Objective To describe the sonographic appearance of mesenchymal hamartoma of the liver(MHL)and to analyze the diagnostic value of ultrasound.Methods Eleven surgically and pathologically confirmed cases of MHL from January 2005 to May 2011 in the Beijing Children′s Hospital were retrospectively reviewed.Results Ultrasound examinations showed 9 cystic hamartomas,including 7 multiseptate cystic and 2 monocystic lesions.Of the 7 multiseptate cystic hamartomas,4 had a honeycomb appearance and 3 had irregularly-distributed multiple cysts with varied septations.Of the 2 monocystic hamartomas,1 had a large cystic portion while the other was mainly solid with approximately 4% cystic portion of the tumor.Two cases in this group were solid,presenting with a well-defined homogenous mass.Conclusions Ultrasonography is an effective imaging modality for the diagnosis for MHL.A mixed or a mainly-cystic liver mass found in a child less than 2 years old should be suspicious for MHL.

Objective To explore the interaction between serum melatonin and the status of disease and probe the effect factor of serum melatonin change in asthmatic children.Methods Serum melatonin was measured in asthmatic children with 15 cases of mild persistent asthma,15 cases of moderate persistent asthma,15 cases of severe persistent asthma,15 cases of stable asthma and 15 cases of normal subjects by enzyme-linked immunosorbent assay(ELISA).Results The levels of serum melatonin in the 5 groups of mild persistent asthma,moderate presistent asthma,Severe Persistent asthma,Stable asthma,control subject were(22.76?5.16)ng/L,(16.79?3.35)ng/L,(11.54?1.45)ng/L,(22.06?3.36)ng/L,(28.72?4.32)ng/L,respectively.There were significant differences between any of them(Pa

OBJECTIVE: To investigate the culture of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases (CHD) before and after percutaneous coronary intervention (PCI), and to observe the cells shape and determine the cell number and proliferation activity. METHODS: Ninety-five patients were divided into a CHD group(n=65) and a control group (n=30). The mononuclear cells were isolated from peripheral blood of patients with CHD before, right after and 4 days after PCI by Ficoll-density centrifugation. The isolated cells were cultured in RPMI1640 medium supplemented with VEGF165 and bFGF.EPCs were characterized as adherent cells of double positive for DiL-acLDL uptake and FITC-UEA-I binding by direct fluorescent staining under a fluorescence microscope. The EPCs specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter analysis. The cell shapes were analysed and the number of colony-forming units(CFU) was counted by phase-contrast microscope. RESULTS: The number of EPCs reduced in patients with CHD before the PCI, but the cell number was significantly increased in patients with CHD after the PCI, and the number reduced in patients with CHD 4 days after the PCI. How-ever, the number of CFUs did not change in patients before and after the PCI. CONCLUSION PCI can increase endothelial progenitor cells in patients after the PCI; but 4 days after the PCI, this increase will not exist.
Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Cell Adhesion ; Cell Count ; Cell Movement ; Cells, Cultured ; Coronary Disease ; blood ; therapy ; Endothelial Cells ; pathology ; Female ; Humans ; Male ; Middle Aged ; Stem Cells ; pathology

OBJECTIVETo develop a recombinant vaccinia virus vaccine expressing HPV58 E7 and to determine its immuno-protective activity in mice bearing HPV58 E7+ tumor.

METHODSE7 DNA was amplified and cloned from a plasmid containing HPV58 E7 genome by PCR. To abolish its transforming activity, the nucleotides coding for amino acid residues at positions 24 and 92 were modified by site-directed mutagenesis so that cysteine was substituted by glycine. Balb/c 3T3 cells were transfected with mE7. The expression of E7 protein by the mE7-transfected Balb/c cells was confirmed by immunofluorescence staining. The transfected cells were observed in vitro for anchorage-independent growth and tumorigenesis in nude mice. Recombinant E7 vaccinia virus vaccine was constructed by homologous recombination of HPV58 E7 vaccinia expression plasmid and vaccinia virus (Tiantan stain). The immuno-protective activity of the vaccines was determined by tumor growth inhibition and cytotoxic T lymphocytes (CTL) induction in vaccine-immunized syngeneic mice.

RESULTSSubstitution of cysteine by glycine at both positions 24 and 92 of HPV58 E7 abolished its transforming activity. Growth of HPV E7+ tumor in mice immunized with the recombinant vaccinia virus expressing HPV58 E7 was inhibited, and the surviving time of the immunized mice was prolonged. CTL activity was induced as revealed by in vitro cytotoxicity assay using E7+ tumor cells as target cells.

CONCLUSIONSHPV58 E7, with its transforming potential abolished, may be used as vaccine for immunotherapy of patients with HPV 58 related cancers.

Animals ; Mice ; Mice, Inbred BALB C ; Mutagenesis, Site-Directed ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, Synthetic ; immunology ; Vaccinia virus ; genetics ; immunology

Objective To analyze clinical diagnosis and management of 5 patients with actinomycete keratitis.Design Retro- spective case series.Participants 5 patients (5 eyes) with actinomycete keratitis.Methods The clinical features and microbiologic da- ta of 5 culture-proven cases of actinomycete keratitis recorded between October 2004 to March 2006 were analyzed.Main Outcome Measures clinical characteristics,isolations identification,drug susceptibility test and treatments.Results All patients were males and farmers.Of the 5 cases presented in this study,4 cases were followed by minor trauma as a predominant risk factor,and were pre- sented by a chronic progressive corneal ulcer with a wreath pattern of infiltrate.The diagnosis of all cases was based on laboratory in- vestigations,by which 4 cases of nocardia and one case of streptomyce were identified.A variable drug sensitivities were presented in nocardia isolates,which including TMP-SMZ,amicasin,gentamicin and fluorine-quinolones.Conclusions Nocardia keratitis is mainly followed by a minor trauma.It is identified predominantly by laboratory investigations.Tropical and systemically sensitive biotic are the initial choice,while debridement and amnionic transplantation could be an effective alternative.

Human placental decidua basalis-mesenchymal stem cells (PDB-MSCs) are multipotent cells from the human term placenta, which are ethically conducive, easily accessible and high-yielding source. PDB-MSCs can differentiate into adipogenic, osteogenic and neurogenic cells under appropriate conditions, which may be an attractive and alternative source of seed cells for tissue engineering. To investigate the effect of hypoxia (1% O2) on human PDB-MSCs and the expression of cytokine, PDB-MSCs were isolated from human placenta by density gradient centrifugation and cultured in the Dulbecco's modified Eagle's medium-high glucose (DMEM-HG) containing 10% fetal bovine serum (FBS), and the fifth passage of PDB-MSCs were taken. PDB-MSCs were divided into 4 groups according to the concentrations of O2 and FBS: 20% O2, 10% FBS; 20% O2, 0% FBS; 1% O2, 10% FBS; 1% O2, 0% FBS. The proliferation and apoptosis of PDB-MSCs were detected by MTT and flow cytometric analysis at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, and 96 h, respectively. Vascular endothelial growth factor (VEGF) released from PDB-MSCs was detected by enzyme-linked immunosorbent assay (ELISA) at the same time points. The results showed that hypoxia enhanced the proliferation of PDB-MSCs at 12 h under the condition of 10% FBS, while at 24 h under the condition of 0% FBS (P<0.01, n=3). In normoxia, the cells cultured in 10% FBS displayed a significant proliferation compared to those cultured in 0% FBS. However, in hypoxia, the number of cells cultured in 0% FBS (serum deprivation) increased significantly compared to that cultured in 10% FBS at 24 h and 96 h respectively (P<0.05, P<0.01, n=3). With the flow cytometric analysis of cell apoptosis under the condition of hypoxia and serum deprivation, we found that hypoxia and serum deprivation did not induce PDB-MSCs apoptosis (P>0.05, n=3). This conclusion may relate to the expression of VEGF which needs further research. In conclusion, the results obtained indicate that PDB-MSCs are able to bear hypoxia and serum deprivation, suggesting that PDB-MSCs can be used as seed cells for ischemia related tissue engineering.
Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Decidua ; cytology ; Female ; Humans ; Mesenchymal Stromal Cells ; cytology ; Placenta ; cytology ; Pregnancy ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; metabolism

A pairs of primers were designed according to the fusion protein(F)gene sequences of canine distemper virus(CDV)in GenBank.A 369 bp fragment aimed signal peptide fragment of F gene was amplified.The PCR products from viscera samples,blood,urine of fur animals including foxes,minks and raccoon dogs,which collected in the years 2005-2007,were cloned to pMD18-T Vector and sequenced.We obtained 13 positive signal peptide fragments from wild-type strains.The results indicated there was obviously genetic diversity between the wild-type strains and CDV3 and other vaccine strains.The homology with CDV3 is 80.7%-83.2%in nucleotide,and 64.8%-71.3%in amino acid.The analysis for the hydrophobic regions indicated the function of signal peptide fragment may be changed.This study can offer aca- demic data to research of CDV genetic variation and epidemiology.

OBJECTIVETo characterize genotypic resistance within HBV RT region in chronic hepatitis B (CHB) patients with nucleos(t)ide analogue (NA) treatment.

METHODSSerum samples of 229 CHB patients with NA treatment were obtained. Full-length HBV RT sequences were amplified, sequenced and analyzed, on the following NA resistant (NAr) mutations belonging to different NAr pathways.

RESULTSAmong 229 HBV isolates, 14.41% (33/229) and 85.59% (196/229) were genotype B and C, respectively; and the patients with HBV genotype C may be more susceptible to develope resistant mutations than patients with HBV genotype B(chi2 = 2.95, P < 0.05). NAr mutations were detected in 63 CHB patients. Mutations were not found at rtI169, rtT184, rtA194 or rtS202. RtM204 mutations were detected at the highest frequency among 63 mutants (40/63, 63.49%) and found to display 11 combination mutation patterns, in which rtM204I were associated with rtL80I/V and rtL180M, and rtM204V were associated with rtL1l80M, respectively. Conclusions There are complicated mutation patterns in the HBV RT region for chronic hepatitis B (CHB) patients with nucleos(t)ide analogue (NA) treatment. RtM204V/I mutation was the highest.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Male ; Middle Aged ; Mutation ; drug effects ; Nucleosides ; therapeutic use ; Nucleotides ; therapeutic use ; RNA-Directed DNA Polymerase ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism ; Young Adult

OBJECTIVETo explore the killing effect of the mutant herpes simplex virus thymidine kinase (HSV-sr39tk) and its wild-type (HSV-tk) mediated by lentiviral vector on T lymphocytes in vitro and compare T cell survival rate after GCV or ACV treatment.

METHODSThe three-plasmid lentiviral vector system including packaging plasmid DeltaNRF, envelope plasmid VSV-G and vector plasmid (pTK151 + HSV-sr39tk or pTK151 + HSV-tk) were cotransfected into human embryonic kidney 293T cells using modified calcium phosphate precipitation methods. The packaged virus was harvested 72 h later. The survival of T cells expressing HSV-sr39tk or HSV-tk was measured by MTT assay after 4 day-culture against a gradient of GCV or ACV concentrations.

RESULTSThe three plasmids were effectively cotransfected and a high titre of lentivirus was obtained (2 x 10(6) IU/ml). 39tk(+) T cell survival rates declined promptly when the prodrug GCV/ACV concentrations increased from 0 micromol/L to 10 micromol/L. The T cell survival rates in GCV group declined from (96.04 +/- 3.23)% to (36.76 +/- 4.38)% while in ACV group from (97.31 +/- 4.61)% to (43.75 +/- 8.99)%. However, when GCV/ACV concentrations were more than 10 micromol/L, further decline of 39tk(+) T cell survival rates became unobvious. The growth rate of 39tk(+) T cell exposed to GCV or ACV was obviously lower than that in un-transfected T cells (P < 0.05). Tk(+) T cells were sensitive to GCV (P < 0.05), but not to ACV (P > 0.05). There was a significant difference in killing effects between 39tk(+) T cell + GCV group and tk(+) T cell + GCV group (P < 0.05).

CONCLUSIONThe lentiviral vectors containing HSV-sr39tk gene could infect T lymphocytes effectively and stably without affecting the proliferation of the transduced cell. In contrast to HSV-tk gene, T cells infected HSV-sr39tk were more sensitive not only to GCV but also to ACV.

Acyclovir ; pharmacology ; Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Ganciclovir ; pharmacology ; Genetic Vectors ; Lentivirus ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; T-Lymphocytes ; cytology ; drug effects ; Thymidine Kinase ; genetics ; Transfection

BACKGROUNDAscorbic acid (AA) represents one of the most important enzyme co-factors, antioxidants and neuromodulators and plays an important role in the cerebral system. Increasing evidence has suggested that AA could treat certain kinds of vertigo diseases such as Meniere's disease. To elucidate the neurochemical functions associated with AA in vertigo, the change of extracellular AA in the brain cortex following caloric vestibular stimulation (CVS) was evaluated.

METHODSAn on-line electrochemical detection was coupled with in vivo microdialysis to continuously monitor the change of extracellular AA in the primary somatosensory (SI) area of guinea pigs following a caloric vestibular stimulation. Sixteen guinea pigs were divided into three groups, i.e., experimental group with irrigation of the ear canal with ice water (0 degrees C) (n = 8), and two control groups, one with irrigation of the ear canal with warm water (38 degrees C) (n = 4) and the other with irrigation of the auricle with ice water (n = 4).

RESULTSIn the experimental group, the ice water irrigation of the left external ear canal induced a horizontal nystagmus towards the right side lasting about 45 seconds. No nystagmus was induced by warm water irrigation of the external ear canal or by ice water irrigation of the auricle. The extracellular AA concentration significantly increased following the ice water vestibular stimulation, reaching a maximum of (130 +/- 20)% (n = 8) of the basal dialysate level (2.61 +/- 0.92) micromol/L (n = 8), lasting at least for an hour. AA level did not change distinctly after the irrigation of the left external ear canal with warm water or the irrigation of the auricle with ice water.

CONCLUSIONSThe concentration of extracellular AA in the brain cortex of the SI area increased following the ice water vestibular stimulation. This demonstration may be useful for the investigation of the neurochemical processes associated with AA in the process of vertigo.

Animals ; Ascorbic Acid ; analysis ; Cerebral Cortex ; metabolism ; Electrochemistry ; methods ; Extracellular Space ; metabolism ; Guinea Pigs ; Ice ; Male ; Microdialysis ; methods ; Physical Stimulation ; methods ; Vestibule, Labyrinth ; physiopathology