Identification, expression and DNA variation analysis of high affinity nitrate transporter NRT2/3 gene family in Sorghum bicolor / 生物工程学报

Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.

Identification and expression analysis of NRT1 family genes in Rehmannia glutinosa / 中国中药杂志

NRT1 family proteins play an important roles for absorbing and transporting of nitrate in different plants. In order to identify the NRT1 family genes of Rehmannia glutinosa, this study used 11 NRT1 homologous proteins of Arabidopsis as probe sequences and aligned with the transcriptome data of R. glutinosa by using NCBI BLASTN software. Resulting there were 18 NRT1 proteins were identified in R. glutinosa. On basis of this, a series of the molecular characteristics of R. glutinosa NRT1 proteins including the conserved domains, the transmembrane structure, the subcellular location and phylogenetic features were in detail analyzed. At same time, it were systematically analyzed that the temporal and spatial expression patterns and characteristics of R. glutinosa NRT1 family genes in response to different stress factors. The results indicated that 18 R. glutinosa NRT1 family genes with the length of coding region from 1 260 bp to 1 806 bp, encoded proteins ranging from 419 to 601 amino acids, and all of they owned the domains of typical peptide transporter with 7 to 12 transmembrane domains. These R. glutinosa NRT1 family proteins mostly were found to locate on cellular plasma membrane, and belonged to the hydrophobic proteins. Furthermore, the evolutionary analysis found that the 18 R. glutinosa NRT1 protein family could be divided into two subfamilies, of which 14 NRT1 family genes might occur the positive selection, and 4 genes occur the passivation selection during the evolution process of R. glutinosa. In addition the expression analysis showed that 18 R. glutinosa NRT1 family genes have the distinct expression patterns in different tissues of R. glutinosa, and their expression levels were also obvious difference in response to various stress. These findings infield that 18 R. glutinosa NRT1 family proteins might have obviously different functional roles in nitrate transport of R. glutinosa. In conclusion, this study lays a solid theoretical foundation for clarifying the absorption and transport molecular mechanism of N element during R. glutinosa growth and development, and at same time for deeply studying the molecular function of R. glutinosa NRT1 proteins in absorption and transport of nitrate.

Emergence of dengue in tribal villages of Mandla district, Madhya Pradesh, India.

Background & objectives: Dengue (DEN) is a rapidly spreading arboviral disease transmitted by Aedes mosquitoes. Although it is endemic in India, dengue virus (DENV) infection has not been reported from tribal areas of Madhya Pradesh. Investigations were conducted to establish the aetiology of sudden upsurge of cases with febrile illness in June 2013 from tribal villages of Mandla district of Madhya Pradesh, India. Methods: The rapid response team of the National Institute for Research in Tribal Health, Jabalpur, conducted clinical investigations and field surveys to collect the samples from suspected cases. Samples were tested using molecular and serological tools. Collected mosquitoes were identified and tested for the presence of virus using semi nested reverse transcriptase-polymerase chain reaction (nRT-PCR). The sequences were analysed to identify serotype and genotype of the virus. Results: of the 648 samples collected from 18 villages of Mandla, 321 (49.53%) were found to be positive for dengue. The nRT-PCR and sequencing confirmed the aetiology as dengue virus type 2. Eighteen per cent of patients needed hospitalization and five deaths were attributed to dengue. The virus was also detected from Aedes aegypti mosquito, which was incriminated as a vector. Phylogenetic analysis revealed that the dengue virus 2 detected belonged to cosmopolitan genotype of the virus. Interpretation & conclusions: Dengue virus serotype 2 was detected as the aetiological agent in the outbreak in tribal villages of Mandla district of Madhya Pradesh. Conducive man-made environment favouring mosquitogenic conditions and seeding of virus could be the probable reasons for this outbreak. Urgent attention is needed to control this new threat to tribal population, which is already overburdened with other vector borne diseases.

Instructions of Neural Response Telemetry for Prediction of the Rehabilitation Effects and Mapping in Children Cochlear Implant / 听力学及言语疾病杂志

Objective To study the stability of NRT threshold and the correlation between NRT threshold and the value of behavior T ,C in order to instruct mapping in infant cochlear implant ,and to study whether the elic‐itation of NRT waveform can be applied to determine the effects of rehabilitation .Methods There were 159 cases with complete information in data in this study .150 out of the 159 cases coincided with the study requirements of the changes and correlation .We had collected the NRT thresholds of the electrodes of No .1 ,6 ,11 ,16 ,22 in switch-on ,one month after switched -on ,half a year and one year after switched -on .The analysis of variance method was adopted to compare the threshold changes of each electrode in different stages .We had collected the values of behavior T ,C in one year after switched -on .Pearson regression analysis was used to compare the correlation be‐tween NRT threshold in switch -on and this value of behavior T ,C .We applied MAIS assessment scale to make a questionnaire for all 159 patients during the follow -up at one year after switched -on .We had gathered the scores in three aspects ,the average score in questionnaire and the average hearing threshold of every child .All children were divided into four groups according to whether the elicitation of NRT waveform during - operation ,post-op‐eration .Results Regarding the NRT threshold ,there was no significant difference in the pairwise comparison of four time points(P>0 .05) .There was positive correlation between NRT threshold and behavior T ,C(P 0 .05) .Conclusion The NRT threshold remains steady from switched -on .A positive correlation exists between NRT threshold and the actual T ,C value .The NRT threshold can be used to estimate the T ,C value .But the accuracy of the measured value is less than the behavior method .We still need to use behavior method to measure as soon as possible .The NRT waveform appearance on the post-operation is more significant than that on the during -operation to prognosis of rehabilitation .

Comparison of apical transportation and change of working length in K3, NRT AND PROFILE rotary instruments using transparent resin block / 대한치과보존학회지

OBJECTIVES: The purpose of this study is to compare the apical transportation and working length change in curved root canals created in resin blocks, using 3 geometrically different types of Ni-Ti files, K3, NRT, and Profile. MATERIALS AND METHODS: The curvature of 30 resin blocks was measured by Schneider technique and each groups of Ni-Ti files were allocated with 10 resin blocks at random. The canals were shaped with Ni-Ti files by Crown-down technique. It was analyzed by Double radiograph superimposition method (Backman CA 1992), and for the accuracy and consistency, specially designed jig, digital X-ray, and CAD/CAM software for measurement of apical transportation were used. The amount of apical transportation was measured at 0, 1, 3, 5 mm from 'apical foramen - 0.5 mm' area, and the alteration of the working length before and after canal shaping was also measured. For statistics, Kruskal-Wallis One Way Analysis was used. RESULTS: There was no significant difference between the groups in the amount of working length change and apical transportation at 0, 1, and 3 mm area (p = 0.027), however, the amount of apical transportation at 5 mm area showed significant difference between K3 and Profile system (p = 0.924). CONCLUSIONS: As a result of this study, the 3 geometrically different Ni-Ti files showed no significant difference in apical transportation and working length change and maintained the original root canal shape.

Detection of nucleic acids in Borna disease virus in patients with viral encephalitis / 中华微生物学和免疫学杂志

Objective To explore the relationship between viral encephalitis(VE)and Borna disease virus(BDV)infection,and discuss the clinical features of viral encephalitis patients infected by BDV.Methods The p24 gene fragment of BDV in cerebrospinal fluid mononuclear cells(CSFMC)from 32 patients with viral encephaliris and 34 healthy individuals were examined by fluorescence quantitative nested reverse transcriptase polymerase chain reaction(FQ-nRT-PCR),in which, β-actin was used as internal reference. The clinical manifestations of patients with positive BDV p24 were watched. And the gene sequence,homology and amino acid sequence of positive products were analyzed. Results The positive rate(4/32,12.5%)of BDV p24 in CSFMC of patients with viral encephalitis was significantly higher than that of healthy individuals(0/34,0%),P＜0.05,and the numbers of copies of BDV p24 in CSFMC of VE patients were more than 102/μl. Compared with the sequences of the standard strain Ⅴ and the virus strain H1766 of BDV from horse supplied by GenBank,the homology of the gene sequence of positive product of CSFMC was 95.35% and 98.84% with gene mutation in 4 sites(nt1649 T→C, nt1656 G→A,nt1670 C→T, nt1676 C→T). The genetic relationship between the positive product and the virus strain BDV from horse was the nearest. The clinical features of VE patients with positive BDV p24 were mainly by mental and behavior disorders. Conclusion The occurrence of VE patients in the city of Zunyi in Guizhou province may be associated with BDV infection. The clinical features of VE patients with positive BDV p24 were mainly mental and behavior disorders.

The instrument-centering ability of four Nickel-Titanium instruments in simulated curved root canals / 대한치과보존학회지

The aim of this study was to evaluate the ability of newly marketed NRT instruments to maintain the original root canal configuration and curvature during preparation in comparison with the three existing instruments in simulated root canals. Simulated canals in resin blocks were prepared with ProFile, K3, ProTaper, and NRT instrument (n = 10 canals in each case). Pre- and post-operative images were recorded, and assessment of canal shape was completed with a computer image analysis program. The data were analyzed statistically using the One-way ANOVA followed by Duncan's test. The ability of instruments to remain centered in prepared canals at 1-, 2-mm levels was significantly better in ProFile groups than in other groups (p < 0.05). The change of centering ratio in NRT groups at 5-mm level was significantly greater than ProFile group and at 6- and 7-mm level than all other groups (p < 0.05). Although the NRT system was comparable to other systems in regards to its ability to maintain the canal configuration of apical portion, this system was more influenced by the mid-root curvature due to its stainless-steel files for coronal preflaring.

Intraoperative NRT Measurement During Cochlea Implant Surgery / 听力学及言语疾病杂志

Objective To explore the advantages of intraoperative NRT measurement(neural response telemetry)during cochlea implant surgery.Methods Intraoperative and postoperative NRT measurements were tested in ten SNHL children with CI24M-Nucleus and four SNHL children with CI24R(CS)-Nucleus, the ECAP threshold of intraoperative and postoperative measurements were compared, and the first maps were set up according to the intraoperative ECAP threshold.Results The average intraoperative ECAP threshold of 22 electrodes was higher than postoperative ECAP threshold, the average intraoperative ECAP threshold of CI24M-Nucleus' 22 electrodes was 11CL greater than postoperative ECAP threshold. It was just five minutes to record ECAP of 10 electrodes, and the beginning of fitting was finished in one hour.Conclusion It is confirmed that intraoperative NRT measurement could test the integrity of implants, and could estimate the first map. It is convenient to use and should be popularized.

Influence of Changing the Parameters on the Wave of NRT / 听力学及言语疾病杂志

Objective To inevestigate the changes of the responses by changing the parameters in the neural response tetemety(NRT).Methods In this project,twenty patients who were implanted in Nucleus C124M were measured the electrically evoked compound action potential(ECAP)using the NRT system.It was expected to observe the changes of the responses by changing the parameters in the NRT 2.04 software.Results It was found that the parameters that made great influences to the responses were current level,stimulate pulse width,gain,delay and masker advance.Conclusion The parameters that made great influences to the record time were stimulate rate and number of sweeps.

The Study of EABR and ECAP in Cochlear Implantation / 听力学及言语疾病杂志

Objective To explore the threshold and latency of patients receiving cochlear implantation using electrically evoked auditory brainstem responses (EABR), and to evaluate the significance of EABR applied to those patients.Methods The EABR and ECAP were recorded in 14 subjects who were operated for Nucleus24 cochlear implant.They were assigned to Group A and Group B according to the results of ECAP. The threshholds of EABR, the wave latency of III and V and inter-wave latency of III-V of EABR were compared and analyzed between Group A and Group B.Results The threshholds of EABR were higher in Group B than in Group A on electrode No 22 when the pulse width was 25,50,75,100,125 ?s,respectively. There were significant differences between Group A and Group B(P0.05).Conclusion The threshholds of EABR were lower in patients whose ECAP findings were positive. The latencies of III and V,and inter-wave latency of III–V of EABR had no significant difference according to the positive findings of ECAP.