DNA methylation patterns in the peripheral blood of Xinjiang brown cattle with variable somatic cell counts.

The use of wide-ranging dairy herd improvement (DHI) measurements has resulted in the investigation of somatic cell count (SCC) and the identification of many genes associated with mastitis resistance. In this study, blood samples of Xinjiang brown cattle with different SCCs were collected, and genome-wide DNA methylation was analyzed by MeDIP-seq. The results showed that peaks were mostly in intergenic regions, followed by introns, exons, and promoters. A total of 1,934 differentially expressed genes (DEGs) associated with mastitis resistance in Xinjiang brown cattle were identified. The enrichment of differentially methylated CpG islands of the TRAPPC9 and CD4 genes was analyzed by bisulfate genome sequencing. The methylation rate of differentially methylated CpGs was higher in the TRAPPC9 gene of cattle with clinical mastitis (mastitis group) compared with healthy cattle (control group), while methylation of differentially methylated CpGs was significantly lower in CD4 of the mastitis group compared with the control group. RT-qRCR analysis showed that the mastitis group had significantly reduced expression of CD4 and TRAPPC9 genes compared to the control group (p < 0.05). Furthermore, Mac-T cells treated with lipopolysaccharide and lipoteichoic acid showed significant downregulation of the TRAPPC9 gene in the mastitis group compared with the control group. The identified epigenetic biomarkers provide theoretical reference for treating cow mastitis, breeding management, and the genetic improvement of mastitis resistance in Xinjiang brown cattle.

Comparative Study of Bacterial Microbiota Differences in the Rumen and Feces of Xinjiang Brown and Holstein Cattle.

Xinjiang Brown cattle are a unique and widely distributed breed of dual-purpose cattle in the Xinjiang area of China, whose milk production performance differs from Holstein cattle. It has been known that variations in bacterial species of the gastrointestinal tract influence milk protein, fat, and lactose synthesis. However, the microbiota differences between Xinjiang Brown and Holstein cattle are less known. This study aims to compare the bacterial community composition of the rumen and feces of these two cattle breeds under the same dietary and management conditions. The 16s rRNA sequencing data and milk production of 18 Xinjiang Brown cows and 20 Holstein cows on the same farm were obtained for analysis. The results confirmed differences in milk production between Xinjiang Brown and Holstein cattle. Microbiota with different relative abundance between these two cattle breeds were identified, and their biological functions might be related to milk synthesis. This study increases the understanding of the differences in microbiota between Xinjiang Brown and Holstein cattle and might provide helpful information for microbiota composition optimization of these dairy cattle.

Estimation of Genetic Parameters for Early Growth Traits in Luzhong Mutton Sheep.

In this study, six different animal models were fitted, and the constrained maximum likelihood method was used to assess the genetic parameters and genetic trends of early growth traits in Luzhong mutton sheep. The experimental data of this study included the newborn weight (BWT, N = 2464), weaning weight (WWT, N = 2923), weight at 6 months of age (6WT, N = 2428), average daily weight gain from birth to weaning (ADG1, N = 2424), and average daily weight gain from weaning to 6 months of age (ADG2, N = 1836) in Luzhong mutton sheep (2015~2019). The best model for the genetic parameters of the five traits in Luzhong mutton sheep was identified as Model 4 using the Akaike information criterion (AIC) and likelihood ratio test (LRT) methods, in which the estimated values of direct heritability for the BWT, WWT, 6WT, ADG1, and ADG2 were 0.156 ± 0.057, 0.547 ± 0.031, 0.653 ± 0.031, 0.531 ± 0.035, and 0.052 ± 0.046, respectively, and the values for maternal heritability were 0.201 ± 0.100, 0.280 ± 0.047, 0.197 ± 0.053, 0.275 ± 0.052, and 0.081 ± 0.092, respectively. The genetic correlation between the ADG2 and WWT was negative, and the genetic and phenotypic correlations among the remaining traits were positive. In this study, maternal effects had a more significant influence on early growth traits in Luzhong mutton sheep. In conclusion, to effectively improve the accuracy of genetic parameter estimation, maternal effects must be fully considered to ensure more accurate and better breeding planning.

Primulaweiliei (Primulaceae), a new species from Hubei, Central China.

In this study, we describe and illustrate a new species, Primulaweiliei L.S.Yang, Z.K.Wu & G.W.Hu, from the Shennongjia Forestry District, Hubei Province in Central China. It is morphologically assigned to Primulasect.Aleuritia based on its dwarf and hairless habit, long petiole, fruits longer than calyx and covered by farina on the scape. This new species is similar to P.gemmifera and P.munroisubsp.yargongensis in the same section, but it can be distinguished by its smaller calyxes, homostylous flowers, corolla tube throat without annular appendage and only 1-2 flowers in each inflorescence. Based on the assessment conducted according to the IUCN Red List criteria, we propose that P.weiliei be classified as a Critically Endangered (CR) species.

Genomic prediction based on a joint reference population for the Xinjiang Brown cattle.

Introduction: Xinjiang Brown cattle constitute the largest breed of cattle in Xinjiang. Therefore, it is crucial to establish a genomic evaluation system, especially for those with low levels of breed improvement. Methods: This study aimed to establish a cross breed joint reference population by analyzing the genetic structure of 485 Xinjiang Brown cattle and 2,633 Chinese Holstein cattle (Illumina GeneSeek GGP bovine 150 K chip). The Bayes method single-step genome-wide best linear unbiased prediction was used to conduct a genomic evaluation of the joint reference population for the milk traits of Xinjiang Brown cattle. The reference population of Chinese Holstein cattle was randomly divided into groups to construct the joint reference population. By comparing the prediction accuracy, estimation bias, and inflation coefficient of the validation population, the optimal number of joint reference populations was determined. Results and Discussion: The results indicated a distinct genetic structure difference between the two breeds of adult cows, and both breeds should be considered when constructing multi-breed joint reference and validation populations. The reliability range of genome prediction of milk traits in the joint reference population was 0.142-0.465. Initially, it was determined that the inclusion of 600 and 900 Chinese Holstein cattle in the joint reference population positively impacted the genomic prediction of Xinjiang Brown cattle to certain extent. It was feasible to incorporate the Chinese Holstein into Xinjiang Brown cattle population to form a joint reference population for multi-breed genomic evaluation. However, for different Xinjiang Brown cattle populations, a fixed number of Chinese Holstein cattle cannot be directly added during multi-breed genomic selection. Pre-evaluation analysis based on the genetic structure, kinship, and other factors of the current population is required to ensure the authenticity and reliability of genomic predictions and improve estimation accuracy.

Whole-genome resequencing uncovers diversity and selective sweep in Kazakh cattle.

The Kazakh cattle in the Xinjiang Uygur Autonomous Region of China are highly adaptable and have multiple uses, including milk and meat production, and use as draft animals. They are an excellent original breed that could be enhanced by breeding and hybrid improvement. However, the genomic diversity and signature of selection underlying the germplasm characteristics require further elucidation. Herein, we evaluated 26 Kazakh cattle genomes in comparison with 103 genomes of seven other cattle breeds from regions around the world to assess the Kazakh cattle genetic variability. We revealed that the relatively low linkage disequilibrium at large SNP distances was strongly correlated with the largest effective population size among Kazakh cattle. Using population structural analysis, we next demonstrated a taurine lineage with restricted Bos indicus introgression among Kazakh cattle. Notably, we identified putative selected genes associated with resistance to disease and body size within Kazakh cattle. Together, our findings shed light on the evolutionary history and breeding profile of Kazakh cattle, as well as offering indispensable resources for germplasm resource conservation and crossbreeding program implementation.

Transcriptomic Changes and Regulatory Networks Associated with Resistance to Mastitis in Xinjiang Brown Cattle.

Xinjiang brown cattle are highly resistant to disease and tolerant of roughage feeding. The identification of genes regulating mastitis resistance in Xinjiang brown cattle is a novel means of genetic improvement. In this study, the blood levels of IL-1ß, IL-6, IL-10, TNF-α, and TGF-ß in Xinjiang brown cattle with high and low somatic cell counts (SCCs) were investigated, showing that cytokine levels were higher in cattle with high SCCs. The peripheral blood transcriptomic profiles of healthy and mastitis-affected cattle were constructed by RNA-seq. Differential expression analysis identified 1632 differentially expressed mRNAs (DE-mRNAs), 1757 differentially expressed lncRNAs (DE-lncRNAs), and 23 differentially expressed circRNAs (DE-circRNAs), which were found to be enriched in key pathways such as PI3K/Akt, focal adhesion, and ECM-receptor interactions. Finally, ceRNA interaction networks were constructed using the differentially expressed genes and ceRNAs. It was found that keynote genes or mRNAs were also enriched in pathways such as PI3K-Akt, cholinergic synapses, cell adhesion molecules, ion binding, cytokine receptor activity, and peptide receptor activity, suggesting that the key genes and ncRNAs in the network may play an important role in the regulation of bovine mastitis.

Strong interactions between solitons and background light in Brillouin-Kerr microcombs.

Dissipative Kerr-soliton combs are laser pulses regularly sustained by a localized solitary wave on top of a continuous-wave background inside a nonlinear resonator. Usually, the intrinsic interactions between the background light and solitons are weak and localized. Here, we demonstrate a strong interaction between the generated soliton comb and the background light in a Brillouin-Kerr microcomb system. This strong interaction enables the generation of a monostable single-soliton microcomb on a silicon chip. Also, new phenomena related to soliton physics including solitons hopping between different states as well as controlling the formations of the soliton states by the pump power, are observed owing to such strong interaction. Utilizing this monostable single-soliton microcomb, we achieve the 100% deterministic turnkey operation successfully without any feedback controls. Importantly, it allows to output turnkey ultra-low-noise microwave signals using a free-running pump.

Fault-tolerant controller design based on adaptive backstepping for tower cranes with actuator faults.

Due to the widespread application and significant investment required for a single crane, there is an increased emphasis on crane safety and service life. Fault-tolerant control as an effective solution to unexpected faults has been widely studied recently. However, most fault-tolerant control methods require redundant actuators or a complex design process, which is unsuitable for the tower crane. Following these problems, a fault-tolerant controller based on an adaptive backstepping technique is proposed. Firstly, the system states are reconstructed and written as a cascade system. Secondly, a fixed-time convergence optimized backstepping controller is proposed to achieve smooth control of the tower crane without generating sudden or abrupt values. Then, an adaptive approach has been proposed to update fault parameters for the crane system in case of a sudden fault occurrence. Finally, after conducting comparison tests, it has been determined that the proposed controller not only performs exceptionally well in terms of position accuracy and swing elimination, but also maintains a satisfactory control performance when faced with sudden faults.

Phylogenomics and divergence pattern of Polygonatum (Asparagaceae: Polygonateae) in the north temperate region.

Polygonatum is the largest genus of tribe Polygonateae (Asparagaceae) and is widely distributed in the temperate Northern Hemisphere, especially well diversified in southwestern China to northeastern Asia. Phylogenetic relationships of many species are still controversial. Hence it is necessary to clarify their phylogenetic relationships and infer possible reticulate relationships for the genus. In this study, genome-wide data of 43 species from Polygonatum and its closely related taxa were obtained by Hyb-Seq sequencing. The phylogenetic trees constructed from genome-wide nuclear and chloroplast sequences strongly supported the monophyly of Polygonatum with division into three major clades. A high level of incongruence was detected between nuclear and chloroplast trees as well as among gene trees within the genus, but all occurred within each major clade. However, introgression tests and reticulate evolution analyses revealed low level of gene flow and weak introgression events in the genus, suggesting hybridization and introgression were not dominant during the evolutionary diversification of Polygonatum in the Northern Hemisphere. This study provides important insights into reconstructing evolutionary relationships and speciation pattern of taxa from the north temperate flora.

SE-VisionTransformer: Hybrid Network for Diagnosing Sugarcane Leaf Diseases Based on Attention Mechanism.

Sugarcane is an important raw material for sugar and chemical production. However, in recent years, various sugarcane diseases have emerged, severely impacting the national economy. To address the issue of identifying diseases in sugarcane leaf sections, this paper proposes the SE-VIT hybrid network. Unlike traditional methods that directly use models for classification, this paper compares threshold, K-means, and support vector machine (SVM) algorithms for extracting leaf lesions from images. Due to SVM's ability to accurately segment these lesions, it is ultimately selected for the task. The paper introduces the SE attention module into ResNet-18 (CNN), enhancing the learning of inter-channel weights. After the pooling layer, multi-head self-attention (MHSA) is incorporated. Finally, with the inclusion of 2D relative positional encoding, the accuracy is improved by 5.1%, precision by 3.23%, and recall by 5.17%. The SE-VIT hybrid network model achieves an accuracy of 97.26% on the PlantVillage dataset. Additionally, when compared to four existing classical neural network models, SE-VIT demonstrates significantly higher accuracy and precision, reaching 89.57% accuracy. Therefore, the method proposed in this paper can provide technical support for intelligent management of sugarcane plantations and offer insights for addressing plant diseases with limited datasets.

Methylation Levels in the Promoter Region of FHIT and PIAS1 Genes Associated with Mastitis Resistance in Xinjiang Brown Cattle.

Mastitis causes serious economic losses in the dairy industry, but there are no effective treatments or preventive measures. In this study, the ZRANB3, PIAS1, ACTR3, LPCAT2, MGAT5, and SLC37A2 genes in Xinjiang brown cattle, which are associated with mastitis resistance, were identified using a GWAS. Pyrosequencing analysis showed that the promoter methylation levels of the FHIT and PIAS1 genes in the mastitis group were higher and lower, respectively, than those in the healthy group (65.97 ± 19.82% and 58.00 ± 23.52%). However, the methylation level of the PIAS1 gene promoter region in the mastitis group was lower than that in the healthy group (11.48 ± 4.12% and 12.17 ± 4.25%). Meanwhile, the methylation levels of CpG3, CpG5, CpG8, and CpG15 in the promoter region of the FHIT and PIAS1 genes in the mastitis group were significantly higher than those in the healthy group (p < 0.01), respectively. RT-qPCR showed that the expression levels of the FHIT and PIAS1 genes were significantly higher in the healthy group than those in the mastitis group (p < 0.01). Correlation analysis showed that the promoter methylation level of the FHIT gene was negatively correlated with its expression. Hence, increased methylation in the promoter of the FHIT gene reduces the mastitis resistance in Xinjiang brown cattle. Finally, this study provides a reference for the molecular-marker-assisted selection of mastitis resistance in dairy cattle.

Dental anxiety is related to postoperative symptoms in third molar surgery.

Purpose: To examine the association of preoperative dental anxiety with the severity of postoperative symptoms among patients undergoing lower third molar (LM3) extraction surgery. Materials and methods: We conducted a hospital-based prospective study with a sample size of 213 patients. All the patients underwent LM3 extraction surgery at the Stomatology Hospital of Tianjin Medical University. Preoperative dental anxiety was measured using the Dental Anxiety Scale for Third Molar Surgery (DAS-TMS) and classified into four categories: No anxiety, Some unease, Anxious, and Very anxious. The primary outcome was defined using the postoperative symptom severity scale on the seventh day after surgery. The patients' clinical characteristics, radiologic features, and surgery-related variables were used as control variables. Bivariate analysis involved Fisher's exact test and Kruskal-Wallis test. Multivariable logistic analysis was used to assess preoperative dental anxiety in relation to the severity of postoperative symptoms. We applied a two-piecewise regression model to examine the potential non-linear associations. Results: The mean (SD) dental anxiety score was 10.56 (3.84). The proportion of dental anxiety was as follows: No anxiety, 7.5%; Some unease, 46.9%; Anxious, 31.0%; Very anxious, 14.6%. The multivariable-adjusted ORs with 95% CIs of postoperative symptoms were 1.00 for No anxiety, 3.63 (0.90-14.68) for Some unease, 5.29 (1.25-22.33) for Anxious, and 4.75 (1.02-22.18) for Very anxious (P for trend = 0.047). The risk of serious postoperative symptoms increased with the dental anxiety level up to 7 points (adjusted OR 1.94, 95% CI 1.12-3.74; P = 0.012). When the dental anxiety level exceeded 7 points, the level of DAS-TMS was not associated with the risk of serious postoperative symptoms (OR 0.98, 95% CI 0.88-1.08; P = 0.756). Conclusions: Findings suggest that dental anxiety is associated with a risk of serious postoperative symptoms following LM3 removal. The degree of dental anxiety in patients before LM3 extraction surgery should be of concern to clinicians.

Factor Analysis of Genetic Parameters for Body Conformation Traits in Dual-Purpose Simmental Cattle.

In this study, we estimated the genetic parameters for 6 composite traits and 27 body conformation traits of 1016 dual-purpose Simmental cattle reared in northwestern China from 2010 to 2019 using a linear animal mixed model. To integrate these traits, a variety of methods were used as follows: (1) genetic parameters estimates for composite and individual body conformation traits based on the pedigree relationship matrix (A) and combined genomic-pedigree relationship matrix (H); (2) factor analysis to explore the relationships among body conformation traits; and (3) genetic parameters of factor scores estimated using A and H, and the correlations of EBVs of the factor scores and EBVs of the composite traits. Heritability estimates of the composite traits using A and H were low to medium (0.07-0.47). The 24 common latent factors explained 96.13% of the total variance. Among factors with eigenvalues ≥ 1, F1 was mainly related to body frame, muscularity, and rump; F2 was related to feet and legs; F3, F4, F5, and F6 were related to teat placement, teat size, udder size, and udder conformation; and F7 was related to body frame. Single-trait analysis of factor scores yielded heritability estimates that were low to moderate (0.008-0.43 based on A and 0.04-0.43 based on H). Spearman and Pearson correlations, derived from the best linear unbiased prediction analysis of composite traits and factor scores, showed a similar pattern. Thus, incorporating factor analysis into the morphological evaluation to simplify the assessment of body conformation traits may improve the genetics of dual-purpose Simmental cattle.

A Microfluidic System of Gene Transfer by Ultrasound.

Ultrasonic gene transfer has advantages beyond other cell transfer techniques because ultrasound does not directly act on cells, but rather pushes the gene fragments around the cells into cells through an acoustic hole effect. Most examples reported were carried out in macro volumes with conventional ultrasonic equipment. In the present study, a MEMS focused ultrasonic transducer based on piezoelectric thin film with flexible substrate was integrated with microchannels to form a microfluidic system of gene transfer. The core part of the system is a bowl-shaped curved piezoelectric film structure that functions to focus ultrasonic waves automatically. Therefore, the low input voltage and power can obtain the sound pressure exceeding the cavitation threshold in the local area of the microchannel in order to reduce the damage to cells. The feasibility of the system is demonstrated by finite element simulation and an integrated system of MEMS ultrasonic devices and microchannels are developed to successfully carry out the ultrasonic gene transfection experiments for HeLa cells. The results show that having more ultrasonic transducers leads a higher transfection rate. The system is of great significance to the development of single-cell biochip platforms for early cancer diagnosis and assessment of cancer treatment.

Attention mechanism-based deep learning method for hairline fracture detection in hand X-rays.

Wrist and finger fractures detection is always the weak point of associate study, because there are small targets in X-rays, such as hairline fractures. In this paper, a dataset, consisting of 4346 anteroposterior, lateral and oblique hand X-rays, is built from many orthopedic cases. Specifically, it contains a lot of hairline fractures. An automatic preprocessing based on generative adversative network (GAN) and a detection network, called WrisNet, are designed to improve the detection performance of wrist and finger fractures. In the preprocessing, an attention mechanism-based GAN is proposed for obtaining the approximation of manual windowing enhancement. A multiscale attention-module-based generator of the GAN is proposed to increase continuity between pixels. The discriminator and the generator can achieve 93% structural similarity (SSIM) as manual windowing enhancement without manual parameter adjustment. The designed WrisNet is composed of two components: a feature extraction module and a detection module. A group convolution and a lightweight but efficient triplet attention mechanism are elaborately embedded into the feature extraction module, resulting in richer representations of hairline fractures. To obtain more accurate locating information in this condition, the soft non-maximum suppression algorithm is employed as the post-processing method of the detection module. As shown in experimental results, the designed method can have obvious average precision (AP) improvement up to 7% or more than other mainstream frameworks. The automatic preprocessing and the detection net can greatly reduce the degree of artificial intervention, so it is easy to be implemented in real clinical environment.

Whole genome resequencing reveals the genetic contribution of Kazakh and Swiss Brown cattle to a population of Xinjiang Brown cattle.

Xinjiang Brown cattle is characterized by wide environmental adaptation from its female parent (Kazakh cattle) and good production performance from its male parent (Swiss Brown cattle). However, the genetic basis underlying these characteristics hasnotbeenexplored. Here we compared 50 genomes of Xinjiang Brown cattle to the genomes of other eight breeds worldwide to analyze patterns of genetic variation in the Xinjiang Brown cattle. We found canonical genomic characteristics of cross breed with the lowest linkage disequilibrium and the highest effective population size. At the global level, Xinjiang Brown cattle had 9.88% Kazakh cattle and 90.12% Swiss Brown cattle inheritance. Our local ancestry inference revealed the segments with the excess of Kazakh cattle blood enriched in genes or pathways involved in digestion,absorption,metabolism and disease. More importantly, we also observed the completely fixed haplotypes inherited from Swiss Brown cattle harboring genes (LCORL, GHR, MEF2D, PCSK1 and MSRB3), KEGG pathways, cattle QTLs or human NHGRIGWAS catalog related to body measurement and growth traits. Our findings will not only help understand the process of cross breeding but can provide basic materials for further QTL mapping and improvement of important traits in Xinjiang Brown cattle.

Plastid phylogenomic analyses of the Selaginella sanguinolenta group (Selaginellaceae) reveal conflict signatures resulting from sequence types, outlier genes, and pervasive RNA editing.

Different from the generally conserved plastomes (plastid genomes) of most land plants, the Selaginellaceae plastomes exhibit dynamic structure, high GC content and high substitution rates. Previous plastome analyses identified strong conflict on several clades in Selaginella, however the factors causing the conflictions and the impact on the phylogenetic inference have not been sufficiently investigated. Here, we dissect the distribution of phylogenetic signals and conflicts in Selaginella sanguinolenta group, the plastome of which is DR (direct repeats) structure and with genome-wide RNA editing. We analyzed the data sets including 22 plastomes representing all species of the S. sanguinolenta group, covering the entire geographical distribution from the Himalayas to Siberia and the Russian Far East regions. We recovered four different topologies by applying multispecies coalescent (ASTRAL) and concatenation methods (IQ-TREE and RAxML) on four data sets of PC (protein-coding genes), NC (non-coding sequences), PCN (the concatenated PC and NC), and RC (predicted RNA editing sites "C" were corrected by "T"), respectively. Six monophyletic clades, S. nummularifolia clade, S. rossii clade, S. sajanensis clade, S. sanguinolenta I clade, S. sanguinolenta II clade, and S. sanguinolenta III clade, were consistently resolved and supported by the characteristics of GC content, RNA editing frequency, and gene content. However, the relationships among these clades varied across the four topologies. To explore the underlying causes of the uncertainty, we compared the phylogenetic signals of the four topologies. We identified that the sequence types (coding versus non-coding), outlier genes (genes with extremely high |ΔGLS| values), and C-to-U RNA editing frequency in the protein-coding genes were responsible for the unstable phylogenomic relationship. We further revealed a significant positive correlation between the |ΔGLS| values and the variation coefficient of the RNA editing number. Our results demonstrated that the coalescent method performed better than the concatenation method in overcoming the problems caused by outlier genes and extreme RNA editing events. Our study particularly focused on the importance of exploring the plastid phylogenomic conflicts and suggested conducting concatenated analyses cautiously when adopting organelle genome data.

High-power, low-noise Brillouin laser on a silicon chip.

We realize a chip-based Brillouin microlaser with remarkable features of high power and low noise using a microtoroid resonator. Our Brillouin microlaser is able to output a power of up to 126 mW with a fundamental linewidth down to 245 mHz. Additionally, in the course of Brillouin lasing we observe an intriguing power saturation-like effect, which can be attributed to complex thermo-optic-effect-induced mode mismatch between the pump and Brillouin modes. To have a quantitative understanding of this phenomenon, we develop a model by simultaneously considering Brillouin lasing and the thermo-optic effect occurring in the microcavity. Of importance, our theoretical results match well with experimentally measured data.

Method development and validation for simultaneous determination of six tyrosine kinase inhibitors and two active metabolites in human plasma/serum using UPLC-MS/MS for therapeutic drug monitoring.

Over the past decades, therapeutic drug monitoring (TDM) of tyrosine kinase inhibitors (TKIs) and their main active metabolites has shown benefits in improving treatment efficacy and safety. Therefore, a sensitive, simple and economical ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the determination of six oral tyrosine kinase inhibitors (TKIs) and two active metabolites, including imatinib (IMA), N-desmethyl imatinib (NDIMA), sunitinib (SUNI), N-desethyl sunitinib (NDSUNI), regorafenib (REGO), nilotinib (NILO), dasatinib (DASA) and osimertinib (OSI) in human plasma/serum for therapeutic drug monitoring. The plasma/serum samples were deproteinated with acetonitrile after spiking with two deuterated internal standards (ISs, [2H8]-imatinib and [2H10]-sunitinib) and separated on a 40 °C ACQUITY UPLC® BEH C18 column (1.7 µm, 2.1 mm × 50 mm). The mobile phase was composed of acetonitrile (solution A) and water-formic acid-ammonium acetate (1 M) (994:1:5, v/v/v, solution B). Gradient elution was applied at a flow rate of 0.4 mL/min. Detection was carried out using a Triple Quad 5500 tandem mass spectrometer coupled with an electrospray ionization (ESI) source in positive mode. The method was validated over the calibration curve (CV) range of 2-400 ng/mL for NDSUNI and DASA, 2.5-500 ng/mL for SUNI, 10-2000 ng/mL NDIMA and OSI, 20-4000 ng/mL for NILO, 30-6000 ng/mL for REGO and 50-10000 ng/mL for IMA using linear regression and 1/x2 weighting. No difference was observed in the matrix effect (ME) among blank human plasma, hemolytic plasma, lipemic plasma, plasma spiked with ten commonly used drugs by cancer patients, or serum samples in general. The standard curve fitted by two standard curves of serum and plasma showed good linearity, and the precision and accuracy results of QC samples in both plasma and serum were acceptable and the concentration was comparable. To provide a clinical reference for the operation, the stability of the whole process from sample collection to drug detection was verified. SUNI and NDSUNI showed obvious photoisomerization under light exposure. Therefore, strict light protection was applied for all sample collection and handling steps of SUNI and NDSUNI. Compared with heparin anticoagulant tubes, the stability of the eight compounds in both whole blood and plasma was better in K3-EDTA and sodium citrate anticoagulant tubes. Given that all the analytes were stable in plasma at 4 °C for 48 h and in whole blood at room temperature for 48 h but OSI and REGO were unstable in whole blood and plasma at room temperature, samples should be centrifuged as soon as possible to be preserved as plasma at 4 °C when OSI or REGO is detected. In conclusion, this validated method can provide support for clinical practice, such as therapeutic drug monitoring (TDM) and pharmacokinetic studies of these six TKIs and two active metabolites.