OBJECTIVETo observe the changes in the adhesive function of vascular endothelial cells (VEC) and rat monocytes induced by lipopolysaccharide (LPS) and co-cultured with smooth muscle cells (SMC) and the intervention effect of paeonol (Pae).

METHODPrimary rat vascular endothelial cells (VECs) and rat vascular smooth muscle cells (VSMCs) were cultured by predigesting and adhering tissue blocks. The VEC-VSMC co-culture model was established by Transwell chamber. LPS was used to induce VEC injury. MTT assay and LDH assay were used to determine the VEC activity. ELISA assay was used to detect IL-1beta and TNF-alpha secreted by the VEC. The immunocytochemistry assay was carried out to detect the expression of ICAM-1. The Rose Bengal Staining was used to test adhesive function between VECs and monocytes.

RESULTThe concentration of LPS-induced VEC injury was 100 microg x L(-1), and the time was 7 h. after the intervention on the above cell model for 24 h, Paeonol (15, 30, 60 micromol x L(-1)) could effectively inhibit LPS-induced VEC injury and VEC injury, significantly enhance the survival rate of LPS-injured VECs, decrease IL-1beta and TNF-alpha secreted by the injured VEC, and reduce the expression of ICAM-1, so as to inhibit the adhesion of LPS-induced VECs and monocytes.

CONCLUSIONPaeonol could inhibit IL-1beta and TNF-alpha expression to protect VECs from being injured by LPS, and reduce ICAM-1 expression to inhibit the adhesion between VECs and monocytes.

Acetophenones ; pharmacology ; Animals ; Cell Adhesion ; drug effects ; Coculture Techniques ; Endothelial Cells ; cytology ; drug effects ; metabolism ; secretion ; Gene Expression Regulation ; drug effects ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; pharmacology ; Male ; Monocytes ; cytology ; drug effects ; Myocytes, Smooth Muscle ; cytology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVE To know the incidence, etiology, risk factors of hospital-acquired pneumonia(HAP). METHODS From Jan 2003 to Dec 2004, 129 cases of HAP were retrospectively studied. Epidemiologic data, etiology , and evolution of pneumonia were recorded. RESULTS The 115 patients with HAP had one or more intrinsic risk factors, mainly malnutrition (albumin values

[Objective] To explore the effects of angongniuhuang pill on the synthesis of apolipoprotein E in brains of rats after traumatic brain injury.[Methods] Using reverse transcription-polymerase chain reaction(RT-PCR) and enzyme linked immunosorbent assay(ELISA),the apolipoprotein E’s mRNA of contused brain tissue and the concentration of apolipoprotein E in CSF at different phases after traumatic brain injury were detected of rats fed with angongniuhuang pills.[Results]High expression of apolipoprotein E mRNA and high concentration of apolipoprotein E were found in rats fed with angongniuhuang pills compared with controls.[Conclusion]Angongniuhuang pills may increase the expression of apolipoprotein E mRNA in brains after traumatic brain injury,thus promote the synthesizing and secreting of apolipoprotein E.

AIM To establish an HPLC method for the simuhaneous content determination of five flavonoids in Duchesnea indica (Andr.) Focke at five picking time (April,May,June,July and August).METHODS The analysis of D.indica methanol extract was performed on a 25 ℃ thermostatic Agilent ZORBAX SB-C18 column (250 mm×4.6 mm,5 μm),the mobile phase comprising of acetointrile-0.1％ methanoic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 350 nm.RESULTS Rutin,hyperoside,isoquercitrin,celereoin and kaempferol showed good linear relationships within their own ranges (R2 ≥0.998 6),whose average recoveries were 97.1％-101.5％ with the RSDs of 1.37％-2.37％.The contents of five constituents in samples at different picking time exhibited obvious differences,among which lutin and hyperoside contents were the highest in June,isoquercitrin content was the highest in July,and celereoin and kaempferol contents were the highest in August.CONCLUSION The suitable picking time of D.indica is June and July.

Objective To investigate the long-term effects of xanthine oxidase inhibitor,oxypurinol on myocardial contractility of post-ischemic heart failure in mice,and explore the underlying mechanism. Methods One hundred and twenty SV120 mice were randomly assigned into myocardial infarction control group,sham operation group and Oxy treatment group.Post-ischemic heart failure were induced by left anterior descending coronary artery ligation in myocardial infarction control group and Oxy treatment group,and mice in Oxy treatment group and sham operation group were orally administered with 0.5 mmol/L Oxy each day.Nine to eleven months after treatment,echocardiography was performed in all groups.Trabeculae from the right ventricle of mice were dissected for assessment of changes in excitation-contraction coupling.Sarcomere length was measured by laser diffraction.Intracellular free Ca~(2+) concentration([Ca~(2+)]_I)was detected with fluorescent dye Fura-2,which was microinjected iontophoretically into cells. Steady-state force-[Ca~(2+)]_I was achieved by addition of ryanodine and increasing the stimulus frequency to induce tetanization,and the relationship between myocardial contractility and intracellular Ca~(2+) transients was analysed.Besides,Western blotting was performed to determine the oxidation of myofilament proteins. Results Long-term oral administration of oxypurinol significantly improved myocardial contraction function and reduced ventricular wall thickness.Programming of excitation-contraction coupling was significantly improved,and maximal Ca~(2+) activated force(F_(max))in steady-state wag also significantly increased.Western blotting revealed the oxidative modification of actin in mice of Oxy treatment group was significantly inhibited compared with that of myocardial infarction control group. Conclusion Long-term treatment with Oxy improves the cardiac contraction function and boosts the cardiac force dramatically in post-ischemia heart failure.The increase in contraction is the result of increased myofilament Ca~(2+) responsiveness.Thus,antioxidant oxypurinol,by preventing oxidative damage to contractile proteins,can augment contraction with little changes in[Ca~(2+)]_I,represents new class of inotropic agents with advantages of reducing Ca~(2+) overload,and offers new promises in management of heart failure in the future.

Objective To observe the expression of immune substances secreted by peripheral bloodγδT cells after stimulated by early secreted antigenic target-6(ESAT-6),and explore its mechanism of signaling pathway.Methods Peripheral blood mononucle-ar cells(PBMC)were collected from health human blood with the ficoll density gradient centrifugation method,and theγδT cells were separated from the PBMC with flow cytometry;the inhibitor of TLR-4 signaling pathway(E5564)was used to cocultured withγδT cells to inhibit the function of TLR-4,and the change of TLR-4 was analyzed by the methods of PCR and Western blot.ESAT-6 were used to stimulate theγδT cells,and the control group without any stimulating factor was established,then the expression levels of IL-17,TNF-α,IFN-γwere determined by ELISA method after 0、1、3、6、9 and 12 days.Results The results of PCR and Western blot showed that ESAT-6 could increase the expression of TLR-4(P<0.01);The results of ELISA showed that ESAT-6 could enhance the expression of IL-17,TNF-αand IFN-γ(P<0.01),and the inhibitor of TLR-4(E5564)could decrease the expres-sion of IL-17,INF-α,IFN-γ(P<0.01).Conclusion ESAT-6 can induceγδT cells to produce more IL-17,TNF-α,IFN-γ,and the mechanism of which maybe concerned with TLR-4 signaling pathway.

Objective To explore the effect of recombinant human tumor necrosis factor-α receptor Ⅱ:IgG Fc fusion protein injection (rhTNFR:FC) on the expression of cartilage oligomeric matrix protein (COMP) in the synovial fluid and peripheral blood of juvenile idiopathic arthritis (JIA); and to explore the clinical significance of COMP for JIA and the relationship between rhTNFR:FC and COMP in JIA.Methods Thirty-five patients with JIA (JIA group),30 patients with traumatic arthritis (trauma group) and 30 patients with indirect inguinal hernia hernioplasty (normal group) were included.Peripheral blood from all enrolled patients and synovial fluid from 15 JIA and 10 trauma arthritis were obtained for COMP detection before the treatment.Fifteen JIA (group A) patients were treated with combined rhTNFR:FC,diseasemodifying antirheumatic drugs (DMARDs) and non-steroid anti-inflammatory drugs (NSAIDs),20 JIA (group B) were treated with combined DMARDs and NSAIDs.After three to six months' treatment and when the disease were in remission,peripheral blood from group A and B were drawn for COMP detection.In group A,the synovial fluid from 5 patients were obtained for COMP detection after treatment.At the same time,such as tender joint count (TJC),swollen joint count (SJC),time for morning stiffness,blood routine,erythrocyte sedimentation rate (ESR),and C-reactive protein (CRP) and other parameters before and after treatment were measured.The level of COMP was tested by double antibody sandwich enzyme-linked immunosorbent assay.The measurement data were tested for variance and independent sample t-test; and the enumeration data were tested by chi-squared or Fisher's exact test.Pearson's correlation analysis was adopted to analyze the association among the variables.Results ① The blood COMP level before treatment was (0.77±0.29) ng/ml in the JIA group,(1.00±0.28) ng/ml in the traumatic arthritis group,and (1.33±0.37) ng/ml in the normal control group.The level in the former two groups was obviously lower than that in the normal control group.The variation was statistically significant (F=25.345,P＜0.05).The comparison between any two groups was statistically significant (P＜0.05).② The COMP level in the synovial fluid before treatment were (14.8±1.6) ng/ml in the JIA group,(15.1±1.0) ng/ml in the traumatic arthritis group.The variation was not stati-stically significant (t=0.523,P=0.606).③ The serum COMP level of the systemic JIA group was obviously lower than that of the oligoarticular JIA patients,and patients with enthesitis-related arthritis and polyarticular JIA (0.26± 0.03 vs.0.87±0.17,0.89±0.22 and 0.70±0.35 ng/ml,respectively; F=9.244,P＜0.05).④ The serum COMP level of JIA at the acute phase was negatively correlated with white blood cells count (WBC),CRP and ESR (r=-0.556,-0.582 and-0.684,respectively; P all＜0.05).By contrast,no correlation was detected between the serum COMP level and joint tenderness index,joint swelling index,morning stiffness duration,hemoglobin level and platelet count(r=0.06,-0.206,-0.107,0.15 and-0.185,respectively; P all ＞0.05).⑤ The serum COMP level was obviously lower in the JIA with joint destruction than that without joint destruction (0.52±0.22 vs.0.92±0.22 ng/ml; t=5.207,P＜0.05).⑥After treatment,the blood COMP level in group A was (1.33±0.21) ng/ml and (0.96±0.22) ng/ml in group B,which was obviously higher than that in the JIA group before treatment (0.77±0.29) ng/ml.In addition,the level in group A was higher than that in group B.The variation was statistically significant (F=24.681,P＜0.05).⑦ After treatment,the COMP level in the synovial fluid (18.4± 1.1) ng/ml (n=5) was higher than that before the treatment was (14.8± 1.6) ng/ml (n =15).The variation was of statistical significant (t=4.565,P＜0.05).Conclusion The COMP level in blood and synovial fluid declines before treatment and increases after treatment.The increase is more obvious after combined with rhTNFR:FC treatment.The serum COMP level is remarkably decreased in JIA at the acute phase,systemic JIA,and the JIA with destruction of joint,and showes a negative correlation with WBC,CRP and ESR.Serum COMP may be a useful marker of active disease,destruction of joint and growth inhibition for patients with JIA.rhTNFR:FC treatment for JIA can facilitate the recovery of COMP.

Objective To investigate the effect of carteolol hydrochloride eye drops on 24-hour variation of intraocular pressure (IOP). Methods The 30 patients who had never used any anti-glaucoma eye drops were chosen, and their 24-hour variation of IOP (at 2:00 AM, 6:00 AM, 10:00 AM, 2:00 PM, 6:00 PM, 10:00 PM) were measured. Then carteolol hydrochloride eye drops were used twice a day (7:00 AM and 7:00 PM), 1-2 drops each time. Fifteen days later, their 24-h IOP was measured again and the data were analyzed. Results The average IOP at six monitoring points before treatment were (19.77±2.51) mm Hg, (19.52±2.34) mm Hg, (20.38±2.71) mm Hg, (20.07±3.65) mm Hg, (19.97±4.10) mm Hg and (19.97±4.31) mm Hg, the corresponding data after 15 days of treatment were (17.12±2.38) mm Hg, (17.42±2.60) mm Hg, (18.18±2.36) mm Hg, (18.02±3.77) mm Hg, (17.72±3.17) mm Hg and (17.35±3.40) mm Hg (t=11.68, P=0.000). The difference before and after treatment was (2.46±2.05) mm Hg during day and (2.17±1.88) mm Hg during night, and no significant difference was found between them. Conclusions Carteolol hydrochloride eye drops could significantly reduce the IOP, the decreased extent of 24-h IOP is stable.

Objective To summarize the preliminary experience of coils, Onyx and covered stents for the treatment of carotid-cavernous sinus fistula (CCF). Methods Of the 25patients with CCF, 22 were traumatic CCF, and 3 were spontaneous CCF; the fistula orifices of 14 patients were located in the cavernous segment of right internal carotid artery (ICA), and 11were located in the cavernous segment of left ICA. According to the Barrow classification,Types A, C, and D were 22,2, and 1, respectively. Eighteen patients were treated with Guglielmi detachable coils alone, 1 was treated with Guglielmi detachable coils and Onyx, 2 were treated with Onyx alone, and 4 were treated with covered stents. Results Intracranial bruit disappeared immediately in all patients after the procedures. Postoperative angiography showed complete occlusion of the fistula orifices, the symptoms disappeared after 1 to 20 days, and there were no death and procedure-related complications. The recanalized ICAs were patent in patients (12/18) treated with coils, and the ICAs were patent in 4 patients treated with the covered stents.The ICAs were patent in 2 patients treated with Onyx, the ICA was patent in 1 patent treated with detachable coils and Onyx. One patient with traumatic CCF treated with coils had clinical recurrence, and was cured after re-embolization with coils. The remaining patients were followed-up for 6 to 24 months, and no recurrence of clinical symptoms was found. The followup cerebral angiography of 6 patients was obtained, and no CCF recurrence was observed.Conclusion Endovascular treatment with Guglielmi detachable coils, Onyx, and covered stents is a safe and useful method for closing CCFs.

Objective To explore the value of combining Uniplate anterior cervical plate system with titanium cage to treat single level cervical spondylotic myelopathy. Method Observing the clinical outcome and X-ray results of 32 patients with cervical spondylotic myelopathy undergone anterior decompression, autograft fusion and internal fixation with Uniplate anterior cervical plate system and titanium cage. Results All patients were followed up for 3 to 12 months. According to bony fusion criteria,no implants breakage, dislocation or loosening was found in all cases in 3 months' postoperatively radiological review. According to Odom classification,excellent outcome was 27 patients, good was 5 patients, the excellent and good rate was 100%(32/32). Conclusions Combining Uniplate anterior cervical plate system with titanium cage to treat single level cervical spondylotic myelopathy has advantages of easy manipulation,safe, less complication and higher fusion rate. It is an ideal way for cervical anterior fixation.