Multiple cytokine analysis based on QuantiFERON-TB gold plus in different tuberculosis infection status: an exploratory study.

BACKGROUND: More efficient and convenient diagnostic method is a desperate need to reduce the burden of tuberculosis (TB). This study explores the multiple cytokines secretion based on QuantiFERON-TB Gold Plus (QFT-Plus), and screens for optimal cytokines with diagnostic potential to differentiate TB infection status. METHODS: Twenty active tuberculosis (ATB) patients, fifteen patients with latent TB infection (LTBI), ten patients with previous TB and ten healthy controls (HC) were enrolled. Whole blood samples were collected and stimulated by QFT-Plus TB1 and TB2 antigens. The levels of IFN-γ, TNF-α, IL-2, IL-6, IL-5, IL-10, IP-10, IL-1Ra, CXCL-1 and MCP-1 in supernatant were measured by Luminex bead-based multiplex assays. The receiver operating characteristic curve was used to evaluate the diagnostic accuracy of cytokine for distinguishing different TB infection status. RESULTS: After stimulation with QFT-Plus TB1 and TB2 antigens, the levels of all cytokines, except IL-5 in TB2 tube, in ATB group were significantly higher than that in HC group. The levels of IL-1Ra concurrently showed the equally highest AUC for distinguishing TB infection from HC, followed by the levels of IP-10 in both TB1 tube and TB2 tube. Moreover, IP-10 levels displayed the largest AUC for distinguishing ATB patients from non-ATB patients. Meanwhile, the levels of IP-10 also demonstrated the largest AUC in both TB1 tube and TB2 tube for distinguishing ATB patients from LTBI. CONCLUSIONS: In addition to conventional detection of IFN-γ, measuring IP-10 and IL-1Ra based on QFT-Plus may have the more tremendous potential to discriminate different TB infection status.

Utility of interferon gamma/tumor necrosis factor alpha FluoroSpot assay in differentiation between active tuberculosis and latent tuberculosis infection: a pilot study.

BACKGROUND: The differential diagnosis of active tuberculosis (ATB) and latent tuberculosis infection (LTBI) remains challenging in clinical practice. We aimed to evaluate the diagnostic accuracy of the IFN-γ/TNF-α FluoroSpot assay for differentiating ATB from LTBI. METHODS: We conducted a pilot study of case-control design, using the FluoroSpot assay to simultaneously detect IFN-γ and TNF-α secretion at the single-cell level. The frequencies of antigen-specific single TNF-α-, total TNF-α-, single IFN-γ-, total IFN-γ- and dual IFN-γ/TNF-α-secreting T cells were detected. The optimal cutoffs value of frequencies for differentiating ATB from LTBI were determined according to receiver operating characteristic curve analysis. The sensitivity, specificity, predictive values (PV) and likelihood ratios (LR) of the FluoroSpot assay were calculated. RESULTS: Thirty patients diagnosed microbiologically with ATB, 36 healthcare workers with LTBI and 36 healthy controls were enrolled. After stimulated by ESAT-6 or CFP-10 peptides, the median frequencies of single TNF-α-, total TNF-α-, single IFN-γ-, total IFN-γ- and dual IFN-γ/TNF-α-secreting T cells in ATB patients were all significantly higher than those in LTBI and HC groups (P < 0.01). The frequencies of total IFN-γ-secreting T cells detected by FluoroSpot assay correlated significantly with those of T-SPOT.TB (r = 0.910 for ESAT-6, P < 0.001, r = 0.845 for CFP-10, P < 0.001). After stimulated by ESAT-6 peptides, with total TNF-α-secreting T cells frequencies at a cut off value of 21 iSFCs/250,000 PBMCs, the sensitivity, specificity, PLR, NLR, PPV, NPV of IFN-γ/TNF-α FluoroSpot assay in differentiating ATB from LTBI were 96.7% (95%CI, 82.8-99.9%), 94.3% (95%CI, 80.8-99.3%), 16.92 (95%CI, 4.40-65.08), 0.04 (95%CI, 0.01-0.24), 93.6% (95%CI,78.6-99.2%) and 97.1% (95%CI, 84.7-99.9%), respectively. With the frequencies of total TNF-α- and total IFN-γ-secreting T cells stimulated by ESAT-6 peptides combined, the specificity was increased to 97.1%, and the positive likelihood ratio to 31.5. The combination with CFP-10 might not improve the diagnostic accuracy of the ESAT-6 for differentiating ATB from LTBI. CONCLUSIONS: IFN-γ/TNF-α FluoroSpot assay might have potential to help differentiate ATB from LTBI, but the findings need to be further verified by cross-sectional or prospective cohort studies.

Risk assessment of soil heavy metals associated with land use variations in the riparian zones of a typical urban river gradient.

Urbanization-induced land use changes in riparian area alter soil and water regimes in complex ways, which may also affect the migration and transformation of soil heavy metals and increase the risk of release. In this study, soil samples from the riparian zone of Beiyun River, which located in the rapidly urbanized Beijing metropolis, were collected and analyzed for heavy metals (As, Cd, Cr, Cu, Mn, Ni, Pb, and Zn). Then their zoning distribution pattern along river (section 1 to section 4 from upper to low reaches) and the correlation of heavy metals between riparian soils and riverine sediments were investigated. Results showed that the average soil heavy metal concentrations of Cd, Cr, Cu and Zn in riparian zone were approximately 2.2, 1.7, 1.9 and 2.0 times higher than the background values. Sectionally, the concentrations of Cd, Ni, Pb and Zn displayed a decreasing order with section 2 > section 3 > section 4 > section 1, while the highest values of Cr and Cu were found in section 3. The concentrations of all heavy metals except Cr in artificial garden land were higher than those in other land use types, and the concentrations of Cr among five land use types were in the order of grass land > farmland > artificial garden land > forest land > forest-grass land. Generally, most of the heavy metals in the riverine sediments had higher contents than those in riparian zones, especially Cu and Zn. There was a decreasing order for the average geo-accumulation index (Igeo) of measured heavy metals in the soils of riparian zone: Zn (0.15) > Cr (0.08) > Cu (0.07) > Cd (-0.08) > As (-0.57) > Pb (-0.67) > Mn (-0.75) > Ni (-0.86), whereas they had different "high-low" orders in different land use types. The Igeo index indicated most regions of riparian zone were moderately polluted with Cd, Cr, Cu and Zn, especially in grass land and forest land. Also, Cd, Cr and Zn in riparian zone have positive relationships with the concentrations in riverine sediments. Health risk assessment showed that the contribution of ingestion HQ to HI was the highest among the three exposure pathways (ingestion, inhalation and dermal contact), and children had higher non-carcinogenic risk and carcinogenic risk index than adult. Our findings suggest that land use and soil in riparian zone should be protected and managed scientifically to control the riverine pollution and ensure human health.

Diagnostic Utility of Interferon-Gamma Release Assay in Tuberculous Lymphadenitis.

Objective The aim of this study was to evaluate the diagnostic performance of T-SPOT.TB for tuberculous lymphadenitis. Methods Suspected tuberculous lymphadenitis patients between September 2010 and September 2018 who had both peripheral blood T-SPOT.TB test and lymph node biopsy were retrospectively enrolled in this study. The cutoff value of T-SPOT.TB test for peripheral blood was set as 24 spot forming cell (SFC)/10 6 periphreral blood monocyte cell (PBMC) according to the instruction of testing kits. The gold standard for diagnosis of TBL was the combination of microbiology results, histopathology results and patient's response to anti-TB treatment. Diagnostic efficacy of T-SPOT.TB was evaluated, including sensitivity, specificity, accuracy, predictive values, and likelihood ratio. Results Among 91 patients who met the inclusion criteria, we excluded 8 cases with incomplete clinical information and 6 cases who lost to follow-up. According to the gold standard, there were 37 cases of true TBL (9 confirmed TBL and 28 probable TBL), 30 cases of non-TBL, and 10 cases of clinically indeterminate diagnosis who were excluded from the final analyses. The T-SPOT.TB tests yielded 43 cases of positive response and 24 cases of negative response. The sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR) and negative likelihood ratio (NLR) of peripheral blood T-SPOT.TB for diagnosing TBL were 89.2%, 66.7%, 79.1%, 76.7%, 83.3%, 2.68 and 0.16, respectively. The number of SFCs of T-SPOT.TB in TBL patients [432(134-1264)/10 6 PBMCs] was higher than that in non-TBL patients [0 (0-30) /10 6PBMCs] with a significant difference (Z=-5.306, P <0.001).Conclusion T-SPOT.TB is a rapid and simple diagnostic test for TBL with a high sensitivity and negative predictive value.

Spatial Distribution of Metals and Associated Risks in Surface Sediments Along a Typical Urban River Gradient in the Beijing Region.

Surface sediments from Beiyun River located in the rapidly urbanized Beijing metropolis were collected and analyzed for heavy metals (As, Cd, Cr, Cu, Mn, Ni, Pb, and Zn) to investigate their spatial distribution pattern, ecotoxicology and source identification. Results indicated the average heavy metal concentrations of Cd, Cr, Cu, Pb, and Zn were approximately 4, 2, 3, 2, and 4 times higher than their background values. Spatially, we found the concentrations of heavy metals made significant change in four sections along urbanized river gradients. The contents in midstream of urban region and farmland region (section 2 and section 3) were greater than those in upstream (section 1) and downstream (section 4). However, one-way analysis of variance for spatial analysis suggested there were no significant differences between mainstream and tributaries. The geo-accumulation index (I geo) used to assess the sediment quality exhibited there was a decreasing order for the average I geo of measured heavy metals: Zn (0.82) > Cd (0.53) > Cu (0.50) > Cr (- 0.08) > Pb (- 0.45) > Ni (- 0.96) > Mn (- 0.97) > As (- 1.01), whereas they had different "high-low" orders at different sampling transects. Ecological risk index values showed that section 2 and section 3 revealed a high and moderate ecological risk, respectively. Furthermore, principal component analysis indicated the first principle component explained 64.73% of total variance with the main pollutants of As, Cr, Ni, Zn, and Cu which were probably controlled by the mixed sources covering natural factors and anthropogenic input.

The characteristics of patients with mycobacterium tuberculosis blood stream infections in Beijing, China: a retrospective study.

BACKGROUND: Published information regarding the clinical characteristics, laboratory findings, and outcomes of patients with Mycobacterium tuberculosis (MTB) blood stream infection (BSI) is limited. We aimed in this study to evaluate the clinical characteristics, laboratory evaluation, and outcomes of patients with MTB BSI. METHODS: All patients diagnosed with MTB BSI at Peking Union Medical College Hospital between January 2008 and May 2014 were identified by examining the electronic database listing results of all blood cultures. Data on demographics, clinical characteristics, laboratory manifestations, management, and outcomes were abstracted from medical records. RESULTS: Six thousand nine hundred seventy-four patients had mycobacterial blood cultures during the study period. Of 48 patients (0.7%) with MTB BSI, 26 patients (54%) were considered to be immunocompromised (refers to a person who has a significantly impaired immune system). This was due to human immunodeficiency virus (HIV) infection (n = 2 of 48 tested), receiving steroids (n = 17, including 16 with rheumatic diseases and one with myasthenia gravis), malignancy (n = 3), diabetes mellitus (n = 3), and renal transplantation (n = 1). The main clinical manifestations were fever (100%, with a median of 40 °C), weight loss (48%) and cough with sputum production (46%). Most patients had one or more organs involved (81%). The median time from onset of fever to diagnosis was 8 weeks (IQR 5 ~ 14). Six patients died within 1 week after diagnosis. Of the 17 patients completing treatment, 14 patients (82%) recovered without major complications and they had a shorter time interval between onsets of symptoms to treatment compared to those died of TB. CONCLUSIONS: In this group of patients with MTB BSI, fever and multiple organs involvement were common, the outcome was poor and timely diagnosis and treatment might favor outcome.

Positive Rate of Different Hepatitis B Virus Serological Markers in Peking Union Medical College Hospital, a General Tertiary Hospital in Beijing.

Objective s To investigate the positive rate of different hepatitis B virus (HBV) serological markers, and the demographic factors related to HBV infection.Methods We enrolled all patients tested for HBV serological markers, such as HBV surface antigen (HBsAg), HBV surface antibody (HBsAb), hepatitis B e antigen (HBeAg), hepatitis B e antibody (HBeAb), HBV core antibody (HBcAb), and HBV-DNA from July 2008 to July 2009 in Peking Union Medical College Hospital. The positive rate of each HBV serological marker was calculated according to gender, age, and de- partment, respectively. The positive rates of HBV-DNA among patients with positive HBsAg were also analyzed.Results Among 27 409 samples included, 2681 (9.8%) were HBsAg positive. When patients were divided into 9 age groups, the age-specific positive rate of HBsAg was 1.2%, 9.6%, 12.3%, 10.9%, 10.3%, 9.7%, 8.0%, 5.8%, and 4.3%, respectively. The positive rate of HBsAg in non-surgical department, surgical department, and health examination center was 16.2%,5.8%,and 4.7%, respectively. The positive rate of HBsAg of males (13.3%) was higher than that of females (7.3%, P=0.000). Among the 2681 HBsAg (+) patients, 1230 (45.9%) had HBV-DNA test, of whom 564 (45.9%) were positive. Patients with HBsAg (+), HBeAg (+), and HBcAg (+) result usually had high positive rate of HBV-DNA Results (71.8%, P=0.000).Conclusions Among this group of patients in our hospital, the positive rate of HBsAg was relatively high. Age group of 20-29, males, and patients in non-surgical departments were factors associated with high positive rate of HBsAg.

A single-institution retrospective evaluation of noninvasive localization for non-palpable breast microcalcification.

TECHNIQUE: From January 1, 2018, to December 31, 2021, we localized the breast microcalcification of 40 patients before the surgical excision. We measured the distance between the nipple and the center of the calcification on the CC view and the ML view, respectively. The operation proceeded around the intersection between two lines, slightly larger than the diameter of the microcalcification. We also analyze the pathological findings. RESULTS: All 40 patients successfully detected calcification by mammograms preoperatively using the method mentioned above. 38 patients have the microcalcification removal within the one-time operation, while the other two underwent an extended lumpectomy. 20 of 40 calcifications (50 %) were malignant and 12(30 %) were precancerous lesions. In the group of women older than 45 years old, the percentages of malignant and atypical hyperplasias are 56.25 % (18/32) and 31.25 % (10/32) respectively. CONCLUSION: Our non-invasive method of preoperative localization is safe and cost-effective. Furthermore, initial observations suggest that there may be a link between age and malignant microcalcification.

Specific Cytokines Analysis Incorporating Latency-Associated Antigens Differentiates Mycobacterium tuberculosis Infection Status: An Exploratory Study.

Introduction: Current immunologic methods cannot distinguish Mycobacterium tuberculosis (Mtb) infection statuses, especially to discriminate active tuberculosis (ATB) from latent tuberculosis infection (LTBI). This study explored the potential of latency-associated antigens (Rv1733cSLP and Rv2028c) and multifactorial cytokine detection to distinguish tuberculosis infection states. Methods: ATB patients (20), LTBI healthcare workers (25), fever patients (11), and healthy controls (10) were enrolled. Cytokine levels (IFN-γ, TNF-α, IL-2, IL-6, IP-10, IL-1Ra, CXCL-1, and MCP-1) were measured using Luminex with/without MTB-specific virulence factor and latency-associated antigens stimulation. Results: Without antigen stimulation, IL-6, IP-10, MCP-1, and IL-1Ra were higher in the ATB group than in the LTBI group (p<0.05), but no significant differences between the ATB group and the fever group. Stimulated with the four antigens, respectively, the cytokines, including IP-10Esat-6, IP-10CFP-10, IFN-γRv1733cSLP, IFN-γRv2028c, IL-6Esat-6, IL-6Rv1733cSLP, IL-6Rv2028c, IL-2Rv1733cSLP, IL-2 Rv2028c, IL-1RaEsat-6, IL-1RaCFP-10, IL-1RaRv2028c, CXCL-1Esat-6, CXCL-1CFP-10, CXCL-1Rv1733cSLP, CXCL-1Rv2028c, MCP-1Esat-6 and MCP-1CFP-10, demonstrated accurate discrimination between ATB and LTBI (p<0.05). Additive concentrations demonstrated significant secretion differences of IFN-γ, IP-10 and IL-2, primarily by virulence factors in ATB and latency-associated antigens in LTBI. Latency-associated antigens synergized with virulence factors, enhancing TH1-type cytokine diagnostic efficacy for discriminating ATB from LTBI, the AUC for TNF-α increased from 0.696 to 0.820 (p=0.038), IFN-γ increased from 0.806 to 0.962 (p=0.025), and IL-2 increased from 0.565 to 0.868 (p=0.007). Model selected by forward likelihood method indicated combined detection of IFN-γCFP-10, IFN-γRv1733cSLP, IP-10Rv1733cSLP, and CXCL-1Rv1733cSLP achieved ATB diagnosis (AUC=0.996) and ATB-LTBI differentiation (AUC=0.992). Combined detection of IFN-γCFP-10 and IFN-γRv1733cSLP achieved tuberculosis infection diagnosis (AUC=0.943). Conclusion: Latency-associated antigens enhance multiple cytokine discriminatory ability, particularly TH1-type cytokines, for differentiating Mtb infection statuses.

Latent Tuberculosis Infection and Associated Factors in Patients with Systemic Lupus Erythematosus: a Multicenter, Cross-Sectional Study.

The objectives of this study were to screen for latent tuberculosis infection (LTBI) among patients with systemic lupus erythematosus (SLE) using the T-SPOT.TB assay and to identify factors affecting the assay results. SLE patients were enrolled from 13 tertiary hospitals in eastern, central, and western China from September 2014 to March 2016 and were screened using the T-SPOT.TB assay to detect LTBI. Basic information about the subjects was collected, including gender, age, body mass index (BMI), course of disease, evidence of previous tuberculosis, Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K) score, and the use of glucocorticoids and immunosuppressants. Univariate analysis and multivariable logistic regression were performed to identify factors affecting the results of the T-SPOT.TB assay. In all, 2,229 SLE patients were screened using the T-SPOT.TB assay, of whom 334 patients tested positive, yielding a positivity rate of 15% (95% confidence interval [CI], 13.5% to 16.5%). The positivity rate was higher in male than female patients and had an increasing trend with age. Multivariable logistic regression analysis showed that patients over 40 (odds ratio [OR], 1.65; 95% CI, 1.29 to 2.10) and with evidence of previous tuberculosis (OR, 4.43; 95% CI, 2.81 to 6.99) were more likely to have positive T-SPOT.TB results, while patients with a SLEDAI-2K score of ≥10 (OR, 0.61; 95% CI, 0.43 to 0.88), a glucocorticoid dose of ≥60 mg/d (OR, 0.62; 95% CI, 0.39 to 0.98), leflunomide (LEF) treatment (OR, 0.51; 95% CI, 0.29 to 0.88), or tacrolimus (FK506) treatment (OR, 0.40; 95% CI, 0.16 to 1.00) were more likely to have negative T-SPOT.TB results. The frequencies of CFP-10-specific gamma interferon (IFN-γ)-secreting T cells were significantly lower in SLE patients with severe disease activity or high-dose glucocorticoids (P < 0.05). The positivity rate of the T-SPOT.TB assay was 15% among SLE patients. Severe, active SLE disease and the use of high-dose glucocorticoids and some types of immunosuppressants are likely to result in negative T-SPOT.TB results. For SLE patients with the above conditions, diagnosing LTBI based on a positive T-SPOT.TB result may lead to underestimation of the prevalence. IMPORTANCE The burden of tuberculosis and systemic lupus erythematosus in China ranks among the top three in the world. Therefore, active screening for LTBI and preventive intervention in SLE patients are of great significance in China. In view of the lack of relevant data in a large sample, we conducted a multicenter, cross-sectional study using T-SPOT.TB as a screening method for LTBI, to investigate the prevalence of LTBI and analyze the factors affecting the results of the T-SPOT.TB assay in SLE patients. Our study showed that the overall positivity rate of the T-SPOT.TB assay in SLE patients was 15.0%, which was lower than the estimated LTBI prevalence in the general population in China (~20%). For SLE patients with severe, active disease, high-dose glucocorticoids, and some types of immunosuppressants, a diagnosis of LTBI based on only positive T-SPOT.TB results may lead to underestimation of the prevalence.

Mycobacterium tuberculosis latency-associated antigen Rv1733c SLP improves the accuracy of differential diagnosis of active tuberculosis and latent tuberculosis infection.

BACKGROUND: Differential diagnosis of active tuberculosis (ATB) and latent tuberculosis infection (LTBI) has been a challenge for clinicians in high TB burden countries. The purpose of this study was to improve the accuracy of differential diagnosis of ATB and LTBI by using fluorescent immunospot (FluoroSpot) assay to detect specific Th1 cell immune responses. The novel mycobacterium tuberculosis (MTB) latency-associated antigens Rv1733c and synthetic long peptides derived from Rv1733c (Rv1733c SLP) were used based on virulence factors early secreting antigen target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10). METHODS: Fifty-seven ATB cases, including 20 pathogen-confirmed ATB and 37 clinically diagnosed ATB, and 36 LTBI cases, were enrolled between January and December 2017. FluoroSpot assay was used to detect the interferon γ (IFN-γ) and interleukin 2 (IL-2) secreted by the specific T cells after being stimulated with MTB virulence factors ESAT-6 and CFP-10, MTB latency-associated antigens Rv1733c and Rv1733c SLP. The receiver operating characteristic (ROC) curve was used to define the best cutoff value of latency-associated antigens in the use of differentiating ATB and LTBI. The sensitivity, specificity, predictive value, and likelihood ratio of ESAT-6 and CFP-10-FluoroSpot combined with latency-associated antigen in the differential diagnosis of ATB and LTBI were also calculated. RESULTS: Following the stimulation with Rv1733c and Rv1733c SLP, the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP had the largest area under the ROC curve, which was 0.766. With a cutoff value of 1 (spot-forming cells [SFCs]/2.5 × 105 peripheral blood mononuclear cells) for frequency, the sensitivity and specificity of distinguishing ATB from LTBI were 72.2% and 73.7%, respectively. ESAT-6 and CFP-10-FluoroSpot detected the frequency and proportion of single IFN-γ-secreting T cells; the sensitivity and specificity of distinguishing ATB from LTBI were 82.5% and 66.7%, respectively. Combined with the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP on the basis of ESAT-6 and CFP-10-FluoroSpot, the sensitivity and specificity increased to 84.2% and 83.3%, respectively. CONCLUSION: Rv1733c SLP, combined with ESAT-6 and CFP-10, might be used as a candidate antigen for T cell-based tuberculosis diagnostic tests to differentiate ATB from LTBI.

Comparison of diagnostic accuracy of QuantiFERON-TB Gold Plus and T-SPOT.TB in the diagnosis of active tuberculosis in febrile patients.

OBJECTIVE: This study aimed to compare the accuracy of QuantiFERON-TB Gold Plus (QFT-Plus) and T-SPOT.TB for diagnosing active tuberculosis (ATB) in febrile patients, to explore influencing factors of positive results and to verify the potential value of QFT-Plus in the identification of ATB and latent tuberculosis infection (LTBI). METHODS: A total of 240 febrile patients with ATB (n = 80) and non-ATB (n = 160) were recruited to assess the accuracy of QFT-Plus and T-SPOT.TB for diagnosing ATB. Multivariable logistic regression was used to analyze the influencing factors of positive results. RESULTS: The proportion of indeterminate results (ITRS) in QFT-Plus and T-SPOT.TB were 3.3% and 0%, respectively. The consistency between the results of the QFT-Plus and T-SPOT.TB was substantial. The area under the receiver operating characteristic curve (AUROC) of the QFT-Plus and T-SPOT.TB for diagnosing ATB was 0.792 and 0.849 (p = 0.070), respectively. The sensitivity of differentiating ATB from non-ATB was 92.2% in QFT-Plus versus 95.0% in T-SPOT.TB. The influencing factors of T-SPOT.TB positive result were male (odds ratio (OR) = 2.33, 95% confidence interval (CI) 1.27-4.26, p = 0.006), evidence of previous TB (OR 11.36, 95% CI 4.62-27.94, p < 0.001), while male (OR = 3.17, 95% CI 1.73-5.84, p < 0.001), evidence of previous TB (OR = 7.58, 95% CI 3.60-15.98, p <0.001), and use of immunosuppressant (OR = 0.49, 95% CI 0.260.94, p = 0.030) were influencing factors for QFT-Plus positive result. There was no significant difference in QFT-Plus in differentiating ATB from LTBI in febrile patients. CONCLUSION: There was no significant difference between QFT-Plus and T-SPOT.TB for diagnosing ATB in febrile patients. QFT-Plus is prone to ITRS. The influencing factors including males, evidence of the previous TB, and use of immunosuppressant should be considered when interpreting positive results.

Prevalence and risk factors of active tuberculosis in patients with rheumatic diseases: a multi-center, cross-sectional study in China.

Evidence of active tuberculosis (ATB) in patients with rheumatic diseases are research priorities but limited data from China have been reported. Research targeting patients not taking anti-TNF biologics are especially insufficient. We aimed to investigate the prevalence and risk factors of ATB in this at-risk population. We conducted a tertiary hospital-based, multi-center, cross-sectional study by using stratified multi-stage cluster sampling strategy to screen ATB in patients with rheumatic diseases. We estimated the prevalence of ATB in patients with rheumatic diseases and identified risk factors among those who were not taking anti-TNF biologic. A total of 13,550 eligible patients were enrolled, and the result showed the standardized prevalence of ATB according to the composition ratio of various types of rheumatic disease was 882/100000 (95% confidence interval (CI): 706-1057). Multivariable logistic regression analysis in patients not taking anti-TNF biologics showed that the independent risk factors of ATB were having systemic lupus erythematosus (SLE) (OR=2.722, 95% CI: 1.437-5.159, p=0.002), having Behcet's disease (BD) (OR= 5.261, 95% CI: 2.071-13.365, p<0.001), taking azathioprine(AZA) within the past two years (OR=2.095, 95% CI: 0.986-4.450, p=0.054), exposing to glucocorticoids ≥30mg/d for more than four weeks within the past two years (OR=2.031, 95% CI: 1.247-3.309, p=0.004) and having evidences of previous TB (OR= 6.185, 95% CI: 3.487-10.969, p<0.001). The prevalence of ATB was higher in patients with rheumatic diseases compared to the general population. Patients with SLE or BD, prolonged exposure to moderate to high dose of glucocorticoids and previous TB were independent risk factors for ATB.

The establishment of CMV antigen-specific enzyme-linked immunospot assay among Chinese: A pilot study.

BACKGROUND: The prevalence of cytomegalovirus in China is high and the infection could result in disastrous consequences. Cellular immunity is the main mechanism for virus controlling. We explored the performance of cytomegalovirus antigen-specific enzyme-linked immunospot assay in patients with different infection states in endemic area. METHODS: Twenty-eight patients with reliable results were included in statistical analysis. Peripheral blood mononuclear cells were extracted and were stimulated by cytomegalovirus phosphoprotein 65 or immediate early protein-1 antigen respectively. Spot forming cells (SFCs) were counted to evaluate the cellular immune response elicited by antigens. RESULTS: Spots could be clearly displayed, and evenly dispersed with a clean background. The numbers of SFCs were 0 [0-0], 426 [210-601] and 230 [57-513] for uninfected individuals (n = 7), latently infected patients (n = 11) and actively infected patients (n = 10) respectively, which were statistically different. The number of SFCs stimulated by phosphoprotein 65 was significantly higher than that by immediate early protein-1. CONCLUSIONS: The number of SFCs was significantly different among patients with different infection state. The stimulatory effect of phosphoprotein 65 was better than that of immediate early protein-1.

Application of IFN-γ/IL-2 FluoroSpot assay for distinguishing active tuberculosis from non-active tuberculosis: A cohort study.

Currently available Interferon-γ release assay cannot reliably differentiate active TB (ATB) from non-active TB (non-ATB). This study aimed to evaluate the diagnostic accuracy of the IFN-γ/IL-2 FluoroSpot assay, which can simultaneously detect IFN-γ and IL-2 secretion, for differentiating ATB from non-ATB. 191 suspected ATB patients with positive T-SPOT.TB results were consecutively recruited. 64 (33.5%) participants had ATB, including 22 (34.4%) microbiologically or histologically confirmed TB and 42 (65.6%) clinically diagnosed TB. 119 (62.3%) cases were non-ATB and 8 (4.2%) were clinically indeterminate. After being stimulated with ESAT-6 and CFP-10 antigens, the median frequency and proportion of IFN-γ+IL-2- T cells were significantly higher in the ATB group than the non-ATB group (P < .001). The areas under the ROC curves of IFN-γ+IL-2- T cells were larger than those of total IFN-γ+ T cells (0.788 vs. 0.739, p = .323). With a cutoff value of 25 SFCs/250,000 PBMCs for frequency, sensitivity and specificity of this assay were 73.4% and 69.8% respectively. When combining the frequency and proportions of IFN-γ+IL-2- T cells, the sensitivity and specificity were increased to 95.3% in parallel testing and 83.2% in serial testing respectively. In conclusion, IFN-γ/IL-2 FluoroSpot assay is conducive for the diagnosis of ATB in patients with positive T-SPOT.TB results.

Prevalence and influencing factors of the high nil-control spot count in T-SPOT.TB: A matched case-control study.

BACKGROUND: T-SPOT.TB may yield indeterminate results, including high nil responses and insufficient mitogen responses. We explored the incidence and risk factors of high nil responses. METHODS: A 1:1 matched case-control study of patients who underwent T-SPOT.TB tests in Peking Union Medical College Hospital from Jan 1, 2015 to Apr 30, 2017 was conducted. High nil responses were defined as >10 spots in negative control wells. Cases and controls were matched based on when the tests were performed. Demographic, clinical and laboratory data were obtained from the Medical Record System. RESULTS: A total of 644 out of 36,316 (1.76%, 95% CI: 1.63%-1.90%) patients presented with high nil responses (280 cases and 280 controls were enrolled). Multivariate analysis revealed that male (OR = 1.882, 95% CI: 1.222-2.899), Behcet's disease (OR = 7.764, 95% CI: 1.714-35.167), heavy use of corticosteroids within a month (OR = 0.357, 95% CI: 0.138-0.921, for >1000 mg group) and hypoalbuminemia (OR = 0.385, 95% CI: 0.241-0.615) are significantly associated with high nil responses. CONCLUSIONS: High nil responses in T-SPOT.TB assays are quite rare. Male gender and Behcet's disease are suggested as independent risk factors, while recent excessive use of corticosteroids and hypoalbuminemia seem to be independent protective factors.

A regional strategy for ecological sustainability: A case study in Southwest China.

Partitioning, a method considering environmental protection and development potential, is an effective way to provide regional management strategies to maintain ecological sustainability. In this study, we provide a large-scale regional division approach and present a strategy for Southwest China, which also has extremely high development potential because of the "Western development" policy. Based on the superposition of 15 factors, including species diversity, pattern restriction, agricultural potential, accessibility, urbanization potential, and topographical limitations, the environmental value and development benefit in the region were quantified spatially by weighting the sum of indicators within environmental and development categories. By comparing the scores with their respective median values, the study area was divided into four different strategy zones: Conserve zones (34.94%), Construction zones (32.95%), Conflict zones (16.96%), and Low-tension zones (15.16%). The Conflict zones in which environmental value and development benefit were both higher than the respective medians were separated further into the following 5 levels: Extreme conflict (36.20%), Serious conflict (28.07%), Moderate conflict (12.28%), Minor conflict (6.55%), and Slight conflict (16.91%). We found that 9.04% of nature reserves were in Conflict zones, and thus should be given more attention. This study provides a simple and feasible method for regional partitioning, as well as comprehensive support that weighs both the environmental value and development benefit for China's current Ecological Red Line and space planning and for regional management in similar situations.

Identifying trace metal distribution and occurrence in sediments, inundated soils, and non-flooded soils of a reservoir catchment using Self-Organizing Maps, an artificial neural network method.

The Lancang-Mekong River is a trans-boundary river which provides a livelihood for over 60 million people in Southeast Asia. Its environmental security is vital to both local and regional inhabitants. Efforts have been undertaken to identify controlling factors of the distribution of trace metals in sediments and soils of the Manwan Reservoir catchment in the Lancang-Mekong River basin. The physicochemical attributes of 63 spatially distributed soil and sediment samples, along with land-use, flooding, topographic, and location characteristics, were analyzed using the Self-Organizing Map (SOM) methodology. The SOM permits the analysis of complex multivariate datasets and gives a visual interpretation that is generally not easy to obtain using traditional statistical methods. Across the catchment, enrichments of trace metals are rare overall, despite the severely enriched cadmium (Cd). The analysis of SOM showed that flooded levels and land-use types were associated with high concentrations of Cd. Sediments and inundated soils covered with shrub and open woodlands in downstream always have a high concentration of Cd. The results demonstrate that SOM is a useful tool that can aid in the interpretation of complex datasets and help identify the environment of enriched metals on a catchment scale.

Analysis of Factors Influencing Diagnostic Accuracy of T-SPOT.TB for Active Tuberculosis in Clinical Practice.

T-SPOT.TB didn't perform a perfect diagnosis for active tuberculosis (ATB), and some factors may influence the results. We did this study to evaluate possible factors associated with the sensitivity and specificity of T-SPOT.TB, and the diagnostic parameters under varied conditions. Patients with suspected ATB were enrolled prospectively. Influencing factors of the sensitivity and specificity of T-SPOT.TB were evaluated using logistic regression models. Sensitivity, specificity, predictive values (PV), and likelihood ratios (LR) were calculated with consideration of relevant factors. Of the 865 participants, 205 (23.7%) had ATB, including 58 (28.3%) microbiologically confirmed TB and 147 (71.7%) clinically diagnosed TB. 615 (71.7%) were non-TB. 45 (5.2%) cases were clinically indeterminate and excluded from the final analysis. In multivariate analysis, serous effusion was the only independent risk factor related to lower sensitivity (OR = 0.39, 95% CI: 0.18-0.81) among patients with ATB. Among non-TB patients, age, TB history, immunosuppressive agents/glucocorticoid treatment and lymphocyte count were the independent risk factors related to specificity of T-SPOT.TB. Sensitivity, specificity, PV+, PV-, LR+ and LR- of T-SPOT.TB for diagnosis of ATB were 78.5%, 74.1%, 50.3%, 91.2%, 3.0 and 0.3, respectively. This study suggests that influencing factors of sensitivity and specificity of T-SPOT.TB should be considered for interpretation of T-SPOT.TB results.

Utility of Th1-cell immune responses for distinguishing active tuberculosis from non-active tuberculosis: A case-control study.

Currently available Interferon-γ release assay (IGRA) cannot reliably differentiate active TB (ATB) from non-active TB (non-ATB). A study was performed to evaluate the value of Mycobacterium tuberculosis (MTB) specific Th1 cell immune responses which test IFN-γ and IL-2 simultaneous for differentiating ATB from non-ATB. Forty-nine newly diagnosed inpatients with ATB (26 pulmonary TB and 23 extrapulmonary TB) were enrolled as the ATB group. Forty-five volunteers with latent tuberculosis infection (LTBI) and twenty with evidence of previous TB were enrolled during the same period as the non-ATB group. Clinical examination and MTB specific Th1 cell immune responses were performed for all participants. After being stimulated with ESAT-6 and CFP-10, the median frequencies of single IL-2-, single IFN-γ-, and dual IFN-γ/IL-2-secreting T-cells were all higher in the ATB group than in the non-ATB group (20(8-45) vs. 7(3-13), P<0.001;131(44-308) vs. 10(6-27), P<0.001;25(9-74) vs. 7(3-23), P = 0.001, respectively). Evaluation of the diagnostic performance of detecting single IFN-γ-secreting T cells for pulmonary TB employed a cutoff value of 35 iSFCs/250,000 PBMC. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR), and negative likelihood ratio (NLR) were 92.3%, 80.0%, 64.9%, 96.3%, 4.62, and 0.10, respectively. For extrapulmonary TB, using a cutoff value of 23 iSFCs/ 250,000 PBMC, the sensitivity, specificity, PPV, NPV, PLR, and NLR were 91.3%, 76.9%, 58.3%, 96.2%, 3.96, and 0.11, respectively. When combining frequencies and proportion of single IFN-γ-secreting T cells, the test sensitivity was 100% in parallel tests and the specificity was 87.7% in serial tests for pulmonary TB. MTB specific Th1 cell immune responses (FluoroSpot) had value for the differentiation of ATB and non-ATB. Further confirmatory studies are indicated.