Objective To explore the function of fluorescent probe JC-1 in detecting the changes of mitochondrial membrane potential(△?m)in early apoptotic cells.Methods After 2-ME was used to induce MUTZ-1 cell apoptosis,cells were dyed with fluorescent probe JC-1,and then the changes of △?m in the early stage of apoptotic cells were analyzed by flow cytometry or detected under fluorescent microscope. Results The control cells with high △?m are those forming JC-1 aggregates in the inner membrane of mitochondria,thus showing orange-red fluorescence.2-ME caused decrease of △?m in MUTZ-1 cells,in which JC-1 maintains monomeric form,thus showing only green fluorescence.The decreases of △?m were in a time-dependent manner,which were significantly higher than those in control group(P

AIM To optimize the combined enzymatic extraction for alkaloids and polysaccharides from Dendrobium nobile Lindl..METHODS With enzyme consumption,enzymolysis temperature,enzymolysis time and solidliquid ratio as influencing factors,contents of dendrobine,total alkaloids and polysaccharides as evaluation indices,orthogonal test was applied to optimizing the combined enzymatic extraction.RESULTS The optimal conditions for papain extraction were determined to be 0.10 g for enzyme consumption,45 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶ 50 for solid-liquid ratio,the contents of dendrobine,total alkaloids and polysaccharides were 3.495 5,4.341 8 and 35.898 7 mg/g,respectively.The optimal conditions for cellulase extraction were determined to be 0.30 g for enzyme consumption,50 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶40 for solid-liquid ratio,the contents of three constituents were 3.514 8,4.351 3 and 36.331 2 mg/g,respectively.The optimal conditions for pectinase extraction were determined to be 0.45 g for enzyme consumption,55 ℃ for enzymolysis temperature,2.5 h for enzymolysis time,and 1 ∶ 40 for solid-liquid ratio,the contents of three constituents were 3.524 4,4.452 8 and 26.324 2 mg/g,respectively.CONCLUSION This stable and reliable method can be used for the rapid combined enzymatic extraction for alkaloids and polysaccharides from D.nobile.

Background Researches showed that as the non-optical factors,cognitive has certain influence on the regulating system.So accurately experimental design is one of the key steps that evaluates the non-optical factors on regulating system.Objective The present study was to investigate the influence of presenting pattern of target and watching way on the level and amplitude of accommodative fluctuate and to analyze the effect of focus gaze of cognitive on regulating system and the relationship between focus gaze condition under near work and the development of myopia.Methods This study complied with Declaration of Helsinki,and the permission of Ethic Committee and written informed consent was obtained before entering in this trial.Thirty healthy volunteers were included with the mean age (24.80 ± 1.98) years old,equivalent refractive diopter (-1.92 ± 2.02) D and mean cylinder (-0.19±0.58) D.The presenting pattern of the targets was designed as focus gaze and relaxed gaze.The accommodative response and accommodative fluctuation in the complete corrected right eyes for the different targets at the 40 cm under the gazing state was recorded with Grand Seiko WAM 5500 automatic infrared refractor in the experiment.Results The mean accommodative response value was (1.86±0.26) D under the focus gaze and (1.27±0.39) D under the relax gaze,showing a statistically significant difference (t=-8.052,P=0.000).The mean fluctuate value was(0.17±0.06) D under the focus gaze,with a significant lowing in comparison with (0.28±0.17) D under the relax gaze (t =3.600,P =0.001).Conclusions These results demonstrate that the different presenting patterns of sighting target and watching ways of the subjects affect accommodation system.The accommodative response was relatively more accurate with a smaller microwavc moving under the focus gaze condition.

Survivin a novel member of the inhibitor of apoptosis protein family, is overexpressed in most types of cancer but not in normal differentiated adult tissues. Its mRNA expression levels among hematogical malignancies are characteristic in each type, subtype and distinctive in different phases of disease, making it a reliable diagnostic marker for clinical stages. Recently, researches indicate that high levels of survivin expression are associated with a poor prognosis and may be involved in tumor resistance to multiple chemotherapeutic drugs. In addition, experiments demonstrate that leukemic vaccination with DC pulsed with survivin antigen in vitro inhibit the proliferation of leukemic cells. Furthermore, when transferred survivin antisense oligodeoxynucleotide or dominant-negative mutant survivin into, malignant cells can be induced apoptosis mediated by downregulation in survivin expression. These findings suggest that survivin may serve as a potential target for biological strategies against hematological neoplasms. This review focuses on expression of survivin in hematological malignancies, effects of survivin on drug-resistance and prognosis of hematological malignancies, and application of survivin in the treatment of hematological malignancies.
Apoptosis ; genetics ; Biomarkers, Tumor ; genetics ; Gene Expression Regulation, Neoplastic ; Hematologic Neoplasms ; genetics ; pathology ; Humans ; Inhibitor of Apoptosis Proteins ; Leukemia ; genetics ; pathology ; Lymphoma ; genetics ; pathology ; Microtubule-Associated Proteins ; genetics ; Myelodysplastic Syndromes ; genetics ; pathology ; Neoplasm Proteins ; genetics

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; therapeutic use ; DNA-Binding Proteins ; genetics ; metabolism ; Drug Therapy, Combination ; Endonucleases ; genetics ; metabolism ; Female ; Fluorouracil ; pharmacology ; therapeutic use ; Humans ; Male ; Middle Aged ; Organoplatinum Compounds ; pharmacology ; therapeutic use ; RNA, Messenger ; drug effects ; metabolism ; Statistics as Topic ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; Survival Analysis ; Treatment Outcome

OBJECTIVETo study the effect of 2-methoxyestradiol(2-ME) on telomerase activity expression and apoptosis in human myelodysplastic syndrome cells line MUTZ-1.

METHODSMUTZ-1 cells were incubated with different concentrations of 2-ME, apoptosis rate and cell cycle were measured by flow cytometry (FCM). Telomerase activity in MUTZ-1 cells was examined by telomeric repeat amplification protocol-Enzyme linked immunosorbent assay (TRAP-ELISA).

RESULTSThe FCM analysis showed that cells in G0/G1 phase and S phase were decreased, while in G2/M phase increased after exposed to 1,2 and 4 micromol/L of 2-ME for 12 hours (P < 0.05). 1 and 2 micromol/L of 2-ME had no notable effect on MUTZ-1 cells as compared with the control group (P > 0. 05). Cells incubated with 1, 2 and 4 micromol/L of 2-ME for 36 hours were induced apoptosis, the percentage of apoptosis was between (12.87 +/- 0.86)% and (21.82 +/- 1.71)% with a dose- and time- dependent manner. Telomerase activity was significantly inhibited in these concentration and negatively correlated with cell number in G2/M phase (r = -0.979, P = 0.021) and increased apoptosis (r = -0.970, P = 0.030 ), respectively. Moreover, the inhibition effect of telomerase activity was enhanced in a dose- and time- dependent manner.

CONCLUSIONS2-ME-induced apoptosis and inhibition of telomerase activity provide a possible mechanism for explaining the 2-ME's anticancer activity.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; Myelodysplastic Syndromes ; enzymology ; pathology ; Telomerase ; metabolism

OBJECTIVETo investigate the effect of ischemic postconditioning on the expression of rat myocardium matris metalloproteinase-2 (MMP-2) induced by ischemia/reperfusion (I/R) and relationship between its expression and interstitium and the effect on left ventricular function.

METHODSTwenty-four rats were randomly divided into 3 groups (n = 8): sham control (SC) group, ischemic/reperfusion (I/R) group and ischemic postconditioning (IPTC) group. The left ventricular peak systolic pressure and its derivate (+/- dp/dt) were calculated; The amount of myocardium collagenous were determined; The vitality of superoxide dismutase (SOD) and content of malondialdehyde (MDA) of plasma were detected; The activity of myocardium MMP-2 was measured by Western blot and RT-PCR.

RESULTSAs compared with I/R group, IPTC could lower the expression of MMP-2, ameliorate left ventricular function and increase the content of myocardium collagenous. In the meantime, the vitality of superoxide dismutase (SOD) of plasma were greatly enhanced and the content of malondialdehyde (MDA) of plasma were reduced in IFC group.

CONCLUSIONProtective effect of IPIC on myocardium may be due to reduce free radical, lower expression of MMP-2 and protect myocardial interstitium. MMPs plays an important role in the myocardial protection provided by IPTC.

Animals ; Collagen ; metabolism ; Ischemic Postconditioning ; Malondialdehyde ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Myocardial Reperfusion Injury ; enzymology ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Objective To investigate the expression of Toll-like receptor(TLR) 9 of circulating plasmacytoid dendritic cell (pDC) and analyze the frequency and interferon (IFN)-?production of circulating pDC during hepatitis B virus(HBV) infection.Methods Peripheral blood was collected from 69 HBV-infected patients,including 14 cases of asymptomatie HBV infection,30 cases of chro- nic hepatitis B(CHB),25 cases of HBV-related liver cirrhosis,and 21 healthy blood donors as con- trols.Flow cytometry was used to analyze the frequency of circulating pDC and the mean fluorescence intensity(MFI) of TLR9.Fresh peripheral blood mononuclear cells(PBMC) were stimulated by CpG ODN 2216 for 24 h in vitro.IFN-?in the supernatant was measured using enzyme-linked immunosorbent assay(ELISA) to analyze the frequency and IFN-?production of pDC during HBV in- fection.Data were analyzed with SPSS 13.0 for windows.Results Compared with healthy controls (62.6?10.7),the MFI of TLR9 of patients with asymptomatic HBV infection,those of CHB and HBV-related cirrhosis were significantly reduced (P

Objective To analyze the ultrasonographic features of retroperitoneal ifbrosis (RPF). Methods Totally 13 patients with retroperitoneal ifbrosis from February 2000 to October 2012 in the Long Gang central Hospital of Shenzhen were retrospectively analyzed. Results In all patients who underwent ultrasound examination, there were ten cases of idiopathic RPF and three cases of secondary RPF with abdominal tumors. In 11 cases, the masses were hypoechoic locating at retroperitoneum and surrounding the abdominal aorta without deifnitive margin. One case showed hypoechoic mass with clear boundary. In ten cases, the internal echogenicity of masses were uniform. In two cases, the internal echogenicity of masses were uneven with a small amount of ifbrous separator with slightly higher echogenicity. No blood was found in all masses. The encasement of inferior vena cava was found in three casesand the masses extended to iliac arteries in three cases. Hydronephrosis could be found in 11 patients (84.6%) and ureter dilatation was found in ten cases. Ureteral localized stenosis were found in two cases. Conclusion Ultrasonography is a preferred imaging method in diagnosing RPF.

Carnosic acid (CA) is the main phenolic diterpenoid active ingredient in plants such as rosemary and sage, and has antiviral, antioxidant, anti-inflammatory effects and so on, however, its antiviral activity against influenza virus infections was not reported. In this study, antiviral activities against influenza A virus infections of three main bioactive ingredients from rosemary, including rosmarinic acid, CA and ursolic acid, were evaluated using virus titer titration assay, and CA showed remarkable inhibition on influenza H5N1 replication in A549 cells. The antiviral activity of CA was further confirmed and its mechanism of action was investigated using the indirect immunofluorescence assay (IFA), Western blot and real-time fluorescence quantification polymerase chain reaction (qRT-PCR). The results showed that the 50% effective concentration (EC₅₀) of CA against influenza H5N1 in A549 cells and MDCK cells were 4.30 and 3.64 μmol·L^-1, respectively. Meanwhile, CA also showed inhibition on influenza virus 2009panH1N1 (EC₅₀: 10.1 μmol·L^-1) and H3N2 (EC₅₀: 12.8 μmol·L^-1) replications in A549 cells. Mechanistic studies showed that antiviral activity of CA is related to its induction of heme oxygenase-1 (HO-1) in A549 cells and suppression on production of reactive oxygen in H5N1-infected cells.