Retron-mediated multiplex genome editing and continuous evolution in Escherichia coli.

While there are several genome editing techniques available, few are suitable for dynamic and simultaneous mutagenesis of arbitrary targeted sequences in prokaryotes. Here, to address these limitations, we present a versatile and multiplex retron-mediated genome editing system (REGES). First, through systematic optimization of REGES, we achieve efficiency of ∼100%, 85 ± 3%, 69 ± 14% and 25 ± 14% for single-, double-, triple- and quadruple-locus genome editing, respectively. In addition, we employ REGES to generate pooled and barcoded variant libraries with degenerate RBS sequences to fine-tune the expression level of endogenous and exogenous genes, such as transcriptional factors to improve ethanol tolerance and biotin biosynthesis. Finally, we demonstrate REGES-mediated continuous in vivo protein evolution, by combining retron, polymerase-mediated base editing and error-prone transcription. By these case studies, we demonstrate REGES as a powerful multiplex genome editing and continuous evolution tool with broad applications in synthetic biology and metabolic engineering.

One-Step Pyrolysis Construction of Bimetallic Atom-Cluster Sites for Boosting Bifunctional Catalytic Activity in Zn-Air Batteries.

Due to their unique advantages, single atoms and clusters of transition metals are expected to achieve a breakthrough in catalytic activity, but large-scale production of active materials remains a challenge. In this work, a simple solvent-free one-step annealing method is developed and applied to construct diatomic and cluster active sites in activated carbon by utilizing the strong anchoring ability of phenanthroline to metal ions, which can be scaled for mass productions. Benefiting from the synergy between the different metals, the obtained sub-nano-bimetallic atom-cluster catalysts (FeNiAC -NC) exhibit high oxygen reduction reactions (ORR) activity (E1/2 = 0.936 V vs. RHE) and a small ORR/oxygen evolution reaction (OER) potential gap of only 0.594 V. An in-house pouch Zn-air battery is assembled using an FeNiAC -NC catalyst, which demonstrates a stability of 1000 h, outperforming previous reports. The existence of clusters and their effects on catalytic activity is analyzed by density functional theory calculations to reveal the chemistry of nano-bimetallic atom-cluster catalysts.

Efficient production of l-glutathione by whole-cell catalysis with ATP regeneration from adenosine.

l-glutathione (GSH) is an important tripeptide compound with extensive applications in medicine, food additives, and cosmetics industries. In this work, an innovative whole-cell catalytic strategy was developed to enhance GSH production by combining metabolic engineering of GSH biosynthetic pathways with an adenosine-based adenosine triphosphate (ATP) regeneration system in Escherichia coli. Concretely, to enhance GSH production in E. coli, several genes associated with GSH and  l-cysteine degradation, as well as the branched metabolic flow, were deleted. Additionally, the GSH bifunctional synthase (GshFSA) and GSH ATP-binding cassette exporter (CydDC) were overexpressed. Moreover, an adenosine-based ATP regeneration system was first introduced into E. coli to enhance GSH biosynthesis without exogenous ATP additions. Through the optimization of whole-cell catalytic conditions, the engineered strain GSH17-FDC achieved an impressive GSH titer of 24.19 g/L only after 2 h reaction, with a nearly 100% (98.39%) conversion rate from the added  l-Cys. This work not only unveils a new platform for GSH production but also provides valuable insights for the production of other high-value metabolites that rely on ATP consumption.

Gas-Solid Reaction Synthesis and Electrocatalytic Oxygen Reduction of Pt-Skin L10-PtFe/C.

Compared to Pt/C, the atomic ordered Pt-based intermetallic compounds can deliver higher efficiency and reliable stability, and they are considered one of the ideal cathode catalysts for the next generation of fuel cells. This work proposed a simple ferrocene atmosphere annealing method to improve commercial Pt/C and convert Pt to L10-PtFe. After further acid etching treatment, the obtained carbon-supported Pt-skin L10-PtFe (Pt-skin L10-PtFe/C) with superfine particle size (∼3.3 nm) not only was highly dispersed on the carbon but possesses a thin Pt skin, like the armor of L10-PtFe. As excepted, the ORR activity of Pt-skin L10-PtFe/C (0.375 A mg-1; 0.921 mA cm-2) is far better than that of commercial Pt/C (0.121 A mg-1; 0.260 mA cm-2), and its stability is also greatly improved. Our proposed gas-solid reaction is straightforward and has great potential in producing Pt-based intermetallic catalysts on a large scale.

Development of an UHPLC-MS/MS method to determine cutaneous biodistribution of cannabidiol after topical application of cannabidiol gel assisted by iontophoresis.

Cannabidiol has potential for use in skin disease therapy, so it is important to know the cutaneous biodistribution of cannabidiol after topical application of cannabidiol formulations. However, currently existing quantification methods for the investigation of cannabidiol skin distribution are not optimal. This study aimed to establish a method for the determination of cannabidiol in skin samples by UHPLC-MS/MS. A BEH C18 (50.0 × 2.1 mm, 2.5 µm) column was used; the mobile phase consisted of acetonitrile-0.1% formic acid (70:30, v/v), the flow rate was 0.2 µl·min-1 and the column temperature was 30°C. Positive-ion mode with multiple reaction monitoring detection was used to quantify cannabidiol (m/z 315.1 → 193.1) while diphenhydramine (m/z 256.3 → 167.08) served as the internal standard. Good linearity was shown in the range of 1-200 ng·ml-1 for cannabidiol with correlation coefficients of >0.999. The LLOQ was 1 ng·ml-1 . The intra-day and inter-day RSDs of cannabidiol were all <2%. A cryo-sectioning technique combined with the UHPLC-MS/MS method was used to successfully determine cannabidiol levels in a series of very thin skin layers.

Efficacy comparison between iliosacral screw fixation of the posterior pelvic ring fracture with the assistance of modified percutaneous three-dimensional printing guide template and conventional fluoroscopy. / æ”¹è‰¯ç»çš®ä¸‰ç»´æ‰“å°å¯¼æ¿ä¸Žä¼ ç»Ÿé€è§†è¾ åŠ©éª¶é«‚èžºé’‰å›ºå®šéª¨ç›†åŽçŽ¯éª¨æŠ˜çš„ç–—æ•ˆæ¯”è¾ƒ.

OBJECTIVES: The effect of three-dimensional (3D) printed bone-attached guide plate assisted cannulated screw fixation of pelvic fracture is reliable, but extensive soft tissue dissection is still required when installing the guide plate. This study aims to compare the efficacy of posterior pelvic ring fracture fixation with iliosacral screw insertion between the assistance of modified percutaneous patient specific 3D printed guide template and conventional fluoroscopy. METHODS: From May, 2019 and September 2021, 28 patients sustained posterior pelvic ring fractures were randomized into 2 groups: A guide template group, in which the iliosacral screw was inserted for fixation of the posterior pelvic ring fracture with the assistance of modified percutaneous patient specific 3D printed guide template, and a fluoroscopy group, in which the iliosacral screw was inserted under the guidance of conventional fluoroscopy. The operation time, fluoroscopic frequency, intraoperative blood loss, and incision length were recorded for each screw insertion. Fracture reduction was evaluated according to the Matta criteria. The screw position was evaluated according to the modified Gras classification, and the functional outcome was evaluated according to Majeed score. The parameters of both groups were compared, and statistical analysis was performed. RESULTS: All the 28 patients were followed up for 12-24 months. Of them, 15 iliosacral screws were inserted in 14 patients in the guide template group, and 14 iliosacral screws were inserted in 14 patients in the fluoroscopy group. The operation time, fluoroscopic frequency, screw deviation, incision length, and blood loss in the guide template group were 20-30(25.8±2.8) min, 9-15(12.2±1.9), 2-4(2.6±0.7) mm, 4-5(4.6±0.5) cm, and 5-10 (7.8±1.7) mL, respectively, whereas those in the fluoroscopy group were 30-60(48.1±7.5) min, 40-96(64.7±16.3), 3-6(4.2±0.9) mm, 0.8-1.2(1.0±0.1) cm, and 2-5(3.1±1.3) mL, respectively, and there were statistical significance (all P<0.001). Fracture reduction was evaluated according to the Matta criteria, and all the patients reached excellence and good (P=0.584) in the 2 groups. According to modified Gras classification, there were 12 Grade I screws, 3 Grade II screws, and 0 Grade III screws in the guide template group, and 10 Grade I screws, 3 Grade II screws, and 1 Grade III screw in the fluoroscopy group, with no statistical significance (P=0.334). The functional outcome was evaluated according to Majeed score at the last follow-up, without significant difference between the guide template group and the fluoroscopy group (P=0.908). CONCLUSIONS: Compared with the conventional fluoroscopy, it would cost less operation time, less fluoroscopic frequency and increase more accurate screw insertion to fixate the posterior pelvic ring fracture with the assistance of modified percutaneous patient specific 3D printed guide template.

Salidroside Inhibits Reactive Astrogliosis and Glial Scar Formation in Late Cerebral Ischemia via the Akt/GSK-3ß Pathway.

Cerebral ischemia leads to reactive astrogliosis and glial scar formation. Glial scarring can impede functional restoration during the recovery phase of stroke. Salidroside has been shown to have neuroprotective effects after ischemic stroke, but its impact on long-term neurological recovery, especially whether it regulates reactive astrogliosis and glial scar formation, is unclear. In this study, male adult C57/BL6 mice were subjected to transient cerebral ischemia injury followed by intravenous salidroside treatment. Primary astrocytes were treated with lipopolysaccharide (LPS) or conditioned medium from cultured primary neurons subjected to oxygen-glucose deprivation (CM-OGD). Salidroside significantly improved long-term functional outcomes following ischemic stroke in the rotarod and corner tests. It also reduced brain glial scar volume and decreased expression of the glial scar marker, glial fibrillary acidic protein (GFAP) and inhibited astrocyte proliferation. In primary astrocyte cultures, salidroside protected astrocytes from CM-OGD injury-induced reactive astroglial proliferation, increasing the percentage of cells in G0/G1 phase and reducing the S populations. The inhibitory effect of salidroside on the cell cycle was related to downregulation of cyclin D1 and cyclin-dependent kinase 4 (CDK4) mRNA expression and increased p27Kip1 mRNA expression. Similar results were found in the LPS-stimulated injury model in astroglial cultures. Western blot analysis demonstrated that salidroside attenuated the CM-OGD-induced upregulation of phosphorylated Akt and glycogen synthase kinase 3ß (GSK-3ß). Taken together, these results suggested that salidroside can inhibit reactive astrocyte proliferation, ameliorate glial scar formation and improve long-term recovery, probably through its effects on the Akt/GSK-3ß pathway.

Identification of Long-Distance Transmissible mRNA between Scion and Rootstock in Cucurbit Seedling Heterografts.

: Grafting has been widely used to improve plant growth and tolerance in crop production, as well as for clarifying systemic mRNA signaling from donor to recipient tissues in organ-to-organ communication. In this study, we investigated graft partner interaction mechanisms of Cucumis sativus (Csa) and Cucurbita moschata (Cmo) using a large-scale endogenous mRNA transport. The results indicated that most mobile transcripts followed an allocation pathway from source to sink. Gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that mRNA mobility functions are universally common and individually specific. Identification of mRNA mobility between distant tissues in heterografts with RT-PCR (reverse transcription PCR), RT-qPCR (reverse transcriptional quantitative real time PCR), and clone sequencing were used to estimate 78.75% of selected mobile transcripts. Integration of bioinformatic analysis and RT-qPCR identification allowed us to hypothesize a scion-to-rootstock-to-scion feedback signal loop of Csa move-down and Cmo move-up mRNAs, where Csa scion move-down mRNAs were involved in carbon fixation and biosynthesis of amino acid pathways, and Cmo root received Csa move-down mRNA and then delivered the corresponding Cmo upward mRNA to scion to improve photosynthesis of cucumber scion. This formed a feedback signal loop of scion-to-rootstock-to scion to explain why pumpkin rootstock enhanced cucumber production in the industry, which was utilized for organ communication and mediates photosynthesis processes in heterograft cucurbit crops.

Genome-Wide Characterization of GRAS Family and Their Potential Roles in Cold Tolerance of Cucumber (Cucumis sativus L.).

Cucumber (Cucumis sativus L.) is one of the most important cucurbit vegetables but is often subjected to stress during cultivation. GRAS (gibberellic acid insensitive, repressor of GAI, and scarecrow) genes encode a family of transcriptional factors that regulate plant growth and development. In the model plant Arabidopsis thaliana, GRAS family genes function in formation of axillary meristem and root radial structure, phytohormone (gibberellin) signal transduction, light signal transduction and abiotic/biological stress. In this study, a gene family was comprehensively analyzed from the aspects of evolutionary tree, gene structure, chromosome location, evolutionary and expression pattern by means of bioinformatics; 37 GRAS gene family members have been screened from cucumber. We reconstructed an evolutionary tree based on multiple sequence alignment of the typical GRAS domain and conserved motif sequences with those of other species (A. thaliana and Solanum lycopersicum). Cucumber GRAS family was divided into 10 groups according to the classification of Arabidopsis and tomato genes. We conclude that tandem and segmental duplication have played important roles in the expansion and evolution of the cucumber GRAS (CsaGRAS) family. Expression patterns of CsaGRAS genes in different tissues and under cold treatment, combined with gene ontology annotation and interaction network analysis, revealed potentially different functions for CsaGRAS genes in response to cold tolerance, with members of the SHR, SCR and DELLA subfamilies likely playing important roles. In conclusion, this study provides valuable information and candidate genes for improving cucumber tolerance to cold stress.

Chemical Reaction-Driven Spreading of an Organic Extractant on the Gas-Water Interface: Insight into the Controllable Formation of a Gas Bubble-Supported Organic Extractant Liquid Membrane.

The extraction and recovery of low-concentration valuable metals from various complex aqueous solutions or industrial waste waters have attracted extensive interests in recent years. In our previous works, we suggested a novel technique called bubbling organic liquid membrane extraction by spreading and covering an organic extractant with extremely small volume on the surface of gas bubbles to form a layer of the gas bubble-supported organic liquid membrane for selective extraction and enrichment of low-concentration targets from dilute aqueous solutions. It was found that for successfully performing the bubbling organic liquid membrane extraction, a prerequisite is knowing how to control the formation of a stable organic liquid membrane covered on the surface of gas bubbles. However, once the organic extractant starts to spread on the surface of gas bubbles, the extraction chemical reaction at the interface between the organic extractant liquid membrane and the rare-earth aqueous solution will occur. In the present work, the spreading behavior of the organic extractant P507 on the surface of rare-earth aqueous solutions was investigated and was compared with the behaviors on the surface of deionized water. It was revealed that the spreading of the organic extractant P507 on the surface of aqueous solutions containing rare-earth ions was accelerated because of the occurrence of the chemical reactions at the gas-water interface. The difference in the spreading rate of organic extractant P507 liquid droplets on the surface of deionized water and on that of Er(III) aqueous solutions with an increase in the P507 concentration, the saponification degrees of the P507 extractant, and the preloading amount of Er(III) in the P507 extractant revealed that the chemical reaction at the interface between the spreading P507 thin liquid membrane and the Er(III) aqueous solution would result in the Marangoni convection along the interface, which is in favor of overcoming the resistance from the viscous force when the surface tension gradient replaces gravity as a dominant driving force for the spreading. The present work provides an experimental foundation toward understanding the effect of the interfacial chemical reaction on the spreading behavior of an organic oil droplet on the gas-water interface. It is beneficial for the development of our suggested new technique of bubbling organic liquid membrane extraction and to achieve a controllable generation of a stable gas bubble-supported organic liquid membrane for performing solvent extraction at large aqueous-to-oil phase ratios.

Metabolic pathway engineering for high-level production of 5-hydroxytryptophan in Escherichia coli.

Cellular metabolic networks should be carefully balanced using metabolic engineering to produce the desired products at the industrial scale. As the precursor for the biosynthesis of the neurotransmitter serotonin, 5-hydroxytryptophan (5-HTP) is effective in treating a variety of diseases, such as depression, fibromyalgia, obesity, and cerebellar ataxia. Due to the lack of an efficient synthetic method, commercial production of 5-HTP is only achieved by extracting from the seeds of Griffonia Smplicifolia. This study reports efficient microbial production of 5-HTP via metabolically engineered Escherichia coli. Firstly, human tryptophan hydroxylase I (TPH1) gene was functionally expressed. For endogenous supply of the cofactor tetrahydrobiopterin (BH4), human BH4 biosynthesis and regeneration pathway was reconstituted. Whole-cell bioconversion resulted in high-level production of 5-HTP (~1.2 g/L) from 2 g/L L-tryptophan in shake flasks. Further metabolic engineering efforts were employed to achieve 5-HTP biosynthesis from simple carbon sources. The whole biosynthetic pathway was divided into three functional modules, L-tryptophan module, the hydroxylation module, and the BH4 module. By reducing the copy number of L-tryptophan module, replacing TPH1 with a more stable mutant form, and promoter regulation of the BH4 module, 5-HTP was produced at a final titer of 1.3 g/L in the shake flask and 5.1 g/L in a fed-batch fermenter with glycerol as the carbon source, both of which were the highest ever reported for microbial production of 5-HTP.

Chaperone Copolymer-Assisted Catalytic Hairpin Assembly for Highly Sensitive Detection of Adenosine.

Adenosine is an endogenous molecule that plays a vital role in biological processes. Research indicates that abnormal adenosine levels are associated with a range of diseases. The development of sensors capable of detecting adenosine is pivotal for early diagnosis of disease. For example, elevated adenosine levels are closely associated with the onset and progression of cancer. In this study, we designed a novel DNA biosensor utilizing chaperone copolymer-assisted catalytic hairpin assembly for highly sensitive detection of adenosine. The functional probe comprises streptavidin magnetic beads, an aptamer, and a catalytic chain. In the presence of adenosine, it selectively binds to the aptamer, displacing the catalytic chain into the solution. The cyclic portion of H1 hybridizes with the catalytic strand, while H2 hybridizes with the exposed H1 fragment to form an H1/H2 complex containing a G-quadruplex. Thioflavin T binds specifically to the G-quadruplex, generating a fluorescent signal. As a nucleic acid chaperone, PLL-g-Dex expedites the strand exchange reaction, enhancing the efficiency of catalytic hairpin assembly, thus amplifying the signal and reducing detection time. The optimal detection conditions were determined to be a temperature of 25 °C and a reaction time of 10 min. Demonstrating remarkable sensitivity and selectivity, the sensor achieved a lowest limit of detection of 9.82 nM. Furthermore, it exhibited resilience to interference in complex environments such as serum, presenting an effective approach for rapid and sensitive adenosine detection.

Accuracy of dual-energy computed tomography for bone marrow edema in the sacroiliac joint: A systematic review and meta-analysis.

BACKGROUND: This systematic literature review and meta-analysis aimed to assess the accuracy, sensitivity, and specificity of dual-energy computed tomography (DECT) of the sacroiliac joint. Bone marrow edema (BME) of the sacroiliac joint is an early manifestation of some diseases, such as ankylosing spondylitis, and is usually examined by nuclear magnetic resonance imaging (MRI); however, MRI can be intolerable for some patients; hence, numerous studies have analyzed DECT examinations. METHODS: We searched PUBMED, CNKI, and EMBASE in 2023 for articles containing the following terms (DECT) or (DE-CT) or (dual-energy CT) or "dual-energy CT" or (dual-energy computed tomography) and ((sacroiliac joint) or (ankylosing spondylitis) or (sacroiliac arthritis) or (sacroiliitis)). An initial search identified 444 articles, of which 7 met the criteria. Data were extracted to calculate the sensitivity, specificity, and diagnostic odds for analysis using R software. RESULTS: Out of 291 patients and 577 sacroiliac joints, 429 (74.35%) exhibited BME. All studies used magnetic resonance as the control group. The overall sensitivity and specificity of DECT were 79%, and 92%, respectively, with positive prediction rate of 92.55% and negative prediction rate of 83.73%. CONCLUSION: DECT appears to be a promising diagnostic tool for detecting BME in the sacroiliac joint and can be used as an alternative examination method for patients in whom MRI is contraindicated.

Higher serum ferritins are associated with higher blood pressure: A cross-sectional study.

The aim of the study was to investigate the association between serum ferritin and hypertension among American adults from National Health and Nutrition Examination Survey (NHANES) 1999 to 2018. A total of 16,125 participants were included. Weighted logistic regression and subgroup analyses were performed to explore the association. We found that serum ferritin was closely correlated to hypertension. Individuals with high serum ferritin were more likely to have higher systolic or diastolic blood pressure (SBP, DBP) than those with lower serum ferritin. Restricted cubic spline showed a significant non-linear association between serum ferritin and SBP/DBP. Higher level of serum ferritin (Q3 74.1-147 µg/L and Q4 > 147 µg/L) was found to have positive association with high SBP [Q3 (OR: 1.246, 95% CI:1.020-1.523), Q4 (OR: 1.354, 95% CI:1.096-1.674)], and hypertension [Q3 (OR: 1.283, 95% CI:1.099-1.499), Q4 (OR: 1.424, 95% CI:1.197-1.63)] in the whole population. In people aged between 20 and 60, subjects with high serum ferritin were significantly associated with a higher risk of hypertension, but in those over 60, the relationship between serum ferritin level and hypertension is negative. A non-linear association between serum ferritin and SBP, as well as DBP, was discovered. There was age difference in association between serum ferritin and hypertension in American adults, and further researches were needed to understand the mechanisms behind the difference.

A facile synthesis of 5-amino-[1,2,3]triazolo[5,1-a]isoquinoline derivatives through copper-catalyzed cascade reactions.

A copper-catalyzed cascade method was developed for the synthesis of [1,2,3]triazolo[5,1-a]isoquinoline derivatives. The N-fused heterocycles were obtained in good to excellent yields under mild conditions, which provided a new strategy for the preparation of this highly functionalized building block.

Logistics hub location for high-speed rail freight transport with road-rail intermodal transport network.

In this paper, we propose a new freight mode to describe how the designed HSR freight train serve for express delivery. We introduce the functions of the hubs and design the hybrid hub-and-spoke network of road-rail intermodal transportation from the perspective of planners, which features as single allocation rule and configures varying levels of hubs. The problem is accurately described by a mixed integer programming model with the objective to minimize total construction cost and total operation cost. We develop a hybrid heuristic algorithm based on a greedy strategy to obtain the levels of hubs, customer allocation and cargo routing. Taking the example of HSR freight network consisting of 50 cities in China, numerical experiments are conducted on the basis of forecasting data from the real-life express market to obtain the hub location schemes. The validity of the model and the performance of the algorithm are verified.

Incidence and risk factors of in-hospital prosthesis-related complications following total shoulder arthroplasty.

BACKGROUND: The occurrence of prosthesis-related complications after total shoulder arthroplasty is devastating and costly. The purpose was to determine the incidence and risk of in-hospital prosthesis-related complications after total shoulder arthroplasty utilizing a large-scale sample database. METHODS: A retrospective database analysis was performed based on Nationwide Inpatient Sample from 2010 to 2014. Patients who underwent total shoulder arthroplasty were included. Patient demographics, hospital characteristics, length of stay, economic indicators, in-hospital mortality, comorbidities, and peri-operative complications were evaluated. RESULTS: A total of 34,198 cases were capture from the Nationwide Inpatient Sample database. There were 343 cases of in-hospital prosthesis-related complications after total shoulder arthroplasty and the overall incidence was 1%, with a more than 2.5-fold decrease from 2010 to 2014. Dislocation was the most common category among prosthesis-related complications (0.1%). The occurrence of in-hospital prosthesis-related complications was associated with significantly more total charges and slightly longer length of stay while less usage of Medicare. Risk factors of prosthesis-related complications were identified including younger age (<64 years), female, the native American, hospital in the South, alcohol abuse, depression, uncomplicated diabetes, diabetes with chronic complications, fluid and electrolyte disorders, metastatic cancer, neurological disorders, and renal failure. Interestingly, advanced age (≥65 years) and proprietary hospital were found as protective factors. Furthermore, prosthesis-related complications were associated with aseptic necrosis, rheumatoid arthritis, rotator cuff tear arthropathy, Parkinson's disease, prior shoulder arthroscopy, and blood transfusion. CONCLUSIONS: It is of benefit to study risk factors of prosthesis-related complications following total shoulder arthroplasty to ensure the appropriate management and optimize consequences although a relatively low incidence was identified.

Naturalization of an alien ancient fruit tree at a fine scale: Community structure and population dynamics of Cydonia oblonga in China.

Naturalized plants play pivotal roles in local plant biodiversity and ecological functions; however, the drivers of naturalization remain poorly understood at a fine scale. Thus, understanding the processes of the development and dominance of alien plants in local natural habitats is of paramount importance. In the present study, we report for the first time the naturalization of Cydonia oblonga in China based on community structure and population dynamics at a fine scale. We conducted a comprehensive survey of the species through field community investigations, interviews, and a literature review. Cydonia oblonga is an ancient fruit tree with a long introduction history of over 4500 years worldwide and a cultivation history of over 2500 years in China. We analyzed C. oblonga community structure using the spatiotemporal substitution method and quantitatively analyzed population dynamics using a static life table, survivorship curve, and time series model to explore the naturalization processes. The following results were obtained. (i) The community comprised 31 coexisting vascular plant species (16 woody and 15 herbaceous species) belonging to 28 genera in 20 families. Rosaceae and Asteraceae were the two most dominant families. (ii) All individuals in the shrub layer as well as the C. oblonga population exhibited a roughly inverted J-shaped basal diameter distribution. A complete age structure was noted, and the survival curve was classified as Deevey type II. According to time series analysis, the population is estimated to increase in the future, specifically of medium and large individuals. (iii) Religious exchange, potent resource competitiveness, and similarity with the native habitat may be the major drivers of the introduction and successful naturalization of C. oblonga. These results suggest that alien species closely related to native ones are more likely to invade, naturalize, and dominate communities in local habitats.

The Amorphous Solid Dispersion of Chrysin in Plasdone® S630 Demonstrates Improved Oral Bioavailability and Antihyperlipidemic Performance in Rats.

Chrysin is a flavonoid with various biological activities. However, its low water solubility and strong metabolism render its oral bioavailability rather poor. This study aimed to develop a stable solid dispersion formulation of chrysin to improve the dissolution of chrysin, so as to increase its oral bioavailability and improve its antihyperlipidemic activities. A solid dispersion of chrysin was prepared using a solvent evaporation method, with Plasdone® S630 as the hydrophilic carrier. The formulations were characterized via X-ray diffraction, in vitro dissolution studies, and stability studies. An in-situ perfusion model was used to evaluate the absorption rates. Plasma pharmacokinetics and antihyperlipidemic performance after the oral administration of the chrysin formulations were investigated in rats. It was found that the solid dispersion of chrysin prepared using the drug-polymer mass ratio of 1:6 can form the optimized formulation. X-ray diffraction results showed that the chrysin was in an amorphous state in this optimized formulation. The cumulative release percentage of the optimized solid dispersion of chrysin at pH 1.2 and pH 6.8 was elevated to above 90% within 24 h, indicating that the formulation could enhance the dissolution rates of chrysin. Stability studies showed that the optimized formulation presented acceptable long-term storage stability, but it was susceptible to high temperature and humidity. The solid dispersion of chrysin showed higher absorption rates in the in-situ perfusion model. Pharmacokinetic studies revealed that Cmax and AUC after the intragastric administration of solid dispersion of chrysin were appreciably higher than those resulting from chrysin suspension. The oral bioavailability of the solid dispersion of chrysin was 41 times higher than that of chrysin suspension. Pharmacological studies suggested that the solid dispersion of chrysin was more powerful than chrysin raw material in improving biochemical indicators in the hyperlipidemic model in rats. This study reveals the potential use of a novel oral formulation of chrysin to reduce the currently required high dose.

Overcoming Suppressive Tumor Microenvironment by Vaccines in Solid Tumor.

Conventional vaccines are widely used to boost human natural ability to defend against foreign invaders, such as bacteria and viruses. Recently, therapeutic cancer vaccines attracted the most attention for anti-cancer therapy. According to the main components, it can be divided into five types: cell, DNA, RNA, peptide, and virus-based vaccines. They mainly perform through two rationales: (1) it trains the host immune system to protect itself and effectively eradicate cancer cells; (2) these vaccines expose the immune system to molecules associated with cancer that enable the immune system to recognize and destroy cancer cells. In this review, we thoroughly summarized the potential strategies and technologies for developing cancer vaccines, which may provide critical achievements for overcoming the suppressive tumor microenvironment through vaccines in solid tumors.