The aim of this study is to develop a rapid and accurately species typing method for Brucella isolates by using High Resolution Melting (HRM ) analysis .Six pairs of primers were used according to the reference for the sequence of pur‐pose gene .Nineteen biotypes of six species Brucella standard strains were identified by PCR‐HRM analysis and this analysis was used to detect the 35 clinical isolates .Results showed Brucella amplified specific melting curves were different from con‐trasted strains with primer Bspp .The six species Brucella standard strains have own characteristic curve shape from each oth‐ers by PCR‐HRM analysis with five pairs of primers .Thirty‐five clinical isolates of Brucella have entirely consistent with PCR‐HRM curve shape with Brucella melitensis standard strains .So ,PCR‐HRM analysis methods can accurately identify Brucella strains ,especially clinical isolated Brucella melitensis ,and may be used in clinical microbiology laboratories .

GABARAP, a microtuble-associated protein, is identified to interact with GABAA receptor. Anchoring of the GABAA receptor to GABARAP helps to cluster the receptor at the synaptic termini and to mediate fast synaptic transmission. GABARAP may mediate interaction of gephyrin with the GABAA receptor to stabilize clusters by forming multimeric structures. Furthermore, GABARAP and gephyrin may play more of a role in receptor sorting and transport to the cell surface than in anchoring to the cytoplasm, because at inhibitory synapses GABARAP appears to associate with transport vesicles rather than the cell surface. The association of GABARAP with NSF (N-ethylmaleimide sensitive factor), a protein involved in intracellular vesicle transport, supports this hypothesis. We cloned cDNA encoding full-length human GABARAP by nested PCR and inserted it into eukaryon expression vector pcDNA6HA and GST fusion protein expression vector pGEX4T2. The recombinant plasmid pGEX4T2-hGABARAP was transformed into E. coli BL21, from which GST-hGABARAP fusion protein was purified after IPTG induction by affinity chromatography with glutathione Sepharose-4B column. The antiserum against GABARAP was generated by immunizing rabbits with the purified GST-hGABARAP and was purified with GST-hGABARAP coupled NHS-activated Sepherose 4 column. The purified polyclonal antibody was effective for Western blotting and immunostaining. The hGABARAP was located both in the cytoplasm and nucleus with an abundant distribution around the peripheral nucleus.
Adaptor Proteins, Signal Transducing ; biosynthesis ; genetics ; immunology ; Animals ; Antibodies ; blood ; immunology ; Antibodies, Monoclonal ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Immunization ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Objective:To evaluate the reliability and validity of the Chinese version of the Personal and Social Performance scale (PSP-CHN) in patients with schizophrenia.Methods:Totally 165 out-patients and in-patients meeting the DSM-IV-TR criteria for schizophrenia were entered in the study.Ten of subjects was included the intra-rater reliability training.The Global Assessment of Functioning Scale (GAF) was regarded as the' gold standard' to investigate the validity of PSP-CHN,and the Positive and Negative Rating Scale was used to assess the severity of disease to explore the correlative validity,in the other 155 subjects.Five to seven days after the first PSP-CHN interview,the second PSP-CHN was evaluated by another investigator to assess the test-retest reliability among 66 subjects.Twenty-seven subjects with the score of Positive and Negative Syndrome Scale (PANSS) more than 60 were followed up for 8 weeks of standardized pharmacotherapy.By the end of 8 week of treatment,the PANSS and PSP-CHN were assessed again to explore the sensitivity of PSP-CHN.Results:The internal consistency (Cronbach α=0.84) and the inter-rater reliability (κ value=0.56,ICC=0.94 for PSP-CHN total score) were good.The test-retest reliability was high [intraclass correlation coefficient (ICC) of 0.95].The scale showed good construct validity with statistically significant correlations with the Global Assessment of Functioning Scale (GAF) (ICC of 0.95).The PSP-CHN score had a good negative correlation with the PANSS total score(r=-0.79,-0.57,-0.63 and -0.71,respectively,P<0.01).After 8 week treatment,PSP-CHN total score was increased with the improvement of PANSS,and the responder showed higher increasing of PSP-CHN total score (21.2) than those partial responder(10.2),significantly.Conclusion:The Chinese version of the PSP-CHN is a convenient and valid instrument to assess the personal and social functions of stabilized and acute patients with schizophrenia.

Fifteen patients with Crohn′s disease (CD) in remission diagnosed at Shanghai Jing′an District Central Hospital from February 2018 to June 2019, and 26 matched healthy subjects were recruited. All participants underwent resting-state functional magnetic resonance imaging (fMRI) scans of hippocampus. The amplitude of low-frequency fluctuations (ALFF) was calculated to determine differences in the brain. Left hippocampus was selected as seeds for functional connectivity (FC) analysis, and the results were compared between two groups. The relationship between disease duration and ALFF/FC values in abnormal regions were analyzed with Pearson correlation. Compared with the controls, the ALFF of the left hippocampus (voxel size 32) of CD patients decreased [family-wise error correstion(FWE correction), cluster level P<0.05], and the ALFF of the left medial superior frontal gyrus (voxel size 126), left supplementary motor area (voxel size 126) and left anterior cingulate gyrus increased (voxel size 37) (FWE corrected, cluster level P<0.05). Using the left hippocampus as the seed point for the whole brain functional connectivity analysis, CD patients showed increased FC strength with the left superior temporal gyrus, left medial superior frontal gyrus, left inferior frontal gyrus (opercular part), and right supplementary motor area(FEW corrected, cluster level P<0.05). Correlation analysis did not show a significantly differences between ALFF/FC value of altered brain areas and the disease duration. It suggests that there are changes in spontaneous activities and functional connectivity in the left hippocampus in patients with CD.