192 samples of Masked Palm Civet (Paguma Larvata) from Guangdong Province were inoculated in Vero-E6 cells. One sample which came from masked palm Civet didn't cause cytopathic effects (CPE) until fourth-passage on Vero-E6 cells. Infected cells emerged granulating, shrinking, rounding and falling off. After three times freeze-thaw, cells and culture medium were harvested for electron microscopy. Virus particles were nonenveloped, double capsid and icosahedral symmetry. This virus was designated Masked Palm Civet/China/2004 (MPC/04). Hemagglutination test indicated that the virus could agglutinate healthy human type O red cells, but not the red cells of SPF chicken, experimental common bovine, rat and guinea pig. This virus was tolerant to chloroform treatment, pH 3.0 and water bath 50 degrees C 1 h. 1 M MgCl2 treatment could enhance resistance of virus to heat and increase infectivity. In order to classify the strain on the molecular level, specific primers according to mammalian reovirus were used for Reverse Transcription Polymerase Chain Reaction (RT-PCR). Appropriate specific products were amplified by RT-PCR. NCBI BLAST analysis indicated that this segment shared the highest identity to mammalian reovirus serotype 1 (T1L) virus. So we can deduce this virus is a member of the Reoviridae.
Animals ; Base Sequence ; Cats ; virology ; Cattle ; Humans ; Molecular Sequence Data ; Phylogeny ; Reoviridae ; classification ; isolation & purification

Membrane (M) protein genes of 20 infectious bronchitis virus (IBV) strains isolated in China between 1995 and 2004 were sequenced and analyzed. The M genes of twenty isolates were composed of 672 to 681 nucleotides, encoding polypeptides of 223 to 226 amino acid residues. Variations of the deduced amino acids of M gene mainly occurred at positions 2 to 17 and 221 to 233, comparing with that of the IBV strain LX4. There were deletions or insertions in the M gene of Chinese isolates at amino acid position 2 to 6, leading to the loss or gain of a glycosylation site. Phylogenetic tree based on amino acid sequences of M genes from 20 Chinese isolates and 34 reference strains showed that they were classified into five distinct clusters. Most of the Chinese IBV strains were included in clusters II and IV, forming distinct groups. The isolates in cluster II showed a close evolutionary relationship with Taiwan isolates. Furthermore, recombination especially the recombination between field isolates and vaccine strains had been observed while comparing the phylogeny of M genes with those of S1 and N genes.
Amino Acid Sequence ; China ; Genetic Variation ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Sequence Homology, Amino Acid ; Viral Matrix Proteins ; genetics

OBJECTIVETo investigate the effects of intrathecal ouabain and tizanidine injection for treatment of neuropathic pain in rats.

METHODSMale SD rats weighing 250-300 g were randomly divided into 5 groups (n = 6), namely the control group, ouabain group, tizanidine group, combined ouabain and tizanidine injection group, and the antagonist group. Intrathecal catheter was implanted 7 days before spinal nerve ligation to establish the neuropathic pain model. Mechanical withdrawal threshold (MWT) before and after intrathecal administration of the agents was recorded in the rats. Isobolographic analysis was performed to evaluate the interactions between the agents.

RESULTSIntrathecal injection of ouabain (0.25-5 microg) or tizanidine (0.5-5 microg) alone produced dose-dependent analgesic effect against the neuropathic pain (P < 0.05). Isobolographic analysis revealed a synergistic interaction between ouabain and tizanidine. Intrathecal pretreatment with atropine (5 microg) or yohimbine (20 microg) antagonized the effects of ouabain and tizanidine administered alone or in combination (P < 0.05).

CONCLUSIONIntathecal injection of ouabain or tizanidine produces dose-dependent analgesic effects against neuropathic pain, and their synergistic effect after combined injection probably involves the cholinergic transmission and alpha2 receptor.

Analgesics ; administration & dosage ; Animals ; Clonidine ; administration & dosage ; analogs & derivatives ; Injections, Spinal ; Ouabain ; administration & dosage ; Pain ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Nerves ; injuries

Objective To investigate the relationship between single nucleotide polymorphisms of tumor suppressor gene PTEN and nasopharyngeal carcinoma in Jiamusi Han population. Methods The blood samples of 132 patients with naso-pharyngeal carcinoma(nasopharyngeal carcinoma group)and 73 healthy people(control group)were selected from September 2008 to January 2018 in the First Affiliated Hospital of Jiamusi University. The whole genome DNA was extracted,and the pol-ymorphisms of rs532678 and rs701848 were detected by restriction fragment length polymorphism analysis. The relationship be-tween the polymorphism of PTEN gene and nasopharyngeal carcinoma was analyzed. Results The genotype and allele frequen-cy distributions of rs532678 and rs701848 loci were in line with the Hardy-Weinberg genetic balance law in the two groups (P > 0. 05). The genotypic frequency of CC,CT and TT at the rs532678 locus of PTEN gene in the control group was 0. 630, 0. 342 and 0. 027 respectively;and the allele frequency of C and T was 0. 801 and 0. 198 respectively. The genotypic frequency of CC,CT and TT at the rs532678 locus of PTEN gene in the nasopharyngeal carcinoma group was 0. 716,0. 265 and 0. 015 re-spectively;and the allele frequency of C and T was 0. 852 and 0. 147 respectively. There was no significant difference in geno-type distribution and allele frequency distribution at the rs532678 locus of PTEN gene between the two groups(P > 0. 05). The genotypic frequency of CC,CT and TT at the rs701848 locus of the PTEN gene in the control group was 0. 657,0. 342 and 0. 000 respectively;and the allele frequency of C and T was 0. 828 and 0. 171 respectively. The genotypic frequency of CC,CT and TT at the rs701848 locus of PTEN gene in the nasopharyngeal carcinoma group was 0. 424,0. 500 and 0. 075 respectively;and the allele frequency of C and T was 0. 674 and 0. 325 respectively. The frequencies of CT,TT genotype and T allele of rs701848 locus in the nasopharyngeal carcinoma group were significantly higher than those in the control group(P < 0. 05). The frequencies of CC genotype and C allele in the nasopharyngeal carcinoma group were significantly lower than those in the control group(P < 0. 05). The individual with CT + TT genotype at the rs701848 locus of PTEN gene had higher risk for naso-pharyngeal carcinoma(P < 0. 05,OR = 2. 606,95％ confidence interval:1. 439 - 4. 720). The risk for nasopharyngeal carcino-ma in the individual with CT + TT genotype was 2. 606 times as much as the individual carrying CC genotype. Conclusion The rs532678 polymorphism of PTEN gene is not associated with the susceptibility to nasopharyngeal carcinoma. The polymor-phism of rs701848 locus is associated with the susceptibility to nasopharyngeal carcinoma. The individual carrying CT + TT genotype has higher risk for nasopharyngeal carcinoma.

The entire S1 protein gene of five infectious bronchitis (IB) vaccine strains (JAAS, IBN, Jilin, J9, H120) used in China were compared with that of the IB field isolate CK/CH/LDL/97 I present in China. The nucleotide and deduced amino acid similarities between the five IB vaccine strains and the field strain, CK/CH/LDL/97 I, were not more than 76.4% and 78.7%, respectively. Phylogenetic analysis based on the S1 gene showed that the vaccine strains and the field strain belonged to different clusters and had larger evolutionary distances, indicating that they were of different genotypes. The five vaccine strains were used for protection test against challenge of the field isolate CK/CH/LDL/97 I. The chickens inoculated with five vaccine strains showed morbidity as high as 30%-100% after challenged with the CK/CH/ LDL/97 I strain. The organ samples at 5 days post challenge showed that the viral detection rates were 50%-90% and 10%-30% for trachea and kidney, respectively. The live attenuated vaccines only provided partial protection to the vaccinated chickens against heterologous IBV infection, CK/CH/LDL/97 I.
Animals ; Antibodies, Viral ; blood ; Chickens ; virology ; Coronavirus Infections ; prevention & control ; veterinary ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Membrane Glycoproteins ; genetics ; Phylogeny ; Poultry Diseases ; prevention & control ; Spike Glycoprotein, Coronavirus ; Vaccines, Attenuated ; immunology ; Viral Envelope Proteins ; genetics ; Viral Vaccines ; immunology

Artemisia annua also known as Qinghao, is a kind of traditional Chinese medicine. Its active ingredient is artemisinin, a sesquiterpene lactone compound with a peroxy bridging group structure. A. annua is an effective antimalarial drug. Artemisinin, a secondary metabolite in A. annua, can be induced by many physical and chemical factors, such as salinity, moisture, light, and plant hormones. Temperature, as an important growth factor, also has a great influence on the synthesis of artemisinin. This article aims to study the effect of high temperature on inducing artemisinin biosynthesis in A. annua. The A. annua seedlings were placed at 25, 40 °C, and the samples were taken after 0, 3, 12 and 36 h. The content of artemisinin in each sample was determined by liquid chromatography-mass spectrometry. Total RNA was extracted from the samples, and then transcriptome sequencing and real-time fluorescence quantitative PCR were used to quantitatively analyze the expression of the key enzyme genes in artemisinin synthesis pathway and competition pathway. The results showed that artemisinin content was increased by 20%, 42% and 68% after 3, 12, 36 h of treatment at 40 °C. The expression levels of FDS, ALDH1, CYP71AV1 and ADS were up-regulated by 4.3, 3.3, 2.5, 1.9 times, and the expression levels of SQS and BPS were down-regulated by 37% and 90% respectively. In summary, high temperature can promote the biosynthesis of artemisinin by promoting the expression of synthetase genes in artemisinin synthesis pathway and inhibiting the expression of synthetase genes in artemisinin-competition pathway.
Antimalarials ; metabolism ; Artemisia annua ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Plants, Medicinal ; metabolism ; Temperature