Objective To observe the correlation between nuclear factor ?B(NF-?B) expression and apoptosis after hypoxia-ischemia brain damage(HIBD).Methods Forty-eight 7-day newborn Sprague-Dawley(SD) rats were randomly divided into 2 groups: control group (n=24) and HI group (n=24).The expression of NF-?B in the hippocampus was detected by immunohistochemical techniques.The apoptosis of the hippocampal cells was detected by terminal deoxynucleotidyltransferase-mediated 2-deoxyuridine 5′-triphosphate-biotin nick end labeling(TUNEL)straining.Results The expression of NF-?B in hippocampal cells increased at 6 h after HIBD,peaked at 48 h,and lasted till 72 h when compared with control group(Pa

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

AIM: To explore whether strontium ranelate ( Sr ) promotes osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) through the Hedgehog/Gli1 signaling pathway.METHODS:BMSCs were isolated from 4-week-old rats by adherent culture and induced to differentiate into osteoblasts .According to the experimental purposes , the cells were exposed to different concentrations of Sr , cyclopamine ( Cy, an inhibitor of Hedgehog receptor ) or Gli1-siR-NA.The expression of Gli1 and Runx2 in the cells was detected by Western blotting .The activity of alkaline phosphatase ( ALP) was measured by the method of colorimetry , and the mineralized nodules were observed under microscope with aliz-arin red staining .RESULTS:Exposure to Sr at concentrations of 0.1 to 5 mmol/L for 7 d markedly increased the expres-sion of Gli1 in the BMSCs , and the increase in Gli1 expression was the most obvious following Sr exposure at concentration of 3 mmol/L.Cy at concentration of 10 μmol/L inhibited Sr-induced up-regulation of Gli1 expression.Transfection of the BMSCs with Gli1-siRNA not only obviously inhibited Sr-induced up-regulation of Gli1 and Runx2 ( a downstream protein of Gli1) expression, but also antagonized Sr-induced enhancement of ALP activity and the formation of mineralized nodules . CONCLUSION:The Hedgehog/Gli1 pathway is involved in Sr-induced osteogenic differentiation of rat BMSCs .

Objective To study the expressions of vascular endothelial growth factor (VEGF) and Ki-67 antigen and their correlation in human brain astrocytorna. Methods Immunohistochemistry with SP method was employed to detect the expressions of VEGF and Ki-67 in 60 human brain astrocytoma tissue (grade Ⅰ-Ⅱ 28 cases, grade Ⅲ 20 cases, grade Ⅳ 12 cases) and 30 normal tissues adjacent to the tumors. Results VEGF and Ki-67 were not expressed in the normal brain tissues adjacent to the astrocytoma, but their expressions were detected in all the astroeytoma tissues of different grades (P<0.05). High-grade astrocytomas showed significantly higher VEGF and Ki-67 expression than low-grade astrocytomas (P<0.05), and a positive correlation was noted between VEGF and Ki-67 expressions (r=0.310, P<0.05). Conclusion VEGF and Ki-67 expressions can provide important evidence for evaluating the malignancy and clinicopathologieal grading of human astrocytoma.

OBJECTIVETo study whether effect of aspirin plus low-dose diethylstilbestrol is more effective and safer than high diethylstilbestrol dose alone on prevention of ovariectomy-induced osteopenia and dyslipidemia.

METHODSThirty-eight 4-month-old female SD rats were divided into baseline (BAS) group (n=6), sham operation group (n=8) and ovariectomy (OVX) group (n=24). The OVX group was further divided into vehicle treatment group (n=8), diethylstilbestrol (30 μg/kg•d) treatment group (OVX+D30 group, n=8), and aspirin (9 mg/kg•d) plus diethylstilbestrol (10 μg/kg•d) treatment group (OVX+A-D10 group, n=8). Their left tibiae were collected for the bone histomorphometric analysis in undecalcified sections. Left femurs were collected for the bone mineral density measurement.

RESULTSThe body weight and serum cholesterol were increased, while uterine weight and cancellous bone mass were decreased in OVX rats compared with the SHAM group. Cancellous bone mass was significantly increased, while body weight and bone resorption parameters were decreased in both A-D10 and D30 treatment group compared with OVX group. The rats treated with A-D10 showed significantly increased in bone formation parameters and decreased in serum triglyceride compared with the D30-treated rats.

CONCLUSIONAspirin plus low-dose diethylstilbestrol can effectively prevent osteopenia by reducing bone resorption, and is thus a better treatment modality for preventing dyslipidemia than high-dose diethylstilbestrol alone.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; therapeutic use ; Aspirin ; pharmacology ; therapeutic use ; Biomarkers ; blood ; Body Weight ; drug effects ; Bone Density ; Bone Diseases, Metabolic ; blood ; prevention & control ; Bone and Bones ; drug effects ; Diethylstilbestrol ; pharmacology ; therapeutic use ; Drug Evaluation, Preclinical ; Drug Therapy, Combination ; Dyslipidemias ; blood ; prevention & control ; Estrogens, Non-Steroidal ; pharmacology ; therapeutic use ; Female ; Organ Size ; drug effects ; Ovariectomy ; Rats ; Uterus ; drug effects

OBJECTIVETo study the protective mechanisms of the astragaloside against ischemia-reperfusion lung injury in rats.

METHODSIschemia-reperfusion lung injury was induced in SD rats. Astragalus armour glucoside was dissolved in 1% of sodium carboxymethyl cellulose at different concentrations (8, 6, and 3 mg/ml) was intragastrically administered in the rats at the dose of 1 ml/100 g. Cellular and subcellular structural changes in the lung tissue were observed at the end of the experiment using optical and transmission electron microscope, with the wet/dry ratio of the lung tissue and myeloperoxidase (MPO) activity measured.

RESULTSThe wet/dry ratio and myeloperoxidase activity in the lung tissue were significantly higher in the model group than in the sham-operated group (P<0.05), and were significantly lowered by the treatment with astragalus armour glucoside at different doses (P<0.01 or 0.05), and the effect was especially obvious in rats receiving a moderate dose. Pulmonary capillary expansion, erythrocyte leakage and exudate in the alveolar space with obvious pathological changes in the type I and II epithelial cells were observed in model group. Pulmonary capillary expansion was reduced in rats treated with high, medium and low dose of Astragalus armour glucoside, and the medium dose group showed the most obvious effect, in which no edema fluid in the alveolar space or erythrocyte leakage was found with also reduced type II lung epithelial cell degranulation.

CONCLUSIONAstragaloside has obvious antioxidant effect in rats with ischemia-reperfusion lung injury, and a medium dose produces the best effect.

Animals ; Female ; Ischemic Preconditioning ; Lung ; drug effects ; pathology ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; pathology ; Saponins ; pharmacology ; therapeutic use ; Triterpenes ; pharmacology ; therapeutic use

OBJECTIVETo study the expression of decoy receptor 3 (DcR3) and its relationship with apoptosis and prognosis in hepatocellular carcinoma (HCC).

METHODSThe expression of DcR3 protein in 43 cases of HCC and 16 cases of non-cancerous liver (including cirrhotic liver tissue and normal liver tissue adjacent to cavernous hemangioma) was studied by immunohistochemistry (using EnVision method). The status of apoptosis was evaluated by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) technique. Statistic analysis was carried out to assess the correlation between DcR3 expression, apoptotic index (AI) and clinicopathologic parameters.

RESULTSDcR3 protein was expressed in the cytoplasm of HCC cells. The positivity rate of DcR3 in HCC was 74.42% (32/43), which was significantly higher than that in the non-cancerous group (43.75%, P < 0.05). The positivity rate of DcR3 in HCC with metastasis detected within 20 months of diagnosis was 100% (22/22). This was significantly higher than that in HCC without metastasis (52.94%, P < 0.01). The DcR3 expression in HCC also correlated with serum alpha-fetoprotein level (r = 0.444, P < 0.01) and presence of tumor embolus in portal vein (r = 0.414, P < 0.01). However it had no relationship with the patient's age, sex, cirrhotic status, liver capsule invasion, number of tumor nodules and histologic differentiation (P > 0.05). The AI in HCC (0.78 +/- 0.64)% was significantly lower than that in the non-cancerous group [(3.32 +/- 1.81)%, P < 0.01]. The AI in clinical TNM stage I and II tumors (1.03 +/- 0.69)% was significantly higher than that in stage III and IV tumors [(0.52 +/- 0.48)%, P < 0.01]. The AI in HCC without metastasis (1.10 +/- 0.72)% was significantly higher than that in HCC without metastasis [(0.44 +/- 0.27)%, P < 0.05]. The AI correlated with serum alpha-fetoprotein level (r = -0.468, P < 0.01), presence of tumor embolus in portal vein (r = -0.434, P < 0.01) and liver capsule invasion (r = -0.331, P < 0.05). On the other hand, it had no relationship with patient's age, sex, cirrhotic status, number of tumor nodules and histologic differentiation (P > 0.05). The AI in DcR3-positive group (including both HCC and non-cancerous tissues) was significantly lower than that in DcR3-negative group (P < 0.01).

CONCLUSIONSThe expression of DcR3 in HCC correlates with apoptosis of tumor cells and may play a crucial role in tumor pathogenesis and progression. DcR3 protein expression and AI may also serve as important biologic indicators in predicting prognosis of HCC.

Adult ; Aged ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism ; physiology ; Retrospective Studies ; Young Adult

OBJECTIVETo explore the mechanism of transcription regulation of the liver-selective genes responsible for cell communication.

METHODSTissue-selective Affymetrix probe sets (3919 probes in total) were clustered by functional categories. Liver-selective cell communication (LSCC) genes were selected for further analysis. The 500-bp upstream sequences of all the LSCC genes were extracted for predicting the transcription factor binding sites (TFBS) of known transcription factors (TFs) using 3 programs; literature mining was then performed for these LSCC genes and TFs, and the transcription regulatory network were constructed.

RESULTSThe binding sites of 50 and 72 transcription factors were predicted from the upstream sequences of 23 LSCC genes by two programs respectively. Among them, 18 transcription factors were found in common. The top 10 TFBS sequences were basically consistent to the predicted TFs. Literature mining indicated that LSCC genes and TFs were closely related to such terms as albumin, diabetes, glucose, lipid, metabolism, and JNK, in addition to those associated with hepatic tissue and TFs. These observations suggested that LSCC genes and TFs were involved in the regulation of glucose and lipid metabolism, binding and transport, coagulation signal cascades, inflammatory response, etc. PPP2R1B, which was out of the network, showed a partial functional similarity to DUSP10 in the network.

CONCLUSIONSLSCC genes and the predicted TFs may be involved in the regulation of many important functions of the liver, which are integrated into a sophisticated transcription regulatory network. JUN may be the key target for regulation, and PPP2R1B is presumed to participate in the regulation of JUN.

Binding Sites ; genetics ; Cell Communication ; genetics ; Gene Expression Profiling ; Gene Expression Regulation ; Gene Regulatory Networks ; genetics ; Humans ; Liver ; cytology ; metabolism ; Models, Biological ; Transcription Factors ; genetics ; Transcription, Genetic

OBJECTIVEThe objective of this study is to investigate the influence of zirconia content which is 0-30.0% weight percentage of matrix on translucency of zirconia-toughened alumina glass-infiltrated ceramics.

METHODSSeven groups were divided according to different weight percentage of zirconia (0, 2.5%, 5.0%, 7.5%, 10.0%, 20.0% and 30.0%). After sintering, infiltrating and polishing, spectral transmittance was determined with spectrophotometer under D65 standard source. Contrast ratio was also tested by whiteness colorimeter.

RESULTSWith mass fraction of zirconia increasing from 0 to 30.0%, spectral transmittance reduced from 0.406% to 0.058%, while contrast ratio value increased from 0.849 +/- 0.005 to 1.015 +/- 0.006. When zirconia content was 10.0%, contrast ratio was 0.990 +/- 0.008. When it was more than 10.0%, transmission rate of the downward trend and contrast ratio of the rising trend became flat.

CONCLUSIONZirconia content has a direct impact on translucency of zirconia-toughened alumina glass-infiltrated ceramic, which is essentially opaque when zirconia content is 10.0%. When mass fraction of zirconia is more than 10.0%, the influence of zirconia content is reduced.

Aluminum Oxide ; Ceramics ; Dental Materials ; Dental Porcelain ; Glass ; Zirconium

OBJECTIVEAge, body weight and dose have been shown as important influencing factors for sodium valproate plasma concentrations. However it is unclear whether there is interaction among them and whether the interaction could influence sodium valproate plasma concentrations. This study aimed to evaluate the influence of age, body weight and dose on plasma concentrations of sodium valproate and the interaction among them.

METHODSOne hundred and thirty-two children with epilepsy (age: 4 months-6 years, weight: 5-25 kg) were enrolled. Sodium valproate was administered at the dosage of 10-30 mg/kg/d. Plasma concentrations of sodium valproate were measured by high-performance liquid chromatography 3-5 days after administration. The relationship of sodium valproate plasma concentrations with age, body weight, and dose of sodium valproate was examined using variance analysis, pearson correlation analysis and stepwise regression analysis.

RESULTSAge (F=8.630, P<0.01), body weight (F=3.650, P<0.05) and dose of sodium valproate (F=11.720, P<0.01) were influencing factors for sodium valproate plasma concentrations. The interaction between age and oral dose (F=2.484, P<0.05) and the interaction of age and body weight with oral dose (F=4.923, P<0.01) had significant effects on sodium valproate plasma concentrations. Stepwise regression analysis showed that dose of sodium valproate and body weight were entered to the regression equation.

CONCLUSIONSAge, body weight and dose of sodium valproate as well as the interactions between age and dose and between age, body weight and dose were influencing factors for valproate plasma concentrations.

Age Factors ; Anticonvulsants ; blood ; Body Weight ; Child ; Child, Preschool ; Epilepsy ; drug therapy ; Female ; Humans ; Infant ; Male ; Regression Analysis ; Valproic Acid ; administration & dosage ; adverse effects ; blood