Aim To investigate the changes in the proliferation and apoptosis of Siha cells after knocking down Rho GTPase-activating protein 30(ARHGAP30).Methods After designing specific shARHGAP30 primers and connecting them to the pLKO.1 vector,we transformed them into Escherichia coli competent cells,then co-transfecting them with lentiviral helper plasmids into HEK-293T cells.We collected and filtered cell supernatant to obtain the vi-rus to infect Siha cells.RT-qPCR and Western blot were used to detect knockdown efficiency,as well as changes in the expression of Bax and Bcl-2 after trans-fection.The CCK-8 method was employed to measure the proliferation level of cells after knockdown.Results After successful construction of a lentiviral plasmid with knockdown of the ARHGAP30 gene and establish-ment of stably transfected Siha cells,ARHGAP30 tran-scription and translation(P＜0.01)in Siha cells de-creased,Bax/Bcl-2 significantly decreased(P＜0.01),indicating decreased apoptosis and increased cell proliferation(P＜0.01).Conclusions This study suggests the involvement of ARHGAP30 in the proliferation and apoptosis of Siha cells,and regulating the ARHGAP30 gene may interfere with the occurrence and development of cervical cancer.

Objective To evaluate the effect of SiO2-ZrO2 slurry coating on surface performance of zirconia ceramic.Methods Seventy pre-sintered zirconia discs were randomly divided into seven groups with 10 discs per group.Sample discs in each group received one of the following seven different surface treatments, namely, sintered(group AS), sand blasting after sintered(group SB), coated with slurry of mole ratio of SiO2 to ZrO2 2: 1(group 2SiO2-1ZrO2), coated with slurry of mole ratio of SiO2 to ZrO2 1: 1(group 1SiO2-1ZrO2), coated with slurry of mole ratio of SiO2 to ZrO2 1 : 2(group 1SiO2-2ZrO2), coated with slurry of mole ratio of SiO2 to ZrO2 1 : 3(group 1SiO2-3ZrO2), coated with slurry of mole ratio of SiO2 to ZrO2 1:4(group 1 SiO2-4ZrO2).Profilometer, X-ray diffractometer(XRD), energy dispersive spectrometer, scanning electron microscopy(SEM) were used to analyze surface performance.Results The surface roughness of the discs in group AS was lower than those in the other groups[(0.33 ± 0.03) μm] (P＜0.05), there was no statistically significant difference(P＞0.05) among group 2SiO2-1ZrO2[(3.85±0.38) μm], group 1SiO2-1ZrO2[(3.78±0.56) μm] and group 1SiO2-2ZrO2[(4.06 ± 0.48) μtm], and no difference(P＞0.05) was observed between group 1SiO2-3ZrO2 [(1.02±0.09) μm] and group 1SiO2-4ZrO2[(1.53±0.23) μm] either.However, surface roughness in all coating groups was higher than those in group SB[(0.86 ± 0.05) μm] (P＜0.05).According to the XRD pattern, group AS and all coating groups consisted of 100％ tetragonal airconia and monoclinic zirconia was detected at surface of group SB.Contents of surface silicon of coating groups increased significantly,however, no silicon was detected at sample surface of group AS and group SB.SEM showed that zirconia grains of coating exposed since part of silicon was etched by hydrofluoric acid, a three-dimensional network of intergrain nano-spaces was created.Conclusions SiO2-ZrO2 slurry coating could make surface of zirconia rough and increase Si content without creating monoclinic zirconia.

OBJECTIVETo investigate the effects of polybrominated diphenyl ether-153 (BDE-153) exposure during lactation period on the calcium ion (Ca(2+)) concentration and calcium-activated enzyme levels in cerebral cortical cells among adult rats and to provide a scientific basis for the study on the developmental neurotoxicity of BDE-153.

METHODSForty newborn male rats were randomly and equally divided into four groups according to their body weights and litters: 1, 5, and 10 mg/kg BDE-153 groups and olive oil solvent control group. On postnatal day 10 (PND 10), the BDE-153 groups were administrated BDE-153 (0.1 ml/10 g body weight) by intraperitoneal injection, while the olive oil solvent control group was given an equal volume of olive oil. Two months later, these rats were decapitated, and the cerebral cortex was separated quickly on an ice-cold dish. The Ca(2+) concentration in cerebral cortical cells was measured by flow cytometry. The activities of calcineurin (CaN) and Ca(2+)-Mg(2+)-ATP enzyme were determined by colorimetric method. The mRNA and protein expression of calpain-1 and calpain-2 was measured by real-time quantitative PCR and Western blot.

RESULTSThe mean fluorescence intensities of intracellular Ca(2+) in control group and 1, 5, and 10 mg/kg BDE-153 groups were 10.83, 1.48, 1.93, and 0.62, respectively; the 1, 5, and 10 mg/kg BDE-153 groups had significantly lower intercellular Ca(2+) concentrations than the control group (P < 0.05). The activities of CaN and Ca(2+)-Mg(2+)-ATP enzyme and mRNA and protein expression of calpain-1 showed no significant differences between the 1, 5, and 10 mg/kg BDE-153 groups and control group (P > 0.05). The protein expression of calpain-2 increased as the dose of BDE-153 rose. Compared with the control group (mRNA: 0.81±0.26; protein: 0.15±0.07), the 5 and 10 mg/kg BDE-153 groups had significantly higher mRNA expression of calpain-2 (5 mg/kg BDE-153 group: 1.16±0.52; 10 mg/kg BDE-153 group: 1.32±0.23) and significantly higher protein expression of calpain-2 (5 mg/kg BDE-153 group: 0.31±0.07; 10 mg/kg BDE-153 group: 0.37±0.06) (P < 0.05). The 10 mg/kg BDE-153 group had significantly higher protein expression of calpain-2 than the 1 mg/kg BDE-153 group (0.37±0.06 vs 0.22±0.07, P < 0.05).

CONCLUSIONCa(2+-) mediated calpain-2 activation may be one of the main mechanisms of BDE-153 neurotoxicity.

Animals ; Animals, Newborn ; Ca(2+) Mg(2+)-ATPase ; metabolism ; Calcineurin ; metabolism ; Calcium ; metabolism ; Calpain ; metabolism ; Cerebral Cortex ; metabolism ; Male ; Polybrominated Biphenyls ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the analgesic effect of sinomenine on the neuropathic pain rat model induced by SSNI, and discuss its impact on monoamine neurotransmitters in striatal extracellular fluid.

METHODMale SD rats were randomly divided into the sham operation group, the SSNI model group, the gabapentin group (100 mg x kg(-1)), the sinomenine high dose group (40 mg x kg(-1)) and the sinomenine low dose group (20 mg x kg(-1)). Mechanical hyperalgesia and cold pain sensitivity were evaluated by Von Frey hairs and cold spray. Striatum was sampled by microdialysis. High performance liquid chromatography-electrochemical detector (HPLC-ECD) were used to detect the content of such neurotransmitters as monoamine neurotransmitters noradrenaline (NE), dopamine (DA), 5-hydroxy tryptamine (5-HT) and their metabolites dihydroxyphenylacetic phenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA).

RESULTSSNI model rats showed significant improvement in mechanical withdrawal threshold and cold pain sensitivity, significant decrease in intracerebral NE and notable increase in DA, 5-HT and their metabolites. Compared with the model group, the sinomenine high dose group showed significant increase in mechanical withdrawal threshold at 60, 90, 180 and 240 min after abdominal administration (P < 0.01), significant decrease in cold pain sensitivity score during 30-240 min (P < 0.05). Sinomenine can significantly up-regulated NE content in striatal extracellular fluid during 45-135 min (P < 0.05), remarkably reduce DA content and DOPAC at 45, 75 and 135 min (P < 0.05), 5-HT content during 45-135 min, DOPAC during 75-165 min (P < 0.05), and 5-HIAA during 45-135 min (P < 0.05).

CONCLUSIONSinomenine has the intervention effect on neuropathic pain in SSNI model rats. Its mechanism may be related to disorder of monoamine neurotransmitters in striatal extracellular fluid.

Analgesics ; pharmacology ; Animals ; Biogenic Monoamines ; metabolism ; Disease Models, Animal ; Extracellular Fluid ; drug effects ; Male ; Morphinans ; pharmacology ; Neostriatum ; pathology ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; drug effects ; injuries ; metabolism ; pathology

OBJECTIVETo investigate the impact of sub-chronic Aluminium-maltolate [Al(mal)3] exposure on the catabolism of amyloid precursor protein (APP) in rats.

METHODSForty adult male Sprague-Dawley (SD) rats were randomly divided into five groups: the control group, the maltolate group (7.56 mg/kg BW), and the Al(mal)3 groups (0.27, 0.54, and 1.08 mg/kg BW, respectively). Control rats were administered with 0.9% normal saline through intraperitoneal (i.p.) injection. Maltolate and Al(mal)3 were administered to the rats also through i.p. injections. Administration was conducted daily for two months. Rat neural behavior was examined using open field tests (OFT). And the protein expressions and their mRNAs transcription related with APP catabolism were studied using enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR).

RESULTSThe expressions of APP, β-site APP cleaving enzyme 1 (BACE1) and presenilin-1 (PS1) proteins and their mRNAs transcription increased gradually with the increase of Al(mal)3 doses (P<0.05). The enzyme activity of BACE1 in the 0.54 and 1.08 mg/kg Al(mal)3 groups increased significantly (P<0.05). The expression of β-amyloid protein (Aβ) 1-40 gradually decreased while the protein expression of Aβ1-42 increased gradually with the increase of Al(mal)3 doses (P<0.05).

CONCLUSIONResult from our study suggested that one of the possible mechanisms that Al(mal)3 can cause neurotoxicity is that Al(mal)3 can increase the generation of Aβ1-42 by facilitating the expressions of APP, β-, and γ-secretase.

Amyloidogenic Proteins ; genetics ; metabolism ; Animals ; Drug Administration Schedule ; Environmental Pollutants ; administration & dosage ; toxicity ; Gene Expression Regulation ; drug effects ; Male ; Organometallic Compounds ; administration & dosage ; toxicity ; Pyrones ; administration & dosage ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the expression levels of cdk5, p35 and p53 genes in arsenic trioxide (As2O3O)-induced neuron apoptosis and to explore the potential mechanism.

METHODSThe cultured primary rats' neurons were divided into 5 groups, which were exposed to 0, 1, 5, 10 micromol/L As2O3 and dimethyl sulfoxide (DMSO) for 8 h, respectively. The cell viability and cell apoptosis were detected by MTT colouration methods and flow cytometry, respectively. The real-time fluorescence quantitative PCR was used to measured the expression levels of cdk5, p35 and p53 genes.

RESULTSThe cell viability inhibition rates were 16.77%, 19.72% and 27.81% in 1, 5, 10 micromol/L As203 groups, respectively. Compared to the untreated group and DMSO group, the cell apoptosis rates were significantly increased in 5 and 10 micromol/L As2O3 groups (P < 0.05). The expression levels of cdk5, p35 and p53 genes increased with the exposure doses of AsO3. However, there were no significant differences in p35 gene expression between different dose subgroups (P > 0.05). There were significant differences in cdk5 and p53 gene expression between different dose subgroups (P < 0.05). The expression levels of cdk5 gene in 5 and 10 micromol/L As2O3 groups were significantly higher than those in untreated group and DMSO group (P < 0.05). The expression levels of p53 gene in 1, 5 and 10 micromol/L As2O3 groups were significantly higher than that in untreated group (P < 0.05). The expression level of p53 gene in 10 mciromol/L As2O3 group was significantly higher than that in DMSO group (P < 0.05).

CONCLUSIONCdk5, p35 and p53 genes may involve in the process of As2O3-induced neural cell apoptosis.

Animals ; Apoptosis ; drug effects ; Arsenicals ; Cells, Cultured ; Cyclin-Dependent Kinase 5 ; genetics ; metabolism ; Neurons ; drug effects ; metabolism ; Oxides ; toxicity ; Phosphotransferases ; genetics ; metabolism ; Rats ; Tumor Suppressor Protein p53 ; genetics ; metabolism

BACKGROUNDCerebral alveolar echinococcosis (CAE) grows infiltratively like a malignant tumor, causing great harm to the human body. It is possible to display mass lesions of CAE using various imaging systems, but regarding the infiltrating proliferation active regions, it is difficult to evaluate its actual range using conventional magnetic resonance imaging (cMRI). This research focused on proton magnetic resonance spectroscopy ((1)HMRS) techniques to find the mass and infiltration zone of CAE. We explored the marginal zone (MZ) of CAE nearly close to the actual infiltrating scope, to provide reliable images for clinical purposes, to overcome shortcomings of cMRI, to formulate beneficial clinical surgical plans and assess prognosis.

METHODSBetween September 2005 and May 2011, 15 patients who were suffering from CAE (36 effective lesions altogether) were examined by (1)HMRS at the first affiliated hospital of Xinjiang Medical University. Multi-voxel (1)HMRS was acquired with a 1.5T MRI scanner. Concentrations and the ratios of the metabolites of CAE were calculated. Furthermore, changes in the concentrations of the metabolites containing N-acetyl-aspartic-acid (NAA), choline (Cho), creatine (Cr), lipids and lactate (Lip + Lac) and the ratios of Cho/Cr, NAA/Cr, (Lip + Lac) /Cr were compared in the substantial region, 0 - 10 mm MZ, and 11 - 20 mm MZ of the infiltration zone, as well as the corresponding contralateral part of the normal brain parenchyma area (control group).

RESULTSIn this study, the ratios of Cho/Cr in the substantial region, 0 - 10 mm MZ of infiltration zone and the control group were 1.78 ± 0.70, 1.90 ± 0.54, and 0.78 ± 0.15, respectively; the ratios of NAA/Cr were 1.60 ± 0.20, 1.80 ± 0.42, 2.24 ± 0.86, respectively; the ratios of (Lip + Lac)/Cr were 25.69 ± 13.84, 25.18 ± 16.03, and 0.61 ± 0.15, respectively. From the control group, 11 - 20 mm MZ to 0 - 10 mm MZ and the substantial region of CAE, the concentrations of the metabolites showed that NAA and Cho decreased gradually and markedly. But (Lip + Lac) increased gradually and markedly. The ratios of Cho/Cr and NAA/Cr, (Lip + Lac)/Cr were statistically significant (P < 0.0083) between the substantial region and the control group, as well as between the 0 - 10 mm MZ and the control group. The ratios of Cho/Cr and NAA/Cr, (Lip + Lac)/Cr displayed no statistically significant differences (P > 0.0083) between the substantial region and the 0 - 10 mm MZ.

CONCLUSIONSThere was a pathological spectrum surrounding the infiltration zone of CAE. Multi-voxel 1HMRS has great clinical value for discerning the main lesion and the infiltration zone of CAE.

Adult ; Central Nervous System Infections ; pathology ; Echinococcosis ; pathology ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Young Adult

ObjectiveTo investigation the clinical features and immunophenotypes of mantle cell lymphoma (MCL).MethodsThe clinical data of 23 patients of MCL were reviewed prospectively.Age,sex,Ann-Arbor staging,symptoms and bone marrow biopsies were analyzed.Serum lactat dehydrogenates (LDH) level,CD5,CD20 and CyclinD1 were determined.ResultsThe median age was 62 years old (range 44-74),15 patients(65.2％)were more than 60 years old.Among 23 patients,the male-to-female ratio was 4.4∶1,Twenty-one patients (91％) presented with advanced stage (Ann Arbor stage Ⅲ 12 cases and Ⅳ 9 cases) at initial diagnosis.Twenty patients (87.0 ％) were lymph node involvement.The serum LDH level was high in 11 patients.CD5 was positively expressed in 14 (60.8 ％) of all patients.CD20 was positively expressed in 21 (91.3 ％) of all patients.Cyclin D1 was over-expression in 19 (82.6 ％) of all patients.ConclusionThe MCL shows a predilection for occurrence in older males.The majority of patients are advanced stage disease (Ann Arbor stage Ⅲ/Ⅳ ) are at initial diagnosis. Most of patients display lymph node involvement manifestations.Bone marrow infiltration is frequent.The immunophenotype of MCL resembles the mature B-lymphocyte (CD+20),with coexprssion of the T-cell antigen CD5.It has been demonstrated that CyclinD1 are over express in this histotype of MCL.CyclinDl over-expression could be considered as hallmark of MCL.

OBJECTIVETo investigate the changes of mitochondria membrane potential and cytoplasma cytochrome C as the mechanism of neuron apoptosis induced by B(a)P.

METHODSPrimary neurons were dissociated from cerebral cortex of 1 - 3 days old SD rats and cultured with DMEM incubator at 37 degrees C. After 5 days' cultivation, the neurons were added S9 and B(a)P, and the concentrations of treated B(a)P were 0, 10, 20 and 40 micromol/L respectively. After administering of B(a)P, the neurons were cultivated for 40 hours. Apoptosis rate was measured by flow cytometry using Annexin V-FITC and propidium iodide (PI) staining, and the changes in mitochondrial potential (DeltaPsim) were tested with Rhodamine fluorescence (R2123) technique. Preparation of cytosolic extracts by centrifugation. Western blotting analysis was used to evaluate the level of cytochrome C of cytoplasm.

RESULTSThe apoptotic rate of neuron increased in both the middle dose group and the high dose group compared with controls, and had a dose-response tendency with the concentration of B(a)P. Moreover mitochondrial potential decreased in a dose dependent manner. There was a negative correlation between DeltaPsim and the apoptotic rate of neurons (r = -0.763, P < 0.05); Western blotting analysis showed cytoplasmic cytochrome C level increased significantly, which was positively related with neuron apoptosis (r = 0.831, P < 0.01).

CONCLUSIONLoss of mitochondria membrane potential and increase of cytoplasma cytochrome C may be the main cause of neuron apoptosis induced by B(a)P.

Animals ; Apoptosis ; drug effects ; Benzo(a)pyrene ; toxicity ; Cells, Cultured ; Cytochromes c ; metabolism ; Membrane Potential, Mitochondrial ; Mitochondria ; drug effects ; metabolism ; Neurons ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of Erzhi Pill (二至丸,EZP) on nerve cell apoptosis in senescence model rats.

METHODSThe rats model of senescence was established by peritoneal D-galactose injection combined with thymusectomy. Forty SD rats were randomized into four groups, the normal control group, the senescence model group, the EZP treated group, and the vitamins treated group, 10 in each group. The rats were made into senescence model except those in the normal group. In the same time of D-galactose injection, the rats were treated respectively with distilled water, EZP 4.32 g/kg, and vitamins E and C 0.06 g/kg daily for 6 weeks via intragastric infusion. The index of main viscera (as brain, testis, etc.), serum levels of superoxide dismutase (SOD) activity, and total anti-oxidation capacity (T-AOC) were measured after a 6-week treatment. Meanwhile, the cerebral cortex neuronal apoptosis proportion and mitochondrial membrane potential (MMP) were detected by flow cytometry.

RESULTSBoth EZP and vitamins E and C treatments showed effects on increasing testis index and serum level of T-AOC, reducing the percentage of neuronal apoptosis in the cerebral cortex, and elevating MMP in the aging rats model.

CONCLUSIONSEZP could inhibit the cerebral cortex neuron apoptosis and maintain the mitochondrial function in the senescent process of rats induced by peritoneal D-galactose injection combined with thymusectomy. It also shows antioxidation effect to some extents.

Aging ; blood ; drug effects ; Animals ; Antioxidants ; metabolism ; Apoptosis ; drug effects ; Cerebral Cortex ; cytology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Matrix Metalloproteinases ; metabolism ; Neurons ; cytology ; drug effects ; enzymology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood