Object To establish a high efficient and reliable method for extraction and analysis of the diterpenoids in Pteris semipinnata L. Methods Supercritical CO 2 fluid modified with alcohol was used to extract the diterpenoids in P. semipinnata, the extracting conditions were optimized by orthogonal design method, and the modifying solvent was investigated through total ions chromatograghy normalization. A quadrupole mass spectrometer coupled with atmospheric pressure chemical ionization interface was employed as a detector for HPLC, the -1 ion was used as selective ion for the detecting of ent 11 ? hydroxy 15 oxo kaur 16 en 19 olic acid (5F) in selective ion monitoring (SIM) mode. The peak area of SIM and tolal ion chromatogram (TIC) were used for quantitative determination. As an example of its application, this method was used to determinate the content of 5F as antitumor diterpenoids in P. semipinnata. Results The optimized conditions for supercritical CO 2 fluid extraction are 25 MPa, 60 ℃, 15% methanol, flow rate 3 0 mL/min; analytical column was Diamonsil ODS (150 mm?4 6 mm, 5 ?m); the mobile phase of HPLC was CH 3CN 2 mmol/L NH 4AC (35∶65), flow rate 1 0 mL/min, injection volume 5 ?L; the standard curve showed good linearity over the range of 0 05-2 5 ?g; the limit of detection is 0 4 ng; the recovery is 97 8% (n=3). Conclusion This method is highly efficient, sensitive, and selective, which can be applied to study the antitumor drug of diterpenoids in P. semipinnata and to establish the drug standard.

OBJECTIVE:To prepare solid dispersion for increasing solubility of quercetin METHODS:Using polyvinyl pyrrolidone(PVP) as the carrier,solid dispersion of quercetin was prepared by solvent -volatilization method The solubility of the solid dispersion was detected IR spectrometer and UV spectrometer were used to investigate the physical and chemical properties of the solid dispersion RESULTS:The solubility of solid dispersion was significantly increased compared with quercetin and the physical mixture Quercetin existed as ultrafine crystalline or molecular form in solid dispersion,so there was no chemical reaction between quercetin and PVP CONCLUSION:PVP could be used as the carrier of solid dispersion to increase the solubility of quercetin in water

OBJECTIVE:To establish a method for determination of the content of quercetin in quercetin-PVP solid dis?persions.METHODS:The detection wavelength for dertermination of quercetin content was set at374nm.RESULTS:The caliberation curve was linear in the range of5.0～25.0?g/ml(r=0.9998).The average recovery was99.12%,RSD=0.70%(n=6).CONCLUSION:This method is stable,quick,accurate and simple,and is suitable for quality control of quercetin in quercetin-PVP solid dispersions.

Objective To approach the effect of cathepsin K(CK) on the onset of osteoporosis,and to study the influence of Bushen Tongluo Prescription(BTP,with the actions of invigorating the kidney and dredging the meridians) on osteoclast CK in ovariectomized rats.Methods Thirty female SD rats were divided into three groups randomly: sham-operation group,model group and BTP(12.46g.kg-1.d-1) group.The bilateral ovaries in rats of the model group and BTP group were removed.Four weeks later,BTP group were given gastric gavage of BTP.On the 6th and 12th weeks,we detected the general bone density of rats,examined CK expressionin osteoclasts with Western blot and reverse transcription-polymerase chain reaction(RT-PCR) methods.ResultsThe in-vitro cultured cells had the phenotype feature of mature osteoclasts.The bone mineral density of ovariectomized rats was negatively correlated with CK expression in time-effect manner.BTP down-regulated the CK protein and CK mRNA expression(P

Objective:To investigate the role of thoracic echocardiography in diagnosis and differential diagnosis of the right ventricular outflow tract into the right aorta with right ventricular aneurysm rupture.Methods:A total of 19 patients with sinus rupture caused by ventricular septal defect in the right ventricular outflow road were reviewed.These patients were diagnosed as aortic right coronary sinus by preoperative transthoracic echocardiography from February 2010 to September 2016 in Xiangya Second Hospital and Xiangya Medical College Affiliated Haikou Hospital.The clinical symptoms,echocardiography and intraoperative lesions were compared and analyzed among the patients.Results:The intraoperative lesions were consistent with the preoperative echocardiographic diagnosis in 16 patients.Among them,11 cases could be heard the continuous sound of the machine with systolic murmur and no tremor in left sternal rib 2-3 auscultation.The color Doppler imaging of right ventricular systolic indicated the based continuous Doppler spectrum.The preoperative echocardiography and intraoperative lesions testified the ventricular septal defect size and the right ventricular outflow tract into the right aorta with right ventricular aneurysm rupture.Among them,3 cases were complicated with other heart diseases.Other 3 cases were not accorded with the preoperative diagnosis.There were auscultation smell and noise but no tremor.The preoperative echocardiography and lesions indicated the right ruptured aneurysm of the sinus of big and long mouth,1 case with aortic valve stenosis and regurgitation,congenital aortic valve two valve malformation,1 case with aortic valve-severe reflux,1 case with pulmonary infection,and 3 cases with no intraoperative lesions in the ventricular septal defect.Conclusion:Preoperative transthoracic echocardiography is the preferred preoperative,quick and noninvasive method for ruptured aneurysm.Each stage of observation procedure and method can observe the ruptured aneurysm of aortic sinus,the location,size and adjacent ofventricular septal defect.Meanwhile,the flow spectrum characteristics of the ventricular septal defect in the right outflow tract of the right sinus aneurysm can be found by Doppler ultrasound imaging.

Objective:To investigate the exression of miR-34a on lung cancer and normal lung tissues,and the effect and mechanism of miR-34a in lung cancer cell invasion and migration.Methods: qPCR was used to detect the expression of miR-34a on lung cancer.miR-34a-mimic and miR-34a-inhibitor were used to overexpress and knockdown miR-34a.qPCR was used to detect the effectiveness.Western blot was used to detect the expression of Snail after induced with miR-34a-mimic and miR-34a-inhibitor.Luciferase reporter gene was used to detect interaction between miR-34a and Snail.Transwell invasion assay was used to detect invasion ability after induced with miR-34a-mimic and miR-34a-inhibitor.Scratch assay was used to detect migration ability after induced with miR-34a-mimic and miR-34a-inhibitor.The expression of E-cadherin,Vimentin and Twist were detected by Western blot.Results: miR-34a expression was significantly reduced in lung cancer.With the stage of lung cancer progression,the expression of miR-34a reduced.With the differentiation of lung cancer progression,the expression of miR-34a decreased.Decreasing of miR-34a was associated with lung cancer lymph node metastasis.miR-34a-mimic and miR-34a-inhibitor could overexpress and knockdown miR-34a.miR-34a could regulate expression of Snail.Snail was the direct target of miR-34a;miR-34a could regulate the invasion ability of human lung carcinoma H1650 cells;miR-34a could regulate the migration of human lung carcinoma H1650 cells;miR-34a could regulate the expression of E-cadherin,Vimentin and Twist.Conclusion: miR-34a plays the role of tumor suppressor factor in lung cancer.miR-34a can regulate the invasion and migration ability of lung carcinoma H1650 cells by Snail induced EMT.

Objective:To investigate the inhibitory effect of perillyl alcohol (PA) on the proliferation and invasion of tung cancer cell A549,and the influence of PA on tumor angiogenesis was studied.Methods:Different concentrations of PA and erlotinib were added into lung cancer cell A549,the inhibiting effect of drug group on lung cancer cell A549 was found by MTT assay.The inhibiting effect of PA on lung cancer cell A549 invasion was measured by Transwell assay.ROS changes of PA on lung cancer cell A549 was detected by fluorescent.Influence of PA on Caspase-3 activity of lung cancer cell A549 was measured by spectrophotometry,VEGF,HIF-1 α,COX-2 expression in lung cancer cell A549 was measured by Western blot,and the NF-κB activity of lung cancer cell A549 was measured by EMSA.Results:Compared with blank control group,cell growth inhibition rate of PA and erlotinib on lung cancer cell A549 was increasing with the increased concentrations (10,50,100 μ,g/ml),the difference was statistically significant (P＜ 0.05),the invasion ability of lung cancer cell A549 was decreased continuously,the difference was statistically significant (P＜0.05).The ROS level of lung cancer cell A549 had no obvious change with the increasing density of erlotinib,but obviously increased with the increasing concentrations of PA (10,50,100 μg/ml).With the increasing concentrations of PA,the expression of COX-2,VEGF and HIF-1α were continuously decreased.EMSA assay showed that NF-κB was continuously decreased with the increasing concentrations of PA.Conclusion:The antitumor mechanism of PA on lung cancer cell A549 might be related to increase the expression level of ROS and reduce the expression of activity of NF-κB,COX-2,VEGF and HIF-1α with angiogenesis signaling pathway.

To compare the effects of laparoscopic total or subtotal hysterectomy and myomectomy on ovarian reserve by measuring the serum level of anti-Mullerian hormone (AMH).A total of 96 patients with uterine fibroids underwent total laparoscopic hysterectomy (TLH group,n =32),laparoscopic subtotal hysterectomy (LSH group,n =31) and laparoscopic myomectomy (LM group,n =33).Changes in ovarian reserve were examined by measuring the level of AMH pre-operation and at 1 and 4 months post-operation.No significant differences existed in AMH in LM group[(1.42 ±0.65),(1.31 ±0.53) & (1.33 ±0.61) μg/L,P ＞ 0.05].The levels of AMH in both groups were significantly lower than those at pre-operation [(1.17 ± 0.11),(1.01 ±0.10),(0.48±0.54) & (1.18±0.93),(0.45 ±0.39),(0.14±0.0) μg/L,P＜0.001].The mean percentage decrease in AMH was lower in TLH group than that in LSH group at 4-month follow-up [(-37.4 ± 37.6) ％ vs.(-27.0 ± 23.0) ％,P =0.017].The results showed that LM had no effect on ovarian reserve.Both LSH and TLH had adverse effects on ovarian reserve and TLH was more marked.

Aim To study the effect of 5F on tubulin polymerization.Methed 5F was added into the tubulin polymerization-depolymerization system and the changes of Absorption values at 350 nm, 37 ℃ were recorded. Results 5F decreased the Absorption values of the reaction system.and markedly inhibited not only tubulin polymerization but also tubulin depolymerization. Conclusion 5F inhibits tubulin polymerization and depolymerization, and leads to cell cycle arrested, and possibly induces apoptosis.

AIM To find a new method for purifying the active compound 5F isolated from Pteris semipinnata L.(PsL ) and observe its enhanced cytotoxicity when combined with other antitumor agents. METHODS Silica gel combined with AgNO 3 was made to purify 5F. Cytotoxicity was detected with trypan blue dye exclusion assay. RESULTS After purification, the concentration of compound 5F in purified products was higher than 99%. The inhibitory rates of 5F combined with 5 Fu or CDDP or VCR were higher than that of these drugs used alone. CONCLUSIONS Compound 5F could be purified effectively using silica gel combined with AgNO 3. 5F could enhance the cytotoxicity on HL 60 and K562 cells of the drugs mentioned above. The different effect of these agents on the various phases of cell cycle kinetics may explain the enhanced effect of 5F.