Fetal cerebral arteries in a new ultrasonographic perspective.

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A CD36-dependent non-canonical lipid metabolism program promotes immune escape and resistance to hypomethylating agent therapy in AML.

Environmental lipids are essential for fueling tumor energetics, but whether these exogenous lipids transported into cancer cells facilitate immune escape remains unclear. Here, we find that CD36, a transporter for exogenous lipids, promotes acute myeloid leukemia (AML) immune evasion. We show that, separately from its established role in lipid oxidation, CD36 on AML cells senses oxidized low-density lipoprotein (OxLDL) to prime the TLR4-LYN-MYD88-nuclear factor κB (NF-κB) pathway, and exogenous palmitate transfer via CD36 further potentiates this innate immune pathway by supporting ZDHHC6-mediated MYD88 palmitoylation. Subsequently, NF-κB drives the expression of immunosuppressive genes that inhibit anti-tumor T cell responses. Notably, high-fat-diet or hypomethylating agent decitabine treatment boosts the immunosuppressive potential of AML cells by hijacking CD36-dependent innate immune signaling, leading to a dampened therapeutic effect. This work is of translational interest because lipid restriction by US Food and Drug Administration (FDA)-approved lipid-lowering statin drugs improves the efficacy of decitabine therapy by weakening leukemic CD36-mediated immunosuppression.

Amplification editing enables efficient and precise duplication of DNA from short sequence to megabase and chromosomal scale.

Duplication is a foundation of molecular evolution and a driver of genomic and complex diseases. Here, we develop a genome editing tool named Amplification Editing (AE) that enables programmable DNA duplication with precision at chromosomal scale. AE can duplicate human genomes ranging from 20 bp to 100 Mb, a size comparable to human chromosomes. AE exhibits activity across various cell types, encompassing diploid, haploid, and primary cells. AE exhibited up to 73.0% efficiency for 1 Mb and 3.4% for 100 Mb duplications, respectively. Whole-genome sequencing and deep sequencing of the junctions of edited sequences confirm the precision of duplication. AE can create chromosomal microduplications within disease-relevant regions in embryonic stem cells, indicating its potential for generating cellular and animal models. AE is a precise and efficient tool for chromosomal engineering and DNA duplication, broadening the landscape of precision genome editing from an individual genetic locus to the chromosomal scale.

Engineered E. coli for D-pantothenic acid production with an acetolactate isomeroreductase mutant.

D-Pantothenic acid, as a momentous vitamin, is extensively applied to feed, medicine, cosmetics and other fields. However, there are still limitations to produce D-pantothenic acid by microbial fermentation at present. In this paper, we constructed a recombinant strain for D-pantothenic acid production by blocking the organic acid pathway, boosting pyruvate biosynthesis, relieving feedback inhibition of acetolactate synthase, improving glucose intake capacity, and modifying essential genes in the metabolic pathway. In addition, a new acetolactate isomeroreductase mutant V412A origin from Escherichia coli (EcAHAIR) encoded by ilvC was obtained to explore its substrate promiscuity. Compared with the wild type, the variant EcAHAIR-V412A has reduced steric hindrance and enhanced intermolecular forces, resulting in a high affinity for 2-acetolactate. Eventually, the fermentation production of the final strain DPAN19/trc-ilvCV412A reached 4.65 g/L, increased by 192.5% compared with strain DPA8 in shake flask cultivation and produced 62.82 g/L D-pantothenic acid in a 5 L bioreactor. The metabolic engineering strategies and enzyme modification approaches described in this paper provide a particular perspective for the bio-manufacturing of D-pantothenic acid, branched-chain amino acids and its derivates.

The interaction of Synapsin 2a and Synaptogyrin-3 regulates fear extinction in mice.

The mechanisms behind a lack of efficient fear extinction in some individuals are unclear. Here, by employing a principal components analysis-based approach, we differentiated the mice into extinction-resistant and susceptible groups. We determined that elevated synapsin 2a (Syn2a) in the infralimbic cortex (IL) to basolateral amygdala (BLA) circuit disrupted presynaptic orchestration, leading to an excitatory/inhibitory imbalance in the BLA region and causing extinction resistance. Overexpression or silencing of Syn2a levels in IL neurons replicated or alleviated behavioral, electrophysiological, and biochemical phenotypes in resistant mice. We further identified that the proline-rich domain H in the C-terminus of Syn2a was indispensable for the interaction with synaptogyrin-3 (Syngr3) and demonstrated that disrupting this interaction restored extinction impairments. Molecular docking revealed that ritonavir, an FDA-approved HIV drug, could disrupt Syn2a-Syngr3 binding and rescue fear extinction behavior in Syn2a-elevated mice. In summary, the aberrant elevation of Syn2a expression and its interaction with Syngr3 at the presynaptic site were crucial in fear extinction resistance, suggesting a potential therapeutic avenue for related disorders.

Senktide blocks aberrant RTN3 interactome to retard memory decline and tau pathology in social isolated Alzheimer's disease mice.

Sporadic or late-onset Alzheimer's disease (LOAD) accounts for more than 95% of Alzheimer's disease (AD) cases without any family history. Although genome-wide association studies have identified associated risk genes and loci for LOAD, numerous studies suggest that many adverse environmental factors, such as social isolation, are associated with an increased risk of dementia. However, the underlying mechanisms of social isolation in AD progression remain elusive. In the current study, we found that 7 days of social isolation could trigger pattern separation impairments and presynaptic abnormalities of the mossy fibre-CA3 circuit in AD mice. We also revealed that social isolation disrupted histone acetylation and resulted in the downregulation of 2 dentate gyrus (DG)-enriched miRNAs, which simultaneously target reticulon 3 (RTN3), an endoplasmic reticulum protein that aggregates in presynaptic regions to disturb the formation of functional mossy fibre boutons (MFBs) by recruiting multiple mitochondrial and vesicle-related proteins. Interestingly, the aggregation of RTN3 also recruits the PP2A B subunits to suppress PP2A activity and induce tau hyperphosphorylation, which, in turn, further elevates RTN3 and forms a vicious cycle. Finally, using an artificial intelligence-assisted molecular docking approach, we determined that senktide, a selective agonist of neurokinin3 receptors (NK3R), could reduce the binding of RTN3 with its partners. Moreover, application of senktide in vivo effectively restored DG circuit disorders in socially isolated AD mice. Taken together, our findings not only demonstrate the epigenetic regulatory mechanism underlying mossy fibre synaptic disorders orchestrated by social isolation and tau pathology but also reveal a novel potential therapeutic strategy for AD.

Design of competition nanoreactor with shell-isolated colloidal plasmonic nanomaterials for quantitative sensor platform.

Shell-isolated colloid plasmonic nanomaterials-based nanoreactor is a well-established platform widely applied in catalyst or Surface Enhanced Raman Scattering (SERS) sensors. The potentials versatility of nanoreactor platform is mainly implemented by the well-defined and tailorable structure of colloid plasmonic nanomaterials. Currently, a competitive conjugative-mediated nanoreactor is introduced to determine glucose with SERS. Glucose-conjugating nanoreactor, as convertors of the sensors, are constructed by coordinated deposition colloidal gold nanoparticles with sodium nitroprusside framework (Au@SNF) and covalently bonded 4-mercaptopyridine (4-Mpy) with self-assembly strategy. The nanoreactor contained the signal-amplifier Au@SNF NPs, conjugative-mediated signal receiver 4-Mpy, and signal internal standard molecular CN-. In addition to well-defined morphology and functionality, conjugative-mediated and internal standards method are also employed to benefit the nanoreactor. The two-parameter strategy significantly improves the signal indication and correction. Using this proposed platform, the competitive-mediated nanoreactor provides a quantitative SERS detection of glucose, and extends the applicability of SERS in more complicated and reproducibility analysis. Meanwhile, the nanoreactor based sensors also exhibited better properties to detect glucose in various food samples and bio-samples which provided strongly appliance for glucose sensors.

A fuzzy model for content-centric routing in Zigbee-based wireless sensor networks (WSNs).

WSN is one of the most efficient technologies in intelligent communication and because of its advantages, this technology has been utilized in various applications. By using WSNs, different types of data can be collected and analyzed in wide environments. The high variety of applications and types of data available in this network can cause several challenges about heterogeneous data routing. This research, presents a Fuzzy Model for Content-Centric Routing (FMCCR) in WSN to solve these challenges. The performance of FMCCR is based on two basic steps: "topology control", and "data transmission through content-centric and fuzzy logic-based routing algorithm". In the first step of FMCCR, the network topology is constructed. In the second step of the proposed method, data transmission paths are determined based on network topology and content type, and data transmission is performed. The performance of FMCCR has been evaluated in a simulation environment and the results have been compared with previous algorithms. The results show that FMCCR reduce energy consumption and improve the traffic load distribution in the network in addition to increasing the network lifetime. According to the results, FMCCR can increase network lifetime at least 10.74% and at the same time, deliver at least 88.1% more packets through the network, compared to previous methods. These results, prove the efficiency of the proposed method for using in real-world scenarios.

Impact of Atypical Hyperplasia at Surgical Margins on breast cancer outcomes in patients treated with neoadjuvant chemotherapy.

Background: Women with atypical hyperplasia (AH) is associated with a higher risk of future breast cancer. However, whether AH found at margins in patients with breast-conserving surgery (BCS) after neoadjuvant chemotherapy (NAC) needs re-excision is not well-defined. The aim of the present study was to evaluate the impact of AH at the surgical margins on the local recurrence and survival outcomes in breast cancer patients treated with NAC and BCS. Methods: A retrospective analysis comparing patients who treated with NAC and BCS with AH at the margins to those without AH was performed. Results: 598 patients were included in this study. The 5-year rates of ipsilateral breast tumor recurrence (IBTR) were 4.6% and 6.2% in patients with and without AH, respectively. No significant differences were observed among the two groups in terms of IBTR, DMFS, or OS. HER2 overexpressing breast cancer patients with severe AH at margins have a significantly higher risk of IBTR compared to those without severe AH. Conclusion: Our study suggests that the presence of AH at the surgical margins of BCS in patients who received NAC does not appear to increase the risk of ipsilateral breast cancer. Therefore, there is no need for surgeons to routinely perform additional re-excision of AH found at the margins of BCS in these patients. However, selective re-excision should be considered in certain cases, particularly in patients with HER2 overexpression.

Tau pathology epigenetically remodels the neuron-glial cross-talk in Alzheimer's disease.

The neuron-glia cross-talk is critical to brain homeostasis and is particularly affected by neurodegenerative diseases. How neurons manipulate the neuron-astrocyte interaction under pathological conditions, such as hyperphosphorylated tau, a pathological hallmark in Alzheimer's disease (AD), remains elusive. In this study, we identified excessively elevated neuronal expression of adenosine receptor 1 (Adora1 or A1R) in 3×Tg mice, MAPT P301L (rTg4510) mice, patients with AD, and patient-derived neurons. The up-regulation of A1R was found to be tau pathology dependent and posttranscriptionally regulated by Mef2c via miR-133a-3p. Rebuilding the miR-133a-3p/A1R signal effectively rescued synaptic and memory impairments in AD mice. Furthermore, neuronal A1R promoted the release of lipocalin 2 (Lcn2) and resulted in astrocyte activation. Last, silencing neuronal Lcn2 in AD mice ameliorated astrocyte activation and restored synaptic plasticity and learning/memory. Our findings reveal that the tau pathology remodels neuron-glial cross-talk and promotes neurodegenerative progression. Approaches targeting A1R and modulating this signaling pathway might be a potential therapeutic strategy for AD.

Multi-objective optimization method for coil current waveform of transcranial magnetic stimulation.

Transcranial magnetic stimulation (TMS) has been proved to be effective in the treatment of many kinds of mental diseases. However, the clicking noise produced by the pulse current with large amplitude and short duration in the TMS coil may damage the hearing of patients. The heat produced by the high-frequency pulse current in the coil also reduces the efficiency of TMS equipment. A multi-objective waveform optimization method to improve heat and noise problems at the same time is presented. By analyzing the current waveforms of TMS, the relationship between the current and the vibration energy/Joule heating is established. Taking the Joule heating and the vibration energy as the optimization objectives, exceeding the same amount of neuronal membrane potential as the limiting condition, the Pareto fronts of different current models are obtained by applying the multi-objective particle swarm optimization algorithm (MOPSO). Therefore, the corresponding current waveforms are inversely deduced. A ringing suppression cTMS (RS-cTMS) proof-of-principle experimental platform is constructed. The feasibility of the proposed method is validated through experiments. The results show that the optimized current waveforms can greatly reduce the vibration and heating of the coil compared with the conventional full-sine, recified sine and half-sine waveforms, thus reducing the pulse noise and prolonging the using time of the equipment. The optimized diversified waveforms also provide a reference for the diversity of TMS.

Characteristics and influencing factors of soil preferential flow in typical stands of Karst area in southwest China.

The study of preferential flow phenomena in Karst areas and the identification of the main factors influencing preferential flow are of great importance for the recovery of local vegetation. The distribution of the dyeing solution in the vertical and horizontal directions was examined by field staining tracer test and image processing technique. We analyzed the total dyeing area ratio, matrix flow depth, preferential flow ratio, and length index as pre-ferential flow characteristic parameters, and 14 factors affecting preferential flow using grey correlation analysis. The results showed that there were two main types of preferential flow, funnel-shaped and dendritic, with lateral water movement occurring in the soil of typical Karst stands. The mean value of the dyeing area ratio of the understory in Karst areas was 19.4%, and that of the matrix flow depth, preferential flow ratio, and length index was 4.96 cm, 62.9%, and 385.5%, respectively. Among the 14 environmental factors influencing preferential flow, the initial soil moisture content had the strongest influence on the dyeing area ratio, the available potassium content had the most significant influence on the matrix flow depth, and available phosphorous content had the most significant influence on both the preferential flow ratio and the length index. The high degree of development and spatial variability of preferential flow under typical forest stands in Karst areas was strongly influenced by physical properties such as initial soil water content, while soil nutrient also exerts important influence on preferential flow.

Performance Optimization for a Class of Petri Nets.

Petri nets (PNs) are widely used to model flexible manufacturing systems (FMSs). This paper deals with the performance optimization of FMSs modeled by Petri nets that aim to maximize the system's performance under a given budget by optimizing both quantities and types of resources, such as sensors and devices. Such an optimization problem is challenging since it is nonlinear; hence, a globally optimal solution is hard to achieve. Here, we developed a genetic algorithm combined with mixed-integer linear programming (MILP) to solve the problem. In this approach, a set of candidate resource allocation strategies, i.e., the choices of the number of resources, are first generated by using MILP. Then, the choices of the type and the cycle time of the resources are evaluated by MILP; the promising ones are used to spawn the next generation of candidate strategies. The effectiveness and efficiency of the developed methodology are illustrated by simulation studies.

Research hotspots and frontiers of childhood hypertension / 中国学校卫生

Objective@#To analyze the research status, hotspots and frontier progress of hypertension in children in English and Chinese literature, so as to provide reference for the early prevention of hypertension in children.@*Methods@#The Web of Science core collection database and CNKI database were searched to collect the literature related to the study of hypertension in children from 2000 to 2021, and the CiteSpace 5.8.R3 and VOSviewer 1.6.18 visualization tools were used to analyze the literature characteristics including publications, authors, regions, institutional cooperation, research hotspots and frontiers.@*Results@#A total of 22 687 English studies and 4 440 Chinese studies were finally included. According to the analysis results, the number of articles published on hypertension in children was on the rise. The published English articles were mainly University of Toronto and University of Colorado. The main publishing institution of Chinese articles was the First Affiliated Hospital of Peking University. The United States and China took the lead in the number of core journals published in the field of hypertension in children, the United States ranked first in terms of the influence of publications. Keyword co occurrence analysis showed that the high frequency keywords in the English literature included prevalence, risk, obesity, risk factor, body mass index, insulin resistance, overweight, metabolic syndrome, cardiovascular disease and mortality. Chinese high frequency keywords in the literature include obesity, risk factors, adiposis, influencing factors, overweight, prevalence, diabetes, treatment, health education and body mass index. The analysis of keywords showed that 25 burst terms were obtained separately in English and Chinese literature.@*Conclusion@#In recent years, the research interest on hypertension in children continues to grow and keeps updated, with the research scope expanding significantly, regarding obesity, diabetes and cardiovascular diseases.

Diagnostic efficacy of prostate imaging reporting and data system version 2.1 based on multiparametric MRI in detection of clinically significant prostate cancer / 中华放射学杂志

Objective:To evaluate the diagnostic performance of the prostate imaging reporting and data system version 2.1 (PI-RADS v2.1) based on multiparametric MRI (mpMRI) in the detection of clinically significant prostate cancer (csPCa).Methods:A total of 561 patients who underwent prostate mpMRI in the First Affiliated Hospital of Guangxi Medical University from June 2015 to December 2020 due to elevated prostate specific antigen were collected ambispectively. The patients were divided into csPCa group (276 cases) and non-csPCa group (285 cases) according to pathological findings. Prostate were scored according to the PI-RADS v2.1 scoring standard by a junior and a senior radiologist. The prostate volume was measured and the prostate specific antigen density (PSAD) was calculated. The diffusion-weighted imaging and dynamic contrast-enhanced MRI images were processed to measure the quantitative parameters of the index lesion, including apparent diffusion coefficient (ADC), volume transfer constant (K trans) and rate constant (K ep) values. The Mann-Whitney U test was used to compare the difference in parameters between the two groups. The predictors of csPCa were screened by logistic regression analysis. Predictive model of multi-parameter was established. The receiver operator characteristic curves were used to evaluate the efficacy of PI-RADS v2.1 and the model in diagnosing csPCa, and the comparisons of area under the curve (AUC) were conducted by DeLong test. Results:Compared with non-csPCa group, the patients in csPCa group had higher PI-RADS score of senior physician, PSAD, K trans and K ep value, lower ADC value ( Z=-16.69, -12.49, -3.43, -4.67, 13.91, all P<0.001). The PI-RADS scores of senior physician (OR=3.064, 95%CI 2.428-3.866, P<0.001), PSAD (OR=1.554, 95%CI 1.170-2.064, P=0.002) and ADC value (OR=0.095, 95%CI 0.032-0.288, P<0.001) were the predictors of csPCa. The AUC of junior, senior physician PI-RADS and combined prediction model were 0.861 (95%CI 0.830-0.892), 0.895 (95%CI 0.868-0.922) and 0.923 (95%CI 0.898-0.944). The pairwise difference was statistically significant (the PI-RADS score between the junior and senior physicians Z=3.24, P=0.001, the difference between the PI-RADS score of junior physician and prediction model Z=5.54, P<0.001, the difference between the PI-RADS score of senior physician and prediction model Z=4.20, P<0.001). Conclusion:Based on mpMRI, the application of PI-RADS v2.1 by junior and senior radiologists has the high diagnostic efficacy for csPCa, and the multi-parameter model has the best diagnostic efficacy for csPCa.

Study on the Application of Traditional Chinese Auspicious Images in Environmental Art Design Based on VR Senses.

Virtual reality is high-tech involving multiple disciplines developed at the end of the 20th century. Virtual reality technology can be used in many fields, such as remote control, business, education, entertainment, visual simulation, military, art, architecture, and network. Essentially, virtual reality technology has three basic characteristics: immersion, interaction, and imagination. This study analyzes the advantages of VR technology in the specific application of environmental art design based on case studies and considers the impact of VR technology on human beings from multiple dimensions such as sociology and philosophy. First, it analyzes the application and advantages of VR technology in various stages of the environmental art design. Secondly, combined with cases, it explains the advantages of VR technology in four directions: exhibition, interior, landscape, and planning. Then, it finds out the problems in the application and their solutions. In addition, to solve these problems, you need to explore the answers from more fields such as philosophy and sociology. Then, it discusses the development trend of environmental art design under VR technology and how designers should keep up with the time. Finally, as a designer of the new era, we apply VR technology to our own design and creation practice. We also analyzed the many forms of expression of auspicious images. Through the actual research and induction of modern designers, we summed up its main forms and expressions and drew some conclusions that everyone recognized, which can provide a reference for future designers.

Aerobic Exercise Training Improves Renal Injury in Spontaneously Hypertensive Rats by Increasing Renalase Expression in Medulla.

We aimed to examine the effects of aerobic exercise training on renal function in spontaneously hypertensive rats (SHR) and elucidate their possible mechanisms. Adult male SHR and age-matched Wistar-Kyoto rats (WKY) were divided into four groups: WKY sedentary group, SHR sedentary group, low-intensity training group, and medium-intensity training group. Using molecular and biochemical approaches, we investigated the effects of 14-week training on renalase (RNLS) protein levels, renal function, and apoptosis and oxidative stress modulators in kidney tissues. In vitro, angiotensin II (Ang II)-induced human kidney proximal epithelial cells (HK-2) were treated with RNLS, and changes in apoptosis and oxidative stress levels were observed. Our results show that moderate training improved renal function decline in SHR. In addition, aerobic exercise therapy significantly increased levels of RNLS in the renal medulla of SHR. We observed in vitro that RNLS significantly inhibited the increase of Ang II-inducedapoptosis and oxidative stress levels in HK-2. In conclusion, aerobic exercise training effectively improved renal function in SHR by promoting RNLS expression in the renal medulla. These results explain the possible mechanism in which exercise improves renal injury in hypertensive patients and suggest RNLS as a novel therapy for kidney injury patients.

Difference and ratio of the cross-sectional area of median nerve at the carpal tunnel and the pronator quadratus muscle in diagnosing carpal tunnel syndrome: a cross-sectional study.

Background: At present, the most commonly used diagnostic method of carpal tunnel syndrome (CTS) is based on clinical manifestations and electrophysiology, but the electrophysiology is not cheap, invasive, and lacks the presentation of peripheral nerve conditions, which is exactly the advantage of ultrasound (US). The purpose of this study was to evaluate the accuracy and effectiveness of US in the diagnosis of CTS by calculating the cross-sectional area (CSA) at the carpal tunnel and proximally at the level of the pronator quadratus muscle., and to find an appropriate index that can be used to achieve the diagnosis in a more cost-effective manner. Methods: Forty-three wrists from 35 symptomatic CTS patients and 23 wrists from 18 asymptomatic volunteers were evaluated. Diagnosis in the CTS group was based on the American Academy of Neurology clinical diagnostic criteria. The ultrasonic probe was placed at the carpal tunnel and the distal 1/3 of the pronator muscle respectively, and the carpal tunnel cross-sectional area (CSAC) and the proximal cross-sectional area (CSAP) was calculated, with a further calculation of their difference (ΔCSA) and ratio (R-CSA). Results: There was a significant difference between the 2 groups regarding mean ± standard deviation (SD) of CSAC, CSAP, ΔCSA, and R-CSA (P<0.01). The cutoff value of 12.14 mm2 for CSAC had a sensitivity and specificity of 90.7% and 100%, respectively; the cutoff value of 1.235 mm2 for R-CSA had a sensitivity and specificity of 97.67% and 95.65%, respectively; and the cutoff value of 2.035 mm2 for ΔCSA had a sensitivity and specificity of 100% and 100%, respectively. Therefore, US was found to be an effective method for the diagnosis of CTS. Receiver operating characteristic curve (ROC) analysis of all patients showed area under the curve (AUC) was 0.9778 for CSAC, 0.9949 for R-CSA and 1.000 for ΔCSA. Conclusions: US can provide reference values for the diagnosis of CTS. CSAC, ΔCSA, and R-CSA can be used for CTS diagnosis and evaluation. The ROC curve analysis showed that among the 3 values, ΔCSA was the most useful in the diagnosis of patients with CTS. ΔCSA is considered a valid diagnostic value for CTS, as its threshold of 2.04 mm2 showed the highest sensitivity and specificity.

PLHPP2 inhibits the stemness of colorectal cancer by inactivating the Nrf2 signaling pathway.

Pleckstrin homology (PH) domain leucine-rich repeat protein phosphatase 2 (PHLPP2) is a critical regulator of cellular homeostasis and acts as a tumor suppressor in multiple human cancers. However, its exact biological function in colorectal cancer (CRC) and the underlying molecular mechanism remain poorly understood. The correlation between the transcription and protein abundance of PHLPP2 was analyzed using proteomic and corresponding transcriptional data. Immunohistochemistry was used to validate the protein expression and the role of PHLPP2 in patient prognosis. In addition, a series of experiments in vitro and in vivo were performed to investigate the underlying molecular mechanism. Immunohistochemical staining of a CRC tissue microarray revealed that PHLPP2 protein expression was significantly downregulated compared to that in adjacent normal tissues. Low expression of PHLPP2 was an independent prognostic risk factor for poor survival. A nomogram established by integrating PHLPP2 expression and traditional clinicopathological factors achieved more reliable prognostic assessment in CRC patients. Additionally, PHLPP2 overexpression suppressed CRC cell migration, invasion and stemness in vitro as well as tumorigenesis in vivo. Further experiments revealed that upregulation of PHLPP2 increased ROS levels by suppressing the Nrf2-ARE signaling pathway, which inhibited the stemness of CRC cells. Moreover, incubation with sulforaphane, a selective chemical agonist of Nrf2, reversed this inhibitory effect in CRC. PHLPP2 acts as a tumor suppressor gene in CRC by restraining the Nrf2-ARE signaling pathway and increasing ROS levels, affecting the stemness of CRC cells. These anticancer molecular mechanisms indicate PHLLPP2's significant clinical value in prognosis prediction and targeted therapy.

Efficient targeted insertion of large DNA fragments without DNA donors.

Targeted insertion of large DNA fragments holds great potential for treating genetic diseases. Prime editors can effectively insert short fragments (~44 bp) but not large ones. Here we developed GRAND editing to precisely insert large DNA fragments without DNA donors. In contrast to prime editors, which require reverse transcription templates hybridizing with the target sequence, GRAND editing employs a pair of prime editing guide RNAs, with reverse transcription templates nonhomologous to the target site but complementary to each other. This strategy exhibited an efficiency of up to 63.0% of a 150-bp insertion with minor by-products and 28.4% of a 250-bp insertion. It allowed insertions up to ~1 kb, although the efficiency remains low for fragments larger than 400 bp. We confirmed efficient insertion in multiple genomic loci of several cell lines and non-dividing cells, which expands the scope of genome editing to enable donor-free insertion of large DNA sequences.