Animals ; Diet, High-Fat ; Glucagon ; blood ; Glucose ; metabolism ; Hormones ; blood ; Insulin ; blood ; Leptin ; blood ; Lipid Metabolism ; Male ; Rats ; Rats, Sprague-Dawley

The aim of this study was to investigate the effects of hypoxia conditioned medium (HCM) of cerebral cortex cells on the differentiation of neural stem cells (NSCs) and to clarify the signal transduction mechanism. The cerebral cortex cells from newborn SD rats were primarily cultured for 5 d, and then the cells were cultured in environments of 4% O2, 1% O2 and normal oxygen concentration, respectively, for 6 h. The culture mediums were collected and centrifuged as the HCM and normoxia conditioned medium (NCM). The neurospheres of NSCs were obtained from the rat cerebral cortex by suspending culture. Immunohistochemical staining was used after adherence culture for 48 h to identify neurons and astrocytes in the progeny of NSCs. LY294002, a PI3-K inhibitor, and SP600125, a JNK inhibitor, were added into the HCM to culture NSCs for 48 h. The results showed that NSCs in the cerebral cortex could differentiate into β-Tubulin III immunoreactive neurons and GFAP immunoreactive astrocytes in three conditioned mediums, and the neurons proportion in progeny of NSCs was higher than astrocytes in all three groups. The proportion of neurons in 4% HCM was higher than that in NCM (P < 0.01). But the proportion of neurons in 1% HCM was less than that in NCM (P < 0.01). Both LY294002 and SP600125 inhibited NSCs to differentiate into high proportion neurons induced by 4% HCM (P < 0.01), but the inhibitory effect of LY294002 was stronger than that of SP600125 (P < 0.01). In conclusion, 4% HCM can induce NSCs to differentiate into more neurons mainly through the PI3-K pathway.
Animals ; Animals, Newborn ; Astrocytes ; cytology ; Cell Culture Techniques ; Cell Differentiation ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; cytology ; Culture Media, Conditioned ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

To investigate the effect of treadmill running on the ability of learning in young rats, male Sprague-Dawley rats (5 weeks of age) were used for the experiment. Animals were randomly divided into the control and running groups (n=15 in each group). The rats in running group were made run on a motor-driven treadmill for 1 week at a speed of 2 m/min for the first 5 min, at a speed of 5 m/min for the next 5 min, then at a speed of 8 m/min for the last 20 min. Then the Morris water maze was used to observe learning and memory ability of rats in both groups. The tests consisted of place navigation and spatial probe test. We found that, in place navigation training, the latency of rats in running group was less than that in control group (P<0.05); and from the third training session on, there was significant difference between the rats in control and running groups in swimming velocity (P<0.01); furthermore, it was observed that the rats in running group had stronger motivation and more exact orientation in searching for platform, which could be indicated by the index of turn angle and angular velocity. In spatial probe test, there was no significant difference between the two groups in swimming velocity, percentage of swimming distance and frequency of crossing platform in D quadrant, where the platform situated (P>0.05). These findings suggest that low speed treadmill running can enhance the ability of learning in young rats.
Age Factors ; Animals ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Physical Conditioning, Animal ; physiology ; Rats ; Rats, Sprague-Dawley ; Spatial Behavior ; physiology ; Swimming ; physiology

AIMTo analyse the mechanism of corticosterone on the elevation of cytosolic free calcium ([Ca2+]i) induced by high-K+ in pheochromocytoma PC12 cells,

METHODSThe [Ca2+]i was real-time checked by fluorescence image system.

RESULTS(1) When the cells were preincubated at 37 degrees C for 5 min in the presence of various concentration corticosterone and stimulated with 55 mmol/L KCl , an inhibitory effect of corticosterone on delta[Ca2+]i was observed in a concentration-dependent manner. (2) When PC12 cells were preincubated with various concentration of B-BSA at 37 degrees C for 5 min and stimulated with 55 mmol/L KCl, an inhibitory effect of B-BSA on delta[Ca2+]i was observed, which is also concentration-dependent manner. (3) The inhibitory effect of corticosterone and B-BSA could not be antagonized by RU38486 at 10(-4) mol/L. (4) cycloheximide could not block the inhibitory effect of corticosterone after pretreating cells at 10(-5) mol/L at 37 degrees C for 3 hours.

CONCLUSIONIt is obvious that the locus of corticosterone action is on the plasmic membrane. The inhibitory effect of corticosterone is independent of protein synthesis and intracellular glucocorticoid receptor. The effect of corticosterone on [Ca2+]i is nongenomic action in PC12 cells.

Animals ; Calcium ; metabolism ; Corticosterone ; pharmacology ; PC12 Cells ; drug effects ; Potassium ; pharmacology ; Rats

AIMTo explore the effects of noninvasive limb ischemic preconditioning on the anti-stress ability in mice.

METHODSMice were divided into: normal group, control group, preconditioning group and drug group. Hypoxia tolerance test, swimming with weight loading, cold tolerance test and thermostable test were performed, and tolerance time in all the stringent state were observed. SOD activity of serum in hypoxia tolerance test and lactic acid of serum in swimming with weight loading test were determined.

RESULTSThe time of hypoxia tolerance in preconditioning group was markedly increased, and SOD activity of preconditioning group mice was significantly higher than those of control group, while they were both shorter than drug group. The average time of swimming in preconditioning group was markedly increased and the level of increasing the swimming time of preconditioning was the same as caffeine. Preconditioning could increase the survival time on high temperature markedly, and there was no significantly difference in the level of increasing the survival time between preconditioning group and chlorpromazine group. Preconditioning could increase the time of cold tolerance markedly compared with normal group.

CONCLUSIONNoninvasive limb ischemic preconditioning can improve the ability of anti-hypoxia, anti-fatigue, thermoresistance and cold-resistance in mice.

Adaptation, Physiological ; physiology ; Animals ; Extremities ; blood supply ; Fatigue ; prevention & control ; Female ; Hypoxia ; prevention & control ; Ischemic Preconditioning ; methods ; Male ; Mice ; Stress, Physiological ; physiology

ObjectiveTo observe the morphological changes of bone in the progress of chronic fluorosis.MethodsWistar rats were randomly divided into three groups,30 rats in each group:normal control group,experimental group Ⅰ and experimental group Ⅱ according to body weight.Rats in normal control group drank distilled water freely.Experimental group Ⅰ and group Ⅱ drunk distilled water with sodium fluoride preparation of fluorine containing ion 100,150 mg/L solution for six months,respectively.Bone mineral density was detected by X-ray,bone morphological changes were observed under light microscope and bone histomorphometric parameters were calculated using image analysis software.ResultsThe bone mineral density values were different statistically between the three groups after feeding for 2 and 4 months(F =19.79,3.28,all P ＜ 0.05).However no significant difference was found after feeding for 6 months(F =1.80,P ＞ 0.05).The bone mineral density of experimental group Ⅰ (0.20 ± 0.03,0.21 ± 0.03) was significantly higher than that of the normal control group(0.17 ± 0.03,0.20 ± 0.04) after feeding for 2 and 4 months.The bone mineral density of experimental group Ⅱ (0.21 ± 0.02) was lower than that of normal control group(0.22 ± 0.03) after feeding for 6 months.The bone lamella in experimental group Ⅰ was arranged disorderly,the number of osteocytes increased with their nucleus atrophy and the osteoblasts were more than that of control grouo which arranged in layers observed under light microscooy.In exoerimental group Ⅱ,the bone lamella was bent deformation,the number of osteocytes had decreased with their nucleus shrinking or even disappeared and the number of osteoclasts had increased significantly observed under light microscopy.In experimental group Ⅰ,the mean trabecular density [(0.33 ± 0.03)％] increased and the mean trabecular separation,thickness [( 163.57 ± 1.99),(59.26 ± 7.18 ) μm] decreased compared with that of normal control group [(0.31 ± 0.02)％,(186.60 ± 2.90)μm,(86.42 ± 1.48)μm,all P ＜ 0.05].In experimental group Ⅱ,the mean trabecular density[(0.26 ± 0.02)％] decreased,the mean trabecular thickness[(71.42 ± 10.77)μm] reduced compared with that of normal control group[(0.31 ± 0.02)％,(86.42 ± 1.48)μm].ConclusionsExcess fluoride can damage bone tissue.Low doses of fluoride can stimulate osteoblast activity and enhance osteogenesis.The activity of osteoblasts is great than that of osteoclasts.High doses of fluoride can stimulate both osteoblasts and osteoclasts activity,but mainly the activity of osteoclasts,and bone resorption increases.

ObjectiveTo detect the concentration and distribution of fluoride ions in osteoblasts exposed to fluoride in vitro culture,and to provide basic information for studying the effect of fluoride on osteoblast injury.MethodsIn vitro cultured osteoblasts were exposed to 0,5,10,20,40 mg/L fluoride for 3,10,30 d (n =6),respectively.Concentration and distribution of fluoride ions in the cytoplasm and the nucleus of these osteoblasts were determined by nuclear magnetic resonance spectroscopy.Results(①) After cultured for 3 d,fluoride ion content of the bone cytoplasm exposed to different concentrations of fluoride 0,5,10,20,40 mg/L were (0.83 ±0.65),(0.54 ± 0.23),(0.65 ± 0.77),(0.59 ± 0.87),(3.64 ± 1.21 )mg/L,respectively,and the values of exposed to 40 mg/L fluoride group was significantly higher than that of exposed to 0,5 mg/L groups (all P ＜ 0.05).(②)after cultured for 10 d,the composition of the fluoride ion in cytoplasm of exposed to fluoride 10,20,40 mg/L groups were (4.03 ± 1.23),(3.66 ± 0.98),(6.26 ± 2.10)mg/L,respectively,which were higher than that of exposed to 0,5 mg/L groups [(0.78 ± 0.75),(2.69 ± 0.89)mg/L,respectively,all P ＜ 0.05].Of fluoride 20,40 mg/L groups,the composition of the fluoride ion in nucleus were (1.63 ± 1.19),(2.17 ± 1.21 )mg/L,respectively,which were higher than that of 0,5 mg/L groups[(0.65 ± 0.46),(1.57 ± 0.33) mg/L,all P ＜ 0.05].(③)After cultured for 30 d,of the exposed to fluoride 10,20,40 mg/L groups,the composition of the fluoride ion in cytoplasm were (3.99 ± 0.84),(4.33 ± 1.67),(5.80 ± 1.38)mg/L,respectively,which were higher than that of 0,5 mg/L groups[(0.88 ± 0.44),(2.84 ± 0.43)mg/L,all P ＜ 0.05].The composition of the fluoride ion in nucleus of the fluoride 20,40 mg/Lgroups were (3.33 ± 1.46),(3.53 ± 1.22)mg/L,respectively,which were significantly higher than that of 0,5mg/L groups [(0.70 ± 0.66),(1.99 ± 0.76)mg/L,all P ＜ 0.05].ConclusionsWhen osteoblasts are exposed to fluoride environment,fluoride ions enter into the osteoblasts quickly,and quickly accumulate in the nucleus,showing a special affinity between fluoride and bone tissue.Intracellular fluoride ions increase with the increase of contact time and exposure dose.

OBJECTIVETo investigate whether chronic bacterial prostatitis (CBP) increases oxidative stress and damage in patients with CBP, and to explore its possible mechanism.

METHODSEighty patients with CBP and 80 healthy adults as controls were enrolled in a case-control study, in which levels of nitric oxide (NO), vitamin C (VC), and vitamin E (VE) in plasma, as well as malondialdehyde (MDA), activities of superoxide dismutase (SOD), and catalase (CAT) in erythrocytes were determined by spectrophotometry.

RESULTSCompared with the average values of NO, VC, VE, MDA, SOD, and CAT in the healthy control group, those of plasma NO and erythrocyte MDA in the CBP group were significantly increased (P < 0.001), and those of plasma VC and VE as well as erythrocyte SOD and CAT in the CBP group were significantly decreased (P < 0.001). Findings from partial correlation analysis for course of the disease and NO, VC, VE, MDA, SOD, and CAT in 80 patients with CBP, adjusted for age, suggested that with prolonged course of the disease, values of NO and MDA were gradually increased (P < 0.001), and those of VC, VE, SOD, and CAT were gradually decreased (P < 0.05-0.001). The findings from stepwise regression analysis for course of the disease and NO, VC, VE, MDA, SOD, and CAT in CBP group suggested that the model of stepwise regression was Y = -19.1160 + 0.3112MDA + 0.0337NO, F = 22.1734, P < 0.001, r = 0.6045, P < 0.001. The findings from the reliability analysis for VC, VE, SOD, CAT, NO, and MDA in the CBP group showed that the reliability coefficients' alpha (6 items) was 0.7195, P < 0.0001, and the standardized item alpha was 0.9307, P < 0.0001.

CONCLUSIONThere exist increased oxidative stress and damage induced by chronic bacterial prostatitis in patients, and such a phenomenon is closely related to the course of disease.

Adult ; Ascorbic Acid ; blood ; Case-Control Studies ; Catalase ; metabolism ; Erythrocytes ; enzymology ; Humans ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; blood ; Oxidative Stress ; Prostatitis ; blood ; diagnosis ; Spectrophotometry ; Superoxide Dismutase ; metabolism ; Vitamin E ; blood

OBJECTIVETo evaluate the possibility of Id4 gene promoter methylation as a biomarker for minimal residual disease (MRD) detection in acute leukemia.

METHODSMethylation specific-PCR technique was used to detect Id4 gene methylation in samples with different percentages of leukemia cells from leukemia cell line and bone marrows from leukemia patients in complete remission (CR).

RESULTSId4 gene methylation could be detected in samples containing 1% or lower leukemia cells. Frequency of Id4 gene methylation in acute lymphoblastic leukemia (ALL) patients in CR was 64.3% being higher than that in acute myeloid leukemia (AML) patients in CR. In 14 ALL patients with Id4 gene methylation, 8 relapsed in 12 months, while only one relapsed in 9 patients without Id4 gene methylation. In 8 AML patients with Id4 gene methylation, 5 relapsed in 12 months, while two relapsed in 20 AML patients with Id4 gene methylation.

CONCLUSIONId4 gene promoter methylation is a candidate of biomarker for MRD detection in acute leukemias.

Acute Disease ; Adolescent ; Adult ; Aged ; Cell Line ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia ; diagnosis ; genetics ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; genetics ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; genetics ; Young Adult

To observe the influence of tumor necrosis factor-alpha (TNF-alpha) on differentiation of rat mesencephalic neural stem cells (NSCs), the numbers of neurons, astrocytes and oligodendrocytes generated from NSCs were analyzed after differentiation for 3 days by using immunocytochemistry technique. The results show that: (1) TNF-alpha enhanced the proportions of neurons and oligodendrocytes in progeny of NSCs; and (2) TNF-alpha induced the proliferation of oligodendrocytes derived from NSCs, but the proliferation of astrocytes was not influenced by TNF-alpha. We conclude that the TNF-alpha could influence the application of NSCs.
Animals ; Animals, Newborn ; Astrocytes ; cytology ; Cell Differentiation ; physiology ; Cell Proliferation ; Mesencephalon ; cytology ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Oligodendroglia ; cytology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; physiology