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Objective:To investigate the whole genome characteristics of coxsackievirus A4 (CVA4) circulating in Qingdao city.Methods:Four CVA4 isolates circulating in Qingdao city during 2013 to 2015 were selected. Whole genome sequences of these strains were amplified by one-step reverse transcription-polymerase chain reaction (RT-PCR). Sequence alignment and phylogenetic analysis were performed using MEGA7.0 software package. Genetic recombination analysis was performed using similarity plots 3.5.1 software package.Results:Phylogenetic analysis showed that based on the sequences of the whole genome and P1, P2 and P3 regions, HS312/QD/CHN/2013 and HS605/QD/CHN/2014 strains together with the early domestic isolates belonged to the same clade, while FY218/QD/CHN/2015 strain and CV-A4/P1033/2013/China strain collected in Wenzhou in 2013 formed another clade in each phylogenetic tree. HS144/QD/CHN/2014 strain belonged to the same clade as HS312/QD/CHN/2014, HS605/QD/CHN/2014 and the early domestic CVA4 isolates in the phylogenetic tree based on the P1 region, but formed a separate clade in the phylogenetic trees based on the whole genome, P2 region and P3 region. Genetic recombination analysis revealed that there was genetic recombination between HS144/QD/CHN/2014 strain and the CVA2 strain of CV-A2/P373/2013/China isolated in mainland China in 2013 in the region of 2C-3D (5 081-7 301); FY218/QD/CHN/2015 and CV-A4/P1033/2013/China strains were highly homologous and recombination signal sequences were detected in the region of 2A-2B (3 821-4 161) between the two strains and the CVA2 strain of CV-A2/P373/2013/China.Conclusions:The CVA4 isolates circulating in Qingdao city presented obvious genetic diversity at the genome-wide level.
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Objective@#To investigate the etiology spectrum of hand foot and mouth disease (HFMD) and to analyze the molecular characteristics of Coxsackievirus A10 and A6 in Qingdao in 2014.@*Methods@#Throat swabs of HFMD cases were tested for total enteroviruses (EVs), EV-A71, CV-A16, CV-A10 and CV-A6 by multiplex real time RT-PCR. Other EV serotypes were identified through the sequences of partial VP1 gene. The full-length of VP1 gene of CV-A10 and CV-A6 were amplified and sequenced. The sequences were phylogenetically analyzed using MEGA 5.0 software package.@*Results@#A total of 1727 HFMD patients were detected in 2014 and 11serotypes of enteroviruses were identified. EV-A71(38.0%, 410/1078), CV-A16(28.8%, 311/1078), CV-A10(14.1%, 152/1078)and CV-A6(3.2%, 34/1078)were the most dominant pathogen in 2014 in Qingdao. Proportions of CV-A10 in enterovirus infected children varied dramatically in different ages(χ2=15.19, P=0.001), showing a downtrend with age. Proportion of CV-A10 in inpatients was much higher than that in outpatients(χ2=49.1, P <0.001), while 60% of those inpatients showed nervous system damage symptoms, with pathological reflex or disappearance of physiological reflex. Phylogenetic analysis of complete VP1 sequences showed that all CV-A10 strains identied in this study belonged to genotype C, 71.7% of which located in branch C5; all CV-A6 strains belonged to genotype D while 83.3% of which located in branch D5.@*Conclusions@#EV-A71, CV-A16, CV-A10 and CV-A6 were the prevalent pathogens of HFMD in Qingdao in 2014. CV-A10 was more frequently detected in lower age group with HFMD, which can cause damage to nervous system. Strains of branch C5 in genotype C were the dominant strains of CV-A10 circulated in Qingdao in 2014 while strains of branch D5 in genotype D were the major strains of CV-A6.
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Objective@#To illuminate the gene characteristics and clinical characterization of Coxsackievirus B5 (CV-B5) strains isolated from patients with sevre hand, foot and mouth disease (HFMD) in Qingdao city.@*Methods@#A total of 1 844 patients of HFMD were consecutively admitted to Qingdao Women and Children's Hospital from 2013 to 2014. Information of the study population described above was collected retrospectively. The samples were collected from at least 1 site (throat swab, cerebrospinal fluid), which viral nucleic acid extracted and the entire VP1 gene sequences of CV-B5 isolates were amplified and sequenced, then the homology and phylogeny analysis were conducted by MEGA7.0. The prototype Faulkner strain and other VP1 amino acid sequences were derived from the GenBank database.@*Results@#A total of 8 CV-B5 positive cases were obtained, including 4 males and 4 females; 6 severe hospitalized cases and 2 outpatients. The age of 6 hospitalized patients ranged from 3 to 48 months, with a median of 26 months. For the six inpatients, fever, convulsions vomiting, diarrhea and rash were the main clinical manifestation, and all combined with viral encephalitis. Compared with the prototype strain Faulkner, in the VP1 region,the nucleotide and the amino acid homologies was 77.3%-78.8% and 95.5%-97.0% respectively. Five out of the six severe cases with substitution of serine (S) to asparagine (N) at amino acid site 95 in the VP1 region. The sequences of 8 CV-B5 strains were classified into genogroup D.@*Conclusion@#Hand, foot and mouth disease associated with CV-B5 virus infection can result in nervous system involvement and the main complication was viral encephalitis. The CV-B5 strains associated with severe hand, foot and mouth disease had high nucleotide homology and present a certain regional aggregation.
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Objective To investigate the molecular characteristics of coxsackievirus A12 ( CV-A12) and to understand the clinical manifestations of severe hand, foot and mouth disease (HFMD) caused by CV-A12 in Qingdao. Methods Throat swabs of HFMD, herpangina and influenza-like cases from 2011 to 2016 were detected for enteroviruses ( EVs) in Qingdao. Human rhabdomyosarcoma ( RD) and human la-ryngeal carcinoma (Hep-2) cells were used for virus proliferation and CV-A12 strains were identified through a semi-nest RT-PCR. The full-length of VP1 gene of CV-A12 strains was sequenced and phylogenetically an-alyzed using MEGA7. 0 software package. Clinical data of severe HFMD cases positive for CV-A12 were col-lected and analyzed. Results CV-A12-positive HFMD, herpangina and influenza-like cases accounted for 0. 3％(18/6798), 1. 2％(2/169) and 0. 1％(1/676) in Qingdao, respectively. Most of the HFMD caused by CV-A12 in children were mild before 2013 (84. 6％, 11/13), while hospitalized severe cases with neurological symptoms (100％, 5/5) became more common after 2013. Phylogenetic analysis of the VP1 region revealed that CV-A12 strains worldwide could be divided into two genotypes, A and B. All of the CV-A12 strains successfully sequenced in Qingdao from 2011 to 2016 belonged to genotype B, and 88. 9％(16/18) of them belonged to subgenotype B2. All hospitalized severe cases of CV-A12-caused HFMD after 2013 were associated with strains in branch B2b of subgenotype B2. Conclusion CV-A12 was one of the pathogens causing HFMD, herpangina and influenza-like illness in children in Qingdao. Strains of genotype B2 were the predominant CV-A12 strains circulating in Qingdao in recent years. CV-A12-caused HFMD might complicated by nervous system damage.
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Objectives: To assess the analgesic efficacy of transversus abdominis plane [TAP] block in patients undergoing colorectal surgery [CRS]
Materials and Methods: The databases of PubMed, ISI Web of Science, and Embase were searched, and randomized controlled studies [RCTs] that compared TAP block to control for relief of postoperative pain in patients who underwent CRS were included. Outcomes, including postoperative pain at rest and with movement, morphine use, postoperative nausea and vomiting, and the length of hospital stay, were analyzed using STATA software. The weighted mean differences [WMDs] with 95% confidence intervals [95% CIs] or relative risk with 95% CI were used to present the strength of associations
Results: A total of 7 RCTs with 511 patients were included. The results of this study suggested that TAP block significantly relieved postoperative pain during postanesthetic recovery after CRS at rest and during movement [WMDs were -0.98 [95% CI -1.57 to -0.38] and -0.68 [-1.07 to -0.30], respectively], and also decreased pain intensity during movement 24 h after CRS [WMD: -0.57 [95% CI -1.06 to -0.08]]. TAP block significantly reduced opioid consumption within 24 h when compared to controls, with a WMD of 15.66 [95% CI -23.93 to -7.39]. However, TAP block did not shorten the length of hospital stay
Conclusions: TAP block was an effective approach for relief of postoperative pain and reduced postoperative consumption of morphine. More RCTs with large sample sizes are required to confirm these findings
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Objective To investigate the etiology of hand,foot and mouth disease (HFMD) and to analyze the genetic characteristics of three prevalent enteroviruses in Qingdao in 2015. Methods City-wide surveillance data in 2015 were used to analyze the epidemiological characteristics of HFMD in Qingdao. RNA samples extracted from throat swab of HFMD cases were examined for general enteroviruses (EVs) such as enterovirus 71(EV71), coxsackievirus A16 (CA16) and CA6 by real-time RT-PCR. Full-length VP1 genes of EV-positive specimens were amplified and sequenced. Sequencing results were phylogenetically analyzed using MEGA 7.0 software package. Results A total of 804 patients with HFMD were identified from 1 176 patients in 2015. CA6 (41.4%),EV71 (31.6%) and CA16 (15.3%) were the predominant EVs causing HFMD. Children 5 years of age and under accounted for 80.3% of the 804 HFMD cases. CA6 was responsible for 48.9% of HFMD cases in children under 3 years old. The epidemic subtypes of EV71 and CA16 in Qingdao were C4a and B1b,respectively. Twenty-eight randomly selected CA6 strains were all classified into type A genome. Conclusion CA6, EV71 and CA16 were the prevalent pathogens causing HFMD in Qingdao in 2015. CA6 became the most predominant pathogen,mainly targeting children under 3 years old. C4a remained the prevailing subtype of EV71,while different from the co-prevalence of B1a and B1b subtypes in the past,B1b became the predominant subtype of CA16. CA6 strains circulating in Qingdao in 2015 mainly belonged to type A genome and evolved into multiple smaller branches. However, CA6 strains isolated in Qingdao in 2015 and 2013 located in different branches.
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Objective To investigate the etiology spectrum of hand , foot and mouth disease ( HFMD) and to analysis the molecular characteristics of three predominant human enterovirus stains in Qingdao in 2013.Methods The total enterovirus (EV) strains and strains of EV71, CVA16 and CVA6 in throat swabs of HFMD cases were detected by using multiplex real time RT-PCR.The full-length of the viral VP1 genes of the EV strains were amplified and sequenced .The sequences were phylogenetically analyzed by using the MEGA5.0 software package .Results A total of 841 patients with mild HFMD and 107 patients with serious HFMD were recruited in this study and 64 .3%of them were positive for EV .The predominant pathogens were EV71 (44.8%), CVA6 (28.2%) and CVA16 (9.5%) in 2013.CVA6 replaced CVA16 as the second most common pathogen for HFMD , accounting for 42.7% of all pathogens in children aged less than 3 years and 22.2%of all pathogens in the serious patients .The proportions of CVA6 in the etiology spectrum showed a downtrend along with the increasing age of the patients (P<0.001).Phylogenetic analy-sis of the complete VP1 gene sequences showed that all of the EV 71 strains identified in this study belonged to the subgenotype C4 (evolutionary branch C4a) and all of the CVA16 strains belonged to the subgenotype B1 (evolutionary branches B1a and B1b).There were 6 genogroups (A to F) regarding to the VP1 gene of CVA6 and all of the CVA6 strains identified in this study belonged to genogroups A and D .Among the CVA6 strains isolated in Qingdao in 2013, 83.9% belonged to genogroup A, while the rest 16.1% belonged to genogroup D.66.7%of the CVA6 strains isolated in 2012 belonged to genogroup A, while the rest 33.3%belonged to genogroup D .All of the CVA6 strains isolated from year 2008 to 2011 in Qingdao belonged to genogroup D.Conclusion EV71, CVA6 and CVA16 were the prevalent pathogens responsible for the de-velopment of HFMD in Qingdao in 2013.The proportions of CVA6 strains in the etiology spectrum showed a downtrend with the increasing age in children .C4a was the major subtype of EV71 strains circulating in Qingdao in 2013, while B1a and B1b were the major subtypes of CVA16 strains.The pattern of endemic cir-culation of CVA6 strains showed a trend of changing from genogroup D to A from year 2008 to 2013 .
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To study the anti-tumor metastatic constituents in Rhodiola wallichiana (HK) S H Fu var Cholaensis (Praeg) S H Fu, chemical constituents were isolated and purified by repeated column chromatography (silica gel, Toyopearl HW-40C and preparative HPLC). Their structures were elucidated on the basis of spectral data analysis. The anti-tumor metastasis assay was applied to evaluate the activities of the isolated compounds. Ten compounds (1-10) were isolated and their structures were identified by comparison of their spectral data with literature as follows: syringic acid (1), salidroside (2), tyrosol (3), scaphopetalone (4), berchemol (5), 2,6-dimethoxyacetophenone (6), rhobupcyanoside A (7), miyaginin (8), chavicol-4-O-β-D-apiofuranosyl-(1 --> 6)-O-β-D-glucopyranoside (9), eugenyol-O-β-D-apiofuranosyl-(1 --> 6)-O-β-D-glucopyranoside (10). Compounds 4-6 and 8-10, were isolated from this genus for the first time, while compound 7 was isolated from this plant for the first time. Compounds 2, 6-8 showed positive anti-tumor metastatic activities, and compounds 2 and 8 showed significant anti-tumor metastatic activities.
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Humanos , Antineoplásicos Fitogénicos , Farmacología , Línea Celular Tumoral , Metástasis de la Neoplasia , Rhodiola , QuímicaRESUMEN
Objective To study the whole-genome evolution of influenza B viruses prevalent in Qingdao from 2006 to 2011 .Methods RNA was extracted from influenza B viruses isolated in Qingdao from 2006 to 2011 .Each gene segment was amplified by reverse transcription polymerase chain reaction ( RT-PCR) and then sequenced .Gene sequences of each virus were determined and assembled by using Sequench -er software .A phylogenetic analysis for each gene segment was conducted by using MEGA 5.0 software pack-age.Results The phylogenetic tree of hemagglutinin ( HA) gene showed that 13 strains from 2006 to 2009 belonging to V1 clade of Victoria lineage were B/Malaysia/2506/2004-like viruses,and 12 strains from 2009 to 2011 belonging to the V2 clade of Victoria lineage were B/Brisbane/60/2008-like viruses.Moreover, strains of Yamagata lineage were all B/Florida/4/2006-like viruses including 5 strains of Y1 clade circulated from 2006 to 2008 and 7 strains of Y2 clade circulated from 2010 to 2011, respectively.The analysis of whole-genome evolution showed that 3 viruses of V2 clade presented 5+3 reassortment and 1 virus presented 1+7 reassortment.All reassortant strains matched with the vaccine strains of the present and previous season . The Yamagata and Victoria lineage strains belonged to genotype 2 and genotype 15,respectively.Compared with vaccine strains , the HA1 protein of Victoria lineage strains showed mutations at amino acid sites of H14Q, L58P, N129S, I146V, N171D and R279K, while R48K, K88R, P108A, N116K, S150I, N165Y, D196N,N202S and S229G amino acid mutations were mainly detected in Yamagata lineage strains .The sites 116 and 129,150,165,196 and 202 located in the 120,150,160 and 190 loops,respectively,which had been previously determined to be the hotspots under positive selection .Conclusion Both Yamagata and Victoria lineages of influenza B viruses were prevalent in Qingdao and evolved continuously from 2006 to 2011 .The selective pressure that a vaccine would provide was only to virus strains belonging to the same lineage ,sug-gesting a bivalent vaccine may be better for the induction of protective immunity .
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Objective To analyze the etiologic spectrum of hand, foot and mouth disease (HFMD) and the molecular characteristics of coxsackievirus A10 (CVA10) strains isolated in Qingdao from year 2010 to 2012.Methods Throat swab specimens were collected from patients with HFMD to detect to-tal enteroviruses ( EVs) , EV71 and CVA16 strains by multiplex real time RT-PCR.The EV-positive speci-mens were further detected by a semi-nested RT-PCR to amplify the sequence of viral genes encoding VP1. The serotypes of EVs were identified based on the sequences of genes encoding VP1.The full-length gene se-quences encoding VP1 of CVA10 isolates were amplified and sequenced. The phylogenetic analysis was con-ducted by using MEGA5.0 software package.Results A total of 1919 outpatients with mild HFMD and 1336inpatients with serious HFMD were recruited in this study .CVA16 strains were the predominant patho-gen for outpatients in year 2010 ( prevalence rate of 53%) and 2012 ( prevalence rate of 73%) .EV71 strains were the predominant pathogen for outpatients in year 2011 ( prevalence rate of 78%) and inpatients in year 2010 ( prevalence rate of 70%) and 2011 ( prevalence rate of 86%) . Some serotypes other than CVA16 or EV71 were the predominant pathogens for inpatients in year 2012 ( prevalence rate 44%) . CVA10 strains were identified in 12 patients with HFMD in year 2010and 17 patients with HFMD in 2012. The full-length gene sequences encoding VP1 of 23 CVA10 isolates were successfully amplified and se-quenced.The phylogenetic analysis showed that the 23 CVA10 isolates all belonged to genotype C and could be further divided into six clades.The genes encoding VP1 of the 23 CVA10 strains shared 97.3% to 1000.%homologies in amino acid sequences.The CVA10 isolates were also similar to their counterparts isolated from other regions during the same period.Conclusion CVA16 and EV71 strains were the preva-lent pathogens of HFMD in Qingdao from year 2010 to 2012, co-circulated with some other serotypes of EVs, especially CVA10 strains.The CVA10 strains belonged to genotype C and shared high homologies among them and their counterparts circulated in other regions during the same period.
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The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Electroacupuntura , Métodos , Esofagectomía , Dolor , Manejo del Dolor , Métodos , Complicaciones Posoperatorias , Terapéutica , Estudios ProspectivosRESUMEN
During the on-pump coronary artery bypass grafts surgery, ischemia/reperfusion injury would happen. Ischemia preconditioning could increase the tolerance against subsequent ischemia and reduce the ischemia/reperfusion injury. However the clinical outcomes of the available trials were different. We searched the Cochrane Central Register of Controlled Trials on The Cochrane Library [Issue 3, 2013], theMedline/PubMed and CNKI in March 2013. RevMan 5.1.6 and GRADEprofiler 3.6 were used for statistical analysis and evidence quality assessment. Heterogeneity was evaluated with significance set at P<0.10. Eighteen randomized controlled trials were included. There were no differences on in-hospital mortality, postoperative myocardial infarction morbidity between ischemia preconditioning and control groups. The heterogeneity of creatine kinase-MB level 24 hours after surgery was obvious. The differences of 72 hours area under the curve of cardiac troponinT [mean differences of -14.50, 95% confidence interval of -21.71 to -7.28] and troponin I [mean differences -181.79, 95% confidence interval of -270.07 to -93.52] after surgery were observed. All the 18 trails, the positive and the negative results were equal. The meta-analysis results should be interpreted with caution due to limited effective data. Because of high cost-effectiveness, ischemia preconditioning could not be denied completely. Large-scale randomized studies are needed, with the operation procedures and included criteria being more specific
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The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).
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<p><b>OBJECTIVE</b>To investigate the evolution features of whole-genome of influenza virus H3N2 prevalent in Qingdao from year 2007 to 2011.</p><p><b>METHODS</b>The RNA of 58 strains of influenza virus H3N2 prevalent in Qingdao between 2007 and 2011 was extracted and all segments amplified by RT-PCR. The sequence was then detected and assembled by software Sequencer. A total of 589 strains of influenza virus H3N2 with more than 300 amino acid recorded by GenBank were selected. The phylogeny and molecular features of all gene segments were analyzed by software Mega 5.0, referred by the heavy chain of hemagglutinin (HA1).</p><p><b>RESULTS</b>Hemagglutinin (HA) genes of influenza virus H3N2 prevalent in Qingdao between year 2007 and 2011 formed a single trunk of phylogenetic tree. Every prevalent strain originated in last season. The analysis of the evolution of whole genome found that reassortment virus strains were prevalent between year 2009 and 2010, but between 2010 and 2011 there were two series of prevalent strains, which showed complicated reassortment. Compared with the vaccine strains, the variant amino acids of protein of virus HA1 between year 2007 and 2011 were 8, 6, 6, 8 and 11, involving 13 antigenic sites. The sequence analysis of M2 protein showed that the isolated influenza virus H3N2 mutated in amino acid site 31, from serine to asparagine (S31N). HA1 gene of influenza virus H3N2 isolated in Qingdao between 2007 and 2011 shared the similar phylogenetic tree with the globally prevalent strain. The comparison of the sequence and the analysis of the antigenicity found co-infection between H3N2 and A/H1N1 in the strain A/Qingdao/F521/2011.</p><p><b>CONCLUSION</b>The evolution features of all segments of influenza virus H3N2 prevalent in Qingdao between year 2007 and 2011 were complicated.</p>
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Humanos , China , Evolución Molecular , Genoma Viral , Glicoproteínas Hemaglutininas del Virus de la Influenza , Genética , Subtipo H3N2 del Virus de la Influenza A , Genética , Filogenia , ARN Viral , Virus Reordenados , Genética , Análisis de Secuencia , Proteínas de la Matriz Viral , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To investigate characteristics of the whole-genome of influenza A H1N1 virus circulated in Qingdao from year 2009 to 2011.</p><p><b>METHODS</b>RNA of 35 influenza A H1N1 virus isolates circulated in Qingdao between year 2009 and 2011 was extracted and all segments were amplified by RT-PCR. The sequence was then detected and assembled by software Sequencher.25 HA full-length sequences published on GenBank were selected as reference. While MEGA 5.0 software package was explored for phylogenetic analysis to characterize the molecular feature with reference to the whole-genome sequence and the hemagglutinin (HA).1068 HA sequences of influenza A H1N1 virus isolated worldwide from August 2010 to March 2011 were downloaded for amino acid mutation analysis.</p><p><b>RESULTS</b>On the HA genes phylogenetic tree, the virus were separately divided into 4 clades in 2009-2010 and 2010-2011 surveillance season, each with a preponderant epidemic clade. The homogeneity of nucleotide and amino acids of HA isolates were 99.6%-99.9% and 99.1%-99.8% respectively in 2009-2010 surveillance season; 99.1%-99.6% and 98.2%-99.1% respectively in 2010-2011 surveillance season. The homogeneity of nucleotide and amino acids of the preponderant isolates were separately 98.8%-99.8% and 98.0%-99.6%. Compared with the vaccine strain, there were separately 14 and 12 variant amino acids of virus HA in the two surveillance season, involving 10 antigen sites and 5 positive selected sites. The sequence analysis of neuraminidase protein showed that the positions 247, 274 presented serine and histidine(S247, H274) respectively. The sequence analysis of M2 protein showed that the isolated A H1N1 viruses presented asparagine in amino acid site 31 (N31).</p><p><b>CONCLUSION</b>All the A H1N1 influenza virus circulated in Qingdao from year 2009 to 2011 presented continual variation and therefore caused antigenic drift. All the isolations were adamantane-resistance, but susceptible to inhibitors of neuraminidase.</p>
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Humanos , Secuencia de Aminoácidos , China , Epidemiología , Genoma Viral , Glicoproteínas Hemaglutininas del Virus de la Influenza , Genética , Subtipo H1N1 del Virus de la Influenza A , Clasificación , Genética , Gripe Humana , Epidemiología , Virología , Neuraminidasa , Genética , Filogenia , ARN Viral , Análisis de Secuencia de ProteínaRESUMEN
The study is to investigate the effect of angiotensin II (Ang II) and its receptor blockers on migration and endothelin-1 (ET-1) expression of rat vascular adventitial fibroblast subpopulations. Vascular adventitial fibroblasts were individually expanded by using cloning rings, and the effects of Ang II on the migration of adventitial fibroblast subpopulations were evaluated by Transwell. Fluorescence quantitative-PCR detected the expression of preproET-1 mRNA induced by Ang II, and its receptor antagonists losartan and PD-123319. The concentration of ET-1 was determined by ELISA. It showed that spindle shaped and epithelioid shaped cells were isolated by using cloning rings, named as spindle cells and round cells. RT-PCR showed that fibroblast subpopulations did not have leukocytes, endothelial cells and smooth muscle cells, namely pure cell lines. Compared with respective control cells, two subpopulations had transferring ability. Ang II significantly improved round cells migration in a concentration-dependent manner, and had no obvious influence on spindle cells migration. Ang II (1 x 10(-8) - 1 x 10(-6) mol x L(-1)) significantly increased the expression of preproET-1 mRNA in round cells (P < 0.01), and had no significant effect on the expression of preproET-1 mRNA in spindle cells. Losartan blocked the expression of preproET-1 mRNA induced by Ang II in round cells, and had no significant effect on the expression of preproET-1 mRNA in spindle cells. The effects of Ang II and ET-1 receptor inhibitors on the release of ET-1 were similar to the expression of preproET-1 mRNA. The results indicate that there are two cell subpopulations: round cells and spindle cells in rat vascular adventitial fibroblasts. Ang II significantly improved cells migration, and increased the expression of ET-1 in round cell subpopulation. It suggested that there may be different migratory mechanisms in two cell subpopulations, and the two subpopulations may play a different role in vascular remodeling and reparative process.
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Animales , Masculino , Ratas , Angiotensina II , Farmacología , Antagonistas de Receptores de Angiotensina , Farmacología , Movimiento Celular , Células Cultivadas , Endotelina-1 , Genética , Metabolismo , Fibroblastos , Biología Celular , Metabolismo , Imidazoles , Farmacología , Losartán , Farmacología , Piridinas , Farmacología , ARN Mensajero , Metabolismo , Ratas Sprague-Dawley , Vasoconstrictores , FarmacologíaRESUMEN
Objective: To construct a recombinant adeno-associated virus (AAV) vector harboring human hypoxia inducible factor-1α (HIF-1α) gene and to express it in primarily cultured neurons in the hippocampus. Methods: Human HIF-1α gene was inserted into AAV vector plasmid pSNAV 2.0 to generate a recombinant plasmid pSNAV-HIF-1α, then pSNAV-HIF-1α was transfected into BHK-21 cells and selected by G418. The construction of rAAV-HIF-1α was achieved by infecting G418-resistant BHK-21 cells with recombinant herpes simplex virus. The recombinant rAAV-HIF-1α was confirmed by polymerase chain reaction(PCR). The titer of rAAV-HIF-1α was detected by dot-blot with digoxin labelled cytomegalovirus probe. The purity of rAAV-HIF-1α was detected by SDS-PAGE. The expression of HIF-1α protein was determined by Western blot. Results: PCR demonstrated that human HIF-1α gene was successfully inserted into rAAV-HIF-1α. The Virus titer was 1 × 1012/ml and the virus purity exceeded 98%. Western nblot analysis demonstrated that rAAV-HIF1α infected hippocampal neurons expressed HIF-1α protein. Conclusion: rAAV-HIF-1α has been successfully constructed and it is capable of expressing HIF-1α in cultured hippocampal neurons.
RESUMEN
<p><b>OBJECTIVE</b>To observe the effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease (AD).</p><p><b>METHODS</b>Thirty-two male SD rats (250-300 g) were randomized into 4 groups (n=8), including the normal control group without any treatment, AD model group with right intracerebroventricular injection of 2 µl Aβ25-35 (10 mg/m1), sham-operated group with right intracerebroventricular injection of 2 µl normal saline, and AD+ rAAV-HIF-1α group with right intracerebroventricular injection of 10 µl rAAV-HIF-1a (1×10¹² v.g./m1) one week after Aβ25-35 injection. The rats were sacrificed to detect the expression of HIF-1α and apoptosis of hippocampal neurons 5 weeks after Aβ25-35 or saline injection.</p><p><b>RESULTS</b>Western blotting showed that the expression of HIF-1α was significantly higher in AD+rAAV-HIF-1α group (451.59±34.39) than in normal control group (229.05±41.28) and sham-operated group (216.29±37.08) (P<0.05) without significant difference between the latter two groups. The apoptotic ratio of the hippocampal neurons was significantly higher in AD model group ([19.49±2.59]%) than in normal control group ([5.41±0.75]%) and sham-operated group ([5.28±0.66]%) in (P<0.05), and intracerebroventricular injection of rAAV-HIF-1α resulted in a significant reduction of the apoptotic ratio in the AD rats ([12.07±2.06]%) (P<0.05).</p><p><b>CONCLUSION</b>Intracerebroventricular injection of rAAV-HIF-1α can inhibit hippocampal neuronal apoptosis in the rat model of AD.</p>