BACKGROUNDOur previous study had cloned two glioma cell lines SWOZ1 and SWOZ2 isolated from parental glioma cell line SWO38. The 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) resistance of SWOZ1 was higher than that of SWOZ2. Since O6-methylguanine-DNA methyltransferase (MGMT) was thought to be closely related to BCNU resistance in glioma, this study aimed to explore the function of MGMT in glioma resistant to BCNU.

METHODSA BCNU resistant glioma cell line SWOZ2-BCNU was established. The expression of MGMT was detected in SWOZ1, SWOZ2 and SWOZ2-BCNU. Small interferencing RNA targeting MGMT was used to silence the expression of MGMT in resistant cell lines SWOZ1 and SWOZ2-BCNU. The cytotoxicity of BCNU to these cells was measured using the cell counting kit-8 assay. Statistical analysis was carried out by one-way analysis of variance in statistical package SPSS 13.0.

RESULTSThe resistance of SWOZ1 and SWOZ2-BCNU against BCNU was 4.9-fold and 5.3-fold higher than that of SWOZ2. The results of quantitative RT-PCR and Western blotting confirmed that MGMT was both significantly increased in SWOZ1 and SWOZ2-BCNU compared to SOWZ2. After transfection with small interferencing RNA targeting MGMT, a decreased level of MGMT mRNA expression in SWOZ1 and SWOZ2-BCNU for more than 75% compared to negative control was found and confirmed by Western blotting. As a result, the resistance against BCNU was reversed for about 50% both in the BCNU-resistant cell lines SWOZ1 and SWOZ2-BCNU.

CONCLUSIONSSilencing MGMT with specific small interferencing RNA can reverse the BCNU resistant phenotype in these glioma cell lines. MGMT may play an important role both in intrinsic and acquired BCNU-resistance in glioma.

Blotting, Western ; Carmustine ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Glioma ; genetics ; metabolism ; Humans ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; RNA, Small Interfering ; Reverse Transcriptase Polymerase Chain Reaction ; Sincalide ; metabolism

OBJECTIVE: By summarize the characteristics of death cases caused by road traffic accident, to provide information and data for prevention of traffic accident. METHODS: To retrospectively analyze 4148 death cases caused by road traffic accident in Shenzhen. The characteristics studied include the age and sex of the dead, the cause of death, time and place of the accidents and vehicle types, etc. RESULTS: The death was mainly male and the proportion of male to female is 2.45:1; the accidents mainly occured in 6:00-8:00 and 18:00-2:00; 72% of the accidents took place on the main suburban roads. The proportions for each traffic modes of the death were: 44% of walking, 19% of bike, 15% of motorbike. Most traffic accidents were induced by truck, 83.2% of the death causations were severe head injury and 13.3% were complex injuries. CONCLUSION The death cases of road-traffic accident in Shenzhen has obvious characteristic and maybe is preventible.
Accidents, Traffic/prevention & control* ; Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Cause of Death ; Child ; Child, Preschool ; China/epidemiology* ; Craniocerebral Trauma/epidemiology* ; Female ; Humans ; Infant ; Male ; Middle Aged ; Retrospective Studies ; Sex Factors ; Young Adult

Objective To observe the improving effect in joint function of moderate skeletal fluorosis treated by traditional Chinese medication(main ingredient was Strychnine).MethodsFrom December 2007 to July 2009,120 moderate skeletal fluorosis patients met the inclusion criteria were divided into the treatment group(60 cases)and the control group(60 cases)in the skeletal fluorosis hospital of Xinzhou,the treatment group was given basic treatment and traditional Chinese medication,the control group wa8 given basic treatment and placebo.The treatment lasted 12 weeks,follow up 24 weeks.Before treatment,after treatment 4 weeks,8 weeks,12 weeks,36 weeks,a third party evaluate comprehensive function of both upper and lower limb and joint dysfunction.Results The main effect of both drugs was statistically significance in the scores of the upper forearm in the finger by touching the posterior contralateral ear, upper arm touched by the finger back in the opposite corner subscapularis function, lower limb function and single-joint dysfunction(F values were 4.08,14.32,35.81,13.02, all P<0.05), the main effect of time also was significant (F values were 82.63,72.82,277.33,328.16, all P<0.05),①the upper forearm in the finger by touching the posterior contralateral ear functions:At the time of 8,12 weeks,scores of the treatment group were lower than those of before treatment and control group (all P<0.05);At the time of 36 weeks,scores of the treatment group were lower than that 12 weeks(all P<0.05);At the time of 8,12,36 weeks, scores of the control group were lower than those of before treatment(all P < 0.05);②upper arm function, namely fingers touching the opposite corner subscapularis:At the time of 4,8,12 weeks, scores of the treatment group were lower than those of before treatment(all P<0.05); At the time of 36 weeks, scores of the treatment group were lower than that 12 weeks(all P<0.05); At the time of 8,12,36 weeks, scores of the treatment group were lower than those of the control group (all P<0.05);③Lower extremity functions: At the time of 8,12 weeks,scores of the treatment group were lower than those of before treatment and control group (all P<0.05);At the time of 36 weeks, scores of the treatment group were lower than that 12 weeks(all P<0.05) ; At the time of 8,12,36 weeks, scores of the control group were lower than those of before treatment (all P<0.05);④single joint functions:At the time of 4,8,12 weeks,scores of the treatment group were lower than those of before treatment(all P<0.05); At the time of 36 weeks,scores of the treatment group were lower than that 12 weeks(all P<0.05) ; At the time of 8,12,36 weeks, scores of the control group were lower than those of before treatment(all P<0.05);At the time of 4,8,12,36 weeks, scores of the treatment group were lower than those of control group(all P<0.05);⑤At the end of treatment and follow-up,the improvement rate in joint functions in the treatment group were 88.33% (53/60),93.33% (56/60); the control group were 28.07%(16/57),40.35%(23/57), (Fisher test, P<0.01,X2=56.21, P<0.01). ConclusionTraditional Chinese medication(its main ingredient is Strychnine), an effective drug for improving joint dysfunction in patients suffering from moderate skeletal fluorosis, is simple and effective.

Objective To explore the pharmacokinetics of single and multiple oral 20 mg escitalopram in healthy Chinese volunteers. Methods A total of 12 subjects participated in the study. Escitalopram 20 mg was given orally once on day 1 and days 8 to 14 in the fast condition. Sequen-tial blood samples were collected over 144 hours on day 1 and 14 and a predose sample was obtained on day 12 to 14. Escitalopram concentrations in plasma were determined by a validated HPLC fluorescence method. The pharmacokinetic parameters were calculated with DAS software. Results Escitalopram disposition on oral administration is characterized by a two-compartment pharmacokinetic model. The mean t1/2 is 41.09 h, Cavis (76.4±26.8) μg·L-1, AUCss is (1832.4±642.4) μg·h ·L-1,AUC0-t, and AUC0-∞are (4765.9±2171.0) and (5385.6±2851.2) μg ·h·L-1, respectively; tmaxis (3.2±1.3) h;t1/2 is (41.1±17.7) h;CLis 5.0 L·h1. The mean accumulation index of AUC ( RAUC) is ( 1.2 ±0.3 ). Conclusion Escitalopram pharmacokinetics in healthy Chinese subjects given 20 mg once daily dosing regimen were characterized by a two-compartment pharmacokinetic model. The state -concentration oc-curre after 7 days of continuously dosing. There is no accumulation after continuously dosing.

BACKGROUNDTo explore the effect of human osteopontin (hOPN) on the proliferation, transmigration and expression of matrix metallproteinase-2 (MMP-2) and matrix metallproteinase-9 (MMP-9) in osteosarcoma (OS) cells in vitro.

METHODSThe prokaryotic-expression vector of hOPN was produced. hOPN was then subcloned into E. coli BL21 (DE3) cells and purified with ProBond trade mark Columns. The proliferation, cell cycle and the expression of cyclin A in OS cells were investigated by using MTT assay, flow cytometry and Western blot respectively. The transmigration of OS cells was checked by using transwell cell culture chamber. The micro-pore-filter-membrane system was used to study the chemiotaxis of hOPN to OS cells. The levels of total protein were examined according to Coomassie Brilliant Blue manuals. The expression of MMP-2 and MMP-9 were evaluated by detecting the volume of degradation of gelatin on SDS-PAGE gel.

RESULTSThe prokaryotic-expression vector of hOPN and purified hOPN protein were achieved hOPN promoted OS cells proliferation in a dose-dependent manner, and stimulated cyclin A expression in OS cells to accelerate cell division cycle. hOPN facilitated the trans-membrane migration of OS cells. hOPN also enhanced the secretion of MMP-2 and MMP-9 in OS cells.

CONCLUSIONhOPN could stimulate cyclin A expression in OS cells. hOPN has chemiotaxis to OS cells and increases their transmigration. hOPN enhances the secretion of MMP-2 and MMP-9 in OS cells.

Bone Neoplasms ; pathology ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; secretion ; Matrix Metalloproteinase 9 ; secretion ; Osteopontin ; Osteosarcoma ; pathology ; Sialoglycoproteins ; pharmacology

OBJECTIVETo explore the effect of muscovite on the quality of gastric ulcer healing.

METHODGastric ulcers were produced in male rats by serosal application of acetic acid. Rats were gavaged for 14 days with saline, omeprazole and muscovite starting 3 days after ulcer induction. Then the tissue and blood samples were obtained and measured.

RESULTIn the muscovite group, restored mucosa thickness increased, cystically dilated glands decreased, microvessels in connective tissue increased, the secretion of mucus, hexosamine, PGE2, EGF, bFGF were enhanced, and the express of EGFR was stronger.

CONCLUSIONMuscovite can promote the gastric ulcer healing and improve the quality of gastric ulcer healing.

Aluminum Silicates ; pharmacology ; Animals ; Dinoprostone ; blood ; Fibroblast Growth Factor 2 ; metabolism ; Gastric Mucosa ; pathology ; physiopathology ; Hexosamines ; metabolism ; Male ; Materia Medica ; pharmacology ; Mucus ; secretion ; Rats ; Rats, Wistar ; Receptor, Epidermal Growth Factor ; metabolism ; Stomach Ulcer ; chemically induced ; pathology ; physiopathology

OBJECTIVETo evaluate the effect of Valeriana officinalis var latifolia(VOL) on expression of transforming growth factor beta 1 (TGF-beta 1) in hypercholesterolemic rats and study its possible mechanisms.

METHODDietary-induced hypercholesterolemia was induced in male Wistar rats by given 4% cholesterol and 1% cholic acid diet for 16 weeks. Changes of serum lipid, urinary albumin, renal function and Mesangial matrix index were assessed. Moreover, immunohistochemical stain for TGF-beta 1 and type IV collagen were performed.

RESULTVOL could reduce the serum levels of total cholesterol, low density lipoprotein, urinary albumin and serum creatinine. Light microscopy and immunohistochemical stain revealed that in the same time of lowing serum lipid, Mesangial matrix index was significantly reduced, accompanied by decreased expression of TGF-beta 1 and type IV collagen.

CONCLUSIONVOL has the protective effect on lipid-induced nephropathy, and the inhibition of TGF-beta 1 expression might be the mechanism of VOL on renal protection.

Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Hypercholesterolemia ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; Male ; Phytotherapy ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; Valerian ; chemistry

OBJECTIVETo observe the changes of auditory steady-state response (ASSR) threshold in guinea pigs with posterior semicircular canal fistula.

METHODSIn 10 guinea pigs, a window was opened in the posterior semicircular canal of the left ear with the right ear serving as the control for testing the ASSR under anesthesia with pentobarbital sodium.

RESULTSThe mean and standard deviation of ASSR threshold (dB SPL) at 0.5, 1, 2, and 4 kHz in the left ear was 35.00+/-14.33, 25.50+/-12.37, 20.00+/-9.37 and 20.00+/-9.18, respectively, and was 31.00+/-16.19, 25.50+/-12.34, 18.00+/-6.96 and 18.50+/-6.71 in the right ear, respectively. Paired-sample t test showed no significant difference in the ASSR at the same frequency between the two ears.

CONCLUSIONSmall fistula by surgery causes no significant hearing loss in guinea pigs.

Animals ; Audiometry, Pure-Tone ; methods ; Auditory Threshold ; physiology ; Cochlea ; pathology ; Evoked Potentials, Auditory ; Fistula ; pathology ; physiopathology ; Guinea Pigs ; Labyrinth Diseases ; pathology ; physiopathology ; Semicircular Canals

OBJECTIVETo detect the cell-surface-expressed nucleolin and investigate its tumor suppressing effect on the growth of hepatocellular carcinoma cells.

METHODSTo detect cell-surface-expressed nucleolin in the hepatocellular carcinoma cells by immunofluorescence and flow cytometry. To down-regulate the nucleolin expression level in hepatocellular carcinoma cells by RNA interference. The tumor-suppressing effect of cell-surface nucleolin on hepatocellular carcinoma cells was assessed by MTT and transwell chamber assays.

RESULTSNucleolin was expressed in the nuclei, cytoplasm and on the cell surface of hepatocellular carcinoma cells. ShRNA markedly decreased the nucleolin expression level in the cytoplasm and on the cell surface (P < 0.01), but the nuclear nucleolin remained unchanged. After downregulation of cell-surface nucleolin, MTT assays showed that the cell growth rate of hepatocellular carcinoma cells in the shRNA interference group was significantly inhibited as compared with that in the control group (P < 0.01). The transwell chamber assay showed that the mean transmembrane cell number in the shRNA interference group was significantly lower than that in the control group.

CONCLUSIONThe results of this study show that downregulation of cell-surface nucleolin expression inhibits the growth of hepatocellular carcinoma cells in vitro.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cell Movement ; Cell Nucleus ; metabolism ; Cell Proliferation ; Cytoplasm ; metabolism ; Down-Regulation ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Phosphoproteins ; metabolism ; RNA Interference ; RNA, Small Interfering ; pharmacology ; RNA-Binding Proteins ; metabolism

To solve the issues of costly planting of facility cultivation method and inferior efficacy than wild herbs of Dendrobium officinale, the cliff epiphytic cultivation method was studied. To research the growth, agronomic traits, yield, polysaccharide and alcohol-soluble extract contents were measured on the D. officinale from different water regulation and cliff slope gradients treatments. The results showed that D. officinale epiphytic at 85 degrees-90 degrees cliff and sprayed water 1-2 h x d(-1) at the growing season can get better growth and obtain high yield, and the morphology has no different from wild cliff D. officinale, even in the environments without shade. The contents of polysaccharide and alcohol-soluble extract are closely related to the physiological ages, but significantly higher than the facility cultivation. It is possible that environmental stresses benefit the accumulation of polysaccharides, alcohol-soluble extract and other efficient ingredients.
Agriculture ; methods ; Dendrobium ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Polysaccharides ; analysis ; Water ; analysis