Objective Normal estimation is the key step for volume visualization. Commonly used methods for normal estimation are based on interpolation and derivative. A novel normal estimation algorithm based on approximation for visualization of medical images was presented in this paper. Method It approximated the density function in local neighborhood with a second-degree polynomial function. The coefficients of the polynomial function were solved by minimizing the error of the approximation and the gradient vector at arbitrary point was obtained directly from the analytical derivative of the density function without interpolation. Because of symmetry, the solution of this equation was simplified.This method was tested in several volume data sets. The results and the generation time by different methods were obtained and compared. Result The results showed that this algorithm produced satisfactory quality images while the computational complexity was not increased. Conclusion This approach is preferable for most applications, especially for medical images reconstruction.

OBJECTIVETo observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.

METHODSNB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.

RESULTSMAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).

CONCLUSIONMAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.

Alkaloids ; Antineoplastic Agents ; Cell Differentiation ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Phospholipid Transfer Proteins ; metabolism ; Quinolizines ; RNA, Messenger ; Signal Transduction ; Tretinoin ; Tumor Cells, Cultured ; Up-Regulation

Objective To investigate the cytotoxicity of vascular endothelial growth factor (VEGF) siRNA combined with fusion suicide gene yCDglyTK on human gastric cancer cell line in vitro.MethodsThe gastric cancer cell line SGC7901 was transfeeted with blank plasmid pcDNA3.1 (-) null [pcDNA3.1 (-) group], or VEGF-siRNA expression plasmid pGenesil-shVEGF (SGC7901/shVEGF group),or fusion suicide gene plasmid pcDNA3.1 (-)CV-yCDglyTK (SGC7901/CDTK group),or combined gene plasmid pcDNA3.1 (-)shVEGF-yCDglyTK (SGC7901//shVEGF-CDTK group) with calcium phosphate nanoparticles (CPNPs).Un-transfected gastric cells were set as control group.The stable transfected cells were selected by G418.The target gene expression was verified by RT-PCR and Western-blot.After given prodrug 5-fluorocytosine (5-FC), the biologic characters variation, apoptotic morphology and apoptotic rate of cells in each group were observed through cell growth curve by MTT assays, by-stander effect, Hoechst 33258 staining and flow cytometry.The data was analyzed with SPSS 13.0 software and multiple groups’ comparison was analyzed with LSD test.ResultsFour gastric cancer cells lines transfected with different plasmids were successfully established.The expression of gene yCDglyTK was detected both in SGC7901/CDTK cells and SGC7901/shVEGF-CDTK cells.By MTT assays, the cell growth curve indicated that the A570 value of SGC7901/shVEGF cells, SGC7901/CDTK cells and SGC7901/shVEGF-CDTK cells decreased significantly compared with that of SGC7901 and SGC7901/null cells after a 24-hour 5-FC treatment (P＜0.01).When the percentage of stable gene trasfected SGC7901 cells was 60％, 80％and 100％, the cell relative viability was 13.09％±2.40％, 9.74％±2.83％ and 5.68％±1.03％,respectively. A large number of cells in SGC7901/CDTK and SGC7901/shVEGF-CDTK group appeared typical apoptotic morphology under fluorescence microscope.The result of flow cytometry showed that the apoptosis rates in SGC7901/shVEG group、 SGC7901/CDTK group and SGC7901/shVEGF-CDTK group were 16.40％ ±4.68％, 57.63％ ± 4.96％ and 69.07％ ± 4.69％,respectively, and there were significant differences compared with control (P＜0.01).Conclusion VEGF siRNA combined with suicide gene can effectively kill gastric cancer cell line SGC7901.Apoptosis induction may be one of the important mechanisms of killing tumor cells.

AIM: To study the effect of ginkgolide B on high glucose-induced human lens epithelial cell (HLEC) apoptosis.
METHODS: The high glucose-induced HLEC model was established. Different concentrations of ginkgolide B were intervened. Cell viability was assayed by MTT assay, morphology of cell apoptosis was observed by hochest33258 staining. Cell ultrastructural changes were detected by transmission electron microscopy. Expressions of apoptosis - related factors caspses - 3 and caspase-9 were tested by colorimetric detection.
RESULTS: High glucose inhibited the activity of HLECs, induced apoptosis reaction of HLECs, caused high expression of apoptosis factors in cells; while ginkgolide B inhibited the decrease of cell viability induced by high glucose, decreased the HLEC apoptosis and reduced the expression of apoptosis factors Caspase-3 and Caspase-9.CONCLUSION: Ginkgolide B may inhibit the expression of caspses-3 and caspase-9 then effectively inhibited high glucose-induced apoptosis in HLECs.

OBJECTIVETo compare the effect of reinforcing Shen method (RSM) and activating blood method (ABM) in treating osteoarthritis (OA) at the molecular level.

METHODSThe physical and chemical characteristics of components from respective recipes of RSM and ABM, and network features of component-target interaction network were analyzed by computer simulation methods including chemical space, molecular docking, and biological network, etc.

RESULTSThe chemical components of RSM and ABM were scarcely scattered with larger overlapping. Among established networks, the distribution of network features was partially similar in RSM and ABM. The average target number correlated with each component was 1.86 in RSM and 2.11 in ABM respectively. Each average target number was respectively correlated with 4.46 compounds and 3.93 compounds, reflecting multi-component and multi-target actions.

CONCLUSIONComputer simulation could intuitively trace out similarities and differences of two different methods and their interaction with targets, which revealed that the compatibility of RSM and ABM could have broader protein targets and potential synergism at the molecular level.

Computer Simulation ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Osteoarthritis ; drug therapy ; Phytotherapy ; methods

OBJECTIVETo investigate the molecular mechanism of haemophilia B caused by the novel mutation of Arg327Ile (R327I) in FIX gene.

METHODSThe R327I, R327Ala(A), R327Lys(K), R327Asn(N) and a replacement mutant (FIXβFVII), in which FIX β strand 324-329 was replaced by that of FVII 298-303, expression plasmids were constructed with site-directed mutagenesis method based on the wild-type (WT) FIX expression plasmid. The HEK293 cell was transiently transfected, then the activity of FIX (FIX:C) was assayed by one stage method in the conditioned medium, while the FIX:Ag in both the conditioned media and the cell lysates was measured by ELISA. The molecular weight and the semi-quantity of expressed FIX were analyzed by Western blot. Fluorescent protein expression plasmid was constructed to investigate the synthesis and secretion of the FIX R327I mutation in the viable cells.

RESULTSFIX:C of the R327I mutant protein was 4.49% of the level of the WT in the conditioned medium, and the FIX:Ag of the R327I mutant protein in the conditioned medium and the cell lysates was 31.02% and 129.29% compared to that of WT, respectively. The mutation was characterized as cross-reaction material reduced (CRMR). The viable cell fluorescent assays showed that the R327I protein was more in both the viable cells and in lysosome than that of WT. The FIX:C of the R327A, R327K, R327N and FIXβFVII mutants was reduced compared to that of WT, the reduction of FIX:C of FIXβFVII was the most significantly amount among all the mutants in medium. FIX:Ag of all the mutants in the medium, except that the R327K increased, was reduced. The result of Western blot showed that the molecular weight of R327I protein was the same as that of WT, but the amount of the protein was much less compared with WT in the conditioned medium.

CONCLUSIONThe abnormal synthesis and secretion as well as the abnormal function of the R327I mutant protein causes haemophilia B. The residue of R327 as well as the β strand domain of R327 located play important roles of the specific function of FIX.

Factor IX ; genetics ; HEK293 Cells ; Hemophilia B ; genetics ; pathology ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Transfection

OBJECTIVETo evaluate the effectiveness of a school-based smoking prevention and control intervention program among elementary school students.

METHODSThrough two phase cluster sampling, 566 pupils in grade 4 and grade 5 of two schools were assigned to intervention group and control group. One year comprehensive smoking intervention was conducted in the intervention group. The assessment was carried out through three questionnaires: pre- and post-intervention, 6-month after intervention.

RESULTSAfter one year intervention, pupils in the intervention group significantly improved their knowledge and attitudes related to tobacco use. The rate of attempting smoking decreased form 7.8% to 2.6% and the rate of passive smoking from 53.6% to 41.8%. The difference between the intervention and control groups was statistically significant. However, several index started to decline at 6-month follow up survey.

CONCLUSIONThe result demonstrated the effectiveness and feasibility of tobacco control in elementary school and the positive effect must be developed.

Case-Control Studies ; Child ; China ; epidemiology ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Schools ; statistics & numerical data ; Smoking ; epidemiology ; Smoking Prevention ; Surveys and Questionnaires ; Tobacco Smoke Pollution ; statistics & numerical data

OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

To observe the protective effect and mechanism of Sailuotong capsule in focal cerebral ischemia/reperfusion. The 90 min middle cerebral artery occlusion (MCAO) reperfusion model was established. The expressions of dynamin-related protein 1 ( Drp1) and optic atrophy 1 (Opa1) were tested by Western blot. The transmission electron microscope was used to observe the changes in the mitochondrial ultra-structure. The pathological morphological changes were observed through the HE staining. The immunohistochemical method was used to test Drp1 and Opa1 expressions. Sailuotong capsule (33, 16.5 mg x kg(-1), ig) can inhibit the abnormal mitochondrial fission and fusion in the cortical area on the ischemia side and the mitochondrial fission gene expression and promote the mitochondrial fusion gene Opa1 expression, so as to alleviate the energy metabolism disorder caused by ischemia/reperfusion. Sailuotong capsule can inhibit the abnormal mitochondrial dynamics in peri-ischemic regions and maintain the normal morphology of mitochondria, which may be the mechanism of Sailuotong capsule in promoting the self-recovery function in the ischemic brain region.
Animals ; Brain ; drug effects ; metabolism ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; surgery ; Drugs, Chinese Herbal ; administration & dosage ; Dynamins ; genetics ; metabolism ; GTP Phosphohydrolases ; genetics ; metabolism ; Humans ; Male ; Mitochondria ; drug effects ; metabolism ; Rats

OBJECTIVETo evaluate the clinical efficacy of treating myelodysplastic syndrome (MDS) by hematopoietic stem cell transplantation (HSCT) combined with Chinese medical syndrome typing.

METHODSFrom July 2009 to July 2013, 6 MDS patients were treated with allo-HSCT combined with Chinese medical syndrome typing from HLA-identical sibling donors at Department of Hematology, Zhejiang Provincial Hospital of Chinese Medicine. Patients were classified as refractory anemia (RA, 2 cases), refractory anemia with ringed sideroblast (RARS, 1 case), refractory cytopenia with multilineage dysplasia (RCMD, 2 cases), and RA with excess blasts-I (RAEB-I , 1 case). Modified BuCy conditioning regimen was used in all 6 cases. Two patients received bone marrow transplantation (BMT), 1 patient received peripheral blood stem cell transplantation (PBSCT), and 3 patients received BMT + PBSCT. In order to prevent the occurrence of graft-versus-host disease (GVHD), all patients were treated with cyclosporine + methotrexate + mycophenolate mofetil. Different Chinese medical treatment methods (by syndrome typing) were given to patients according to different criticality of international prognostic scoring system (IPSS, 5 at moderate risk and 1 at high risk).

RESULTSAll 6 patients successfully reconstructed their hematopoietic system. The time from transplantation to ANC ≥ 0.5 x 10(9)/L and platelet (PLT) ≥ 20 x10(9)/L were 13 (9-15) days and 11 (9-22) days respectively. Main complications were GVHD. Acute GVHD (aGVHD) occurred in 4 cases, 3 cases of grade I and 1 case of grade II, and local chronic GVHD (cGVHD) occurred in 1 patient. All cases survived with median follow-up of 18 (11-58) months. The overall survival (OS) and disease-free survival (DFS) rate were 100%.

CONCLUSIONSHSCT combined with Chinese medical syndrome typing could improve clinical symptoms, reduce transplant as- sociated complications. So it was an effective treatment choice for MDS.

Biomedical Research ; Blood Platelets ; Bone Marrow Transplantation ; Cyclosporine ; therapeutic use ; Disease-Free Survival ; Drugs, Chinese Herbal ; therapeutic use ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Humans ; Medicine, Chinese Traditional ; Methotrexate ; therapeutic use ; Myelodysplastic Syndromes ; therapy ; Transplantation Conditioning ; Transplantation, Homologous ; Treatment Outcome