Improved Dye Survival in Expansion Microscopy through Stabilizer-Conjugated Linkers.

Expansion microscopy (ExM) has been widely used to detect biomolecules in cultured cells and tissue samples due to its enablement of super resolution imaging with conventional microscopes, via physical expansion of samples. However, reaction conditions inherent to the process bring about strong fluorescent signal loss during polymerization and digestion and thus limit the brightness of the signal obtained post expansion. Here, we explore the impact of stabilizer-containing organic fluorophores in ExM, as a mitigation strategy for this radical-induced dye degradation. Through direct conjugation of 4-nitrophenylalanine (NPA) to our previously developed trifunctional reagents, we validate and demonstrate that these multifunctional linkers enable visualization of different organelles with improved fluorescent intensity, owning to protection of the dyes to radical induced degradation as well as to photoprotection upon imaging. At this point, we cannot disentangle the relative contribution of both mechanisms. Furthermore, we report anchoring linkers that allow straightforward application of NPA or Trolox to commercially available fluorophore-conjugated antibodies. We show that these anchoring linkers enable complete retention of biological targets while increasing fluorophore photostability. Our results provide guidance in exploring these stabilizer-modified agents in ExM and methods for increased signal survival through the polymerization steps of the ExM protocols.

Bionic Covert Underwater Acoustic Communication Based on Time-Frequency Contour of Bottlenose Dolphin Whistle.

In order to meet the requirements of communication security and concealment, as well as to protect marine life, bionic covert communication has become a hot research topic for underwater acoustic communication (UAC). In this paper, we propose a bionic covert UAC (BC-UAC) method based on the time-frequency contour (TFC) of the bottlenose dolphin whistle, which can overcome the safety problem of traditional low signal-noise ratio (SNR) covert communication and make the detected communication signal be excluded as marine biological noise. In the proposed BC-UAC method, the TFC of the bottlenose dolphin whistle is segmented to improve the transmission rate. Two BC-UAC schemes based on the segmented TFC of the whistle, the BC-UAC scheme using the whistle signal with time-delay (BC-UAC-TD) and the BC-UAC scheme using the whistle signal with frequency-shift (BC-UAC-FS), are addressed. The original whistle signal is used as a synchronization signal. Moreover, the virtual time reversal mirror (VTRM) technique is adopted to equalize the channel for mitigating the multipath effect. The performance of the proposed BC-UAC method, in terms of the Pearson correlation coefficient (PCC) and bit error rate (BER), is evaluated under simulated and measured underwater channels. Numerical results show that the proposed BC-UAC method performs well on covertness and reliability. Furthermore, the covertness of the bionic modulated signal in BC-UAC-TD is better than that of BC-UAC-FS, although the reliability of BC-UAC-FS is better than that of BC-UAC-TD.

Effects of Postmortem Hemolysis and Ultrafiltration on Creatinine Detection Results.

OBJECTIVES: To investigate the interference of postmortem hemolysis on the detection of creatinine and whether ultrafiltration can reduce the interference. METHODS: A total of 33 non-hemolyzed whole blood samples from the left heart were collected. Hemolyzed samples with 4 hemoglobin mass concentration gradients H1-H4 were artificially prepared. Ultrafiltration was performed on each hemolyzed sample. Creatinine concentrations in non-hemolyzed serum (baseline serum), hemolyzed samples and ultrafiltrate were detected. Bias (B), Pearson correlation and receiver operator characteristic (ROC) of baseline creatinine concentration between before and after ultrafiltration were analyzed. RESULTS: As the hemoglobin mass concentration increased, B of the hemolyzed samples in the H1-H4 groups gradually increased, the |B| was 2.41(0.82, 8.25)-51.31(41.79, 188.25), reaching a maximum of 589.06%, and there was no statistically significant between the creatinine concentration and the baseline creatinine concentration (P=0.472 7, r=0.129 5). After ultrafiltration of hemolyzed samples, the interference of creatinine concentration in ultrafiltrate was significantly reduced, the |B| was 5.32(2.26, 9.22)-21.74(20.06, 25.58), reaching a maximum of 32.14%, and there was a positive correlation with baseline creatinine concentration (P<0.05, r=0.918 2). In the hemolyzed samples of H3 and H4 groups, there were 7 false-positive samples and 1 false-negative sample; in the ultrafiltrate samples, there were no false-positive sample and 1 false-negative sample. ROC analysis results showed the hemolyzed samples were lack of diagnostic value (P=0.117 5). CONCLUSIONS: The postmortem hemolysis significantly interferes creatinine detection results of blood samples, ultrafiltration can reduce hemolysis-induced interference in postmortem creatinine detection.

Interference of hemolysis on the postmortem biochemical analysis of IgE by ECLIA.

Forensic diagnosis of anaphylactic shock is a challenging task in forensic practice due to the lack of characteristic morphological changes. Postmortem analysis of serum IgE can provide helpful information for determining anaphylaxis. However, postmortem serum always suffers from hemolysis. To investigate the interference of hemolysis on postmortem analysis of total IgE by electrochemiluminescent immunoassay (ECLIA) and verify the suitability of the commercially available ECLIA kit for postmortem hemolyzed blood with the dilution-correction method, different levels of hemolyzed serum were prepared to evaluate the interference of hemolysis. A linear regression analysis was then performed on the concentration of total IgE in the completely hemolyzed blood and the corresponding serum. Our results indicated that hemolysis negatively interfered with the total IgE analysis by ECLIA and the interference (|Bias%|) increased with increasing levels of hemolysis. After controlling for |Bias%| by dilution, the test concentration of total IgE in the completely hemolyzed blood was still significantly lower than that in the serum (P < 0.05) and resulted in eight false-negative cases. A strong correlation was observed between the test concentration of total IgE in the completely hemolyzed blood and that in the serum (r = 0.983). After correction by the regression formula, the corrected concentration revealed no significant differences and exhibited the same diagnostic ability, compared with the serum total IgE concentration. These results indicate that the completely hemolyzed blood is not recommended for postmortem analysis of total IgE directly. The dilution-correction method might have potential utility in forensic practice for evaluating serum total IgE concentrations.

AIN-93 Diet as an Alternative Model to Lieber-DeCarli Diet for Alcoholic Cardiomyopathy.

BACKGROUND: The Lieber-DeCarli alcoholic liquid diet is a classical method for establishing animal models of alcoholic cardiomyopathy (ACM). No study has reported whether the AIN-93 diet, which is widely used as a standard diet for both long-term and short-term studies with laboratory animals, could be used to construct the ACM animal model. The present study intended to investigate whether the AIN-93 diet could be used to establish a mouse ACM model. METHODS: Twenty-four C57BL/6 male mice were randomly divided into 4 equally sized groups. In ethanol (EtOH)-fed groups, mice were fed a 4%-EtOH (w/v, 28% of total calories) alcoholic liquid diet of Lieber-DeCarli or the AIN-93 diet for chronic alcohol exposure for 180 days. In control-fed groups, mice were fed with non-EtOH liquid diets with the same calories as EtOH-fed groups. Morphological observations of the hearts and molecular investigation of the brain natriuretic peptide (BNP) were carried out by echocardiography, hematoxylin and eosin (H&E) and immunohistochemistry (IHC) staining, real-time quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay. RESULTS: Echocardiography showed that mice fed with either the 4%-EtOH Lieber-DeCarli diet or the 4%-EtOH AIN-93 diet had dilated ventricles and poor cardiac function. IHC staining of BNP, qPCR of BNP mRNA, and plasma concentration of BNP showed an up-regulated expression in mice fed with both the 4%-EtOH Lieber-DeCarli and 4%-EtOH AIN-93 diets. Less fatty liver was also observed in mice fed the AIN-93 alcoholic diet than those fed the Lieber-DeCarli alcoholic diet. CONCLUSIONS: The AIN-93 alcoholic liquid diet can be used to establish ACM animal models, as with the conventional Lieber-DeCarli alcoholic liquid diet.

BNP and NT-proBNP as Diagnostic Biomarkers for Cardiac Dysfunction in Both Clinical and Forensic Medicine.

Currently, brain natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) are widely used as diagnostic biomarkers for heart failure (HF) and cardiac dysfunction in clinical medicine. They are also used as postmortem biomarkers reflecting cardiac function of the deceased before death in forensic medicine. Several previous studies have reviewed BNP and NT-proBNP in clinical medicine, however, few articles have reviewed their application in forensic medicine. The present article reviews the biological features, the research and application status, and the future research prospects of BNP and NT-proBNP in both clinical medicine and forensic medicine, thereby providing valuable assistance for clinicians and forensic pathologists.

Diagnostic Roles of Postmortem cTn I and cTn T in Cardiac Death with Special Regard to Myocardial Infarction: A Systematic Literature Review and Meta-Analysis.

BACKGROUND: Cardiac troponin I (cTn I) and cardiac troponin T (cTn T) are currently widely used as diagnostic biomarkers for myocardial injury caused by ischemic heart diseases in clinical and forensic medicine. However, no previous meta-analysis has summarized the diagnostic roles of postmortem cTn I and cTn T. The aim of the present study was to meta-analyze the diagnostic roles of postmortem cTn I and cTn T for cardiac death in forensic medicine, present a systematic review of the previous literature, and determine the postmortem cut-off values of cTn I and cTn T. METHODS: We searched multiple databases for the related literature, performed a meta-analysis to investigate the diagnostic roles of postmortem cardiac troponins, and analyzed the receiver operating characteristic (ROC) curve to determine their postmortem cut-off values. RESULTS AND CONCLUSIONS: The present meta-analysis demonstrated that postmortem cTn I and cTn T levels were increased in pericardial fluid and serum in cardiac death, especially in patients with acute myocardial infarction (AMI). We determined the postmortem cut-off value of cTn I in the pericardial fluid at 86.2 ng/mL, cTn I in serum at 9.5 ng/mL, and cTn T in serum at 8.025 ng/mL.

The Expression of BNP, ET-1, and TGF-ß1 in Myocardium of Rats with Ventricular Arrhythmias.

Ventricular arrhythmia (VA) is a major component of sudden cardiac death (SCD). To investigate the expression of brain natriuretic peptide (BNP), endothelin-1 (ET-1), and transforming growth factor-beta 1 (TGF-ß1) during VA, we established a rat model of VA induced by BaCl2 solution through a microinjector pump. PD142893 (ET-1 receptor blocker) and SB431542 (TGF-ß1 receptor type I blocker) were used to explore the effect of ET-1 and TGF-ß1 on BNP expression in the myocardium after VA. BNP, ET-1, and TGF-ß1 in rat myocardium were assayed by western blot and immunohistochemical staining for proteins, and real-time quantitative polymerase chain reaction for mRNAs. We found increased expression of BNP and ET-1 in rat myocardium that was associated with the duration of VA. However, TGF-ß1 protein expression remained unchanged. Such early increases in BNP and ET-1 may be attributed to fatal arrhythmias associated with SCD, suggesting these may be novel biomarkers of this disease. After intraperitoneal injection of PD142893 and SB431542, respectively, BNP was downregulated in the myocardium of the left ventricle; however, this was abrogated by co-application of the two inhibitors. These results suggested that both ET-1 and TGF-ß1, by specifically binding to their receptors, might be involved in the myocardial synthesis of BNP during VA in vivo.

Clinical and historical infection of Tacheng tick virus 2: A retrospective investigation.

BACKGROUND: Tacheng tick virus 2 (TcTV-2) is an emerging tick-borne virus belonging to the genus Uukuvirus in the family Phenuiviridae. Initially isolated in 2019 from a patient in Xinjiang Uygur Autonomous Region (XUAR), northwestern China, who developed fever and headache after a tick bite, TcTV-2 was concurrently molecularly detected in hard ticks across various countries, including China, Kazakhstan, Romania, and Turkey. This study conducted a retrospective epidemiological investigation of TcTV-2 infection. METHODOLOGY: In this retrospective cohort study, we collected samples from 47 tick-bitten patients, 984 herdsmen, 7 Asian badgers, 13 red foxes, and 168 Hyalomma asiaticum tick egg batches. Patients' samples were primarily analyzed by using high-throughput sequencing, targeting the V3-V4 region of the bacterial 16S rRNA gene and viral cDNA libraries. Typical tick-borne pathogens were further confirmed using RT-PCR and detected in Asian badgers, red foxes and Hy. asiaticum tick egg batches. We also conducted enzyme-linked immunosorbent assay (ELISA) to detected specific IgM and IgG antibodies against TcTV-2 in herdsmen. Phylogenetic analysis was performed to genetically characterize TcTV-2 detected in this study. PRINCIPAL FINDINGS: TcTV-2 was detected in various samples, including blood, urine, and throat swabs from 12.77% (6/47) tick-bitten patients. It was found in blood samples of 14.29% (1/7) of wild badgers, 7.69% (1/13) of red foxes, and 13.69% (23/168) of Hy. asiaticum egg batches. Furthermore, ELISA results revealed that 9.55% (94/984) of the serum samples (34 from males and 60 from females) were tested positive for TcTV-2-specific IgG, while 2.95% (29/984, 7 males and 22 females) showed positivity for TcTV-2-specific IgM. Additionally, 1.02% (10/984, 4 males and 6 females) of the sera tested positive for both TcTV-2-specific IgM and IgG. Phylogenetic analysis indicated that the TcTV-2 strains detected in this study were genetically similar, regardless of their origin and host species. CONCLUSIONS: Clinical symptoms of TcTV-2 infection in patients are nonspecific, with common symptoms including headache, fever, asthenia, vomiting, myalgia, rash, and meningitis-like signs. TcTV-2 can be detected in blood, urine, and throat swab samples of infected patients. Among local herdsmen, 9.55% tested positive for TcTV-2-specific IgG and 2.95% for TcTV-2-specific IgM. Importantly, TcTV-2 can be transovarially transmitted in Hy. asiaticum ticks, and the Asian badgers and red foxes are potential reservoirs of TcTV-2.

Development of Inhibitors, Probes, and PROTAC Provides a Complete Toolbox to Study PARK7 in the Living Cell.

The integration of diverse chemical tools like small-molecule inhibitors, activity-based probes (ABPs), and proteolysis targeting chimeras (PROTACs) advances clinical drug discovery and facilitates the exploration of various biological facets of targeted proteins. Here, we report the development of such a chemical toolbox for the human Parkinson disease protein 7 (PARK7/DJ-1) implicated in Parkinson's disease and cancers. By combining structure-guided design, miniaturized library synthesis, and high-throughput screening, we identified two potent compounds, JYQ-164 and JYQ-173, inhibiting PARK7 in vitro and in cells by covalently and selectively targeting its critical residue, Cys106. Leveraging JYQ-173, we further developed a cell-permeable Bodipy probe, JYQ-196, for covalent labeling of PARK7 in living cells and a first-in-class PARK7 degrader JYQ-194 that selectively induces its proteasomal degradation in human cells. Our study provides a valuable toolbox to enhance the understanding of PARK7 biology in cellular contexts and opens new opportunities for therapeutic interventions.

Next-Generation Sequencing of the Complete Huaibei Grey Donkey Mitogenome and Mitogenomic Phylogeny of the Equidae Family.

The Huaibei grey donkey (HGD) is an endangered species and a vital native breed in Anhui Province, China. However, its complete mitogenome, phylogeny, and maternal origin remain unclear. The objectives of this study were to detect the genetic diversity of the HGD and investigate its phylogenetic relationship with other breeds to inform conservation management. The complete mitogenome of the HGD was sequenced through next-generation sequencing, and the most variable region in the mitochondrial DNA displacement-loop (D-loop) was amplified via a polymerase chain reaction (PCR). Next, we used the median-joining network (MJN) to calculate the genetic relationships among populations and the neighbor-jointing method to build a phylogenetic tree and speculate as to its origin. The results showed that the mitogenome contains 22 tRNAs, 2 rRNAs, 13 PCGs, and 1 D-loop region. Analyzing the D-loop region of the HGDs, we identified 23 polymorphic sites and 11 haplotypes. The haplotype and nucleotide diversity were 0.87000 (Hd) and 0.02115 (Pi), respectively. The MJN analysis indicated that the HGD potentially has two maternal lineages, and phylogenetic analysis indicated that the Somali lineage could be the most probable domestication center for this breed. Therefore, our mitogenome analysis highlights the high genetic diversity of the HGD, which may have originated from the Somali wild ass, as opposed to the Asian wild ass. This study will provide a useful resource for HGD conservation and breeding.

Mitochondrial genome and phylogenetic analysis of Chaohu duck.

A complete mitochondrial genome sequence is important for the accurate determination of phylogenetic relationships. Chaohu duck is a dominant native breed in Anhui Province, China. We aimed to ascertain the complete mitochondrial genome sequence of Chaohu duck via high-throughput sequencing and primer walking. Phylogenetic analysis of Chaohu duck was performed following Kimura 2-parameter model. The total length of the mitogenome was 16,597 bp, and comprised 29.2 %A, 22.2 % T, 32.8 % C, and 15.8 % G. It included 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes and a control region (D-loop). Furthermore, the haplotype diversity and nucleotide diversity values were 0.9028(Hd) and 0.01162(Pi) respectively. This indicates that Chaohu duck has high population diversity. Twenty-two haplotypes were identified in sixty Chaohu ducks which were divided into two haplogroups. Therefore, we inferred that Chaohu duck may originate from Anas platyrhynchos, and was influenced by Anas poecilorhyncha during evolution. Our results provide mitochondrial genome information for further studies on Chaohu ducks and lays a foundation for germplasm resources conservation.

Stepwise formation of a chemodynamic therapy agent of {Cu8} macrocyclic complex recognized by iodide ions.

An atomically precise Cu(I) macrocyclic complex Cu8I was developed for chemodynamic therapy (CDT) research. The {Cu8} macrocyclic skeleton gradually forms with the selective recognition of iodide ions, and the monitoring of intermediate fragments during Cu8I formation using time-dependent electrospray ionization mass spectrometry indicates the following possible formation process: [Cu1] → [Cu2] → [Cu3] → [Cu4] → [Cu5I] → [Cu6I] → [Cu7I] → [Cu8I] when recognized by iodide ions. Furthermore, the Cu(I)-mediated Fenton-like reaction in Cu8I catalyzes the production of toxic ˙OH from H2O2, which results in efficient tumor suppression.

Trifunctional Linkers Enable Improved Visualization of Actin by Expansion Microscopy.

Expansion microscopy (ExM) revolutionized the field of super-resolution microscopy by allowing for subdiffraction resolution fluorescence imaging on standard fluorescence microscopes. However, it has been found that it is hard to visualize actin filaments efficiently using ExM. To improve actin imaging, multifunctional molecules have been designed with moderate success. Here, we present optimized methods for phalloidin conjugate grafting that have a high efficiency for both cellular and tissue samples. Our optimized strategy improves anchoring and signal retention by ∼10 times. We demonstrate the potential of optimized trifunctional linkers (TRITON) for actin imaging in combination with immunolabeling using different ExM protocols. 10X ExM of actin labeled with optimized TRITON enabled us to visualize the periodicity of actin rings in cultured hippocampal neurons and brain slices by Airyscan confocal microscopy. Thus, TRITON linkers provide an efficient grafting method, especially in cases in which the concentration of target-bound monomers is insufficient for high-quality ExM.

Characteristics of the complete mitochondrial genome of Douhua chicken (Gallus gallus) and phylogenetic considerations.

Douhua chicken is a unique local breed from Anhui Province, China. This study aimed to illustrate the Douhua chicken mitogenome and clarify its phylogenetic status by sequencing and annotating the complete mitochondrial genome using high-throughput sequencing and primer walking. Phylogenetic analysis through the Kimura 2-parameter model indicated the maternal origin of Douhua chicken. The results revealed that the mitochondrial genome is a closed circular molecule (16,785 bp) that consists of 13 protein-coding genes, 22 transfer RNA (tRNA) coding genes, two ribosomal RNA (rRNA) coding genes, and a control region. The base composition of the Douhua chicken mitogenome contains 30.3% A, 23.7% T, 32.5% C, and 13.5% G, and the haplotype and nucleotide diversity values are 0.829 (Hd) and 0.00441 (Pi), respectively. Furthermore, 10 haplotypes of D-loop sequences among 60 Douhua chickens were identified and distributed into four haplogroups (A, C, D, and E). Overall, the result of the present study indicates that Douhua chicken may have originated from Gallus gallus, and this process was influenced by Gallus gallus spadiceus, Gallus gallus murghi, and Gallus gallus bankiva. This study provides novel mitogenome data to support further phylogenetic and taxonomic studies on Douhua chicken. Additionally, the findings of this study will provide deeper insights for identifying the genetic relationships among populations and tracing maternal origins based on phylogenetic considerations for use in studies on the geographic conservation, utilization, and molecular genetics of poultry species.

In the modern poultry industry, resources of native varieties have become major aspects. Douhua chicken is a medium-sized, slow-growing, and white-feathered local breed that represents a popular local chicken breed in Anhui Province, China. This breed is adaptable and exhibits important production traits and a stable inheritable characteristics, such as delicious meat and stable egg-laying performance. The present study aimed to provide a better understanding of the germplasm characteristics and phylogenetic relationships of Douhua chicken by analyzing its complete mitochondrial genome sequence and a describing its genomic composition, nucleotide composition, and gene structure. The present study provides theoretical support for the protection, development, and utilization of Douhua chicken resources. Additionally, this study provides new mitochondrial genome data to support further phylogenetic and taxonomic studies conducted on Douhua chicken.

Spotted fever group rickettsiae in hard ticks in eastern and southern Kazakhstan.

Infections with spotted fever group rickettsiae represent a worldwide health problem, characterized by persistent high fever, headache, and rash in humans, domestic animals, and wildlife. To date, the occurrence of Rickettsia species in hard ticks has not been thoroughly studied, especially in eastern and southern Kazakhstan. A total of 1,245 adult ticks, comprising 734 Dermacentor marginatus, 219 Hyalomma scupense, 144 Hyalomma asiaticum, 84 Hyalomma marginatum, 48 Rhipicephalus turanicus, and 16 Haemaphysalis erinacei, collected from East Kazakhstan, Abay, Jetsu, Almaty, Jambyl, South Kazakhstan and Qyzylorda oblasts of Kazakhstan, were used to screen rickettsial agents using molecular methods. Rickettsia raoultii, Rickettsia slovaca, Rickettsia aeschlimannii and Rickettsia heilongjiangensis were identified using sequencing, and 31.5% (392/1245) of ticks carried rickettsial agents. The difference in the natural landscapes explains the variety of the collected ticks and expands our knowledge of Rickettsia species and their geographical distribution in Kazakhstan. To the best of our knowledge, this study reports the first finding of R. heilongjiangensis in Kazakhstan.

Rickettsia aeschlimannii Infection in a Woman from Xingjiang, Northwestern China.

Tick-borne Rickettsia aeschlimannii infection in humans has been described in several countries. This is the first report of R. aeschlimannii in a woman who reported being bitten by ticks in Xingjiang, northwestern China. R. aeschlimannii infection was confirmed by molecular detection in blood and urine of the patient, who presented clinical symptoms of severe edema, partial necrosis, and monocytosis. R. aeschlimannii was also detected in Hyalomma asiaticum ticks around the patient's residence. Infections of spotted fever group Rickettsia species should be included in the differential diagnosis from other tick-borne diseases.

Chemical Toolkit for PARK7: Potent, Selective, and High-Throughput.

The multifunctional human Parkinson's disease protein 7 (PARK7/DJ1) is an attractive therapeutic target due to its link with early-onset Parkinson's disease, upregulation in various cancers, and contribution to chemoresistance. However, only a few compounds have been identified to bind PARK7 due to the lack of a dedicated chemical toolbox. We report the creation of such a toolbox and showcase the application of each of its components. The selective PARK7 submicromolar inhibitor with a cyanimide reactive group covalently modifies the active site Cys106. Installment of different dyes onto the inhibitor delivered two PARK7 probes. The Rhodamine110 probe provides a high-throughput screening compatible FP assay, showcased by screening a compound library (8000 molecules). The SulfoCy5-equipped probe is a valuable tool to assess the effect of PARK7 inhibitors in a cell lysate. Our work creates new possibilities to explore PARK7 function in a physiologically relevant setting and develop new and improved PARK7 inhibitors.

Increased serum calcium levels are associated with carotid atherosclerotic plaque in normocalcaemic individuals with type 2 diabetes.

BACKGROUND: Patients with type 2 diabetes mellitus (T2DM) have an elevated risk of atherosclerotic cardiovascular disease. Although previous data have suggested that serum calcium levels could be involved in T2DM and cardiovascular disease, whether this applies in T2DM patients with atherosclerosis remains unclear. This study therefore aimed to investigate the relationship between serum calcium levels within the physiological ranges and carotid atherosclerotic plaque in T2DM patients. METHODS: A total of 594 normocalcaemic in-patients with T2DM were recruited, of whom 231 had carotid atherosclerotic plaque. Serum calcium levels were measured and carotid ultrasonography was performed. RESULTS: Patients with plaque had significantly higher serum albumin-corrected calcium than those without plaque [9.02 (8.78-9.34) mg/dL versus 8.86 (8.66-9.06) mg/dL, p < 0.001]. As serum albumin-corrected calcium levels increased across tertiles, the percentage of plaque increased (27.6%, 35.5%, and 55.7%; p < 0.001). Logistic regression showed that serum albumin-corrected calcium levels were independently and positively correlated with the presence of plaque, but not parathyroid hormone levels. Compared with patients in the lowest serum calcium tertiles, the odds ratio for plaque in patients in the upper quartile was 2.47 (95% confidence interval 1.51-4.03, p < 0.001) after adjustment for potential confounders. CONCLUSION: Serum albumin-corrected calcium levels are elevated in patients with T2DM and carotid atherosclerotic plaques.