Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.

Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.

AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.

Child ; Chondrosarcoma, Mesenchymal ; diagnosis ; drug therapy ; pathology ; Diagnosis, Differential ; Humans ; Male ; Pelvic Neoplasms ; diagnosis ; drug therapy ; pathology ; Tomography, X-Ray Computed ; Treatment Outcome

Objective:To investigate the effect of the ceramometal b ridge on the local gingival groove flora.Methods: Classi cal bacterial incubation and identification were used to study the changes of th e local gingival groove flora in 6 patients wearing ceramometal bridge for 1 wee k to 3 months. Results: 3 months after the prosthetic procedure the CFU o f P.melaninogenica, Gram-positive bacilli cmainly Actinomyces and the t otal bacteria were significantly increased (P

Asteroid Hyalosis (AH) is a common clinical disease,which has been considered a benign disorder as it rarely impairs visual acuity.It was often discovered when the patient was treated for other eye diseases.The mechanism was unclear.Its characteristic B-ultrasound property makes the B-ultrasound a very helpful diagnostic technique.In the case of the patients with other fundus diseases associated with AH,optical coherence tomography (OCT) and fluorescein angiography (FA) may be used to reduce the interference from asteroid bodies,therefore improve the fundus visibility.Recent studies have shown that AH can incorporate with many other eye diseases.For example,in patients with cataracts,asteroid hyalosis can cause surface calcification of silicone plate intraocular lenses,which in most cases may lead to the need for explantation of the calcified intraocular lenses.The efficacy of pars plana vitrectomy (PPV),the removal of some,or all,of the eye`s vitreous humor for AH remains controversial.In this paper,we provide a review of the recent literature on AH disease: the etiology,diagnosis and treatment.We hope to thus improve the awareness and outcomes of AH disease.

AIM:To investigate the timing and efficacy of vitrectomy for patients with vitreous hemorrhage(VH) due to proliferative diabetic retinopathy(PDR).METHODS:Retrospective analysis.Patients who presented to our hospital between Feburary 2012 and May 2014 with VH secondary to PDR treated with vitrectomy were included.All patients were divided into three groups according to the duration of VH.A group was less than 1mo for 22 eyes, B group was 1-3mo for 23 eyes, C group was more than 3mo for 25 eyes.All patients underwent intravitreal injection of ranibizumab 1-2wk before vitrectomy, and supplemented or finished panretinal photocoagulation (PRP) intraoperatively or postoperatively.Patients with cataract accepted phacoemulsification and intraocular lens implantation.Eyes filling silicone oil were implanted intraocular lens in the second phase.All patients were followed up 24 to 42mo (mean:28.7mo).We assessed the intraoperative complications such as hemorrhage, iatrogenic retinal hole, and postoperative complications such as vitreous hemorrhage, neovascular glaucoma.Macular edema and best corrected visual acuity were observed at every follow-up.RESULTS:There was no significant difference for other baseline data (P>0.05) but DR stage between three groups (P=0.033).There was significant difference of last follow up visual acuity between three groups (P<0.001).The significant difference can be seen between group A and B (P=0.03).The same outcome showed between Group A and C(P<0.001).There was no significant difference between Group B and C (P>0.05).The percentage of visual acuity was 0.5 and above in the three groups were:41%, 23%, 0 respectively.The patients with visual acuity of less than 0.1 were 5%, 26% and 40% respectively.Silicone oil filling rate of three groups were:9%, 26%, 40% respectively and there was no significantly difference between three groups on postoperative complications (P>0.05).CONCLUSION:Patients with VH due to proliferative diabetic retinopathy undergoing early vitrectomy may get better visual acuity than who accepting delayed vitrectomy.

Using of clearing zones on pachyman agar medium, there are 217 plant endophytic actinomycetes, producing ?-1,3-glucanase were screened. 45.6% of the strains produced ?-1,3-glucanase, in which the strains from cucumber are up to 38. The percentages of endophytic actinomycetes from different hosts produceing ?-1,3-glucanases were different. The percentage of the strains in Rhizoma Polygonatum produced ?-1,3-glucanases is the highest, up to 88.9%. The Inhibited effects of plant endophytic actinomycetes which produced extracellular ?-1,3-glucanases on mycelium growth of Sclerotinia sclerotiorum were detected in vitro. Cucumber endophytic actinomycete gCLA4 strain was screened out from 99 isolates, which can strongly inhibit the growth of S. sclerotiorum. The optimal ?-1,3-glucanases fermentation conditions of strain gCLA4 were investigated, they were pachyman 0.2%, peptone as nitrogen, pH 7～8 for 5 days. The ?-1,3-glucanases of strain gCLA4 had some inhibiting efficiency on 13 plant pathogens, in which inhibiting efficiency to Botryosphaeria dothidea was the strongest.

0.05).The concentration of urinary KYNA,metabolite of the KYN,was significantly lower in SHRs compared to WKYs(7.8?1.8 vs 19.9?3.5 ?mol/24 h P=0.013).Both KAT activity in renal cortex and KYNA content in urine were negatively correlated to blood pressure(r=-0.418,P=0.023;r=-0.723,P=0.001).Conclusion The declined activity of KAT in renal cortex and the deficiency of KYNA concentration in urinary may affect blood pressure regulation in SHR by renal metabolite of the KYN.