Objective To research the expression,methylated regulation and clinical significances of miR-34b in chidren with acute leukemia(AL).Methods The methylation status of miR-34b promoter CpG islands were detected with methylation-specific polymerase chain reaction (MSP) in patients with AL.Then the expression of miR-34b was compared,which was detected by Taqman real-time fluorescence quantitative polymerase chain reaction (qRT-PCR),between the AL patients and normal group,in order to analyze their relationship with the clinical indicators.Resuits In 8 leukemia cell lines (U937,HL-60,MV4-11,M2R,K562,Raji,CCRF,DAMI) showed methylation,the positive rate of the methyl was 100％.Thirty-one acute lymphocytic leukemia(ALL) pediatric patients were newly diagnosed,and 24 cases showed methylation,the methylation positive rate 77.42％ (24/31).Nineteen acute myeloid leukemia(AML) patients were newly diagnosed,and 8 cases showed methylation,the methylation positive rate was 42.11％ (8/19 cases).There was no methylation in the 23 cases of normal children.The relative expression levels of miR-34b in the normal group,the group of leukemia cell lines,the group of ALL pediatric patients with newly diagnosed,the AML group and the group with mixed lineage leukemia gene rearrangement MLL+ were 5.22 ± 1.15,0.03 ± 0.03,1.65 ± 0.69,0.18 ± 0.06,0.64 ± 0.34,respectively.The findings indicated that there were significant differences in the relative expression levels of miR-34b between the normal group and the group of leukemia cell lines,the ALL group,the AML group,and the MLL+ group.The relative expression and methylated level of miR-34b had no statistically difference in gender,age at diagnosis,WBC count,chromosome,fusion gene,MLL gene rearrangement,and the minimal residual disease(LDH) levels in newly diagnosed AML patients(all P ＞ 0.05).And the relative expression and methylated level of miR-34b had no statistically difference in gender,age at diagnosis,WBC count,immunophenotype,chromosome,fusion gene,MLL gene rearrangement,TEL/AML1 gene,risk stratification,the minimal residual disease (MRD) in thirty-three days and the LDH levels in newly diagnosed ALL patients(all P ＞ 0.05).But as for the response to prednisone experiment,there was a significant difference between the sensitive group and the non sensitive group(P ＜ 0.05).Conclusions The expression level of miR-34b in AL was significantly lower and it was regulated by methylation mechanism,which implies that miR-34b may play a role of a tumor suppressor gene in the pathogenesis of leukemia.MiR-34b may affect the early treatment response of ALL patients,and it may be an indicator of risk stratification and poor prognosis in pediatric ALL.

Processed Chinese medicine is the core of traditional Chinese medicine (TCM) industry chain,which directly affects the clinical efficacy and. safety of Chinese patent medicine and clinical formula Decoction pieces. Studied the variation of effective substance in vivo Chinese medicine processing before and after processed, clarifying the effective substance and processing principle is a top priority of the development of Chinese medicine processing. The traditional research method chiefly focus on the variation about chemicals in vitro of processed Chinese medicine, it cannot reveal that the integrity and complexity of processed Chinese medicine efficacy changes, so the change process is the focus of future research in vivo on the base of effective substance of TCM This paper described the research on the base of effective substance of TCM and Processed Chinese medicine research status in vitro, discussed the analytical methods (plasma chemistry, pharmacokinetics, metabonomics) of the dynamic process in vivo about processed Chinese medicine, and pointed out development and related problems in process in vivo on the base of effective substance of TCM, which could provided research ideas and methods for in-depth interpretation of Chinese medicine processing mechanism.
Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Humans ; Medicine, Chinese Traditional

Gardeniae Fructus contains volatile ingredients, however, the species and proportions in different processed products of Gardeniae Fructus are different. In this experiment, volatile ingredients were separated by steam distillation with content of 1.2, 1.0, 0.9, 0.7 µL · g(-1) in Gardeniae Fructus, fried Gardeniae Fructus, stir-baked Gardeniae Fructus, Gardeniae Fructus fried into carbon respectively. One hundred and twenty-four kinds of volatile components were identified by GC-MS. Fifty-three kinds of volatile ingredients consisted in Gardeniae Fructus accounting for 93.85%, 54 kinds in fried Cardeniae Fructus accounting for 92.01%, 32 kinds in stir-baked Cardeniae Fructus accounting for 91.59% and 43 kinds in Gardeniae Fructus fried into carbon accounting for 90.81%. In this paper, analysis of Gardeniae Fructus by GC-MS provides a scientific basis for elucidating the mechanism of different processed products.
Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; Gardenia ; chemistry ; Gas Chromatography-Mass Spectrometry ; Molecular Structure ; Volatile Organic Compounds ; chemistry

OBJECTIVETo evaluate the values of whole body diffusion weighted imaging (DWI) in screening primary unknown tumor in patients with metastases.

METHODSTotally, 34 patients with metastases of primary unknown tumors were scanned with whole body DWI, and conventional magnetic resonance (MR) imaging was performed if suspected lesions were detected. All the metastases including 27 cases of osseous metastases, 2 brain metastases, 2 liver metastases, 1 pulmonary multiple metastasis, 1 neck metastasis and 1 malignant ascites, were diagnosed by computed tomography, single photon emission computed tomography, or MR imaging. For the proven primary tumors diagnosed by biopsy or pathology of surgical specimens, apparent diffusion coefficient (ADC) values of the primary and metastatic lesions were measured respectively. The sensitivity and specificity of this technique for screening primary tumors were evaluated.

RESULTSWe found 24 cases with suspected primary lesions, in which 23 lesions were proved to be primary tumors, and 1 was proved to be benign lesion. And no definite primary lesion was found in 10 cases on whole body DWI, but in which 1 case was diagnosed with primary tumor by biopsy later, and the other 9 cases remained unknown within follow-up of over half a year. The difference was not significant in ADC values between primary and metastatic lesions (P>0.05). The sensitivity and specificity of whole body DWI for searching primary tumors was 95.8% and 90.0%, respectively.

CONCLUSIONCombined with conventional MR scanning, whole body DWI can help to search primary lesions of patients with metastases.

Adult ; Aged ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; diagnosis ; pathology ; Neoplasms ; diagnosis ; pathology ; Sensitivity and Specificity ; Whole Body Imaging ; methods

OBJECTIVETo study the expression level and CpG island methylation status of miR-34b in leukemia cell lines and to research the effect of DNA methylation on the proliferation of leukemia cells regulated by miR-34b.

METHODTaqman real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was carried out to detect the relative expression of miR-34b in control group (bone marrow cells of 20 children without blood disease) and 8 leukemia cell lines (U937, HL-60, MV4-11, M2R, K562, Raji, CCRF, DAMI). Methylation-specific polymerase chain reaction (MSP) was carried out to detect the methylation differences of miR-34b in control group (Bone marrow cells of 23 children without blood disease), 8 leukemia cell lines. HL-60 and K562 were treated with methyltransferase inhibitor (5-aza-2-dC) for further detection of its methylation status and expression of miR-34b. Hsa-miR-34b mimics was transfected into K562 cell by liposome, the transfection efficiency was detected by flow cytometry. The cell proliferation of hsa-miR-34b transfected group in each stage was measured with CCK-8 assay, and then compared with non-transfected group and negative control group.

RESULTThe relative expression level of miR-34b in the group of children without blood disease and the group of leukemia cell lines were 5.22 ± 1.15, 0.03 ± 0.03. The results showed that, the group of leukemia cell lines was significantly different from the control group (t = 4.538, P < 0.01) . Eight leukemia cell lines showed methylation, the positive rate of the methyl was 100%. There was no methylation in the 23 cases of control group. After leukemia cell lines HL-60 and K562 were treated with 5-aza-2-dC, the methylated bands became obviously weakened, and the relative expression levels of miR-34b substantially increased 49.5 times and 18.8 times respectively. After hsa-miR-34b mimics was transfected into K562 cell by liposome, its transfection efficiency detected by flow cytometry was 61% and the cell proliferation was measured with CCK-8 assay from which it was found that the cell proliferation was significantly suppressed compared with the control group at 48 h (t = 9.303, P < 0.01), 72 h (t = 65.617, P < 0.01), 96 h (t = 36.878, P < 0.01) and 120 h (t = 18.748, P < 0.01) in hsa-miR-34b transfected group, with the inhibition rate of 12.2% (48 h), 45.7% (72 h), 32.5% (96 h) and 22.9% (120 h).

CONCLUSIONThe hypermethylation of promoter leads to decrease in the expression levels of miR-34b in leukemia cell lines, which attenuate mechanism of proliferative inhibition may be one of the reasons of occurrence or development of childhood leukemia.

Azacitidine ; Cell Line, Tumor ; Cell Proliferation ; genetics ; CpG Islands ; DNA Methylation ; HL-60 Cells ; Humans ; Leukemia ; genetics ; MicroRNAs ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction ; Transfection

AIMTo study the effects of ginkgolides (Gin) on the expression of hypoxia-inducible factor-1alpha (H1F-1alpha) in primary cultured cortical neurons treated with CoCl2 and the relationship with ERK signal pathway.

METHODSWe observed the effects of Gin (37.5 mg/L) on morphology and viability on primary cultured cortical neurons with treatment of CoCl2 (125 micromol/L). The expression of HIF-1alpha and p-ERK of neurons induced by CoCl2 pretreated with Gin were assessed by Western-blot. We analyzed the relationship between HIF-1alpha expression activated by Gin and ERK signal pathway with treatment of PD98059 (100 micromol/L), a selective inhibitor of ERK.

RESULTSIt was shown that Gin had protective effects on CoCl2 damaged neurons by raising the neuronal viability. Some basic expression of HIF-1alpha and p-ERK were observed in normal cultured cortical neurons. The expression of HIF-1alpha and p-ERK increased strikingly when treated with CoCl2 for 4 h. The levels of HIF-1alpha and p-ERK increased even more in the neurons pretreated with Gin for 24 h before CoCl2. The levels of HIF-1alpha and p-ERK were notably inhibited with pretreatment of PD98059, while Gin could prevent this inhibition.

CONCLUSIONGin has protective effects on neurons damaged by CoCl2 which might be related to the increase of the level of HIF-1alpha and the activation of ERK signal pathway.

Animals ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Ginkgolides ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; MAP Kinase Signaling System ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; metabolism

Objective CD4 +IL-17 +cells are a group of newly discovered effector CD4 +T cells, which may play a key role in the pathogenesis of cancer.This study aims to investigate the expres-sion of CD4 +IL-17 +cells in pancreatic cancer and its correlation with the clinicopathological characteristics and prognosis of the dis-ease as well as the clinical significance of the cells in the microenvironment of pancreatic cancer. Methods We collected tumor tis-sue and tumor-adjacent normal tissue samples from 51 pancreatic cancer patients.We determined the expressions of CD34 and vascular endothelial growth factor ( VEGF) and measured the proportion of IL-17 +cells in the cancer tissue using immunohistochemistry and the fluorescence activated cell sorter, respectively, followed by analysis of their correlation with tumor angiogenesis, clinicopathological pa-rameters, and survival time of the patients. Results The percentage of CD4 +IL-17 +cells in tumor tissue was positively correlated with microvessel density (r =0.534, P<0.05) and the expression of VEGF in the tumor tissue (r=0.356, P<0.05).IL-17 +cells were expressed more highly in the tumorous than in the tumor-adjacent normal tissue (P<0.05), and the expression level was correla-ted with the stage of tumor, node, and metastasis (TNM) and lymph node metastasis (P<0.05), but not with the patients′gender or age, tumor size, tumor location, histological grade, or local invasion (P>0.05).Fifty (98.0%) of the patients were successfully followed up for 2-67 months, which revealed a median survival time of 16.6 ±4.8 months, significantly longer in those with a higher expression of intratumoral IL-17 +cells (P<0.05).Univariate analysis showed an association of the survival rate with the tumor size, TNM stage, lymph node metastasis, and level of intratumoral IL-17 +cells, while multivariate analysis showed the TNM stage to be an independent prognostic factor for the survival of the pancreatic cancer patients. Conclusion The expression of CD4 +IL-17 +cells in the tumor tissue is positively correlated with tumor angiogenesis, while that of IL-17 +cells with the clinicopathological parameters and survival time of the patients and therefore may serve as an important immune indicator for the prognosis of pancreatic cancer.

The mean lead levels in the maternal blood, cord blood, breast milk and placental tissue, were 0.63 μmol/L (13.2 μg/dL), 0.33 μmol/L (6.90 μg/dL), 4.74 μg/L and 0.86 μmol/kg (17.85 μg/100g) respectively for 165 parturient women occupationally non-exposed to lead in 2 hospitals in Shanghai. No significant difference was found between maternal age groups for these indicators. However, the lead levels in the cord blood and breast milk increased with the lead level in the maternal blood, with coefficient of correlation of 0.714 (P＜0.0001) and 0.353 (P＜0.01) respectively. The mean concentration of lead in breast milk for 12 occupationally lead exposed women was 52.7 μg/L, which was almost 12 times higher than that for the occupationally non-exposed population. These results suggested that transfer of lead via placenta prenatally and breast milk postnatally were possible and might pose a potential health hazard to the fetuses and the neonates.

Objective To analyze the data of specialist counseling on tuberculosis, investigate the patients' demands,and to provide references for supplementing the content of specialized health education and making personalized health education plans. Methods Data of 2877 original telephone consultations and outpatient consultations were retrospectively collected and analyzed. Results The majority of client population were adults aged from 18 to 60 years old, from other provinces or cities, lowly educated (junior high school),married, newly diagnosed workers infected with pulmonary tuberculosis. 31.62% of the reasons of consultation were concern of the transmission of tuberculosis among patients and their families. Their knowledge about tuberculosis mainly came from the network, accounting for 54.01%. The contents of consultation included types of knowledge, information, service and psychological, with the rates of 67.43%, 13. 1%, 13.45% and 6.01%, respectively. 46.76% of the psychological problems were fears of tuberculosis. 98.75% of patients'problems were resolved in time by counseling. Conclusions Based on the analysis of the data, specialist counseling services should be carried out as it has great significance for improving health education and extensive services of tuberculosis patients.

OBJECTIVETo provide the basis for AIDS intervention, the study on the relationship between AIDS related risk behaviors and the related factors was carried out among prostitutes, injection drug users and long-distance truck drivers.

METHODSQuestionnaire investigation and statistical analysis as chi(2) test, F test, logistic regression were adopted to analysis the relationship between AIDS related risk behaviors and the correlated factors.

RESULTSKnowledge about AIDS seemed to be related to their level of understanding the problem (in commercial sex workers r = 0.307, P = 0.000, in injection drug users F = 93.07, P = 0.000, in truck man F = 30.06, P = 0.000). Condom use when entertaining their clients last time was related to the knowledge of HIV transmission in commercial sex workers and truck drivers (OR = 1.171, 1.145) and knowledge of HIV prevention (OR = 1.081, 1.397), in drug users regarding gender difference (OR = 2.121).

CONCLUSIONThis study addressed that the effective means to reduce the rate of AIDS risk behaviors and to lessen the harm of AIDS are to improve the knowledge of AIDS and the effective methods to prevent AIDS in the high risk population.

Acquired Immunodeficiency Syndrome ; prevention & control ; Automobile Driving ; China ; Health Education ; Humans ; Risk-Taking ; Sex Work ; Substance-Related Disorders ; Surveys and Questionnaires