In order to detect how miRNA-mRNA interaction participates in drought response and drought memory ofA. squarrosum, the drought-sensitive ecotype AEX ofAgriophyllum squarrosumwas selected as the plant material, and transcriptome and sRNA sequencing were performed after three rounds of drought-rehydration treatments. A total of 341 and 7858A. squarrosummiRNAs (DEMIs) and mRNAs (DEMs) were identified in response to drought stress, respectively. The DEMI-DEM regulatory network showed that 24A. squarrosumDEMIs were involved in the regulation of 1423 DEMs, and these target genes were mainly involved in cell differentiation function. Two drought memory miRNAs (DMMIs), miR156 and miR6173, were identified, which regulated 11 and 6 drought memory genes (DMGs), respectively. Target genes for miR156 included three transcription factors (TFs):AsRWP-RK,AsC3H11, andAsWRKY33; the target genes for miR6173 included twoGSTs (glutathione S-transferase). In previous study,AsC3H11andAsWRKY33participated in theA. squarrosumdrought memory hierarchical regulatory network as TF molecular switches, soA. squarrosummiR156 could serve as a candidate hub for the “miRNA-transcription factor-functional gene” network. This study partially elucidated the molecular regulatory basis of drought response and drought memory inA. squarrosum, which will provide a theoretical basis for the development of stress-tolerant genetic resources in desert plants.

In order to investigate whether Gly33 residue in the conserved sequence of the active site of human thioredoxin hTrx-1 is essential in the process of reducing oxidative protein, the Gly33 residue of hTrx-1 was deleted or replaced (G→A) by two synthetic mutant genes. The synthetic mutant genes were then expressed in prokaryotic cells, further purified and tested for antioxidant activity. The results showed that both the deletion of Gly33 residue and the mutation to Ala, the mutated hTrx-1 could be efficiently expressed in prokaryotic cells, indicating that both mutant hTrx-1 could form a “Trx fold” structure. Antioxidant activity testing showed that in the absence of Gly33 residues, its antioxidant capacity decreased, when Gly33 mutated to Ala, its antioxidant capacity significantly decreased to near loss. Therefore, this article speculated that the change of conserved amino acid residues in the Trx-1 active center affects the geometric configuration of the active center region, resulting in difficulty in approaching the substrate protein in space. The detection results of the content of disulfide bond sulfhydryl groups involved in the formation of disulfide bonds showed that the content of disulfide bond sulfhydryl groups formed in mutant hTrx-1 was significantly lower than that in wild-type hTrx-1. It was speculated that the ability of Cys32 and Cys35 to form disulfide bonds in mutant hTrx-1 was affected.

To evaluate the copy number ofCtDXRgene in transgenicNicotiana benthamianaand further analyze its tolerance to salt and heat stress, Southern Blot was applied to determine the copy number of theActingene in wild-typeN. benthamianaand the copy number of theCtDXRgene inCtDXRtransgenicN. benthamianawas detected by real-time PCR withActingene as reference gene. Then, the salt and heat tolerance ofCtDXRtransgenicN. benthamianawas analyzed. According to the PCR-dependent method, 10 of T1 transgenicN. benthamianawere randomly detected and all of which obtained positive results, indicating thatCtDXRhas been successfully transformed. TheActingene is a single-copy gene in the genome of wild-typeN. benthamiana. 80% of transgenicN.benthamianawere determined as single-copy or low-copy lines.CtDXRtransgenicN. benthamianashowed better performance, such as plant height (P<0.01), number of lateral roots (P<0.01), and fresh weight (P<0.001), than wild-typeN. benthamianaunder salt stress, whereasCtDXRtransgenicN. benthamianaexhibited better performance, such as plant height (P<0.01), number of leaves (P<0.01), and fresh weight, than wild-typeN. benthamianaunder heat stress. Taken together,CtDXRconferred salt and heat tolerance toN.benthamiana. In this study, a method to detect copy number of foreign gene in transgenicN. benthamianawas established, by which the copy number ofCtDXRgene in transgenicN. benthamianawas accomplished. Moreover,CtDXRgene was preliminarily identified to have salt and heat tolerance.

To construct a B lymphocyte model stably expressing human TLR2, the coding sequences of humanTLR1,TLR2andTLR6were cloned and inserted into the pCDH-CMV-MCS-EF1-GFP-puro lentiviral vector. After verification of correct insertion by sequencing, these plasmids were packaged into complete infectious virus particles by transfection into 293T cells and transduced into Nalm-6 cells (TLR2－), and puromycin was used to select Nalm-6 cells that stably expressed TLR2 (TLR2+). An inverted fluorescence microscope was used to observe the cell status and green fluorescent protein expression. Western blott was used to measure the expression levels of related proteins. CCK-8 and flow cytometry assays were used to determine the cell viability and evaluate cell proliferation. The results of inverted fluorescence microscopy showed that Nalm-6 cells were successfully infected with lentiviruses with green fluorescent protein expression and puromycin resistance. Western Blot analysis showed that TLR2 was successfully expressed in Nalm-6 cells and that activation of TLR2 signaling significantly increased the levels of phosphorylated PI3K-AKT signaling axis proteins. The CCK-8 and flow cytometry assays showed that TLR2 activation could increase the cell viability and promote cell proliferation. In summary, in this study, a B lymphocyte line model overexpressing TLR2 was successfully constructed and was used to verify that TLR2 activation can upregulate the PI3K-AKT signaling pathway in B cells and promote their proliferation. This model lays the foundation for further exploring the impact of the TLR2 pathway on the anti-infective immune function of B cells.

The aim of this study was to investigate the influence of auraptene (AUR) on food intake in mice and its mechanism. Mice were randomly divided to three groups including control group, 2 μmol/kg AUR group and 1 μmol/kg AUR group. AUR was continuously given by oral gavage for 2 weeks to measure the cumulative food intake of the mice, the expression of insulin-like growth factors (IGFs) in serum, liver and hypothalamus, insulin-like growth factor-binding proteins (IGFBPs) in liver, proopiomelanocortin (POMC) in hypothalamus. The levels of mRNA and protein of carnitine palmitoyltransferase 1A (CPT1A) as a peroxisome proliferator-activated receptor α (PPARα) target gene were also tested. The results showed that AUR promoted food intake in mice in a concentration-dependent manner, inhibited the expression of POMC (P<0.01), significantly promoted the expression of IGFBP-1 in liver (P<0.01), and significantly reduced the content of IGF-1 in serum and hypothalamus (P<0.01), but did not effect on IGF-1 in liver (P>0.05). Meanwhile, AUR significantly increased the expression of CPT1A in liver (P<0.01). In summary, these findings suggested that AUR could significantly promote food intake in mice. The main mechanism was that AUR promoted food intake in mice because of increasing the expression of IGFBP-1 in liver by PPARα activation, which bound the free IGF-1 in serum to decrease IGF-1 in hypothalamus and inhibit the expression of POMC.

NewFlammulina velutipes-based composite material was prepared by freeze-drying method withFlammulina velutipesrich in chitin and dextran as raw material after extraction of polysaccharides. TheFlammulina velutipes-based composite was analyzed by infrared spectrum and scanning electron microscope. The samples characteristics such as anti-microbial properties, mechanical properties, swelling property, water vapor permeability, coagulation performancein vitro, degradability and cytotoxicity were assayed. Results showed thatFlammulina velutipessamples had excellent antibacterial properties, and the bacteriostatic rates ofEscherichia coliandStaphylococcus aureuswere 98.51% and 88.18% respectively.Flammulina velutipes-based composite dressing had excellent physical properties and high softness, and had excellent water absorption and water retention. Its breaking strength was 15.20 MPa and elongation was 84.11%. Its water vapor permeability number reached 2500 g/(m2·d), the coagulation indexin vitrowas 0.3. Moreover, the dressing was non-toxic and harmless, and the soil degradation rate could reach 79.79% after 35 days of discarding. This study demonstrated thatFlammulina velutipes-based composite dressing had excellent medical potential and could be used in wound dressing and tissue engineering. This study was of great significance to promote the utilization of by-products of fungal polysaccharide extraction and enhance the added value ofFlammulina velutipes.

To obtain halophilic cellulase-producing fungi from the Chaka Salt Lake, Qinghai Province, PDA medium containing 5% NaCl was used as an isolation medium to obtain halophilic fungi strains. After initial screening with sodium carboxymethyl cellulose (CMC) as the only carbon source, the enzyme properties of cellulase produced by high-activity strains were studied by Congo red plate. The results showed that 24 strains of cellulose-degrading bacteria were obtained from the samples of Chaka Salt Lake, which belonged to 10 different genera. A highly active strain was obtained by primary screening and rescreening, named DaliF28, and identified asCladosporiumsp. by ITS gene. After 32 hours of liquid fermentation and culture ofCladosporiumsp. DaliF28, the cellulase activity in the supernatant reached a maximum of 32.87 U/mL. The optimum temperature, pH, and salt concentration of cellulases produced byCladosporiumsp. DaliF28 were 50 ℃, 6.6, and 2.5 mol/L NaCl, respectively. It remained above 60% relative activity after incubated at the optimum pH and salt concentration at 60 ℃ for 1 h, and still exhibited more than 60% relative activity at the concentration of 0－4 mol/L NaCl. The isolated strains indicated that there are abundant halophilic cellulase-producing fungi in Chaka Salt Lake.

45 single colonies with certain morphological characteristics were isolated from saline-alkali soil in northern Ningxia, and the 16S rRNA sequence of the strains were sequenced and identified by genome extraction technology. Results showed that there were 16 strains of actinomycetes, including 13 strains of Streptomyces, 3 strains of Acidophilum. The experimental results of antibacterial activity exhibited that all 16 strains of actinomycetes had antibacterial activity, among which 12 strains of secondary metabolites had significant antibacterial activity againstProteus vulgaris, 12 strains of secondary metabolites had obvious inhibitory activity againstStaphylococcus aureus, and 9 strains of secondary metabolites had certain growth inhibitory activity againstCandida albicans. The researches demonstrated that the microbial strains isolated from saline-alkali soil in northern Ningxia had strong antibacterial or bacteriostatic activity. This study not only enriched the microbial resources in special habitats, but also laid a foundation for further isolation of antibacterial lead compounds.

UsingHordeum bogdaniiseeds with endophytic fungi (E+) and without endophytic fungi (E－) as materials, seedlings were planted in greenhouses at first and then transplanted in fields with different salinity and alkalinity. The surface soils sampled in spring, summer and autumn, and spring samples as control groups, were used to determine the activities of catalase, urease, sucrase and alkaline phosphatase, as well as the contents of organic carbon, total nitrogen, total phosphorus, total potassium, available phosphorus and available potassium. The results showed that the contents of urease, sucrase, alkaline phosphatase, organic carbon, nitrogen, and potassium were the highest under moderate salt stress. With the increase of planting time ofH. bogdanii, the contents of catalase, carbon, nitrogen, phosphorus, potassium, and available potassium had no significant change. The contents of urease, sucrase, and available phosphorus increased, and the activity of alkaline phosphatase increased under medium salt. Moreover, the soil enzyme activity and the contents of carbon, nitrogen, phosphorus, potassium, available phosphorus, and available potassium in E+H. bogdaniiwere mostly higher than those in E－ under different salt levels. This indicated that endophytic fungi infection could improve the enzyme activity and soil nutrients of near plants by regulating them, laying a certain foundation for further research on the impact of endophytic fungi expansion on soil microbial diversity inH. bogdanii.

The bio-reactor is a key process and important limiting factor during the treatment of livestock and poultry wastewater through microalgal bacterial biofilm. In this paper, a biofilm reactor was designed for microalgal bacterial consortia to treat piggery wastewater, and the influent ammonia nitrogen mass concentration was adjusted to explore its impact on nutrients removal and microalgae biomass production. Moreover, scanning electron microscopy was used to observe the formation of microalgal bacterial biofilm on the carrier. High-throughput sequencing was also used for microbial community analysis. The aim of this study was to reveal the change regularities of nutrients removal and the response mechanism of microbial community of microalgal bacterial biofilm wastewater treatment with the fluctuation of influent ammonia. The results showed that the maximum daily removal rate of ammonia nitrogen could reach 35.60 mg/（L·d） when the influent ammonia nitrogen mass concentration was 200 mg/L, and the dominant bacterial genus wasPrevotella_ 9,Clostridium_ Sensu_ Stricto_ 1,AcinetobacterandBifidobacterium. Moreover, a large number of microalgal bacterial clusters were observed on carrier illustrating the formation of microalgal bacterial film. When the influent ammonia nitrogen mass concentration increased to 300 mg/L, the daily removal rate of ammonia nitrogen significantly increased reaching a maximum of 45.70 mg/（L·d）. However, the sustained high ammonia nitrogen load made a certain impact on the microalgal bacterial biofilm, while the dominant bacterial genera turned toComamonasandChristensenellaceae_ R-7_ Group.

In order to study the effect of salt stress on the growth and development ofPlatycodon grandiflorus, the experiment was conducted with two-year-oldPlatycodon grandiflorusas test material, four treatment groups including control (CK, normal watering), low concentration salt stress (S1, 200 mmol/L NaCl), medium concentration salt stress (S2, 400 mmol/L NaCl) and high concentration salt stress (S3, 600 mmol/L NaCl) were set up, the biomass, physiological and biochemical indexes and total saponins content ofPlatycodon grandiflorusseedlings were determined. The results showed that salt stress inhibited the growth ofPlatycodon grandiflorusand reduced its biomass with the increase of salt stress concentration, and S1 could promote the increase of chlorophyll content ofPlatycodon grandiflorus, S2 and S3 decreased the content of chlorophyll, the relative water content of leaves decreased with the increase of salt stress concentration, the activities of antioxidant enzymes in leaves and roots ofPlatycodon grandiflorusdecreased significantly at S3 and the 30th day after treatment. The contents of proline, MDA and total saponins in leaves and roots ofPlatycodon grandiflorusincreased significantly with the increase of salt stress concentration and treatment time. This study can provide a theoretical basis for improving the growth, yield and quality ofPlatycodon grandiflorusin salt-alkali soil.

In order to investigate the viability of kiwifruit pollen, and to study the morphological changes before and after pollen germination, 15 samples ofActinidia deliciosapollens were used in this experiment. Anin vitroculture method was used to determine pollen germination viability, and scanning electron microscopy was used to observe the morphology of five female pollens and ten male pollens before and after germination. The results showed that the germination rate of pollens from male plants varied considerably. None of the five female pollens germinated. Among the 10 male pollen samples No. 8 had the highest germination rate of 79.27% and No. 9 had the lowest germination rate of 22.85%. The polar facial view of the 15 samples did not change before and afterex vivoincubation. These samples were all trilobate circles. However, there was a significant difference in their equatorial view. The pollen morphology did not change before and after the female plants were cultured in isolation, and they were all spheroidal. The pollens of the male plant were perprolate or prolate before germination, and after germination in culturein vitro, all 10 samples were sub-spherical, with the polar axis length becoming almost smaller, the equatorial axis length becoming larger, the P/E ratio becoming smaller, and the width of the germination groove becoming larger. The differences in the values of the morphological indexes between the samples were small.

It was found that there were calcium phosphate crystals inMammillaria schumannii. The tubercles ofM. schumanniiwere observed using a scanning electron microscope and an X-ray energy spectrum. The results showed that the mineral crystals, contained the O, Ca, and P elements, which were calcium phosphate and mainly embedded in the cortical cell wall with sporadic distribution in the cells. The majority of their morphological features resemble bouquets dispersed in all directions or just one, and rarely, olive balls coated with membranes could be seen. After weak-light treatment inM. schumannii, it was found that the crystal density in tubercles decreased obviously, a little amount of calcium phosphate crystals gathered together, and the overall size also decreased significantly (P<0.001). The results here provided a better understanding of the morphological characteristics of plant calcium phosphate as well as their roles in plants.

In order to investigate the mechanism of NF-κB in enhancing CyHV-2 replication, transcriptional and translational levels of CyHV-2 genes at 24, 72 and 96 h post CyHV-2 infection in the presence of NF-κB agonist were monitored by RT- qPCR and Western Blot assays.The results showed that the transcriptional levels and protein levels of CyHV-2 genes were increased at 72 and 96 h in the presence of NF-κB agonist.To further elucidate how NF-κB affects the transcription and translation of CyHV-2, the binding sites of the promoter region for NF-κB were predicted and dual luciferase reporter gene experiments were performed to study the regulatory role of NF-κB on viral gene expression.The experimental results showed that NF-κB overexpression increased CyHV-2 promoter activity, further suggesting that NF-κB promotes viral replication by activating the promoter of CyHV-2 genes.This study should provide novel strategies for anti-CyHV-2 drugs and enhance the understanding of CyHV-2 replication.

In this work, the research progress of brain-derived neurotrophic factor (BDNF) in exercise to improve cognitive dysfunction in autism spectrum disorder (ASD) was reviewed. It focused on the biological functions of BDNF and its involvement in the upstream SIRT1-PGC1a/FNDC5 pathway and downstream Src/PLC-γ1, Ras/MAPK, and PI3K/Akt signaling pathways, alterations in brain regions associated with cognitive function in ASD patients, as well as the mechanisms of BDNF expression and regulation in different pathological biological models of ASD. The paper elaborated on the potential role of BDNF in improving synaptic plasticity, learning and memory abilities, and ultimately cognitive functions of the hippocampus in the brain. In this way, the role of BDNF in promoting neuronal synaptic plasticity, improving learning and memory abilities, and ultimately improving cognitive functions in the brain could be explored, and the regulation of BDNF expression might play an important role in improving hippocampal cognitive functions in patients with ASD, which might provide a new perspective for understanding the complex causal relationship between down-regulation of BDNF expression and the improvement of hippocampal cognitive functions of patients with autism through exercise. It would provide a new perspective for understanding the complex causal relationship between BDNF expression and the improvement of hippocampal cognitive function in autistic patients through exercise. Finally, it was expected that the targeting of BDNF will provide a broad prospect for the development of biomarkers and drugs for the clinical diagnosis of autism, a strong theoretical basis for the clinical treatment of ASD, and new ideas and theoretical foundations for exercise rehabilitation.

Cyanobacteria are mostly distributed in oceans, salt lakes and hypersaline environments. Due to natural selection and genetic differentiation in salt environments, Cyanobacteria have formed special survival mechanisms including in unique community structure composition, diversity distribution and intracellular secondary metabolites. Currently, the diversity, the dominant genus, the salt adaptation mechanism, the species distribution and application of Cyanobacteria need to be further explored from different salt environments. This article systematically reviewed the recent advances in the population diversity, the community structure compositions and the adaptive survival mechanisms(e.g. accumulation of compatible solutes or antioxidants, balance of intracellular ions, and/or increase of antioxidant enzyme activity) of Cyanobacteria among various salt environments. Further, the representative species of intracellular secondary metabolites (e.g. polyketones, oligopeptides/polypeptides, alkaloids, lipids, terpenes and polysaccharides), chemical structure types and main bioactive functions (e.g. antibacterial, anti-inflammatory, antitumor, antimalarial and protease inhibition) were also systematically summarized. It probably provided a reference for subsequent salina algae mining and the utilization of natural metabolites.

This study is to establish a real-time fluorescence detection method for rapidly detecting the hepatitis B virus using enzyme recombinase amplification (ERA) technology. The primers and probes were designed according to the conserved sequence of the polymerase coding region (P region) of the hepatitis B virus, and the reaction conditions, such as primers, temperature, and fluorescent probe concentration, were optimized to establish the optimal reaction system of the ERA real-time fluorescence assay, which was further validated in terms of sensitivity, specificity, anti-jamming capacity, and clinical samples. The results showed that the ERA real-time fluorescence method could detect the presence of the hepatitis B virus within 20 minutes at 42 ℃, with a limit of detection of 102copies/μL. The other four viruses (hepatitis A virus, hepatitis C virus, Epstein-Barr virus, and Cytomegalovirus) did not show fluorescence amplification curves, indicating reasonable specificity and anti-interference ability. When compared to the real-time fluorescence PCR method, the ERA real-time fluorescence method for 58 hepatitis B virus clinical samples exhibited a sensitivity of 97.37%, a specificity of 100%, a negative predictive value of 95.24%, a positive predictive value of 100%, and Kappa value of 0.96. This study successfully established a simple, rapid, sensitive, specific, and low-cost method for early detection of hepatitis B virus to meet the need for rapid detection.

Based on the concept of “integration of science and education” and “outcome based education (OBE)”, the frontier scientific and technological achievements in the field of biochemistry were introduced into teaching experiments to cultivate students’ scientific research and innovation quality. The scientific research project “the effects of preparation methods on the properties of microbial polysaccharides” was introduced through the whole teaching process. Firstly, the students were guided to retrieve the literature and discuss the hot issues of “the development of a new type of environmentally friendly oil-displacing agent that was resistant to temperature and salt” to broaden their professional vision and cultivate their literature retrieving ability. Secondly, students were encouraged to design individualized experimental schemes according to their own knowledge system to cultivate their ability to analyze and solve problems. Thirdly, they prepared polysaccharide samples by salt-alcohol-precipitation method and Sevage method respectively, calculated the yield, characterized the samples by infrared spectra and ultraviolet spectra, and investigated the influence of different preparation methods on visco-elasticity and displacement effect of polysaccharides. This process helped students to establish their multidisciplinary knowledge system of “biology-chemical engineering-energy” and improved their experimental practice ability. Finally, the students carried out data analysis, program evaluation and achievement roadshow to train their scientific research ability.

The course of “fundamentals of plant cloning technology and applications” is a general course offered by college of life science of Zhejiang University for all undergraduates. The course covers culture-medium preparation, aseptic manipulation, induction of callus and embryoid, acclimation of tissue culture seedlings, which embodies the cross-integration of biology, forestry and horticulture. This paper explores the cross-fusion knowledge points between the curriculum and other professional fields from the aspects of curriculum teaching objectives, curriculum system construction and curriculum assessment methods. Based on the problem-oriented teaching methods and the combination of online and offline teaching methods, the role of curriculum teaching content and teaching methods in cultivating interdisciplinary students is explored. The results of three years’ teaching practice show that the students’ academic performance, interest in scientific research and experiment, and curriculum construction level have all been improved. The interdisciplinary teaching design has enhanced students’ serial cognition of interdisciplinary knowledge, expanded students’ horizons, and laid a good foundation for the cultivation of innovative talents with multi-disciplinary background.

Traditional medical practice courses have disadvantages, including a lack of initiative in learning or cross-disciplinary teaching cooperation. Therefore, it was necessary to adopt new medical remote-interactive teaching models to meet the needs of Internet times. Anesthesiology, as an essential part of medicine, is based on the theories and techniques of biomedicine, neurobiology, and clinical medicine. Taking the practical anesthesiology course as the cornerstone to build a “campus-hospital” learning community, various teaching methods, such as problem-based learning (PBL), case-based learning (CBL), and scenario simulation, were integrated during the teaching practice. In addition, the way of assessment was also changed. The new remote interactive teaching mode cultivated students’ ability to independent learning and innovative spirit and improved the students’ overall quality.