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.

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.

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 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 potential role and changes of CD14, TNF-ct and IL-10 in liver in Vibrio vulnificus septic rats, and detect the intervention effects of eefoperazone sodium combined with levofloxacin. Methods To make Vibrio vulnificus sepsis model (VV group) and drug intervention model (AA group) in rats, the expression of CD14, TNF-α, IL-10 in liver were detected by RT-PCR. Results Compared with normal control (NC) group,CD14 mRNA and TNF-α mRNA expression increased markedly at 2, 6, 9, 12, 16 h in VV group (P<0.05), IL-10 mRNA raised greatly at 9, 12, 16 in VV group (P<0.05). CD14 mRNA expression was also rised in AA group at 9 h(P<0.05). TNF-α mRNA at 9, 12 h and IL-10 mRNA at 9, 12, 16 h in the AA group increased (P<0.05). Compared with VV groups, CD14 mRNA expression diminished greatly at 9, 12, 16 h in AA group (P <0.05), TNF-α mRNA and IL-10 mRNA diminished in the AA group at 16 h(P<0.05). Conclusion The treatment with cefoperazone sodium and levofloxacin may reduce expression of CD14, TNF-α and IL-10 in liver of rats with VV sepsis, it may inhibit the level of pro-inflammatory/anti-inflammatory cytokines, thereby regulating the balance of the inflammatory response in VV sepsis.

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.

Objective To investigate the expressions of TF mRNA and TFPI mRNA of liver in rats with Vibrio vulnificus sepsis and to assess the interventional effects of cefoperazone sodium along with levofloxacin lac-tate. Method One hundred and ten male SD rats were divided (random number) into normal control group (NC group, n = 10), Vibrio vulnificus sepsis group (VV group, five subgroups n = 10 in each), drug intervention model (AA group, five subgroups n = 10 in each). The Vibrio vulnificus sepsis models and drug intervention models of rat were made. The reverse transcription polymerase chain reaction (RT-PCR) assay was employed for the measurement of TF mRNA and TFPI mRNA. ANOVA and t-test performed with SPSS version 12.0 software. Results Compared with NC, the expressions of TF mRNA in liver increased markedly 2 h,6 h, 12 h and 16 h af-termodeling in VV groups (P<0.05), and reached peak 6 hours after modeling. The expressions of TF mRNA in liver of rats in AA groups were much higher than those in NC group 9 h and 12 h after modeling (P<0.05). The expressions of TFPI mRNA in liver of rats in VV groups and AA groups were not significantly different to those in NC group (P>0.05). Compared with VV groups, the expressions of TF mRNA in liver of rats in AA groups were greatly lowered 9 hours after administration of bactericide (P<0.05), and the expressions of TFPI mRNA in liver of rats in AA groups were significantly higher 12h and 16 h after intervention (P<0.05). Conclusions There is a obvious imbalance between coagulation and anticoagulation functions of circulation system during Vibrio vulnificus sepsis, and the imbalance can be corrected gradually after treatment with antibacterial agents.

Objective To study epidemiology, clinical findings, diagnosis and treatment of sepsis caused by Vibrio vulnificus. Method Patientss with Vibrio vulnificus sepsis were collected from 1995 to 2008. The medical records including epidemiological and clinical data were analyzed. Results The male-to-female ratio of 34cases was 4.7:1 and 76. 5% of these patients suffered from chronic liver disease. Most patients occurred from April to October with signs of abrupt fever, characteristic cutaneous lesions, hypotension and progressive multiple organ disfunction syndrome (MODS). The mortality was over 47.1% . The criteria proposed for early diagnosis of Vibrio vulnificus sepsis were abrupt onset with fever during the period from April to November, characteristic cutaneous lesions, such as the most commonly occurred haemorrhagic bullae on the extremities or even extensive necrosis of skin and muscular tissue, progressive hypotension or shock accompanied by MODS, pre-existing liver disease or chronic abuse of alcohol, and consumption of raw seafood or exposure to seawater within 12 week. Early administration of the third-generation cephalosporins with the quinolones in full dosage, aggressive wound debridement,appropriate dermoplasty and supportive care contribute to a better outcome. Conclusions Vibrio vulnificus sepsis progresses rapidly with high mortality. Early diagnosis, rapid treatment with prompt antibiotics and aggressive surgery treatment are very important to improve the outcome.