Cytokine and soluble adhesion molecule profiles and biomarkers for treatment monitoring in Re-treated smear-positive patients with pulmonary tuberculosis.

Relapse of pulmonary tuberculosis (PTB) is associated with a failure of the host immune system to control the invading Mycobacterium tuberculosis. Severe immunodeficiency or immune disorders may be the main reason for TB recurrence. This study aimed to quantify serum inflammatory cytokine and soluble adhesion molecule levels in Re-treated smear-positive PTB patients before and after re-anti-TB drug therapy. Serum samples were collected from 30 healthy controls and 215 Treated active PTB patients at baseline and 2, 4, and 6 months post-re-treatment. Levels of 18 serum cytokines and soluble adhesion molecules were measured by a high-throughput Cytometric Bead Array. At baseline, IL-1, IL-2, IL-12P70, and soluble CD62E levels were significantly higher in PTB patients than those in the healthy controls (p < 0.05); IL-4, IL-5, IL-7, IL-8, IL-10, IL-17, IL-21, soluble CD54, MIG, and TGF-ß levels in PTB patients were significantly lower than those in the healthy controls (p < 0.05), of which TGF-ß, IL-7, IL-8, IL-10, soluble CD54, and MIG were most notably (p < 0.0005). After re-treatment, IFN-γ, IL-2, IL-7, and soluble CD54 levels and IL-2/IL-10 and IFN-γ/IL-10 ratios showed an upward trend during the re-treatment period. They were more sensitive than other cytokines and adhesion molecules and could be effective as serum indicators for re-treatment response. The immune response was imbalance in treated smear-positive PTB patients: Th1 response was elevated, but Th2 and Th17 responses were reduced. Systematic and comprehensive understanding of the cytokine and soluble adhesion molecule profiles provides a theoretical basis for immuno-diagnosis, immunotherapy, and immuno-monitoring of Re-treated PTB patients.

The relationship between absolute counts of lymphocyte subsets and clinical features in patients with pulmonary tuberculosis.

BACKGROUND: The aim of the present study is to investigate the clinical value and characteristics of peripheral blood lymphocyte subsets in patients with pulmonary tuberculosis (PTB) using flow cytometry. METHODS: The absolute counts of T, CD4+ T, CD8+ T, natural killer (NK), NKT and B lymphocytes in 217 cases of PTB were detected, and the variations in lymphocyte subset counts between different ages and genders and between aetiological detection results and chest radiography results were analysed. RESULTS: In 75.3% of the patients with PTB, six subset counts were lower than the normal reference range, and 44% showed lower-than-normal CD4+ T lymphocyte levels. The counts of T, CD4+ T, CD8+ T and B lymphocytes were significantly lower in patients aged >60 years, and the NKT cell counts were significantly lower in female patients than in male patients. Among the patients with positive aetiological results, 40.8% had reduced CD8+ T counts; these were significantly lower than those in patients with negative aetiological results (P = 0.0295). The cell counts of T, CD4+ T, CD8+ T and B lymphocytes reduced as lesion lobe numbers increased. The counts of T, CD4+ T and CD8+ T lymphocytes were significantly higher in the group with lesions affecting one lobe than in the groups with two to three lobes or four to five lobes, and the counts of B lymphocytes were significantly higher in the group with one lobe and the group with two to three lobes than in the group with four to five lobes. The counts of CD4+ T and CD8+ T lymphocytes were highest in the no cavity group and showed a downward trend with the increase in cavities; the T lymphocyte count was significantly higher in the no cavity group than in the group with five or more cavities (P = 0.014), and the CD8+ T lymphocyte count was significantly higher in the no cavity group than in the group with one to two cavities and the group with five or more cavities (P = 0.001 and 0.01, respectively). CONCLUSIONS: In most patients with tuberculosis, immune function is impaired. The absolute counts of peripheral blood lymphocyte subsets are closely related to the aetiological results and lesion severity in patients with PTB; this could be used as evidence for immune intervention and monitoring curative effects.

[oil microbial biomass and enzyme activities among different artificial forests in Ziwuling, Northwest China.] / å­åˆå²­äººå·¥æž—åœŸå£¤å¾®ç”Ÿç‰©ç”Ÿç‰©é‡åŠé ¶æ´»æ€§.

Decades of ecological restoration on the Loess Plateau has achieved significant on-site benefits to reduce soil erosion and improve soil quality, with remarkable off-site effects of reducing sediment delivery to Yellow River. However, regional forest community succession is still far from being adequately developed. The Ziwuling forest region and its highly developed forest community, as an advanced eco-zone, can lend practical experience to other regions on the Loess Plateau and help to identify the most suitable tree species for a better regional restoration in the future. With the aim to systematically understand the potential effects of typical local tree species to soil properties, three typical and well-established artificial forests in the Ziwuling region, Robinia pseudoacacia, Pinus tabuliformis and Platycladus orientalis were investigated in this study, with the climax community Quercus wutaishanica as a reference. All the four forest type had comparable stand age (25 years). Soil samples from 0-20 cm layers were collected from those four plantations. The soil microbial biomass (carbon and nitrogen), soil enzyme (invertase, urease and alkaline phosphatase) activities and their correlations were measured and analyzed. The results showed that: 1) soil invertase activity ranged from 16.94 to 64.49 mg·g-1·24 h-1, the soil urease activity from 0.15 to 0.26 mg·g-1·24 h-1, and the alkaline phosphatase activity from 0.65 to 1.23 mg·g-1·24 h-1. The activities of those three enzymes were significantly higher in the P. orientalis soil that in the R. pseudoacacia and P. tabuliformis soils. The geometric average values in the P. orientalis soil were even greater than that in the Q. wutaishanica soil. 2) The soil microbial biomass carbon and nitrogen varied from 247.37 to 529.84 mg·kg-1 and 41.48 to 77.91 mg·kg-1, respectively. Both of them were significantly greater in the P. orientalis soil than that in the R. pseudoacacia and P. tabu-liformis soils. Even though the soil microbial biomass carbon in the P. orientalis soil remained lower than that in the Q. wutaishanica soil, its soil microbial biomass nitrogen was greater than in the Q. wutaishanica soil. 3) The dissolved organic carbon and nitrogen in the P. tabuliformis soil were much greater than that in other species, even greater than their own soil microbial biomass carbon and nitrogen. Such a result indicated that dissolved organic matter might play a more important role in providing plant available nutrients than microbial biomass in the P. tabuliformis soil. 4) The microbial biomass carbon and nitrogen were significantly positively correlated with the total organic carbon and the total nitrogen, particularly for the R. pseudoacacia and P. tabuliformis soils. There were significantly positive relationships between the soil invertase activity, urease activity and alkaline phosphatase activity, and their soil organic carbon, total nitrogen and total phosphorus contents. 5) Based on the results of principal component analysis, we concluded that the artificial forests types had obvious effects on soil microbial carbon and nitrogen, soil organic carbon, total nitrogen, the ratio of carbon to phosphorus, the ratio of nitrogen to phosphorus and urease activity. Overall, our findings suggested that P. orientalis is better than R. pseudoacacia and P. tabuliformis in term of improving soil properties in the south forest zone on the Chinese Loess Plateau.

[Effects of vegetation and topography features on ecological stoichiometry of soil and soil microbial biomass in the hilly-gully region of the Loess Plateau, China.] / 黄土丘陵区植被与地形特征对土壤和土壤微生物生物量生态化学计量特征的影响.

The objectives of this study were to explore the effects of vegetation type, topographic features and their combined effects on soil microbial biomass stoichiometry, so as to better understand the interaction of soil, soil microbes and nutrient cycling under different vegetation types in the hilly-gully region of the Loess Plateau. Soils from three vegetation zones (forest zone, forest-steppe and steppe) and five slope positions (south/north backslope, south/north shoulder and summit) were chosen and the effects of vegetation types and topography features on soil and C:N:P ratios in soil microbial biomass were investigated in this study. The results showed that, among the five slope positions, the highest concentrations of soil and soil microbial biomass C, N, P were found at the backslope position and the north-facing slope. The effects of vegetation types on soil and soil microbial biomass C, N and P in the two soil layers were significantly different, whereas the effects of slope aspect and positions were only numerically different. As for different soil layers, the topsoil (0-10 cm) was more affected by slope aspect, while the subsoil (10-20 cm) was more influenced by slop position. While the effects of vegetation type on soil C:N, C:P and N:P and soil microbial biomass C:N, C:P were significant, slope aspect and slope position only influenced soil C:P and N:P. Consequently, on the Loess Plateau, the effects of vegetation type on soil and soil microbial biomass C, N, P were stronger than those of the topographic features. The standardized major axis tests showed that C:N:P stoichiometry in soil microbes was well-constrained, especially in the steppe zone. The soil microbial biomass N:P might be used as a useful tool to assess nutrient limitation of ecosystem processes in terrestrial ecosystems. If combined with plant leaf N:P, they could provide more accurate information to estimate the nutrient limitation of fragile ecosystem in hilly-gully region of the Loess Plateau.

Diagnostic performance and problem analysis of commercial tuberculosis antibody detection kits in China.

BACKGROUND: The diagnosis of bacterium-negative pulmonary tuberculosis (TB) and extra-pulmonary TB is challenging clinically. The detection of the anti-TB antibody has an important, auxiliary, clinical diagnostic value. Therefore, TB antibody detection kits should be screened and evaluated, and the reagents with the highest sensitivity and specificity should be chosen and used clinically. METHODS: The diagnostic performance of 7 commercially available TB antibody detection kits (kits A, B, C, D, E, F and G) based on the gold immunoassay detection of immunoglobulin (Ig) G or IgM antibodies were simultaneously evaluated and compared in 62 TB cases and 56 non-TB cases in a laboratory. A retrospective analysis including 2549 cases was carried out to assess the clinical diagnosis values of bacteriological examinations and TB antibody tests (kits B and H used in the clinic). RESULTS: The sensitivities of TB antibody kits A, B, C, D, E, F and G in the sera from 62 TB patients were 50.0%, 83.9%, 38.7%, 9.7%, 48.4%, 69.4% and 79.0%, respectively; the sensitivities in the sera from 24 smear-negative TB patients were 29.2%, 79.2%, 29.2%, 12.5%, 29.2%, 54.2% and 79.2%, respectively; the specificities in the sera from 56 non-TB patients were 73.2%, 25.0%, 85.7%, 96.4%, 78.6%, 78.6% and 50.0%, respectively. Of the 2549 clinically diagnosed cases, there were 1752 pulmonary TB cases, 505 extra-pulmonary TB cases, 87 old pulmonary TB cases and 205 non-TB cases. The positive results for smear, culture, TB antibody kit B and kit H in pulmonary TB cases were 39.8% (543/1365), 48.6% (372/765), 45.8% (802/1752) and 25.2% (442/1752), respectively; the results in extra-pulmonary TB cases were 3.4% (6/178), 5.8% (4/69), 35.4% (179/505), and 11.3% (57/505), respectively; the results in old pulmonary TB cases were 0% (0/64), 0% (0/30), 32.2% (28/87), and 9.2% (8/87), respectively; and the results in non-TB cases were 0% (0/121), 0% (0/56), 21.5% (44/205), and 2.4% (5/205), respectively. Of 624 smear-positive and/or culture-positive pulmonary TB cases, the sensitivities of antibody test kits B and H were 53.0% and 36.4%, respectively. Of 901 smear-negative and/or culture-negative pulmonary TB cases, the sensitivities of antibody test kits B and H were 42.5% and 19.0%, respectively. The positive rate of antibody detection in the bacterium-positive pulmonary TB cases was significantly higher than that in the bacterium-negative pulmonary TB cases (P < 0.05). CONCLUSIONS: The colloidal gold-labeled TB antibody IgG detection assay is a simple, rapid and economical method that provides a better clinical auxiliary diagnosis value on TB, especially in smear-negative pulmonary TB and extra-pulmonary TB. The production, quality control, screening and evaluation of antibody detection kits are very important for its clinical application.

[Ecological stoichiometry characteristics of leaf-litter-soil in different plantations on the Loess Plateau, China]. / 黄土高原不同人工林叶片-凋落叶-土壤生态化学计量特征.

In order to research into the influence of Grain to Green Project in Ziwuling forest region, this paper took three typical plantations which were Robinia pseudoacacia plantation, Pinus tabuliformis plantation, and Platycladus orientalis plantation in the Ziwuling forest region of Shaanxi Province as research objects and analyzed the carbon, nitrogen and phosphorus contents of leaf, litter and soil among the three plantations. The results showed that the contents of C, N and P in the three plantations were in order of leaf > litter > soil, the contents of N and P in leaf of R. pseu-doacacia plantation were significantly higher than that of P. tabuliformis plantation and P. orientalis. Leaf N:P was 12.21, 5.36 and 6.09 in R. pseudoacacia plantation, P. tabuliformis plantation and P. orientalis plantation, respectively. It was indicated that the three species were all subject to N deficiency. C:N and C:P showed the trend of litter > leaf > soil, and N:P demonstrated the trend of leaf > litter > soil. There were highly significant positive relationships in C:N between leaf and litter in P. tabuliformis plantation. N and P in the leaf development process of R. pseu-doacacia plantation were proportionally absorbed, and proportionally remained in the litter after N and P resorption. R. pseudoacacia was the better plantation species than P. tabuliformis and P. orientalis in the south fo-rest zone on the Loess Plateau.

Potential novel markers to discriminate between active and latent tuberculosis infection in Chinese individuals.

Latent tuberculosis infection (LTBI) constitutes the main reservoir for reactivation tuberculosis. The finding of potential biomarkers for differentiating between TB and LTBI is very necessary. In this study, the immunological characteristics and potential diagnostic utility of Rv2029c, Rv2628 and Rv1813c proteins were assessed. These three proteins stimulated PBMCs from ELISPOT-positive LTBI subjects produced higher levels of IFN-γ in comparison with TB patients and ELISPOT-negative healthy subjects (p<0.05). BCG vaccination and non-TB respiratory disease had little influence on the immunological responses of Rv2029c and Rv2628 proteins (p>0.05). The LTBI diagnostic performance of Rv2029c was higher than Rv2628 and Rv1813c by ROC evaluation. But Rv2628 had much higher specificity than Rv2029c in active TB patients and uninfected healthy subjects. The IgG level against Rv1813c was higher in the TB group than in LTBI and uninfected healthy subjects (p<0.05). These results suggest that T cell response to Rv2628 and antibody against Rv1813c might be applicable as biomarkers to distinguish TB from LTBI and uninfected individuals.

[Screening of differentially expressed genes in colorectal cancer using human whole genomic oligonucleotide microarrays].

OBJECTIVE: To screen the differentially expressed genes in human colorectal cancer (CRC) tissue. METHODS: Affymetrix oligonucleotide microarrays HG-U133 representing 32,264 human genes including 19,308 known genes and 12,956 expressed sequence tags (ESTs) were used to detect the gene expressions of CRC tissue paired with normal mucosa tissue. The microarray findings were confirmed by real-time quantitative reverse transcriptase-polymerase chain reaction (FQ-PCR). The gene expression profiles were analyzed by intersection and complement, rank sum test and t test. RESULTS: Totally 3,125 genes and ESTs expressed differentially were detected in normal and cancer tissues, consisting of 974 up-regulated and 2,151 down-regulated genes with 247 ESTs present in CRC tissue and absent in normal mucosa and 162 ESTs absent in CRC tissue but present in normal mucosa. A percent of 80.1% of the differentially expressed genes were not reported in the literatures. CONCLUSION: The strategy of data mining provides a foundation for filtering molecular markers and interpreting molecular carcinogenesis of CRC.

Immune responses to latent tuberculosis antigen Rv2659c in Chinese populations.

BACKGROUND: The latency-associated antigen Rv2659c is a starvation-related protein of Mycobacterium tuberculosis (M. tuberculosis). It has potential use in tuberculosis (TB) control, but its immunological characteristics in Chinese populations are unclear. METHODS: In this study, immunological characteristics and potential diagnostic use of recombinant Rv2659c protein were assessed. Interferon-γ (IFN-γ) production from peripheral blood mononuclear cells (PBMC) was assayed by enzyme-linked immunospot (ELISPOT) in TB patients (80 cases), individuals who were purified protein derivative (PPD)-positive after Bacillus Calmette-Guérin (BCG) vaccination (27 cases), nontuberculous respiratory disease patients (30 cases), individuals who were identified by standard techniques as having latent TB infection (LTBI) (37 cases), and uninfected healthy individuals (75 cases). Serum immunoglobulin G (IgG) levels were assayed by enzyme-linked immunosorbent assay (ELISA) in TB patients (43 cases), LTBI individuals (36 cases) and uninfected healthy individuals (66 cases). RESULTS: When stimulated by rRv2659c, PBMC from LTBI individuals gave ELISPOT counts that were significantly higher than those from TB patients, BCG vaccinated individuals, non-TB respiratory disease patients and uninfected healthy individuals (p < 0.05). The rRv2659c stimulation gave detectable IFN-γ production in a higher proportion of persons with LTBI compared with TB patients and uninfected healthy individuals. BCG vaccination and non-TB respiratory disease had little influence on the PBMC response to rRv2659c. The levels of serum IgG specific for rRv2659c were not significantly different between LTBI individuals and TB patients (p > 0.05). CONCLUSION: These results suggest that rRv2659c has potential for the diagnosis of LTBI. This is the first clinical report of human immune recognition of Rv2659c in Chinese populations.