Asymmetric response of aboveground and belowground temporal stability to nitrogen and phosphorus addition in a Tibetan alpine grassland.

Anthropogenic eutrophication is known to impair the stability of aboveground net primary productivity (ANPP), but its effects on the stability of belowground (BNPP) and total (TNPP) net primary productivity remain poorly understood. Based on a nitrogen and phosphorus addition experiment in a Tibetan alpine grassland, we show that nitrogen addition had little impact on the temporal stability of ANPP, BNPP, and TNPP, whereas phosphorus addition reduced the temporal stability of BNPP and TNPP, but not ANPP. Significant interactive effects of nitrogen and phosphorus addition were observed on the stability of ANPP because of the opposite phosphorus effects under ambient and enriched nitrogen conditions. We found that the stability of TNPP was primarily driven by that of BNPP rather than that of ANPP. The responses of BNPP stability cannot be predicted by those of ANPP stability, as the variations in responses of ANPP and BNPP to enriched nutrient, with ANPP increased while BNPP remained unaffected, resulted in asymmetric responses in their stability. The dynamics of grasses, the most abundant plant functional group, instead of community species diversity, largely contributed to the ANPP stability. Under the enriched nutrient condition, the synchronization of grasses reduced the grass stability, while the latter had a significant but weak negative impact on the BNPP stability. These findings challenge the prevalent view that species diversity regulates the responses of ecosystem stability to nutrient enrichment. Our findings also suggest that the ecological consequences of nutrient enrichment on ecosystem stability cannot be accurately predicted from the responses of aboveground components and highlight the need for a better understanding of the belowground ecosystem dynamics.

The role of post-transcriptional modification on a new tRNAIle(GAU) identified from Ganoderma lucidum in its fragments' cytotoxicity on cancer cells.

Ganoderma lucidum (Ganoderma) is a famous Chinese herbal medicine which has been used clinically for thousands of years in China. Despite numerous studies on triterpenes and polysaccharides, the bioactivity of RNAs abundant in Ganoderma remains unknown. Here, based on LC-MS techniques, dihydrouracil, 5-methyluridine (m5U) and pseudouridine were identified at position 19, 52 and 53 of a new tRNAIle(GAU) which was isolated as the most abundant tRNA species in Ganoderma, and is the first purified tRNA from fungus. Cytotoxic screening of tRNA-half (t-half) and tRNA fragment (tRF) derived from this tRNA, as well as their mimics (t-half or tRF as antisense strand), demonstrated that the double-stranded form, i.e., tRF and t-halve mimics, exhibited stronger cytotoxicity than their single-stranded form, and the cytotoxicity of t-half mimic is significantly stronger than that of tRF mimic. Notably, the cytotoxicity of 3'-t-half mimic is not only much more potent than that of taxol, but also is much more potent than that of ganoderic acids, the major bioactive components in Ganoderma. Furthermore, 3'-t-half mimic_M2 (m5U modified) exhibited significantly stronger cytotoxicity than unmodified 3'-t-half mimic, which is consistent with the computational simulation showing that m5U modification enhances the stability of the tertiary structure of 3'-t-half mimic. Overall, the present study not only indicates t-halves are bioactive components in Ganoderma which should not be neglected, but also reveals an important role of post-transcriptional modification on tRNA in its fragments' cytotoxicity against cancer cells, which benefits the design and development of RNAi drugs from natural resource.

The prognostic value of IPI in patients with primary breast lymphoma, a multicenter retrospective study.

BACKGROUND: Due to the rarity of PBL and the lack of large-scale studies, the prognostic value of IPI in PBL was controversial. Especially in the rituximab era, the ability of IPI to stratify prognosis in patients receiving immunochemotherapy was severely reduced. Then revised IPI (R-IPI) and National Comprehensive Cancer Network IPI (NCCN-IPI) were introduced. The present study aimed to evaluate the prognostic value of IPI and the other IPIs in patients with PBL in a Chinese population. METHODS: We performed a multicenter retrospective study of 71 patients with PBL from 3 institutions in China. The Kaplan-Meier method and log-rank tests were used for the survival analysis. Cox regression analysis was performed to evaluate the prognostic factors. Subgroup analysis was performed to assess the prognostic significance of IPI scores, R-IPI scores, and NCCN-IPI scores. RESULTS: The median follow-up was 4.7 years (0.7-21.8 years). The 5-year progression-free survival (PFS) and overall survival (OS) rates were 90.2% and 96.3%. In the multivariate analysis, only IPI scores and radiotherapy were significantly associated with OS and PFS (P < 0.05). Applying the R-IPI in our patient cohort indicates a significant difference in PFS between the two groups of R-IPI (P = 0.034) but not for OS (P = 0.072). And the NCCN-IPI was prognostic for OS (P = 0.025) but not for PFS (P = 0.066). Subgroup analyses of IPI showed that survival analysis of IPI scores for the PFS and OS of patients using rituximab were not significantly different (P > 0.05). CONCLUSIONS: Our study confirms the prognostic value of IPI in patients with PBL, but the predictive value of IPI proved to be relatively low with the addition of the rituximab. The R-IPI and NCCN-IPI can accurately assess the high and low-risk groups of PBL patients but were insufficient to evaluate the intermediate risk group.

The Effects of Tai Chi on the Executive Functions and Physical Fitness in Middle-Aged Adults with Depression: A Randomized Controlled Trial.

Objective: The present study examined the effects of Tai Chi exercise on the executive functions (EFs) and physical fitness of middle-aged adults with depression. Methods: A total of 39 middle-aged adults with depression (M age = 50.59, SD = 7.38) were randomly assigned to the Tai Chi group (n = 20) or the waiting-list control group (n = 19). The Tai Chi group engaged in two 90 min sessions of Tai Chi exercise per week for 12 weeks; the waiting-list control group was asked to maintain their usual daily routines for 12 weeks. Depression symptoms, EFs (i.e., inhibitory control, planning, working memory, and cognitive flexibility), and physical fitness (i.e., cardiovascular fitness, muscular strength, muscular endurance, power, and flexibility) were evaluated at the baseline (pretest), 6-week (mid-test), and 12-week (post-test) marks. Results: Both groups showed decreased depression symptoms over time. Compared with the control group, the Tai Chi group showed decreased reaction times for incongruent conditions in the Stroop test from pretest to mid- and post-test, and shorter reaction time for incongruent conditions in the Stroop test than the control group at post-test; the Tai Chi group performed significantly better than the control group in overall total move score of Tower of London (TOL). The Tai Chi group also showed increased total correct scores of TOL from pretest to mid- and post-test, and greater total correct scores of TOL than the control group at post-test. Additionally, results indicated that Tai Chi exercise comprehensively improved physical fitness from pretest to mid- and post-test. Greater performance in terms of cardiovascular fitness, muscular strength, and power was also found in the Tai Chi group at post-test than in the control group. Conclusions: These findings suggest that the 12-week Tai Chi exercise improved inhibitory control, planning and working memory aspects of executive functions, and physical fitness in middle-aged adults with depression.

[Effect of moxibustion on autophagy lysosome function mediated by mTOR/TFEB pathway and lncRNA H19 expression in APP/PS1 double transgenic mice].

OBJECTIVE: To observe the effect of moxibustion (Moxi) at acupoints of Governor Vessel on autophagy lysosomal function and lncRNA H19 in amyloid precursor protein/presenilin 1 (APP/PS1) double transgenic Alzheimer's disease (AD) mice, so as to explore its underlying mechanisms in relieving AD. METHODS: Fifty two male APP/PS1 double transgenic AD mice were randomly divided into model, Moxi, Moxi+inhibitor and medication (rapamycin) groups, with 13 mice in each group. Other 13 male C57BL/6J mice of the same age were selected as the control group. The mice of the Moxi group received aconite cake-separated Moxi stimulation at "Baihui" (GV20), "Dazhui"(GV14) and "Fengfu" (GV16), for 15 min, those of the Moxi+inhibitor group received intraperitoneal injection of 3-methyladenine (an inhibitor of PI3K for suppressing autophagy) 1.5 mg· kg-1 · d-1 on the basis of Moxi, and those of the medication group received intraperitoneal injection of rapamycin 2 mg· kg-1 · d-1. The treatment was conducted once daily for 2 weeks. The mouse's learning-memory ability was detected by Morris water maze tests. The hippocampus tissue was sampled for observing the formation of autophagy by using transmission electron microscope, detecting the expression of Aß_(1-42) protein with immunohistochemical staining, and for detecting the expression levels of long noncoding RNA H19 (lncRNA H19), mammalian target of rapamycin kinase (mTOR), nuclear transcription factor EB (TFEB), Cathepsin D and lysosome associated membrane protein-1 (LAMP1) genes and proteins as well as microtubule associated protein 1 light chain 3B (LC3B)-Ⅱ/LC3B-Ⅰand autophagy protein p62 protein by quantitative real-time PCR and Western blot, respectively. RESULTS: In contrast to the control group, the model group had an evident increase in the escape latency of Morris water maze test, and in the expression levels of Aß_(1-42) protein, lncRNA H19 mRNA, mTOR mRNA and protein, and p62 protein (P<0.05), and a significant decrease in the expression levels of TFEB, Cathepsin D, LAMP1 mRNAs and proteins and LC3B-Ⅱ/LC3B-Ⅰ (P<0.05). After the treatment and relevant to the model and Moxi+inhibitor groups, both the Moxi and medication groups had an obvious down-regulation in the levels of latency of Morris water maze, expression levels of Aß_(1-42) protein, lncRNA H19 mRNA, mTOR mRNA and protein, and p62 protein (P<0.05), and a significant up-regulation in the levels of TFEB, Cathepsin D, LAMP1 mRNAs and proteins and LC3B-Ⅱ/LC3B-Ⅰ (P<0.05). CONCLUSION: Moxi at acupoints of Governor Vessel can improve cognitive function of AD mice, which may be associated with its functions in inhibiting mTOR/TFEB pathway by down-regulating the expression of lncRNA H19, improving autophagy lysosomal function, promoting autophagy and clearing away Aß1-42 in the hippocampus.

Autophagy Regulates Whitefly-Symbiont Metabolic Interactions.

Nutritional symbionts are restricted to specialized host cells called bacteriocytes in various insect orders. These symbionts can provide essential nutrients to the host. However, the cellular mechanisms underlying the regulation of these insect-symbiont metabolic associations remain largely unclear. The whitefly Bemisia tabaci MEAM1 hosts "Candidatus Portiera aleyrodidarum" (here, "Ca. Portiera") and "Candidatus Hamiltonella defensa" (here, "Ca. Hamiltonella") bacteria in the same bacteriocyte. In this study, the induction of autophagy by chemical treatment and gene silencing decreased symbiont titers and essential amino acid (EAA) and B vitamin contents. In contrast, the repression of autophagy in bacteriocytes via Atg8 silencing increased symbiont titers, and amino acid and B vitamin contents. Furthermore, dietary supplementation with non-EAAs or B vitamins alleviated autophagy in whitefly bacteriocytes, elevated TOR (target of rapamycin) expression, and increased symbiont titers. TOR silencing restored symbiont titers in whiteflies after dietary supplementation with B vitamins. These data suggest that "Ca. Portiera" and "Ca. Hamiltonella" evade autophagy of the whitefly bacteriocytes by activating the TOR pathway via providing essential nutrients. Taken together, we demonstrate that autophagy plays a critical role in regulating the metabolic interactions between the whitefly and two intracellular symbionts. Therefore, this study reveals that autophagy is an important cellular basis for bacteriocyte evolution and symbiosis persistence in whiteflies. The whitefly symbiosis unravels the interactions between cellular and metabolic functions of bacteriocytes. IMPORTANCE Nutritional symbionts, which are restricted to specialized host cells called bacteriocytes, can provide essential nutrients for many hosts. However, the cellular mechanisms of regulation of animal-symbiont metabolic associations have been largely unexplored. Here, using the whitefly-"Ca. Portiera"/"Ca. Hamiltonella" endosymbiosis, we demonstrate autophagy regulates the symbiont titers and thereby alters the essential amino acid and B vitamin contents. For persistence in the whitefly bacteriocytes, "Ca. Portiera" and "Ca. Hamiltonella" alleviate autophagy by activating the TOR (target of rapamycin) pathway through providing essential nutrients. Therefore, we demonstrate that autophagy plays a critical role in regulating the metabolic interactions between the whitefly and two intracellular symbionts. This study also provides insight into the cellular basis of bacteriocyte evolution and symbiosis persistence in the whitefly. The mechanisms underlying the role of autophagy in whitefly symbiosis could be widespread in many insect nutritional symbioses. These findings provide a new avenue for whitefly control via regulating autophagy in the future.

Hierarchical dual-responsive cleavable nanosystem for synergetic photodynamic/photothermal therapy against melanoma.

Currently, the combining photodynamic therapy (PDT) with photothermal therapy (PTT) modalities based on a single near infrared (NIR) laser irradiation and highly selective internalization still remain a challenge. Herein, a hierarchical dual-responsive cleavable nanosystem for synergetic NIR triggered PDT/PTT is reported. The engineered nanoplatform (Au NRs/Cur/UCNPs@PBE) is designed by loading curcumin (Cur, photosensitizer) on gold nanarods (Au NRs) to build PDT/PTT therapy system, which was encapsulated outside with upconversion nanoparticles (UCNPs) and then modified with phenylboronic double ester (PBE). The pH and ROS-responsive feature made Au NRs/Cur/UCNPs@PBE provide a fundamental structural evolution and improve the specificity and intracellular accumulation to tumors. Au NRs/Cur/UCNPs@PBE exhibited significant PDT and PTT efficiency against two type melanoma cells due to upconversion nanoparticles and Au NRs induced by an 808 nm laser. Notably, the platform can mainly activate apoptosis and partial ferroptosis to achieve the synergistic PDT/PTT, furthermore, the integrated PDT with PTT using Au NRs/Cur/UCNPs@PBE showcased a great antitumor efficacy in vivo superior to the other alone treatment. Our findings highlight that this intelligent nanoagents for synergistic phototherapy facilitate enhanced fighting melanoma and provide a promising strategy for melanoma theranostics.

Traditional Chinese medicine for septic patients undergoing ulinastatin therapy: A meta-analysis.

PURPOSE: This study aimed to assess the efficacy of traditional Chinese medicine (TCM) in septic patients treated with ulinastatin. METHODS: PubMed, EmBase, and the Cochrane library were searched up to January 2021 to identify randomized controlled trials. The weight mean difference (WMD) and relative risk (RR) with 95% confidence intervals were used with the random-effects model. RESULTS: Twenty-three randomized controlled trials with 1903 septic patients were included. TCM significantly reduced the APACHE II score (WMD: -5.18; P < .001), interleukin-6 (WMD: -63.00; P < .001), tumor necrosis factor-α (WMD: -8.86; P < .001), c-reactive protein (WMD: -9.47; P < .001), mechanical ventilation duration (WMD: -3.98; P < .001), intensive care unit stay (WMD: -4.18; P < .001), procalcitonin (WMD: -0.53; P < .001), lipopolysaccharide (WMD: -9.69; P < .001), B-type natriuretic peptide (WMD: -159.87; P < .001), creatine kinase isoenzyme MB (WMD: -45.67; P < .001), cardiac troponin I (WMD: -0.66; P < .001), and all-cause mortality risk (RR: 0.55; P < .001). CONCLUSIONS: TCM lowers inflammation levels and reduces the risk of all-cause mortality for septic patients.

Relationship between Mindfulness, Psychological Skills, and Mental Toughness in College Athletes.

Numerous studies have shown that dispositional mindfulness is positively associated with many mental abilities related to sports performance, including psychological skills and mental toughness. The purpose of this study was to explore the relationship between dispositional mindfulness, psychological skills, and mental toughness among different types of athletes. For this cross-sectional study, 101 college athletes were recruited. Their dispositional mindfulness, psychological skills, and mental toughness were measured by the Mindfulness Attention Awareness Scale (MAAS), Athletic Psychological Skills Inventory (APSI), and Traits of Mental Toughness Inventory for Sports Scale (TMTIS). Pearson's correlation was used to calculate how dispositional mindfulness is associated with psychological skills and mental toughness. The results revealed that dispositional mindfulness is positively associated with comprehensive APSI (r = 0.21-0.36, p < 0.05), TMTIS overall (r = 0.27, p < 0.01), positive effort (r = 0.26, p = 0.01), and pressure (r = 0.30, p < 0.01). These findings suggest a positive linkage between mindfulness and the two examined psychological characteristics related to sports performance. Other approaches to increase mindfulness may be considered in the future.

Effects of Chinese Mind-Body Exercises on Executive Function in Middle-Aged and Older Adults: A Systematic Review and Meta-Analysis.

Chinese mind-body exercises (CMBEs) are positively associated with executive function (EF), but their effects on EF, from synthesized evidence using systematic and meta-analytic reviews, have not been conducted. Therefore, the present systematic review with meta-analysis attempted to determine whether CMBEs affect EF and its sub-domains, as well as how exercise, sample, and study characteristics moderate the causal relationship between CMBEs and EF in middle-aged and older adults. Seven electronic databases were searched for relevant studies published from the inception of each database through June 2020 (PubMed, Web of Science, Embase, Cochrane Controlled Trials Register, Wanfang, China National Knowledge Infrastructure, and Weipu). Randomized controlled trials with at least one outcome measure of CMBEs on EF in adults of mean age ≥ 50 years with intact cognition or mild cognitive impairment (MCI) and with or without chronic diseases were included. A total of 29 studies (N = 2,934) ultimately were included in this study. The results indicated that CMBEs improved overall EF (Standardized Mean Differences = 0.28, 95% CI 0.12, 0.44), as well as its sub-domains of working memory and shifting. The beneficial effects of CMBEs on EF occurred regardless of type (Tai Chi, Qigong), frequency of group classes (≤2 time, 3-4 time, ≥5 times), session time (≤45 min, 46-60 min), total training time (≥150 to ≤300 min, >300 min), and length of the CMBEs (4-12 week, 13-26 week, and >26 week), in addition to that more frequent participation in both group classes and home practice sessions (≥5 times per week) resulted in more beneficial effects. The positive effects of CMBEs on EF were also demonstrated, regardless of participants mean age (50-65 years old, >65 years old), sex (only female, both), and cognitive statuses (normal, MCI, not mentioned), health status (with chronic disease, without chronic disease), as well as training mode (group class, group class plus home practice) and study language (English, Chinese). This review thus suggests that CMBEs can be used as an effective method with small to moderate and positive effects in enhancing EF, and that more frequent group classes and home practice sessions may increase these effects. However, certain limitations, including strictly design studies, limited ES (effect size) samples for specific variables, and possible biased publications, required paying particular attention to, for further exploring the effects of CMBEs on EF.

Effect of annealing and heat-moisture pretreatments on the oil absorption of normal maize starch during frying.

The impacts of two hydrothermal pretreatments, annealing (ANN) and heat moisture treatment (HMT), on oil-absorption by normal maize starch (NMS) during frying were investigated using low-field nuclear magnetic resonance (LF-NMR). The structural organizations of the fried samples were also evaluated using SEM, XRD, ATR-FTIR, and DSC, respectively. Both hydrothermal pretreatments significantly reduced the total oil content in the starch after frying, with the magnitude of the effect depending on the treatment conditions used. SEM showed that the pretreated fried starch granules preserved more of their original morphology. XRD, FTIR, and DSC showed that both pretreatments preserved more of the short-range double helices and long-range organizations within the orthorhombic crystalline structure for NMS during frying. The promoting effect of ANN/HMT on the interactions of starch molecules and the rearrangement of double helices were hypothesized to be responsible for the increased thermal stability of starch granules in the present work. As a result, fried starch pretreated by ANN/HMT were more organized and more compact than fried NMS, thus inhibiting oil absorption during frying.

UPLC-ESI-MS/MS Analysis and Evaluation of Antioxidant Activity of Total Flavonoid Extract from Paeonia lactiflora Seed Peel and Optimization by Response Surface Methodology (RSM).

In this study, the ultrasound-assisted extraction (UAE) of flavonoid from Paeonia lactiflora seed peel was optimized by response surface methodology (RSM). Single-factor experiments and a three-factor three-level Box-Behnken design (BBD) were performed to explore the effects of the following parameters on flavonoid extraction: ethanol concentration (X 1), liquid-solid ratio (X 2), and ultrasonic time (X 3). The results showed that the optimal flavonoid yield (10.9045 mg RE/g) was as follows: ethanol concentration 62.93%, ultrasonic time 64.56 min, and liquid-solid ratio 24.86 mL/g. The optimized extract of P. lactiflora seed shell was further analyzed by UPLC-ESI-MS/MS, and 20 main flavonoids were identified and quantified, among which protocatechuic acid, vanillic acid, 4-hydroxybenzoic acid, and 3,4-dihydroxybenzaldehyde had the highest content. Furthermore, the results of the antioxidant test showed that the P. lactiflora seed peel extract obtained under optimized UAE conditions exhibited good antioxidant activity. The experimental results showed that ultrasound-assisted extraction was a fast, efficient, and simple method for extracting active ingredients from P. lactiflora seed peel, thereby making this byproduct a promising source of compounds in food and healthcare sectors.

Grape seed proanthocyanidins improves depression-like behavior by alleviating oxidative stress and NLRP3 activation in the hippocampus of prenatally-stressed female offspring rats.

Over several decades, there is a growing evidence, which has shown that prenatal stress (PS) contributes to depression in offspring. Grape seed proanthocyanidins (GSPs), which contain dimers, trimers, oligomers of catechin and epicatechin, are known to possess antidepressant effects. The present study aimed to investigate the mechanism of antidepressant effects of GSPs on female juvenile prenatally stressed offspring rats. The results showed that the female juvenile offspring rats exposed to PS exhibited depression-like behavior manifested as longer immobility time and lesser consumption of sucrose solution. Prenatal stress reduced the number of hippocampal neurons and increased the level of the reactive oxygen species (ROS) in the hippocampus of the female juvenile offspring rats. Furthermore, the expression of PYD domains-containing protein 3 (NLRP3) and its downstream cytokines, Caspase-1, and interleukin-1ß (IL-1ß), were increased in the hippocampus of the female juvenile offspring rats exposed to PS. Administration of GSPs not only improved depression-like behavior and enhanced the number of hippocampal neurons, but also abated excessive ROS generation and inhibited the activation of the NLRP3-Caspase-1 signaling pathway. Taken together, GSPs counteract PS-induced hippocampal neuron loss and depression-like behavior by alleviating oxidative stress and NLRP3 activation. The present study provides a new insight for GSPs as an effective therapeutic agent for adolescent depression.

Pantothenate mediates the coordination of whitefly and symbiont fitness.

Intracellular symbionts in insects often have reduced genomes. Host acquisition of genes from bacteria is an important adaptation that supports symbionts. However, the function of horizontally transferred genes in insect symbiosis remains largely unclear. The primary symbiont Portiera housed in bacteriocytes lacks pantothenate synthesis genes: panB and panC, which is presumably complemented by a fused gene panB-panC (hereafter panBC) horizontally transferred from bacteria in Bemisia tabaci MEAM1. We found panBC in many laboratory cultures, and species of B. tabaci shares a common evolutionary origin. We demonstrated that complementation with whitefly panBC rescued E. coli pantothenate gene knockout mutants. Portiera elimination decreased the pantothenate level and PanBC abundance in bacteriocytes, and reduced whitefly survival and fecundity. Silencing PanBC decreased the Portiera titer, reduced the pantothenate level, and decreased whitefly survival and fecundity. Supplementation with pantothenate restored the symbiont titer, PanBC level, and fitness of RNAi whiteflies. These data suggest that pantothenate synthesis requires cooperation and coordination of whitefly PanBC expression and Portiera. This host-symbiont co-regulation was mediated by the pantothenate level. Our findings demonstrated that pantothenate production, by the cooperation of a horizontally acquired, fused bacteria gene and Portiera, facilitates the coordination of whitefly and symbiont fitness. Thus, this study extends our understanding on the basis of complex host-symbiont interactions.

Analysis of porous structure of potato starch granules by low-field NMR cryoporometry and AFM.

Pore size distribution is a crucial structural element affecting the adsorption and diffusion of reagents and enzymes within starch granules. An accurate and credible method of determining the pore size distribution of starch granules especially for smooth ones is therefore required. In this work, low-field NMR cryoporometry (LF-NMRC) was applied to analyze the pore structure of potato starch (PS). The reliability of the LF-NMRC method is verified by comparing with the traditional method, i.e. the low temperature nitrogen adsorption (LT-NA). Both LF-NMRC and LT-NA could characterize the PS pore structure in mesoporous range. However, LF-NMRC has superiority over LT-NA in terms of the distinguishment and determination of pore size distribution approaching to the micropores, gives more accurate and reliable results than LT-NA does. Structural evidences from scanning electron microscope (SEM) and atomic force microscope (AFM) further indicated that the new proposed method is a non-destructive method that does not induce structural changes during sample preparation.

Molecular cloning, expression, and characterization of BmSOD3 in silkworm (Bombyx mori).

Superoxide dismutases (SODs) play an essential role in eliminating excess reactive oxygen species and maintaining the redox balance of the immune system. To study the function of BmSOD3 in silkworm, 543-bp full-length complementary DNA-encoding BmSOD3 was cloned from silkworm. The BmSOD3 amino acids were compared to their homologs, and several highly conserved regions were analyzed. We also carried out phylogenetic analyses of the SOD gene. Our results showed that the BmSOD3 gene belonged with the ecCu/Zn SOD gene. The BmSOD3 gene was transformed into the pET28a vector for functional expression in Escherichia coli. The sodium salt-polyacrylamide gel electrophoresis results showed that the molecular weight of recombinant BmSOD3 was about 22 kDa. The recombinant protein BmSOD3 was purified to detect its properties. After purification analyses, the enzyme activity showed Cu/Zn SOD activity, and the specific activity of the purified enzyme was 0.51 U/mg. The BmSOD3 transcripts showed tissue-specific expression in the midgut and malpighian tubule. The immune microarray data for BmSOD3 showed an expression signal that had a strong response to the induction of four pathogens (Bacillus bombyseptieus, Beauveria bassiana, E. coli, and nuclear polyhedrosis virus), particularly after infection for 24 h, which indicates that the BmSOD3 gene plays a key role in response to bacterial, fungal, and viral invasion. The fusion protein also showed antibacterial activity against E. coli in vitro. Thus, the fusion protein BmSOD3 exhibits antibacterial activity and may be used in production to combat diseases caused by bacteria in silkworm.

[Alterations of regional homogeneity in patients with psychogenic erectile dysfunction: A study by resting-state functional MRI].

OBJECTIVE: To study the correlation between the brain regional homogeneity (ReHo) features and the clinical characteristics of the patients with psychogenic erectile dysfunction (pED). METHODS: Using IIEF-5 and the Self-Esteem and Relationship (SEAR) questionnaire, we evaluated the erectile function and psychosocial status of 32 pED patients and 28 healthy male subjects. Then, we compared the regional brain activity between the patients and healthy controls by resting-state functional magnetic resonance imaging (RS-fMRI) and the ReHo method, analyzed the correlation of the ReHo value of the altered brain regions with the results of IIEF-5 and SEAR questionnaire investigation, and explored the relationship between the ReHo features and the symptoms of the pED patients. RESULTS: Compared with the healthy male subjects, the pED patients obtained significantly lower IIEF-5 scores (22.21 ± 0.98 vs 13.97 ± 3.60, P < 0.01) and SEAR scores (61.92 ± 3.73 vs 37.58 ± 7.96, P < 0.01), a higher ReHo value of the left lateral cerebellum, and a lower ReHo value of the right precentral gyrus. The ReHo value of the left lateral cerebellum was correlated negatively with the IIEF-5 scores (r= －0.51, P < 0.01) and SEAR scores (r = －0.54, P < 0.01), while that of the right precentral gyrus positively with the IIEF-5 scores (r = 0.57, P < 0.01) and SEAR scores (r = 0.66, P < 0.01). CONCLUSIONS: Patients with pED had lateral cerebellum-mediated abnormal sensory integration and precentral gyrus-related dysfunction of motor imagery and motor execution.

Effect of Shading and Forest Type on Morphological Characteristics and Bioactive Compounds of Fruiting Bodies of Songshan Lingzhi Ganoderma tsugae (Agaricomycetes).

Ganoderma tsugae strain MCCCMAS0053 cultivation on short logs results in varied yield and quality under different growth conditions. Thus, growth conditions need optimization to increase yield and quality. An indoor experiment with three shade treatments (A1, two layers of black sun-shade net; A2, one layer of black sun-shade net plus plastic mulch; A3, one layer of black sun-shade net) and a field experiment in two forest types (pine or mixed pine-oak) were conducted. The results showed that shading and forest type significantly affected light intensities and the growth, size, biomass, and bioactive components of G. tsugae fruiting bodies. In the indoor experiment, the mean dry weight of the fruiting body and the diameter of the pileus in A2 increased by 21.51-44.98% and 13.42-22.26%, respectively, compared with those of A1 and A3. Similarly, the accumulation of the bioactive compounds (polysaccharides, total amino acids, and total essential amino acids) in the pileus and stipe were greater in A2 than in A1 and A3. Furthermore, compared with pure pine forest cultivation, fruiting bodies cultivated in mixed pine-oak forest had greater dry weight, pileus diameter, and more bioactive compounds. In addition, no significant difference was found between the A2 and mixed pine-oak treatments, which had a similar light intensity (from 1116 to 2367 lx). Hence, this suggests that the A2 shade treatment or cultivation in mixed pine-oak forest is beneficial for production of G. tsugae fruiting bodies, and light intensity may play a critical role in this process.

Intracellular symbionts drive sex ratio in the whitefly by facilitating fertilization and provisioning of B vitamins.

Symbionts can regulate animal reproduction in multiple ways, but the underlying physiological and biochemical mechanisms remain largely unknown. The presence of multiple lineages of maternally inherited, intracellular symbionts (the primary and secondary symbionts) in terrestrial arthropods is widespread in nature. However, the biological, metabolic, and evolutionary role of co-resident secondary symbionts for hosts is poorly understood. The bacterial symbionts Hamiltonella and Arsenophonus have very high prevalence in two globally important pests, the whiteflies Bemisia tabaci and Trialeurodes vaporariorum, respectively. Both symbionts coexist with the primary symbiont Portiera in the same host cell (bacteriocyte) and are maternally transmitted. We found that elimination of both Hamiltonella and Arsenophonous by antibiotic treatment reduced the percentage of female offspring in whiteflies. Microsatellite genotyping and cytogenetic analysis revealed that symbiont deficiency inhibited fertilization in whiteflies, leading to more haploid males with one maternal allele, which is consistent with distorted sex ratio in whiteflies. Quantification of essential amino acids and B vitamins in whiteflies indicated that symbiont deficiency reduced B vitamin levels, and dietary B vitamin supplementation rescued fitness of whiteflies. This study, for the first time, conclusively demonstrates that these two intracellular symbionts affect sex ratios in their whitefly hosts by regulating fertilization and supplying B vitamins. Our results reveal that both symbionts have the convergent function of regulating reproduction in phylogenetically-distant whitefly species. The 100% frequency, the inability of whiteflies to develop normally without their symbiont, and rescue with B vitamins suggests that both symbionts may be better considered co-primary symbionts.

Biotin provisioning by horizontally transferred genes from bacteria confers animal fitness benefits.

Insect symbionts are widespread in nature and lateral gene transfer is prevalent in insect symbiosis. However, the function of horizontally transferred genes (HTGs) in insect symbiosis remains speculative, including the mechanism that enables insects to feed on plant phloem deficient in B vitamins. Previously, we found there is redundancy in biotin synthesis pathways from both whitefly Bemisia tabaci and symbiotic Hamiltonella due to the presence of whitefly HTGs. Here, we demonstrate that elimination of Hamiltonella decreased biotin levels but elevated the expression of horizontally transferred biotin genes in whiteflies. HTGs proteins exhibit specific expression patterns in specialized insect cells called bacteriocytes housing symbionts. Complementation with whitefly HTGs rescued E. coli biotin gene knockout mutants. Furthermore, silencing whitefly HTGs in Hamiltonella-infected whiteflies reduced biotin levels and hindered adult survival and fecundity, which was partially rescued by biotin supplementation. Each of horizontally transferred biotin genes are conserved in various laboratory cultures and species of whiteflies with geographically diverse distributions, which shares an evolutionary origin. We provide the first experimental evidence that biotin synthesized through acquired HTGs is important in whiteflies and may be as well in other animals. Our findings suggest that B vitamin provisioning in animal-microbe symbiosis frequently evolved from bacterial symbionts to animal hosts through horizontal gene transfer events. This study will also shed light on how the animal genomes evolve through functional transfer of genes with bacterial origin in the wider contexts of microbial ecology.