The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Autopsy ; Coxsackievirus Infections ; Enterovirus ; isolation & purification ; Enterovirus B, Human ; isolation & purification ; Enterovirus Infections ; Hand, Foot and Mouth Disease ; pathology ; virology ; Humans ; Infant ; Male

BACKGROUND: It is quite difficult to produce a decellularized lung scaffold, in which cells are removed and the extracellular matrix components (ECM) are preserved effectively. Perfusion of detergent-enzymes is an effective method with wide applications for decellularized lung scaffolds. OBJECTIVE: To investigate the effects of two detergents (sodium deoxycholate, SDC and sodium dodecyl sulfate, SDS) on the preparation of decellularized lung scaffolds. METHODS: Twenty-four male Sprague-Dawley rats were randomized into three groups: control group with no intervention, SDC group and SDS group. Decellularized lung scaffolds were prepared by perfusion of SDC or SDS combined with enzymes. The rat lung tissues in the three groups were taken for histological staining, immunofluorescent staining and DNA quantification. A549 cells were cultured and seeded onto the decellularized lung scaffolds for 7 days followed by hematoxylin-eosin staining. The decellularized lung scaffolds prepared by perfusion of SDC or SDS were subcutaneously implanted into the rat back, and the implants were retrieved and assessed by Masson staining after 2 weeks. RESULTS AND CONCLUSION: In the control group, there were abundant cells in the lung tissues. In the other two groups, the decellularized lung scaffolds were nearly transparent, and the morphology of the SDC scaffold was more close to the native lung. There were no residual cells and nuclei on the two scaffolds, and the DNA content in the SDS and SDC groups was significantly lower than that in the control group (P＜ 0.01). At 7 days of culture, A549 cells cultured on the SDS and SDC scaffolds migrated from the edge to the center of the scaffold. Comparatively speaking, the migration ability of A549 cells on the SDC scaffolds was stronger, and there was obvious cell invasion and growth in the middle part of the lung. After 2 weeks of scaffold transplantation, the SDC implants poorly fused with the surrounding tissues, with a clear boundary, a large number of infiltrating cells distributed evenly, and intravascular blood cells were clearly visible; the number of new blood vessels with larger diameter in the SDC scaffold was significantly higher than that in the SDS scaffold. These findings indicate that the SDC scaffold has better biocompatibility than the SDS scaffold, which can fuse with the surrounding tissues faster and produce more infiltrating cells and new blood vessels.

Objective To compare the antiarrhythmic effects and safety of landiolol and esmolol .Methods The sequential method was used to detect the median lethal dose ( LD50 ) and 50%effective dose ( ED50 ) of the two drugs.Treatment index were calculated by LD 50/ED50 .Several of arrhythmia models were used to study the antiarrhythmic effects of landio-lol and esmolol in the equivalent dose .Results ED50 and LD50 of landio-lol and esmolol were 14.4 , 17.3 mg? kg -1 and 347 , 100 mg? kg -1 re-spectively , TI were 24.1 and 5.8.The ratio of equivalent dose between landiolol and esmolol on anti -chloroform induced arrhythmia was 0.83∶1.Landiolol and esmolol have similar anti -arrythmia effect on va-rious arrhythmia models.Esmolol has a stronger inhibition function on the heart rate than that of landiolol .Conclusion Compared with esmolol , landiolol has similar antiarrhythmic effect , gentle and slow effect on heart rate, higher treatment index and safety in equivalent dose .

No abstract available.
Aged, 80 and over ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/*genetics ; China ; Drug Resistance, Multiple, Bacterial ; Humans ; Male ; Methyltransferases/*genetics ; Microbial Sensitivity Tests ; RNA, Ribosomal, 16S/genetics/metabolism ; Serratia marcescens/drug effects/*enzymology/*genetics/isolation & purification ; Urinary Tract Infections/diagnosis/microbiology

OBJECTIVETo compare the efficacy and safety of tolterodine and oxybutynin in the treatment of idiopathic overactive bladder in children.

METHODSA total of 204 children with idiopathic overactive bladder were randomly divided into three groups (n=68 each): placebo, tolterodine-treated and oxybutynin-treated. The efficacy and safety were evaluated two weeks after treatment.

RESULTSThe effective rate was 25% in the placebo group, 89% in the tolterodine-treated group, and 92% in the oxybutynin-treated group. The effective rate in the two treatment groups was significantly higher than that in the placebo group (P<0.05). There was a similar efficacy between the two treatment groups. The incidence of adverse events in the tolterodine-treated group (28%) was significantly lower than that in the oxybutnin-treated group (57%) (P<0.05).

CONCLUSIONSTolterodine has a similar efficacy to oxybutynin in the treatment of idiopathic overactive bladder in children, with better safety in pharmacotherapy.

Adolescent ; Benzhydryl Compounds ; adverse effects ; therapeutic use ; Child ; Child, Preschool ; Cresols ; adverse effects ; therapeutic use ; Female ; Humans ; Male ; Mandelic Acids ; adverse effects ; therapeutic use ; Muscarinic Antagonists ; therapeutic use ; Phenylpropanolamine ; adverse effects ; therapeutic use ; Tolterodine Tartrate ; Urinary Bladder, Overactive ; drug therapy

Objective To explore the diagnosis and treatment of eosinophilic cystitis (EC) in children.Methods Six patients who experienced EC in Nanjing Children's Hospital Affiliated to Nanjing Medical University from Mar.2007 to Feb.2012 were analyzed and the relevant literature was reviewed.This group included 5 male and 1 female,with mean age of 5.3 years.Their major complains included urinary frequency in 5 cases,odynuria in 3 cases,dysuria in 2 cases,hematuria in 2 cases,proteinuria in 2 cases,urinary dribbling in 1 case,and nocturia in 1 case.Physical examination and urine culture was positive in 1 case,eosinophil proportion of peripheral blood increased in 3 cases,imaging studies suggested the bladder wall thickening in 5 cases and space-occupying lesions in 2 cases.Cystoscopy in 4 cases and biopsy in 3 cases showed EC.Results Three patients were cured after receiving anti-infection and urine alkalinization,and another 3 patients were cured after adding Prednisone and Ketotifen,or Oxybutynin or Loratadine.Five patients were followed up for 1-60 months,and clinical symptoms,blood,urine,and B-ultrasound in these patients were normal.The information of 1 case was lost.Conclusions The incidence of EC in children tends to increase in recent years,the diagnosis can be confirmed combined with the clinical manifestations,laboratory tests,imaging studies and histological examination.Conservative treatment is preferred.

Objective To describe a simple and efficient method to obtain large quantities of higllly purified Schwann cells(SCs).Methods SCs were isolated from the sciatic and brachial nerves of 3-to 5-day-old newborn SD rats by collagenase digestion.The isolated SCs were plated at the density of3x105/mL for primary culture,several rounds oftrypsin digestion were performed within 72 h to purify SCs. Results Compared with the purification using 1.25 g/L trypsin digestion in serial differential detachment procedures, Our protocol allowed easier and more efficient separation of the SCs from the fibroblasts.Immunocytochemical staining showed that the purity of the SCs exceeded 95%.Conclusion The purification protocol of the SCs we established can be easily carried out and yields well reproducible results to obtain large quantities ofhighly purified SCs for transplantation studies.

Objective To construct a knowledge graph of Hai Yao Ben Cao(Oversea Materia Medica)based on Neo4j technology,and to analyze the knowledge graph and discuss its significance.Methods Taking the original text of Hai Yao Ben Cao as the data source,the data information related to Chinese medicine was extracted,and then the Excel data table was established.The data information included a total of 13 items,and covered the serial number of Chinese medicines,name of Chinese medicines,medicinal part,origin,medicinal property,medicinal flavor,efficacy,toxicity,and various indicators.The extracted information was preprocessed and standardized.And then the constructed knowledge graph was stored and analyzed by graph database Neo4j.Results The knowledge graph of Hai Yao Ben Cao based on Neo4j was constructed,and a total of 518 entities and 1 046 entity relationships were obtained,which were mainly related with origin,medicinal flavor,and efficacy.The results of the entity statistics presented the status of medicinal development and the distribution of oversea medicines in Late Tang and Five Dynasties.The constructed knowledge graph of Hai Yao Ben Cao ensured the realization of information query and content visualization.Conclusion The constructed knowledge graph of Hai Yao Ben Cao intuitively shows the relationship among the category,origin,medicinal property,medicinal flavor,toxicity and efficacy of the medicines in the book,which will lay a foundation for further research on the medicine-medicine relationship and medicine-disease relationship,and for the study of the history of traditional Chinese medicine(TCM)development.The knowledge graph of Hai Yao Ben Cao will also provide methods for the rational and effective utilization of ancient TCM books,being beneficial to the modernization of TCM.

OBJECTIVETo investigate the association of polymorphisms of CDT1 and GMNN gene, two important genes participating in DNA replication, with the risk of sporadic breast cancer.

METHODSUsing polymerase chain reaction-restriction fragment length polymorphism (PCR - RFLP) and the primer-introduced restriction analysis (PIRA)-PCR assay to genotype the CDT1 838G/A and GMNN 387C/A polymorphisms in a case-control study of 427 breast cancer cases and 477 cancer-free controls in a Chinese population.

RESULTSNo significant association of the CDT1 838G/A and GMNN 387C/A polymorphisms with the risk of breast cancer was found (adjusted OR:1.16, 95% CI:0.88-1.54 for CDT1 GA+AA genotypes and adjusted OR:0.90, 95% CI:0.67-1.21 for GMNN CA+AA genotypes). However, in the stratified analyses, a significant association of CDT1 GA+AA genotypes with breast cancer risk among subjects with family history of cancer was found (adjusted OR:2.21, 95% CI:1.20-4.09).

CONCLUSIONThese findings suggest that the CDT1 838G/A and GMNN 387C/A polymorphisms may not play a major role in the etiology of breast cancer, but CDT1 variant may have a potential role only in genetically susceptible women.

Adult ; Asian Continental Ancestry Group ; genetics ; Breast Neoplasms ; ethnology ; genetics ; Case-Control Studies ; Cell Cycle Proteins ; genetics ; China ; Female ; Geminin ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length