UNLABELLEDOBJECTIVE To study the in vitro effect and mechanism of Ginkgo biloba Extract 50 (GBE50) for inhibiting beta-amyloid (Abeta)-induced oxidative stress in rats' hippocampal neurons.

METHODSThe primary hippocampal neurons were cultured in vitro and divided into 4 groups, i. e. the normal control group (Ctrl), the Abeta group, the propanediol control group (PDO), and the six GBE50 concentrations groups (5, 10, 25, 50, 100, and 200 microg/mL). Excepted the Ctrl group, neurons were induced to oxidative stress by 20 gmolLAbeta25-35. The MTT and fluorescent probes labeling were used to observe the effect of GBE50 with different concentrations on the cell viability and the generation of intracellular reactive oxygen species (ROS) in neurons. Furthermore, Western blot was used to detect the cytoplasmic/total cytochrome C (Cyto C) ratio and total intracytoplasmal Cyto C, and the effect of the expression of oxidative stress-related protein Cyto C and activated Caspase-3 in three GBE50 concentrations groups (25, 50, and 100 microg/mL).

RESULTSCompared with the Ctrl group, the cell vitality was obviously lowered and intracellular ROS generation significantly increased after induction of 20 micromol/L Abeta25-35 (both P < 0.05). Compared with the Abeta group, the cell vitality was evidently improved after treated with different GBE50 doses. Except for 10 microg/mL, the cell vitality could be obviously elevated along with increased drug concentrations (P < 0.05). Meanwhile, the intracellular ROS generation decreased significantly in each GBE50 dose groups (P < 0.05). Abeta could increase the cytoplasmic/total Cyto C ratio and enhance the activated Caspase-3 expression significantly (P < 0.05). Compared with the Abeta group, among the three concentrations of GBE50, the Cyto C ratio was obviously lowered in the 100 microg/mL GBE50 group (P < 0.05), and the expression of activated Caspase-3 significantly decreased in 50 microg/mL and 100 microg/mL GBE50 groups (P < 0.05).

CONCLUSIONS20 micromol/L Abeta25-35 could induce the generation of intracellular ROS in hippocampal neurons. GBE50 could inhibit Abeta induced intracellular oxidative stress of neurons through lowering the cytoplasmic/total Cyto C ratio and inhibiting the activation of apoptosis protein Caspase-3 expression.

Amyloid beta-Peptides ; toxicity ; Animals ; Cells, Cultured ; Cytochromes c ; metabolism ; Hippocampus ; metabolism ; Neurons ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Peptide Fragments ; toxicity ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule.
Amino Acid Substitution ; Animals ; Mutation ; Sodium Channel Blockers ; Sodium Channels ; drug effects ; physiology ; Spider Venoms ; chemical synthesis ; genetics ; Structure-Activity Relationship ; Tetrodotoxin ; pharmacology

OBJECTIVETo explore the relationships between quality of life, negative life events, social support and suicide ideation among undergraduates in colleges.

METHODS3517 undergraduates in colleges were recruited by multistage stratified random clustered sampling method. Factors associated with suicide ideation were analyzed with logistic regression by scores of Beck Scale for Suicide Ideation(BSSI), Generic Quality of Life Inventory (GQOLI), Adolescent Self-rate Life Events Checklist (ASLEC), Social Support Rating Scale (SSRS) and a questionnaire on background information.

RESULTSThe rate of suicide ideation within 7 days was 14.1%, especially in females (15.96%), with single parent (23.79%) and disabled undergraduates (25.00%). The primary risk factors for suicide ideation were with low psychological function, material life, family/social support, lower availability of support and more negative life events.

CONCLUSIONThe prevalence of suicide ideation among these undergraduates was high, appropriate measures focusing on these risk factors should be implemented.

China ; epidemiology ; Cluster Analysis ; Female ; Humans ; Logistic Models ; Male ; Risk Factors ; Self-Injurious Behavior ; epidemiology ; Students ; psychology ; Suicide ; psychology ; Surveys and Questionnaires

BACKGROUNDNerve growth factor (NGF) is well-known for its important role in the development and maintenance of the nervous system. Along with its neurotrophic role, NGF has been detected in the testis of mouse, rat and human, suggesting an additional non-neurotrophic effect in the male reproductive system. The expression of β-NGF in the undescended testes (cryptorchidism) has not been detected at present. The aim of this study was to evaluate the expression of β-nerve growth factor mRNA and protein in experimental cryptorchidism.

METHODSA unilateral mechanical cryptorchidism model in the Sprague-Dawley rat was established and the expression of β-NGF with histologic changes in experimental cryptorchidism were investigated using one step quantitative real-time reverse transcription-polymerase chain reaction, in situ hybridization histochemistry, immunofluorescence and hematoxylin-eosin staining.

RESULTSThe expression of β-NGF mRNA and protein were both significantly decreased in the development of unmarred testis and cryptorchidism-induced testis, and the decrease of β-NGF in cryptorchidism-induced testis was far greater than that in uninjured testis.

CONCLUSIONFrom this investigation, we confirmed a lower expression of β-NGF in undescended testes than in the development of testis.

Animals ; Cryptorchidism ; genetics ; metabolism ; Male ; Nerve Growth Factor ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate risks and clinical effects of operative treatment for cervical vertebral fracture and dislocation associated with unilateral vertebral artery injury.

METHODSThis group consisted of 76 cases of closed cervical spine trauma combined with unilateral vertebral artery injury (23 cases of bilateral facet dislocation, 28 unilateral facet dislocation and 25 fracture). All patients underwent prospective examination of cervical spine MRI and vertebral artery two-dimensional time-of-flight (2D TOF) magnetic resonance angiography (MRA), and anterior cervical decompression. The healthy vertebral artery paths were evaluated before the surgery, and were protected during the surgery according to the anatomical signs.

RESULTSThere were no acute or chronic clinical damage symptoms in 76 cases after surgery. No neural damage symptoms were observed in patients with normal neural functions. The neural functions of incomplete paralyzed patients were improved in different grades.

CONCLUSIONSReliable anterior operation can produce good results for cervical fracture and dislocation with unilateral vertebral artery injury. Detecting the course of uninjured vertebral artery before operation and locating the anatomical site during operation are effective to avoid damaging vertebral artery of uninjured side.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cervical Vertebrae ; injuries ; Child ; Female ; Humans ; Joint Dislocations ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Vertebral Artery ; injuries ; Wounds, Nonpenetrating ; surgery

The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
Cell Separation ; methods ; Culture Media ; Pinellia ; physiology ; Protoplasts ; physiology ; Regeneration

OBJECTIVETo evaluate the benzene exposure level and cytopenia among the benzene exposed workers in Shanghai, China and to analyze the influential factors for the health of benzene-exposed workers.

METHODSA total of 3314 benzene-exposed workers, who were from 85 benzene-related enterprises selected by stratified random sampling based on enterprise sizes and industries, were included in the study. The time-weighted average (TWA) concentration of benzene in each workshop was measured by individual sampling and fixed point sampling, and the benzene exposure level in workshop was evaluated accordingly. The occupational health examination results and health status of benzene-exposed workers were collected.

RESULTSThe median of TW A concentrations of benzene was 0.3 mg/m3. The TWA concentrations measured at 7 ( 1.4%) of the 504 sampling points were above the safety limit. Of the 7 points, 3 were from large enterprises, 2 from medium enterprises, and 2 from small enterprises; 3 were from shipbuilding industry, 1 from chemical industry, and 3 from light industry. Of the 3314 benzene-exposed workers, 451 ( 13.6%) had cytopenia, including 339 males ( 339/2548, 13.3%) and 112 females ( 112/766, 14.6% ). There were significant differences in the incidence rates of leukopenia and neutropenia among the benzene-exposed workers of different sexes and ages (P<0.05); there were significant differences in the incidence rate of cytopenia among the benzene-exposed workers of different ages and working years ( P<0.05 ); there were significant differences in the incidence of neutropenia among the benzene exposed workers of different working years ( P<0.05).

CONCLUSIONMonitoring and intervention measures should be enhanced to protect the benzene-exposed workers in the large enterprises in shipbuilding industry and medium and private enterprises in chemical industry from occupational hazards.

Adolescent ; Adult ; Benzene ; toxicity ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Pancytopenia ; chemically induced ; epidemiology ; Young Adult

Animals ; Cells, Cultured ; Hepatic Stellate Cells ; metabolism ; Male ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the protective effect of six Kaixin San formulas on simulated cells model of depression, Alzheimer's disease and Parkinson's disease.

METHODThe in vitro simulated cells model of depression, Alzheimer's disease and Parkinson's disease was established by injuring SH-SY5Y cells with corticosterone (0.4 mmol x L(-1)) , injuring PC12 cells with neurotoxic amyloid peptide (Abeta25-35) (20 micromol x L(-1)) and injuring SH-SY5Y cells with 1-methyl-4-phenylpyridinium ion (MPP+) (250 micromol x L(-1)). The cell survival rate was assayed with MTT method and the degree of cell injury was detected with LDH.

RESULT100, 500 mg x L(-1) Dingzhixiao Wan prepared as mentioned in Beiji Qianjin Yaofang could significantly increase the survival ratio of SH-SY5Y cells injured by corticosterone and reduce LDH concentration released. All of the Kaixin San formulas could significantly increase the survival ratio of PC12 cells injured by Abeta25-35 and reduce LDH concentration released. Particularly, Kaixin San (10, 100, 500 mg L(-1)) prepared as mentioned in Beiji Qianjin Yaofang shown the best effect. And 500 mg x L(-1) Fushen Wan prepared as mentioned in Gujin Luyan could significantly increase survival ratio of SH-SY5Y cell injured by MPP and reduce LDH concentration released.

CONCLUSIONDingzhixiao Wan prepared as mentioned in Beiji Qianjin Yaofang could protect corticosterone-induced SH-SY5Ycells injury, showing a potential antidepressant effect. All of the six Kaixin San formulas could protect Abeta25-35-induced PC12 cells injury, but Kaixin San prepared as mentioned in Beiji Qianjin Yaofang shown the best potential effect for Alzheimer's disease. Fushen Wan prepared as mentioned in Gujin Luyan could protect MPP(+)-induced SH-SY5Y cells injury to some extent.

1-Methyl-4-phenylpyridinium ; toxicity ; Alzheimer Disease ; drug therapy ; Amyloid beta-Peptides ; toxicity ; Animals ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Neurons ; cytology ; drug effects ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Parkinson Disease ; drug therapy ; Peptide Fragments ; toxicity ; Rats

Zuotai (gTso thal) is a typical representative of Tibetan medicines containing heavy metals, but there is still lack of modem safety evaluation data so far. In this study, acute toxicity test, sub-acute toxicity test, one-time administration mercury distribution experiment, long-term mercury accumulative toxicity experiment and preliminary study on clinical safety of Compound Dangzuo were conducted in the hope of obtain the medicinal safety data of Zuotai. In the acute toxicity test, half of KM mice given the lethal dose of Zuotai were not died or poisoned, and LD50 was not found. The maximum tolerated dose of Zuotai was 80 g x kg(-1). In the subacute toxicity test, Zuotai could reduce ALT, AST, Crea levels in serums under low dose (13.34 mg x kg(-1) x d(-1)) and medium dose (53.36 mg x kg(-1) x d(-1)), with significant difference under low dose, and increase the levels of ALT, AST, MDA, Crea in serums under high dose (2 000 mg x kg(-1) x d(-1)); besides, the levels of BUN and GSH in serums reduced with the increase in dose of Zuotai, indicating a significant dose-effect relationship. In the one-time administration distribution experiment, the content of mercury in rat kidney, liver and lung increased after the one-time administration with Zuotai, with a significant dose-dependent relationship in kidney. In the long-term mercury accumulative toxicity experiment, KM mice were administered with equivalent doses of Zuotai for 4.5 months and then stopped drug administration for 1.5 months. Since the 2.5th month, they showed significant mercury accumulation in kidney, which gradually reduced after drug withdrawal, without significant change in mercury content in liver, spleen and brain and ALT, AST, TBIL, BUN and Crea in serum. At the 4.5th month after drug administration, KM mice showed slight structural changes in kidney, liver and spleen tissues, and gradually recovered to normal after drug withdrawal. Besides, no significant difference in weight gain was found between the Zuotai group and the control group. According to the findings of the clinical safety study of Dangzuo, after subjects administered Dangzuo under clinical dose for one month, their serum biochemical indicators, blood routine indicators and urine routine indicators showed no significant adverse change. This study proved that traditional Tibetan medicine Zuotai was slightly toxic, with a better safety in clinical combined administration and no adverse effects on bodies under the clinical dose and clinical medication cycle. However, long-term high-dose administration of Zuotai may have a certain effect on kidney.
Adult ; Animals ; Clinical Trials as Topic ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; toxicity ; Female ; Humans ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Medicine, Tibetan Traditional ; Mice ; Middle Aged ; Rats ; Rats, Wistar ; Young Adult