This study is to investigate the anti-allergic effect of anthocyanidin and to explore its possible mechanism. The experiments of passive cutaneous anaphylaxis reaction (PCA) and colorimetry were used to determine the effect of anthocyanidin on degranulation of mast cells in vivo. For in vitro study, various concentrations of anthocyanidin (100, 50 and 25 micromol x L(-1)) were added to the culture medium of mast cells cultured with 100 microg x L(-1) of dinitrophenyl (DNP) specific IgE overnight. The azelastine (100 micromol x L(-1)) was selected as the positive control. The antigen (DNP-human serum albumin, DNP-HAS)-induced release of degranulation was measured by enzymatic assay, histamine was determined by EIA, and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured by Western blotting, separately. In addition, the effects of anthocyanidin on phosphorylation of NF-kappaB, p38MAPK and Akt were observed by Western blotting. The results showed that treatments with anthocyanidin (100 and 50 mg x kg(-1)) were followed by a decrease in PCA of rats. Anthocyanidin (100 and 50 micromol x L(-1)) obviously suppressed the degranulation from mast cells, whereas results from anthocyanidin (100 and 50 micromol x L(-1)) group indicated significant inhibitory effect on histamine, the calcium uptake, TNF-alpha, IL-6, phosphorylation of NF-kappaB, p38MAPK and Akt of mast cells induced by antigen. Anthocyanidin may suppress the anaphylactic reaction by inhibiting the action of mast cells. NF-kappaB, p38MAPK and Akt at least in part contribute to this event.
Animals ; Anthocyanins ; pharmacology ; Anti-Allergic Agents ; pharmacology ; Calcium ; metabolism ; Cell Degranulation ; drug effects ; Histamine Release ; drug effects ; Immunoglobulin E ; immunology ; Interleukin-6 ; metabolism ; Male ; Mast Cells ; immunology ; metabolism ; physiology ; Passive Cutaneous Anaphylaxis ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective: To analyze and review the epidemiology , clinical features , treatment and prognosis of brucellosis ,in order to raise awareness of brucellosis in clinic .Methods:Eight clinical cases of brucellosis in Changzheng Hospital from 2010‐2015 were analyzed retrospectively ,and the clinical treatment experiences were concluded .Results:Five patients had history of epidemic exposure ,the epidemic exposure history of 3 cases were unknown .All patients had fever , fatigue and hyperhidrosis ,which 5 cases were accompanied by arthritis ,orchitis ,lymphadenectasis ,or migratory myalgia .The percentages of neutrophils in 8 cases were normal or relatively low ,and the erythrocyte sedimentation rate (ESR) was elevated in all patients ,in addition ,some indexes of liver function were elevated in 3 cases .All patients were cured by anti‐infective therapy , one patient who re‐contacted with goat was recrudescence .Conclusions: Brucellosis shows various clinical manifestations .Patients in non‐epidemic areas who have long‐term fever ,fatigue ,sweating or joint and muscle pain should be guarded against brucellosis ,and provide patients with early diagnosis and anti‐infective therapy .

This study was purposed to construct a vector containing human suppressor gene p53 and p16, and to investigate their expression and effect on K562 and HL-60 cells. pBudCE4.1-53-16 is a vector designed for simultaneous expression of human suppressor gene p53 and p16 in mammalian cell line. After transfection into K562 cells with lipofectamine(TM) 2000, the expression of p53 and p16 genes was detected by Western blot and immunocytochemical method. The growth curve, apoptosis, cell cycle were assayed by CCK-8 and flow cytometry. The results showed that the recombinant plasmid pBudCE4.1-53-16 was constructed successfully and were verified by PCR and restriction analysis. The expression of P53 and P16 protein could be detected after transfection into leukemia cells (K562 and HL-60) for 48 hours. As compared with control group, the cell proliferation in experimental group was inhibited, the cells were arrested in G0 phase and apoptotic cells increased (p<0.001). It is concluded that the recombinant plasmid pBudCE4.1-53-16 has been established. p16 and p53 in the recombinant plasmid pBudCE4.1-53-16 synchronously express in leukemic cells after transfection in vitro for 2 days and results in reduced proliferation, G0 arrest and apoptosis increase.
Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Gene Expression ; Genes, p53 ; Genetic Vectors ; HL-60 Cells ; Humans ; K562 Cells ; Plasmids ; Transfection

The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence (swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV. A rapid, sensitive and stable TaqMan Real-time RT-PCR assay was established, and its specificity and sensitivity were assessed, and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed. The results found that the crossing points showed linearly proportional to the logarithm of the input copy number. The correlation coefficient (R2) and the slope value of the standard curves with plasmid DNA were 0.994 and -3.312, respectively. The efficiency (E) of the PCR was 100%. Coefficients of variation values of the different diluted plasmid DNA were low in the same or different repeated experimental group. In addition, the assay was able to correctly detect genotype 4 HEV RNA from swine fecal samples. The sensitivity of established assay was 100-fold higher than that of conventional RT-PCR and 10-fold higher than nested RT-PCR.
Animals ; DNA Primers ; genetics ; Disease Reservoirs ; virology ; Feces ; virology ; Fluorescence ; Genotype ; Hepatitis E ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Swine ; virology

OBJECTIVETo explore efficacy of imatinib for patients with chronic myeloid leukemia(CML) and its resistance-related factors during the treatment.

METHODSThe clinical data of 214 CML patients received imatinib were analyzed respectively in our hospital from April 2005 to December 2010. The therapy history and efficacy of regular follow-up and factors influencing drug resistance were analyzed. COX regression analysis was used to perform the univariate and multivariate analysis.

RESULTSUntil the end of follow up, thirty-one patients (14.5%) occurred drug resistance. One of them was in accelerated phase(AP), and two in blast phase(BP); 69.2% of patients achieved a complete cytogenetic response(CCyR), and 31.3% of patients achieved a major molecular response(MMR). COX analysis was performed in 207 chronic phase(CP) patients. Univariate analysis showed that the course of disease before treatment, the hemoglobin count, the white blood cell count, whether achieved CCyR or not and whether achieved MMR or not were the influencing factors for imatinib resistance. Multivariate analysis showed that whether achieved CCyR or not was the independent factor for drug resistance.

CONCLUSIONWhether achieved CCyR or not is an independent factor and also a protective factor for imatinib resistance in patients with CML.

Adolescent ; Adult ; Aged ; Benzamides ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Neoplasm ; Female ; Follow-Up Studies ; Humans ; Imatinib Mesylate ; Leukemia, Myeloid, Chronic-Phase ; drug therapy ; Male ; Middle Aged ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use ; Retrospective Studies ; Young Adult

OBJECTIVETo report a case of T cell acute lymphoblastic leukemia (ALL) with t(1;19)(q23;pl3) and E2A-PBX1 fusion gene, which is a characteristic translocation of childhood B cell ALL (B-ALL).

METHODSThe chromosome, karyotype, immunophenotype and mRNA for fusion gene of the leukemic cells were examined by cytogenetic analysis, flow cytometry (FCM) and reverse transcriptase PCR (RT-PCR), respectively.

RESULTSThe cytogenetic karyotype of the patient was 47, XY, 9p+, 15p+, 17q-, der(19), t(1;19)(q23;pl3)\[5\]/46, XY\[15\], and E2A-PBX1 was positive. The leukemic cells expressed T cell markers. The patient was induced with hyper CVAD regimen (cyclophosphamide, vincristine, adriamycin, and dexamethasone), and achieved complete remission with normal cytogenetic karyotype 46 XY\[10\], and negative E2A-PBX1.

CONCLUSIONt(1;19)E2A-PBX1(+) can be implicated in adult T-ALL, besides childhood B-ALL.

Adult ; Chromosomes, Human, Pair 1 ; genetics ; Chromosomes, Human, Pair 19 ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Karyotyping ; Male ; Oncogene Proteins, Fusion ; genetics ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; Translocation, Genetic

Objective:The aim of this study was to research the relationship between HPLC fingerprint and anti-inflammatory effect of Zhideke granules， and the substance basis of its anti-inflammatory effect was preliminary explored. Method:The fingerprint of 10 batches of Zhideke granules were determined by HPLC， the mobile phase was consisted of methanol-0.2% phosphoric acid solution for gradient elution with a detection wavelength of 254 nm. Similarity analysis， hierarchical cluster analysis （HCA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to evaluate the quality difference between batches of Zhideke granules. The correlation analysis between the common peaks and the inhibition rates of Zhideke granules on ear swelling and cotton ball granuloma in mice was carried out by partial least squares （PLS）， and the peaks greatly contributing to the anti-inflammatory effect were screened out. Result:There were 31 common peaks in the HPLC fingerprint of Zhideke granules. The similarities of 10 batches samples were ≥0.992. The HCA and PCA analysis results were consistent， and the samples were divided into 3 categories. Combined with the OPLS-DA results， 15 components were the main markers affecting the differences of different batches of samples. Different batches of Zhideke granules differed in anti-inflammatory effect. The chromatographic peaks being positively correlated with the anti-inflammatory effect were mainly from Belamcandae Rhizoma and Scutellariae Radix， Chromatographic peaks 3， 6， 19， 27-30 had significant contribution to anti-inflammatory effect， of which peaks 28 and 30 were irisflorentin and wogonin. Conclusion:HPLC fingerprint combined with chemical pattern recognition method can provide a reference for systematic evaluation of the overall quality of Zhideke granules. Zhideke granules has a certain inhibitory effect on acute and chronic inflammation in mice， and the anti-inflammatory effect is the result of the combined action of various ingredients， while Belamcandae Rhizoma and Scutellariae Radix have significant significance for the anti-inflammatory effect.