OBJECTIVETo study the flavanoids extracted from onion on the blood-brain barrier (BBB) permeation, and their effects on primary cultured neuron cell proliferation and apoptosis of SD rats using ethanol reflux method.

METHODSThe brain microvascular endothelial cells (BMVECs) were first successfully primary cultured. Then rats BMVECs and astrocytes (ACs) were co-cultured to establish the in vitro BBB model. The flavanoids were extracted from onion using ethanol reflux method. The model was verified by transmission electron microscopy (TEM) and trans-epithelial electric resistance (TEER). The flavanoids permeability was tested using high performance liquid chromatography (HPLC). Meanwhile, rat neuron cells were cultured and exposed to H2O2 and flavanoids. Their effects on the cell proliferation and apoptosis were observed using MTT assay. The injury of neuron DNA was analyzed using single-cell gel electrophoresis (SCGE) and immunofluorescent assay.

RESULTSThe in vitro BBB model was successfully established by TEM and TEER. Results of HPLC proved flavanoids extracts could effectively permeate the BBB with the permeability of 60.58%. The extractive at 10 - 20 microg/mL showed obvious inhibition on the apoptosis of neuron cells induced by H2O2, and attenuated the injury of neuron DNA.

CONCLUSIONSThe flavanoids extracted from onion ethanol reflux method could effectively penetrate the BBB. They also showed obvious inhibition on the H2O2 induced neuron cell apoptosis and DNA injury.

Animals ; Animals, Newborn ; Apoptosis ; Astrocytes ; cytology ; drug effects ; Blood-Brain Barrier ; drug effects ; Cells, Cultured ; DNA Damage ; Endothelial Cells ; cytology ; drug effects ; Flavonoids ; pharmacology ; Hydrogen Peroxide ; Neurons ; cytology ; drug effects ; Neuroprotective Agents ; pharmacology ; Onions ; chemistry ; Rats

Tilianin was separated and authenticated from the seeds of Dracocephalum moldavia, a Uygur medicine, by chromatographic technique and spectroscopic method. The purity of tilianin is more than 98% determined by HPLC area normalization method. Thin layer chromatography (TLC) method was used to separate tilianin from D. moldavia by mixture of chloroform-methanol (5: 1) as a developing solvent on high performance silicagel precoated plate (SGF254) and using aluminium trichloride as a chromogenic agent for qualitative identification of D. moldavia. To establish a HPLC method for quantitative analysis of D. moldavia, tilianin was used as a Quantitative marker and separated on a C18 (4.6 mm x 250 mm, 5 μm) column with acetonitrile-01% formic acid (25: 75) as the mobile phase and detected at 330 nm. The calibration curve of tilianin displayed ideal linearity over the range of 0.617 2-123.44 μg x mL(-1) with a regression equation of Y = 33.773X - 0.824 8 (r = 1). The average recovery of tilianin was 101.0% with RSD of 3.7%. The RSD values of intra-day and inter-day precision were less than 2%. The content of tilianin in 4 batches of the authenticated semen of D. Moldavia was between 0.016 and 0.187 mg x g(-1). The qualitative and quantitative method established is suitable for the quality evaluation and assessment of semen of D. Moldavia.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Flavonoids ; chemistry ; isolation & purification ; standards ; Glycosides ; chemistry ; isolation & purification ; standards ; Lamiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Quality Control

Objective To investigated the related factors which affecting the severity degrees of post-traumatic stress disorder(PTSD) in trailsferred casualty after Wenchuan Earthquake.Methods Taking PTSD symptoms self-assessment scale(PCL-C) to involve 386 wounded who sufiered 40 days after the earthquake disaster,from 11 hospitals and were transferred to Chongqing city.Multi-stage cluster sampling method was used.354 valid questionnaires were recovered to explore the relevant factors affecting the severity on the symptoms of PTSD.Results This survey contains 354 subjects,with male 154(43.6%),female 200(56.4%),age 43.76±21.22,nation alities:Han people 236(66.7%),Qiang people 114(32.2%),others4(1.1%),and marriage status as unmarried 92(26.1%),married 253(71.7%),others 9(2.2%).The wounded women PTSD have more seriotis symptoms than men,and there were differences between them in repeated and disturbing dreams of this stressful experience(t=2.46,P=0.014),a strong sense of psychological suffering annoyance (t=2.02,P=0.044),having difficulty concentrating(t=2.04,P=0.042),being"super-alert"(t=2.465,P=0.014) etc,also in the total scores(t=2.489.P=0.013)(P<0.05).The PTSD symptoms of wounded degree in who had been buried in Earthquake were more serious than those not been buried.There were significant difierences between them in the iterns as:avoid thinking about or talking about a stressful experience(t=2.661,P=0.008),avoid activities or situations that could fecall the stressful experience(t=2.705,P=0.007),trouble remembering important parts of a stressful experience (t=2.775,P=0.006),feeling emotionally numb or being unable to have loving feelings for those close to you(t=3.017,P=0.003),feeling as if your future will somehow be cut short(t=2.979,P=0.003) and total scores(t=3.175,P=0.002).The wounded that witnessed someone be buried or died in earthquake,in PTSD symptoms,were more serious than those without.In the items of feeling distant or cut off from other people,feeling emotionally numb or being unable to have loving feelings for those close to you,feeling irritable or having angry outbursts, there were significant difference(P<0.05-0.01).There were significant difrerences among the average score of the three core symptoms(F=3.350,P<0.001),among the three core symptoms as well as between the three core symptoms and total score respectively.Conclusion There were differences in the severity degrees of PTST in transferred casualty under difrerent exposed conditions.Correlations were also found between each of the symptoms.

Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA, Viral ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effects of pSilencer 1.0-U6-siRNA-STAT3 on the growth of PC3 and LNCaP cells.

METHODSThree pairs of DNA template coding siRNA were synthesized against STAT3 to reconstruct pSilencer 1.0-U6-STAT3-siRNA, which was transfected into PC3 and LNCaP cells. The STAT3 expression in PC3 cells and LNCaP were transfected with pSilencer 1.0-U6-siRNA-STAT3, and it was detected by Western blot and Northern blot. MTT and FCM were used to observe the growth-inhibiting ratio and apoptosis in PC3 cells.

RESULTSWestern blot and Northern blot analyses demonstrated that pSilencer 1.0-U6-siRNA-STAT3 could significantly inhibit the expression of STAT3 in PC3 and LNCaP cells; MIT and FCM results showed that it could suppress the growth of PC3 cells and LNCaP and induce apoptosis of PC3 cells in vitro.

CONCLUSIONPSilencer 1.0-U6-siRNA-STAT3 could significantly inhibit STAT3 expression, suppress the growth of PC3 and LNCaP cells and induce the apoptosis of PC3 cells.

Apoptosis ; Cell Line, Tumor ; Humans ; Male ; Plasmids ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; RNA, Small Interfering ; STAT3 Transcription Factor ; biosynthesis ; genetics ; Transcription, Genetic

Objective To investigate the prevalence of self-reported suicidal behavior in university students and to compare the related risk factors between two selected groups. Methods A questionnaire survey on situation of suicidal behavior and risk factors was conducted among 9808 university students. Results The overall rate of suicide attempts among university students was 1.7%. Factors as suicidal attitude, having associates who had suicidal experience and negative life events, were significantly different between groups with or without suicide attempts. Risk factors on suicidal behavior were as follows: high frequency of feeling hopeless in the last year (OR=4.23,95% CI: 3.05-5.85) , first degree relatives with suicide behavior (OR=2.85, 95% CI: 1.72-4.73),having associates with suicide behavior (OR=2.82,95%CI: 2.05-3.88), having had sexual experience (OR=2.73,95%CI: 1.88-3.96), daily live being influenced by the existing psychological problems in the last month (OR=2.53, 95% CI: 1.80-3.56) , being female (OR=2.47, 95% CI: 1.77-3.45) and having other relatives with suicide behaviors (OR=2.36, 95% CI: 1.63-3.41). Conclusion Psychological, biological and social factors had influenced on suicidal behaviors among university students which all called for effective intervention to be implemented.

OBJECTIVETo study the effect of cyclooxygenase -2 selective inhibitor celecoxib on the expression of major vault protein ( MVP) in the brain of rats with status epilepticus and its possible roles in the treatment of refractory epilepsy.

METHODSSixty adult male Sprague-Dawley rats were randomly assigned to blank control (n=16), epilepsy model (n=22) and celecoxib treatment groups (n=22). After the status epilepticus was induced in rats by injecting lithium and pilocarpine, each group had 16 rats enrolled as subjects. Immunohistochemical method and Western blot method were used to detect the expression of MVP in the frontal cortex and hippocampus.

RESULTSThe expression of MVP was significantly higher in the epilepsy model group than in the control group (P<0.01). The expression of MVP in the celecoxib treatment group was significantly decreased compared with the epilepsy model group, but it was still higher than in the control group (P<0.01).

CONCLUSIONSCelecoxib could decrease the expression of MVP in brain tissue of rats with status epilepticus, suggesting that it is promising for the treatment of intractable epilepsy.

Animals ; Blotting, Western ; Brain ; metabolism ; Celecoxib ; pharmacology ; therapeutic use ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; drug therapy ; metabolism ; Vault Ribonucleoprotein Particles ; analysis

Objective To investigate the effects of Bushen Huoxue acupuncture method on the behavioral changes and the proliferation of endogenous neural stem cells (NSCs) in hippocampus of Alzheimer disease (AD) model mice. Methods 18 male SAMP8 mice, seven months old, were randomly divided into acupuncture group, non-acupoint control group, and model control group. And another age-matched 6 male SAMR1 mice were prepared as normal control group. Mice in acupuncture group were intervened by acupuncture method in the acupoints of "Shenshu", "Baihui", "Xuehai", and "Geshu". Mice in non-acupoint control group were treated by stimulating the fixed non-point under the bilateral rib, while mice in model control group and normal control group were raised without special treatment but administered the stimulation of catching with the same time and the same stimulus intensity. All treatrment lasted for 8 weeks. After the intervention, the learning and memory abilitities and brain hippocampus Brd U positive cells of all groups were detected. Results Compared with the nomal control group, mice in the model control group had longer escape latency and less time spent in former platform quadrant (P＜0.05); the number of Brd U positive cells decreased significantly (P＜0.05). Compared with the model control group and the non-acupoint control group, mice in the acupuncture group had shorter escape latency and more time spent in former platform quadrant (P＜0.05); the number of Brd U positive cells increased significantly (P＜0.05). Conclusion Bushen Huoxue acupuncture method can improve the learning and memory abilities of SAMP8 mice AD model by inducing the proliferation of hippocampal endogenous NSCs.

The aim of the present study was to investigate the effect of ERK on 17beta-estradiol (E(2)) inhibition of vascular smooth muscle cell (VSMC) proliferation in rats after vascular injury. Common carotid artery balloon-injury (Inj) model was established in ovariectomized rats (OVX). Female SD rats were randomly divided into 4 groups: OVX, E(2)+OVX, OVX+Inj, and E(2)+OVX+Inj groups. The thickness of the vessels, the plasma content of NO, and the expression of ERK, phosphorylated ERK as well as eNOS protein were measured. The results showed that compared with OVX, the vessel wall was significantly thickened and the plasma content of NO was significantly decreased in OVX+Inj group. E(2) significantly decreased the vessel thickness but increased the plasma NO content after balloon injury. E(2) inhibited the expression of ERK, phosphorylated ERK and induced the eNOS expression. There is a positive correlation between plasma NO content and eNOS protein expression, while there is a negative correlation between plasma NO content and the thickness of vessel. The plasma NO content and the expression of ERK protein were negatively correlated. These results suggest that E(2) increases the vascular eNOS protein expression and NO release, leading to the inhibition of VSMC proliferation after balloon injury by inhibiting the ERK and phosphorylated ERK protein expression.
Animals ; Carotid Artery, Common ; pathology ; Catheterization ; adverse effects ; Cell Proliferation ; drug effects ; Estradiol ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; physiology ; Female ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; physiology ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; metabolism ; Ovariectomy ; Phosphorylation ; Rats

The defect or block of apoptosis is an important factor involved in the drug resistance of tumor cells. Blm gene plays a great role in DNA damage and repair. This study was aimed to explore the relationship of blm gene expression with cell cycle and apoptosis after Jurkat DNA damage. The apoptosis rate and change of cell cycle were detected by flow cytometry, the expression level of blm mRNA in Jurkat cells was determined by semi-quantitative RT-PCR. The results indicated that after induction with 0.4 g/L of mitomycin C (MMC) for 24 hours the apoptosis rate of Jurkat cells were (11.42±0.013)%, and (66.08±1.60)% Jurkat cells were arrested in G2/M phase. After induction for 48 hours, the apoptosis rate of Jurkat cells declined from (11.42±0.013)% to (8.08±0.27)%, and cell count of Jurkat cells arrested in G2/M phase decreased from (66.08±1.60)% to (33.96±1.05)%. When induced with 0.4 g/L of MMC for 24 hours, the apoptosis rate of fibroblasts and the percentage of fibroblasts in G2/M, G0-G1 and S phase all showed no significant change until 48 hours. The range of apoptosis rate and the change of cell percentage in three phases were significantly different between Jurkat cells and fibroblasts (p<0.01). Expression level of blm mRNA in Jurkat cells was remarkably higher than that in normal fibroblasts (p<0.01), at 48 hours expression level of blm mRNA was remarkably higher than that at 24 hours. The 2 groups showed clear difference of blm mRNA expression after treated by MMC (p<0.01). It is concluded that the blm gene may play a significant role in repair of DNA damage of Jurkat cells after MMC induction. Abnormal expression of blm is correlated to the drug resistance of leukemia cells.
Apoptosis ; Cell Cycle ; DNA Damage ; drug effects ; DNA Repair ; drug effects ; Humans ; Jurkat Cells ; Mitomycin ; pharmacology ; RecQ Helicases ; genetics