Objective:To observe the effect of partial liquid ventilation (PLV) on gas exchange,pulmonary compliance and blood dynamics in the pulmonary edema rabbits after seawater drowning(PE-SWD). Methods:Totally 42 healthy male New Zealand rabbits were randomly allocated into 3 groups (n=14);each group was treated with various methods after PE-SWD model was established:PE group with no treatment,CMV group with conventional mechanical ventilation and positive end-expiratory pressure(PEEP),PLV group with CMV,PEEP and FDC.After being treated for 3 h,blood gases,pulmonary mechanics,hemodynamics,pulmonary histopathology and survival time were observed.Results:In PLV group,PaO 2 increased obviously and reached (262.84?64.33) mmHg (1 mmHg=0.133 kPa), airway pressure decreased by 23.4%,pulmonary static compliance increased by 31.3% and TNF-? decreased by 24.9%.HR and MAP showed no significant changes after treatment compared with PE group.Conclusion: Pulmonary gas exchange and compliance are improved significantly during PLV in the PE-SWD rabbits without obvious influence on hemodynamic status, although the survival time of PLV group is less than that of CMV group.

BACKGROUND: Liver inflammation is the main cause of severe liver diseases, including liver fibrosis, steatohepatitis, cirrhosis and hepatocellular carcinoma. Cell therapy topics are receiving increasingly more attention. The therapeutic applications of mesenchymal stem cells (MSC) have become one of the most discussed issues. While other stem cells have therapeutic effects, they have only one or two clinical applications. MSCs are responsible for repairing a variety of tissue injuries. Moreover, MSCs could be derived from several sources, including adipose tissue. MSCs are usually more abundant and easier to obtain compared to other stem cells. METHODS: To prove the concept that MSCs have homing ability to the injured tissue and assist in tissue repair, we examined the effects of intravenous injected adipose-derived mesenchymal stem cells (ADSCs) in a N-nitrosodiethylamine (DEN)-induced liver injury rat model. RESULTS: The significant repairing ability of ADSCs was observed. The levels of fibrosis, apoptosis, and tumorigenesis in the DEN-injured liver tissues all decreased after ADSC treatment. Furthermore, to enhance the therapeutic effects of ADSCs, we pretreated them with L-theanine, which promotes the hepatocyte growth factor secretion of ADSC, and therefore improved the healing effects on injured liver tissue. CONCLUSION ADSCs, especially L-theanine-pretreated ADSCs, have anti-inflammation, anti-apoptosis, and anti-tumorigenesis effects on the N-nitrosodiethylamine-induced liver injury rat model.

Human brain natriuretic peptide (hBNP) was used clinically for the treatment of acute decompensated congestive heart failure. In this paper, hBNP was expressed as a fusion protein with a histidine tag and Ssp dnaB mini-intein which was capable of self-cleavage. After affinity chromatography with Ni-Sepharose and renaturation, the fusion protein was enriched with CM-cellulose. Ssp dnaB mini-intein mediated peptide-bond hydrolysis was triggered by shifting the pH and temperature in the CM-cellulose column, which let to the release of hBNP from the fusion protein and the separation of hBNP from His-DnaB. The hBNP sample was further purified by C4 reverse phase HPLC, and 2.8mg of the peptide with homogeneity of 97% was obtained from one liter of culture medium. The biological activity was assayed in vitro, which indicated that hBNP had a potent vasodilatory effect on rabbit aortic strips with an EC50 of 1.94 x 10(-6) mg/mL.
Animals ; DnaB Helicases ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Inteins ; genetics ; Natriuretic Peptide, Brain ; biosynthesis ; genetics ; Protein Engineering ; methods ; Protein Splicing ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology

The mechanisms leading to the occurrence of seizures in humans are still poorly understood. It is widely accepted, however, that the process of seizure generation is closely associated with an abnormal synchronization of neurons. In order to investigate this process, we have studied synchronization between different regions of the brain from intracranial EEG recordings of Pilocarpine-induced epileptic rat by Synchronization likelihood (SL). It was found that during the state of transition from non-epileptiform discharges to continuous-epileptiform discharges, synchronization between left-ECoG and left-EHG was significantly strengthened, and similar change had taken place between right-ECoG and right-EHGd; left-ECoG; and right-ECoG and left-EHG and right-EHG (P < 0.05). During the state of transition from continuous-epileptiform discharges to period-epileptiform discharges, the synchronization was significantly weakened between left-ECoG and left-EHG, and similar change was noted between left-EHG and right-EHG (P < 0.01). The results showed that SL might be used to assess the dynamics of synchronization and it might be helpful to understanding the mechanisms involving the origin of epileptiform discharge.
Animals ; Cerebral Cortex ; physiopathology ; Electroencephalography ; methods ; Epilepsy ; chemically induced ; physiopathology ; Hippocampus ; physiopathology ; Male ; Pilocarpine ; Rats ; Rats, Sprague-Dawley ; Signal Processing, Computer-Assisted

It was constructed that a genomic DNA library from Lactobacillus sp. MD-1 yielding D, L-lactic acid. The gene encoding L-lactate dehydrogenase (L-LDH) was cloned from the genomic library of strain MD-1 by complementation in E. coli FMJ144 which was lactate dehydrogenase and pyruvate-formate lyase double defective mutant. The nucleotide sequence of the ldhL gene predicted a protein of 316 amino acid starting with ATG. The putative molecular weight of the L-LDH amino acid sequence was 33.84kD. A putative typical promoter (-35 and -10 boxes) had been observed in the 5' noncoding region. An rho-independent transcriptional terminator has been observed in the 3' noncoding region. Three highly conserved regions (Gly13 approximately Asp50, Asp73 approximately Ileul00 and Asn123 approximately Arg154) with several conserved residues had been identified. Gly13 approximately Asp50 was NADH-binding site domain. Asp73 approximately Ileu100 and Asn123 approximately Arg154 were reported to be the active site domains. The ldhL and the L-LDH of Lactobacillus sp. MD-1 showed the low identity and similarity with other Lactobacilli, and the highest percentage were 61.9% and 68.9% respectively. All the above indicated this gene is a novel ldhL.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; L-Lactate Dehydrogenase ; chemistry ; genetics ; physiology ; Lactobacillus ; genetics ; Molecular Sequence Data

In order to study the signal pathway secreting type Ⅰ interferon in porcine alveolar macrophages (PAMs) infected with porcine circovirus type 2 (PCV2), the protein and the mRNA expression levels of cGAS/STING pathways were analyzed by ELISA, Western blotting and quantitative reverse transcriptase PCR in PAMs infected with PCV2. In addition, the roles of cGAS, STING, TBK1 and NF-κB/P65 in the generation of type I interferon (IFN-I) from PAMs were analyzed by using the cGAS and STING specific siRNA, inhibitors BX795 and BAY 11-7082. The results showed that the expression levels of IFN-I increased significantly at 48 h after infection with PCV2 (P<0.05), the mRNA expression levels of cGAS increased significantly at 48 h and 72 h after infection (P<0.01), the mRNA expression levels of STING increased significantly at 72 h after infection (P<0.01), and the mRNA expression levels of TBK1 and IRF3 increased at 48 h after infection (P<0.01). The protein expression levels of STING, TBK1 and IRF3 in PAMs infected with PCV2 were increased, the content of NF-κB/p65 was decreased, and the nuclear entry of NF-κB/p65 and IRF3 was promoted. After knocking down cGAS or STING expression by siRNA, the expression level of IFN-I was significantly decreased after PCV2 infection for 48 h (P<0.01). BX795 and BAY 11-7082 inhibitors were used to inhibit the expression of IRF3 and NF-κB, the concentration of IFN-I in BX795-treated group was significantly reduced than that of the PCV2 group (P<0.01), while no significant difference was observed between the BAY 11-7028 group and the PCV2 group. The results showed that PAMs infected with PCV2 induced IFN-I secretion through the cGAS/STING/TBK1/IRF3 signaling pathway.
Animals ; Cells, Cultured ; Circovirus ; Interferon Type I/genetics* ; Macrophages, Alveolar/virology* ; Membrane Proteins/metabolism* ; Nucleotidyltransferases/metabolism* ; Signal Transduction ; Swine

DraIII is a type IIP restriction endonucleases (REases) that recognizes and creates a double strand break within the gapped palindromic sequence CAC↑NNN↓GTG of double-stranded DNA (↑ indicates nicking on the bottom strand; ↓ indicates nicking on the top strand). However, wild type DraIII shows significant star activity. In this study, it was found that the prominent star site is CAT↑GTT↓GTG, consisting of a star 5' half (CAT) and a canonical 3' half (GTG). DraIII nicks the 3' canonical half site at a faster rate than the 5' star half site, in contrast to the similar rate with the canonical full site. The crystal structure of the DraIII protein was solved. It indicated, as supported by mutagenesis, that DraIII possesses a ββα-metal HNH active site. The structure revealed extensive intra-molecular interactions between the N-terminal domain and the C-terminal domain containing the HNH active site. Disruptions of these interactions through site-directed mutagenesis drastically increased cleavage fidelity. The understanding of fidelity mechanisms will enable generation of high fidelity REases.
Amino Acid Sequence ; Base Sequence ; Calorimetry, Differential Scanning ; Catalytic Domain ; Crystallography, X-Ray ; DNA ; metabolism ; DNA Cleavage ; Deoxyribonucleases, Type II Site-Specific ; chemistry ; genetics ; metabolism ; Escherichia coli ; metabolism ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Substrate Specificity

Arteries in vivo are subjected to lumen pressure, shear flow and axial tension due to surrounding tissue tethering. The axial stress affects arterial function including its response to pressure and flow. While the effects of blood pressure and shear flow are well documented, the effects of axial tension on vascular remodeling have just gradually gained attention recently. This review summarizes the results on the observation of the axial tension in arteries and responses of arteries to elevation and reduction of the axial stress. It is concluded that the axial tension in arteries plays an important role in regulating normal arterial function and tissue remodeling and adaptation and disease development. Research on vascular remodeling under axial tension shall strengthen the understanding of normal physiological functions and pathological changes of the arteries.

OBJECTIVETo establish the induction method and culture system of hairy roots of Panax japonicus, and determine ginsenoside Re contents.

METHODHairy roots of P. japonicus was obtained through infecting pre-incubated terrestrial stem with the Agrobacterium tumefaciens strain C58C1; its enlarging culture was carried out on the 1/2MS medium, and growing characteristics were measured. The transformation of T-DNA was examined by PCR and ginsenoside Re content was determined by HPLC.

RESULTA. tumefaciens strain C58C1 could make terrestrial stem of P. japonicus bring about hairy roots, the max inductivity was 90% when infecting for 25 min. The PCR examination result showed that rolB genes could be inserted into the hair roots of P. japonicus. All hairy roots dould synthesize ginsenoside Re, among them, the max content was PJ8 with 60. 26 mg x g(-1).

CONCLUSIONIt was reported for the first time that the induction method and culture system of hairy roots of P. japonicus were established successfully, which provided a foundation for producing high content ginsenoside Re through culturing the hairy roots of P. japonicus.

Agrobacterium tumefaciens ; genetics ; physiology ; Cells, Cultured ; Culture Techniques ; methods ; Ginsenosides ; biosynthesis ; Panax ; genetics ; growth & development ; metabolism ; microbiology ; Plant Roots ; genetics ; growth & development ; metabolism ; microbiology

Both phytase and endoglucanase are additives in feed for mono-gastric animal known for their effects. Recombinant vector pPICZalpha-EG was constructed and transformed to GS115-phyA, a Pichia pastoris strain that had integrated with phytase gene, generating GS115-phyA-EG. Both phytase and endoglucanase activities in the supernatant were determined after methanol induction of GS115-phyA-EG. Phytase and endoglucanase activity reached 39.4% and 56.2% activity compared to GS115-phyA and GS115-EG, respectively. Properties of the mixed enzyme suggest that the optimal temperature and pH value be 55 degrees C and 5.5 respectively. Both phytase and endoglucanase showed greater than 80% activity across temperature ranges 45 degrees C to 55 degrees C and pH ranges 4.5 to 5.5. Expressing more than one enzyme in one system could save time and money during induced expression, and the mixed enzyme might apply for treating forge before feeding with poultry.
6-Phytase ; biosynthesis ; genetics ; Cellulase ; biosynthesis ; genetics ; Genetic Vectors ; Pichia ; enzymology ; genetics ; Recombinant Proteins ; biosynthesis ; genetics