Objective: To explore the structural domains of the CENP-E protein that interact with Mps1 protein.Methods: Two recombinant vectors named pEGFP-CENPE2(containing 674-1085 amino acids of CENP-E protein) and pEGFP-CENPE 3(containing 1200~2134 amino acids of CENP-E protein) were transfected into human embryo kidney 293(HEK293) cells respectively.The respective energy transfer efficiency(Ef) between either EGFP-CENPE2 and Mps1,or EGFP-CENPE3 and Mps1 were detected by FRET through selective photobleaching of the acceptors.Results: Both recombinant proteins expressed in HEK293 cells transfected by the recombinant plasmids were found to co-localize with the Mps1 protein as confirmed by confocal microscopy.The Ef between EGFP-CENPE3 and Mps1 protein was [(12.63?0.48)%,n=30] and that between EGFP-CENPE3 and Mps1 protein was [(3.17?0.21)%,n=30] as revealed by the results from FRET,the result of FRET was confirmed by co-Immunoprecipitate(CO-IP) method.When compared with that between the control and Mps1,the Ef between EGFP-CENPE3 and Mps1 was significantly higher(p

OBJECTIVETo investigate the protective effect of suppressive oligodeoxynucleotides (Sup ODN) on interferon-γ (IFN-γ) and signal transducers and activators of transcription (pSTAT4) expression of Silica-induced pulmonary inflammation in Mice.

METHODSSixty Balb/c mice were randomly divided into 4 groups, normal control group, silicious group, suppressive oligodeoxynucleotides (Sup ODN) group, control oligodeoxynucleotides (Con ODN) group. Except the normal control group injected normal saline, the rest groups were induced by the intratracheal instillation of 0.1 ml (5 g/L) of sterilized silica suspension. Sup ODN group and Con ODN group were treated by i.p. injection of 0.3 ml (1mg/mL) of suppressive or control ODN 3 h before silica administration. After 7 days, the animals were killed and levels of IFN-γ were detected by ELISA. The pathologic changes in lung tissues of mice were observed with HE staining. Expressions of IFN-γ and pSTAT4 in lung tissue were detected with immunohistochemistry and quantified by Image-Pro Plus 7.0.

RESULTSHE staining showed that the lung tissue of silicious group were damaged seriously than Sup ODN group. Compared with the normal control group (serum: (280.1±41.3) pg/ml, lung tissue: (0.249±0.373), IFN-γ increased in silicious group (serum: (886.3±81.7) pg/ml, lung tissue: (0.270±0.300) (P < 0.05). Compared with the normal control group and Con ODN group [(894.5±91.6) pg/ml], IFN-γ in the serum of Sup ODN group decreased significantly (P < 0.01). Compared with the silicious group , IFN-γ in lung tissue decreased in Sup ODN group (0.241±0.250) (P < 0.05). Compared with the normal control group (0.279±0.353), pSTAT4 in lung tissue increased significantly in silicious group (0.313±0.231) (P < 0.01). Compared with the silicious group, pSTAT4 in lung tissue decreased significantly in Sup ODN group (0.269±0.523) (P < 0.01).

CONCLUSIONSup ODN attained protective effect on Silica treated mice by suppressing expression of IFN-γ and pSTAT4.

Animals ; Female ; Inflammation ; metabolism ; Interferon-gamma ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides ; pharmacology ; Phosphorylation ; STAT4 Transcription Factor ; metabolism ; Silicon Dioxide ; toxicity

OBJECTIVETo analyze the differences of lung tissue proteins in rats exposed to silica early by using comparative proteomics method and investigate the related mechanism with the occurrence and development of silicosis.

METHODSAdult male Wistar rats were randomly divided into control group and silica-treated group. The animal model was established by intratracheal (IT) instillation with silica suspension. On the 14th day after establishment of animal model, rats were sacrificed and lung tissues were collected. The total proteins were separated by means of two-dimensional gel electrophoresis (2-DE) and the differentially expressed proteins were identified by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In addition, Western blotting was performed to verify the expression of certain candidate protein.

RESULTSEleven differential expression protein spots were tested by MALDI-TOF-MS, and six proteins were identified. The levels of cathepsin D precursor, peroxiredoxin-1 (Prx-1), heat shock cognate 71 000 protein (HSP7C), heterogeneous nuclear ribonucleoprotein A3 (hnRNPA3) and fatty acid-binding protein (epidermal, E-FABP) were up-regulated in silica-treated group with the optical density (A) values. These values were 116.50+/-12.56, 148.75+/-22.40; 40.00+/-1.63, 66.00+/-13.93; 51.25+/-7.37, 92.75+/-8.69; 83.00+/-6.48, 122.75+/-24.62; 50.75+/-6.50, 93.50+/-23.10 and 100.25+/-19.99, 142.50+/-21.21 respectively. The statistical difference was observed as compared with control group (t=-2.51, -3.71, -7.28, -3.12, -3.56 and -2.90, P<0.05). However, SEC14-like protein 3 with the A values 153.00+/-11.28, 109.75+/-18.32 was down-regulated (t=4.02, P<0.01). Western blotting showed that in the expression of Prx-1 was higher in silica-treated group (0.61+/-0.05) than that in the control (0.35+/-0.05) (t=-7.24, P<0.01) when calculating the semi-quantification of this protein using ratio of optical density.

CONCLUSION2-DE pattern of lung tissue from rats exposed to silica has been established and six differentially expressed proteins have been identified. Our study is of help for further research of the mechanisms of silicosis.

Animals ; Disease Models, Animal ; Electrophoresis, Gel, Two-Dimensional ; Environmental Exposure ; Lung ; metabolism ; pathology ; Male ; Proteomics ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Silicosis ; metabolism

OBJECTIVETo explore the correlation between the exposure levels and serum protein fingerprints in population exposed to silica.

METHODSLiquid chip time-of-flight mass spectrometry technology was used to investigate the serum profiles in control group (30 cases), group exposed to silica (30 cases), silicosis group (I stage, 25 cases) and suspected silicosis group (30 cases), and screen the differential expression proteins. The correlation between the levels of the differential expression proteins and the exposure levels was performed.

RESULTSFive differential expression proteins were found among 4 groups, the expression of 5081 and 5066 proteins was upregulated, and the expression of 3954, 2021 and 1777 proteins was downregulated. There was no the correlation between the exposure levels and the peak with M/Z among those proteins.

CONCLUSIONthe results of present investigation indicated there was no correlation between the exposure levels and protein/peptide peak.

Adult ; Blood Proteins ; analysis ; Case-Control Studies ; Dust ; analysis ; Female ; Humans ; Male ; Mass Spectrometry ; Middle Aged ; Occupational Exposure ; analysis ; Peptide Mapping ; Proteomics ; Silicon Dioxide ; toxicity ; Silicosis ; blood

OBJECTIVETo seek differentially expressed serum proteins in recovered SARS patients complicating avascular necrosis of femoral head (AVNFH).

METHODS2-DE and MALDI-TOF MS were used to study the comparative serum proteomics among female SARS AVNFH group, female SARS non-AVNFH group and female healthy group. ELISA method was used to detect serum amyloid P component in individual serum; specificity and sensitivity of serum amyloid P component were analyzed.

RESULTSAverage protein points on 2-DE of 3 groups were 632 +/- 28, 671 +/- 55, 688 +/- 42 respectively, and the matching rate of protein points was ranged from 85% to 95%; eighteen differentially expressed proteins were discovered including transthyretin, serpin peptidase inhibitor, alpha-1-antitrypsin precursor, serum amyloid P components, etc. Compared to healthy group and SARS non-AVNFH group, transthyretin, C4B3, fibrinogen gamma, apolipoprotein L, apolipoprotein A-IV precursor, albumin and prealbumin showed lower expression, inversely serpin peptidase inhibitor, alpha-1-antitrypsin precursor and serum amyloid P components showed higher expression in serum in the SARS AVNFH necrosis group. The serum amyloid P component in 3 groups were 0.54 +/- 0.30 ng/ml, 0.83 +/- 0.39 ng/ml, 1.21 +/- 0.29 ng/ml respectively. The areas under the ROC curve on serum amyloid P component was 0.854, the specificity was 77.8% and the sensitivity was 85.2%.

CONCLUSIONThere were differentially expressed serum proteins in three groups. Serum amyloid P components might be one of the potential biomarkers in serum of recovered SARS patients complicating avascular necrosis of femoral head.

Adult ; Blood Proteins ; analysis ; Case-Control Studies ; Electrophoresis, Gel, Two-Dimensional ; Female ; Femur Head Necrosis ; blood ; etiology ; Humans ; Proteomics ; Severe Acute Respiratory Syndrome ; blood ; complications ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective: To analyze the clinical characteristics of neonatal tracheotomy in neonatal intensive care unit (NICU). Methods: This single-center retrospective study included 14 neonates admitted to NICU of Beijing Children's Hospital, Capital Medical University from January 2016 to August 2021, and were<28 days of age on admission, who met the criteria of tracheotomy and finally completed the procedure. The clinical characteristics including age, weight, duration of ventilation, etiology of tracheotomy, length of hospital stay and prognosis were summarized and analyzed. Wilcoxon signed-rank test was used to compare the weight gain velocity and the duration of ventilation before and after tracheotomy. Paired t-test was used to compare the hospitalization length before and after tracheotomy. Spearman correlation was used to analyze the correlation between the clinical characteristics and outcomes. Results: For the 14 neonates, the gestational age was (38±4) weeks and birth weight was (2 824±949) g. Nine of them were male. The age on transportation was 16 (6, 25) d. A total of 10 neonates were on invasive ventilation on admission, the other 4 were on nasal continuous positive airway pressure support. Bilateral vocal cord paralysis (7 cases) was the commonest cause of tracheotomy. The age on operation was 33 (22, 44) d. There were statistically significant differences in duration of ventilation and weight gain velocity before and after operation (19.00 (10.50, 34.00) vs. 0.86 (0.06, 3.25) d, 1.66 (-0.16, 5.54) vs. 4.69 (2.30, 9.32) g/(kg·d), Z=3.01 and -1.98, both P<0.05). The total hospital stay in NICU was (37±12) d. One neonate died during hospitalization. The existence of pneumonia on admission was positively correlated to NICU stay length (r=0.57, P=0.027), the pre-operational weight gain velocity was negatively correlated to the post-operational NICU stay length (r=-0.73, P=0.020). There were 4 neonates de-cannulated during 7-38 months after the tracheotomy, and 5 neonates still wearing the tracheal cannulation during 15-66 months after the tracheotomy. Two neonates died and 2 neonates lost follow-up after discharge. All neonates could not vocalize normally before de-cannulation, and the language development obviously lagged behind the normal age group after de-cannulation. Conclusions: Bilateral vocal cord paralysis is the commonest cause of neonatal tracheotomy. The benefit of tracheotomy for NICU neonates with surgical indications is obvious, especially in facilitating extubation and improving weight gain.
Child ; Female ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Neonatal ; Male ; Retrospective Studies ; Tracheotomy ; Vocal Cord Paralysis ; Weight Gain

OBJECTIVETo study the effectiveness of waist circumference cut-off values in predicting the prevalence of metabolic syndrome (MetS) and risk factors in adults in China.

METHODSA cross-sectional survey was condcuted in 14 provinces (autonomous region, municipality) in China. A total of 47,325 adults aged⋝20 years were selected by multistage stratified sampling, and questionnaire survey and physical and clinical examination were conducted among them. MetS was defined according to the International Diabetes Federation (IDF) criteria and modified IDF criteria.

RESULTSThe age-standardized prevalence of MetS was 24.2% (22.1% in men and 25.8% in women) and 19.5% (22.1% in men and 18.0% in women) according to the IDF criteria and modified IDF criteria respectively. The age-standardized prevalence of pre-MetS was 8.1% (8.6% in men and 7.8% in women) according to the modified IDF criteria. The prevalence of MetS was higher in urban residents than rural residents and in northern China residents than in southern China residents. The prevalence of central obesity was about 30% in both men and women according to the ethnicity-specific cut-off values of waist circumference for central obesity (90 cm for men and 85 cm for women). Multivariate regression analysis revealed no significant difference in risk factors between the two MetS definitions.

CONCLUSIONUsing both the modified IDF criteria and ethnicity-specific cut-off values of waist circumference can provide more useful information about the prevalence of MetS in China. Conclusion Using both the modified IDF criteria and ethnicity-specific cut-off values of waist circumference can provide more useful information about the prevalence of MetS in China.

Adult ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Metabolic Syndrome ; diagnosis ; epidemiology ; Middle Aged ; Obesity ; epidemiology ; Prevalence ; Risk Assessment ; Risk Factors ; Waist Circumference