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This study was aimed to understand the current status of the antimicrobial resistance and molecular distribution of Campylobacter in various poultry in Jiaodong area,to provide a basis for effective prevention and control of the Campy-lobacter risk to poultry products and human health.Campylobacter was isolated and identified from 565 cloacal samples collect-ed in the Jiaodong area from August to October 2021 through conventional bacterial isolation and culture,mass spectrometry,microbroth dilution and multilocus sequence typing(MLST).The drug resistance and molecular typing of 131 representative strains(67 Campylobacter jejuni and 64 Campylobacter coli)were studied separately.Antimicrobial resistance analysis indica-ted that 131 isolates were highly resistant to ciprofloxacin,nalixic acid and tetracycline,with resistance rates of 96.21%,96.21%and 95.45%,respectively.Except for 2 strains,62 strains of C.coli were completely resistant to these three drugs(100%).A total of 65 strains of 131 strains were multidrug re-sistant,and the overall multidrug resistance rate was 49.62%,among which 11 strains(16.42%)of C.jejuni were resistance to 3-5 antibiotics,and 54 strains(84.38%)of C.coli were re-sistance to 3-6 antibiotics.Among the isolates from different poultry sources,waterfowl isolates were the most resistant,fol-lowed by broiler isolates.The MLST typing results revealed 72 alleles and 35 sequence types obtained from 67 strains of C.je-juni,and the distribution was relatively dispersed,without a dominant ST type and homologous complex.A total of 27 alleles and 19 sequence types were obtained from 64 strains of C.coli.Moreover,59.38%(38/64)strains were homologous complex CC-828,in which the ST-1586 sequence type was most frequent,followed by ST-825.ST-1586,ST-9944 and ST-3735 were the main sources of C.coli in broilers,and ST-825 and ST-1586 were the main sources of C.coli in waterfowl.Differences in C.jejuni and C.coli carriage were observed among poultry in the Jiaodong area.Carriage of the two bacteria was more common in laying hens than in broilers and waterfowl.C.jejuni from poultry in the Jiaodong area was highly resistant to ciprofloxacin,nalixic acid and tetracycline,but had good sensitivity to other drugs.C.coli was highly resistant to a variety of antibiotics,and multiple drug resistance was common.St-type dispersal of C.jejuni showed high genetic diversity.C.coli was cloned and transmitted mainly by ST-1586 in broiler chickens and waterfowl.Poultry carry C.jejuni,which can cause serious diseases in humans.Therefore,dynamic monitoring of Campylobacter from poultry should be strengthened.
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In order to reveal the molecular mechanism of blue labeled genic male sterility (BM-type GMS) and utilize the heterosis of BM-type GMS, we used the anthers of white-seed plants WS (sterile) and light blue seed plants WF (normal fertility) as experimental materials to analyze the differences in gene expression between them by transcriptome technology. And we also verified the genes expressed in anthocyanin synthesis in this study. Compared with WF, a total of 2352 differentially expressed genes were detected in WS. According to GO functional annotation, these genes could be divided into 3 categories and 43 subgroups. They are mainly involved in biosynthesis, phenylpropane metabolism, L-phenylalanine catabolism, membrane components, plasma membrane, cytoplasm, ATP binding, protein serine/threonine kinase activity, etc. KEGG pathway analysis showed that there were 159 genes enriched in the phenylpropanoid biosynthesis pathway, followed by the phenylalanine pathway, including 136 differentially expressed genes. Other genes are also involved a variety of amino acid metabolism, purine metabolism, pyrimidine metabolism and sugar metabolism pathway. Related to anthocyanin metabolism, several structural genes of key enzymes were differentially expressed, and most of them were up-regulated in WF, while only Flavanone 3-hydroxylase (F3H) and colorless anthocyanin dioxygenase (ANS) were down-regulated. Quantitative real-time PCR showed that the expression of 10 genes related to anthocyanin metabolism had the same trend as that in transcriptome sequencing data. Sequence homology analysis showed that the two selected transcription factors (DN48762c2g1 and DN25944c0g1) are clustered into the same cluster as the transcription factors regulating anthocyanin biosynthesis in maize, rice and Arabidopsis thaliana, which might be candidate genes for the blue aleurone layer of light blue seed plants in wheat. And fluorescence quantitative analysis showed that the expression level of DN48762c2g1 and DN25944c0g1 in WF was significantly higher than that in WS. In conclusion, the genes related to the anthocyanin biosynthesis pathway are not only related to the blue grain trait, but also may be involved in the anther abortion of BM-type GMS.
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Objective To establish an analysis method for simultaneous determination of 13 sedative substances and their metabolites in blood by liquid-liquid extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology and to apply the method to actual cases. Methods The samples were extracted with ethyl acetate after an internal standard was added. The extract was condensed until it was nearly dry and then its residues were dissolved with methanol, filtered through 0.22 μm filter and finally determined. The 13 sedative substances and their metabolites were separated through the C18 chromatographic column, then gradient elution was performed on them with methanol and 20 mmol/L ammonium formate (containing 0.1% formic acid) solution. After that, they were determined in the electrospray positive ion mode and quantified by internal standard method. Results The 13 sedative substances and their metabolites in blood showed good linearity in the range of 5-200 μg/L with correlation coefficients ranging from 0.990 3 to 0.999 8. The detection limits were 0.1-1.0 μg/L. Recovery rates of sedative substances were in the range of 71.2%-93.4% when solutions with concentrations of 10, 50 and 200 μg/L were added. The deviations of intra-day and inter-day relative standard deviations (RSD) were not more than 8.6%. Accuracies (bias) were within ±9.8%. Conclusion This method is rapid, simple, effective and sensitive, and can be applied to analysis of 13 sedative substances and their metabolites in blood in forensic toxicology.
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Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Toxicología Forense , Hipnóticos y Sedantes , Espectrometría de Masas en TándemRESUMEN
We established a multiplex direct PCR for rapid detection of E.coli,Salmonella,Staphylococcus aureus,Listeria and Yersinia enterocolitica bacteria.Multiplex direct PCR primers were designed according to gene sequences of phoA (E.coli),inv A (Salmonella),nuc (S.aureus),hl y (Listeria),and ail (Y.enterocolitica).After the multiplex direct PCR were established,the specificity and sensitivity of primers were detected.Then,multiplex direct PCR was applied to examine 60 swine product samples,the detection specificity,accuracy and positive predictive value were calculated compared with the gold standard culture method.Results showed that multiplex direct PCR primers could be used for specific detection of E.coli,Salmonella,S.aureus,Listeria and Y.enterocolitica,with the minimal detectable limit of 10,1,100,1 and 1 CFU,respectively.For the examination of 60 swine product samples using multiplex direct PCR,15 were positive for E.coli,6 positive for Salmonella,21 positive for S.aureus,20 positive for Listeria,and 35 positive for Y.enterocolitica,with all positive detection rates higher than that of culture.The total detection sensitivity was 100%,accuracy was 94%,and positive predictive value was 81.44%.Multiplex direct PCR could be used for specific and sensitive detection of common food-borne pathogens,and the testing time was shorten to be 3 hours because of saving time for template extraction.Multiplex direct PCR might serve the detection of food-borne pathogens in food safety risk monitoring much better.
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Objective To investigate the minimum dose of contrast medium when using spectral CT in cranial CTA. Methods Three groups of patients were required to undergo head CTA examination because of their condition,but different doses of contrast agents were injected.Among them,group 30 mL, group B, group 40mL, group C and group 50 mL were A. Results In group A,the lowest rate of excellent image (23.33%), B group, C group had higher rate of excellent image (93.33%, 96.67%), comparison of the data P<0.05; C group the highest rate of adverse reaction of contrast agent (P<0.05),data comparison P<0.05, A group, B group significantly adverse reactions of contrast agent there is no difference between the situation (P>0.05). Conclusion The use of 40 mL iodide contrast medium in the procedure of cranial CTA examination is effective and safe.
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Objective To investigate the minimum dose of contrast medium when using spectral CT in cranial CTA. Methods Three groups of patients were required to undergo head CTA examination because of their condition,but different doses of contrast agents were injected.Among them,group 30 mL, group B, group 40mL, group C and group 50 mL were A. Results In group A,the lowest rate of excellent image (23.33%), B group, C group had higher rate of excellent image (93.33%, 96.67%), comparison of the data P<0.05; C group the highest rate of adverse reaction of contrast agent (P<0.05),data comparison P<0.05, A group, B group significantly adverse reactions of contrast agent there is no difference between the situation (P>0.05). Conclusion The use of 40 mL iodide contrast medium in the procedure of cranial CTA examination is effective and safe.
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<p><b>OBJECTIVE</b>From May 2009-January 2010, a total of 3768 biosamples were tested for influenza A (H1N1) infection at Zhengzhou center for disease control and prevention, China. 1452 cases were laboratory confirmed H1N1 infection and 2316 were considered suspected victims. To evaluate the current protocol of influenza A (H1N1) based on the epidemic situations of Zhengzhou, relationships among features were explored and whether additional clinical characteristics should be part of H1N1 diagnosis protocols were determined.</p><p><b>METHODS</b>Both clinical and epidemiologic findings as well as statistical analyses were described in this article. Test for independence between features related to the disease diagnosis has been proposed. Furthermore, logistic regression was carried out to measure the association among features and latent class analysis was performed to identify additional crucial features in laboratory confirmed H1N1 by building various latent models with different combinatorial features.</p><p><b>RESULTS</b>The mean generation time for H1N1 was estimated as 3.59 +/- 1.41 days (range = 2.01-7.26). The estimated infection rate was 0.258 +/- 0.088 3, and reproduction number was 1.94 (95% CI = 1.12-3.18). Our results revealed that the six features, including molecular detections using three separate primer/probe sets, gender, age and temperature, are all associated with clinical diagnosis of H1N1, and that three separate primer/probe sets for laboratory confirmed H1N1, age and temperature are associated with each other.</p><p><b>CONCLUSION</b>Additional clinical features applied into the H1N1 diagnosis with current three primers/probe sets can increase the diagnostic efficiency.</p>
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Adolescente , Adulto , Anciano , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Distribución por Edad , China , Epidemiología , Diagnóstico Diferencial , Subtipo H1N1 del Virus de la Influenza A , Gripe Humana , Diagnóstico , Epidemiología , Modelos LogísticosRESUMEN
The nucleotide sequences of P gene from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The P gene is 1,655 nucleotides long with two overlapping open reading frames (ORFs). The first ORF is 1530 nucleotides long and would produce P protein of 509 amino acid residues. The second ORF is 534 nucleotides long and would produce C protein of 177 amino acid residues. The first ORF produces a second mRNA transcript of 897 nucleotides long with an extra G nucleotide at position 751. Translation from this mRNA would produce V protein of 298 amino acid residues. The nucleotide and deduced amino acid sequence were compared with the homologous region of other PPRV isolates. At the amino acid level, the "China/Tib/Gej/07-30" shares homology of 86.10%-97.3%, 84.3%-94.9%, and 82.9%-96.3% for P, C, and V proteins respectively. Several sequence motifs in the P genes were identified on the basis of conservation in the PPRVs and the morbilliviruses.
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Animales , Femenino , Secuencia de Aminoácidos , China , Enfermedades de las Cabras , Virología , Cabras , Datos de Secuencia Molecular , Peste de los Pequeños Rumiantes , Virología , Virus de la Peste de los Pequeños Rumiantes , Química , Genética , Metabolismo , Fosfoproteínas , Química , Genética , Metabolismo , Análisis de Secuencia , Homología de Secuencia de Aminoácido , Proteínas Virales , Química , Genética , MetabolismoRESUMEN
Background Researches demonstrated that the long-term application of glucocorticoids can induce cataract. However, its molecular mechanism is unclear. Objective Present study was to investigate the effects of dexamethasone on the regulation of nuclear factor kappa B( NF-κB)/ inhibitor kappa B alpha( IκBα) line on human lens epithelial cells (LECs) and the LECs apoptosis. Methods Human LECs line(HLE2B3) were cultured and passaged in DMEM containing 20% fetal bovine serum and treated by different concentrations of dexamethasone(0. 01,0. 1,1,10,100 μmol/L) for 24,36 and 48 hours respectively. The LECs cultured in free-serum DMEM without dexamethasone were as blank control group. The expressions of IκBo: in the LECs were examined by reverse transcription polymerase chain reaction ( RT-PCR) and Western blot, and the expressions of NF-κB neucleoprotein in LECs were detected by Western blot after exposure to dexamethasone. The apoptosis rate of LECs was determined by flow cytometer. Results Agarose gel electrophoresis showed that the amplified gene fragment was coincident to designed one. The expressing level of NF-κB neucleoprotein in LECs was significantly lowed with the increase of dexamethasone concentration ( F = 36. 077 , P = 0. 004 ) , and that of IkBo: was evidently ascended ( F = 35. 741 ,P = 0. 002). In the same concentration of dexamethasone group,the expression of NF-κB in LECs showed the considerable alteration in different duration after treated of dexamethasone with the lowest expressing level in 36 hours, and significant differences were found in the expressing level between 24 hours and 36 hours ( P = 0. 002) and between 24 hours and 48 hours (P = 0. 01). The differences of expression of IκBá in LECs appeared the same pattern to NF-κB neucleoprotein. Flow cytometry showed that the apoptosis rate of LECs was obviously enhanced after action of dexamethasone in a dose-dependent manner, showing a significant difference among different groups ( F = 73. 261, P = 0.001). Conclusion It is implied that dexamethasone results in the pathogenesis and development of glucocorticoid cataract by up-regulating the expression of IκBα in LECs and suppressing the activity of NF-κB and herein induce the apoptosis of LECs at concentration-and time-dependent manner. This might be one of cellular and biological mechanisms of glucocorticoid cataract formation.
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The nucleotide sequences of M and F genes from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The M gene was 1 483 nucleotides in length with a single open reading frame (ORF), encoding a protein of 335 amino acids. The F gene was 2411 nucleotides in length, encoding a protein of 546 amino acids. The resulting nucleotide sequence and the deduced amino acid sequences were compared with the homologous regions of other PPRV isolates. The nucleotide sequences of M and F genes of the "China/Tib/Gej/07-30" was 92.4%-97.7% and 85.5%-96.1% identical to other PPRV isolates, respectively, while a homology of 97.0%-98.2% and 94.3%-98.2% could be observed at the amino acids level respectively. Several sequence motifs in the M and F genes had been identified on the basis of conservation in the PPRVs and the morbilliviruses. The 3' untranslated region of M gene was 443 nucleotides in length with 82.4%-93.5% identical to other PPRV isolates. The 5' untranslated region of F gene was 634 nucleotides in length with 76.2%-91.7% identical to other PPRV isolates.
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Animales , Secuencia de Aminoácidos , Secuencia de Bases , Datos de Secuencia Molecular , Peste de los Pequeños Rumiantes , Virología , Virus de la Peste de los Pequeños Rumiantes , Química , Clasificación , Genética , Filogenia , Homología de Secuencia de Aminoácido , Ovinos , Enfermedades de las Ovejas , Virología , Tibet , Proteínas Virales de Fusión , Química , Genética , Proteínas de la Matriz Viral , Química , GenéticaRESUMEN
Sequence analysis of a new norovirus (NV) isolated from Lanzou city of China was performed based on partial sequence of RNA dependent RNA polymerase (RdRp) and complete capsid protein (VP1) gene. The isolated strain CHN02/LZ35666 shared high sequence homology with GII-4 NVs. Nucleotide homologies of RdRp region and encoded capsid protein region were 90.4% -- 98.6% and 89.8% -- 95.7% , respectively, while amino acid homology of capsid protein region was 94.4% -- 97.4%. The analysis of GDD motif in RdRp region indicated this GDD motif of Lanzhou strain differed from those of the GII-4 predominant epidemic strains. Lanzhou strain formed an independent branch in GII-4 cluster in the phylogenetic tree based on nucleotide sequence of RdRp region and amino acid sequence of capsid protein. Sequence alignment revealed a mutation at the fourth key site of the receptor-binding interface in the strains isolated after 2002 compared with those of previous strains suggesting a possible change of binding pattern to HBGAs receptors.
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Humanos , Secuencia de Aminoácidos , Secuencia de Bases , Proteínas de la Cápside , Genética , China , Gastroenteritis , Virología , Datos de Secuencia Molecular , Norovirus , Clasificación , Genética , Filogenia , ARN Polimerasa Dependiente del ARN , Genética , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
<p><b>OBJECTIVE</b>To compare the efficacy of bicyclol tablets on patients infected with hepatitis B virus between genotype B and C.</p><p><b>METHODS</b>70 patients with chronic viral hepatitis B were selected. The patients divide into two groups: HBV genotypes B (26 cases) and HBV genotypes C (other 44 cases). All patients received bicyclol tablets orally 150 mg daily (50mg, tid, po) for 24 weeks. The efficacy were observed after 12 weeks and 24 weeks.</p><p><b>RESULTS</b>After treatment for 24 weeks, the serum aminotransferase were decreased obviously, and HBV DNA levels turn to be negative with 19.2 percent (genotype B group) and 15.9 percent (genotype C group), respectively. The difference was not statistically significant between HBV genotype B and C.</p><p><b>CONCLUSION</b>Bicyclol not only has hepatoprotective activity but also inhibited virus replication in patients infected with HBV. The difference of the response to bicyclol therapy between HBV genotypes B and C was not statistically significant.</p>
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Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Compuestos de Bifenilo , Usos Terapéuticos , Genotipo , Virus de la Hepatitis B , Clasificación , Hepatitis B Crónica , Quimioterapia , VirologíaRESUMEN
<p><b>BACKGROUND</b>To study the arboviruses carried by mosquitoes collected in Hebei Province.</p><p><b>METHODS</b>Samples were collected from mosquito active sites and stored in liquid nitrogen till use. Pools of 20 to 30 mosquitoes were ground after sterilization, centrifugal supernant was inoculated onto C6/36 cell, cytopathic effect was observed for three sequential passages. Positive isolates were identified by IFA and RT-PCR.</p><p><b>RESULTS</b>Totally 1310 mosquitoes were collected from two villages of She county, Hebei province. They were divided into 46 pools and ground respectively. Thirteen positive isolates were obtained. Two isolates reacted with alphaviral antibodies and were amplified by alphaviral primers, nucleotide sequence showed the highest homology (98%) to Getah virus (AY702913.1), so the two isolates were identified as Getah virus.</p><p><b>CONCLUSION</b>Getah virus was isolated from mosquitoes in Hebei Province. This is the first report of isolating Getah virus from inland of China.</p>
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Animales , Arbovirus , Clasificación , Genética , Línea Celular , Análisis por Conglomerados , Culicidae , Virología , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADNRESUMEN
<p><b>OBJECTIVE</b>To analyse sonographical features of epididymis tuberculosis with caseous necrosis and improve the accuracy of ultrasonic diagnosis.</p><p><b>METHODS</b>Ultrasonic features of 12 cases with epididymis tuberculosis with caseous necrosis, confirmed by surgical pathology, was retrospectively analysed: size, location, echogenicity, degree of blood flow in the lesion.</p><p><b>RESULTS</b>In all 12 patients, the number of purified protein derivative test (PPD) presenting the positive reached to 67%, while cases with the pulmonary tuberculosis was 42%. The epididymal tuberculosis with caseous necrosis could present images of the whole or focal epididymal enlargement and abnormal shape. The lesions internal echoes were irregular, inhomogeneous increased isoechoic echogenicity or hypoechoic, including bad-defined, irregular, homogeneous hypoechoic or anechoic. In lesional location, 4 out of 12 patients (33%) possessed richly color blood flow in Color Doppler Power Imaging(CDPI). The testicular enlargement with increased internal echoes and the testicular hydroceles were partly (58%, 7/12) observed by ultrasound. There was no varicocele, scrotal wall thickening and inguinal lymph node enlargement.</p><p><b>CONCLUSION</b>The High Frequency Color Doppler images is helpful for the diagnosis and differential diagnosis of epididymis tuberculosis with caseous necrosis.</p>
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Adulto , Humanos , Masculino , Persona de Mediana Edad , Diagnóstico Diferencial , Epididimitis , Diagnóstico por Imagen , Patología , Necrosis , Estudios Retrospectivos , Tuberculosis de los Genitales Masculinos , Diagnóstico por Imagen , Patología , Ultrasonografía Doppler en ColorRESUMEN
A series of mutual prodrugs derived from gabapentin, pregabalin, memantine, venlafaxine were synthesized and their pharmacological properties to treat neuropathic pain were investigated in a rat model of chronic sciatic nerve constriction injury (CCI). In vivo evaluation demonstrated that the mutual prodrugs 2002413A, 2002823A composed of two gabapentins, 2002414 composed of gabapentin and pregabalin were effective in reversal tactile allodynia in CCI rats. The prodrugs 2002413A, 2002414 had no significant influence on the rotarod activity. The result suggest that the prodrugs may be possible candidates for further development.