Activating innate immune responses repolarizes hPSC-derived CAR macrophages to improve anti-tumor activity.

Generation of chimeric antigen receptor macrophages (CAR-Ms) from human pluripotent stem cells (hPSCs) offers new prospects for cancer immunotherapy but is currently challenged by low differentiation efficiency and limited function. Here, we develop a highly efficient monolayer-based system that can produce around 6,000 macrophages from a single hPSC within 3 weeks. Based on CAR structure screening, we generate hPSC-CAR-Ms with stable CAR expression and potent tumoricidal activity in vitro. To overcome the loss of tumoricidal activity of hPSC-CAR-Ms in vivo, we use interferon-γ and monophosphoryl lipid A to activate an innate immune response that repolarizes the hPSC-CAR-Ms to tumoricidal macrophages. Moreover, through combined activation of T cells by hPSC-CAR-Ms, we demonstrate that activating a collaborative innate-adaptive immune response can further enhance the anti-tumor effect of hPSC-CAR-Ms in vivo. Collectively, our study provides feasible methodologies that significantly improve the production and function of hPSC-CAR-Ms to support their translation into clinical applications.

Ribocentre-switch: a database of riboswitches.

Riboswitches are regulatory elements found in the untranslated regions (UTRs) of certain mRNA molecules. They typically comprise two distinct domains: an aptamer domain that can bind to specific small molecules, and an expression platform that controls gene expression. Riboswitches work by undergoing a conformational change upon binding to their specific ligand, thus activating or repressing the genes downstream. This mechanism allows gene expression regulation in response to metabolites or small molecules. To systematically summarise riboswitch structures and their related ligand binding functions, we present Ribocentre-switch, a comprehensive database of riboswitches, including the information as follows: sequences, structures, functions, ligand binding pockets and biological applications. It encompasses 56 riboswitches and 26 orphan riboswitches from over 430 references, with a total of 89 591 sequences. It serves as a good resource for comparing different riboswitches and facilitating the identification of potential riboswitch candidates. Therefore, it may facilitate the understanding of RNA structural conformational changes in response to ligand signaling. The database is publicly available at https://riboswitch.ribocentre.org.

Chaihu-Longgu-Muli decoction improves sleep disorders by restoring orexin-A function in CKD mice.

Introduction: Chaihu-Longgu-Muli decoction (CLMD) is a well-used ancient formula originally recorded in the "Treatise on Febrile Diseases" written by the founding theorist of Traditional Chinese Medicine, Doctor Zhang Zhongjing. While it has been used extensively as a therapeutic treatment for neuropsychiatric disorders, such as insomnia, anxiety and dementia, its mechanisms remain unclear. Methods: In order to analyze the therapeutic mechanism of CLMD in chronic renal failure and insomnia, An adenine diet-induced chronic kidney disease (CKD) model was established in mice, Furthermore, we analyzed the impact of CLMD on sleep behavior and cognitive function in CKD mice, as well as the production of insomnia related regulatory proteins and inflammatory factors. Results: CLMD significantly improved circadian rhythm and sleep disturbance in CKD mice. The insomnia related regulatory proteins, Orexin, Orexin R1, and Orexin R2 in the hypothalamus of CKD mice decreased significantly, while Orexin and its receptors increased remarkably after CLMD intervention. Following administration of CLMD, reduced neuron loss and improved learning as well as memory ability were observed in CKD mice. And CLMD intervention effectively improved the chronic inflflammatory state of CKD mice. Discussion: Our results showed that CLMD could improve sleep and cognitive levels in CKD mice. The mechanism may be related to the up-regulation of Orexin-A and increased phosphorylation level of CaMKK2/AMPK, which further inhibits NF-κB downstream signaling pathways, thereby improving the disordered inflammatory state in the central and peripheral system. However, More research is required to confirm the clinical significance of the study.

Structure and ion-dependent folding of k-junctions.

k-Junctions are elaborated forms of kink turns with an additional helix on the nonbulged strand, thus forming a three-way helical junction. Two were originally identified in the structures of Arabidopsis and Escherichia coli thiamine pyrophosphate (TPP) riboswitches, and another called DUF-3268 was tentatively identified from sequence information. In this work we show that the Arabidopsis and E. coli riboswitch k-junctions fold in response to the addition of magnesium or sodium ions, and that atomic mutations that should disrupt key hydrogen bonding interactions greatly impair folding. Using X-ray crystallography, we have determined the structure of the DUF-3268 RNA and thus confirmed that it is a k-junction. It also folds upon the addition of metal ions, though requiring a 40-fold lower concentration of either divalent or monovalent ions. The key difference between the DUF-3268 and riboswitch k-junctions is the lack of nucleotides inserted between G1b and A2b in the former. We show that this insertion is primarily responsible for the difference in folding properties. Finally, we show that the DUF-3268 can functionally substitute for the k-junction in the E. coli TPP riboswitch such that the chimera can bind the TPP ligand, although less avidly.

Design, synthesis, and biological evaluation of a series of indolone derivatives as novel FLT3 inhibitors for the treatment of acute myeloid leukemia.

FLT3-ITD mutant has been extensively studied as a drug discovery target for acute myeloid leukemia. Based on our previous discovered FLT3 inhibitor (2), a series of urea group based indolone derivatives were designed, synthesized, and biological evaluated as novel FLT3 inhibitors for the treatment of FLT3-ITD positive AML. Among them, compound LC-3 exhibited potent inhibitory effects against FLT3 (IC50 = 8.4 nM) and significantly inhibited the proliferation of FLT3-ITD positive AML cells MV-4-11 (IC50 = 5.3 nM). In the cellular context, LC-3 strongly inhibited FLT3-mediated signaling pathways and induced cellular apoptosis by arresting cell cycle in G1 phase. In the in vivo studies, LC-3 significantly suppressed the tumor growth on MV-4-11 xenograft models (10 mg/kg/day, TGI = 92.16%) without exhibiting obvious toxicity. These results suggested that compound LC-3 might be a potential drug candidate for FLT3-ITD positive AML.

Crystal structures of the NAD+-II riboswitch reveal two distinct ligand-binding pockets.

We present crystal structures of a new NAD+-binding riboswitch termed NAD+-II, bound to nicotinamide mononucleotide (NMN), nicotinamide adenine dinucleotide (NAD+) and nicotinamide riboside (NR). The RNA structure comprises a number of structural features including three helices, one of which forms a triple helix by interacting with an A5 strand in its minor-groove, and another formed from a long-range pseudoknot. The core of the structure (centrally located and coaxial with the triplex and the pseudoknot) includes two consecutive quadruple base interactions. Unusually the riboswitch binds two molecules of ligand, bound at distinct, non-overlapping sites in the RNA. Binding occurs primarily through the nicotinamide moiety of each ligand, held by specific hydrogen bonding and stacking interactions with the pyridyl ring. The mode of binding is the same for NMN, NR and the nicotinamide moiety of NAD+. In addition, when NAD+ is bound into one site it adopts an elongated conformation such that its diphosphate linker occupies a groove on the surface of the RNA, following which the adenine portion inserts into a pocket and makes specific hydrogen bonding interactions. Thus the NAD+-II riboswitch is distinct from the NAD+-I riboswitch in that it binds two molecules of ligand at separate sites, and that binding occurs principally through the nicotinamide moiety.

Ribocentre: a database of ribozymes.

Ribozymes are excellent systems in which to study 'sequence - structure - function' relationships in RNA molecules. Understanding these relationships may greatly help structural modeling and design of functional RNA structures and some functional structural modules could be repurposed in molecular design. At present, there is no comprehensive database summarising all the natural ribozyme families. We have therefore created Ribocentre, a database that collects together sequence, structure and mechanistic data on 21 ribozyme families. This includes available information on timelines, sequence families, secondary and tertiary structures, catalytic mechanisms, applications of the ribozymes together with key publications. The database is publicly available at https://www.ribocentre.org.

Intercropping affects the physiology and cadmium absorption of pakchoi, lettuce, and radish seedlings.

Intercropping can affect the growth and elemental absorption of vegetables. This study investigated the physiology and cadmium (Cd) content of pakchoi (Brassica chinensis L.), lettuce (Lactuca sativa L. var. ramosa Hort.), and radish (Raphanus sativus L.) seedlings under monoculture, mutual intercropping of two or three varieties. Intercropping is not conducive to the accumulation of chlorophyll and biomass content of pakchoi, lettuce, and radish. When three seedlings were intercropped together, the antioxidant enzyme activity of pakchoi, lettuce, and radish increased and the content of malondialdehyde decreased, except that the superoxide dismutase activity of radish is inferior to the value of radish and pakchoi intercropping. Intercropping increased the soluble sugar and proline content in the lettuce seedlings, while those in the radish and lettuce seedlings reduced or had no significant effect. When intercropped with pakchoi and lettuce, the Cd content in the roots and shoots of pakchoi is higher and lower, respectively. At the same time, root or shoot bio-concentration factors also performed the same trend, and TF was the smallest and less than 1; however, the TF of lettuce is greater than 1. When intercropping with pakchoi or lettuce separately or together, it promoted the accumulation of Cd in radish root; when intercropping with pakchoi, the value of TF was the smallest. From the antioxidant system, the performance of the three seedlings intercropped together is better than the two; however, the accumulation of Cd shows the opposite trend, and the participation of cabbage in the intercropping is relatively conducive to reducing the Cd content in the edible parts.

Dynamic Changes of Phenolic Composition, Antioxidant Capacity, and Gene Expression in 'Snow White' Loquat (Eriobotrya japonica Lindl.) Fruit throughout Development and Ripening.

The newly released 'Snow White' (SW), a white-fleshed loquat (Eriobotrya japonica Lindl.) cultivar, holds promise for commercial production. However, the specifics of the phenolic composition in white-fleshed loquats, along with the antioxidant substances and their regulatory mechanisms, are not yet fully understood. In this study, we examined the dynamic changes in the phenolic compounds, enzyme activities, antioxidant capacity, and gene expression patterns of SW during the key stages of fruit development and ripening. A total of 18 phenolic compounds were identified in SW, with chlorogenic acid, neochlorogenic acid, and coniferyl alcohol being the most predominant. SW demonstrated a stronger antioxidant capacity in the early stages of development, largely due to total phenolics and flavonoids. Neochlorogenic acid may be the most significant antioxidant contributor in loquat. A decline in enzyme activities corresponded with fruit softening. Different genes within a multigene family played distinct roles in the synthesis of phenolics. C4H1, 4CL2, 4CL9, HCT, CCoAOMT5, F5H, COMT1, CAD6, and POD42 were implicated in the regulation of neochlorogenic acid synthesis and accumulation. Consequently, these findings enhance our understanding of phenolic metabolism and offer fresh perspectives on the development of germplasm resources for white-fleshed loquats.

Accuracy of Oxygen Saturation Measurements in Patients with Obesity Undergoing Bariatric Surgery.

BACKGROUND: We aimed to determine the magnitude, direction, and influencing factors of the concordance between arterial oxygen saturation (SaO2) and peripheral capillary oxygen saturation (SpO2) in patients with obesity undergoing bariatric surgery, supporting the measurement of SaO2 and SpO2 in key populations. METHODS: Patients with obesity undergoing bariatric surgery from 2017 to 2020 were included. Preoperative SpO2 and SaO2 were collected. Linear correlation and multiple linear regression analyses were performed to characterize the relationships between body mass index (BMI), age, and sex with pulse oximetry and arterial blood gas (ABG) parameters. Bland-Altman analysis was applied to determine the concordance between SpO2 and SaO2 and the limits of this concordance. RESULTS: A total of 134 patients with obesity undergoing bariatric surgery were enrolled. SaO2 was negatively associated with BMI (p < 0.0001) and age (p = 0.006), and SpO2 was negatively associated with BMI (p = 0.021) but not with age. SpO2 overestimated SaO2 in 91% of patients with a bias of 2.05%. This bias increased by 203% in hypoxemic patients compared with nonhypoxemic patients (p < 0.0001). The bias was 1.3-fold higher (p = 0.023) in patients with a high obesity surgery mortality risk score (OS-MRS) than in those with low or intermediate scores. CONCLUSION: Compared with SpO2, preoperative SaO2 can more accurately reflect the real oxygen saturation in patients with obesity undergoing bariatric surgery, especially for those with BMI ≥ 40 kg/m2, age ≥ 40 years, and high OS-MRS. ABG analysis can provide a more reliable basis for accurate and timely monitoring, ensuring the perioperative safety of susceptible patients.

Salicylic acid alleviates selenium stress and promotes selenium uptake of grapevine.

To determine suitable cultivation measures to enrich selenium (Se) and alleviate the Se stress in fruit trees, the effects of different exogenous salicylic acid (SA) concentrations (0, 50, 100, 150 and 200 mg/L) on the growth and Se uptake of grapevine under Se stress were studied. Under Se stress, SA increased the biomass of grapevine to some extent and had a linear relationship with both root and shoot biomass. The chlorophyll content, net photosynthetic rate, transpiration rate, stomatal conductance, and intercellular CO2 concentration of grapevine tended to increase when the concentration of SA was < 150 mg/L and decrease when the concentration of SA was > 150 mg/L. Different concentrations of SA enhanced the activity of superoxide dismutase, while reducing that of peroxidase. It had no significant effect on the catalase activity of grapevine. SA decreased the content of osmotically active substances in grapevine to some extent. SA also increased the contents of total Se, inorganic Se and organic Se in grapevine to some extent, and had a linear or quadratic polynomial relationship with the total Se contents in both roots and shoots. When the SA concentration was 250 mg/L, the total Se contents in the roots and shoots were the highest and increased by 10.41% and 58.46%, respectively, compared with the control. Therefore, exogenous SA could promote the growth and Se uptake of grapevine under Se stress, with 250 mg/L serving as the most effective concentration.

Structure and mechanism of a methyltransferase ribozyme.

Known ribozymes in contemporary biology perform a limited range of chemical catalysis, but in vitro selection has generated species that catalyze a broader range of chemistry; yet, there have been few structural and mechanistic studies of selected ribozymes. A ribozyme has recently been selected that can catalyze a site-specific methyl transfer reaction. We have solved the crystal structure of this ribozyme at a resolution of 2.3 Å, showing how the RNA folds to generate a very specific binding site for the methyl donor substrate. The structure immediately suggests a catalytic mechanism involving a combination of proximity and orientation and nucleobase-mediated general acid catalysis. The mechanism is supported by the pH dependence of the rate of catalysis. A selected methyltransferase ribozyme can thus use a relatively sophisticated catalytic mechanism, broadening the range of known RNA-catalyzed chemistry.

The effect of a seven-step rehabilitation training program on cardiac function and quality of life after percutaneous coronary intervention for acute myocardial infarction.

OBJECTIVES: To study the effect of a seven-step rehabilitation training program on cardiac function and quality of life in acute myocardial infarction (AMI) patients after percutaneous coronary intervention (PCI). METHODS: In this study one hundred AMI patients undergoing emergency PCI at The First Hospital of Fangshan District between June 2019 and June 2020 were included. Patients were retrospectively divided into two equal groups based on the type of physiotherapy regiment. The training group included patients who underwent seven-step rehabilitation training while the control group had patients who received routine nursing. Left ventricular ejection fraction (LVEF), self-care capability, hospitalization duration, quality of life, and adverse cardiac event incidence were compared. RESULTS: The number of patients with LVEF values ≥ 50% was significantly higher in the training group after one week of training. Training group patients also showed decreased hospitalization duration and larger improvement in self-care capacity scores. At three months after training, training group patients had overall superior quality of life and lower incidence rates of arrhythmia and angina pectoris. CONCLUSION: The seven-step rehabilitation training program has a significant effect on improving AMI patient quality of life and cardiac function post-PCI, and is worthy of continued study and promotion.

CCDC65 as a new potential tumor suppressor induced by metformin inhibits activation of AKT1 via ubiquitination of ENO1 in gastric cancer.

The coiled-coil domain containing protein members have been well documented for their roles in many diseases including cancers. However, the function of the coiled-coil domain containing 65 (CCDC65) remains unknown in tumorigenesis including gastric cancer. Methods: CCDC65 expression and its correlation with clinical features and prognosis of gastric cancer were analyzed in tissue. The biological role and molecular basis of CCDC65 were performed via in vitro and in vivo assays and a various of experimental methods including co-immunoprecipitation (Co-IP), GST-pull down and ubiquitination analysis et al. Finally, whether metformin affects the pathogenesis of gastric cancer by regulating CCDC65 and its-mediated signaling was investigated. Results: Here, we found that downregulated CCDC65 level was showed as an unfavourable factor in gastric cancer patients. Subsequently, CCDC65 or its domain (a.a. 130-484) was identified as a significant suppressor in GC growth and metastasis in vitro and in vivo. Molecular basis showed that CCDC65 bound to ENO1, an oncogenic factor has been widely reported to promote the tumor pathogenesis, by its domain (a.a. 130-484) and further promoted ubiquitylation and degradation of ENO1 by recruiting E3 ubiquitin ligase FBXW7. The downregulated ENO1 decreased the binding with AKT1 and further inactivated AKT1, which led to the loss of cell proliferation and EMT signal. Finally, we observed that metformin, a new anti-cancer drug, can significantly induce CCDC65 to suppress ENO1-AKT1 complex-mediated cell proliferation and EMT signals and finally suppresses the malignant phenotypes of gastric cancer cells. Conclusion: These results firstly highlight a critical role of CCDC65 in suppressing ENO1-AKT1 pathway to reduce the progression of gastric cancer and reveals a new molecular mechanism for metformin in suppressing gastric cancer. Our present study provides a new insight into the mechanism and therapy for gastric cancer.

The effects of water control on the survival and growth of Alternanthera philoxeroides in the vegetative reproduction and seedling stages.

Alternanthera philoxeroides (Martius) is an infamous invasive alien plant that is widely distributed in aquatic and terrestrial habitats. To investigate the vegetative reproduction, growth, survival strategy, and the function of leaves in fragment of A. philoxeroides under different water conditions, two water control experiments were conducted with different leaf treatments: (1) water control with stolon fragments, and (2) water control with plants. The water control was subjected to five levels: I 30% soil water content, II 70% soil water content, III 97% soil water content, IV water depth of 5 cm, and V water depth of 10 cm in combination with the two leaf treatments, fragments with two leaves and fragments without leaves. Based on the results, A. philoxeroides produced a significantly higher stem length, node number, leaf number, stem biomass, leaf biomass, and total biomass in the 97% soil water content and in treatments with leaves. Additionally, the stem mass ratio increased and the root mass ratio decreased with the increase of the water content. In Exp. 1, the survival rate was the highest in the 97% water content and was 0 in the 30% water content. Therefore, the leaves of stolon fragments contribute to the vegetative reproduction and growth of A. philoxeroides. In response to different water conditions, A. philoxeroides adopts different strategies according to the resource reserves by itself, which are conducive to its survival and widespread occurrence.

Reduced antioxidant and anti-inflammatory effects of propofol at high-dose on morbidly obese patients.

This study was aimed to investigate differences in antioxidant and anti-inflammatory effects of propofol at two commonly used dosing schedules on morbidly obese patients. Twenty-two morbidly obese patients were randomly divided into two groups, namely, TBW (dosing based on total body weight) and LBW (dosing based on lean body weight) groups. Three biomarkers, i.e. superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO) were measured as indicators of the level of oxidation stress reaction. Pro-inflammatory cytokines including Interleukin-6 (IL-6) and Interleukin-8 (IL-8) were used to describe the degree of inflammation. Plasma levels of SOD, MDA and NO were increased and reached a peak value 0.5h after anesthesia induction, but the increase was smaller in the LBW group compared with the TBW group. Besides, plasma concentrations of IL-6 and IL-8 were also increased and attained a peak level 0.5h after anesthesia induction, but the increase was higher in the TBW group compared with the LBW group. The LBW-based dosing of propofol had more potent antioxidant and anti-inflammatory effects than the TBW-based dosing during anesthesia induction period on morbidly obese patients. This study provided a dosing recommendation of propofol for morbidly obese patients.

Inhibitory Top-Down Control Deficits in Schizophrenia With Auditory Verbal Hallucinations: A Go/NoGo Task.

Objective: Auditory verbal hallucinations (AVH), with unclear mechanisms, cause extreme distresses to schizophrenia patients. Deficits of inhibitory top-down control may be linked to AVH. Therefore, in this study, we focused on inhibitory top-down control in schizophrenia patients with AVH. Method: The present study recruited 40 schizophrenia patients, including 20 AVH patients and 20 non-AVH patients, and 23 healthy controls. We employed event-related potentials to investigate the N2 and P3 amplitude and latency differences among these participants during a Go/NoGo task. Results: Relative to healthy controls, the two patient groups observed longer reaction time (RT) and reduced accuracy. The two patient groups had smaller NoGo P3 amplitude than the healthy controls, and the AVH patients showed smaller NoGo P3 amplitude than the non-AVH patients. In all the groups, the parietal area showed smaller NoGo P3 than frontal and central areas. However, no significant difference was found in N2 and Go P3 amplitude between the three groups. Conclusions: AVH patients might have worse inhibitory top-down control, which might be involved in the occurrence of AVH. Hopefully, our results could enhance understanding of the pathology of AVH.

Pleural Fluid GSDMD Is a Novel Biomarker for the Early Differential Diagnosis of Pleural Effusion.

OBJECTIVE: Gasdermin D (GSDMD), controlling pyroptosis in cells, has multiple physiological functions. The diagnostic role of GSDMD in pleural effusion (PE) remains unknown. METHODS: Sandwich ELISA kits that we developed were applied to measure the level of GSDMD for 335 patients with a definite cause of PE, including transudative PE, tuberculous pleural effusion (TPE), parapneumonic pleural effusion (PPE), and malignant pleural effusion (MPE). The diagnostic accuracy of Light's criteria vs. the new marker GSDMD was performed. Clinical follow-up of 40 cases of PPE was conducted and divided into efficacy and non-efficacy groups according to the therapeutic outcome. Nucleated cells (NCs) in PE were isolated and further infected with bacteria to verify the cell source of GSDMD. RESULTS: The diagnostic accuracy of GSDMD for the diagnosis of PE were 96% (sensitivity) and 94% (specificity). The receiver operating characteristic (ROC) curve indicated that GSDMD can be an efficient biomarker for the differential diagnosis of transudative PE and other groups (all AUC > 0.973). Noteworthily, the highest AUC belonged to tuberculosis diagnosis of 0.990, and the cut-off value was 18.40 ng/mL. Moreover, the same cut-off value of PPE and MPE was 9.35 ng/mL. The combination of GSDMD, adenosine deaminase (ADA), and lactate dehydrogenase (LDH) will further improve the diagnostic efficiency especially between TPE and PPE (AUC = 0.968). The AUC of GSDMD change at day 4, which could predict the therapeutic effect at an early stage, was 0.945 (P < 0.0001). Interestingly, bacterial infection experiments further confirm that the pleural fluid GSDMD was expressed and secreted mainly by the NCs. CONCLUSION: GSDMD and its combination are candidates as a potentially novel biomarker not only to separate PEs early and effectively, but also monitor disease progression.

Structure and folding of four putative kink turns identified in structured RNA species in a test of structural prediction rules.

k-Turns are widespread key architectural elements that occur in many classes of RNA molecules. We have shown previously that their folding properties (whether or not they fold into their tightly kinked structure on addition of metal ions) and conformation depend on their local sequence, and we have elucidated a series of rules for prediction of these properties from sequence. In this work, we have expanded the rules for prediction of folding properties, and then applied the full set to predict the folding and conformation of four probable k-turns we have identified amongst 224 structured RNA species found in bacterial intergenenic regions by the Breaker lab (1). We have analyzed the ion-dependence of folding of the four k-turns using fluorescence resonance energy transfer, and determined the conformation of two of them using X-ray crystallography. We find that the experimental data fully conform to both the predicted folding and conformational properties. We conclude that our folding rules are robust, and can be applied to new k-turns of unknown characteristics with confidence.