Objective Establish rabbit VX2 soft tissue tumor model,and treat it with percutaneous ethanol injection(PEI)under the guidance of magnetic resonance imaging.Make ready for the therapeutic evaluation with functional magnetic resonance imaging. Methods Fifteen healthy New Zealand white rabbits were included in this study.0.2 mL tumor tissue suspensions were injected into the rabbits’posterior limb.14 days later,all rabbits were underwent conventional MRI examination.PET were performed to all the tumors under the guidance of MRI in the next day of the examination.T2 WI was used as guidance and monitoring means.MR com-patible puncture needle with lateral hole was stabed into the lesion center,and inj ected anhydrous ethanol according to the volume of tumors’diameter (1 mL/cm )slowly.the tumors signal characteristics,morphological feature and pathological feature were ob-served pre and post-operation.Results All of the 1 5 rabbits were established soft tissue tumor model successfully;the success rate is 100%.The tumors were oval or round,3-4 cm in diam.MRI scanning showed low signal on T1 WI and high signal on T2 WI be-fore PEI.PEI was performed to all the tumors under the guidance of MR successfully with 3.5 mL ethanol injected into the tumors in average.T2 WI could monitor the ethanol in dispersion and distribution within the tumors clearly.Histologically,tumors were composed of large,uniform,oval/round cells arranged in solid nests which was intensive in the periphery of tumors.Necrosis tissue was apparent in the center of the tumors.10 days after operation,most tissue in the periphery of tumors was coagulative necrosis , only a few tumor cells left.Ranges of necrosis in the tumors center were obviously increased compared with pre-operation.Conclusion Rabbit VX2 tumor of soft tissue model is suitable for the therapeutic evaluation of tumor .It is an animal model which has the characteristic of simple to operate and high rate of suc-cessful.MR T2 WI can monitor the ethanol in dispersion and distribution within the tumors clearly.It is a good guidance and monitoring imaging method of tumor ablation.

Toad venom is the Bufo bufo gargarizans or B. melanostictus after the ears of the gland secretion, used in the treatment of various cancers in recent years. Research shows that the main anti-tumor components in bufadienolide. Bufadienolide have free type structure and conjunct type structure. To identify and clarify the difference between bufogenin and bufotoxin contained in Bufonis Venenum, which was from B. bufo gargarizans, an UPLC-TQ-MS method has been established. UPLC-TQ-MS method was used to identify and quantify the major bufadienolides in Bufonis Venenum. UPLC-TQ-MS assay with positive ion mode was performed on a Waters ACQUITY UPLC BEH C, (2.1 mm x 100 mm, 1.7 µm) with the mobile phase consisting of 0. 1% aqueous formic and acidacetonitrile in gradient elution at a flow rate of 0.4 mL · min⁻¹ and the column temperature was set at 35 °C. By comparing their retention time and high resolution mass data of Bufonis Venenum extracts, 37 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were free-type bufadienolides (bufogenin), 11 of them were conjugated bufadienolides. There were significant differences in the main composition between fresh and processed Bufonis Venenum. The study found that the chemical composition of toad venom through great changes after processing, conjunct type content is much less, free type content as well change.
Amphibian Venoms ; chemistry ; metabolism ; Animals ; Bufonidae ; classification ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Molecular Structure ; Tandem Mass Spectrometry ; methods

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult

Objective To study the effects of Jianpi Xiaoai Prescription on cell cycle and apoptosis of human colon cancer HCT116 cells and related factors of Wnt/β-catenin signaling pathway; To investigate its mechanisms of anti-metastasis and recurrence of colorectal cancer. Methods The logarithmic growth phase HCT116 cells were divided into blank group, saline group, 5-Fu group, and Jianpi Xiaoai Prescription low-, medium-, and high-dose groups. After intervention for 48 h, the cells were harvested, and the cell cycle and apoptosis were detected by flow cytometry. The protein expression of β-catenin in the nucleus was detected by Western blot, and the expression of c-myc and cyclinD1 mRNA was detected by PCR. Results Compared with the blank group and saline group, the ratio of HCT116 cells apoptosis of Jianpi Xiaoai Prescription low-, medium-, and high-dose groups increased; the proportion of cells in phase G1 increased and the proportion of S cells decreased; the expression of β-catenin protein in the nucleus and the expression of c-myc,cyclinD1 mRNA decreased,especially in the Jianpi Xiaoai Prescription high-dose group,with statistical significance(P<0.05,P<0.01).Conclusion Jianpi Xiaoai Prescription can promote apoptosis of human colon cancer HCT116 cells and block the cell cycle, and its mechanism may be related to regulating Wnt/β-catenin signaling pathway.

Objective To analyze the clinical characteristics of concomitant vertigo in patients with sudden deafness (SD). Methods Ninety-six cases of SD were reviewed retrospectively from January 2005to July 2009. SD and benign paroxysmal positional vertigo (BPPV) were diagnosed according to the guides of China Medical Association. The characteristics of vestibular function and the order of the onset of cochlear and vestibular symptoms were analyzed. Results Of all 96 cases, 23 (24.0%) cases presented with BPPV;58(60.4% ) cases took the form of unilateral vestibular hypofunction and 15( 15.6% ) cases had normal vestibular function. Time interval between cochlear and vestibular symptoms was as follows: 46 patients could tell the exact time of onset of cochlear and vestibular symptoms, of which 6( 13.0% ) cases occurred simultaneously; 4 (8. 7% ) cases presented vertigo within 1 hour after onset of cochlear symptomar hypofunction; 21 (45.7%)cases showed time interval between 1 hour and 24 hours; and 13 (28.3%) cases presented vertigo at several days( less than 10 days) after cochlear symptoms. And only in 2 (4.3%) cases did vertigo occur before cochlear symptoms. Conclusions Concomitant vertigo in idiopathic SD took the forms of normal or abnormal vestibular function, some of which were BPPV. Occurrence of vertigo was after cochlear symptoms.

OBJECTIVETo research the maturation regulation of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) cell soluble antigens.

METHODSBCG HSP 70 was purified by SDS-PAGE electrophoresis and its biological activity was determined with ELISA. Phenotypes of DCs pulsed with antigens or with both antigens and BCG HSP 70 were analysed with flow cytometry. MTT assay was used to estimate the proliferation of self lymphocytes and the mixed lymphocyte reaction (MLR) of BCG HSP 70 primed DCs.

RESULTSThe characteristics of DCs had changed after loaded with soluble antigens of HCC. There were about 10% DCs which had lost their specific markers. The expression levels of CD54, CD83, CD86 molecules and the stimulatory ability in allogeneic MLR decreased. However, after being activated by BCG HSP 70, the DCs pulsed with antigens could keep their special markers and the expression levels of CD54, CD83, CD86 molecules increased too. The stimulatory abilities in allogeneic MLR and proliferation of self lymphocytes also improved.

CONCLUSIONThis study shows that BCG HSP 70 can induce DCs pulsed with antigens maturation and improve their antigen-presenting ability, which may be a useful maturation inducer for dendritic cells.

Antigen Presentation ; Antigens, CD ; analysis ; Antigens, Neoplasm ; immunology ; B7-2 Antigen ; Carcinoma, Hepatocellular ; immunology ; Dendritic Cells ; cytology ; immunology ; HSP70 Heat-Shock Proteins ; immunology ; Humans ; Immunoglobulins ; analysis ; Intercellular Adhesion Molecule-1 ; analysis ; Liver Neoplasms ; immunology ; Membrane Glycoproteins ; analysis ; Mycobacterium bovis ; immunology

AIM:To investigate the effect of ozone bath on the pathological changes and the expression of cyto-kines,platelet-derived growth factor (PDGF), transforming growth factor-β3(TGF-β3), and tumor necrosis factor-α (TNF-α),in the wounds of deep second-degree burns in rats. METHODS:Male clean-grade SD rats(n=80) were ran-domly divided into 2 groups, ozone bath group and routine dressing group (control group), with 40 rats in each group. Deep second-degree burn wound was established on the back of the rats,and then the examinations were conducted at 3 d, 7 d,14 d and 21 d after burn. For the routine dressing group,the wound was cleaned by normal saline and covered with io-dophor vaseline gauze every 2 d. For the ozone bath group,before the dressing,the rats were put into the clean foam box to accept ozone fumigation for 20 min(50 mg/L),and then accepted dressing change as the same as that in control group every 2 d. At each time point,the tissue specimens from these rat wounds(at wound center) were taken. The rats in ozone bath group received cleaning by saline cotton and then the ozone bath fumigation, while the rats in control group only re-ceived cleaning by saline. After that,the tissue specimens were taken again for HE staining,immunohistochemical staining and semiquantitative observation combined with image data analysis. The concentrations of the cytokines PDGF, TGF-β3 and TNF-α in the wound were measured by double-antibody sandwich ELISA. RESULTS:In ozone bath group, the wounds were smooth with clear edge and slight inflammatory reaction,swelling and exudation were weaker,and the wound healing rate was higher than that in control group with significant difference. Under microscopic observation with HE stai-ning,slighter inflammatory reaction in ozone bath group was observed than that in control group at each time point,and the numbers of fresh capillaries,fibroblasts and epithelial cells were significantly larger than those in control group. The ex-pression levels of PDGF and TGF-β3in the wound tissue homogenate in ozone bath group were higher,and the expression level of TNF-α was significantly lower than those in control group at each time point with significant difference. CONCLU-SION:The ozone bath therapy improves the local pathological changes and promotes the expression of cytokines PDGF and TGF-β3,which are associated with wound healing,as well as reduces the expression of inflammatory mediator TNF-α in the rats with deep second-degree burns,thus promoting the wound healing and anti-inflammatory responses.

OBJECTIVETo study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.

METHODSMethylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.

RESULTSHypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).

CONCLUSIONFHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.

Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Base Sequence ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Myelodysplastic Syndromes ; classification ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics

OBJECTIVETo evaluate anti-HEV diagnostic kits by experimental infecting rhesus monkeys with HEV.

METHODSEight rhesus monkeys were infected with genotype 1 and 4 HEV separately. The alanine aminotransferase (ALT) level of all monkeys were detected before and after the process of infection. HEV RNA in stool specimens was tested by reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Anti-HEV IgG in serum was detected by GL-IgG and WT-IgG.

RESULTSHEV RNA presented in the stool of all the 8 monkeys after infection. The ALT level of 1 monkey infected with genotype 1 HEV and 2 monkeys infected with genotype 4 HEV appeared abnormally after infection. Tested by GL-IgG, 2 of the 4 monkeys infected with genotype 1 HEV and 1 of 4 monkeys infected with genotype 4 HEV seroconverted to anti-HEV IgG. However, when tested by WT-IgG, all the infected monkeys seroconverted to anti-HEV IgG. The anti-HEV IgG tested by WT-IgG was positive during the whole observation period,and the anti-HEV IgG measured by GL-IgG only remained 12 weeks after infection. Detected by GL-IgG and WT-IgG, seropositive conversion of the anti-HEV IgG happened almost at the same time.

CONCLUSIONBoth GL-IgG and WT-IgG could detect the anti-HEV IgG of experimentally infected rhesus monkeys but the WT-IgG had a higher sensitivity for detection of anti-HEV IgG than

Alanine Transaminase ; blood ; Animals ; Disease Models, Animal ; Genotype ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; Immunoglobulin G ; immunology ; Macaca mulatta ; Reverse Transcriptase Polymerase Chain Reaction