Objective To determine the content of cholesterol in Gekko Gecko.Methods High performance liquid chromatography was performed on Hypersil ODS2 (4.6 mm?150 mm,5 ?m).The chromatorgraphic conditions were as follows:methanol as mobile phase,flow rate being 1.0 mL/min and detecting wavelength at 208 nm.The column temperature was 40 ℃.Results A good linearity of cholesterol was shown in range of 0.885～14.000 ?g (r=0.999 5),the average recovery was 99.65%,RSD=3.45% (n=6).Conclusions This method is simple,rapid and accurate,and may provide a reliable way for the quantification of cholesterol in Gekko Gecko.

The baculovirus expression system has been used extensively for the expression of recombinant proteins in insect cells.Recently,recombinant baculoviruses containing mammalian cell-active promoter element have been used to transduce a broad spectrum of primary and established mammalian cells,including non-hepatic cells.Recombinant baculoviruses have been used successfully for transient or stable gene delivery in mammalian cells in vitro,while the efficiency of delivering gene in vivo is inhibited obviously by complements,but efforts have been made to overcome the problems,for instance,VSV-G-pseudotyped baculoviruses display complement resistance.The mechanism of the transduction is still not clearly understood,though many researches have been done.The baculovirus is able to replicate only in insect cells,but it is incapable of initiating replication cycle in mammalian cells,which guarantees high biosafety of this gene delivery system.In addition,this system is easily manipulated and able to carry large inserts.These attributes will undoubtedly lead to the increased application and continued development of this system for highly effective gene delivery into mammalian cells.

Background The measurement of corneal Q value is essential for corneal refractive surgery and calculation of intraocular lens during cataract surgery.Topolyzer was often used for the measurement of Q value,and recently Topcon KR-1W and iTrace were applied in ophthalmology.However,whether the measured values are interchangeable is unclear.Objective This study was to assess the difference and consistency of corneal Q values measured by Topcon KR-1W,iTrace and Topolyzer.Methods Corneal Q values were measured on 100 right eyes of 100 healthy subjects under the approval of Ethic Committee of the Sixth Hospital Affiliated to Shanghai Jiaotong University and informed consent of each subject from November to December in 2014 with Topcon KR-1W,iTrace and Topolyzer.Three valid measurements were obtained for each device,and the average values from each device were calculated.The difference of the outcomes among the instruments was compared by repeated measures analysis of variance (ANOVA),and the consistency among the outcomes from different apparatus was analyzed by Bland-Altman plots.Results The mean corneal Q values were-0.184-±0.112,-0.117±0.167 and-0.269±0.117 from Topcon KR-1W,iTrace and Topolyzer,respectively,with a significant difference among them (P ＜ 0.001).The measured Q value by Topcon KR-1W was 0.085±0.010 larger than that by Topolyzer,and the Q values by iTrace was 0.152± 0.014 larger than that by Topolyzer,while the Q values obtained by Topcon KR-1W was 0.067±0.016 smaller than that by iTrace (all at P＜0.05).The 95％ confidence interval of the values between Topcon KR-1W and iTrace,Topcon KR-1W and Topolyzer,iTrace and Topolyzer were-0.106 to-0.028,0.060 to 0.109 and 0.118 to 0.186,respectively.Bland-Altman plots showed that 6％,6％ and 5％ values were outside of 95％ agreement of limit (LoA) between Topcon KR-1W and iTrace,iTrace and Topolyzer or KR-1W and Topolyzer,respectively,with the maximal differences of 0.28,0.43 and 0.38.Conclusions Corneal Q values measured by Topcon KR-1W and iTrace are larger than those measured by Topolyzer.Due to the poor agreements among the corneal Q values by the 3 kinds of devices,they are not interchangeable in clinical applications for the measurement of corneal Q value.

Aim To investigate the effects of dalbinol on proliferation and apoptosis of human colon cancer HCT1 16 cells and its mechanisms.Methods Anti-proliferative effect of dalbinol was evaluated by MTT assay.The morphological changes of apoptosis were observed by Hoechst33342 staining.Apoptotic rate and ROS generation were analyzed by flow cytometry.The related proteins of Wnt/β-catenin pathway and the ap-optosis-associated proteins expression were measured by Western blot.Results The growth of HCT1 16 treated with dalbinol was inhibited in a dose and time dependent manner with IC50 (4.8 ±0.53 ),(2.5 ± 0.43)and (0.6 ±0.22)μmol·L-1 at 24,48 and 72 h,respectively.Typical morphological changes of ap-optosis such as cell shrinkage,karyopyknosis and nu-clear condensation were observed by Hoechst33342 staining.Meanwhile,the apoptotic rate and intracellu-lar ROS generation of dalbinol were both increased dose-dependently. Western blot results showed that dalbinol could activate the expression of cleaved Caspase-3 and cleaved PARP by decreasing anti-apop-totic protein levels such as Bcl-2 and Mcl-1 and in-creasing pro-apoptotic protein levels such as Bax and Bim,which induced further apoptosis.Moreover,dal-binol can reduce the protein expression of the total and nuclear β-catenin,but not cytoplasmic β-catenin by suppressing the protein expression of Dvl-2 and GSK-3β(pS9 ),as well as its target proteins c-Myc and Sur-vivin.Conclusion dalbinol can induce apoptosis in colon cancer HCT1 16 cells by upregulating the intra-cellular ROS generation and suppressing Dvl/GSK-3β/β-catenin pathway.

ObjectiveTo study the efficacy of transcatheter arterial chemoembolization (TACE) after liver resection for hepatocellular carcinoma (HCC) with tumor thrombus in the main trunk and/or first branch of portal vein,and to clarify prognostic factors affecting survival.Methods From 2005 to 2009,there were 358 consecutive patients with HCC who underwent surgical resection in our Department.In 55 patients (15 ％),portal vein tumor thrombus (PVTT) was found intraoperatively or postoperatively during histopathological examinations to involve the first portal branch,main portal trunk,or contralateral portal branch.In this retrospective study,these 55 patients were divided into two groups:Group A,29 patients received postoperative TACE,and Group B,26 patients who did not receive TACE.The clinical data and survivals were compared between the two groups.Prognostic factors were indentified using univariate analysis,followed by multivariate regression analysis using the Cox proportional hazards model.ResultsThere were no significant differences in the demographic clinical data between Group A and Group B.The overall 1-,2- and 3-year survivals for the 55 patients were 63.3 ％,51.4 ％ and 43.5 ％,respectively.The accumulative 1-,2- and 3-year survivals for group A were 71.4 ％,60.1 ％ and 50.1 ％,respectively.The corresponding figures for group B were 56.7％,21.7％ and 10.4％,respectively.Multiple tumors,intrahepatic metastases,hepatic vein thrombus,and invasive type of tumor thrombus were found to be risk factors for short-term survival on univariate analysis,while the latter 3 factors were further found to be significant prognostic factors in the Cox proportional hazards model.Postoperative TACE was shown to be a significant factor in both univariate and multivariate analyses.ConclusionLiver resection was beneficial for some patients with portal vein tumor thrombus.Postoperative TACE further improved the prognosis and prolonged survivals in these patients.

Antigens, CD20 ; metabolism ; Epstein-Barr Virus Infections ; virology ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Lymphoma, Large B-Cell, Diffuse ; pathology ; virology ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; virology

Background The neovascular form of the disease usually causes severe vision loss in a number of eye diseases.Special-domain optical coherence tomography (SD-OCT) provides high-quality in retinal imaging and the possibility of the measurement in vivo.Objective This study aimed to investigate the feasibility of SD-OCT dynamically measuring choroidal neovascularization (CNV).Methods CNV was induced in 30 left eyes of 30 clean Brown Norway(BN)rats by retinal photocoagulation with the laser parameter as follows: wavelength 532 nm,exciting power 200 mW,spot diameter 100 μm and irradiating time 50 ms.Bubble or less retinal bleeding was thought as Brunch membrane breakage and CNV model establishment.Fundus fluorescein angiography(FFA) was performed to determine the establishment of CNV model and scored based on the fluorescein leakage on 3,7,14,21 days after photocoagulation.Meanwhile,CNV memberane thickness (CMT) was dynamically measured in vivo as the maxiume value from retinal inner limiting membrane through choroidal vessel layer in various time points.Histopathologic examination was used in the 14th day to evaluate and verify the result of SD-OCT.The right eyes were as controls.Results FFA examination showed that disc-like leakage of fluorescein appeared in 7 days and extended in 14 days after photocoagulation with the scores of 1.6±0.4,2.5±0.6 and 2.4±0.5 in 7,14 and 21 days,showing a significant difference among them(F=13.11,P＜0.01).The fluorescein leakage score was significantly higher in 14 and 21 days than that of 7 days(both P＜0.05).CMT measured by SD-OCT was(76.33±10.09),(102.03±14.21)and(98.03±13.76) μm in 7,14 and 21 days after photocoagulation respectively,with a significant difference among 3 time points (F=23.25,P＜0.01),and that in 14 and 21 days was significantly declined in comparison with 7 days(both P＜0.05).The results of SD-OCT showed a consistent tendency with that of FFA.Histopathological examination showed CNV formation in 14 days after photocoagulation.Conclusions Experimental CNV model was successfully induced by laser photography.SD-OCT technology allows excellent visualization of CNV in vivo.

ObjectiveTo evaluate the efficacy and safety profiles of enteeavir (ETV) in patients with advanced schistosomiasis and hepatitis B virus (HBV) co-infection.Methods Totally sixty patients with advanced schistosomiasis and HBV co-infection were enrolled in this study.The patients were divided into ETV treatment group (n=30) and rhubarb treatment group who refused to receive antiviral treatment (n=30).The patients were treated with ETV or rhubarb thelepus ball on the basis of routine supportive therapy for 52 weeks.The hepatic fibrosis markers (e.g.hyaluronic acid,type Ⅲ procollagen,type Ⅳ collagen,laminin and fibronectin),alanine transaminase (ALT),HBV DNA,Child-Pugh score between two groups were compared.Intention to treat (ITT) population was used for analysis.The measurement data and the enumeration data were analyzed by t test and x2 test,respectively.ResultsAfter 52-week treatment,the hepatic fibrosis markers (hyaluronic acid,type Ⅲ procollagen,type Ⅳ collagen,laminin and fibronectin) were significantly improved in ETV treatment group compared to the rhubarb treatment group (t =3.952,3.765,3.857,3.122 and 3.735,respectively; all P＜0.05),and the fibrosis of liver tissue in ETV treatment group was significantly improved compared with rhubarb treatment group (x2 =11.207,P＜0.05).The ALT level,HBV DNA,Child-Pugh score after 52-weeks treatment in ETV treatment group were statistically reduced compared with rhubarb treatment group (t =3.287,4.382 and 3.872,respectively; all P＜0.05),meanwhile,the ALT normalization rate and HBV DNA undetectable rate were significantly increased in ETV treatment group (x2 =17.376 and 39.095,respectively; both P＜0.05).In addition,no obvious adverse reaction was observed during ETV treatment.Conclusion Entecavir is safe and effective in patients with advanced schistosomiasis and HBV co-infection.

Objective To investigate the effects of simvastatin (SV) on the proliferation,differentiation and apoptosis of human promyelocytic leukemia cell line NB4.Methods NB4 cells were incubated with SV at different concentration with or without all-trans retinoic acid (ATRA),and NB4 cells without any treatment were taken as normal control.Cells of different groups were collected at 24 h,48 h and 72 h after incubation for further detection.Morphological changes by Wright stain were performed.MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the surface CD11b expression levels,the early stage apoptosis ratio and cell necrosis ratio.Results Treated with 15 μ mol/L SV,10 μ mol/L SV and 5 μ mol/L SV respectively,with the NB4 cells growth,the cell inhibition rates gradually increased (F =7.15,P =0.000),as well as CD11b expression levels (F =3.41,P =0.014) and AnnexinVexpression levels (F =43.38,P =0.000).Furthermore the NB4 cells treated with 15 μ mol/L SV exhibited the most significant changes with cell inhibition rate of 0.96±0.02,CD11b expression level increased to (62.41±6.37) ％ and AnnexinV expression level increased to (87.38±2.94) ％ after 72 h incubation.Combination of 15 μmol/L SV with 0.5 μmol/L ATRA displayed obvious interaction for increasing CD11b expression levels (F =4.093,P =0.025),while no significant interaction for cell inhibition rates and Annexin V expression levels were observed.After 72 h incubation,the CD11b expression levels (89.46±9.13) ％ in NB4 cells treated with 15 μ mol/L SV in combination with 0.5 μ mol/L ATRA were significantly higher than those treated with ATRA (71.27±7.27) ％ and SV (62.41±6.37) ％ (t =2.71,P =0.054; t =4.37,P =0.017)' solely.Conclusion Simvastatin in vitro inhibits NB4 cell proliferation,promotes cell apoptosis,and synergistically induces cell differentiation with ATRA dose-dependently in vitro,which indicates that SV may have the effect of synergistic anti-promyelocytic potency with ATRA.

Objective To investigate the effect of simvastatin (SV) in combination with cytosine arabinoside (ARA-C) on the proliferation and apoptosis of K562 cells. Methods Human K562 cells were incubated with SV and cytosine arabinoside alone or in combination and K562 cells without any treatment were taken as normal control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detections. Morphological changes by Wright stain were performed. MTT method was used to assay the growth inhibition rate and cytoflowmetry was used to detect the early stage apoptosis ratio and cell necrosis ratio. Results Compared with Ara-C group and SV group, cells in the group treated with SV combined with Ara-C showed obvious karyopyknosis,apoptosis bodies formation and significant cell growth inhibition, which were positively correlated with culture time. Combination of 15 μmol/L SV and Ara-C showed the most significant cell growth inhibition with a inhibition rate of (72±1) % at 72 h of culture, as was significantly higher than that of 15 μmol/L SV group (45±2) % and 20 μmol/L Ara-C group (44±0) % (P ＜0.01),furthermore, combination of 15 μmol/L simvastatin and Ara-C showed synergistic inhibition with Q value of 1.24 and 1.19 at 24 h and 48 h in each. The apoptosis rates at early stage (AnnexinV) detected by flow cytometry in 20 μmol/L, 15 μmol/L and 10 μmol/L SV treated K562 cells were significantly higher than that in normal K562 cells (P ＜0.01), as were positively correlated with culture time and SV dose (P ＜0.05). There were no significant difference of early apoptosis rate between the 20 μmol/L SV and 15 μmol/L SV groups (P ＞0.05), yet the very two were both higher than that of 10 μmol/L SV group (P ＜0.05). There were no statistic differences of late apoptosis rate (PI) amongdifferent treated groups (P ＞0.05). Conclusion SV inhibited K562 cell proliferation and induced cell apoptosis in vitro, and combination of SV and Ara-C exhibited obvious synergistic inhibition and apoptosis, which may increase the sensitivity of K562 cell to chemotherapy. SV at 15 μmol/L may be the best concentration for K562 cells in vitro.