Identification of Comprehensive Biomarkers in Patients With Mismatch Repair-Deficient Colon Adenocarcinoma Based on Parallel Multiomics.

Immunocheckpoint inhibitors have shown impressive efficacy in patients with colon cancer and other types of solid tumor that are mismatch repair-deficient (dMMR). Currently, PCR-capillary electrophoresis is one of the mainstream detection methods for dMMR, but its accuracy is still limited by germline mismatch repair (MMR) mutations, the functional redundancy of the MMR system, and abnormal methylation of MutL Homolog 1 promoter. Therefore, this study aimed to develop new biomarkers for dMMR based on artificial intelligence (AI) and pathologic images, which may help to improve the detection accuracy. To screen for the differential expression genes (DEGs) in dMMR patients and validate their diagnostic and prognostic efficiency, we used the expression profile data from the Cancer Genome Atlas (TCGA). The results showed that the expression of Immunoglobulin Lambda Joining 3 in dMMR patients was significantly downregulated and negatively correlated with the prognosis. Meanwhile, our diagnostic models based on pathologic image features showed good performance with area under the curves (AUCs) of 0.73, 0.86, and 0.81 in the training, test, and external validation sets (Jiangsu Traditional Chinese Medicine Hospital cohort). Based on gene expression and pathologic characteristics, we developed an effective prognosis model for dMMR patients through multiple Cox regression analysis (with AUC values of 0.88, 0.89, and 0.88 at 1-, 3-, and 5-year intervals, respectively). In conclusion, our results showed that Immunoglobulin Lambda Joining 3 and nucleus shape-related parameters (such as nuclear texture, nuclear eccentricity, nuclear size, and nuclear pixel intensity) were independent diagnostic and prognostic factors, suggesting that they could be used as new biomarkers for dMMR patients.

GE-Impute: graph embedding-based imputation for single-cell RNA-seq data.

Single-cell RNA-sequencing (scRNA-seq) has been widely used to depict gene expression profiles at the single-cell resolution. However, its relatively high dropout rate often results in artificial zero expressions of genes and therefore compromised reliability of results. To overcome such unwanted sparsity of scRNA-seq data, several imputation algorithms have been developed to recover the single-cell expression profiles. Here, we propose a novel approach, GE-Impute, to impute the dropout zeros in scRNA-seq data with graph embedding-based neural network model. GE-Impute learns the neural graph representation for each cell and reconstructs the cell-cell similarity network accordingly, which enables better imputation of dropout zeros based on the more accurately allocated neighbors in the similarity network. Gene expression correlation analysis between true expression data and simulated dropout data suggests significantly better performance of GE-Impute on recovering dropout zeros for both droplet- and plated-based scRNA-seq data. GE-Impute also outperforms other imputation methods in identifying differentially expressed genes and improving the unsupervised clustering on datasets from various scRNA-seq techniques. Moreover, GE-Impute enhances the identification of marker genes, facilitating the cell type assignment of clusters. In trajectory analysis, GE-Impute improves time-course scRNA-seq data analysis and reconstructing differentiation trajectory. The above results together demonstrate that GE-Impute could be a useful method to recover the single-cell expression profiles, thus enabling better biological interpretation of scRNA-seq data. GE-Impute is implemented in Python and is freely available at https://github.com/wxbCaterpillar/GE-Impute.

Helicobacter pylori induces urease subunit B-specific CD8+ T cell responses in infected individuals via cytosolic pathway of cross-presentation.

BACKGROUND: Urease subunit B (UreB), a conserved and key virulence factor of Helicobacter pylori (H. pylori), can induce the host CD4+ T cell immune responses to provide protection, but less is known regarding CD8+ T cell responses. The characteristics of H. pylori-specific CD8+ T cell responses and the mechanism underlying antigen processing and presentation pathways remain unclear. This study was focus on protective antigen recombinant UreB (rUreb) to detect specific CD8+ T cell responses in vitro and elucidate the mechanism of UreB antigen processing and presentation. METHODS: The peripheral blood mononuclear cells (PBMCs) collected from H. pylori-infected individuals were stimulated with rUreB in vitro to detect specific CD8+ T cell responses after co-culture with rUreB-pulsed autologous hMDCs. Through blocking assay, we investigated the potential pathway of UreB antigen processing and presentation via the cytosolic pathway or vacuolar pathway. The cytokines production of UreB specific CD8+ T cell were evaluated as well. RESULTS: We demonstrated UreB can induce specific CD8+ T cell immune responses in H. pylori infected individuals. Importantly, we characterized that UreB were mainly processed by proteasome instead of lysosomal proteases and presented through cytosolic pathway of cross-presentation, which requires endoplasmic reticulum-Golgi transport and newly synthesized MHC-I molecules, to induce functional-specific CD8+ T cell (IFN-γ + TNF-α + Grz A+ Grz B+) responses. CONCLUSIONS: These results suggest that H. pylori UreB induces specific CD8+ T cell responses through cytosolic pathway of cross-presentation in infected individuals.

Development and validation of a nomogram for postoperative sleep disturbance in adults: a prospective survey of 640 patients undergoing spinal surgery.

BACKGROUND: Postoperative sleep disturbance (PSD) is a prevalent clinical complication that may arise due to various factors. The purpose of this investigation is to identify the risk factors for PSD in spinal surgery and establish a risk prediction nomogram. METHODS: The clinical records of individuals who underwent spinal surgery from January 2020 to January 2021 were gathered prospectively. The least absolute shrinkage and selection operator (LASSO) regression, along with multivariate logistic regression analysis, was employed to establish independent risk factors. A nomogram prediction model was devised based on these factors. The nomogram's effectiveness was evaluated and verified via the receiver operating characteristic (ROC) curve, calibration plot, and decision curve analysis (DCA). RESULTS: A total of 640 patients who underwent spinal surgery were analyzed in this investigation, among which 393 patients experienced PSD with an incidence rate of 61.4%. After conducting LASSO regression and logistic regression analyses using R software on the variables in training set, 8 independent risk factors associated to PSD were identified, including female, preoperative sleep disorder, high preoperative anxiety score, high intraoperative bleeding volume, high postoperative pain score, dissatisfaction with ward sleep environment, non-use of dexmedetomidine and non-use of erector spinae plane block (ESPB). The nomogram and online dynamic nomogram were constructed after incorporating these variables. In the training and validation sets, the area under the curve (AUC) in the receiver operating characteristic (ROC) curves were 0.806 (0.768-0.844) and 0.755 (0.667-0.844), respectively. The calibration plots indicated that the mean absolute error (MAE) values in both sets were respectively 1.2% and 1.7%. The decision curve analysis demonstrated the model had a substantial net benefit within the range of threshold probabilities between 20% and 90%. CONCLUSIONS: The nomogram model proposed in this study included eight frequently observed clinical factors and exhibited favorable accuracy and calibration. TRIAL REGISTRATION: The study was retrospectively registered with the Chinese Clinical Trial Registry (ChiCTR2200061257, 18/06/2022).

Excimer laser model based on a temporal convolutional neural network.

Excimer lasers play a key role in deep ultraviolet lithography. To further study the voltage and energy features of the excimer laser and control it to work in a constant energy mode, a temporal convolutional neural network was designed to fabricate an excimer laser voltage-energy model. The proposed model uses the currently measured energy data to predict the subsequent output energy data. For the voltage-energy data that cannot be obtained, we simulated the initial energy data corresponding to the part of the voltage value based on the relationship between energy and voltage as the initial input of the model. The energy data of any voltage for the excimer laser at each moment were obtained. Finally, a continuous excimer laser voltage-energy model was established. The difference between the means of the measured and generated energy data is less than 0.5 mJ.

Analytical performance evaluation of different test systems on serum creatinine assay.

BACKGROUND: Serum creatinine (SCr) is a useful diagnostic marker for the assessment of renal function. Accurate quantitation of SCr is clinically important in calculation of glomerular filtration rate (GFR). METHOD: To confirm whether there are differences in SCr between enzymatic kits of different manufacturers, the analytical performance of the matched and open test system in the measurement of SCr was evaluated. The analytical performance evaluation was conducted according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Precision, accuracy, linearity, dilution, lower limit of measurement and analytical interference were studied between the two test systems. RESULTS: The performance of SCr from the open test system was in compliance with the matched test system with good precision, accuracy, and linearity. In presence of most common interferents, both test systems could lead to accurate creatinine results except for the existence of specified drugs. For dobutamine, the open test system showed better anti-interference performance than the matched system. CONCLUSION: This study provides referable opinions for clinical laboratory selection on the test system and a framework for future analogous studies based on different test systems.

Blind Remote Sensing Image Deblurring Based on Overlapped Patches' Non-Linear Prior.

The remote sensing imaging environment is complex, in which many factors cause image blur. Thus, without prior knowledge, the restoration model established to obtain clear images can only rely on the observed blurry images. We still build the prior with extreme pixels but no longer traverse all pixels, such as the extreme channels. The features are extracted in units of patches, which are segmented from an image and partially overlap with each other. In this paper, we design a new prior, i.e., overlapped patches' non-linear (OPNL) prior, derived from the ratio of extreme pixels affected by blurring in patches. The analysis of more than 5000 remote sensing images confirms that OPNL prior prefers clear images rather than blurry images in the restoration process. The complexity of the optimization problem is increased due to the introduction of OPNL prior, which makes it impossible to solve it directly. A related solving algorithm is established based on the projected alternating minimization (PAM) algorithm combined with the half-quadratic splitting method, the fast iterative shrinkage-thresholding algorithm (FISTA), fast Fourier transform (FFT), etc. Numerous experiments prove that this algorithm has excellent stability and effectiveness and has obtained competitive processing results in restoring remote sensing images.

The Growth, Lipid Accumulation and Fatty Acid Profile Analysis by Abscisic Acid and Indol-3-Acetic Acid Induced in Chlorella sp. FACHB-8.

Microalgae are considered a promising source for biodiesel. The addition of plant hormone can exert a significant impact on the production of microalgae biomass and lipid accumulation. Nevertheless, the response of microalgae cells to hormones is species- or strain-dependent. It remains controversial which genes involved in strong increase of fatty acids production in response to abscisic acid (ABA) in Chlorella sp. FACHB-8 strain. We investigated cell growth, lipid accumulation, and fatty acid composition when ABA and indol-3-acetic acid (IAA) were used in the growth medium of Chlorella sp. FACHB-8. The four treatments, including 5 mg/L IAA (E1), 10 mg/L IAA (E2), 10 mg/L ABA (E3), the combination of 5 mg/L IAA and 5 mg/L ABA (E4), were found to increase cell growth, but only 10 mg/L ABA treatment could enhance the lipid accumulation. The fatty acid profile was changed by the addition of ABA, making fatty acids afflux from polyunsaturated fatty acids to monounsaturated and saturated fatty acids, which were suitable for diesel application. Furthermore, a transcriptome analysis was conducted, unraveling the differentially expressed genes enriched in fatty acid biosynthesis, fatty acid metabolism, and biosynthesis of the unsaturated fatty acid pathway in response to ABA. Our results clarified the correlation of fatty acid synthesis-related genes and fatty acid profiles, helping understand the potential response mechanism of Chlorella sp. FACHB-8 strain respond to ABA treatment.

Identification of the potential novel biomarkers as susceptibility gene for Wilms tumor.

BACKGROUND: Wilms tumor (WT) is the most common malignant renal tumor in children. The aim of this study was to identify potential susceptibility gene of WT for better prognosis. METHODS: Weighted gene coexpression network analysis is used for the detection of clinically important biomarkers associated with WT. RESULTS: In the study, 59 tissue samples from National Cancer Institute were pretreated for constructing gene co-expression network, while 224 samples also downloaded from National Cancer Institute were used for hub gene validation and module preservation analysis. Three modules were found to be highly correlated with WT, and 44 top hub genes were identified in these key modules eventually. In addition, both the module preservation analysis and gene validation showed ideal results based on other dataset with 224 samples. Meanwhile, Functional enrichment analysis showed that genes in module were enriched to sister chromatid cohesion, cell cycle, oocyte meiosis. CONCLUSION: In summary, we established a gene co-expression network to identify 44 hub genes are closely to recurrence and staging of WT, and 6 of these hub genes was closely related to the poor prognosis of patients. Our findings revealed that those hub genes may be used as potential susceptibility gene for clinical diagnosis and prognosis of this tumor.

GlyinsRNA: a webserver for predicting glycosylation sites on small RNAs.

Versatile RNA modifications play important roles in post-transcriptional regulations of gene expression, among which glycosylation modifications on small RNAs emerge as a novel clade whose characteristics need further interrogations. Here, we demonstrated that the sequence pattern around RNA glycosylation sites was not random and could be exploited for glycosylation site prediction. A machine learning predictor, GlyinsRNA, which integrated multiple RNA sequence representation encodings, was established. GlyinsRNA achieved AUROC (area under the receiver operating characteristic curve) of 0.7933 and 0.7979 in five-fold cross-validation and independent tests, respectively. GlyinsRNA was implemented as an online webserver, where both the predicted glycosylation sites and the overrepresented RNA-binding protein (RBP)-related motifs were annotated to facilitate the users. GlyinsRNA webserver is freely available at http://www.rnanut.net/glyinsrna.

An isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for aldosterone measurement in human plasma.

Accurate quantitation of aldosterone is clinically important in standardized testing for primary aldosteronism. The results are often variable when performed by clinical immunoassays. To standardize and ensure the accuracy of clinical systems, reference measurement procedures (RMPs) with higher metrological order are required. A simple and reliable isotope dilution LC-IDMS/MS-based measurement procedure for human plasma aldosterone has been developed. This method involved plasma spiked with a deuterium-labelled internal standard, equilibrated for 0.5 h, and extracted by liquid-liquid extraction (LLE) without derivatization. Aldosterone and its structural analogues were baseline separated with a C18-packed UHPLC column with gradient elution within 7 min. The signal intensity variability and measurement imprecision were reduced by bracketing calibration during plasma aldosterone value assignment. The limit of detection (LoD) was 19.4 pmol/L with a signal-to-noise ratio (S/N) > 3. The lowest limit of quantification (LLoQ) was 27.7 pmol/L (S/N > 10 and CV < 10.0%). LLE was performed with 1 mL of n-hexane/ethyl acetate (3:2, v/v), and the extraction recovery was determined to be 92.15 ± 3.54%. The imprecisions were ≤ 3.18% for samples at 124.8, 867.0, and 2628.5 pmol/L. The recoveries were 98.11-101.61%. The relative bias between this candidate RMP and the established RMP was 2.76-1.89%. The linearity response ranged from 27.7 to 2774.4 pmol/L with R2 = 0.999. The method performance met the requirements of RMPs (≤ 5% total CV and ≤ 3% bias). Furthermore, the developed method was applied to evaluate immunoassays through 41 patient sample comparisons. The calibration and measurement capability (CMC) of this method were also evaluated by measuring these samples. The candidate RMP can serve as an accurate reference baseline for routine methods and can be used for value assignment for reference materials. Selected ion chromatograms by LC-MS/MS using a C18 column for aldosterone and its structural analogues.

Retracted: MicroRNA-495 Inhibits Gastric Cancer Cell Migration and Invasion Possibly via Targeting High Mobility Group AT-Hook 2 (HMGA2).

This publication has been retracted by the Editor due to the identification of non-original figure images and manuscript content that raise concerns regarding the credibility and originality of the study and the manuscript. Reference: Huashe Wang, Zhipeng Jiang, Honglei Chen, Xiaobin Wu, Jun Xiang, Junsheng Peng. MicroRNA-495 Inhibits Gastric Cancer Cell Migration and Invasion Possibly via Targeting High Mobility Group AT-Hook 2 (HMGA2). Med Sci Monit, 2017; 23: 640-648. DOI: 10.12659/MSM.898740.

Psychological characteristics of parents in a Paediatric Outpatient during the SARS-CoV-2 Epidemic.

With the outbreak and spread of the novel coronavirus (SARS-CoV-2), there has been a significant increase in the number of children infected, and some severe infection cases and neonatal cases have been reported. The parents or other family members who come to our paediatric clinic inevitably experience panic, tension and anxiety. The generation of these emotions has seriously affected the normal order of outpatient treatment and has led to many children not receiving an accurate diagnosis or proper treatment. This situation is not conducive to the control of the epidemic or the children's physical and mental health. Through summarizing parents' behaviours and emotional characteristics during the epidemic period, we hope to develop relevant coping and nursing strategies to ensure better control of the epidemic and to protect the physical and mental health of children.

Computational screening of potential regulators for mRNA-protein expression level discrepancy.

Complicated post-transcriptional and translational regulation processes contribute to the expression discrepancy between mRNA and protein in many tissues, but the underlying mechanisms have not been fully understood. In this study, we assessed to what extent and which RNA binding proteins (RBPs) contribute to mRNA-protein expression discrepancy. To this end, we exploited the RNA-seq transcriptome data and corresponding quantitative proteome data from the same set of human healthy tissues to estimate the mRNA-protein expression discrepancy, and observed that a considerable fraction of genes show obvious difference in expression rankings between transcriptome and proteome. We further assembled the latest CLIP-seq datasets from POSTAR2, ENCODE and GEO to map the binding profiles of known RBPs. A logistic regression model based on the RBP-binding features was established, which could predict the mRNA-protein expression discrepancy with acceptable performance. Finally, by applying two different feature selection methods on this logistic regression model, we identified a consensus set of known and putative translation regulators which may account for the expression level discrepancy, such as G3BP1, DGCR8, LARP4B, EIF4A3 and FXR2.

Genome-wide systematic characterization of the HAK/KUP/KT gene family and its expression profile during plant growth and in response to low-K+ stress in Saccharum.

BACKGROUND: Plant genomes contain a large number of HAK/KUP/KT transporters, which play important roles in potassium uptake and translocation, osmotic potential regulation, salt tolerance, root morphogenesis and plant development. Potassium deficiency in the soil of a sugarcane planting area is serious. However, the HAK/KUP/KT gene family remains to be characterized in sugarcane (Saccharum). RESULTS: In this study, 30 HAK/KUP/KT genes were identified in Saccharum spontaneum. Phylogenetics, duplication events, gene structures and expression patterns were analyzed. Phylogenetic analysis of the HAK/KUP/KT genes from 15 representative plants showed that this gene family is divided into four groups (clades I-IV). Both ancient whole-genome duplication (WGD) and recent gene duplication contributed to the expansion of the HAK/KUP/KT gene family. Nonsynonymous to synonymous substitution ratio (Ka/Ks) analysis showed that purifying selection was the main force driving the evolution of HAK/KUP/KT genes. The divergence time of the HAK/KUP/KT gene family was estimated to range from 134.8 to 233.7 Mya based on Ks analysis, suggesting that it is an ancient gene family in plants. Gene structure analysis showed that the HAK/KUP/KT genes were accompanied by intron gain/loss in the process of evolution. RNA-seq data analysis demonstrated that the HAK/KUP/KT genes from clades II and III were mainly constitutively expressed in various tissues, while most genes from clades I and IV had no or very low expression in the tested tissues at different developmental stages. The expression of SsHAK1 and SsHAK21 was upregulated in response to low-K+ stress. Yeast functional complementation analysis revealed that SsHAK1 and SsHAK21 could rescue K+ uptake in a yeast mutant. CONCLUSIONS: This study provided insights into the evolutionary history of HAK/KUP/KT genes. HAK7/9/18 were mainly expressed in the upper photosynthetic zone and mature zone of the stem. HAK7/9/18/25 were regulated by sunlight. SsHAK1 and SsHAK21 played important roles in mediating potassium acquisition under limited K+ supply. Our results provide valuable information and key candidate genes for further studies on the function of HAK/KUP/KT genes in Saccharum.

RNA N6-methyladenosine demethylase FTO promotes breast tumor progression through inhibiting BNIP3.

BACKGROUND: N6-methyladenosine (m6A) modification is the most pervasive modification in mRNA, and has been considered as a new layer of epigenetic regulation on mRNA processing, stability and translation. Despite its functional significance in various physiological processes, the role of the m6A modification involved in breast cancer is yet fully understood. METHODS: We used the m6A-RNA immunoprecipitation sequencing to identify the potential targets in breast cancer. To determine the underlying mechanism for the axis of FTO-BNIP3, we performed a series of in vitro and in vivo assays in 3 breast cancer cell lines and 36 primary breast tumor tissues and 12 adjunct tissues. RESULTS: We showed that FTO, a key m6A demethylase, was up-regulated in human breast cancer. High level of FTO was significantly associated with lower survival rates in patients with breast cancer. FTO promoted breast cancer cell proliferation, colony formation and metastasis in vitro and in vivo. We identified BNIP3, a pro-apoptosis gene, as a downstream target of FTO-mediated m6A modification. Epigenetically, FTO mediated m6A demethylation in the 3'UTR of BNIP3 mRNA and induced its degradation via an YTHDF2 independent mechanism. BNIP3 acts as a tumor suppressor and is negatively correlated with FTO expression in clinical breast cancer patients. BNIP3 dramatically alleviated FTO-dependent tumor growth retardation and metastasis. CONCLUSIONS: Our findings demonstrate the functional significance of the m6A modification in breast cancer, and suggest that FTO may serve as a novel potential therapeutic target for breast cancer.

Blue laser imaging with acetic acid enhancement improved the detection rate of gastric intestinal metaplasia.

Our aim was to evaluate the ability of blue laser imaging (BLI) combined with acetic acid (BLI-AA) to detect gastric intestinal metaplasia (GIM). Participants undergoing gastroscopy from July 2017 to February 2018 in our hospital were enrolled prospectively. The abilities of white light imaging endoscopy, BLI endoscopy, and BLI-AA to detect GIM were compared. One hundred six patients undergoing gastroscopy met the inclusion criteria. GIM was diagnosed in 41 patients. For BLI-AA, the sensitivity, specificity, positive predictive, and negative predictive values were 85.4%, 84.6%, 77.8%, and 90.2% respectively. The diagnostic accuracy rate for BLI-AA was 84.9%, which was higher than that of white light imaging endoscopy and BLI endoscopy. For target biopsy, the GIM detection rate for the BLI-AA mode was significantly higher (77.8%, 105/135) than that for the BLI mode (58.3%, 84/144) or the white light endoscopy mode (40.4%, 57/141) (p < 0.05). BLI-AA is an efficient and simple method for the detection of GIM.

Predictability of gastric intestinal metaplasia by patchy lavender color seen on linked color imaging endoscopy.

We aimed to investigate the ability of linked color imaging (LCI) versus white light endoscopy (WLE) to detect gastric intestinal metaplasia (GIM). One hundred and seven participants who underwent upper gastrointestinal endoscopy were included. Under WLE endoscopy, biopsies were performed on any suspected abnormal mucosal changes. Under LCI endoscopy, we tested whether the specific color feature of patchy lavender color (PLC) pathologically indicated GIM. Biopsies were randomly performed in participants who had neither PLC nor suspected lesions. The detection abilities of LCI and WLE were assessed by comparison of histological and endoscopic findings. A total of 41 participants had histological GIM. The total diagnostic accuracy rate for GIM by LCI was 79.44%, higher than that of WLE (40.19%) (P < 0.001). Moreover, LCI with targeted biopsies showed a significantly increased ability to detect GIM (P < 0.001). PLC observed in the gastric mucosa on LCI can guide endoscopic biopsies and increase the detection rate of GIM. Thus, LCI could be a good tool for detecting GIM. ClinicalTrials.gov Identifier: ChiCTR-DDD-17011326).

Toosendanin induces caspase-dependent apoptosis through the p38 MAPK pathway in human gastric cancer cells.

Although many advances have been made in the treatment of gastric cancer (GC), numerous difficulties, such as the emergence of chemo-drug resistance, continue to lead to disappointing GC prognoses. Thus, novel alternative strategies are urgently needed. The use of natural products could be a viable option to treat GC. Toosendanin (TSN) is a triterpenoid derived from the bark of Melia toosendanin Sieb. et Zucc that has been shown to be highly cytotoxic to multiple cancer cells. As the underlying impact of TSN on GC and its molecular mechanism remain poorly understood, in this study, we performed a series of experiments involving the use of TSN to treat GC cells. In the present study, we showed that TSN suppressed cell viability, inhibited cell proliferation by causing G1/S arrest and induced caspase-dependent apoptosis in AGS and HGC-27 cells. The possible mechanism of TSN-induced apoptosis may be associated with the activation of the p38 MAPK pathway. These results demonstrated the potential of TSN as a promising therapeutic compound to treat gastric cancer.

Interference of Thalassemias on Hemoglobin A1c Measurement by Ion-Exchange High-Performance Liquid Chromatography Method Tosoh HLC-723 G8.

BACKGROUND: The presence of hemoglobinopathies could interfere with some assays for Hemoglobin A1c (HbA1c) measurement; therefore, the effect of thalassemia on ion-exchange high-performance liquid chromatography (IEHPLC) method Tosoh HLC-723 G8 (Tosoh G8) was evaluated. METHODS: A total of 43 normal controls and 101 thalassemia patients were quantified by Premier Hb9210 and Tosoh G8 (variant-mode) systems. At the same time, 7 normal controls and 8 thalassemia patients were confirmed by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) reference method for verification. RESULTS: For normal controls, the HbA1c values of Tosoh G8 system (y) showed great correlation and agreement with those from Premier Hb9210 (x) (y = 0.9688x + 0.2151, r = 0.9951; mean difference 0.02 ± 0.30%), and no significant relative bias above 7% was observed; the HbA1c values obtained by Tosoh G8 were consistent with the IFCC targets (relative bias < ± 6%) in all of the samples. However, for thalassemia, the correlation between Tosoh G8 (y) and Premier Hb9210 (x) became relatively low (y = 0.8079x + 1.2897, r = 0.7780); the HbA1c values of 91.1% of the samples (92/101) obtained by Tosoh G8 were higher than those by Premier Hb9210 (mean difference 0.33 ± 0.48%) and a significant positive bias above 7% was noticed in 43.3% (45/101) thalassemia patients; when compared with the IFCC targets, the 87.5% (7/8) relative bias was > ± 6%. CONCLUSIONS: Thalassemia could directly affect the measurement of HbA1c using the IE-HPLC method Tosoh G8 and the clinical laboratorial staff should pay close attention.