Markers Neuron-specific Enolase Research Articles

Immunoaffinity Intact Top-Down Mass Spectrometry for Quantification of Neuron-Specific Enolase Gamma, a Low-Abundance Protein Biomarker.

Quantification of intact proteins in serum by liquid chromatography high-resolution mass spectrometry (HRMS) may be a useful alternative to bottom-up LC-MS or conventional ligand binding assays, due to reduced assay complexity and by providing additional information, such as isoform differentiation or detection of post-translational modifications. The 47.2 kDa lung cancer tumor marker neuron-specific enolase γ (NSEγ) was quantified in a clinically relevant concentration range of 6.25 to 100 ng/mL in NSE-depleted human serum using magnetic bead immunoprecipitation coupled to LC-high-resolution quadrupole-time-of-flight MS. The novelty of the described approach is in the combined setup of immunoaffinity extraction and the use of a full-length NSEγ calibrator and labeled NSEγ internal standard (IS) to reliably quantify the post-translationally acetylated form of this protein tumor marker in a top-down proteomics workflow. Isolation parameters and quantification using deconvolution and reconstructed extracted ion chromatograms were evaluated, and the development of a suitable liquid chromatography method was demonstrated. Various validation parameters were determined using both quantification methods, both showing acceptable performance. Additionally, deconvolution-based quantification enabled an accurate mass determination. The developed method was compared to a commercially available ECLIA and showed good correlation in sera of patients suspected of lung cancer. This assay may form the starting point for the development of a reference method for the standardization of immunoassays.

Elevated serum miR-142-5p correlates with ischemic lesions and both NSE and S100β in ischemic stroke patients.

This study aims to evaluate the correlation between miRNAs and known nerve injury markers neuron-specific enolase (NSE) and S100β in ischemic stroke (IS) patients, exploring its efficacy. We retrospectively analyzed 86 IS patients and 32 healthy controls. Clinical and neurological examinations were performed in the admitted patients and the severity of neurological deficits was assessed by National Institutes of Health Stroke Scale (NIHSS). Plasma extraction and serum isolation were performed on all subjects before and 2 weeks after admission. miR-142-5p in serum, and NSE and S100β contents were measured by RT-qPCR and ELISA. Ischemic lesions were more severe in IS patients, and NSE and S100β were abnormally elevated. miR-142-5p in the serum of IS patients was 2.85 times higher. After 2 weeks of treatment, serum miR-142-5, NSE, and S100β decreased. Patients' serum levels of miR-142-5p were 57.5% lower. Serum miR-142-5, NSE, and S100β were lower in patients with disease improvement than in patients with poor recovery. Additionally, miR-142-5 was positively correlated with NSE (P < 0.0001) and S100β (P = 0.0147), and also with the NIHSS score (P = 0.0004). miR-142-5p, NSE, and S100β in peripheral blood (PB) of IS patients are elevated, and miR-142-5p is positively correlated with NSE and S100β.

Characterisation of plasma proteins and circulating microRNAs in out-of-hospital cardiac arrest patients - approaches for predicting outcome

Abstract Rationale: Early prognostication after out-of-hospital cardiac arrest (OHCA) is an unmet clinical need. There is limited data on serial release kinetics of cardiac, brain and systemic biomarkers such as Neuron Specific Enolase (NSE) and S-100B. miRNAs are sensitive biomarker candidates in acute myocardial infarction (MI). Objective The objective was to characterise circulating biomolecules in the early release kinetics after OHCA, compared with a STEMI control group and assessed with clinical outcome. Methods and Results Serial blood samples of patients with OHCA with STEMI (n=20) and STEMI control (n=20) were collected at baseline and at 6, 12, 24 and 48h after admission. Untargeted plasma proteomics were performed using data-independent acquisition–mass spectrometry (DIA–MS). Candidate proteins and miRNAs were quantified. Proteomics returned n=156 proteins with significantly different kinetics between OHCA and STEMI. Six clusters, depicting distinct biological pathways, with characteristic differences between OHCA and STEMI were identified. The most distinct and significant differences between OHCA and STEMI patients were seen for acute phase reactants LBP, AACT, CRP and the lipid metabolite A2GL as well as endothelial and platelet miR-126 and cardiomyocyte-apoptosis/ischemia-reperfusion related miR-320a. Cardiogenic shock was best characterised by differential kinetics of coagulation and complement related proteins FA9, HRG, FHR1 and inflammation related miR-146. The primary endpoint was poor neurological recovery at 30-days. Individuals with good outcome showed distinct differences in adaptive immune response, complement and coagulation cascades and hemostasis. Circulating plasma levels of hypoxia-associated miR-101, miR-210 and S-100B protein were significantly elevated at baseline in OHCA patients. These molecules further showed higher baseline values in patients with poor outcome. For current prognosis marker NSE, this difference only develops after 48h. Cardiac/muscle miRNAs followed an early release-pattern similar to high-sensitivity cardiac troponin-T (hs-cTnT). Machine learning models were used to calculate the AUC for the prediction of poor neurological outcome at baseline. This was highest for hypoxia-induced miR-210 (AUC: 0.854), which performed significantly better than the protein biomarkers NSE (AUC: 0.646) and S-100B (AUC:0.656). Conclusions Untargeted mass spectrometry reveals distinct biological pathways and candidate biomolecules affected in patients suffering from OHCA compared with STEMI controls, in patients developing cardiogenic shock and in those with adverse versus good neurological outcome. Platelet-, inflammation and hypoxia associated miRNAs are associated with OHCA, cardiogenic shock and prognostication of adverse neurological outcome. Pronounced differences in early release kinetics of RNA and protein biomarkers may improve identification of OHCA patients at risk of poor outcome.

Effects of Parecoxib Sodium on Early Cognitive Impairment and Inflammation Levels in Burned Rats.

To study the effect of parecoxib sodium in alleviating inflammation in burned rats and restoring cognitive function in burned rats. A total of 30 specific pathogen free grade Sprague-Dawley rats were randomly divided into 6 groups: (1) blank control group (group C), (2) Sham surgery group (group Sham), (3) second-degree burn model (group B), (4) low-dose (1 mg/kg/d) parecoxib sodium (group L + B), (5) medium-dose (10 mg/kg/d) parecoxib sodium (group M + B), and (6) high-dose (20 mg/kg/d) parecoxib sodium (group H + B). ELISA measures inflammatory factors interleukin (IL)-2, IL-6, tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ), and cognitive function factors neuron-specific enolase (NSE), cortisol, and S-100β. Combined with water maze and dark-avoidance experiments to further verify the recovery of cognitive function in rats. The contents of IL-2, TNF-α, and IL-6 in group M + B were significantly lower than those in group Sham (P < .05), and the content of IFN-γ was significantly lower than that in group Sham (P < .05). The cognitive markers NSE, S-100β, and cortisol levels in group M + B were significantly higher than those in group Sham at 2 h, 1 d, 5 d, and 10 d after operation (P < .05). In the group M + B dark-avoidance experiment, the number of probes and errors was not significantly different than those in group Sham and group C (P > .05), and the number of times group M + B found a platform in the water maze experiment and crossed the platform was second only to group B and group C. Parecoxib sodium can effectively reduce inflammation in burn rats and promote cognitive recovery in burn rats, and the optimal dose of parecoxib sodium for burn rats is 10 mg/kg.

Efficient CdIn2S4/MgIn2S4 heterojunction for ultrasensitive detection of lung cancer marker neuron-specific enolase

In this work, a photoelectrochemical (PEC) immunosensor was constructed for the ultrasensitive detection of lung cancer marker neuron-specific enolase (NSE) based on a microflower-like heterojunction of cadmium indium sulfide and magnesium indium sulfide (CdIn2S4/MgIn2S4, CMIS) as photoactive material. Specifically, the well-matched energy level structure and narrow energy level gradients between CdIn2S4 and MgIn2S4 could accelerate the separation of electron-hole (e−-h+) pairs in the CMIS heterojunction to enhance the photocurrent of CMIS, which was increased 5.5 and 80 times compared with that of single CdIn2S4 and MgIn2S4, respectively. Meanwhile, using CMIS as photoactive material, increasing the biocompatibility by dropping Pt NPs on the surface of CMIS to immobilize the antibody through Pt–N bond. Fe3O4-Ab2, acting as the quencher, competitively consumes electron donors and absorbs light, leading to photocurrent quenching. With the increasing of quencher, the photocurrent decreased. Hence, the developed “signal-off” PEC immunosensor realized the trace detection of NSE within the range from 1.0 fg/mL to 10 ng/mL with a low detection limit of 0.34 fg/mL. This strategy provided a new perspective for establishing ternary metal sulfide heterojunction to construct PEC immunosensor for sensitive detection of disease biomarkers.

Immunoaffinity Intact-Mass Spectrometry for the Detection of Endogenous Concentrations of the Acetylated Protein Tumor Biomarker Neuron Specific Enolase.

Intact-mass spectrometry has huge potential for clinical application, as it enables both quantitative and qualitative analysis of intact proteins and possibly unlocks additional pathophysiological information via, e.g., detection of specific post-translational modifications (PTMs). Such valuable and clinically useful selectivity is typically lost during conventional bottom-up mass spectrometry. We demonstrate an innovative immunoprecipitation protein enrichment assay coupled to ultrahigh performance liquid chromatography quadrupole time-of-flight high resolution mass spectrometry (UPLC-QToF-HRMS) for the fast and simple identification of the protein tumor marker Neuron Specific Enolase Gamma (NSEγ) at low endogenous concentrations in human serum. Additionally, using the combination of immunoaffinity purification with intact mass spectrometry, the presence of NSEγ in an acetylated form in human serum was detected. This highlights the unique potential of immunoaffinity intact mass spectrometry in clinical diagnostics.

Markers of brain injury in patients with aneurysmal subarachnoid hemorrhage

Aim To investigate the correlation of serum changes and markers of brain injury (BI) in cerebrospinal fluid (CSF) with postoperative cognitive dysfunction (POCD) in patients with cerebral aneurysmal subarachnoid haemorrhage (aSAH). Methods 120 patients diagnosed with aSAH were included. 3 months after surgery, these patients were divided into a normal cognition group and a cognitive dysfunction (CD) group relying on the Montreal Cognitive Assessment (MoCA) Scale. Results The correlations were analysed between the serological changes and the levels of BI markers, such as neurofilament-light (NF-L) protein, Ubisquitin C-terminal hydrolase L1(UCH-L1), Glial Fibrillary Acidic Protein (GFAP), and neuron specific enolase (NSE) in patients after surgery. Hunt-Hess grading standard was employed to determine the severity of aSAH in patients. The mean values of NF-L, UCH-L1, GFAP, and NSE were (8.2 ± 4.3) pg/mL, (0.7 ± 0.3) ng/mL, (2.2 ± 0.4) ng/mL, and (48.5 ± 10.9) ng/mL in patients with severe aSAH, which were remarkably higher than those in patients with mild aSAH [(3.5 ± 0.7) pg/mL, (0.5 ± 0.2) ng/mL, (1.3 ± 0.7) ng/mL, (30.7 ± 8.2) ng/mL]. The sensitivity, specificity, and accuracy of the combined prediction of four detections for POCD were 90.80%, 84.20%, and 82.80%, respectively, which were greatly higher than those of four independent predictions (p < 0.05). The combined prediction effect of the four items, with the area under the curve (AUC) of 0.938 and the 95% confidence interval (CI) of 0.851-0.926. Conclusions BI markers NF-L, UCH-L1, GFAP, and NSE could be utilized as predictors of POCD in patients with aSAH, deserving a reference value.

Moxibustion preconditioning reduces inflammatory response in rats with cerebral ischemia-reperfusion injury by regulating PI3K / AKT / mTOR signaling pathway.

To observe the effect of moxibustion preconditioning on inflammatory response in rats with cerebral ischemia reperfusion injury (CIRI), so as to explore its mechanisms underlying improving CIRI. Seventy-five male SD rats were randomly divided into sham operation, model, moxibustion preconditioning 3 days (Moxi 1), moxibustion preconditioning 5 days (Moxi 2) and moxibustion preconditioning 7 days (Moxi 3) groups, with 15 rats in each group. Moxibustion was applied at "Baihui"(GV20), "Dazhui"(GV14) and "Zusanli"(ST36) for 20 min once a day, totally for 3, 5 or 7 days. Thirty minutes after the last moxibustion treatment, the CIRI model was established by occlusion of the middle cerebral artery. The neurological deficit score was assessed by using Longa's method. The infarct size of the brain assessed after staining with 2% triphenyltetrazolium chloride (TTC). The morphological changes of cortical neurons were observed by HE staining. The contents of inflammatory factors interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), S-100β protein (S-100β) and neuron-specific enolase (NSE) were detected by ELISA. The expression of phosphatidylinositol-3-kinase (PI3K), p-PI3K, protein kinase B (AKT) and mammalian target of rapamycin (mTOR) proteins in the ischemic cortex tissues were detected by immunohistochemistry and Western blot. Compared with the sham operation group, the neurological function score and the percentage of cerebral ischemic volume were increased (P<0.01). The contents of serum IL-1β, TNF-α, S-100β and NSE were significantly increased (P<0.01), while the protein expressions of PI3K, p-PI3K, AKT and mTOR in the cerebral cortex were significantly decreased (P<0.01) in the model group. Compared with the model group, the neurological function score and the percentage of cerebral ischemic volume were significantly decreased (P<0.01). The contents of serum IL-1β, TNF-α, S-100β and NSE were significantly decreased (P<0.01), and the expressions of PI3K, p-PI3K, AKT and mTOR proteins in the cerebral cortex were significantly increased (P<0.01) in three moxibustion groups. Compared with the Moxi 1 and Moxi 2 groups, the above indicators were significantly improved in rats of the Moxi 3 group (P<0.01, P<0.05). Moxibustion preconditioning can significantly improve the neurological function of rats after ischemia-reperfusion, inhibit serum inflammatory factors IL-1 β and TNF-α, inhibit brain tissue injury markers S-100β and NSE, which may be related to the activation of PI3K/AKT/mTOR signaling pathway. The protective effect of moxibustion preconditioning for 7 days on CIRI was better than that of 5 days and 3 days.

Prognostic value of tumor markers ProGRP, NSE and CYFRA 21-1 in patients with small cell lung cancer and chemotherapy-induced remission.

Despite successful response to first line therapy, patients with small-cell lung cancer (SCLC) often suffer from early relapses and disease progression. To investigate the relevance of serum tumor markers for estimation of prognosis at several time points during the course of disease. In a prospective, single-center study, serial assessments of progastrin-releasing peptide (ProGRP), neuron-specific enolase (NSE), cytokeratin-19 fragments (CYFRA 21-1) and carcino-embryogenic antigen (CEA) were performed during and after chemotherapy in 232 SCLC patients, and correlated with therapy response and overall survival (OS). ProGRP, NSE and CYFRA 21-1 levels decreased quickly after the first chemotherapy cycle and correlated well with the radiological response. Either as single markers or in combination they provided valuable prognostic information regarding OS at all timepoints investigated: prior to first-line therapy, after two treatment cycles in patients with successful response to first-line therapy, and prior to the start of second-line therapy. Furthermore, they were useful for continuous monitoring during and after therapy and often indicated progressive disease several months ahead of radiological changes. The results indicate the great potential of ProGRP, NSE and CYFRA 21-1 for estimating prognosis and monitoring of SCLC patients throughout the course of the disease.

Tumor protein D52 (isoform 3) induces NF-κB – STAT3 mediated EMT driving neuroendocrine differentiation of prostate cancer cells

In prostate cancer (PCa) patients, a proto-oncogene Tumor protein D52 (TPD52) is overexpressed, and it is involved in different cellular functions. In this study, we report that TPD52 expression is positively associated with the emergence of neuroendocrine PCa (NEPC). With overexpression of TPD52 in LNCaP cells, we found neuroendocrine differentiation (NED) of cells in in-vitro and distinct NED features confirmed by NE markers neuron-specific enolase (NSE) and chromogranin A (CHR-A). Further, we investigated the molecular mechanisms involved in TPD52 mediated NED of PCa cells. We found that TPD52 activates the NF- κB – STAT3 axis for the induction of NED in LNCaP cells. Indeed, inhibition of NF-κB – STAT3 attenuated the progression of NED in TPD52 positive LNCaP cells. Importantly, silencing of TPD52 expression or inhibition of NF-κB – STAT3 activity in a neuroendocrine cell line NCI-H660 showed a marked decrease in the expression of NSE and CHR-A, confirming the reversal of the NE properties. Notably, TPD52 overexpression in LNCaP cells induced expression of N-cadherin, Vimentin, ZEB1, and Snail1 indicating that TPD52 positively regulates epithelial to mesenchymal transition (EMT) of PCa cells towards NED. Moreover, silencing of Snail1 in TPD52 positive cells blocked the progression of NED and, in NCI-H660 cells reversed NE properties as expected. Of the few requirements of TPD52, activation of NF-κB – STAT3 is essential for promoting EMT compelling NED of LNCaP cells. Collectively, these results reveal that TPD52 is associated with the progression of NEPC and emphasizes the need for therapeutic targeting of TPD52 in PCa.

Effects of Computer Virtual Screening and Treatment Based on Pharmacophore BRD4-Targeted Small-Molecule Inhibitors on NSE and CA19-9 Levels in Patients with Liver Cancer

Objective: Neuron-specific enolase (NSE) and carbohydrate antigen 19-9 (CA19-9) levels were compared before and after treatment with brd4-targeting small-molecule inhibitors based on pharmacophores and computer technology to investigate an efficient virtual screening method for bromodomain protein 4 (brd4)-targeted small-molecule inhibitors. The purpose of this research was to examine the efficacy of pharmacophore screening and BRD4 targeting by small-molecule inhibitors in the management of liver cancer. Methods: Subjects included 106 individuals with hepatocellular cancer who were hospitalised between July 2019 and December 2021. Using a receiver operating characteristic (ROC) curve, we assessed the clinical relevance and utility of pharmacophore-based virtual screening for brd4-targeted small-molecule inhibitors. The levels of the tumour markers NSE and CA19-9 were analysed for changes using computational methods. Results: The pharmacophore model was built using the HipHop technique, and its results were validated and optimised utilising computational technology analysis. Twenty-four small-molecule classes were covered by the pharmacophore, and the number of mismatched pharmacodynamic characteristics was 0. The first eight pharmacophores each had two clusters of hydrophobic atoms, a hydrogen bond receptor, and an aromatic ring at its core. There were no changes to the pharmacodynamic characteristics, although their locations varied. False-positive and true-positive rates are depicted on the horizontal and vertical axes, respectively, in the figure below; the other two pharmacophore groups contained a hydrophobic cluster, a hydrogen bond receptor, and the centre of an aromatic ring. The ROC curve was quite consistent, as indicated by an AUC value close to 1, which was between 0 and 1. For pharmacophores 07 and 06, the area under the curve (AUC) was 0.949 and 0.955, respectively, very near to 1. Both the hydrophobic cluster and the hydrogen bond receptor are required for a pharmacophore to exist. The examination of pharmacodynamic definitions revealed a positive correlation between database reliability and element type (P &lt;0.05). Compared to pre-healing levels, post-healing CA19-9 and NSE levels were statistically considerably reduced (P &lt; 0.05). Conclusions: The assessment approach was highly accurate and specific, and the pharmacophore-based virtual screening method and computer technology for brd4-targeting small-molecule inhibitors were of great benefit in the regulation of the cell cycle and the process of cell proliferation. Effective treatment modalities can be created to increase levels of tumour markers NSE and CA19-9, and pharmacophore and computational-based small-molecule inhibitors of brd4 can be employed as crucial virtual screening criteria for the evaluation of liver cancer.

Analysis of Diagnostic Value of CTC and CTDNA in Early Lung Cancer.

This paper is the exploration and analysis of the diagnostic value of circulating tumor cells (CTC) and circulating tumor deoxyribonucleic acid (ctDNA) in early non-small cell lung cancer (NSCLC).A total of 125 NSCLC patients that hospitalized from March 2019 to December 2020 were selected, of which 64 subjects with stage I-II were listed as the early-stage group, and 61 patients with stage III-IV were classified as the intermediate-advanced group; In addition, 47 patients with benign pulmonary nodules hospitalized within same period were selected as a group of benign pulmonary nodules, and 50 healthy subjects were enrolled as a control group. The levels of CTC and ctDNA, serum tumor markers carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), cytokeratin 21-1 (CYFRA21-1) and gastrin-releasing peptide precursor (pro-GRP) were detected. The diagnostic value of CTC and ctDNA in early NSCLC patients was analyzed by ROC curve. The area under the curve of CTC count for early-stage lung cancer was 0. 949 (P=0. 000, 95%CI: 0. 918-0. 980), and ctDNA was 0. 914 (P=0. 000, 95%CI: 0. 866~0. 963). There were significant differences in CTC count and ctDNA among subjects in each group (P<0. 05). CTC count and ctDNA of the early and intermediate-advanced group were remarkably higher than those of the control group and group of benign pulmonary nodules (P<0. 05), and CTC count and ctDNA of patients in the middle-late group were critically higher than those in the early-stage group (P<0. 05). There were significant differences in serum CEA, NSE, CYFRA21-1 and pro-GRP levels among all groups (P<0. 05). The results showed that CTC and ctDNA in NSCLC patients were significantly positively correlated with CEA, NSE, CYFRA21-1 and pro-GRP (P<0. 05). Peripheral blood CTC and ctDNA levels are significantly increased in NSCLC patients. They are positively correlated with serum tumor markers CEA, NSE, CYFRA21-1 and pro-GRP levels, and are related to tumor progression. CTC and ctDNA have high value in the clinical diagnosis of early NSCLC.

Performance Evaluation of Combined Detection of Serum CEA, CYFRA21-1, CA125, and NSE in Patients with Lung Cancer by Fluorescence Flow Cytometry

Objective: To investigate the effect of combined detection of serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1), cancer antigen 125 (CA125), and neuron-specific enolase (NSE) in patients with lung cancer by fluorescence flow cytometry. Methods: From August 2019 to July 2022, 200 patients with lung cancer diagnosed by pathology in our hospital were retrospectively analyzed. 2 mL venous blood was collected in a fasting state and centrifuged to separate the serum (containing human chorionic gonadotropin antibody [anti-hCG antibody], hepatitis B surface antibody [anti-HBs antibody], and CEA). Results: The sensitivities of CEA and CYFRA21-1 detected via enzyme-linked immunosorbent assay (ELISA) were 100%, and the detection limits were 0.5 ng/mL and 0.1 ng/mL, respectively; the sensitivities of CA125 and NSE detected via flow cytometry were 100%, and the detection limits were 10 U/mL and 2 ng/mL, respectively. Compared with ELISA, the sensitivities of CA125 and NSE detected via flow cytometry were higher. When the concentration of CEA was 10–40 ng/mL, the sensitivities of the three markers CYFRA21-1, CA125, and NSE showed no significant changes (P &gt; 0.05); when the concentration of CEA was 40–80 ng/mL, the sensitivity of CEA significantly decreased (P &lt; 0.01), but the sensitivities of the three markers CYFRA21-1, CA125, and NSE showed no significant changes (P &gt; 0.05); when the concentration of CEA was 80–200 ng/mL, the sensitivities of all four markers showed no significant changes (P &gt; 0.05). Conclusion: Compared with the double-antibody sandwich ELISA, fluorescence flow cytometry has certain advantages, including high sensitivity, good precision, short detection time, low sample usage, and low medical cost; thus, it is worthy of clinical promotion.

Clinical role of serum tumor markers SCC, NSE, CA 125, CA 19-9, and CYFRA 21-1 in patients with lung cancer.

The aim of the study was to assess the diagnostic value of tumor markers in discriminating between lung cancer and benign chest diseases (BCDs). There were 322 patients enrolled in this investigation including 180 cases of lung cancer and 142 cases of BCD. Serum neuron-specific enolase (NSE), cancer antigen 125, cancer antigen 19-9, squamous cell carcinoma-related antigen, and cytokeratin fragment19 (CYFRA 21-1) were compared between different populations, cancer stages, and before and after treatment. Logistic regression and receiver operating characteristic curves were used to evaluate the diagnostic markers. Both NSE and CYFRA 21-1 were significantly associated with lung cancer. The CYFRA 21-1 showed the best performance, as well as its combinations, for lung cancer diagnosis. It also showed significant change 6 months after radical surgery in lung cancer patients. The marker CYFRA 21-1 could be developed as an adjuvant marker for the early diagnosis of lung cancer and as a prognostic marker for lung cancer treatment.

Advantages and clinical significance of enhanced CT combined with BRAF V600E gene detection in the diagnosis of papillary thyroid carcinoma

The diagnosis of papillary thyroid carcinoma (PTC) remains the focus of clinical research. Studies have shown that the BRAFV600E gene mutation is closely related to the development of PTC. In this study, we used the advantages and clinical significance of enhanced CT combined with BRAFV600E gene testing in the diagnosis of PTC. A total of 170 PTC patients who received treatment in our hospital between January 2020 and December 2020 were collected and divided into group A (enhanced CT combined with BRAFV600E gene detection) and group B (fine needle aspiration cytology, FNAC) according to different diagnostic methods. We recorded the general information of the patients and measured the levels of serum tumor markers including carcinoembryonic antigen (CEA), cytokeratin fragment 19 (CYFRA21-1), neuron-specific enolase (NSE) and carbohydrate antigen 125 (CA125). After pathological diagnosis, the specificity, sensitivity, accuracy, positive predictive value, and negative predictive value of the two diagnostic methods were contrasted. The accuracy of the two methods and the pathological test in identifying benign lesions, malignant tumors, was also compared. All patients' serum tumor markers CEA, CYFRA21-1, NSE and CA125 levels were above normal. The detection rate of specificity, sensitivity, accuracy, positive predictive value and negative predictive value of enhanced CT combined with BRAF V600E gene detection were superior to those of FNAC. And the accuracy of enhanced CT combined with BRAF V600E gene detection, which was confirmed by corresponding pathological detection results, was significantly increased (P<0.05). Enhanced CT combined with BRAF V600E gene detection can improve the diagnostic accuracy of PTC, and its clinical indicators and safety are superior to FNAC.

Diagnostic value of serum tumor markers CEA, CYFRA21-1, SCCAg, NSE and ProGRP for lung cancers of different pathological types

To evaluate the diagnostic value of the serum tumor markers carcinoembryonic antigen (CEA), cytokeratin-19-fragment (CYFRA21-1), squamous cell carcinoma associated antigen (SCCAg), neuron-specificenolase (NSE) and pro-gastrin-releasing peptide (ProGRP) for lung cancers of different pathological types. This study was conducted among patients with established diagnoses of lung adenocarcinoma (LADC, n=137), lung squamous cell carcinoma (LSCC, n=82), small cell lung carcinoma (SCLC, n=59), and benign chest disease (BCD, n=102). The serum tumor markers were detected for all the patients for comparison of the positivity rates and their serum levels. ROC curve was used for analysis of the diagnostic efficacy of these tumor markers either alone or in different combinations. In patients with LADC, the positivity rate and serum level of CEA were significantly higher than those in the other groups (P < 0.05); the patients with LSCC had the highest positivity rate and serum level of SCCAg among the 4 groups (P < 0.05). The positivity rates and serum levels of ProGRP and NSE were significantly higher in SCLC group than in the other groups (P < 0.05). CYFRA21-1 showed the highest positivity rate and serum level in LADC group and LSCC group. With the patients with BCD as control, CEA showed a diagnostic sensitivity of 62.8% and a specificity of 93.1% for LADC, and the sensitivity and specificity of SCCAg for diagnosing LSCC were 64.6% and 91.2%, respectively. CYFRA21-1 had the highest diagnostic sensitivity for LADC and LSCC. The diagnostic sensitivity and specificity of ProGRP for SCLC were 83.1% and 98.0%, respectively. When combined, CYFRA21-1 and CEA showed a high sensitivity (78.8%) and specificity (86.3%) for diagnosing LADC with an AUC of 0.891; CYFRA21-1 and SCCAg had a high sensitivity (84.1%) and specificity (87.3%) for diagnosing LSCC with an AUC of 0.912. NSE combined with ProGRP was highly sensitive (88.1%) and specific (98.0%) for diagnosis of SCLC, with an AUC of 0.952. For lung cancers of different pathological types, the combination of all the 5 tumor markers showed no significant differences in the diagnostic power from a combined detection with any two of the markers (P>0.05). CEA, CYFRA21-1, SCCAg, NSE and ProGRP are all related to the pathological type of lung cancers and can be used in different combinations as useful diagnostic indicators for lung cancers.

Reprogramming of Rat Fibroblasts into Induced Neurons by Small-Molecule Compounds In Vitro and In Vivo.

Cell replacement is a promising approach for neurodegenerative disease treatment. Somatic cells such as fibroblasts can be induced to differentiate into neurons by specific transcription factors; however, the potential of viral vectors used for reprogramming to integrate into the genome raises concerns about the potential clinical applications of this approach. Here, we directly reprogrammed rat embryonic skin fibroblasts into induced neurons (iNs) via six small-molecule compounds (SMs) (VPA, CHIR99021, forskolin, Y-27632, Repsox, and P7C3-A20). iNs exhibit typical neuronal morphology, and immunofluorescence showed that more than 96% of the iNs expressed the early neuronal marker class III beta-tubulin (TUJ1) and that more than 91% of iNs expressed the mature neuronal marker neuron-specific enolase (NSE) after 10 days of reprogramming. Quantitative real-time polymerase chain reaction also showed that most iNs expressed the dopaminergic neuron marker tyrosine hydroxylase, the neural marker Nur correlation factor 1, the (γ-aminobutyric acid, GABA) GABAergic neuronal marker GABA, and the cholinergic neuron marker choline acetyltransferase. In addition, we found that cell proliferation decreased during reprogramming and that protein synthesis increased initially and then decreased. SMs were mixed with hydrogels, and the hydrogels were implanted subcutaneously into the backs of rats. After 7 days, the TUJ1 and NSE proteins were expressed in surrounding tissues, indicating that SMs caused reprogramming in vivo. In summary, rat skin fibroblasts can be efficiently reprogrammed into iNs by SMs in vitro and in vivo.

Brain damage markers neuron-specific enolase (NSE) and S100B in serum in children with Lyme neuroborreliosis—detection and evaluation as prognostic biomarkers for clinical outcome

Lyme borreliosis (LB) is the most common tick-borne infection in Europe, with Lyme neuroborreliosis (LNB) its second most frequent clinical manifestation. Prognostic factors for clinical outcomes in LNB have not been identified. Elevated serum levels of the brain damage markers neuron-specific enolase (NSE) and S100 calcium-binding protein B (S100B) have been associated with poor clinical outcomes in other disorders of the central nervous system. The aim of this study is to assess NSE and S100B in serum as prognostic biomarkers for clinical outcomes in paediatric LNB patients. Children evaluated for LNB (n = 121) in Sweden were prospectively included during 2010–2014, serum samples were collected on admission, and all children underwent a 2-month follow-up. Patients with pleocytosis and anti-Borrelia antibodies in cerebrospinal fluid (CSF) were classified as having LNB (n = 61). Controls were age- and gender-matched non-LNB patients (n = 60). NSE was elevated in 38/61 (62%) LNB patients and in 31/60 (52%) controls. S100B was elevated in 3/60 (5%) LNB patients and 0/59 (0%) controls. NSE and S100B concentrations did not differ significantly when comparing LNB patients with controls. No differences were found in the concentrations when comparing the clinical recovery of LNB patients at the 2-month follow-up. NSE was detectable in the majority of LNB patients and controls, whereas S100B was detectable in only a few LNB patients and no controls. NSE and S100B in serum cannot be recommended as prognostic biomarkers for clinical outcomes in children with LNB.

Biomarkers of brain injury after cardiac arrest; a statistical analysis plan from the TTM2 trial biobank investigators

BackgroundSeveral biochemical markers in blood correlate with the magnitude of brain injury and may be used to predict neurological outcome after cardiac arrest. We present a protocol for the evaluation of prognostic accuracy of brain injury markers after cardiac arrest. The aim is to define the best predictive marker and to establish clinically useful cut-off levels for routine implementation. MethodsProspective international multicenter trial within the Targeted Hypothermia versus Targeted Normothermia after Out-of-Hospital Cardiac Arrest (TTM2) trial in collaboration with Roche Diagnostics International AG. Samples were collected 0, 24, 48, and 72 hours after randomisation (serum) and 0 and 48 hours after randomisation (plasma), and pre-analytically processed at each site before storage in a central biobank. Routine markers neuron-specific enolase (NSE) and S100B, and neurofilament light, total-tau and glial fibrillary acidic protein will be batch analysed using novel Elecsys® electrochemiluminescence immunoassays on a Cobas e601 instrument. ResultsStatistical analysis will be reported according to the Standards for Reporting Diagnostic accuracy studies (STARD) and will include comparisons for prediction of good versus poor functional outcome at six months post-arrest, by modified Rankin Scale (0–3 vs. 4–6), using logistic regression models and receiver operating characteristics curves, evaluation of mortality at six months according to biomarker levels and establishment of cut-off values for prediction of poor neurological outcome at 95–100% specificities. ConclusionsThis prospective trial may establish a standard methodology and clinically appropriate cut-off levels for the optimal biomarker of brain injury which predicts poor neurological outcome after cardiac arrest.

Low Doses of Imidacloprid Induce Oxidative Stress and Neural Cell Disruption in Earthworm &lt;i&gt;Eisenia fetida&lt;/i&gt;

Imidacloprid is a widely used pesticide that belongs to the class of neonicotinoids. There is a piece of rising evidence that neonicotinoids exert cytotoxic effects in non-target organisms including vertebrate species such as mammals. Nevertheless, dose-limiting toxicity and molecular mechanisms of neonicotinoids' deleterious effects are still poorly understood. In accord to imidacloprid fate in the environment, the most of used pesticide is absorbed in the soil. Therefore, earthworms, which are prevailing soil organisms, could be considered as a target of neonicotinoids toxicity. The earthworm’s simple nervous system is a prospective model for neurotoxicological studies. We exposed earthworms to imidacloprid in a paper contact test with a doses range of 0.1‑0.4 µg/cm2 for 14 days. In the present work, we studied the imidacloprid effect on oxidative stress generation and neuronal marker neuron-specific enolase (NSE) expression. The exposure to imidacloprid induced a dose-dependent decrease in NSE. Both reactive oxygen species production and lipid peroxidation level were upregulated as well. Observed NSE decline suggests imidacloprid-caused disturbance in earthworm neuron cells. Obtained data have shown that relatively low doses of imidacloprid are potent to induce cytotoxicity in neurons. Furthermore, neurotoxicity could be recognized as one of an individual scenario of the general imidacloprid toxicity. Thus, presented results suggest the cytotoxicity of imidacloprid low doses in non-target organisms and hypothesize that NSE downregulation could be estimated as a biomarker of neonicotinoid cytotoxicity in a nervous system of non-insect species.