Objective To establish a method for the determination of Chrysophanol in Jiangzhiling Capsules. Methods RP-HPLC was used for samples separating on a Nova-Pak C18 column. The mobile phase consisted of methanol-0.1 %H3PO4(85 ∶15). The detection wavelength was 254 nm and the flow rate was 1 mL?min-1.Results The standard curve of Jiangzhiling Capsules was linear in the rang of 0.037 5～0.750 0 ?g (r=0.999 9) with average recovery of 98.31 %(RSD=1.36 %). Conclusion The method is simple,accurate and precise. It can be used effectively for the quality control of Jiangzhiling Capsules.

Objective To investigate the effect of sodium valproate in suppression of cell proliferation and arrest of cell cycle of in human hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988.Methods Hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988 were inoculated on the culture plate,cultured in the DMEM medium,they were intervened with sodium valproate in concentration of 0.2 mmol/L (0.2 mmol/L group),1.0 mmol/L (1.0 mmol/L group),5.0 mmol/L (5.0 mmol/L group) and dimethyl sulfoxide (control group) for 48 h respectively.Absorbance was measured by enzymelinked immunosorbentassay equipment,and inhibition ratio was calculated.Cell cycle was detected by flow cytometry.Results The absorbance of hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988 in 5.0 mmol/L group were significantly lower than those in control group and 0.2 mmol/L group (0.569 ±0.059 vs.0.706 ±0.033 and 0.760 ±0.020,2.068 ±0.178 vs.2.793 ±0.144 and 2.663 ± 0.078,P ＜ 0.05),the absorbance of pancreatic cancer cell line PaTu8988 in 1.0 mmol/L group (2.432 ± 0.084) was significantly lower than that in control group and 0.2 mmol/L group (P ＜ 0.05).With the sodium valproate concentration increased,inhibition rate of tumor cell increased gradually,the inhibition rate of hepatoma cell line SMMOL/LC-7721 and pancreatic cancer cell line PaTu8988 in 5.0 mmol/L group was 23.5％ and 25.9％ respectively.Compared with control group,with the sodium valproate concentration increased in 0.2,1.0,5.0 mmol/L group,the proportion of G1 phase cell increased gradually in hepatoma cell line SMMOL/LC-7721 [(49.25 ± 1.63)％,(65.26 ± 2.34)％,(83.13 ± 1.78)％ vs.(49.22 ± 4.35)％],the proportion of S phase cell decreased gradually [(26.84 ± 2.30)％,(17.76 ± 3.90)％,(3.38 ± 0.65)％ vs.(29.21 ± 2.35)％],cell cycle showed G1 phase arrest,there was significant difference (P ＜ 0.05).Compared with control group and 0.2 mmol/L group,the proportion of G2 phase cell increased in pancreatic cancer cell line PaTu8988 in 1.0 and 5.0 mmol/L group [(26.80 ± 1.50)％,(36.58 ± 3.78)％ vs.(12.00 ± 4.62)％,(16.54 ± 2.26)％],cell cycleshowed G2 phase arrest,there was significant difference (P ＜ 0.05).Conclusion Sodium valproate mightsignificantly suppress the cell proliferation in hepatoma cell line SMMOL/LC-7721 and pancreatic cancercell line PaTu8988 and induce cell cycle arrest,it is clinically promising antitumor drug.

Objective To construct the recombinant plasmid pCMV-Myc-PIASX? and express the fusion protein in mammalian cells.Methods PIASx? fragment was digested from the original vector pGADT7 with SalⅠand NotⅠ,and then was inserted into the targeted pCMV-Myc vector by the recombinant DNA technique.After identification,the recombinant plasmid was transfected into CHO cells.The expression of recombinant Myc-PIASx? fusion protein was detected by Western blotting.Results By the restriction enzyme digestion,fragment purification,ligation and transformation,the recombinant plasmid was obtained.The right recombinant plasmid pCMV-Myc-PIAS3 was identified with enzyme digestion and sequencing.By EcoRⅠ digestion analysis,pCMV-Myc-PIASx? showed a 5641 bp band.By XbaⅠdigestion analysis,pCMV-Myc-PIASx? showed two expected band of 3291 bp and 2349 bp.A specific protein expression band at 68 000(PIASx? fusion protein) was showed in Western blotting,which matched recombinant plasmid.Conclusion The recombinant plasmid of pCMV-Myc-PIASx? is sucssesefully constructed,which provids a good tool for further function study on PIAS family.

Objective To investigate the medication rules for the treatment of hepatopathy by national medical masters. Methods The prescriptions for hepatopathy were collected to build a database based on Traditional Chinese Medicine (TCM) inheritance assist system. After analyzed by the data mining, such as apriori algorithm and complex system entropy cluster, the frequency of single medicine and drug combinations that used in prescriptions, and the association rules among drugs were obtained. Results The most frequently used drugs were Herba Artemisiae Capillaris, Tuckahoe, Radix Salviae Miltiorrhizae, Rhizoma Atractylodis Macrocephalae, and Radix Bupleuri, et al. The most frequently used drug combinations were “Radix Salviae Miltiorrhizae, Tuckahoe”, “Rhizoma Atractylodis Macrocephalae, Tuckahoe”, and “Radix Salviae Miltiorrhizae, Herba Artemisiae Capillaris”, et al. And when the confidence of drug association rules was 1, the drug combinations were“Radix et Rhizoma Rhei, Giant Knotweed→Herba Artemisiae Capillaris”,“Radix et Rhizoma Rhei, Radix curcumae→Herba Artemisiae Capillaris”, “Giant Knotweed, Gardenia→Herba Artemisiae Capillaris” and so on. Conclusion The medicines were often with the actions of clearing heat, removing dampness, regulating qi and activating blood circulation, which were used for the treatment of hepatopathy by national medical masters.

Objective To study the effects of Weibimei and its Chinese medicine portions(cortex frangulae, fennel powder, acorus tatarinowii, glycyrrhizae and asparagus) on cell cycle regulation in gastric cancer cells and growth inhibition in gastric cancer Xenografts.Methods Gastric cancer SGC-7901 cells were normally cultured and treated with Weibimei, extractum glycyrrhizae, cortex frangulae, fennel powder, acorus tatarinowii and asparagus.Then, the proliferation of SGC-7901 cells were detected by MTT assay, their effect on SGC-7901 cell cycle were measured by flow cytometry, and the expression of cell cycle related proteins were examined by Western blot.Animal models of SGC-7901 cells xenografts in nude mice were constructed to evaluate the growth inhibition effects of Weibimei and its Chinese medicine portions, and the expression of STAT3,p-STAT3, Cyclin D1, Bcl-2 and Survivin in gastric tumors were detected by immunohistochemical assays.Results Weibimei and its three kinds of Chinese medicine portions, cortex frangulae, fennel and acorus tatarinowii could obviously inhibit the proliferation of SGC-7901 cells, compared to control group, with a statistical significant difference (P<0.05).The cell cycle of SGC-7901 was arrested at G1 phase with cortex frangulae, fennel and acorus tatarinowii treatment, the expression of Cyclin A, B, D, E were decreased significantly (P<0.05).However, no significant effects was found in glycyrrhizae or asparagus group.Xenografts tumor sizes in Weibimei, cortex frangulae, fennel or acorus tatarinowi groups were much smaller than that in saline group or Triple Therapy group (three-kinds-drugs-combination for the treatment of gastric ulcer) (P<0.05).Xenografts experiments showed that the tumor growth in nude mice in Weibimei, cortex frangulae, fennel or acorus tatarinowi groups were significantly smaller than that in control group or in Triple Therapy Group (P<0.05), and phosphorylated STAT3, and STAT3 signaling targeted genes, including Bcl-2, Cyclin D1 and Survivin were all significantly down-regulated by immunostaining.Conclusion Weibimei and its Chinese medicine portions including cortex frangulae, fennel and acorus tatarinowii can could significantly inhibit the growth of SGC-7901 cells proliferation and arrested the cell cycle of G1 phase in vitro, and inhibit gastric cancer xenografts in nude mice in vivo.

Objective:To apply modified proximal femoral nail anti-rotation ( PFNA ) fixation tech-niques performed by percutaneous placement of guide wire combined with true lateral view and to make the procedures simpler.Methods:A retrospective study was used to analyze the clinical data of femoral intertrochanteric fractures cases, which were treated with conventional PFNA fixation or modified PFNA fixation performed by percutaneous placement guide wire combined with true lateral view in our hospital, from March, 2011 to May, 2014.In the study, 60 cases were followed for average 13 months.The oper-ation time, the amount of bleeding, the fluoroscopy time, postoperative radiographic measurements ( tip apex distance, TAD ) and hip function scores were analyzed.Results:In modified PFNA group, the amount of bleeding, the operation time and the fluoroscopy time were (70.5 ±12.5) min, (34.9 ±6.1) mL, ( 63.6 ±9.7 ) s respectively.In conventional PFNA group, they were ( 80.6 ±17.1 ) min, (47.8 ±6.7) mL, (68.5 ±8.7) s respectively.There were significant differences in the above respects between the two groups (P were 0.006, 0.013, and 0.022 respectively).There were no significant differences in TAD, fracture healing time, postoperative hip scores between the two groups (P>0.05). Conclusion:Fracture line is a natural entry point for some cases of femoral intertrochanteric fractures when we use proximal femoral nail anti-rotation to fix the fracture.Applying percutaneous insertion of the guide pin combined with true lateral view could reduce the operation time, amount of bleeding, and fluo-roscopy time significantly, make the procedures simpler and acquire satisfactory results .

Objective To explore the ideal treatment mode of brain metastases by Meta-analysis.Methods Articles were searched for from the databases at home and abroad using English and Chinese keywords.Searching time limited from the databases setting up to December 30,2012.Jadad score was applied to evaluate the qualities of literatures.RevMan5.0 software was applied to perform the Meta-analysis.A totle of 25 articles including 2 750 patients were eligible for the Meta-analysis,which divided into groups with different treatment.Results Compared with monotherapy,combined therapy improved 1-year survival (OR =0.58,95％ CI:0.46 ～0.71,P ＜0.000 01).In combined therapy groups,compared with two methods,three kinds of therapies improved 1-year survival (OR =0.63,95 ％ CI:0.50 ～ 0.80,P =0.000 1).Compared with local therapy only or systemic therapy only,systemic combined local therapy improved 1-year survival (OR =0.68,95％ CI:0.53 ～ 0.86,P =0.001 ; OR =0.59,95％ CI:0.41 ～ 0.86,P =0.006).In systemic combined local therapy groups,three kinds of treatments improved 1-year survival compared with two methods (OR =0.52,95％ CI:0.35 ～ 0.78,P =0.002).Compared with non-molecular targeted therapy,molecular targeted therapy improved 1-year survival (OR =0.76,95％ CI:0.67 ～ 0.87,P ＜ 0.000 1).Conclusion The reasonable treatment for patients with brain metastases is combined treatment with operation,radiotherapy and chemotherapy.There is better curative effect added molecular targeted therapy based on original scheme,if patients have targeted therapy indications.

Objective To construct the eukaryotic expression recombinant PIAS3 plasmid fused with Myc protein and express the fusion protein Myc-PIAS3.Methods The full length PIAS3 fragment of 1 851bp was amplified by PCR and ligated into pMD18-T vector. The full length PIAS3 fragment was subcloned into eukaryotic pCMV-Myc vector between SalⅠ and NotⅠ sites.The recombinant pCMV-Myc-PIAS3 plasmid was trandfected into PC3 cells.The eukaryotic expression Myc-PIAS3 protein was checked by Western blotting with Myc antibody.Results The recombinant plasmid showed right sequence by the full length sequencing.The recombinant plasmid of pCMV-Myc-PIAS3 was identified by enzyme digestion.As expected,by EcoRⅠ digestion,it showed two bands of 4 357bp and 1 318 bp. By XbaⅠdigestion,it showed two bands of 3 291 bp and 2 384 bp.The sequencing result showed a N-terminal Myc of 13 amino acids followed with PIAS3 gene sequence in right reading frame.The pCMV-Myc-PIAS3 plasmid was transfected into prostate cancer PC3 cells.A specific protein expression band at relative molecular mass 68 000 was obtained by using Myc-antibody with Western blotting method.Conclusion The recombinant plasmid of pCMV-Myc-PIAS3 is sucssesefully constructed,and Myc-PIAS3 fusion protein is sucssesefully expressed.

Objective To investigate the effects of valproic acid ( VPA) on cell proliferation and cell cycle in human pancreatic cancer cell line PaTu8988 in vitro. Methods PaTu8988 cells were treated with VPA in concentration of 0, 0.2, 1.0 or 5.0 mmol/L for 24 h and 48 h respectively. Cell viability was measured by WST-8 assay. Cell cycles were detected by flow cytometery. Dimethyl sulfoxide added to the medium was used as blank control group, while PBS added to the medium was used as PBS group. Results After VPA treatment for 24 h, the inhibition rate of VPA 5.0 mmol/L group was 18.9% , which was significantly higher than those in control group, PBS group and VPA 0.2, 1.0 mmol/L group (0, 4.4% , 6.8%, 6.1% , P <0.05). After 48 h, the inhibition rates of VPA 1.0, 5.0 mmol/L were 12.9%, 25.9% , which was significantly higher than those in control group, PBS group and VPA 0.2 mmol/L group (0, 6.2% , 4.6% , P <0.01). After VPA treatment for 24 h, the proportions of G2 phase cell in VPA 1.0, 5.0 mmol/L group were ( 26.57 ± 1.88) % , ( 34.11 ± 4.74 ) % , which was significantly higher than those in PBS group, control group, VPA 0.2 mmol/L group [(10.72 ± 2.02)% , ( 13.53 ± 2.28)% , (13.81 ±2.40)%, P <0.01 ], the changes 48 h after VPA treatment was consistent with the changes 24 h after VPA treatment. Conclusions VPA may significantly suppress the cell proliferation of human pancreatic cancer cell line PaTu8988, and induce cell cycle arrest in G2 phase in a time and dose-dependent manner.

Objective To perform the ligand-based computer-aided drug design and construct the pharmacophore model of(1,3)-β-D-Glucan Synthase(GS)small molecule inhibitors.Method Six small molecules with diverse structures and good inhibitory activity were selected to construct the training set.The HipHop algorithm in Catalyst pharmacophore generation module was utilized to construct the pharmacophore models.The pharmacophore models were evaluated by constructed Decoy-set 3D database.Results Pharmacophore 02 has a good enrichment factor,sensitivity and specificity parameters.Pharmacoph-ore model validation with Decoyset 3D database proved that the model has good distinguishing capability.Conclusion The pharmacophore model of GS small molecule inhibitors was constructed and tested.It will provide valuable information for de-sign and discovery of novel small molecule GS inhibitors.