Coronary artery disease (CAD), the leading cause of death worldwide, is caused by atherosclerosis. Atherosclerosis has an inflammatory component, which can be measured with the neutrophil-to-lymphocyte ratio (NLR). Vitamin D has anti-inflammatory and anti-atherogenic properties that affect many mechanisms involved in CAD. In this study, we investigated the association between NLR, vitamin D levels, and the severity of CAD in a group of patients who had a myocardial infarction (MI) in the past. Our results show that patients with acute coronary syndrome had a higher NLR compared to those with stable CAD. No associations were observed between NLR and the severity of CAD. We found no correlation between vitamin D levels and NLR. NLR could be used as a prognostic marker of consecutive MI in patients with CAD and previous MI.p < 0.05). No associations between NLR and severity of CAD (p = 0.14) in the cohort and in the subgroups with stable CAD (p = 0.40) and ACS (p = 0.34) were observed. We found no correlation between vitamin D level and NLR neither in the whole study group (p = 0.29) nor in subgroups of patients with stable CAD (p = 0.84) and ACS (p = 0.30). NLR could be used as prognostic biomarker of consecutive MI in patients with CAD and a history of MI.Coronary artery disease (CAD), the leading cause of death worldwide, has an underlying cause in atherosclerosis. The activity of this inflammatory process can be measured with neutrophil-to-lymphocyte ratio (NLR). The anti-inflammatory and anti-atherogenic properties of vitamin D affect many mechanisms involved in CAD. In this study, we investigated the association between NLR, vitamin D concentration, and severity of CAD in a group of patients with a history of myocardial infarction (MI). NLR was higher in patients with acute coronary syndrome (ACS) in comparison to those with stable CAD (median: 2.8, range: 0.96–24.3 vs. median: 2.3, range: 0.03–31.6;  Coronary Artery Disease (CAD) is the leading cause of death worldwide. As stated by the World Health Organization (WHO), CAD was responsible for 16% of all deaths in the world in 2019, which results in it being the second greatest cause of disability-adjusted life years  but alsoWhite blood cell count, a widely available inflammatory marker, was previously associated with a greater extent of CAD and increased mortality in patients with unstable angina or non-ST segment elevation myocardial infarction (MI) . As lympVitamin D has diverse properties, among which its anti-inflammatory and anti-atherogenic effects are of particular interest . It modiAlthough the current indications to supplement vitamin D in patients with CAD are not different than in the general population , there a3 µL, active neoplastic processes or paraneoplastic syndromes, elevated concentrations of inflammatory markers—C-reactive protein concentration >5 mg/L or total white blood count exceeding 10 × 103 cells/µL, chronic kidney disease (stages III–V), calcium and phosphorus metabolism disorders as well as vitamin D ingesting as a diet supplement or a medication.The study is based on data from 268 patients who were hospitalized in the Cardiology Department of Bielanski Hospital, Warsaw, Poland, and underwent diagnostic catheter angiography to evaluate the extent of CAD between 2013 and 2017. The final statistical analysis included 181 men and 87 women, whose ages ranged from 36 to 93 years, with a history of a previous myocardial infarction treated with acetylsalicylic acid and atorvastatin or rosuvastatin. Details on clinical characteristics of the patients were presented in our previous articles ,40,41,42The patients enrolled in this study were measured in weight and height to diagnose obesity or overweight according to WHO criteria . Based o® 25 OH Vitamin D TOTAL Assay . It utilizes chemiluminescent immunoassay method, achieving a detection range between 4 and 150 ng/mL, with a precision equal to 5.0% of the coefficient of variation (CV), and a standard deviation of accuracy equaling 1.2% [® 25 OH Vitamin D TOTAL Assay were measured in ng/mL, where 1 ng/mL equals 2.5 nmol/L [Vitamin D serum concentrations were measured with the DiaSorin LIAISONing 1.2% . This teing 1.2% ,49,50. T5 nmol/L .The results of the vitamin D concentration were classified according to the Endocrine Society’s guidelines of clinical practice for vitamin D deficiency: concentrations <10 ng/mL, between ≥10 and <20 ng/mL, between ≥20 and <30 ng/mL, and ≥30 mg/mL were labeled severe deficiency, moderate deficiency, mild deficiency, and optimal concentration, respectively . As patiTotal blood count was measured in blood samples collected from the cephalic vein in di/tripotassium EDTA tubes by an automatic blood counter from this brand within two hours after the venepuncture. The NLR factor was defined as the quotient of absolute neutrophil count to absolute lymphocyte count.Coronary angiography, which uses X-rays and iodinated contrast to visualize coronary arteries, is the default method for stenosis assessment in CAD . In thist-test. Differences between prevalence in selected groups were determined with a Pearson’s chi-square test or a Fisher’s exact test. The NLR was compared between patients with different CASS scores using Kruskal–Wallis analysis. The relationship between the selected variables was analyzed with a Spearman’s correlation coefficient (R). Statistical significance was recognized if a two-sided p-value <0.05. Statistical analysis was completed with Statistica 13  and the figures were drawn with GraphPad Prism 8.0 .The data distribution were determined with a Shapiro–Wilk test. Comparison of continuous variables in the results was evaluated with a Mann–Whitney test or a Details on the study group are presented in p < 0.05; There was a significant difference in NLR between patients with stable CAD and patients with ACS  or ACS . Results are presented in There was no association between NLR and severity of CAD in the whole group D and NLR in the whole group , the study did not include other commonly used indicators such as interleukin 6 or ferritin.Chronic inflammation, including a cellular immune response, plays a key role in the pathogenesis of atherosclerosis and its complications . Recent We suggest considering carrying out well-designed studies to determine the role of the NLR indicator in predicting another infarction in patients with CAD diagnosis and a history of MI. The relevance of serum vitamin D concentration in patients with consecutive MI, as well as its influence on immune response and inflammatory markers also needs further well-designed research.In a group of patients with recurrent MI there is an elevation of NLR. Although no correlation was found between serum vitamin D concentration and NLR, the lowest vitamin D concentrations and the highest NLR were observed in patients with ACS. The role of vitamin D and NLR in CAD pathogenesis and its complications needs further research.


Subclinical atherosclerosis characterizes cardiovascular diseases (CVD), and Human Immunodeficiency Virus (HIV) infection and antiretroviral therapy (ART) are identified risk factors for atherosclerosis. Meanwhile, data on HIV and atherosclerosis in Nigeria are limited.We sought to estimate the prevalence of subclinical atherosclerosis and associated risk factors amongst adult persons living with HIV/AIDS (PLHIV) enrolled at University of Abuja Teaching Hospital, Gwagwalada, Abuja (UATH).This was a cross-sectional study of 277 consecutively selected PLHIV ≥18 years enrolled for HIV care and treatment at UATH. Pretested structured questionnaire was used to collect data from consenting ART-experienced and ART-naïve patients on risk factors of atherosclerosis. Carotid intima media thickness (CIMT) ≥0.71 mm as measured by Doppler ultrasonography was used to identify patients with sub-clinical atherosclerosis. Two logistic regression models with (Model-A) and without (Model-B) traditional risk factors were fitted to identify risk factors of subclinical atherosclerosis.p = 0.002], advancing age ; ≥40 years AOR 19.51(4.30–88.56), p ≤ 0.001), advancing HIV infection , hypercholesterolemia  and ≥5 year duration on ART  as risk factors of subclinical atherosclerosis. In Model-B  on the other hand, advancing HIV infection  and duration on ART  were found as risk factors of subclinical atherosclerosis.Participants' mean age was 39.44 ± 10.71 years with female preponderance (64.26%). Overall prevalence of subclinical atherosclerosis was 43.32% (62.25% in ART-experienced). Model-A identified male sex [AOR 4.33(1.74–10.76), Subclinical atherosclerosis was higher in ART-experienced patients, and this was irrespective of presence or absence of traditional risk factors. And advancing HIV disease and duration on ART were found as significant risk factors for subclinical atherosclerosis. We therefore recommend routine CVD risk screening in PLHIV. Human Immunodeficiency Virus (HIV) infected individuals appear to have significantly higher risk of myocardial infarction and coronary heart disease relative to HIV negative individuals –4. With Meanwhile, the presence of atherosclerosis can be diagnosed early at subclinical levels using carotid intima media thickness (CIMT) as a measure –23, and A Ugandan study to estimate the prevalence of subclinical atherosclerosis among HIV infected adults using a CIMT of ≥ 0.78 mm as cut off found an 18% prevalence . HoweverWe conducted a cross-sectional study amongst HIV infected adults 18 years and above at the University of Abuja Teaching Hospital, Gwagwalada (UATH). Our study population comprised both ART experienced patients with at least six months of ART as well as ART naïve patients who were newly enrolled into care.UATH is a 350-bedded tertiary facility located in Gwagwalada in the Federal Capital Territory in Nigeria. UATH runs clinic for persons infected with HIV with about 4,864 patients currently receiving life-long ART as of December 2018 , and an average monthly ART enrollment rate of about 30 patients per month.HIV testing services at UATH are routinely provided at multiple service delivery points including the general outpatient department, inpatient wards, the main laboratory, antenatal clinic, labour ward, TB clinic, family planning clinic, and sexually transmitted infection clinic using serial algorithm based on the Nigerian guidelines . PatientUsing Kish and Leslie's formula for prevalence studies, and assuming a prevalence of 18%  for subcEligible patients enrolled into HIV care and treatment who consented to take part in the study were consecutively enrolled into the study until the required sample size was achieved for each group. Persons with history of cardiovascular risk including hypertension, diabetes, peripheral vascular disorder, myocardial infarction, heart failure, or stroke, who are likely to have clinical evidence of atherosclerosis were excluded from the study. Other exclusion criteria include persons with life-threatening opportunistic infection requiring critical care, or cognitive impairment precluding the patient from providing appropriate responses to interview questions or providing consent.A structured data collection tool divided into four sections was used for data collection. These sections are: (1) screening questions to rule out study exclusion criteria, (2) sociodemographic and HIV related clinical characteristics of participants, (3) physical examination findings, and (4) laboratory and radiology results. The data collection tool was pretested and validated with 10 patients at UATH preceding the study. Pretest data were excluded from the analysis.The primary outcome measure for this study was the presence of subclinical atherosclerosis amongst PLHIV enrolled in this study. Subclinical atherosclerosis was defined as carotid intima media thickness ≥0.71 mm as measured by Doppler ultrasonography . InformePhysical examination was carried out to check for signs and symptoms of opportunistic infections with classification of participant into applicable WHO clinical stages as elucidated in the WHO clinical staging criteria , 36. BloCarotid Doppler ultrasonography was performed using a high resolution 10 MHz linear array transducer of LOGIC F series GE ultrasound machine  by a trained cardiologist. Participants were requested to remove jewelry around the neck. Ultrasound of the carotid intima media was performed with the subject lying supine on the couch to the right of the examiner with pillow support under the neck to achieve the desired neck extension and head turned 45° away from the side being scanned. Adequate amount of coupling gel was applied to the scan area to eliminate air gap between probe and skin surface. Right and left CCA were located by longitudinal and transverse scans. Three measurements of the CIMT were obtained at the far wall on each side, each at 1 cm proximal to the right and left carotid bulb. The mean of three point measurements each of CIMT thickness for both right and left CCA were calculated and used for the analysis.Laboratory request forms carrying the participant's study identification number were given to the patient and requested to return the following day for laboratory tests after an overnight fast of 8 to 12 h. On return, about 10 mls of venous blood sample was obtained under aseptic conditions. One drop of blood was used to test for fasting blood sugar using an Accucheck glucometer, while the rest of the sample was divided in aliquots of about 5 mls into vacutainer tube containing ethylene diamine tetra-acetic acid (EDTA) anticoagulant by Becton Dickinson, and plain bottle, for viral load testing and lipid profile, respectively. The blood was centrifuged for 10 min at 3,000 rpm within 2–4 h of collection, and the separated plasma and serum respectively were stored at −20°C. Lipid profile analysis was done using Landwind C100 Plus Chemistry Autoanalyzer for the total cholesterol (TC), High density lipoprotein-cholesterol (HDL-C), and triglycerides (TG). Low density lipoprotein-cholesterol (LDL-C) was calculated from TC, HDL-C and TG using Friedewald formulae (LDL-C = TC-TG/ 5 + HDL-C) , 41.On tExplanatory variables included age, sex, history and duration of smoking habits, history of use of alcohol, systolic blood pressure, diastolic blood pressure, fasting blood sugar, serum lipid profiles , body mass index, waist circumference, history of ARV use, duration on ART, ART regimen-line and class of ARV used. All completed data collection tools as well as all laboratory and radiological results were retrieved and duly entered into Statistical Package for Social Sciences (SPSS) software version 21.0 for storage and analysis .2 (underweight), 18.5–24.99 Kg/m2 , 25–29.99 Kg/m2 (overweight) or >30 Kg/m2 (obese)  . Normal 0 mmol/L . Fasting0 mmol/L . Viral lp-value of <=0.2 in the bivariate analysis were included in the multivariate model to have a robust mix of predictors to consider. Two multivariable logistic regression models were fitted to explain on the one hand all possible risk factors for atherosclerosis (Model A), while a second model sought to explain the role of HIV infection and ART on atherosclerosis ((Model B). All statistically significant variables from the bivariate analysis were fitted into the multivariable logistic regression model in a stepwise manner to identify all possible risk factors for atherosclerosis (Model A). A second multivariable logistic regression model excluded all the modifiable traditional risk factors of atherosclerosis such as smoking, diabetes mellitus, dyslipidemias, hypertension, and obesity (Model B). A p-value <=0.05 was considered statistically significant for all multivariable analysis.Bivariate analysis using Chi-square test was used to determine factors associated with subclinical atherosclerosis. Factors with a The study protocol was submitted to the human research and ethics committee of UATH for ethical review and approval.p < 0.001). Prevalence of subclinical atherosclerosis increased with age, p < 0.001. Prevalence of subclinical atherosclerosis was higher among participants who reported no alcohol use than those who reported alcohol use .A total of 120 (43.30%) study participants 95% CI: (37.45–49.19) had subclinical atherosclerosis. Subclinical atherosclerosis was more prevalent in males than females . Subclinical atherosclerosis was more prevalent in study participants with total cholesterol ≥200 mg/dl than those with total cholesterol <200 mg/dl . Prevalence of subclinical atherosclerosis was higher among study participants with low density lipoprotein >130 mg/dl than those with low density lipoprotein <130 mg/dl . Subclinical atherosclerosis was more prevalent in study participants with triglycerides ≥150 mg/dl than those with triglycerides <150 mg/dl . Body mass index, diastolic blood pressure, fasting blood sugar, high density lipoprotein, and waist-hip circumference ratio were not associated with subclinical atherosclerosis.Prevalence of subclinical atherosclerosis was higher among study participants with systolic blood pressure ≥140 mm Hg than those with systolic blood pressure <140 mm Hg . Of the 151 ART experienced patients studied, the prevalence of subclinical atherosclerosis increased with increase in the duration of usage ART with a prevalence of 22.20% in those with <2 years usage, 41.20% in those with 2–4 years usage, and 72.20% with ≥5 years usage: p < 0.001). 60% of the patients on PI, 47.06% on NNRTI, 38.57% on Integrase inhibitor had subclinical atherosclerosis (p = 0.017). There were no significant associations between subclinical atherosclerosis and baseline CD4, WHO staging at ART initiation, WHO staging at study enrollment, current ART regimen-line, and NRTI backbone or viral load.ART status was associated with subclinical atherosclerosis with 26 (20.60%) of ART naive and 94 (62.30%) of ART experienced patients having subclinical atherosclerosis  region (0.72 mm) and increased with age, with mean CIMT of 0.59 mm, 0.63 mm, 0.76 mm and 0.83 mm in age groups 40–49 years, 50–59 years, 60–69 years and >70 years respectively . Our thrIn the bivariate analysis, subclinical atherosclerosis was associated with exposure to ART with a higher prevalence compared to ART naïve patients. This finding is comparable to findings by Sarfo et al. , in GhanIn model A, after adjusting for confounders the only significant risk factors identified from our study for subclinical atherosclerosis included male sex, age of 40 years and above, advancing HIV infection (WHO stage II & above at study enrollment) and duration on ART of five years or more. Male participants had four-fold increase in the risk of having subclinical atherosclerosis compared to females similar to the findings of Albuquerque et al.  who repoOur study provides better understanding of the prevalence and predictors of subclinical atherosclerosis in HIV positive patients in our environment to inform prevention and management of CVD in PLHIV. However, our study has a few limitations. We employed a cross-sectional study design. A longitudinal or cohort design would have been more appropriate to firmly establish the temporal effect of HIV or ART on the aetiopathogenesis of subclinical atherosclerosis. Another limitation of our study was the lack of longitudinal data on regimen changes (substitutions and switches) as well as duration on specific regimen which would have enabled us to further profile the cardiotoxic effects of specific ARV molecules in the aetiopathogenesis of subclinical atherosclerosis. Furthermore, a lack of data on date of HIV diagnosis for all patients or longitudinal data on viral load assays limits our ability to explore the role of prolonged viraemia in the aetiopathogenesis of CVD. We adopted a cross-sectional design due to funding limitations. The adopted methodology was however sufficient to answer our study questions. Future studies may consider exploring these outstanding questions.We found a high prevalence of subclinical atherosclerosis amongst ART experienced persons in our study. Associated risk factors for subclinical atherosclerosis include male sex, advancing age, hypercholesterolaemia, advancing HIV disease, and duration on ART. Even in the absence of traditional risk factors for CVD, advancing HIV infection and duration on ART are significant risk factors for subclinical atherosclerosis.Our study findings strengthen the case for cardiovascular risk screening amongst people living with HIV.


The human genome presents variation at distinct levels, copy number variants (CNVs) are DNA segments of variable lengths that range from several base pairs to megabases and are present at a variable number of copies in human genomes. Common CNVs have no apparent influence on the phenotype; however, some rare CNVs have been associated with phenotypic traits, depending on their size and gene content. CNVs are detected by microarrays of different densities and are generally visualized, and their frequencies analysed using the HapMap as default reference population. Nevertheless, this default reference is inadequate when the samples analysed are from people from Mexico, since population with a Hispanic genetic background are minimally represented. In this work, we describe the variation in the frequencies of four common CNVs in Mexican-Mestizo individuals.In a cohort of 147 unrelated Mexican-Mestizo individuals, we found that the common CNVs 2p11.2 (99.6%), 8p11.22 (54.5%), 14q32.33 (100%), and 15q11.2 (71.1%) appeared with unexpectedly high frequencies when contrasted with the HapMap reference (ChAS). Yet, while when comparing to an ethnically related reference population, these differences were significantly reduced or even disappeared.The findings in this work contribute to (1) a better description of the CNVs characteristics of the Mexican Mestizo population and enhance the knowledge of genome variation in different ethnic groups. (2) emphasize the importance of contrasting CNVs identified in studied individuals against a reference group that—as best as possible—share the same ethnicity while keeping this relevant information in mind when conducting CNV studies at the population or clinical level. In 2004, two independent groups, Iafrate and Sebat [The human genome presents distinct variations: single nucleotide variants, insertion‒deletion of a few nucleotides, repetitive sequences of a variable number of nucleotides, and structural variants. The human genome is 3.1 giga base pairs (Gb) in size, distributed in 23 pairs of chromosomes so that every individual inherits one copy from each parent and has two copies or copy number (CN) CN = 2 of every nd Sebat , 6. In snd Sebat , 8. CNV nd Sebat –11. A CNnd Sebat . Neverthnd Sebat .The accurate detection and interpretation of CNVs are essential for research and clinical diagnostics because CNVs may be a general population variant or associated with pathology . TherefoThe Database of Genomic Variants (DGV) was initiated to provide a publicly accessible, comprehensive, and curated catalog of genomic variants . The DGVIn general, this could represent an issue since the contribution of the ethnic group is diluted when the entire group is used as a reference.Therefore, it is necessary to report information on this type, especially for underrepresented populations.Several methodologies are commonly used for CNV genome-wide study; one of the most robust methods is microarray analysis, mainly when an array contains a high density of probes. Most CNV studies are performed with single nucleotide polymorphism (SNP) microarrays, which, in addition to providing information on CNV in the genome, also provide information at the allele level, highlighting the presence of regions/runs of homozygosity called long contiguous stretches of homozygosity (LCSH) and providing essential clues regarding parental relatedness (consanguinity), uniparental disomy, chromosomal recombination or rearrangements , 21.This work describes the frequency of four common CNVs found using SNP microarrays in unrelated Mexican-Mestizo individuals, who were healthy or had aneuploidies, and in whom structural alterations in their genome was not suspected. The study highlights the frequency variation of these common CNVs depending on the ethnic origin of the population used as reference for comparison.It is essential to recognize the ethnic differences in the distribution and frequency of CNVs, not only to recognize their contribution to structural chromosomal variation but also to ease recognition of possible associations between CNVs and phenotypic characteristics that may or may not be pathogenic.We studied the presence of CNV in a population of 147 Mexican-Mestizo individuals, 50 phenotypically normal  and 97 patients with aneuploidies , which are part of a more extensive study that aims to find genomic differences among these groups. In this first study, we show four CNVs in common among all these groups, with a very high frequency in our population when contrasted with distant populations, as is the mix of the HapMap reference that includes African, Asian, and Caucasian individuals.To recognize the presence of inbreeding and consanguinity in our sample population, we analyzed the regions with homozygosity or long contiguous stretches of homozygosity (LCSH) with Affymetrix Genome-Wide Human SNP Array 6.0 (GW SNP 6.0). Eighteen of the 50 healthy controls (MXM) studied showed at least one autosomal region with LCSH. In the aneuploidy group (MXM_A), 73/97 presented at least one autosomal region with LCSH.In addition, we analyzed 99 CEL files from 99 blood samples of Mexican women with breast cancer (MXM_WBC) present in public databases, where 75/99 presented at least one autosomal region with LCSH.Neither of these LCSH regions overlaps with the CNV reported in this study, and more than 42% are common LCSHs, such as the 2q11.1q11.2, 11p11.2p12.1, 16p11.2p11.1, and 20q11.21q11.23 regions previously reported –24. The We deduced the ancestry in our MXM and MXM_A groups compared with the Mexican ancestry in Los Angeles CA, USA (MXL) included in the HapMap project . First, we noticed that our population (MXM and MXM_A) has almost the same genetic content of European descent from Utah, USA (CEU), Native American (NAT), and Yoruba from Nigeria (YRI), then those studied for the HapMap, with NAT being the most significant contribution, even greater than for MXL, 0.488 vs 0.678 and 0.618, then of Caucasians and finally Africans ), 8 (0.15 Mb), 14 (1.02 Mb), and 15 (0.76 Mb) with a high frequency in the general population of Mexican-Mestizo (MXM), in Mexican-Mestizo with aneuploidy (MXM_A), and in the MXM_WBC group Figs.   was CN = 2  have null CN = 0. The frequency of CN ≠ 2 across all the groups is 54.5% . For chromosome 8, locus 8p11.22 A total of 14/16 (87.5%) samples showed < 2 copies. For 14q32.33, we corroborated the gain of the CNV in 13/16 (81.25%) with 3 to 5 copies; for chromosome 15, we found both losses and gains by GW, and we found 4 patients with loss and 6 with copy gains   in .75% in lThe CNV found with the Chromosome Analysis Suite (ChAS) was validated with an additional CNV calling method CRMA v2.We declare the raw data set conformed by 50 MXM CEL files (Genome-Wide Human SNP Array 6.0).vs MXM reference. B) C12 vs Spanish population (IBS) reference. C) C12 vs CEU reference. D) CN in the C12 vs. YRI reference. E) CN in C12 vs. two populations included in the HapMap reference (CEU + YRI). It is important to note that the difference in CN in our sample depends on the reference used.This method could compare the copy number estimates in 14q32.33 for each sample. Figure It is clear that in A and B, the 14q32.33 region does not show changes with respect to the Mexican and Spanish reference population, emphasizing our Mestizo race formed by Spanish and Native American groups. In contrast, comparisons with CEU, YRI and CEU + YRI make evident a gain of probes located in the chr14:106,078,230–107,100,266 region, showing our racial differences with these populations.loci: 2p11.2, 8p11.22, 14q32.33, and 15q11.2. No genes were found at 2p11.2 or 15q11.2, while the CNV at 8p11.22 included the genes ADAM5 (HGNC: 212) and ADAM3A (HGNC: 209). The 14q32.33 locus comprises the genes KIAA0125 (HGNC: 9834), ADAM6 (HGNC: 8755), LINC00226 , IGHD (HGNC:5480), IGHG3 (HGNC:5527), and IGHG1 (HGNC:5525); none of these genes has been associated with Mendelian diseases (OMIM) related to genetic dosage , and there are no regulatory sequences that could modify gene expression in any of the regions of the four CNV; neither of our CNV reported overlap with LCSH regions.In the tested samples, we found four high-frequency CNVs in the following http://cs-tl.de/DB/CA/sSMC/0-Start.html and the chromosomal heteromorphisms database http://cs-tl.de/DB/CA/HCM/0-Start.html, we found no evidence that a number other than CN = 2 in the 2p11.2, 8p11.22 and 15q11.2 regions or 14q32.33 duplications is associated with pathology, indicating that numerical variants in these regions have no impact on the phenotype, probably due to the absence of dosage-sensitive genes [Additionally, by searching these specific regions in databases such as the sSMC  database ve genes .The high frequency of some variants could be due to the existence of inbreeding or consanguinity in the population. However, according to our results on the percentage of homozygosity of the LCSH regions and the degree of inbreeding  , there iCNVs are genomic variants that confer high variability among individuals and have been recognized for several years –3. The fLater in 2015, the phase 3 1000 genome study included 2504 healthy individuals, only 64 of them  were individuals of Mexican descent, and they all were living in Los Angeles, California (MXL)  This stuNow we know that the CNV rate of occurrence has wide differences according to the ethnic group that is studied, as it occurs with other polymorphic traits such as SNPs or microsatellites . FurtherThe results presented here are based on the Affymetrix microarray platform and were analyzed using Affymetrix software, with Affymetrix annotations NetAffx 33.1, which is the default software provided with this commercial platform. This workflow is the one being used to find CNV for clinical and research studies in Mexico and in several other countries, however, in this software there is little representation of the Latin American population.locus is as compared to the default reference genome without taking into account possible ethnic differences of the studied population, an ethnic matched reference genome would not have shown gain in this region. Indeed in 2020, Godoy et al. identified a CNV obtained from three different microarray platforms from a Brazilian population to conform the Brazilian CNV database. They found that a 14q32.33 gain was present in 97.8% of the samples studied [In fact, there are studies reporting association of some of the polymorphisms studied, to pathological conditions –31; one The foregoing was addressed in this work by CNV calling (CRMA), where a Spanish population was used as reference, with this better-suited ethnical matched reference the gain of the CNV at 14q32.33 was reduced, and even did not appear as copy number gain , while when using an ethnically related population as a reference, the differences were reduced or disappeared, highlighting the importance of analyzing the CNVs of the studied individuals with a reference group that (as far as possible) shares the same ethnicity. These findings contribute to a better description of the CNVs characteristics of the Mexican-Mestizo population and enhance the knowledge of genome variation in different ethnic groups.We included blood samples from subjects with Mexican-Mestizo origin who consented to participating in this study. Fifty were healthy, unrelated volounteers from the general population. All subjects were over 18 years of age, had no history of genetic diseases and at the time of sample donation were not suffering from any infectious disease nor taking any medications . All participants had a normal karyotype 46,XX in females and 46,XY in males . We also included 97 aneuploid patients , with karyotype showing regular trisomy as well as X-monosomy without mosaicism, which was corroborated by interphase FISH .Genomic DNA was extracted from the blood samples obtained from the participants with the saline precipitation method .We analyzed the gDNA with the Affymetrix GeneChip® Genome-Wide Human SNP Array 6.0 . This array contains 906,600 SNP probes and 946,000 nonpolymorphic oligonucleotides; the median intermarker distance over all 1.8 million SNP and copy number markers combined is less than 700 bases.The procedures for DNA digestion, ligation, PCR amplification, fragmentation, labeling, denaturing and hybridization into the array were performed in 147 DNA samples  according to the protocols provided by the supplier. Arrays were stained and washed in the Affymetrix GeneChip Fluidic Station 450 and scanned using an Affymetrix GeneChip Scanner 3000 7G . We analyzed the files obtained with the appropriate bioinformatics tools.http://www.affymetrix.com). For LCSH > 3 Mb, the analysis configuration was set at LOH with marker count = 50 and size = 3000 kb, and for LCSH > 5 Mb, it was set at marker count = 50 and size = 5000 kb.We visualized the long contiguous stretches of homozygosity (LCSH) in Chromosome Analysis Suite (ChAS) software version 4.1, provided by Affymetrix . For the analysis, we used the NetAffx 33 hg19 annotation files  were estimated using LCSH > 3 Mb data; F was the total length of autosomal LCSH in kb divided by the total autosomal size covered by the Genome-Wide Human SNP Array 6.0 . We report the average of the F value for each group. An F value of 0.25 could reflect a first-degree parental relationship, 0.125 a second-degree relationship, 0.0625 a third-degree relationship and 0.03125 a fourth-degree relationship .To compare our studied population, we used the following CEL files obtained from Affymetrix GeneChip® Genome-Wide Human SNP Array 6.0 :(a) Ninety-nine files from healthy tissues obtained from women with breast cancer (MXM_WBC)  from theTogether with the MXM, they will be the Healthy tissues.(b) 30 Spanish (IBS) files downloaded from GEO data set GSE67047 (c) 30 Caucasian (CEU) and 10 Yoruba (YRI), part of the population studied in the HapMap project, CEL files kindly provided by Affymetrix/Thermo Fisher.The MAP and PED files obtained by Genotyping Console software (Affymetrix) were used for the ancestry analysis; we eliminated two files that did not pass quality control.The global ancestry of 49/50 healthy Mexican-Mestizo files (MXM) and 96/97 individuals with aneuploidy (MXM_A) was deduced through a supervised maximum likelihood ADMIXTURE approach from K = 2 to K = 3 ancestral components and compared with the global ancestry of the group MXL  (n = 64). For this analysis, genotypic frequencies of Northern European (CEU) (n = 99), Yoruba (YRI) (n = 108), and Native American (NAT) (n = 43), reported in the 1000 genomes project (1KGP), were considered as reference parental populations for demographic and historical reasons.CNVs were visualized with Affymetrix software . GeneChip Command Console (AGCC) software was used to generate the CEL files and ARR files from each microarray scanned. The CEL and ARR files were analyzed with Genotyping Console software  to obtain the CN data files (CNCHP) MAP and PED files for ancestry. CNCHP data were analyzed using Chromosome Analysis Suite (ChAS) software version 4.1 .http://www.affymetrix.com) has two ways of showing the results: one as a table that includes both coordinates and CN for each region; or a graphical interphase that displays the results of CN in an image of each of the 23 chromosomes, highlighting the regions with CN > 2 in blue arrowhead, regions with CN = 1 in red arrowhead, or CN regions with copy number (CN = 2), which are only marked in lines . The reporting threshold of the copy number was set at "High Resolution" settings, which gave us a total of gains or losses of 100 kb with a marker count ≥ 50. The copy number variants were compared among all samples. The CN analysis in ChAS software .For an additional CNV calling method, we used copy number estimation using robust multichip analysis, and this method provides full-resolution raw total copy number estimates by preprocessing and probe summarization. CRMA v2 is available in Bioconductor  and implBriefly, the CEL files were analyzed for quality after the program calibrated for crosstalk between allele probe pairs, followed by normalization for 25-mer nucleotide-position probe sequence effects. Next, we analyzed the performance of robust probe-summarization, normalized the PCR fragment-length effects on summary signals, and finally calculated the full-resolution total copy numbers. When a standard reference is used, it is often the average of a pool of samples CnChipEffectFile.The reference control samples for CRMA v2 analysis were a) the same 50 healthy individuals of MXM; b) 30 IBS, 30 CEU and 10 YRI. All groups were analyzed with the same pipeline.We made the following comparisons: (a) MXM vs. MXM reference; (b) MXM vs. IBS reference; (c) MXM vs. CEU reference; (d) MXM vs. YRI reference; and (e) MXM vs. HapMap reference (CEU + YRI).We reviewed publicly available databases to classify the CNVs found in our samples and the genes included:http://projects.tcag.ca/variation/).Database of Genomic Variants .DECIPHER (http://www.ecaruca.net/).ECARUCA (http://www.g2conline.org/).Genes2Cognition (https://www.ensembl.org/index.html).Ensembl (https://www.omim.org/).OMIM (http://geneontology.org/).Gene ontology (https://www.ncbi.nlm.nih.gov/clinvar/).ClinVar .ClinGen (http://cs-tl.de/DB/CA/sSMC/0-Start.html).The sSMC database (http://cs-tl.de/DB/CA/HCM/0-Start.html).The heteromorphims database (https://franklin.genoox.com/clinical-db/home


As an important nucleoside precursor in salvage synthesis pathway of uridine monophosphate, uridine (UR) is the most abundant nucleotide in sow milk. This study aimed to investigate the effects of maternal UR supplementation during second trimester of gestation on reproductive performance and amino acid metabolism of Sows.P < 0.05) with higher average piglet weight at birth in UR group (P = 0.083). Besides, dietary UR supplementation significantly increased TP in sow serum, BUN content in cord serum, and TP and ALB in newborn piglet serum (P < 0.05); but decreased AST level in sow serum and BUN level in piglet serum (P < 0.05). Importantly, free amino acids profile in sow serum newborn piglet serum and colostrum was changed by maternal UR supplementation during day 60 of pregnancy, as well as the expression of amino acids transporter (P < 0.05). In addition, from 100 to 2,000 μM UR can increased the viability of pTr2 cells. The UR exhibited higher distribution of G1/M phase of cell cycle at 400 μM compared with 0 μM, and reduced S-phases of cell cycle compared with 0 and 100μM (P < 0.05).Results showed that compared to CON group, the average number of stillborn piglets per litter was significantly reduced (Supplementation of uridine during day 60 of pregnancy can improve reproductive performance, regulate amino acid metabolism of sows and their offspring, and increase the viability of pTr2 cells. The gastrointestinal tract and visceral organ and skeletal muscle protein synthesis of piglet are influenced by sow milk . Interesin vivo, but the synthesis of endogenous nucleotides from amino acids requires a great deal of energy , which accounts for 86–98% of total 5′ nucleotides and has the highest content in sow’ colostrum . Therefof energy . It was A restricted feeding program is currently adopted by the swine industry to prevent excess weight gain during gestation, which can ease farrowing difficulties in gilts and sows, as well as appetite reduction of sows during lactation . HoweverTherefore, the objectives of the present study were to explore the effects of maternal UR supplementation on reproductive performance of sows, serum biochemical parameters, serum amino acids profile of sow and piglet, placental amino acids transporter, and the growth of porcine trophectoderm cell.The animal experiments were approved by the Institutional Animal Care and Use Committee of the Institute of Subtropical Agriculture, Chinese Academy of Sciences (2013020).60 multiparous crossbred sows (Landrace × Yorkshire) were selected according to parity (parity 4–6), expected delivery and body shape, and divided into two groups which fed a basal diet (CON) or a basal diet with UR supplementation 120 g/t) from 60th day of pregnancy to delivery. All sows were fed twice daily at 06:30 and 15:00 h and had free access to water during the experiment period. Each sow was fed 2.5–3.0 kg/d from days 60 to 107 of pregnancy and approximately 1.8 kg/d on day 5 before delivery. Farrowing was not induced and was constantly supervised. Sows were identified by ear tags and the reproductive performance of sow was recorded at delivery. Newborn piglets were individually weighed, and IUGR piglet weigh approximately 618–869 g, defined as weighing approximately 65% of the birth weight of the largest littermate in each litter  before eating colostrum from jugular vein , and cenn = 8 sows/group) in the similar place of placenta (great vessels were avoided) of piglets with BW about 1.5 kg were obtained immediately after farrowing. Part of placenta was stored in liquid nitrogen for RT-qPCR analyses  were randomly selected to collect colostrum (30–40 mL/sow) from the 4–6th mammary glands of each sow within 12 h postpartum by massaging breasts  was used for analyze the concentrations of serum triglycerides (TG), total cholesterol (TC), high density lipoprotein (HDL), and low-density lipoprotein (LDL) according to the commercial kits and manufacturer’s instructions which was purchased from Beijing Chemlin Biotech Co., Ltd. .The porcine trophectoderm cell line-2 (pTr2), which has been previously characterized and used for functional studies of porcine trophectoderm cell line, was originally isolated from the elongated porcine blastocysts, and collected on day 12 of pregnancy in this study . The celn = 6 wells/treatment). Cells were treated with 10% CCK-8 , diluted in normal culture medium at 37°C until visual color conversion occurred. OD (absorbance) values for each well were converted to relative cell number using standard curves.The pTr2 cells were subcultured in 96-well plates (30% confluence) in growth medium until the monolayer confluence reached 40% and then switched to custom medium free of serum and insulin. After starvation for 24 h, cells were added uridine  for 48 h to each well (n = 3 wells/treatment). The cultured pTr2 cells were trypsinized and fixed in 75% cold alcohol overnight. The fixed cells were stained by propidium iodide (PI), and analyzed by flow cytometer . ModiFit software was used to analyze the proportion of cells in G0/G1, S, or G2/M phase.The pTr2 cells were subcultured in 6-well plates (30% confluence) in medium until the monolayer confluence reached 40% and then switched to customized medium free of serum and insulin. After starvation for 24 h, cells were added uridine  for 48 h to each well .Total ribonucleic acid (RNA) was extracted from liver tissue using Trizol reagent . The total RNA was reverse transcribed into cDNA using reverse transcription kit from Takara Biomedical Technology, Japan. All primers were designed using Primer-BLAST on the National Center for Biotechnology Information (NCBI) website . Using St-test. In the cell experiment, Statistical significance was assessed by the one-way analysis of variance (ANOVA) followed by the Duncan test for multiple comparisons. Differences were considered statistically significant at P < 0.05. The IBM SPSS Statistics for Windows software, Version 22.0 , was used for statistical analyses.Data are expressed as the mean ± standard deviation of at least three independent experiments. The statistical significance between the two groups was assessed by the student’s P < 0.05). In addition, the average piglet weight at birth was higher in UR group than CON group (P = 0.083).The reproduction performance of the sows was summarized in P < 0.05), but decreased AST level (P < 0.05). The BUN content in cord serum was significantly higher by maternal UR supplementation (P < 0.05). Meanwhile, the serum TP and ALB was significant increased, but the BUN level was decreased in piglet serum by UR administration during pregnancy (P < 0.05).The serum biochemical parameters in sow serum, cord serum and piglet serum were summarized in P < 0.05, P < 0.05, P < 0.05, Sows that were fed the UR-supplemented diet had lower serum contents of Ser and greater serum contents of Cys (P < 0.05). In addition, a significant enhanced in VEGFR2 was also detected in sow placentas (P < 0.05).The expression of amino acids transporters in the placenta of sows was shown in P < 0.05). Importantly, cell viability is the highest at 200, 400, 600, and 1,000 μM of UR.Cell viability was assessed by CCK8 assay. P < 0.05, Since cell cycle progression is also an important regulator of cell proliferation, we next explore whether UR regulated cell cycle . The resMaternal nutrient supply may act as an inducer of fetal development and can permanently affects anatomy, physiology, and metabolism of fetus . Meanwhide novo fatty acid and triglyceride synthesis, vital for the lipogenesis  is related to maternal amino acid metabolism . Nucleotogenesis . It indiogenesis .de novo formation of blood vessels from mesoderm precursor cells, and angiogenesis is creation of new vessels from a preexisting blood supply, which is very important for the maternal-fetal exchange .XW and Y-LY: conceptualization. YL: data curation and writing—original draft. H-LW, LG, and TZ: formal analysis. H-LW and YL: project administration. JL: resources. H-LW, LG, TZ, and XW: writing—review and editing. All authors contributed to the article and approved the submitted version.


Diffuse large B-cell lymphoma (DLBCL) is a highly heterogeneous disease. Therefore, more reliable biomarkers are required to better predict the prognosis of DLBCL. Cuproptosis is a novel identified form of programmed cell death (PCD) that is different from oxidative stress-related cell death  by Tsvetkov and colleagues in a recent study released in Science. Cuproptosis is copper-dependent PCD that is closely tied to mitochondrial metabolism. However, the prognostic value of cuproptosis-related genes (CRGs) in DLBCL remains to be further elucidated. In the present study, we systematically evaluated the molecular changes of CRGs in DLBCL and found them to be associated with prognosis. Subsequently, based on the expression profiles of CRGs, we characterized the heterogeneity of DLBCL by identifying two distinct subtypes using consensus clustering. Two isoforms exhibited different survival, biological functions, chemotherapeutic drug sensitivity, and immune microenvironment. After identifying differentially expressed genes (DEGs) between CRG clusters, we built a prognostic model with the Least absolute shrinkage and selection operator (LASSO) Cox regression analysis and validated its prognostic value by Cox regression analysis, Kaplan-Meier curves, and receiver operating characteristic (ROC) curves. In addition, the risk score can predict clinical characteristics, levels of immune cell infiltration, and prognosis. Furthermore, a nomogram incorporating clinical features and risk score was generated to optimize risk stratification and quantify risk assessment. Compared to the International Prognostic Index (IPI), the nomogram has demonstrated more accuracy in survival prediction. Furthermore, we validated the prognostic gene expression levels through external experiments. In conclusion, cuproptosis-related gene signature can serve as a potential prognostic predictor in DLBCL patients and may provide new insights into cancer therapeutic targets. BaseFDX1, LIAS, LIPT1, DLD, DLAT, PDHA1, and PDHB) that sensitized the cells to cuproptosis by genome-wide CRISPR-Cas9 loss-of-function screen, while three genes  with resistance to cuproptosis. Copper importer (SLC31A1) and copper exporter (ATP7B) have also been found to promote and inhibit cuproptosis, respectively, by regulating intracellular copper concentrations  mentioned above. The differences in survival, drug sensitivity, and immune cell infiltration between subtypes were also integrated. Subsequently, we constructed a prognostic model to stratify patients at risk. Furthermore, we developed a nomogram integrating clinical features and risk scores to quantify risk assessment and predict the overall survival (OS) of DLBCL patients. The results showed that the nomogram was an effective prognostic indicator. Finally, we performed an experimental verification on our clinical samples.22.1https://www.ncbi.nlm.nih.gov/geo/) and The Cancer Genome Atlas (TCGA) . Microarray expression profiles of DLBCL patients were obtained from GSE10846, GSE31312, and GSE87371 datasets using Affymetrix Human Genome U133 Plus 2.0 platform. Transcriptional data for 48 DLBCL samples from TCGA were retrieved from UCSC Xena (https://xenabrowser.net/datapages/). All the microarray data included were normalized and log2 transformed. Probe IDs were mapped to gene symbols according to the corresponding annotation files, expression measurements for all probes associated with the same gene were averaged and the maximum value was finally taken. The pathology image data  slide) were downloaded from the Genomic Data Commons . After excluding samples with missing survival information or survival time of less than one month, the GSE10846, GSE31312, and GSE87371 included 400, 466, and 216 tumor specimens of DLBCL respectively. GSE10846 was used as the training dataset for constructing the subtype and prognostic model, while GSE31312 and TCGA-DLBCL were the validation sets for subtype identification and GSE87371 was the validation for the prognostic model. 12 CRGs  were obtained from the article by Tsvetkov et al.   classification . The stu2.3http://bioinfo.life.hust.edu.cn/GSCA/)  data of these 12 CRGs in all cancers, as well as the copy number variation (CNV) data in DLBCL, were mined in Gene Set Cancer Analysis (GSCA) (n/GSCA/) . We also2.4Consistent unsupervised clustering analysis was performed using the R package “ConsensusClusterPlus”  to class2.5http://www.gsea-msigdb.org/gsea/msigdb). Furthermore, we used the R software package “pRRophetic”  . Time-dependent receiver operating characteristic curve (time-ROC curve) analysis was conducted using the “timeROC” R package  . Then prP <0.01) , 91. Aft package  to obtai2.7The co-expression matrix of CRGs and genes in the prognostic model was established using the “ggcorrplot” package in R. For the DEGs between risk groups identified by the “limma” package, we conducted Gene Ontology (GO) enrichment analysis using the “clusterProfiler” package , 94. Spe2.8A predictive nomogram integrating the clinical characteristics and risk score was developed using the “rms” package according to the outcome of the independent prognosis analysis. Calibration plots of the nomogram were used to measure the consistency of predicted survival events and actual observed results at 1-, 2- and 3 years. Time-ROC curves for 1-, 2- and 3-year survival were performed for the assessment of accuracy in DLBCL prognosis.2.9RNA was extracted from the FFPE samples using the Paraffin-Embedded Tissue RNA Extraction Kit  according to the manufacturer’s instructions. Reverse transcription was performed with the cDNA synthesis kit  to generate cDNAs.2.10Taq Pro Universal SYBR qPCR Master Mix  was then used for quantitative PCR. β-ACTIN was used as an internal control, and each sample was repeated in triplicate. The relative fold-change in expression with respect to a control group was calculated by the 2-ΔΔCt method. The PCR cycle conditions were 95 °C for 30 s, followed by 40 cycles of 95 °C (10 s) and 60 °C (30 s). Three biological replicates were performed. The PCR primers used were as follows:TUBB4A forward primer (FP): 5′‐GAGTTCCCAGACCGCATCA‐3′;TUBB4A reverse primer (RP): 5′‐CGGAAACAGATGTCGTAGAGTG‐3′;SLC38A5 FP: 5′‐AACAGCAATGGAGAGTGAAGC‐3′;SLC38A5 RP: 5′‐ACCTCAGGGTGGCAGACAA‐3′;TEX9 FP: 5′‐TTTATGAGACAGCAGCGAACA‐3′;TEX9 RP: 5′‐GAACCTCTGTGGCACTTTGAC‐3′;S100B FP: 5′‐GGAAGGGAGGGAGACAAGCA‐3′;S100B RP: 5′‐CTGGAAGTCACATTCGCCGT‐3′;ACTIN FP: 5′‐TCAAGATCATTGCTCCTCCTGAG‐3′;ACTIN RP: 5′‐ACATCTGCTGGAAGGTGGACA‐3′;2.11P < 0.05 was considered statistically significant.All statistical analyses were performed with R version 4.1.1, GraphPad Prism 9.0.0, and SPSS software version 26.0. Student’s t-test or one-way analysis of variance was used to analyze differences between groups in variables with a normal distribution. Wilcoxon test was used to analyze the differences between groups of skewed distribution variables. And we used the Kaplan-Meier method for survival analysis and the log-rank test to analyze the differences in overall survival. 33.1CDKN2A was the most frequent mutated CRG (42%), followed by ATP7B, MTF1, GLS, and DLD . Next, we investigated somatic copy number changes in these CRGs of DLBCL and found widespread copy number alterations in all 12 CRGs . Meanwhile, the results showed that the expression of ATP7B, GLS, MTF1, and LIPT1 were negatively correlated with the overall survival (OS) of DLBCL (P<0.05), and the first three genes were previously identified as negative regulators of cuproptosis (P<0.05)  and a possible inhibiting effect on DNA damage (FDR=0.0105) response   (P<0.05)    and B subtype (n=206)  , follicular helper T cells, and activated natural killer (NK) cells were higher in subtype A, while CD8+ T cells, activated CD4+ memory T cells, gamma delta T cells, M1 macrophages, plasma cells had significantly lower infiltration in subtype A compared to those in subtype B. In addition, we also carried out the ssGSEA algorithm in TCGA-DLBCL to further validate the differences in TME cell infiltration between the cuproptosis subtypes .To investigate TME differences in CRGs-related subtypes, the enrichment fraction of 22 kinds of immune cells in both two clusters was evaluated using the CIBERSORT algorithm . SignifiTo better understand the survival differences between the two subtypes, GSVA enrichment analysis of the KEGG pathway was conducted on these two clusters to assess the functional and biological differences. The results showed that the subtype A was mainly enriched in cell signal transduction pathways and immune-related pathways, such as the Notch signaling pathway, MAPK signaling pathway, VEGF signaling pathway, ERBB signaling pathway, primary immunodeficiency, and Fc epsilon RI signaling pathway. B subtype was mainly enriched in the p53 signaling pathway and metabolism-related pathways, including the metabolism of sugar, protein, fat, and nucleotide .We then evaluated the therapeutic responsiveness of chemotherapeutic agents between the two subgroups, subtype A exhibited a resistance tendency to bleomycin, cisplatin, doxorubicin, etoposide, vincristine, vinorelbine and elesclomol, a kind of copper ionophore, while B was more resistant to paclitaxel and methotrexate  was discovered. The optimal weighting coefficient for each gene was determined by the regularization parameter lambda using the min standard . Univariable Cox regression analysis was then performed to acquire 66 DEGs associated with prognosis (p< 0.001). Finally, the candidate genes were incorporated into the construction of the prognostic model using LASSO Cox regression analysis, and a signature of 5 genes (SLC38A5) + (0.0194 × expression of TUBB4A) + (0.0458 × expression of LOC100507477) – (0.0192 × expression of S100B) – (0.0541× expression of TEX9). According to the median risk score of the corresponding datasets, the patients with DLBCL were divided into high-scoring and low-scoring groups respectively in training and validation datasets.The risk model was constructed as follows: risk score = (0.1239 × expression of LOC100507477), and the expression levels of S100B and TEX9 were relatively low, consistent with the above formula    (P<0.05), consistent with the previous results of poor prognosis for subtype A  . A high LIPT1 and PDHB  were positively correlated, while LIPT1 was negatively correlated with TUBB4A . Alluvial diagram is plotted for a better display of clinical characteristics and survival differences between CRG clusters and risk groups , followed by SLC38A5 (29%). The proportion of the CNV for each gene was summarized in To better understand the difference in survival in the prognostic signature, the analysis of the molecular alterations in five genes of the prognostic model was performed. The summary report showed that four of them were mutated at a high frequency in tumor specimens (P <0.01) , lactate dehydrogenase (LDH), extranodal sites (ES), and risk sore  using qRT-PCR. The trend of gene expression in the qRT-PCR analysis was basically consistent with our prognostic model. Compared with the normal lymphoid tissue in the control sample, the expression of risk genes (TUBB4A and SLC38A5) was up-regulated, while the expression of protective genes (S100B and TEX9) was down-regulated in most DLBCL samples (P < 0.05)  and copper chelators  have been applied in anticancer treatment , 98. Cu-n=48), we performed a pan-cancer analysis of the SNVs in 12 CRGs. The results showed that 10 of 12 genes were mutated in 98.23% (941/958) of tumor samples, of which CDKN2A was the most (42%). In addition, CNVs were present in all 12 CRGs. Among them, the most frequent were CDKN2A (predominantly deletion) and FDX1 (predominantly amplification). FDX1 is known to regulate protein lipoylation and is a key regulator in the process of cuproptosis   is a candidate susceptibility gene for lymphoma. Since the products of targets such as metallothionein can suppress cellular stress generated by ionizing radiation (GLS) is a mitochondrial enzyme that catalyzes the conversion of glutamine to glutamate   were assimporter , 110 andimporter . SLC31A1to cells . In addin tumors . Conversreatment , 114. CD2 phases . CDKN2A rognosis –118. Metadiation . Glutamilutamate . Highly lutamate , 122. UnCL cells . These aCDKN2A, MTF1, GLS, and ATP7B) were more highly expressed in subtype A. The difference in the expression of CRGs between the two subtypes may be a potential reason for their distinct prognosis. Next, potential biological differences between subtypes were explored by GSVA. The results showed that subtype A was mainly enriched in pathways closely related to tumor growth and development, such as the Notch signaling pathway, MAPK signaling pathway, VEGF signaling pathway, and ERBB signaling pathway. For instance, Notch2 is a key membrane receptor for B-cell function and plays a critical role in the pathogenesis of lymphoma  using LASSO regression analysis. The model was identified as an independent prognostic factor in Cox regression analysis. Different clinical characteristics and prognoses were demonstrated between high- and low-risk groups. The prognosis of the high-risk group was worse compared to the low-risk group. Subtype A, LDH>ULN, and non-GCB subpopulations tended to have a higher risk score.Based on DEGs between cuproptosis-related subpopulations, we constructed and validated a prognostic model integrating five genes (TUBB4A (45%). All four genes had copy number abnormalities in patients with DLBCL. The human microtubulin β-IVa class (TUBB4A), which pertains to the β-microtubulin family, has little or no expression in most normal tissues but is highly expressed in a variety of human cancer cell lines  and is normally expressed only in the testis, except in early-developing embryos and the placenta. TEX9 expression can be induced in tumor cells when cancer occurs. TEX9 has been shown the promotion of proliferation and migration and has an inhibitory effect on the apoptosis of esophageal squamous carcinoma cells  is a Ca+2/Zn+2-p53 binding protein that blocks phosphorylation and acetylation sites on p53 important for transcriptional activation  of the pan-cancer analysis, with the most variant being ll lines . TUBB4A ignaling . It is aignaling –129. Likma cells . S100 cativation . In additivation . High exrognosis . SLC38A5feration . FurtherAccording to GO enrichment analysis, DEGs between high- and low-risk groups were closely associated with the metal ion transport-related pathways. This suggests that there may be differences in copper transport capacity between the high and low-risk groups, which may be one of the potential reasons for the different prognoses in the two groups. Since serum copper levels have previously been shown to be positively correlated with disease status in non-Hodgkin lymphoma, with significantly higher levels in active or relapsed patients than in patients in remission , 23–25.+ T cells, and activated CD4+ memory T cells. This may have led to the difference in survival between the high- and low-risk groups. Ultimately, a nomogram integrating clinical features and risk scores was constructed. IPI is the main clinical tool used to predict the prognosis of patients with aggressive NHL and is the current prognostic benchmark for DLBCL . qRT-PCR analysis showed that the expression of risk genes (TUBB4A and SLC38A5) was up-regulated, while the expression of protective genes (S100B and TEX9) was down-regulated in DLBCL. This was generally consistent with our prognostic model.To further verify the expression level of signature genes in DLBCL, we performed qRT-PCR on our clinical samples to quantify the mRNA expression of these four genes (Our study has several limitations. Firstly, our clustering typing strategy and prognostic signature need to be further validated for their robustness and clinical utility in a larger sample. Secondly, since the establishment and validation of the prognostic model were primarily based on public databases, further validations are required through cell experiments and larger clinical samples. In addition, the specific mechanism of CRG in DLBCL and the underlying mechanism between CRGs and tumor immunity in DLBCL are currently unclear and require further study.In conclusion, we performed systematic analyses on the molecular alterations of CRGs in DLBCL, and our study suggests that these genes may play a key role in the prognosis of DLBCL. The two subtypes identified based on the CRGs expression signature were significantly different in biological function, immune cell infiltration, treatment responsiveness, and clinical prognosis. In addition, the prognostic model constructed from CRG performed well in predicting the survival of DLBCL patients and was significantly correlated with the level of immune infiltration. Furthermore, we built a nomogram combining clinical features and risk scores that improved the predictive power of DLBCL. Finally, we carried out an external experiment to verify the level of prognostic gene expression. Our work provides new directions for prognostic prediction and potential therapeutic targets in DLBCL and may provide the basis for more in-depth studies in the future.The original contributions presented in the study are included in the article/BZ conceived the study design, completed data analysis and interpretation, and manuscript writing. QW assisted in the completion of experiments. TZ, ZZ, ZL, DZ, and ZC provided suggestions for the data analysis. YM provided critical revisions to the paper. All authors contributed to the article and approved the submitted version.


Previous research on the psychological mechanisms of obesity has primarily focused on acute psychopathology. However, there is limited literature on the role of more complex and entrenched psychological processes in weight management. The current study aimed to expand previous research by examining more enduring psychological constructs, including early maladaptive schemas (EMS), schemas modes, and trauma.N = 125) comprised adults with normal weight (n = 40) and obesity (n = 85) from community and clinical settings in Australia. Eligible participants completed a series of self-report questionnaires via Research Electronic Data Capture (REDCap). Two, separate, one-way multivariate analysis of variance (MANOVA) were conducted to examine group differences on the outcome variables.Participants  = 2.97, p < .001, partial η2 = .51. Follow-up univariate tests revealed that individuals with obesity endorsed significantly more maladaptive schemas and schema modes and significantly less healthy schema modes than individuals with normal weight. In addition, results demonstrated a significant effect of group on childhood trauma and posttraumatic stress disorder (PTSD) symptoms, V = .19, F = 4.70, p < .001, partial η2 = .19. Subsequent univariate tests and chi-square analyses indicated that individuals with obesity reported significantly more childhood trauma as well as significantly more PTSD symptoms within the last month than normal weight individuals.Findings indicated a significant effect of group on EMS and schema modes, V = .51, This was the first study to compare EMS and schema modes in treatment-seeking individuals with obesity and normal weight controls using the short form version 3 of the Young Schema Questionnaire and revised, 118-item, Schema Mode Inventory. Overall, findings revealed that individuals with obesity experience more complex and enduring psychological difficulties than normal weight individuals. Increased assessment and targeted treatment of these underlying mental health concerns may contribute to a more holistic conceptualisation of obesity and could improve the long-term success of weight management.The online version contains supplementary material available at 10.1186/s12888-022-04169-7. Preventing weight regain after weight loss intervention remains the single greatest challenge for individuals with obesity –3. SucceEMS are central to schema therapy, an integrated psychotherapy incorporating and extending elements of cognitive-behavioural therapy, Gestalt therapy, psychodynamic therapy, and attachment theory . EMS refAnother key concept in schema therapy is the notion of schema modes. Schema modes are moment-to-moment coping responses and emotional states associated with one or several EMS  to compare EMS in individuals with overweight and obesity and normal weight controls –17. FirsTo date, no study has utilised the Young Schema Questionnaire, Short Form, Version 3 (YSQ-S3) to compare EMS in individuals with obesity and normal weight controls. In addition, previous studies using the YSQ-S2 have found inconsistent results regarding group differences. Further, the only study examining schema modes in obesity combined overweight and obesity participants from the community and utilised an earlier iteration of the Schema Mode Inventory, containing 124-items . To our The goal of the current study was to examine whether there were significant differences in EMS and schema modes among treatment-seeking individuals with obesity and normal weight controls. This study extends previous research by comparing a normal weight control group to individuals with obesity attending a tertiary weight intervention service, and by utilising the latest iteration of the schema questionnaires, namely the YSQ-S3 and revised SMI. In addition, we aimed to investigate whether individuals with obesity and normal weight significantly differed on childhood trauma and PTSD symptoms. In light of previous research, we hypothesised that the obesity group would have significantly higher scores on the Abandonment, Defectiveness/Shame, Social Isolation/Alienation, Dependence/Incompetence, Failure, Insufficient Self-Control, and Subjugation schemas relative to normal weight controls. Given the existing literature on trauma and obesity, we also hypothesised that individuals with obesity would have significantly higher scores on the Mistrust/Abuse and Emotional Deprivation schemas compared to normal weight individuals. In addition, we predicted that individuals with obesity would have significantly higher scores on the Vulnerable Child, Impulsive Child, and Detached Protector schema modes, but significantly lower scores on the Happy Child and Healthy Adult modes compared to normal weight controls. Further, we hypothesised that the obesity group would have significantly higher scores on childhood trauma and PTSD symptoms compared to the normal weight group.N = 163) comprised adults with normal weight (n = 58) and obesity (n = 105) from Australia. This data overlaps with another study submitted for publication by the same authors, though focuses on distinct research questions and statistical analyses. Normal weight participants were recruited via community advertisements and social media. Participants with normal weight were eligible if aged 18 years or older and BMI was between 18.5 to 24.9 kg/m2. Participants with obesity were recruited from the Blacktown Metabolic and Weight Loss Program (BMWLP) at Blacktown Hospital, prior to commencing treatment. The BMWLP is a tertiary level service that provides multidisciplinary weight intervention for adults with obesity, including the option for bariatric surgery for eligible patients. To be eligible for enrolment in the BMWLP, participants required a BMI 2 with type 2 diabetes or a BMI 2 with at least two obesity-related complications . Recruitment occurred across a 12-month period. All participants were informed about the research aims. Participants in either group were excluded if they had a cognitive or speech impairment, neurological disorder, intellectual disability, head injury, or mental health condition which interfered with their ability to complete or understand the requirements of the study. Eligible participants completed a series of self-report questionnaires via Research Electronic Data Capture (REDCap), a secure web-based software platform [Participants (platform . The queN = 125) included males (32.8%) and females (67.2%) that ranged in age from 21 to 79 years  and ranged in BMI from 19.1 kg/m2 to 75.4 kg/m2 . Of these 125 participants, 40 (32%) were in the normal weight group and 85 (68%) were in the obesity group. Table Of the 163 participants, 37 were excluded because of missing data. One significant outlier was also excluded. The final sample  was collected through a general self-report questionnaire. Mental and physical health information was also collected to determine participant eligibility.2).Specialised wheelchair scales were used to measure participant height and weight for the obesity group. Self-reported height and weight was used for the normal weight group due to changes in recruitment procedures associated with COVID-19. BMI was computed by dividing weight in kilograms by height in metres squared (kg/mcompletely untrue of me) to 6 (describes me perfectly). A total score for each scale is derived by summing the scale items. Higher scores indicate greater endorsement of that schema. The YSQ-S3 has demonstrated good internal consistency in clinical and non-clinical samples across several languages ..completenever or almost never) to 6 . The score for each scale is derived by calculating the scale mean. Higher scores indicate greater endorsement of that schema mode. The SMI has demonstrated acceptable internal consistency and test–retest reliability [The SMI is a 118-item self-report measure of moment-to-moment coping responses and cognitive and emotional states . The SMIiability .never true) to 5 (very often true). The total score for each scale ranges from 5 to 25 with higher scores suggesting greater exposure to maltreatment. Two main approaches have been applied to interpreting the CTQ-SF. First, the total score for each scale can be classified according to four levels of maltreatment: None , Low (to Moderate), Moderate (to Severe), and Severe (to Extreme) [very often true) were coded as 1 (possible underreporting), while items rated 1 (never true) through 4 (often true) were coded as 0 (no underreporting). The validity and reliability of the CTQ-SF has been well-supported across both clinical and non-referred samples [The CTQ-SF is a 28-item self-report screening measure of retrospective childhood abuse and neglect . The CTQExtreme) . Second,Extreme) . Using treatment . The CTQ samples .not at all) to 4 (extremely). A total score ranging from 0 to 80 is calculated by summing the items. Higher scores indicate a greater frequency of PTSD symptoms. A provisional PTSD diagnosis is indicated using a cut-off score of 31 to 33. In the current study, participants scoring above 31 were coded as 1 , while participants scoring below 31 were coded as 0 (no diagnosis). Each item also corresponds with a symptom of PTSD in the Diagnostic and Statistical Manual of Mental Disorders-fifth edition (DSM-5) [moderately) or higher indicate a DSM-5 symptom has been endorsed. The PCL-5 has demonstrated high internal consistency, test–retest reliability, and convergent and discriminant validity [The PCL-5 is a 20-item self-report measure of PTSD symptoms within the past month . Items a (DSM-5) . Correspvalidity , 31.2) analyses were performed to investigate whether groups differed on childhood trauma and provisional PTSD diagnosis using the dichotomous scoring method on the CTQ-SF and PCL-5, respectively. Pillai’s trace (V) was used to interpret the MANOVA results. Pillai’s trace has been recommended as the statistic of choice because of its robustness to model violations [2) was used as the effect size . Bonferroni correction was applied to circumvent inflation of the type 1 error rate due to multiple comparisons. The alpha level for the first and second MANOVA were adjusted to .002 and .008, respectively.All analyses were carried out using SPSS Version 28.0. Initially, the data was inspected for outliers and missing values, and assumption testing was performed. Missing data was managed through listwise deletion. Two, separate, one-way multivariate analysis of variance (MANOVA) were conducted to examine group differences on the outcome variables. The choice to conduct two separate MANOVA was based on the theoretical and empirical relationship between these variables, and the research aims. It was also anticipated that two separate MANOVA would likely yield the most clinically meaningful interpretations given the paucity of research in this area. Further, fitting separate models has been recommended when there are reasonable theoretical grounds for separating outcome measures . First, olations –35. FollN = 125) was utilised to examine group differences on the outcome variables. All observations were independent, measured on a ratio scale, and randomly sampled from the populations of interest. There were 33 (38.8%) participants in the obesity group and 10 (25%) participants in the normal weight group with scores on the CTQ-SF that suggested possible underreporting of maltreatment . These participants were retained in the analyses. There were no concerns with multicollinearity and there was sufficient observed power (> .80) for all significant effects. Multivariate normality and homogeneity of covariance matrices were partially satisfactory. These model assumptions, however, are susceptible to deviations in large samples [Data from the total sample  = 2.97, p < .001, partial η2 = .51. As hypothesised, separate univariate tests on the outcome variables revealed that individuals with obesity had significantly higher scores on the Abandonment (p < .001), Mistrust/Abuse (p < .001), Emotional Deprivation (p < .001), Defectiveness/Shame (p < .001), Social Isolation/Alienation (p < .001), Dependence/Incompetence (p < .001), Failure (p < .001), Insufficient Self-Control (p < .001), and Subjugation (p < .001) schemas compared to normal weight controls. Unexpectedly, individuals with obesity also had significantly higher scores on the Vulnerability to Harm and Illness (p < .001), Self-Sacrifice (p = .002), Negativity/Pessimism (p < .001), and Emotional Inhibition (p < .001) schemas. Furthermore, as predicted, individuals with obesity demonstrated significantly higher scores on the Vulnerable Child (p < .001), Impulsive Child (p < .001), and Detached Protector (p < .001) schema modes as well as significantly lower scores on the Happy Child (p < .001) and Healthy Adult (p < .001) modes compared to normal weight controls. Unexpectedly, individuals with obesity also exhibited significantly higher scores on the Angry Child (p = .002), Undisciplined Child (p < .001), Compliant Surrender (p < .001), and Punitive Parent (p < .001) modes. Table Using Pillai’s trace, there was a significant effect of group on EMS and schema modes, V = .51, F3, 92 = 2.F = 4.70, p < .001, partial η2 = .19. As hypothesised, separate univariate tests on the outcome variables revealed that individuals with obesity had significantly higher scores on Emotional Abuse (p < .001), Physical Abuse (p < .001), Sexual Abuse (p = .005), Emotional Neglect (p = .004), Physical Neglect (p < .001), and PTSD symptoms (p < .001) compared to normal weight controls. Table Using Pillai’s trace, there was a significant effect of group on childhood trauma and PTSD symptoms, V = .19, 2  = 6.18, p = .013, Physical Abuse, χ2  = 8.16, p = .004, Sexual Abuse, χ2  = 8.91, p = .003, Emotional Neglect, χ2  = 8.78, p = .003, Physical Neglect χ2  = 14.93, p < .001, and provisional PTSD diagnosis, χ2  = 6.59, p = .010, compared to normal weight individuals. Table Using the dichotomous scoring method on the CTQ-SF and PCL-5, chi-square analyses revealed that individuals with obesity reported a significantly higher proportion of Emotional Abuse, χThe current study was the first to compare early maladaptive schemas (EMS) and schema modes using the YSQ-S3 and revised SMI, respectively, in normal weight controls and individuals with obesity attending a tertiary weight intervention service. This study also investigated differences in childhood trauma and PTSD symptoms among treatment-seeking individuals with obesity and normal weight controls.First, consistent with our hypotheses and previous research , 16, indSpecifically, these results suggested that compared to normal weight individuals, those with obesity may perceive their important relationships with others as more unstable  or may believe that others are more likely to abuse and treat them poorly  and that their emotional needs will remain unsatisfied  , 8. TheySecond, consistent with our predictions and previous research , individThird, as hypothesised, individuals with obesity demonstrated significantly higher scores on Emotional Abuse, Physical Abuse, Sexual Abuse, Emotional Neglect, Physical Neglect, and PTSD symptoms compared to normal weight individuals. Significant differences between groups were also observed on childhood trauma and provisional PTSD diagnosis using the dichotomous scoring methods on the CTQ-SF and PCL-5. Specifically, 58.8% of individuals with obesity reported the presence of Emotional Abuse, 49.4% reported the presence of Physical Abuse, 28.2% reported the presence of Sexual Abuse, 31.8% reported the presence of Emotional Neglect, 54.1% reported the presence of Physical Neglect, and 37.6% reported PTSD symptoms within the last month that were above the cut-off criteria to suggest a provisional diagnosis of PTSD. These results support previous studies that found an increased probability of developing obesity for those with childhood traumatic experiences and PTSD –12, 38. Previous research on the psychological mechanisms of obesity has primarily concentrated on acute psychopathology. This study contributes to elucidating the relationship between obesity and more complex and entrenched psychological processes. Overall, our findings indicated that individuals with obesity endorsed significantly more maladaptive schemas and schema modes and significantly less healthy schema modes than individuals with normal weight. In addition, individuals with obesity reported significantly more childhood trauma as well as significantly more PTSD symptoms within the last month than normal weight individuals. These findings have several clinical implications for the management of obesity and mental health more broadly.First, they highlight the importance of comprehensively assessing maladaptive schemas and schema modes in weight intervention programs. Identifying how patients view themselves, others, and the world, and their primary coping strategies, may provide valuable insight for targeted intervention. For instance, our findings revealed that individuals with obesity principally display a psychologically withdrawn and submissive or passive coping style. This suggests that patients have developed ways to emotionally detach from their experiences, but do not employ effective strategies to manage their emotional difficulties. These maladaptive coping mechanisms may be activated by underlying schemas and schema modes that are entrenched within the patient . This formulation is conceptually consistent with the schema mode model of obesity proposed by Basile et al. , which tSecond, as part of weight intervention, maladaptive schemas and schema modes should be addressed through psychological treatments such as schema therapy. Schema therapy integrates cognitive, behavioural, and experiential techniques to modify maladaptive coping strategies, reduce the influence of the internalised parent modes, satisfy unmet core emotional needs, and foster healthy functioning , 8. ImpoThird, these findings emphasise the importance of comprehensively assessing trauma history in weight intervention programs. Research indicates that individuals with obesity with a history of childhood abuse and PTSD demonstrate significantly more eating psychopathology, physical health concerns, and psychological difficulties  than those without a history of trauma or PTSD . FurtherFinally, these findings reinforce the importance of treating mental health difficulties in weight intervention programs. Psychological difficulties  have been linked to weight regain after bariatric surgery , 42 and Our results should be considered in the context of several limitations. First, we did not explicitly assess all types of childhood maltreatment . As a result, some participants may have experienced childhood adversities that were not captured in this study. For example, MacDonald et al.  found thSecond, responses on schema-related questionnaires may be influenced by active schemas or schema modes at the time of the assessment. For example, individuals in an avoidant coping mode  may inadvertently minimise their clinical symptoms, whereas individuals in a Vulnerable Child mode may unintentionally inflate their concerns. Similarly, although EMS are considered relatively stable, it is unclear whether certain questions  prime responses or elicit emotional states that influence responding style. In addition, it is unclear whether certain psychiatric medication attenuates schema activation. The potential variability in schema activation at the time of the assessment may contribute to differences in outcomes across the literature, but future research is required to verify this claim. In addition, future research could examine these outcomes over multiple time points to circumvent potential response bias associated with active schemas and schemas modes.Third, previous research has shown that treatment-seeking individuals with obesity may underreport their clinical symptoms due to fear of treatment ineligibility . SupportFourth, our sample predominantly consisted of female participants, with a large portion recruited from a clinical setting. Notably, this clinical setting resides in a lower socioeconomic area in Australia than the sample recruited by Anderson et al.  and is cFinally, there are several notable strengths in the current study. First, we utilised the YSQ-S3 and refined, 118-item SMI, which are the latest iterations of the schema questionnaires. This extends previous literature that compared EMS and schema modes in individuals with obesity and normal weight controls using the YSQ-S2 and 124-item Schema Mode Inventory, respectively –17. ImpoThis was the first study to utilise the YSQ-S3 and revised, 118-item, SMI to compare EMS and schema modes in normal weight controls and individuals with obesity attending a tertiary weight intervention service. This study also examined differences in childhood trauma and PTSD symptoms among treatment-seeking individuals with obesity and normal weight controls. Overall, results revealed that individuals with obesity endorsed significantly more maladaptive schemas and schema modes and significantly less healthy schema modes than individuals with normal weight. In addition, individuals with obesity reported significantly more childhood trauma as well as significantly more PTSD symptoms within the last month than normal weight individuals. Future obesity treatment models should incorporate routine assessment of these psychological difficulties, which may contribute to a more holistic conceptualisation of obesity. Moreover, addressing these underlying mental health concerns through evidence-based psychological interventions such as schema therapy, may attenuate the influence of psychological processes on weight management, and could improve the long-term success of patients with obesity.Additional file 1: Additional file 2:


We performed a prospective study on peritoneal dialysis (PD) to investigate the relationship between PCS or IS levels with clinical outcomes.The impact of This prospective cohort study investigated the association of serum PCS and IS with clinical outcomes in patients undertaking PD. We performed a correlations analysis to explore the influencing factors of PCS an IS. Meta-analysis was conducted to objectively evaluate the prognostic effects of PCS and IS on different stages of CKD patients.p < 0.001), but also increased the risk of cardiovascular event  and PD-associated peritonitis . Dividing the total PCS level by 18.99 mg/L, which was calculated from the best cutoff value of the ROC curve, patients with total PCS higher than 18.99 mg/L had worse prognosis. Meta-analysis confirmed its value in cardiovascular event in PD.A total of 127 patients were enrolled consecutively and followed with an average period of 51.3 months. Multivariate Cox regression showed that serum total PCS not only contributed to the occurrence of PD failure event (HR: 1.05, 95% CI = 1.02 to 1.07, The serum total PCS concentration was a detrimental factor for higher PD failure event, cardiovascular event, and PD-associated peritonitis. It could be used as an innovative marker in predicting poor clinical outcome in PD. Impairment of renal function results in the retention of large numbers of compounds, as known as uremic toxins or uremic retention solutes, which normally can be excreted to the urine . In 2012p-cresyl sulfate (PCS) and Indoxyl sulfate (IS) are two major components [Among the various members of PBUTs, mponents ,8. Both mponents . Colonicmponents ,4.In recent years, PBUTs have gained more and more attention as a potential hazard leading to worse prognosis, however, the prognostic effect of PCS or IS on different stages of CKD patients still remains debatable. There were reports about high PCS levels that increased the risk of cardiovascular diseases in pre-dialysis CKD ,10 and awww.miraclamedicle.com:1613). We recruited 127 patients with PD starting from January 2012. Patients aged > 18 but < 75 years old undertaking continuous ambulatory peritoneal dialysis (CAPD) or automatic peritoneal dialysis (APD) longer than 3 months were eligible for inclusion. Patients were excluded when they were having (a) peritonitis in the past 1 month; (b) acute infection and cardiovascular event in the past 3 months; (c) active stage of autoimmune diseases; (d) having suffering malignancy. The study was performed in accordance with the principles of the Declaration of Helsinki and approved by the Ethics Committee of Changzheng Hospital (ChiCTR-IOR-14005541).The prospective study was conducted using the PD Information System of Changzheng Hospital, Shanghai , Albumin (g/L), Creatinine (µmol/L), Urea nitrogen (mmol/L), Uric acid (µmol/L), Calcium (mmol/L), Phosphate (mmol/L), Bicarbonate (mmol/L), and PTH (pg/mL). Kt/V and creatinine clearance rate (Ccr) for PD were calculated at the time of the first peritoneal equilibration test after signing consent for all patients. All serum IS and PCS were measured two times to obtain an average value. The first measurement of serum PCS and IS was performed at the beginning of recruitment, and the second was performed at the third month after the first. The metabolic syndrome (MS) is defined consistently as previously published literature .r > 0.99) and 50 to 10,000 ng/mL for PCS (r > 0.99). The limit of detection was 500 ng/mL for IS and 50 ng/mL for PCS. Relative standard deviation (SD) of intra- and inter-day precision was with ± 15%.The serum concentrations of total and free PCS and IS were determined by HPLC/MS/MS method as previously described . Serum s3 with neutrophils leukocytes 50% or more on a 2-h dwell sample; and (iii) positive effluent Gram staining [Patients were followed-up and visited the PD clinic at 3–6-month intervals with clinical and biochemical data recorded. Clinical endpoints were prospectively collected. The patient was assumed to be lost to follow-up starting from the date of the last actual visit. The major endpoint was PD failure event, which was defined as a composite of switching to HD or all-cause mortality during the study period. The secondary endpoints were cardiovascular event and time-to-first PD-associated peritonitis. Cardiovascular event was defined as patients with any one of the following including death from cardiac causes, myocardial ischemia, nonfatal myocardial infarction, ischemia stroke, or new onset of peripheral vascular disease, whichever occurred first ,9. PD-asstaining . The earMeta-analysis was performed to objectively evaluate the prognostic impact of PCS and IS on different stages of CKD, including pre-dialysis and dialysis patients. We searched the PubMed, EMBASE, Cochrane, and Web of Science databases with terms including ‘uremic toxins’, ‘protein-bound toxins’ or ‘PCS’ combined with the terms ‘chronic kidney disease’ or ‘CKD’ or ‘end stage renal disease’ or ‘ESRD’ or ‘dialysis’. The systematic literature review was screened with data extracted meanwhile independently by Zewei Chen and Jing Xu.t tests. Spearman’s rank correlation or Pearson’s correlation coefficient were used to estimate the relationships between serum PCS or IS levels and selected variables. Cox regression model was used to analyze the relationship between independent variables and clinical outcomes, and reported by Hazard ratio (HRs) with 95% confidence intervals (CIs). The ROC curve was used to estimate the best cutoff serum concentrations of total-PCS of PD failure event. The Kaplan-Meier method (factors were compared using the log-rank test) was used to estimate cumulative survival rate of time to clinical endpoints in PD patients with total PCS above and below the best cutoff point. For the meta-analysis, adjusted point estimates from each study were consolidated by the generic inverse variance approach, which designated the weight of each study based on its variance. Forest plots for outcomes were generated with 95% CIs. Given the possibility of between-study variance, we adopted a random-effects model rather than a fixed-effect model. Date was analyzed using the SPSS 26.0 software and Review Manager 5.3. The authors had full access to the data and take responsibility for its integrity. p < 0.05 was considered statistically significant.Continuous variables were expressed as mean ± SD for normally distributed variables or median (interquartile range) otherwise. Categorical variables are expressed as frequencies (percentage). Comparisons of continuous and categorical data for patients in different groups were performed by independent samples A total of 127 patients undertaking sustained PD were recruited consecutively from January 2012 to February 2015 and ended on February, 2020 with an average age of 44.7 ± 13.9 years old, and included 77 males (60.6%). The etiology of ESRDs was chronic glomerulonephritis, nephrotic syndrome, IgA nephropathy, diabetic nephropathy, hypertensive nephropathy, or polycystic kidney disease. The demographic and clinical characteristics were shown in p < 0.001, p < 0.001, p = 0.002, respectively) than those without.During the study period , 34 (26.8%) patients experienced PD failure event, among whom 26 transferred to HD due to inadequate dialysis or recurrent peritonitis. Eight patients (6.3%) died during follow-up. In addition, 17 (13.4%) patients experienced PD-associated peritonitis, and 10 (7.9%) had newly occurred cardiovascular event. The distribution of total PCS and IS levels by different endpoints was shown in p < 0.001), age , and peritoneal Kt/V  were independently associated with PD failure event, as showed in p = 0.001) and higher total PCS  increased the risk of having cardiovascular event. As for PD-association peritonitis, total PCS  was the only independent factor. The serum total and free IS was nonrelated to any clinical outcome in univariate and multivariate analyses. These results showed that total PCS played a critical role in clinical outcomes of PD patients.Multivariate cox regression analysis revealed that total PCS . Moreover, patients with high total PCS levels had higher risk of cardiovascular event (log rank p = 0.001) and PD-association peritonitis (log rank p = 0.001) as well during 5-year follow-up.A best cutoff point to evaluate the accuracy of predicting 5-year PD failure event from serum total PCS were obtained by ROC analysis . The arer = −0.397, p = <0.001) and Ccr , while not for PCS. Meanwhile, total IS levels were positively co-related to peritoneal Kt/V  and Ccr , whereas not for PCS either. Other variables associated with total IS levels were diabetes mellitus, albumin, and creatinine.Total and free PCS and IS levels were interrelated . Total Ip = 0.002, p < 0.001, p < 0.001, p < 0.001, A total of 11 prospective cohort studies were included and the baseline characteristics were shown in The clinical significance of PBUTs in PD patients needs further investigation. Our study showed that serum total PCS, one of major types of PBUTs, not only contributed to the occurrence of PD failure event, but also increased the risk of cardiovascular event; whereas, it did not contributed to higher all-cause mortality in PD. These observations were similar to the findings reported by Lin et al. . In addiWe did the further meta-analysis to explore the influence of PCS and IS on prognosis of different stages of CKD, including pre-dialysis CKD, HD, and PD patients. The overall effect of elevated serum total PCS was significantly associated with higher risk of cardiovascular event and all-cause mortality including dialysis and nondialysis subjects. Intriguingly, subgroups analysis in patients undertaking PD revealed that total PCS was responsible for the cardiovascular event, but not for all-cause mortality , which wp-cresol or indoxyl can be increased or decreased by dietary protein, carbohydrates, and fiber [Considering the diverse effect of serum PCS and IS on patients’ prognosis in PD, we analyzed the potent influential factors of the two major types of PBUTs . We obsend fiber ,23,24; hnd fiber ,25. FurtIn recent years, therapeutic strategies have also been developed aiming at lowering PBUTs, mainly represented by the serum concentrations of PCS and IS in CKD patients to observe their beneficial effects, including prebiotic , CurcumiThe major limitation of the study presented here is that all subjects in our study were enrolled from one medical center and sample bias existed due to its nature. The statistical power was limited because of the small sample size. Moreover, some of the concomitant medications changed frequently in the PD patients during the follow-up when they came to the out-patient department; thus we did not include them as parameters in the prognosis analysis; however, they might be the influential factors for serum PCS and IS levels, which we will pay more attention to in our future study design.The serum total PCS concentration was a detrimental factor for higher dialysis failure event, cardiovascular event, and PD-associated peritonitis in sustained PD patients. It could be used as an innovative marker in predicting poor clinical outcomes in PD. Further studies are necessary to investigate the relationship between therapeutic methods of lowering the serum PCS levels and improving clinical outcomes of pre-dialysis CKD and dialysis patients.