Objective:To establish peri-implantitis model in beagle dog. Methods:Mandibular first, second and third premolar and first morlar of 5 beagle dogs were extracted. 3 months after tooth extraction,6 branemark pure titanium implants were bilaterally inserted into the posterior mandible in each dog. After healing-abutment connection, cotton ligatures were placed around the healing-abutments and then plaque was accumulated. Clinical and radiographic recording were repeated 1,3 and 5 months respectively after baseline data had been obtained. Surgical recording and pathological changes of peri-implant bone morphology were studied. Results:1,3 and 5 months after the formation of ligature-induced peri-implant inflammation, plaque index,pocket depth and gingival index were singnificantly increased(P

Objective To observe the changes of HMGB-1 of burned patient with sepsis and to investigate the effect of carbaehol on production of HMGB-1 from human monocytes stimulated by lipopolysaccharide(LPS) and its mechanism. Methods The peripheral blood samples of burned patients with sepsis and healthy donors were taken for isolation of monoeyte. The subjects were divided into six groups: the menocytes group were added only with 1640 culture medium,LPS group were added only with LPS;nicotine group and carbachol group were with carbachol first or nicotine for 5 rain and then with LPS. The levels of HMGB-1 were tested by EIJSA. α-bungarotoxin + carba-chol group were added with atropine or α-bungarotoxin for 5 min and then were given carbachol and 5 min later were stimulated with LPS. 48 h-incubation later supemate were collected for the detection of HMGB-1 by ELISA. Results HMGB-1 levels of burned patients with sepsis were ( 12.94±6.54)μ/L,which were much higher than that of healthy donors( [ (2.01±0.03 )μ/L), P <0.01 ]. When monoeytes were stimulated by LPS alone, concentrations of HMGB-1 were (9.39±1.37 ) μ/L, which were obviously higher than that of controls [( 1.48±0.69 ) μ/L), P < 0.01]. Mter pretreated by carbaehol or nicotine,concentrations of HMGB-1 were (3.52±1.64)μ/L and (4.01± 1.56) μ/L respectively, which were obviously decreased compared with that of LPS stimulation alone group ( P < 0.01). When pretreatedby atropine before addition of carbachol, there was no significant changes in concentrations of HMGB-1 [ (3.87±2.01 )μ/L]. When pretreated with α-Bungaretoxin before carbachol adminitration, inhibitive effect of carbachol on production of HMGB-1 were blocked [(8.97±1.97 )μ/L ]. Conclusion The release of HMGB-1 in burned patients with sepsis is increased. Carbachol could obviously reduce the production of HMGB-1, which may affect through activating α 7 subunit of cholinergic N receptor.

Objective To explore the association between mild cognitive impairment (MCI) and hypersensitive C reactive protein (hs-CRP),interleukin-1β (IL-1β) and interleukin-6 (IL-6) among Uygurs and Hans in Xinjiang region,China.Methods From July 2008 to October 2010,the epidemiological investigation was performed in Southern,Eastern and Northern of Xinjiang.Based on the diagnostic standard of United States psychiatric society of spirit obstacles diagnosis and statistics manual Ⅳ amendment version in the mild cognitive function,483 MCI patients were diagnosed.Finally,314 MCI patients were selected from above according to the completion of data.Moreover,299 subjects were randomly selected as the control group from the investigation.General information and fasting plasma were collected,and blood glucose,blood lipid and biochemical indexes,serum hs-CRP,IL-1β,IL-6 of concentration were tested.The association between MCI and hs-CRP,IL-1β and IL-6 were analyzed with SPSS 17.0 software.Results (1) The concentrations of serum hs-CRP,IL-1β and IL-6 in MCI group (3.40 (6.53) mg/L,0.09 (0.09) ng/L,136.08(96.77) pg/L) were significantly higher than that in control group (2.99 (3.91) mg/L,0.07(0.06) ng/L,79.32(68.79) pg/L) respectively (Z =-2.525,-2.946,-9.361,all P ＜0.05).(2)The concentrations of serum hs-CRP,IL-1β,IL-6 in Han MCI patients were significantly higher than that in Han non-MCI subject; The concentrations of serum IL-1β,IL-6 in Uygur MCI patients were significantly higher than that in Uygur non-MCI subjects; However,the hs-CRP concentration between MCI and non-MCI group is not statistically different among Uygurs.(3)Non-conditional Logistic regression analysis showed that serum IL-1β(OR =1.008,95％ CI0.897-1.071,P =0.006),hs-CRP (OR =1.096,95％ CI1.056-1.137,P =0.000),IL-6 (OR =1.011,95％ CI1.008-1.014,P =0.000) were associated with MCI.Conclusion The hs-CRP,IL-1β and IL-6 were independent risk factors for MCI.

Objective To design T-shaped shoes with a chuck regulator to facilitate the orthopedics patient in lower limb immobilization,nursing and rehabilitation training.Methods Each of the shoes was composed of the body of low temperature thermoplasticized plate,a regulator,two cross bars controlled by the regulator and a circular silica gel liner at the heel.The stability of triangles kept the limb involved at the middle position,and prevented the lower limb from inward turning,outward turning or dropping.The shape of the triangles was modified by adjusting the regulator to form an individualized fixation posture for each patient.Results The shoes facilitated clinical nursing,enhanced the patient comfort and decreased the complications.Conclusion The shoes gains advantages in wearing,low cost and repeated use,and thus is worthy promoting practically.

Objective To explore acute cerebral vascular accident factors in patients with nosocomial infection. Methods Clinical data with 680 cases with acute cerebral vascular accident were retrospectively analyzed,and the patients with hospital flu infected as the observation group, selected in accordance with 1 ∶ 1 over the same period without cerebrovascular accident combined hospital patients feel as control group. The patient age,gender,state of consciousness , invasive operation, dehydrating agent application time, whether use of antibiotics and other differences were compared . Results In 680 cases of patients with acute cerebral vascular accident, there were 90 cases of hospital infection; two groups gender, dehydrating agent application time had no difference(P ＞ 0. 05 ); the observation group compared with the control group older, unconscious, to implement invasive operation to prevent high proportion of antibiotics(P ＜0. 05), hospital infection-related factors. Conclusion Acute cerebral vascular accident patients should have a higher incidence of nosocomial infection, and be related with many factors. Taking corresponding measures against the relevant factors could prevent hospital infection.

This study investigated the modulatory effect of synthetic cannabinoids WIN55,212-2 on 5-HT(3) receptor-activated currents (I(5-HT3)) in cultured rat trigeminal ganglion (TG) neurons using whole-cell patch clamp technique. The results showed that: (1) The majority of examined neurons (78.70%) were sensitive to 5-HT (3-300 μmol/L). 5-HT induced inward currents in a concentration-dependent manner and the currents were blocked by ICS 205-930 (1 μmol/L), a selective antagonist of the 5-HT(3) receptor; (2) Pre-application of WIN55,212-2 (0.01-1 μmol/L) significantly inhibited I(5-HT3) reversibly in concentration-dependent and voltage-independent manners. The concentration-response curve of 5-HT(3) receptor was shifted downward by WIN55,212-2 without any change of the threshold value. The EC(50) values of two curves were very close (17.5±4.5) μmol/L vs. (15.2±4.5) μmol/L and WIN55,212-2 decreased the maximal amplitude of I(5-HT3) by (48.65±4.15)%; (3) Neither AM281, a selective CB1 receptor antagonist, nor AM630, a selective CB2 receptor antagonist reversed the inhibition of I(5-HT3) by WIN55,212-2; (4) When WIN55,212-2 was given from 15 to 120 s before 5-HT application, inhibitory effect was gradually increased and the maximal inhibition took place at 90 s, and the inhibition remained at the same level after 90 s. We are led to concluded that-WIN55,212-2 inhibited I(5-HT3) significantly and neither CB1 receptor antagonist nor CB2 receptor antagonist could reverse the inhibition of I(5-HT3) by WIN55,212-2. Moreover, WIN55,212-2 is not an open channel blocker (OCB) of 5-HT(3) receptor. WIN55,212-2 significantly inhibited 5-HT-activated currents in a non-competitive manner. The inhibition of I(5-HT3) by WIN55,212-2 is probably new one of peripheral analgesic mechanisms of WIN55,212-2, but the mechanism by which WIN55,212-2 inhibits I(5-HT3) warrants further investigation.

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection

Human adenovirus type 41 (HAdV-41) is considered to be a "fastidious adenovirus". E1-deleted HAdV-41 cannot be rescued or amplified in 293 cells. To propagate recombinant HAdV-41 in 293 cells, the backbone plasmid pAdbone41 was reconstructed. That is, the E3 coding sequence of HAdV-41 was deleted and replaced with the HAdV-5 E4orf6 gene; and the E1A enhancer of HAdV-5 was inserted upstream of the E4 promoter of HAdV-41. Novel adenoviral plasmid pAd41E4EE-GFP was generated by homologous recombination of the shuttle plasmid pSh41-GFP with the modified backbone plasmid in the Escherichia coli BJ5183 strain. Adenovirus HAdV-41-E4EE-GFP was rescued by transfecting 293 cells with linearized pAd41E4EE-GFP. After seven rounds of propagation, viruses were purified by the CsCl ultracentrifugation method. HAdV-41-E4EE-GFP in 1.0 ml with a particle titer of 8 x 10(10) vp/mL was obtained which had a particle-to-infectious ratio of 50 : 1. The genome of HAdV-41-E4EE-GFP was confirmed by restriction analyses and polymerase chain reaction. These results showed that a novel HAdV-41 vector system was established in which recombinant HAdV-41 could be constructed and packaged in 293 cells.
Adenoviruses, Human ; genetics ; growth & development ; physiology ; Cell Line ; Genetic Vectors ; genetics ; physiology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; metabolism ; Recombination, Genetic ; Transfection ; Virus Assembly

BACKGROUND: Electrical activity of nerve cells is based on the ion channel activity on cell membrane. Epilepsy is basically characterized by abnormal neuronal discharge. The foundation is ion channel activation on cell membrane and ion transmembrane movement, however, whether Ca2+-activated K+ channel involves in epilepsy induced by Coriaria Lactone is still unclear.OBJECTIVE: Considering rat hippocampal pyramidal neurons as target,we investigate the effect of Coriaria Lactone on neuronal Ca2+-activated K+ channels in epilepsy induced by Coriaria Lactone.DESIGN: Randomized controlled experimental trials.SETTING: Department of Neurology, West China Hospital, Sichuan University and Institute. of Myocardium Electrophysiology of Luzhou Medical College.MATERIALS: This experiment was carried out in Luzhou Medical College, Sichuan Province, from May to December 2000. Totally 100 Wistar infant rats within 24-hour ages were selected.METHODS: Wistar infant rats were anaesthetized and its hippocampus was obtained under disinfected state, pyramidal neurons were cultured for 7-10 days, neurons growing well with typical shape model were colleted normal control group, 19 dishes were added with DMEM culture medium,given different membrabe voltage and then followed by adding in te3 subgroups with 8 dishes each one. Added seperately DMEM culture medium containing f0-8, 10-7, 10~ mol/L concentration of calcium ion, and 2.0 mL/L Coriaria Lactone induced epilepsy group: added with DMEM culture medium with different dosages of Coriaria Lactone and finally tetraethylamine in each concentration of 26 dishes for totally 130 dishes.Cell-attache method and inside-out method of patch-clamp technique were used to record the neuronal single channel electricity. The open probability, average opening hour and closing hour, electric current amplitude of channel were analyzed.activated K+ channels of pyramidal neurons at normal, various membrane To observe and record the influence of Coriaria Lactone on the activation of pyramidal neuronal cell membrane, as well as the role of tetraethylamine.were only small amount of pyramidal neurons randomly opening its Ca2+-activated K+ channels and it displayed obvious voltage-dependent property.The channel electric conductance was (122.79±21.68) pS. The channels the inside-out condition, Ca2+-activated K+ channel displayed calcium iondependent property. The average opening rate was 0.022±0.006, 0.040±0.007, 0.142±0.049 when the calcium concentration was 10-8, 10-7,aria Lactone could increase the opening rate of Ca2+-activated K+ channels when the free calcium ion in bath solution was 10-8 mol/L and memLactone, 1.0 mL/L Coriaria Lactone could increase the average opening time of Ca2+-activated K+ channels (1.867±0.210, 6.900±0.120, P ＜ 0.01), and reducing the average closing time (78.505±7.192,6.233±0.854, P ＜ 0.01).CONCLUSION: In epilepsy induced by Coriaria Lactone, the activation of Ca2+-activated K+ channels might play an important role of negative modulation.

Objective:To investigate the effect of Rab7 on cytokine induced by TLR7 (Toll like receptor-7) R848 activated in Raw264.7,and discusses the influence of Rab7 on MAPK signal transduction.Methods: TLR7 downstream cytokines such as TNF-α,IL-6,IFN-α,IFN-β and IP-10 activated by R848 were detected through Q-PCR in Rab7 silenced mouse macrophages,and then analysis of phosphorylation of MAPK determined with Western blot showed the effect of Rab7 on signal transduction of MAPK.Results: Rab7 inhibit production of cytokine activated by TLR7,and also,Rab7 had an inhibitory effect on MAPK signal pathway.Conclusion: The experimental results further illustrate that the Rab7 is the TLR7 signal transduction pathway negative regulatory factor,and to participate in MAPK signaling pathway.