Experimental Verification of Erchen Decoction Plus Huiyanzhuyu Decoction in the Treatment of Laryngeal Squamous Cell Carcinoma Based on Network Pharmacology.

BACKGROUND: The prescription of Chinese herbal medicine (CHM) consists of multiple herbs that exhibit synergistic effects due to the presence of multiple components targeting various pathways. In clinical practice, the combination of Erchen decoction and Huiyanzhuyu decoction (EHD) has shown promising outcomes in treating patients with laryngeal squamous cell carcinoma (LSCC). However, the underlying mechanism by which EHD exerts its therapeutic effects in LSCC remains unknown. METHODS: Online databases were utilized for the analysis and prediction of the active constituents, targets, and key pathways associated with EHD in the treatment of LSCC. The protein-protein interaction (PPI) network of common targets was constructed and visualized using Cytoscape 3.8.1 software. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to investigate the functional roles of core targets within the PPI network. Protein clustering was conducted utilizing the MCODE plug-in. The obtained results highlight the principal targets and pathways involved. Subsequently, clinical samples were collected to validate alterations in the levels of these main targets through Western blotting (WB) and immunohistochemistry (IHC). Furthermore, both in vivo and in vitro experiments were conducted to investigate the therapeutic effects of EHD on healing LSCC and elucidate its underlying mechanism. Additionally, to ensure experimental reliability and reproducibility, quality control measures utilizing HPLC were implemented for EHD herbal medicine. RESULTS: The retrieval and analysis of databases in EHD medicine and LSCC disease yielded a total of 116 overlapping targets. The MCODE plug-in methods were utilized to acquire 8 distinct protein clusters through protein clustering. The findings indicated that both the first and second clusters exhibited a size greater than 6 scores, with key genes PI3K and ErbB occupying central positions, while the third and fourth clusters were associated with proteins in the PI3K, STAT3, and Foxo pathways. GO functional analysis reported that these targets had associations mainly with the pathway of p53 mediated DNA damage and negative regulation of cell cycle in terms of biological function; the death-induced signaling complex in terms of cell function; transcription factor binding and protein kinase activity in terms of molecular function. The KEGG enrichment analysis demonstrated that these targets were correlated with several signaling pathways, including PI3K-Akt, FoxO, and ErbB2 signaling pathway. On one hand, we observed higher levels of key genes such as P-STAT3, P-PDK1, P-Akt, PI3K, and ErbB2 in LSCC tumor tissues compared to adjacent tissues. Conversely, FOXO3a expression was lower in LSCC tumor tissues. On the other hand, the key genes mentioned above were also highly expressed in both LSCC xenograft nude mice tumors and LSCC cell lines, while FOXO3a was underexpressed. In LSCC xenograft nude mice models, EHD treatment resulted in downregulation of P-STAT3, P-PDK1, PI3K, P-AKT, and ErbB2 protein levels but upregulated FOXO3a protein level. EHD also affected the levels of P-STAT3, P-PDK1, PI3K, P-AKT, FOXO3a, and ErbB2 proteins in vitro: it inhibited P-STAT3, P-AKT, and ErbB2, while promoting FOXO3a; however, it had no effect on PDK1 protein. In addition, HPLC identified twelve compounds accounting for more than 30% within EHD. The findings from this study can serve as valuable guidance for future experimental investigations. CONCLUSION: The possible mechanism of EHD medicine action on LSCC disease is speculated to be closely associated with the ErbB2/PI3K/AKT/FOXO3a signaling pathway.

Recombinase polymerase amplification combined with Pyrococcus furiosus Argonaute for fast Salmonella spp. testing in food safety.

Foodborne illness caused by Salmonella spp. is one of the most prevalent public health problems globally, which have brought immeasurable economic burden and social impact to countries around the world. Neither current nucleic acid amplification detection method nor standard culture method (2-3 days) are suitable for field detection in areas with a heavy burden of Salmonella spp. Here, we developed a highly sensitive and accurate assay for Salmonella spp. detection in less than 40 min. Specifically, the invA gene of Salmonella spp. was amplified by recombinase polymerase amplification (RPA), followed by Pyrococcus furiosus Argonaute (PfAgo)-based target sequence cleavage, which could be observed by a fluorescence reader or the naked eye. The assay offered the lowest detectable concentration of 1.05 × 101 colony forming units/mL (CFU/mL). This assay had strong specificity and high sensitivity for the detection of Salmonella spp. in field samples, which indicated the feasibility of this assay.

Effects of collagen-based coating with chitosan and ε-polylysine on sensory, texture, and biochemical changes of refrigerated Nemipterus virgatus fillets.

In order to evaluate the effects of chitosan, ε-polylysine, and collagen on the preservation properties of refrigerated Nemipterus virgatus, samples were tested with different treatments for 10 days, namely chitosan, ε-polylysine and collagen (CH + ε-PL + CA), chitosan and ε-polylysine (CH + ε-PL), chitosan and collagen (CH + CA), ε-polylysine and collagen (ε-PL + CA), and the uncoated sample (CK). The results demonstrated that the bio-coating exhibited better preservation effects. The CH + ε-PL + CA, CH + ε-PL, CH + CA, ε-PL + CA treatments could significantly inhibit bacterial growth and retard the increase of total volatile base nitrogen (TVB-N), 2-thiobarbituric acid (TBA), K-value, and total viable counts (TVC) in N. virgatus fillets. The pH of all samples decreased and reached its lowest value on day 6, then increased significantly at the end of the experiment (p < .05). Water-holding capacity (WHC) of all the groups decreased continuously throughout storage, and CK reached 66.03% on day 6, which is significantly lower than CH + ε-PL + CA, CH + ε-PL, CH + CA, and ε-PL + CA (p < .05). On the contrary, the sensory scores of CH + ε-PL + CA, CH + ε-PL, CH + CA, and ε-PL + CA were significantly higher than the control, and the score of CH + ε-PL + CA (p < .05) was the best among all the groups. In terms of texture, CH + PL + CA also showed less cell shrinkage and tighter muscle fiber arrangement compared to other treatments. To sum up, the CH + PL + CA bio-coating proved to be a promising method for maintaining the storage quality of N. virgatus under refrigerated storage conditions.

Design and development of a rapid meat detection system based on RPA-CRISPR/Cas12a-LFD.

In recent years, meat adulteration safety incidents have occurred frequently, triggering widespread attention and discussion. Although there are a variety of meat quality identification methods, conventional assays require high standards for personnel and experimental conditions and are not suitable for on-site testing. Therefore, there is an urgent need for a rapid, sensitive, high specificity and high sensitivity on-site meat detection method. This study is the first to apply RPA combined with CRISPR/Cas12a technology to the field of multiple meat identification. The system developed by parameter optimization can achieve specific detection of chicken, duck, beef, pork and lamb with a minimum target sequence copy number as low as 1 × 100 copies/µL for 60 min at a constant temperature. LFD test results can be directly observed with the naked eye, with the characteristics of fast, portable and simple operation, which is extremely in line with current needs. In conclusion, the meat identification RPA-CRISPR/Cas12a-LFD system established in this study has shown promising applications in the field of meat detection, with a profound impact on meat quality, and provides a model for other food safety control programs.

Orally administered Chinese herbal therapy to assist post-surgical recovery for chronic rhinosinusitis-A systematic review and meta-analysis.

This systematic review and meta-analysis aims to: assess the effectiveness and safety of orally administered Chinese herbal medicines (CHMs) as adjuncts to the post-surgical management of chronic rhinosinusitis (CRS); inform clinicians of the current evidence; identify the best available evidence; and suggest directions for further research. Randomised controlled trials (RCTs) were identified from searches of nine databases plus clinical trial registries. Participants were adults and/or children diagnosed with sinusitis or rhinosinusitis, with or without nasal polyps, who had received surgery. Interventions were CHMs used orally following surgery for CRS as additions to conventional post-surgical management. Controls received conventional post-surgical management without CHMs. Studies reported results for Sino-Nasal Outcome Test (SNOT), visual analogue scales (VAS), Lund-Mackay computed tomography score (LM), Lund-Kennedy endoscopic score (LK), mucociliary transport time (MTT), mucociliary transport rate (MTR), mucociliary clearance (MC) or quality of life (QoL). Twenty-one RCTs were included. All used oral CHMs following functional endoscopic sinus surgery (FESS). The pooled results showed no significant difference between groups for SNOT-20 at the end of treatment (EoT) but there was a significant difference at follow up (FU) in favour of additional CHMs. The VAS for total nasal symptoms (VAS-TNS) showed greater improvements in the CHM groups at EoT and FU. Only FU data were reported for LM which showed greater improvement in the CHM groups. LK showed greater improvements at EoT and FU. The measures of mucociliary transport (MTT, MTR, and MC) each showed significantly greater improvement at EoT in the group that received additional CHMs. No study reported QoL. Adverse events were not serious, but reporting was incomplete. The meta-analyses suggested the addition of oral CHMs to conventional management following FESS may improve recovery. However, most studies were not blinded, and substantial heterogeneity was evident in some meta-analyses. Blinded studies are required to further investigate the roles of oral CHMs in post-surgical recovery. Systematic review registration number: The protocol was registered in PROSPERO (CRD42019119586).

Multidisciplinary strategies to enhance therapeutic effects of flavonoids from Epimedii Folium: Integration of herbal medicine, enzyme engineering, and nanotechnology.

Flavonoids such as baohuoside I and icaritin are the major active compounds in Epimedii Folium (EF) and possess excellent therapeutic effects on various diseases. Encouragingly, in 2022, icaritin soft capsules were approved to reach the market for the treatment of hepatocellular carcinoma (HCC) by National Medical Products Administration (NMPA) of China. Moreover, recent studies demonstrate that icaritin can serve as immune-modulating agent to exert anti-tumor effects. Nonetheless, both production efficiency and clinical applications of epimedium flavonoids have been restrained because of their low content, poor bioavailability, and unfavorable in vivo delivery efficiency. Recently, various strategies, including enzyme engineering and nanotechnology, have been developed to increase productivity and activity, improve delivery efficiency, and enhance therapeutic effects of epimedium flavonoids. In this review, the structure-activity relationship of epimedium flavonoids is described. Then, enzymatic engineering strategies for increasing the productivity of highly active baohuoside I and icaritin are discussed. The nanomedicines for overcoming in vivo delivery barriers and improving therapeutic effects of various diseases are summarized. Finally, the challenges and an outlook on clinical translation of epimedium flavonoids are proposed.

Light-driven textile sensors with potential application of UV detection.

Smart textiles based on monitoring systems of health conditions, structural behaviour, and external environmental conditions have been presented as elegant solutions for the increasing demands of health care. In this study, cotton fabrics (CFs) were modified by a common strategy with a dipping-padding procedure using reduced graphene oxide (RGO) and a photosensitive dye, spiropyran (SP), which can detect environmental UV light. The morphology of the CF is observed by scanning electron microscopy (SEM) measurements showing that the topography structure of coatings is related to the SP content. The resistance of the textile sensors decreases after UV radiation, which may be attributed to the easier electron transmission on the coatings of the CF. With the increase of SP content, the introduction of a large amount of SP within the composites could cause discontinuous distributions of RGO in the fiber surfaces, preventing electron transmission within the coatings of the RGO. The surface wettability of the coatings and the sweat sensitivity are also studied before and after UV radiation.

Chinese herbal therapy in the management of rhinosinusitis-A systematic review and meta-analysis.

This systematic review aims to assess the effects and safety of Chinese herbal medicines (CHMs) in the management of rhinosinusitis (RS); inform clinicians of the current state of the evidence; identify the best available evidence; and suggest further directions for research. Five English and four Chinese language databases, and four clinical trial registries were searched. Eligible studies were randomised controlled trials (RCTs). Participants were diagnosed with RS based on established criteria. Test interventions were CHMs administered orally and/or nasally, excluding injections and displacement techniques. Control interventions included placebos, no additional treatment, and conventional non-invasive treatments including pharmacotherapies and/or nasal irrigation, and/or inhalations. Polyposis and post-surgical recovery were excluded. Outcomes were Sino-Nasal Outcome Test (SNOT), visual analogue scales (VAS), Lund-Mackay computed tomography score (LM), Lund-Kennedy Endoscopic score (LK), Mucociliary transport time (MTT), Mucociliary transport rate (MTR), quality of life and adverse events (AEs). Risk of bias used the Cochrane tool. Meta-analysis in Review Manager 5.4.1 used random effects for mean difference (MD) or risk ratio (RR) with 95% confidence intervals. Heterogeneity was assessed as I2. Thirty-four RCTs were included, 30 of chronic RS (CRS) and four of acute RS (ARS). These enrolled 3,752 participants. Five RCTs blinded participants. For CRS, comparisons with placebo showed greater improvements in the CHM groups for SNOT-20 and VAS-TNS (total nasal symptoms). Blinded comparisons with pharmacotherapies showed no differences between groups in the degree of improvement for SNOT-20, VAS-TNS, and LM, suggesting these CHMs had similar effects, at least in the short term. In ARS, pooled results found improved scores on VAS-TNS and LK suggesting a benefit for combining these CHMs with pharmacotherapies. Limitations included inadequacies in study design and methodological reporting, and insufficient reporting of AEs. Heterogeneity in some pooled results precluded strong conclusions. Further well-designed studies are needed to test whether the results are replicable. Systematic review registration number: PROSPERO (CRD42019119586).

Construction of carbon-based flame retardant composite with reinforced and toughened property and its application in polylactic acid.

To simultaneously improve the flame retardancy, strength and toughness of polylactic acid (PLA) fibers, a composite flame retardant CNTs-H-C was prepared with carbon nanotubes (CNTs) as the core, hexachlorocyclotriphosphazene as linker, and chitosan grafted on the surface. The prepared CNTs-H-C was introduced into a PLA matrix to obtain CNTs-H-C/PLA composites and fibers via a melt-blending method. The morphology, structure, flame retardant properties and mechanical properties were thoroughly characterized, and the flame retardant mechanism was studied. Results showed that the prepared CNTs-H-C displayed a nanotube-like morphology with good compatibility and dispersion in the PLA matrix. After blending with PLA, CNTs-H-C/PLA composites exhibited outstanding flame retardancy with limiting oxygen index (LOI) increasing from 20.0% to 27.3%, UL94 rating reaching V-0. More importantly, the introduction of CNTs-H-C did not affect the spinnability of PLA. Compared with pure PLA fibers, the LOI of CNTs-H-C/PLA fibers with a CNTs-H-C content of 1.0 wt% increased by 32.5%, and meanwhile the breaking strength and elongation increased by 28.2% and 30.4%, respectively. Mechanism study revealed that CNTs-H-C/PLA possessed a typical condensed phase flame retardancy mechanism. In short, we have developed a CNT-based composite flame retardant with reinforced and toughened properties for the PLA matrix. The prepared CNTs-H-C showed great potential in polymer flame retardancy and mechanical enhancement.

Analysis of the Effect of Branched Chain Amino Acids on Muscle Health Information of Swimmers Based on Multisensor Fusion and Deep Learning.

Swimmers must fully mobilize the muscles of the whole body during exercise, and it is necessary to study the protection of swimmers from muscle damage. Now, muscle damage is increasing year by year, and more athletes are affected. Therefore, studying the causes of muscle injuries and exploring more effective treatments have become important research topics in the field of sports medicine. This study is mainly based on deep learning to analyze the protective effect of branched-chain amino acids on swimming athletes' muscle injury. Due to the complex and changeable environment and the interference of unknown factors, a single sensor cannot meet the needs of obtaining information. Therefore, people have developed the technology of multisensor information fusion to obtain enough information. Multisensor data fusion technology can synthesize the information of each sensor and then obtain more comprehensive and accurate decision-making information. This study is mainly based on multisensor fusion and deep learning to analyze the impact of branched chain amino acids on Swimmers' muscle health information. Finally, two experiments were designed in this article. The first experimental result showed that the pain level of the experimental group who took BCAA supplements was 19% lower than that of the control group that did not take the BCAA supplement within three days after exercise. The results of the second experiment show the following: after exercise, the creatine kinase activity value of the experimental group taking BCAA supplement was 4.38 ± 1.45, and the creatine kinase activity value of the control group taking placebo was 5.42 ± 2.12. It proves that BBCA can protect muscle damage by reducing the activity of creatine kinase.

A U-Box Type E3 Ubiquitin Ligase Prp19-Like Protein Negatively Regulates Lipid Accumulation and Cell Size in Chlamydomonas reinhardtii.

Microalgae lipid triacylglycerol is considered as a promising feedstock for national production of biofuels. A hotspot issue in the biodiesel study is to increase TAG content and productivity of microalgae. Precursor RNA processing protein (Prp19), which is the core component of eukaryotic RNA splice NTC (nineteen associated complex), plays important roles in the mRNA maturation process in eukaryotic cells, has a variety of functions in cell development, and is even directly involved in the biosynthesis of oil bodies in mouse. Nevertheless, its function in Chlamydomonas reinhardtii remains unknown. Here, transcriptional level of CrPrp19 under nutrition deprivation was analyzed, and both its RNA interference and overexpressed transformants were constructed. The expression level of CrPrp19 was suppressed by nitrogen or sulfur deficiency. Cell densities of CrPrp19 RNAi lines decreased, and their neutral lipid contents increased 1.33 and 1.34 times over those of controls. The cells of CrPrp19 RNAi lines were larger and more resistant to sodium acetate than control. Considerably none of the alterations in growth or neutral lipid contents was found in the CrPrp19 overexpression transformants than wild type. Fatty acids were also significantly increased in CrPrp19 RNAi transformants. Subcellular localization and yeast two-hybrid analysis showed that CrPrp19 was a nuclear protein, which might be involved in cell cycle regulation. In conclusion, CrPrp19 protein was necessary for negatively regulating lipid enrichment and cell size, but not stimulatory for lipid storage.

Preparation and properties of a novel poly(lactic-acid)-based thermoplastic vulcanizate from both experiments and simulations.

A novel bio-based thermoplastic vulcanizate (TPV) material consisting of poly(lactic acid) (PLA) and a novel polymeric slide ring material (SeRM) was fabricated via isocyanate-induced dynamic vulcanization. The microscopic morphology, thermal properties, biocompatibility, and mechanical properties of the SeRM/PLA TPV material were comprehensively investigated, in turn by transmission electron microscopy, differential scanning calorimetry, in vitro cytotoxicity test, electron tension machine, and molecular dynamics simulations. Phase inversion in TPV was observed during the dynamic vulcanization, and TEM images showed that SeRM particles that were dispersed in PLA continuous phase had an average diameter of 1-4 µm. Results also indicated that an optimum phase inversion morphology was obtained at the SeRM/PLA blending ratio of 70/30 w/w. Glass transition temperature of PLA was found to be slightly decreased, owing to the improvement in interface compatibility by chemically bonding the PCL side chains (of SeRM molecules) and PLA chains. The tensile strength and elongation at break of TPVs were approximately 14.7 MPa and 164%, respectively, at SeRM/PLA blending ratio of 70/30, owing to the unique sliding effect of SeRM molecules when subjected to deformations. Cytotoxicity test results proved that the bio-based TPVs were fully non-toxic to L929 cells. In such aspects we believe that the bio-based TPV can be a promising material in the biomedical applications as an alternative of traditional commodity plastics.

Efficacy and safety of the Chinese herbal medicine Xiao-qing-long-tang for allergic rhinitis: A systematic review and meta-analysis of randomized controlled trials.

ETHNOPHARMACOLOGICAL RELEVANCE: The classic Chinese herbal medicine formula Xiao-qing-long-tang (XQLT) is commonly recommended to manage allergic rhinitis (AR), but the treatment efficacy and safety of XQLT are uncertain. AIM OF THE STUDY: This study aimed to evaluate the effectiveness and safety of XQLT in treating AR. MATERIALS AND METHODS: Nine databases were searched from their inception to April 2021. Randomized controlled trials (RCTs) evaluating XQLT for AR were included. The methodological quality of the studies was assessed using the Cochrane risk-of-bias tool. A meta-analysis and a subgroup meta-analysis were conducted to evaluate the effectiveness of XQLT. RESULTS: Twenty-four RCTs were included in this meta-analysis. XQLT was compared to both placebo and Western medicine (WM), and XQLT combined with WM was compared with WM alone. Meta-analyses were conducted for total nasal symptom scores (TNSS), four individual nasal symptom scores, quality of life (QoL), effective rate, and recurrence rate. The TNSS decreased after XQLT treatment and combination treatment (mean difference (MD): -0.79; 95% confidence interval (CI) [-1.20, -0.38], standardized mean difference (SMD): -1.42; 95% CI [-1.59, -1.24], and SMD: -1.84; 95% CI [-2.08, -1.60]). The two individual nasal symptom scores decreased after XQLT treatment and combination treatment; these nasal symptoms comprised rhinorrhea (SMD: -0.30; 95% CI [-0.58, -0.02] and SMD: -0.48; 95% CI [-0.70, -0.26]), and nasal obstruction (SMD: -0.54; 95% CI [-0.78, -0.30] and SMD: -0.54; 95% CI [-0.76, -0.32). XQLT and XQLT combined with WM achieved a better effective rate than WM (risk ratio (RR): 1.18; 95% CI [1.11, 1.25] and RR: 1.16; 95% CI [1.10, 1.23]) and a lower recurrence rate than WM (RR: 0.24; 95% CI [0.13, 0.43] and RR: 0.47; 95% CI [0.31, 0.72]). XQLT was well tolerated in patients being treated for AR. CONCLUSION: Our results indicated that oral XQLT may alleviate the TNSS, rhinorrhea scores, and nasal obstruction scores of AR and is safe to use in clinical practice. However, more RCTs that follow rigorous methodologies and evaluate well-accepted outcome measures are required to evaluate the effectiveness of XQLT.

Comparison of physicochemical properties of different tissues from China climbing perch Anabas testudineus and crucian carp Carassius auratus.

This study aimed to investigate nutrition in climbing perch Anabas testudineus which is an important nutritious economic freshwater fish in Asia and compare with Carassius auratus (crucian carp). Three kinds of tissues, including muscle, livers, and eggs, were isolated, respectively. Physicochemical properties including moisture, ash, protein, amino acids, fat, vitamins, and calcium contents in those tissues were determined. The results showed climbing perch muscle and liver contained less moisture, but more protein, amino acids, and vitamins than crucian carp muscle and liver. While moisture, ash, protein, and total amino acids contents of climbing perch egg were lower than those of crucian carp egg. Climbing perch egg had more fat, vitamins, and calcium than crucian carp egg. The amino acid profile was also performed, and 16 amino acids were identified and quantified in muscle, liver, and egg. Among tissues, the highest and lowest concentration of total amino acid content was found in crucian carp eggs and livers, respectively. Glutamic acid (Glu) and histidine (His) were the most and least amino acids in climbing perch and crucian carp tissues, respectively. Sixteen amino acids in climbing perch egg were less than those in crucian carp egg while it is an opposite case in muscle and liver, which amino acids of climbing perch tissues were more than those of crucian carp muscle and liver. Vitamin A of climbing perch was more than crucian carp in all three tissues, but vitamin E content in climbing perch liver was lower than that of crucian carp liver. Calcium content of muscle had no difference between two species. The abovementioned comparison of physicochemical properties of different tissues from China climbing perch and crucian carp will provide a necessary supplementary of freshwater muscle nutrition research, also was helpful for application of climbing perch.

Screen-Printed Carbon Black/Recycled Sericin@Fabrics for Wearable Sensors to Monitor Sweat Loss.

Wearable sensors to monitor human sweat loss are important for real-time health monitoring, requiring electrically conductive, mechanically flexible fabrics as working electrodes. Here, a textile-based sweat monitor was fabricated by screen printing of carbon black and recycled sericin on cotton fabrics. The obtained fabric with excellent flexibility, good hydrophilicity (86°), and proper resistivity (61.7 Ω/cm2) can be used as a working electrode for a wearable sweat monitor. A wearable sweat monitor is highly sensitive (42.7% in acidic sweat), flexible, and can be washed (99.1% retention after 30 washes). This work offers a promising approach for the fabrication of wearable sensors and promotes the widespread applications of personalized health-monitoring devices.

Transcription Factors From Haematococcus pluvialis Involved in the Regulation of Astaxanthin Biosynthesis Under High Light-Sodium Acetate Stress.

The microalgae Haematococcus pluvialis attracts attention for its ability to accumulate astaxanthin up to its 4% dry weight under stress conditions, such as high light, salt stress, and nitrogen starvation. Previous researches indicated that the regulation of astaxanthin synthesis might happen at the transcriptional level. However, the transcription regulatory mechanism of astaxanthin synthesis is still unknown in H. pluvialis. Lacking studies on transcription factors (TFs) further hindered from discovering this mechanism. Hence, the transcriptome analysis of H. pluvialis under the high light-sodium acetate stress for 1.5 h was performed in this study, aiming to discover TFs and the regulation on astaxanthin synthesis. In total, 83,869 unigenes were obtained and annotated based on seven databases, including NR, NT, Kyoto Encyclopedia of Genes and Genomes Orthology, SwissProt, Pfam, Eukaryotic Orthologous Groups, and Gene Ontology. Moreover, 476 TFs belonging to 52 families were annotated by blasting against the PlantTFDB database. By comparing with the control group, 4,367 differentially expressed genes composing of 2,050 upregulated unigenes and 2,317 downregulated unigenes were identified. Most of them were involved in metabolic process, catalytic activity, single-organism process, single-organism cellular process, and single-organism metabolic process. Among them, 28 upregulated TFs and 41 downregulated TFs belonging to 27 TF families were found. The transcription analysis showed that TFs had different transcription modules responding to the high light and sodium acetate stress. Interestingly, six TFs belonging to MYB, MYB_related, NF-YC, Nin-like, and C3H families were found to be involved in the transcription regulation of 27 astaxanthin synthesis-related genes according to the regulatory network. Moreover, these TFs might affect astaxanthin synthesis by directly regulating CrtO, showing that CrtO was the hub gene in astaxanthin synthesis. The present study provided new insight into a global view of TFs and their correlations to astaxanthin synthesis in H. pluvialis.

Effects of embryonic exposure to bixafen on zebrafish (Danio rerio) retinal development.

Bixafen, a pyrazole-carboxamide fungicide, is a potent toxicant that may elicit multiple adverse effects in non-target organisms. However, knowledge of the mechanisms involved in developmental defects caused by bixafen in aquatic organisms remains limited. In this study, the effects of bixafen on retinal development were evaluated in embryo-larval zebrafish. We exposed zebrafish embryos to 0, 0.1, and 0.3 µM bixafen. Exposure of zebrafish embryos to bixafen caused severe retinal defects, including extreme microphthalmia and a significantly increased cell density of the ganglion cell layer (GCL). Compared with the controls, the expression levels of rod and cone photoreceptor marker genes (rho, opn1sw2, opn1mw1, opn1lw1, and opn1sw1) in the outer nuclear layer (ONL) were significantly downregulated after bixafen exposure. Furthermore, bixafen caused significantly increased expression levels in the GCL marker ath5 and decreased expression levels in the inner nuclear layer (INL) markers prox1a, vsx1, and sox2. Accordingly, we observed a significantly increased rate of cell apoptosis in the retina after bixafen exposure. Taken together, our data demonstrate that bixafen exhibits retinal developmental toxicity to zebrafish embryos/larvae, and thus, it may pose a significant environmental threat to aquatic organisms.

The Roles of Cullins E3 Ubiquitin Ligases in the Lipid Biosynthesis of the Green Microalgae Chlamydomonas reinhardtii.

Microalgae-based biodiesel production has many advantages over crude oil extraction and refinement, thus attracting more and more concern. Protein ubiquitination is a crucial mechanism in eukaryotes to regulate physiological responses and cell development, which is highly related to algal biodiesel production. Cullins as the molecular base of cullin-RING E3 ubiquitin ligases (CRLs), which are the largest known class of ubiquitin ligases, control the life activities of eukaryotic cells. Here, three cullins (CrCULs) in the green microalgae Chlamydomonas reinhardtii were identified and characterized. To investigate the roles of CrCULs in lipid metabolism, the gene expression profiles of CrCULs under nutrition starvation were examined. Except for down-regulation under nitrogen starvation, the CrCUL3 gene was induced by sulfur and iron starvation. CrCUL2 seemed insensitive to nitrogen and sulfur starvation because it only had changes after treatment for eight days. CrCUL4 exhibited an expression peak after nitrogen starvation for two days but this declined with time. All CrCULs expressions significantly increased under iron deficiency at two and four days but decreased thereafter. The silencing of CrCUL2 and CrCUL4 expression using RNAi (RNA interference) resulted in biomass decline and lipids increase but an increase of 20% and 28% in lipid content after growth for 10 days, respectively. In CrCUL2 and CrCUL4 RNAi lines, the content of fatty acids, especially C16:0 and C18:0, notably increased as well. However, the lipid content and fatty acids of the CrCUL3 RNAi strain slightly changed. Moreover, the subcellular localization of CrCUL4 showed a nuclear distribution pattern. These results suggest CrCUL2 and CrCUL4 are regulators for lipid accumulation in C. reinhardtii. This study may offer an important complement of lipid biosynthesis in microalgae.

Erchen Plus Huiyanzhuyu Decoction Inhibits the Growth of Laryngeal Carcinoma in a Mouse Model of Phlegm-Coagulation-Blood-Stasis Syndrome via the STAT3/Cyclin D1 Pathway.

Erchen plus Huiyanzhuyu decoction (EHD), a Chinese herbal medicine (CHM) formula that consists of Erchen decoction and Huiyanzhuyu decoction, has achieved satisfactory results in the clinic. The main function of EHD is to remove phlegm and blood stasis, and EHD is suitable for phlegm-coagulation-blood-stasis (PCBS) syndrome in laryngeal cancer (LC). In this study, a xenograft mouse model of LC with PCBS syndrome was constructed by feeding a high-fat diet, swimming in ice water, and subcutaneously injecting epinephrine hydrochloride for 2 weeks. Based on the successful Chinese medicine syndrome model, Hep2-luciferase-GFP cells were injected subcutaneously under the armpit of the right upper limb in mice to form tumours. A mouse model of LC with PCBS syndrome was established via heterotopic transplantation. Then, the mice received intragastric administration of different concentrations of EHD daily, and cisplatin (DDP) was intraperitoneally injected every week for 21 days. Tumour fluorescence in mice was measured with a living animal imager on days 7, 14, 21, and 28 during treatment. The results of this experiment confirmed that a mouse model of Chinese medicine syndrome was successfully constructed. Moreover, EHD slowed the growth of xenograft tumours in nude mice; decreased the expression levels of STAT3, p-STAT3, and cyclin D1; and upregulated the expression level of P27. In brief, EHD inhibited laryngeal tumour growth in a xenograft mouse model of PCBS syndrome and regulated the STAT3/cyclin D1 signalling pathway. This study was the first to construct a Chinese medicine xenograft mouse model of LC with PCBS syndrome; in addition, this study clarified that EHD regulated the STAT3/cyclin D1 signalling pathway to inhibit the growth of LC and that EHD may be a promising novel therapeutic compound for the treatment of patients with LC.

Erchen decoction plus huiyanzhuyu decoction inhibits the cell cycle, migration and invasion and induces the apoptosis of laryngeal squamous cell carcinoma cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Laryngeal carcinoma (LC) is one of the most common malignant head and neck cancers with high incidence and mortality rates. Erchen decoction plus Huiyanzhuyu decoction (EHD) is commonly used for treating LC patients and produces beneficial results. However, the mechanisms underlying the effects of EHD remain unclear. AIM OF THE STUDY: The present study aimed to analyse the anticancer effects of EHD on the LC cell cycle, apoptosis, migration and invasion in vitro and to explore the underlying biological mechanisms. MATERIALS AND METHODS: TU212 and Hep-2 cells were used. The antitumour effects of EHD were detected by CCK8, microscopy, flow cytometry, EdU incorporation, Hoechst 33342 staining, wound-healing, and transwell assays to assess viability, morphology, apoptosis, cell cycle, migration and invasion, respectively. Furthermore, STAT3 and related proteins were evaluated in laryngeal squamous cell carcinoma (LSCC) cells by Western blot (WB) analysis. RESULTS: EHD treatment significantly decreased STAT3 and p-STAT3 protein expression levels in LSCC cells. EHD blocked the cell cycle at the G0/G1 phase and induced LSCC apoptosis. Moreover, the viability, migration, and invasion of LSCC cells were markedly inhibited by EHD. In addition, the expression of the cell cycle-related proteins cyclin D1 and cyclin B1 was downregulated in LSCC cells, but P27 expression was increased after EHD treatment. Regarding apoptosis-related proteins, EHD also reduced Bcl-2 expression but upregulated Bax and caspase-3 expression in LSCC cells. In the migration- and invasion-related protein analyses, EHD downregulated MMP-9 expression and upregulated E-cadherin expression. CONCLUSIONS: These results suggest that EHD has an anticancer effect in LSCC. EHD treatment induces apoptosis and inhibits the cell cycle, migration and invasion of LSCC cells, but further work is warranted to address the mechanisms.