Objective:To detect and compare the intensity of gingipain K(Kgp)in culture medium and cell extract of Porphyromonas gingivalis(P.gingivalis)isolates in puberty gingivitis,and then to reveal the possible relationship between Kgp and puberty gingivitis.Methods:36 patients with puberty gingivitis aged from 14 to 17 years were enrolled.Clinical parameters including GI,SBI and PD were evaluated before subgingival plaque samples collection.Subgingival plaque samples were collected and then P.gingivalis isolates were obtained.16S rRNA PCR was used to confirm the presence of P.gingivalis in clinical isolates.Bacteria were cultured in BHI agar base and harvested at the end of log-phase growth.Culture fractions of P.gingivalis(culture medium and cell extracts)were performed with SDS-PAGE and Western blot technique using primary antibody against specific anti-Kgp N-terminal IgG subdomain.The data were statistically analyzed using SPSS 11.5 software.The relationship between the Kgp intensity and the clinical parameters was statistically analyzed using sum rank test.Results:There was positive correlation between the intensity of Kgp N-terminal IgG subdomain and the clinical parameters(P

Objective To investigate the effect of ghrelin on PAI-1 secretion in HepG2 cells induced by TNF-αand the effect of p-38 MAPK.Methods HepG2 cells were cultured.The concentration of TNF-α used to treat the HepG2 cells wag selected.The effect of ghrelin on PAI-1 secretion induced by TNF-α was detected by ELISA,the p-38 MAPK expression was investigated by Western blot.Results The concentration of PAI-1 was increased when cells were exposed to different concentration of TNF-α.The p-p38 MAPK expression was increased when the cells were exposed to TNF-α,ghrelin could inhibit the increase of PAI-1 secretioN induced by TNF-α.The expression of p-p38 MAPK was decreased when the cells were pretreated with ghrelin.Conclusion PAI-1 secretion were increased after TNF-α in-creasing.Ghrelin could inhibit PAI-1 secretion via p38 MAPK.

This study was aimed to observe four kinds of kidney-tonification medicine, which were Epimedium, pso-ralen, Ligustrum lucidum, Polygonum with the active ingredient of icariin, psoralen, oleanolic acid, stilbene glucoside and their orthogonal compatibility. There were two kinds of non-kidney tonification medicine, which were Chuanx-iong and astragalus with the active ingredient of TMP and astragaloside. The observation was made on the regulatory role of rat bone marrow stem cells (BMSCs). A total of 65 SD rats were randomly divided into the normal control group, positive transformed control group, kidney-tonification compatibility group (including Group 1, Group 2, Group 3, Group 4, Group 5, Group 6, Group 7, Group 8, and Group 9), non-kidney tonification medicine control group (in-cluding TMP group and astragaloside group). Intragastric administration of medication was given to the kidney-tonifi-cation compatibility group and the non-kidney tonification medicine control group, once a day for 3 consecutive days. Intragastric administration of equal amount of normal saline was given to the normal control group and the posi-tive transformed control group. On the third day of intragastric administration, rats in each group were sacrificed. Serum containing medication was used in the culture of BMSCs for 6, 12, or 18 days. ELISA method was used to quantitatively detect the expression activity and content of BMP2 on the 6th, 12th, or 18th day, in order to evaluate the degree of bone cell differentiation degree. Real-time quantitative PCR method was used for detection of expression of Bmp2, Smad1, Smad4 mRNA in serum containing medication in the culture of BMSCs on the 18th day. The results showed that the kidney-tonification compatibility can improve the expression activity and content of BMP2 culture in vitro, with the peak on the 12th day. The kidney-tonification compatibility groups can upregulate expressions of Bmp2, Smad1, Smad4 mRNA. It was concluded that the active ingredient compatibility of kidney-tonification medicine can promote BMSCs. Its mechanism may be related to the upregulation of expression of Bmp2, Smad1, Smad4 mRNA, and the activity and content of Bmp2.

98% amino acid identity.There are 28 different amino acids between them,with 7 of which locating in the region encoded by exon6A or exon6.Alternative splicing of exon18 was not found in the gene cloning of human brain Nav1.5/SCN5A,which was different from human heart Nav1.5/SCN5A,but a novel alternative splicing lacking exon24 was first found.The two variants were detected in similar ratio in brain,but they were proved to relate to age development in heart tissue.The exon24 of human Nav1.5/SCN5A has 54 nucleotides,encoding 30 amino acid residues,and are located in human chromosome 3P21.This alternative splicing was also found in other tissues other than heart and brain.The expression pattern of the two variants in different tissues was different when detected by competitive PCR method and it was also changing with age development.Furthermore,Nav1.5/SCN5A mRNA was detected in 16 different tissue types of Wistar rats(P80) by reverse polymerase chain reaction(RT-PCR) .These results suggest that Nav1.5 Na+ channels in human brain are encoded by new variants of Nav1.5/SCN5A and its mRAN is more widely expressed than previously thought.The study is useful for making further investigation in the functional analysis of Nav1.5 Na+ channels in different tissues.

ObjectiveTo investigate the expression of TGF ?1 and type Ⅰ receptor and their relations with intimal hyperplasia in autogenous vein grafts in rats. Methods Autogenous vein graft model was established in 48 Wistar rats. The vein grafts were harvested on day 3, 7, 14, and 28.Histomorphological methods were used to measure the thickness of intima and wall at different time points. Immunohistochemistry and Western blot were used to detect the protein expression of TGF ?1 and TGF ?RⅠ . RT PCR was used to detect their mRNA level. ResultsThe intimal thickness increased on day 7 compared with controls( P

AIM To investigate whether Aβ deposit in Alzheimer disease(AD) impairs signal transduction pathway responsible for neuronal survival.METHODSThe rats were randomly divided into six groups:control group and Aβ25-35 group,Aβ25-35+ibuprofen groups (7.5 and 15 mg·kg-1,respectively),Aβ25-35+ibuprofen+LY294002 group,and Aβ25-35+LY294002 group.Rats were given ibuprofen (7.5 and 15 mg·kg-1 daily,ig) for 3 weeks prior to and 1 week after icv single dose of Aβ25-35 (10 μL,1 mmol·L-1).LY294002 was injected icv 1 h before the injection of Aβ25-35.Seven days after Aβ25-35 injection,the hippocampal expressions of P53,Bax,Fas ligand (FasL),Bcl-2 proteins,phospho-Akt/PKB,and phosphorylated 70 ku ribosomal protein S6 kinase (p70S6K) and caspase 3 were determined in the brain tissue preparations from CA1 area with Western blot.The activity of caspase 3 was measured using a caspase 3 colorimetric activity assay kit.RT-PCR was used to show the change of p70s6k mRNA level.RESULTS Aβ25-35 icv injection significantly down-regulated phosphorylated Akt/PKB from 1.32±0.14 to 0.69±0.08 and p70S6K from 0.769±0.028 to 0.479±0.032 in hippocampal CA1 region.These changes were accompanied by increased expressions of the proapoptotic proteins P53,Bax,and FasL and decreased expression of the anti-apoptotic protein Bcl-2 in rat hippocampus.In addition,caspase 3 activity was significantly enhanced in hippocampal CA1 region in Aβ25-35-treated rats compared with control rats.Ibuprofen can reverse these Aβ25-35-induced changes.CONCLUSION Down-regulated anti-apoptotic PI3K/Akt/p70S6K signaling pathway induced by Aβ25-35 in rat hippocampus may contribute to the neuronal damage in AD.Ibuprofen prevents Aβ25-35-induced down-regulation of PI3K/Akt/p70S6K signaling pathway.

AIM To observe the neuroprotective effect and protective mechanisms of ibuprofen on amyloid β-protein fragment 1-40 (Aβ1-40)-induced neurotoxicity in rat hippocampus.METHODS Rats were given ibuprofen (15 mg·kg-1 daily,ig) for 3 weeks prior to icv single dose of Aβ1-40 (5 μL,1 mmol·L-1).Six hours after Aβ1-40 injection,Western blotting was used to determine the expressions of phospho-MAP kinase kinase (MKK)3/MKK6,phospho-p38 MAP kinase,phospho-MAP kinase activating protein kinase 2 (MAPKAPK2),heat-shock protein 27(Hsp27),procaspase 9,3,and 7 cleavage,and poly (ADP-ribose) polymerase (PARP) cleavage in hippocampal CA1 region.RESULTS Intracerebroventricular injection of Aβ1-40 induced an increase in phosphorylated MKK3/MKK6 and p38 MAP kinase expressions in hippocampal CA1.These increases,in combination with reduced phospho-MAPKAPK2 and phospho-Hsp27 expressions,mediated Aβ1-40-induced the activation of caspases cascades.Ibuprofen (15 mg·kg-1·d-1,3 weeks) significantly prevented Aβ1-40-induced increases in phosphorylated MKK3/MKK6 and p38 MAP kinase expressions.In addition,Aβ1-40-induced decreases in phosphorylated MAPKAPK2 and Hsp27 expressions were abrogated by ibuprofen.Aβ1-40-induced changes in activation of caspases cascades were inhibited by ibuprofen.CONCLUSIONIbuprofen prevents Aβ1-40-induced neurotoxicity through suppression of phosphorylated MKK3/MKK6 and p38 MAP kinase expressions and the up-regulation of phospho-Hsp27 expression.

AIM To explore the mechanism of amyloid β-protein fragment 25-35(Aβ25-35)-induced inflammation and apoptosis in rat hippocampus in vivo by studying mitogen-activated protein kinase (MAPK) signaling pathway and the protective effect of anti-inflammatory drug ibuprofen. METHODS Rats were given ibuprofen (7.5 mg·kg-1 daily, ig) for 3 weeks prior to and 1 week after icv single dose of Aβ25-35 (10 μL, 1 mmol·L-1). Seven days after injection, Nissl staining and immunocytochemical technique were employed to determine the morphology of pyramidal neurons and astrocyte infiltration in hippocampal CA1. The expressions of IL-1β, extracellular signal-regulated kinase (ERK), p38 MAPK, PKC, and caspase-3 were determined by Western blot. Reverse transcription-PCR analysis showed changes in IL-1β mRNA level. RESULTS Intracerebroventricular injection of Aβ25-35 elicited astrocyte activation and infiltration and caused a strong inflammatory reaction characterized by increased IL-1β production and elevated IL-1β mRNA level. The inflammatory reaction was accompanied by the loss of pyramidal neurons in hippocampal CA1. The phosphorylation of p38 MAPK was significantly increased, on the other hand, the phosphorylation of ERK was significantly reduced and these were coupled with the increase of caspase-3 expression in hippocampal CA1. Ibuprofen (7.5 mg·kg-1 daily, 4 weeks) significantly reduced Aβ-induced IL-1β expression, caspase-3 expression and p38 MAPK activation. The loss of pyramidal neurons was also significantly attenuated by treatment with ibuprofen. CONCLUSION The activation of p38 MAPK and the down-regulation of ERK play a pivotal role in the inflam-matory response and apoptosis evoked by Aβ25-35 in vivo, which can be prevented by ibuprofen.

Objective To test the levels of vascular endothelial growth factor (VEGF) in bronchoalveolar lavage fluid (BALF) during acute lung injury (ALI)/acute respiratory distress syndome (ARDS) after the interference with anticoagulation and fibrinolytic in an attempt to find a possible treatment plan for ALI/ARDS.Methods Oleic acid-induced ALI models were used in this study.A total of 96 rats were randomly assigned oleic acid group,salvianolate group,urokinase group,and saline group according to the random number table,with 24 animals per group.In oleic acid group,oleic acid (0.2 ml/kg,caudally) was administered alone.In addition to the oleic acid,the rats in salvianolate and urokinase groups received caudal injection with 10 ml/kg of 0.06％ salvianolate and 5,000 U/kg of urokinase respectively.In saline group,saline solution with an equal volume of oleic acid was caudally administered alone.Effect of the early use of salvianolate and urokinase on levels of VEGF in BALF was measured.Clinically,patients with ALI/ARDS composing survivor and death groups were enrolled and assessed by acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and multiple organ dysfunction syndrome (MODS) score.Besides,healthy population was included as the controls to comparatively analyze the levels of VEGF in BALF in ALI/ARDS group.Results Of the rat models of ALI,early use of salvianolate or urokinase was associated with elevated level of VEGF in BALF (P ＜0.05),which subsequently attenuated the lung injury.In the clinical research,it was found that the level of VEGF in BALF in ALI/ARDS group was much lower than that in control group [(45.9 ± 5.7) pg/ml vs (60.6 ± 4.5) pg/ml] (P ＜ 0.01).The level of VEGF in BALF was negatively correlated with APACHE Ⅱor MODS scores (r =-0.542,-0.659,P ＜ 0.01).Moreover,patients in death group appeared lower VEGF level in BALF than those in survivor group(t =2.68,P ＜ 0.05).Conclusion VEGF level in BALF can be taken as an indicator of patient condition and outcome.Early use of anticoagulation or thrombolytic drugs may play a positive role on patients with ALI/ARDS.

Objective Aimed to clarify the molecular mechanism after subarachnoid hemorrhage （SAH） by investigating the expression of tight junction protein Claudin-5 and ZO-1 and the effects of SP600125 on them. Methods Seventy-five male Sprague Dawley rats （300 to 350 g） were randomly divided into sham,SAH,SAH ＋ DMSO （dimethyl sufoxide） solution,SAH ＋SP600125 （C-Jun N-terminal kinase inhibitor）10 mg/kg,and SAH ＋SP600125 30 mg/kg groups. The standard endovaseular perforation was performed to produce experimental SAH. The JNK inhibitor SP600125 was intraperitoneally administered at 1 hour before and 6 hours after SAH. Results At 24 hours after SAH,signs of microvessels injury were observed in brain cortex. Compared with the sham group,expression of Claudin-5 and ZO-1 was sig- nificantly decreased （P 〈 0.05 ）. JNK inhibitior SP600125 suppressed the decrease of Claudin-5 and ZO-1 expression, attenuated blood-brain barrier disruption in rats after SAH. Conclusions The blood-brain barrier disruption is an important mechanism of early brain injury after SAH. JNK inhibitor SP600125 improves neurological outcomes and provides neuropmtecfion against acute events after SAH such as bloodbrain barrier disruption and cell apoptosis.