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.

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.

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.

Discovery of a Novel Photocaged PI3K Inhibitor Capable of Real-Time Reporting of Drug Release.

A novel photocaged PI3K inhibitor 2 was designed and synthesized by introducing a cascade photocaging group to block its key interaction with the kinase. Upon UV light irradiation, the photocaged compound released a highly potent PI3K inhibitor to recover its anticancer properties and a fluorescent dye for real-time reporting of drug release, providing a new approach for studying the PI3K signaling transduction pathway as well as developing precisely controlled cancer therapeutics.

Relationships between antibiotic exposure and asthma in adults in the United States: results of the National Health and Nutrition Examination Survey between 1999 and 2018.

Objectives: To investigate the relationship between antibiotic exposure and asthma in adults in the United States. Methods: Data was obtained from the National Health and Nutrition Examination Survey (NHANES) conducted between 1999 and 2018. A total of 51,124 participants were included, excluding those who were aged < 20 years, female participants who were pregnant, and individuals who did not complete the prescription medications questionnaire and the medical conditions questionnaire regarding asthma status. Antibiotic exposure was defined as the utilization of antibiotics within the past 30 days, categorized based on the Multum Lexicon Plus therapeutic classification system. Asthma was defined as having a history of asthma or having an asthma attack or wheezing symptoms in the past year. Results: The risk of asthma was found to be 2.557 (95% CI: 1.811, 3.612), 1.547 (95% CI: 1.190, 2.011) and 2.053 (95% CI: 1.344, 3.137) times greater in participants who had used macrolide derivatives, penicillin and quinolones in the past 30 days, respectively, compared with those not using antibiotics. After adjusting for demographic covariates and asthma-related factors, only macrolides derivatives were significantly associated with asthma in the 20-40 and 40-60 age groups. For individuals over 60 years old, quinolones were significantly associated with asthma. The effect of different types of antibiotic with asthma varied in male and female populations. Moreover, higher socioeconomic status, greater BMI, younger age, smoking habits, history of infection, chronic bronchitis, emphysema, and family history of asthma were all identified as risk factors for asthma. Conclusion: Our study indicated that three types of antibiotics were significantly associated with asthma in different subgroups of the population. Therefore, the use of antibiotics should be more strictly regulated.

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.

The role of shoot-derived RNAs transported to plant root in response to abiotic stresses.

A large number of RNA molecules are transported over long-distance between shoots and roots via phloem in higher plants. Mobile RNA signals are important for plants to tackle abiotic stresses. Shoot-derived mobile RNAs can be involved in the response to different developmental or environmental signals in the root. Some environmental conditions such as climate change, water deficit, nutrient deficiency challenge modern agriculture with more expeditious abiotic stress conditions. Root architecture determines the ability of water and nutrient uptake and further abiotic stress tolerance, and shoot tissue also determines the balance between shoot-root relationship in plant growth and adaptations. Thus, it is necessary to understand the roles of shoot-derived RNA signals and their potential function in roots upon abiotic stresses in the model plants (Arabidopsis thaliana and Nicotiana benthamiana) and agricultural crops. In this review, we summarize the so-far discovered shoot-derived mobile RNA transportation to the root under abiotic stress conditions, e.g. drought, cold stress and nutrient deficiencies. Furthermore, we will focus on the biological relevance and the potential roles of these RNAs in root development and stress responses which will be an asset for the future breeding strategies.

ECSIT inhibits cell death to increase tumor progression and metastasis via p53 in human breast cancer.

Background: Currently, breast cancer has surpassed lung cancer as the most common cancer and the molecular mechanism involved in tumor initiation and metastasis was unclear. Therefore, it is necessary to advance our understanding of tumor progression and metastasis and find out new targets. An evolutionarily conserved signaling intermediate in Toll pathways (ECSIT) is involved in the innate immune response and has been shown as tumor suppressors by downregulating nuclear factor-kappa B (NF-κB) pathway. However, the role of ECSIT in the progression and metastasis of human breast cancer remains unknown. Methods: We overexpressed ECSIT by transfection of a eukaryotic expression plasmid and constructed a breast cancer cell line with stable knockdown of ECSIT by short hairpin RNA. And we silenced p53 through small interfering RNA. In vivo, we replicated a xenograft mouse model in nude mice. The effects on the proliferation, viability, migration and invasion were studied by 5-ethynyl-2-deoxyuridine, cell counting Kit-8, wound healing and invasion assays. Propidium iodide/Hoechst 33342 staining and cleaved-caspase-3 staining were used to verify cell death. Western blot, immunohistochemistry (IHC) and histological analyses were used to explore the regulatory mechanism of tumor changes. Results: We reported the association of ECSIT with human breast cancer. In vitro assays demonstrated that ECSIT promoted MDA-MB-231 cell proliferation (by 66.15%), migration and invasion (by 58.29%). Knockdown of ECSIT significantly decreased cell proliferation (by 38.33%), viability, migration and invasion (by 62.37%), and increased cell death (by 41.1%). The in vivo results further confirmed that knockdown of ECSIT depressed tumorigenicity (by 29.46%) and metastasis (by 76.19%). Mechanistic investigations indicated that silencing of ECSIT could decrease the expression of p65 (by 46.05%), a subunit of NF-κB, and increase p53 protein expression in nuclei (by 89.53%). Moreover, we demonstrated that knockdown of p53 abolished the protection against cell death, which indicated that ECSIT might be involved in breast cancer progression through a p53-dependent pathway. Conclusions: Our studies provide new insight into the mechanisms underlying the role of ECSIT as well as a novel target for human breast cancer, and the development of novel ECSIT inhibitors is important for the management of TNBC.

RNA Motifs and Modification Involve in RNA Long-Distance Transport in Plants.

A large number of RNA molecules have been found in the phloem of higher plants, and they can be transported to distant organelles through the phloem. RNA signals are important cues to be evolving in fortification strategies by long-distance transportation when suffering from various physiological challenges. So far, the mechanism of RNA selectively transportation through phloem cells is still in progress. Up to now, evidence have shown that several RNA motifs including Polypyrimidine (poly-CU) sequence, transfer RNA (tRNA)-related sequence, Single Nucleotide Mutation bound with specific RNA binding proteins to form Ribonucleotide protein (RNP) complexes could facilitate RNA mobility in plants. Furthermore, some RNA secondary structure such as tRNA-like structure (TLS), untranslation region (UTR) of mRNA, stem-loop structure of pre-miRNA also contributed to the mobility of RNAs. Latest researchs found that RNA methylation such as methylated 5' cytosine (m5C) played an important role in RNA transport and function. These studies lay a theoretical foundation to uncover the mechanism of RNA transport. We aim to provide ideas and clues to inspire future research on the function of RNA motifs in RNA long-distance transport, furthermore to explore the underlying mechanism of RNA systematic signaling.

[The role of IL4I1 in immunoregulation: An update].

Interleukin-4 induced 1 protein (IL4I1), a secreted amino acid oxidase produced by antigen presenting cells, oxidizes phenylalanine to phenylpyruvate. It has been found that IL4I1 exerts an immunosuppressive function by inhibiting the proliferation and differentiation of T cells as well as limiting the proliferation of B cells. IL4I1 is involved in host defense against infection. As a gene related to poor prognosis in cancers, IL4I1 participates in tumor immune escape. IL4I1 promotes remyelination via regulation of the different phenotypes of microglia in the autoimmune demyelinating diseases, but the detailed mechanism still remains unknown. We summarize the role and mechanism of IL4I1 in the immune regulation to provide new ideas for the treatment of infections, cancers and autoimmune diseases.

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.

Systemic Long-Distance Signaling and Communication Between Rootstock and Scion in Grafted Vegetables.

Grafting is widely used in fruit, vegetable, and flower propagation to improve biotic and abiotic stress resistance, yield, and quality. At present, the systemic changes caused by grafting, as well as the mechanisms and effects of long-distance signal transport between rootstock and scion have mainly been investigated in model plants (Arabidopsis thaliana and Nicotiana benthamiana). However, these aspects of grafting vary when different plant materials are grafted, so the study of model plants provides only a theoretical basis and reference for the related research of grafted vegetables. The dearth of knowledge about the transport of signaling molecules in grafted vegetables is inconsistent with the rapid development of large-scale vegetable production, highlighting the need to study the mechanisms regulating the rootstock-scion interaction and long-distance transport. The rapid development of molecular biotechnology and "omics" approaches will allow researchers to unravel the physiological and molecular mechanisms involved in the rootstock-scion interaction in vegetables. We summarize recent progress in the study of the physiological aspects (e.g., hormones and nutrients) of the response in grafted vegetables and focus in particular on long-distance molecular signaling (e.g., RNA and proteins). This review provides a theoretical basis for studies of the rootstock-scion interaction in grafted vegetables, as well as provide guidance for rootstock breeding and selection to meet specific demands for efficient vegetable production.

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.

[Treatment of unstable pelvic fractures by cannulated screw internal fixation with the assistance of three-dimensional printing insertion template].

Objective: To evaluate the effectiveness of unstable pelvic fractures treated by cannulated screw internal fixation with the assistance of three-dimensional (3D) printing insertion template. Methods: The clinical data of 10 patients who underwent surgical treatment for unstable pelvic fractures by cannulated screw internal fixation with the assistance of 3D printing insertion template between May 2015 and June 2016 were retrospectively analysed. There were 7 males and 3 females with an average age of 37.5 years (range, 20-58 years). The causes of injury included falling from height in 5 cases, crushing from heavy load in 1 case, and traffic accidents in 4 cases. The interval from injury to admission was 1-5 hours (mean, 3.1 hours). The fracture situation included 6 cases of sacral fracture, 1 case of right sacroiliac joint dislocation, and 3 cases of iliac bone fracture. There were 10 cases of superior and inferior pubic rami fracture, including 3 cases on the left side (2 cases of suprapubic fracture adjacent to symphysis pubis), 2 cases on the right side, and 5 cases on the bilateral. All fractures were classified according to the Tile system, there were 4 cases of type B2, 1 of type B3, 4 of type C1, and 1 of type C2. The radiological outcome was evaluated by Matta scale, and the positions of the iliosacral screw and superior pubic ramus screw were evaluated according to 3D reconstruction of CT postoperatively. The functional outcome was evaluated by Majeed function scale. Results: The average time of each screw implantation was 30 minutes, and the average blood loss per screw incision was 50 mL. The time of implantation of each sacroiliac screw was 24-96 seconds (mean, 62 seconds), and the time of implantation of each suprapubic screw was 42-80 seconds (mean, 63.2 seconds). The hospitalization duration was 17-90 days (mean, 43.7 days). All incisions healed by first intention. All patients were followed up 12-22 months (mean, 15.6 months). The radiological outcome was excellent in 8 cases and good in 2 cases according to Matta scale; and 3D reconstruction of CT demonstrated that all the 9 iliosacral screws were placed as type Ⅰ, and all the 13 suprapubic ramus screws were placed as grade 0 on the first postoperative day. No complication such as neurovascular injury, screw back out or rupture, or secondary fracture displacement was observed during the follow-up. At 6 months after operation, the X-ray films showed good fracture healing in all the 10 patients. The functional outcome was excellent in 9 cases and good in 1 case according to Majeed scale at 1 year after operation. One patient sustained Tile C2 pelvic disruption complicated with L 5 nerve root injury achieved complete nervous functional recovery at last follow-up. Conclusion: It has advantages of precise screw insertion and lower risk of neurovascular injury to treat unstable pelvic fractures by cannulated screw internal fixation with the assistance of 3D printing insertion template, which can be a good alternative for the treatment of unstable pelvic fractures.

Comparative analysis of ovine adenovirus 287 and human adenovirus 2 and 5 based on their codon usage.

Ovine adenovirus 287 (OAdV287) emerges as one of the most promising gene vectors resulting from its unique biological characteristics. To obtain a more detailed knowledge about the codon usage of OAdV287, a comparative study based on the codon usage of OAdV287 and the prototypes of human adenovirus serotypes 2 and 5 (HAdV2/5) was carried out. Some commonly used indices measuring the codon usage patterns, including effective number of codons, relative synonymous codon usage, and statistical methods, were adopted. Overall, OAdV287 had a more biased and conservative codon usage pattern than that of HAdV2/5. Both mutation pressure and natural selection played important roles in shaping the codon usage patterns of these three adenoviruses. All the preference codons of OAdV287 had A/U ends and were totally different from those of sheep and humans; however, the preference codons of HAdV2/5 mostly had G/C ends and were mostly coincident with those of sheep and humans. The codon usage analysis in this study supplies some clues for further comprehending the unique biological characteristics of OAdV287 as gene vectors.

Preparation and characterization of neutralizing monoclonal antibodies against FMDV serotype O with synthetic peptide antigen.

A short linear peptide was designed according to the antigenic site analysis of VP1 protein of foot-and-mouth virus (FMDV) serotype O and synthesized as the peptide immunogen. The peptide, which covers the region from amino acid 133 to 160 of VP1 of FMDV, was linked to the N-terminal cysteine and conjugated with the carrier protein of keyhole limpet hemocyanin (KLH). Normal 6- to 8-week-old BALB/c mice were immunized with the 20 µg dose conjugated peptide antigen four times. The splenocytes from the immunized mice were fused with SP2/0 mouse myeloma cells, and positive hybridomas were selected by indirect enzyme-linked immunosorbent assay (ELISA) and subcloned four times with limiting dilution. Five stable hybridoma cell lines, designated as 4F9, 1B11, 1E10, 1D4, and 4B8, were obtained. Isotyping of all obtained MAbs indicated that the MAbs of 4F9, 1E10, and 4B8 belonged to IgG2b; the 1B11 and 1D4 belonged to IgG1 and IgM, respectively. The micro-neutralization test indicated that the MAbs of 4F9, 4B8, and 1B11 were capable of neutralizing FMDV serotype O with neutralization indices ranging from 1.81 to 2.11. These results suggest that linear synthetic peptide conjugate can elicit antibodies against native FMDV virus and can be used as an alternative immunogen for production of MAbs with exact epitope.