Development of Cas13a-based therapy for cancer treatment.

Gene therapy has become a major focus of current biomedical research. CRISPR (Clustered Regularly Inter spaced Short Palindromic Repeats) systems have been extensively researched for disease treatment applications through genome editing specificity. Compared with Cas9 (CRISPR-associated proteins, Cas), a commonly used tool enzyme for genome editing, Cas13a exhibits RNA-dependent endonuclease activity, including collateral cleavage without obvious potential genetic risks. With its high specificity, Cas13a has significantly improved the sensitivity of viral diagnosis and shown potential to eliminate viruses. However, its efficacy in tumor therapy has not been determined. This review introduces the mechanism and research developments associated with the CRISPR-Cas13a system in tumor treatments and its potential to be used as a new tool for gene therapy. We hope more research would apply Cas13a-based therapy in cancer treatment in the future.

Grape SnRK2.7 Positively Regulates Drought Tolerance in Transgenic Arabidopsis.

In this study, we obtained and cloned VvSnRK2.7 by screening transcriptomic data to investigate the function of the grape sucrose non-fermenting kinase 2 (SnRK2) gene under stress conditions. A yeast two-hybrid (Y2H) assay was used to further screen for interaction proteins of VvSnRK2.7. Ultimately, VvSnRK2.7 was heterologously expressed in Arabidopsis thaliana, and the relative conductivity, MDA content, antioxidant enzyme activity, and sugar content of the transgenic plants were determined under drought treatment. In addition, the expression levels of VvSnRK2.7 in Arabidopsis were analyzed. The results showed that the VvSnRK2.7-EGFP fusion protein was mainly located in the cell membrane and nucleus of tobacco leaves. In addition, the VvSnRK2.7 protein had an interactive relationship with the VvbZIP protein during the Y2H assay. The expression levels of VvSnRK2.7 and the antioxidant enzyme activities and sugar contents of the transgenic lines were higher than those of the wild type under drought treatment. Moreover, the relative conductivity and MDA content were lower than those of the wild type. The results indicate that VvSnRK2.7 may activate the enzyme activity of the antioxidant enzyme system, maintain normal cellular physiological metabolism, stabilize the berry sugar metabolism pathway under drought stress, and promote sugar accumulation to improve plant resistance.

Transcriptome and metabolites analysis of water-stressed grape berries at different growth stages.

Drought is one of the main abiotic factors affecting grape quality. However, the impacts of drought stress on sugar and related gene expression during grape berry ripening remain unclear. In this experiment, the grapes were subjected to different levels of continuous water stress from 45 to 120 days after flowering (DAA) to study the changes in berry sugar content and the expression of genes related to sugar metabolism under different water stresses. Data supported that glucose, fructose, sucrose, and soluble sugars increased from 45 DAA. Combined with previous research results, T1, T2, and Ct grape berries with 60 ~ 75 DAA and large differences in sucrose, fructose, glucose and soluble sugars compared with the Ct were selected for RNA sequencing (RNA-seq). Through transcriptome analysis, 4471 differentially expressed genes (DEGs) were screened, and 65 genes in photosynthesis, ABA signaling pathway and photosynthetic carbon metabolism pathway were analyzed further by qRT-PCR. At 60 DAA, the relative expression levels of CAB1R, PsbP, SNRK2, and PYL9 were significantly upregulated in response to water stress, while AHK1, At4g02290 were down-regulated. At 75 DAA, the relative expression levels of ELIP1, GoLS2, At4g02290, Chi5, SAPK, MAPKKK17, NHL6, KINB2, and AHK1 were upregulated. And CAB1R, PsbA, GoLS1, SnRK2, PYL9, and KINGL were significantly downregulated under moderate water stress. In addition, PsbA expression was down-regulated in response to water stress. These results will help us to fully understand the potential connections between glucose metabolism and gene expression in grapes under drought stress.

VaSUS2 confers cold tolerance in transgenic tomato and Arabidopsis by regulation of sucrose metabolism and ROS homeostasis.

KEY MESSAGE: VaSUS2 enhances cold tolerance of transgenic tomato and Arabidopsis by regulating sucrose metabolism and improving antioxidant enzymes activity. Sucrose synthetase (SUS) is a key enzyme of sugar metabolism, and plays an important role in response to abiotic stress in plant. However, the function of VaSUS2 remains unknown in cold tolerance. Here, the cloning and functional characterization of the plasma membrane-localized VaSUS2 gene isolated from Vitis amurensis was studied. The transcript level of VaSUS2 was up-regulated under cold stress in Vitis amurensis. Heterologous expression of VaSUS2 in tomato increased SUS activity, which promoted the accumulation of glucose and fructose under cold treatment. The transgenic tomato and Arabidopsis exhibited higher levels of antioxidant enzymes activity, lower relative electrolyte leakage (REL), malondialdehyde (MDA) and hydrogen peroxide (H2O2) content compared to wild type under cold stress. Importantly, the ability of scavenging reactive oxygen species (ROS) in transgenic plants was significantly improved. Moreover, yeast two-hybrid (Y2H) indicated that VaSnRK1 might be a potential interaction protein of VaSUS2. qRT-PCR showed that sucrose metabolism-related genes SlSUS, SlSPS and SlINV were significantly up-regulated in transgenic tomatoes. Meanwhile, the expression levels of antioxidant enzyme genes and cold-related genes CBF1, COR47 and ICE1 were up-regulated in transgenic plants. Taken together, these results suggested that VaSUS2 was involved in cold tolerance by increasing the levels of soluble sugars, improving the activity of antioxidant enzymes, and up-regulating the expression of cold-related genes in transgenic tomatoes and Arabidopsis.

Overexpression VaPYL9 improves cold tolerance in tomato by regulating key genes in hormone signaling and antioxidant enzyme.

BACKGROUND: Abscisic acid (ABA) has been reported in controlling plant growth and development, and particularly dominates a role in resistance to abiotic stress. The Pyrabactin Resistance1/PYR1-Like /Regulatory Components of ABA receptors (PYR1/PYL/RCAR) gene family, of which the PYL9 is a positive regulator related to stress response in ABA signaling transduction. Although the family has been identified in grape, detailed VaPYL9 function in cold stress remains unknown. RESULTS: In order to explore the cold tolerance mechanism in grape, VaPYL9 was cloned from Vitis amurensis. The subcellular localization showed that VaPYL9 was mainly expressed in the plasma membrane. Yeast two-hybrid (Y2H) showed VaPCMT might be a potential interaction protein of VaPYL9. Through the overexpression of VaPYL9 in tomatoes, results indicated transgenic plants had higher antioxidant enzyme activities and proline content, lower malondialdehyde (MDA) and H2O2 content, and improving the ability to scavenge reactive oxygen species than wild-type (WT). Additionally, ABA content and the ratio of ABA/IAA kept a higher level than WT. Quantitative real-time PCR (qRT-PCR) showed that VaPYL9, SlNCED3, SlABI5, and antioxidant enzyme genes (POD, SOD, CAT) were up-regulated in transgenic tomatoes. Transcriptome sequencing (RNA-seq) found that VaPYL9 overexpression caused the upregulation of key genes PYR/PYL, PYL4, MAPK17/18, and WRKY in transgenic tomatoes under cold stress. CONCLUSION: Overexpression VaPYL9 enhances cold resistance of transgenic tomatoes mediated by improving antioxidant enzymes activity, reducing membrane damages, and regulating key genes in plant hormones signaling and antioxidant enzymes.

Temperature-phase transcriptomics reveals that hormones and sugars in the phloem of grape participate in tolerance during cold acclimation.

KEY MESSAGE: Most of the upregulated genes contributed to the accumulation of soluble sugars and ABA in the phloem of 'Vitis amurensis' compared to 'Merlot' during cold acclimation. Extreme cold is one of the dominant abiotic factors affecting grape yield and quality. However, the changes in sugars, phytohormones, and gene expression in the branch phloem of different tolerant grape varieties during cold acclimation remain elusive. The data supported that with decreasing temperature, the contents of fructose, sucrose, and ABA in the phloem of Vitis amurensis (cold-tolerant, T) and 'Merlot' (cold-sensitive, S) increased during cold acclimation, and these indicators were higher in T than in S. Furthermore, the activities of sucrose synthetase, sucrose phosphate synthetase, and acid invertase peaked in the early phase of cold acclimation (approximately 5 °C) compared to other phases (approximately 28 °C, 0 °C, - 5 °C and - 10 °C). Moreover, the RNA sequencing results helped identify a total of 11,343 differentially expressed genes in the phloem of T and S, among which 4912 were upregulated and 6431 were downregulated. In the abscisic acid pathway, CRTISO, PSPY1-1, CYCP707A4-2, PYL4-1, PYL4-2, P2C08, SAPK2, TARAB1, and DBF3 were more highly expressed in T than in S. In the starch and sucrose metabolism pathway, HXK1, PGMP, GLGL1, SUS6, VCINV, BGL11, SSY1, GPS, BAM1 and BAM3 were also more highly expressed in T than in S. Moreover, the genes related to oxidative phosphorylation, such as NDHF, ND4, ND1, NAD7, NAD2, ATPB, YMF19, ATP9, PMA1 and AHA8, were upregulated in T. These results will be beneficial for understanding the potential differences in tolerance across two different cold-tolerant grapes with respect to sugar metabolism and gene expression.

Existence of Classical Solutions for Nonlinear Elliptic Equations with Gradient Terms.

This paper deals with the existence of solutions of the elliptic equation with nonlinear gradient term -Δu=f(x,u,∇u) on Ω restricted by the boundary condition u|∂Ω=0, where Ω is a bounded domain in RN with sufficiently smooth boundary ∂Ω, N≥2, and f:Ω¯×R×RN→R is continuous. The existence results of classical solutions and positive solutions are obtained under some inequality conditions on the nonlinearity f(x,ξ,η) when |(ξ,η)| is small or large enough.

Genome-wide identification and expression analysis of PIN gene family under phytohormone and abiotic stresses in Vitis Vinifera L.

The auxin efflux transport proteins PIN-formed (PIN) has wide adaptability to hormone and abiotic stress, but the response mechanism of PINs in grape remains unclear. In this study, 12 members of VvPINs were identified and distributed on 8 chromosomes. The PIN genes of five species were divided into two subgroups, and the similarity of exons/introns and motifs of VvPIN genes were found in the same subgroup. Meanwhile, according to the examination of conserved motifs, the motif 3 included the conserved structure NPNTY. The promoter region of VvPIN gene family contained various cis-acting elements, which were related to light, abiotic stress, and hormones which are essential for growth and development. Additionally, VvPIN1, VvPIN9, and VvPIN11 proteins simultaneously interacted with the ARF, ABC, PINOID, GBF1, and VIT_08s0007g09010. The results of qRT-PCR revealed that the majority of the VvPINs were highly induced by NAA, GA3, ABA, MeJA, SA, NaCl, low-temperature (4 ℃), and PEG treatments, and the results were consistent with the prediction of the cis-acting elements. Moreover, the expression profile and quantitative real-time PCR (qRT-PCR) demonstrated that VvPIN genes were expressed in roots, stems, and leaves. The subcellular localization of VvPIN1 in Nicotiana benthamiana revealed that VvPIN1 was localized at the plasma membrane. Collectively, this study revealed that PIN genes could respond to various abiotic stresses, and provided a framework for regulating the expression of PIN genes to enhance the resistance of the grape. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01239-8.

A simultaneous bilateral quadriceps and patellar tendons rupture in patients with chronic kidney disease undergoing long-term hemodialysis: a case report.

BACKGROUND: The incidence of rupture of the quadriceps or patellar tendon s is low, especially that of bilateral quadriceps tendon rupture, and it is generally considered a complication secondary to chronic systemic disorders. We report two rare cases of simultaneous bilateral tendon rupture affecting the extensor function of the knee in patients with chronic kidney disease who have been treated with long-term haemodialysis. CASE PRESENTATION: Two young males with a history of chronic kidney disease who were being treated with long-term haemodialysis presented to our hospital with clinical signs of disruption of the extensor mechanism of the knee. One patient was diagnosed with bilateral quadriceps tendon rupture, and the other patient had bilateral patellar tendon rupture. They underwent surgical repair of the tendons, and their knees were actively mobilized during physiotherapy. CONCLUSION: Bilateral quadriceps or patellar tendons rupture is a rare occurrence in patients with chronic kidney disease who are being treated with long-term haemodialysis. Timely surgical treatment and scientific physiotherapy can lead to good recovery of knee joint function.

Novel [1,2,4]triazolo[1,5-a]pyrimidine derivatives as potent antitubulin agents: Design, multicomponent synthesis and antiproliferative activities.

As restricted CA-4 analogues, a novel series of [1,2,4]triazolo[1,5-a]pyrimidines possessing 3,4,5-trimethoxylphenyl groups has been achieved successfully via an efficient one-pot three-component reaction of 3-(3,4,5-trimethoxyphenyl)-1H-1,2,4-triazol-5-amine, 1,3-dicarbonyl compounds and aldehydes. Initial biological evaluation demonstrated some of target compounds displayed potent antitumor activity in vitro against three cancer cell lines. Among them, the most highly active analogue 26 inhibited the growth of HeLa, and A549 cell lines with IC50 values at 0.75, and 1.02 µM, respectively, indicating excellent selectivity over non-tumoural cell line HEK-293 (IC50 = 29.94 µM) which suggested that the target compounds might possess a high safety index. Moreover, cell cycle analysis illustrated that the analogue 26 significantly induced HeLa cells arrest in G2/M phase, meanwhile the compound could dramatically affect cell morphology and microtubule networks. In addition, compound 28 exhibited potent anti-tubulin activity with IC50 values of 9.90 µM, and molecular docking studies revealed the analogue occupied the colchicine-binding site of tubulin. These observations suggest that [1,2,4]triazolo[1,5-a]pyrimidines represent a new class of tubulin polymerization inhibitors and well worth further investigation aiming to generate potential anticancer agents.

TIMELESS contributes to the progression of breast cancer through activation of MYC.

BACKGROUND: Breast cancer is the most common malignancy and the leading cause of cancer death among women. TIMELESS (TIM), a circadian rhythm regulator, has been recently implicated in the progression of human cancer. However, the role of TIM in the progression of breast cancer has not been well-characterized. METHODS: Immunohistochemistry (IHC) staining was used to examine TIM levels in breast cancer specimens. Mammosphere formation analysis and side population analysis were used to examine the effect of TIM on the self-renewal of breast cancer stem cells. A wound healing assay and a Transwell assay were used to determine the role of TIM in breast cancer cell migration and invasion. A soft agar growth assay in vitro and tumorigenicity in vivo were used to determine the role of TIM in tumorigenicity. RESULTS: TIM levels in both breast cancer cell lines and tissues were significantly upregulated. Patients with high TIM had poorer prognosis than patients with low TIM. Overexpression of TIM dramatically enhanced, while knockdown of TIM suppressed the self-renewal of cancer stem cells (CSCs), cell invasion and migration abilities of breast cancer cells in vitro. Moreover, overexpression of TIM significantly augmented, while knockdown of TIM reduced the tumorigenicity of breast cancer cells in vivo. Mechanism studies revealed that TIM upregulated the expression and the trans-activity of the well-known oncogene MYC. Inhibition of MYC significantly blocked the effects of TIM on CSC population, cell invasion and anchor-independent cell growth. CONCLUSION: TIM plays an important role in promoting breast cancer progression and may represent a novel therapeutic target for breast cancer.

Prostate tumour overexpressed-1 promotes tumourigenicity in human breast cancer via activation of Wnt/ß-catenin signalling.

Breast cancer is the most common malignancy in females. The presence of cancer stem cells (CSCs) is the main cause of local and distant tumour recurrence and is associated with poor outcome in breast cancer. However, the molecular mechanisms underlying the maintenance of CSCs remain largely unknown. This study demonstrates that prostate tumour overexpressed-1 (PTOV1) enhances the CSC population and augments the tumourigenicity of breast cancer cells both in vitro and in vivo. Moreover, PTOV1 suppresses transcription of Dickkopf-1 (DKK1) by recruiting histone deacetylases and subsequently reducing DKK1 promoter histone acetylation, followed by activation of Wnt/ß-catenin signalling. Restoration of DKK1 expression in PTOV1-overexpressing cells counteracts the effects of PTOV1 on Wnt/ß-catenin activation and the CSC population. Collectively, these results suggest that PTOV1 positively regulates the Wnt/ß-catenin signalling pathway and enhances tumourigenicity in breast cancer; this novel mechanism may represent a therapeutic target for breast cancer. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Femtosecond Laser-Induced Phase Transformation on Single-Crystal 6H-SiC.

Silicon carbide (SiC) is widely used in many research fields because of its excellent properties. The femtosecond laser has been proven to be an effective method for achieving high-quality and high-efficiency SiC micromachining. In this article, the ablation mechanism irradiated on different surfaces of 6H-SiC by a single pulse under different energies was investigated. The changes in material elements and the geometric spatial distribution of the ablation pit were analyzed using micro-Raman spectroscopy, Energy Dispersive Spectrum (EDS), and an optical microscope, respectively. Moreover, the thresholds for structural transformation and modification zones of 6H-SiC on different surfaces were calculated based on the diameter of the ablation pits created by a femtosecond laser at different single-pulse energies. Experimental results show that the transformation thresholds of the Si surface and the C surface are 5.60 J/cm2 and 6.40 J/cm2, corresponding to the modification thresholds of 2.26 J/cm2 and 2.42 J/cm2, respectively. The Raman and EDS results reveal that there are no phase transformations or material changes on different surfaces of 6H-SiC at low energy, however, decomposition and oxidation occur and then accumulate into dense new phase material under high-energy laser irradiation. We found that the distribution of structural phase transformation is uneven from the center of the spot to the edge. The content of this research reveals the internal evolution mechanism of high-quality laser processing of hard material 6H-SiC. We expect that this research will contribute to the further development of SiC-based MEMS devices.

Peptide S4 is an entry inhibitor of SARS-CoV-2 infection.

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a significant socioeconomic burden, and combating COVID-19 is imperative. Blocking the SARS-CoV-2 RBD-ACE2 interaction is a promising therapeutic approach for viral infections, as SARS-CoV-2 binds to the ACE2 receptors of host cells via the RBD of spike proteins to infiltrate these cells. We used computer-aided drug design technology and cellular experiments to screen for peptide S4 with high affinity and specificity for the human ACE2 receptor through structural analysis of SARS-CoV-2 and ACE2 interactions. Cellular experiments revealed that peptide S4 effectively inhibited SARS-CoV-2 and HCoV-NL63 viruses from infecting host cells and was safe for cells at effective concentrations. Based on these findings, peptide S4 may be a potential pharmaceutical agent for clinical application in the treatment of the ongoing SARS-CoV-2 pandemic.

[Spatial structure and geographic distribution of major active component in medical herb Dipsacus asperoides population].

To study the spatial structure and geographic distribution of major active components in medical herb Dipsacus asperoides population, in Guizhou province, the author obtained the asperosaponin VI content among 38 D. asperoides population points. Using spatial statistic and traditional statistic, the spatial variation pattern has been found. Combining the environment variables, it is tried to reveal the geographic distribution rules between group of major active component in medical herb D. asperoides population and group of environmental factors. The results show: the spatial structure of major active component in medical herb D. asperoides population is randomly distributed in global space, but has two aggregation points which have a high positive correlation in local space; environmental factors value for group of major active component in medical herb D. asperoides population has a large difference in the interior-group, but environmental factors value has a small difference in inter-group.

[Application of hyperspectral remote sensing in field of medicinal plants monitoring research].

The paper introduces the basic concept and characteristics of hyperspectral remote sensing, and analyzed the application of hyperspectral remote sensing in the field of plants research. On the basis of the research advances of hyperspectral plant study, paper also analyzed the key facts that effects the application of hyperspectral remote sensing on the some researches which include distinguishing species,monitoring growth and quality etc. It proposed a new ideas and methods for people to research medicinal plants.

[Data validation methods and discussion on Chinese materia medica resource survey].

From the beginning of the fourth national survey of the Chinese materia medica resources, there were 22 provinces have conducted pilots. The survey teams have reported immense data, it put forward the very high request to the database system construction. In order to ensure the quality, it is necessary to check and validate the data in database system. Data validation is important methods to ensure the validity, integrity and accuracy of census data. This paper comprehensively introduce the data validation system of the fourth national survey of the Chinese materia medica resources database system, and further improve the design idea and programs of data validation. The purpose of this study is to promote the survey work smoothly.

[Resources investigation of medicinal plants of Huangfu mountain in city of Chuzhou based on grid sampling method].

OBJECTIVE: To conduct preliminary investigation to the species and reserves of medicinal plants in Huangfu Mountain, and to provide references to the general survey of those plants for medicine. METHOD: Combined with global positioning system (GPS), the program of investigation with grid sampling was used in this resource survey of medicinal plants. RESULT: After the preliminary investigation of the plants for medical use of Huangfu Mountain, it is found that there are 103 families with 313 kinds of plants. There are many medicinal plants and large distribution, such as Pseudostellaria heterophylla, Semiaguilegia adoxoides and Pinellia ternate. CONCLUSION: Huangfu Mount, with so many different kinds of medicinal plants and comfortable environment for part of the medicinal plants to grow, could be developed as a base for planting Chinese herbal medicines.

[Research on reserves estimation method of wild medicinal plants resource for generous species based on spatial information technology].

Traditional Chinese medicinal resource survey method is time-consuming, strenuous, and having great human influence, the precision is not high enough. This paper, by using spatial information technology, carries on spatial sampling survey for wild medicinal plants resource for generous species to arrange the quadrat scientifically and estimate the suitable area, reserve precisely of medicinal plants. It not only improves the survey precision, but reduces the workload of field survey and provides scientific basis for survey method of pilot work on the fourth national traditional Chinese medicinal resource census.

EEG emotion recognition using improved graph neural network with channel selection.

BACKGROUND AND OBJECTIVE: Emotion classification tasks based on electroencephalography (EEG) are an essential part of artificial intelligence, with promising applications in healthcare areas such as autism research and emotion detection in pregnant women. However, the complex data acquisition environment provides a variable number of EEG channels, which interferes with the model to simulate the process of information transfer in the human brain. Therefore, this paper proposes an improved graph convolution model with dynamic channel selection. METHODS: The proposed model combines the advantages of 1D convolution and graph convolution to capture the intra- and inter-channel EEG features, respectively. We add functional connectivity in the graph structure that helps to simulate the relationship between brain regions further. In addition, an adjustable scale of channel selection can be performed based on the attention distribution in the graph structure. RESULTS: We conducted various experiments on the DEAP-Twente, DEAP-Geneva, and SEED datasets and achieved average accuracies of 90.74%, 91%, and 90.22%, respectively, which exceeded most existing models. Meanwhile, with only 20% of the EEG channels retained, the models achieved average accuracies of 82.78%, 84%, and 83.93% on the above three datasets, respectively. CONCLUSIONS: The experimental results show that the proposed model can achieve effective emotion classification in complex dataset environments. Also, the proposed channel selection method is informative for reducing the cost of affective computing.