DC-12 is a Bacillus strain with strong fibrinolytic activity was screened from Douchi.A novel breeding technology——whole genome shuffling was applied to enhance the production of fibrinolytic enzyme.First of all,a candidate mutant library was constructed by treating DC-12 with ultraviolet rays andhNO2,respectively.Based on studying the optimum conditions for protoplast preparation and regeneration of DC-12,multi-parental whole genome shuffling with 4 strains was conceded by protoplasts electrical fusion,then combines with the screening method of double-inactive protoplast,two strains withhigher yield of fibrinolytic enzyme were obtained effectively,which can descend stably after five generations,and their enzyme production reached 2710 IU/ml,which was increased by 4~5 fold compared with the mutants,respectively.

Objective To investigate the effect of 20‐HETE on the isolated myocardial ischemia reperfusion injury and to ex‐plore its underlying mechanisms .Methods Experiments were performed in isolated rat hearts subjected to 35 min of ischemia fol‐lowed by 40 min of reperfusion in Langendorff preparations .HET0016 (1 μmol/L) and various concentrations (10 ,30 or 50 nmol/L) of 20‐HETE were infused 10 min before the onset of ischemia and throughout the reperfusion period .Cardiac hemodynamic changes and myocardial contractility were continuously recorded with the Powerlab /8P system .Myocardial infarct size was meas‐ured by TTC staining .The level of ROS and the protein carbonyl content were determined by DHE fuorescence and DNPH method , respectively .Results Perfusion with HET0016 significantly improved myocardial ischemia reperfusion injury reduction in cardiac contractility ,after inhibited the production of 20‐HETE significantly reduced the occurrence of myocardial infarction area (P<0 .05) ,but exogenous join 20‐HETE aggravated I/R‐induced myocardial injury (P<0 .05) .Myocardial ischemia reperfusion injury significantly increased production of ROS and oxidative stress ,both of which were significantly inhibited by HET 0016 and enhanced by 20‐HETE administration(P< 0 .05) .Conclusion 20‐HETE stimulates ROS production and enhance protein carbonylation , which aggravates myocardial ischemia reperfusion injury .

To study the influence of foreign plasmid DNA on spleen metabolism in immune-stimulated mice via the gastrointestinal tract. Mice were oral administered by pipette with 200?g of plasmid pcDNA3 and spleen were isolated at 4h and 18h after oral administration. Total RNA were extracted from spleen and spleen gene expression profile of Balb/c mice was analyzed by using Affymetrix oligonucleotide genechip after oral plasmid pcDNA3 administration. Functional cluster analysis was conducted by Genmapp and MAPPFinder software. By functional cluster analyzing the genes which were up-regulated more than twofolds, Genmapp results showed that plenty of metabolic pathway were induced in spleen after oral administration of plasmid pcDNA3. These metabolic process included purine metabolism, pyrimidine metabolism, protein synthesis, cholesterol synthesis, fatty acid synthesis, Glycolysis, TCA cycle and mitochondria oxidative phosphorylation pathway. The similar results also took place at 18h after oral administration. The result indicated that foreign plasmid DNA can modulate metabolism process in spleen of mice via the gastrointestinal tract, and may help understand the mechanism of action of foreign plasmid DNA uptaked via the gastrointestinal tract.

Aim To investigate the effects of U50488H,a selective ?-opioid receptor agonist,on the blood pressure in the rats and explore their mechanisms.Methods Heart rate(HR),arterial blood pressure(ABP),left ventricular pressure(LVP),contractive function(+ dp/dt_(max)) and diastolic function(-dp/dt_(max)) were examined in rats.Physiological experimental technique was used to collect urine and to determine the volume of urine output;isolated artery perfusion technique was used to investigate the direct action of U50488H on abdominal aorta of rats.Results HR,ABP,LVP and ?dp/dt_(max) in rats were decreased with the administration of U50488H;the urine volume increased significantly with the administration of U50488H.U50488H induced a dose-dependent vasodilation in the aortic artery.These effects of U50488H were totally abolished by nor-BNI,a selective ?-opioid receptor antagonist.Conclusion Stimulation of ?-opioid receptor with U50488H depresses the blood pressure mainly by reducing the strength of cardiac muscle,enhancing the urine volume and relaxing the vessel.

AIM: To activate hepatic stellate cells (HSC) in vitro in precision-cut liver slices (PCLS) stimulated by acetaldehyde for studying and screening anti-fibrotic drugs. METHODS: PCLS were prepared by the vibratome, and incubated with 700 ?m?L~-1 acetaldehyde for 0, 2, 4 and 6 h. The medium and homogenate were retained to determine glutathione S-transferase (GST) activity, lactate dehydrogenase (LDH) leakage and hydroxyproline (Hyp) content. PCLS were prepared for paraffin sections. The expression of ?-smooth muscle actin (?-SMA) was evaluated by immunohistochemistry, and the result was analyzed with image analysis software. RESULTS: The leakages of GST and LDH were increased significantly compared with those in 0 h group (P

Adult ; Anesthetics, Inhalation ; administration & dosage ; Desflurane ; Electroencephalography ; methods ; Female ; Humans ; Isoflurane ; administration & dosage ; analogs & derivatives ; Male ; Middle Aged ; Monitoring, Intraoperative ; Nitrous Oxide ; administration & dosage

To study the identification of Gentianaceae Mongolian medicine Digeda with spectroscopy techniques, near infrared spectroscopy and differential scanning calorimetry techniques were applied to study on the identification of 4 kinds of Gentianaceae Mongolian medicine Digeda, and characteristic spectrums obtained were systematically analyzed. In NIR study, the four species of Digeda exist some differences in 4 250-4 400 cm(-1) and 5 650-5 800 cm(-1) of one-dimensional spectra, and show significant differences in 4 100- 4 400 cm(-1), 4 401-4 900 cm(-1) and 5 400-5 800 cm(-1) of the second derivative spectra. DSC curves of them present distinct topological pattern, characteristic peak and peak temperature. Using near infrared spectroscopy and differential scanning calorimetry analysis can realize efficient and accurate identification of four kinds of Mongolian medicine Digeda, and provide scientific basis for the efficient and accurate identification of other Gentianaceae Mongolian medicine Digeda.
Calorimetry, Differential Scanning ; methods ; China ; Gentianaceae ; chemistry ; classification ; Medicine, Mongolian Traditional ; Spectroscopy, Near-Infrared ; methods

Objective To develop the hypothesis ‘saturated or non-saturated cytotoxicity model' and explain the various phenomena of antibody mediated immunoresponses in recipients,including rejection and accommodation.Methods The imitating complement dependent cytotoxicity.The threshold set to identify as saturated or non-saturated cytotoxicity depends on antigen-antibody complex(R)whether or not above lethal number(D)in effective time.Feasibility of the hypothesis was examined through explaining various phenomena mediated by anti-donor antibodies,especially some contradictory phenomena.Results Hyperacute rejection,accelerated rejection and acute rejection could be well explained by saturated cytotoxicity.Accommodation of ABO imcompatible transplantion,de novo antibody induced injury,change of protein profile,and C4d deposition in graft could be well elucidated by the hypothesis.Conclusion The hypothesis saturated or nonsaturated cytotoxicity model' help to interpret and interconnect various phenomena of antibodies mediated immune response,such as rejection and accommodation.

Objective To investigate the optimal concentration and site for ozoneadministration to relieve neuropathic pain in rats. Methods Twenty-four adult male rats with sciaticnerve injury (SNI) were randomized equally into 4 groups, and received ozone injections for analgesia atdifferent sites, namely the operative site, the sciatic nerve, the L5 spinal nerve or the operative site inaddition to the L5 spinal nerve. Another 4 groups of rats with SNI (n=6) had ozone injections at theconcentrations of 40, 50, 60, or 70 μg/mL, all administered at both the operative site and the L5 spinalnerve. The analgesic effects of the ozone injections were evaluated using paw withdrawal threshold test inall the rats. Results In rats with ozone injection at both the operative field and the L5 spinal nerve, themechanical pain threshold was significantly increased in comparison with that in rats with injections atother sites (P<0.05), but different doses of ozone simultaneously injected at the two sites did not producesignificant differences in the pain threshold (P>0.05). Conclusion Ozone produces obvious analgesiceffects in rats with SNI-induced neuropathic pain when injected at both the operative site and thespinal nerve, and ozone within the concentration range from 40 to 70 μg/mL has similar analgesic effect.

OBJECTIVETo compare the clinical curative effect of thoracolumbar burst fracture treated by the posterior unilateral approach corpectomy fusion screw-rod fixation and anterior corpectomy bone fusion screw plate fixation.

METHODSFrom January 2008 to May 2014,36 cases of thoracolumbar burst fracture underwent operation of decompression, fusion, and internal fixation was retrospective analyzed. Among them, 16 patients were treated through posterior approach as posterior group, including 13 males and 3 females aged from 37 to 62 years old; 9 cases caused by falling injury, 3 cases by traffic accident injury,4 cases by heavy aboved;the injury segment was on T₁₂ in 2 cases, L₁ in 5 cases, L₂ in 7 cases, L₃ in 2 cases; according ASIA grade, 3 cases were grade A, 2 cases were grade B, 2 cases were grade C, 5 cases were grade D, 4 cases were grade E; the time between injury and operation ranged from 5 to 15 days. Other 20 patients were treated through anterior-lateral approach as anterior-lateral group, including 15 males and 5 females with age from 27 to 62 years old; 12 cases caused by falling injury, 4 cases by traffic accident injury, 4 cases by heavy aboved; the injury segment was on T₁₂ in 2 cases, L₁, in 7 cases, L₂ in 9 cases, L₃ in 2 cases; for ASIA grade: 4 cases were grade A, 2 cases were grade B, 4 cases were grade C, 6 cases were grade D, 4 cases were grade E; the time between injury and operation ranged from 4 to 12 days. The operation time, bleeding during operation and postoperative drainage volume were observed in two groups,and the changes of nerve function of ASIA grade, clinical efficacy,improved degree of thoracic and lumbar lordosis,and bony fusion were compared between two groups.

RESULTSAll patients were followed up from 12 to 24 months with an average of (15.8 ± 3.3) months. The operation time, bleeding during operation, and postoperative drainage volume had no significant different between two groups (P > 0.05). As compared with preoperative, ASIA grade of two groups at last follow-up had statistically significantly different (P < 0.01), the neural function of two groups after operation was recovered for different extent. The JOA score of two groups was compared between last follow-up and preoperative, the difference had statistically significant (P < 0.01), the two groups showed good clinical effect. The clinical results of ASIA grade, JOA score and RIS had no significant differences between two groups. All patients of two groups were obtained fusion. Thoracic and lumbar lordosis angle improvement degree had no significant difference between two groups ,it bad significant difference had statistical significance compared with preoperative, the two approaches could effectively restore the spinal sequence.

CONCLUSIONFor patients with thoracolumbar burst fracture just treated by anterior decompression and reconstruction of anterior column, according to the degree of operation performer' skill proficiency and the patient' condition to choose, but for patients must performed the spinal canal decompression anterior and posterior, the three column-reconstruction to required anterior-posterior approach, the posterior unilateral approach corpectomy fusion screw-rod fixation obviously shorten operation time, reduce the operation wound, it is worth the clinical promotion.

Adult ; Decompression, Surgical ; methods ; Female ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Reconstructive Surgical Procedures ; Retrospective Studies ; Spinal Fractures ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; injuries ; surgery