Effect of Xanthohumol from Humulus lupulus L. Against Gouty Bone Damage in Arthritis of Rats Induced by Mono-sodium Urate.

Xanthohumol (XAN) is an isoprenyl flavonoid from Humulus lupulus L. known for beer brewing, and an osteoprotective agent due to its active improvement in bone loss of osteoporosis. This study was first time to investigate its effects on anti-gouty bone injury in rats of gouty arthritis (GA) induced by monosodium urate (MSU). Results showed that XAN could significantly exert anti-inflammatory activity by alleviating swelling degree of joints, reducing serum level of inflammatory factors, improving inflammatory injury and degrading the Markin's score in lesion joint. Meanwhile, XAN could also fight against gouty bone damage by improving pathological changes of bone tissue and parameters of bone micro-structure. Moreover, XAN could even promote bone formation by effectively enhancing expression of Runx2 and OPG, while inhibit bone resorption with depressing matrix metalloproteinase-9 (MMP-9), MMP-13 and CTSK expression, reducing RANKL secretion, and abating the ratio of RANKL/OPG. Therefore, it was the first time to reveal the mechanism of XAN against gouty bone injury via inhibiting RANKL/OPG/RANK signaling pathway. Above all, this study provided potential strategy for the treatment of GA, and further contributed to research and resource development for hops.

Enhanced removal of ciprofloxacin and associated antibiotic-resistant genes from wastewater using a biological aeration filters in combination with Fe3O4-modified zeolite.

Antibiotics release into the water environment through sewage discharge is a significant environmental concern. In the present study, we investigated the removal of ciprofloxacin (CIP) in simulated sewage by biological aeration filter (BAF) equipped with Fe3O4-modified zeolite (Fe3O4@ZF). Fe3O4@ZF were prepared with impregnation method, and the Fe3O4 particles were successfully deposited on the surface of ZF in an amorphous form according to the results of XPS and XRD analysis. The modification also increased the specific surface area (from 16.22 m²/g to 22 m²/g) and pore volume (from 0.0047 cm³/g to 0.0063 cm³/g), improving the adsorption efficiency of antibiotics. Fe3O4 modified ZF improved the treatment performance significantly, and the removal efficiency of CIP in BAF-Fe3O4@ZF was 79%±2.4%. At 10ml/L CIP, the BAF-Fe3O4@ZF reduced the relative abundances of antibiotics resistance genes (ARGs) int, mexA, qnrB and qnrS in the effluent by 57.16%, 39.59%, 60.22%, and 20.25%, respectively, which effectively mitigate the dissemination risk of ARGs. The modification of ZF increased CIP-degrading bacteria abundance, such as Rhizobium and Deinococcus-Thermus, and doubled bacterial ATP activity, promoting CIP degradation. This study offers a viable, efficient method to enhance antibiotic treatment and prevent leakage via sewage discharge.

Crucial gating residues govern the enhancement of peroxygenase activity in an engineered cytochrome P450 O-demethylase.

P450-catalyzed O-demethylation reactions have recently attracted particular attention because of their potential applications in lignin bioconversion. We recently enabled the peroxygenase activity of CYP199A4, a NADH-dependent cytochrome P450 monooxygenase from Rhodopseudomonas palustris, by engineering a hydrogen peroxide (H2O2) tunnel. In this report, we reveal by crystallography and molecule dynamics simulations that key residues located at one of the water tunnels in CYP199A4 play a crucial gating role, which enhances the peroxygenase activity by regulating the inflow of H2O2. These results provide a more complete understanding of the mechanism by which monooxygenase is converted into peroxygenase activity through the H2O2 tunnel engineering (HTE) strategy. Furthermore, a library of engineered CYP199A4 peroxygenases was constructed to explore their application potentials for O-demethylation of various methoxy-substituted benzoic acid derivatives. The engineered CYP199A4 peroxygenases showed good functional group tolerance and preferential O-demethylation at the meta- or para-position, indicating potential O-demethylation of H- and G-type lignin monomers. This work reveals the feasibility of the HTE strategy in creating P450 peroxygenase from a mechanistic perspective, laying the foundation for developing an effective P450 O-demethylase applicable in lignin bioconversion.

Bajitianwan formula extract ameliorates bone loss induced by iron overload via activating RAGE/PI3K/AKT pathway based on network pharmacology and transcriptomic analysis.

Osteoporosis (OP) is closely related to iron overload. Bajitianwan (BJTW) is a traditional Chinese medicine formulation used for treating senile diseases such as dementia and osteoporosis. Modern pharmacological researches have found that BJTW has beneficial effect on bone loss and memory impairment in aging rats. This paper aimed to explore the role and mechanism of BJTW in ameliorating iron overload-induced bone loss. Furthermore, BJTW effectively improved the bone micro-structure of the femur in mice, and altered bone metabolism biomarkers alkaline phosphatase (ALP) and osteocalcin (OCN) in serum, as well as oxidative indexes superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) glutathione (GSH) and malondialdehyde (MDA) in liver. As for network pharmacology, 73 components collected from BJTW regulated 99 common targets merged in the BJTW and OP. The results of RNA-seq indicated that there were 418 potential targets in BJTW low dose group (BJTW-L) and 347 potential targets in BJTW high dose group (BJTW-H). Intriguingly, both PI3K-AKT signaling pathway and the AGEs-RAGE signaling pathway were contained in the KEGG pathways enrichment results of network pharmacology and transcriptomics, which were considered as the potential mechanism. Additionally, we verified that BJTW regulated the expression of related proteins in RAGE/PI3K-AKT pathways in MC3T3-E1 cells. In summary, BJTW has potent effect on protecting against iron overload-induced OP, and its mechanism may be related to the activation of the RAGE/PI3K-AKT signaling pathways.

BZW1 is a prognostic and immunological biomarker in pancreatic adenocarcinoma.

Pancreatic adenocarcinoma is the most common malignant tumor of the digestive system and is called the "king of cancer" because it has been labeled with high malignancy, rapid progression, poor survival, and poor prognosis. Previously, it was reported that the basic leucine zipper and W2 domains 1 (BZW1) is involved in the progression of many tumors. However, its research in digestive system tumors such as pancreatic cancer is rarely studied. To explore potential biomarkers related to survival and prognosis of pancreatic cancer and provide a new targeted therapy for it. We first analyzed the mRNA and protein expression of BZW1 in pancreatic cancer. We then explored the correlation of BZW1 with survival prognosis and immune infiltration in pancreatic cancer patients. Finally, we explored BZW1-related gene enrichment analysis, including protein-protein interaction networks, gene ontology functional enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. The mRNA and protein expression of the BZW1 gene in pancreatic cancer tissues were higher than those in adjacent normal tissues, and pancreatic cancer patients with high BZW1 expression had a poor prognosis. In addition, the expression of BZW1 was positively or negatively correlated with different immune cells of pancreatic cancer, such as CD4 + T lymphocytes, CD8 + T lymphocytes, B cells, macrophages, neutrophils, etc. Correlation enrichment analysis showed that we obtained 50 available experimentally determined BZW1-binding proteins and 100 targeted genes related to BZW1, and the intersection genes were eukaryotic translation termination factor 1 and Guanine nucleotide binding protein, alpha inhibiting activity polypeptide 3. Moreover, there was a positive correlation between BZW1 and eukaryotic translation termination factor 1 and Guanine nucleotide binding protein, alpha inhibiting activity polypeptide 3 genes in pancreatic cancer. Gene ontology enrichment analysis showed BZW1 was mainly related to biological processes such as "mRNA processing," "RNA splicing," "regulation of translational initiation," and "activation of innate immune response." The results of Kyoto Encyclopedia of Genes and Genomes pathway analysis further indicated that BZW1 may be involved in pancreatic carcinogenesis through the "spliceosome" and "ribosome." The BZW1 gene may be a potential immunotherapy target and a promising prognostic marker for pancreatic cancer.

Effects of repeated low-level red-light therapy on macular retinal thickness and microvascular system in children with myopia.

OBJECTIVE: The objective of the study was to use optical coherence tomography angiography (OCTA) to analyze the effects of repeated low-level red-light (LLLT) therapy on macular retinal thickness and the microvascular system in children with myopia to evaluate the safety of this therapy. METHODS: This prospective study included 40 school-age children with myopia (80 eyes), aged 7-14 years, who received therapy using a LLLT instrument. At baseline and therapy for 1 month, 3 months, 6 months, all children underwent comprehensive ophthalmological examinations, including slit-lamp examination, uncorrected visual acuity, best-corrected visual acuity, spherical equivalent degree, axial length, and OCTA. The vessel densities of the superficial retinal capillary plexus, macular inner retinal thickness, and full-layer retinal thickness were measured. RESULTS: The macular inner retinal thickness increased at 1 month and remained unchanged thereafter, It differed significantly in nine areas at 1, 3, and 6 months compared to the thicknesses before therapy (P < 0.05); however, we observed no significant differences between the different time points (P > 0.05). The macular full-layer retinal thickness increased at 1 month and remained unchanged thereafter; the changes showed significant differences at 1 month and 3 months compared to before therapy, for the inner nasal region (P < 0.05). The other eight areas showed significant differences at 1, 3, and 6 months compared with before therapy (P < 0.05); however, no significant difference was observed between the different time points after therapy (P > 0.05). The vessel density of the superficial retinal capillary plexus did not differ significantly among the four groups (P > 0.05). CONCLUSIONS: LLLT therapy was safe. The school-aged children exhibited macular thickening after LLLT therapy, which had no significant effect on macular microcirculation.

Gene cloning, protein expression, and enzymatic characterization of a double-stranded RNA degrading enzyme in Apolygus lucorum.

RNA interference (RNAi) is a powerful tool that post-transcriptionally silences target genes in eukaryotic cells. However, silencing efficacy varies greatly among different insect species. Recently, we met with little success when attempting to knock down genes in the mirid bug Apolygus lucorum via dsRNA injection. The disappearance of double-stranded RNA (dsRNA) could be a potential factor that restricts RNAi efficiency. Here, we found that dsRNA can be degraded in midgut fluids, and a dsRNase of A. lucorum (AldsRNase) was identified and characterized. Sequence alignment indicated that its 6 key amino acid residues and the Mg2+ -binding site were similar to those of other insects' dsRNases. The signal peptide and endonuclease non-specific domain shared high sequence identity with the brown-winged green stinkbug Plautia stali dsRNase. AldsRNase showed high salivary gland and midgut expression and was continuously expressed through the whole life cycle, with peaks at the 4th instar ecdysis in the whole body. The purified AldsRNase protein obtained by heterologously expressed can rapidly degrade dsRNA. When comparing the substrate specificity of AldsRNase, 3 specific substrates (dsRNA, small interfering RNA, and dsDNA) were all degraded, and the most efficient degradation is dsRNA. Subsequently, immunofluorescence revealed that AldsRNase was expressed in the cytoplasm of midgut cells. Through cloning and functional study of AldsRNase, the enzyme activity and substrate specificity of the recombinant protein, as well as the subcellular localization of nuclease, the reason for the disappearance of dsRNA was explained, which was useful in improving RNAi efficiency in A. lucorum and related species.

A review on ethnopharmacology, phytochemistry, pharmacology and potential uses of Portulaca oleracea L.

ETHNOPHARMACOLOGICAL RELEVANCE: Portulaca oleracea L. (PO), popularly known as purslane, has been documented in ethnopharmacology in various countries and regions. Traditional application records indicated that PO might be used extensively to treat the common cold, dysentery, urinary tract infections, coughing, eye infections, skin problems, gynecological diseases, and pediatric illnesses. AIM OF THE REVIEW: This paper includes a systematic review of the traditional usage, phytochemicals, pharmacological activity, and potential uses of PO to provide an overview of the research for further exploitation of PO resources. MATERIALS AND METHODS: This article uses "Portulaca oleracea L." and "purslane" as the keywords and collects relevant information on PO from different databases, including PubMed, Web of Science, Springer, Science Direct, ACS, Wiley, CNKI, Baidu Scholar, Google Scholar, and ancient meteria medica. RESULTS: PO is a member of the Portulacaceae family and is grown worldwide. Traditional Chinese medicine believes that purslane has the effect of improving eyesight, eliminating evil qi, quenching thirst, purgation, diuresis, hemostasis, regulating qi, promoting hair growth, detoxifying, and avoiding epidemic qi. Recent phytochemical investigations have shown that PO is a rich source of flavonoids, homoisoflavonoids, alkaloids, organic acids, esters, lignans, terpenoids, catecholamines, sterols, and cerebrosides. The purslane extracts or compounds have exhibited numerous biological activities such as anti-inflammatory, immunomodulatory, antimicrobial, antiviral, antioxidant, anticancer, renoprotective, hepatoprotective, gastroprotective, metabolic, muscle relaxant, anti-asthmatic and anti-osteoporosis properties. The significant omega-3 fatty acids, vital amino acids, minerals, and vitamins found in purslane also provide nutritional benefits. Purslane as a food/feed additive in the food industry and animal husbandry has caused concern. Its global wide distribution and tolerance to abiotic stress characteristics make it in the future sustainable development of agriculture a certain position. CONCLUSIONS: Based on traditional usage, phytochemicals, and pharmacological activity, PO is a potential medicinal and edible plant with diverse pharmacological effects. Due to purslane's various advantages, it may have vast application potential in the food and pharmaceutical industries and animal husbandry.

Visual impairment and blindness caused by retinal diseases: A nationwide register-based study.

Background: Retinal disorders cause substantial visual burden globally. Accurate estimates of the vision loss due to retinal diseases are pivotal to inform optimal eye health care planning and allocation of medical resources. The purpose of this study is to describe the proportion of visual impairment and blindness caused by major retinal diseases in China. Methods: A nationwide register-based study of vitreoretinal disease covering all 31 provinces (51 treating centres) of mainland China. A total of 28 320 adults diagnosed with retinal diseases were included. Participants underwent standardised ocular examinations, which included best-corrected visual acuity (BCVA), dilated-fundus assessments, and optical coherence tomography. Visual impairment and blindness are defined using BCVA according to the World Health Organization (WHO) (visual impairment: <20/63-≥20/400; blindness: <20/400) and the United States (visual impairment: <20/40-≥20/200; blindness: <20/200) definitions. The risk factors of vision loss were explored by logistic regression analyses. Results: Based on the WHO definitions, the proportions for unilateral visual impairment and blindness were 46% and 18%, respectively, whereas those for bilateral visual impairment and blindness were 31% and 3.3%, respectively. Diabetic retinopathy (DR) accounts for the largest proportion of patients with visual impairment (unilateral visual impairment: 32%, bilateral visual impairment: 60%) and blindness (unilateral blindness: 35%; bilateral blindness: 64%). Other retinal diseases that contributed significantly to vision loss included age-related macular degeneration, myopic maculopathy, retinal vein occlusion, and rhegmatogenous retinal detachment and other macular diseases. Women (bilateral vision loss: P = 0.011), aged patients (unilateral vision loss: 45-64 years: P < 0.001, ≥65 years: P < 0.001; bilateral vision loss: 45-64 years: P = 0.003, ≥65 years: P < 0.001 (reference: 18-44 years)) and those from Midwest China (unilateral and bilateral vision loss: both P < 0.001) were more likely to suffer from vision loss. Conclusions: Retinal disorders cause substantial visual burden among patients with retinal diseases in China. DR, the predominant retinal disease, is accountable for the most prevalent visual disabilities. Better control of diabetes and scaled-up screenings are warranted to prevent DR. Specific attention should be paid to women, aged patients, and less developed regions.

Role and Regulation of Transcription Factors in Osteoclastogenesis.

Bones serve mechanical and defensive functions, as well as regulating the balance of calcium ions and housing bone marrow.. The qualities of bones do not remain constant. Instead, they fluctuate throughout life, with functions increasing in some situations while deteriorating in others. The synchronization of osteoblast-mediated bone formation and osteoclast-mediated bone resorption is critical for maintaining bone mass and microstructure integrity in a steady state. This equilibrium, however, can be disrupted by a variety of bone pathologies. Excessive osteoclast differentiation can result in osteoporosis, Paget's disease, osteolytic bone metastases, and rheumatoid arthritis, all of which can adversely affect people's health. Osteoclast differentiation is regulated by transcription factors NFATc1, MITF, C/EBPα, PU.1, NF-κB, and c-Fos. The transcriptional activity of osteoclasts is largely influenced by developmental and environmental signals with the involvement of co-factors, RNAs, epigenetics, systemic factors, and the microenvironment. In this paper, we review these themes in regard to transcriptional regulation in osteoclastogenesis.

Traditional Chinese Medicine in Osteoporosis Intervention and the Related Regulatory Mechanism of Gut Microbiome.

The gut microbiome (GM) has become a crucial factor that can affect the progression of osteoporosis. A number of studies have demonstrated the impact of Traditional Chinese Medicine (TCM) on GM and bone metabolism. In this review, we summarize the potential mechanisms of the relationship between osteoporosis and GM disorder and introduce several natural Chinese medicines that exert anti-osteoporosis effects by modulating the GM. It is underlined that, through the provision of the microbial associated molecular pattern (MAMP), the GM causes inflammatory reactions and alterations in the Treg-Th17 balance and ultimately leads to changes in bone mass. Serotonin and many hormones, especially estrogen, may play a crucial role in the interaction of the GM with bone metabolism. Additionally, the GM may affect the absorption of specific nutrients in the intestine, particularly minerals like calcium, magnesium, and phosphorus. Several natural Chinese herbs, such as Sambucus Williamsii, Achyranthes bidentata Blume, Pleurotus ostreatus and Ganoderma lucidum mushrooms, Pueraria Lobata, and Agaricus blazei Murill have exhibited anti-osteoporosis effects through regulating the distribution and metabolism of the GM. These herbs may increase the abundance of Firmicutes, decrease the abundance of Bacteroides, promote the GM to produce more SCFAs, modulate the immune response caused by harmful bacteria, and increase the proportion of Treg-Th17 to indirectly affect bone metabolism. Moreover, gut-derived 5-HT is an important target for TCM to prevent osteoporosis via the gut-bone axis. Puerarin could prevent osteoporosis by improving intestinal mucosal integrity and decrease systemic inflammation caused by estrogen deficiency.

Anchoring a Structurally Editable Proximal Cofactor-like Module to Construct an Artificial Dual-center Peroxygenase.

A recent novel strategy for constructing artificial metalloenzymes (ArMs) that target new-to-nature functions uses dual-functional small molecules (DFSMs) with catalytic and anchoring groups for converting P450BM3 monooxygenase into a peroxygenase. However, this process requires excess DFSMs (1000 equivalent of P450) owing to their low binding affinity for P450, thus severely limiting its practical application. Herein, structural optimization of the DFSM-anchoring group considerably enhanced their binding affinity by three orders of magnitude (Kd ≈10-8  M), thus approximating native cofactors, such as FMN or FAD in flavoenzymes. An artificial cofactor-driven peroxygenase was thus constructed. The co-crystal structure of P450BM3 bound to a DFSM clearly revealed a precatalytic state in which the DFSM participates in H2 O2 activation, thus facilitating peroxygenase activity. Moreover, the increased binding affinity substantially decreases the DFSM load to as low as 2 equivalents of P450, while maintaining increased activity. Furthermore, replacement of catalytic groups showed disparate selectivity and activity for various substrates. This study provides an unprecedented approach for assembling ArMs by binding editable organic cofactors as a co-catalytic center, thereby increasing the catalytic promiscuity of P450 enzymes.

SeBLOS2 knockout via CRISPR/Cas9 leads to the loss of larval integument coloration in Spodoptera exigua (Lepidoptera: Noctuidae).

CRISPR/Cas9 technology is a precise and powerful tool for functionally exploring insect genes. The present study tested CRISPR/Cas9 as a way of undertaking effective gene mutagenesis in an important agricultural pest, the beet armyworm Spodoptera exigua. Based on a S. exigua transcriptome database, the entire complementary DNA sequence of SeBLOS2 encoding 140 amino acid residues was cloned. The gene was highly expressed in late larval stages (L3-L5). Using the CRISPR/Cas9 method, SeBLOS2 was knocked out by altering two sites in the coding region. This resulted in 70%-74% of the G0 generation (L4-L5) larvae displaying mosaic translucent integument. Four different mutations occurred at SeBLOS2-specific target sites, as demonstrated by further polymerase chain reaction-based genotypic analysis. Homozygote mutant L3 larvae were obtained in the G1 generation, with complete loss of white stripes and spots on their larval integument. These results demonstrate a crucial role of SeBLOS2 in integument pigmentation and suggest that the gene can act as a suitable nonlethal marker for functional research on genes in S. exigua and other Lepidopteran pests.

Enabling Peroxygenase Activity in Cytochrome P450 Monooxygenases by Engineering Hydrogen Peroxide Tunnels.

Given prominent physicochemical similarities between H2O2 and water, we report a new strategy for promoting the peroxygenase activity of P450 enzymes by engineering their water tunnels to facilitate H2O2 access to the heme center buried therein. Specifically, the H2O2-driven activities of two native NADH-dependent P450 enzymes (CYP199A4 and CYP153AM.aq) increase significantly (by >183-fold and >15-fold, respectively). Additionally, the amount of H2O2 required for an artificial P450 peroxygenase facilitated by a dual-functional small molecule to obtain the desired product is reduced by 95%-97.5% (with ∼95% coupling efficiency). Structural analysis suggests that mutating the residue at the bottleneck of the water tunnel may open a second pathway for H2O2 to flow to the heme center (in addition to the natural substrate tunnel). This study highlights a promising, generalizable strategy whereby P450 monooxygenases can be modified to adopt peroxygenase activity through H2O2 tunnel engineering, thus broadening the application scope of P450s in synthetic chemistry and synthetic biology.

Humulus lupulus L. Extract Protects against Senior Osteoporosis through Inhibiting Amyloid ß Deposition and Oxidative Stress in APP/PS1 Mutated Transgenic Mice and Osteoblasts

As aging progresses, ß-amyloid (Aß) deposition and the resulting oxidative damage are key causes of aging diseases such as senior osteoporosis (SOP). Humulus lupulus L. (hops) is an important medicinal plant widely used in the food, beverage and pharmaceutical industries due to its strong antioxidant ability. In this study, APP/PS1 mutated transgenic mice and Aß-injured osteoblasts were used to evaluate the protective effects of hops extracts (HLE) on SOP. Mice learning and memory levels were assessed by the Morris water maze. Mice femurs were prepared for bone micro-structures and immunohistochemistry experiments. The deposition of Aß in the hippocampus, cortex and femurs were determined by Congo red staining. Moreover, protein expressions related to antioxidant pathways were evaluated by Western blotting. It was found that HLE markedly improved learning abilities and ameliorated memory impairment of APP/PS1 mice, as well as regulated antioxidant enzymes and bone metabolism proteins in mice serum. Micro-CT tests indicated that HLE enhanced BMD and improved micro-architectural parameters of mice femur. More importantly, it was discovered that HLE significantly reduced Aß deposition both in the brain and femur. Further in vitro results showed HLE increased the bone mineralization nodule and reduced the ROS level of Aß-injured osteoblasts. Additionally, HLE increased the expression of antioxidant related proteins Nrf2, HO-1, NQO1, FoxO1 and SOD-2. These results indicated that Humulus lupulus L. extract could protect against senior osteoporosis through inhibiting Aß deposition and oxidative stress, which provides a reference for the clinical application of hops in the prevention and treatment of SOP.

Engineering Cytochrome P450BM3 Enzymes for Direct Nitration of Unsaturated Hydrocarbons.

Applications of the peroxidase activity of cytochrome P450 enzymes in synthetic chemistry remain largely unexplored. We present herein a protein engineering strategy to increase cytochrome P450BM3 peroxidase activity for the direct nitration of aromatic compounds and terminal aryl-substituted olefins in the presence of a dual-functional small molecule (DFSM). Site-directed mutations of key active-site residues allowed the efficient regulation of steric effects to limit substrate access and, thus, a significant decrease in monooxygenation activity and increase in peroxidase activity. Nitration of several phenol and aniline compounds also yielded ortho- and para-nitration products with moderate-to-high total turnover numbers. Besides direct aromatic nitration by P450 variants using nitrite as a nitrating agent, we also demonstrated the use of the DFSM-facilitated P450 peroxidase system for the nitration of the vinyl group of styrene and its derivatives.

Hybrid Machine Learning Approach for Evapotranspiration Estimation of Fruit Tree in Agricultural Cyber-Physical Systems.

The flourish of the Internet of Things (IoT) and data-driven techniques provide new ideas for enhancing agricultural production, where evapotranspiration estimation is a crucial issue in crop irrigation systems. However, tremendous and unsynchronized data from agricultural cyber-physical systems bring large computational costs as well as complicate performing conventional machine learning methods. To precisely estimate evapotranspiration with acceptable computational costs under the background of IoT, we combine time granulation computing techniques and gradient boosting decision tree (GBDT) with Bayesian optimization (BO) to propose a hybrid machine learning approach. In the combination, a fuzzy granulation method and a time calibration technique are introduced to break voluminous and unsynchronized data into small-scale and synchronized granules with high representativeness. Subsequently, GBDT is implemented to predict evapotranspiration, and BO is utilized to find the optimal hyperparameter values from the reduced granules. IoT data from Xi'an Fruit Technology Promotion Center in Shaanxi Province, China, verify that the proposed granular-GBDT-BO is effective for cherry tree evapotranspiration estimation with reduced computational time, and acceptable and robust predictive accuracy. Consequently, the precise estimation of crop evapotranspiration could provide operational guidance for plant irrigation, plant conservations, and pest control in the agricultural greenhouse.

TCM and Western Medicine: Diagnosis and Treatment of Novel Coronavirus Pneumonia, A Case Study.

Context: Clinical-practice observations have revealed that novel coronavirus pneumonia is related to the dampness pathogen; most patients show a systemic inflammatory response syndrome (SIRS), and patients with a severe form of the disease have sepsis. Objective: The current study aimed to explore the role of Traditional Chinese medicine (TCM) and western medicine in the treatment of COVID-19. Design: The research team developed a case study. Setting: The study took place at the Nanchang Ninth Hospital in Nanchang, China. Participant: The participant was a 44-year-old female patient with diabetes who was a hepatitis B carrier. She was admitted to the hospital and diagnosed with novel coronavirus pneumonia at admission. Intervention: She was treated for seven days with Western medicine, at which time the nucleic acid test for the virus was negative; however, she still had flaky glass shadows in the lungs. She was then treated with TCM. Results: The patient reached the discharge standard on February 29. A recheck was performed on March 6, six days after discharge. Her chest CT showed that the two lung lesions continued to be absorbed, and the viral nucleic acid test was negative. Conclusions: For diagnosis and treatment of novel coronavirus pneumonia, the determination of the use of six channels or Wei-qi-Ying-blood differentiation needs to be combined with syndrome differentiation and to comply with the body's process of eliminating pathogens. The recovery of qi, blood and body fluid, and SIRS should be taken into account.

Regiodivergent and Enantioselective Hydroxylation of C-H bonds by Synergistic Use of Protein Engineering and Exogenous Dual-Functional Small Molecules.

It is a great challenge to optionally access diverse hydroxylation products from a given substrate bearing multiple reaction sites of sp3 and sp2 C-H bonds. Herein, we report the highly selective divergent hydroxylation of alkylbenzenes by an engineered P450 peroxygenase driven by a dual-functional small molecule (DFSM). Using combinations of various P450BM3 variants with DFSMs enabled access to more than half of all possible hydroxylated products from each substrate with excellent regioselectivity (up to >99 %), enantioselectivity (up to >99 % ee), and high total turnover numbers (up to 80963). Crystal structure analysis, molecular dynamic simulations, and theoretical calculations revealed that synergistic effects between exogenous DFSMs and the protein environment controlled regio- and enantioselectivity. This work has implications for exogenous-molecule-modulated enzymatic regiodivergent and enantioselective hydroxylation with potential applications in synthetic chemistry.

CRISPR/Cas9-mediated methoprene-tolerant 1 knockout results in precocious metamorphosis of beet armyworm (Spodoptera exigua) only at the late larval stage.

Juvenile hormone (JH) controls almost every aspect of an insect, especially metamorphosis. Since RNA interference works on transcripts and is often insufficient in Lepidoptera, how JH affects larval development in these insects is not well studied. Using the CRISPR/Cas9 technique, we knocked out Spodoptera exigua methoprene-tolerant 1 (SeMet1) gene of beet armyworm by modifying two sites in the coding region. However, SeMet1 knockout did not affect egg hatch rate or larval development at L1-L3 stages. In contrast to the consistent five larval instars of the control group, L4 SeMet1 mutants began to show signs of precocious metamorphosis, that is, small patches of pupal cuticle. Most L4 and all L5 SeMet1 mutants died for failing to shed their mosaic cuticles. RNA-seq indicated that most genes encoding pupal cuticle proteins and chitinase genes were altered in SeMet1 mutant L4 larvae. SeKr-h1, a key transcription factor in JH action was significantly down-regulated in L3-L5 larvae, while SeBR-C, a pupal indicator was only upregulated in L4-L5 larvae. These results suggested that S. exigua larvae may initially develop independently of JH, and involve SeMet1 in transducing JH signalling, leading to controlled larval metamorphosis at the late larval stage. We believe our findings will enhance better understanding of JH regulation of larval development.