Focus on autonomic dysfunctions in anti-NMDAR encephalitis: a case report.

OBJECTIVE: We hope it will provide a reference for early detection, early diagnosis, and early treatment of atypical Anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis with non-typical autonomic dysfunctions as the first symptom. PATIENTS AND METHODS: We present a 15-year-old girl with the repetition of conscious disturbance at different levels, but no abnormal movements. Initially, there were no positive findings on routine electroencephalography (EEG) and dynamic video-electroencephalography (V-EEG), but the head-up tilt test (HTT) suggested neurocardiogenic syncope (vascular rejection type), which seemed to be the final diagnosis. However, the patient later experienced several episodes of disturbance of consciousness with unexplained abdominal pain. Abnormalities were discovered on EEG, which indicated the possibility of "epileptic seizures with autonomic-gastrointestinal features". Based on these findings, we finally tested the autoimmune encephalitis-related antibodies for the patient after the literature search and review. RESULTS: The patient was finally diagnosed with anti-NMDAR encephalitis. Her symptoms were fully controlled after glucocorticoid and gamma globulin treatment, and she left the hospital with complete recovery. CONCLUSIONS: Although autonomic nervous dysfunction occurred in our patient, her prognosis was good because she did not have respiratory or (and) circulatory failure. Exclusive diagnosis and early treatment are important in patients with anti-NMDAR encephalitis. Abdominal pain with positive HTT may be a manifestation of autonomic dysfunction in this disease.

Retigabine protects the blood-brain barrier by regulating tight junctions between cerebral vascular endothelial cells in cerebral ischemia-reperfusion rats.

OBJECTIVE: To investigate the effect of retigabine on the blood-brain barrier permeability in rats with cerebral ischemia-reperfusion and its mechanism. MATERIALS AND METHODS: A total of 90 Sprague-Dawley (SD) rats were selected to prepare a rat model of focal cerebral ischemia-reperfusion. The blood flow changes were detected using a laser Doppler flow meter, the percentage of the cerebral infarction volume was measured by means of the triphenyl tetrazolium chloride (TTC) staining, the effect of retigabine on the permeability of the blood-brain barrier after cerebral ischemia-reperfusion was examined via Evans blue (EB) staining, and the state of tight junctions between endothelial cells was determined via the transmission electron microscopy (TEM) technique. Immunohistochemistry was used to detect the effects of retigabine on the distribution and expressions of tight junction-associated proteins in the cerebral ischemia-reperfusion blood-brain barrier. Western blotting was adopted to examine the changes in the expressions of related proteins in cerebral ischemia-reperfusion tissues. RESULTS: At 48 h and 96 h after cerebral ischemia-reperfusion, retigabine notably reduced the cerebral infarction volume of rats, and the tight junctions between microvascular endothelial cells in the ischemic area opened up, the permeability of the blood-brain barrier was remarkably increased, and the permeability of the blood-brain barrier was significantly reduced under the action of retigabine. The expressions of claudin-5, occludin, and ZO-1 in the blood-brain barrier of the ischemic brain tissue significantly declined, and retigabine notably increased the expressions of three proteins and their distributions along the microvessels. At 3 h, 24 h, 48 h, and 96 h after cerebral ischemia-reperfusion, the expressions of the MMP-2 protein and MMP-9 protein in the ischemic brain tissue were evidently increased, which were inhibited by retigabine. Moreover, the expressions of the PKCδ protein in the ischemic brain tissue were markedly increased, which were significantly inhibited by retigabine. CONCLUSIONS: The regulatory roles of retigabine in the distribution and expressions of claudin-5, occludin, and ZO-1 may be associated with the inhibition of the expressions of the MMP-2, MMP-9, and PKCδ proteins.

Dl-3-n-butylphthalide protects the blood brain barrier of cerebral infarction by activating the Nrf-2/HO-1 signaling pathway in mice.

OBJECTIVE: The aim of this study was to explore whether Dl-3-n-butylphthalide (DBT) could protect blood-brain barrier (BBB) of mice with experimental cerebral infarction and the relevant mechanism. MATERIALS AND METHODS: Adult male CD-1 mice were selected as the study objects. The permanent middle cerebral artery occlusion (MCAO) model was prepared by Longa's modified suture-occluded method. The mice were randomly divided into 3 groups: the sham operation group (Sham group), the cerebral infarction model group (CI group) and the DBT (120 mg/kg) intervention group (DBT group). Neurologic function deficits were evaluated by Longa's modified scoring method after 24 h of permanent MCAO. The wet and dry weight method was used for measuring water content in brain tissues. 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining method was applied to determine the volume of cerebral infarction. Changes in the protein and messenger ribonucleic acid (mRNA) expression levels of matrix metallopeptidase 9 (MMP-9), claudin-5, vascular endothelial growth factor (VEGF), glial fibrillary acidic protein (GFAP), NF-E2 related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1) in ischemic brain tissues were detected using immunohistochemistry, Western blotting and quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Ultrastructure changes in BBBs were observed under an electron microscope. RESULTS: DBT improved the neurologic function deficits of mice and reduced the infarction volume of mice with cerebral infarction. DBT alleviated edema and decreased the permeability of BBBs of mice with cerebral infarction. DBT down-regulated the expression of MMP-9 and up-regulated the expression of claudin-5 in brain tissues of mice with cerebral infarction. DBT increased the expressions of VEGF and GFAP. DBT improved the ultrastructure in capillary endothelial cells of BBBs and increased the expressions of Nrf-2 and HO-1. CONCLUSIONS: DBT may protect BBB by activating the Nrf-2/HO-1 signaling pathway, thus achieving its protective effect on the brain.

Relationship between expression level of hygromycin B-resistant gene and Agrobacterium tumefaciens-mediated transformation efficiency in Beauveria bassiana JEF-007.

AIMS: Agrobacterium tumefaciens-mediated transformation (AtMT) is an effective method for generation of entomopathogenic Beauveria bassiana transformants. However, some strains grow on the selective medium containing hygromycin B (HygB), which reduces the selection efficiency of the putative transformants. In this work, a relationship between HygB resistance gene promoter and AtMT efficiency was investigated to improve the transformant selection. METHODS AND RESULTS: Ten B. bassiana isolates were grown on 800 µg ml-1 HygB medium, but only JEF-006, -007 and -013 showed susceptibility to the antibiotics. Particularly, JEF-007 showed the most dose-dependent susceptibility. Two different Ti-Plasmids, pCeg (gpdA promoter based) and pCambia-egfp (CaMV 35S promoter based), were constructed to evaluate the promoters on the expression of HygB resistance gene (hph) at 100, 150 and 200 µg ml-1 HygB medium. Eight days after the transformation, wild type, AtMT/pCeg and AtMT/pCambia-egfp colonies were observed on 100 µg ml-1 HygB, but significantly larger numbers of colonies were counted on AtMT/pCeg plates. At higher HygB concentration (150 µg ml-1 ), only AtMT/pCeg colonies were further observed, but very few colonies were observed on the wild type and AtMT/pCambia-egfp plates. Putative transformants were subjected to PCR, RT-PCR and qRT-PCR to investigate the T-DNA insertion rate and gene expression level. Consequently, >80% of colonies showed successful AtMT transformation, and the hph expression level in AtMT/pCeg colonies was higher than that of AtMT/pCambia-egfp colonies. CONCLUSIONS: In the HygB-susceptible B. bassianaJEF-007, gpdA promoter works better than CaMV 35S promoter in the expression of HygB resistance gene at 150 µg ml-1 HygB, consequently improving the selection efficiency of putative transformants. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide useful information for determining AtMT effectiveness in B. bassiana isolates, particularly antibiotic susceptibility and the role of promoters.

Performance evaluation of the small-animal PET scanner ClairvivoPET using NEMA NU 4-2008 Standards.

The aim of this study was to evaluate the performance of ClairvivoPET using NEMA NU4 standards. The ClairvivoPET incorporates a LYSO dual depth-of-interaction detector system with 151 mm axial field of view (FOV). Spatial resolution, sensitivity, counting rate capabilities, and image quality were evaluated using NEMA NU4-2008 standards. Normal mouse imaging was also performed for 10 min after intravenous injection of (18)F(-)-NaF. Data were compared with 19 other preclinical PET scanners. Spatial resolution measured using full width at half maximum on FBP-ramp reconstructed images was 2.16 mm at radial offset 5 mm of the axial centre FOV. The maximum absolute sensitivity for a point source at the FOV centre was 8.72%. Peak noise equivalent counting rate (NECR) was 415 kcps at 14.6 MBq ml(-1). The uniformity with the image-quality phantom was 4.62%. Spillover ratios in the images of air and water filled chambers were 0.19 and 0.06, respectively. Our results were comparable with the 19 other preclinical PET scanners based on NEMA NU4 standards, with excellent sensitivity because of the large FOV. The ClairvivoPET with iterative reconstruction algorithm also provided sufficient visualization of the mouse spine. The high sensitivity and resolution of the ClairvivoPET scanner provided high quality images for preclinical studies.

Early diagnosis of tuberculous meningitis by an indirect ELISA protocol based on the detection of the antigen ESAT-6 in cerebrospinal fluid.

BACKGROUND: Early 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. METHODS: An 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. RESULTS: The 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). CONCLUSIONS: Detection 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.

Impact of operating conditions on nitrogen removal using cyclic activated sludge technology (CAST).

In this study, the nitrogen removal performance of a municipal wastewater treatment plant that employs cyclic activated sludge technology (CAST) was evaluated. The impact of key operational conditions such as temperature, dissolved oxygen (DO) concentrations and operating modes were examined in parallel. During summer, when the operating temperature ranged from 27-30 degrees C, the NH(4)(+)-N and total nitrogen (TN) removal efficiencies were 51 +/- 7% and 42 +/- 7%, respectively, and simultaneous nitrification and denitrification (SND) was considered to be the major nitrogen removal mechanism. In contrast, at a low operating temperature of 10-15 degrees C, the DO concentration increased from 0.7 to 3.0 mg/L; however, the NH(4)(+)-N and TN removals were both inefficient due to poor biomass activities, which was demonstrated by a lower specific oxygen uptake rate (SOUR) of 1.2 mg O(2)/g SS. h (15 degrees C). Moreover, a 3h-mode for the CAST process was preferable during winter because the effluent wastewater quality was similar to that obtained when the 4h-mode was used. The extremely low organic loading was the primary reason for the poor bioactivity of the sludge in the CAST system, and this eventually led to deterioration of the nitrogen removal efficiencies.