Proteomic analysis of plasma proteins from patients with cardiac rupture after acute myocardial infarction using TMT-based quantitative proteomics approach.

BACKGROUND: Cardiac rupture (CR) is a rare but catastrophic mechanical complication of acute myocardial infarction (AMI) that seriously threatens human health. However, the reliable biomarkers for clinical diagnosis and the underlying signaling pathways insights of CR has yet to be elucidated. METHODS: In the present study, a quantitative approach with tandem mass tag (TMT) labeling and liquid chromatography-tandem mass spectrometry was used to characterize the differential protein expression profiles of patients with CR. Plasma samples were collected from patients with CR (n = 37), patients with AMI (n = 47), and healthy controls (n = 47). Candidate proteins were selected for validation by multiple reaction monitoring (MRM) and enzyme-linked immunosorbent assay (ELISA). RESULTS: In total, 1208 proteins were quantified and 958 differentially expressed proteins (DEPs) were identified. The difference in the expression levels of the DEPs was more noticeable between the CR and Con groups than between the AMI and Con groups. Bioinformatics analysis showed most of the DEPs to be involved in numerous crucial biological processes and signaling pathways, such as RNA transport, ribosome, proteasome, and protein processing in the endoplasmic reticulum, as well as necroptosis and leukocyte transendothelial migration, which might play essential roles in the complex pathological processes associated with CR. MRM analysis confirmed the accuracy of the proteomic analysis results. Four proteins i.e., C-reactive protein (CRP), heat shock protein beta-1 (HSPB1), vinculin (VINC) and growth/differentiation factor 15 (GDF15), were further validated via ELISA. By receiver operating characteristic (ROC) analysis, combinations of these four proteins distinguished CR patients from AMI patients with a high area under the curve (AUC) value (0.895, 95% CI, 0.802-0.988, p < 0.001). CONCLUSIONS: Our study highlights the value of comprehensive proteomic characterization for identifying plasma proteome changes in patients with CR. This pilot study could serve as a valid foundation and initiation point for elucidation of the mechanisms of CR, which might aid in identifying effective diagnostic biomarkers in the future.

A fluorescence-based immunochromatographic assay using quantum dot-encapsulated nanoparticles for the rapid and sensitive detection of fetuin-B.

Coronary artery disease (CAD) is the leading cause of death worldwide. Earlier detection of CAD improves treatment outcomes and secondary prevention. The circulating fetuin-B protein is considered to be a promising biomarker for the early detection of CAD. However, a facile and reliable clinical test for fetuin-B is still lacking. Herein, we describe a reliable fluorescent biosensor for detecting fetuin-B in plasma that combines quantum dots-doped polystyrene nanoparticles with an immunochromatographic assay strip (QNPs-ICAS). The QNPs served as detection signals in the QNPs-ICAS sensor system, which was based on a double-antibody sandwich structure. Under optimum experimental conditions, the biosensor exhibited a broad linear range of 1-200 ng mL-1 and a low detection limit of 0.299 ng mL-1. Furthermore, the proposed immunosensor demonstrated high sensitivity, satisfactory selectivity, good reproducibility, and excellent recovery. Finally, the performance and applicability of our QNPs-based ICAS system were validated in clinical samples using a commercial ELISA kit with excellent correlations (r = 0.98451, n = 116). To conclude, the proposed sensor served as a rapid, sensitive, and accurate method for detecting fetuin-B in actual clinical samples, thereby demonstrating its potential for preliminary CAD screening and diagnosis.

Prevalence of hepatitis B virus infection among pregnant women in the mountainous regions of southern China: A retrospective single-center study.

BACKGROUND: Hepatitis B virus (HBV) infection remains a major public health issue worldwide. Moreover, its prevalence varies significantly in different geographic areas of China. The current study aimed to assess the prevalence of HBV infection among Hakka pregnant women in Meizhou, a remote mountainous region in southern China. METHODS: This research was performed between January 2015 and December 2020. In total, 16,727 pregnant women receiving antenatal care at Meizhou People's Hospital were included in the analysis. All pregnant women were screened for serum HBV markers. RESULTS: The prevalence rates of hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody positivity among the participants were 11.74% (n = 1964) and 48.00% (n = 8029), respectively. The overall prevalence rates of susceptibility to infection, HBV immunity, previous/occult infection, inactive HBsAg carrier, and active infection were 36.16%, 33.61%, 16.94%, 8.11%, and 2.30%, respectively. According to age distribution, the prevalence rate of HBsAg positivity elevated concomitantly with increasing age (p < 0.001). From 2015 to 2020, the prevalence rate of HBsAg positivity decreased from 14.50% to 8.19% and that of hepatitis B pre-core antigen positivity from 4.42% to 2.31%. In addition, pregnant women with HBsAg-positive status were more likely to present with gestational diabetes, thrombocytopenia, and anemia than those with HBsAg-negative status. CONCLUSION: The HBV infection rate remains high among pregnant women in the indigenous Hakka population in southern China. To prevent vertical transmission, cautious surveillance of maternal HBV infection status should be considered in Hakka pregnant women in Meizhou.

CircRNAs open a new era in the study of cardiovascular disease (Review).

Circular RNAs (circRNAs) are non­coding RNAs that are found in the cytoplasm or stored in the exosomes, where they are not affected by RNA exonucleases. CircRNAs are widely expressed in mammalian tissues and cells. A number of studies have suggested that circRNAs are associated with the physiology and pathology of cardiovascular diseases (CVDs). Therefore, circRNAs have been considered as effector molecules and biomarkers in the cardiovascular system. The present review article summarizes the biological origin and roles of circRNAs as well as the available databases and research methods for their identification. Furthermore, it describes their regulatory mechanisms in cardiovascular physiology and pathology, including the regulation of atherosclerosis, immunity, cell proliferation, apoptosis and autophagy. In addition, the current review discusses the unresolved problems in circRNA research and the application of circRNAs in the treatment of CVDs. Finally, the CVD­associated circRNAs are also reviewed.

Clinical evaluation of the T-SPOT.TB test for detection of tuberculosis infection in northeastern Guangdong Province, China.

OBJECTIVE: We evaluated clinical performance of the T-SPOT.TB test for detecting tuberculosis (TB) infection in Meizhou, China. METHODS: We enrolled 2,868 patients who underwent T-SPOT.TB, smear, and TB-DNA at the same time. The tests' sensitivity and specificity were evaluated and compared in different groups, and in pulmonary TB (PTB) and extrapulmonary TB (EPTB) subgroups. Receiver operator characteristic (ROC) curve analysis was used to evaluate T-SPOT.TB's diagnostic value and determine its cutoff value. RESULTS: T-SPOT.TB, TB-DNA, and sputum smear sensitivity was 61.44%, 37.12%, and 14.02%; and specificity was 76.49%, 99.20% and 99.60%, respectively. The T-SPOT.TB positive rate was higher in the PTB and EPTB subgroups than in patients with other pulmonary diseases (61.38% and 61.76% vs. 23.34%). The T-SPOT.TB test had better diagnostic accuracy and sensitivity when the positive cutoff value of marker ESAT-6 was 2.5 [area under ROC curve = 0.701, 95%CI 0.687-0.715] and marker CFP-10 was 6.5 [area under ROC curve = 0.669, 95%CI 0.655-0.683]. CONCLUSION: T-SPOT.TB sensitivity was higher than that of TB-DNA or sputum smear, but the specificity was lower. T-SPOT.TB had moderate sensitivity and specificity for diagnosing TB. T-SPOT.TB's new positive cutoff value may be clinically valuable according to ROC analysis.

The Serum Tumor Markers in Combination for Clinical Diagnosis of Lung Cancer.

BACKGROUND: To explore the clinical value of combined detection of serum tumor markers in lung cancer, including carcinoembryonic antigen (CEA), carbohydrate antigen 15-3 (CA15-3), cytokeratin 19 fragment (CYFRA 21-1), neuron specific enolase (NSE), and squamous carcinoma antigen (SCCA). METHODS: The expression levels were compared among groups, and the combined effects of these tumor markers in the diagnosis of lung cancer were analyzed. In addition, EGFR gene mutations were detected in some patients with NSCLC. RESULTS: There were 776 patients (age 59.78 ± 10.39 years) with lung cancer and 794 controls (age 58.26 ± 15.73 years) included in our study. In this study, tumor markers were detected in lung cancer patients and controls. Individual sensitivity of the tumor markers sorted in descending order were CEA > CYFRA21-1 > CA15-3 > NSE, and the specificities were NSE > CYFRA21-1 > CEA > CA15-3. The combination of CEA + CA15-3 + CYFRA21-1 + NSE ranked the highest in the sensitivity index (75.00%) and specificity index (98.61%) in lung cancer. In adenocarcinoma, the area under the ROC curve (AUROC) of CEA (0.665) and CYFRA21-1 (0.631) were higher than those of CA15-3 (0.559) and NSE (0.507). In squamous carcinoma, the AUC of CYFRA21-1 (0.722) and SCC (0.628) were higher than those of CEA (0.579), CA15-3 (0.524), and NSE (0.552). In small cell carcinoma, the AUC of NSE (0.654) was higher than those of CEA (0.616), CYFRA21-1 (0.555), and CA15-3 (0.482). CONCLUSIONS: These serum tumor markers are valuable indicators in the clinical use. The combination of tumor markers can be used as a method to improve the effectiveness of clinical diagnosis for lung cancer.

T cell co-stimulator inducible co-stimulatory (ICOS) exerts potential anti-atherosclerotic roles through downregulation of vascular smooth muscle phagocytosis and proliferation.

BACKGROUND: Atherosclerosis (AS) is a chronic inflammatory disease. The role of the immune system in the etiology of the disease, particularly T cells, has been widely studied and is well established. T cell activation directly regulates co-signaling molecules present in immune synapses. Targeting one or several of these co-signaling molecules can inhibit T cell-mediated inflammation and delay or reduce AS. In recent years, this strategy has increasingly become a research focus. As such, we explored the role and therapeutic potential of the T cell co-stimulatory molecule inducible co-stimulatory (ICOS) in AS. METHODS: We compared the expression of ICOS in early AS lesions occurring in ApoE-deficient (ApoE-KO) rats fed a fat-diet and wild type (WT) rats fed the same diet. Eight-week old ApoE-KO and WT rats [ApoE-KO(0) and WT(0)] were fed a high-fat diet for 16 weeks [ApoE-KO(16) and WT(16)]. ICOS expression in aortic tissues was analyzed by quantitative real-time PCR, western blot, and confocal microscopy. The effect of ICOS overexpression in a transfected human T cell line on the phagocytosis and proliferation of co-cultured human aortic smooth muscle cells (HASMCs) was studied in vitro. RESULTS: Compared with WT(0), ApoE-KO(0), and WT(16) rats, ICOS expression in ApoE-KO(16) rats was significantly down-regulated both at the mRNA and protein levels. In vitro experiments indicated that ICOS overexpression reduces phagocytosis and proliferation by HASMCs, and may therefore produce an anti-atherosclerotic effect. CONCLUSIONS: The immune synaptic co-signaling molecule ICOS has an anti-atherosclerotic effect through inhibition of HASMC phagocytosis and proliferation, and can be used to delay plaque formation during the early stages of AS.

Circulating microRNA expression profiling and bioinformatics analysis of patients with coronary artery disease by RNA sequencing.

BACKGROUND: MicroRNAs play a vital role in coronary artery disease. Abnormal expression of microRNAs has been found to be associated with the occurrence of CAD. METHODS: We identified significantly differentially expressed microRNAs in plasma between 40 patients with CAD and 10 controls with NCA using RNA sequencing. The differentially expressed microRNAs were analyzed for Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. RESULTS: Fifty cDNA libraries were constructed and sequenced, and a total of 1871.82 M raw reads were obtained, and 2135 microRNAs were found. Compared to the expressed microRNAs of NCA controls, 159 microRNAs were differentially expressed in CAD patients, including 119 upregulated microRNAs and 40 downregulated microRNAs. The top 10 upregulated miRNAs were miR-144-3p, miR-34a-5p, miR-15b-3p, miR-22-3p, miR-29b-3p, miR-1270, miR-6891-5p, miR-106a-5p, miR-15b-5p, and hsa-miR-499b-3p. The top ten downregulated miRNAs were miR-4437, miR-6842-3p, miR-4664-3p, miR-671-3p, miR-219a-1-3p, miR-7848-3p, miR-664a-3p, miR-1284, miR-361-3p, and miR-6780a-5p. The target genes of differentially expressed microRNAs were related to many basic biological terms, such as biological process, cellular component, and molecular function. According to the KEGG pathway analysis, the most enriched pathways of the differentially expressed microRNAs were endocytosis, focal adhesion, axon guidance, and so on. Furthermore, six upregulated and two downregulated microRNAs were detected by qRT-PCR (Quantitative Real-time PCR) and ROC analysis for diagnosing CAD. CONCLUSION: The results suggest that the expression levels of some microRNAs may play a vital role in the physiological and pathological course of CAD. Our study may provide useful information for the diagnosis and treatment of CAD.

Analysis of genotype distribution of thalassemia and G6PD deficiency among Hakka population in Meizhou city of Guangdong Province.

OBJECTIVE: The aim of the study was to explore genotype distribution thalassemia and G6PD deficiency in Meizhou city, China. METHODS: A total of 16 158 individuals were involved in thalassemia genetic testing. A total of 605 subjects were screened for common Chinese G6PD mutations by gene chip analysis. Genotypes and allele frequencies were analyzed. RESULTS: A total of 5463 cases carried thalassemia mutations were identified, including 3585 cases, 1701 cases, and 177 cases with α-, ß-, and α + ß-thalassemia mutations, respectively. --SEA (65.12%), -α3.7 (19.05%), and -α4.2 (8.05%) deletion were the main mutations of α-thalassemia, while IVS-II-654(C â†’ T) (40.39%), CD41-42(-TCTT) (32.72%), -28(A â†’ G) (10.11%), and CD17(A â†’ T) (9.32%) mutations were the principal mutations of ß-thalassemia in Meizhou. There were significant differences in allele frequencies in some counties. Genetic testing for G6PD deficiency, six mutation sites, and one polymorphism were detected in our study. A total of 198 alleles with the mutation were detected among 805 alleles (24.6%). G6PD Canton (c.1376 G â†’ T) (45.96%), G6PD Kaiping (c.1388 G â†’ A) (39.39%), and G6PD Gaohe (c.95 A â†’ G) (9.09%) account for 94.44% mutations, followed by G6PD Chinese-5 (c.1024 C â†’ T) (4.04%), G6PD Viangchan (c.871G â†’ A) (1.01%), and G6PD Maewo (c.1360 C â†’ T) (0.51%). There were some differences of the distribution of G6PD mutations among eight counties in Meizhou. CONCLUSIONS: The --SEA , -α3.7 , and -α4.2 deletion were the main mutations of α-thalassemia, while IVS-II-654(C â†’ T), CD41-42(-TCTT), -28(A â†’ G), and CD17(A â†’ T) mutations were the principal mutations of ß-thalassemia in Meizhou. G6PD c.1376 G â†’ T, c.1388 G â†’ A, and c.95 A â†’ G were the main mutations of G6PD deficiency. There were some differences of the distribution of thalassemia and G6PD mutations among eight counties in Meizhou.

Differentially expressed microRNAs in exosomes of patients with breast cancer revealed by next­generation sequencing.

MicroRNAs (miRNAs/miRs) in exosomes play crucial roles in the onset, progression and metastasis of cancer by regulating the stability of target mRNAs or by inhibiting translation. In the present study, differentially expressed miRNAs were identified in exosomes of 27 breast cancer patients and 3 healthy controls using RNA sequencing. The differentially expressed microRNAs were selected by bioinformatic analysis. Subjects were followed up for 2 years and exosomal miRNA profiles were compared between patients with and without recurrence of breast cancer. A total of 30 complementary DNA libraries were constructed and sequenced and 1,835 miRNAs were detected. There were no significant differences in the expression of miRNAs between the basal­like, human epidermal growth factor receptor­2+, luminal A, luminal B and healthy control (HC) groups. A total of 54 differentially expressed miRNAs were identified in triple­negative breast cancer (TNBC) patients vs. HCs, including 20 upregulated and 34 downregulated miRNAs. The results of the reverse transcription­quantitative PCR were consistent with this. Receiver operating characteristic curve analyses indicated that miR­150­5p [area under the curve (AUC)=0.705, upregulated], miR­576­3p (AUC=0.691, upregulated), miR­4665­5p (AUC=0.681, upregulated) were able to distinguish breast cancer patients with recurrence from those without recurrence. In conclusion, the present results indicated differences in miRNA expression profiles between patients with TNBC and healthy controls. Certain exosomal miRNAs were indicated to have promising predictive value as biomarkers for distinguishing breast cancer with recurrence from non­recurrence, which may be utilized for preventive strategies.

Frequency of BRCA1 and BRCA2 Mutations in Individuals with Breast and Ovarian Cancer in a Chinese Hakka Population Using Next-Generation Sequencing.

BACKGROUND: It is necessary to investigate the frequency of BRCA1 and BRCA2 mutations in Hakka populations due to the variations in breast cancer epidemiology and genetics. METHODS: 359 breast cancer patients and 66 ovarian cancer patients were included in this retrospective clinical study. Mutations of BRCA1 and BRCA2 were detected in blood samples by semiconductor sequencing. RESULTS: The sensitivity of tumor markers including CEA, CA15-3, CA12-5, and CA199 for screening breast cancer was 16.44, 15.11, 8.44, and 7.56%, the combination of these 4 tumor markers reached the highest sensitivity index (31.11%). For ovarian cancer, the tumor markers were CA12-5 (54.05%), HE-4 (54.05%), CA72-4 (51.35%), and CEA (2.70%) in order of decreasing sensitivity. Moreover, the combination of these 4 tumor markers has the best sensitivity (75.68%) for screening ovarian cancer. In breast cancer patients, we found 5 (1.39%) patients with mutations in BRCA1, 13 (3.62%) mutations in BRCA2, and the total carrier rate is 5.01% (18/359). For ovarian cancer patients, the corresponding results were 3 (4.54%) mutations, 2 (3.03%) mutations, and 7.58% (5/66), respectively. The proportion of BRCA mutations was 5.41% (23/425) in breast and ovarian cancer patients of a Hakka population. The pathogenic, likely pathogenic, and benign mutations, and mutations of uncertain significance in this study mainly occurred in exon 14 of the BRCA1 gene, and exon 10 and exon 11 of the BRCA2 gene. CONCLUSIONS: Understanding the spectrum and frequency of BRCA1 and BRCA2 mutations in a Hakka population will assist in the prevention and control of hereditary breast and ovarian cancers in this population.

Development and validation of a novel diagnostic nomogram model based on tumor markers for assessing cancer risk of pulmonary lesions: A multicenter study in Chinese population.

PURPOSE: This study aimed to build a valid diagnostic nomogram for assessing the cancer risk of the pulmonary lesions identified by chest CT. PATIENTS AND METHODS: A total of 691 patients with pulmonary lesions were recruited from three centers in China. The cut-off value for each tumor marker was confirmed by minimum P value method with 1000 bootstrap replications. The nomogram was based on the predictive factors identified by univariate and multivariate analysis. The predictive performance of the nomogram was measured by concordance index and calibrated with 1000 bootstrap samples to decrease the overfit bias. We also evaluated the net benefit of the nomogram via decision curve analysis. Finally, the nomogram was validated externally using a separate cohort of 305 patients enrolled from two additional institutions. RESULTS: The cut-off for CEA, SCC, CYFRA21-1, pro-GRP, and HE4 was 4.8 ng/mL, 1.66 ng/mL, 1.83 ng/mL, 56.55 pg/mL, and 63.24Lpmol/L, respectively. Multivariate logistic regression model (LRM) identified tumor size, CEA, SCC, CYFRA21-1, pro-GRP, and HE4 as independent risk factors for lung cancer. The nomogram based on LRM coefficients showed concordance index of 0.901 (95% CI: 0.842-0.960; P < 0.001) for lung cancer in the training set and 0.713 (95% CI: 0.599-0.827; P < 0.001) in the validation set. Decision curve analysis reported a net benefit of 87.6% at 80% threshold probability superior to the baseline model. CONCLUSION: Our diagnostic nomogram provides a useful tool for assessing the cancer risk of pulmonary lesions identified in CT screening test.