In Vitro Bioactivities and Characterization of Mycelial Extracts from Different Strains of Phellinus igniarius (Agaricomycetes).

To fully utilize Phellinus igniarius fermentation mycelia, the present study investigated the in vitro antioxidant and α-amylase inhibitory properties of four Ph. igniarius strains. Organic solvents were used to extract fatty acids, phenolics, and flavonoids from the selected mushrooms. The composition and bioactivity of the extracts were evaluated. The lipid yield obtained using petroleum ether (7.1%) was higher than that obtained using 1:1 n-hex-ane+methanol (5.5%) or 2:1 dichloromethane+methanol (3.3%). The composition and relative content of saturated and unsaturated fatty acids in the petroleum ether extract were higher than those in other solvent extracts. Furthermore, ethyl acetate extracts had higher flavonoid and phenolic content and better antioxidant activity than other extracts; however, the 70% ethanol extracts had the best α-amylase inhibitory activity. The supernatant from the ethanol precipitation of aqueous and 1% (NH4)2C2O4 extracts could also be biocompound sources. This comparative study is the first highlighting the in vitro antioxidant and α-amylase inhibitory properties of the four strains of Ph. igniarius extracts prepared using different organic solvents, which makes the investigated species and extracts promising for biological application.

First Report of Sphingobacterium multivorum Causing Bacterial Rot of Cauliflower mushroom (Sparassis latifolia) in China.

Cauliflower mushroom (Sparassis latifolia), is widely distributed in Australia, North America, Europe, and East Asia (Bashir et al., 2020). It is known for its medicinal significance due to the availability of various pharmacological substances and their use in health supplements (Bashir et al., 2017). In recent years, with the development of artificial cultivation technology, S. latifolia has been industrialized in China, with an annual output value 50 million dollars. In March 2023, approximately 15% of S. latifolia showed obvious bacterial rot in mushroom hothouse (about 0.05 ha), located in Shuangliu county, Sichuan province, China (104°7'51"N, 30°25'2"E). The affected parts appear water-soaked, and become sunken and softened as the disease progresses. In the finally, all the fruiting body tissues turn into paste, with colors pale yellow, and have a foul smell. The pathogen was isolated from the margin of the lesions by dilution and streaking techniques onto Nutrient Agar, and incubated at 28℃ in the dark for 2-3 days. A single colony was re-streak for purification. Eight isolates were obtained from five samples collected randomly. The representative three isolates were selected for further characterization. For pathogenicity testing, ten health fruit bodies of S. latifolia were selected (for per isolate). Bacterial suspensions (1 × 107 CFU/ml) of the three isolates were applied to the fruiting body until wet, sterile water was used as controls. All the S. latifolia were maintained at 19±1℃, 85-100% relative humidity, and 18 h of light in the mushroom hothouse. Three days later, the inoculated fruiting bodies developed yellow color, and appear water-soaked, five days later, fruiting body gradually turn to soft and part turn to rot, seven days later, the fruiting body tissues completely turn into paste with a foul smell. The symptoms exhibited were similar to those of the original diseased fruiting bodies, while the control group remained healthy. The same bacterial were re-isolated from the infected fruiting bodies and subsequently identified by morphological characteristics and DNA sequenced. The pathogenicity test was conducted three times, each yielding similar results. The colonies of the pathogen are gram-negative rods, medium sized, convex, smooth, opaque, turning yellow after several days at a temperature 28℃. For molecular identification, the DNA of the representative three isolates was extracted using a Bacterial Genomic DNA Extraction Kit (Solarbio, Beijing). The 16S rRNA genes were amplified and sequenced with the primer 27F/1492R (Lane et al., 1985). Finally, the sequences were identical. The generated representative sequence was deposited in GenBank with accession number OR399122. BLASTn analysis showed 100% identity (1404/1404 bp) with previously deposited sequence (accession number CP068224) of S. multivorum FDAARGOS in GenBank. Based on the maximum likelihood method, phylogenetic analysis revealed 100% bootstrap support values with S. multivorum. Finally, the bacterium was identified as S. multivorum. This is the first report of S. multivorum causing bacterial rot of mushroom. The fruiting body of S. multivorum consists of multiple folded flat lobes, which are thin and have large surface area, may facilitate the infection of S. multivorum. Sphingobacterium sp. are named for their synthesize sphingolipids, which play an important role in bacterial infection (Kunz et al., 2019). These results will contribute to developing control strategies for this disease.

Enriched Environment Inhibits Neurotoxic Reactive Astrocytes via JAK2-STAT3 to Promote Glutamatergic Synaptogenesis and Cognitive Improvement in Chronic Cerebral Hypoperfusion Rats.

Chronic cerebral hypoperfusion (CCH) is a primary contributor to cognitive decline in the elderly. Enriched environment (EE) is proved to improve cognitive function. However, mechanisms involved remain unclear. The purpose of the study was exploring the mechanisms of EE in alleviating cognitive deficit in rats with CCH. To create a rat model of CCH, 2-vessel occlusion (2-VO) surgery was performed. All rats lived in standard or enriched environments for 4 weeks. Cognitive function was assessed using the novel object recognition test and Morris water maze test. The protein levels of glutamatergic synapses, neurotoxic reactive astrocytes, reactive microglia, and JAK2-STAT3 signaling pathway were measured using Western blot. The mRNA levels of synaptic regulatory factors, C1q, TNF-α, and IL-1α were identified using quantitative PCR. Immunofluorescence was used to detect glutamatergic synapses, neurotoxic reactive astrocytes, and reactive microglia, as well as the expression of p-STAT3 in astrocytes in the hippocampus. The results demonstrated that the EE mitigated cognitive impairment in rats with CCH and enhanced glutamatergic synaptogenesis. EE also inhibited the activation of neurotoxic reactive astrocytes. Moreover, EE downregulated microglial activation, levels of C1q, TNF-α and IL-1α and phosphorylation of JAK2 and STAT3. Our results suggest that inhibition of neurotoxic reactive astrocytes may be one of the mechanisms by which EE promotes glutamatergic synaptogenesis and improves cognitive function in rats with CCH. The downregulation of reactive microglia and JAK2-STAT3 signaling pathway may be involved in this process.

Enhanced Heat Resistance in Morchella eximia by Atmospheric and Room Temperature Plasma.

This study focuses on optimizing the mutagenesis process for Morchella eximia (Mel-7) mycelia through atmospheric and room temperature plasma (ARTP) mutation and explores the resultant thermal adaptability and physiological responses of mutant strains. This research demonstrated a clear relationship between ARTP mutagenesis exposure duration and lethality rate, indicating that an exposure time of 40 s resulted in the optimal balance of inducing mutations without causing excessive mortality. Additionally, this study established 43 °C as the ideal screening temperature for identifying mutant strains with enhanced heat resistance, as this temperature significantly challenges the mycelia while allowing thermotolerant strains to be distinguishable. Among the screened mutants, strains L21, L23, L44, and L47 exhibited superior growth and high-temperature tolerance, with notable resilience at 30 °C, highlighting their enhanced adaptability to above-optimal temperatures. Furthermore, this research delved into biochemical responses, including lipid peroxidation and non-enzymatic antioxidant content, highlighting the diverse mechanisms, such as enhanced lipid peroxidation resistance and increased antioxidant content, employed by mutant strains to adapt to temperature fluctuations. The activities of antioxidant enzymes, including peroxidase (POD) and superoxide dismutase (SOD), were shown to be significantly influenced by temperature elevations, illustrating their critical roles in the thermal adaptation of mutant strains. These findings shed light on the importance of considering mutation duration and temperature screening in the development of thermotolerant fungal strains with potential applications in various industries. This study's breakthrough lies in its comprehensive understanding of the thermal adaptability of Mel-7 mycelia and the identification of promising mutant strains, offering valuable insights for both academic and industrial purposes.

First Report of Lecanicillium aphanocladii Causing Rot of Sparassis crispa in China.

Sparassis crispa, also known as cauliflower mushroom, is a new popularly edible mushroom in China, also a medicinal mushroom, which possesses various biological activities, such as immunopotentiation, anti-diabetes, anti-cancer, and anti-inflammatory effects. (Han et al., 2018). In recent years, the artificial cultivation of S. crispa has gained considerable public attention in China. In 2023, approximately 20% of S. crispa (about 0.05 ha of the planting area) showed obvious rot with white molds symptoms in mushroom hothouse, located in Shuangliu county, Sichuan province, China (GPS, 104°7'51"N, 30°25'2"E). Infected fruiting bodies were covered by white mycelia that later turned red or fuchsia. In the final stages of infection, the S. crispa fruiting bodies turned dark red or brown before rotting. The pathogen was isolated from the margin of the lesions by plating onto potato dextrose agar (PDA), and incubated at 25℃ in the dark for a week. Five pure culture fungal isolates were obtained. Collected isolates with similar morphology were described as Lecanicillium spp. (Zare et al., 2001). The colonies were raised, covered with white, the reverse side were violet brown, produced diffusing reddish-purple pigment. Conidiogenous cells produced singly, in pairs, verticillate or in dense irregular clusters on prostrate hyphae, at first flask-shaped, tapering into threadlike neck, with a size of 3.0-6.2×0.8-2.2 µm. Conidia were solitary, oval to subglobose, and 2.3-4.0×1.1-2.1 µm in size, similar to L. aphanocladii (Higo et al., 2021). For pathogenicity testing, ten fruiting bodies of S. crispa (planted in the bottles) were selected. Fungal cake of the isolate Bx-Ljb of L. aphanocladii were applied to the fruiting body of S. crispa, whereas pieces of sterile PDA medium were used as controls. All the bottles were incubated at 19±1℃, 85-100% relative humidity, and 18 h of light in the mushroom hothouse. A week later, the inoculated fruiting bodies developed brown spots and gradually expanding, with symptoms similar to the original diseased fruiting bodies. The controls remained healthy. The same fungus was reisolated from the infected fruiting bodies and subsequently identified by morphological characteristics and DNA sequence analysis. The pathogenicity test was repeated three times with similar results. For molecular identification, the DNA of the isolates was extracted using a Fungi Genomic DNA Extraction kit (Solarbio, Beijing). The SSU, LSU, and TEF1-α genes were amplified with the primer as previously described (Zhou et al., 2018). The generated sequences were deposited in GenBank with accession numbers OR206377, OR206378, and OR204702, respectively. BLASTn analyses showed >99.2% identity with previously deposited sequences of L. aphanocladii. Based on the maximum likelihood method, phylogenetic analysis revealed 99% bootstrap support values with L. aphanocladii. The fungus was identified as L. aphanocladii based on morphological and multilocus phylogenetic analyses. To our knowledge, there are two reports of L. aphanocladii on fruiting bodies of Tremella fuciformis and Morchella sextelata in China, and this is the first report of this fungus causing rot of S. crispa in China. It may be a reminder that the risk of L. aphanocladii in mushroom production in China is gradually increasing. These results will contribute to developing managemental strategies for this disease in S. crispa.

Evaluation of the nutritional value, umami taste, and volatile organic compounds of Hypsizygus marmoreus by simulated salivary digestion in vitro.

Hypsizygus marmoreus is an edible medicinal mushroom species with a high dietary value. The main purpose of this study was to evaluate the nutritional value, umami taste, and volatile organic compounds (VOCs) of H. marmoreus treated with hot water combined with simulated salivary digestion in vitro. Seafood mushroom (Hm3) had the highest content of moisture, soluble polysaccharides, soluble proteins, and total flavonoids while white Hypsizygus marmoreus (Hm1) had the highest total phenolic content. Moreover, Hm1 had a more noticeable equivalent umami concentration (EUC) value, indicating the umami properties of Hm1 as a food or processing ingredient. Results from E-nose and HS-SPME-GC-MS revealed that the VOCs of Hm1 and brown Hypsizygus marmoreus (Hm2) were relatively similar, which differed substantially from Hm3. Among the 134 VOCs, 24 differential metabolites were identified by OPLS-DA analysis, characterized by VIP > 1, p-value < 0.05, and FC > 2 (pairwise comparisons). Furthermore, 10 biomarkers with VIP > 1 and p-value < 0.05 were identified by PLS-DA analysis based on the total differential metabolites to distinguish different strains of H. marmoreus. These results will benefit future research on the chemistry of H. marmoreus and serve as a guide for breeding, introducing, and using the species more effectively.

Application of Atmospheric and Room-Temperature Plasma (ARTP) to Microbial Breeding.

Atmospheric and room-temperature plasma (ARTP) is an efficient microbial mutagenesis method with broad application prospects. Compared to traditional methods, ARTP technology can more effectively induce DNA damage and generate stable mutant strains. It is characterized by its simplicity, cost-effectiveness, and avoidance of hazardous chemicals, presenting a vast potential for application. The ARTP technology is widely used in bacterial, fungal, and microalgal mutagenesis for increasing productivity and improving characteristics. In conclusion, ARTP technology holds significant promise in the field of microbial breeding. Through ARTP technology, we can create mutant strains with specific genetic traits and improved performance, thereby increasing yield, improving quality, and meeting market demands. The field of microbial breeding will witness further innovation and progress with continuous refinement and optimization of ARTP technology.

Combined transcriptomics and metabolomics analysis reveals the molecular mechanism of heat tolerance of Le023M, a mutant in Lentinulaedodes.

Lentinula edodes, one of the most highly regarded edible mushrooms in China, is susceptible to damage from high temperatures. However, a mutant strain derived from L. edodes, known as Le023M, has shown exceptional thermotolerance. Compared to the original strain Le023, Le023M exhibited accelerated mycelial recovery following heat stress. Through RNA-seq analysis, the majority of differentially expressed genes (DEGs) were found to be associated with functions such as "protein refolding", "protein unfolding", "protein folding", and "response to heat", all of which are closely linked to heat shock proteins. Furthermore, qRT-PCR results revealed significant accumulation of heat shock-related genes in Le023M under heat stress. GC-MS analysis indicated elevated levels of trehalose, aspartate, and glutamate in Le023M when subjected to heat stress. The highly expressed genes involved in these metabolic pathways were predominantly found in Le023M. Collectively, these findings highlight the following: (i) the crucial role of heat shock proteins (HSPs) in the thermo-resistant mechanisms of Le023M; (ii) the potential of trehalose accumulation in Le023M to enhance mycelium resistance to heat stress; and (iii) the induction of aspartate and glutamate accumulation in response to heat stress. These results shed light on the molecular mechanisms underlying the thermotolerance of Le023M, providing valuable insights for further understanding and improving heat stress response in L. edodes. The findings also highlight the potential applications of Le023M in the cultivation and production of L. edodes under high-temperature conditions.

First Report of Leaf blight Caused by Colletotrichum karstii on Banana Shrub (Michelia figo) in Sichuan, China.

Banana Shrub (Michelia figo (Lour.) Spreng.) is widely cultivated in most of southern China (Wu et al, 2008). It can be used to make essential oil and flower tea（Ma et al, 2012; Li et al, 2010）.The first symptoms were observed in Sept. 2020 at a grower's field in Banana shrub seedlings (0.6 ha), Ya'an city (29°30'N, 102°38'E), Hanyuan county. The symptoms re-occurred in May-June of 2021 and became prevalent from August to September. the incidence rate and the disease index were 40% and 22%, respectively. Initially, purplish-brown necrotic lesions appeared at the leaf tip with dark-brown edges. Progressively, necrosis spread, to the middle of the leaves, and the older area turned gray-white. Dark sunken lesions appeared in the necrotic areas and orange conidial masses were visible under humid conditions. Ten isolates were obtained on potato dextrose agar (PDA) from 10 leaf samples using previously described tissue isolation method (Fang et al. 1998). All the 10 isolates exhibited similar morphological characteristics. Grey to white aerial mycelium at the center and in dispersed tufts, with numerous dark conidiomata scattered over the surface, reverse was pale orange with numerous dark flecks corresponding to the ascomata, orange conidial masses were formed from mature conidiomata. Conidia were hyaline, smooth-walled, aseptate, straight, cylindrical, apex round, the contents appearing granular 14.8 to 17.2 × 4.2 to 6.4 µm (average: 16.26 × 4.84 µm, n=30) as Colletotrichum spp. (Damm et al. 2012). For molecular identification, DNA was extracted from a representative isolate HXcjA using a plant genomic DNA extraction kit (Solarbio, Beijing). and the partial sequences of internal transcribed spacer region (ITS, OQ641677), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, OL614009), actin (ACT, OL614007), beta-tubulin (TUB2, OL614011), histone3 (HIS3, OL614010), and calmodulin (CAL, OL614008) were amplified and sequenced using the primer pairs ITS1/ITS4 (White et al. 1990), GDF/GDR (Templeton et al. 1992), ACT-512F/ACT-783R, CAL 228F/CAL 737R (Carbone et al. 1999), TUB1F/Bt2bR, CYLH3F/CYLH3R (Crous et al. 2004), respectively. BLASTn analysis for ITS, GAPDH, CAL, ACT, TUB2 and HIS3 sequences showed ≥99.7% identity to C. Karstii, namely, NR_144790 (532/532 bp), MK963048 (252/252 bp), MK390726 (431/431 bp), MG602039 (761/763 bp), (KJ954424, 294/294 bp), (KJ813519, 389/389 bp), respectively. The fungus was identified as C. karstii based on morphology and a multigene phylogeny. The conidial suspension (1 × 107 conidia/mL) with 0.05% Tween 80 buffer was used for pathogenicity test, by spraying 2-year-old Banana Shrub plants. Ten plants were inoculated with spore suspensions (approximately 2ml per plant). An equal number of plants were sprayed with 0.05% Tween 80 buffer to serve as a control. Fifteen days later, the inoculated plants showed similar symptoms as the original diseased plants but the controls remained asymptomatic. C. karstii was re-isolated from the infected leaves and identified by morphology and a multigene phylogeny. The pathogenicity test was repeated three times with similar results, confirming Koch's postulates. To our knowledge, this is the first report of Banana Shrub leaf blight caused by C. karstii in China. This disease reduces the ornamental and economic value of Banana Shrub, and this work will provide a basis for the prevention and treatment of the disease in the future.

Disturbed functional connectivity and topological properties of the frontal lobe in minimally conscious state based on resting-state fNIRS.

Background: Patients in minimally conscious state (MCS) exist measurable evidence of consciousness. The frontal lobe is a crucial part of the brain that encodes abstract information and is closely related to the conscious state. We hypothesized that the disturbance of the frontal functional network exists in MCS patients. Methods: We collected the resting-state functional near-infrared spectroscopy (fNIRS) data of fifteen MCS patients and sixteen age- and gender-matched healthy controls (HC). The Coma Recovery Scale-Revised (CRS-R) scale of MCS patients was also composed. The topology of the frontal functional network was analyzed in two groups. Results: Compared with HC, the MCS patients showed widely disrupted functional connectivity in the frontal lobe, especially in the frontopolar area and right dorsolateral prefrontal cortex. Moreover, the MCS patients displayed lower clustering coefficient, global efficiency, local efficiency, and higher characteristic path length. In addition, the nodal clustering coefficient and nodal local efficiency in the left frontopolar area and right dorsolateral prefrontal cortex were significantly reduced in MCS patients. Furthermore, the nodal clustering coefficient and nodal local efficiency in the right dorsolateral prefrontal cortex were positively correlated to auditory subscale scores. Conclusion: This study reveals that MCS patients' frontal functional network is synergistically dysfunctional. And the balance between information separation and integration in the frontal lobe is broken, especially the local information transmission in the prefrontal cortex. These findings help us to understand the pathological mechanism of MCS patients better.

Application of omics technology in the research on edible fungi.

Edible fungus is a large fungus distributed all over the world and used as food and medicine. But people's understanding of edible fungi is not as much as that of ordinary crops, so people have started a number of research on edible fungi in recent years. With the development of science and technology, omics technology has gradually walked into people's vision. Omics technology has high sensitivity and wide application range, which is favored by researchers. The application of omics technology to edible fungus research is a major breakthrough, which has transferred edible fungus research from artificial cultivation to basic research. Now omics technology in edible fungi has been flexibly combined with other research methods, involving multiple studies of edible fungus, such as genetic breeding, growth and development, stress resistance, and the use of special components in edible fungus as pharmaceutical additives. It is believed that in the future, the research of edible fungi will also be brought to a deeper level with the help of omics technology. This paper introduces the application progress of modern omics technology to the study on edible fungi and mentions the application prospect of edible fungi research with the constant development of omics technology, thereby providing ideas for the follow-up in-depth research on edible fungi.

Genome-Wide Identification and Analysis of the Heat-Shock Protein Gene in L. edodes and Expression Pattern Analysis under Heat Shock.

Lentinula edodes (L. edodes), one of the most popular edible mushrooms in China, is adversely affected by high temperature. Heat shock proteins (HSPs) play a crucial role in regulating the defense responses against the abiotic stresses in L. edodes. Some HSPs in L. edodes have been described previously, but a genome-wide analysis of these proteins is still lacking. Here, the HSP genes across the entire genome of the L. edodes mushroom were identified. The 34 LeHSP genes were subsequently classified into six subfamilies according to their molecular weights and the phylogenetic analysis. Sequence analysis showed that LeHSP proteins from the same subfamily have conserved domains and one to five similar motifs. Except for Chr 5 and 9, 34 LeHSPs genes were distributed on the other eight chromosomes. Three pairs of paralogs were identified because of sequence alignment and were confirmed as arising from segmental duplication. In LeHSPs' promoters, different numbers of heat shock elements (HSEs) were predicted. The expression profiles of LeHSPs in 18N44 and 18 suggested that the thermo-tolerance of strain 18N44 might be related to high levels of LeHSPs transcript in response to heat stress. The quantitative real-time PCR (qRT-PCR) analysis of the 16 LeHSP genes in strains Le015 and Le027 verified their stress-inducible expression patterns under heat stress. Therefore, these comprehensive findings provide useful in-depth information on the evolution and function of LeHSPs and lay a theoretical foundation in breeding thermotolerant L. edodes varieties.

Edible and medicinal fungi breeding techniques, a review: Current status and future prospects.

Mushrooms of the edible and medicinal which are highly nutritious and environmentally friendly crops carry numerous medicinal benefits. For the abundant and high diversity of bioactive metabolites they possess, which are considered to be an important pool of bioresources. The efficient breeding technique is always a challenging task in mushrooms for obtaining better character strains, which are essential for developing healthy products and even consumption. This review comprehensively summarizes the breeding techniques applied to the edible and medicinal mushrooms. Including the traditional mutagenesis method, and even modern gene-editing breeding techniques, the effects of each method, and the comparison of each breeding technique are systematic illustrations. Strategies for mushroom breeding techniques in the future are also discussed in this review paper. With the ongoing sequencing of the mushroom genome, knowledge of the gene background of the strains and functions can be available for developing better markers for gene-editing breeding as CRISPR/Cas9 systems. Combine the metabolism engineering and in-silico tools analysis was the rational design of the novel strains. Modern physical mutagenesis techniques such as the ARTP and the combination of the other physical, and chemical breeding mutagens with cross-breeding techniques or the protoplasts fusion will also lead to superior strains for cultivation and pave the way for higher quality and yield.

The resting-state topological organization damage of language-related brain regions in post-stroke cognitive impairment.

The topology of brain networks is the foundation of cognition. We hypothesized that stroke damaged topological organization resulting in cognitive impairment. The aim was to explore the damage pattern of the resting-state topology in post-stroke cognitive impairment (PSCI) patients. Thirty-seven patients with PSCI and thirty-seven gender- and age-matched healthy controls (HC) were recruited. The structural and functional data were collected from all subjects. The degree centrality (DC), betweenness centrality (BC), and global properties of brain networks were analyzed between groups. Spearman correlation analysis was performed between topological properties that changed significantly and clinical cognitive function scale scores. Compared with HC, the PSCI patients had significantly reduced DC in language-related brain regions and significantly higher DC in the right frontal lobe, hippocampus, and paracentral lobule. The decreased BC was located in the left caudate, thalamus, temporal, and frontal lobes. The increased BC was detected in the left cuneus and right precuneus. In addition, PSCI exhibited increased characteristic path length and decreased small-worldness. PSCI patients had impaired functional topology of the language-related brain regions, mainly in the left hemisphere. The enhanced processing and relaying information of some right high-order cognitive brain regions may be a compensatory mechanism. However, the whole brain's function integration was reduced, and there was an imbalance between efficiency and consumption.

Physical Exercise-Induced Astrocytic Neuroprotection and Cognitive Improvement Through Primary Cilia and Mitogen-Activated Protein Kinases Pathway in Rats With Chronic Cerebral Hypoperfusion.

Chronic cerebral hypoperfusion (CCH) is closely related to vascular cognitive impairment and dementia (VCID) and Alzheimer's disease (AD). The neuroinflammation involving astrocytes is an important pathogenic mechanism. Along with the advancement of the concept and technology of astrocytic biology, the astrocytes have been increasingly regarded as the key contributors to neurological diseases. It is well known that physical exercise can improve cognitive function. As a safe and effective non-drug treatment, physical exercise has attracted continuous interests in neurological research. In this study, we explored the effects of physical exercise on the response of reactive astrocytes, and its role and mechanism in CCH-induced cognitive impairment. A rat CCH model was established by 2 vessel occlusion (2VO) and the wheel running exercise was used as the intervention. The cognitive function of rats was evaluated by morris water maze and novel object recognition test. The phenotypic polarization and the primary cilia expression of astrocytes were detected by immunofluorescence staining. The activation of MAPKs cascades, including ERK, JNK, and P38 signaling pathways, were detected by western blot. The results showed that physical exercise improved cognitive function of rats 2 months after 2VO, reduced the number of C3/GFAP-positive neurotoxic astrocytes, promoted the expression of S100A10/GFAP-positive neuroprotective astrocytes, and enhanced primary ciliogenesis. Additionally, physical exercise also alleviated the phosphorylation of ERK and JNK proteins induced by CCH. These results indicate that physical exercise can improve the cognitive function of rats with CCH possible by promoting primary ciliogenesis and neuroprotective function of astrocytes. The MAPKs signaling cascade, especially ERK and JNK signaling pathways may be involved in this process.

Transcriptome Profiling Reveals Candidate Genes Related to Stipe Gradient Elongation of Flammulina filiformis.

Stipe gradient elongation is an important and remarkable feature in the development of most mushroom fruiting bodies. However, its molecular mechanism has rarely been described. Here, the decreasing trend of stipe elongation and increasing trend of cell length in a gradient from the top to the base of the stipe were determined in a model basidiomycete mushroom: Flammulina filiformis. According to RNA-seq results, 1409 differentially expressed genes (DEGs) were identified among elongation region (ER), transition region (TR), and stable region (SR) samples, including 26 transcription factors (TFs). Based on Short Time-series Expression Miner (STEM) clustering of DEGs, clusters 1 and 3, with obvious expression trends that were consistent with or in contrast to the elongation rate, were screened. The cluster 1 DEGs were mainly involved in the GO cellular component category and KEGG genetic information processing class; however, the cluster 3 DEGs were mainly involved in metabolic processes. Furthermore, qRT-PCR confirmed that key genes of the long-chain fatty acid synthesis pathway were involved in stipe gradient elongation and regulated by NADPH oxidase-derived ROS signaling molecules. These findings provide an essential basis for understanding the molecular mechanism of stipe gradient elongation.

Effect of Tetramethylpyrazine on Neuroplasticity after Transient Focal Cerebral Ischemia Reperfusion in Rats.

Tetramethylpyrazine (TMP) has been widely used in ischemic stroke in China. The regulation of neuroplasticity may underlie the recovery of some neurological functions in ischemic stroke. Middle cerebral artery occlusion (MCAO) model was established in this study. Rats were divided into three groups: sham group, model group, and TMP group. The neurological function was evaluated using modified neurological severity score (mNSS). Following the neurological function test, expression of synaptophysin (SYP) and growth-associated protein 43 (GAP-43) were analyzed through immunohistochemistry at 3 d, 7 d, 14 d, and 28 d after MCAO. Finally, the synaptic structural plasticity was investigated using transmission electron microscopy (TEM). The TMP group showed better neurological function comparing to the model group. SYP levels increased gradually in ischemic penumbra (IP) in the model group and could be enhanced by TMP treatment at 7 d, 14 d, and 28 d, whereas GAP-43 levels increased from 3 d to 7 d and thereafter decreased gradually from 14 d to 28 d in the model group, which showed no significant improvement in the TMP group. The results of TEM showed a flatter synaptic interface, a thinner postsynaptic density (PSD), and a wider synaptic cleft in the model group, and the first two alterations could be ameliorated by TMP. Then, a Pearson's correlation test revealed mNSS markedly correlated with SYP and synaptic ultrastructures. Taken together, TMP is capable of promoting functional outcome after ischemic stroke, and the mechanisms may be partially associated with regulation of neuroplasticity.

Effect of enriched environment on angiogenesis and neurological functions in rats with focal cerebral ischemia.

The purpose of this study was to investigate the effect of enriched environment (EE) on cerebral angiogenesis after ischemia-reperfusion injury. Middle cerebral artery occlusion (MCAO) followed by reperfusion was performed in rats to set up an animal model of ischemia-reperfusion injury. In a set of behavioral tests, we demonstrated that the animals in the IEE (ischemia + enriched environment) group exhibited significantly improved neurological functions compared to those in the standard housing condition group. In consistent with the functional tests, smaller infarction volumes were observed in the animals of IEE group. Laser scanning confocal microscopy and 3D quantitative analysis of cerebral microvessels revealed that EE treatment increased the total vessel surface area and number of branch point in the ischemic boundary zone. IgG extraction assay showed that the blood brain barrier (BBB) leakage in the ischemic brain was attenuated after EE treatment. EE treatment also enhanced endothelial cells (ECs) proliferation and increased the expression levels of VEGF and its receptor Flk-1 after ischemia-reperfusion injury. Analyses of Spearman's correlation coefficients indicated a correlation of mNSS scores with enhanced cerebral angiogenesis. Together, the results suggest that EE treatment-induced cerebral angiogenesis may contribute to the improved neurological outcome of stroke animals after ischemia-reperfusion injury.

Effects of Tetramethylpyrazine on Functional Recovery and Neuronal Dendritic Plasticity after Experimental Stroke.

The 2,3,5,6-tetramethylpyrazine (TMP) has been widely used in the treatment of ischemic stroke by Chinese doctors. Here, we report the effects of TMP on functional recovery and dendritic plasticity after ischemic stroke. A classical model of middle cerebral artery occlusion (MCAO) was established in this study. The rats were assigned into 3 groups: sham group (sham operated rats treated with saline), model group (MCAO rats treated with saline) and TMP group (MCAO rats treated with 20 mg/kg/d TMP). The neurological function test of animals was evaluated using the modified neurological severity score (mNSS) at 3 d, 7 d, and 14 d after MCAO. Animals were euthanized for immunohistochemical labeling to measure MAP-2 levels in the peri-infarct area. Golgi-Cox staining was performed to test effect of TMP on dendritic plasticity at 14 d after MCAO. TMP significantly improved neurological function at 7 d and 14 d after ischemia, increased MAP-2 level at 14 d after ischemia, and enhanced spine density of basilar dendrites. TMP failed to affect the spine density of apical dendrites and the total dendritic length. Data analyses indicate that there was significant negative correlation between mNSS and plasticity measured at 14 d after MCAO. Thus, enhanced dendritic plasticity contributes to TMP-elicited functional recovery after ischemic stroke.