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The harm of plant virus disease is serious, which significantly restricts the sustainable development of agriculture and can cause huge economic losses. Monitoring plant health and early detection of viral pathogens are essential to reduce the spread of disease. Therefore, in order to realize the early detection of plant viral diseases in the field, a sensitive, specific and efficient colorimetric visual technique for plant viral RNA was designed by combining the colorimetric method based on gold nanoparticles (AuNPs) and hybrid chain reaction (HCR). In this study, tobacco mosaic virus (TMV) was used as a model to design two hairpin structures H1/ H2 with single-stranded tails based on TMV-specific conserved fragments. TMV could open the hairpin structure to alternately form long double-straight DNA. The binding difference between AuNPs and two nucleic acid states before and after HCR reaction resulted in colorimetric signal generation, thus realizing visual detection of TMV. After optimizing the concentration of Tris-HAc, the concentration of hairpin structure and the reaction time of HCR, the best detection conditions were obtained. The sensitivity and specificity of the technique were analyzed and real samples were tested under optimal conditions. The results showed that the absorbance ratio of AuNPs (A
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We performed an extensively targeting metabolomic detecting using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) to compare the secondary metabolites in Dang shen [Codonopsis pilosula (Franch.) Nannf.] from Shanxi and Gansu provinces. The findings showed that 161 secondary metabolites in 6 groups (phenolic acids, flavonoids, lignans and coumarins, alkaloids, terpenoids, others) were found from Dang shen in Changzhi city of Shanxi province and Dingxi city of Gansu province. There were 98 secondary metabolites which is differed significantly. In comparison to Dingxi city, 33 different secondary metabolites of Dang shen in Changzhi city had a greater relative content, whereas relative content of 65 different metabolites in Dingxi city was higher. Metabolic pathway enrichment analysis revealed that phenolic acids and flavonoids were significantly different in the secondary metabolites of Dang shen from different producing places. This may be one of the reasons for the difference in the quality of Dang shen in Shanxi and Gansu provinces. This work compared and analyzed the secondary metabolites of Dang shen from Dingxi city in Gansu province and Changzhi city in Shanxi province for the first time, which lays the foundation for further study on the quality of Dang shen.
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Soybean mosaic virus (SMV) is a critical pathogen that reduces the soybean yield and seed quality worldwide, and SMV has a restricted natural host range. In this study, we used sequence-independent amplification (SIA) methods to identify the viruses that may cause the Atractylodes macrocephala Koidz disease. Results revealed that there is SMV in diseased Atractylodes macrocephala leaves, and we named this isolate as SMV-Am. To further characterize the genomic structural and phylogenetic relationships of SMV-Am, genomic dsRNA was extracted, and the genomic sequence was amplified by RT-PCR and RACE. The results of bioinformatics analysis showed that the genomic RNA of SMV-Am is 9587 nucleotides in length, which conforms to the typical characteristics of Potyvirus. According to the sequence alignment of complete nucleotide sequences, SMV-Am showed the highest level of nucleotide homology and amino acid sequences to SMV-Liaoning, 96. 57% and 98. 86%, respectively. In addition, phylogenetic analysis based on the nucleotide sequences of other SMV isolates of SMV-Am revealed that SMV-Am was most closely related to SMV-Liaoning. Then, the SMV-Am protein was further analyzed by I-TASSER and PyMOL software, revealing that the key amino acid mutations led to the structural changes of P1, HC-Pro, P3, 6K2, NIa-pro and NIb proteins, with P1 the most obvious. Finally, recombination has been detected at the position of 6560-8950 nucleotides, the main parent is the isolate SMV-XFQ012 (accession number KP710875. 1), and the secondary parent is isolate SMV-pCB301-SC15 (accession number MH919386. 1). This study is the first report of SMV in Atractylodes macrocephala Koidz, and these results are expected to provide a scientific basis for the prevention and control of SMV infection on Atractylodes macrocephala Koidz.
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Plant virus diseases are one of the major diseases restricting erop production.Timely identification of their pathogen and development rules is the prerequisite for effective control of their large- scale spread.However, long cycle, tedious steps and strict detection environment were the disadvantages existing in the detection technology of plant virus disease.In this study, Tobacco Mosaic virus (TMV) was used as a model to he extract UNA based on CMBs-ACPtmv , which was design based on the principle of complementary base pairing.Meanwhile, the experimental conditions were optimized and analyzed, including the preparation conditions of functionalized magnetic beads, the reaction conditions during extraction, and the sensitivity, stability and other properties of the method.The results showed the ability to capture RNA of CMBs-ACPtmv were best when prepared with 4 fxmol capture probe (ACPTMV ) and 0.08 mg carboxyl magnetic beads (CMBs) ; After 3 min of extraction, CMBs-ACPtmv has the best RNA extraction effect, but when the extraction temperature of CMBs-ACPtmv was changed, its extraction capacity showed no significant change; In the comprehensive performance evaluation, the sensitivity of CMBs-ACPjjjv can reach 2.5 ng/fxL, and the detection stability is good.Compared with conventional RNA extraction technology, CMBs-ACPimv has outstanding advantages in detection time and sample consumption.The functional magnetic beads extraction method established in this study is fast, safe and simple.It can achieve rapid extraction of plant virus RNA with simple equipment, which has a broad application prospect.
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Zucchini yellow mosaic virus (ZYMV) is a destructive cucurbit virus that causes extensive losses in the yield and quality of cucurbits (e.g., zucchini). At present, the analysis of genetic evolution of ZYMV is mainly based on CP sequence, and there has been little progress in addressing its phylogenetic relationships and evolution at the whole genome level. In this study, the whole genome sequences of ZYMV isolated from Shanxi Wenshui (ZYMV-WS) and 109 other reported ZYMV isolates were used to explore their phylogenetic relationships. The results showed that ZYMV-WS had the highest level of nucleotide homology (94.35%) to ZYMV-SXSG, and the amino acid sequences of ZYMV-BR3 were identical (97.37%) to those of ZYMV-WS. In addition, the phylogenetic analysis of ZYMV revealed that all the isolates were clustered into two major branches, and its population structure assessment demonstrated a marked genetic differentiation between A4 and B. Furthermore, we also discovered the sites of specific amino-acid alteration, and their possible association with functional changes. These results will help to better understand the genetic diversity and phylogenetic process of ZYMV, and lay a theoretical foundation for the effective prevention and control of ZYMV infection in the near future.
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<p><b>Objective</b>To investigate the role of the serum inhibin B (INHB) level in evaluating the testicular function of the prepubertal patient with varicocele (VC) after high ligation of the spermatic vein (HLSV).</p><p><b>METHODS</b>This study included 31 prepubertal male patients with left VC, averaging 12.55 years of age and 9 complicated by right VC. We collected peripheral blood samples before and at 4, 12 and 26 weeks after HLSV as well as spermatic venous blood samples intraoperatively for determination of the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), anti-sperm antibody (AsAb) and serum INHB by ELISA.</p><p><b>RESULTS</b>Compared with the baseline, statistically significant differences were observed in the INHB level in the peripheral blood at 12 and 26 weeks after operation (［255.18 ± 69.97］ vs ［141.78 ± 59.82］ pg/ml, P < 0.05) and that in the spermatic venous blood intraoperatively (［255.18 ± 69.97］ vs ［412.44 ± 259.42］ pg/ml, P < 0.01). Spearman's analysis showed a negative correlation between the level of INHB and that of FSH (r = －0.224, P < 0.01).</p><p><b>CONCLUSIONS</b>The level of serum INHB in the peripheral blood of the prepubertal VC patient is decreased within 6 months after HLSV and negatively correlated with that of FSH. The levels of INHB and FSH may well reflect the testicular function of the prepubertal VC patient.</p>
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Adolescente , Niño , Humanos , Masculino , Anticuerpos , Sangre , Biomarcadores , Sangre , Hormona Folículo Estimulante , Sangre , Inhibinas , Sangre , Hormona Luteinizante , Sangre , Espermatozoides , Alergia e Inmunología , Testosterona , Sangre , Varicocele , SangreRESUMEN
BACKGROUND:Hidden blood loss is an important risk for the intertrochanteric fracture patients, especial y the elderly patients, which can cause anemia in patients after internal fixation and can affect wound healing and patient recovery. OBJECTIVE:To compare the perioperative hidden blood loss and the risk factors of proximal femoral anti-rotation intramedul ary nail internal fixation and dynamic hip screw fixation for the treatment of femoral intertrochanteric fracture. METHODS:We selected 70 cases of femoral intertrochanteric fracture patients who treated with proximal femoral anti-rotation intramedul ary nail and dynamic hip screw fixation, including 21 patients with the age ≥ 80 years and 49 patients with the age30 kg/m2 and 42 patients with the body mass index ≤ 30 kg/m2;30 patients received anti-rotation intramedul ary nail internal fixation and 40 patients received dynamic hip screw fixation. The perioperative blood loss was calculated with Gross formula according to the changes of height, body mass index and the hematocrit before and after fixation. RESULTS AND CONCLUSION:The mean total blood loss was 936 mL, the mean dominant blood loss was 237 mL and the mean hidden blood loss was 699 mL. The hidden blood loss was accounted for 74.7%in total blood loss. The dominant blood loss in the dynamic hip screw fixation group was higher than that in the anti-rotation intramedul ary nail internal fixation group, and the hidden blood loss was lower than the anti-rotation intramedul ary nail internal fixation group. The total blood loss and the hidden blood loss of the elderly patients were higher than those of the non-elderly patients;there was no significant difference between male and female patients, obesity and normal patients. The results indicate that hidden blood loss is the major reason for total blood loss of femoral intertrochanteric fracture after internal fixation. The hidden blood loss of anti-rotation intramedul ary nail internal fixation is larger than that of dynamic hip screw fixation, and elder is the risk factor for hidden blood loss.
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MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.
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<p><b>OBJECTIVE</b>To explore the relationship between Helicobacter pylori (Hp) infection and iron status using serum ferritin (SF) as a marker for total iron and to identify the related factors of iron nutritional status among preschool children.</p><p><b>METHODS</b>By cluster sampling, we recruited 475 preschool children aged 2 to 7 years. A structured questionnaire and diet form were sent to the parents of these children to obtain related information about the socioeconomic level and dietary intakes. After collecting blood samples, the following indexes were measured. Hp IgG antibodies were measured with a dot enzyme-linked immunoassay; hemoglobin, Hct, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width index (RDW) with automatic Complete Blood Count; SF with an immunoradiometric assay. Stool Hp antigen and occult bleeding were measured with ELISA among individuals who were Hp seropositive. Hp status was defined as positive when both serum and stool antigen tests were positive, Hp status was defined as negative when serum antigen test was negative; 24-hour weighting and recording methods were used to dietary survey for three days in May and December 2001, respectively, dietary intakes including energy, protein and micronutrient were calculated using nursery school nutrition software and evaluated by Chinese Dietary Reference Intakes (DRIs). Mann-Whitney test was used to compare mean ranks of SF in Hp-positive and Hp-negative children. To obtain an adjusted estimate of the impact of Hp infection on SF, a multivariate analysis of covariance was done to evaluate the different level of SF between Hp infected and non-infected status. The relationship between iron deficiency and gender, age, socioeconomic condition, iron intake, and calcium intake was assessed by univariate analysis. An unconditional multivariate logistic regression analysis was also performed. Iron deficiency status was dichotomized and placed as the dependent variable. Hp infection status was incorporated together with possible confounding factors as independent variables in a final logistic regression model. All the data were managed by EPI Info 5.01a and analyzed by SAS (Version 6.12).</p><p><b>RESULTS</b>Totally 64 children were diagnosed as Hp-positive and 305 as Hp-negative. Mann-Whitney test and multivariate analysis of covariance both showed that SF concentration was significantly lower in Hp infected individuals than non-infected individuals. Adjusted mean level and 95% confidence interval of SF in infected and non-infected children was 23.62 microg/L (7.13 microg/L-78.26 microg/L), 33.48 microg/L (10.28 microg/L-109.06 microg/L), respectively. The relationship between Hp infection and iron deficiency status persisted in logistic regression analysis after adjusting for possible confounding factors (OR: 7.95; 95% CI 2.56 - 24.67).</p><p><b>CONCLUSION</b>Iron nutritional status was reduced in Hp infected preschool children. Hp infection appears to be an independent risk factor or an added stressor on iron status among preschool children.</p>
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Niño , Preescolar , Femenino , Humanos , Masculino , Anticuerpos Antivirales , Sangre , Índices de Eritrocitos , Ferritinas , Sangre , Infecciones por Helicobacter , Sangre , Helicobacter pylori , Alergia e Inmunología , Hemoglobinas , Modelos Logísticos , Análisis Multivariante , Evaluación NutricionalRESUMEN
Objective To investigate the protective effects of Ginkgo Biloba leaves(EGb) preconditioning on liver graft in rat liver transplantation.Methods Male Sprague-Dawley rats were used as donors and(recipients) of orthotopic liver transplantation(OLT).The rats were randomly divided into EGb group,normal saline(NS) control group and sham operation(SO) group.The animals were killed at 2h,6h,24h after graft reperfusion.Plasma samples were collected for ALT and AST test.Liver tissues were collected to detect the expression of TNF-?mRNA and Bcl-2mRNA by RT-PCR.Also,liver tissues were used to detect rat(liver) histological change and apoptosis by TUNEL.Results The serum levels of ALT in EGb group were(significantly) lower than the NS group(P