RVT)” consisting of 25 postures using vibration stimulation of skeletal muscle and determined the feasibility of RVT for glycemic management. Thirty-one participants with obesity were enrolled in a controlled trial (CT) and experimental trial (ET) based on a 75 g oral glucose tolerance test (OGTT). During the CT, participants were required to rest in a quiet room. During the ET, the RVT program , consisting of 25 postures of relaxation and stretching on the vibratory platform, was performed for 40 min. Subsequently, the participants rested as in the CT. Subjective fatigue and muscle stiffness measurements and blood collection were conducted before and after RVT. In both the CT and ET, interstitial fluid (ISF) glucose concentrations were measured every 15 min for 2 h. The incremental area under the curve value of real-time ISF glucose during an OGTT was significantly lower in the ET than in the CT . Additionally, the levels of metabolic glucose regulators associated with myokines, muscle stiffness, and subjective fatigue significantly improved after RVT. This novel RVT suggests that it is effective in glycemic management with great potential to improve impaired glucose tolerance and T2D with obesity in the future.Because obesity is associated with impaired glucose tolerance and type 2 diabetes (T2D), it is important to manage the blood glucose level at an early stage. Nevertheless, people with obesity have significantly lower resistance to muscle fatigue after exercise and exercise adherence. Therefore, we developed a novel “Relaxing-Vibration Training ( The increasing prevalence of obesity directly and indirectly contributes to increased morbidity and mortality, including type 2 diabetes (T2D) . ConsideRecently, vibration training (VT), which contracts and relaxes skeletal muscles by vibration stimulation without a mass load or dynamic exercise, has received considerable attention. This mechanical stimulation uses proprioceptive spinal reflexes to induce an amyotrophic stretch reflex mediated by the muscle spindle and a type-Ia sensory fiber, thereby facilitating activity of homonymous α-motor neurons . It has RVT)” program that can be performed more comfortably and safely. As a first stage, we conducted an acute trial to verify the feasibility of a novel RVT for glycemic management. Using interstitial fluid (ISF) glucose concentrations during an oral glucose tolerance test (OGTT), blood markers, and muscle stiffness and fatigue were determined in middle-aged and older adults with obesity. We hypothesized that our practice of RVT has a more positive effect on subjective and biochemical indicators and has potential viability as a glycemic management program. The primary outcome was a change in the ISF glucose concentration, and the secondary outcomes were a change in blood markers, muscle stiffness, and fatigue.On the basis of these considerations, in this pilot study, we aimed to advance a previously general VT that requires high-intensity resistance posture and developed a novel “Relaxing Vibration Training . The study protocol was registered at the University Hospital Medical Information Network center (UMIN no. 000042787). We applied the devices, the Free-Style Libre Flash continuous glucose monitoring (FSL-CGM) system  and the Polar A370 fitness tracker , to study the participants’ bodies on the first day. Thereafter, the controlled trial (CT) was conducted on the fourth day and the experimental trial (ET) on the seventh day. Participants were required to fast for more than 10 h before each trial and were forbidden to consume alcohol or perform excessive exercise on the day before the trials. In both trials, participants ingested 225 mL soda-flavored solution  containing 75 g glucose and performed an OGTT lasting for 2 h, the ISF glucose concentration being recorded every 15 min. The details of the CTs and ETs are as follows  at the University of Tsukuba. The study objectives, design, criteria of inclusion and exclusion, assessments, practice of  follows : Participants were asked to fill out a questionnaire consisting of questions about age, sex, smoking, alcohol intake, and medical history, and to measure blood pressure and heart rate . Additionally, during the 2 h OGTT, participants were required to rest while measuring ISF glucose concentrations every 15 min on a chair in a quiet room. They were allowed to perform tasks such as reading a book, watching a movie, or operating a computer.RVT for 40 min, starting 15 min after the intake of a 75 g glucose solution. Subjective fatigue surveys, muscle stiffness measurements, and blood collection were conducted before and after RVT. Subsequently, the participants moved to a quiet room and rested as in the CT. In this study, we used a vibration machine  that can deliver three-dimensional harmonic vibration to the body. An expert with a power plate instruction certificate developed a novel RVT consisting of 25 postures of relaxation and stretching. The RVT was performed for 1 min per posture with a frequency of 50 Hz and an amplitude of 4 mm for 40 min on the vibratory platform, including preparation time for the next posture and rest  having one or more risk factors for metabolic syndrome, (4) active participation in the study. The participants were excluded if they (1) took neuropsychiatric drugs, (2) were prohibited from exercising by doctors due to serious diseases, including brain dysfunction, renal disease, liver dysfunction, heart disease, and peripheral angiopathy, (3) had an excessive alcohol intake (>60 g/day)  = 0.8) .The glucose concentration was determined using the FSL-CGM system, which has the advantage of being able to measure ISF glucose concentrations in real-time without blood collection through a sensor attached to the subcutaneous tissue. The FSL-CGM system consists of a glucose reader and sensor and the activation time for the sensor is 14 days, with the sensor’s high accuracy and convenience having been shown in previous studies ,14. Befo2) was calculated as body weight in kilograms divided by height in meters squared. Fat and muscle mass were determined using a bioelectrical impedance analyzer . We measured the waist circumference (WC), blood pressure, fasting blood glucose, high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) to confirm whether the participants displayed the corresponding risk factors for metabolic syndrome. Risk factors for metabolic syndrome were evaluated according to the Japanese standards as follows: abdominal obesity , high blood pressure , high fasting glucose (≥110 mg/dL or a history of diabetes), low HDL-C (<40 mg/dL), and hypertriglyceridemia (≥150 mg/dL or a history of hyperlipidemia) [BMI (kg/mpidemia) . The numBlood samples were collected from an antecubital vein and separated fractions were stored at −80 °C until further analysis. The free fatty acids (FFAs) and HDL-C levels were determined by enzyme method, lactate dehydrogenase (LDH), aspartate transaminase (AST), and creatine kinase (CK) levels by the Japan Society of Clinical Chemistry transferable method, fasting serum glucose level by the hexokinase-G-6-phosphate dehydrogenase method, high-sensitivity C-reactive protein (hs-CRP) level by fixed time assay method, and cortisol level by radioimmunoassay. We evaluated serum levels of fibroblast growth factor 21 , interleukin 6 , and myostatin  using commercial enzyme-linked immunosorbent assay kits.RVT in the ET using the Myoton® PRO , an accurate and reliable device for non-invasive digital palpation of superficial skeletal muscles. The location of the muscle to be measured was marked, and the probe of the Myoton® PRO device was vertically mounted on the surface of the measuring mark point as suggested by the manufacturer [RVT using the questionnaires “subjective symptoms of fatigue (Jikaku-sho shirabe)” and “body parts of fatigue (Hirou-bui shirabe)”, developed by the Japan Occupational Health and Occupational Fatigue Research Committee. The subjective symptoms of the fatigue questionnaire comprised 25 items divided into five categories . The body parts of the fatigue questionnaire investigated subjective fatigue levels for each body part, including the neck, shoulder, middle back, upper arm, forearm, lower back, hand, hip and thigh, lower leg and knee, and foot [Muscle stiffness of the trapezius, deltoid, biceps brachii, rectus femoris, biceps femoris, tibialis anterior, and medial gastrocnemius on the dominant side was determined before and after facturer . Three cand foot . RVT practice. The IAUC value is the sum of the areas of triangles and rectangles geometrically calculated using the elapsed time from baseline to 120 min and the response value of ISF glucose concentration. On the basis of the fasting blood sugar (0 min), the sum of increased values at 15 min (A), 30 min (B), 45 min (C), and 60 min  were multiplied by 15 and the sum of the increased values at 90 min (E) and 120 min  were multiplied by 30 {IAUC = (A + B + C + (D/2)) × 15((D/2) + E + (F/2)) × 30} [RVT, the effect size r was calculated as the Z statistic divided by the square root of the sample size (N) (Z/√N). All statistical analyses were performed using SPSS version 26.0 , with significance levels set to p < 0.05. The rate of change (Δ (%)) was calculated by subtracting the baseline value from the value after RVT practice and was expressed as a percentage of the baseline value. Linear mixed model analysis was applied to evaluate the differences in the intervention effect of the two trials on the change of ISF glucose concentrations. On the basis of significant interactions , this study performed post hoc analysis with Bonferroni correction. Wilcoxon signed-rank test was used only in the ET to compare changes in the incremental area under the curve (IAUC) of the ISF glucose concentration response, blood markers, muscle stiffness, and fatigue before and after )) × 30} . To veriAs shown in RVT practice. A significant interaction (p = 0.014) was observed between ISF glucose concentrations and the two trials, and according to the post hoc analysis, the ISF glucose concentrations in the ET was significantly reduced at 45 min (p = 0.019) and 60 min (p = 0.001). p = 0.047).RVT . Regarding the fatigue of body parts, the upper-body parts , lower-body parts , and total score of the whole body  decreased significantly. Additionally, drowsiness , instability , local pain or dullness , eyestrain , and total score of fatigue symptoms  decreased significantly after RVT  and immediately after the end of RVT (60 min). Overall, our study supports the positive result of previous studies that the glucose concentration can be controlled regardless of whether the period of skeletal muscle vibratory stimulation is acute or chronic. Vibration stimulation of skeletal muscles and tendons activates muscle spindles, which is termed “tonic vibration reaction”. It activates more motor neurons, which in turn activates motor units and the movement of actin and myosin, increasing muscle contraction . Such muRVT in the present study, there were great decreases in the ISF glucose concentration, FGF21, and myostatin. Contrary to the results of this study, it was reported that acute VT in both overweight and normal weight subjects displayed a time-dependent response in IL6, glucose, and insulin, but no change in myostatin [RVT may stimulate the hydrolysis of TG, which results in the release of FFAs to circulation and their oxidation in skeletal muscles. The elevated blood lipid level, observed in obesity and metabolic syndrome, is an adverse condition that leads to lipotoxicity and ectopic fat deposition in other organs, and consequently insulin resistance and impaired glucose metabolism [When muscle tissue is stimulated, it secretes myokines, which affect inflammation, glucose processing, and adipose tissue. FGF21 and myostatin, myokine factors, are known to be associated with obesity and insulin resistance . It was yostatin . By contyostatin . In the tabolism . Thus, eRVT, rather, the results that the muscle stiffness and fatigue improved showed satisfaction with this RVT. Moreover, because vibration stimulation of skeletal muscle has different physiological effects depending on the parameter settings , the protocol should be carefully set by a certified professional . In a prRVT in the present study significantly improved medial gastrocnemius stiffness, drowsiness, instability, local pain or dullness, eye strain, and whole-body fatigue. Moreover, after RVT practice in this study, the muscle-damage markers CK, AST, LDH, and hs-CRP did not change. In the participants with fibromyalgia, VT exercise  and conventional exercise significantly reduced pain and fatigue compared with the control group; however, stiffness and depression did not change. Additionally, it was insufficient to prove the effect of VT alone [RVT has advantages in the ability of glucose regulation and improvement of muscle fatigue while its practice is comfortable and safe.More importantly, we need to focus on the specific postures that were performed on the vibrating platform. High-intensity postures such as resistance exercise are difficult protocols for the elderly or diabetic patients in the high-frequency and high-amplitude settings. Because people with obesity and T2D have lower exercise adherence than healthy adults, strenuous resistance exercise accompanied by muscle fatigue is more likely to reduce exercise continuity ,4. The nVT alone . AnotherVT alone . DelayedVT alone . BecauseRVT, we were concerned about unexpected side effects in diabetic patients, including hypoglycemia and data instability. As mentioned in the introduction, we focused on people with obesity, because glucose regulation is important at an early stage from a prevention standpoint before the onset of IGT and T2D in obesity. Second, this pilot study was designed as an acute trial. The glucose concentration responds temporarily and sensitively to acute external stimuli such as diet and exercise. Therefore, we decided that it was necessary to first verify the novel RVT through an acute intervention trial. Because positive results were obtained in this situation, it is expected to show positive results in long-term interventions. Third, there was sampling bias and a relatively small sample size. Through the statistical software G* Power 3.1, 34 and 31 participants were estimated as a sample size by setting a power (1-β) of 0.8; however, it did not satisfy the sample size required for 0.95, which is the best power. However, a 1-β of statistical power indicates the possibility of Type II error (β) occurrence, with 0.8 meaning that even with 80% significant power, there is a 20% chance of not discriminating a significant difference. Additionally, it was reported that a power of ≥0.8 is more suitable for having a statistically significant difference [This pilot study has several limitations. First, it did not include diabetic patients exclusively. Because it is at an early stage to verify the feasibility of a novel fference . Fourth,RVT improved the ISF glucose concentration, FGF21 and myostatin levels, and muscle stiffness and fatigue in middle-aged and older adults with obesity. In the future, this novel RVT is expected to have a positive impact as a glycemic management program that can improve glucose regulation and fatigue in people with obesity, and further improve IGT and T2D. However, because this study was the result of an acute clinical trial, it is necessary to conduct additional studies on whether the same results can be obtained when long-term intervention with this novel RVT is conducted in diabetic patients in the future.In this pilot study, the acute practice of the novel


BackgroundCancer can significantly impact the nutritional status of patients, which can worsen related complications and reduce the effectiveness of treatment. Malnutrition is a common complication among cancer patients, especially among older adults. This study aims to determine the prevalence of malnutrition among hospitalized cancer patients and identify factors associated with malnutrition.Methodology and methodsThe cross-sectional descriptive study was performed on 296 cancer patients admitted to the National Oncology Referral Center in Sana'a, Yemen, between February 1 and March 31, 2022. Malnutrition was diagnosed through clinical assessment and screening tools such as screening tools, body mass index (BMI), weight loss percentage, and mid-upper arm circumference (MUAC). Descriptive statistics, chi-square, and multivariable logistic regression analysis were used to assess factors associated with malnutrition.ResultsOf the 296 patients included in the analysis, most were female 225 (76.0%), and the mean age was 45.8 years. The prevalence of undernutrition, as determined by the BMI score, was 48 (16.2%). The prevalence of malnutrition was higher in females 37 (16.4%) compared to males 11(15.5%). The data indicated that 49% of hospitalized patients had experienced weight loss of 5% or more. Multivariable logistic regression analysis showed that single  or widowed  marital status, weight loss , stomach cancer , and breast cancer  were associated with an increased risk of malnutrition.Conclusions The study highlights the importance of evaluating and addressing the nutritional status of cancer patients, especially at the beginning and during treatment, to improve their clinical outcomes. Prospective studies are recommended to further investigate this issue. Cancer is a major cause of death worldwide, with more recent estimates of 10 million deaths in 2020. Developing countries face particularly unfavorable outcomes due to limited access to healthcare and increased risk factors such as political and economic instability, as well as lifestyles transitioning towards Western patterns .The Eastern Mediterranean region (EMR) comprises 22 countries, with a population of almost 600 million people, spanning from Morocco in the west to Pakistan in the east. The disease burden in the EMR has changed in the past four decades from primarily communicable diseases to non-communicable diseases, including cancer and cardiovascular diseases , similarDue to limited diagnostic and clinical resources, poor quality of medical records, and ongoing civil war for nine years, the accurate incidence of cancer in Yemen is unknown, leading to the ambiguity of the burden of cancer at the national level. Therefore, a national malnutrition for cancer patient registry is required in the Republic of Yemen. The purpose of this study is to determine the prevalence of malnutrition among hospitalized cancer patients and identify factors associated with malnutrition.Study design and setting This study is an observational, descriptive cross-sectional study, which was conducted in Sana'a, the capital city of Yemen, between February 1 and March 31, 2022. Sana'a City, the capital city of Yemen is situated in the center of the country at an altitude of 2,300 m. The city has a population of 3,181,655 people , and it The study was performed between February 1 and May 31, 2022, at a National Oncology Referral Center, for oncology in northeastern Yemen, located in Sana’a City of Yemen. Eligible participants included all members of the general population aged 18 and older who were willing to participate in the study. Those patients hospitalized for more than 24 hours, who were fed exclusively through catheter or ostomies for more than 24 hours, with consumptive disorders, or pregnant were excluded from the sample.The study started after its approval by the Medical Research Committee (MRC) Ethics of the cited Al Razi University number 79, on 15.01.2022.Sample size and sampling methodThe study employed a convenience sampling approach, recruiting participants from the available pool at the time of the study. Specifically, the study participants were recruited from an outpatient clinic located at the National Oncology Center in Sana'a City, Yemen.The sample size was calculated using the formula2 *p*(1-p))/e2 (1)n= , p = prevalence (80%) , considering the cutoff point of % WL ≥5 in the past six months. The body mass index (BMI) was obtained by the equation  and was classified according to the recommendations of the World Health Organization (WHO) for adults and the Pan American Health Organization (PAHO)/WHO for the elderly [/ x 10) (2)Statistical analysisTo analyze the data, cancers were classified into six groups based on the location of the tumor, including upper gastrointestinal tract, lower gastrointestinal tract, reproductive tract, lung, breast, and other cancers that did not fit into any of the former classifications. Therapeutic variables were stratified into clinical treatment, surgical treatment, hormonal treatment, and palliative treatment. Categorical variables were presented using absolute and relative frequencies, and continuous variables were summarized using mean and standard deviation measures.Pearson's chi-square test was used for bivariate analysis, and a two-sided test with a significance level of alpha <0.05 was employed. Multivariable logistic regression was used to estimate adjusted odds ratios.Initially, all variables were included in the analyses, but only variables associated with malnutrition with a p-value <0.25 were retained in the model. The step-wise backward elimination method was applied to develop the final multivariable logistic regression model . A two-sPatients’ sociodemographic characteristicsDuring the study period, a total of 296 cancer patients were admitted, with the majority of the sample being female 225 (76.0%). The mean age of the patients was 45.8±14.5 years, with 151 (51%) falling within the age range of 41-60 years. Most of the participants were married 220 (74.3%), and 137 (46.3%) had no formal education. Among the educated participants, 62 (20.9%) had a primary/intermediate level of education, and six (2%) had a diploma. The highest percentages of respondents were from Dhamar , Taiz , Ibb , and the Capital City Sana'a , while the lowest frequencies were from Al-Jawf and Aden . Moreover, 125 patients (42.2%) were from urban areas, 91 (30.7%) from rural areas, and 80 (27%) from suburban areas.The family income of the participants varied from YR 3,500 to YR 1,200,000, with the majority of them belonging to the income group of <YR 50,000 (36.5%). The mean SD monthly household income was YR 82,204.4±100,439.5. Regarding family history of cancer, 82 patients (28%) reported having a family history of cancer, with the father/mother being the most common relative with a history of cancer 29 patients (35%), followed by uncles 27 patients (33%).Regarding smoking status, 104 patients (35.1%) were smokers/ex-smokers, 172 patients (58.1%) reported no qat chewing, and 123 (41.6%) reported having chewed qat  and reproductive system cancer (13.2%). Among respondents aged 60 years or older, breast cancer 17 (26.6%) and reproductive system cancer 12 (18.8%) were the most common types. In terms of gender, tumors of the upper gastrointestinal tract were more prevalent among males 16 (22.5%), while breast tumors were more prevalent among females 122 (54.2%).The sample showed a considerable prevalence of weight loss ≥5% in breast tumors 52 (35.9%), followed by tumors of the upper gastrointestinal tract 22 (15.2%). Low weight was also prevalent among respondents with breast tumors 12 (25.0%) and tumors of the upper gastrointestinal tract 10 (20.8%)  of cancer patients. These variables were entered into a multivariable binary logistic regression model to control for the effect of confounding factors.In the multivariable logistic regression analysis, age (year) was found to be associated with nutritional status, with younger patients having a 96% decreased chance of malnutrition . Patients who were single or widowed were 12.93 and 11.51 times more likely to be malnourished than the reference group of cancer patients , and , respectively.Patients who experienced weight loss with a percentage of ≥5% were 7.56 times more likely to be malnourished . Finally, patients with stomach and breast cancer were 6.77 times more likely to be malnourished than those with other types of cancer , and patients with breast cancer were 2.60 times more likely to be malnourished than those with other types of cancer  . The finding of this study was lower than the study done in Vietnam (84.7%) , Brazil .2%. The The difference in prevalence between the current study and previous studies could be attributed to variations in socio-demographic characteristics, study population, economic status, impaired immune function, and expanded health service provision, as well as the use of different assessment tools. The current prevalence observed in the current study may be due to the side effects associated with chemotherapy, including oral mucositis, intraoral mucositis, intraoral infection, dry mouth, salivary gland inflammation, mucosal bleeding, and intraoral hemorrhage, which can hinder nutritional intake and weaken the immune response. Furthermore, gastrointestinal cancer represents a percentage of 11.5%, which may have hampered their ability to eat.Additionally, most of the clients in the current study were suffering from gastrointestinal cancer, which could potentially affect their ability to consume food.Analysis of the percentage of weight loss revealed that 49.0% of the participants experienced a weight loss of more than 5% over the course of six months, which is considered an early sign of malnutrition. In comparison to a study conducted in France, a lower proportion of weight loss was reported, with 17.4%, 12.6%, and 10.9% of the patients experiencing weight loss of 5-10%, 10-15%, and greater than 15%, respectively . A similThe presence of these conditions, such as weight loss, is indicative of inadequate nutritional intake due to the interference of the tumor itself on the body's functioning and the side effects of treatment interventions, especially chemotherapy. Additionally, the current findings revealed that 52.0% and 43.6% of the participants experienced nausea and loss of appetite, respectively.In this study, more than 83.3% of the patients with cancer had comorbidities, such as arthritis, hypertension, diabetes, cardiovascular diseases, chronic kidney disease, and depression. The literature supports the finding that co-prevalence of these conditions is common in cancer patients and can be explained by shared risk factors, including being overweight, having a sedentary lifestyle, smoking, alcoholism, and poor eating habits. It is important to note, however, that chemotherapy can cause changes in an individual's blood pressure and can have a significant impact on the occurrence of hypertension in this population ,20.In this study shows that previous or current radiotherapy and chemotherapy were independently associated with an increased risk of malnutrition. This may be due to reduced food intake and/or increased metabolism. Previous studies declared that the association between radiotherapy/chemotherapy and an increased risk of malnutrition may have practical consequences.Better knowledge of these side effects of chemotherapy-induced and their related chemosensory implications is warranted to generate evidence-based clinical, preventive, and therapeutic guidelines.In the bivariate analysis age by years, sex, complicated with chronic disease, hypertension, cardiovascular disease, arthritis, smoking, stomach cancer, breast cancer, and lung cancer were associated with nutritional status (underweight) of cancer patients.The variables mentioned above were entered into a multivariable binary logistic regression model to control the effect of confounders.Multivariable logistic regression in our study showed that being young reduces the chance of malnutrition by almost 96%. This association was expected, as it is well-known that advanced age predisposes individuals to nutritional deficiencies, particularly in hospitalized patients . In the Our study found that stomach cancer was associated with malnutrition among cancer patients, which is consistent with studies conducted in Latin America  and VietFurthermore, our study identified that women with breast cancer were significantly associated with malnutrition. This finding is consistent with a previous study .Cancer-induced malnutrition is caused by multiple factors that differ significantly from those associated with simple starvation. These factors include mental health issues, poor food intake, gastrointestinal tract dysfunction, increased energy needs, decreased physical activity, and changes in metabolism in various organs or tissues . Our stuThe prevalence of malnutrition among adult cancer patients was 16.2%. Malnutrition was higher among female cancer patients than among males. Single or widowed marital status, weight loss, stomach cancer, and breast cancer were found to be significant factors for malnutrition. Therefore, it is crucial to quickly identify individuals with malnutrition or at risk of malnutrition by increasing healthcare professionals' awareness and implementing suitable nutritional screening programs at the beginning and throughout the treatment process. Further qualitative and longitudinal studies are needed to explore the state of malnutrition in cancer patients.


Henan is the province with the greatest wheat production in China. Although more than 100 cultivars are used for production, many cultivars are still insufficient in quality, disease resistance, adaptability and yield potential. To overcome these limitations, it is necessary to constantly breed new cultivars to maintain the continuous and stable growth of wheat yield and quality. To improve breeding efficiency, it is important to evaluate the genetic diversity and population genetic structure of its cultivars. However, there are no such reports from Henan Province. Therefore, in this study, single nucleotide polymorphism (SNP) markers were used to study the population genetic structure and genetic diversity of 243 wheat cultivars included in a comparative test of wheat varieties in Henan Province, aiming to provide a reference for the utilization of backbone parents and the selection of hybrid combinations in the genetic improvement of wheat cultivars.In this study, 243 wheat cultivars from Henan Province of China were genotyped by the Affymetrix Axiom Wheat660K SNP chip, and 21 characteristics were investigated. The cultivars were divided into ten subgroups; each subgroup had distinct characteristics and unique utilization value. Furthermore, based on principal component analysis, Zhoumai cultivars were the main hybrid parents, followed by Aikang 58, high-quality cultivars, and Shandong cultivars. Genetic diversity analysis showed that 61.3% of SNPs had a high degree of genetic differentiation, whereas 33.4% showed a moderate degree. The nucleotide diversity of subgenome B was relatively high, with an average π value of 3.91E-5; the nucleotide diversity of subgenome D was the lowest, with an average π value of 2.44E-5.The parents used in wheat cross-breeding in Henan Province are similar, with a relatively homogeneous genetic background and low genetic diversity. These results will not only contribute to the objective evaluation and utilization of the tested cultivars but also provide insights into the current conditions and existing challenges of wheat cultivar breeding in Henan Province, thereby facilitating the scientific formulation of breeding objectives and strategies to improve breeding efficiency.The online version contains supplementary material available at 10.1186/s12870-023-04537-9. Although there are more than 100 cultivars planted for crop production, many remain insufficient in quality, disease resistance, adaptability, and yield potential. Moreover, due to the interference of natural or human factors such as selection, mutation, migration and genetic drift, the characteristics of cultivars may change, and the appropriate planting time of a cultivar during production is limited. Therefore, it is necessary to continuously breed new cultivars to overcome these limitations, cope with and meet changing natural and production conditions, and maintain a continuous, stable increase in wheat yield. However, to improve breeding efficiency and accelerate the breeding process, it is necessary to characterize population genetic structure and genetic diversity [Henan is the province with the greatest wheat production in China, with an annual cultivated area of more than 5.69 million ha, accounting for approximately 24.8% of the national planting area and 28.3% of the national yield  [Aegilops tauschii [Agropyron cristatum L.) [Triticum dicoccoides) [Triticum durum) [Triticum aestivum) [Previous studies have examined the genetic diversity and population structure of Urartu wheat ( urartu) , Aegiloptauschii , crestedatum L.) , wild emccoides) , durum wm durum) –31, syntm durum) , and comestivum) –41 usingestivum) , 43, aimFigure Additionally, π is a measure of nucleotide diversity in a population. A larger π indicates higher nucleotide diversity, whereas a lower π specifies lower nucleotide diversity between the two loci. Figure The tested cultivars in this study were bred by cross-breeding of cultivars widely planted in the past and now with varieties bred abroad or in other parts of China or new materials generated by breeding units themselves. Therefore, there is a certain association between these cultivars. If the genotype information for these cultivars can be transformed into independent gene type information, insights may be provided regarding the main parent resources for breeding, thereby allowing for cross combination selection to be purposefully carried out to facilitate increased breeding efficiency. To this end, PCA of 243 tested cultivars was performed using GAPIT software . Figure PCA of the 243 cultivars showed that eight principal components (eigenvectors) had relative contribution rates > 2%. The relative contribution rates of principal components 1–8 were 10.15%, 8.59%, 6.41%, 3.77%, 2.99%, 2.90%, 2.55%, and 2.28%, respectively, and the cumulative contribution rate was 39.64%. Table In this study, all tested cultivars were bred by crossing. According to analysis of the hybrid combinations of the cultivars corresponding to the ten elements with the largest absolute coefficient values for the top 8 principal components Table , the mosWe then determined the genetic structure of the population using 395,783 high-quality SNP markers. Using ADMIXTURE software , the minAnalysis of variance Table  showed t2 and an average 1,000-kernel weight of 44.25 g. Additionally, the highest degree of lodging and lodging area were recorded in this subgroup. The severity of powdery mildew was 3.44, which was the highest among all subgroups. The yield was 7.86 t/ha , average lodging degree, average lodging area, degree of freezing injury, and average yield (7.60 t/ha). The average kernel number per spike was 39.51 grains  was the second highest in each subgroup. The cultivars had the lowest average lodging degree and lodging area. Fusarium head blight resistance was the worst (3.75 grade) among all subgroups . Genetic diversity analysis showed that 61.3% of the SNPs had high genetic differentiation (Fst > 0.2); 69.7% had high genetic differentiation in subgenome B, whereas 52.0% and 56.6% had high genetic differentiation in subgenomes A and D, respectively. The analysis of nucleotide diversity showed that the nucleotide diversity of subgenome B was relatively high, with an average π value of 3.91E-5; the nucleotide diversity of subgenome D was the lowest, with an average π value of 2.44E-5. Compared with that reported by Hao et al. (14), the π value was slightly smaller, which may be related to the use of different materials and SNP typing chips. These results indicate that after years of repeated selection, there is a higher degree of genetic differentiation with low nucleotide diversity in newly bred cultivars in Henan Province. Hence, it is difficult to breed breakthrough cultivars by crossing these cultivars with each other. However, the “best” cultivars can be used as parents for hybridization with foreign varieties or to introduce superior genes from related plants to breed new cultivars with breakthrough yield potential, disease resistance, stress resistance, and quality.PCA showed that Zhoumai cultivars were the most commonly used parents for breeding the modern wheat cultivars present in Henan Province. Zhoumai cultivars are new wheat cultivars bred by the Zhoukou Academy of Agricultural Sciences since the 1980s and are widely planted in the Huang Huai wheat-producing region. Zhoumai cultivars are high yielding and disease resistant. According to statistical data published in 2016 , five of the six most cultivated wheat cultivars (> 333 kha) in Henan Province possess genetic materials from Zhoumai cultivars. Among 11 wheat cultivars with an annual planting area of 67 ~ 333 kha, six possess genetic materials from Zhoumai. Since 2011, 64% of the semiwinter wheat cultivars certified by Henan Province have been found to possess genetic materials from Zhoumai, whereas 67% of the semiwinter wheat cultivars certified by the state have genetic materials from Zhoumai. Thus, it is evident that Zhoumai cultivars are the most important backbone parents in wheat cross-breeding in Henan Province.The second most commonly used parent for breeding modern cultivars was Aikang 58, which is a high-yield dwarf wheat variety with multiple resistance and wide adaptability. The largest annual planting area is more than 3 million ha, and the cumulative planting area is more than 20 million ha. Many breeding units have bred 128 approved wheat cultivars with Aikang 58 as a parent.The third type is high-quality parents, which differ from those prevalent more than 20 years ago, although most breeding work requires further improvement of yields and stress resistance. However, increasing attention has been given to improving quality, which has become one of the main wheat breeding objectives in Henan Province. Nevertheless, quality should not compromise yield, and vice versa. Indeed, cultivars with favorable quality but poor yield will not be accepted by farmers owing to profitability issues. Therefore, breeders must attempt to breed cultivars with an acceptable combination of quality, yield, and stress resistance.The fourth most common parent cultivars were those from Shandong, which is adjacent to Henan Province and has similar ecological and climatic conditions. Thus, Shandong cultivars have good adaptability in Henan Province, as well as good quality and great yield potential. Hence, Shandong wheat cultivars have made significant contributions to the genetic diversity of modern Henan wheat cultivars.Based on analysis of the cross combinations of the cultivars corresponding to the ten elements with the largest absolute coefficient values for each PC in PCA, it can be seen that the parents used in wheat cross-breeding in Henan Province are quite similar, with a relatively homogenous genetic background. Thus, considering the low genetic diversity of modern wheat cultivars in Henan Province, new genetic materials from other countries or regions should be used to improve diversity and production performance, while genomic techniques, such as hybridization, genetic modification, and cloning, could be employed to enhance the genetic diversity and performance of wheat cultivars.The objective evaluation of cultivars and classification of subgroups are important for the rational distribution and application of cultivars. Previously, wheat populations were clustered based on the phenotypic traits of cultivars. However, most phenotypic traits of wheat are quantitative traits, which are easily affected by environmental conditions. The expression of these phenotypic traits varies with location and time. For the same population, even if the same group of phenotypic traits and classification methods are used, the clustering results will differ if the test time or location differs. However, PCA clustering based on genotype diversity is more stable and reliable, as the results are not affected by the test time or location. With the advancement of molecular biology techniques, researchers have identified loci of interest using genetic markers. In the present study, the Affymetrix Axiom Wheat660K SNP chip was employed to examine the genetic diversity and population structure of 243 modern wheat cultivars in Henan Province. After performing a series of quality control steps, 395,783 SNP markers were retained, and 243 cultivars were assigned to ten subgroups. Except for susceptibility to powdery mildew and sharp eyespot, the remaining phenotypes significantly differed among subgroups. However, members of the subgroups shared certain characteristics and traits.Subgroup III had the highest yield and disease resistance. The cultivars in this subgroup, as well as those in other subgroups with various beneficial characteristics, including high yield, high quality, stress resistance, and disease resistance, can be popularized in production following regional testing, production testing, and approval. Of course, these cultivars can also be used as parents, and superior cultivars can be bred through further genetic improvement. For instance, to increase the number of spikes per unit area, it is important to focus on using cultivars in subgroups I and II as parents. Moreover, to increase grain weight, cultivars in subgroups IV, VI, and IX would be beneficial as parents. To reduce plant height, cultivars in subgroups II and VIII should be considered parents, and to improve cold tolerance, those in subgroups II and V can be used. Furthermore, to improve lodging resistance, cultivars in subgroups VI, VII, and IX would represent effective parents. Finally, if large panicles and numerous grains are desired, the cultivars in subgroup VII should be used as parents. Hence, collectively, the findings of this study provide comprehensive insights for the selection of optimal cultivars with excellent comprehensive traits or specific breeding objectives in Henan Province and other regions.In summary, the genetic diversity and population structure of 243 modern wheat cultivars in Henan Province were analyzed by SNP genotyping and phenotypic data of related traits in this study. A higher degree of genetic differentiation was observed among different cultivars. That is, SNPs with a high degree of genetic differentiation (Fst > 0.2) accounted for 61.3%, whereas those with a moderate degree (Fst = 0.05 ~ 0.2) accounted for 33.4% of all SNPs. Based on the analysis of cross combinations of cultivars corresponding to the ten elements with the largest absolute coefficients for each PC in PCA, Zhoumai cultivars were identified as the main parent of the tested cultivars, followed by Aikang 58, high-quality cultivars, and Shandong cultivars. Through population genetic structure analysis, these cultivars were further divided into ten subgroups, each with its own distinct characteristics and unique utilization value. Hence, these results not only contribute to the objective evaluation and utilization of the tested varieties but also further our current understanding regarding the current challenges affecting wheat cultivars in Henan Province. This inspires us to scientifically formulate breeding objectives and strategies to improve the efficiency and speed of the breeding process. The primary limitation of this paper is the near exclusive inclusion of semiwinter cultivars. Thus, future cultivar investigations should be extended to those prevalent in areas with weak spring and spring production and larger areas.Triticum aestivum L.) cultivars included in the Comparative Test of Wheat Varieties in Henan Province, and Zhoumai 18 was used as the control cultivar. The tested cultivars were randomly arranged. To reduce experimental error, the experiment was based on the interval contrast design, a control was set every six cultivars, 210 basic seedlings/m2 were planted, and the area of each plot was at least 13 m2. Sowing occurred on approximately October 10, and harvest occurred in early June of the following year. Field management procedures, including fertilization, soil preparation, sowing, and irrigation, were carried out according to general field experimentation protocols.Field experiments were conducted at Xihua , Luohe , Xuchang , Xinxiang , Wenxian , Huixian , and Nanle  in Henan Province in 2015–2016. The experimental materials were 243 bread wheat  were selected within the plot, and the effective spike number was determined and converted to spikes/mKernel number per spike: Before harvest, 50 single spikes were randomly harvested from each plot, mixed and threshed, the total kernel number was determined, and the average grains per spike was calculated.1000-Kernel weight: One thousand seeds were randomly weighed (unit: g) twice, and the average value was taken , accurate to one decimal place.Fertility period: The fertility period was measured as the number of days from emergence to maturity.Seedling habit: Seedling habit was observed during the tillering period, with three levels: creeping, semicreeping and upright; expressed as 1, 2, and 3, respectively.Maturity performance: This trait was ranked as good, medium, and poor based on stem and leaf yellowing; expressed as 1, 3, and 5, respectively.Plant height: Plant height was measured from the ground to the top of the spike, excluding awns, in centimeters.Lodging resistance: This trait was measured twice as initial lodging and final lodging, recording the lodging date, degree and area, summarized by the final lodging data. The lodging area was the percentage of lodged area relative to the plot area. The lodging degree was recorded at five levels: no lodging; slight lodging, plant tilting angle less than or equal to 30°; moderate lodging, tilting angle 30° to 45° (inclusive of 45°); heavy lodging, tilting angle 45° to 60° (inclusive of 60°); and severe lodging, tilting angle above 60°; expressed as 1, 2, 3, 4, and 5, respectively.Cold resistance: According to the above ground freeze damage recorded in both the overwintering and spring stages, the plants were divided into five levels: no freeze damage, leaf tips turned yellow from freezing, half of the leaf blade frozen until dead, entire leaf blade withered, and plant or most tillers frozen until dead; expressed as 1, 2, 3, 4, and 5, respectively.Powdery mildew: This trait was recorded as five levels during the outbreak period during heading: no visible symptoms on leaves, disease on basal leaves, lesions spread to middle leaves, lesions spread to flag leaves, and lesions spread to spikes and awns; expressed as 1, 2, 3, 4, and 5, respectively.Sharp eyespot: Stem peeling was observed during the peak period of onset after full heading, with this trait recorded as five levels: no symptoms, leaf sheath infected but not penetrating the stem, lesions penetrating the stem but not to more than 1/4 (including 1/4) of the stem circumference, lesions penetrating the stem to between 1/4 and 3/4 (including 3/4) of the stem circumference, and lesions penetrating the stem to more than 3/4 of the circumference; expressed as 1, 2, 3, 4, and 5, respectively.Fusarium head blight: The severity of infection on the spikelets was visually estimated, and this trait was recorded as five levels: no diseased spikes, 1/4 (including 1/4) or fewer spikelets diseased, 1/4 to 1/2 (including 1/2) of spikelets diseased, 1/2 to 3/4 (including 3/4) of spikelets diseased, and more than 3/4 of spikelets diseased; expressed as 1, 2, 3, 4, and 5, respectively.Rust diseases: Leaf rust and stripe rust were recorded in terms of prevalence, severity, and reaction type. Incidence was estimated as the percentage of diseased leaves relative to the total number of leaves. Severity was estimated as the percentage of the lesion distribution relative to the leaf  area. Reaction type was divided into five levels: immune, completely symptomless, or with very small pale spots; highly resistant, leaves with yellow‒white dead spots or very small spore masses, surrounded by obvious dead spots; moderately resistant, summer spore masses few and scattered, surrounded by fading green or dead spots; moderately susceptible, more summer spore masses, surrounded by fading green; and highly susceptible, many large summer spore masses, with no fading green surrounding; expressed as 1, 2, 3, 4, and 5, respectively.According to the standard NY/T 1301–2007 guidelines  issued by the Ministry of Agriculture, China, we examined the expression of 21 traits, including seedling habit and plant height, in each cultivar at all 7 test sites. The observation and recording standards for each trait are as follows:A joint analysis of variance was conducted on the results from each test location by phenotypic trait based on a fixed model. The total variation was partitioned among five factors: test locations, subgroups, test location × subgroup, cultivars within subgroup, and experimental error. Multiple comparisons between different subgroups were conducted using Duncan's new multiple-range test. Statistical calculations were implemented in SPSS 26.0.http://wheat.pw.usda.gov/GG2/index.shtml) from the Affymetrix Axiom Wheat660K SNP chip at Beijing CapitalBio Technology Co., Ltd. This chip was developed by Jizeng Jia et al. at the Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, and Affymetrix by sequencing 192 wheat varieties, including 60 modern cultivars, 72 local varieties, 30 wild one-grain varieties (Triticum boeoticum), and 30 varieties of Aegilops tauschii. Approximately 90% of the 630,517 SNP markers were polymorphic [In this study, an improved sodium dodecyl sarcosine method , 46 was ymorphic , 43. TheSNP markers with a heterozygosity rate > 10%, sample detection rate < 90%, and minimum allele frequency < 0.02 were excluded using PLINK V1.07 software . FinallyRaw data for the 395,783 SNPs were converted into VCF format. The population differentiation index (Fst) and nucleotide diversity index (π) were calculated using VCFtools V4.1 software . The calPCA is a statistical method that transforms multiple interrelated characteristic indexes into a few independent comprehensive characteristic indexes. It not only uses the eigenvalue to reflect the relative contribution rate of each principal component but also uses the coefficients of each element of the principal component (eigenvector) to analyze the main features of each principal component. It is also an effective method for visualizing the distribution of different genotypes. PCA of the 243 wheat cultivars in Henan Province was performed using the GAPIT 3.0 software package developed by Cornell University in R studio , 48, 49.K values ranging from 1 to 20 were tested until the minimum validation error was achieved. In total, 20 independent runs were performed for each K value.We compared the genetic structure of the samples using ADMIXTURE V1.3 software  based onAdditional file 1: Table S1. Cross combinations of the cultivars corresponding to the ten elements with the largest absolute values of the coefficients of their elements corresponding to top 8 principal components.Additional file 2: Table S2. Tested cultivars from each subgroup.Additional file 3: Table S3. The probability that the tested cultivars belong to a subgroup.


APC, SYNE1, TP53, and TTN genes. Among the differently methylated and expressed genes were those involved in cell adhesion, extracellular matrix organization and degradation, neuroactive ligand-receptor interaction. The top up-regulated microRNAs were hsa-miR-135b-3p and -5p, and the hsa-miR-200-family while the hsa-miR-548-family belonged to the top down-regulated. MmCRC patients evinced higher tumor mutational burden, a wider median of duplications and deletions, and a heterogeneous mutational signature than SmCRC. Regarding chronicity, a significant down-regulation of SMOC2 and PPP1R9A genes in SmCRC compared to MmCRC was observed. Two miRNAs were deregulated between SmCRC and MmCRC, hsa-miR-625-3p and has-miR-1269-3p. The combined data identified the IPO5 gene. Regardless of miRNA expression levels, the combined analysis resulted in 107 deregulated genes related to relaxin, estrogen, PI3K-Akt, WNT signaling pathways, and intracellular second messenger signaling. The intersection between our and validation sets confirmed the validity of our results. We have identified genes and pathways that may be considered as actionable targets in CRCLMs. Our data also provide a valuable resource for understanding molecular distinctions between SmCRC and MmCRC. They have the potential to enhance the diagnosis, prognostication, and management of CRCLMs by a molecularly targeted approach.Despite distant metastases being the critical factor affecting patients’ survival, they remain poorly understood. Our study thus aimed to molecularly characterize colorectal cancer liver metastases (CRCLMs) and explore whether molecular profiles differ between Synchronous (SmCRC) and Metachronous (MmCRC) colorectal cancer. This characterization was performed by whole exome sequencing, whole transcriptome, whole methylome, and miRNAome. The most frequent somatic mutations were in  Metastasis, the process of spreading cancer cells from the primary site to distant organs, is a major cause of cancer mortality . The livWhen the CRCLMs are not treated, the disease exhibits an unfavorable prognosis with the median overall survival (mOS) of less than 12 months. Even with the use of aggressive treatment , the overall survival (OS) does not exceed 13-18 months . The onlPatients with CRCLMs form a heterogeneous group. We can distinguish between Synchronous (SmCRC) and Metachronous (MmCRC) colorectal cancers. There is, however, still no consensus on the definition of SmCRC and MmCRC as used in the context of CRCLMs. Although by definition, all metastases are synchronous (occult or detectable at diagnosis), most definitions include detection at or before diagnosis or surgery of the primary tumor , while sOver the past few decades, numerous preclinical and clinical studies have been conducted to uncover the underlying mechanisms of CRCLMs formation. However, only a limited number of results from these studies reached clinical practice, such as assessing PD-L1 expression in tumor cells using immunohistochemistry to provide prognostic information and predict response to treatment with PD-1/PD-L1 inhibitors . The lacIn the present study, the high-throughput approaches with comprehensive bioinformatic analysis provided us with a platform for the analysis of metastatic tissue, both SmCRC and MmCRC, to find relevant hub genes and pathways and understand the molecular mechanism and characteristics of CRCLMs. In addition, we also investigated clinicopathologic information of SmCRC and MmCRC patients to improve patients´ management and follow-up.In this prospective single-center study, histologically confirmed CRCLM patients were recruited at the Department of Oncology and Radiotherapy  between June 2019 and December 2021. All CRCLM cases were monitored regularly until May 31st, 2022. Patients with any personal history of previous other malignancy or with CRC-associated well-defined inherited syndromes  were excluded from the study. In total, 10 CRCLM patients were included in the study.Biological samples were immediately put in an RNA Later stabilization reagent  and stored at − 80°C. Clinical data were collected from all subjects recruited in the study. The clinicopathological data for the patients recruited are reported in The local ethics committees at the Faculty Hospital in Hradec Kralove, Czech Republic  and the Institute of Experimental Medicine, Prague, Czech Republic  approved the study. All patients provided written informed consent.The colon mucosa from healthy individuals the fresh frozen samples were collected consecutively during the planned colonoscopy and were described in Jungwirth et al. .The first cells invading the liver came from the primary CRC tumor. They thus also carry the genome/transcriptome of the primary tumor, respectively colorectum. Thus, comparing the genetic/molecular background of CRC cells with non-tumor liver cells could lead to results focused on organ specificity rather than on the distinctiveness of the metastasis relative to the healthy tissue. For this reason, we included a set of control colon tissues into the analyses apart from the patients’ paired samples (metastases and adjacent liver tissue). Overall, control colorectal tissues (n=10) were used to eliminate false positive results from contamination of liver tissue. These tissues were used from healthy individuals with no previous or current cancer. This correction was used for DNA methylation assessment and RNA seq analysis. A more detailed description of this correction is explained in the bioinformatics section of the methods.DNA isolation, quality and quantity analyses were all performed according to the manufacturer’s instructions. For detailed description see the appendix.We enriched protein-coding DNA using Human Whole Exome kit v7  according to the manufacturer´s instructions. For detailed description see the appendix.The data curation was performed using the standard tools – FastQC, MultiQC  for qualDNA methylation analyses were done by the Infinium Methylation EPIC Kit  according to the manufacturer’s protocol and as described in Honkova et al. Raw data were processed with the minfi package . Data weRNA isolation, quality and quantity analyses were all performed according to the manufacturer’s instructions. For detailed description see the appendix.Ribosomal RNAs (rRNAs) were removed using NEBNext rRNA Depletion Kit . RNA sequencing cDNA libraries were constructed according to the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina, as provided by the manufacturer . For detailed description of sequencing see the appendix.The data curation was performed using the standard tools – FastQC, MultiQC for quality control , TrimmomRNA isolation, quality and quantity analyses were all performed according to the manufacturer’s instructions. For detailed description see the appendix.The next-generation sequencing library preparation was carried out as described in Sabo et al.  and CervFastQC and MultiQC  were useFor external validation of our data, the GSE62321 set was used , 49. ThiIPO5 gene interactions, the correlation data were downloaded from the CRISPR (Avana) Public Depmap v20Q3 portal  for all cell lines in the database . Gene correlation between IPO5 knockout effect and gene expression was considered significant for a p-value lower than 0.05 and an absolute value of correlation coefficient higher than 0.1. Functional enrichment analysis of significantly correlated genes with the IPO5 effect was performed using the clusterProfiler, and ReactomePA R packages. Only terms associated with a BH-adjusted p-value lower than 0.05 were considered.To analyze the The clinical and pathological characteristics of 10 patients with liver metastasis included in the study are described in We have successfully analyzed the DNA isolated from 10 paired CRCLM and adjacent liver tissue samples.After alignment, a range from 81 to 169 million properly aligned read pairs per sample were obtained. In summary, the percentage of adequately mapped read pairs reached 99% for all samples. The average read depth for each patient can be found in With the advantage of robust WES data, several features, such as mutational analysis, tumor mutation burden and copy number variations were analyzed with obtained WES data. The first analysis of all samples was performed, and then stratification for either SmCRC or MmCRC was presented.WES in CRCLM and adjacent liver tissue DNA was performed to identify somatic tumor-specific single nucleotide variants (SNV) and short insertion/deletion (Indel) mutations characterizing liver metastasis.In total, 1,900 heterozygous and homozygous SNVs and Indels  have been discovered as different between these two groups: 104 with high, 361 with moderate, 645 with low and 816 with modifier putative effect. Regarding the SNV effect, the largest group of identified variants belonged to the missense and intron variants, followed by synonymous variants. For the Polyphen2 calculation, we filtered out only non-synonymous exonic variants, and the results were as follows: benign prediction was among all samples in the range 32-48%, possibly damaging in the range 17-33%, and damaging prediction in the range 28-48%. After stratification for chromosome localizations, the observed variants’ distribution was similar across all chromosomes (APC (75%), SYNE1 (50%), TP53 (50%), and TTN (50%) genes. Only one APC mutation  was detected more than once  and MmCRC (n=3) patients. No significant differences in distribution among these two groups were noticed after stratification for putative effect, SNV effect, chromosome distribution, or Polyphen2 characterization  and USH2A, ACACA, ALS2CL, ATP10A, BRINP2, CACNA1l, CCNJ, CFAP46, DYNC2H1, ENPP7P13, KIA1549L, MED13L, PCDHA10, PRPF6, PTPRT (0% vs 66%) genes. No other substantial differences were observed between these two groups , multiple nucleotide variations (MNVs), and InDels per Mb was calculated. TMB was defined as aTMB, which includes synonymous mutations, and fTMB, which excludes synonymous mutations. The average value of aTMB was 5.99 in all patients, and the median value of aTMB was 6.62 in all patients . The average value of fTMB was 5.21 in all patients, and the median value of aTMB was 5.75 in all patients .Mutational signature analysis revealed that transition mutations were more common than transversion mutations, and C > T substitutions were predominant in our samples  SNVs by analyzing the six mutation classes . According to the COSMIC SBS V3 database, we identified eight mutational signatures with a higher than 5% proportion in our CRCLM samples  and SBS6 (cause is related to defective mismatch repair) mutational signatures, while MmCRC patients displayed a more heterogeneous distribution of mutation signatures. Given the small number of patients in each group taking this result with caution is necessary.Using PureCN, we identified recurrent CNVs affecting 5,432 genes  bases while the median deletion length was 116,0517 . For MmCRC, median duplication length was 3,267,214 and median deletion length 607,852 . Hypermethylated CpG sites were predominantly observed in CpG islands (64.4%). On the other hand, hypomethylated CpG sites were mainly located in intergenic regions, the so-called open sea (79.8%)  were identified by an analysis of CRCLM compared with colon tissue. Of these, 1,719 were up-regulated, and 992 were down-regulated in CRCLM samples  uncovered 124 significantly deregulated pathways in the Reactome terms .In the terms of chronicity, only two genes were significantly down-regulated in SmCRC compared to MmCRC . In total, 1,603 miRNAs were detected, with an average of 845 miRNAs detected in each sample and 670 miRNAs with sufficient coverage across all samples for differential expression calculation. Other classes of sncRNAs were seen, including, tRNAs, and snoRNAs. The distribution of the detected non-miRNA sncRNAs in the sample group is shown in After sequencing, an average of 57.3% of the reads were aligned to miRNA sequences, while 10.2% were aligned to other small non-coding RNAs (sncRNAs)  Public Depmap v20Q3 portal  for all cell lines in the database . With the threshold for adjusted p-value 0.05 and absolute value of correlation coefficient >0.1, 1,739 genes that interact with the IPO5 gene were identified.To analyze the IPO5 gene enriched in Reactome were related to the cell cycle. The Reactome analysis for IPO5 gene was visualized in The most significant results obtained for It is well known that one miRNA can regulate several genes and vice versa, multiple miRNAs can regulate one gene. For this reason, we were also interested in overlapped genes without considering the miRNA expression levels. This combined analysis resulted in 107 genes . Comparing this set with our set, the intersection resulted in 9 significant DEGs  that was detected more than once. No other mutation was observed in any other gene more than once. Similarly, Naxerova and co-authors reported that in 65% of cases, regional lymphatic and distant metastases arose from independent subclones from the primary tumor  regulation . The data presented here also provide a valuable resource for understanding molecular distinctions between SmCRC and MmCRC, and have the potential to enhance the diagnosis, prognostication and management of CRCLMs by a molecularly targeted approach. The strength of this study lies in the sizeable high-throughput analysis, including the non-trial patients reflecting real-world patients. On the other hand, we are aware of certain limitations of our study, such as the lack of primary tumors due to the prospective design. However, the intersection between our and validation sets confirmed the validity of our results.In summary, we have identified genes and pathways that may be considered actionable targets in CRCLMs  and the Institute of Experimental Medicine, Prague, Czech Republic . The patients/participants provided their written informed consent to participate in this study.VV, JP, and LV secured the financial funding, VV and SF were responsible for the experimental design, OK, SJ, MM, and JP collected the patients’ samples, MK was responsible for the pathological assessment of the samples, AS performed the isolation of the samples and preparation for sequencing methods, KH and IC performed the methylation array analyses, JH and SG-M were responsible for bioinformatical analyses of data from all methods, VV, JH, and OK wrote the paper with the input from all authors. All co-authors have participated in the planning and execution of the study as well as the preparation of the manuscript. All authors contributed to the article and approved the submitted version.


The values of the carbon sequestration rate were 0.15, 0.13, 0.13, and 0.09 Mg C ha−1 year−1 in RG5, PG, CG, and SPG management, respectively. This indicates that rotational grazing management might be the optimal choice for improving vegetation carbon accumulation in sparse woodlands. This study contributes to decision-making on how to choose livestock grazing management to maintain higher carbon storage.Livestock grazing is a widespread practice in human activities worldwide. However, the effects of livestock grazing management on vegetation carbon storage have not been thoroughly evaluated. In this study, we used the system dynamic approach to simulate the effects of different livestock grazing management strategies on carbon stock in sparse elm woodlands. The livestock grazing management strategies included rotational grazing every 5 years (RG5), prohibited grazing (PG), seasonal prohibited grazing (SPG), and continuous grazing (CG). We evaluated the carbon sequestration rate in vegetation using logistical models. The results showed that the carbon stock of elm trees in sparse woodlands was 5–15 M g ha This reoodlands . Besidesla trees .U. pumila life cycle, i.e., seed (i = 1), seedling, juvenile, mature, and over-mature (i = 5) tree stages. In this model, the seed stage is linked to the mature and over-mature stages, as seeds are produced in these two stages. The seeds become seedlings at a specific germination rate. U. pumila trees die at a stage-specific rate, except for the seed stage. At the over-mature stage, no transition is made to the next stage; as a result, the over-mature stage is the last in the U. pumila life cycle. The equations depicting the elm life cycle are shown below . WC represents the water consumption per individual at each of the developmental stages, except for the seed stage. P represents precipitation, which follows a normal distribution, whose parameters, i.e., mean and variance, were calculated according to long-term collected data. Besides the parameters mentioned above, other parameters include death rate, seed germination rate, and the period of each morphological stage. The values of all parameters used in the system dynamic model are shown in Here, N represents the number of individuals at each of the five stages, while B and D represent birth and death rates, respectively. TP stands for the transition probability  was obtained with a logistic model, where the carbon sequestration rate serves as a parameter and is estimated using 0 is the initial carbon stock at age 0, representing the amount of carbon present in the vegetation at the time of planting or the start of observation. Rseq is the C sequestration rate, which governs the rate at which the vegetation assimilates atmospheric carbon. Here K, N0 and Rseq are parameters needed to estimate.In i.e., rotational grazing every 5 years (RG5), prohibited grazing (PG), seasonal prohibited grazing , and continuous grazing (CG). Continuous grazing increases elm seed production and decreases elm seedling densities in sparse elm woodlands in a previous study . A unit-consistency test, which checks for agreement among units, was used to validate this model. It was automatically completed in the Vensim package .P < 0.05). The parameters were significant at a critical value (P < 0.05). The standard error (SE) of the estimated parameters and residual standard error (RSE) of the logistical model were reported.The models in In the first 10 years, the carbon stock of elm trees was almost the same under all four management techniques. From the 10th year to a 100 years, the carbon stock of elm trees in RG5 and PG management was higher than in CG and SPG management. From the 20th year to a 100 years, the carbon stock of elm trees under CG management was more substantial than under SPG management.When comparing RG5 to PG management, the carbon accumulation in the PG treatment surpassed that of the RG-5 between the 70th and 80th years, but was diminished between the 90th and 100th years. In contrast, when comparing RG5 with CG management, carbon sequestration in CG was inferior to that in RG-5.From the beginning to the 80th year, the ability to store carbon increased under RG5, PG, CG, and SPG management. After the peak, the ability to maintain carbon stock decreased under RG5, PG, and CG management, while the ability to maintain carbon stock remained steady under SPG management .−1 throughout the simulation period under SPG management. In contrast, the carbon stock of elm trees exceeded 5 M g ha−1 almost from the 15th year and onwards under CG management. Additionally, the carbon stock of elm trees exceeded 5 M g ha−1 after the 20th year under RG5 and PG management. Furthermore, the carbon stock of elm trees under RG5 and PG management ranged between 10 and 15 M g haz−1 after the 40th year  was notably higher under PG management. This could account for the observed higher carbon stock in PG as compared to CG throughout the duration of the study. This data suggests a hierarchical efficiency in carbon sequestration by elm trees under different management practices: RG5 being the most efficient, followed by PG, CG, and then SPG.Elm trees exhibited varying carbon sequestration rates under different management practices: 0.15, 0.13, 0.13 and 0.09 Mg C haManagement practices can significantly shape vegetation carbon sequestration rates by influencing the structural and functional attributes of plant communities . For insConversely, mismanaged practices can have deleterious effects on vegetation carbon sequestration. Overgrazing, for example, can result in the dominance of less productive plant species, diminishing the overall carbon capture capacity of the vegetation . Practic−1 year−1 for trees within the Ulmaceae family. This further attests to the appropriateness of employing logistic models to capture the accumulation dynamics of biomass in vegetation. It is worth noting, however, that the carbon sequestration rates derived from our logistic models represent an average value across the entire life cycle of the vegetation and do not imply that the annual carbon sequestration rates for the plants consistently maintain this state.Our findings, indicating carbon sequestration rates for elm trees ranging between 0.09 and 0.15 under various management measures, align with those reported by In addition to grazing management, the duration of grazing and grazing exclusion management can also influence carbon storage in soil and vegetation . For exaClimate changes, especially changes in precipitation and temperature, can affect vegetation carbon stock as biomass accumulation is influenced . PrecipiLivestock grazing can decrease the herb layer in terms of biomass, coverage, and spatial distribution . SubsequIn our research, we crafted an advanced system dynamics model that integrates multiple life-history stages to explore the impact of livestock grazing management strategies on carbon storage, both in terms of carbon stock and sequestration rates. Importantly, for a nuanced estimation of carbon sequestration rates, we employed logistical models.−1 year−1.In this study, we examined four distinct grazing management strategies: RG5, PG, CG, and SPG. Our findings indicate that the carbon stock of elm trees under RG5 and PG management surpassed those observed under CG and SPG management. Moreover, our research highlights the differences in carbon sequestration rates of elm trees across these managements, with RG5 standing out as the most effective. As a result, our data suggests that rotational grazing management may offer a superior strategy to enhance vegetation carbon storage in sparse elm woodlands in semi-arid landscapes. Furthermore, in line with previous research, our logistic model outcomes showed that the carbon sequestration rates for elm trees ranged between 0.09 and 0.15 Mg C haOur assessment sheds light on the carbon sequestration capabilities of vegetation in sparse elm woodlands, offering valuable insights for grazing management in semi-arid regions. For a more refined understanding of carbon storage capacities in the future, it will be imperative to integrate soil carbon sequestration studies and delve deeper into the interactions between the herb layers and elm trees within these sparse woodlands.