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1.
Chinese Journal of Analytical Chemistry ; (12): 121-129,中插38-中插43, 2024.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-1017636

RESUMEN

A rapid analytical method for simultaneous determination of 32 kinds of multi-residue veterinary drugs in eggs was developed using a modified QuEChERS technique based on a reduced graphene oxide-coated melamine sponge(r-GO@MeS)by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).The influences of graphene oxide(GO)concentrations,sponge dosages,and purification modes on drug recoveries were investigated during the purification process.The optimal purification conditions involved using a GO concentration of 0.5 mg/mL,a sponge dosage of 6.0 cm3/mL,and a dynamic purification mode of 5 extrusion cycles.Separation was achieved using an Agilent Eclipse Plus C18 RRHD column(100 mm×2.1 mm,1.8 μm),and quantitative analysis was performed by the external standard method using an electrospray ionization source(ESI)in multiple reaction monitoring(MRM)mode.The results showed that all 32 kinds of veterinary drugs exhibited good linear correlation with coefficients greater than 0.999,and matrix effects(MEs)ranging from?7.8%to 18.9%.The limits of detection(LODs)and quantification(LOQs)ranged from 0.2 to 10.2 μg/kg and from 0.6 to 28.0 μg/kg,respectively.The recoveries for the three spiked levels were in the range of 66.5%?117.5%,with intra-day and inter-day precision(Relative standard deviation)below 13.3%and 16.3%,respectively.The synthetic r-GO@MeS exhibited efficient matrix purification without the need of high-speed centrifugation or strong magnetic field assistance.This significantly shorted the sample pretreatment time and improved the convenience of the matrix purification process.Combined with UPLC-MS/MS,the method was suitable for the rapid determination of multi-residue veterinary drugs in eggs.

2.
J Hazard Mater ; 452: 131208, 2023 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-36966625

RESUMEN

Antibiotics are increasingly used and released into the marine environment due to the rapid development of mariculture, resulting in spread of antibiotic resistance. The pollution, distribution, and characteristics of antibiotics, antibiotic resistance genes (ARGs) and microbiomes have been investigated in this study. Results showed that 20 antibiotics were detected in Chinese coastal environment, with predominance of erythromycin-H2O, enrofloxacin and oxytetracycline. In coastal mariculture sites, antibiotic concentrations were significantly higher than in control sites, and more types of antibiotics were detected in the South than in the North of China. Residues of enrofloxacin, ciprofloxacin and sulfadiazine posed high resistance selection risks. ß-Lactam, multi-drug and tetracycline resistance genes were frequently detected with significantly higher abundance in the mariculture sites. Of the 262 detected ARGs, 10, 26, and 19 were ranked as high-risk, current-risk, future-risk, respectively. The main bacterial phyla were Proteobacteria and Bacteroidetes, of which 25 genera were zoonotic pathogens, with Arcobacter and Vibrio in particular ranking in the top10. Opportunistic pathogens were more widely distributed in the northern mariculture sites. Phyla of Proteobacteria and Bacteroidetes were the potential hosts of high-risk ARGs, while the conditional pathogens were associated with future-risk ARGs, indicating a potential threat to human health.


Asunto(s)
Antibacterianos , Microbiota , Humanos , Antibacterianos/farmacología , Genes Bacterianos , Enrofloxacina , Bacterias/genética , Bacteroidetes , Proteobacteria/genética
3.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-995264

RESUMEN

Objective:To investigate the role of dendritic cells (DC) in Chlamydia muridarum ( Cm) respiratory infection and their effect on adaptive immune response. Methods:C57BL/6 mice were exposed to 1×10 3 inclusion-forming units (IFU) of Cm through inhalation to establish the mouse model of Cm respiratory infection. The proportion of CD11c + MHCⅡ + DC and the expression of costimulatory molecules (CD40, CD80 and CD86) in spleen tissues were detected by flow cytometry on 0, 3 and 7 d after infection. The expression of IL-12p40, IL-10 and IL-6 at mRNA level in spleen tissues was detected by qPCR. Mouse splenic DC isolated on 7 d after Cm infection were sorted by magnetic beads and then transferred to recipient mice. Th1 response in the recipient mice was measured using intracellular cytokine staining 14 d after infection. Results:Cm respiratory infection induced massive infiltration of DC and promoted the expression of costimulatory molecules on splenic DC. The expression of IL-12 and IL-10 at mRNA level in splenic DC reached the peak on 3 d after infection. Transferring the splenic DC of Cm-infected mice into the recipient mice could alleviate the disease condition in the recipient mice after Cm infection with reduced Cm inclusion-forming units in lung tissues and significantly increased proportion of Th1 cells in lung and spleen tissues. Conclusions:Cm respiratory infection could induce the maturation and activation of DC, which promoted Th1 immune response. DC played an important role in Cm infection.

4.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-1029467

RESUMEN

Objective:To investigate the role of IL-21/IL-21R-mediated CD4 + T cells in Chlamydia muridarum ( Cm) respiratory infection. Methods:C57BL/6 mice (WT mice) and IL-21R -/- mice were used to establish the models of Cm respiratory infection through intranasal inhalation of Cm. Flow cytometry was used to detect the proportion, number, activity and function of CD4 + T cells in lung and spleen tissues at 0, 3, 7 and 14 d after Cm respiratory tract infection. IFN-γ and IL-4 levels in spleen cell culture supernatants were detected by ELISA. Na?ve WT mice were transferred with CD4 + T cells in the spleen tissues of IL-21R -/- mice or WT mice on 7 d after infection and given Cm intranasally 2 h later. Then the mice were weighed daily and sacrificed on 14 d after infection. The bacterial load and pathological changes in lung were analyzed. Flow cytometry was performed to detect the proportions and numbers of neutrophils (CD45 + CD11b + Gr-1 high) and alveolar macrophages (CD45 + F4/80 + CD11c high)as well as the proportions of Th1 (IFN-γ + CD4 + ) and Th2 (IL-4 + CD4 + ) cells. ELISA was also performed to measure IFN-γ and IL-4 levels in spleen cell culture supernatants. Results:Compared with WT mice, IL-21R -/- mice showed elevated numbers and enhanced activation of CD4 + T cells, increased proportion of Th1 cells and decreased proportion of Th2 cells in spleen and lung tissues after Cm respiratory infection. Besides, IFN-γ levels increased, while IL-4 levels decreased in spleen cell culture supernatants of IL-21R -/- mice. After Cm infection, the na?ve WT transferred with CD4 + T cells from IL-21R -/- mice showed less body weight loss, reduced bacterial load and alleviated pathological changes in lung tissues, increased proportion of Th1 cells in lung tissue and higher IFN-γ level in spleen cell culture supernatants. Conclusions:IL-21/IL-21R-mediated CD4 + T cells could aggravate Cm respiratory infection by suppressing Th1 cell immune responses.

5.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-934032

RESUMEN

Objective:To investigate the infiltration and polarization of macrophages in mice during Chlamydia muridarum ( Cm) respiratory infection. Methods:C57BL/6 mice were intranasally infected with 1×10 3 inclusion-forming units (IFU) of Cm to establish the mouse model of Cm respiratory tract infection. The percentages of CD45 + F4/80 + cells and the macrophages expressing CD86, major histocompatibility complex Ⅱ (MHC), inducible nitric oxide synthase (iNOS) and CD206 were detected by flow cytometry. Expression of iNOS, CD206 and CCL2 at mRNA level was detected by real-time quantitative PCR. Results:Cm respiratory tract infection induced the increase of macrophages in mouse lung tissues. Compared with uninfected group, CD45 + F4/80 + macrophages were increased significantly from day 3 and reached the peak on day 7 after Cm infection. Moreover, the expression of CD86, MHCⅡ and CCL2 was increased, and the macrophages were polarized to M1 phenotype. However, the expression of M2 macrophage marker CD206 was decreased gradually. Further studies showed that iNOS expression, the indicator of M1 macrophage activation, was increased after Cm infection and reached to the top on day 7. Conclusions:Cm respiratory infection could induce the infiltration of macrophages in lung tissues and promote the polarization of macrophages to M1 phenotype.

6.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-958230

RESUMEN

Objective:To investigate the effects of Chlamydia muridarum ( Cm) respiratory tract infection on the infiltration and polarization of alveolar macrophages (AMs) and pulmonary interstitial macrophages (IMs). Methods:A C57BL/6 mouse model of Cm respiratory tract infection was established through nasal inhalation. Flow cytometry was used to detect AMs (CD45 + F4/80 + CD11c + ) and IMs (CD45 + F4/80 + CD11c -) in lung tissues at 0, 3, 7 and 14 d after Cm respiratory tract infection. The proportions of M1 (CD80 + , CD86 + , MHCⅡ + , iNOS + ) and M2 (CD206 + , Arg1 + ) macrophages in AMs and IMs were also detected. Results:(1) Cm respiratory tract infection induced the infiltration of AMs and IMs. Compared with the uninfected group (0 d), the proportions and the numbers of AMs and IMs of were significantly increased 3 d after infection ( P<0.05, P<0.01). The numbers of AMs and IMs reached the peak 7 d after infection ( P<0.001). (2) Compared with the uninfected group, the proportions of CD80 + and CD86 + cells in AMs were significantly up-regulated 3 d after infection ( P<0.05, P<0.01); the proportion of MHCⅡ + cells in AMs increased after infection and reached the peak at 14 d ( P<0.05), while the proportion of CD206 + cells decreased after infection ( P<0.05). (3) Compared with the uninfected group, the proportions of CD80 + and CD86 + cells in IMs were increased 3 d after infection ( P<0.05, P<0.001) and the proportion of MHCⅡ + cells was significantly increased 14 d after infection ( P<0.01), while there was no significant change in the proportion of CD206 + cells. (4) In AMs, the proportion of iNOS + cells increased continuously after infection ( P<0.01), while the proportion of Arg1 + cells decreased continuously after infection, especially at 7 d and 14 d ( P<0.05). In IMs, the proportion of iNOS + cells reached the peak at 7 d ( P<0.001), but the proportion of Arg1 + cells showed no significant change after infection. Conclusions:Cm respiratory tract infection induced the infiltration of AMs and IMs, stimulated the polarization of AMs and IMs towards the M1 phenotype and weakened the polarization of AMs to M2 macrophages, but had no significant influence on the polarization of IMs towards the M2 phenotype.

7.
Artículo en Inglés | WPRIM (Pacífico Occidental) | ID: wpr-889300

RESUMEN

Purpose@#This study was aimed to evaluate the clinical significance and prognostic value of CRP/albumin ratio (CAR) in patients with gastric cancer. @*Methods@#The data of 205 gastric cancer patients who underwent surgery was analyzed retrospectively. The association of CAR with the clinical features and prognostic value in gastric cancer was analyzed. The data of this study was combined with previous studies to further determine the prognostic value of CAR in patients with gastric cancer using a metaanalysis method. @*Results@#Cox analysis revealed that preoperative CAR was an independent prognosis indicator in patients with gastric cancer. High expression of CAR indicated a shorter survival time than in those with lower expression. CAR has a higher prognostic value in the 1-, 3-, and 5-year overall survival in patients with gastric cancer. CAR showed significant difference regarding the gastric cancer patients’ age, M stage, and clinical stage. The discriminate value of CAR in M stage of gastric cancer was high (area under the curve, 0.809). A meta-analysis combining previous data and our data showed that preoperative CAR demonstrated a significant association with the overall survival of patients with gastric cancer. @*Conclusion@#This study demonstrated that preoperative CAR could serve as an important prognostic indicator in patients with gastric cancer.

8.
Artículo en Inglés | WPRIM (Pacífico Occidental) | ID: wpr-897004

RESUMEN

Purpose@#This study was aimed to evaluate the clinical significance and prognostic value of CRP/albumin ratio (CAR) in patients with gastric cancer. @*Methods@#The data of 205 gastric cancer patients who underwent surgery was analyzed retrospectively. The association of CAR with the clinical features and prognostic value in gastric cancer was analyzed. The data of this study was combined with previous studies to further determine the prognostic value of CAR in patients with gastric cancer using a metaanalysis method. @*Results@#Cox analysis revealed that preoperative CAR was an independent prognosis indicator in patients with gastric cancer. High expression of CAR indicated a shorter survival time than in those with lower expression. CAR has a higher prognostic value in the 1-, 3-, and 5-year overall survival in patients with gastric cancer. CAR showed significant difference regarding the gastric cancer patients’ age, M stage, and clinical stage. The discriminate value of CAR in M stage of gastric cancer was high (area under the curve, 0.809). A meta-analysis combining previous data and our data showed that preoperative CAR demonstrated a significant association with the overall survival of patients with gastric cancer. @*Conclusion@#This study demonstrated that preoperative CAR could serve as an important prognostic indicator in patients with gastric cancer.

9.
J Sep Sci ; 44(2): 497-512, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33164302

RESUMEN

It is critical to investigate the adaptive development and the physiological mechanism of fish in external stimulation. In this study, the response of Barbus capito to salinity-alkalinity exposure was explored by high-throughput nontargeted and liquid chromatography-mass spectrometry-based metabolomics to investigate metabolic biomarker and pathway changes. Meanwhile, the biochemical indexes of Barbus capito were measured to discover the chronic impairment response to salinity-alkalinity exposures. A total of 29 tissue metabolites were determined to deciphering the endogenous metabolic changes of fishes during the different concentration salinity-alkalinity exposures environment, which were mainly involved in the key metabolism including the phenylalanine, tyrosine, and tryptophan biosynthesis, arachidonic acid metabolism, pyruvate metabolism, citrate cycle, and glycerophospholipid metabolism. Finally, we found the amino acid metabolism as key target was associated with the endogenous metabolites and metabolic pathways of Barbus capito to salinity-alkalinity exposures. In conclusion, metabolomics is a potentially powerful tool to reveal the mechanism information of fish in various exposure environments.


Asunto(s)
Ensayos Analíticos de Alto Rendimiento , Metabolómica , Bicarbonato de Sodio/química , Cloruro de Sodio/química , Animales , Biomarcadores/análisis , Biomarcadores/metabolismo , Cromatografía Liquida , Cyprinidae , Espectrometría de Masas , Salinidad
10.
Trials ; 21(1): 645, 2020 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-32665034

RESUMEN

BACKGROUND: Delirium is an acute status of brain dysfunction that commonly occurs in patients who have undergone cardiac surgery, and increases morbidity and mortality. It is associated with risk factors, such as older age, use of narcotics, cardiopulmonary bypass, and hypothermia. Dexmedetomidine infusion might exert a neuroprotective effect. However, the effect of perioperative administration of dexmedetomidine on the incidence of postoperative delirium (POD) in patients undergoing cardiac or non-cardiac surgery is yet controversial. The present study aimed to reveal the effect of intraoperative dexmedetomidine administration on the incidence of delirium in adult patients following cardiac surgery. METHODS: This single-center, randomized, double-blinded, and placebo-controlled trial consisted of 652 patients randomly divided into two groups: dexmedetomidine and placebo. 0.6 µg/kg dexmedetomidine will be infused 10 min after central vein catheterization, followed by a continuous infusion at a speed of 0.4 µg/kg/h until the end of surgery in the dexmedetomidine group, while normal saline will be administered at the same rate in the placebo group. The primary outcome is the incidence of POD during the first 7 days post-surgery. The secondary outcomes include duration of mechanical ventilation after surgery, duration of stay in the intensive care unit and the hospital after surgery, incidence of hypotension during or after dexmedetomidine infusion, acute kidney injury and sudden arrhythmia during the hospital stay postoperatively, and all-cause mortality in 30 and 90 days after surgery, respectively. DISCUSSION: This study was approved by the Ethics Committee of the Chinese Academy of Medical Sciences Fuwai Hospital on 6 March 2019 (2019-1180). The results will be disseminated at academic conferences and submitted to peer-reviewed publications. Either positive or negative results will provide guidance for clinical practice. TRIAL REGISTRATION: The Chinese Clinical Trial Registry ( http://www.chictr.org.cn ) ChiCTR1900022583. Registered on 17 April 2019.


Asunto(s)
Procedimientos Quirúrgicos Cardíacos , Delirio/prevención & control , Dexmedetomidina/uso terapéutico , Fármacos Neuroprotectores/uso terapéutico , Adulto , Procedimientos Quirúrgicos Cardíacos/efectos adversos , Delirio/diagnóstico , Dexmedetomidina/administración & dosificación , Método Doble Ciego , Válvulas Cardíacas , Humanos , Estudios Prospectivos , Ensayos Clínicos Controlados Aleatorios como Asunto , Reproducibilidad de los Resultados , Volumen Sistólico , Función Ventricular Izquierda
11.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-905336

RESUMEN

Objective:To investigate the difference of isokinetic angle-specific moment curves between anterior cruciate ligament (ACL)-injured patients with and without patellofemoral cartilage injury (PFCI). Methods:A retrospective analysis was performed on patients underwent knee arthroscopy and isokinetic muscle strength testing before surgery from September, 2018 to September, 2019. Seventeen ACL-injured patients with PFCI and 17 ACL-injured patients without PFCI who matched in age, sex and meniscus injury were selected. Before arthroscopy, isometric and isokinetic strength of knee flexion and extension at velocity of 180°/s and 60°/s was tested by isokinetic dynamometer. Normalized torque-angle curves (torque/body mass) were generated in steps of 1° and the differences in angle-specific moment curves between two groups were compared. Results:At 180°/s, there was no significant difference in flexion isokinetic torque both healthy side and affected side between two groups (P >0.05); and no difference in extension torque of the healthy side (P >0.05), however, there was significant difference in extension torque of the affected side at 88° to 90° between two groups (t > 2.102, P <0.05). At 60°/s, there was significant difference in flexion torque of the healthy side at 62° to 82° between two groups (|t| >2.056, P <0.05), and no significant difference was found in flexion torque of the affected side (P >0.05), nor in extension torque of both sides between two groups (P > 0.05). A curve change was found at the beginning of the flexion and extension isokinetic moment curves at the velocity of 180°/s. The isometric knee extension torque was significantly different in the affected side between two groups (t = 2.858, P < 0.01), and no difference was found in isometric knee flexion torque in the affected side as well as both extension and flexion torques in the healthy side between two groups (t < 1.905, P > 0.05). Conclusion:The lower the isokinetic speed, the more significant the difference of strength is between ACL-injury patients with and without PFCI. High speed exercise is recommended for ACL-injured patients with PFCI.

12.
Chinese Journal of School Health ; (12): 1061-1065, 2020.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-823179

RESUMEN

Objective@#The status of masked obesity in female college students and the related factors were investigated to provide a theoretical reference for instructing the physical health education of female college students and improving their physical exercises.@*Methods@#Body composition tests were performed on 2 975 female college students, selected from 3 colleges in North China by using cluster sampling method during October to December in 2018, and their basic information and lifestyle were investigated using questionnaires.@*Results@#The incidence of masked obesity among female college students was 33.18%, of which 2.53% came from low-weight people. The difference in the incidence of masked obesity among female college students with different take-out frequencies was statistically significant (χ2=21.98, P<0.01). Compared with those who never take takeaway, people who take takeaway every day have an increased risk of masked obesity (OR=1.49, 95%CI=0.76-2.91). The difference in the incidence of masked obesity with the frequency of eating midnight snack was statistically significant (χ2=20.80, P<0.05). The difference in the incidence of masked obesity among female college students with different exercise time was statistically significant (χ2=18.49, P<0.01). Compared with exercise time above 60 min/d, female college students who are not exercising have an increased risk of masked obesity (OR=3.20, 95%CI=1.63-6.30). The difference in the incidence of masked obesity among female college students with different weight satisfaction was statistically significant (χ2=217.54, P<0.01). Compared with female college students who were satisfied with weight, female college students who were not satisfied with weight had an increased risk of masked obesity (OR=3.47, 95%CI=1.91-6.31). The difference in the incidence of masked obesity in different weightdown plans is statistically significant (χ2=186.40, P<0.01). Those who want to lose weight have a higher risk of developing masked obesity than those who want to gain weight (OR=18.11, 95%CI=5.54-50.13).@*Conclusion@#Female college students who drink a small amount of water, eat takeaways often, eat midnight snacks, do not exercise, and are not satisfied with their weight are more likely to develop masked obesity.

13.
RSC Adv ; 10(3): 1552-1571, 2020 Jan 07.
Artículo en Inglés | MEDLINE | ID: mdl-35494719

RESUMEN

The aims of this study is to explore the metabolomic biomarker and pathway changes in crucian under carbonate alkalinity exposures using high-throughput metabolomics analysis based on ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-tandem mass spectrometry (UPLC-ESI-QTOF-MS) for carrying out adaptive evolution of fish in environmental exposures and understanding molecular physiological mechanisms of saline-alkali tolerance in fishes. Under 60 day exposure management, the UPLC-ESI-QTOF-MS technology, coupled with a pattern recognition approach and metabolic pathway analysis, was utilized to give insight into the metabolic biomarker and pathway changes. In addition, biochemical parameters in response to carbonate alkalinity in fish were detected for chronic impairment evaluation. A total of twenty-seven endogenous metabolites were identified to distinguish the biochemical changes in fish in clean water under exposure to different concentrations of carbonate alkalinity (CA); these mainly involved amino acid synthesis and metabolism, arachidonic acid metabolism, glyoxylate and dicarboxylate metabolism, pyruvate metabolism and the citrate cycle (TCA cycle). Compared with the control group, CA exposure increased the level of blood ammonia; TP; ALB; Gln in the liver and gills; GS; urea in blood, the liver and gills; CREA; CPS; Glu and LDH; and decreased the level of weight gain rate, oxygen consumption, discharge rate of ammonia, SOD, CAT, ALT, AST and Na+/K+-ATPase. At low concentrations, CA can change the normal metabolism of fish in terms of changing the osmotic pressure regulation capacity, antioxidant capacity, ammonia metabolism and liver and kidney function to adapt to the CA exposure environment. As the concentration of CA increases, various metabolic processes in crucian are inhibited, causing chronic damage to the body. The results show that the metabolomic strategy is a potentially powerful tool for identifying the mechanisms in response to different environmental exposomes and offers precious information about the chronic response of fish to CA.

14.
Mol Immunol ; 116: 180-190, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31704501

RESUMEN

Infectious pancreatic necrosis virus (IPNV) and infectious hematopoietic necrosis virus (IHNV) are two common viral pathogens that cause severe economic losses in all salmonid species in culture, but especially in rainbow trout. Although vaccines against both diseases have been commercialized in some countries, no such vaccines are available for them in China. In this study, a recombinant virus was constructed using the IHNV U genogroup Blk94 virus as a backbone vector to express the antigenic gene, VP2, from IPNV via the reverse genetics system. The resulting recombinant virus (rBlk94-VP2) showed stable biological characteristics as confirmed by virus growth kinetic analyses, pathogenicity analyses, indirect immunofluorescence assays and western blotting. Rainbow trout were immunized with rBlk94-VP2 and then challenged with the IPNV ChRtm213 strain and the IHNV Sn1203 strain on day 45 post-vaccination. A significantly higher survival rate against IHNV was obtained in the rBlk94-VP2 group on day 45 post-vaccination (86%) compared with the PBS mock immunized group (2%). Additionally, IPNV loads decreased significantly in the rBlk94-VP2 immunized group in the liver (28.6-fold to 36.5-fold), anterior kidney (21.7-fold to 44.2-fold), and spleen (14.9-fold to 22.7-fold), as compared with the PBS mock control group. The mRNA transcripts for several innate and adaptive immune-related proteins (IFN-γ, IFN-1, Mx-1, CD4, CD8, IgM, and IgT) were also significantly upregulated after rBlk94-VP2 vaccination, and neutralizing antibodies against both IHNV and IPNV were induced on day 45 post-vaccination. Collectively, our results suggest that this recombinant virus could be developed as a vaccine vector to protect rainbow trout against two or more diseases, and our approach lays the foundations for developing live vaccines for rainbow trout.


Asunto(s)
Enfermedades de los Peces/inmunología , Virus de la Necrosis Hematopoyética Infecciosa/inmunología , Oncorhynchus mykiss/inmunología , Oncorhynchus mykiss/virología , Animales , Anticuerpos Antivirales/inmunología , Infecciones por Birnaviridae/inmunología , Infecciones por Birnaviridae/virología , China , Riñón Cefálico/inmunología , Riñón Cefálico/virología , Virus de la Necrosis Pancreática Infecciosa/inmunología , Pancreatitis Aguda Necrotizante/inmunología , Pancreatitis Aguda Necrotizante/virología , Infecciones por Rhabdoviridae/inmunología , Infecciones por Rhabdoviridae/virología , Bazo/inmunología , Bazo/virología , Vacunación/métodos , Vacunas de ADN/inmunología , Carga Viral/métodos , Vacunas Virales/inmunología
15.
Arch Virol ; 164(10): 2505-2513, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31377888

RESUMEN

Infectious hematopoietic necrosis virus (IHNV) was developed as a vector to aid the construction of vaccines against viral diseases such as viral hemorrhagic septicemia virus, spring viremia of carp virus, and influenza virus H1N1. However, the optimal site for foreign gene expression in the IHNV vector has not been determined. In the present study, five recombinant viruses with the green fluorescence protein (GFP) gene inserted into different genomic junction regions of the IHNV genomic sequence were generated using reverse genetics technology. Viral growth was severely delayed when the GFP gene was inserted into the intergenic region between the N and P genes. Real-time fluorescence quantitative PCR assays showed that the closer the GFP gene was inserted towards the 3' end, the higher the GFP mRNA levels. Measurement of the GFP fluorescence intensity, which is the most direct method to determine the GFP protein expression level, showed that the highest GFP protein level was obtained when the gene was inserted into the intergenic region between the P and M genes. The results of this study suggest that the P and M gene junction region is the optimal site within the IHNV vector to express foreign genes, providing valuable information for the future development of live vector vaccines.


Asunto(s)
Expresión Génica , Vectores Genéticos , Virus de la Necrosis Hematopoyética Infecciosa/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Fluorometría , Genes Reporteros , Proteínas Fluorescentes Verdes/biosíntesis , Proteínas Fluorescentes Verdes/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Genética Inversa
16.
J Virol Methods ; 265: 84-90, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30615899

RESUMEN

Reverse genetics systems are powerful tools for understanding the virulence mechanisms and gene functions of negative-sense RNA viruses. The reverse genetics systems commonly used for recombinant infectious hematopoietic necrosis virus (IHNV) are based on vaccinia virus infection. To avoid the potential biological safety risks associated with vaccinia virus, a recombinant IHNV virus strain Sn1203 (rIHNV-Sn1203) was rescued in this study using a mammalian cell line, BHK-21. The genome sequence authenticity of rIHNV-Sn1203 was confirmed using two silent genetic tags introduced by site-directed mutagenesis. Indirect immunofluorescence assays and transmission electron microscopy revealed that rIHNV-Sn1203 and wild-type IHNV-Sn1203 (wtIHNV-Sn1203) had identical immunogenicity and virion morphology. The virulence and pathogenicity of rIHNV-Sn1203 were assessed in vitro and in vivo. Although rIHNV-Sn1203 displayed trends toward delayed intracellular viral replication and lower virion yields compared with wtIHNV-Sn1203, statistical analyses revealed no significant differences between these two viruses. Moreover, rainbow trout challenged with rIHNV-Sn1203 and wtIHNV-Sn1203 showed indistinguishable mortality. Together, these results show that IHNV was successfully rescued using BHK-21 cells. This method is very convenient and may also be suitable for use in the recovery of other Novirhabdoviruses.


Asunto(s)
Virus de la Necrosis Hematopoyética Infecciosa/crecimiento & desarrollo , Genética Inversa/métodos , Virología/métodos , Animales , Línea Celular , Cricetinae , Enfermedades de los Peces/patología , Enfermedades de los Peces/virología , Técnica del Anticuerpo Fluorescente Indirecta , Virus de la Necrosis Hematopoyética Infecciosa/genética , Virus de la Necrosis Hematopoyética Infecciosa/patogenicidad , Virus de la Necrosis Hematopoyética Infecciosa/ultraestructura , Microscopía Electrónica de Transmisión , Oncorhynchus mykiss , Infecciones por Rhabdoviridae/patología , Infecciones por Rhabdoviridae/veterinaria , Infecciones por Rhabdoviridae/virología , Análisis de Supervivencia , Virus Vaccinia/genética , Virión/ultraestructura , Replicación Viral
17.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-857556

RESUMEN

OBJECTIVE To analyze the composition and function of N-glycoproteins in human plasma exosomes. METHODS Exosomes from human plasma were extracted by ultracentrifugation. The proteins from plasma exosomes were released by ultrasound and the obtained proteins were enzymatically degraded by trypsin to peptides. The N-glycopeptides in the mixture of the digested peptides was enriched by the hydrazide resin. The enriched N-glycoproteins were identified by mass spectrometry and subjected to gene ontology (GO) annotation analysis. RESULTS The plasma exosomes isolated by ultracentrifugation had a typical cup-shaped structure and the particle sizes were approximately 100 nm. Totally, 252 N-glycosylation sites corresponding to 131 N-glycoproteins were enriched and identified from exosomes of 1 mL human plasma digestion, which participated in many important biological processes, such as serine-type endopeptidase activity, serine-type endopeptidedase inhibitor acitivity and calcium ion binding. Sixty-seven of the N-glycoproteins had close relations with the occurrence of diseases. CONCLUSION Totally, 131 N-glycoproteins were identified in the research from the plasma exosome protein. These N-glycoproteins precipitate in many important biological processes and have close correlations with the occurrence and progression of various diseases.

18.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-790136

RESUMEN

Objective :To explore the feasibility and clinical value of real time three‐dimensional transesophageal echo‐cardiography (RT‐3D TEE) in diagnosis of congenital heart valvular disease (CHVD).Methods :A total of 135 CH‐VD patients treated in our hospital were selected .All patients received surgery ,and transthoracic echocardiography (TTE) and RT‐3D TEE inspection successively within 7d before surgery .Heart valve lesion condition was observed , and diagnostic results of two methods and surgical outcome were compared and analyzed .Results :RT‐3D TEE could display the morphological structure ,lesion degree and peripheral blood flow of heart valves in CHVD patients in a multi‐angle ,stereoscopic and clear way .It could find heart valve disease which is difficult to be identified by TTE , and corrected the diagnostic deviation .With surgical results as the gold standard ,diagnostic coincidence rate of RT‐3D TEE was significantly higher than that of TTE (97. 04% vs.91. 11%, P=0.039).CHVD diagnosed by RT‐3D TEE and TTE possessed a intermediate consistency (Kappa=0.477 , P=0. 001).Conclusion :RT‐3D TEE can pro‐vide more imaging information for the diagnosis of CHVD ,which can be used as an effective supplement for preop‐erative TTE examination .

19.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-1008244

RESUMEN

This research was performed to establish the HPLC fingerprint of Sabia parviflora. HPLC method was carried out on a Thermo Accucore-C18(4. 6 mm×150 mm,2. 6 μm) column by 30% tetrahydrofuran in methyl alcohol-acetonitrile-0. 1% phosphate solution as mobile phase at a flow rate of 1. 0 m L·min-1,the column temperature was 30 ℃ and the detection wavelength was 360 nm. The fingerprints were further evaluated by chemometrics methods including similarity analysis,hierarchical clustering analysis,and principal component analysis. In HPLC fingerprint,15 common peaks were selected as the common peaks,and 6 contents of them were identified. The similarity degrees of 38 batches of the samples was more than 0. 710,and the samples were divided into 6 clusters by their quality difference. The method was precision,repeatable,stable,simple and reliable,which could be used for quality control and evaluation of S. parviflora.


Asunto(s)
Cromatografía Líquida de Alta Presión , Análisis por Conglomerados , Medicamentos Herbarios Chinos , Análisis de Componente Principal , Control de Calidad
20.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-698494

RESUMEN

BACKGROUND: Non-surgical spinal decompression system can be used for the treatment of diseased intervertebral discs and avoid contraction and resistance of paravertebral muscle during traction. At present, there is lack of analysis of the efficacy of non-surgical spinal decompression in the treatment of lumbar disc herniation through surface electromyography. OBJECTIVE: To compare the effect of non-surgical spinal decompression system DRX9000 with conventional traction device in the treatment of lumbar disc herniation through surface electromyography. METHODS: Totally 60 patients with lumbar disc herniation were randomly divided into DRX9000 group and general traction group, with 30 cases in each group. The patients in DRX9000 group were treated with non-surgical spinal decompression system DRX9000 and the patients in general traction group were treated with conventional tractor. All patients underwent surface electromyography examination of erector spinal muscle and multifidus muscle before and after all treatment to evaluate the strength and fatigue of paravertebral muscle. The Visual Analogue Scale and Japanese Orthopaedic Association scores were used to assess the degree of pain and impairment due to lumbar disc herniation. RESULTS AND CONCLUSION: (1) After all of the treatment, the average electromyogram and mean power frequency slope of affected erector spinal muscle and multifidus muscle in both groups were increased than those before treatment (P < 0.05). The data of DRX9000 group were significantly higher than that of the general traction group (P < 0.05). (2) After all the treatment, the Visual Analogue Scale score decreased and the Japanese Orthopaedic Association score increased in both groups at the end of treatment compared with before treatment (P < 0.05). The Visual Analogue Scale score of DRX9000 group was significantly lower than that of the general traction group (P < 0.05) and the Japanese Orthopaedic Association score of DRX9000 group was significantly higher than that of the general traction group (P < 0.05). (3) These results showed that non-surgical spinal decompression system could effectively improve paravertebral muscle activity and muscle strength, lessen muscle fatigue, relieve pain, and improve function in patients with lumbar disc herniation, and its effect is obviously better than that of general traction.

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