Objective To investigate the effect of MT-1 and MT-2 in liver damage in chronic inorganic arsenate exposure mice and to explore the mechanism of arsenic-induced liver damage. Methods The male Kunming mice were randomly divided into control and exposed groups. The control group was given ordinary feed and tap water. The exposed group was given ordinary feed and 300 mg/L of sodium arsenite solution by drinking water. After 10 months of treatment, the activity of serum alanine aminotransferase (ALT), aspartate aminotransferase(AST) and globulin (Glb) content were determined. The total RNA was extracted by the TRIzol-phenol-chlorofor-method from the liver tissue. The quantity of the RNA was determined by spectrophotometry and its purity was judged at a ratio of A260/A280. Then real-time PCR(RT-PCR) was used to measure the mRNA expression of MT-1 and MT-2. Results Compared with the control group, serum ALT, AST activity and Glb content were higher and the MT-1 and MT-2 mRNA content was lower in the exposed group (P

The morphological indexes of the Scuophularia ningpoensis seedlings including the longth, diameter and weight were measured, clustering analysis was used to set up the standard quality grading of seedlings of S. Ningpoensis by SPSS. Field experiment was carried out to measure the indicators of plants growth and development, the yield and the quality. The results showed that the growth and yield of class I seedlings were better than those of class II and III. The content of main active ingredients was affected barely by seedlings classification. To ensure the quality, class II seedlings or above should be used for plantation. The established quality classification standard of S. ningpoensis seedlings was scientific and feasible, and provides the basis for the standardized cultivation of S. ningpoensis.
Drugs, Chinese Herbal ; analysis ; standards ; Quality Control ; Scrophularia ; chemistry ; classification ; growth & development ; Seedlings ; chemistry ; growth & development

BACKGROUND:Cytological studies show that bone marrow mesenchymal stem cel s play an important role in postmenopausal osteoporosis mechanism.
OBJECTIVE:To study the osteogenic differentiation in vitro of bone marrow mesenchymal stem cel s from ovariectomied osteoporotic rats.
METHODS:The osteoporotic animal model was established by performing ovariectomy in the 6-month-old female Sprague-Dawley rats. There were four groups:bone marrow mesenchymal stem cel s control group, bone marrow mesenchymal stem cel s osteoporosis group, bone marrow mesenchymal stem cel s osteogenic induction group and oseogenesis induction group. Bone marrow mesenchymal stem cel s were isolated from the rats of control group and oseogenesis induction group by means of the whole bone marrow adherence method and cultured to the 3rd generation. Then the bone marrow mesenchymal stem cel s were used in al the experiments. Cel morphology was observed under the inverted phase contrast microscope, cel cycle and proliferation index of bone marrow mesenchymal stem cel s were detected by flow cytometry. After osteogenic induction, the expression level of alkaline phosphatase was detected, and the fornation of calcium nodes of bone marrow mesenchymal stem cel s were marked by alizarin red staining.
RESULTS AND CONCLUSION:The cel s in the osteogenic induction group and oseogenesis induction group had the morphology of osteobalsts, and the change of morphology of the cel s in the oseogenesis induction group was relatively tardiness. The proliferation index in the control group was higher than that in the osteoporosis group (P<0.05);expression level of alkaline phosphatase in the osteogenic induction group was significantly higher than that in the oseogenesis induction group (P<0. 05), and the control group was significantly higher than the oseogenesis group (P<0.05). The alizarin red staining of the cel s in the osteogenic induction group was positive, while negative in the control group and the oseogenesis group;the staining in the osteogenic induction group was stronger than that in the oseogenesis induction group. These findings indicate that both the proliferative potential and the osteogenic potential of bone marrow mesenchymal stem cel s from the ovariectomized osteoporotic rats are decreased, which may be related with the ostoeporosis pathogensis of ovariectomied rats.

OBJECTIVETo investigate the effect of Antrodia cinnamomea on gene expression related to aortal endothelial injury of rats with hyperlipidemia.

METHODFifty SD rats were randomly divided into five groups: the normal control group (NG), the model group (MG), the antrodia cinnamomea groups of low, middle and high doses (AC-LG, AC-MG, AC-HG, 250, 500, 1 000 mg x kg(-1)). The rats were fed with high-fat diets to establish the hyperlipidemia model. After the drug administration for 10 weeks, their serum lipid, SOD, MDA and ox-LDL, LOX-1, P38 MAPK and NF-kappaB mRNA and protein expression were respectively determined, and the aortal endothelial injury was observed under electron microscope.

RESULTIn the model group, the contents of TC, TG and LDL-C significant increased (P < 0.01), whereas the content of HDL-C significant decreased (P < 0.01). Compared with the model group, both the AC-M group and the AC-H group showed reduction in endothelial injury and significant decrease in the content of TC, TG and LDL-C (P < 0.05 or P < 0.01). The content of HDL-C increased, but with no significant difference. SOD activity in serum remarkably increased (P < 0.05 or P < 0.01), MDA and ox-LDL levels dramatically decreased (P < 0.05 or P < 0.01).

CONCLUSIONA. cinnamomea can alleviate endothelial lipid injury by inhibiting the expressions of LOX-1, P38MAPK and NF-kappaB in aorta and better protect aortal endothelial cells from oxidative lipid injury.

Animals ; Antrodia ; chemistry ; Aorta ; drug effects ; metabolism ; ultrastructure ; Atherosclerosis ; blood ; genetics ; prevention & control ; Biological Products ; pharmacology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Endothelium, Vascular ; drug effects ; metabolism ; pathology ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; drug effects ; Hyperlipidemias ; blood ; genetics ; prevention & control ; Lipoproteins, LDL ; blood ; Male ; Malondialdehyde ; blood ; Microscopy, Electron ; NF-kappa B ; blood ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Scavenger Receptors, Class E ; blood ; genetics ; metabolism ; Superoxide Dismutase ; blood ; Triglycerides ; blood ; p38 Mitogen-Activated Protein Kinases ; blood ; genetics ; metabolism

Aim To investigate the role of phospho-lipase C epsilon 1 ( PLCE1 ) in modulating the apoptot-ic mechanism in esophageal cancer ( Eca ) cells. Methods Eca cell lines, OE33 and CP-C cells were cultured to assess the expression of PLCE1 . siRNA suppress expression of PLCE1 . The expression of PLCE1 and p53 was evaluated by quantitative real time PCR and Western blot. Methylation analyses of p53 were performed by bisulfite conversion of genomic DNA. Apoptosis was assessed by flow cytometry. Results OE33 and CP-C cells expressed high levels of PLCE1 . Knockdown of PLCE1 markedly increased the expression and hypomethylation of p53 , and in-creased the frequency of apoptosis. Conclusion PLCE1 suppresses apoptosis of Eca cells via promoting p53 promoter methylation and inhibiting expression of p53 .

Objective To establish the high myopic choroidal neovascularization (CNV) in guinea pigs and to investigate the role of (matrix metalloproteinase-2,MMP-2) and tissue inhibitor-2 of metalloproteinase (TIMP-2) in high myopic CNV.Methods Seventy-two 2-week-old guinea pigs were randomized into control group (n =36) and high myopia group (n =36).Right eyes were indued form deprivation high myopia for 6 weeks.Thirty guinea pigs were randomly selected in each group,and CNV were induced in the right eyes br the 532 nm laser.MMP-2 and TIMP-2 expressions were investigated by immunohistochemistry pre-laser and 7,14,21,28,35 days after laser induction,respectively,while MMP-2 and TIMP-2 relative expression levels in retinal pigment epithelium (RPE)-choroid-sclera complex were detected by real-time PCR.Integral opitical density (IODs) of positive expression and mRNA relative expression levels of these factors were performed by statistical analyses.Results The expressions of MMP-2 and TIMP-2 were up-regulated in the two groups after laser photocoagulation through immunohistochemistry and real-time PCR examinations.Expression of MMP-2 peaked at 21 d and TIMP-2 at 28 d,respectively.IODs of positive expression and mRNA relative expression levels of MMP-2 were higher in high myopia group than those in control group at each inspective time point,and the differences were statistically significant (P ＜ 0.05).TIMP-2 expression was significantly reduced in high myopia group compared with control group before laser photocoagulation (P＜0,05),while there was no significant difference between the two groups at each time point after laser photocoagulation.Conclusions MMP-2 and TIMP-2 were closely related to the formation of high-myopic CNV.Balance disorders of MMP-2 and TIMP-2 might participate in the occurrence and development of high-myopic CNV.

Aim To investigate the activation of prote-ase-activated receptor 2 ( PAR2 ) in regulation of the expression of epidermal growth factor receptor ( EGFR) and apoptosis of lung cancer ( LC) cells. Methods LC cells A549 and its EGFR-silenced counterpart were incubated in the medium added with tryptase. Quanti-tative RT-PCR and Western blotting were used to de-tect the expression of EGFR in A 5 4 9 cells . The apop-tosis and Bax/Bcl-xL of cells were also recorded. Re-sults Treating A549 cells with tryptase in the culture for 48 hrs resulted in a marked increase in the expres-sion of EGFR in A549 cells. Marked decreases in a tryptase dose-dependent manner in apoptotic A549 cells were detected in the presence of tryptase. Expo-sure to tryptase markedly decreased the levels of Bax and increased the levels of Bcl-xL in A549 cells, which were not shown in EGFR-deficient A549 cells. Conclusion Tryptase can increase the expression of EGFR in LC cell line, A549 cells, which protects A549 cells from apoptosis, increases Bcl-xL, and sup-presses Bax in A549 cells.

Objective To investigate the regulative effect of cyclooxygenase-2 (COX-2) 3'-UTR in HT29 colon cancer cells. Methods Total RNA was extracted from HT29 colon cancer cells and used as a template to amplify COX-2 gene by reverse transcription polymerase chain reaction. Using PCR technique to construct a series of luciferase reporter gene expression vectors containing various regions of the 3'-UTR of COX-2 (including the whole gene region, gene region between 1 to 156 bp, gene region between 1 to 347 bp, gene region between 1 to 1006 bp, gene region between 156 to 347 bp and gene region between 157 to 2217 bp). These reporter gene expression vectors and pRL-SV40 were co-transfected in HT29 colon cancer cells by Lipofectamine 2000. The relative luciferase activity of the HT29 colon cancer cells was tested. All data were analyzed via t test. Results The luciferase activity was reduced by 70.4%, 37.4%, 64.8% and 24.2% in HT29 colon cancer cells transfected with the whole COX-2 gene region, gene region between 1 to 156 bp, gene region between 1 to 347 bp and gene region between 156 to 347 bp, respectively (t = 6.13, 7.73, 9.75, 3.92, P < 0.05). No obvious changes of luciferase activity were observed in HT29 colon cancer cells transfected with gene regions between 1 to 1006 bp and between 157 to 2217 bp (t = 0.13, 0.01, P > 0.05). Conclusions A region between 156-347 bp in the 3'-UTR of COX-2 has been found which can down-regulate the expression of COX-2 with the cooperation of the ARE element. The 3'-UTR of COX-2 contains several control elements that regulate the expression of COX-2 in colon cancer cells.

Adult ; Coal Mining ; Disability Evaluation ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; physiopathology ; Respiratory Function Tests

Adult ; Aged ; Cartilage Diseases ; diagnosis ; therapy ; Humans ; Laryngeal Cartilages ; Laryngeal Diseases ; diagnosis ; therapy ; Male ; Middle Aged