The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Tripterygium wilfordii has exihibited multiple pharmacological activities, such as anti-inflammatory, immune modulation, anti-tumor and anti-fertility. T. wilfordii have been used for the therapy of inflammation and autoimmune diseases including rheumatoid arthritis, immune complex nephritis and systemic lupus erythematosus clinically. However, it is well known that T. wilfordii has small margin between the therapeutic and toxic doses and could cause serious injury on digestive, reproductive and urogenital systems. Among all the organs, liver is one of the most remarkable targets of T. wilfordii-induced toxicities, and the damage is more serious than others. It is generally accepted that T. wilfordii-induced liver injury is a result of the combined effects of toxic elements of T. wilfordii. It is reported in several studies that the mechanism of T. wilfordii-induced liver injury may be related to lipid peroxidation, cell apoptosis and immune damage, and so on. Licorice is one of the most commonly used Chinese herbal medicine, with effects of heat- clearing and detoxicating, anti-inflammatory and hepatoprotective, reconciling various drugs, and so on. Licorice often accompany T. wilfordii in clinical application which can significantly reduce the liver injury induced by T. wilfordii. The attenuated effect is exact, but the mechanism is still a lack of in-depth study. This paper reviews the studies on T. wilfordii-induced liver injury and the related mechanism as well as licorice and other traditional Chinese medicine accompany T. wilfordii to reduce the injury in recent years, so as to provide reference for related research in the future.
Animals ; Chemical and Drug Induced Liver Injury ; etiology ; prevention & control ; Glycyrrhiza ; Humans ; Inactivation, Metabolic ; Medicine, Chinese Traditional ; Tripterygium

OBJECTIVETo compare the event-related potentials (ERPs) waves of verbs and prepositions in the brain.

METHODSWe recorded ERPs in the brain while participants judged the legality of the collocation for verbs and prepositions.

RESULTSBoth verbs and prepositions elicited a negativity at the frontal site in 230-330 ms and 350-500 ms window. No difference was seen in 230-330 ms and 350-500 ms window; In difference waves, a negativity was elicited in the left and right hemisphere at about 270-400 ms and 470-600 ms window for both open and closed-class words.

CONCLUSIONThese may demonstrate that prepositions in modern Chinese are probably not a separate class from verbs and that N280 may be not a specific component for only prepositions (or closed-class words).

Adult ; Asian Continental Ancestry Group ; Brain ; physiology ; Electroencephalography ; Evoked Potentials ; physiology ; Female ; Humans ; Language ; Male ; Photic Stimulation ; Reading

Objective To investigate the distribution and antimicrobial susceptibility of pathogens causing maxillofacial infection in patients,and provide evidence for rational use of antimicrobial agents in clinical practice.Methods Specimens of patients infected with oral and maxillofacial infection in a hospital between January 2012 and December 2016 were performed microbial culture,pathogens were identified and performed antimicrobial susceptibility testing,distribution of pathogens and antimicrobial resistance were analyzed.Results Of 882 patients with maxillofacial infection,male and female accounted for 32.20％ and 67.80％ respectively;35.38％ and 32.65％ of patients aged ～40 years and ～60 years respectively;a total of 145 strains were isolated,88(60.69 ％) of which were gram-negative bacteria,mainly Klebsiella pneumoniae and Pseudomonas aeruginosa;56 (38.62 ％) of which were gram-positive bacteria,mainly Staphylococcus aureus.Resistance rates of Klebsiella pneumoniae to 16 kinds of an timicrobial agents were a11＜50％,resistance rates to imipenem and meropenem were the lowest,both were 3.45％,1 1 strains were extended-spectrum β-lactamases-producing strains;resistance rates of Pseudomonas aeruginosa to 10 kinds of antimicrobial agents were all＜40％.Staphylococcus aureus was susceptible to tigecycline,linezolid,and vancomycin,resistance rate to penicillin was the highest (66.67％),resistance rate to oxacillin was 20.83％.Conclusion The main pathogens causing oral and maxillofacial infection are gram negative bacteria,different pathogens have different antimicrobial resistance,antimicrobial agents should be used rationally during clinical treatment according to antimicrobial susceptibility testing result.

OBJECTIVETo investigate the relationship between the SNP rs11614913 on miR196a-2 gene and the treatment effects of Peg-IFN-a plus Ribavirin on chronic hepatitis C patients.

METHODSThe total 139 patients of chronic hepatitis C infection who received the treatment of Peg-IFN-alpha-2a or Peg-IFN-alpha-2b plus Ribavirin were enrolled in this study. The patients were divided into two groups: sustained virological response (SVR) (n = 82) group and non virological response (NVR) or recurrence (n = 57) group. Blood samples were collected and chromosomal DNA was extracted. The miR-196a-2 polymorphism was determined with the method of polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP).

RESULTSIn our study, there was statistically association between miR-196a-2 polymorphism and the antiviral therapy efficacy of hepatitis C patients. There was statistically significance in the CT genotype and the TT genotype of miR-196a-2 between the two groups [P = 0.009, A = 2.924 (1.285 -6.652)]. There was statistically significance in the CC genotype and the TT genotype between the two groups [P = 0.036, A = 3.091(1.052 -9.078)]. There was statistically significance in the C allele and the T allele between the two groups [P = 0.036, A = 3.091 (1.052 - 9.078)].

CONCLUSIONThese findings suggested that the rs11614913 SNP in miR - 196a-2 be associated with the antiviral therapy efficacy of hepatitis C patients, and the TT genotype or T alleles be associated with the SVR while the CC genotype or C allele could be related to the NVR or recurrence.

Adolescent ; Adult ; Aged ; Alleles ; Antiviral Agents ; therapeutic use ; Hepatitis C, Chronic ; drug therapy ; genetics ; Humans ; Interferon-alpha ; therapeutic use ; MicroRNAs ; genetics ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Polymorphism, Genetic ; Recombinant Proteins ; Ribavirin ; therapeutic use ; Young Adult

OBJECTIVETo explore the relationship between the expression of Survivin and Ki67 with prognosis of pancreatic endocrine tumors (PETs).

METHODSImmunohistochemistry for Survivin and Ki67 was performed in 25 cases of normal pancreatic tissues and 81 cases of PETs by tissue microarrays and to observe the expression and evaluate the relationship with prognosis.

RESULTS(1)The expression of Survivin and Ki67 in PETs was significantly higher than that in normal pancreatic tissues (P <0.01); (2)The expression of Survivin and Ki67 in PETs was correlated with tissue grading and the TNM-staging (P < 0.05), but not related with tumor size, location and functional status. In addition, the expression of nuclear Survivin was association with lymph node metastasis (P < 0.05). (3)The high expression of Ki67 was related with the expression of nuclear Survivin, but not related with the expression of cytoplasmic Survivin.

CONCLUSIONSurvivin and Ki67 were both expressed in PETs, which were closely related to the clinical pathological characteristics. They could be used as new indicators in the evaluation of prognosis of PETs. The expression of Survivin in nucleus had more diagnostic significance than that in cytoplasm, and that could be highly correlated with lymph node metastasis, which would be used as a new marker of poor prognosis.

Biomarkers, Tumor ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; metabolism ; Ki-67 Antigen ; metabolism ; Lymphatic Metastasis ; Neoplasm Staging ; Pancreatic Neoplasms ; diagnosis ; Prognosis

Spontaneous transient outward currents (STOCs) play an important role in the myogenic regulation of small artery tone, such as coronary artery. In the present study, we investigated the electrophysiological properties and the regulation of STOCs in vascular smooth muscle cells (VSMCs) of porcine coronary artery by perforated patch-clamp technique. Our data showed that STOCs were dependent on voltage and extracellular calcium and they were highly variable in amplitudes and frequencies. STOCs superimposed stochastically onto whole-cell K(+) currents induced by step and ramp protocols. STOCs were completely abolished by ChTX [inhibitor of large-conductance Ca(2+)-activated potassium (BK(Ca)) channels], removal of extracellular Ca(2+), or addition of ryanodine (50 mumol/L) respectively. In contrast, CdCl2 and verapamil, inhibitors of voltage-dependent L-type Ca(2+) channels, had little effect on STOCs. Caffeine (5 mmol/L) transiently increased STOCs (hump), followed by a temporary inhibition. Ca(2+) ionophore A23187 increased both amplitude and frequency of STOCs. Na(+) ionophore monensin increased the frequency of STOCs. STOCs were strongly inhibited by KB-R7943, a selective inhibitor of the reverse mode of the Na(+)/Ca(2+) exchanger. Based on these observations, we conclude that STOCs are mediated by BK(Ca) channels. The generation and activation of STOCs depend upon Ca(2+) influx through Na(+)/Ca(2+) exchange and release of Ca(2+) from sarcoplasmic reticulum (SR) via ryanodine receptors. This suggests that Na(+)/Ca(2+) exchange determines calcium store refilling. Recycling of entering Ca(2+) from superficial SR may locally elevate Ca(2+) concentration at the plasma membrane, thereby activating BK(Ca) channels and then initiating STOCs.
Animals ; Coronary Vessels ; cytology ; physiology ; Electrophysiological Phenomena ; physiology ; Muscle, Smooth, Vascular ; cytology ; physiology ; Myocytes, Smooth Muscle ; cytology ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; physiology ; Sodium-Calcium Exchanger ; physiology ; Swine

Actinobacillus pleuropneumoniae ; drug effects ; genetics ; Animals ; Bacterial Proteins ; genetics ; Blotting, Western ; Drug Resistance, Bacterial ; genetics ; Green Fluorescent Proteins ; genetics ; Hemolysin Proteins ; genetics ; Hemolysis ; Mice ; Mutation ; Polymerase Chain Reaction

BACKGROUNDThe etiologic agent of severe acute respiratory syndrome (SARS) has been confirmed to be a novel coronavirus (CoV), namely SARS-CoV. Developing safe and effective SARS-CoV vaccines is essential for us to prevent the possible reemergence of its epidemic. Previous experiences indicate that inactivated vaccine is conventional and more hopeful to be successfully developed. Immunogenicity evaluation of an experimental inactivated SARS-CoV vaccine in rabbits was conducted and reported in this paper.

METHODSThe large-scale cultured SARS-CoV F69 strain was inactivated with 0.4% formaldehyde and purified, then used as the immunogen combined with Freund's adjuvant. Eight adult New Zealand rabbits were immunized four times with this experimental inactivated vaccine. Twelve sets of rabbit serum were sampled from the third day to the seventy-fourth day after the first vaccination. The titers of specific anti-SARS-CoV IgG antibody were determined by indirect enzyme-linked immunosorbent assay, and the neutralizing antibody titers were detected with micro-cytopathic effect neutralization test.

RESULTSRapid and potent humoral immune responses were induced by the inactivated SARS-CoV vaccine in all the eight test rabbits. Titers of both specific IgG antibody and neutralizing antibody peaked at about six weeks after first vaccination, with the maximum value of 1:81 920 and 1:20 480, respectively. After that, serum antibody levels remained at a plateau or had a slight decrease, though two boosters were given in the succedent 4 to 5 weeks. Cross neutralization response existed between SARS-CoV F69 strain and Z2-Y3 strain.

CONCLUSIONSThe inactivated SARS-CoV vaccine made from F69 strain owns strong immunogenicity, and the cross neutralization response between the two different SARS-CoV strains gives a hint of the similar neutralizing epitopes, which provide stable bases for the development of inactivated SARS-CoV vaccines.

Animals ; Antibodies, Viral ; blood ; Immunoglobulin G ; blood ; Neutralization Tests ; Rabbits ; SARS Virus ; immunology ; Vaccines, Inactivated ; immunology ; Viral Vaccines ; immunology

Objectives: This study aimed to observe the change of arachidonic acid-induced platelet aggregation rate (AA-Ag) and short-term adverse reactions after taking 50 or 100 mg/d aspirin(enteric-coated sustained-release formulation) or 100 mg/d aspirin (enteric-coated aspirin tablet)in the elderly Chinese population (aged 60 years or older). Methods: A total of 1 194 participants aged 60 or older, who should be recommended to take aspirin therapy due to medical reasons, were recruited and randomly assigned into three groups to receive enteric-coated sustained-release aspirin tablet (50 mg, once daily, group A), or 100 mg, once daily (group B) or enteric-coated aspirin tablet 100 mg once daily (group C), respectively. AA-Ag was measured after (14±3)days of aspirin treatment. Adverse events and bleeding events were recorded during the (28±3)days of follow-up. Results: The AA-Ag in group A (n=347), B (n=338) and C (n=332) post 14-day aspirin therapy were 6.65 (4.03,10.84)%, 5.89(3.22,10.03) % and 6.00(3.68,10.09) %, respectively (P>0.05). During the 28 days follow-up, the adverse events rate of group A (n=388), B (n=387) and C (n=385) was 3.87%,3.36%, and 7.95%, and the mild bleeding events rate was 3.09%, 2.33%, and 6.23%, respectively. Adverse events rate and mild bleeding events rate were significantly higher in group C than in group A and B (P<0.05). Conclusions: Compared with 100 mg-dose aspirin, 50 mg-dose aspirin achieves similar anti-platelet aggregation effect in this elderly Chinese population. The short-term adverse events and mild bleeding risk of aspirin with enteric-coated sustained-release formulation were fewer than that of enteric-coated formulation.