Objective To optimize the flash type extraction ofDanzhuye Granules.Methods With the total flavonoids and extract yield as indices, the orthogonal test was adopted to optimize the extraction times, the ethanol concentration, solid-liquid ratio, and the duration of extraction forDanzhuye Granules.Results The factors that infuenced the extract efficiency from high to low were the extraction times, volume fraction of ethanol, the ratio of liquid to solid, and the extraction time. The optimum extracting condition included that the origin was extrated by the ethanol volume fraction 80% with the material to liquid ratio 30:1, total of 3 times, with each time of 3 minutes. Validation experiments of the potimal condition showed that the extraction of total flavonoids was 29.1 mg/g, and the extract yield was 23.6%.Conclusions The flash type extraction method is suitable for extraction, which provides a new method for the development and utilization ofDanzhuye Granules.

BACKGROUND:Silk fibroin/mesoporous glass ceramic composites have been reported to exert satisfactory repair outcomes in bone defects and hold good biocompatibility. However, the biosafety and preparation methods are rarely reported. OBJECTIVE:To investigate the preparation method and treatment outcomes of silk fibroin/mesoporous bioactive glass ceramic in skul repair. METHODS:Thirty-two Sprague-Dawley rats were enrol ed to establish the skul defect models and were thenrandomized into two groups:fibroin/mesoporous glass ceramic materials and silk fibroin were respectively implanted into the defect region in experimental and control groups. At 4 and 8 weeks after implantation, the CT examination and histological observation were performed. RESULTS AND CONCLUSION:CT examination showed that at 4 weeks after implantation, the defect area in the experimental group diminished in size, showing more dense new bones. The defect area of the control group was reduced, and a smal amount of new bones were observed. At 8 weeks after implantation, bone defect repair was completed in the experimental group, but not in the control group. The bone volume in the experimental group was significantly larger than that in the control group at different time points after implantation (P<0.05). Hematoxylin-eosin staining found that at 4 weeks after implantation, in the experimental group, there was new bone between the implant and the bone, which did not cause inflammation;there were few new bones and fibrous tissues in the control group. At 8 weeks after implantation, many new bones formed in the experimental group, with similar morphology to the host bone and the scaffold was degraded completely. Conversely, the implant material stil existed in the control group. In conclusion, the silk fibroin/mesoporous glass ceramic composite can promote bone repair.

Objective To study the biocompatibility of bone marrow mesenchymal stem cells (BMSCs) on acellular collagen matrix double-layer material.Methods BMSCs with acellular collagen matrix double-layer material were trained as experimental group,while separately cultured BMSCs as control group.The growth and proliferation of BMSCs on acellular collagen matrix double-layer material were investigated.Scanning electron microscope was used to observe the adherence of cells,and cell vitality was detected by CCK-8 method.Results The acellular collagen matrix double-layer material did not influence the growth and proliferation of BMSCs.The difference between the growth curves of experimental group and control group was not significant (P＞0.05).Conclusions The acellular collagen matrix double-layer material has acceptable biocompatibility.This research provides a scientific basis for the application of this composite material.

In the present study ,we aimed to observe the effect of curcumin on TSST-1-induced inflammatory cytokines in splenocytes of mouse and provide evidence for the further study on the effect of curcumin on inflammatory shock .Lactate de-hydrogenase (LDH) release assay was used to determine cytotoxicity of different doses of TSST-1 and curcumin .Inflammatory cytokines were determined by ELISA and flow cytometry .The doses of TSST-1 and curcumin we used in the present study did not cause significant cytotoxicity .TSST-1 induced higher level of IFN-γand IL-2 production but relatively lower level of TNF-α.The production of IL-1β,IL-6 and IL-12 was undetectable .TSST-1 induced Th1 cytokines (IFN-γand IL-2) were not IL-12-dependent which was different from LPS-induced IFN-γ.Curcumin significantly reduced IFN-γand TNF-αproduction at the concentration of 15 umol/L (P<0 .05) ,but had no effect on IL-2 production (P>0 .05) .It’s suggested that curcumin could significantly inhibit the production of IFN-γand TNF-αby splenocytes induced by TSST-1 ,but could not affect the prolifera-tion of T cells .

Objective To investigate the inhibitory effects of docosahexaenoic acid (DHA)and 5-fluorouracil (5-FU)in combination on human gastric cancer cell line AGS in vitro .Methods Human gastric cancer line AGS was treated with different concentrations of DHA and 5-FU alone or in combination.The inhibition of cell proliferation was evaluated by MTT assay.Dose of median (IC50 )of drugs (alone or in combination)and the combination index (CI)were calculated using the median-effect equation and the combination index equation of Chou-Talalay.Flow cytometry was used to detect the cell cycle distribution.The expression of mitochondrial respiratory membrane protein complex in AGS cells was analyzed with Western blot.Results DHA and 5-FU alone or in combination could markedly suppress the proliferation of AGS in significantly time-dependent and dose-dependent manners (P <0.05).IC50 values with DHA or 5-FU administered for 24 h and 48 h were 5 1.60 μg/mL (DHA:24 h),34.82 μg/mL (DHA:48 h),45.90 μg/mL (5-FU:24 h),and 1 6.86 μg/mL (5-FU:48 h), respectively.DHA remarkably strengthened the inhibitory effect of 5-FU and decreased IC50 of 5-FU by 3.56 -2.1 5 folds.The combination of DHA and 5-FU showed synergism.Flow cytometry showed that AGS cells treated with DHA and 5-FU were arrested in G0/G1 phase and the proportion of AGS cells in G0/G1 phase increased compared with that in the control group,DHA group and 5-FU group,while the proportion of the cells in S phase decreased significantly (P < 0.05 ).Western blot showed after treatment with DHA and 5-FU for 48 h,the expression of mitochondrial respiratory membrane protein complex was significantly decreased compared with control group,DHA group and 5-FU group (P <0.05).Conclusion DHA could act synergistically with 5-FU in inhibiting the growth of gastric carcinoma cells,and meanwhile decrease the dose of 5-FU.The mechanism may be associated with cell cycle arrest in G0/G1 phase and interference in the energy metabolism of AGS cells due to inhibition of the expression of mitochondrial oxidative respiratory chain complexes by the two compounds.

OBJECTIVE:To establish a method for determining the concentration of vinorelbine bitartrate in human plasma, and study the pharmacokinetic process of vinorelbine in human plasma. METHODS:The samples were extracted with Solid-phase extraction(SPE). LC-MS/MS was performed on the column of Agilent Extend C18 with the mobile phase of 5 mmol/L ammonium ac-etate solution(pH10.5)-acetonitrile-methano1(18∶10∶72,V/V/V)at the flow rate of 0.4 ml/min,the ion source was ESI and MRM mode was used to scan positive ion detection. The ion reaction of quantitative analysis was m/z 779.50→m/z 122.10 (vinorelbine) and m/z 811.60→m/z 224.50 (internal standard, vinblastine). 3p97 was used to calculate the pharmacokinetic parameters. RE-SULTS:The determination was not interfered by endogenous impurities,and there was no matrix effect;the lowest limit of quantita-tive analysis was 2.0 ng/ml with the linear range of 2.0-4 000.0 ng/ml(r=0.997 8);the linear range of vinorelbine in plasma was. The intra-day and inter-day precisions and accuracy results were all in line with the acceptable limit across all concentrations. t1/2γ of 30 mg/m2,40 mg/m2 Vinorelbine Emulsion for Injection groups and 30 mg/m2 Vinorelbine for Injection group were(37.958 ± 34.256)、(47.835±54.231)、(76.873±40.537)h respectively,cmax were(1 426.250±397.562)、(1 700.125±624.669)、(2 187.500± 828.040)ng/ml respectively,AUC0-48 h were(75 839±19 551)、(82 088±14 207)、(95 318±18 208)mg·h/L respectively. CONCLU-SIONS:The method is rapid,sensitive and specific,and suitable for the determination of vinorelbine and pharmacokinetic study. Metabolic processes of vinorelbine can be described as first order process of three-compartment model in cancer patients.

Objective To discuss the clinical application of multi-slice spiral CT angiography (MSCTA) in acute spontaneous intracranial hemorrhage .Methods 41 cases of acute spontaneous intracranial hemorrhage diagnosed with CT (including subarachniod cavity hemorrhage 29, intra-cerebral hematomas 12)were examined with MSCTA,the techniques of volume rendring (VR) and maximum intensity projection(MIP)were used. 7 cases of intracranial aneurysm closed by titanic clamp , 2 cases of cerebral arteriovenous malformation(AVM) and one case of AVM treated by intra-zest aneurysm excision,the post-operative MSCTA examination were received.Results 11 cases of intra-cerebral aneurysm, 4 cases of cerebral AVM and 1 case of AVM with intra-zest aneurysm were found among the 41 cases of acute spontaneous intracranial hemorrhage. MSCTA clearly demonstrated the size,neck and artery of aneurysms as well as the location, size ,zest shape, artery and vein of AVM. Pre-operative MSCTA findings of 7 cases of intracranial aneurysm clapped by titanic clap ,2 cases of cerebral AVM and 1 case of AVM treated by intra-zest aneurysm excision were confirmed with the operative findings.Post-operative MSCTA showed that the clamp location was normal; arteries were passable; zests of AVM were excised. Conclusion MSCTA is a non-injured, rapid and effective way to find the cause of acute spontaneous intracranial hemorrhage.It also has clinical application of post-operatiove assessment for intracranial aneurysm and cerebral AVM.

Objective:To study the technical operation of self-developed investment (FUS-invest) for titanium crown and bridge. Methods:Orthogonal design was done. Four factors were taken into account, i.e. water/powder ratio, adhesive/hardener ratio, fine powder proportion in refractory and agitation time. And each factor was ranked into three levels. Slurry fluidity, initial setting time and setting time of every experiment group were studied. Grain size graduation of the best group was measured.Results:Fluidity, initial setting time and setting time were most greatly affected by water/powder ratio and secondly by fine powder proportion in refractory. When water/powder ratio was 7.5∶1 and the content of fine powder was 35%, slurry fluidity was better and initial setting time and setting time were appropriate. When the content of fine powder of AFS fineness number 300 was 31.31%, fineness number 100 was (40.09%), fineness number 70 was 18.05% and adhesive/hardness ratio was 200∶1, the perfect titanium castings could be fabricated. Conclution:The best rules of technical operation for perfect titanium castings are water/powder ratio (7.5)∶1, the content of fine powder 35% and adhesive/hardener ratio 200∶1.

Objective To construct the eukaryotic expression vector of human fibrinolysin for further study on anti-thrombus and thrombolysis.Methods Human fibrinolysin gene was amplified from human fetal liver tissue with PCR,and was cloned into eukaryotic expression plasmid pCI-neo and then sequenced.After the correct identification by sequencing,the eukaryotic expression recombinant plasmid of human fibrinolysin was constructed and identified with PCR and enzyme digestion.Results The PCR amplification product of fibrinolysin gene was 1 740 bp.The result of sequencing was the same as that registered in GenBank.The results of PCR and enzyme digestion showed that the recombinant eukaryotic expression plasmid had correct codogenic gene fragment.Conclusion The eukaryotic expression recombinant plasmid of human fibrinolysin gene is successfully constructed and identified.

Objective To investigate and summarize the MRI characteristics of splenic sclerosing angiomatoid nodular transformation (SANT).Methods A retrospective analysis of 5 SANT cases were analyzed,in terms of their MRI characteristics and pathological findings.Results MRI findings of SANT included:T1WI presents iso-signal or slightly low signal,all displayed lesions were detected as low signal compared with spleen,but higher than muscle signal on T2 WI,and with speck dots or starlike low signal in the central area,without necrosis and cystic change.The signal was significantly differentiated compared with the spleen on DWI.On chemical shift imaging,2 cases were showed slightly higher signal on out phase,the others without signal change.On enhanced scan,4 cases had edge obvious enhancement on arterial phase,inward filling enhancement,and the signal was higher than the spleen,1 case without arterial phase enhancement,but with mild concentric delayed enhancement.All of the speck dots and starlike areas decreased with time delay,with certain degree enhancement on delayed phase.Conclusions There were some MRI features of SANT,preoperative MRI can prompt diagnosis,but final diagnosis depends on pathology.