OBJECTIVETo express the fusion protein of glutathione S-transferase (GST) and human Id-2 in E. coli and prepare the polyclonal antibodies against Id-2.

METHODSThe coding sequence of Id-2 gene was amplified by RT-PCR from the total RNA of breast cancer tissue. The recombinant plasmid was identified by PCR, restriction endonuclease digestion analysis and sequencing. The fusion protein GST-Id-2 expressed in E. coli following IPTG induction was purified by glutathione-agarose affinity chromatography and used to immunize rabbits to prepare the polyclonal antibodies against GST-Id-2.

RESULTSPCR, restriction endonuclease digestion and sequence analyses showed that the Id-2 gene had been correctly inserted into pGEX-6P-1 vector, and the GST-Id-2 fusion protein expressed had a relative molecular mass of approximately 40,000 as shown by SDS-PAGE. The polyclonal antibodies obtained from the rabbit sera were found to specifically react with purified Id-2 by Western blotting, ELISA and agar gel immunodiffusion (AGP).

CONCLUSIONThe prepared polyclonal antibodies against Id-2 allow effective Id-2 detection and facilitate further investigation of the structure and antigen epitope of Id-2.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Breast Neoplasms ; genetics ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Immune Sera ; biosynthesis ; Inhibitor of Differentiation Protein 2 ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo construct the anti-lung tumor gene differentially expressed bank of wild mouse and to explore the mechanisms of the TW wild mouse suppressing the occurring of lung tumor.

METHODSUsing suppression subtractive hybridization (SSH) technique, the differentially expressed genes between TAF1 mouse and A/wy mouse were selected out and the subtracted cDNA bank was constructed. 166 clones were performed DNA sequencing and then were assayed by blast programme.

RESULTSAmong the blast results of 166 differentially expressed clones, 87 known genes (mRNA or cDNA) were in homology with 134 clones and were divided into 7 classifications according to the biological role.14 DNA fragments were in homology with 32 clones, in which 20 clones were in homology with 9 mouse DNA sequences, 2 clones were in homology with one bacterial gene sequences and 3 clones were clone vector.

CONCLUSIONWith SSH technique, the anti-lung tumor gene differentially expressed bank of wild mouse are successfully constructed.

Animals ; DNA, Complementary ; genetics ; Female ; Gene Expression Profiling ; Gene Library ; Lung Neoplasms ; genetics ; Male ; Mice ; Mice, Inbred Strains ; genetics ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Tumor Cells, Cultured

OBJECTIVECatheter closure of perimembrane ventricular septal defect (PMVSD) using the Amplatzer asymmetric ventricular septal defect occluder (AAVSDO) is a potential alternative for open surgical repair. However, the profile of the device obtained after closure probably continues to change some concerns regarding its safety. This study was designed to evaluate the morphologic changes of AAVSDO by transthoracic echocardiography examination (TTE) and X-ray examination after transcatheter closure of PMVSD.

METHODSA total of 23 patients, aged 2.5 - 47.0 years, with PMVSDs underwent transcatheter closure with the AAVSDO. Each patient underwent TTE and X-ray examination with same radiography system immediately, 72 hours, 6 months and 1 year after the transcatheter closure procedures, respectively. Seven parameters were measured by TTE and X-ray to evaluate the morphologic changes of AAVSDO: the distance between superior edge of left disk of AAVSDO and aortic valve (L(1)), the distance between superior extremity edge of two disks of the occluder (L(2)), the distance between inferior extremity edge of two disks of the occluder (L(3)), the distance between two marks on the left and right disks (L(4)), the diameter of left disk (D(1)), the diameter of left disk (D(2)), and the degree of angle between left and right disks (alpha). In the meantime, the influences of occluder's morphologic changes were evaluated by TTE.

RESULTSThe PMVSD diameter ranged from (8.53 +/- 4.82) mm (3.91 - 17.0 mm). The device diameter ranged from 6 - 18 (10.34 +/- 7.16) mm. AAVSDO was performed successfully in all the 23 patients who underwent immediately, 72 hours and 6 months follow-up after catheter closure, and 12 patients accomplished 1-year follow-up. L(2), L(4) and alpha were shorter at 6-months than 72-hours after procedures, but D(1) and D(2) were greater at 6-months than 72-hours after procedures in all patients. Alpha was decreased further 1 year after the procedure, but other parameters did not change significantly compared with those at 6-months after procedures. The lower profile and smaller thickness of AAVSDO were observed in the follow-up. Meanwhile, D(1) and D(2) changed significantly during the follow-up. L(4) had a weak positive correlation with device size (r = 0.47, 0.33 and 0.39, respectively) and with alpha (r = 0.47, 0.53 and 0.49, respectively), and had weak negative correlation with D1 (r = -0.27, -0.45 and -0.29, respectively). After deployment of the prosthesis there was no residual shunt in 20 of 23 patients (87%). There was a trivial residual shunt that disappeared at the six month follow up in two patients, and a small residual shunt that disappeared at the one year follow up in one patient. The morphologic changes of AAVSDO did not influence the structure near PMVSD.

CONCLUSIONThe morphologic changes of Amplatzer occluder were observed in the follow-up. The change of the occluder was safe and beneficial to improve short-term curative effect.

Adolescent ; Adult ; Cardiac Catheterization ; Child ; Child, Preschool ; Follow-Up Studies ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Middle Aged ; Prosthesis Implantation ; instrumentation ; Radiography ; Septal Occluder Device ; Ultrasonography ; Young Adult

OBJECTIVETo evaluate the accuracy and practicability of detecting viable myocardium by CARTO voltage mapping in swine model of acute myocardial infarction (MI).

METHODSMI was induced in 13 anesthetized swines via occluding the distal of left anterior descending coronary arteries by angioplasty balloon for 60-90 minutes. The viable myocardium detection by CARTO voltage mapping was made after reconstruction of the left ventricle using CARTO and the results were compared with TTC staining. The standard of CARTO voltage to detect viable myocardium was 0.5 - 1.5 mV while viable myocardium showed pink color by TTC staining.

RESULTSEleven out of 13 swines survived the operation and 2 swines died of ventricular fibrillation at 45 and 65 minutes post ischemia. Left ventricle was divided into 16 segments and 176 segments from 11 swines were analyzed. Viable myocardium detected by CARTO voltage mapping was identical as identified by TTC staining (Kappa = 0.816, P < 0.001). Taken the TTC result as standard, the sensitivity, specificity and accuracy rate of CARTO voltage mapping are 71.8%, 96.5% and 90.9% respectively.

CONCLUSIONCARTO voltage mapping could be used as a reliable tool to detect viable myocardium in this model.

Animals ; Cell Survival ; Disease Models, Animal ; Electrophysiologic Techniques, Cardiac ; Female ; Male ; Myocardial Infarction ; physiopathology ; Myocardium ; cytology ; Swine

OBJECTIVETo investigate the therapeutic effects of upper airway stenosis after Le Fort III osteotomy and midfacial distraction osteogenesis (DO).

METHODSEleven cases (age, 5-16 yrs) with severe midface dysostosis complicated with exophthalmos, anterior crossbite and upper airway stenosis were treated by using Le Fort III osteotomy and midfacial DO from August 2000 to February 2007. The 3D reconstruction of the upper-airway from CT data was used to evaluate the upper airway volume before and after the operation. And meanwhile polysomnography was carried out to demonstrate the upper airway functional changes.

RESULTSThere was a 64.3% mean increase [mean, (9.13 +/- 6.94) ml, P < 0.05] in upper airway volume in the 11 cases after the operations. It showed that there was significant improvements in the indexes of polysomnography after the operations, such as apnea and hypopnea index, average SaO2, minimum oxygen saturation and snore index.

CONCLUSIONSThe Le Fort III osteotomy and midfacial distraction osteogenesis can efficiently relieve the symptoms of upper-airway stenosis in severe midfacial dysostosis.

Acrocephalosyndactylia ; complications ; Adolescent ; Airway Obstruction ; etiology ; surgery ; Child ; Child, Preschool ; Craniofacial Dysostosis ; complications ; Female ; Follow-Up Studies ; Humans ; Male ; Osteogenesis, Distraction ; methods ; Osteotomy, Le Fort ; Treatment Outcome

OBJECTIVETo find a better method for harvesting highly purified neurons by comparing three methods used for co-culture of neurons and astrocytes.

METHODSThe co-culture models of neurons and astrocytes were established by primary culture, Banker's co-culture method or Transwell cell-culture inserts. The neurons and astrocytes cultured in vitro were from neonatal rats.

RESULTSThe highly purified neurons were not harvested by primary culture because the neurons and astrocytes grew on the same cover slip and it was difficult to control the growth velocity of astrocytes. The highly purified neurons were harvested by Banker's co-culture method or the method using Transwell cell-culture inserts, but the procedure of the former was more complicated than that of the later.

CONCLUSIONSThe culture method using Transwell cell-culture inserts is recommended for the establishment of the co-culture system of neurons and astrocytes.

Animals ; Astrocytes ; Cells, Cultured ; Coculture Techniques ; Neurons ; cytology

OBJECTIVETo investigate the phenotype, frequency and function of CD4+ T cell subsets and the relevant cytokines, as well as the relationship between these cells and appearance of pneumonia of novel (H1N1) influenza A patients.

METHODS68 healthy people, 53 confirmed novel A(H1N1) influenza patients without pneumonia and 16 confirmed severe novel A (H1N1) influenza patients with pneumonia were enrolled in this study. Viral load in nasopharyngeal swabs specimens was measured by real time PCR assay. The phenotype and percentage of CD4+ T cell subsets including Th1, Th2, Th17, and Treg cells were measured by Flow cytometry analysis. The relevant cytokines in plasma including TGF-beta, IL-6 and IFN-gamma were measured by ELISA. Data was analyzed by one way ANOVA.

RESULTSIt was found that peak viral load and viral shedding period of severe patients with pneumonia was significantly increased compared with mild patients without pneumonia (P < 0.05). The percentage of Th17 cells of severe patients with pneumonia was significantly diminished compared to that of healthy subjects and mild patients without pneumonia (P < 0.05). However, Th1, Th2, Treg cells frequencies had no significant differences (P > 0.05) among these three groups. The level of TGF-beta in plasma for the severe patients with pneumonia was also significantly decreased compared to that of healthy subject and mild patients without pneumonia (P < 0.05). The viral shedding period inversely correlated with the frequency of Th17 cells (r = - 0.38, P < 0.05).

CONCLUSIONH1N1 influenza A virus can inhibit Th17 cells to differentiate, particularly more extent in patients with pneumonia. Impaired Th17 cells may correlate with viral clearance and pneumonia of novel H1N1 influenza A patients.

Adolescent ; Adult ; CD4-Positive T-Lymphocytes ; immunology ; Cytokines ; immunology ; Female ; Humans ; Immunity, Cellular ; immunology ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza, Human ; immunology ; Male ; Pneumonia, Viral ; immunology ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo investigate the genetic structure of X chromosome in Mongolia, Ewenki, Elunchun and Dawoer in Inner Mongolia.

METHODSNine short tandem repeat (STR) markers on the X chromosome (DXS6789, DXS101, DXS8378, DXS7132, DXS7133, DXS7423, DXS6804, DXS6799 and HPRTB) were analyzed in the four populations from Inner Mongolian (Mongol, Ewenki,Oroqen and Daur) for their genetic diversity, forensic suitability and possible genetic affinities of the populations. Frequencies and other parameters of forensic interest were computed.

RESULTSThe results revealed that the nine markers described here have a moderate degree of variability in the population groups. And there are significant differences in the genetic variability among the populations. Genetic distance and cluster analyses show very low genetic distance between Mongol and Han (Xi'an) communities. The results based on genetic distance analyses are consistent with earlier studies based on linguistic as well as immigration history and origin of these populations.

CONCLUSIONThe nine STR loci studied here were found not only useful in studying genetic variations between populations but also suitable for human identity testing.

China ; ethnology ; Chromosomes, Human, X ; genetics ; Cluster Analysis ; Ethnic Groups ; genetics ; Female ; Genetic Variation ; Humans ; Male ; Microsatellite Repeats ; genetics

Objective:To compare the predictive value of the HEART, TIMI and GRACE scores for major adversecardiovascular events (MACEs) at 7 and 28 days in patients with actue non-ST-segment elevation myocardial infarction (NSTEMI).Methods:More than 12 000 patients with chest pain from the Emergency Department of Zhongshan Hospital Affiliated to Fudan University from October 2017 to October 2018 were studied, including 566 patients with cardiogenic chest pain, 105 patients with ST-segment elevation myocardial infarction (STEMI) excluded and 15 patients lost to follow-up. Finally, 109 patients with NSTEMI and 337 non-myocardial patients with cardiogenic chest pain were enrolled. NSTEMI patients were divided into subgroups according to whether MACEs occurred. LSD t-test, Mann-Whitney U test or χ2 test were used to analyze and compare the differences between the two subgroups about the baseline data, clinical data, HEART, TIMI and GRACE scores at the time of visit. Multivariate logistic regression analysis was used to explore the independent factors of MACEs at 7 and 28 days. And the predictive values of different scores for 7-day MACEs and 28-day MACEs were compared in NSTEMI patients through the receiver operating characteristic (ROC) curve. Results:Compared NSTEMI patients with non-myocardial patients with cardiogenic chest pain, we found a statistically significant differences in sex, past history of coronary heart disease,≥3 risk factors for atherosclerosis, electrocardiogram, high-sensitivity troponin T (hs-cTnT), creatinine value, past history of myocardial infarction, HEART score, TIMI score and GRACE score. In further subgroup analysis of NSTEMI patients who were divided according to whether MACEs occurred, we found previous history of stroke and increased hs-cTnT were statistically different in 7 days after the onset of the disease. The multivariate analysis showed that the previous history of stroke and increased hs-cTnT were independent factors for the occurrence of MACEs at 7 days after the onset of NSTEMI; The previous history of stroke and increased hs-cTnT, electrocardiogram ST segment depression and TIMI score were statistically different at 28 days after the onset of NSTEMI. The multivariate analysis showed that the previous history of stroke and TIMI score were independent factors for the occurrence of MACEs at 28 days after the onset of NSTEMI patients. ROC curve indicated that the predictive value of TIMI score (AUC=0.715, 95% CI: 0.482-0.948) was better than HEART (AUC=0.659, 95% CI: 0.414-0.904) and GRACE scores (AUC=0.587, 95% CI: 0.341-0.833)in predicting MACEs in NSTEMI patients. Conclusions:HEART score, TIMI score and GRACE score can be used to evaluate NSTEMI patients. There is an independent predictive value on TIMI score for the occurrence of 28-day MACEs in NSTEMI patients.

Objective: To summarize the research hotspots and knowledge structure of pediatric teaching model by introducing the co-word clustering analysis into the research on the current status of pediatric teaching model and conducting a visualized analysis of related articles in the past decade, and to provide a reference for further development of pediatric teaching model. Methods: In July 2017, CNKI and Wanfang Data were searched for related articles published from January 2006 to June 2017, with "pediatrics" and "teaching model" as the key words. The articles which met the research criteria were screened out manually, and Word and Bicomb 2.0 were used to identify high-frequency key words. SPSS 20.0 was used to plot the diagram of the cluster analysis, and Ucinet 6.0 and Newdraw were used to generate the co-occurrence network map to show the relationship between high-frequency key words. Results: A total of 367 articles were included and 31 high-frequency key words were extracted, among which the most frequently used key word was "PBL teaching method". Studies in this field were roughly classified into four types, i.e., teaching method in pediatric practice/and clerkship, contents of cultivation and assessments in pediatric teaching, teaching methods for pediatric nursing, and objects of the reform of teaching methods. The key words of "PBL teaching method", "multimedia teaching", "case-based teaching", and "nursing" were located at the center of the co-occurrence network map. Conclusion Exploration and practice of various teaching methods have been the main research topics in pediatric teaching model in China over the past decade. PBL is a research hotspot in this field and is often combined with multimedia, case teaching, or other methods in pediatric teaching. In addition, the application of evidence-based medicine has attracted more and more attention.