AIM:To seek a simple and reliable process for extracting effective components in Diedahuoxue film.METHODS: According to uniform design, five factors in extraction were optimized, i.e. ethanol concentration, reflux time, reflux times,ethanol volume and drug particle. The absorption value was taken as evaluating parameter in UV- spectrophotometry. RESULTS: The results showed that the best extracting conditions were 95% ethanol, ten times amount of ethanol, 0.2mm size of particle and reflux time of 1h with 4 repetitions.CONCLUSION:This process is simple and reliable.

Objective To explore effects of human embryonic stem cells ( hESCs) on proliferation, invasion and migration of SK-Hep1 human hepatoma cells in the co-culture of micro environmen of hESCs and SK-Hep1 . Methods Single cultured SK-Hep1 cells were served as control group while SK-Hep1 which non-contact co-cul-tured with hESCs was regarded as experimental group .The proliferation ability of SK-Hep1 was measured by MTT method; invasion and migration ability of SK-Hep1 cells were detected by Transwell chamber method;the nucle-us variation and cell apoptosis of SK-Hep1 were detected by Hoechst33258 chromosome and flow cytometry. Results The proliferation of SK-Hep1 cells in the experimental group was obviously inhibited as compared with control group ( P<0.05 );the number of SK-Hep1 cells which passed through the Transwell chambers were sig-nificantly reduced as compared with control group in invasion and migration experiment ( P <0.05 ); more nucleus pycnosis and deformation appeared in experimental group than that in control group .And apoptosis rate of SK-Hep1 cells in the experimental group was significantly higher than that of in the control group ( P<0.05 ) .Conclusions Human embryonic stem cells have inhibitory effect on human hepatoma cell line SK-Hep1 .

Objective To investigate the protective effects of MicroRNA-214 on myocardial injury induced by myocardial ischemia and reperfusion,as well as the regulation mechanism of PI3K and its downstream protein kinase B(AKT)and FoxO1(PI3K / AKT / FoxO1).Methods Wistar rats were randomly divided into 4 groups:sham operation group(Sham group),myocardial ischemia reperfusion injury(IRI)group(IRI group),microRNA-214+sham operation group(MS group),microRNA-214+IRI group(MI group),(n=10,each).The cardiac function was detected at 6 h after ischemia-reperfusion operation.And blood lactate dehydrogenase(LDH),creatine kinase(CK),creatine phosphate kinase isoforms MB(CK-MB),cardiac troponin T(cTnT),serum B natriuretic peptide(pro-BNP)in plasma were detected by enzyme-linked immunosorbent assay(ELISA).Interleukin 10(IL-10),Interleukin 6(IL-6)and tumor necrosis factor α(TNF-α)were assayed.Pathological changes of myocardial tissue were detected by HE and Masson.The expression of microRNA-214 was detected by RT-PCR.The expression of Bax,Caspase-3,BCl2,PI3K,Akt,FoxO1 was detected by Western Blot.Results Compared with Sham group,IRI group showed a significantly increases in myocardial injury parameters of LDH,CK,IL-6 and TNF concentration in plasma,and a significantly reduced concentration of IL-10(P<0.05).And compared with Sham group,MI group showed a significantly increased expression of microRNA-214(P<0.05)and showed a significantly increased myocardial parameters of Bax,Caspase-3,PI3K,Akt protein,and a decreased level of BCl2,FoxO1(P<0.05).Compared with IRI group,microRNA-21 group showed a reduced myocardial ischemia-reperfusion-induced myocardial injury in rats and a reduced plasma concentration of LDH,CK,IL-6 and TNF-alpha,a inhibited expression of caspase-3,Bax,myocardial PI3K and Akt,and a promoted expression of BCl2 and FoxO1 protein(P<0.05).Conclusions MicroRNA-214 reduces the myocardial injury induced by myocardial ischemia-reperfusion through PI3K/Akt signaling pathway.

Objective To observe the clinical effect of rosuvastatin in the treatment of lower extremity atherosclerotic disease in the elderly,and to provide clinical basis of the application of statins in lower extremity atherosclerotic disease in the elderly.Methods 90 patients diagnosed as lower extremity atherosclerotic disease (aged 60 years or over)were randomly divided into simvastatin group and rosuvastatin group (n=45),and they were treated with simvastatin and rosuvastatin,respectively,based on the regular treatment.The total cholesterol (TC),low density lipoprotein-cholesterol (LDL-C), and high sensitivity C-reactive protein (hs-CRP)of the patients in two groups were detected at the begining of treatment and the ending of 32 weeks;at the same time,the lower limb artery intima thickness, strength of hardened, plaque, and artery stenosis degree scores were compared.Results 32 weeks after treatment,the levels of TC,LDL-C and hs-CRP of the patients in two groups were decreased statistically compared with before treatment (P <0.05);the levels of TC,LDC-C,and hs-CRP in rosuvastatin group were lower than those in simvastatin group (P < 0.05 ). 32 weeks after treatment in two groups,compared with before treatment,the lining thickness and plaque,stenosis scores were decreased (P <0.05);the lining thickness,plaque and stenosis scores in rosuvastatin group were lower than those in simvastatin group (P <0.05);the gardening strength grades had no statistical difference between before and after treatment. Conclusion Statins is lipid-lowering and resistant to lower limb arteriosclerosis,and rosuvastatin is better than simvastatin.

Objective To detect the influence of fluoride on the expression TM9SF1 mRNA and Ras mRNA of osteoblasts. Methods The third generation of primary cultured osteoblasts were exposed to a series concentrations of 0,2.5,5.0, 10.0,20.0 mg/L fluoride for 10 days. The influence of different doses of fluorine on the expression of TM9SF1 mRNA and Ras mRNA of osteoblasts cultured in vitro was investigated by SYBR Green I methods. Results The osteoblasts of the control group and the 2.5 mg/L group were in the shape of long spindle, triangle or irregular polygon and had processes, and the cytoplasm was translucent, adjacent cells affixed to each other under light microscope. Those of the 20.0 mg/L group shaped as long spindle or irregular polygon, and some vacuolization and granular materials appeared in cytoplasm. The number of the cells decreased and the volume increased significantly. After exposed to fluoride for 10 days, osteoblasts of 2.5 mg/L group morphologically proliferated. There were statistical siguificances between each groups of TM9SF1 mRNA in human osteoblasts(F = 322.82, P < 0.01). The highest in the 2.5 mg/L group(9326.0 ± 115.97), the expression of TM9SF1 mRNA decreased along with the increasing dose of fluorine. There were statistical significances between 5.0, 10.0,20.0 mg/L groups(6495.0 ± 323.9, 4387.5 ± 545.2, 5962.5 ± 536.7) and control group(9221.0 ± 107.5, all P< 0.01). There was a statistical significance between each groups of Ras mRNA in human osteoblasts(F = 703.28, P < 0.01). The highest in the control group, the expression of Ras mRNA decreased along with the increasing of dose of fluorine. There were statistical significance between 2.5, 5.0, 10.0, 20.0 mg/L groups(6144.5 ± 270.82,5603.5 ± 88.39,3181.0 ± 159.81,4067.5 ± 37.4) and control group(6571.0 ± 196.58). Conclusion The influence on TM9SFI mRNA and Ras mRNA expression in osteoblasts correlates with the dose of fluorine.

italic>Alangium chinense is a commonly used medicinal plant of Alangiaceae Alangium, one of the characteristic Miao medicines in Guizhou. The genus is strongly differentiated and has complex morphological variation, with significant differences in active ingredients and potency among herbs. In this paper, we sequenced the chloroplast genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib using Illumina high-throughput sequencing technology. The genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplasts were sequenced, their assembly annotation and structural characterization were completed, and the genomes of the congeners Alangium chinense and Alangium alpinum were downloaded from NCBI for comparative analysis. Finally, the phylogenetic tree was constructed by selecting published species with close affinity to Alangium chinense family from NCBI. The results were as follows: Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplast genomes were 156 670, 156 672 and 156 871 bp in length, with a typical four-segment structure, all containing one long single copy region (LSC), one short single copy region (SSC) and two inverted repeat regions (IRa and IRb). All of them were annotated to 133 genes, including 88 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Three types of long repeat sequences and SSR loci, forward repeats, palindrome repeats and reverse repeats, were found in all chloroplast genomes. Comparative genomic analysis showed that there was diversity in the boundary transition regions of the five species, no rearrangements or inversions were found in the chloroplast genome, and there were significant differences in the coding regions of ndhA, ycf1, rpl16, ycf2, and petD genes, which provided new loci for molecular identification of Patagonia. In the phylogenetic analysis, Alangium kurzii var. kurzii clustered as a single species, and Alangium chinense subsp. Pauciflorum and Alangium chinense subsp. Strigosum clustered as a single species, with a support rate of 93 and close affinity, indicating that the phylogenetic tree can be used for the species identification. This paper can provide a basis for the taxonomic identification of Alangium, ensure the safety of clinical use of anise herbs and regulate the market of Alangium chinense herbs.

Objective:To study the inhibitory effect of human embryonic stem cells on the HepG2 cells in vitro.Methods:The co-culture system of Human embryonic stem cells (H9) and liver cancer HepG2 cells was established.The effect of H9 on the biological behavior of HepG2 cells was observed by microscope,the flow cytometry was used to detcct the apoptosis of tumor cells and the cell cycle alteration.Transwell assay was used to detect the migration and invasion of tumor cells.Gene microarray technique was used to examine the change of gene expression profile of HepG2 cells.Results:In the process of co-culture,the growth of hepatoma cells was inhibited.With the extension of the culture time,cells decreased gradually,and occurred signs of aging or apoptosis.Flow cytometry test results showed that the apoptosis rate of hepatoma ceils was significantly increased,and the cell cycle was blocked in the G0/G1 phase.Transwell test results showed that the invasion and migration of HepG2 cells were decreased.The gene chip results showed that the whole genome expression profile of HepG2 cells had a significant change.Conclusion:The human embryonic stem cells had an inhibitory effect on HepG2 cells in vitro.

Objective To explore the association between sleep quality and the increasing risk of type 2 diabetes mellitus (T2DM).Methods A total of 771 patients aged 25-70 years living in Xuzhou City of Jiangsu Province for at least 5 years were enrolled for the survey of risk factor related noninfectious chronic disease in 2013.In this investigation,those who suffered from other types of diabetes,neuropathy,other endocrine disease,cardiovascular,renal and hepatic dysfunction,dyspnea or cancer were excluded.To reduce the influence of confounding factors,another 771 participants were enrolled as controls.Each case was arranged to have a control who was matched in age (difference not more than 3 years),gender,residence and family history.All the participants were interviewed with self-designed questionnaire,and sleep quality was measured by Pittsburgh Sleep Quality Index (PSQI) questionnaire.Student's t test,Chi-square and multivariate logistic regression were used for data analysis.Results The PSQI score in the T2DM patients vs.the controls were 5.15±2.40 vs.2.71 ± 1.93 (t=21.96,P＜0.01).The scores of sleep-related factors,including subjective poor sleep quality,bedtime resistance,short sleep duration,sleep efficiency,sleep disturbance,use of sleep medication and daytime dysfunction,of the T2DM patients were higher than those of the controls.The proportion of sleep related behaviors of the T2DM patients was higher,except for early awakening,cold feeling and nightmare.Poor sleep quality was associated with the increasing risk of T2DM (odds ratio 2.06,95％ CI 1.69-2.52).In multivariate logistic regression,when adjusted for confounding factors,the risk of T2DM was still increased (odds ratio 1.72,95％ CI 1.62-1.83).Sleep-related factors (e.g.subjective poor sleep quality,bedtime resistance,short sleep duration,sleep efficiency and sleep disturbance) were correlated with the risk of T2DM (odds ratio was 3.34,1.63,1.10,1.87 and 3.89,respectively).Conclusion Low quality of sleep may be strongly associated with an increased risk of T2DM.

HPLC analysis was performed to study the changes in chemical composition of ginseng extracts prepared from high quality ginseng with 0, 2, 4, 8 h of steamed times. An UFLC-MS/MS multiple-reaction monitoring (MRM) quantitative analysis was made to investigate the pharmacokinetic behavior differences of ginsenosides in mice ig administered of ginseng extracts with different steamed times in the negative ion mode, with Digoxin as the internal standard substance. The mice were injected with LPS to establish inflammation model after ig administration of ginseng for a week and the contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in mice plasma were detected by ELISA, in order to study on anti-inflammatory effects of ginseng with different steamed times. It was determined that levels of TNF-α and IL-1β were significantly decreased in inflammation model group ig administered of ginseng extracts with 8h of steamed time. The results showed that the chemical components in ginseng changed after steaming and the components into the blood changed, correspondingly. Ginseng with steamed 8 h contributes to anti-inflammatory effects. These results provided an experimental basis for revealing the active substance basis and dose-effect relationship of ginseng on anti-inflammatory effect.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacokinetics ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Ginsenosides ; chemistry ; pharmacokinetics ; Hot Temperature ; Humans ; Inflammation ; drug therapy ; Male ; Mice ; Mice, Inbred ICR ; Panax ; chemistry ; Time Factors

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