Original Line Research Articles

Analysis of Dynamic Capacity Increase Transmission Line Carrying Capacity under Temperature and Electromagnetic Field

This article mainly introduces that, as the proportion of renewable energy to total consumed energy increases further, with the large-scale development and construction of new energy, the local power grid may face the problem of conflicting consumption in the future, and the original transmission lines will no longer meet the power demand. For this reason, dynamic capacity increase technology is used to calculate the load capacity in real-time. Through simulation analysis under temperature and electromagnetic fields to ensure that the conductor has sufficient safety while increasing the load capacity, clear data results are obtained based on the analysis.

A Novel 2-D Slotted Structure Extended Interaction Oscillator

Numerous vacuum electronic devices (VEDs) have been widely studied, among which extended interaction oscillators (EIO) exhibit appealing characteristics, such as high frequency, high efficiency, lightweight, low working voltage, and stable working frequency. The slow wave structure typically adopts the coupled-cavity, folded-waveguide, ladder line, and other structures. The ladder line has a simple structure and is easy to process, with desirable heat dissipation and large power capacity. In this study, a novel slow wave structure being longitudinally slotted in the transverse grating of the original ladder line is proposed, and the 2-D slotted structure can be obtained. Encouragingly, the structure not only increases the cross-sectional area of the electron beam but also, more importantly, increases the effective interaction area to considerably improve the output power. The dispersion characteristics and field distribution are first studied through numerical and simulation calculations, and the optimal slotted structure parameters and working parameters are analyzed using PIC software. Next, a 94-GHz 2-D slotted structure EIO is designed. Finally, compared with the ladder line, the cavity characteristic impedance <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\textit{R}$</tex-math> </inline-formula> / <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\textit{Q}$</tex-math> </inline-formula> is increased by 52.5%, and the output power is increased by 21.1%. This novel structure is expected to provide a new idea for the power enhancement of millimeter-wave and terahertz EIOs.

An Improved Crack Breathing Model and Its Application in Crack Identification for Rotors

The crack breathing model and crack identification method for rotors using nonlinearity induced by cracks are studied in this work. Firstly, the finite element method is utilized to model a rotor–bearing system with a response-dependent breathing crack to obtain the numerical data for crack identification. During the modelling, an improved breathing crack model is proposed, focused on the unreasonable assumption about crack closure line in the original crack closure line position (CCLP) model. Compared with the original model, the improved breathing model can reflect the nonlinear behavior of cracks better. Secondly, based on the established model, super-harmonic features at 1/3 and 1/2 of the critical rotating speeds under different crack locations and crack depths are extracted for crack identification. Additionally, the super-harmonic features from two measurement points are used as inputs into an artificial neural network with a Levenberg–Marquardt back-propagation algorithm, corresponding crack positions and depths as outputs. The robustness of the method is tested by examining the identification results under different levels of noise. The results show that the proposed crack identification method is efficient for simultaneous identification of crack depth and position in operating rotors.

Measurement and numerical analysis on dynamic performance of the LMS maglev train-track-continuous girder coupled system with running speed-up state

Improving the target running speed on original maglev lines is an economical method to meet the social development demands. With the speed improvement, maglev train-track-bridge system coupled vibration is intensified, seriously affecting the train’s moving stability. For this study, vibration performance about the coupled system at measured speeds (100 km/h–150 km/h) are investigated based on field test; then a refined numerical coupling vibration model is developed and validated; finally, vibration characteristics at target speed of 160 km/h are studied by simulation. Results show the accelerations of maglev vehicle increase significantly when running speeds exceed 140 km/h, and F-rail’s acceleration is noticeably larger than that of continuous girder and sleeper. The results of this numerical model agree well with those of the field test. When the speed reaches the target of 160 km/h, the amplitude increases in time domain, and dynamic responses are within a reasonable range and meet the standard requirements.

Establishment and characterization of a novel cancer stem-like cell of cholangiocarcinoma.

Cholangiocarcinoma (CCA) is an aggressive malignant tumor of bile duct epithelia. Recent evidence suggests the impact of cancer stem cells (CSC) on the therapeutic resistance of CCA; however, the knowledge of CSC in CCA is limited due to the lack of a CSC model. In this study, we successfully established a stable sphere-forming CCA stem-like cell, KKU-055-CSC, from the original CCA cell line, KKU-055. The KKU-055-CSC exhibits CSC characteristics, including: (1) the ability to grow stably and withstand continuous passage for a long period of culture in the stem cell medium, (2) high expression of stem cell markers, (3) low responsiveness to standard chemotherapy drugs, (4) multilineage differentiation, and (5) faster and constant expansive tumor formation in xenograft mouse models. To identify the CCA-CSC-associated pathway, we have undertaken a global proteomics and functional cluster/network analysis. Proteomics identified the 5925 proteins in total, and the significantly upregulated proteins in CSC compared with FCS-induced differentiated CSC and its parental cells were extracted. Network analysis revealed that high mobility group A1 (HMGA1) and Aurora A signaling through the signal transducer and activator of transcription 3 pathways were enriched in KKU-055-CSC. Knockdown of HMGA1 in KKU-055-CSC suppressed the expression of stem cell markers, induced the differentiation followed by cell proliferation, and enhanced sensitivity to chemotherapy drugs including Aurora A inhibitors. In silico analysis indicated that the expression of HMGA1 was correlated with Aurora A expressions and poor survival of CCA patients. In conclusion, we have established a unique CCA stem-like cell model and identified the HMGA1-Aurora A signaling as an important pathway for CSC-CCA.

Improvement of CNR in low-count PET scans using tissue-scattered data as initial estimate in non-TOF MLEM reconstruction

We have proposed a method to improve contrast-to-noise ratios (CNRs) of lesions in low-count PET scans using the rejected single scattered (inside tissue) data. Indeed, with the advent of high-quality detectors regarding their energy and time resolution, and list mode data acquisition, adding single scattered events by estimating their original line of response has become more and more feasible. The low-count PET scan data were simulated using GATE software considering Jaszczak type object, and images were reconstructed using STIR. A deconvolution based method to create images from rejected single scatter data has been proposed. These scatter images were, in turn, used as an initial estimate in non-TOF MLEM reconstruction. Three types of human-sized PET scanners—ideal, state-of-art, and future generation—with different timing and energy resolutions were considered. We found a significant improvement in trade-off between the contrast recovery coefficient (CRC) and coefficient of variation (CoV) for the non-TOF MLEM reconstructed images while comparing with other less computational ways of creating initial estimates. CNR improvements were found for all lesions. The present work demonstrated the beneficial use of tissue-scattered TOF PET events which constitutes a high percentage of data in PET imaging, and rejected in general. Our approach has a potential and scope for further study.

Case Report on Painful Patellar Crepitation Following a Knee Replacement with Preserved Patella

Patellar clunk crepitation is a well-known complication following knee replacement surgery and is associated with posterior stabilized knee replacement surgery and surgical technique. Currently, patellar clunk or crepitation management following knee replacement surgery with preserved patella remains unclear. The purpose of this case report is to discuss whether patellar clunk or crepitation management should include debridement with patellar resurfacing or debridement alone. This case describes a patellar crepitation after knee replacement surgery with the preserved patella. The surgery went uneventfully using the standard medial parapatellar approach. However, the patient was still unsatisfied with the chronic left knee pain (&gt;3 months) and crepitation that developed following the surgery, and the patient was diagnosed with patellar clunk and crepitation (PCC). A patellar resurfacing procedure was performed with a satisfactory clinical outcome. Replicating the original joint line level and placing the tibial component posteriorly play a pivotal role in preventing PCC. Debridement and patellar resurfacing procedures are recommended in this type of case to overcome the valgus knee alignment and the placement of the tibial component.

ON PRODUCTION OF INTERLINE HYBRIDS OF GEESE

Работа выполнена в племенном заводе ООО «Вурнарец», Республика Чувашия. Исходным материалом для селекции служила линдовская порода гусей. В результате 11 поколений дифференцированной селекции гусей по продуктивным признакам, по отцовской линии была увеличена живая масса в 9-недельном возрасте на 29,3%, затраты корма снижены на 19,6%. За этот период по материнской линии выход гусят от несушки был увеличен на 18,8% соответственно. При скрещивании гусаков отцовской линии с гусынями материнской комплексный показатель (выход мяса от несушки в живой массе) был выше, чем в исходных линиях, на 10,0−17,2%. The work was carried out in the breeding plant OOO "Vurnarets" Republic of Chuvashia. The source material for selection was the Lindovskaya breed of geese. As a result of 11 generations of differentiated selection of geese by productive characteristics, on the paternal line the live weight at 9-week age was increased by 29.3%, feed costs were reduced by 19.6%. During this period on the maternal side, the yield of the goslings from the laying bird was increased by 18.8%, respectively. When crossing the goose-gander of the paternal line with the maternal geese the complex indicator (meat yield from the laying bird in live weight) was higher than in the original lines by 10.0−17.2%.

Dehydrin AmDH1 improves drought tolerance in transgenic maize plants

Dehydrin is a large protein group to have protective functions to plant cells under osmotic stress. Overexpression of dehydrin encoded genes isolated from mangrove plants had improved drought tolerance of transgenic plants. Therefore, in this research, a coding sequence with the length of 573bp that encodes Dehydrin1 (DH1) protein was isolated from a mangrove plant namely Mam bien (Avicennia marina) which naturally grows in Xuan Thuy national park, Vietnam. The coding sequence was denoted as AmDH1 and cloned into the plant transformation vector pCAM 35S-AmDH1. The transgenic vector was transformed into A. tumefiaciens cells and the vector was transferred into the K7 maize line to generate transgenic maize seedlings. The AmDH1 transgenic maize T2 seedlings have shown an improved drought tolerance than the original K7 maize line.

Electrical design for the conversion of existing UK HVAC overhead lines to HVDC operation under severe pollution conditions, ranging from 170 to 500 kV DC

This paper provides the HVDC electrical design of three existing UK HVAC overhead lines, two of which constitute 85% of the UK transmission overhead line system. The designs are based on CIGRE and IEC recommendations. Also, the verification of the electrical stress requirements on the insulators is performed by means of finite element simulations. The paper shows that the HVDC converted tower lines would carry 1.6 to 2.9 times the power of the original HVAC lines, depending on tower type, insulation arrangement, and environmental conditions.

The Mechanisms of GPR55 Receptor Functional Selectivity during Apoptosis and Proliferation Regulation in Cancer Cells

GPR55 is a non-canonical cannabinoid receptor, important for cancer proliferation. Depending on the ligand, it induces either cell proliferation or death. The objective of the study was to establish the mechanisms of this multidirectional signaling. Using the CRISPR-Cas9 system, the GPR55, CB1, CB2, and GPR18 receptor knockouts of the MDA-MB-231 line were obtained. After the CB2 receptor knockout, the pro-apoptotic activity of the pro-apoptotic ligand docosahexaenoyl dopamine (DHA-DA) slightly increased, while the pro-proliferative activity of the most active synthetic ligand of the GPR55 receptor (ML-184) completely disappeared. On the original cell line, the stimulatory effect of ML-184 was removed by the CB2 receptor blocker and by GPR55 receptor knockout. Thus, it can be confidently assumed that when proliferation is stimulated with the participation of the GPR55 receptor, a signal is transmitted from the CB2 receptor to the GPR55 receptor due to the formation of a heterodimer. GPR18 was additionally involved in the implementation of the pro-apoptotic effect of DHA-DA, while the CB1 receptor is not involved. In the implementation of the pro-apoptotic action of DHA-DA, the elimination of Gα13 led to a decrease in cytotoxicity. The obtained data provide novel details to the mechanism of the pro-proliferative action of GPR55.

Abstract P1-13-20: Reprogramming of oncogenic signal pathways and metabolism during trastuzumab resistance formation of HER2 positive breast cancer

Abstract Background The formation of resistance against trastuzumab deeply affects the treatment of HER2 positive breast cancer. Although studies have demonstrated several possible reasons that cause the trastuzumab resistance, the precise changes of cell process as well as the interaction between cancer cells and tumor microenvironment during the formation of resistance are still poorly understood. Here we figured out several crucial changes including oncogenic signal pathways as well as metabolism processes especially amino acids and polyunsaturated fatty acids (PUFAs) during the establishment of trastuzumab resistance. We further suggested that the inducers of ferroptosis may be promising reagents for trastuzumab resistant HER2 positive breast cancer. Methods The trastuzumab resistant cell was generated from a sensitive cell line SKBR3. To simulate the adaptation process of HER2 positive tumor to anti-tumor drug, the concentration of trastuzumab was changed gently to prevent cell death. After 8 weeks treatment, trastuzumab concentration in medium was raised from 1μg/ml to 6μg/ml, and a “persister” cell line SKBR3_HP was obtained. The original cell line SKBR3 was cultured for 8 weeks as well to exclude any changes that caused by culture condition. And each cell line includes 3 biological repeats. The viability of two cell lines is measured by CCK8 cell proliferation test with different concentration of trastuzumab. Total RNA of both SKBR3 and SKBR3_HP cells were prepared for sequencing. Activity scores of oncogenic signal pathways as well as metabolism processes were defined and calculated as the relative gene expression value averaged over all genes in this pathway in certain cell type. Flow cytometry was applied for reactive oxygen species measurement with BODIPY-C11. Results After 8 weeks treatment of trastuzumab, the persister cell line SKBR3_HP showed a significant higher viability than original sensitive cell line SKBR3. Oncogenic signal pathway analysis revealed that RTK-Ras, MYC and HIPPO pathways turn into a more active state in SKBR3_HP cells, as well as the upregulation of several cell cycle genes, which are all the response to the blockage of HER2 signal cascade and together maintains cell proliferation. Beside signal pathway, the reprogramming of metabolism is also the consequence of cell adaptation to trastuzumab. Minor changes in energy metabolism, such as glycolysis, citrate cycle and oxidative phosphorylation were observed in SKBR3_HP cells. However, amino acid metabolism, including the synthesis of phenylalanine, histidine, arginine, proline, cysteine and methionine, was largely enhanced in SKBR3_HP cells. Fatty acid, specifically the metabolism of PUFAs, such as linoleic acid, alpha-linolenic acid and arachidonic acid, was activated during the formation of trastuzumab resistance, which was confirmed by the lipid metabolomics data that most n-3/n-6 PUFAs decreased in SKBR3_HP cells. As cysteine and PUFAs metabolism might closely associate with cellular redox balances, erastin and RSL3 were applied to interrupt the intake of cysteine and potentiate the lipid peroxidation process, respectively. A situation of higher ferroptosis sensitivity accompanies raising peroxidated lipids could be detected in SKBR3_HP cells. Conclusion By analyzing the transcriptome and metabolomics data of trastuzumab sensitive and persister cell lines, we pointed out that the changes of oncogenic signal pathway, together with metabolism variation, particularly amino acids and PUFAs, are all the cellular adaptations to high trastuzumab environment. And the higher ferroptosis sensitivity of persister cells could be a valuable treatment target. Citation Format: Ningjun Duan, Yijia Hua, Shuang Hu, Yongmei Yin. Reprogramming of oncogenic signal pathways and metabolism during trastuzumab resistance formation of HER2 positive breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P1-13-20.

Deep convolutional neural network for partial discharge monitoring system

In the insulating material, the event of PDs takes place due to the air pockets, voids, or imperfections in such fields. In HV components like cables, the PD detection aids in ascertaining the insulating material condition. Consequently, the territory of PD estimation and conclusion is acknowledged as one of the most significant non-destructive methods for surveying the quality and specialized respectability of HV power mechanical assembly and links. In this work, a new data-driven model to recognize the condition of PD pulses of power cables is proposed with the aid of optimized Convolutional Neural Networks (CNNs). The two main stages of the developed framework are feature extraction and recognition. The data downloaded by VSB from the power cable is subjected to dimensionality reduction via Principal Component Analysis (PCA). Then, from these signals, the features are extracted with the aid of technical indicators like Rate of change (ROC), Relative Strength Index (RSI), Adaptive Moving Average (AMA), and Standard Deviation. Additionally, the features of the original power line data are also extracted. These features are put into an enhanced CNN as input. A new hybrid version known as the Combined Sealion-Swarm Optimization Algorithm (CS-SOA) optimizes the weight and activating function of CNN to increase the accuracy of the PD condition observation. From the result, it can be noticed that the accuracy of the proposed model, is 77.7%, 5.55%, and 3.335% higher than the existing models like SVM, CNN, and LSTM at TP= 80.

Subsoiling for planting trees in dehesa system: long-term effects on soil organic carbon

Incorporating trees into agricultural systems, including grasslands, increases the soil organic carbon sequestration and contributes to climate change mitigation. Site preparation for tree establishment is a common practice that can involve a variety of techniques and agricultural implements such as subsoiling. This study aimed to evaluate the long-term effects of subsoiling on soil organic carbon (SOC) concentrations and stocks in a Mediterranean grassland afforested with holm oaks 22 years ago and now converted into a Dehesa agroforestry system. The study was conducted in a dehesa farm in Southwest Spain. Soil samples were taken at six depths under 10 tree canopies within and outside the original subsoiling line. Subsoiling significantly decreased SOC concentration. Mean SOC concentration in the first 20 cm was 30% lower at the subsoiling line. SOC stocks for the first 60 cm were 2660 g m−2 within the subsoiling line and 4320 g m−2 outside the line. There was a clear reduction in SOC concentration and stock with increasing depth. Root abundance and deeper rooting increased with subsoiling but did not translate into sufficient carbon accumulation in the soil, which is moderate even after 22 years. This study reveals that, in the long term, there is a trade-off in CO2 sequestration between tree planting and soil subsoiling, highlighting the need for further research into the potential benefits and detriments of subsoiling.

CRISPR-gene-engineered CYBB knock-out PLB-985 cells, a useful model to study functional impact of X-linked chronic granulomatous disease mutations: application to the G412E X91+-CGD mutation.

Chronic granulomatous disease (CGD) is a rare primary immune disorder caused by mutations in one of the five subunits of the NADPH oxidase complex expressed in phagocytes. Two-thirds of CGD cases are caused by mutations in CYBB that encodes NOX2 or gp91phox. Some rare X91+-CGD point mutations lead to a loss of function but with a normal expression of the mutated NOX2 protein. It is therefore necessary to ensure that this mutation is indeed responsible for the loss of activity in order to make a safe diagnosis for genetic counselling. We previously used the X-CGD PLB-985 cell model of M.C. Dinauer obtained by homologous recombination in the original PLB-985 human myeloid cell line, in order to study the functional impact of such mutations. Although the PLB-985 cell line was originally described by K.A. Tucker et al. in1987 as a distinct cell line isolated from a patient with acute nonlymphocytic leukemia, it is actually identified as a subclone of the HL-60 cells. In order to use a cellular model that meets the quality standard for the functional study of X91+-CGD mutations in CGD diagnosis, we developed our own model using the CRISPR-Cas9 technology in a certified PLB-985 cell line from DSMZ-German Collection of Microorganisms and Cell Cultures. Thanks to this new X-CGD model, we demonstrated that the G412E mutation in NOX2 found in a X91+-CGD patient prohibits access of the electron donor NADPH to its binding site explaining the absence of superoxide production in his neutrophils.

Production and Characterization of K562 Cellular Clones Hyper-Expressing the Gene Encoding α-Globin: Preliminary Analysis of Biomarkers Associated with Autophagy.

One of the most relevant pathophysiological hallmarks of β-thalassemia is the accumulation of toxic α-globin chains inside erythroid cells, which is responsible for their premature death (hemolysis). In this context, the availability of an experimental model system mimicking the excess in α-globin chain production is still lacking. The objective of the present study was to produce and characterize K562 cellular clones forced to produce high amounts of α-globin, in order to develop an experimental model system suitable for studies aimed at the reduction of the accumulation of toxic α-globin aggregates. In the present study, we produced and characterized K562 cellular clones that, unlike the original K562 cell line, stably produced high levels of α-globin protein. As expected, the obtained clones had a tendency to undergo apoptosis that was proportional to the accumulation of α-globin, confirming the pivotal role of α-globin accumulation in damaging erythroid cells. Interestingly, the obtained clones seemed to trigger autophagy spontaneously, probably to overcome the accumulation/toxicity of the α-globin. We propose this new model system for the screening of pharmacological agents able to activate the full program of autophagy to reduce α-globin accumulation, but the model may be also suitable for new therapeutical approaches targeted at the reduction of the expression of the α-globin gene.

Engineering protein glycosylation in CHO cells to be highly similar to murine host cells.

Since 2015 more than 34 biosimilars have been approved by the FDA. This new era of biosimilar competition has stimulated renewed technology development focused on therapeutic protein or biologic manufacturing. One challenge in biosimilar development is the genetic differences in the host cell lines used to manufacture the biologics. For example, many biologics approved between 1994 and 2011 were expressed in murine NS0 and SP2/0 cell lines. Chinese Hamster ovary (CHO) cells, however, have since become the preferred hosts for production due to their increased productivity, ease of use, and stability. Differences between murine and hamster glycosylation have been identified in biologics produced using murine and CHO cells. In the case of monoclonal antibodies (mAbs), glycan structure can significantly affect critical antibody effector function, binding activity, stability, efficacy, and in vivo half-life. In an attempt to leverage the intrinsic advantages of the CHO expression system and match the reference biologic murine glycosylation, we engineered a CHO cell expressing an antibody that was originally produced in a murine cell line to produce murine-like glycans. Specifically, we overexpressed cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) and N-acetyllactosaminide alpha-1,3-galactosyltransferase (GGTA) to obtain glycans with N-glycolylneuraminic acid (Neu5Gc) and galactose-α-1,3-galactose (alpha gal). The resulting CHO cells were shown to produce mAbs with murine glycans, and they were then analyzed by the spectrum of analytical methods typically used to demonstrate analytical similarity as a part of demonstrating biosimilarity. This included high-resolution mass spectrometry, biochemical, as well as cell-based assays. Through selection and optimization in fed-batch cultures, two CHO cell clones were identified with similar growth and productivity criteria to the original cell line. They maintained stable production for 65 population doubling times while matching the glycosylation profile and function of the reference product expressed in murine cells. This study demonstrates the feasibility of engineering CHO cells to express mAbs with murine glycans to facilitate the development of biosimilars that are highly similar to marketed reference products expressed in murine cells. Furthermore, this technology can potentially reduce the residual uncertainty regarding biosimilarity, resulting in a higher probability of regulatory approval and potentially reduced costs and time in development.

A new mouse model of elastin haploinsufficiency highlights the importance of elastin to vascular development and blood pressure regulation

Supravalvular aortic stenosis (SVAS) is an autosomal dominant disease resulting from elastin (ELN) haploinsufficiency. Individuals with SVAS typically develop a thickened arterial media with an increased number of elastic lamellae and smooth muscle cell (SMC) layers and stenosis superior to the aortic valve. A mouse model of SVAS (Eln+/−) was generated that recapitulates many aspects of the human disease, including increased medial SMC layers and elastic lamellae, large artery stiffness, and hypertension. The vascular changes in these mice were thought to be responsible for the hypertension phenotype. However, a renin gene (Ren) duplication in the original 129/Sv genetic background and carried through numerous strain backcrosses raised the possibility of renin-mediated effects on blood pressure. To exclude excess renin activity as a disease modifier, we utilized the Cre-LoxP system to rederive Eln hemizygous mice on a pure C57BL/6 background (Sox2-Cre;Elnf/f). Here we show that Sox2-Cre;Eln+/f mice, with a single Ren1 gene and normal renin levels, phenocopy the original global knockout line. Characteristic traits include an increased number of elastic lamellae and SMC layers, stiff elastic arteries, and systolic hypertension with widened pulse pressure. Importantly, small resistance arteries of Sox2-Cre;Eln+/f mice exhibit a significant change in endothelial cell function and hypercontractility to angiotensin II, findings that point to pathway-specific alterations in resistance arteries that contribute to the hypertensive phenotype. These data confirm that the cardiovascular changes, particularly systolic hypertension, seen in Eln+/− mice are due to Eln hemizygosity rather than Ren duplication.

Generation of a TRPM8 knockout hESC line (WAe009-A-A) derived from H9 using CRISPR/Cas9

The transient receptor potential cation channel subfamily M member 8 (TRPM8) is a kind of non-selective cation channel which controls Ca2+ homeostasis. Mutations in TRPM8 were related to dry eye diseases (DED). Here we constructed a TRPM8 knockout cell line WAe009-A-A from the original embryonic stem cell line H9 using CRISPR/Cas9 technology, which maybe helpful for exploring the pathogenesis of DED. WAe009-A-A cells possess stem cell morphology and pluripotency as well as normal karyotype, and have the ability of differentiating into three germ layers in vitro.

Elimination of the yellow pigment gene PSY-E2 tightly linked to the Fusarium head blight resistance gene Fhb7 from Thinopyrum ponticum

Fhb7 is a major gene that was transferred from Thinopyrum ponticum to chromosome 7D of wheat (Triticum aestivum) and confers resistance to both Fusarium head blight (FHB) and Fusarium crown rot (FCR). However, Fhb7 is tightly linked to the PSY-E2 gene, which causes yellow flour, limiting its application in breeding. To break this linkage, marker K-PSY was developed for tagging PSY-E2 and used with Fhb7 markers to identify recombination between the two genes. Screening 21,000 BC1F2 backcross progeny (Chinese Spring ph1bph1b*2/SDAU 2028) revealed two Fhb7+ wheat-Tp7el2L lines, Shannong 2–16 and Shannong 16–1, that carry a desired truncated Fhb7+ translocation segment without PSY-E2. The two lines show levels of resistance to FHB and FCR similar to those of the original translocation line SDAU 2028, but have white flour. To facilitate Fhb7 use in wheat breeding, STS markers were developed and used to isolate Fhb7 on a truncated Tp7el2 translocation segment. Near-isogenic lines carrying the Fhb7+ segment were generated in the backgrounds of three commercial cultivars, and Fhb7+ lines showed increased FHB and FCR resistance without yield penalty. The breakage of the tight linkage between Fhb7 and PSY-E2 via homoeologous recombination provides genetic resources for improvement of wheat resistance to FHB and FCR and permit the large-scale deployment of Fhb7 in breeding using marker-assisted selection.