BACKGROUND:Previous clinical fol ow-up study showed that disc degeneration of adjacent segment after anterior cervical discectomy and fusion was faster than that of artificial cervical disc replacement. Compared with the anterior cervical discectomy and fusion, artificial cervical disc replacement can maintain a good range of motion of replacement segment. Further investigation should be taken to compare the difference between stress and fusion after replacement. OBJECTIVE:To compare the adjacent level discs loads between artificial cervical disc replacement and anterior cervical discectomy and fusion. METHODS:A healthy 30-year-old male volunteer was scanned with CT at the artificial cervical intervertebral disc and anterior cervical plate. Three-dimensional images were reconstructed with Mimics 10.01 and Geomagic Studio.v11 software. Above three-dimensional data were input into the Abaqus6.9 finite element analysis software for meshing, assignment, and stress analysis. Finite element method was used to simulate the stress changes of the adjacent segments after artificial cervical disc replacement and anterior cervical discectomy and fusion. RESULTS AND CONCLUSION:(1) Under the same preload, during anteflexion, posterior extension, and lateroflexion, the disc stress at adjacent segment was significantly larger after anterior cervical discectomy and fusion than normal disc. Compared with normal persons, no significant difference was detected in stress of adjacent segment at anteflexion, posterior extension, and lateroflexion after artificial cervical disc replacement. (2) Compared with artificial cervical disc replacement group, the stress of adjacent segment increased 10.3%-51.6%in the anterior cervical discectomy and fusion group. (3) Finite element analysis showed that the stress was larger in the anterior cervical discectomy and fusion group than in the artificial cervical disc replacement group. With prolonged fol ow-up, compared with the conventional anterior decompression and fusion, artificial cervical disc replacement can better play its protective effect on the adjacent intervertebral disc.

AIM To study the effect of polysaccharides 2b from Mudan cortex (PSM 2b) on type 2 diabetes mellitus (T2DM) rats and explore its mechanism. METHODS T2DM rat model was established by low dosage streptozotocin and high sucrose fat diet. The drugs were administrated by ig for 5 weeks. The serum glucose was assayed with GOD POD method. Insulin was determined by radioimmunoassay. Insulin receptors of the plasma membrane from rat liver were determined with radioligand binding assay of receptors (RBAR). RESULTS PSM 2b could significantly lower fasting blood glucose (FBG), improve impaired glucose telarance (IGT) and dyslipidemia. It could also remarkably raise receptor total (RT2) of the low affinity insulin receptor (InsR) and insulin sensitivity index (ISI) in T2DM rats. CONCLUSION PSM 2b has an obvious therapeatic effect on T2DM in rats. Its mechamsm is relatively improve insulin resistance (IR) in T2DM rats by raising the number of InsR.

Objective To develop a gut-brain interaction animal model of IBS which combines multiple factors including behavior, visceral sensation and motility. Methods Setting up a multifactor interactional animal model (chronic acute combining stress model, CACS) based on a chronic unpredictable mild stress model of depression (CUMS) while combined with wrap restraint stress (WRS) , changes of some indexes were recorded including motility (granules of defecating, time of defecating), visceral sensitivity ( spontaneous contraction of abdominal striated muscles ) and behavior/mind ( sucrose consumption, body weight). G protein subunits were measured by Western blot in both hippocampus and prefrontal cortex simultaneously. Results ( 1 ) Compared with the state before stress given, defecating granules increased, defecating time of glassie from rectum shorten, number of abdominal contraction increased, and sucrose consumption decreased in CACS, however, neither significant change was found on defecating behavior in CUMS nor on sucrose consumption in WRS; (2) Compared with the control group, some G protein submits expression decreased in both CACS and CUMS( P < 0. 05 ) , while no significant changes of any G protein subunits were found in WRS. Conclusion The CACS animal model was a new, brain-gut interaction model, which can mimic part of human symptoms of IBS very well.

AIM:To investigate the effect of capsaicin on lipopolysaccharide ( LPS)-induced activation of cul-tured endothelial cells of mouse aorta in vitro.METHODS:The endothelial cells were isolated from mouse aorta and cul-tured in vitro, and the specific cell markers of the cells were identified by immunofluorescence staining.The cells were stimulated with LPS (100μg/L) combined with or without capsaicin, and the cells and supernatant were collected at 12 h, 24 h and 48 h.The levels of soluble intercellular adhesion molecule 1 ( sICAM-1) , soluble vascular cell adhesion molecule 1 (sVCAM-1) and soluble P-selectin (sP-selectin) in the supernatant were measured by ELISA.The levels of nuclear NF-κB p65 and cytopasmic p-IκBαand IκBαwere detected by Western blotting.RESULTS: Compared with control group, the levels of sP-selectin, sICAM-1 and sVCAM-1 in LPS group were significantly increased (P<0.05), and LPS promoted the expression of sICAM-1 and sVCAM-1 in a time-dependent manner.Compared with LPS group at the same time point, capsaicin inhibited the expression of sP-selectin, sICAM-1 and sVCAM-1 in a dose-dependent manner.Compared with con-trol group, the protein levels of NF-κB p65 and p-IκBαin LPS group at 24 h were significantly increased (P<0.05), while the protein level of IκBαin LPS group at 24 h were significantly decreased (P<0.05).Compared with LPS group, capsaicin decreased the protein levels of NF-κB p65 and p-IκBαand increased the protein level of IκBαin a dose-depend-ent manner.CONCLUSION:Capsaicin has a protective effect on LPS-induced vascular endothelial cell activation, which potentially contributes to the suppression of IκBαdegradation and NF-κB p65 nuclear translocation.

Objective To observe the release of glutamate (Glu)and γ-amino butyric acid (GABA) from PC 12 cells induced by aconitine,and to study the intervention of Shuanghuanglian on the injury of these cells. Methods The cell proliferation test agent in cell counting kit(CCK-8)was applied to assay the aconitine toxicity to PC12 cells and to establish the PC12 cell injury model induced by aconitine. The PC12 cells during logarithmic growing phase were randomly divided into the following groups:blank control group(complete medium containing 0.1% dimethyl sulfoxide was added), Shuanghuanglian control group (complete medium containing 50 μg/mL Shuanghuanglian),baicalin control group(complete medium containing 20 μmol/L baicalin),aconitine toxic group(complete medium containing 100 μmol/L aconitine),Shuanghuanglian intervention group(complete medium containing 100μmol/L aconitine and 50μg/mL Shuanghuanglian)and baicalin intervention group(complete medium containing 100 μmol/L aconitine and 20 μmol/L baicalin). The cells in all groups were incubated for 24 hours respectively. The changes of PC12 cell absorbance(A)values were detected by CCK-8 assay before and after intervention by Shuanghuanglian and baicalin. The PC12 cell apoptosis was determined by flow cytometry. Glu and GABA contents in cell culture medium were determined by chromatometry and enzyme-linked immunosorbent assay (ELISA). Results Compared with blank control group,after the PC12 cells treated with 100 μmol/L aconitine for 24 hours,their cytoactivity was decreased markedly(A value:1.003±0.042 vs. 1.685±0.030,P<0.05),then afterwards in the experiment,the incubation of 100 μmol/L aconitine with PC12 cells for 24 hours was considered as the intervention concentration. In blank control group,the normal PC12 cells accounted for 95.89%,while in the aconitine toxic group,the rate of injured PC12 cells reached 64.27% and early apoptosis rate reached 45.46%, and in Shuanghuanglian intervention group and baicalin intervention group,the early apoptosis rate was decreased to 33.24% and 28.22% respectively. Compared with blank control group,there were no significant differences in cytoactivities and the contents of Glu and GABA released by PC12 cells in Shuanghuanglian control group and baicalin control group(all P<0.05),while in the aconitine toxic group,the cytoactivity was significantly decreased(A value:1.056±0.039 vs. 1.722±0.083),and the contents of Glu and GABA were significantly increased〔Glu(μmol/L):5.295±0.137 vs. 3.433±0.138;GABA(μmol/L):0.769±0.020 vs. 0.528±0.012,both P<0.05〕. Compared with aconitine toxic group,the cytoactivities of PC12 were significantly elevated(1.202±0.059 and 1.180±0.032),the levels of Glu were significantly reduced(4.055±0.086 and 3.984±0.057),and the contents of GABA were obviously increased(0.809±0.016 and 0.930±0.021)in the cell culture medium of the Shuanghuanglian intervention group and baicalin intervention group(all P<0.05). The increase of cytoactivity in Shuanghuanglian intervention group was more marked than that of baicalin intervention group(P<0.05). There were no statistical significant differences in contents of Glu and GABA between Shuanghuanglian intervention group and baicalin intervention group(both P>0.05). Conclusions The changes of Glu and GABA may be one of the mechanisms of neural toxic effect of aconitine. Shuanghuanglian possibly can decrease Glu level and increase GABA content by way of its main component baicalin to antagonize the aconitine neurotoxicity.

OBJECTIVE To study the changes in matrix metalloproteinase-9 (MMP-9) mRNA, tissue-inhibitor of metalloproteinase-1 (TIMP-1) mRNA and the ratio of MMP-9 mRNA/TIMP-1 mRNA in the lungs of paraquate poisoning rats, and to investigate the protective effects of sodium dimercaptopropane sulfonate (DMPS, Unithiol). METHODS One hundred and twenty male SD rats were divided into 12 groups (n=10) randomly: normal control, DMPS control, PQ poisoning 1, 3, 7, 14 and 28 d model groups, (DMPS+PQ) 1, 3, 7, 14 and 28 d groups. PQ poisoning model was established by intraperitoneal injecting paraquate. The expression of MMP-9 mRNA and TIMP-1 mRNA in lung tissues was detected by RT-PCR. RESULTS ①By observing the changes of action and observing the lung tissues sections, the rats' PQ poisoning models was successfully established. ②The histopathology of lung showed infiltration of inflammatory cell in acute phase(within 2 weeks), the inflammation decreased gradually after 2 weeks, hyperplasia of collagen and pulmonary fibrosis were instead. Howerer, the pathological changes were alleviated obviously in the (DMPS+PQ) groups. ③Compared with the PQ groups, the expression of MMP-9 mRNA and TIMP-1 mRNA in lungs diminished greatly in the DMPS+PQ groups after rats injected DMPS(P<0.05); the ratio of MMP-9/TIMP-1 mRNA was bigger at 7, 14 and 28 d in the DMPS+PQ groups after rats injected DMPS. CONCLUSION Down-regulation of the expression of MMP-9 mRNA and TIMP-1 mRNA and up-regulation ratio of MMP-9 mRNA/TIMP-1 mRNA by DMPS may be one of mechanisms by which pulmonary injury and pulmonary fibrosis are prevented in the acute paraquate poisoning.

Objective To investigate clinical features,treatments and prognostic factors of the patients with necrotizing fasciitis caused by vibrio infections and thus provide reference for the early treatment and prognostic assessment.Methods A retrospective analysis was conducted on clinical data of 56 patients with vibrio necrotizing fasciitis admitted to the emergency center of our hospital from May 1995 to June 2011.The clinical characteristics and treatments of the patients were summarized,and differences of clinical factors between the survival group and death group were compared.The possible influencing factors for prognosis were also analyzed.Results The main clinical manifestations included fever (61％),shock (84％) and organ dysfunction,of which renal insufficiency (88％) was the most common,with case fatality of 43％.Early pathological changes of limbs were only local swelling and pain,while skin ecchymosis,tension blood blisters,necrosis and subcutaneous crepitation were the signs of advanced stage.Comprehensive treatment regime including early administration of sensitive antibiotics plus surgical incision and drainage and medicine support was given.A series of factors were significantly different between the survival and death groups including the duration from the presentation of symptoms to hospital admission (P ＜ 0.05 ),limb lesions involving the trunk (P ＜ 0.01 ),creatine kinase level (P ＜ 0.05 ),and emergency incision and drainage ( P ＜ 0.01 ).Conclusions The most prominent clinical manifestations of vibrio necrotizing fasciitis are rapidly progressive local symptoms and signs,and sharp deterioration of systemic conditions.Delayed visiting,severe local lesions,and failure to emergency surgery may be the factors for poor prognosis.

Objective To evaluate the relationship between changes in B-type natriuretic peptide(BNP) and cardiac troponin I(cTnI)levels and prognosis of critically ill patients with sepsis. Methods This study retrospectively reviewed the clinical data of 75 patients with severe sepsis and septic shock admitted into Emergency Intensive Care Unit(EICU)of the First Affiliated Hospital of Wenzhou Medical University in Zhejiang Province. According to the severity of the cases,they were divided into two groups:severe sepsis group(34 cases)and septic shock group(41 cases),and based on the difference in prognosis,they were divide into survivor group(32 cases) and non-survivor group(43 cases). Electrocardiogram(ECG)was performed within 24 hours after admission in all the patients. Acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score and biochemical markers showing organ dysfunctions as BNP, cTnI, creatine kinase (CK), creatine kinase MB mass(CK-MB), and lactate were compared between severe sepsis and septic shock groups and between survivor and non-survivor groups. Results The septic shock group had significantly higher baseline BNP,cTnI,lactate and APACHE Ⅱscore and mortality rate than those in severe sepsis group〔BNP(μg/L):1.90(1.08,2.79)vs. 0.41(0.31,0.75),cTnI (μg/L):1.15(0.92,1.28)vs. 0.58(0.40,0.79),lactate(mmol/L):6.63±3.72 vs. 3.28±1.66,APACHEⅡscore:26.00(24.00,28.00)vs. 21.50(20.00,29.25),mortality rate:70.73%vs. 41.18%,P<0.05 or P<0.01〕. Compared with survivor group,the ages of non-survivor group were older with more males and higher BNP,cTnI,lactate and APACHEⅡscore〔males(cases):30 vs. 13,age(years old):66.49±14.97 vs. 58.19±17.05,BNP:1.60(0.62, 2.51)vs. 0.57(0.37,1.79),lactate:4.10(3.00,9.00)vs. 3.10(2.13,4.18),cTnI:1.02±0.49 vs. 0.62±0.37, APACHE Ⅱ score:28.00(25.00,30.00)vs. 21.00(20.00,25.75),P<0.05 or P<0.01〕. However,there were no statistically significant differences in the levels of CK and CK-MB between the above compared groups(both P>0.05). The patients' ECGs had no obvious changes. Conclusions High plasma BNP and cTnI levels in patients with sepsis may suggest myocardial damage and relatively bad prognosis. The examination of BNP and cTnI levels may help clinicians to early detect the high-risk patients with septic cardiac dysfunction and assess their prognoses.

Objective To investigate the effect of resveratrol (Res) on paraquat (PQ)-induced acute lung injury (ALI) and mortality in mice and the mechanism of nuclear factor-κB (NF-κB) inflammatory pathway. Methods Sixty-eight healthy male ICR mice with grade SPF were enrolled, among them 20 mice were used for mortality observation (n = 10), and other 48 were used for determination of related parameters (n = 6). The mice were randomly divided into four group s: normal saline (NS) control group, Res control group, PQ group and PQ + Res group. The mice in the latter two groups were subdivided into 6, 24, 72 hours subgroups. The PQ poisoning model of mice was reproduced by one injection of 30 mg/kg PQ intraperitoneally. The mice in PQ + Res group were given 60 mg/kg Res intraperitoneally on the contralateral side after PQ injection. The mice were sacrificed at 6, 24, 72 hours after PQ poisoning, and lung tissue was harvested. The serum levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6 and IL-1β) were determined by enzyme linked immunosorbent assay (ELISA). The pathological changes in lung tissue were observed with electron microscopy. Apoptosis cells in the lung were identified by terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) for the estimation of apoptosis rate. The protein expression of NF-κB p65 was determined by Western Blot. Results Compared with PQ group, the death number of mice at 48, 72, 96 hours in PQ + Res group was slightly decreased (0 vs. 2, 2 vs. 5, 4 vs. 6) but without statistically significant difference (all P > 0.05). Under electron microscope, the lung injury in PQ group was severer than that in NS control group, and Res was found to be able to alleviate the lung injury. Compared with NS control group [(2.45±0.61)%], the apoptosis rate at 6 hours in PQ group was significantly increased [(8.42±1.48)%], and peaked at 72 hours [(21.23±3.47)%]. Res could decrease the apoptosis rate after PQ poisoning [6 hours: (5.56±1.31)% vs. (8.42±1.48)%, 24 hours: (11.14±2.07)% vs. (16.88±2.96)%, 72 hours: (13.28±2.32)% vs. (21.23±3.47)%, all P < 0.05]. The serum levels of TNF-α, IL-6, and IL-1β, and NF-κB p65 in lung tissue were all markedly increased after PQ poisoning, and they were significantly decreased after Res intervention as compared with those of PQ group [TNF-α (ng/L): 2.62±0.29 vs. 4.06±0.74 at 6 hours, 3.98±0.41 vs. 6.79±0.80 at 24 hours, 5.06±0.75 vs. 11.00±0.75 at 72 hours; IL-6 (ng/L): 14.19±1.54 vs. 16.55±1.24 at 6 hours, 13.21±1.37 vs. 19.73±0.85 at 24 hours, 13.72±0.56 vs. 22.45±0.72 at 72 hours; IL-1β (ng/L): 8.54±1.64 vs. 12.59±0.66 at 6 hours, 10.15±0.29 vs. 16.24±1.03 at 24 hours, 16.14±0.70 vs. 19.55±0.56 at 72 hours; 6-hour NF-κB p65: (1.34±0.07) folds vs. (1.86±0.11) folds when the expression in NS control group was represented as 1, all P < 0.05]. Conclusions Res cannot lower the mortality in mice with PQ poisoning, but it seems to be able to attenuate PQ-induced ALI and cell apoptosis. The mechanism responsible for the latter maybe the inhibitive effect of Res on NF-κB p65 translocation and cytokines production.

Objective To observe the dynamic changes of high mobility group protein B1 ( HMGB1 )expression in the lung of rats with Vibrio vulnificus sepsis so as to unravel the role of HMGB1 in lung injury.Methods Sixty rats of clean grade were randomly divided into normal control group ( A group, n = 10) and Vibrio vulnificus sepsis group (B group, n =50). Sepsis model was made in rats with subcutaneous injection of Vibrio vulnificus with concentration of 6 × 108 cfu/ml in dose of 0. 1 ml/100 g into left lower limb.The rats of group B were sacrificed 1 h, 6 h, 12 h, 24 h and 48 h after infection for taking lung tissues to detect the water content of lung and to observe the histopathological changes in lung under light microscope.The expression of HMGB1 mRNA and the level of HMGB1 protein in the lungs were detected by RT-PCR and Western blot, respectively. Data were analysed with ANOVA and LSD method for comparison between groups, and P ＜0.05 was considered statistically significant. Results Compared with the group A (0.652±0. 177), the expressions of HMGB1 mRNA in lung of rats of group B were significantly higher in 12 hours (1. 161 ±0.358, P=0.013), 24 hours (1.679 ±0.235, P =0.000) and 48 hours (1.258 ±0.274, P=0.004) and reached the peak in 24 h. Compared with group A (0.594 ±0. 190), the level of HMGB1 protein in rats of group B 6 h after infection ( 1. 408 ± 0. 567, P = 0. 026) was significantly increased (P＜0.05), and it reached peak in 24 h (2.415 ± 1.064, P =0.000) after infection. Compared with group A (0.699 ± 0.054), the lung water contents in rats of group B were significantly increased in 6 h (0.759±0.030, P=0.001), in 12 h (0.767 ±0.023, P =0.000), in 24 h (0.771 ±0.043, P=0.000) and in 48 h (0.789 ±0.137, P=0.000) after infection. Compared with group A, the pathological changes in the lung of rats in group B showed clearly marked pulmonary vascular congestion, interstitial edema and inflammatory cell infiltration, and those changes became more and more serious until alveolar sacs entirely collapsed and the boundaries of the alveolar septa could not be clearly identified in 48 h. Conclusions Vibrio vulnificus sepsis leads to the lung injury of infected rats, and the increase in the expression of HMGB1 mRNA in lung might be one of the mechanisms of lung injury in rats with Vibrio vulnificus sepsis.