Objective To investigate the effects of dexamethasone(DEX)on the secretion of interleukin (IL)-17 and interferon(IFN)-γ and the proportion of Th17,Tc17,Th1 ,Tc1 cells in peripheral blood mononuclear cells(PBMCs)of patients with systemic lupus erythematosus(SLE). Methods Thirty hospitalized SLE patients were recruited and twenty-two healthy volunteers were recruited as healthy controls. PBMCs were separated from SLE patients and healthy controls and then was cultured in vitro by medium or PMA/Ionomycin or PMA/Ionomycin +dexamethasone for six hours. Four- color immunofluorescent staining and flow cytometric assay were used to analyze the percentage of Th17,Tc17,Th1,Tc1 cells in PBMCs. Concentrations of IL-17 and IFN-γ in plasma and the supernatants of PBMCs which were cultured for 24 hours were measured by enzyme linked immunosorbent assay (ELISA). Results The plasma concentrations of IL-17 and IFN-γwere elevated in SLE patients as compared to the controls(P ＜ 0.05). No significant differences were observed between patients and controls for the spontaneous production of IL-17 and IFN-γ or percentage of T subsets expressed by PBMCs. After the stimulation of PMA,compared with the controls,the level of IL-17 was significantly elevated in the supematants of PBMCs and the percentages of Th17 and Tc1 in SLE patients increased significantly(P ＜ 0. 05). However,there showed no significant differences between SLE patients and the controls for the percentages of Th1 and Tc17 cells. DEX could significantly decrease the production of IL-17(P ＜ 0. 01)and the percentages of Th17,Tc1 cells by the active PBMCs(P ＜ 0. 05). Conclusions There is abnormal expression of T subset cells and their cytokines in vivo of SLE patients. DEX can interfere with immunological pathological process in the cytokine network imbalance of SLE patients and shows powerful inhibition of IL - 17. Our results may provide some laboratory evidence for the clinical application of corticosteroids.

BACKGROUND: Osteogenic growth polypeptide (OGP) had clear effect on promoting osteoblast proliferation, differentiation and mature. OBJECTIVE: To explore the expression of OGP gene, which was transfected into rabbit bone marrow mesenchymal stem cells (BMSCs) and to evaluate the effects of OGP on differentiation of rabbit BMSCs. METHODS: pcDNA3.1-OGP was constructed using gene cloning and recombination techniques. Rabbit BMSCs were transfected with pcDNA3.1-OGP mediated by lipofectamine 2000. The transfection positive cell clones were selected with G418. The expression of OGP gene was detected using reverse transcription-polymerase chain reaction analysis on an mRNA level. Differentiation of pcDNA3.1-OGP transfected BMSCs into osteoblast lineage was observed. RESULTS AND CONCLUSION: The pcDNA3.1-OGP plasmid was constructed successful and OGP expression was detected in rabbit BMSCs. Hydroxyproline content was increased, and alkaline phosphatase activity was also increased. These indicate that pcDNA3.1-OGP transfected BMSCs expressed OGP, and could differentiate into osteoblast lineage.

Dental Enamel Hypoplasia ; classification ; etiology ; genetics ; Humans ; Molecular Biology

Aged ; Female ; Giant Cell Arteritis ; complications ; Humans ; Otitis Media with Effusion ; complications

Glycosyltransferases (GTs) catalyze the transfer of a sugar residue of an activated sugar donor to an acceptor molecule. Many families 1 GTs utilize an uridine diphosphate (UDP) activated sugar as donor in the glycosylation reaction, and most of these belong to a group of GTs referred to as the UGTs. The relationship between the degree of amino acid sequence identity and substrate specificity of the plant UGTs is highly complicated, and the prediction of substrate specificity based on phylogenetic analyses need to be improved by more biochemical characterization. This review summarizes the three dimensional structures of plant UGTs published in the Protein Data Bank (PDB), including the detailed substrate interactions with the sugar and receptor binding pockets and mutational analyses of some critical amino acids. It will be helpful for biochemical characterization the substrate specificity of the individual UGT, and lay the foundation for the enzymatic and genetic manipulation of plant UGTs in the future.
Amino Acid Sequence ; Glycosylation ; Glycosyltransferases ; chemistry ; Phylogeny ; Plant Proteins ; chemistry ; Plants ; enzymology ; Protein Structure, Tertiary ; Substrate Specificity ; Uridine Diphosphate ; chemistry

Objective To observe the apoptosis or oncosis of pancreatic acinar cells of different severity of acute pancreatitis (AP) and the release level of enzymes in vitro, and to investigate the relationship between them. Methods Two-step enzymatic digestion method was used to separate pancreatic acinar cells into 4 groups. 0. 1 μg/ml of the caerulein was added in the AP group. Caerulein and LPS (bacterial lipopolysaccharide, 10 mg/L) were added in LPS group. Caerulein and OCT (octreotide, 100 ng/ml) were added in OCT group. Medium was added in the control group. AO (acridine orange) and EB (ethidium bromide) double staining method was used to detect the incidence of apoptosis or oncosis of acinar cell. The release of intracellular enzyme was detected by measuring the concentrations of amylase and LDH in cell culture media by colorimetry method. Results The apoptosis index was 2.2 + 0.4, 6.4 ± 0.6, 4.6 + 0.4, 11.2 +1.2 in the control group, AP group, LPS group, OCT group; while the oncosis index was 3.0 +0.4, 17.2 ±1.6, 23.0 ± 2.2, 12.8 ± 1.4 in the control group, AP group, LPS group, OCT group; the release of LDH was (2180 ±240), (8060 ±930), (9460 +920), (6860 ±740) U/dl, the level of amylase was (1750 ± 190),(3820 ±460), (4420 ±480), (2260 ±260)U/L. All the values in the experiment groups were significantly higher than that in control group ( P ＜ 0.05 ). The oncosis index, LDH, amylase in LPS group was significantly higher than that in AP group ( P ＜ 0.05 ), but the apoptosis index in LPS group was significantly lower than that in AP group ( P ＜ 0.05 ). The apoptosis index in OCT group was significantly higher than that in AP group ( P ＜ 0. 05 ), but the oncosis index, LDH, amylase was significantly lower than that in AP group ( P ＜ 0. 05 ).Conclusions Induction of apoptosis and reduction of oncosis in AP pancreatic acinar cells can reduce the release of enzyme in acinar cells.

DEDD is a member of the death-effector domain protein family. DEDD inhibits the Smad3 mediated transcriptional activity and participates in the regulation of apoptosis. In this study, how the death-effector domain of DEDD participates in the regulation of Smad3 activity and apoptosis has been further investigated. Immunoblotting, immunofluorescence and immunoprecipitation had been used to detect the effects of the full length DEDD and its two truncated mutants, N-DEDD and C-DEDD on Smad3 subcellular distribution, phosphorylation, and interaction between Smad4. The effects of the full length DEDD and its two truncated mutants on cell apoptosis and proliferation had also been explored by flow cytometry and MTT assay. It showed that DEDD and N-DEDD inhibit TGF-beta1 induced Smad3 nuclear translocation and the formation of Smad3-Samd4 complex. DEDD and its two mutants can induce cell apoptosis and inhibit cell proliferation. These results suggested that DEDD inhibits the activity of Smad3 through its death-effector domain. Both the two truncated mutants of DEDD participate in the regulation of apoptosis and cell proliferation.

·AIM: To investigate the efficacy and security of the silicone membrane implant through observing intraocular pressure, filtering blebs and histopathologic results.·METHODS; Forty rabbits were randomly divided into 4 groups. Each group consisted of 10 rabbits. Trabecu-lectomy and silicone membrane implantation were performed in one eye of each rabbit, while the other eye was only performed trabeculectomy as control. The change of intraocular pressure, filtering blebs were observed postoperatively. And each eye had undergone histopathologic examination.·RESULTS:The duration of low intraocular pressure and existence of filtering blebs in implanted eyes was longer than that in controlled eyes. Light microscopy revealed that patent drainage tract and biting site could be seen in silicone membrane implanted eye. The activity and regularity of fibroblast proliferation in implanted eyes and controlled eyes were similar. Excessive expression of fibroblast proliferation was not induced by silicone membrane.·CONCLUSION: Characterized by safety, effectiveness and simplicity, silicone membrane implantation can act as a new drainage surgery.KEYWORDS: silicone membrane; drainage surgery;intraocular pressure

Objective:To investigate the value of MRI and MSCT in TNM staging of laryngocarcinoma.Method:Thirty-seven patients with laryngocarcinoma were underwent by contrast enhanced scan and muhiplanar reconstruction.Thirty-five patients with laryngocarcinoma were underwent contrast enhanced muhislice spiral CT,which of them were done by MPR.There are 28 cases which were scan by MRI and MSCT in the two former nd we contrasted the accuracy rating in laryngeal manifestation of abnormality.In the former two groups,we observed them the variability in the aspect of lymphaden metabasis and TNM staging.Result:In all 28 cases,the MRI had better accuracy rating in displaying the parts of preepiglottic space,larynx side interspace,lingual root,eck tissue,vocal cord.In the TNM staging,there was no difference in stage one in accuracy rating,as the stag stepping up,the accuracy rating of MRI had became better.The last result was that the two methods had difference in staging.In the two methods,MSCT had better sensitivity,specificity and accuracy rating.Conclusion:RI and MSCT had good accuracy rating in TNM staging,MRI has better accuracy rating in some of laryngeal,but as the lymphaden metabasis,the MSCT was better.There were variability in staging,and the MRI was better.

BACKGROUND:Shear stress can directly mediate the expression of endothelial cells, especially some cytokine genes whose codes are related to angiogenesis. Otherwise, flow shear stress of blood plays an importantly biological role in regulating vascular structure and function.OBJECTIVE: To observe the effects of laminar flow shear stress on the proliferation of vascular endothelial cells and the expression of protooncogene c-myc in human cerebral arteriovenous malformation.DESIGN: Randomized controlled study.SETTING: Department of Neurosurgery, Shenyang General Hospital of Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Neuromedical Institute, General Hospital of Shenyang Military Area Command of Chinese PLA from November 2006 to February 2007. Fresh samples of human cerebral arteriovenous malformation were derived from 20 patients who were of grade Spetzler Ⅱ -Ⅲ and received resection of human cerebral arteriovenous malformation in the Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA in 2006. All cases were diagnosed with whole-brain angiography before operation. The main reagents and equipments were detailed as follows: M199 culture media (Gilbco BRL), quality fetal bovine serum (HyClone), endothelial cell growth supplement (ECGS; Sigma, USA), CO2 incubator (Forma Scientific, USA), flow cytometry analysis of cell cycle kit (BD Company), flow cytometer (FACS Calibur, BD Company), rat-anti-human c-myc monoclonal antibody (Santa Cruz Company, USA), and reverse transcription polymerase chain reaction (RT-PCR) kit (Promega).METHODS: Tissue explants adherent method was used to culture vascular endothelial cells of human cerebral arteriovenous malformation, and then the cells were classified into 4 groups based on degree of shear stress, including control group, low shear stress group, moderate shear stress group and high shear stress group. Cultured endothelial cells of human cerebral arteriovenous malformation were put in a parallel plate flow chamber. In addition, cells in the low,moderate and high shear stress groups were stressed by low, moderate and high shear stress for 8 hours, respectively.However, shear stress in the control group was 0 Pa. Flow cytometry was used to measure proliferation index, and the expression of c-myc protein and c-myc mRNA were determined by immnocytochemistry and RT-PCR analysis respectively.MAIN OUTCOME MEASURES: Expressions of c-myc protein and c-myc mRNA and proliferation index in endothelial cells under various degrees of shear stress.RESULTS: ① Proliferation index: Proliferation index was higher in the moderate and high shear stress groups than that in the control group (P ＜ 0.05, 0.01). ② Expression of c-myc protein: Immuneposjtjve expression of c-myc protein was gradually increased with the increase of shear stress and there were significant differences in the three shear stress groups as compared with control group (P ＜ 0.05-0.01). ③ Expression of c-myc mRNA: Proliferation index of endothelial cells was higher in the low and moderate shear stress groups than that in the control group (P ＜ 0.05).CONCLUSION: Flow shear stress can induce expression of c-myc and activate expression of c-myc gene based on gene transcription so as to promote the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation