Fungal necromass is reduced by intensive drought in subsoil but not in topsoil.

The frequency and intensity of droughts worldwide are challenging the conservation of soil organic carbon (SOC) pool. Microbial necromass is a key component of SOC, but how it responds to drought at specific soil depths remains largely unknown. Here, we conducted a 3-year field experiment in a forest plantation to investigate the impacts of drought intensities under three treatments (ambient control [CK], moderate drought [30% throughfall removal], and intensive drought [50% throughfall removal]) on soil microbial necromass pools (i.e., bacterial necromass carbon, fungal necromass carbon, and total microbial necromass carbon). We showed that the effects of drought on microbial necromass depended on microbial groups, soil depth, and drought intensity. While moderate drought increased total (+9.1% ± 3.3%) and fungal (+13.5% ± 4.9%) necromass carbon in the topsoil layer (0-15 cm), intensive drought reduced total (-31.6% ± 3.7%) and fungal (-43.6% ± 4.0%) necromass in the subsoil layer (15-30 cm). In contrast, both drought treatments significantly increased the BNC in the topsoil and subsoil. Our results suggested that the effects of drought on the microbial necromass of the subsoil were more pronounced than those of the topsoil. This study highlights the complex responses of microbial necromass to drought events depending on microbial community structure, drought intensity and soil depth with global implications when forecasting carbon cycling under climate change.

Phosphorus additions imbalance terrestrial ecosystem C:N:P stoichiometry.

Carbon (C):nitrogen (N):phosphorus (P) stoichiometry in plants, soils, and microbial biomass influences productivity and nutrient cycling in terrestrial ecosystems. Anthropogenic inputs of P to ecosystems are increasing; however, our understanding of the impacts of P addition on terrestrial ecosystem C:N:P ratios remains elusive. By conducting a meta-analysis with 1413 paired observations from 121 publications, we showed that P addition significantly decreased plant, soil, and microbial biomass N:P and C:P ratios, but had negligible effects on C:N ratios. The reductions in N:P and C:P ratios became more evident as the P application rates and experimental duration increased. The P addition effects on terrestrial ecosystem C:N:P stoichiometry did not vary with ecosystem types or climates. Moreover, the responses of N:P and C:P ratios in soil and microbial biomass were associated with the responses of soil pH and fungi:bacteria ratios. Additionally, P additions increased net primary productivity, microbial biomass, soil respiration, N mineralization, and N nitrification, but decreased ammonium and nitrate contents. Decreases in plant N:P and C:P ratios were both negatively correlated to net primary productivity and soil respiration, but positively correlated to ammonium and nitrate contents; microbial biomass, soil respiration, ammonium contents, and nitrate contents all increased with declining soil N:P and C:P ratios. Our findings highlight that P additions could imbalance C:N:P stoichiometry and potentially impact the terrestrial ecosystem functions.

Application of CEEMD noise reduction algorithm in ultrasound imaging in evaluating fetuses with abnormal glucose metabolism in late pregnancy.

OBJECTIVES: To explore the predictive effect of abnormal glucose metabolism and fetal hemodynamic parameters on adverse pregnancy outcome. METHODS: One hundred and nine pregnant women with abnormal glucose metabolism during pregnancy from June 2016 to October 2018 were selected and divided into poor prognosis group (34 cases) and good prognosis group (75 cases). The hemodynamic parameters of fetal cerebral artery (MCA), umbilical artery (UA) and uterine artery of pregnancy (UT-A), including peak systolic velocity (s / D), resistance index (RI) and plasticity index (PI), were measured by color Doppler ultrasound. The receiver operating characteristic (ROC) curve of adverse pregnancy outcomes was drawn and the best threshold index was determined. RESULTS: MCA-PI poor prognosis group, MCA-RI, RI ratio (MCA/UA) are lower than the good prognosis group, Ut-A-PI is higher than the good prognosis group (P<0.05,). ROC curve analysis results show that when the MCA-PI 1.56, the sensitivity of the predicted adverse outcomes of pregnancy, the highest specificity<, was 91.18%, 80.00%, respectively. Logistic regression analysis of risk factors shows poor pregnancy outcomes include: pregnant women, older age, body mass index ≥24.0kg/m2 and a family history of diabetes. Protective factors include exercise during pregnancy, MCA-PI≥1.56, MCA-RI≥0.63 and RI The ratio (MCA/UA) ≥0.84. CONCLUSION: Color Doppler ultrasound measured MCA-PI<1.56 the most important indicators of poor pregnancy outcomes as abnormal glucose metabolism during pregnancy and predict the exact cutoff. Pregnant women, older age, body mass index ≥24.0kg/m2 and a family history of diabetes and abnormal glucose metabolism during pregnancy risk factors for adverse outcomes of pregnancy.

Integrated metabolomics and transcriptomics study of traditional herb Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao reveals global metabolic profile and novel phytochemical ingredients.

BACKGROUND: Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao is one of the most common herbs widely used in South and East Asia, to enhance people's health and reinforce vital energy. Despite its prevalence, however, the knowledge about phytochemical compositions and metabolite biosynthesis in Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao is very limited. RESULTS: An integrated metabolomics and transcriptomics analysis using state-of-the-art UPLC-Q-Orbitrap mass spectrometer and advanced bioinformatics pipeline were conducted to study global metabolic profiles and phytochemical ingredients/biosynthesis in Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. A total of 5435 metabolites were detected, from which 2190 were annotated, representing an order of magnitude increase over previously known. Metabolic profiling of Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao tissues found contents and synthetic enzymes for phytochemicals were significantly higher in leaf and stem in general, whereas the contents of the main bioactive ingredients were significantly enriched in root, underlying the value of root in herbal remedies. Using integrated metabolomics and transcriptomics data, we illustrated the complete pathways of phenylpropanoid biosynthesis, flavonoid biosynthesis, and isoflavonoid biosynthesis, in which some were first reported in the herb. More importantly, we discovered novel flavonoid derivatives using informatics method for neutral loss scan, in addition to inferring their likely synthesis pathways in Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. CONCLUSIONS: The current study represents the most comprehensive metabolomics and transcriptomics analysis on traditional herb Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. We demonstrated our integrated metabolomics and transcriptomics approach offers great potentials in discovering novel metabolite structure and associated synthesis pathways. This study provides novel insights into the phytochemical ingredients, metabolite biosynthesis, and complex metabolic network in herbs, highlighting the rich natural resource and nutritional value of traditional herbal plants.

MYB transcription factor PdMYB118 directly interacts with bHLH transcription factor PdTT8 to regulate wound-induced anthocyanin biosynthesis in poplar.

BACKGROUND: R2R3-MYB transcription factors (TFs) play important roles in plant growth and development, and response to biotic and abiotic stresses. However, their regulatory mechanisms in wound-induced anthocyanin biosynthesis in woody plants are largely unknown. RESULTS: In this work, we report that expression of anthocyanin biosynthesis genes (ABGs) were activated by PdMYB118, a MYB TF encoding gene from Populus deltoids, and the activation of PdMYB118 was significantly enhanced by PdTT8, a bHLH protein, through its direct interaction with PdMYB118. PdMYB118 and some ABGs were evidently induced by wound induction and methyl jasmonate (MeJA) treatment. Overexpression of PdMYB118 promoted anthocyanin accumulation in transgenic poplar upon wound induction. Furthermore, a poplar JASMONATE ZIM-domain (JAZ) protein, PtrJAZ1, repressed the transcriptional function of PdMYB118/PdTT8 complex by binding to PdTT8, and wound stimulated the biosynthesis of jasmonic acid (JA) and the degradation of PtrJAZ1. CONCLUSIONS: Based on these observations, we proposed that PtrJAZ1 degradation triggered the expression of ABGs, leading to increased biosynthesis of anthocyanins in the wounded leaves of transgenic poplar. Therefore, our findings not only illustrate the crucial role of PdMYB118 in wound-induced anthocyanin biosynthesis in poplar, but also provide a molecular basis for the genetic engineering of colorful tree species.

PdMYB118, isolated from a red leaf mutant of Populus deltoids, is a new transcription factor regulating anthocyanin biosynthesis in poplar.

KEY MESSAGE: A new anthocyanin biosynthesis transcription factor PdMYB118, which could be used for the genetic engineering of colorful tree species, was indentified from a red leaf mutant of Populus deltoids. In higher plants, the biosynthesis of anthocyanins is regulated by several classes of transcription factors (TFs), including R2R3-MYB, bHLH and WD-repeat proteins. In this work, we isolated an MYB gene regulating anthocyanin biosynthesis from a red leaf mutant of Populus deltoids, which accumulated more anthocyanins in the leaves and showed higher expression levels of anthocyanin biosynthesis genes than did the wild type. Gene expression analyses of all TFs regulating anthocyanin biosynthesis demonstrated that only a MYB118 homologous gene, PdMYB118, was up-regulated in the mutant compared with the wide type. Subcellular localization analyses in poplar leaf mesophyll protoplasts showed that PdMYB118-YFP fusion protein was specifically located in nucleus. When transiently expressed in poplar leaf protoplasts, PdMYB118 specifically promoted the expression of anthocyanidin biosynthesis genes. Dual-luciferase assays revealed that PdMYB118 can directly activate the promoters of these genes. When overexpressed in Shanxin Yang (P. davidiana × P. bolleana), a hybrid clone commercially grown for landscaping in the northern part of China, transgenic plants overexpressing PdMYB118 produced more anthocyanins in the leaves and turned their color into redness when grown in both greenhouse and field. Consistently, transcripts of some important anthocyanidin biosynthesis genes were significantly increased in the leaves of transgenic plants. All these results indicate that PdMYB118 functions as an essential transcription factor regulating anthocyanin biosynthesis in poplar and could be used for the genetic engineering of colorful tree species.

Ectopic expression of a cytochrome P450 monooxygenase gene PtCYP714A3 from Populus trichocarpa reduces shoot growth and improves tolerance to salt stress in transgenic rice.

In Arabidopsis thaliana and Oryza sativa, the cytochrome P450 (CYP) 714 protein family represents a unique group of CYP monooxygenase, which functions as a shoot-specific regulator in plant development through gibberellin deactivation. Here, we report the functional characterizations of PtCYP714A3, an OsCYP714D1/Eui homologue from Populus trichocarpa. PtCYP714A3 was ubiquitously expressed with the highest transcript level in cambium-phloem tissues, and was greatly induced by salt and osmotic stress in poplar. Subcellular localization analyses indicated that PtCYP714A3-YFP fusion protein was targeted to endoplasmic reticulum (ER). Expression of PtCYP714A3 in the rice eui mutant could rescue its excessive-shoot-growth phenotype. Ectopic expression of PtCYP714A3 in rice led to semi-dwarfed phenotype with promoted tillering and reduced seed size. Transgenic lines which showed significant expression of PtCYP714A3 also accumulated lower GA level than did the wild-type (WT) plants. The expression of some GA biosynthesis genes was significantly suppressed in these transgenic plants. Furthermore, transgenic rice plants exhibited enhanced tolerance to salt and maintained more Na(+) in both shoot and root tissues under salinity stress. All these results not only suggest a crucial role of PtCYP714A3 in shoot responses to salt toxicity in rice, but also provide a molecular basis for genetic engineering of salt-tolerant crops.

Antisense expression of the fasciclin-like arabinogalactan protein FLA6 gene in Populus inhibits expression of its homologous genes and alters stem biomechanics and cell wall composition in transgenic trees.

Fasciclin-like arabinogalactan proteins (FLAs) play important roles in the growth and development of roots, stems, and seeds in Arabidopsis. However, their biological functions in woody plants are largely unknown. In this work, we investigated the possible function of PtFLA6 in poplar. Quantitative real-time PCR, PtFLA6-yellow fluorescent protein (YFP) fusion protein subcellular localization, Western blotting, and immunohistochemical analyses demonstrated that the PtFLA6 gene was expressed specifically in the xylem of mature stem, and PtFLA6 protein was distributed ubiquitous in plant cells and accumulated predominantly in stem xylem fibres. Antisense expression of PtFLA6 in the aspen hybrid clone Poplar davidiana×Poplar bolleana reduced the transcripts of PtFLA6 and its homologous genes. Transgenic plants that showed a significant reduction in the transcripts of PtFLAs accumulated fewer PtFLA6 and arabinogalactan proteins than did the non-transgenic plants, leading to reduced stem flexural strength and stiffness. Further studies revealed that the altered stem biomechanics of transgenic plants could be attributed to the decreased cellulose and lignin composition in the xylem. In addition expression of some xylem-specific genes involved in cell wall biosynthesis was downregulated in these transgenic plants. All these results suggest that engineering the expression of PtFLA6 and its homologues could modulate stem mechanical properties by affecting cell wall composition in trees.

The tumor necrosis factor receptor-associated factor (TRAF)-like family protein SEVEN IN ABSENTIA 2 (SINA2) promotes drought tolerance in an ABA-dependent manner in Arabidopsis.

Tumor necrosis factor receptor-associated factor (TRAF) proteins play crucial roles in plant development and response to abiotic stress. Here, we present genetic evidence that SEVEN IN ABSENTIA 2 (SINA2), a TRAF-like family protein, is involved in abscisic acid (ABA)-related drought stress signaling in Arabidopsis. Gene expression, protein subcellular localization, protein-protein interaction, and a transient transcription dual-luciferase assay were performed. The drought tolerance of SINA2 loss-of-function mutants and SINA2-overexpressing plants was investigated. In Arabidopsis, SINA2 was significantly induced by ABA and drought treatment. The SINA2-YFP fusion protein was predominately localized in the nuclei and cytoplasm. Loss of function of SINA2 (sina2) reduced drought tolerance, whereas overexpression of SINA2 increased stomatal closure, decreased water loss, and therefore improved drought resistance in transgenic plants. Upon ABA treatment, expression of some key ABA- and stress-responsive genes decreased in the sina2 mutant, but increased in SINA2-overexpressing plants. Furthermore, SINA2 was induced in the ABA-deficient mutant by ABA, but not by drought stress. Thus, the drought response of SINA2 was ABA-dependent. ProSINA2::LUC expression in Arabidopsis protoplasts further revealed that ABA-responsive element (ABRE) binding (AREB) protein 1 (AREB1) AREB2 and ABRE-binding factor 3 (ABF3) might regulate SINA2 expression at the transcriptional level. Our results indicate that SINA2 functions as a positive molecular link between drought tolerance and ABA signaling in Arabidopsis.

Poplar calcineurin B-like proteins PtCBL10A and PtCBL10B regulate shoot salt tolerance through interaction with PtSOS2 in the vacuolar membrane.

The calcineurin B-like protein (CBL) family represents a unique group of calcium sensors in plants. In Arabidopsis, CBL10 functions as a shoot-specific regulator in salt tolerance. We have identified two CBL10 homologs, PtCBL10A and PtCBL10B, from the poplar (Populus trichocarpa) genome. While PtCBL10A was ubiquitously expressed at low levels, PtCBL10B was preferentially expressed in the green-aerial tissues of poplar. Both PtCBL10A and PtCBL10B were targeted to the tonoplast and expression of either one in the Arabidopsis cbl10 mutant could rescue its shoot salt-sensitive phenotype. Like PtSOS3, both PtCBL10s physically interacted with the salt-tolerance component PtSOS2. But in contrast to the SOS3-SOS2 complex at the plasma membrane, the PtCBL10-SOS2 interaction was primarily associated with vacuolar compartments. Furthermore, overexpression of either PtCBL10A or PtCBL10B conferred salt tolerance on transgenic poplar plants by maintaining ion homeostasis in shoot tissues under salinity stress. These results not only suggest a crucial role of PtCBL10s in shoot responses to salt toxicity in poplar, but also provide a molecular basis for genetic engineering of salt-tolerant tree species.

Metals Transfer in Mushroom Tricholoma matsutake from Regional High Geochemical Background Areas: Environmental Influences and Human Health Risk.

Wild-grown edible mushrooms are important in world diets and are also efficient metal accumulators. Yunnan, Southwest China, is the main producing region, with typically high levels of geochemical metals. The environmental factors, bioaccumulation, distribution and human health risks of metals were examined in paired soil and Tricholoma matsutake (n = 54). T. matsutake grows on acidified soils (pH = 3.95-6.56), and metals show a strong heterogeneity, with Fe, Mn, Zn and Cu in the ranges of 16-201, 0.046-8.58 g kg-1, and 22.6-215, 3.7-155 mg kg-1. High soil Fe content led to great accumulation in T. matsutake (0.24-18.8 g kg-1). However, though the soil Mn content was higher than that of Zn and Cu, their concentrations in T. matsutake were comparable (21.1-487 vs. 38.7-329 and 24.9-217 mg kg-1). This suggested that T. matsutake prefers to accumulate Zn and Cu compared to Mn, and this is supported by the bioaccumulation factors (BAFs = 0.32-17.1 vs. 0.006-1.69). Fe was mainly stored in stipes, while Mn, Zn and Cu were stored in caps, and the translocation factors (TFs) were 0.58 vs. 1.28-1.94. Therefore, stipe Fe showed the highest health risk index (HRI) at 1.28-26.9, followed by cap Cu (1.01-2.33), while 98-100% of the Mn and Zn were risk-free. The higher concentration and greater risk of Fe was attributed to the significant effect of soil Fe content (R = 0.34) and soil pH (R = -0.57). This study suggested that Fe, as an essential mineral, may exert toxic effects via the consumption of T. matsutake from high geochemical background areas.

Tangzhiping Decoction Improves Glucose and Lipid Metabolism and Exerts Protective Effects Against White Adipose Tissue Dysfunction in Prediabetic Mice.

Background: Prediabetes, characterized by a series of metabolic abnormalities, increases the risk of diabetes and cardiovascular diseases. Tangzhiping (TZP), a clinically validated traditional Chinese medicine formula, is used to treat impaired glucose tolerance. However, the underlying mechanism of TZP in intervening prediabetes is not fully elucidated. Purpose: The current study aimed to evaluate the protective effect of TZP against prediabetes mice and explore its potential mechanism. Methods: After establishing a prediabetic animal model through 12 weeks of high-fat diet (HFD) feeding, mice were subjected to TZP for 8 weeks. Various parameters related to body weight, glucose and lipid metabolism, and insulin sensitivity were measured. Histopathological examinations observed adipose cell size and liver lipid deposition. The Sable Promethion system assessed energy metabolism activity. Transcriptomic analysis of Epididymal white adipose tissue (EWAT) identified enriched pathways and genes. The key genes in the enriched pathways were identified through RT-PCR. Results: Our data revealed that the administration of TZP reduced body weight and fat mass in a prediabetes mouse model. TZP normalized the glucose and insulin levels, improved insulin resistance, and decreased plasma TC and FFA. The alleviation of adipose tissue hypertrophy and lipid deposition by TZP was demonstrated through pathological examination. Indirect calorimetry measurements indicated a potential increase in VO2 and EE levels with TZP. The results of EWAT transcription showed that TZP reversed pathways and genes related to inflammation and catabolic metabolism. RT-PCR demonstrated that the mRNA expression of inflammation and lipolysis, including Tlr2, Ccr5, Ccl9, Itgb2, Lipe, Pnpla2, Cdo1, Ces1d, Echs1, and Acad11, were changed by TZP treatment. Conclusion: TZP effectively alleviates obesity, impaired glucose and lipid metabolism, and insulin resistance. The effect of TZP might be associated with the regulation of gene expression in dysfunctional adipose tissue.

Introduction of the rice CYP714D1 gene into Populus inhibits expression of its homologous genes and promotes growth, biomass production and xylem fibre length in transgenic trees.

The rice (Oryza sativa) OsCYP714D1 gene (also known as EUI) encodes a cytochrome P450 monooxygenase which functions as a gibberellin (GA)-deactivating enzyme, catalysing 16α, 17-epoxidation of non-13-hydroxylated GAs. To understand whether it would also reduce the production of active GAs and depress the growth rate in transgenic trees, we constitutively expressed OsCYP714D1 in the aspen hybrid clone Populus alba×P. berolinensis. Unexpectedly, ectopic expression of OsCYP714D1 in aspen positively regulated the biosynthesis of GAs, including the active GA1 and GA4, leading to promotion of the growth rate and biomass production in transgenic plants. Transgenic lines which showed significant expression of the introduced OsCYP714D1 gene accumulated a higher GA level and produced more numerous and longer xylem fibres than did the wild-type plants. Quantitative real-time PCR indicated that transcription of most homologous PtCYP714 genes was suppressed in these transgenic lines. Therefore, the promoted GA and biomass production in transgenic trees constitutively expressing OsCYP714D1 is probably attributed to the down-regulated expression of the native PtCYP714 homologues involved in the GA biosynthesis pathway, although their precise functions are yet to be further elucidated.

Choledochoscope-guided treatment of pulmonary embolism caused by ventricular myxoma.

A 33-year male patient presented with a 6-month history of cough and shortness of breath upon physical activity. Echocardiography demonstrated right ventricular space-occupying lesions. Contrast-enhanced computed tomography of the chest showed multiple emboli in the pulmonary artery and its branches. Right ventricle tumor (myxoma) resection, tricuspid valve replacement, and clearance of the pulmonary artery thrombus were performed under cardiopulmonary bypass. Minimally invasive forceps and balloon urinary catheters were used to clear the thrombus. Clearance was confirmed by direct visualization using a choledochoscope. The patient recovered well and was discharged. The patient was prescribed oral warfarin 3 mg/day, and the international normalized ratio for prothrombin time was maintained between 2.0 and 3.0. Pre-discharge echocardiogram showed no lesion in the right ventricle or pulmonary arteries. The 6-month follow-up echocardiography indicated that the tricuspid valve was functioning well and showed no thrombus in the pulmonary artery.

Expectation-maximization algorithm leads to domain adaptation for a perineural invasion and nerve extraction task in whole slide digital pathology images.

In addition to lymphatic and vascular channels, tumor cells can also spread via nerves, i.e., perineural invasion (PNI). PNI serves as an independent prognostic indicator in many malignancies. As a result, identifying and determining the extent of PNI is an important yet extremely tedious task in surgical pathology. In this work, we present a computational approach to extract nerves and PNI from whole slide histopathology images. We make manual annotations on selected prostate cancer slides once but then apply the trained model for nerve segmentation to both prostate cancer slides and head and neck cancer slides. For the purpose of multi-domain learning/prediction and investigation on the generalization capability of deep neural network, an expectation-maximization (EM)-based domain adaptation approach is proposed to improve the segmentation performance, in particular for the head and neck cancer slides. Experiments are conducted to demonstrate the segmentation performances. The average Dice coefficient for prostate cancer slides is 0.82 and 0.79 for head and neck cancer slides. Comparisons are then made for segmentations with and without the proposed EM-based domain adaptation on prostate cancer and head and neck cancer whole slide histopathology images from The Cancer Genome Atlas (TCGA) database and significant improvements are observed.

Expanding the Coverage of Metabolic Landscape in Cultivated Rice with Integrated Computational Approaches.

Genome-scale metabolomics analysis is increasingly used for pathway and function discovery in the post-genomics era. The great potential offered by developed mass spectrometry (MS)-based technologies has been hindered, since only a small portion of detected metabolites were identifiable so far. To address the critical issue of low identification coverage in metabolomics, we adopted a deep metabolomics analysis strategy by integrating advanced algorithms and expanded reference databases. The experimental reference spectra and in silico reference spectra were adopted to facilitate the structural annotation. To further characterize the structure of metabolites, two approaches were incorporated into our strategy, i.e., structural motif search combined with neutral loss scanning and metabolite association network. Untargeted metabolomics analysis was performed on 150 rice cultivars using ultra-performance liquid chromatography coupled with quadrupole-Orbitrap MS. Consequently, a total of 1939 out of 4491 metabolite features in the MS/MS spectral tag (MS2T) library were annotated, representing an extension of annotation coverage by an order of magnitude in rice. The differential accumulation patterns of flavonoids between indica and japonica cultivars were revealed, especially O-sulfated flavonoids. A series of closely-related flavonolignans were characterized, adding further evidence for the crucial role of tricin-oligolignols in lignification. Our study provides an important protocol for exploring phytochemical diversity in other plant species.

An efficient Agrobacterium-mediated transformation and regeneration system for leaf explants of two elite aspen hybrid clones Populus alba × P. berolinensis and Populus davidiana × P. bolleana.

Transgenic technology has been successfully used for gene function analyses and trait improvement in cereal plants. However, its usage is limited in woody plants, especially in the difficult-to-transform but commercially viable hybrid poplar. In this work, an efficient regeneration and transformation system was established for the production of two hybrid aspen clones: Populus alba × P. berolinensis and Populus davidiana × P. bolleana. A plant transformation vector designed to express the reporter gene uidA, encoding ß-glucuronidase (GUS), driven by the cauliflower mosaic virus 35S promoter, was used to detect transformation event at early stages of plant regeneration, and to optimize the parameters that may affect poplar transformation efficiency. Bacterium strain and age of leaf explant are two major factors that affect transformation efficiency. Addition of thidiazuron (TDZ) improved both regeneration and transformation efficiency. The transformation efficiency is approximately 9.3% for P. alba × P. berolinensis and 16.4% for P. davidiana × P. bolleana. Using this system, transgenic plants were usually produced in less than 1 month after co-cultivation. The growth characteristics and morphology of transgenic plants were identical to the untransformed wild type plants, and the transgenes could be inherited by vegetative propagation, as confirmed by PCR, Southern blotting, RT-PCR and ß-glucuronidase staining analyses. The establishment of this system will help to facilitate the studies of gene functions in tree growth and development at a genome level, and as well as the introduction of some valuable traits in aspen breeding.

Toxic effects of 2,4,4'- trichlorobiphenyl (PCB-28) on growth, photosynthesis characteristics and antioxidant defense system of Lemna minor L.

Polychlorinated biphenyls (PCBs) are a common category of persistent man-made organic pollutants that are widespread in the ambient environment. Although Lemna minor L. is an extensively applied plant for aquatic remediation in ecotoxicology research worldwide, little is known regarding its responses to the potentially toxic effects of PCBs. For this study, a 14-day dissolved exposure was conducted to explore the effects of 2,4,4'- trichlorobiphenyl (PCB-28) on the growth, photosynthesis characteristics and antioxidant defense system of L. minor plants. We found that 100 and 200 µg/L of PCB-28 decreased the fresh weight, chlorophyll and protein content, and activities of superoxide dismutase, peroxidase, glutathione S-transferase, and nitroreductase, whereas plasma membrane permeability, and the malondialdehyde and reactive oxygen species concentrations were increased. However, it was observed that 5 and 20 µg/L of PCB-28 had no significant effects on these physiological indices. The ultra-structure of chloroplast demonstrated that 100 and 200 µg/L PCB-28 severely damaged the chloroplast structures. Moreover, correlation analysis revealed that the content of reactive oxygen species had negative correlations with the fresh weight, chlorophyll and protein content, as well as the activities of superoxide dismutase, peroxidase, glutathione S-transferase, and nitroreductase, but had positive correlations with the malondialdehyde content and plasma membrane permeability. This work provides valuable data toward elucidating the physiology and biochemistry of PCBs induced phytotoxicity.

Daidzein Alleviates Hypothalamic-Pituitary-Adrenal Axis Hyperactivity, Ameliorates Depression-Like Behavior, and Partly Rectifies Circulating Cytokine Imbalance in Two Rodent Models of Depression.

Depression is one very common mental health disorder which can cause morbidity and mortality if not addressed. Recent studies have provided strong evidence that depression may be accompanied by immune activation, secondary inflammatory reaction, and hyperactivity of the Hypothalamic Pituitary Adrenal (HPA) axis. It is well-known that it takes at least 2 weeks for conventional antidepressants, especially SSRIs (Selective serotonin reuptake inhibitors) to produce effects. To better understand the mechanism of antidepressant effects on depression and subsequently further elucidate the pathogenesis of depression, we selected phytestrogen daidzein (DD) to observe its effects on the depression-like and anxiety-like behavior in two different rodent models of depression which were induced by learned helplessness and chronic mild stress (CMS) and then simultaneous evaluation of the depression-like behavior, the activity of HPA axis, and circulatory cytokines. Our results showed that daidzein attenuated depression-like behaviors through alleviating HPA axis hyperactivity, decreasing the levels of stress-related hormones, and partly rectifying some inflammatory cytokines imbalance in both the rodent models of depression.

Spinal Tuina Improves Cognitive Impairment in Cerebral Palsy Rats through Inhibiting Pyroptosis Induced by NLRP3 and Caspase-1.

Cerebral palsy (CP) is a severe cerebral disease with high mortality and morbidity, which leads to great challenges for the suffering children and their families. Hence, the need for the efficacious and safe treatments is urgent. As a physical therapy arising from traditional Chinese medicine (TCM), Tuina has shown multiple effects on various diseases, including cerebral palsy. Nevertheless, the detailed mechanisms of Tuina on CP remain unknown, which impedes its further clinical application. Herein, we explored the effects of Tuina on CP and its potential mechanisms. Thirty Sprague Dawley (SD) male rats were randomly divided into sham, model, and Tuina groups (model + Tuina). CP rat model was established by hypoxia-ischemia via permanent occlusion of left common carotid artery and hypoxia for 2.5 hours caused by anaerobic environment, which was subsequently followed by onset of Tuina treatment from postnatal day 7 (P7) to P49. After completion of Tuina treatment, the behavioral tests showed that Tuina treatment not only improved the retarded body weight and impaired motor balance function, but also ameliorated weakened learning and memory function of CP rats. Moreover, immunohistochemistry and western blot also revealed a reduced expression of NLRP3 inflammasome and corresponding pyroptosis-related molecules induced by NLRP3 in CP rats after Tuina treatment. Therefore, our study indicated that Tuina treatment may improve impaired neurocognitive function of CP rats, which was possibly realised via inhibiting NLRP3-induced pyroptosis.