Objective To examine the effects of intracerebroventricular administration of adrenomedullin (AM) on the expression of Fos and spontaneous electric activity of area postrema (AP) neurons in sino-aortic denervated rats. Methods To determine the expression of Fos and the spontaneous electrical activity of AP neurons in male Sprague-Dawley rats by immunohistochemistry. Results Following Intracerebroventricular administration of AM (1 nmol/kg, 3 nmol/kg), Fos-like immunoreactive (Fos-LI) neurons and the discharge rate of AP neurons markedly increased. Pretreatment with calcitonin gene-related peptide(CGRP) receptor antagonist CGRP8-37 (30 nmol/kg) significantly inhibited the effects of AM (3 nmol/kg). Conclusion AM may activate the neurons in AP via CGRP receptors.

Objective Discoidin domain receptor 2 is a kind of receptor tyrosine kinases, which was found to be over-expressed in synovial fibroblasts in rheumatoid arthritis (RA). The present study was to construct and express wild type (FLDDR2), Fc chimera (FcDDR2) and truncated form of mouse discoidin domain receptor 2 (ttDDR2) for further study. Methods Full-length, truncated form of mouse DDR2 were amplified by RT-PCR. A chimera form of mouse DDR2 was constructed by replacing part of the extracellular domain with Fc fragment of human immunoglobulin G1 (IgG1). Eukaryotic expression vectors of the different forms of mouse DDR2 were constructed by subcloning the PCR products into pcDNA3.1(+) or pMKIT-Neo. Then COS-7 cells were transient transfected with the eukaryotic expression vectors of full-length (FLDDR2), truncated (ttDDR2) and chimeric form of mouse DDR2 (FcDDR2). Successful transfection and expression was confirmed by Western blot(WB) and Immunoprecipitation (IP)/WB. With or without stimulation with soluble type I collagen for 3h, phosphorylation level of transfected cells were detected by IP/WB. Eukaryotic expression vectors of full-length, truncated form and chimeric forms of mouse DDR2 were successfully constructed and confirmed by sequencing. After transient transfection, the expression of these three forms in the respectively transfected cells was observed by Western blot. Results The result of IP/WB suggested that the chimeric form of mouse DDR2(FcDDR2) could be properly expressed in the COS-7 cells. Under the condition of collagen, decreased tyrosine phosphorylation of FLDDR2 was detected in the COS-7 cells that were cotransfected with ttDDR2 and FLDDR2, comparing with that in COS-7 cells transected with only FLDDR2. There was no obvious difference in phosphorylation level between FcDDR2 without collagen stimulation and FLDDR2 with the collagen stimulation. Conclusion Three different forms of mouse DDR2 were successfully constructed. FcDDR2 could be an activator for its auto-phosphorylation. And ttDDR2 could be a partially negative competitor.

We examined the effects of intracerebroventricular ( i. c. v) administration of adrenomedullin (ADM) on catecholaminergic neurons and the expression of c-fos gene in rat brain nuclei involved in cardiovascular regulation using double immunohistochemical method for Fos and tyrosine hydroxylase (TH). The results showed that: ( 1 ) Following icy administration of ADM (3 nmol/kg) , double-labeled neurons for Fos and TH were significantly increased in the area postrema ( AP), the nucleus of the solitary tract ( NTS), the nucleus paragigantocelluaris laterialis (PGL) and the locus coeruleus (LC). (2) Pretreatment with calcitonin gene-related peptide receptor antagonis CGRP8-37 (30 nmol/kg) significantly reduced the action of ADM (3 nmol/kg) in the brain. The present study suggested that ADM might activate the neurons of the brain nuclei involved in cardiovascular regulation, and supported the hypothesis that the central action of ADM were induced by activating the catecholaminergic neurons of brainstem nuclei involved in cardiovascular regulation, CGRP receptor might mediate the effects of ADM.

Objective To investigate oxidative damage effect of the serum of severe preeclamptic patients on human umbilical vein endothelial cells (HUVEC).Methods (1) HUVEC were randomly divided into 4 groups according to the following:blank group as control,normal group added 20％ normal sera of pregnant women,group PE added 20％ sera of severe preeclamptic patients,and group PE + Cat added 20％ sera of severe preeclamptic patients plus 3 x 103 U/ml catalase.After cultured for 24 hours,the injury morphology and APO2.7 expression of HUVEC were detected by transmission electron microscopy and flow cytometry respectively.(2) Under the real-time scanning by laser scanning confocal microscopy,HUVEC were randomly divided into 4 groups according to the following:control group added 100 μmol/L H2O2 as positive control,normal group,group PE,and group PE + Cat.HUVEC of each group was scanned for 120 seconds to determine levels of reactive oxidative species (ROS),calcium homeostasis,and mitochondria membrane potential.Results (1) Obvious injury morphology of HUVEC was observed in group PE,and it was obviously improved by catalase in group PE + Cat.Percentage of HUVEC expressed APO2.7 was (37.8 ± 1.1) ％ in group PE,which was significantly higher than (13.4 ± 1.1) ％ in blank group or (13.5 ± 1.5) ％ in normal group,but significantly lower than (19.2 ± 1.6) ％ in group PE + Cat (all P ＜ 0.01).(2) The fluorescence intensity curves of intracellular ROS and calcium showed slowly rising in group PE,but no obvious changes in normal group and PE + Cat.The values of ROS and calcium in group PE (12.0±1.3,4.1 ±0.7) were higher than those in normal group (1.1 ±0.4,0.6 ±0.4),but lower than those in group PE + Cat (1.5 ± 0.5,0.9 ± 0.5 ; all P ＜ 0.01).Conclusion The serum of severe preeclamptic patients caused oxidative damage on HUVEC by increasing intracellular ROS generation,calcium overload,and decreasing mitochondrial membrane potential.

The pancreatic tissues from patients with islet cell hyperplasia,insulinoma,and pancreatic adenocarcinoma,as well as normal pancreatic tissues were embedded with paraffin,serial sections were cut and mounted on glass slides.Immunohistochemical staining was carried out with N-myc down-stream regulated gene 2 (Ndrg2) monoclonal antibody by means of ABC method,and Western blotting was carried out to detect the expression and distribution of Ndrg2.The results showed that Ndrg2 positive immunoreactivity was mainly localized in the cytoplasm of islet cell,being similar to the localization of insulin positive immunoreactivity.The number and volume of pancreatic islets were increased in the patients with islet cell hyperplasia,and Ndrg2 expression was also increased.Western blotting results showed that the expression of Ndrg2 in the pancreas of patients with islet cell hyperplasia was increased compared with normal group.The above results suggest that Ndrg2 may play an important role in performing physiological function of islet cells.

Objective To investigate the effect of gene silencing of Y-box binding protein-1(YB-1) by RNA interference on the proliferation and migration in prostate cancer cells lines PC-3 cells .Methods YB-1 siRNAs(pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2) were synthesized and transfected into cloned into the the PC-3 cells by liposome .The expressions of YB-1 were measure by RT-PCR and Western blotting .The proliferation and migration were respectively detected by MTT and Transwell method . Results ThemRNA and protein expressions of YB-1 were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2 (P<0 .05) ,compared with the control group ,the inhibition ratio of mRNA expression was 36 .23% and 39 .42% respectively and the inhibition ratio of protein expression was 41 .56% and 55 .33% respectively .The proliferation and migration were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2(P<0 .05) .Conclusion YB-1 gene silencing by RNA interference inhibits the proliferation and migration in prostate cancer cells lines PC-3 cells .

Objective To investigate the effect of hydrogen sulfide (H2S) on the cholesterol efflux and ATP-binding cassette transporter A1 (ABCA1) expression in foam cells .Methods RAW 264 .7 macrophages were incubated with oxidized low density lipoprotein to induce foam cells .Foam cells were incubated with H2S donor sodium hydrosulfide .Cholesterol efflux from macropha-ges was tested by labed cholesterol .The cellular levels of free cholesterol (FC) ,cholesterol ester (CE) and total cholesterol (TC) were measured by high performance liquid chromatography assays .The mRNA and protein expressions of ABCA1 were detected by Real-time PCR and Western blot .Results Compared with the foam cells ,the rates of cholesterol efflux were significantly in-creased ,the levels of TC ,FC ,CE and CE/TC ratio were significantly decreased(P<0 .05) and expression of ABCA1 was signifi-cantly increased by treatment with H2S in dose-and time-dependent manner(P<0 .05) .Conclusion H2S up-regulates of expres-sion ABCA1 and promotes cholesterol efflux in RAW 264 .7 macrophage-derived foam cells .

Besides respiratory system,Mycoplasma pneumoniae infection can also cause diseases in other organ systems,and nervous system impairment is the common extrapulmonary complications. Recent years,the morbidity of Mycoplasma pneumoniae encephalitis is uprising. As the pathogenesis of Mycoplasma pneumonia encephalitis is not clear,so uniform diagnosis standard hasn't been acquired up to now. Its treatment mainly uses macrolides with less na-gative effects on children's body growth,and intravenous immunoglobulin could also be used jointly. The prognosis of Mycoplasma pneumonia encephalitis is related with clinical manifestation,age of onset,imageological change and whe-ther receiving timely treatment or not.

BACKGROUND:Increasing evidence col ected from animal experiments or laboratories shows that bone mesenchymal stem cel s possess potent immunosuppression and anti-inflammation effects and cartilage regenerative capability. The microenvironment in human knee joint of osteoporosis is more complex and involves lots of bioactive factors and immunologic mechanisms. OBJECTIVE:The analyze the therapeutic effects of intra-articular injection of autologous bone marrow mesenchymal stem cel s on mild-to-moderate osteoporosis, and to investigate the mechanisms for anti-inflammation, immunoregulation and reversion of cartilage degradation. METHODS:About 15 mL bone marrow was aspired from 26 patients with mild-to-moderate osteoporosis and taken to the laboratory where bone marrow mesenchymal stem cel s were isolated and characterized in terms of some surface markers by a flow cytometer, and the ability of osteogenic and adipogenic differentiation was analyzed. The bilateral knees of each patient were divided into two groups at random. The experiment group were treated with autologous bone marrow mesenchymal stem cel s by intra-articular injection (2×107 cel s), while the control group injected the same volume of control medium without bone marrow mesenchymal stem cel s. Western Ontario and McMaster Universities Osteoarthritis Index was used to estimate the function of the knee joints before and after the treatment. We col ected the joint fluid before, 2 and 4 weeks post treatment, and then measured the production of interleukin-1, interleukin-10, tumor necrosis factor-α, cartilage oligomeric matrix protein using ELISA. RESULTS AND CONCLUSION:According to the standardized culture-expansion protocol, a sufficient number of bone marrow mesenchymal stem cel s (more than 2×107 passage 3 cel s) were obtained for intra-articular injection. The bone marrow mesenchymal stem cel s isolated from patients were positive for CD105, CD29 and negative for CD45, CD34, and had the fair osteogenic and adipogenic capacities. The function of the knee joints was improved obviously after treatment. In the experimental group, the secretion of both interleukin-1, tumor necrosis factor-αand cartilage oligomeric matrix protein in the joint fluid was suppressed, while the interleukin-10 level was up-regulated compared to the control group. The results suggest that intra-articular injection of autologous bone marrow mesenchymal stem cel s can exert good effect in mild-to-moderate osteoporosis patients. Furthermore, bone marrow mesenchymal stem cel s via intra-articular injection can function through anti-inflammation and reversion of degradation of the articular cartilage, which is a new promising approach for treating mild-to-moderate osteoarthritis.

Objective To study the effect of electrical stimulation breath training on lung function of patients following pulmonary lobectomy. Methods 62 patients following pulmonary lobectomy were randomly allocated into experimental group (n=30 )and control group (n=32). The experimental group received a 4-week supervised electrical stimulation breath training program using an electric stimulus feedback trainer (20mins per time, 3 times per week);The control group received postoperative routine nursing. Cadiopulmonary function evaluation of 2 groups were tested before and after the experiment. The evaluation included the 6-min walking test (6MWD), FVC, FEV1,W,AT and VO2max/kg. Results After 4 week training, the value of 6MWD,W,FVC,FEV1 all improved, compared to the baseline value (P < 0.05) and the value of 6MWD,W,FVC,FEV1 were more obvious in experimental group, compared to control group(P<0.05). The AT value and the VO2max/kg value increased than the baseline value (P<0.05)and the improvement degree was more remarkable in experimental group than that in control group (P<0.05). Conclusion Electrical stimulation breath training can improve cardiopulmonary function of the patients following pulmonary lobectomy.