[Diagnostic performance of a novel Mycobacterium Tuberculosis specific T-Cell based assay for tuberculosis].

Objective: To evaluate the diagnosic performance of a novel Mycobacterium tuberculosis (MTB) specific T-cell based assay for tuberculosis, which targets the mRNA detection of interferon gamma-induced protein 10 (IP-10). Methods: Suspected tuberculosis patients were prospectively and consecutively recruited in Beijing Chest Hospital between March 2018 and November 2019, and individuals with lower risk of MTB infection were also recruited. IP-10.TB and T-SPOT.TB assays were simulataneously performed on peripheral blood samples. The diagnostic performance of IP-10.TB and T-SPOT.TB were analyzed using the receiver operating characteristic curve. Accordance of IP-10.TB and T-SPOT.TB was analyzed by Cohen's kappa test, while the correlation between the expression level of IP-10 mRNA in IP-10.TB test and the number of SFCs in T-SPOT.TB test were analyzed by Pearson correlation test. Results: A total of 235 patients with tuberculosis, 110 patients with other diseases and 153 individuals with lower risk of MTB infection were included in the final analysis. No significant difference was detected in the rate of indeterminate results between IP-10.TB assay (3/498, 0.60%) and T-SPOT.TB assay (6/498, 1.21%). The total sensitivity and specificity of IP-10.TB assay were 91.3% (95%CI 86.8%-94.6%) and 81.1% (95%CI 75.8%-85.7%). The specificity of IP-10.TB in individuals with lower risk of MTB infection was 98.0% (95%CI 94.4%-99.6%). The total sensitivity and specificity of T-SPOT.TB assay were 93.0% (95%CI 88.9%-96.0%) and 83.8% (95%CI 78.7%-88.1%). The specificity of T-SPOT.TB in individuals with lower risk of MTB infection was 100% (95%CI 97.6%-100.0%). No significant differences were detected in sensitivity and specificity between IP-10.TB and T-SPOT.TB assays (P>0.05). The positive coincidence rate of these 2 methods was 91.0% (95%CI 87.5%-94.5%), and the negative coincidence rate was 88.9% (95%CI 84.9%-92.9%) and the total coincidence rate was 90.0% (95%CI 87.3%-92.6%). The Cohen's kappa value was 0.80 (95%CI 0.75-0.85, P<0.001) between IP-10.TB and T-SPOT.TB assays. Conclusion: These results showed that the diagnostic performance of IP-10.TB was consistent with that in T-SPOT.TB, and this test could be a novel adjunctive tool for the diagnosis of tuberculosis.

Molecular regulation of organic anion transporting polypeptide 1A2 (OATP1A2)by taurocholic acid in Bewo Cells.

To characterize the mechanisms of action of taurocholic acid(TCA) and farnesoid X receptor(FXR) on organic anion transporting polypeptide 1A2(OATP1A2) expression in placental Bewo cell line. Quantitative real-time PCR and Western blots were used to detect OATP1A2 in Bewo cells cultured with TCA and pcDNA3.1(+)-hFXR transfected Bewo cells after incubation with 2 mM TCA for 48 hours. TCA(0.02 mM) induced the mRNA and protein expression of OATP1A2 by 3 and 1.6 fold (p<0.05), respectively, while 0.2 and 2 mM TCA induced mRNA and protein expression by 1.5 and 1.3 fold, respectively. The concentration of TCA was negatively correlated with OATP1A2 gene expression (P<0.05). In pcDNA3.1(+)-hFXR transfected Bewo cells with 2 mM TCA demonstrated a 2-3 fold increase in OATP1A2 over controls (P<0.05). TCA is one of the regulation factors for OATP1A2 in the Bewo cell line. A low dose of TCA can induce fetal membrane expression of OATP1A2. This may present a physiological or compensatory mechanism of the placenta, while the high dose of TCA may produce a pathological or pathogenic mechanism. Farnesoid X receptor may act in synergy with TCA to increase the expression of OATP1A2. This may be a treatment strategy for fetal cholestasis.

Oxidative stress markers in intrahepatic cholestasis of pregnancy: a prospective controlled study.

OBJECTIVE: Intrahepatic cholestasis of pregnancy (ICP), characterized by skin pruritus and elevation of serum aminotransferase activity and bile acid concentration in the mother, is one of the most common liver disorders in pregnancy. It was proved that ICP might lead to fetal distress by triggering oxidative damage. Total bile acids (TBA) are an established marker for assessment of the severity of ICP. The aim of this study was to explore associations of TBA levels with levels of the oxidative stress markers 8-epimer of prostaglandin F2alpha (8-iso-PGF2α), superoxide dismutase (SOD) and glutathione peroxidase (Gpx) in ICP. PATIENTS AND METHODS: Maternal plasma levels of 8-iso-PGF2α, SOD and Gpx were examined in ICP patients (n=40) and normal pregnancy controls (n=47) using an enzyme-linked immunosorbent assay (ELISA) analysis. RESULTS: Plasma levels of 8-iso-PGF2α and Gpx were significantly lower in ICP patients than in controls (p = 0.006 and 0.002, respectively), while no significant difference was observed in SOD levels between the two groups. Levels of 8-iso-PGF2α and TBA were negatively correlated (r = -0.277, p = 0.01, Spearman's correlation coefficient). CONCLUSIONS: The clinical severity of ICP is closely related to the degree of lipid peroxidation, and the natural antioxidant system might fail to work effectively in the presence of lipid peroxidation damage in ICP.

Unexpected expression of glucose transporter 4 in villous stromal cells of human placenta.

Glucose transporter 4 (GLUT4) protein expression was characterized in human and rodent term placentas. A 50-kDa protein was detected, by immunoblotting, in term human placenta at levels averaging 25% of those found in white adipose tissue. It was also present, albeit at lower levels, in mouse and rat placentas. The specificity of the 50-kDa signal was established by using skeletal muscle and placental tissues obtained from GLUT4-null mice as controls. Indirect immunohistochemistry, performed in human placentas, showed that intravillous stromal cells were conspicuously labeled by GLUT4 and revealed colocalization of GLUT4 transporters with insulin receptors. This study provides the first evidence that the insulin-responsive GLUT4 glucose transporter is present in human and rodent hemochorial placentas. Placental GLUT4 gene and protein levels were not modified in human pregnancy complicated by insulin-dependent diabetes mellitus. The significance of the high level of GLUT4 protein in human placenta remains to be elucidated, because, so far, this organ was not considered to be insulin-sensitive, with regard to glucose transport.

Inadequate values from an interferon-gamma release assay for smear-negative tuberculosis in a high-burden setting.

OBJECTIVE: To examine the usefulness of an interferon-gamma release assay (IGRA) for the diagnosis of smear-negative tuberculosis (TB) in China. DESIGN: A total of 624 patients with presumed pulmonary TB were enrolled prospectively and categorised as smear-negative TB, smear-positive TB or no TB. All patients were tested using T-SPOT.TB. RESULTS: Both the smear-negative and smear-positive TB groups had significantly more spot-forming cells (SFCs) than the no TB group (all P < 0.001), while the smear-negative group had fewer SFCs than the smear-positive TB group (P < 0.001). The specificity of T-SPOT.TB was 60.4% (95%CI 53.4-67.1). The sensitivities of T-SPOT.TB in the smear-negative and smear-positive TB groups were respectively 81.4% (95%CI 75.7-86.0) and 93.2% (95%CI 87.6-96.4). The sensitivity in the smear-negative TB group was much lower than that in the smear-positive TB (P < 0.05). CONCLUSIONS: The sensitivity of T-SPOT.TB was lower due the paucibacillary nature of the samples, and the specificity was lower due to the high prevalence of latent tuberculous infection in the smear-negative TB patients. The T-SPOT.TB test should only be used as a supplementary test and not as a single test to rule in or rule out smear-negative TB.

The molecular mechanism of microRNA-145 to suppress invasion-metastasis cascade in gastric cancer.

Invasion and metastasis are the major features of malignant tumors that are responsible for 90% of cancer-related deaths. Recently, microRNAs have been discovered to have a role in suppressing tumor metastasis. This study's aim was to clarify the roles of miR-145 in gastric carcinomas and its underlying molecular mechanism in regulating tumor metastasis. Here, we demonstrate a stepwise downregulation of miR-145 level in nontumorous gastric mucosa, primary gastric cancers and their secondary metastases. In vitro analysis of miR-145's ectopic expression and loss-of-function suggests that it suppresses gastric cancer cell migration and invasion. In vivo spontaneous metastasis and experimental metastasis assay further confirm its function in suppressing the invasion-metastasis cascade, including impairing local invasion and inhibiting hematogenous metastasis in gastric cancers. Furthermore, we identified a novel mechanism of miR-145 to suppress metastasis. N-cadherin (CDH2) was proved to be a direct target of miR-145, using luciferase assay and western blot. Re-expressing N-cadherin in miR-145-transfected cells reverses their migration and invasion defects. Although not a direct target of miR-145, matrix metallopeptidase 9 (MMP9), but not MMP2, was also significantly decreased in miR-145-expressing cells. We suggest that miR-145 suppresses tumor metastasis by inhibiting N-cadherin protein translation, and then indirectly downregulates its downstream effector MMP9.