Objective To investigate the methylation status in the promoter region of Dickkopf-3 (Dkk3) gene in patients with myelodysplastic syndromes (MDS),and to initially explore the relationship between the methylation of this gene and survival time.Methods Methylation-specific PCR (MSP) was applied to measure the promoter methylation of Dkk3 gene in 43 bone marrow or peripheral blood samples of MDS patients.As controls,70 normal peripheral blood samples from general outpatients were examined.Results In 43 patients with MDS,7 patients (16.3 ％) showed Dkk3 gene methylation.And 5 of them were semi-methylation status,2 of them were exhaustive methylation status.In 70 controls,1 showed Dkk3 gene semi-methylation.The frequency of methylation in MDS patients was significantly higher than that of controls (x2 =8.93,P =0.005).In the Dkk3 methylation group,2/7 were from bone marrow and 5/7 were from peripheral blood.Meanwhile,2 patients were RA,1 patient was RCMD,4 patients were RAEB.There was no significant difference between the different sample source (bone marrow or peripheral blood) for the results of the methylation status (x2 =0.051,P =0.821).Either between the different sex,age,type,chromosome and WPSS score (P ＞ 0.05).The progress of disease didn't influence the methylation frequency (P ＞ 0.05).The smvival analysis showed no relationship between the methylation of this gene and smvival time.Conclusions In this MDS group,there is high level of methyl-modification in Dkk3 gene.The methylation of Dkk3 might be one of the molecular mechanisms that contribute to the progress of patients with MDS.The peripheral blood sample maybe a better substitute in detective of Dkk3 with MDS.

With the rapid economic growth and social development in the BRICS countries, the role of devel-opment assistance for health is becoming more and more significant. This paper describes the scales, recipient coun-tries, mechanisms and characteristics and management systems of development assistance for health in BRICS coun-tries. The paper suggests that a) it is necessary to set up a centralized international aid management agency;b) the mode of development assistance for health must be optimized;c) the scale of development assistance for health shall be increased ( over time);d) each BRIC country should use its own comparative advantages and development experi-ence to carry out development assistance for health while strengthening the cooperative power among the BRICS coun-tries;e) development assistance for health data should be more transparent and open;f) the evaluation of develop-ment assistance for health must be established and perfected.

Objective: To discuss the pharmaceutical care experience of clinical pharmacists in the antibacterial agents application in one patient with infection after coronary artery bypass graft ( CABG) . Methods:The clinical pharmacist participated in the treatment of the patient after CABG. According to the relevant laboratory indices and the extent of the infection combined with the vital signs of the patient, the pharmacist formulated and adjusted the anti-infection therapeutic regimen. The therapeutic effects and adverse reactions were observed as well. Results: Safte and high-quality individualized pharmaceutical service was provided to the patient by the pharmaceutical care of the clinical pharmacist in anti-infection treatment. Conclusion:Through the above practice, clinical pharmacists have played a positive role in reasonable using of anti-infective drugs in patients with cardiac surgery.

Objective:To investigate variety pattern of expression level of TCRV?subfamilies in mononuclear cell in spleen tissue of rats with dampness pathogenic factors and normal rats by using FQ-PCR technique. Methods:32 SD rats were divided into four groups: normal group, external dampness group, internal dampness group, external and internal dampness group. Observing period was 20 days. 3 Rats were randomly selected from each group in order to exam the TCRV? subfamilies expression level. Results:The expression of TCRV?1, TCRV?7, TCRV?9 and TCRV?13 in external dampness group were higher than those in normal group (P

Objective To explore the imageologic characteristics of pelvic fractures with artery injuries and the treatment methods for embolization of arteries.Methods From January 1999 to June 2005,60 cases(42 males and 18 females)aged 21-52 years(average 34.5 years)with pelvic fractures and unsteady blood dynamics were admitted into our hospital.There were 32 cases with traffic injury,13 with crushing injury,nine with fall injury and six with other injuries.The mean injury severity score was 39?16(16-66).All cases were hypetensive with systolic blood pressure less than 90 mm Hg on the arrival.Routine X-ray examination of dorsaventral,debouch and porch of pelvis was performed.The aver- age amount of blood transfusion was 2 886 ml.All cases underwent iliac artery angiography and pelvic ar- teriography.Results X-ray examination of pelvic fractures showed posterior pelvic fracture in 25 ca- ses,with 64 branches of blood vessels injured;anterior pelvic fracture in 13,with 17 branches of blood vessels injured;acetabular fracture in six,with 12 branches of blood vessels injured;and combined pel- vic fracture in 16,with 36 branches of blood vessels injured.Three cases died,with mortality rate of 5%.One case with common arterial thrombosis was treated with artificial blood vessel transplantation, four cases with external iliac artery injuries including one with artery rupture were treated with prosthesis, and among the three cases with external iliac artery thrombosis,one was treated with dislodgment of thrombosis,one treated with recanalization of thrombolysis and one did not give any treatment.Fifty cases with injury and bleeding of internal iliac artery and its branches were treated with arterial embolization. Five cases showed no obvious injury.Conclusions The types of artery injuries can be predicted through X-ray of pelvic fracture.Posterior pelvic fracture may easily cause injury to superior gluteal arter- ies,iliac lumber arteries,and lateral sacral arteries.While anterior pelvic fracture will cause injury to obturator arteries.Superior gluteal artery is susceptible to injury.Embolization of injured arteries and an- astomosis are preferred treatment for pelvic arterial disruptions.

ObjectiveTo investigate the methylation status in the promoter region of secreting frizzled related protein 2 (SFRP2) gene in patients with myelodyplastic sydrome (MDS) and to initially explore the relationship between the methylation of this gene and prognosis/survival time.MethodsMSP method was applied to examine the promoter methylation of SFRP2 gene in 43 bone marrow or peripheral blood samples of MDS patients.As controls,70 normal peripheral blood samples from volunteers of general outpatients were examined.Then some of the patients were followed up.ResultsIn 43 patients of MDS,10 samples (23.3 ％)showed SFRP2 gene methylation,and all of them were semi-methylation status.In 70 controls,no sample showed SFRP2 gene methylation.The frequency of SFRP2 gene methylation in MDS patients was significantly higher than that in controls (x2 =17.86,P ＜0.0001).Of the 10 SFRP2 gene methylation samples,5 were bone marrow samples and 5 were peripheral blood samples.In this group of patients,3 patients were diagnosed as RA,1 patient was diagnosed as RAS,2 patients were diagnosed as RCMD,3 patients were diagnosed as RAEB and 1 patient was diagnosed as MDS-U.There was no significant difference between the different sample source (bone marrow or peripheral blood) for the results of the methylation status (x2 =0.912,P ＞0.05).Either no significant difference between the different sex,age,type,chromosome and WPSS score (all P ＞0.05).The progress of disease didn' t influence the methylation rate (P ＞0.05).16 patients accepted follow-up and 11patients died,3 patients went to AML.2 died patients showed SFRP2 gene methylation.The survival analyses showed no relationship between the methylation of this gene and survival time(x2 =0.022, P ＞0.05).ConclusionIn this MDS group,there is a high level of methyl-modification in SFRP2 gene.The methylation of SFRP2 may be one of the molecular mechanisms that contribute to the progress of patients with MDS.The peripheral blood sample maybe a better substitute in detection of SFRP2 with MDS.

Clinical study of modified Ganmai Dazao decoction in the treatment of yang deficiency climacteric depression and observe the effects of modified Ganmai Dazao decoction on neuroendocrine system in patients with yang deficiency climacteric depression. 86 cases were randomly divided into treatment group treated with modified Ganmai Dazao decoction and control group treated with Deanxit. The curative effect was evaluated with Hamilton's depressive scale (HAMD) and pittsburgh sleep quality scale (PSQI) before and at the end of the two and four weeks of the treatment, the serum levels of serotonin (5-HT), norepinephrine (NE), estradiol (E2), follicle stimulating hormone (FSH), luteotropic hormone (LH) were detected before and after the four weeks of the treatment The results showed that the total effective power of treatment group was 88.4% and the total effective power of control group was 81.4% after four weeks interference, with insignificant difference between the two groups. After two and four weeks of the treatment, the score of HAMD decreased remarkably in both groups (P < 0.01), with insignificant difference between the two groups in same phase. After two and four weeks of the treatment, the total score of PSQI decreased remarkably in both groups (P < 0.05), with significant difference between the two groups after four weeks (P < 0.01). After four weeks of treatment, the serum levels of 5-HT and NE increased (P < 0.01), with insignificant difference between the groups. After four weeks of treatment, the serum levels of E2 increased obviously (P < 0.05), the levels of FSH decreased obviously (P < 0.05), the levels of LH decreased insignificant, with insignificant difference between two groups. This study indicates that modified Ganmai Dazao decoction has obvious therapeutic effects in the treatment of climacteric depression, and showed equivalent efficacy with Deanxit, and modified Ganmai Dazao decoction has better effect on improving the sleep quality in patients than Deanxit, the effect of improved clinical symptoms may be through adjusted levels of 5-HT, NE, E2, FSH and LH of climacteric depression.
Adult ; Depression ; blood ; drug therapy ; etiology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Menopause ; drug effects ; psychology ; Middle Aged ; Neurosecretory Systems ; drug effects ; metabolism ; Norepinephrine ; blood ; Phytotherapy ; Serotonin ; blood

Objective To construct a bicistronic expression vector containing HPV type 6b L1 gene, to express the recombinant vector in mammalian cells, and to establish a cell strain stably expressing HPV6b L1 gene. Methods After endonuclease digestion and purification, the gene fragment of HPV6b L1 was cloned into the eukaryotic expression vector pIRES2-enhanced green fluorescent protein (EGFP). The identification of the recombinant was realized via endonuclease digestion and sequence analysis. Then, the recombinant plasmid pIRES2-HPV6bLl-EGFP was transfected into NIH3T3 (a mouse embryonic fibroblast cell line) cells. Subsequently, the expression of EGFP was observed by fluorescent inverted microscopy, and HPV6b L1 mRNA expression by reverse transcription (RT)-PCR. Results The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed, transfected into N1H3T3 cells, and selected by G418. The expression of EGFP was seen under an inverted fluorescence microscoy. RT-PCR proved the expression of HPV6b LI mRNA in transfected cells. Conclusions The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed and transfected into NIH3T3 cells. Inverted fluorescent microscopy and RT-PCR confirmed the successful expression of HPV6b L1 in NIH3T3 cells.

In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 μm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum
Acetonitriles ; Chromatography, High Pressure Liquid ; Flavonoids ; chemistry ; Kaempferols ; Quercetin ; analogs & derivatives ; Rutin ; Tandem Mass Spectrometry ; Vitaceae ; chemistry

OBJECTIVETo evaluate the effects and molecular mechanism of action of Changweiqing (CWQ) in reversing multidrug resistance by observing its impacts on nuclear translocation of Y-box binding protein-1 (YB-1), multi-drug resistance gene (MDR1) expression and P-glycoprotein (P-gp) expression in human colon cancer cell line HCT8/V with drug-resistance induced by vincristine.

METHODSCultured HCT8/V cells were exposed to the CWQ-containing rat serum prepared by drug perfusion. YB-1 expressions in cell plasma and nuclei were examined by Western blot; the binding activity of YB-1 to MDR1 gene promoter sequences was detected by electrophoretic mobility shift assay (EMSA); the mRNA transcription levels of MDR1, YB-1 and multi-resistance related protein (MRP) were examined by RT-PCR; the expression of P-gp on cell membrane was determined by flow cytometry. Results Along with the increasing drug's concentration of CWQ-containing serum from 1.25% up to 2.5% and 5%, the expressions of YB-1 decreased in HCT8/V cell nuclear and increased in cytoplasm gradually; the binding activity of YB-1 to MDR1 gene promoter weakened (P < 0.01), MDR1 mRNA expression and fluorescence intensity of P-gp on cell membrane attenuated (P < 0.05 or P < 0.01), while YB-1 and MRP mRNA unchanged (P > 0.05).

CONCLUSIONCWQ could reverse the drug-resistance of colon cancer cells by influencing nuclear translocation of YB-1 and reducing the expression of MDR1/P-gp.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Active Transport, Cell Nucleus ; drug effects ; Animals ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Rats ; Vincristine ; pharmacology ; Y-Box-Binding Protein 1 ; metabolism