ABSTRACT:Objective To study the effects of propofol on the metastasis of tumor cells related PI3K/Akt signaling pathway.Methods The breast cancer model was established by transplanting human derived breast cancer cell lines into immunodeficient mice with naked gene.The mice,inoculated successfully,were randomly divided into 4 groups:control group (C group,n =6),propofol group (P group,n =6),propofol+PI3K inhibitor (BYL71 9)group (P+B group,n =6),and PI3K inhibitor group (BYL71 9)(B group,n =6).The expressions of PI3K,p-Akt and Akt were examined by Western blot at week 4 after administration;the gene levels of PI3KR1, Akt1 and Akt2 were detected by RT-PCR at week 4 after administration;the number of metastatic lung nodules from both lungs was also observed at week 4 after administration.Results Compared with those in C group,the expressions of PI3K and p-Akt were significantly higher in P group (P <0.05),the level of PI3KR1 mRNA but not Akt1 and Akt2 mRNA was significantly increased(P < 0.05 ),and metastatic lung nodules significantly decreased (P <0.05).In B group,the expressions of PI3K and p-Akt were significantly decreased (P <0.05 ),the levels of PI3KR1,Akt1 and Akt2 mRNA were not significantly increased (P >0.05),but metastatic lung nodules significantly increased (P < 0.05 ).Compared with those in B group,in P+ B group the expressions of PI3K and p-Akt were markedly higher (P <0.05),the level of PI3KR1 mRNA but not Akt1 and Akt2 mRNA was significantly increased (P <0.05),and metastatic lung nodules significantly decreased (P <0.05).Conclusion Propofol can inhibit the metastasis of tumor cells through the upregulated and activated PI3K/Akt signaling pathway.

Objective To analyze the image characteristics of chest schwannoma in coincidence 18FFDG SPECT studies,and to evaluate its prospective diagnostic function on chest schwannoma.Methods Four cases confirmed as schwannoma by surgery and pathology were enrolled in this retrospective study.Enhanced CT and coincidence 18F-FDG SPECT studies were performed before surgery.Imaging characteristics for the diagnosis of chest schwannoma were summarized.Results Tumors were totally removed by surgery.The histological diagnosis of schwannoma was confirmed by pathological,HE and immunohistological staining examination (positivity for the S-100 protein and vimentin),including 1 case of malignant schwannoma and 3 cases of benign.Four cases all showed high uptake of 18F-FDG.The uptake in malignant schwannoma was much higher than that in benign schwannoma.Benign schwannomas were noninvasive masses and mainly showed compression symptoms,with obvious lobulation and distinct borders.Malignant schwannoma was invasive masses.Conclusions 18 F-FDG coincidence SPECT is of limited value as a prospective diagnostic imaging technique for the identification of benign schwannoma from malignant schwannoma.But it could play an important role in the staging,restaging,and post-therapy follow-up of schwannoma.

Objective To assess the clinical value of dobutamine 201Tl stress-redistribution/nitroglycerin augmented ~(99m)Tc-metho~(99m)Tc-xyisobutylisonitrile (MIBI) gated myocardial perfusion imaging (MPI) in patients with coronary artery disease (CAD). Methods A total of 132 patients with suspected CAD underwent dobutamine 201Tl stress-redistribution/nitroglycerin augmented MIBI gated MPI and coronary arteriography (CAG) within two weeks after MPI. Percutaneous coronary interventions (PCI) were performed in 23 patients with myocardial infraction. The regional wall movement was assessed with echocardiography before and within three months after PCI. Results Taking stenosis ≥50% as standard of CAD, the sensitivity, specificity and accuracy of dual-isotopic SPECT imaging in diagnosing CAD was 93.41%, 87.80% and 91.67%, respectively. The sensitivity, specificity and accuracy in diagnosing left anterior descending artery (LAD) stenosis was 93.85%, 91.04% and 92.42%, for left circumflex artery (LCX) was 86.79%, 89.87% and 88.64%, while for right coronary artery (RCA) was 81.25%, 82.14% and 81.82%, respectively. Of all 207 myocardium segments in 23 patients with myocardial infarction, radioactivity defect was found in 113 segments on the redistributed ~(201)Tl images, and radiofilling was found in 52 (52/113) segments on the nitroglycerin augmented ~(99m)Tc-MIBI images, whereas 61 segments had no radiofilling. Taking regional wall movement improvement as the criterion of viable myocardium, the sensitivity, specificity and accuracy of identifying viable myocardium with nitroglycerin augmented ~(99m)Tc-MIBI imaging was 87.23%, 83.33% and 84.96%. Conclusion Dobutamine ~(201)Tl stress-redistribution/nitroglycerin augmented ~(99m)Tc-MIBI gated MPI is able to effectively detect CAD and viable myocardium and helpful in diagnosis and treatment of CAD.

Objective To evaluate the diagnostic value of dobutamine?stressed cardiac morphing ( CM) MPI ( CMMPI) for coronary artery disease ( CAD) in middle?aged patients. Methods A total of 97 middle?aged patients (52 males, 45 females, age 40-65 (52.98±7.74) years) from March 2011 to Febru?ary 2015 with chest distress, cardiopalmus and chest pain symptoms ( suspected CAD) were retrospectively studied. Informed consents were signed by patients before examination. All patients underwent one?day dobu?tamine?stressed/rest CMMPI and CAG within one month. The diagnostic value of CMMPI was assessed and compared with CAG. Results CMMPI revealed 42 abnormal cases and 9 normal cases in 51 cases with cor?onary artery stenosis by CAG. CMMPI found 11 abnormal cases and 35 normal cases in 46 cases with normal coronary artery by CAG. In comparison with CAG, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of CMMPI were 82.35%(42/51), 76.09%(35/46), 79.38%(77/97), 79.25%(42/53)and 79.55%(35/44) respectively. The side effect was slight and transient with an inci?dence of 82.47%(80/97). Conclusion The dobutamine?stressed CMMPI has an important value for the diagnosis of CAD and the assessment of myocardial damage in middle?aged patients.

OBJECTIVETo investigate the optimal conditions of tri-expression of CYP3A4, POR and cyt b5 in Sf 9 cells.

METHODSThe Sf 9 cells expressing CYP3A4, POR and cyt b5 were cultured in shaker flasks. The optimized conditions, including the temperature and rotation speed, the culture volume, the amount of surfactant and the culture time were studied. The expressed products in microsomes were used to metabolize the testosterone and their metabolic activity was determined.

RESULTSWhen the temperature and rotation speed of the shaker were 27 degree and 90 r/min, the cell density and culture volume were 5X105 cells/ml and 80-120 ml per 250 ml shaker flasks, respectively. When Pluronic F-68 was 0.1% and the culture time was 72 h, the condition was most suitable for culture of Sf 9 cells and expression of targeted proteins. When the ratio of the volume of three added viruses was 1:1:1, the expression condition was optimal, under which the Km, Vmax, and CLint for testosterone metabolism were 119.6 μmol/L,0.52 μmol/(min*g protein) and 4.34 ml/(min*g protein), respectively.

CONCLUSIONThe conditions of tri-expressing of CYP3A4, POR and cyt b5 have been optimized in the study and the product CYP3A4 is obtained with higher metabolic activity.

Animals ; Cytochrome P-450 CYP3A ; biosynthesis ; Cytochromes b5 ; biosynthesis ; Humans ; Insecta ; NADPH-Ferrihemoprotein Reductase ; biosynthesis ; Sf9 Cells

Objective To investigate the effects of hypoxia-inducible factor-1α (HIF-1α)expression on pathogenesis of retinopathy of prematurity (ROP) and to find new target for gene therapy.Methods After liposome-mediated small interference RNA (siRNA) transfection into rat retinal endothelial cells,the cells were cultured in medium with CoCl2-induced hypoxic condition.Expression of HIF-1α mRNA was determined by fluorenscence quantitative reverse transcription-polymerase chain reaction(RT-PCR),HIF-1α protein expression was detected by Western Blot after cocultured for 8 hours.Cell proliferation was measured with 3-(4,5)-dimethylthiazol (-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay after cocultured for 24 hours.Difference between groups was compared with independent samples t test.Results Rat retinal vascular endothelial cells were successfully transfected with siRNA.Fluorescence quantitative RT-PCR results showed that at 48 hours of transfection,the expression of HIF-1α mRNA in the interference group of siRNA1,siRNA2 and siRNA4 were 0.1620 ± 0.0147,0.2034 ± 0.0251 and 0.3049 ± 0.0165,which were 16.20％,20.34 ％ and 30.49％ of blank control group (1.0000±0.0344),and were lower than that of negative control group (0.8334±0.0242) (t=16.786,8.953 and 4.087,P＜0.05 respectively).Western Blot results showed that HIF-1α protein expression was significantly inhibited by siRNA1(0.4956 ± 0.0421 ) and siRNA2 (0.6544 ± 1.0032) comparing with blank control group (3.5105 ±0.4084) and negative control group (3.4019 ± 1.0677) (t =6.861,2.893,4.567 and 5.072,P＜0.05 respectively).As for cellular proliferation activity,(49.5±2.9) ％ and (67.4±1.2) ％ of cells growth inhibition were observed after transfection with siRNA1 and siRNA2,which were higher than those of negative control group [(15.7±1.5) ％ ] (t=2.786 and 6.904,P＜0.05).Conclusions The synthetic HIF-1α siRNA could effectively inhibit the expression of HIF-1α gene and reduce cell proliferation in rat retinal endothelial cells under hypoxic condition.RNA interference technology targeting HIF-1α might become a new strategy for gene therapy of ROP.

Objective To investigate the biodistribution and gamma imaging of 99Tcm-arginine-glutamate-threonine (RET) in nude mice bearing lung cancer xenografts and to explore its feasibility for human lung cancer imaging.Methods RET was labeled directly with 99Tcm.The binding efficiency of 99Tcm-RET with human NSCLC H1299 cells was measured.99Tcm-RET was injected via the tail vein in nude mice bearing H1299 xenografts.The mice(n=32) were sacrificed at different time points:15 min,30 min,1 h,2 h,4 h,8 h,24 h,and 48 h.Organs of interest were excised,weighted and counted by a gamma counter.The organ uptake was calculated as ％ID/g.The gamma imaging was performed on 3 mice at 0.5,1,2,4,4.5,5,6 h after intravenous injection of 4.81 MBq 99Tcm-RET.Results The radiolabeling yield of 99Tcm-RET was (93.15±2.02)％,and the binding efficiency of 99Tcm-RET with H1299 cells was (3.56±0.37)％.At 4 h after injection,the uptake of tumor,liver and spleen was (4.96±1.05) ％ID/g,(15.89±1.84) ％ID/g and (10.83±1.66) ％ID/g and the T/NT was 5.70±0.21 for the heart and 12.40±0.11 for the blood.The tumor in nude mice could be best visualized at 4.5-6.0 h.Conclusion 99Tcm-RET might have potential to serve as a lung cancer imaging agent.

Though all the marketed drugs of dipeptidyl peptidase IV inhibitors are structurally different, their inherent correlation is worthy of further investigation. Herein we rapidly discovered a novel DPP-IV inhibitor 8g (IC50 = 4.9 nmol.L-1) which exhibits as good activity and selectivity as the market drugs through scaffold hopping and drug splicing strategies based on alogliptin and linagliptin. This study demonstrated that the employment of classic medicinal chemistry strategy to the marketed drugs with specific target is an efficient approach to discover novel bioactive molecules.

The enantioseparations of 38 racemates on Chiralcel OD, Chiralpak AD, Chiralpak IA and(S, S)-Whelk-01 were presented by HPLC. Those enantiomers come from the amines, alcohols, ethers, ketones, aromatic derivatives, heterocyclic compounds, amide acids and medicines etc. With the mobile phase of n-hexane)/isopropanol(90∶ 10, V/V), n-hexane/isopropanol/trifluoracetic acid(90∶ 10∶ 0.2, V/V) or n-hexane/isopropanol/triethylamine(90∶ 10∶ 0.2, V/V), over 70% enantioseparations were obtained for OD, AD and IA columns). The order of enantioseparation selectivity for four columns was OD>AD>IA>(S,S)-Whelk-01, and among those columns there was a big chiral discriminating complementarity. This investigation was useful for choosing chiral columns to separate chiral compounds.

Objective To study the expressions of programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1) in liver injury of acute liver failure (ALF) in rats and the role of PD-1/ PD-L1 in liver inflammatory injury. Methods SD rats were divided into two groups: 6 in normal group and 30 in ALF model group. The ALF models in rats were induced by D-galactosamine (D-Gal). The sera and hepatic tissue samples were collected at 12, 24, 48, 72 and 120 hours after D-Gal injection. Expressions of PD-1 mRNA and PD-L1 mRNA in hepatic tissue samples were detected by reverse transcription-polymerase chain reaction (RT-PCR). Comparison of measurement data between groups was done by t test. Correlation test was performed using Pearson linear correlation analysis. Results The levels of alanine animotransferase (ALT) and aspartate animotransferase (AST) at 12 h of D-Gal injection were (217. 3±33. 7) U/L and (397. 2± 101.3) U/L, respectively,which were both significantly higher than those in normal group [(30. 5 ±3. 1) U/L and (78. 6±4.2) U/L, respectively; t=-8. 921 and -6. 121, respectively; both P＜0.01] and peaked at 48 h.The expression of PD-1 mRNA in model group at 12 h (0. 385±0. 074) was significantly higher than that in normal group (0. 097±0.009) (t= -7. 725, P＜0.01) , and peaked at 48 h (0. 927±0. 132),then decreased obviously at 72 h. The expression of PD-L1 mRNA in the liver tissue of normal rats was very little, while that in model group was increased gradually over time, then peaked at 48 h (0. 593±0. 105; t =- 10. 076, P＜0. 01). The expressions of PD-1 and PD-L1 were positively correlated with ALT level (r=0. 807 and 0. 792, respectively; both P＜0. 01). Conclusion The expressions of PD-1/PD-L1 may play an important role in liver inflammatory injury in rat model of acute liver failure.