CONCLUSION: Chinese pregnant women experienced a mild to moderate level of stigma associated with seeking professional psychological help. Destigmatizing interventions focusing on self-stigma and social stigma should be designed according to maternal characteristics, social support, coping style, and depressive symptoms.

Women's experience of polycystic ovary syndrome management: A systematic review and meta-synthesis.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common chronic condition in women of child-bearing age. There is currently no effective treatment, so early and long-term management is essential. However, there are many problems in the practice of disease management in women with PCOS that make it difficult to achieve good outcomes. OBJECTIVE: To explore women's experience of PCOS management and identify the relevant facilitators and barriers to management. SEARCH STRATEGY: A structured search was undertaken in five bibliographic databases (MEDLINE, Web of Science, CINAHL, Embase, PubMed, and Cochrane) from the date of establishment of the database up to December 2022. SELECTION CRITERIA: All qualitative and mixed-methods studies available in English describing the experience of PCOS management from the patients' perspective were included. DATA COLLECTION AND ANALYSIS: The Joanna Briggs Institute Qualitative Assessment and Review Instrument was used to appraise study quality. The evidence was synthesized using a pragmatic meta-aggregative approach guided by the capability, opportunity, and motivation model of behavior (COM-B). MAIN RESULTS: A total of 13 studies were included with 85 equivocal findings and 12 credible findings. The findings were meta-aggregated into three themes: (1) capability of women with PCOS, including patients' attitudes toward disease and management, knowledge, and skills of the disease; (2) opportunities in PCOS management, including information about PCOS, diagnostic delay, disease characteristics, disease management plan, and logistical and environmental problems; and (3) motivation in PCOS management, including impact of symptoms, perceived needs, support and feedback, and unpleasant medical experience. CONCLUSIONS: This study identifies facilitators and barriers to PCOS management from the patient perspective, which can guide the design and implementation of PCOS management programs for patients. This study also provides information for future research into how the COM-B theory can be incorporated into specific management plans to promote patient action.

Trajectories of perinatal suicidal ideation from early pregnancy to six weeks postpartum and their influencing factors: A prospective longitudinal study.

Few studies focused on the trajectory of perinatal suicidal ideation from early pregnancy and covered whole routine perinatal periods. This study aimed to investigate the prevalence of suicidal ideation from early pregnancy to six weeks postpartum, and further explore the trajectories of perinatal suicidal ideation and their risk factors. A prospective longitudinal study was conducted in a comprehensive tertiary hospital in Hunan province, China among 1089 participants. Perinatal suicidal ideation and depression were assessed by item 9 of Patient Health Questionnaire 9 and the remaining eight items. Sociodemographic and psychological factors were collected by self-reported comprehensive questionnaires. Latent Growth Curve and Growth Mixture Modeling were used to identify the trajectories of suicidal ideation and logistic regression was used to explore risk factors of trajectories. A total of 629 participants were included. The prevalence of suicidal ideation was 16.4% from early pregnancy to six weeks postpartum, with 12.1% in pregnancy and 7.8% in postpartum. Two trajectories were identified: "persistent low levels of suicidal ideation" (92.9%) and "persistent high levels of suicidal ideation" (7.1%). 40.63% of women who screened positive for suicidal ideation during early pregnancy were in the "persistent high level of suicidal ideation" trajectory. A low level of neuroticism and anxiety symptoms during early pregnancy was associated with "persistent high levels of suicidal ideation". In conclusion, suicidal ideation of women during the perinatal period was dynamic, Suicidal ideation screening and identification in early pregnancy should be addressed for perinatal women to facilitate timely early interventions.

Multi-omics strategies uncover the molecular mechanisms of nitrogen, phosphorus and potassium deficiency responses in Brassica napus.

BACKGROUND: Nitrogen (N), phosphorus (P) and potassium (K) are critical macronutrients in crops, such that deficiency in any of N, P or K has substantial effects on crop growth. However, the specific commonalities of plant responses to different macronutrient deficiencies remain largely unknown. METHODS: Here, we assessed the phenotypic and physiological performances along with whole transcriptome and metabolomic profiles of rapeseed seedlings exposed to N, P and K deficiency stresses. RESULTS: Quantities of reactive oxygen species were significantly increased by all macronutrient deficiencies. N and K deficiencies resulted in more severe root development responses than P deficiency, as well as greater chlorophyll content reduction in leaves (associated with disrupted chloroplast structure). Transcriptome and metabolome analyses validated the macronutrient-specific responses, with more pronounced effects of N and P deficiencies on mRNAs, microRNAs (miRNAs), circular RNAs (circRNAs) and metabolites relative to K deficiency. Tissue-specific responses also occurred, with greater effects of macronutrient deficiencies on roots compared with shoots. We further uncovered a set of common responders with simultaneous roles in all three macronutrient deficiencies, including 112 mRNAs and 10 miRNAs involved in hormonal signaling, ion transport and oxidative stress in the root, and 33 mRNAs and 6 miRNAs with roles in abiotic stress response and photosynthesis in the shoot. 27 and seven common miRNA-mRNA pairs with role in miRNA-mediated regulation of oxidoreduction processes and ion transmembrane transport were identified in all three macronutrient deficiencies. No circRNA was responsive to three macronutrient deficiency stresses, but two common circRNAs were identified for two macronutrient deficiencies. Combined analysis of circRNAs, miRNAs and mRNAs suggested that two circRNAs act as decoys for miR156 and participate in oxidoreduction processes and transmembrane transport in both N- and P-deprived roots. Simultaneously, dramatic alterations of metabolites also occurred. Associations of RNAs with metabolites were observed, and suggested potential positive regulatory roles for tricarboxylic acids, azoles, carbohydrates, sterols and auxins, and negative regulatory roles for aromatic and aspartate amino acids, glucosamine-containing compounds, cinnamic acid, and nicotianamine in plant adaptation to macronutrient deficiency. CONCLUSIONS: Our findings revealed strategies to rescue rapeseed from macronutrient deficiency stress, including reducing the expression of non-essential genes and activating or enhancing the expression of anti-stress genes, aided by plant hormones, ion transporters and stress responders. The common responders to different macronutrient deficiencies identified could be targeted to enhance nutrient use efficiency in rapeseed.

Research progress and applications of colorful Brassica crops.

MAIN CONCLUSION: We review the application and the molecular regulation of anthocyanins in colorful Brassica crops, the creation of new germplasm resources, and the development and utilization of colorful Brassica crops. Brassica crops are widely cultivated: these include oilseed crops, such as rapeseed, mustards, and root, leaf, and stem vegetable types, such as turnips, cabbages, broccoli, and cauliflowers. Colorful variants exist of these crop species, and asides from increased aesthetic appeal, these may also offer advantages in terms of nutritional content and improved stress resistances. This review provides a comprehensive overview of pigmentation in Brassica as a reference for the selection and breeding of new colorful Brassica varieties for multiple end uses. We summarize the function and molecular regulation of anthocyanins in Brassica crops, the creation of new colorful germplasm resources via different breeding methods, and the development and multifunctional utilization of colorful Brassica crop types.

Mapping and Identifying Candidate Genes Enabling Cadmium Accumulation in Brassica napus Revealed by Combined BSA-Seq and RNA-Seq Analysis.

Rapeseed has the ability to absorb cadmium in the roots and transfer it to aboveground organs, making it a potential species for remediating soil cadmium (Cd) pollution. However, the genetic and molecular mechanisms underlying this phenomenon in rapeseed are still unclear. In this study, a 'cadmium-enriched' parent, 'P1', with high cadmium transport and accumulation in the shoot (cadmium root: shoot transfer ratio of 153.75%), and a low-cadmium-accumulation parent, 'P2', (with a cadmium transfer ratio of 48.72%) were assessed for Cd concentration using inductively coupled plasma mass spectrometry (ICP-MS). An F2 genetic population was constructed by crossing 'P1' with 'P2' to map QTL intervals and underlying genes associated with cadmium enrichment. Fifty extremely cadmium-enriched F2 individuals and fifty extremely low-accumulation F2 individuals were selected based on cadmium content and cadmium transfer ratio and used for bulk segregant analysis (BSA) in combination with whole genome resequencing. This generated a total of 3,660,999 SNPs and 787,034 InDels between these two segregated phenotypic groups. Based on the delta SNP index (the difference in SNP frequency between the two bulked pools), nine candidate Quantitative trait loci (QTLs) from five chromosomes were identified, and four intervals were validated. RNA sequencing of 'P1' and 'P2' in response to cadmium was also performed and identified 3502 differentially expressed genes (DEGs) between 'P1' and 'P2' under Cd treatment. Finally, 32 candidate DEGs were identified within 9 significant mapping intervals, including genes encoding a glutathione S-transferase (GST), a molecular chaperone (DnaJ), and a phosphoglycerate kinase (PGK), among others. These genes are strong candidates for playing an active role in helping rapeseed cope with cadmium stress. Therefore, this study not only sheds new light on the molecular mechanisms of Cd accumulation in rapeseed but could also be useful for rapeseed breeding programs targeting this trait.

Barriers and facilitators of psychological help-seeking behaviors for perinatal women with depressive symptoms: A qualitative systematic review based on the Consolidated Framework for Implementation Research.

OBJECTIVE: To identify barriers and facilitators related to psychological help-seeking behaviors of perinatal depression from all related stakeholders (e.g., perinatal women, family members, mental health care providers, and policymakers). DESIGN: A literature search of six English-language databases (PubMed, Web of Science, Embase, PsycINFO, the Cochrane Library, CINAHL) and three Chinese-language databases (China National Knowledge Infrastructure, Wan Fang, Chinese Biomedical Literature Databases). Studies published in English or Chinese using qualitative or mixed methods to explore the psychological help-seeking behaviors of women with perinatal depression were included. Data extraction was synthesized for common themes based on the Consolidated Framework for Implementation Research. The Joanna Briggs Institute Qualitative Assessment and Review Instrument was used to appraise methodologic quality. PARTICIPANTS AND SETTINGS: Perinatal women with depression, mental health care providers (e.g., pediatricians/nurses, social workers, nurse-midwives, perinatal psychiatrists, community health workers, and administrators), partners and informal caregivers (e.g., community birth attendants, elderly mothers, and men of reproductive age) based in high, middle and low income countries. FINDINGS: Forty-three articles were included in this review and presented according to the Consolidated Framework for Implementation Research domains (in parentheses). The most common barriers to help seeking were stigma (individual characteristics), misconceptions (individual characteristics), cultural beliefs (inner setting), and lack of social support (outer setting). The most common facilitators were providing adequate support (outer setting) and perinatal health care professional training on how to detect, manage and discuss depression; establishing supportive relationships with mental health care providers; and eroding stigma (all three implementation processes). KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: This systematic review could serve as a reference framework for health authorities to develop diverse strategies for improving the psychological help-seeking behaviors of women with perinatal depression. More high-quality studies focused on the Consolidated Framework for Implementation Research characteristics of available interventions, and implementation processes are needed in future research.

Help-Seeking Intentions for Depression and Associated Factors among Chinese Perinatal Women: A Cross-Sectional Study.

A low help-seeking intention for depression is an important reason for the low number of women with perinatal depression who have sought professional help. However, evidence of help-seeking intentions for depression is still lacking in Chinese perinatal women. We aimed to investigate the help-seeking intention for depression and its associated factors among Chinese perinatal women. Participants were recruited from three comprehensive hospitals in Changsha. A total of 874 perinatal women were included in the study. The score for the help-seeking intention for depression in Chinese perinatal women was 3.65 ± 0.79, with about half of participants (58.3%) reporting that they were "likely" and "strongly likely" to seek professional help if they suffered from depression during the perinatal period. Favorable help-seeking attitudes and sufficient knowledge of mental illness help-seeking resources were positively associated with help-seeking intentions for depression. However, self-stigma decreased the help-seeking intention for depression. Chinese perinatal women had relatively positive help-seeking intentions for depression. Reducing the stigma of mental illness and help-seeking, enhancing mental health literacy, and improving attitudes toward professional psychological help-seeking of perinatal women may be the potential key components of interventions to encourage perinatal women to actively seek professional psychological help.

Attitudes toward seeking professional psychological help among Chinese pregnant women: A cross-sectional study.

BACKGROUND: There exists a gap between the high prevalence of mental health problems and negative attitudes toward seeking professional psychological help among Chinese pregnant women. This study aimed to investigate the attitudes and factors toward seeking professional psychological help among Chinese pregnant women to provide some suggestions for improving pregnant women's psychological help-seeking behaviors. METHODS: A total of 1024 pregnant women were recruited. The Chinese version of the Attitude Toward Seeking Professional Psychological Help Scale (ATSPPH) was used to assess the attitudes toward professional psychological help-seeking. Demographic and socio-psychological factors were collected by self-developed questionnaire, Perceived Social Support Scale, Simplified Coping Style Questionnaire, and Patient Health Questionnaire-9 Scale. RESULTS: 1006 pregnant women were included in the final analysis. Only 3.7 % of participants chose professionals as their first choice for psychological help-seeking. The attitudes toward seeking professional psychological help were relatively positive, with the highest score of stigma tolerance among four subscales in ATSPPH. Education level, perceived social support, and positive coping style had a positive predictive effect on the four subscales of ATSPPH. Women with prenatal depressive symptoms had a lower score in the subscales of recognition of need for psychotherapeutic help and confidence in mental health practitioner; women with higher scores of negative coping style had a lower score in the subscales of stigma tolerance and interpersonal openness. History of professional psychological help-seeking behaviors was only associated with the subscale of recognition of need for psychotherapeutic help; Maternal age was only associated with the subscale of interpersonal openness. CONCLUSION: The attitudes toward seeking professional psychological help were relatively positive among Chinese pregnant women, and stigma tolerance was the significant barrier to professional psychological help-seeking. More attention should be paid to those women with older age, negative coping style, and women with prenatal depressive symptoms.

Efficient genome editing in Claviceps purpurea using a CRISPR/Cas9 ribonucleoprotein method.

Claviceps purpurea produces many pharmacologically important ergot alkaloids (EAS), which are widely used to treat migraine and hypertension and to aid childbirth. Although an EAS biosynthetic cluster of C. purpurea has been discovered more than 20 years ago, the complete biosynthetic pathway of EAS has not been fully characterized until now. The main obstacle to elucidating this pathway and strain modification is the lack of efficient genome-editing tools for C. purpurea. The conventional gene manipulation method for C. purpurea relies on homologous recombination (HR), although the efficiency of HR in C. purpurea is very low (∼1-5%). Consequently, the disruption of target genes is laborious and time-consuming. Although CRISPR/Cas9 genome-editing methods based on in vivo Cas9 expression and gRNA transcription have been reported recently, their gene-disruption efficiency is still very low. Here, we developed an efficient genome-editing system in C. purpurea based on in vitro assembled CRISPR/Cas9 gRNA ribonucleoprotein complexes. As proof of principle, three target genes were efficiently knocked out using this CRISPR/Cas9 ribonucleoprotein complex-mediated HR system, with editing efficiencies ranging from 50% to 100%. Inactivation of the three genes, which are closely related to uridine biosynthesis (ura5), hypha morphology (rac), and EAS production (easA), resulted in a uridine auxotrophic mutant, a mutant with a drastically different phenotype in axenic culture, and a mutant that did not produce EAS, respectively. Our ribonucleoprotein-based genome-editing system has a great advantage over conventional and in vivo CRISPR/Cas9 methods for genome editing in C. purpurea, which will greatly facilitate elucidation of the EAS biosynthetic pathway and other future basic and applied research on C. purpurea.

Prevalence of suicidal ideation in pregnancy and the postpartum: A systematic review and meta-analysis.

BACKGROUND: Suicidal ideation, a significant risk factor of suicide, is considered a potential trigger for intervention prior to self-harm or suicide; however, the prevalence of maternal suicidal ideation varied widely between studies. This review aims to synthesis the available evidence to estimate the prevalence of maternal suicidal ideation before and after pregnancy. METHODS: We searched six English databases (PubMed, Web of Science, Embase, the Cochrane Library, PsycINFO, CINAHL) and three Chinese databases (China National Knowledge Infrastructure, Wang Fang, Chinese Biomedical Literature Databases) from database inceptions before August 31, 2020; and checked the reference list for relevant studies. Data in the included studies were used to calculate the prevalence of maternal suicidal ideation. Subgroup analysis and meta-regression were performed to detect the potential sources of heterogeneity. RESULTS: 6094 potentially studies were identified. 71 studies, including 23 cohort studies, 47 cross-sectional studies, and 1 RCT study, were included for final analysis. The total participants were 92146, with sample size ranged from 23 to 22118. The included studies were from 23 different countries distributing on six continents, most of the studies were conducted in Asia (n = 18), North America (n = 16), and South America (n = 15). The pooled prevalence of maternal suicidal ideation reported by eligible studies was 8% (95% CI 7-10%), with 10% in antenatal and 7% in postpartum. Prevalence significantly varied based on measuring tools, study design, study countries, and publication year, while prevalence was not conditional on the time-point assessment, sample size, and maternal age. LIMITATION: Obvious heterogeneity and no standardization tools for measuring suicidal ideation may limit the results' interpretation. CONCLUSION: Our systematic review reported the prevalence of suicidal ideation in pregnancy and postpartum was 8%. Future studies should establish standardization tools for measuring suicidal ideation and identify strategies of prevention and treatment.

Building a Versatile Protein Production Platform Using Engineered Trichoderma reesei.

Trichoderma reesei has an extremely high capacity for synthesizing and secreting proteins, thus exhibiting promise as an expression platform for heterologous proteins. However, T. reesei secretes large amounts of native proteins, which hinders its widespread application for heterologous protein production. Here, we designed and built a series of T. reesei chassis using an iterative gene deletion approach based on an efficient genome editing system. Donor DNAs with specially designed construct facilitated screening of positive deletion strains without ectopic insertion. Finally, marker-free T. reesei chassis with lower rates of native protein secretion and low levels of extracellular protease activity were constructed after 11 consecutive rounds of gene deletion. Higher production levels of three heterologous proteins─a bacterial xylanase XYL7, a fungal immunomodulatory protein LZ8, and the human serum albumin HSA─were achieved with these chassis using the cbh1 promoter. It is possible that diverse high-value proteins might be produced at a high yield using this engineered platform.

Alleviating product inhibition of Trichoderma reesei cellulase complex with a product-activated mushroom endoglucanase.

Product inhibition of cellulase is a challenging issue in industrial processes. Here, we introduced a product-activated mushroom cellulase, PaCel3A from Polyporus arcularius, into Trichoderma reesei. The filter paper activity, carboxymethyl cellulase activity, and saccharification efficiency (substrate: pretreated rice straw, PRS) of transformants increased significantly with this enzyme (by 18.4-26.8%, 13.8-22.8%, and 17.0%, respectively). A mutant of PaCel3A, PaCel3AM, obtained based on B-factor analysis, saturated mutagenesis, and residual activity assay, showed improved thermostability. The PRS saccharification efficiency using the cellulase complex from T. reesei transformants overexpressing pacel3am increased by 56.4%-63.0%. In addition, the T. reesei cellulase complex obtained by adding the purified recombinant PaCel3AM from T. reesei (rCel3aM-tr) to hydrolyze PRS resulted in increased reducing sugar yields at all sampling points, outperforming the cellulase complexes without rCel3aM-tr. These results suggest that introducing product-activated cellulase genes is a simple and feasible method to alleviate the product inhibition of cellulase.

Efficient genome editing in filamentous fungi via an improved CRISPR-Cas9 ribonucleoprotein method facilitated by chemical reagents.

DNA double-strand break (DSB) repair induced by the RNA-programmed nuclease Cas9 has become a popular method for genome editing. Direct genome editing via Cas9-CRISPR gRNA (guide RNA) ribonucleoprotein (RNP) complexes assembled in vitro has also been successful in some fungi. However, the efficiency of direct RNP transformation into fungal protoplasts is currently too low. Here, we report an optimized genome editing approach for filamentous fungi based on RNPs facilitated by adding chemical reagents. We increased the transformation efficiency of RNPs significantly by adding Triton X-100 and prolonging the incubation time, and the editing efficiency reached 100% in Trichoderma reesei and Cordyceps militaris. The optimized RNP-based method also achieved efficient (56.52%) homologous recombination integration with short homology arms (20 bp) and gene disruption (7.37%) that excludes any foreign DNA (selection marker) in T. reesei. In particular, after adding reagents related to mitosis and cell division, the further optimized protocol showed an increased ratio of edited homokaryotic transformants (from 0% to 40.0% for inositol and 71.43% for benomyl) from Aspergillus oryzae, which contains multinucleate spores and protoplasts. Furthermore, the multi-target engineering efficiency of the optimized RNP transformation method was similar to those of methods based on in vivo expression of Cas9. This newly established genome editing system based on RNPs may be widely applicable to construction of genome-edited fungi for the food and medical industries, and has good prospects for commercialization.

Risk prediction for postpartum depression based on random forest. / 随机森林算法在产后抑郁风险预测中的应用.

OBJECTIVES: To explore the application of random forest algorithm in screening the risk factors and predictive values for postpartum depression. METHODS: We recruited the participants from a tertiary hospital between June 2017 and June 2018 in Changsha City, and followed up from pregnancy up to 4-6 weeks postpartum.Demographic economics, psychosocial, biological, obstetric, and other factors were assessed at first trimesters with self-designed obstetric information questionnaire and the Chinese version of Edinburgh Postnatal Depression Scale (EPDS). During 4-6 weeks after delivery, the Chinese version of EPDS was used to score depression and self-designed questionnaire to collect data of delivery and postpartum. The data of subjects were randomly divided into the training data set and the verification data set according to the ratio of 3꞉1. The training data set was used to establish the random forest model of postpartum depression, and the verification data set was used to verify the predictive effects via the accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and AUC index. RESULTS: A total of 406 participants were in final analysis. Among them, 150 of whom had EPDS score ≥9, and the incidence of postpartum depression was 36.9%. The predictive effects of random forest model in the verification data set were at accuracy of 80.10%, sensitivity of 61.40%, specificity of 89.10%, positive predictive value of 73.00%, negative predictive value of 82.80%, and AUC index of 0.833. The top 10 predictive influential factors that screening by the variable importance measure in random forest model was antenatal depression, economic worries after delivery, work worries after delivery, free triiodothyronine in first trimesters, high-density lipoprotein in third trimester, venting temper to infants, total serum cholesterol and serum triglyceride in first trimester, hematocrit and serum triglyceride in third trimester. CONCLUSIONS: Random forest has a great advantage in risk prediction for postpartum depression. Through comprehensive evaluation mechanism, it can identify the important influential factors for postpartum depression from complex multi-factors and conduct quantitative analysis, which is of great significance to identify the key factors for postpartum depression and carry out timely and effective intervention.

Research progress in prediction of postpartum depression. / äº§åŽæŠ‘éƒé¢„æµ‹ç ”ç©¶è¿›å±•.

Postpartum depression is a kind of mental disorder caused by the comprehensive effects of economics, psychosociology, biology, obstetrics and other characteristic factors in the process of female pregnancy. The pathogenesis of postpartum depression is complicated and has not been clarified. With the process pregnancy, the influence of psychosocial and biology factors are also in dynamic change. The postpartum depression predictor among psychosocial factors are antenatal depression, life events and social supports. Among biological factors, the predictors are hypothalamic-pituitary-thyroid axis and serum lipids. Timely and effective prediction can identify the high-risk population and risk factors for postpartum depression.

MicroRNA-mRNA expression profiles and their potential role in cadmium stress response in Brassica napus.

BACKGROUND: Oilseed rape is an excellent candidate for phytoremediation of cadmium (Cd) contaminated soils given its advantages of high biomass, fast growth, moderate metal accumulation, ease of harvesting, and metal tolerance, but the cadmium response pathways in this species (Brassica napus) have yet to be fully elucidated. A combined analysis of miRNA and mRNA expression to infer Cd-induced regulation has not been reported in B. napus. RESULTS: We characterized concurrent changes in miRNA and mRNA profiles in the roots and shoots of B. napus seedlings after 10 days of 10 mg/L Cd2+ treatment. Cd treatment significantly affected the expression of 22 miRNAs belonging to 11 families in the root and 29 miRNAs belonging to 14 miRNA families in the shoot. Five miRNA families (MIR395, MIR397, MIR398, MIR408 and MIR858) and three novel miRNAs were differentially expressed in both tissues. A total of 399 differentially expressed genes (DEGs) in the root and 389 DEGs in the shoot were identified, with very little overlap between tissue types. Eight anti-regulation miRNA-mRNA interaction pairs in the root and eight in the shoot were identified in response to Cd and were involved in key plant stress response pathways: for example, four genes targeted by miR398 were involved in a pathway for detoxification of superoxide radicals. Cd stress significantly impacted the photosynthetic pathway. Transcription factor activation, antioxidant response pathways and secondary metabolic processes such as glutathione (GSH) and phenylpropanoid metabolism were identified as major components for Cd-induced response in both roots and shoots. CONCLUSIONS: Combined miRNA and mRNA profiling revealed miRNAs, genes and pathways involved in Cd response which are potentially critical for adaptation to Cd stress in B. napus. Close crosstalk between several Cd-induced miRNAs and mRNAs was identified, shedding light on possible mechanisms for response to Cd stress in underground and aboveground tissues in B. napus. The pathways, genes, and miRNAs identified here will be valuable targets for future improvement of cadmium tolerance in B. napus.