Objective To evaluate the efficacy and safety of endoscopic mucosal resection (EMR) for rectal carcinoid tumors. Methods From January 2006 to January 2009, EMR was performed in 28 patients with rectal carcinoids, who were followed up to evaluate the therapeutic effect and safety. Results Tumor diameters varied from 0.4 cm to 1. 2 cm (mean 0.7± 0. 2 cm). Negative resection margin was a-chieved in 26 cases (92. 9% ), tumor margin within 0. 1 cm of resection margin in 1 (7. 1% ) , and two margins coincided in 1 patient (7. 1% ). Hemostasis was performed with metal clips in 14 patients (50% ) and argon plasma coagulation (APC) in 9 (32. 1% ). Except for rectal bleeding in 1 patient (3. 6% ) , no other complications were observed. There was no recurrence in any patients during a follow-up of 6-36 months. Conclusion EMR is a useful and safe method for treatment of small rectal carcinoid tumor which does not cross submucosal layer.

Objectives: To study and prepare monoclonal antibody against glycophorin A of human erythrocyte(GPA McAb).This antibody is a key reagent in preparation of bispecific antibodies for rapid whole-blood immunoassay.　Methods: BALB/c mice received GPA antigen injection. Hybridoma was produced by traditional techniques. Hybridoma was determined with ELISA and indirect agglutination(IA) method.　Results: All 3 GPA McAb reacted with GPA, and they did not autoagglutenate with four types of red cells(type A,B,O,AB).Of the 3 McAb-GPA, two antibodies were IgG1,one IgG2 subtypes. Two IgG1 McAb-GPA can be used in making bispecific antibody in preparation of rapid whole-blood immunoassay.　Conclusions: Immunogenity of GPA was enhanced after coupling with the native serum albumin of the immunized animal, and high titer GPA McAb could be easily obtained. This result is important in making bispecific antibodies in preparation of rapid whole-blood immunoassay.

Background Growth factor-induced proliferation and migration of retinal pigment epithelium (RPE) cells are the major pathological changes of proliferative vitreoretinopathy (PVR).Arsenic trioxide ( As2O3 ) is an active ingredient of Chinese traditional medicines,which has an inhibition on proliferation and migration of tumor cells.However,it is not clear whether As2O3 could inhibit growth factor-induced proliferation and migration of RPE cells. Objective This study was to explore the effects of As2O3 on epidermal growth factor (EGF)-induced proliferation and migration of ARPE-19 cells. Methods RPE cell line (ARPE-19 cells) were cultured.Different concentrations of As2O3(0,0.5,1.0,2.0,5.0,10.0,20.0 μmol/L) were added in the culture plate to treat ARPE-19 cells with or without 10 mg/L EGF in serum-free group for 24 and 48 hours,respectively.The MTT colorimetric assay was used to check the cell viability and evaluate the drug toxicity.The effects of As2O3 on EGF-induced proliferation of ARPE-19 cells were analyzed to get an effective and avirulent concentrations of As2O3.The effects of As2O3 on EGF-induced migration of ARPE-19 cells were observed by scratch-wound assay and the Boyden chamber assay.Results MTT assay showed that the A values were gradually declined with the increase of As2O3 concentrations after As2O3 treatment without EGF for 24 hours and 48 hours ( Fgroup =38.269,P =0.000 ; Ftime =0.874,P =0.358 ).Compared with the control group,no significant differences were seen in the A values of ARPE-19 cells in 0.5-5.0 μmol/L groups (all P＞0.05).Meantime,As2O3 reduced the A values of ARPE-19 cell with 10 mg/L EGF in dose- and time-dependent manner ( Fgroup =152.155,P =0.000 ; Ftime =51.649,P =0.000 ).There were not significant differences in 10 mg/L EGF-induced cell growth after 0.5,1.0,2.0 μmol/L As2O3 was added for 24 and 48 hours ( Fgroup =2.215,P =0.126 ;Ftime =2.230,P =0.155).However,when 5.0-20.0 μmol/L As2O3 added,the A values of 10 mg/L EGF-induced ARPE-19 cells lowed,showing a significant difference in comparison with the control groups ( all P＜0.05),with the cellular inhibiting rate 12％,32％,37％ in 24 hours and 39％,44％ and 53％ in 48 hours.Scratch-wound assay showed that EGF-induced horizontal migration of ARPE-19 cells was slow after 0.5-2.0 μmol/L As2O3 treated,and the same results also appeared in cell lognitudinal migration by Boyden chamber assay,with the inhibitory rates 22％,33％ and 46％ respectively. Conclusions As2O3 is avirulent on ARPE-19 cells within definite concentration range.At ≤ 2.0 μmol/L concentrations,As2O3 dose not affect EGF-induced proliferation of ARPE-19 cells,but it suppresses EGF-induced cell migration.At ≥ 5.0 μmol/L concentrations,As2O3 plays an inhibitory role to EGF-induced proliferation of ARPE-19 cells.

To prove the influence of protein isoaspartyl-methyltransferase ( PIMT ) on the cell apoptosis of fibroblast-like synoviocytes of RA.Methods: The expression vector of PIMT was constructed and transfected in to the RA-FLS, the impact of PIMT on the cell apoptosis of RA-FLS was observed by overexpressing the vector of PIMT.Results:The mRNA and protein level of PIMT in RA-FLS was increased after transfected the vector of PIMT into RA-FLS;compared with the normal cultured RA-FLS and the RA-FLS transfected with the empty vectors ,the cell apoptosis level was also increased.Conclusion:The decreased expression level of PIMT in RA-FLS is an important reason for reduce apoptosis of RA-FLS,and PIMT can affect the imbalance of proliferation/ap-optosis in the RA-FLS.

Background Oxidative damage plays an important role in pathogenesis of age-related macular degeneration( AMD ),and its mechanism is the destroy of blood-retinal barrier.Müller cells is a primary component to stabilize the inner barrier of the blood-retina.Researches showed that epidermal growth factor(EGF) can promote the proliferation and migration of animal Müller cells,but less study was found in the effect of EGF on human Müller cells. Objective The present study was to investigate the effects of EGF on the proliferation and migration of human Müller cells and its molecular mechanism. Methods Human Müller cell line MIO-M1 cells were cultured and incubated,and cultured cells were identified using glial fibrillory acidic protein (GFAP),factor Ⅷ,α-smooth muscle actin( α-SMA ),keratin and S-100.Different concentrations of EGF( 0,1,10,30,100 mg/L)was added in freeserum DMEM,and the positive rate of the cells was calculated using 5-bromo-2-deoxyuridine(BrdU) method.The cells were divided into EGF group,H2 O2 group,EGF + H2 O2 group,glucose oxidase ( GO ) group,GO + EGF group,EGF + LY294002+H2O2 group according to the different intervention,and the effects of LY294002 on the proliferation of Müller cells (A590 )were detected by colorimetric assay for cellular growth and survival( MTT assay).The scratch test of Müller cells was used to assess the influence of EGF(0,1,10,30,100 mg/L)on H2 O2-induced damage of human Müller cell.Western blot was used to detect the cell proliferation under the protection of EGF on co-cultured cells using LY294002 and H2O2 and the activation of Akt signal pathways. Results The proliferative rates of the cells were 28.0％,32.9％,39.0％ in 10,30,100 mg/L EGF groups respectively and obviously higher than those in 0,1 mg/L EGF groups (24.5 ％,26.2 ％ ).Under the H2O2 culture,GO culture,respectively,the A570 value of the Müller cell in high concentrations of EGF groups was significantly increased in comparison with lower concentrations EGF groups with the statistical significance among the groups( F=23.582,P=0.000).Compared with EGF+H2O2 group,the A570value of the Müller cells was lowed in EGF+LY294002+H2O2 group.The maximum migration rate of Müller cells was found in 10 mg/L EGF group.Western blot revealed that the presence of H2O2 reinforced the expression of Akt in Müller cells,however,pretreatment with 100 mg/L EGF antagonized the harmful effect of H2O2 on Müller cells.Meanwhile,pretreatment with EGF and LY294002 reduced the expression of Akt in Müller cells. Conclusions EGF can induce the proliferation and migration of human Müller cells with the strongest effect in 10 mg/L.100 mg/L exogenous EGF has a stronger protection to the Müiller cells against H2O2-induced cell damage by activating the PI3KAkt cell survival pathway.

Objective Diabetic mice models were established.Hearts were separated from the body,reperfused after ischemia,and disposed of with the extraneous calcitonin gene-ralated peptide (CGRP),to prove if the CGRP had protective effect on the early-staged diabetic mice with cardiac ischemia.Method ICR mice were injected with Streptozocins intraperitoneally to establish diabetic model.The model mice and the normal mice were randomly divided into 4 groups:two diabetic groups,two control groups.Hearts of the experimental mice were taken out from the chest cavity alive,and hanged on the Langendorff steadily for 30min.Western blot and radioimmunoassay techniques were used to test expression of Vanilloid receptor(VR1) and CGRP in myocardic tissue;Madlab system was used to test the cardiac function,which including left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP),heart rate (HR),and coronary artery flow (CF),in the process of ischemic reperfusion,And ELISA assay kit was used to measure the concentration of the lactate dehydrogenase (LDH) in perfusion fluid collected from the heart.Result Expressions of VR1 and CGRP in diabetic hearts were significantly lower than those in normal ones 2 weeks later(P＜0.01).By comparing with normal hearts,diabetic hearts had higher LVEDP and CF (P＜0.05),and lower LVDP and HR(P＜0.05).However,release of LDH were lower than normal ones (P＜0.05).Predisposition of normal and diabetic hearts with CGRP can improve the cardiac function after ischemie injury.And the beneficial effect was more profound in early-staged diabetic hearts than in normal ones.Conclusion The diabetes disease(DM) can impair the expression of CGRP in myocardiac tissue.The extraneous CGRP may exert more potent protective effect on cardiac function in the early-staged diabetic heart.

Human amylin (hAmylin) is co-released with insulin from pancreatic B-cells and the actions of this peptide on its target tissues maintain the cell excitability and glucose homeostasis. Inappropriate control of hAmylin secretion may result in human disease, particularly Alzheimer's disease (AD). It's unknown that which kind of receptor is activated by human amylin, leading to the neurotoxicity in neurons of brain. Nicotinic acetylcholine receptors (nAChRs) are known to play a critical role in a variety of nervous diseases. In the present study, we sought to determine the inter-relationships between these two receptors by examining the actions of hAmylin and nicotine on whole-cell currents and membrane potential in basal forebrain neurons. Whole cell patch-clamp recordings were performed on enzymatically dissociated neurons of the diagonal band of Broca (DBB), a cholinergic basal forebrain nucleus. The results showed that either hAmylin or nicotine individually caused a dose-dependent (1 nmol/L-20 µmol/L) membrane depolarization and an increase in firing frequency of DBB neurons. Application of AC253, an amylin receptor antagonist, blocked the excitatory effects of not only hAmylin but also nicotine; dihydro-β-erythroidine (DHβE), a nAChR antagonist, also blocked the effects of nicotine and hAmylin. These electrophysiological results suggest that hAmylin receptor and nAChRs on DBB neurons are coupled and may function in a co-operative manner to influence the excitability of DBB neurons. This finding is important for us to understand the cause and mechanisms of AD.
Animals ; Brain ; metabolism ; physiology ; Diagonal Band of Broca ; metabolism ; physiology ; Humans ; Islet Amyloid Polypeptide ; pharmacology ; Male ; Neurons ; metabolism ; physiology ; Nicotine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Islet Amyloid Polypeptide ; physiology ; Receptors, Nicotinic ; physiology

This work was mainly studied the effects of the four alkaloids from Coptidis Rhizoma on the mouse peritoneal macrophages in vitro and preliminarily discussed the regulating mechanisms. The effect of alkaloids from Coptidis Rhizoma on the vitality of macrophages was measured by the MTT assay. The effect of alkaloids on the phagocytosis of macrophages was determined by neutral red trial and respiratory burst activity was tested by NBT. The expressions of respiratory-burst-associated genes influenced by alkaloids were detected by qRT-PCR. The conformation change of membrane protein in macrophages by the impact of alkaloids was studied by fluorospectro-photometer. Results showed that the four alkaloids from Coptidis Rhizoma could increase the phagocytosis of macrophages in different level and berberine had the best effect. Berberine, coptisine and palmatine had up-regulation effects on respiratory burst activity of mouse peritoneal macrophages stimulated by PMA and regulatory activity on the mRNA expression of PKC, p40phox or p47phox, whereas the epiberberine had no significant influence on respiratory burst. Moreover, alkaloids from Coptidis Rhizoma could change the conformation of membrane protein and the berberine showed the strongest activity. The results suggested that the four alkaloids from Coptidis Rhizoma might activate macrophages through changing the conformation of membrane protein of macrophages and then enhanced the phagocytosis and respiratory burst activity of macrophages. Furthermore, the regulatory mechanism of alkaloids on the respiratory burst activity of macrophages may be also related to the expression level of PKC, p40phox and p47phox.
Alkaloids ; pharmacology ; Animals ; Cells, Cultured ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression ; drug effects ; Macrophages, Peritoneal ; drug effects ; Mice ; Phosphoproteins ; genetics ; metabolism ; Protein Kinase C ; genetics ; metabolism ; Rhizome ; chemistry

Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro.
Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000μεrespectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection.
Results After being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues (all P<0.05). DNA ladders were seen in the bone tissue under 3000 and 4000μεmechanical load. Moreover, there was significant enhancement in the activities of caspase-3/8/9 in the mechanical stress group of 3000 and 4000με(all P<0.05).
Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

OBJECTIVETo investigate the biodistribution of (18)-NaF as an imaging agent for position emission tomography (PET) in rat models of osteoporosis.

METHODSOsteoporosis was induced in 10 rats via injection with an excess of dexamethasone phosphate sodium, and the biodistribution of (18)-NaF in the rats was studied, with another 10 normal rats as the control group. (18)-NaF PET was also performed in 8 healthy volunteers, and the uptakes of (18)-F- in the bone tissues were measured.

RESULTSCompared with the control rats, the osteoporotic rats showed significantly decreased (18)-F- uptake, especially in the femoral neck, lumbar vertebrae, the 7th rib and the tibia (P<0.05). Dynamic chest PET scanning in the volunteers revealed obvious (18)-F- uptake in the spine, ribs and humerus 20 s after injection of the imaging agent. (18)-F- uptake significantly increased with time in the bones, reaching the peak level 60 min after the injection, and whole-body PET at this point demonstrated obvious skeletal (18)-F- uptake, with high skeletal-to-muscle (STM) ratio that averaged 8.12.

CONCLUSION(18)-NaF is an excellent skeletal imaging agent for clinical skeletal blood flow and metabolism measurements. The uptake of (18)-NaF has significant difference between normal and osteoporotic bone tissues, indicating the value of (18)-NaF PET for study of osteoporosis.

Adult ; Animals ; Bone and Bones ; diagnostic imaging ; metabolism ; Female ; Fluorine Radioisotopes ; pharmacokinetics ; Humans ; Male ; Osteoporosis ; diagnostic imaging ; Positron-Emission Tomography ; methods ; Radiopharmaceuticals ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; pharmacokinetics ; Tissue Distribution