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<p><b>OBJECTIVE</b>To analyze the genetic characterization of the complete genome from a human coxsackievirus B3 strain A103/KM/09 isolated in Yunnan province, 2009.</p><p><b>METHODS</b>By using RT-PCR, all the eight fragments which containing about 1000 nucleotides and covering full viral genome, were sequenced. By using Mega 5.05,Geneious, RDP 3 and SimPlot 3.5.1 software, sequences were aligned with other enterovirus reference sequences. Phylogenetic and recombination analysis were also carried out.</p><p><b>RESULTS</b>The A103/KM/09 isolate genome showed 7389 nucleotides in length , encoding for 2185 amino acids. In the complete genome, the homology of nucleotide and amino acid among the seven coxsackievirus B3 isolates were 81.0%-88.0% and 95.7%-98.0%, respectively. There appeared 81.0% and 95.7% homology when compared with that of Nancy prototype strain. Results from the Phylogenetic analysis showed that the coxsackievirus B3 formed five distinct clades, I-V. Nucleotide divergence rates between clades were 16.2%-24.3% . The A103/KM/09 strain belonged to clade V. Clade V was further divided into four sub-clades,A-D. The nucleotide divergence between sub-clades was 4.3%-11.4%. Putative recombinant event for A103/ KM/09 was detected.</p><p><b>CONCLUSION</b>All coxsackievirus B3 isolates could be divided into five clades, with A103/KM/09 strain belonged to Clade V-D. Evolution of coxsackievirus B3 had occurred in China.</p>
Subject(s)
Child, Preschool , Humans , Male , Base Sequence , China , Epidemiology , Encephalitis, Viral , Epidemiology , Virology , Enterovirus B, Human , Genetics , Enterovirus Infections , Epidemiology , Virology , Genome, Viral , Phylogeny , Viral Proteins , GeneticsABSTRACT
To characterize the complete genome sequence of coxsackievirus B1 (CVB1) MSH/KM9/2009 strain isolated from Yunnan, China,2009. Eight overlapping clones covering the whole viral genome (excluding the poly-A tail) were obtained by RT-PCR and sequenced, and their nucleotide and amino acid sequences were compared with other known CVB1 strains. The genome of the CVB1 MSH/KM9/2009 strain had 7384 nucleotides in length, and contained a 741nt non-translated region (NTR) at the 5' end and a 94nt NTR at the 3' end. The entire open reading frame contained 6 549 nt, encoding a 2 183-aa polyprotein. In the coding region, there was no nucleotide deletion or insertion, but some changes of amino acid were unique. The complete genome sequence alignments showed that the CVB1 isolate MSH/KM9/2009 strain shared the highest nucleotide (80.9%, 81.6%, 80.5% and 80%) and amino acid (95.6%, 95.8%, 96.2% and 95.6%) identities to the CVB1 M16560, pmMC, Tucson B1 and CVB1Nm strain, respectively. Phylogenetic tree analysis showed that the MSH/KM9/2009, CVB1 M16560, pmMC, Tucson B1 and CVB1Nm strain clustered into same group. The newly isolated CVB1 strain MSH/KM9/2009 from Yunnan Province belonged to genotype CVB1.
Subject(s)
Child, Preschool , Female , Humans , Infant , Male , Base Sequence , China , Coxsackievirus Infections , Virology , Enterovirus , Classification , Genetics , Genome, Viral , Genotype , Open Reading Frames , Phylogeny , Viral Proteins , GeneticsABSTRACT
Objective To analyze the genetic characterization of the complete genome from a human echovirus 6 (Echo6) strain KM57-09 isolated in Yunnan,China,in 2009.Methods Using the RT-PCR,eight fragments containing about 1000 nucleotides which covered the whole viral genome were sequenced.The sequences were aligned with other reference cnterovirus sequences downloaded from the GenBank,using Mega 5.05,RDP 3 and SimPlot 3.5.1 softwares.Results Similar to the other human cntcrovirus,KM57-09 isolate genome appeared to have 7419 nucleotides in length,encoding for 2191 amino acids.In the complete genome,the rates of homology on nucleotide and amino acid among the seven Echo6 isolates were 79.3％-80.2％ and 93.3％ 94.4％,respectively as well as 79.3％ and 93.6％ of the rates of homology when compared with that of D' Amor prototype strain.In different segment of genome.The 2C 3A genome region was most similar to the HN-2-E25 strain,the 5' UTR,VP4,3D and 3' UTR genomc region were most similar to the CoxB5-Henan2010.In the VP1 gene,the rates of homology on nuclcotide and amino acid among the China isolates were 80.0％-96.0％ and 95.8％-99.0％,respectively,and showed 77.6％-96.0％ and 95.2％-99.0％ of the rates on homology when compared to the other Echo6 reference strains isolated from other countries or areas,respectively.Results from phylogenetic analysis showed that the Echo6 formed five distinct groups,A-E.The KM57-09 strain belonged to clade E.The nucleotide divergence between clades was 15.6％-23.3％.The putative recombinant event for KM57-09 was detected with RDP 3,SimPlot 3.5.1 and 3D sequence phylogenetic analysis.Conclusion All the Echo6 isolates could be divided into five clades,the KM57-09 strain belonged to Clade E.The Echo6 strains isolated in China were contributed to several different chains of transmission.
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To investigate the spatiotemporal properties of epileptiform activity in vitro, 400 microm-thick transverse hippocampal slices were prepared from juvenile rat and planar multi-electrode array (MEA) containing 60 electrodes was used to record the electrical activity induced by bath application of high potassium artificial cerebrospinal fluid (ACSF) on slices. Following successful induction of epileptiform bursts, phenobarbital sodium was applied to test for its inhibitory effects on bursting activity in different regions of slice. Region-specific characteristics of epileptiform activity and anticonvulsant actions of phenobarbital sodium in the hippocampal network were determined by comparing the population activity obtained from MEA. The results showed that: (1) 15 min after high-K+ ACSF application, rhythmic and synchronous epileptiform bursts could be detected from all CA sub-regions. Quantitative analysis indicates that the firing patterns of different CA sub-regions were not statistically different (P>0.05). However, no bursting activity was recorded from granular cells in dentate gyrus, only sparse spikes were observed, with frequency significantly lower than that in CA regions (P<0.05). (2) The high-K+-induced bursting activity could last for more than 40 min with stable bursting activities. (3) Bath application of 60 micromol/L phenobarbital sodium inhibited the bursting activities on hippocampal slice. Bursting activities in CA3c and CA1 were firstly suppressed. 10 min after the phenobarbital sodium application, strong bursting activities persisted only in some of pyramidal cells in CA3a and CA3b. These results show that MEA could be applied for studying the spatial and temporal properties of epileptiform activity in vitro, as well as the region-specific effects of anti-epileptic drugs.
Subject(s)
Animals , Male , Rats , Action Potentials , Physiology , Electrodes , Electroencephalography , Electrophysiological Phenomena , Physiology , Epilepsy , Hippocampus , In Vitro Techniques , Rats, Sprague-Dawley , Signal Processing, Computer-AssistedABSTRACT
<p><b>OBJECTIVE</b>To report the experience of surgical resection of Bismuth-Corlette type I and II hilar cholangiocarcinoma.</p><p><b>METHODS</b>From January 1998 and January 2008, 52 cases of Bismuth-Corlette type I and II hilar cholangiocarcinoma were operated on. The clinical data and long-term outcome of the patients was retrospectively analyzed.</p><p><b>RESULTS</b>Of the 52 cases, 44 cases (84.6%) received operation, 28 patients underwent radical resection (63.6%) and 16 patients (36.4%) underwent palliative resection.Seven patients were resected on caudate lobe and other section and lobe of the liver; among them, 2 patients received combined portal vein resection and 4 underwent combined hepatic artery resection respectively. Eleven cases developed postoperative complications and another one died in hospital. The median survival was 33.2 months in radical resection group, and 1-, 3-, 5-year survival rate was 82.6%, 47.8%, 34.7%, respectively, which was significant greater than those in the palliative resection group (41.6%, 16.6%, 8.3%, respectively) (P < 0.05). The median survival was 16.7 months in the palliative resection group.</p><p><b>CONCLUSIONS</b>The radical resection is still the best treatment for Bismuth-Corlette type I and II hilar cholangiocarcinoma. Intraoperative pathology for resection margin, and combined liver resection, portal vein resection and hepatic artery resection can help improve the radical resection rate.</p>
Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bile Duct Neoplasms , General Surgery , Bile Ducts, Intrahepatic , Cholangiocarcinoma , General Surgery , Follow-Up Studies , Hepatectomy , Hepatic Artery , General Surgery , Portal Vein , General Surgery , Prognosis , Retrospective Studies , Survival RateABSTRACT
Objective To explore the changes of nitric oxide(NO) and superoxide dismutase(SOD) in the serun and intestine mucosal and the treatment of ulcerative colitis(UC) with tanshinon in rats,Methods 30 mg of trinitro-benzene-sulfonic acid (TNBS) dissolved in 0.85ml of 50% ethanol was administrated intrarectally in Sprague-Dawley female rats to induce experimental colitis.After 7 days,the rats were divided into normal control, 0.9% saline and treatment group tanshinon 2ml?kg~(-1)?d~(-1) were intravenously.The therapeutic effect was evaluated by measuring the ponderal index,the surface area of the ulcers,macroscopical and histological score,activity of NO and SOD were measured in colonic tissue and serum all rats.Results Compared with the saline group,the ponderal index,the surface area of the ulcers,macroscopical and histological score,activity of NO level in the serum and intes- tine mucosal was decreased and the SOD increased of significantly in the treatment group(P