Animals ; Endothelium, Vascular ; Humans ; Lipid Metabolism ; Metabolic Diseases

OBJECTIVE: To provide reference about the way to consummate hospital drug quality management system and ensure drug quality and patients' medication safety.METHODS: Efficient drug maintenance measures were explored by analyzing and summarizing the specific work and conservation procedures of drug maintenance in our hospital which included the humidity of storehouse,monitoring of storage condition,color-code management,expiration date management etc.RESULTS & CONCLUSIONS: Drug maintenance should be aimed at guaranteeing the drug quality;moreover,great importance should be attached to regular quality examination and maintenance of drugs and take timely effective measures aimed at the problems presented so as to ensure the drug quality,safety and effectiveness.

Objective To study the survival, migration and distribution of allogeneic marrow stromal cells (MSCs) which were marked with fluorescence agent Hoechst 33342 in vitro and then transplanted into the mice with acute radiation injury, and to explore the hematopoietic reconstitution in allogeneic MSCs transplantation. Methods The bone marrow of murine femoral bone was washed out with DMEM, the cell suspension was gathered and centrifuged twice, and then the MSCs were cultured in vitro, marked with Hoechst 33342 at final concentration of 10?g/ml. 30 minutes later, the MSCs were gathered and transplanted into radiation injured mice via intravenous injection. Each mouse was injected with 2?105 MSCs. The distribution of marked MSCs was observed in heart, liver, spleen, lung and kidney of radiation injured mice at time points of 1w, 2w and 3w after MSCs transplantation. Results After being marked with Hoechst 33342, the nuclei of MSCs were in bright blue color. All mice of MSCs-transplant-group survived until execution, but three mice of Non-MSCs-transplant-group died during three weeks. Only a few marked MSCs were found in heart, liver, lung and kidney in the frozen section at each time point, but a large number of marked MSCs were found on the surface of spleen, and colonies were formed two weeks after introduction of MSCs. Conclusions Transplanted MSCs can not only survive in the irradiation injured mice, but also specifically migrate to the injured site, to participate in the reconstitution of hematopoiesis after acute radiation injury.

AIM: To identify and quantitative analysis gentiopicroside content in the extract of Violent gentian of Tibetan herbal medicine. METHODS: Application of HPLC-the electricity sprays fog-mass spectroscopy/mass spectroscopy(HPLC-ESI-MS/MS) and the technique of LC-DAD-UV,by contrast with the reservation time of the authentic gentiopicroside,the ultraviolet absorbed spectrum,the molecule~+,quickly identified the gentiopicroside in the extract of violent gentian.As to it's quantitative analyis we adopted the capillary HPLC methods,with the weight of authentic gentiopicroside for abscissa. RESULTS: MS showed that the methanol extract of violet gentian mainly had five peaks.the compound structure of first peak was gentiopicroside,molecule~+ 356.9,result showed that gentiopicroside in gentian was 1.97%. CONCLUSION: Gentiopicroside is the mainly component of violet Gentia.

Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective To determine the clinical use of CM on interventional therapy for middle and late stage cancer.Methods We observed the effect of clinical administration on the patients with interventional therapy or inteventional therapy plus CM,control with themselvies.Results 38(76%) out of 50 patients got better than that of preadministration,cancer mass decreased.Conclusion CM can increase the sensitivity of carcinoma cells to chemicals and decrease its sideeffect.

Objective To investigate the feasibility of serum cardiac troponin T(cTnT) as a marker of cardiovascular events and hemodialysis (HD) adequacy in maintainence hemodialysis (MHD) patients.Methods Forty-seven cases of MHD patients were randomly divided into two groups (group A and group B).Group A received intermittent HD 4 h thrice one week,and group B received intermittent HD 4 h twice one week plus high-flux hemodiafiltration(HDF) 4 h once one week.Serum examination for blood biochemical indicator,cTnT and echocardiogram was performed every three months and at the time of recruitment.All the patients were followed up until the occurrence of death or cardiovascular events.Results After 3 months treatment,serum cTnT deceased significantly in group B compared with group A,and maintained the lower levels throughout the follow-up.E/A and LVEF had been reduced since 3 months treatment in group A,but stable in group B,E/A was lower in group A after 18 months treatment than that in group B,LVEF was lower in group A after 12 months treatment than that in group B.There were positive correlations between cTnT and E/A or LVEF in 42 cases who accomplished the follow-up of 12 months (r =0.54,0.66,P ＜0.05).Kaplan-Meier survival curve showed that the occurrence of cardiovascular events in patients with cTnT≥0.1μ g/L was higher than that with cTnT ＜0.1 μg/L in (28.5 ± 9.7) months' follow- up (Log-rank test: P =0.02).Both survival analysis and Cox analysis indicated that serum cTnT was a predictor of cardiovascular events in MHD patients.Conclusions Serum level of cTnT can be used as a marker of HD adequacy,and it is a predictor of cardiac events in MHD patients.Regular high-flux HDF increases the adequacy of HD treatment and improves the quality of life in MHD patients.

Objective To investigate the effect of electroacupuncture at point Fenglong(ST40) on the expressions of macrophage surface antigen CD11b and intercellular adhesion molecule-1 (ICAM-1) in hyperlipidemia rats. Method Forty healthy SD rats were randomly allocated to normal control, high fat diet, high fat+common diet, high fat diet and treatment, and high fat+common diet and treatment groups. After 28 days of treatment with electroacupuncture at point Fenglong, blood lipid levels, namely, total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) contents were measured in every group of rats. The expressions of peritoneal macrophage surface antigen CD11b and ICAM-1 were examined using flow cytometry (FCM) in every group of rats. Result Plasma TC and LDL-C increased significantly in the high fat diet group of rats compared with the normal control group (P<0.01). In the high fat+common diet group of rats, serum TC and LDL-C decreased significantly in comparison with the high fat diet group (P<0.01) but increased significantly in comparison with the normal control group (P<0.01). After treatment with electroacupuncture at point Fenglong, rat plasma TC and LDL-C decreased significantly (P<0.01) and TG and HDL-C did not change significantly (P>0.05). The expression rates of macrophage CD11b and ICAM-1 increased significantly in the high fat diet group of rats compared with the normal group (P<0.01). In the high fat+common diet group of rats, the expression rates of macrophage CD11b and ICAM-1 decreased significantly in comparison with the high fat diet group (P<0.05) but increased significantly in comparison with the normal group (P<0.01). The expression rates of CD11b and ICAM-1 decreased significantly in the high fat diet and treatment group compared with the high fat diet group (P<0.01) and decreased significantly in the high fat+common diet and treatment group compared with the high fat+common diet group (P<0.01) and the high fat diet and treatment group (P<0.01). Correlation analysis showed that CD11b and ICAM-1 levels had a significantly positive correlation (r=0.947, P<0.01). Conclusion Electroacupuncture at point Fenglong can significantly down-regulate blood fat TC and LDL-C levels and the expressions of macrophage CD11b and ICAM-1 in hyperlipidemia rats. It has a certain therapeutic effect on hyperlipidemia.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

Objective To study the feasibility of TNF-α promoting migration of rat mesenchymal stem cells (MSCs) to local damaged tissues. Methods The MSCs was exposed to TNF-α at different concentrations and the expression rate of surface adheslon molecules and specific markers as well as their adhesion to endothelial cells were detected.Based on the above steps,the MSCs stimulated with the optimal concentration of TNF-α were obtained and were injected intravenously to the rats whose hindlimbs experienced ischemia damage.The rats were executed for achieving the muscle samples in the ischemic area,which were made into frozen section to count the number of MSCs. Results ( 1 ) Twenty-four hours after the TNF-o stimulation,the expression of adhesion molecule (VCAM-1) of MSCs increased in a concentration-dependent manner,while the expression of adhesion molecules (ICAM-1,L-Selectin and VLA4) of MSCs showed no significant changes.Besides,the expression rate of specific markers of MSCs was also obscure.(2) Exposed to 10 ng/ml TNF-o,MSCs presented an obviously increased ability in adhesion to the endothelial cells.(3) MSCs stimulated with 10 ng/ml TNF-α showed a larger number in the ischemia-damaged tissue of rat hindlimbs than that in the control group. Conclusion TNF-α at concentration of 10 ng/ml is effective within a short term in increasing VCAM-1 expression in rat MSCs and promoting the adhesion of MSCs to endothelial cells without affecting their character.