Clinical evaluation of an artificial intelligence-assisted cytological system among screening strategies for a cervical cancer high-risk population.

BACKGROUND: Primary cervical cancer screening and treating precancerous lesions are effective ways to prevent cervical cancer. However, the coverage rates of human papillomavirus (HPV) vaccines and routine screening are low in most developing countries and even some developed countries. This study aimed to explore the benefit of an artificial intelligence-assisted cytology (AI) system in a screening program for a cervical cancer high-risk population in China. METHODS: A total of 1231 liquid-based cytology (LBC) slides from women who underwent colposcopy at the Chinese PLA General Hospital from 2018 to 2020 were collected. All women had received a histological diagnosis based on the results of colposcopy and biopsy. The sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), false-positive rate (FPR), false-negative rate (FNR), overall accuracy (OA), positive likelihood ratio (PLR), negative likelihood ratio (NLR) and Youden index (YI) of the AI, LBC, HPV, LBC + HPV, AI + LBC, AI + HPV and HPV Seq LBC screening strategies at low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL) thresholds were calculated to assess their effectiveness. Receiver operating characteristic (ROC) curve analysis was conducted to assess the diagnostic values of the different screening strategies. RESULTS: The Se and Sp of the primary AI-alone strategy at the LSIL and HSIL thresholds were superior to those of the LBC + HPV cotesting strategy. Among the screening strategies, the YIs of the AI strategy at the LSIL + threshold and HSIL + threshold were the highest. At the HSIL + threshold, the AI strategy achieved the best result, with an AUC value of 0.621 (95% CI, 0.587-0.654), whereas HPV testing achieved the worst result, with an AUC value of 0.521 (95% CI, 0.484-0.559). Similarly, at the LSIL + threshold, the LBC-based strategy achieved the best result, with an AUC of 0.637 (95% CI, 0.606-0.668), whereas HPV testing achieved the worst result, with an AUC of 0.524 (95% CI, 0.491-0.557). Moreover, the AUCs of the AI and LBC strategies at this threshold were similar (0.631 and 0.637, respectively). CONCLUSIONS: These results confirmed that AI-only screening was the most authoritative method for diagnosing HSILs and LSILs, improving the accuracy of colposcopy diagnosis, and was more beneficial for patients than traditional LBC + HPV cotesting.

Up-regulated SAMD9L modulated by TLR2 and HIF-1α as a promising biomarker in tuberculosis.

The aim of this study was to identify potential biomarkers of TB in blood and determine their function in Mtb-infected macrophages. First of all, WGCNA was used to analyse 9451 genes with significant changes in TB patients' whole blood. The 220 interferon-γ-related genes were identified, and then 30 key genes were screened using Cytoscape. Then, the AUC values of key genes were calculated to further narrow the gene range. Finally, we identified 9 genes from GSE19444. ROC analysis showed that SAMD9L, among 9 genes, had a high diagnostic value (AUC = 0.925) and a differential diagnostic value (AUC>0.865). To further narrow down the range of DEGs, the top 10 hub-connecting genes were screened from monocytes (GSE19443). Finally, we obtained 4 genes (SAMD9L, GBP1, GBP5 and STAT1) by intersections of genes from monocytes and whole blood. Among them, it was found that the function of SAMD9L was unknown after data review, so this paper studied this gene. Our results showed that SAMD9L is up-regulated and suppresses cell necrosis, and might be regulated by TLR2 and HIF-1α during Mtb infection. In addition, miR-181b-5p is significantly up-regulated in the peripheral blood plasma of tuberculosis patients, which has a high diagnostic value (AUC = 0.969).

Zearalenone Promotes LPS-Induced Oxidative Stress, Endoplasmic Reticulum Stress, and Accelerates Bovine Mammary Epithelial Cell Apoptosis.

Both zearalenone (ZEA) and lipopolysaccharide (LPS) can induce oxidative stress, and even apoptosis in bovine mammary epithelial cells (MAC-T), but not much attention has been given to the synergistic effect of ZEA and LPS. In this study, we treated MAC-T cells with different concentrations of LPS (1, 10, 50, and 100 µg/mL) and ZEA (5, 15, and 30 µM) to induce cell damage. Previous results show that MAC-T cell viability decreases with increasing LPS concentration. Meanwhile, 1 µg/mL LPS and ZEA were selected for combined treatment in subsequent studies. It was found that co-treatment with ZEA and LPS increases the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), decreases mitochondrial membrane potential (MMP), and superoxide dismutase (SOD), and reduces glutathione (GSH). ZEA and LPS are found to activate endoplasmic reticulum (ER) stress by increasing the expression of glucose-regulated protein 78 kDa (GRP78), activating transcription factor 6 (ATF6) and C/EBP homologous protein (CHOP). It increases cell apoptosis by suppressing the expression of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2), indicated by up-regulation of Bcl2-associated X protein (Bax) and Cysteinyl aspartate-specific proteinases 3 (caspase-3) expression. The above results suggest that the synergistic effect of ZEA and LPS aggravate cytotoxicity.

Increased UBE2L6 regulated by type 1 interferon as potential marker in TB.

The aim of this study is to identify potential biomarker of tuberculosis (TB) and determine its function. Differentially expressed mRNAs(DEGs) were selected from a blood database GSE101805, and then, 30 key genes were screened using STING, Cytoscape and further functionally enriched. We then found that only 6 of 13 genes related to ubiquitination (the first in the functional enrichment) were increased significantly. ROC analysis showed that UBE2L6, among 6 genes, had the highest diagnostic value, and then, we found that it also had mild value in differential diagnosis. Moreover, our analysis showed that UBE2L6 may be upregulated by type I interferon, which was further confirmed by us. In addition, we also found that UBE2L6 inhibits the apoptosis of Mycobacterium tuberculosis(Mtb)infected macrophages. Subsequently, we discovered that miR-146a-5p, which may target UBE2L6, is reduced in peripheral blood mononuclear cells (PBMC) and plasma of TB, and it also had certain diagnostic efficiency(AUC=0.791). In brief, we demonstrated that UBE2L6 as well as miR-146a-5p is a potential biomarker for TB and UBE2L6,which may also plays important role in TB by, at least, modulating Mtb-infected macrophage apoptosis.

STAT1 and its related molecules as potential biomarkers in Mycobacterium tuberculosis infection.

Tuberculosis (TB) is a severe infectious disease that seriously endangers human health. The immune defence mechanism of the body against TB is still unclear. The purpose of this study was to find the key molecules involved in the immune defence response during TB infection, and provide reference for the treatment of TB and further understanding of the immune defence mechanism of the body. Data from GSE83456 were downloaded from GEO data sets for analysis, and a total of 192 differentially expressed genes were screened out. Most of these genes are enriched in the interferon signalling pathway and are defence response-related. We also found that STAT1 plays an important role in the immune defence of TB infection and it is one of the key genes related to interferon signalling pathway. STAT1-related molecules including hsa-miR-448, hsa-miR-223-3p, SAMD8_hsa_circRNA 994 and TWF1_hsa_circRNA 9897 were therefore screened out. Furthermore, expression levels of hsa-miR-448 and hsa-miR-223-3p were then verified by qRT-PCR. Results showed that both hsa-miR-448 and hsa-miR-223-3p were down-regulated in plasma from patients with pulmonary TB. Taken together, our data indicate that an mRNA-miRNA-circRNA interaction chain may play an important role in the infection of MTB, and STAT1 and related molecules including hsa-miR-223-3p, has-miR-448, SAMD8_hsa_circRNA994 and TWF1_hsa_circRNA9897 were identified as potential biomarkers in the development of active TB.

miR-18a promotes Mycobacterial survival in macrophages via inhibiting autophagy by down-regulation of ATM.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is one of leading causes of global deaths. This study aimed to explore the role of miR-18a in RAW264.7 cells response to Mtb infection. Exosomes derived from Mtb-infected cells were isolated and further validated by size, transmission electron microscopy and Western blot. RT-PCR was utilized to measure miR-18a expression. Cell viability and ultrastructure were examined by CFU counting, CCK-8 and electron microscope, respectively. Potential target genes of miR-18a were predicted with bioinformatics and further confirmed using RT-PCR, Western blot and laser confocal microscope analysis, respectively. LC3, AMPK and mTOR were measured using Western blot. We found that miR-18a was induced both in Mtb-infected RAW264.7 cells and its derived exosomes compared with the controls. In addition, up-regulation of miR-18a promoted intracellular Mtb survival, attenuated cell viability and reduced LC3-II level, while its down-regulation had the opposite effect. miR-18a overexpression suppressed level of ATM, one possible target of miR-18a, while its underexpression enhanced ATM. We also found that inhibition of ATM induced LC3-II decrease in Mtb-infected cells and could reverse the increase of LC3-II caused by inhibition of miR-18a. Moreover, down-regulation of miR-18a increased p-AMPK level while reduction of ATM could reverse the change. Taken together, our results suggest that miR-18a is up-regulated in macrophages response to Mtb infection, and it promotes intracellular Mtb survival through repressing autophagic process by down-regulation of ATM pathway. This provides new thought for TB pathogenesis, diagnosis and treatment.

Study on amniotic fluid metabolism in the second trimester of Trisomy 21.

BACKGROUND: Trisomy 21 is a common aneuploid condition in humans and accounts for approximately one quarter of all aneuploid live births. To date, early diagnosis of Trisomy 21 remains a challenging task. Metabolomics may prove an innovative tool to study the early pathophysiology of Trisomy 21 at a functional level. METHODS: Ultra-performance liquid chromatography coupled with mass spectrometer (UPLC-MS) was used for untargeted metabolomic analysis of amniotic fluid samples from women having normal and trisomy 21 fetuses. RESULTS: Many significantly changed metabolites were identified between amniotic fluid samples from Trisomy 21 pregnancies and normal euploid pregnancies, such as generally lower levels of several steroid hormones and their derivatives, higher levels of glutathione catabolites coupled with lower levels of gamma-glutamyl amino acids, and increased levels of phospholipid catabolites, sugars, and dicarboxylic acids. The identification of a human milk oligosaccharide in amniotic fluid may worth further investigation, since confirmation of this observation may have significant implications for regulation of fetal development. CONCLUSIONS: The metabolisms in amniotic fluid from Trisomy 21 and normal pregnancies are quite different, and some of the significantly changed metabolites may be considered as candidates of early diagnostic biomarkers for Trisomy 21.

Effects of dietary grape pomace on the intestinal microbiota and growth performance of weaned piglets.

Grape pomace (GP) is an abundant by-product from wine production and is rich in phenolic compounds, unsaturated fatty acids, dietary fibre and beneficial bacteria. In this study, weaned piglets were fed a basic diet supplemented with 5% GP for 4 weeks. Compared with those in the control (CON) group, it was found that the proportion of Lactobacillus delbrueckii, Olsenella umbonata and Selenomonas bovis in the caecum and the villus height and villus height/crypt depth ratio (VCR) of the jejunum were both significantly increased in the GP group (p < 0.05). Meanwhile, at the mRNA expression level, several proinflammatory cytokines (IL-1ß, IL-8, IL-6 and TNF-α) were significantly downregulated (p < 0.05) in piglet caecal tissue, and the short-chain fatty acid receptors (GPR41 and GPR43) were not significantly upregulated. In contrast, the levels of IgG was significantly increased (p < 0.05) in the sera of weaned piglets in the GP group. However, no difference in growth performance between the two groups of piglets was detected. These results show that GP had no adverse effects on the growth performance of piglets, but GP can promote the content of some beneficial bacteria in the caecum; this effect is conducive to improving the disease resistance potential of piglets.

Signature of circular RNAs in peripheral blood mononuclear cells from patients with active tuberculosis.

The study was to characterize the expression profiles of circular RNAs (circRNAs) in peripheral blood mononuclear cells (PBMCs) from active tuberculosis (TB) patients and to investigate their function. Microarray was applied to detect circRNA expression and reverse transcription-quantitative polymerase chain reaction was conducted to validate the microarray results. Meanwhile, receiver operating characteristic curve (ROC) curve was calculated to evaluate the predictive power of the selected circRNAs for TB diagnosis. Additionally, circRNA/miRNA interaction was predicted based on miRNA target prediction software, and gene ontology as well as Kyoto Encyclopedia of Genes and Genomes pathway analysis were used to predict their biological function. In total, 171 circRNAs were found to be dysregulated in TB samples. Specifically, circRNA_103017, circRNA_059914 and circRNA_101128 were confirmed to be increased, while circRNA_062400 was decreased in TB samples. ROC analysis revealed that circRNA_103017 had potential value for TB diagnosis, followed by circRNA_059914 and circRNA_101128. Moreover, circRNA_101128 expression in TB samples was negatively correlated with the level of its possible target let-7a and bioinformatics analysis showed that circRNA_101128 was potentially involved in MAPK and P13K-Akt pathway possibly via modulation of let-7a. Taken together, our results indicated that some dysregulated circRNAs were potential biomarkers for the diagnosis of TB and circRNA_101128-let-7a interplay may play considerable role in PBMCs response to Mtb infection.

Downregulation of miR-20b-5p facilitates Mycobacterium tuberculosis survival in RAW 264.7 macrophages via attenuating the cell apoptosis by Mcl-1 upregulation.

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb). The interaction between Mtb and macrophages, which is regulated by microRNAs, determines the development of TB. However, the function of microRNA-20b-5p (miR-20b-5p) in RAW 264.7 macrophages against Mtb remains unknown. In this study, we analyzed the expression level of miR-20b-5p in macrophage responses to Mtb infection and exosomes derived from macrophages after Mtb infection. MiR-20b-5p mimics and inhibitor were, respectively, transfected to evaluate the effect of miR-20b-5p on Mtb and macrophages. In addition, the targets of miR-20b-5p were predicted by a bioinformatics analysis. The macrophages were respectively transfected with miR-20b-5p mimics and inhibitor to determine the messenger RNA expression levels of the targets by reverse transcription-polymerase chain reaction assay. The results revealed that the miR-20b-5p expression level was decreased in the infected macrophages at different times. MiR-20b-5p was shown in the exosomes released from macrophages infected with Mtb. Upregulation of the miR-20b-5p level suppressed the survival of Mtb in macrophages, while downregulation of the miR-20b-5p level enhanced the survival of Mtb in macrophages. Overexpression of miR-20b-5p decreased the cell viability and induced apoptosis in Mtb-infected macrophages, while underexpression of miR-20b-5p increased the cell vitality and attenuated apoptosis in Mtb-infected macrophages. The bioinformatics analysis revealed that Mcl-1 was a target of miR-20b-5p. MiR-20b-5p negatively regulated the expression of Mcl-1. Overall, this study is the first to demonstrate the effect of miR-20b-5p on Mtb infection and present miR-20b-5p and exosomes as the potential therapeutic targets of TB.

Zearalenone induces apoptosis in bovine mammary epithelial cells by activating endoplasmic reticulum stress.

Zearalenone (ZEA) is a common mycotoxin produced by fungi within the genus Fusarium. However, few studies have examined the direct effects of the toxin on the mammary glands. In the present study, the effects of ZEA treatment on bovine mammary epithelial cells (MAC-T) from dairy cows were investigated. The cells were treated with different concentrations of ZEA to evaluate the effect of the toxin on cell viability, intracellular reactive oxygen species (ROS) concentrations, mitochondrial membrane potential, endoplasmic reticulum (ER) stress, and the expression of apoptosis-related genes. The results indicated that different concentrations (5, 10, 15, 20, 25, 30, 50, 60, or 100 µM) of ZEA were able to inhibit growth of MAC-T cells. After exposing the MAC-T cells to 30 µM ZEA, compared with the control group, ROS levels increased, mitochondrial membrane potential decreased, and mRNA expression of the ER-specific stress-related genes GRP78, HSP70, ATF6, EIF2A, ASK1, and CHOP was upregulated in the ZEA-treated group. Further, we analyzed the increase in apoptotic rate by flow cytometry. At the mRNA level, compared with the control group, the expression of the apoptosis-promoting gene BAX was increased in the ZEA-treated group, the expression of the inhibitory gene BCL2 decreased, and the expression of the gene CASP3 increased. We observed a significant increase in caspase-3 activity in ZEA-treated MAC-T cells. Furthermore, the apoptotic rate of the cells in the ZEA group treated with 4-phenylbutyric acid (ER stress inhibitor) decreased and the mRNA expression levels of ER stress markers GRP78 and CHOP decreased. Compared with the ZEA treatment group, the mRNA expression level of the apoptosis-related gene BAX was decreased and the expression level of BCL2 was increased in the ZEA + 4-phenylbutyric acid cotreatment group. These findings indicate that ZEA-induced ER stress increases apoptosis in MAC-T cells. The treatment of MAC-T cells with ZEA reduced cell viability, increased ROS content, decreased mitochondrial membrane potential, increased ER stress marker expression, and induced apoptosis.

The Immune Escape Mechanisms of Mycobacterium Tuberculosis.

Epidemiological data from the Center of Disease Control (CDC) and the World Health Organization (WHO) statistics in 2017 show that 10.0 million people around the world became sick with tuberculosis. Mycobacterium tuberculosis (MTB) is an intracellular parasite that mainly attacks macrophages and inhibits their apoptosis. It can become a long-term infection in humans, causing a series of pathological changes and clinical manifestations. In this review, we summarize innate immunity including the inhibition of antioxidants, the maturation and acidification of phagolysosomes and especially the apoptosis and autophagy of macrophages. Besides, we also elaborate on the adaptive immune response and the formation of granulomas. A thorough understanding of these escape mechanisms is of major importance for the prevention, diagnosis and treatment of tuberculosis.

Deoxynivalenol induces oxidative stress, inflammatory response and apoptosis in bovine mammary epithelial cells.

Deoxynivalenol (DON) is a toxic secondary metabolite produced by Fusarium graminearum. It is one of the most common feed contaminants that poses a serious threat to the health and performance of dairy cows. This study investigated the in vitro cytotoxicity of DON on bovine mammary epithelial cells (MAC-T). DON at different concentrations (0.25, 0.3, 0.5, 0.8, 1 or 2 µg/ml) inhibited the growth of MAC-T cells after 24 hr of exposure (p < .001). DON at 0.25 µg/ml increased lactate dehydrogenase (LDH) leakage (p < .05); decreased glutathione (GSH) levels (p < .001), total superoxide dismutase (T-SOD) activity and total antioxidant capacity (T-AOC; p < .01); and increased malondialdehyde (MDA) concentration (p < .01) in MAC-T cells after 24 hr of exposure. We also observed that DON increased reactive oxygen species (ROS) levels in cells incubated for 9, 15 and 24 hr (p < .001). DON at 0.25 µg/ml triggered oxidative damage in MAC-T cells. Furthermore, it induced an inflammatory response in the cells incubated for 9, 15 and 24 hr (p < .05) by increasing the mRNA expression levels of nuclear factor kappa B, myeloid differentiation factor 88 (MyD88), tumour necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6, cyclooxygenase-2 and IL-8. We further examined the effect of DON on apoptosis. DON prevented normal proliferation of MAC-T cells by blocked cell cycle progression in 24 hr (p < .001). In addition, the apoptosis rate measured using annexin V-FITC significantly increased (p < .05) with increase in the mRNA expression level of Bax (p < .01) and increase in the Bax/Bcl-2 ratio (p < .01) in cells incubated for 24 hr. In summary, DON exerts toxic effects in MAC-T cells by causing oxidative stress, inducing an inflammatory response, affecting cell cycle and leading to apoptosis.

Dysregulated circRNAs in plasma from active tuberculosis patients.

Endogenous circular RNAs (circRNAs) have been reported in various diseases. However, their role in active TB remains unknown. The study was aimed to determine plasma circRNA expression profile to characterize potential biomarker and improve our understanding of active TB pathogenesis. CircRNA expression profiles were screened by circRNA microarrays in active TB plasma samples. Dysregulated circRNAs were then verified by qRT-PCR. CircRNA targets were predicted based on analysis of circRNA-miRNA-mRNA interaction. GO and KEGG pathway analyses were used to predict the function of circRNA. ROC curve was calculated to evaluate diagnostic value for active TB. A total of 75 circRNAs were significantly dysregulated in active TB plasma. By further validation, hsa_circRNA_103571 exhibited significant decrease in active TB patients and showed potential interaction with active TB-related miRNAs such as miR-29a and miR-16. Bioinformatics analysis revealed that hsa_circRNA_103571 was primarily involved in ras signalling pathway, regulation of actin cytoskeleton, T- and B-cell receptor signalling pathway. ROC curve analysis suggested that hsa_circRNA_103571 had significant value for active TB diagnosis. Circulating circRNA dysregulation may play a role in active TB pathogenesis. Hsa_circRNA_103571 may be served as a potential biomarker for active TB diagnosis, and hsa_circRNA_103571-miRNA-mRNA interaction may provide some novel mechanism for active TB.

Activation of TLR7 Inhibition of Mycobacterium Tuberculosis Survival by Autophagy in RAW 264.7 Macrophages.

The aim of the study was to evaluate the effect of regulation of TLR7 on Mycobacterium tuberculosis (Mtb) survival in macrophages. TLR7 expression in macrophages infected by Mtb was detected by RT-PCR and Western blotting. Regulation of TLR7 was achieved by single strand RNA (ssRNA) or siRNA. The effects of TLR7 on Mtb survival and cell viability were detected by acid fast staining and cell counting kit-8, respectively. Cell ultrastructure was observed via transmission electron microscopy (TEM), and autophagy related protein LC3 was analyzed by Western blotting. TLR7 in Mtb infected macrophages was up-regulated and up-regulation of TLR7 could eliminate intracellular Mtb. Up-regulation of TLR7 could increase viability of Mtb infected cells, while down-regulation of TLR7 induced decrease of cell viability compared with the controls. Autophagosome was significantly increased in the Mtb infected macrophages after up-regulation of TLR7 and LC3-II protein showed obvious increase compared with the controls. Autophagosome could not be detected in macrophages after down-regulation of TLR7, rough endoplasmic reticulum was dilated, and nuclear week gap was widened. Moreover, LC3-II protein was reduced in Mtb infected macrophages based upon the down-regulation of TLR7. Up-regulation of TLR7 could eliminate intracellular Mtb through autophagy. J. Cell. Biochem. 118: 4222-4229, 2017. © 2017 Wiley Periodicals, Inc.

Aberrantly Expressed Long Non-Coding RNAs In CD8+ T Cells Response to Active Tuberculosis.

Dysregulated expression of long noncoding RNAs (lncRNAs) has been demonstrated as being implicated in a variety of human diseases. In the study we aimed to determine lncRNA profile in CD8+ T cells response to active tuberculosis (TB). We examined the lncRNA expression by microarray in circulating CD8+ T cells isolated from patients with active TB and healthy controls. Change predictions to analysis was used to address functional roles of the deregulated mRNAs. Real-time quantitative PCR (RT-qPCR) was used to validate the microarray result. In total, 328 lncRNAs and 356 mRNAs were differentially expressed in TB CD8+ T cells. Upregulated mRNAs were mainly enriched in cAMP signaling pathway, calcium signaling pathway, and TGF-beta signaling pathway, while downregulated mRNAs were enriched in antigen processing and presentation and natural killer cell mediated cytotoxicity in TB CD8+ T cells. Interestingly, we found that heme oxygenase 1 (HMOX1) was decreased in active TB CD8+ T cells, while its nearby lincRNA XLOC_014219 was upregulated. Subsequent RT-qPCR results confirmed the changes. This is the first research addressing lncRNA expression profiles in active TB CD8+ T cells. The aberrantly expressed lncRNAs observed in the study may provide clues to the dysfunction of CD8+ T cells and so to the pathophysiological properties of active TB. Further studies should focus on the function of lncRNAs involved in active TB. J. Cell. Biochem. 118: 4275-4284, 2017. © 2017 Wiley Periodicals, Inc.

Deregulated microRNAs in CD4+ T cells from individuals with latent tuberculosis versus active tuberculosis.

The mechanisms of latent tuberculosis (TB) infection remain elusive. Roles of microRNA (miRNA) have been highlighted in pathogen-host interactions recently. To identify miRNAs involved in the immune response to TB, expression profiles of miRNAs in CD4(+) T cells from patients with latent TB, active TB and healthy controls were investigated by microarray assay and validated by RT-qPCR. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to analyse the significant functions and involvement in signalling pathways of the differentially expressed miRNAs. To identify potential target genes for miR-29, interferon-γ (IFN-γ) mRNA expression was measured by RT-qPCR. Our results showed that 27 miRNAs were deregulated among the three groups. RT-qPCR results were generally consistent with the microarray data. We observed an inverse correlation between miR-29 level and IFN-γ mRNA expression in CD4(+) T cells. GO and KEGG pathway analysis showed that the possible target genes of deregulated miRNAs were significantly enriched in mitogen-activated protein kinase signalling pathway, focal adhesion and extracellular matrix receptor interaction, which might be involved in the transition from latent to active TB. In all, for the first time, our study revealed that some miRNAs in CD4(+) T cells were altered in latent and active TB. Function and pathway analysis highlighted the possible involvement of miRNA-deregulated mRNAs in TB. The study might help to improve understanding of the relationship between miRNAs in CD4(+) T cells and TB, and laid an important foundation for further identification of the underlying mechanisms of latent TB infection and its reactivation.

[Expression and bioinformatic analysis of miR-29 family, target gene IFN-γ in CD4(+) T cells from subjects with latent tuberculosis infection].

OBJECTIVE: To investigate the effect of latent and active pulmonary tuberculosis(TB) on expression of miR-29 family and target gene IFN-γ in CD4(+)T cells. METHODS: Subjects from two hospitals of Weifang were enrolled from March 2012 to December 2012 and divided into three groups: active TB group(30 cases), latent tuberculosis infection(LTBI) group(25 cases) and healthy control group(30 cases). CD4(+) T cells in blood were collected from the three groups.Levels of miR-29a, miR-29b and miR-29c were measured by nucleic acid hybridization and RT-qPCR.Expression of IFN-γ was analyzed by RT-qPCR. Target genes of miR-29 family were predicted with both TargetScan and PicTar.GO annotation and pathway overrepresentation were further analyzed with David database and Cytoscape. RESULTS: Levels of miR-29a, miR-29b and miR-29c showed significant differences among the three groups(P < 0.05): levels of miR-29b and miR-29c in the active TB group(561.63 ± 65.36, 281.85 ± 42.78) were higher than the healthy controls(260.74 ± 38.69, 128.21 ± 19.98), but lower than the LTBI group(2030.29 ± 321.68, 620.93 ± 79.14); expression of miR-29a in the healthy control group(913.95 ± 104.73) were higher than the active TB group(323.37 ± 54.38), but lower than the LTBI group(4782.13 ± 567.81).Level of IFN-γ showed significant differences among the three groups(P < 0.05): level of IFN-γ in the LTBI group(0.45 ± 0.09) were lower than the healthy controls(1.00), but higher than the active TB group(0.11 ± 0.03). The target genes of miR-29 family mainly existed in molecular function such as extracellular matrix structural constituent and transcription regulator activity.In KEGG pathway, the gene set mostly existed in signaling pathway such as Focal adhesion,ECM-receptor interaction and mTOR signaling pathway. CONCLUSION: The expression of miR-29 family was increased and target gene IFN-γ in CD4(+) T cells was decreased by latent and active pulmonary TB, which might play important role in alteration of signal pathway.

[Research advance on bacteriophage therapy in bacterial infection].

Bacteriophage is a bacterium dependent virus. It has unique advantages in the treatment of bacterial infection, especially infection caused by drug-resistant bacteria. Its metabolic kinetics and route of administration are the current research focus. Bacteriophage lytic enzyme, as a new therapeutic method, has more advantages than active bacteriophage. This review is focused on the recent progress in bacteriophage research, including the mechanism of bacteria lysis, the route of administration, the application of genetic engineering, etc.

[Expression of miR-29a in serum of patients with pulmonary tuberculosis and prediction of its function with bioinformatics analysis].

OBJECTIVE: To analyze the expression of miR-29a in serum of patients with pulmonary tuberculosis, and to predict and analyze function of its target genes for further studying of its biological function and regulatory mechanism in tuberculosis (TB). METHODS: Fasting venous blood samples were collected from 65 untreated patients with active pulmonary TB (case group) and 45 healthy controls (control group) in the morning, respectively, and then sera were isolated. Total RNA was extracted with TRIzol reagent and was further purified. RNA quality was measured by gel electrophoresis and ultraviolet spectrophotometer. Nucleic acid hybridization and real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were performed to investigate expression of miR-29a in serum. Both TargetSean and PicTar software were used to predict comprehensively target genes of miR-29a and their intersection was regarded as target genes of miR-29a. Gene ontology of target genes was analyzed with DAVID database and three category annotations were extracted, respectively. GO overrepresentation was further analyzed by BINGO of Cytoscape. Enriched pathways of target genes were analyzed. RESULTS: The hybridization result showed that miR-29a was increased in serum of the case group (854 ± 93) compared to the control group (80 ± 22) (t = 3.541, P < 0.05). The PCR result showed that compared to the control group (0.18 ± 0.07), miR-29a was increased in serum of the case group (1.35 ± 0.62) (t = 2.987, P < 0.05). The target genes were mainly involved in biological processes including regulation of transcription, molecular function including metal ion binding, and cellular components including extracellular matrix, respectively. In the KEGG pathway, the target gene set was significantly enriched in the 7 signaling pathways including adhesion, ECM-receptor interaction and so on. In the PANTHER pathway, the gene set mostly existed in integrin signaling pathway, cadherin signaling pathway and Wnt signaling pathway. CONCLUSIONS: Level of miR-29a was increased significantly in serum of patients with active pulmonary tuberculosis. Target genes of miR-29a were mainly involved in biological processes including cell adhesion, regulation of transcription and so on.