The radial migration of cortical pyramidal neurons (PNs) during corticogenesis is necessary for establishing a multilayered cerebral cortex. Neuronal migration defects are considered a critical etiology of neurodevelopmental disorders, including autism spectrum disorders (ASDs), schizophrenia, epilepsy, and intellectual disability (ID). TRIO is a high-risk candidate gene for ASDs and ID. However, its role in embryonic radial migration and the etiology of ASDs and ID are not fully understood. In this study, we found that the in vivo conditional knockout or in utero knockout of Trio in excitatory precursors in the neocortex caused aberrant polarity and halted the migration of late-born PNs. Further investigation of the underlying mechanism revealed that the interaction of the Trio N-terminal SH3 domain with Myosin X mediated the adherence of migrating neurons to radial glial fibers through regulating the membrane location of neuronal cadherin (N-cadherin). Also, independent or synergistic overexpression of RAC1 and RHOA showed different phenotypic recoveries of the abnormal neuronal migration by affecting the morphological transition and/or the glial fiber-dependent locomotion. Taken together, our findings clarify a novel mechanism of Trio in regulating N-cadherin cell surface expression via the interaction of Myosin X with its N-terminal SH3 domain. These results suggest the vital roles of the guanine nucleotide exchange factor 1 (GEF1) and GEF2 domains in regulating radial migration by activating their Rho GTPase effectors in both distinct and cooperative manners, which might be associated with the abnormal phenotypes in neurodevelopmental disorders.
Autism Spectrum Disorder/metabolism* ; Cell Movement/genetics* ; Humans ; Interneurons/metabolism* ; Neurodevelopmental Disorders/genetics* ; Neurons/metabolism* ; Rho Guanine Nucleotide Exchange Factors/genetics*

Objective To establish a test of autoantibody-panel for the diagnosis of autoimmune cerebellitis (AC) and determine the prevalence of AC in patients with cerebellar ataxia of unknown etiology.Methods Autoantibody screening tests with indirect immunofluorescence were performed in serum and cerebrospinal fluid (CSF) samples of 400 previously'idiopathic'Chinese patients with cerebral ataxia (inpatients and outpatients in Peking Union Medical College Hospital or referred from hospitals of Beijing Encephalitis Group from 2016 to 2018).Immunotherapy was given to autoantibody positive patients and the effectiveness of immunotherapy was assessed.Detailed AC autoantibodies panel included anti-glutamate decarboxylase 65 (GAD65) antibody,anti-Tr (delta notch-like epidermal growth factor-related receptor (DNER)) antibody,anti-zinc finger protein 4 (ZIC4) antibody,anti-inositol 1,4,5-trisphosphate receptor 1 (ITPR1) antibody,anti-homer protein homolog 3 (Homer 3) antibody,anti-neurochondrin (NCDN) antibody,anti-carbonic anhydrase-related protein (CARP) antibody and anti-Purkinje cell antibody 2 (PCA2) antibody.Results Eight out of 400 (2％) ataxia patients were positive for this AC panel tests,of whom two were positive for anti-GAD65 antibody,two for anti-Tr antibody,one for anti-PCA2 antibody,one for anti-Homer 3 antibody and two were positive for serum anti-NCDN antibody.Autoantibodies against ZIC4,ITPR1 and CARP were not detected in this cohort.Two of the eight ataxia patients also presented with limbic encephalitis,and only one anti-GAD antibody patient was screened with underlying small cell lung carcinoma (SCLC).All the eight patients received immunotherapy and four experienced partial response.Conclusions Autoimmune cerebellitis is the cause of acquired cerebellar ataxia.Tests of autoantibodies associated with AC have diagnostic value for paraneoplastic and non-paraneoplastic cerebellar ataxia.Immunotherapy may yield partial response in patients with AC.

Objective To explore the management of spontaneous intraspinal hematoma.Methods From January 2011 to July 2018,29 cases with spontaneous intraspinal hematoma were admitted to our department.Date on etiology,clinical presentation,radiological features,treatment strategy and prognosis were analyzed retrospectively.The prognosis was assessed by American Spinal Injury Association impairment scale (ASIA) before and after the treatment.Results Total of 29 cases,only 10 cases (34.5％) revealed specific etiology,including 7 cases of spinal vascular malformation,2 of tumor apoplexy,1 of cavernous hemangioma.After 2 weeks of conservative treatment,3 patients with grade D and 3 patients with grade E were assessed for spinal function.The average interval from onset to surgery was(9.4±7.5) days,the ASIA after two weeks of the operation was as follows:5 patients were assessed at grade A,5 patients at grade C,8 patients at grade D and 4 patients at grade E.28 patients were followed up for (48.7±23.1) months on average,6 patients without surgery were E,22 cases with surgery were as follows:4 cases A,18 cases D/E.Conclusions The etiology of spontaneous intraspinal hematoma is hard to define even after complete preoperative examination and exploratory operation.The preoperative neurologic functions are important predicting factors for the prognosis of spontaneous intraspinal hematoma.For patients who had neurologic function deficit,surgical treatment should be performed urgently to remove the hematoma and release the decompression of spinal cord.The majority of these patients can achieve a positive prognosis after surgery.

Objective To assess effects of stress dynamic CT myocardial perfusion imaging (CT-MPI) combined with coronary CT angiography (CCTA) on the diagnosis of myocardial perfusion defects in coronary artery disease (CAD). Methods Patients with CAD diagnosed by CCTA underwent ATP stress CT-MPI examination. Single-photon emission computed tomography (SPECT) myocardial perfusion imaging (SPECT-MPI) was performed within one week and set as the reference standard. CT-MPI results were qualitatively analyzed, and myocardial blood flow (MBF), myocardial blood volume (MBV) as well as time to peak (TTP) were quantified according to CT-MPI. Effects of CCTA, CT-MPI, and CT-MPI combined with CCTA on predicting myocardial perfusion defects were assessed in comparison with NMPI. Results Thirty patients [(54.8±8.4)years] were enrolled in our study, 20 were men (68%). MBF [(79.3±18.0) versus (135.1± 35.2) ml·100 ml-1·min-1] and MBV [(8.9±2.9) versus (13.8±8.9) ml/100 ml] were significantly decreased in hypoperfused segments compared with normal segments, while TTP was increased in hypoperfused segments [(13.9 ± 2.5)s] compared with normal segments [(9.1 ± 2.1)s] (t=0.302, 0.866 and 0.024 respectively, all P values<0.01). The sensitivity, specificity of CT-MPI for identifying segments with perfusion defects were 91.3%(147/161), 84.6%(281/332), respectively. On a per-vessel basis, the area under the receiver operating characteristic curve for predicting myocardial perfusion defects were 0.635(95%CI:0.517—0.753) for CCTA, 0.709(95%CI:0.599—0.819)for CT-MPI, and 0.837(95%CI:0.749—0.925)for CT-MPI combined with CCTA, respectively. Conclusions The performance of stress dynamic CT-MPI in the diagnosis of myocardial perfusion defects in CAD was good. One-stop examination of CT-MPI combined with CCTA improves the diagnostic accuracy for identifying flow-obstructing stenosis.

A method for quantitative detection of florfenicol by colloidal gold lateral flow immunoassay was developed.The experimental conditions including pH value, concentrations of antibody in the process of conjugation between the colloidal gold and antibody, amount of gold-labeled antibody, concentration of the antigen sprayed on test lines (T line), and detection time were optimized.With a colloidal gold strip reader, the signal intensity of T lines and control lines (C line) on lateral flow strips was recorded.The T/C ratio of negative control and positive samples was defined as B0 and Bx, and the standard curve was established by plotting the Bx/B0 ratio against the concentration of florfenicol.This assay showed a good linear range from 0.1 to 1.5 ng/mL with the limit of detection of 0.08 ng/mL, while the result could be obtained within 15 min.The result showed that this quantitative method was convenient and rapid, and could be used in screening a large amount of samples on site.

Objective:To establish the quality standard for Tangzhixiao capsules. Methods: The qualitative identification of Salvia miltiorrhiza,Atractylodes and Hawthorn was detected by TLC, and the quantitative determination of salvianolic acid B was determined by HPLC. The HPLC determination was performed on a YMC-Triart C18 (250 mm × 4. 6 mm, 5μm ) column with the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (24∶ 76) at the flow rate of 1. 0 ml·min-1 , the column temperature was 30℃ and the detection wavelength was set at 286 nm. Results: The TLC spots were fairly clear, and the negative samples showed no interference. The concentration of salvianolic acid B within the range of 0. 012-0. 120 mg·ml -1 was linear with peak area(r = 0. 999 5), the average recovery was 100. 06% and RSD was 0. 83%(n = 6). Conclusion: The method is accurate, reliable and reproducible, which can be used in the quality control of Tangzhixiao capsules.

Objective To study the role of TLR2 and TLR4 signal transduction in RAW264.7 monocyte-macrophages stimulated by Aspergillus fumigatus conidia,and to investigate the expression of TLR2 signal transduction after silencing gene of TLR4.Methods Macrophages were randomly divided into normal group ( N group),normal+stimulated with Aspergillus fumigatus conidia ( N +Af group ),normal + transfected with TLR4-siRNA [ TLR4 (RNAi) group ],normal+transfected with TLR4-siRNA +stimulated with Aspergillus fumigatus conidia[ TLR4(RNAi) +Af group].RT-PCR and Western blot were used to assay expression levels of TLR2,TLR4,MyD88 mRNA and pro-inflammatory cytokines TNF-α protein when macrophages were stimulated 12 h by Aspergillus fumigatus conidia after tranfected 24 h with TLR4-siRNA by technology of RNAi.Results ( 1 ) Compared with N group,the expression of TLR2,TLR4,MyD88 mRNA and TNF-αprotein in N+Af group significantly increased before silencing gene of TLR4.(2) Silencing efficiency of macrophates was up to 83％ after transfected with TLR4-siRNA.(3)The expression of TLR2,MyD88 mRNA in TLR4 (RNAi) group significantly decreased contrast with normal group.Meanwhile the expression of TLR2,MyD88 mRNA and TNF-α protein also obviously reduced in TLR4(RNAi) +Af group when compared with N +Af group.Compared with TLR4 (RNAi) group,the expression of MyD88 mRNA in TLR4 (RNAi) +Af group significantly increased.However,the expression of TLR2 mRNA and TNF-α protein have no significant change after silencing gene of TLR4.Conclusion Signaling pathway of TLR2 and TLR4 in macrophages was activated by given stimulus of Aspergillusfumigatus conidia and exerted the effect of anti-Aspergillus fumigatus spores stimulation through the release of pro-inflammatory cytokines TNF-α.Meanwhile,silencing gene of TLR4 down-regulate the effect of TLR2 signal transduction in RAW264.7 cells to anti-Aspergillus fumigatus conidia stimulation,and it found that TLR4 played an more important role by contrast with TLR2.

Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.

OBJECTIVETo prepare berberine microemulsion, and to investigate its properities and the absorption character in rat intestine in situ.

METHODThe optimum formulation of the blank microemulsion selected by pseudo tertiary phase diagrams and the berberine microemulsion was prepared based on the blank microemulsion. The viscosity, conductance, refraction rate and particle size of berberine microemulsion were surveyed. An in situ rat perfusion method was used to investigate the intestinal absorption of berberine microemulsion. A UV method for determination of berberine in the intestinal flux was established.

RESULTThe viscosity, conductance, refraction rate and particle size of berberine microemulsion were 2.11 cPas, 125.5 microomega, 1.363 and 24.0 nm, respectively. The absorption rate of berberine at the ileum was the best. The absorption of berberine microemulsion at the ileum was significantly higher than that of raw medicine (P < 0.01).

CONCLUSIONThe microemulsion system might improve the absorption of berberine in the intestinal tract.

Absorption ; Administration, Cutaneous ; Animals ; Berberine ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; Drug Stability ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Intestines ; metabolism ; Male ; Particle Size ; Rats ; Rats, Wistar ; Skin ; metabolism ; Skin Absorption ; drug effects ; physiology ; Solubility ; Technology, Pharmaceutical ; methods

can specifically combined with EGFR, which may be applied to noninvasive NIRF imaging of tumors highly expressed EGFR in vivo.