"Mini-community" simulation revealed the differences of endophytic fungal communities between the above- and below-ground tissues of Ephedra sinica Stapf.

The microecology of endophytic fungi in special habitats, such as the interior of different tissues from a medicinal plant, and its effects on the formation of metabolites with different biological activities are of great importance. However, the factors affecting fungal community formation are unclear. This study is the first to utilize "mini-community" remodeling to understand the above phenomena. First, high-throughput sequencing technology was applied to explore the community composition and diversity of endophytic fungi in the above-ground tissues (Ea) and below-ground tissues (Eb) of Ephedra sinica. Second, fungi were obtained through culture-dependent technology and used for "mini-community" remodeling in vitro. Then, the effects of environmental factors, partner fungi, and plant tissue fluid (internal environment) on endophytic fungal community formation were discussed. Results showed that environmental factors played a decisive role in the selection of endophytic fungi, that is, in Ea and Eb, 93.8% and 25.3% of endophytic fungi were halophilic, respectively, and 10.6% and 60.2% fungi were sensitive to high temperature (33 °C), respectively. Meanwhile, pH had little effect on fungal communities. The internal environment of the plant host further promoted the formation of endophytic fungal communities.

Andrographolide causes p53-independent HCC cell death through p62 accumulation and impaired DNA damage repair.

BACKGROUND: Hepatocellular carcinoma (HCC) is a highly lethal cancer characterized by dominant driver mutations, including p53. Consequently, there is an urgent need to search for novel therapeutic agents to treat HCC. Andrographolide (Andro), a clinically available anti-inflammatory phytochemical agent, has shown inhibitory effects against various types of cancer, including HCC. However, the underlying molecular mechanisms of its action remain poorly understood. PURPOSE: This study aims to investigate the molecular mechanisms by which p53 and p62 collectively affect Andro-induced HCC cell death, using both in vitro and in vivo models. METHODS: In vitro cellular experiments were conducted to examine the effects of Andro on cell viability and elucidate its mechanisms of action. In vivo xenograft experiments further validated the anti-cancer effects of Andro. RESULTS: Andro induced dose- and time-dependent HCC cell death while sparing normal HL-7702 hepatocytes. Furthermore, Andro caused DNA damage through the generation of reactive oxygen species (ROS), a critical event leading to cell death. Notably, HCC cells expressing p53 exhibited greater resistance to Andro-induced cell death compared to p53-deficient cells, likely due to the ability of p53 to induce G2/M cell cycle arrest. Additionally, Andro-induced p62 aggregation led to the proteasomal degradation of RAD51 and 53BP1, two key proteins involved in DNA damage repair. Consequently, silencing or knocking out p62 facilitated DNA damage repair and protected HCC cells. Importantly, disruption of either p53 or p62 did not affect the expression of the other protein. These findings were further supported by the observation that xenograft tumors formed by p62-knockout HCC cells displayed increased resistance to Andro treatment. CONCLUSION: This study elucidates the mechanistic basis of Andro-induced HCC cell death. It provides valuable insights for repurposing Andro for the treatment of HCC, regardless of the presence of functional p53.

Bibliometric analysis of acupuncture for headache from 1974 to 2022: A scoping literature review based on international database.

This study aimed to investigate the research hotspots and global trends of acupuncture in the treatment of headaches from 1974 to 2022. The Web of Science core collection database and literature related to acupuncture for headache treatment were retrieved. The CiteSpace (version 5.1.R8) and VOSviewer (version 1.6.19) software perform collaborative network analysis on the information of countries, academic institutions, authors, and co-occurrence network analysis on keywords, co-cited journals, and references. A total of 841 studies were included. Overall, the number of publications has increased over the past 5 decades. We identified and analyzed the countries, institutions, authors, and journals that were most active in the domain of acupuncture treatment for headaches. The most productive countries were the United States and China. Chengdu University of Traditional Chinese Medicine was the most productive institution and Linde Klaus was the most productive author. Cephalalgia was the most productive and co-cited journal, whereas Lancet had the highest impact factor. The research hotspots mainly focus on headache, migraine, tension headache, electroacupuncture, and acupuncture. Research trends have mainly focused on acupuncture therapy and its curative effects, migraine without aura, paroxysmal migraine, and the mechanism of acupuncture treatment. The main research hotspots and frontier trends were the therapeutic effect and mechanism of acupuncture for headaches. The mechanism of acupuncture in the treatment of headache mainly focused on the neural mechanism by multimodal MRI.

HIF2α activation and mitochondrial deficit due to iron chelation cause retinal atrophy.

Iron accumulation causes cell death and disrupts tissue functions, which necessitates chelation therapy to reduce iron overload. However, clinical utilization of deferoxamine (DFO), an iron chelator, has been documented to give rise to systemic adverse effects, including ocular toxicity. This study provided the pathogenic and molecular basis for DFO-related retinopathy and identified retinal pigment epithelium (RPE) as the target tissue in DFO-related retinopathy. Our modeling demonstrated the susceptibility of RPE to DFO compared with the neuroretina. Intriguingly, we established upregulation of hypoxia inducible factor (HIF) 2α and mitochondrial deficit as the most prominent pathogenesis underlying the RPE atrophy. Moreover, suppressing hyperactivity of HIF2α and preserving mitochondrial dysfunction by α-ketoglutarate (AKG) protects the RPE against lesions both in vitro and in vivo. This supported our observation that AKG supplementation alleviates visual impairment in a patient undergoing DFO-chelation therapy. Overall, our study established a significant role of iron deficiency in initiating DFO-related RPE atrophy. Inhibiting HIF2α and rescuing mitochondrial function by AKG protect RPE cells and can potentially ameliorate patients' visual function.

Study on the Effects of Different Doses of Dahuang Zhechong Pills on the Ubiquitin Proteasome Pathway/Nuclear Factor-κB in Rats with Atherosclerosis and Its Mechanism.

In order to investigate the effects of different doses of Dahuang Zhechong pills on the ubiquitin proteasome pathway/nuclear factor-κB (UPP-NF-κB) in rats with atherosclerosis (AS), 58-week-old male Wistar rats were selected and randomly divided into the normal group, model group, control group, low-dose group, and high-dose group. The model group and the drug group are given intraperitoneal injections of vitamins, and the model group and the drug group are given a high-fat diet. Rats in the low-dose group and high-dose group are given low-dose and high-dose Dahuang Zhechong pill lavage solution, respectively. Besides, the control group is given simvastatin solution by gavage, and intervention is performed once a day for 12 weeks. Ubiquitin (Ub) protein expression, ubiquitin activase (UBE1), nuclear factor-κB, nuclear inhibitory factor-κB (IκB) gene expression, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and serum tumor necrosis factor-α (TNF-α) are compared. The experimental result shows that Dahuang Zhechong pills can reduce inflammation and prevent and treat AS by blocking the activation of the UPP/NF-κB signaling pathway and can be used as a proteasome inhibitor in the clinical treatment of AS.

Long-term vitamin A supplementation in a preclinical mouse model for RhoD190N-associated retinitis pigmentosa.

Retinitis pigmentosa (RP) is caused by one of many possible gene mutations. The National Institutes of Health recommends high daily doses of vitamin A palmitate for RP patients. There is a critical knowledge gap surrounding the therapeutic applicability of vitamin A to patients with the different subtypes of the disease. Here, we present a case report of a patient with RP caused by a p.D190N mutation in Rhodopsin (RHO) associated with abnormally high quantitative autofluorescence values after long-term vitamin A supplementation. We investigated the effects of vitamin A treatment strategy on RP caused by the p.D190N mutation in RHO by exposing Rhodopsin p.D190N (RhoD190N/+) and wild-type (WT) mice to experimental vitamin A-supplemented and standard control diets. The patient's case suggests that the vitamin A treatment strategy should be further studied to determine its effect on RP caused by p.D190N mutation in RHO and other mutations. Our mouse experiments revealed that RhoD190N/+ mice on the vitamin A diet exhibited higher levels of autofluorescence and lipofuscin metabolites compared to WT mice on the same diet and isogenic controls on the standard control diet. Vitamin A supplementation diminished photoreceptor function in RhoD190N/+ mice while preserving cone response in WT mice. Our findings highlight the importance of more investigations into the efficacy of clinical treatments like vitamin A for patients with certain genetic subtypes of disease and of genotyping in the precision care of inherited retinal degenerations.

Residue dissipation, transfer and safety evaluation of picoxystrobin during tea growing and brewing.

BACKGROUND: Picoxystrobin is a new osmotic and systemic broad-spectrum methoxyacrylate fungicide with a good control effect on tea anthracnose, so it has been proposed to spray picoxystrobin before the occurrence and onset of tea anthracnose during tea bud growth in order to protect them. However, there are few reports about the residue analysis method, field dissipation, terminal residue and risk assessment of picoxystrobin in tea. And there is no scientific and reasonable maximum residue limit of picoxystrobin in green tea. RESULTS: A rapid and sensitive analysis method for picoxystrobin residue in fresh tea leaf, green tea, tea infusion and soil was established by UPLC-MS/MS. The spiked recoveries of picoxystrobin ranged from 73.1% to 111.0%, with relative standard deviations from 1.8% to 9.2%. The limits of quantitation were 20 µg kg-1 in green tea, 8 µg kg-1 in fresh tea leaves and soil and 0.16 µg kg-1 in tea infusion. The dissipation half-lives of picoxystrobin in fresh tea leaf and soil were 2.7-6.8 and 2.5-14.4 days, respectively. And the maximum residue of picoxystrobin in green tea was 15.28 mg kg-1 with PHI at 10 days for terminal test. The total leaching rate of picoxystrobin during green tea brewing was lower than 35.8%. CONCLUSIONS: According to safety evaluation, the RQc and RQa values of picoxystrobin in tea after 5 to 14 days for the last application were significantly lower than 1. Therefore, the maximum residue limit value of picoxystrobin in tea that we suggest to set at 20 mg kg-1 can ensure the safety of tea for human drinking. © 2020 Society of Chemical Industry.

Application and enantioselective residue determination of chiral pesticide penconazole in grape, tea, aquatic vegetables and soil by ultra performance liquid chromatography-tandem mass spectrometry.

Penconazole is a typical triazole fungicide with wide use on fruits, vegetables, and tea plants to control powdery mildew. In the present study, an efficient graphite carbon black solid phase extraction (GCB-SPE) purification combined with chiral ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed for determination of penconazole enantiomers in different complex matrices, including grape, tea, soil, lotus root, lotus leaf, lotus seed and hulls. The method was then applied to investigate the enantioselective dissipation of penconazole enantiomers in a real field experiment of grape and soil. As a result, a satisfactory separation of penconazole enantiomers on a chiral Lux Cellulose-2 column (150 mm × 2 mm i.d., 3 µm) was obtained with 0.1% formic acid in methanol and 10 mmol L-1 ammonium acetate in water (75/25, v/v) as mobile phase at 0.25 mL min-1. The enantiomer (+)-penconazole was firstly eluted, and (-)-penconazole was then eluted. The method showed reliable performances in linearity, recovery and precision, the recoveries of (+)-penconazole and (-)-penconazole in all of six matrices were between 70.5% and 121.0% with the relative standard deviations (RSDs) ranging from 0.8% to 23.6% at the low, medium and high spiked levels. The limits of quantitation (LOQs) of this method were lower than 0.0025 mg kg-1 in grape, soil and lotus root, 0.005 mg kg-1 in lotus leaf, lotus seed meat and lotus seed shell, and 0.0125 mg kg-1 in tea. Results of field trials indicated that (-)-penconazole degraded faster than its (+)-isomer in grape. While only a moderate stereoselectivity was observed in soil, with (-)-penconazole preferential degraded. The proposed method could be used to investigate enantioselective environmental behavior of penconazole enantiomers in complex matrices. And results in this study could provide useful information on realistic risk assessment of penconazole in grape.

Multimodal characterization of a novel mutation causing vitamin B6-responsive gyrate atrophy.

PURPOSE: Gyrate atrophy (GA) is a rare chorioretinal degeneration that results in the deterioration of night and peripheral vision, eventually leading to blindness. The disorder is caused by mutations in the gene encoding ornithine aminotransferase (OAT), causing increased levels of plasma ornithine. Treatment revolves around lowering plasma ornithine levels, with vitamin B6 supplementation being the preferred treatment. Nevertheless, most patients do not respond to this therapy. Here, we report a rare case of vitamin B6-responsive GA caused by a novel mutation in OAT and characterize the presentation with multimodal imaging. METHODS: This is a single-patient case report with a clinical diagnosis based on history, multimodal retinal imaging, laboratory findings, and DNA sequencing analysis. We include a 3D structure prediction of the novel mutant protein. RESULTS: DNA sequencing analysis demonstrated that there is a homozygous, novel variant c.473A>C: p.Y158S in OAT. Upon undergoing two weeks of vitamin B6 supplementation, the patient exhibited a 28.5% reduction in plasma ornithine levels. In a follow-up visit two years later, plasma ornithine levels were reduced by 24.1% from the levels at initial presentation and disease progression was retarded based on clinical findings. CONCLUSION: One novel homozygous missense mutation in OAT was identified and considered to be pathogenic in a patient with GA. The response for the vitamin B6 supplementation was positive, which is rare in all the GA cases reported in the literature. Our data suggests that further studies regarding the relationship between genotype and responsiveness to vitamin B6 should be conducted.