Objective To investigate the changes of cardiac function at pre -and post -treatment in preterm infants with patent ductus arteriosus (PDA)in order to guide drug treatment.Methods Totally 84 preterm infants with PDA admitted to Neonatal Intensive Care Unit of Xuzhou Hospital Affiliated to Medical College of Southeast University from July 201 2 to June 201 4 were divided into 4 groups according to treatment drug:Ibuprofen group (27 cases),Indo-methacin group (24 cases),control group (1 1 cases),and Paracetamol group (22 cases).Patients were also divided into symptomatic PDA group (38 cases)and asymptomatic PDA group (46 cases)according to severity;PDA closed group (69 cases)and PDA unclosed group (1 5 cases)according to sequel.The level of plasma brain natriuretic pep-tide (BNP),cardiac troponin I (cTnI),correct QT intervals dispersion (QTcd)were monitored pre -and post -treat-ment.Data were analyzed by using SPSS 1 9.0 software.Results Three cardiac markers at post -treatment were of no significant difference among 4 treatment drugs.The changes of the cTnI and QTcd at pre -and post -treatment were of no significance.The level of BNP in symptomatic PDA group was significantly higher than that in asymptomatic PDA group at pre -treatment [(378 ±94)ng/L vs (1 47 ±75)ng/L,t =2.584,P =0.01 4].In the symptomatic PDA group,the level of BNP at post -treatment [(1 82 ±81 )ng/L]was significantly decreased than that at per -treatment (t =2.741 ,P =0.009).In the asymptomatic PDA group,there was no significant difference between the pre - and post -treatment [(1 21 ±61 )ng/L]in the level of BNP (t =1 .254,P =0.207).There was no significant difference in the level of BNP at per -treatment between PDA closed group and PDA unclosed group [(274 ±91 )ng/L vs (289 ± 87)ng/L,t =-0.874,P =0.391 ].In PDA closed group,the level of BNP at post -treatment [(1 21 ±74)ng/L] was significantly decreased compared with that at per -treatment (t =3.580,P =0.000).In PDA unclosed group, there was no significant difference between the pre - and post -treatment [(245 ±74)ng/L]in the level of BNP (t =0.854,P =0.392).Conclusion Early medication intervention for symptomatic PDA of preterm infants is benefi-cial for the closure of PDA and for attenuating negative effects on cardiac function of PDA.

An recombinant vector pCSV-EPO for expression of EPO cDNA in mammalian cells was constructed by the techniques of gene recombinant, PCR amplification and region-specific mutagenesis. The pCSV-EPO was introduced into COS7 cells by DEAE-dextran-mediated transfection. The expression of the EPO was demonstrated by EPO-ELISA assay. At 48h post transfection, the EPO level was 25ng/ml and 72 h was 17ng/ml.

Objective To investigate the translation and rotation of healthy elbow joints during active flexion and extension, so as to provide references for the treatment of elbow instability and injuries. Methods Ten healthy subjects with no history of upper extremity trauma were recruited. Dual fluoroscopic imaging system (DFIS) was applied to quantify six-degree-of-freedom (6-DOF) kinematics of humeroulnar and humeroradial joint from full extension to maximum flexion in supination position. Correlation analysis was used to evaluate the relationship between the 6-DOF kinematics of the elbow joint. Results Valgus angle of humeroulnar joint gradually decreased from 15.2°±3.1° to 5.3°±2.3° with the flexion increasing. Valgus angle of humeroradical joint gradually decreased from 19.7°±4.2° to 8.2°±2.4° from full extension to maximum flexion. The valgus angle of humeroulnar and humeroradial joint was linearly related to the flexion angle during flexion and extension. The internal rotation of humeroulnar joint was quadratic nonlinearly related to the flexion angle during motion. The maximum internal rotation of the ulnar was 4.0°±4.9° at 110° flexion, and the maximum external rotation was 5.1°±4.2°. The internal rotation of humeroulnar joint gradually increased from 3.2°±16.0° to 27.2°±18.0°. Conclusions During normal flexion and extension of the elbow, the valgus angle of humeroulnar joint decreased linearly, while the internal and external rotation angle showed a nonlinear change, which first rotated internally and then rotated externally. The valgus angle of humeroradial joint decreased linearly and the internal rotation angle increased linearly. Therefore, humeroulnar joint is not a hinge joint during elbow flexion and extension. There is a kinematic difference between humeroulnar joint and humeroracial joint. For clinical treatment of complex elbow injuries, elbow instability and elbow replacement, different motion characteristics in joints should be considered to improve the outcome after surgery.

Blast injury of the chest injury is the most common wound in modern war trauma and terrorist attacks, and is also the most fatal type of whole body explosion injury. Most patients with severe blast injury of the chest die in the early stage before hospitalization or during transportation, so first aid is critically important. At present, there exist widespread problems such as non-standard treatment and large difference in curative effect, while there lacks clinical treatment standards for blast injury of the chest. According to the principles of scientificity, practicality and advancement, the Trauma Society of Chinese Medical Association has formulated the guidance of classification, pre-hospital first aid, in-hospital treatment and major injury management strategies for blast injury of the chest, aiming to provide reference for clinical diagnosis and treatment.

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Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the SARS-CoV. It is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving the SP Group (Singapore) more distant. This result appears to suggest a possible transmission path from Guangdong to Beijing/Hong Kong, then to other countries and regions.
Genome, Viral ; Haplotypes ; Humans ; Mutation ; Open Reading Frames ; Phylogeny ; SARS Virus ; genetics

We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence ; China ; Cluster Analysis ; Gene Components ; Genetic Variation ; Genome, Viral ; Genotype ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; genetics

Gene expression labeling and conditional manipulation of gene function are important for elaborate dissection of gene function. However, contemporary generation of pairwise dual-function knockin alleles to achieve both conditional and geno-tagging effects with a single donor has not been reported. Here we first developed a strategy based on a flipping donor named FoRe to generate conditional knockout alleles coupled with fluorescent allele-labeling through NHEJ-mediated unidirectional targeted insertion in zebrafish facilitated by the CRISPR/Cas system. We demonstrated the feasibility of this strategy at sox10 and isl1 loci, and successfully achieved Cre-induced conditional knockout of target gene function and simultaneous switch of the fluorescent reporter, allowing generation of genetic mosaics for lineage tracing. We then improved the donor design enabling efficient one-step bidirectional knockin to generate paired positive and negative conditional alleles, both tagged with two different fluorescent reporters. By introducing Cre recombinase, these alleles could be used to achieve both conditional knockout and conditional gene restoration in parallel; furthermore, differential fluorescent labeling of the positive and negative alleles enables simple, early and efficient real-time discrimination of individual live embryos bearing different genotypes prior to the emergence of morphologically visible phenotypes. We named our improved donor as Bi-FoRe and demonstrated its feasibility at the sox10 locus. Furthermore, we eliminated the undesirable bacterial backbone in the donor using minicircle DNA technology. Our system could easily be expanded for other applications or to other organisms, and coupling fluorescent labeling of gene expression and conditional manipulation of gene function will provide unique opportunities to fully reveal the power of emerging single-cell sequencing technologies.
Alleles ; Animals ; CRISPR-Cas Systems ; DNA End-Joining Repair ; DNA, Circular/metabolism* ; Embryo, Nonmammalian ; Gene Editing/methods* ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Genes, Reporter ; Genetic Loci ; Genotyping Techniques ; Green Fluorescent Proteins/metabolism* ; Integrases/metabolism* ; Luminescent Proteins/metabolism* ; Mutagenesis, Insertional ; Single-Cell Analysis ; Zebrafish/metabolism*

Mammals exhibit limited heart regeneration ability, which can lead to heart failure after myocardial infarction. In contrast, zebrafish exhibit remarkable cardiac regeneration capacity. Several cell types and signaling pathways have been reported to participate in this process. However, a comprehensive analysis of how different cells and signals interact and coordinate to regulate cardiac regeneration is unavailable. We collected major cardiac cell types from zebrafish and performed high-precision single-cell transcriptome analyses during both development and post-injury regeneration. We revealed the cellular heterogeneity as well as the molecular progress of cardiomyocytes during these processes, and identified a subtype of atrial cardiomyocyte exhibiting a stem-like state which may transdifferentiate into ventricular cardiomyocytes during regeneration. Furthermore, we identified a regeneration-induced cell (RIC) population in the epicardium-derived cells (EPDC), and demonstrated Angiopoietin 4 (Angpt4) as a specific regulator of heart regeneration. angpt4 expression is specifically and transiently activated in RIC, which initiates a signaling cascade from EPDC to endocardium through the Tie2-MAPK pathway, and further induces activation of cathepsin K in cardiomyocytes through RA signaling. Loss of angpt4 leads to defects in scar tissue resolution and cardiomyocyte proliferation, while overexpression of angpt4 accelerates regeneration. Furthermore, we found that ANGPT4 could enhance proliferation of neonatal rat cardiomyocytes, and promote cardiac repair in mice after myocardial infarction, indicating that the function of Angpt4 is conserved in mammals. Our study provides a mechanistic understanding of heart regeneration at single-cell precision, identifies Angpt4 as a key regulator of cardiomyocyte proliferation and regeneration, and offers a novel therapeutic target for improved recovery after human heart injuries.
Humans ; Mice ; Rats ; Cell Proliferation ; Heart/physiology* ; Mammals ; Myocardial Infarction/metabolism* ; Myocytes, Cardiac/metabolism* ; Pericardium/metabolism* ; Single-Cell Analysis ; Zebrafish/metabolism*