OBJECTIVE:To establish a method for simultaneous determination of the contents of geniposide and liquiritin in Xinshenning tablets. METHODS:HPLC was performed on the column of Inertsil ODS-3 with mobile phase of acetonitrile-water (gradient elution) at flow rate of 1.0 ml/min,the column temperature was 25 ℃,detection wavelength was 238 nm and the vol-ume was 10 μl. RESULTS:The mass concentration range was 16.3-407.9 μg/ml for geniposide(r=0.999 9) and 7.4-185.9 μg/ml for liquiritin(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.63%;average recoveries were 99.05%(RSD=1.28%,n=9)and 98.00%(RSD=1.84%,n=9),respectively. CONCLUSIONS:The established method is accu-rate,sensitive and simple,and can be used for the contents determination of geniposide and liquiritin in Xinshenning tablets.

Objective:To investigate the effect of sorafenib on the proliferation of human oral cancer TCA8113 cells and to explore the underlying mechanisms.Methods:Mter treated with sorafenib at 2.5,5,10,20 μg/ml respectively for48 h,TCA8113 cell proliferation was examined by MTT and colony formation assay.Western blotting was employed to examine the p38MAPK expression in the cells.TCA8113 cells were pretreated with 10 μmol/L of SB203580 (a specific inhibitor of p38MAPK) for 30 min,and then by different concentrations of sorafenib for 48 h,cell proliferation was tested by MTT assay.Results:Sorafenib significantly inhibited the proliferation of TCA8113 cells in a concentration dependent fashion.Sorafenib also remarkably promoted the activation of p38MAPK of the cells.SB203580 significantly alleviated soiafenib induced TCA8113 cell viability decrease.Conclusion:Sorafenib can inhibit the proliferation of TCA8113 cells,which may be related to the activation of p38MAPK.

Objective To determine the expression of Caspase 8 and phospho-Akt(p-Akt)in condyloma acuminatum(CA)lesions, and to evaluate their significance. Methods Skin lesion samples were collected from 30 patients with CA, cancer tissue samples from 20 with cervical cancer, and normal skin samples from 20 healthy controls. All the samples were subjected to paraffin embedding. An immunohistochemical study was conducted to determine the expression and distribution of Caspase 8 and p-Akt in the above samples. Results The expression rate of Caspase 8 was significantly lower in CA lesions (23.33%)than in normal skin samples(90%, P < 0.01)and cervical cancer lesions(80%, P < 0.001). Moreover, the expression rate of p-Akt in CA lesions(93.33%)was significantly higher than that in the normal skin samples(90%, P<0.001), but lower than that in the cervical cancer lesions(95%, P<0.001). No significant correlations were observed between the expression of Caspase 8 and p-Akt in either CA lesions or normal skin samples. However, the expression of Caspase 8 was positively correlated with the expression of p-Akt in cervical cancer lesions(r=0.369, P<0.05). Conclusion Both suppressed apoptosis initiation of Caspase 8 and anti-apoptotic effect of p-Akt may be involved in the occurrence and development of CA.

Objective To investigate the regulatory effects of miR-145 on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively.After additional culture for different durations,real-time PCR was performed to determine the expression level of miR-145,MTS assay to estimate cell proliferation,and flow cytometry to detect cell apoptosis and cycle.Luciferase assay,real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145.Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00 ± 1.21 folds compared with NC mimic-transfected cells (t =115.90,P ＜ 0.0001).The transfection with miR-145 mimics significantly inhibited the proliferation of HaCaT cells (F =8.76,P =0.008),and the inhibitory effect significantly varied with the duration (24-96 hours) of culture after transfection,with no interaction effect between the transfection with miR-145 mimics and culture duation (F =1.21,P =0.18).Compared with NC mimic-transfected cells,those transfected with miR-145 mimics showed a significant increase in the proportion of early apoptotic cells (18.9％ ± 4.1％ vs.4.3％ ± 1.2％,t =7.126,P ＜ 0.01),late apoptotic cells (9.3％ ± 2.3％ vs.3.6％ ± 1.6％,t =12.38,P ＜ 0.01),G1-phase cells (85.83％ ± 5.2％ vs.62.08％ ± 6.23％,t =11.78,P =0.007),but a significant decrease in the percentage of G2-phase cells (6.26％ ± 1.2％ vs.19.36％ ± 3.45％,t =7.610,P =0.017) and S-phase cells (7.91％ ± 1.3％ vs.18.56％ ± 5.23％,t =7.230,P=0.019).As luciferase assay showed,luciferase activity was significantly lower in HaCaT cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-wild carrying the wild-type 3'UTR of NRAS than in those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-wild (t =11.09,P =0.008),but similar between cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-mut carrying the mutant-type 3'UTR of NRAS and those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-mut (P ＞ 0.05).Real-time PCR and Western blot revealed that the overexpression of miR-145 mimics had no significant effect on NRAS mRNA expression (P ＞ 0.05),but significantly inhibited NRAS protein expression (1.52 ± 0.07 vs.0.20 ± 0.02,t =28.43,P＜ 0.01).Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS.

AIM:To investigate the effect of genistein on the proliferation of human oral cancer TCA 8113 cells and to explore the underlying mechanisms .METHODS:The cell proliferation was examined by MTT assay , cell counting and colony formation assay .Western blotting was employed to examine the protein levels of vascular endothelial growth fac -tor (VEGF), extracellular signal-regulated kinase (ERK) and p-ERK.RESULTS: Genistein significantly inhibited the proliferation of TCA8113 cells in a concentration-dependent fashion .Moreover , genistein dose-dependently decreased the protein levels of VEGF, ERK and p-ERK.The expression of VEGF was also blunted by U 0126, a specific inhibitor of ERK.U0126 and axitinib, a VEGF receptor antagonist , both significantly inhibited the proliferation of TCA 8113 cells. CONCLUSION:Genistein inhibits the proliferation of TCA8113 cells, which may be related to its inhibitory effect on ERK expression and activation , thus subsequently decreasing the expression of VEGF .

To explore evaluation of echocardiography (ECG) for early change of right ventricular function in aged patients with chronic obstructive pulmonary disease (COPD).Methods :A total of 45 aged patients with mild‐moderate stable COPD were selected as COPD group , and another 45 healthy volunteers were regarded as healthy control group .Two‐dimensional ultrasound and Doppler ultrasound indexes were measured and compared be‐tween two groups ,and their correlation were analyzed .Results :Compared with healthy control group ,there were significant rise in right ventricular transverse diameter [RVTD ,(28.03 ± 2.83) mm vs.(37.55 ± 5.44) mm] ,right ventricular wall thickness [RVWT ,(4.79 ± 0.44) mm vs.(5.04 ± 0.66) mm] ,right ventricular myocardial per‐formance index [RVMPI , (0.38 ± 0.06 ) vs .(0.54 ± 0.12 )] , tricuspid regurgitation pressure gradient [TRPG , (9. 50 ± 1. 93) vs.(36.87 ± 6.27 )] , tricuspid diastolic peak E／tissue Doppler tricuspid annular diastolic e′[E／e′, (4. 24 ± 0. 72) vs.(6.48 ± 1. 51)] and significant reductions in right ventricular fraction area change [FAC ,(53. 35 ± 5.41)% vs.(40.21 ± 7.24)%] ,tricuspid annular plane systolic excursion [TAPSE ,(20. 78 ± 2.64) vs.(14. 15 ± 2.44)] ,tricuspid early／late diastolic peak flow velocity [E／A , (1.17 ± 0.19 ) vs.(0.73 ± 0.14 )] , and tissue Doppler tricuspid annular systolic peak velocity [DTIs′,(13. 71 ± 1.25) vs.(12.92 ± 1. 70)] in COPD group , P＜0.05 or ＜0.01. Pearson correlation analysis indicated that TRPG was significant positively correlated with RVTD , RVMPI and RVWT ( r＝0. 440～0.830 , P＜0. 01 all) ,and significant inversely correlated with FAC ,TAPSE and DTIs＇( r＝ －0.615～ －0.561 , P＝0.001 all).Conclusion :RVTD ,FAC ,TAPSE ,tricuspid E／A ,E／e＇,RVMPI and TRPG etc .can be used to assess early change of right ventricular function .

Objective To explore the specific plasma exosomal protein expression in Coronary Heart Disease(CHD)with phlegm-turbidity syndrome(PTS),so as to provide biological reference for the diagnosis of CHD with PTS.Methods A total of 20 CHD patients with PTS and 20 healthy subjects at the same period were included.Plasma exosomes were extracted from all subjects using the qEV size exclusion method.First,10 CHD cases with PTS and 10 healthy subjects were randomly selected,and differentially expressed proteins between the two groups were screened using label free non-targeted protein quantification analysis.Then,MRM targeting protein labeling technique was applied to verify the differentially expressed proteins in the remaining subjects.Results Compared with healthy controls,214 differentially expressed plasma exosomal proteins(61 up-regulated and 153 down-regulated)were found in CHD patients with PTS,mainly related to cholesterol metabolism,complement and coagulation cascade,and immune effects.ANXA6,C4BPB,F8,CFB,APOE,C9,and CLU proteins were further validated by MRM targeting protein.Conclusion CHD patients with PTS had differences in plasma exosomal protein expression from healthy controls,and the differential proteins are mainly related to cholesterol metabolism,complement,and the coagulation cascade.