This study was aimed to investigate the effects and the mechanism of mangiferin on chronic myeloid leukemia cell lines K562 cells in vitro. The antiproliferation effects of mangiferin on K562 leukemia cells were tested by tetrazolium salt (MTT) method; the apoptosis induced by mangiferin on K562 cell line was explored by means of cell morphology, DNA gel electrophoresis and flow cytometry. The changes in bcr/abl gene expression was detected by using reverse transcriptase (RT)-PCR. The results showed that five different concentrations of mangiferin (25 - 200 micromol/L) dose-dependently and time-dependently inhibited the proliferation of K562 cells, and induced apoptosis in K562 cell line. RT-PCR revealed that bcr/abl gene expression was down-regulated when K562 cells had been treated with different concentrations of mangiferin. In conclusion, mangiferin remarkably inhibits the proliferation of K562 leukemia cells in vitro, and induces apoptosis in K562 cell line probably through down-regulation of bcr/abl gene expression.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; DNA Fragmentation ; drug effects ; Flow Cytometry ; Genes, abl ; Humans ; K562 Cells ; Xanthones ; pharmacology

OBJECTIVETo investigate the role of Annexin 5 in protecting human sperm membrane and DNA integrity.

METHODSWe collected 53 semen samples based on the criteria of sperm density > 20 x 10(6)/ml and motility > 60%, and divided them into an experimental group (2.5 microl 10(-6) mol/L Annexin 5 added to 47.5 microl semen), a negative control group (2.5 microl 1 mol/L Tris-HCl [pH 8.0, 25 degrees C] added to 47.5 microl semen), and a blank control group (2.5 microl 0.01 mol/L PBS [pH 7.4] added to 47.5 microl semen). After 20 minutes of incubation, we evaluated the sperm membrane integrity using the hypoosmotic swelling test and, after another 60 minutes of treatment with H2O2 at 2.5 microl 10.02 mol/L, measured the sperm nuclear DNA integrity by acridine orange fluorescent staining.

RESULTSAfter 20 minutes of treatment with Annexin 5, the experimental group showed extremely significant difference in the percentage of hypoosmotic swelling sperm ([66.17 +/- 12.02] %) from the blank control ([58.13 +/- 13.08]%, P < 0.01) and the negative control group ([59.94 +/- 11.91]%, P < 0.01), but there was no significant difference between the latter two. Treatment with H2O2 remarkably increased DFI in the experimental group (6.39 +/- 1.07) as compared with the blank control (11.16 +/- 1.16) and the negative control group (10.86 +/- 1.05, P < 0.01), but no significant difference was observed between the latter two.

CONCLUSIONAnnexin 5 can increase the percentage of hypoosmotic swelling sperm in vitro and protect sperm membrane integrity, and it can also protect sperm DNA from H2O2 damage.

Annexin A5 ; pharmacology ; Cell Membrane ; drug effects ; DNA ; DNA Fragmentation ; Humans ; Male ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects

OBJECTIVETo observe calf thymus DNA damage induced by potassium dichromate in combination with glutathione (GSH).

METHODSAtomic force microscope and ultraviolet spectrum (UV) were used to observe the alterations of the DNA ultrastructure and absorption spectrum.

RESULTSAtomic force microscopy revealed no breaks of the DNA strand in response to treatment with potassium dichromate alone, but when coupled with GSH at proper concentrations, potassium dichromate induced alterations in the DNA structure and DNA fragmentation. UV examination also confirmed these findings by showing increased absorption intensity of the maximum UV peak following combined treatment of the DNA with potassium dichromate and GSH.

CONCLUSIONThese morphological and spectrographic evidences verified the important role of GSH in mediating the generation of various tumor-inducing intermediate products of potassium dichromate.

Animals ; Cattle ; DNA ; chemistry ; genetics ; DNA Damage ; DNA Fragmentation ; drug effects ; Glutathione ; toxicity ; Microscopy, Atomic Force ; methods ; Nucleic Acid Conformation ; drug effects ; Potassium Dichromate ; toxicity ; Spectrophotometry, Ultraviolet

OBJECTIVETo investigate the influence of cigarette smoking on human sperm DNA integrity.

METHODSTotally, 784 cases of male infertility were selected from our case database and grouped according to whether they were smokers or nonsmokers, how much they smoked (< or = 10, 11-19 and > or = 20 cigarettes/d) and how long they smoked (< or = 5, 6-9 and > or = 10 yr). Sperm DNA integrity was measured using sperm chromatin structure assay (SCSA) and flow cytometry. DNA fragmentation and immature spermatozoa were expressed by the DNA fragmentation index (DFI) and high DNA stainability (HDS) respectively. Conventional sperm parameters and sperm DNA integrity were compared among different groups.

RESULTSThe total semen volume and percentage of grade a + b sperm were lower and the sperm morphological abnormality was higher in the > or = 20 cigarettes/d and > or = 10 yr groups than in the others (P < 0.05). DFI and HDS were significantly higher in the smokers than in the nonsmokers (P < 0.05). HDS was negatively correlated with the percentage of grade a + b sperm (r = -0.18, P < 0.05) and both DFI and HDS were positively correlated with the rate of sperm malformation (r = 0.31 and r = 0.39, P < 0.05).

CONCLUSIONSmoking more than 20 cigarettes a day or longer than 10 years has deleterious effects on the semen volume, percentage of grade a + b sperm and sperm morphology of the smokers. Cigarette smoking decreases sperm DNA integrity and nuclear maturation.

Adult ; DNA Damage ; drug effects ; DNA Fragmentation ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Smoking ; adverse effects ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; Young Adult

In order to study the influence of trichosanthin (TCS) on apoptosis and growth inhibition of human NB4 cells in vitro, the expression of annexin V and the change of DeltaPsim of NB4 cells induced by TCS was analyzed by FACS, and MTT assay was adopted to measure the growth inhibition ratio of NB4 cells treated with TCS. Apoptosis was assayed by agarose gel electrophoresis. The results showed the higher concentration of TCS and the longer the acting time, the stronger growth inhibition of NB4 cells. The expression of annexin V was positive, and the positive ratio was greatly enhanced with prolongation of acting time. DeltaPsim reduced gradually while the apoptosis cells increasing. DNA agarose gel electrophoresis showed a gradient, which confirmed that TCS could induce NB4 cells apoptosis. In conclusion, taken together, data show that TCS can inhibit NB4 growth in vitro, and induce apoptosis. Experiment provides an important evidence for application of TCS in clinical treatment of acute promyelocytic leukemia.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Fragmentation ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Trichosanthin ; pharmacology

Objective: To clarify the roles of yam polysaccharide (YPS) in improving sperm viability and protecting sperm DNA integrity in vitro and provide a new approach to the treatment of oligoasthenozoospermia. METHODS: We collected samples by masturbation from 36 normal fertile males aged 27－39 years. Each sample was divided into six groups: blank control or treated with normal saline, vitamin C solution, and YPS solution at low (0.25 mg/ml), medium (1.0 mg/ml) or high concentration (5.0 mg/ml). Using eosin-Y staining, sperm hypotonic swelling (HOS) and sperm chromatin diffusion (SCD) test, we observed the effects of different concentrations of YPS on sperm viability, membrane integrity and nuclear DNA. RESULTS: After 24 and 48 hours of treatment, sperm viability was markedly reduced in the vitamin C (［28.5 ± 3.1］ and ［6.5 ± 1.2］%), low-YPS (［31.3 ± 3.5］ and ［6.5 ± 2.2］%), medium-YPS (［37.1 ± 3.5］ and ［9.5 ± 2.8］%) and high-YPS groups (［38.3 ± 3.3］ and ［9.0 ± 3.2］%) as compared with the blank control (［17.3 ± 2.1］ and ［3.2 ± 1.3］%) (P <0.01) and normal saline groups (［13.4 ± 4.1］ and ［3.1 ± 2.0］%) (P <0.01), and it was significantly higher in the medium- and high-YPS than in the vitamin C group (P <0.05 and P <0.01). The rate of sperm DNA fragmentation was remarkably decreased at 48 hours in the vitamin C (［30.5 ± 3.1］%), low-YPS (［29.4 ± 2.6］%), medium-YPS (［28.5 ± 2.3］%) and high-YPS groups (［27.9 ± 1.9］%) in comparison with the blank control (［41.7 ± 2.2］%) (P <0.01) and normal saline groups (［42.1 ± 3.3］%), markedly lower in the medium- and high-YPS than in the blank control, normal saline and vitamin C groups (P <0.05 or P <0.01), but with no statistically significant difference between the low-YPS and vitamin C groups (P >0.05). CONCLUSIONS Yam polysaccharide can improve sperm viability and protect sperm DNA integrity in vitro.
Adult ; Ascorbic Acid ; pharmacology ; DNA ; drug effects ; DNA Fragmentation ; Dioscorea ; chemistry ; Humans ; Male ; Polysaccharides ; pharmacology ; Semen Analysis ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Vitamins ; pharmacology

Treatment with certain DNA-damaging agents induce a complex cellular response comprising pertubation of cell cycle progression and/or apoptosis on proliferating mammalian cells. Our studies were focused on the cellular effects of nickel (II) acetate, DNA-damaging agent, on Chinese hamster ovary (CHO) cells. Fragmented DNAs were examined by agarose gel electrophoresis and cell cycle was determined by DNA flow cytometry using propidium iodide fluorescence. Apparent DNA laddering was observed in cells treated with 240 microM nickel (II) and increased with a concentration-dependent manner. Treatment of nickel (II) acetate resulted in apoptosis which was accompanied by G2/M cell accumulation. Proportion of CHO cells in G2/M phase was also significantly increased in cells exposed to at least 480 microM nickel (II) from 57.7% of cells in the G0/G1 phase, 34.7% in the S phase, and 7.6% in the G2/M1 phase for 0 microM nickel (II), to 58.6%, 14.5%, and 26.9% for 640 microM nickel (II). These findings suggest that nickel (II) can modulate cellular response through some common effectors involving in both apoptotic and cell cycle regulatory pathways.
Animal ; Apoptosis/drug effects* ; CHO Cells/drug effects* ; CHO Cells/cytology ; Cell Cycle/drug effects* ; DNA Fragmentation/drug effects ; Flow Cytometry ; G2 Phase/drug effects ; Hamsters ; Mitosis/drug effects ; Nickel/pharmacology*

AIMTo observe the cytotoxic effect of the organophosphate insecticide malathion in the reproductive tissues of the earthworms, Eisenia foetida.

METHODSWorms were nourished in soil treated with malathion at single sub-lethal doses of 0, 80, 150, 300 and 600 mg/kg soil. (LD50=880 mg/kg soil) and evaluated on days 1, 5, 15 and 30 after exposure. The body weights were recorded and male reproductive organs evaluated.

RESULTSMalathion-treated animals showed a significant reduction in body weight in a dose-dependent manner. Malathion treatment modified the disposition of spermatozoa in the basal epithelium of the spermatheca. The Br-deoxyuridine test showed a significant rise in cells in phase S on days 5 and 15. Also, a higher percentage of spermatogonia with fragmented DNA were observed by means of the TdT-mediated dUTP nick-end labeling (TUNEL) technique in the spermatheca of treated animals.

CONCLUSIONTreatment with malathion decreased the body weight and the spermatic viability in spermatheca, altering the cell proliferation and modifying the DNA structure of spermatogonia.

Animals ; Body Weight ; drug effects ; DNA Fragmentation ; Dose-Response Relationship, Drug ; In Situ Nick-End Labeling ; Malathion ; adverse effects ; Male ; Oligochaeta ; drug effects ; Reproduction ; drug effects ; S Phase ; drug effects ; genetics ; Spermatozoa ; drug effects ; Time Factors

OBJECTIVETo investigate the clinical effects of the combined therapy of the Chinese medicine Compound Xuanju Capsule and vitamin E on sperm chromatin damage in idiopathic oligoasthenospermia.

METHODSWe assigned 50 infertile men with seminal abnormality to a control group (n = 26) and a trial group (n = 24) to receive vitamin E and the combined therapy of Compound Xuanju Capsule plus vitamin E, respectively, both treated for 3 months. Before and after the treatment, we detected semen routine parameters and sperm DNA fragmentation indexes (DFI) by computer aided semen analysis (CASA) and sperm chromatin structure assay (SCSA), and compared them between the two groups.

RESULTSThere was no obvious difference between the percentage of progressively motile sperm in the trial group and that in the control group (21.55 +/- 8.68 vs 21.47 +/- 11.53, P > 0.05). The trial group showed a significantly decreased sperm DFI after medication as compared with pre-medication (29.57 +/- 12.19 vs 34.09 +/- 10.32, P < 0.05).

CONCLUSIONThe combined therapy of Compound Xuanju Capsule and vitamin E can effectively improve seminal quality and reduce sperm chromatin damage in infertile men with idiopathic oligoasthenospermia.

Adult ; Capsules ; Chromatin ; drug effects ; DNA Damage ; drug effects ; DNA Fragmentation ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; genetics ; Male ; Spermatozoa ; drug effects ; Vitamin E ; pharmacology ; therapeutic use ; Young Adult

A water-soluble polysaccharide fraction from root of Potentilla anserine was obtained. Gas chromatogram, FT-IR, physical and chemical characteristics of the Potentilla anserine polysaccharide fraction (PAPF) were analyzed. The protective effects of PAPF against the H2O2 induced process of apoptosis of murine splenic lymphocytes were investigated in vitro. Morphological assessment of apoptosis was performed with light microscope and laser scanning confocal microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. The results showed that PAPF is composed of rhamnose, arabinose glucose and galactose. H2O2 (200 micromol x L(-1)) induced apoptosis of murine splenic lymphocytes with the cell volume reduced, cytoplasm and nuclear shrunk and DNA stained non-uniformly. Condensed chromatin and formation of apoptotic body were observed in the apoptotic cells. Apoptotic bodies in the cells treated with PAPF and H2O2 were less than those in H2O2 treatment alone. DNA fragmentation assay showed that PAPF (50, 100, 200, and 400 microg x mL(-1)) obviously reduced H2O2-induced ladder bands. Flow cytometry analysis showed that H2O2 increased the populations of apoptotic sub-G1 cells from 5.60% (control) to 45.40%, and PAPF decreased H2O2-induced apoptosis to 37.80%, 22.70%, 17.70%, and 8.50%, respectively. In conclusion, PAPF reduced H2O2-induced oxidative damage in a dose dependent manner.
Animals ; Apoptosis ; drug effects ; Cells, Cultured ; DNA Fragmentation ; Flow Cytometry ; Hydrogen Peroxide ; pharmacology ; Lymphocyte Count ; Lymphocytes ; cytology ; drug effects ; Mice ; Polysaccharides ; pharmacology ; Potentilla ; Spleen ; cytology