Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain,seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI,which contained the full-length cDNA of the NDV ZJI strain.The pNDV/ZJI,with three helper plasmids,pCIneoNP,pCIneoP and pCIneoL,were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase.After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock,an infectious NDV ZJI strain was successfully rescued.Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP.After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells,the recombinant NDV,NDV/ZJIGFP,was rescued.Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection,indicating that the GFP gene was expressed at a relatively high level.NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route.Four days post-infection,strong green fluorescence could be detected in the kidneys and tracheae,indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.

Objective To summarize clinical characteristics of female patients with delayed cardiac tamponade after percutaneous coronary intervention (PCI).Methods Materiais on clinical characteristics of 1009 elderly patients who might have delayed cardiac tamponade after PCI from Aug 1999 to May 2011 was analyzed retrospectively.Results Two patients who had delayed cardiac tamponade after PCI were female,suggesting that elderly female patients with multi-vessel coronary disease and complex coronary lesions should be monitored closely after PCI.Bedside ultrasound should be performed promptly within 6 hours after operation for unexplained low pressure in case of delayed cardiac tamponade.Two patients were recovered and discharged after pericardiocentesis and appropriate treatment.Conclusions Elderly female patients with multi-vessel coronary disease and complex coronary lesions tend to have delayed cardiac tamponade after PCI easily,whose early clinical characteristics should be noted and evaluated.

OBJECTIVETo explore the clinical classification and ideal therapy for maxillofacial AVMs.

METHODSAccording to the clinical characteristics, 106 patients with maxillofacial AVMs were divided into the 4 types Of them, 38 cases were cystic dilatation lesions, 22 cases were limited thicken lesions, 42 case were diffuse thicken lesions, 4 cases were central maxillary hemangioma. 106 patients with maxillofacial AVMs were treated in our hospital, of them, 8 cases received operation (group 1); 23 cases received embolization of supplying artery alone (group 2); 37 cases received embolization of supplying artery plus hardener intra-tumorous injection (group 3); 38 cases received embolization of supplying artery plus tumor resection (group 4).

RESULTSOf all the patients were followed up 1 - 11 years, In group 1, 2, 3, and 4, the cure rates is 62.50%, 17.39%, 89.19%, and 97.37% respectively. one patient died of embolization of abnormal communication branches between external carotid and intra-cranical arteries.

CONCLUSIONS(1) This new clinical classification is beneficial for selecting method of treatment. (2) It is necessary that a good digital subtraction angiography for maxillofacial AVMs. (3) The embolization of tumor supplying artery alone could cure the small AVM with single branch terminal blood supply. (4) The embolization of supplying artery plus hardener intratumorous injection or the embolization of supplying artery plus tumor resection is an effective method for maxillofacial AVMs.

Adolescent ; Adult ; Angiography, Digital Subtraction ; Arteriovenous Malformations ; diagnosis ; therapy ; Carotid Artery, External ; diagnostic imaging ; Child ; Child, Preschool ; Embolization, Therapeutic ; methods ; Female ; Follow-Up Studies ; Humans ; Jaw ; blood supply ; Male ; Middle Aged ; Mouth ; blood supply ; Sclerotherapy

OBJECTIVETo construct a eukaryotic expression vector containing aldehyde dehydrogenase-2 (ALDH2) gene and investigate the effects and its possible mechanisms of ALDH2 gene on cell proliferation and anti-oxidative damage in the K562 cells.

METHODSAn eukaryotic expression vector containing the ALDH2 gene cloned from human hepatocytes was constructed and transfected into K562 cells by liposome. RT-PCR and Western blot were used to evaluate the expression of ALDH2. MTT assay was used to check the cell proliferation and trypan blue exclusion to check K562 cells damage induced by hydrogen peroxide (H2O2). RT-PCR and fluorescence spectrophotometry were used to determine the expression of heme oxygenase-1 (HO-1) and the generation of intracellular reactive oxygen species (ROS) respectively.

RESULTSRT-PCR and Western blot analysis showed distinct higher ALDH2 protein expression in gene transfected group. The latter group had a higher cell proliferation (P < 0.05) and survival rate against H2O2 induced-oxidative damage, being increased by 7.8 times (IC(50) was 12.3 µmol/L and 1.4 µmol/L for K562-pcDNA3.1-ALDH2 and control cells, respectively, P < 0.01). The HO-1 mRNA expression and the generation of intracellular ROS were downregulated at a specific concentration of H2O2 in the ALDH2 gene transfected group.

CONCLUSIONALDH2 gene transfection can protect K562 cells against oxidative damage, and the downregulation of HO-1 expression and intracellular ROS may be involved in this process.

Aldehyde Dehydrogenase ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Hydrogen Peroxide ; K562 Cells ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo investigate the association between the exons 2 to 4 of the MICA gene and ankylosing spondylitis (AS).

METHODSBy PCR-SSOP, DNA samples from 56 AS patients and 112 random healthy individuals, as normal control were genotyped to analyse the polymorphism in exons 2, 3, 4 of the MICA alleles.

RESULTSMICA*008 was dominant in MICA allele,accounted for 32.14% and 30.36% in AS patients and normal controls respectively. The frequency of MICA*007 was significantly increased in AS patients, when compared with normal controls (chi-square=10.18, P<0.05, RR=2.50). No difference was found in the other MICA alleles. The haplotype analysis revealed that there were the strong linkage disequilibrium between MICA and HLA-B of AS patients, and normal controls. There was a difference in MICA*007-B27 between two groups (chi-square=18.46, P<0.05, RR=7.47). Both HLA-B27 and MICA*007 were strongly associated with AS. Stratified analysis showed that HLA-B27 was significantly relative to AS,while it was not found between MICA*007 and AS.

CONCLUSIONThe increased frequency of MICA alleles may be due to its strong linkage disequilibrium with HLA-B27.

Alleles ; Exons ; genetics ; Gene Frequency ; Genotype ; HLA-B Antigens ; genetics ; HLA-B27 Antigen ; genetics ; Histocompatibility Antigens Class I ; genetics ; Humans ; Linkage Disequilibrium ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Spondylitis, Ankylosing ; genetics

OBJECTIVETo investigate the inhibitory effects of CaMKIIN on acute myeloid leukemia cell line HL-60 to explore a novel therapeutic target of leukemia.

METHODSHuman CaMK II N gene expression vector pcDNA3.1/hCaMKIIN or empty vector pcDNA3.1/myc-His (-) B was transfected into HL-60 cells by Lipofectamine 2000. Human CaMK II N proteins of transfected cells were detected by Western blot. Cell proliferation affected by human CaMKIIN was determined by MTT. Colony-forming assay was performed by soft agar growth system. The cells transfected with CaMKIIN were stained with Hoechst 33342 to detect the apoptotic proportion under fluorescence microscopy. Cell cycle was analyzed by flow cytometry.

RESULTSHuman CaMKIIN was stably transfected into HL-60 cells, and overexpression of human CaMKIIN inhibited the proliferation of HL-60/CaMKIIN cells compared to HL-60/mock cells and HL-60 cells [(0.44 ± 0.03) vs (0.94 ± 0.05) vs (0.94 ± 0.04), P<0.01]. The colony formation of HL-60/CaMKIIN was also markedly smaller[(21.00 ± 3.05)/500] than that of mock-transfected [(111.00±4.58)/500]] and control cells [(119.00±6.09)/500] (P<0.01). After 72 hrs-culture, the apoptotic proportion in cells transfected with CaMK II N was obviously higher than of cells transfected with mock DNA or control [(22.49 ± 2.15)% vs (7.17 ± 0.72)% vs (6.40 ± 0.55)%, P<0.01]. Up to (82.97 ± 2.90)% human CaMKIIN/HL-60 cells were arrested at G0/G1 phase, which was more than mock-transfected [(40.53 ± 2.38)%] and control cells [(41.63 ± 2.27)%] (P<0.05). Human CaMKIIN could down-regulate expression of Bcl-2 in transfected cells.

CONCLUSIONCaMK IIN up-regulation could inhibit proliferation and induce apoptosis of human acute myeloid leukemia cell HL-60.

Apoptosis ; Cell Proliferation ; Genetic Vectors ; HL-60 Cells ; Humans ; Proteins ; genetics ; metabolism ; Transfection ; Up-Regulation

OBJECTIVETo explore the main points of clinical differentiation between hemangioma and vascular malformation in infant.

METHODSBased on Mulliken and Waner's classification, from March, 1997 to February, 1999, 81 baby patients with hemangioma were included in this study. Thirty-eight cases, 43 cases received medical treatment of steroids.

RESULTSAll the patients were followed up from 5 to 7 years. Thirty-eight cases of red strawberry-like lesions limited in the skin began to involute within two years old. Of the 30 patients with strawberry-like lesions and subcutaneous mass, 20 cases involuted in varying degree; 10 cases' subcutaneous mass grew gradually and didn't involute, in 4 cases biopsy was performed, 3 cases were confirmed as hemangioma accompanied with venous malformation by pathology, 1 case was hemangioma accompanied with arteriovenous malformation. Of 13 cases with light blue or normal skin and subcutaneous mass, 7 cases involuted in varying degree; 6 cases grow gradually and didn't disappear, 2 cases were confirmed as venous malformation by biopsy.

CONCLUSIONSHemangioma in infant begins to involute within two years old. Vascular malformation or hemangioma with deep vascular malformation grows persistently and does not disappear. Skin temperature of lesion surface and dilative veins on the skin artery pulsation, are indexes compressibility, for differentiation between hemangioma and vascular malformation in clinical diagnosis.

Child, Preschool ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Hemangioma ; diagnosis ; Humans ; Infant ; Male ; Vascular Malformations ; diagnosis

OBJECTIVETo study the chemical constituents from the dried and nearly ripe fruits of Evodia (Euodia) rutaecarpa.

METHODThe compounds were separated and purified by solvent and chromatographic methods. Their structures were identified by spectroscopic techniques.

RESULTFifteen compounds were separated from the normal butanol extracts of the 70% aqueous ethanol extract of the dried and nearly ripe fruits of E. rutaecarpa. Among of them, four compounds were reported in the essay and identified as diosmetin-7-O-beta-D-glucopyranoside (1), isorhamnetin-3-O-rutinoside (2), diosmin (3) and chrysoeriol-7-O-rutinoside (4).

CONCLUSIONCompounds 1, 3 and 4 were separated from the dried and nearly ripe fruits of E. rutaecarpa for the first time.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Evodia ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Fruit ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure

In order to study the relationship between lengths of homologous fragments and chromsomic recombination rate in Saccharopolyspora erythraea, three homologous sequences, with mutant loci and different flanking sequences, (26bp + 27bp), (500bp + 576bp) and (1908bp + 1749bp), were synthesized by chemical reaction or PCR amplification, and cloned into pWHM3 to construct homologous recombination plasmids, pWHM1113, pWHM1116 and pWHM1119. When the plasmids were transformed into protoplast of Saccharopolyspora erythraea A226 under PEG mediated, on an average 30, 69 and 170 transformants grew on each plate for the three plasmids respectively, but chromosomic integration frequency were 0, 2% and 19% among corresponding transformants. Both pWHM1116 and pWHM1119 could take double crossover recombination, and exchange the mutant loci in the chromosome. It was concluded that when the flanking sequences were equal or more than (500bp + 576bp), they could take effective single and double recombination with Saccharopolyspora erythraea chromosome.
Chromosomes, Bacterial ; genetics ; Polymerase Chain Reaction ; Recombination, Genetic ; genetics ; Saccharopolyspora ; genetics

OBJECTIVETo evaluate the usefulness of serum alpha-fetoprotein (AFP) in clinical diagnosis and screening for hepatocellular carcinoma (HCC).

METHODSTotally 290 HCC patients, 48 liver cirrhosis patients, and 49 healthy subjects were enrolled in this study. Serum AFP analysis was performed to investigate the correlation between the serum AFP level in HCC and the clinical or biochemical parameters of the disease, which included the size and number of tumor and the TNM stage. Sensitivities and specificities of AFP in HCC prediction at different cut-off levels were determined.

RESULTSThe serum AFP level was significantly higher in HCC patients than in liver cirrhosis patients (P = 0.0274) and healthy subjects (P = 0.0001). Among 290 HCC patients, 95 patients (32.8%) were AFP-negative (AFP < 20 microg/L), 195 (67.2%) were AFP-positive (AFP > or =20 microg/L). Sensitivity and specificity of AFP at 20 microg/L cut-off was 67.2% and 29.2%, respectively, and the positive and negative predictive value was 85.2% and 12.8%, respectively. Sensitivity of AFP at 400 microg/L cut-off was only 42.8%. Serum AFP levels were significantly different among HCC with different tumor size (P = 0.0009), tumor number (P = 0.0001), and TNM stage [TNM I vs. TNM III-IV (P = 0.0001); TNM II vs. TNM III-IV (P = 0.0003)].

CONCLUSIONSIncreased serum AFP level is highly suggestive in HCC diagnosis. Combined with other imaging examinations, AFP level can be used for the screening of high risk population and for the follow-up of AFP-positive patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Diagnostic Techniques and Procedures ; Female ; Humans ; Liver Neoplasms ; blood ; diagnosis ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis