The changes of intestinal flora and its relevance with memory Tfh and B cells in rheumatoid arthritis patients treated with csDMARDs and csDMARDs + bDMARDs.

BACKGROUND: A growing body of experimental and clinical evidence has implicated gut microbiota in the onset and course of rheumatoid arthritis (RA). The imbalance of intestinal flora in RA patients may lead to abnormal expression of immune cells and related cytokines. PURPOSE: Conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) and conventional synthetic disease-modifying antirheumatic drugs combined with biological disease-modifying antirheumatic drugs (csDMARDs + bDMARDs) are widely used to treat RA, but the characteristics of gut microbiota before and after treatment and their relationship with memory Tfh/B cells and cytokines remain unclear. METHODS: Stool samples were collected from 50 RA patients and 25 healthy controls (HCs) for 16SrRNA gene sequencing. We examined the proportion of lymphocyte subsets in healthy controls and RA patients. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of related cytokines in serum. The α and ß diversity of intestinal flora, and the correlation between intestinal flora and clinical indicators, lymphocyte subsets, cytokines were analyzed. RESULT: At the genus level, Ruminococcaceae_Ruminococcus was decreased in the csDMARDs and csDMARDs + bDMARDs treatment group, whereas Faecalibacterium was reduced in the csDMARDs treatment group, compared to untreated group. CD4+CD45RO+CCR7+CXCR5+central memory Tfh cells and CD4+CD45RO+CCR7-CXCR5+effector memory Tfh cells were significantly lower in the csDMARDs + bDMARDs treatment group than in untreated group. CD19+CD27+IgD+pre-switched memory B cells were higher in the csDMARDs and csDMARDs + bDMARDs treatment groups, whereas CD19+CD27+IgD-switched memory B cells were significantly lower than in untreated group. Ruminococcaceae_Ruminococcus was negatively correlated with CD19+CD27+IgD+ pre-switched memory B cells but positively correlated with CD4+CD45RO+CCR7-CXCR5+effector memory Tfh and CD19+CD27+IgD-switched memory B cells in patients with RA treated with DMARDs. CONCLUSION: The gut microbiota, memory Tfh cells, memory B cells, and cytokines of patients with RA changed significantly under different treatment regimens and had certain correlations with the clinical indicators of RA.

Radiosynthesis of a novel antisense imaging probe targeting LncRNA HOTAIR in malignant glioma.

BACKGROUND: Long non-coding RNA (LncRNA) HOTAIR was amplified and overexpressed in many human carcinomas, which could serve as a useful target for cancer early detection and treatment. The 99mTc radiolabeled antisense oligonucleotides (ASON) could visualize the expression of HOTAIR and provide a diagnostic value for malignant tumors. The aim of this study was to evaluate whether liposome-coated antisense oligonucleotide probe 99mTc-HYNIC-ASON targeting HOTAIR can be used in in vivo imaging of HOTAIR in malignant glioma xenografts. METHODS: The ASON targeting LncRNA HOTAIR as well as mismatched ASON (ASONM) were designed and modified. The radiolabeling of 99mTc with two probes were via the conjugation of bifunctional chelator HYNIC. Then probes were purified by Sephadex G25 and tested for their radiolabeling efficiency and purity, as well as stability by ITLC (Instant thin-layer chromatography) and gel electrophoresis. Then the radiolabeled probes were transfected with lipofectamine 2000 for cellular uptake test and the next experimental use. Furthermore, biodistribution study and SPECT imaging were performed at different times after liposome-coated 99mTc-HYNIC-ASON/ASONM were intravenously injected in glioma tumor-bearing mice models. All data were analyzed by statistical software. RESULTS: The labeling efficiencies of 99mTc-HYNIC-ASON and 99mTc-HYNIC-ASONM measured by ITLC were (91 ± 1.5) % and (90 ± 0.6) %, respectively, and both radiochemical purities were more than 89%. Two probes showed good stability within 12 h. Gel electrophoresis confirmed that the oligomers were successfully radiolabeled no significant degradation were found. Biodistribution study demonstrated that liposome-coated antisense probes were excreted mainly through the kidney and bladder and has higher uptake in the tumor. Meanwhile, the tumor was clearly shown after injection of liposome coated 99mTc-HYNIC-ASON, and its T/M ratio was higher than that in the non-transfection group and mismatched group. No tumor was seen in mismatched and blocking group. CONCLUSION: The liposome encapsulated 99mTc-HYNIC-ASON probe can be used in the in vivo, real-time imaging of LncRNA HOTAIR expression in malignant glioma.

Intravoxel incoherent motion diffusion-weighted magnetic resonance imaging in characterization of axillary lymph nodes: Preliminary animal experience.

PURPOSE: To investigate the diagnostic value of intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) for discriminating axillary metastatic from non-metastatic lymph nodes (LNs) in rabbit models. MATERIALS AND METHODS: The institutional animal care and use committee approved this study. Forty New Zealand white rabbits were randomly divided into two groups. The axillary LN models were created by inoculating VX2 cell suspension and complete Freund's adjuvant in the mammary glands of 20 female rabbits of each group, respectively. Conventional MRI and IVIM DWI were performed after animal models successfully established. Images of axillary LNs were analyzed with regard to long-axis diameter (L), short-axis diameter (S), apparent diffusion coefficient (ADC) and IVIM parameters (D, D*, f). Receiver operating characteristic analyses were conducted to determine the diagnostic performance of aforementioned criteria. RESULTS: A total of 42 metastatic and 30 non-metastatic LNs were successfully isolated. ADC and D of metastatic LNs were significantly lower than those of non-metastatic ones (all P < 0.001), whereas D* was statistically higher (P = 0.033). L, S, and f showed no significant difference between the two groups (P = 0.089, 0.058, 0.054, respectively). Optimal cutoff values, area under the curve, sensitivity, and specificity for differentiation were as follows: ADC = 1.101 × 10-3 mm2/s, 0.886, 78.6%, 90.0%; D = 0.938 × 10-3 mm2/s, 0.927, 83.3%, 93.3%; and D* = 12.635 × 10-3 mm2/s, 0.657, 52.4%, 80.0%. CONCLUSION: IVIM DWI is useful to distinguish metastatic from non-metastatic LNs in axilla. D was the most discriminative variable for predicting metastatic LNs.

Correlations between the minimum and mean apparent diffusion coefficient values of hepatocellular carcinoma and tumor grade.

PURPOSE: To investigate the correlations between the minimum and mean apparent diffusion coefficient (ADC) values of hepatocellular carcinoma (HCC) and pathological grade. MATERIALS AND METHODS: Preoperative magnetic resonance imaging (MRI) images of 241 patients with HCC confirmed by pathology were retrospectively analyzed. All patients underwent preoperative diffusion-weighted imaging (DWI) on a 1.5T MRI scanner. The mean and minimum ADC values of the tumors were measured. The ADC values were compared in tumors with different grades and the correlations between ADC values and pathological grade were analyzed. Receiver operating characteristic (ROC) curves of ADC values were obtained and compared to distinguish poorly and nonpoorly differentiated HCCs. Interobserver agreements were assessed by intraclass correlation coefficient (ICC). RESULTS: The mean and minimum ADC values of poorly differentiated HCCs were lower than those of nonpoorly differentiated HCCs (P = 0.000, 0.000, respectively). The mean and minimum ADC values were negatively correlated with pathological grade (rs = -0.180 and -0.202, respectively) (P = 0.005, 0.002, respectively). For the differentiation between poorly and nonpoorly differentiated HCCs, the mean ADC value provided a sensitivity of 69.57% and a specificity of 73.39% with a cutoff value of 0.96 × 10-3 mm2 /s while the minimum ADC value showed a sensitivity of 78.26% and a specificity of 61.47% with a cutoff value of 0.90 × 10-3 mm2 /s. No significant difference existed between both ROC curves (P = 0.64). The ICC for the measurements of the mean and minimum ADC values was 0.92 (95% confidence interval [CI] 0.90-0.93) and 0.91 (95% CI 0.89-0.93), respectively. CONCLUSION: DWI of HCC could preoperatively provide quantitative parameters for predicting tumor histological grade. J. Magn. Reson. Imaging 2016;44:1442-1447.