Pharmacological activation of REV-ERBα represses LPS-induced microglial activation through the NF-κB pathway.

REV-ERBα, the NR1D1 (nuclear receptor subfamily 1, group D, member 1) gene product, is a dominant transcriptional silencer that represses the expression of genes involved in numerous physiological functions, including circadian rhythm, inflammation, and metabolism, and plays a crucial role in maintaining immune functions. Microglia-mediated neuroinflammation is tightly associated with various neurodegenerative diseases and psychiatric disorders. However, the role of REV-ERBα in neuroinflammation is largely unclear. In this study, we investigated whether and how pharmacological activation of REV-ERBα affected lipopolysaccharide (LPS)-induced neuroinflammation in mouse microglia in vitro and in vivo. In BV2 cells or primary mouse cultured microglia, application of REV-ERBα agonist GSK4112 or SR9011 dose-dependently suppressed LPS-induced microglial activation through the nuclear factor kappa B (NF-κB) pathway. In BV2 cells, pretreatment with GSK4112 inhibited LPS-induced phosphorylation of the inhibitor of NF-κB alpha (IκBα) kinase (IκK), thus restraining the phosphorylation and degradation of IκBα, and blocked the nuclear translocation of p65, a NF-κB subunit, thereby suppressing the expression and secretion of the proinflammatory cytokines, such as interleukin 6 (IL-6) and tumor necrosis factor α (TNFα). Moreover, REV-ERBα agonist-induced inhibition on neuroinflammation protected neurons from microglial activation-induced damage, which were also demonstrated in mice with their ventral midbrain microinjected with GSK4112, and then stimulated with LPS. Our results reveal that enhanced REV-ERBα activity suppresses microglial activation through the NF-κB pathway in the central nervous system.

[Clinical significance of combined detection of thinprep cytology test and high risk human papillomavirus hybrid capture 2 assay in the screening and recurrence predication of cervical intraepithelial neoplasia].

OBJECTIVE: To evaluate the clinical significance of combined detection of TCT and HR-HPV-HC2 assay in the screening and recurrence prediction of CIN. METHODS: A total of 725 patients with suspected cervical abnormalities received TCT plus HR-HPV-HC2 detection and histopathological examinations.203 CINII-III patients received TCT and HC2 detection and histopathological examinations after treated with loop electrosurgical excision procedure or cold knife conization operation. The results of pathological examinations were gold standard. The sensitivity and specificity of TCT, HR-HPV-HC2 and TCT plus HR-HPV-HC2 were analyzed. RESULTS: The missed diagnosis and misdiagnosis rate of TCT and HC2 alone were 19.23%, 12.50% and 48.15%, 74.07% and sensitivity and specificity 90.02%, 97.01% and 51.85%, 17.76% respectively. The sensitivity, missed diagnosis rate and negative predictive value were 98.25%, 1.75% and 89.70% when TCT or HR-HPV-HC2 met the CIN diagnostic criteria. The specificity and positive predictive value were 58.95% and 72.80% when TCT and HR-HPV-HC2 met the CIN diagnostic criteria. The specificity of TCT or HR-HPV-HC2 alone was 80% for the prediction of recurrence at Month 6 postoperation. The specificity of TCT plus HR-HPV-HC2 was 98.27% for the prediction of recurrence at Month 3 postoperation and the negative predictive value was 98.75% at Month 6 postoperation. The sensitivity of TCT or HR-HPV-HC2 was 97.22% for the prediction of recurrence at Month 3/6. And the negative predictive value was 99.17% at Month 6 postoperation. CONCLUSION: The combined detection of TCT and HR-HPV-HC2 may improve the sensitivity and specificity of CIN and the predication of its postoperative recurrence.

Breast cancer resistance protein expression and 5-fluorouracil resistance.

OBJECTIVE: To filtrate breast cancer resistance protein (BCRP)-mediated resistant agents and to investigate clinical relationship between BCRP expression and drug resistance. METHODS: MTT assay was performed to filtrate BCRP-mediated resistant agents with BCRP expression cell model and to detect chemosensitivity of breast cancer tissue specimens to these agents. A high performance liquid chromatography (HPLC) assay was established, and was used to measure the relative dose of intracellular retention resistant agents. RT-PCR and immunohistochemistry (IHC) were employed to investigate the BCRP expression in breast cancer tissue specimens. RESULTS: MTT assay showed that the expression of BCRP increased with the increasing resistance of 5-fluorouracil (5-Fu) (P<0.05, n=3) in the cell model, while HPLC assay indicated that the intracellular retention dose of 5-Fu was significantly correlated with the expression of BCRP (r=-0.897, P<0.05, n=3). A total of 140 breast cancer tissue specimens were collected. BCRP-positive expression was detected in forty-seven specimens by both RT-PCR and IHC. As shown by MTT assay subsequently, the resistance index (RI) of 47 BCRP-positive breast cancer tissue specimens to 5-Fu was 7-12 times as high as that of adjacent normal tissue samples. BCRP expression was related to 5-Fu resistance (R2=0.8124, P<0.01). CONCLUSION: Resistance to 5-Fu can be mediated by BCRP. Clinical chemotherapy for breast cancer patients can be optimized based on BCRP-positive expression.