Study on the value of small dense low-density lipoprotein in predicting cardiovascular and cerebrovascular events in the high-risk stroke population.

BACKGROUND: Lipid management in people at high risk of stroke is an important measurement to prevent the occurrence of stroke. The study aims to investigate the association between sdLDL and cardiovascular and cerebrovascular events in high-risk stroke populations. METHODS: This was a prospective study. Screened from 15,933 individuals aged >40 years in April 2013 and followed up at 3rd, 6th, 12th, and 24th months, 823 participants met the screening criteria and were investigated for clinical data and biochemical parameters. RESULTS: A total of 286 subjects had varying degrees of carotid stenosis, and 18 subjects experienced cardiovascular and cerebrovascular events during the two-year follow-up period. There was no positive correlation between sdLDL and carotid stenosis. Carotid stenosis and extent of carotid stenosis involvement did not predict cardiovascular and cerebrovascular events in patients with high-risk stroke, while sdLDL did. The sdLDL level in the events group was significantly higher than those in the no event group (p = 0.002). In the events group, the risk of events in the fourth quartile of sdLDL was 10.136 times higher than in the first quartile (HR = 10.136, 95% CI: 1.298-79.180, p = 0.027). CONCLUSIONS: sdLDL was positively correlated with the incidence of cardiovascular and cerebrovascular events, which can predict the occurrence of an event and provide a scientific basis for early prevention.

Multi-function PtCo nanozymes/CdS nanocrystals@graphene oxide luminophores and K2S2O8/H2O2 coreactants-based dual amplified electrochemiluminescence immunosensor for ultrasensitive detection of anti-myeloperoxidase antibody.

BACKGROUND: Anti-myeloperoxidase antibody (anti-MPO) is an important biomarker for anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAVs). However, the complicated operation procedures and insufficient sensitivity of conventional anti-MPO detection methods limit their application in monitoring efficacy of AAVs in clinical diagnosis. Herein, a dual amplified electrochemiluminescence (ECL) immunosensor based on multi-function PtCo nanozymes/CdS nanocrystals@graphene oxide (PtCo/CdS@GO) luminophores and K2S2O8/H2O2 coreactants has been fabricated for ultrasensitive detection of anti-MPO. RESULTS: PtCo/CdS@GO luminophores as novel signal amplification labels and nanocarriers to load rabbit anti-mouse IgG were synthesized by co-doping with Pt and Co nanozymes simultaneously with several considerable advantages, including astonishing peroxidase-like catalytic activity, high-efficiency luminescence performance and superior stability in aqueous solutions. Meanwhile, upon the K2S2O8/H2O2 coreactants system, benefiting from the efficient peroxidase-like activity of the PtCo/CdS@GO toward H2O2, massive of transient reactive intermediates could react with K2S2O8, thus obtaining higher ECL emission. Therefore, the developed ECL immunosensor for anti-MPO detection displayed good analytical performance with good concentration linearity in the range of 0.02 to 1000 pg/mL and low detection limit down to 7.39 fg/mL. CONCLUSIONS: The introduction of multi-function PtCo/CdS@GO luminophores into the established ECL immunoassay not only was successfully applied for specific detection of anti-MPO in clinical serum samples, but also provided a completely new concept to design other high-performance luminophores. Meaningfully, the ECL immunoassay strategy held wide potential for biomarkers detection in clinical diagnosis.

Lp-PLA2 evaluates the severity of carotid artery stenosis and predicts the occurrence of cerebrovascular events in high stroke-risk populations.

BACKGROUND: Lipoprotein-associated phospholipase A2 (Lp-PLA2) is an independent risk factor for cardiovascular disease. However, relationship between carotid artery stenosis and cerebrovascular events in high stroke-risk populations is still unclear. METHODS: A total of 835 people at a high risk of stroke were screened from 15,933 people aged >40 years in April 2013 and followed at 3, 6, 12, and 24 months. Finally, 823 participants met the screening criteria, and the clinical data and biochemical parameters were investigated. RESULTS: Among the 823 participants, 286 had varying degrees of carotid artery stenosis and 18 had cerebrovascular events. The level of Lp-PLA2 in the carotid artery stenosis group was higher than that in the no stenosis group, and the level in the event group was higher than that in the no event group (p < 0.05). Spearman correlation analysis showed that Lp-PLA2 was positively correlated with the degree of carotid artery stenosis (r = 0.093, p = 0.07) and stenosis involvement (r = 0.094, p = 0.07). The correlation coefficient between Lp-PLA2 and lipoprotein was the highest on the levels of sdLDL (r = 0.555, p < 0.001), followed by non-HDL, LDL, TC, and TG. Cox multivariate regression analysis revealed that, compared with the first quantile of Lp-PLA2 level (Q1, low level), the risk of cerebrovascular events in the fourth quantile of Lp-PLA2 was 10.170 times that of the first quantile (OR = 10.170, 95% CI 1.302-79.448, p = 0.027). CONCLUSIONS: Lp-PLA2 levels can evaluate carotid artery stenosis and predict the occurrence of cerebrovascular events in high stroke-risk populations and provide scientific guidance for risk stratification management.

Establishment of the reference intervals of whole blood neutrophil phagocytosis by flow cytometry.

OBJECTIVE: To investigate the reference intervals (RIs) of the whole blood neutrophil phagocytosis by flow cytometry (FCM) and to study the application value of neutrophil phagocytosis in infectious diseases. METHODS: Pathogens (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923) cultured for 18-24 h were labeled by fluorescence probe carboxyfluorescein diacetate succinimidyl ester (CFDA-SE), and then incubated with whole blood at 37℃. The phagocytosis of pathogens by neutrophils was detected by flow cytometry, and a reference interval was established. RESULTS: In the healthy adults, the reference interval for the neutrophil phagocytosis to Escherichia coli was 46.91%-83.09% and to Staphylococcus aureus was 33.92%-69.48%. This method showed good reproducibility. Neutrophil phagocytosis was negatively correlated with the neutrophil count, neutrophil percentage, and neutrophil-to-lymphocyte ratio (NLR, p < 0.05). CONCLUSION: We have successfully established the RIs of neutrophil phagocytosis in whole blood in healthy adults by flow cytometry (FCM), which might be of important clinical value in the diagnosis, treatment, and prognosis of infectious diseases.

Establishment of a regression model of bone metabolism markers for the diagnosis of bone metastases in lung cancer.

BACKGROUND: The aim of this study was to establish a regression equation model of serum bone metabolism markers. We analyzed the diagnostic value of bone metastases in lung cancer and provided laboratory evidence for the early clinical treatment of bone metastases in lung cancer. METHODS: A total of 339 patients with non-metastatic lung cancer, patients with lung cancer with bone metastasis, and patients with benign lung disease who were treated in our hospital from July 2012 to October 2015 were included. A total of 103 patients with lung cancer in the non-metastatic group, 128 patients with lung cancer combined with bone metastasis group, and 108 patients with benign lung diseases who had nontumor and nonbone metabolism-related diseases were selected as the control group. Detection and analysis of type I collagen carboxyl terminal peptide ß-special sequence (ß-CTX), total type I procollagen amino terminal propeptide (TPINP), N-terminal-mid fragment of osteocalcin (N-MID), parathyroid hormone (PTH), vitamin D (VitD3), alkaline phosphatase (ALP), calcium (CA), phosphorus (P), cytokeratin 19 fragment (F211), and other indicators were performed. Four multiple regression models were established to determine the best diagnostic model for lung cancer with bone metastasis. RESULTS: Analysis of single indicators of bone metabolism markers in lung cancer was performed, among which F211, ß-CTX, TPINP, and ALP were significantly different (P < 0.05). The ROC curve of each indicator was less than 0.712. Based on the multiple regression models, the fourth model was the best and was much better than a single indicator with an AUC of 0.856, a sensitivity of 70.0%, a specificity of 91.0%, a positive predictive value of 82.5%, and a negative predictive value of 72.0%. CONCLUSION: Multiple regression models of bone metabolism markers were established. These models can be used to evaluate the progression of lung cancer and provide a basis for the early treatment of bone metastases.

[Feasibility of Preheating at 41 ℃ to Correct Red Blood Cell Parameters in the Presence of High-titer Cold Agglutinins].

Objective To explore the feasibility of preheating in 41 ℃ water bath for 30 minutes to correct the red blood cell parameters in the specimens containing high-titer cold agglutinins(CAs). Methods Two specimens containing high-titer CAs were selected during work,and the parameters of complete blood count at room temperature or after preheating in 37 ℃ or 41 ℃ water bath were compared.The smears were stained,and the distribution of red blood cells was observed with a microscope.Further,74 specimens without CAs were collected for complete blood count,and then the test results at room temperature and after preheating at 41 ℃ were compared. Results At room temperature,the specimens containing high-titer CAs showed significantly reduced red blood cell count(RBC)and hematocrit(HCT),abnormally increased mean corpuscular hemoglobin(MCH)and mean cell hemoglobin concentration(MCHC),abnormal percents of hemoglobin(HGB)and RBC,and aggregation of a large number of red blood cells.After being preheated at 37 ℃ for a certain time,the specimens demonstrated obviously improved parameters while still aggregation of a small number of red blood cells.After being preheated at 41 ℃ for 30 minutes,the specimens showed significantly increased RBC,normal HCT,MCH,and MCHC,and evenly distributed red blood cells.The 74 specimens without CAs showed the comparability was ≥80% between room temperature and preheating at 41 ℃ for 30 minutes or 60 minutes. Conclusion We can preheat the specimens containing high-titer CAs in a water bath at 41 ℃ to obtain accurate red blood cell parameters.

Functionally impaired follicular helper T cells induce regulatory B cells and CD14+ human leukocyte antigen-DR- cell differentiation in non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) represents one of the most common and aggressive cancers worldwide, as it typically displays irreversible progression and poor prognosis. Interaction between programmed death 1 (PD-1) and its ligand, PD-L1, plays important roles in tumor immunology. Follicular helper T (Tfh) cells have characteristically high PD-1 expression; thus, in the present study, we investigated the role of circulating Tfh cells and their correlation with disease-free survival after tumor resection in NSCLC. We found significantly higher number of Tfh cells but lower serum interleukin (IL)-21 levels in NSCLC patients, especially in those with advanced stage (III and IV), indicating that the function of Tfh cells to produce IL-21 was impaired. Further analysis showed that the increase in Tfh cells was attributable to an expansion of the PD-1+ -Tfh2 and PD-1+ -Tfh17 subtypes. Functional analysis showed that Tfh cells from NSCLC patients induced the differentiation of regulatory B cells and CD14+ human leukocyte antigen (HLA)-DR- cells. Interestingly, the number of Tfh1 subtypes in NSCLC patients was negatively correlated with disease-free survival after tumor resection. In short, the high number and abnormal function of Tfh cells could cause further immunosuppression and lead to tumor development in NSCLC. Rescuing Tfh functions therefore represents a potential therapeutic strategy in NSCLC.

Quality specifications of routine clinical chemistry methods based on sigma metrics in performance evaluation.

INTRODUCTION: Sigma metrics were applied to evaluate the performance of 20 routine chemistry assays, and individual quality control criteria were established based on the sigma values of different assays. METHODS: Precisions were expressed as the average coefficient variations (CVs) of long-term two-level chemistry controls. The biases of the 20 assays were obtained from the results of trueness programs organized by National Center for Clinical Laboratories (NCCL, China) in 2016. Four different allowable total error (TEa) targets were chosen from biological variation (minimum, desirable, optimal), Clinical Laboratory Improvements Amendments (CLIA, US), Analytical Quality Specification for Routine Analytes in Clinical Chemistry (WS/T 403-2012, China) and the National Cholesterol Education Program (NECP). RESULTS: The sigma values from different TEa targets varied. The TEa targets for ALT, AMY, Ca, CHOL, CK, Crea, GGT, K, LDH, Mg, Na, TG, TP, UA and Urea were chosen from WS/T 403-2012; the targets for ALP, AST and GLU were chosen from CLIA; the target for K was chosen from desirable biological variation; and the targets for HDL and LDL were chosen from the NECP. Individual quality criteria were established based on different sigma values. CONCLUSIONS: Sigma metrics are an optimal tool to evaluate the performance of different assays. An assay with a high value could use a simple internal quality control rule, while an assay with a low value should be monitored strictly.

Establishment of reference intervals for serum thyroid-stimulating hormone, free and total thyroxine, and free and total triiodothyronine for the Beckman Coulter DxI-800 analyzers by indirect method using data obtained from Chinese population in Zhejiang Province, China.

BACKGROUND: In order to establish suitable reference intervals of thyroid-stimulating hormone (TSH), free (unbound) T4 (FT4), free triiodothyronine (FT3), total thyroxine (T4), and total triiodothyronine (T3) for the patients collected in Zhejiang, China, an indirect method was developed using the data from the people presented for routine health check-up. METHODS: Fifteen thousand nine hundred and fifty-six person's results were reviewed. Box-Cox or Case Rank was used to transform the data to normal distribution. Tukey and Box-Plot methods were used to exclude the outliers. Nonparametric method was used to establish the reference intervals following the EP28-A3c guideline. Pearson correlation was used to evaluate the correlation between hormone levels and age, while Mann-Whitney U test was employed for quantification of concentration differences on the people who are younger and older than 50 years old. RESULTS: Reference intervals were 0.66-4.95 mIU/L (TSH), 8.97-14.71 pmol/L (FT4), 3.75-5.81 pmol/L (FT3), 73.45-138.93 nmol/L (total T4), and 1.24-2.18 nmol/L (total T3) in male; conversely, reference intervals for female were 0.72-5.84 mIU/L (TSH), 8.62-14.35 pmol/L (FT4), 3.59-5.56 pmol/L (FT3), 73.45-138.93 nmol/L (total T4), and 1.20-2.10 nmol/L (total T3). FT4, FT3, and total T3 levels in male and FT4 level in female had an inverse correlation with age. Total T4 and TSH levels in female were directly correlated. Significant differences in these hormones were also found between younger and older than 50 years old except FT3 in female. CONCLUSIONS: Indirect method can be applied for establishment of reference intervals for TSH, FT4, FT3, total T4, and total T3. The reference intervals are narrower than those previously established. Age factor should also be considered.

[Drug resistance and protoporphyrin ferrochelatase of Ralstonia mannitolilytica].

OBJECTIVE: To investigate the drug resistance, ß-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding genes of Ralstonia mannitolilytica, and to explore its structure and pathogenic function. METHODS: The strain was isolated by plate streaking method and identified by automatic bacteria detection system and 16S RNA gene PCR. Microdilution method was applied for drug susceptibility test. ß-lactamases, extended spectrum ß-lactamases (ESBL) and carbapenemases were detected using nitrocefin-disk, Kirby-Bauer disk, and Hodge test, respectively. Five ß-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding gene of the isolate were amplified by PCR for sequencing. Bioinformatic softwares were used to analyze the structure and function of the product of protoporphyrin ferrochelatase-encoding gene. RESULTS: A strain belonging to Ralstonia mannitolilytica was isolated. This isolate was sensitive to cefepime, ciprofloxacin, ofloxacin and tigecycline, but resistant to five penicillins, four cephalosporins and two carbapenems antibiotics. The isolate produced ß-lactamases but did not produce ESBL and carbapenemases. The isolate had five distinct ß-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding gene. The product of protoporphyrin ferrochelatase-encoding gene contained two functional domains of protoporphyrin ferrochelatase belonging to type Ⅱ chelatase superfamily that presented the most closely genetic relationship with the protoporphyrin ferrochelatase of Neisseria meningidis. CONCLUSIONS: The isolate of Ralstonia mannitolilytica has a higher resistance to ß-lactam antibiotics and its ß-lactamase-encoding genes are different with the common bacterial ß-lactamase-encoding genes. Protoporphyrin ferrochelatase may act as an important virulence factor of Ralstonia mannitolilytica.

A novel functional class 2 integron in clinical Proteus mirabilis isolates.

OBJECTIVES: To describe a novel functional class 2 integron that was found in clinical Proteus mirabilis isolates. METHODS: Class 1 and 2 integrons were screened by PCR in 153 clinical Proteus isolates. The variable regions of class 1 and 2 integrons were determined by restriction analysis and sequencing. The mutations of internal stop codons in class 2 integrons and their common promoters were also determined by sequencing. Enterobacterial repetitive intergenic consensus (ERIC)-PCR was used to analyse the phylogenetic relations of class 2 integron-positive P. mirabilis isolates. RESULTS: Class 1 integrons were detected in 96 (63%) of 153 Proteus isolates: eight different gene cassette arrays were detected, including dfrA32-ereA1-aadA2, which was detected for the first time in P. mirabilis. Class 2 integrons were detected in 101 (66%) of 153 Proteus isolates: four different gene cassette arrays were detected, including dfrA1-catB2-sat2-aadA1, which was detected for the first time in a class 2 integron. A novel functional class 2 integron was detected in 38 P. mirabilis isolates with a common promoter (-35 TTTAAT|16 bp|-10 TAAAGT). The variable region of this functional class 2 integron contained dfrA14 and three novel open reading frames with unknown functions. Very similar ERIC-PCR fingerprinting patterns were detected in these 38 P. mirabilis isolates and were different from other class 2 integron-positive isolates. CONCLUSIONS: A novel functional class 2 integron was found for the first time in P. mirabilis. These functional class 2 integron-harbouring P. mirabilis isolates were likely to be clonally spread in our hospital.

Synthesis and antitumor activity of feruloyl and caffeoyl derivatives.

We developed two efficient protocols for the synthesis of feruloyl and caffeoyl derivatives from commercial vanillin and veratraldehyde. Pharmacological activities were assessed against a panel of human cancer cell lines in vitro. Most synthesized compounds demonstrated attractive cytotoxicity. Several new compounds demonstrated significant antiproliferative and cytotoxic activities against HeLa and Bewo tumor cell lines. In particular, 5-nitro caffeic adamantyl ester showed broad spectrum of tumor inhibition in 10 cell lines, and reduced tumor weight by 36.7% in vivo when administered at a dose of 40 mg kg(-1).

[Screening and detection of patients with macroprolactinemia by application of polyethylene glycol precipitation method].

OBJECTIVE: To establish a polyethylene glycol (PEG6000) precipitation method for screening macroprolactinemia in patients with high serum prolactin (PRL). METHODS: PEG6000 precipitation method was used to remove macroprolactin (MPRL) molecules in serum of PRL-elevated patients. The effect of PEG6000 precipitating serum MPRL was determined by Sephacryl S-100HR chromatography plus chemiluminescent immunoassay and SDS-PAGE plus Western Blot assay. The PEG6000 precipitation plus chemiluminescent immunoassay was applied to screen serum samples of PRL-elevated patients for macroprolactinemia. The clinical manifestations of patients with true-hyperprolactinemia, hyperprolactinemia/macroprolactinemia or true-macroprolactinemia were analyzed and compared. RESULTS: After precipitation with PEG6000, MPRL peak or hybridization signal in the serum samples was markedly decreased, while the big or small prolactin (BPRL or SPRL) levels were not affected. In 1538 PRL-elevated patients, 16.1% (247/1538) were detectable for macroprolactinemia, while the 83.9% (1291/1538) were identified as true-hyperprolactinemia. In 247 samples of macroprolactinemia, 93.5% (231/247) were determined as true-macroprolactinemia, while 6.5% (16/247) were identified as hyperprolactinemia plus macroprolactinemia. In 508 true-hyperprolactinemia patients, menoxenia, menolipsis/menostasia, dysgenesia or hypophysoma were manifested in 438 (86.2%), which were also manifested in 85.7% (6/7) of hyperprolactinemia/macroprolactinemia patients. However, only 11 cases in 71 true-macroprolactinemia patients (15.5%) presented above clinical diseases. CONCLUSION: There is a certain proportion of true-macroprolactinemia (pseudo-hyperprolactinemia) in serum PRL-elevated patients. The PEG6000 precipitation method established in this study can efficiently distinguish true-hyperprolactinemia from pseudo-hyperprolactinemia in patients.

Expression of HLA-DR and KLRG1 enhances the cytotoxic potential and cytokine secretion capacity of CD3+ T cells in tuberculosis patients.

BACKGROUND: Human T cells play an important role in immunity against tuberculosis (TB) infection. Activating receptor HLA-DR and inhibitory receptor KLRG1 are critical regulators of T cell function during viral infection and tumorigenesis, but they have been less studied in TB infection. METHODS: In this study, we explored the relationship between CD3+ T cell expression of HLA-DR and KLRG1 receptors and function against TB infection. Flow cytometry was conducted to assess the immunomodulatory effects of HLA-DR and KLRG1 receptors on CD3+ T cells in patients with different TB infection status. RESULTS: We found activating receptors HLA-DR, NKG2C, CD57 and NKP46, and inhibitory receptors KLRG1 and KIR on CD3+ T cells in different TB infection status showed different distribution patterns; the cytotoxic potential and cytokine secretion capacity of CD3+ T cells after Mtb-specific antigen stimulation were significantly enhanced in TB infection groups. Further studies revealed HLA-DR+ T and KLRG1+ T cells expressed higher activating and inhibitory receptors than the negative population. In addition, the expression of cytotoxic potential and cytokine secretion capacity of HLA-DR+ T and KLRG1+ T cells was significantly higher than that of HLA-DR- T and KLRG1- T cells. CONCLUSIONS: Expression of HLA-DR and KLRG1 enhances the cytotoxic potential and cytokine secretion capacity of CD3+ T cells in TB patients, suggesting CD3+ T cells expressing HLA-DR and KLRG1 are important effector cell phenotypes involved in the host anti-TB infection. HLA-DR and KLRG1 expressed by CD3+ T cells may be potential predictive markers of TB disease progression and clinical immune assessment.

Differential diagnostic value of simultaneous detection of CD69 and HLA-DR on host T and NK cells in QFT-TB assay for identifying active tuberculosis.

BACKGROUND: Interferon-gamma release assay (IGRA) for tuberculosis (TB) remains limited in its ability to discriminate between active TB (ATB) and latent TB infection (LTBI). Activation markers on host T and NK cells are currently considered to be promising markers in the diagnosis of ATB. METHODS: This prospective observational study enrolled 213 participants and the participants were divided into ATB, LTBI, other lung-related diseases (ORD), and health control (HC) groups. CD69 and HLA-DR on T and NK cells were detected in QFT-TB assay, and a composite scoring system (TB-Flow) was created for the diagnosis of ATB. RESULTS: The expression of activation markers (CD69 and HLA-DR) were significantly increased in ATB. HLA-DR on NK cells, CD69 on T cells, and QFT-TB in the differential diagnosis of ATB and HC were all of good diagnostic value (AUC>0.90). In addition, the TB-Flow greatly improved the efficiency of differential diagnosis between ATB and LTBI (AUC=0.90, 95%CI: 0.84-0.96), with sensitivity and specificity of 79.17 % (95%CI: 64.60%-89.04 %) and 88.68 % (95%CI: 76.28%-95.31 %). CONCLUSIONS: CD69 and HLA-DR on host T and NK cells are promising markers in distinguishing different TB infection status. Our blood-based TB-Flow scoring system can distinguish ATB from LTBI with good diagnostic efficacy.

Diversity of gene cassette promoter variants of class 1 integrons in uropathogenic Escherichia coli.

Class 1 integrons play important roles in the emergence and horizontal transfer of antibiotic resistance genes among bacteria. The gene cassette promoter variants Pc or Pc-P2 of class 1 integrons not only drive the transcription of downstream gene cassettes, they also correlate with the excision and integration efficiency of the capture exogenous gene cassettes. In this study, the diversity of Pc or Pc-P2 variants of class 1 integrons and their association with antibiotic resistance phenotypes were analyzed in 132 uropathogenic Escherichia coli strains. Class 1 integrons were detected in 95 (72 %) strains. Sixteen different gene cassettes, 11 different gene cassette arrays and six different Pc or Pc-P2 variants were detected. The most prevalent gene cassettes were those that conferred resistance to trimethoprim, aminoglycosides, and chloramphenicol. The most prevalent promoter was PcH1, a relatively weak promoter. Certain gene cassette arrays or gene cassettes were mainly associated with the same Pc or Pc-P2 in different strains. Strains harboring class 1 integrons with relatively strong promoters had higher resistance rates to, or higher MIC(50) for, amikacin, chloramphenicol and tobramycin than those with relatively weak promoters. To the best of our knowledge, this is the first report of the diversity of class 1 integron Pc or Pc-P2 variants in uropathogenic E. coli strains.

Diagnostic Value of CD25, CD69, and CD134 on Tuberculosis-Specific Antigen-Stimulated CD4+ T Cells for Tuberculous Pleurisy.

Rapid and accurate methods for the diagnosis of tuberculous pleurisy (TP) are urgently needed. Activation markers of tuberculosis (TB)-reactive T cells are considered promising for the diagnosis of active TB (ATB). Different activation indexes may play different roles in the progression of TB, but there are few reports on T cell activation indicators, except for HLA-DR. Hence, we evaluated the expression of early (CD25 and CD69) and late (CD134) activation markers on TB antigen-stimulated CD4+ T cells in populations with different TB infection status and investigated their diagnostic value for ATB, particularly, for TP. Moreover, we compared the differences in the diagnostic efficacy among the indexes from peripheral blood (PB) and pleural fluid (PF) for TP. The expression of each activation marker was significantly increased in TB-infected populations (patients with ATB and latent TB infection vs. healthy individuals; patients with TP vs. non-TP) and was significantly higher in the PF than in the PB of patients with TP. The diagnostic performance of the coexpressed activation markers was superior to that of single expression markers in the differential diagnosis of ATB and non-TB, with CD25+CD134+ showing the best diagnostic efficiency (AUC: 0.93, 95% CI, 0.87-0.99; sensitivity: 86.7%, 95% CI, 72.5%-94.5%; and specificity: 94.0%, 95% CI, 82.5%-98.4%). Except for TB-IGRA, the activation indexes were more accurate than conventional laboratory methods for ATB diagnosis. In addition, the expression of CD25+CD134+ in PB and PF was the best values for differential diagnosis of TP and NTP, with AUCs of 0.87 (95% CI, 0.77-0.96) and 0.95 (95% CI, 0.90-1.00), respectively. Our study provides information on the diagnostic value of different activation markers for TB and shows that the expression of CD25+CD134+ on CD4+ T cells in PF can serve as a potential marker for TP diagnosis.

Host biomarkers other than interferon gamma in QFT-TB supernatants for identifying active tuberculosis.

Interferon gamma release assays (IGRAs) for tuberculosis (TB) remain limited in their ability to discriminate between active TB (ATB) and latent TB infection (LTBI). The objective of our study was to evaluate the value of additional cytokines/chemokines other than interferon gamma (IFN-γ) as biomarkers to identify different TB infection status. A total of 128 subjects were enrolled to detect the quantification of IL-2, IP-10, MCP-1 and RANTES in the supernatants of QuantiFERON®-TB (QFT-TB). Area under the curve (AUC) was used to evaluate the diagnostic efficiency. Notably, Mycobacterium tuberculosis (Mtb) induced cytokines/chemokines of ATB patients were significantly higher than those of the LTBI, other lung related diseases (ORD) and healthy controls (HC). Moreover, ROC analysis indicated that all cytokine/chemokine parameters detected were more capable of distinguishing ATB from LTBI than IFN-γ, especially IL-2. The diagnostic model including TB specific IL-2 and RANTES improved the performance in distinguishing ATB from LTBI, which was superior to single cytokines/chemokines in QFT-TB supernatants. Our results suggest that the combination of Mtb specific cytokines/chemokines has great prospects in the diagnosis of ATB, and the diagnostic model based on IL-2 and RANTES can be used as an alternative to distinguish ATB from LTBI.

Evaluating the diversity of circulating natural killer cells between active tuberculosis and latent tuberculosis infection.

Tuberculosis (TB) is a leading global public health problem; however, the mechanisms underlying the immunopathology of TB progression are not well understood. It is currently believed that Mycobacterium tuberculosis (Mtb) infection can modify natural killer (NK) cell phenotypic signatures. Hence, our study was designed to investigate the diversity of circulating NK cells in patients with different TB infection status. NK subsets, as well as their expression of activating and inhibitory receptors between active TB (ATB) and latent TB infection (LTBI) were evaluated. There were significant differences in NK cell phenotypes between ATB, LTBI and healthy controls. Notably, the proportion of KLRG1 in NK cells (P = 0.036), as well as in their subsets CD56DimCD16+ (P = 0.046) and CD27+ (P = 0.027) NK cells, increased significantly in LTBI group than in ATB group; while Mtb specific IFN-γ+CD56BrightCD16Dim NK cells expressed higher KLRG1 in ATB than in LTBI (P = 0.027). However, the expression of activating receptor NKG2D in NK subsets showed no significant difference among the study groups. Our results suggest that different TB infection status are coupled with the diversity of NK cell compartments, and the expression of KLRG1 in NK cells may be a specific phenotype that modulates the progression of TB from latent to active.

Serum Bile Acid Profiles in Latent Autoimmune Diabetes in Adults and Type 2 Diabetes Patients.

BACKGROUND: Impaired bile acid (BA) metabolism has been associated with the progression of type 2 diabetes (T2D). However, the contribution of BAs to the pathogenesis of latent autoimmune diabetes in adults (LADA) remains unclear. This study was aimed at investigating the association of serum BAs with different diabetes types and analyzing its correlation with main clinical and laboratory parameters. METHODS: Patients with LADA, patients with T2D, and healthy controls (HCs) were enrolled. Serum BA profiles and inflammatory cytokines were measured. The correlation of BA species with different indicators was assessed by Spearman's correlation method. RESULTS: Patients with diabetes (LADA and T2D) had significantly higher serum BAs, especially conjugated BAs, compared with those in HCs. Nevertheless, serum BA profiles had no special role in the progression of LADA, because no significant differences in BAs were observed between LADA and T2D patients. Interestingly, HbA1c levels and HOMA-ß were found to be correlated with a series of BA species. Proinflammatory cytokines (IL-1ß, IL-6, and TNF-α) and anti-inflammatory cytokine (IL-10) were all positively associated with several BA species, especially the conjugated secondary BAs. CONCLUSION: Serum BAs regulate glucose homeostasis, but have no special value in the pathogenesis of LADA patients. Our study adds further information about the potential value of serum BAs in different types of diabetes.