BackgroundEarly diagnosis of tuberculous meningitis (TBM) is still a challenge; the present study aimed to establish a method of detecting the antigen early secreted antigenic target 6 (ESAT-6) in cerebrospinal fluid (CSF) by an indirect enzyme-linked immunosorbance assay (ELISA) protocol and to study the value of detecting ESAT-6 in CSF in the early diagnosis of TBM.MethodsAn indirect ELISA protocol was used, employing a monoclonal antibody (mAb) against ESAT-6, which was used to demonstrate ESAT-6 in the CSF from TBM patients and non-TBM controls. CSF was obtained from 100 patients: confirmed TBM, clinically diagnosed TBM, disease controls, and healthy controls (n = 10). Pure recombinant ESAT-6 (standard product) was used in serial dilutions to detect the absorbance and to establish a standard curve from the data; the concentration was on the X axis vs. absorbance on the Y axis, and the standard curve was used to interpolate the concentration of ESAT-6 in samples.ResultsThe indirect ELISA method provided 88 % sensitivity and 92 % specificity for the diagnosis of TBM using a mAb to ESAT-6. The mean concentration of ESAT-6 in TBM patients was significantly higher than that of the non-TBM groups. There was also a significant difference in the mean ESAT-6 expression between the confirmed TBM patients and the clinically diagnosed TBM patients (p < 0.01).ConclusionsDetection of ESAT-6 in the CSF of TBM patients by indirect ELISA protocol gives a reliable early diagnosis and can be used to develop an immunodiagnostic assay with increased sensitivity and specificity.

The aim of the study was to investigate the effect of astaxanthin and its possible mechanisms on acute cerebral infarction (ACI) in rat model. Male Sprague Dawley rats were randomly divided into sham group, model group, and astaxanthin-treated groups (20, 40, and 80 mg/kg). Neurological examination, the ratio of cerebral edema, and histopathology changes were assessed. Moreover, some oxidative stress markers were obtained for biochemical analysis, and the expression of neurotrophic factors gene was detected by real-time polymerase chain reaction (RT-PCR) method. The results showed that treatment with astaxanthin notably reduced neurological deficit scores and the ratio of cerebral edema compared with the model group. Meanwhile, astaxanthin increased the activity of catalase, superoxide dismutase, and glutathioneperoxidase as well as decreased the content of malondialdehyde in brain tissue. RT-PCR results showed that the expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) mRNA were increased with astaxanthin treatment. The results indicated that astaxanthin could ameliorate ACI followed by suppressing oxidative stress and upregulating the expression of BDNF and NGF mRNA.

Introduction: The polarization state of microglia affects the progress of neuropathic pain. MiR-155 modulates polarization of microglia, while its role in neuropathic pain has not been well studied. Material and methods: We separately used lipopolysaccharide (LPS) and interleukin 4 (IL-4) for constructing an M1/M2 polarization model in BV-2 cells. The levels of CD86, iNOS, CD206, Arg and miR-155 were measured by western blot or qRT-PCR, as needed. Subsequently, BV-2 cells were transfected with miR-155 mimics or inhibitor to explore the effects of miR-155 on polarization states. We also constructed a neuropathic pain model by applying spinal nerve ligation (SNL) in Wistar rats with miR-155 agomir or antagomir injection. The withdrawal threshold was measured by Von Frey fibre needle. The levels of interleukin 1β (IL-1β), tumour necrosis factor α (TNF-α) and the proportion of M1-polarized microglia in primary microglia from rats were measured by ELISA and flow cytometry. Results: LPS induced M1 polarization in BV-2 cells with increasing of CD86, iNOS and miR-155, while IL-4 induced M2 polarization in BV-2 cells with increasing of CD206, Arg and decreasing of miR-155. MiR-155 mimics upregulated CD86 and downregulated CD206, whereas miR-155 inhibitor downregulated CD86 and upregulated CD206. MiR-155 antagomir increased the withdrawal threshold, decreased the production of IL-1β, TNF-α and the proportion of M1-polarized microglia in primary microglia. Conclusions: Results demonstrate that suppression of miR-155 attenuates neuropathic pain by inducing an M1 to M2 switch in microglia. Our findings provide a new perspective to understand the function of miR-155 in microglia.

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