Objective:To enhance the immune efficiency of porcine reproductive and respiratory syndrome virus(PRRSV)DNA vaccine,the ORF5 genes were used as candidate genes to construct the recombinant plasmids CpG-pVAX1-ORF5 by pVAX1 as eukaryotic expression vector.The aim is to analyze the immune responses and the effects of CpG-ODN induced by GP5 recombinant plasmids of PRRSV.Methods:Piglets were immunized with recombinant DNA plasmids which expressed PRRSV GP5 for three inoculations.The PRRSV antibody in serum,the concentration of IL-2 and the lymphocyte proliferation test(MTT) in peripheral blood of vaccinated piglets were detected.The vaccinated pigs were challenged intranasally with PRRSV SD2.Results:GP5 DNA immunization with CpG resulted in the production of both PRRSV antibodies and cellular immune(a significant enhancement of a lymphoproliferative response ).The CpG-pVAX1-ORF5 was showed significantly better protection from the PRRSV challenge compared with the control plasmid.This immune response was characterized by a significantly decreased frequency of viraemia with decrease of 80% compared with the control group.There were little clinical symptoms and protection in some extent from lung damage.Conclusion:The results indicate that CpG-ODN could be used as immune adjuvant of PRRSV DNA vaccine.

Objective To evaluate the significance of serum prealbumin, bilirubin/albumin( B/A )ratio and brainstem auditory evoked potential (BAEP)for early predicting the bilirubinic neurotoxicity caused by early neonatal hyperbilirubinemia. Methods According to the gestational age, birth weight and the intervention criterion of neonatal jaundice, 94 neonates (newborn infants with a age of 1-7days) with hyperbilirubinemia were divided into two groups, one was term neonates with normal BAEP, the other was term neonates with abnormal BAEP. We detected their serum levels of total bilirubine, prealbumin, BAEP and calculated B/A ratio. Result The prealbumin levels was significantly lower and B/A ratio was higher in group of abnormal BAEP than those of control group ( P < 0. 01 ), while the levels of total bilirubin were not significantly different between the two groups ( P > 0. 05 ). In group of abnormal BAEP , there was a negative correlation between the level of serum prealbumin and the B/A ratio (r = -0. 637, P <0. 01 ). Conclution Prealbumin may be a sensitive indicator for early predicting bilirubinic toxicity like B/A ratio and BAEP.

Objective To investigate the characteristic changes in the mRNA and protein of complement 3 in spinal dorsal horn of rats with neuropathic pain,and the role of abnormal activation of complement protein in the mechanism of pain production.Methods Eighty-four healthy male SD rats were divided randomly into seven groups(n=12 for each group):normal control group,sham-operation 1d,3d and 7d groups,and chronic constriction injury of the sciatic nerve(CCI)1d,3d and 7d groups.The left sciatic nerve was ligated loosely in CCI groups,while it was only exposed but not ligated in rats in sham-operation groups.The mechanical and thermal pain thresholds were measured on 1,3 and 7 days after operation,and the mRNA and protein of complement 3 in the dosal horn of the spinal cord were determined respectively by RT-PCR,immunoturbidimetry and immunohistochemistry.Results The mechanical and thermal pain thresholds were observed to be lowered in rats one day after the sciatic nerve ligation,and a state of hyperalgesia was found to be persistent up to 7 days after CCI.This symptom was not observed in sham operation group.The expression of mRNA and protein of complement 3 in spinal dorsal horn were increased on 1,3 and 7 days after CCI.Interestingly,a high expression of mRNA of complement 3 was also observed in rats one day after sham-operation.Both mRNA and protein of complement 3 were not obviously elevated in rats of sham operation 3d,7d groups and normal control group.Conclusion The mRNA and protein of complement 3 in spinal dorsal horn are highly up-regulated in rats with neuropathic pain,suggesting that the characteristic dynamic changes in complement may contribute to the establishment and maintenance of hyperalgesia.

From the prospective of metaethics and normative ethics, we analyze the ethical character and function of medical professional language, and emphasize the influence of respecting the patients'right as well as the role of medical professional language in dealing with doctor - patient relationship.

Objective To study inhibitory activity of anthraquinone in Polygonum Cillinerve(Nakai) Ohwi (anthraquinone in PCO) to nitrosation. Methods Anthraquinone was extracted from Polygonum Cillinerve(Nakai) Ohwi with sulfuric acid and chloroform as solvents. The capability of scavenging sodium nitrite and disconnecting nitrosamine synthesis with anthraquinone in PCO were determined by spectrophotometry under simulated human gastric juice. Results Both the sodium nitrite scavenging rate and the nitrosamine synthesis disconnecting rate showed the positive correlation with the concentration of anthraquinone in PCO. The strongest capability of scavenging sodium nitrite was 53.5%, and the strongest capability of disconnecting nitrosamine synthesis was 71.3%. Conclusion Anthraquinone in PCO had strong capability of scavenging sodium nitrite and disconnecting nitrosamine synthesis.

To prepare a large amount of pure alive Schistosoma japonicum eggs, rabbit was infected with 2000 cercariae and its liver was taken aseptically 38-45 days after infection and homogenized. The homogenate was screened through different sieves(60, 120, 200, 300, 360 meshes per inch respectively), and washed with 1.2% NaCl. The eggs and leftover were then digested with 0.25% trypsin for 2 hours, sieved over 360 meshes per inch and washed with RPMI 1640 medium. The collected eggs reached to (95.1?6.4)% of live eggs, with a high efficiency.

Several virus families have been shown to encode microRNAs (miRNAs), which have roles in the infection and replication of viruses in host cells. These virus-encoded miRNAs are identified in double-stranded DNA virus (dsDNA virus) and in several RNA virus families, but not in single-stranded DNA virus (ssDNA virus). We used a bioinformatics approach based on VMir, miRNAFold and MaturePred software to predict virus-encoded miRNA-like small RNAs from the genome of a ssDNA virus: Aedes aegypti densovirus (AaeDV). Northern blotting and stem-loop reverse transcription-polymerase chain reaction (RT-PCR) were used to detect predicted small RNAs. A miRNA-like small RNA termed "AaeDVMD" was identified by stem-loop RT-PCR from predicted candidates. This is the first report demonstrating that a ssDNA virus can encode miRNA-like small RNAs. These data will aid further exploration of the interaction between the AaeDV and its mosquito host.
Aedes ; virology ; Animals ; Base Sequence ; Computational Biology ; Densovirinae ; chemistry ; genetics ; metabolism ; MicroRNAs ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; RNA, Viral ; chemistry ; genetics ; metabolism

Objective To evaluate the role of membrane-bound complement regulatory protein expression in spinal cord dorsal horn in the development of neuropathic pain (NP) in rats.Methods Forty-eight male Sprague-Dawley rats,weighing 200-250 g,in which intrathecal catheters were successfully implanted without complications,were randomly divided into 4 groups (n =12 each):sham operation group (group S),group NP,normal saline group (group NS) and minocycline group (group M).NP was induced by chronic constrictive injury of the sciatic nerve in NP,NS and M groups.Minocycline 50 μg was injected intrathecally once a day for 7 consecutive days starting from 1 day before ligation of the sciatic nerve in M group,while the equal volume of normal saline was given in stead of minocycline in NS group.Mechanical pain threshold and thermal pain threshold were measured at 1 day before ligation of the sciatic nerve (baseline,T0) and 1,3,7 days after ligation of the sciatic nerve (T1-3).Then the rats were sacrificed at T3 and the lumbar segment (L4,5) of spinal cord was removed for determination of the expression of CD46,CD55,CD59 protein and mRNA in the dorsal horn of spinal cord by Western-blot and RT-PCR,respectively.Results Compared with S group,mechanical pain threshold and thermal pain threshold were significantly decreased at T1-3 and the expression of CD46,CD55 and CD59 protein and RNA was down-regulated at T3 in NP,NS and M groups (P ＜ 0.05).Compared with groups NS and NP,mechanical pain threshold and thermal pain threshold were significantly increased at T2.3 and the expression of CD46,CD55 and CD59 protein and RNA was up-regulated at T3 in M group (P ＜ 0.05).Conclusion The down-regulated expression of membrane-bound complement regulatory proteins in spinal cord dorsal horn and abnormal activation of the complement are involved in the development of NP in rats.

Objective To evaluate the role of MCP/DAF expression in the spinal cord in the development of neuropathic pain (NP) induced by chronic constrictive injury (CCI) of the sciatic nerve in rats.Methods Twenty-four Sprague-Dawley rats transfected with MCP/DAF,weighing 200-250 g,were randomly divided into 2 groups (n =12 each) using a random number table:sham operation of transfected rat group (Rsham group) and CCI of transfected rat group (RCCI group).Twenty-four healthy male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 2 groups (n =12 each) using a random number table:sham operation of normal rat group (Nsham group) and CCI of normal rat group (NCCI group).The right sciatic nerve was exposed and 4 loose ligatures wen placed on the sciatic nerve at 1 mm intervals with 4-0 catgut in RCCI and NCCI groups.The right sciatic nerve was only exposed in Rsham and Nsham groups.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured at 1 day before operation (baseline) and 1,3 and 7 days after operation.The animals were sacrificed after measurement of pain threshold on 7 days after operation and the L4,5 segment of the spinal cord was removed for determination of the expression of OX-42 (by immuno-histochemistry) and MCP mRNA and DAF mRNA (by RT-PCR).Results Compared with Nsham group,the PWT and PWL were significantly decreased on 1,3 and 7 days after operation,the expression of OX-42 was up-regulated,and the expression of MCP mRNA and DAF mRNA was down-regulated in NCCI group (P ＜ 0.05),and no significant changes were found in the PWT and PWL on 1,3 and 7 days after operation and expression of OX-42(P ＞ 0.05),and the expression of MCP mRNA and DAF mRNA was up-regulated in Rsham and RCCI groups (P ＞ 0.05).Compared with NCCI group,the PWT and PWL were significantly increased on 1,3 and 7 days after operation,the expression of OX-42 was down-regulated,and the expression of MCP mRNA and DAF mRNA was up-regulated in RCCI group (P ＜ 0.05).Conclusion Up-regulation of MCP/ DAF expression in the spinal cord can inhibit the development of NP in rats and regulation of activation of microglias in the spinal cord is involved in the mechanism.