Objective By constructing mortalin stably expressing ovarian cancer cell lines in A 2780 and A2780/cis, we demonstrate the role of mortalin in the ovarian cancer cell growth .Methods CCK-8 assay was used to measure cell viability in the overexpression mortalin group compared with the control group .The flow cytometry analysis was used to understand the effect of upregulated mortalin on the ovarian cancer cell cycle .Western blotting was used to determine the expression and phosphorylation level of MAPK /ERK and JNK/SAPK signal pathways .Results The results showed that increased expression of mortalin could accelerate ovarian cancer cell proliferation and promote G 1 transition, leading to a faster restoration of normal distribution of cell cycle .We found that mortalin overexpression significantly activated p-Raf and p-ERK1/2, but not p-JNK.Conclusion The results demonstrate that mortalin effect on the ovarian cancer cell proliferation contributes to active the MAPK-ERK signaling pathway .

Objective To study the effect of glucose regulated protein 75(Grp75) on the alteration of Bax and NF-κB induced by glucose deprivation through the stably transfected PC12 cells with Grp75. Methods The cells of Grp75-overexpressing group and control group incubated in glucose-free DMEM medium for indicated time (6, 12, 24 and 48hours). The expression level of Grp75, Bax and the activity of NF-κB were determined by Western blotting, and the expression level of Bax was determined by semi-quantitative RT-PCR and Western blotting. Immunocytochemistry was performed using a conformation specific anti-Bax (6A7) antibody to detect the activation of Bax. Results The activation of Bax and the decline of NF-κB activity played important roles in the apoptosis of PC12 cells induced by glucose deprivation. Grp75 inhibited the apoptosis induced by glucose deprivation through inhibition of the activation of Bax and the decline of NF-κB activity. There was no change in Bax expression level under glucose deprivation in two groups. Conclusion The activation of Bax and the decline of NF-κB activity were associated with apoptosis of PC12 cells induced by glucose deprivation, and Grp75 provided protection to PC12 cells through inhibition of activation of Bax and maintaining activation of NF-κB.

0.05).And the increasing of weight in high protein+plus prolein jelly group was significantly higher than those of other two groups(Pa

BACKGROUND:Endothelial progenitor cells are precursor cells of mature endothelial cells,which can migrate to ischemic tissues and differentiate into mature endothelial cells,and then play an important role in vascular remodeling.Endothelial progenitor cells have wide application prospects in various ischemic diseases,but the biological characteristics and identification methods are still controversial.OBJECTIVE:To investigate the methods of isolation and culture of endothelial progenitor cells from the human adipose tissue and to identify their biological features,in order to provide a sufficient source of cells for ischemic diseases.METHODS:Stromal vascular fraction cells were isolated from the human adipose tissue by enzymatic digestion,CD31+ cells were selected using immunomagnetic beads,and then cultured in endothelial basal medium-2 supplemented with the EGM-2-MV-SingleQuots.Endothelial progenitor cells were identified through detection of morphology,cell markers and cell functions.RESULTS AND CONCLUSION:(1) CD31 + cells were selected by immunomagnetic beads and then cultured and amplified in vitro,which displayed typical cobblestone-like morphology,and they maintain their proliferative ability.(2) Flow cytometry results showed that the CD31+ cells expressed CD31 (98.84％),CD34 (97.21％),VEGRR2 (64.07％),CD146 (98.42％) and CD133 (2.55％),but hardly expressed CD45 (1.1％),a hematopoietic stem cell marker.(3) The CD31 + cells were also found to incept Dil-ac-LDL and exhibit lectin binding capability.Furthermore,a lumen-like structure was formed in Matrigel,which has the ability of angiogenesis in vitro.To conclude,these results suggest that it is feasible to isolate and culture endothelial progenitor cells from the human adipose tissue by enzymatic digestion combined with immunomagnetic bead sorting.

OBJECTIVETo investigate the protective effect of lycopene against cryopreservation injury of post-thawing human sperm and its mechanism.

METHODSSemen samples were collected from 25 volunteers, each sample equally divided into four parts to be cryopreserved with cryoprotectant only (Ly0 control) or cryoprotectant + lycopene at the concentrations of 2 (Ly2), 5 (Ly5), and 10 µmol/L (Ly10), respectively. Before and after thawing, the semen samples were subjected to computer-assisted semen analysis ( CASA) for sperm kinematics, flow cytometry for sperm apoptosis, thiobarbituric acid assay for malondialdehyde (MDA) concentration, and JC-1 fluorescent staining for the sperm mitochondrial membrane potential (MMP).

RESULTSAfter cryopreservation, sperm motility was markedly decreased in all the groups (P < 0.01). The rate of sperm apoptosis was significantly lower in the Ly5 group than in the Ly0 control ([25.68 ± 4.36]% vs [33.26 ± 4.78]%, P < 0.05), while sperm MMP remarkably higher in the former than in the latter ([66.18 ± 14.23]% vs [55.24 ± 12.31]%, P < 0.05). The Ly2, Ly5 and Ly10 groups showed no statistically significance differences in the MDA level from the Ly0 control (P > 0.05).

CONCLUSIONAddition of lycopene at a proper concentration to cryoprotectant may reduce oxidative damage to sperm mitochondria in the freezing-thawing process, attenuate oxidative stress injury induced by reactive oxygen species to sperm plasma membrane, and improve the anti-apoptosis ability of sperm.

Apoptosis ; Carotenoids ; pharmacology ; Cryopreservation ; Cryoprotective Agents ; pharmacology ; Flow Cytometry ; Humans ; Male ; Malondialdehyde ; analysis ; Oxidative Stress ; Reactive Oxygen Species ; Semen Analysis ; Semen Preservation ; adverse effects ; methods ; Sperm Motility ; Spermatozoa ; drug effects ; physiology

OBJECTIVETo prove if it is possible for using the shattering extraction with solvent to extract ingredients of traditional Chinese medicine.

METHODThe shattering extraction with solvent, the refluxing extraction and the ultrasonic extraction were used to extract paeoniflorin from Radix Paeoniae rubra, and to extract baicalein from Radix Scutellariae, and to extract chlorogenic acid from Flos lonicerae japonicae respectively, using ingredient content and extract yield as the measuring indexes.

RESULTThe content of each every ingredient obviously higher by using shattering extraction with solvent than using refluxing extraction or the ultrasonic extraction.

CONCLUSIONThe shattering extraction with solvent is a high efficiency, simple and quick extraction. It may be used to extract the ingredient of three kinds of traditional Chinese medicine.

Benzoates ; isolation & purification ; Bridged-Ring Compounds ; isolation & purification ; Chemical Fractionation ; methods ; Chlorogenic Acid ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; isolation & purification ; Glucosides ; isolation & purification ; Monoterpenes ; Solvents ; chemistry ; Time Factors

Objective To illustrate if the racial coefficient (Rc) be biased by different reference GFR (rGFR) distribution among studies. Methods 1405 white and 321 African American participants in MDRD study and 684 chronic kidney disease(CKD)patients in Chinese eGFR Investigation Study were included.Firstly.the unweighted datasets of white and Chinese were stacked together.rGFR,age and plasma creatinine (Pcr) were log transformed.Linear regression model was constructed using log transformed rGFR as dependent,gender,race and log transformed Pcr and age as independent.Unweighted RC (uRC) for Chinese was calculated.Then.the Chinese CKD distribution of rGFR was weighted to be the same as that in White American.and weighted RC (wRC)for Chinese was calculated.The cases of White.and African-American were stacked together.The cases of African-American were weighted to make the rGFR distribution the same as that in White-American and Chinese population respectively,and RCs for African-American were calculated. Resuits The uRC for Chinese was 1.197(1.180-1.211)and the wRC was 1.130 (1.117-1.143).The two RCs did not overlap with each other.The RCs for African-American were 1.205(1.19-1.219)and 1.233(1.219-1.247)respectively. Conclusions The RCs were influenced by the difference of rGFR distribution.To find out the real RC.an intemational collaborative study iS needed,with the same rGFR measure method.strict control of Pcr measurement,and the same rGFR distribution.

OBJECTIVETo research (CAG)n polymorphism of androgen receptor gene in idiopathic azoospermic and oligospermic Chinese men, and to explore its correlation with the development of dyszoospermia.

METHODSFifty-two men with oligospermia and 31 with azoospermia were enrolled, the number of CAG sequence repeats determined by PCR and denaturing polyacrylamide gel electrophoresis (PAGE), and (CAG)n polymorphism analyzed.

RESULTSThe CAG repeats in the men with oligospermia and those with azoospermia were 22.19 and 22.13 respectively.

CONCLUSIONThe polymorphism of the repeat length of the (CAG) n satellite in androgen receptor gene may affect spermatogenesis and have a less important role in male infertility.

Adult ; Electrophoresis, Polyacrylamide Gel ; Exons ; Humans ; Infertility, Male ; genetics ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Receptors, Androgen ; genetics ; Trinucleotide Repeats ; genetics

Adolescent ; Adult ; Child ; Child, Preschool ; Electroencephalography ; Epilepsy ; pathology ; physiopathology ; Female ; Hippocampus ; pathology ; Humans ; Male

BACKGROUND: Adipose-derived stem cells (ADSCs) can establish a favorable repair microenvironment by secreting abundant cytokines, which allows ADSCs to be a good source of seed cells for the treatment of ischemic diseases. OBJECTIVE: To investigate the changes of cytokines secreted by human ADSCs at passages 2-10. METHODS: After isolation and culture of ADSCs from human adipose tissue, the morphological features of cells were observed under inverted microscope. Human ADSCs were identified by the immunophenotypes and differentiation capability. RESULTS AND CONCLUSION: ADSCs were fusiform or polygonal in shape, with buging cell body, homogenized cytoplasm and clear nuclei, and could differentiate into adipocytes, osteocytes and chondroblasts in vitro. ADSCs at passage 3 were positive for CD29 (99.21%), CD73 (99.65%) and CD90 (99.92%), but negative for hematopoietic marker CD34 (2.25%). ELISA results showed that ADSCs at passage 5 had the highest secretion levels of vascular endothelial growth factor and hepatocyte growth factor, while ADSCs at passage 3 had the highest secretion level of brain-derived neurotrophic factor. To conclude, ADSCs have steady biological features of stem cells and exhibit good growth and proliferation potentials. ADSCs at different passages have different capacities in the secretion of vascular endothelial growth factor, hepatocyte growth factor and brain-derived neurotrophic factor. Passage 5 ADSCs show the highest ability to secrete vascular endothelial growth factor and hepatocyte growth factor, while passage 3 ADSCs show the strongest potential to secrete brain-derived neurotrophic factor.