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

To study the effects and possible mechanism of Vitamin K(2) (VK(2)) in the treatment of MDS-JSN04 cells, the changes of morphologic features of MDS-JSN04 cells were investigated by cytomorphology, the apoptosis of MDS-JSN04 cells was observed by transmission electron microscope; cellular proliferation was determined by the MTT assay; cell apoptosis, cell cycle shift and expression of myeloid-specific differentiation antigen (CD11b, CD13) were analyzed by flow cytometry (FCM). The expression of apoptosis-related genes bcl-2, survivin and bax were detected by retrotranscriptase polymerase chain reaction (RT-PCR); the activity of caspase-3 was determined by chemiluminescence assay. The results showed that the typical apoptotic morphological features appeared in cells treated with VK(2) for 72 hours; VK(2) induced apoptosis of MDS-JSN04 cells and in a dose-and-time-dependent manner, G(0)/G(1) cell arrest and significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax; the activity of caspase-3 significantly increased. It is concluded that VK(2) induces apoptosis of MDS-JSN04 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2, survivin may play an important role in this process.
Apoptosis ; drug effects ; CD11b Antigen ; analysis ; CD13 Antigens ; analysis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Luminescent Measurements ; methods ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; genetics ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Vitamin K 2 ; pharmacology ; bcl-2-Associated X Protein ; genetics

OBJECTIVETo find the effects of lead taken by pregnant mice on learning and memory and the expression of synaptosomal-associated protein (SNAP)-25 mRNA and protein, in order to reveal the mechanism of neurotoxicity induced by lead.

METHODSLead exposure was conducted through freely drinking the corresponding lead acetate solutions with dosages of 0.3, 1.0, 3.0 g/L respectively. Each group was composed of 10 mice. 7, 14 and 21 days after their birth. The lead contents in blood and hippocampus of the offspring were determined. At the 21st day the expression of SNAP-25 mRNA and protein in hippocampus of all the offspring in various dosages groups were determined by RT-PCR and immunohistochemistry assay.

RESULTSThe lead contents in blood and hippocampus of various lead exposed groups were significantly higher than those of the control group (P < 0.05). The lead levels in blood and hippocampus changed accordingly to the days of growth. In Water Morris Maze experiment, the result of 0.3 g/L group was not significantly different from that of the control group (P > 0.05), however, the results of 1.0, 3.0 g/L groups (5.89 ± 0.54, 9.53 ± 1.03) were significantly different from those of the control group (1.73 ± 0.07) (P < 0.05, P < 0.01). The expression of SNAP-25 mRNA and protein was lower in lead exposed groups than that of the control group (P < 0.05).

CONCLUSIONMaternal lead exposure may induce the damage in the ability of learning and memory of the offspring. The neurotoxicity of lead may be induced by decreasing the expression of SNAP-25 mRNA and protein so as to affect the release of neurotransmitter from presynaptic terminal resulted in nerve damages.

Animals ; Female ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Maternal Exposure ; Maze Learning ; drug effects ; Memory ; drug effects ; Mice ; Pregnancy ; RNA, Messenger ; genetics ; Synaptosomal-Associated Protein 25 ; 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