Objective To establish the chromatographic fingerprint of mushroom polysaccharides by 1-phenyl-3-methyl-5-pyrazolone( PMP) pre-column derivatization. Methods The mushroom polysaccharides was extracted by hot distilled water, precipitated by alcohol, and hydrolyzed into monosaccharides by Trifluoroacetic Acid (TFA). The hydrolysate was derivatized with 1-phenyl-3-methyl-5-pyrazolone ( PMP ) and tested via HPLC to study the monosaccharide components in mushroom polysaccharides. Results Fingerprint was established with 13 common peaks, 6 peaks in which were identified as mannose, D-glucuronic acid, D-glucose, galactose, xylose and L-fucose. The glucose accounted for the most, followed by mannose, galactose and fucose. Conclusion Development of fingerprint chromatogram by HPLC is a stable, simple, and repeatable way, which can be applied to the quality control of mushroom polysaccharides.

AIM:To investigate effect of hypoxia on the expression of proliferating cell nuclear antigen(PCNA) a nd phenotype of cardiac fibroblasts(CFs). METHODS: The purif ied cardiac fibro blasts were cultured and divided randomly into there groups :control group, mode rate hypoxia(MH) group and severe hypoxia(SH) group. After 72 h,MTT method was u s ed to investigate the proliferation of CFs, and the ultrastructure of fibroblast s were observed with transmission electron microscopy The expression of PCNA a n d ?-actin in cardiac fibroblasts were measured by the means of immunohistochemi s try and laser scanning confocal microscopy, respectively. RESULTS: MTT A 490 nm value of MH group was significantly higher than that of control group by (18 4?25 0)% ( P

Objective Mitotic spindle and midbody are both microtubule-based temporary structures during cell growth and play essential roles in mitosis.The purpose of this study was to establish a mature and efficient method to extract mitotic spindle and midbody.Methods Through the cell cycle synchronization method,mitotic spindle or midbody was made appear inside cells.Low permeability swelling and glycerol gradient centrifugation principles were then used to extract spindle and midbody.Results By Western Blot and immunofluorescence staining,the extracts were identified as mitotic spindle and midbody.Conclusions The successful extraction of mitotic spindle and midbody from synchronized Hela cells will provide foundation for identifying the proteins located in cell during mitosis,and be of great significance to the study of molecular regulation mechanisms of mitosis and tumorigenesis.

OBJECTIVETo detect the anti-enterovirus 71 (EV71) IgM level in cerebrospinal fluid (CSF) of children with severe hand, foot and mouth disease (HFMD) induced by EV71 and then analyze the relationships among the IgM antibody levels, CSF routine examination and patients' clinical features, and thus to evaluate the clinical significance of anti-EV71 IgM as a new indicator for early diagnosis of children with severe HFMD induced by EV71.

METHODA total of 294 laboratory-confirmed cases of children with severe HFMD infected with EV71 were enrolled into the research group from March 2014 to June 2014, consisting of 53 fatal cases and 241 severe cases, and their CSF samples underwent enzyme-linked immunosorbent assay (ELISA) for anti-EV71 IgM levels, CSF routine and biochemical tests. Forty-one cases of children with severe HFMD induced by other enteroviruses were collected as antibody-testing control group during the same period.

RESULTIn the research group, the total positive rate of anti-EV71 IgM in 294 CSF samples of children with severe HFMD infected by EV71 was 60.2% (177/294); the positive rate of anti-EV71 IgM in the fatal HFMD subgroup was 62.3% (33/53); the positive rate of anti-EV71 IgM in the severe HFMD subgroup was 59.8% (144/241). In the control group, the results of CSF anti-EV71 IgM tests were all negative (0/41). In the research group, patients in antibody-positive subgroup (2.5±1.2) years old were younger than those in antibody-negative subgroup (2.9±1.1) years old (t=2.595, P=0.010). And within the antibody-positive subgroup, the patients ((1.9±0.7) years old) with fatal type disease were younger than those ((2.6±1.2) years old) with severe type disease (t=3.150, P=0.002). The CSF nucleated cells count and positive rates (105 (56,180) ×10(6) /L; 97.7% (173/177)) in antibody-positive subgroup were higher than those (62(30,150) ×10(6) /L; 83.8% (98/117)) in antibody-negative subgroup (Z=3.663, P=0.000; χ(2)=19.089, P=0.000). In antibody-positive subgroup, the percentage of monocytes (57±25)% was higher than that of polykaryocytes (43±25)%. In antibody-negative subgroup, the percentage of monocytes (50±26)% was close to that of polykaryocytes (50±26)%. In the antibody-positive subgroup, the ratio of the patients with nucleated cells count higher than 100×10(6)/L in fatal type group and severe type group was 69.7% (23/33) and 47.2% (68/144) respectively (χ(2)=5.429, P=0.02). The CSF protein quantity and positive rates in antibody-positive subgroup were higher than those in antibody-negative subgroup (Z=2.158, P=0.031; χ(2)=5.921, P=0.015).

CONCLUSIONSThe anti-EV71 IgM levels in CSF can serve as an important indicator for early diagnosis of children with severe HFMD induced by EV71. And the anti-EV71 IgM levels in CSF correlated to the CSF nucleated cells count and classification and CSF protein quantity. In the antibody-positive subgroup, the higher the nucleated cell count or the younger the age, the higher the possibility of patients to develop into fatal cases.

The human Gadd45 protein family plays critical roles in DNA repair, negative growth control, genomic stability, cell cycle checkpoints and apoptosis. Here we report the crystal structure of human Gadd45γ [corrected], revealing a unique dimer formed via a bundle of four parallel helices, involving the most conserved residues among the Gadd45 isoforms. Mutational analysis of human Gadd45γ [corrected] identified a conserved, highly acidic patch in the central region of the dimer for interaction with the proliferating cell nuclear antigen (PCNA), p21 and cdc2, suggesting that the parallel dimer is the active form for the interaction. Cellular assays indicate that: (1) dimerization of Gadd45γ [corrected] is necessary for apoptosis as well as growth inhibition, and that cell growth inhibition is caused by both cell cycle arrest and apoptosis; (2) a conserved and highly acidic patch on the dimer surface, including the important residues Glu87 and Asp89, is a putative interface for binding proteins related to the cell cycle, DNA repair and apoptosis. These results reveal the mechanism of self-association by Gadd45 proteins and the importance of this self-association for their biological function.
Amino Acid Motifs ; Animals ; Apoptosis ; radiation effects ; CDC2 Protein Kinase ; Cell Cycle ; Cell Survival ; Crystallography, X-Ray ; Cyclin B ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Cyclin-Dependent Kinases ; HeLa Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; chemistry ; genetics ; metabolism ; Mice ; Mutagenesis, Site-Directed ; Mutation, Missense ; Proliferating Cell Nuclear Antigen ; metabolism ; Protein Binding ; Protein Interaction Domains and Motifs ; Protein Multimerization ; Protein Structure, Quaternary ; Ultraviolet Rays

Objective:To discuss the feasibility of offering specialized courses of critical care medicine in undergraduate clinical medicine education, so as to alleviate the shortage of critical care medicine staffs and lay a foundation for improving the success rate for the treatment of critical cases.Methods:The undergraduates majoring in clinical medicine from 2008 to 2011 in Guizhou Medical University (the former Guiyang Medical College) were enrolled. After they had been enrolled in the undergraduate education for 3 years and were ready for Grade four, which meant basic medicine teaching had been completed and clinical medicine teaching was about to start, they were introduced and preached to each discipline, including critical care medicine. The undergraduates were free to choose professional direction of clinical training in Grade four. Students majoring in clinical medicine from 2012 to 2014 were free to choose their major direction when they entered the school.Results:From September 2011 to July 2019, the university had cultivated 246 undergraduates majoring in clinical critical care medicine from 2008 to 2014, and the critical care medicine professional team of affiliated hospital had undertaken 540 teaching hours. By July 2019, all students had graduated on time, with an employment rate of 100%. Forty students took postgraduate programs in our school and other schools, accounting for 16.3%.Conclusions:Professional education of critical care medicine in the undergraduate course of clinical medicine can mobilize students' interest in learning and subjective initiative, which is conducive to career selection. During the clinical training, the students can identify and timely cure critical care cases in the early stage, and partly alleviate the current shortage of critical care medical staffs.

Bronchiectasis is a chronic, structural lung disease characterized by chronic cough, sputum production, and decreased exercise capacity. These symptoms worsen during exacerbations and impact negatively on health-related quality of life. Airway mucus clearance and pulmonary rehabilitation play an important role in the management of bronchiectasis, despite a relatively weak evidence base. Thus, the paper reviews the physiological rationale and clinical evidence of airway clearance techniques (ACTs), application and research progress of pulmonary rehabilitation in the management of bronchiectasis.