Comparative Transcriptome Profiling Reveals Two WRKY Transcription Factors Positively Regulating Polysaccharide Biosynthesis in Polygonatum cyrtonema.

Polygonatum cyrtonema (P. cyrtonema) is a valuable rhizome-propagating traditional Chinese medical herb. Polysaccharides (PCPs) are the major bioactive constituents in P. cyrtonema. However, the molecular basis of PCP biosynthesis in P. cyrtonema remains unknown. In this study, we measured the PCP contents of 11 wild P. cyrtonema germplasms. The results showed that PCP content was the highest in Lishui Qingyuan (LSQY, 11.84%) and the lowest in Hangzhou Lin'an (HZLA, 7.18%). We next analyzed the transcriptome profiles of LSQY and HZLA. Through a qRT-PCR analysis of five differential expression genes from the PCP biosynthesis pathway, phosphomannomutase, UDP-glucose 4-epimerase (galE), and GDP-mannose 4,6-dehydratase were determined as the key enzymes. A protein of a key gene, galE1, was localized in the chloroplast. The PCP content in the transiently overexpressed galE1 tobacco leaves was higher than in the wild type. Moreover, luciferase and Y1H assays indicated that PcWRKY31 and PcWRKY34 could activate galE1 by binding to its promoter. Our research uncovers the novel regulatory mechanism of PCP biosynthesis in P. cyrtonema and is critical to molecular-assisted breeding.

[Identification and expression of uridine diphosphate glycosyltransferase(UGT) gene family from Dendrobium officinale].

Uridine diphosphate glycosyltransferase(UGT) is a highly conserved protein in plants, which usually functions in secondary metabolic pathways. This study used the Hidden Markov Model(HMM) to screen out members of UGT gene family in the whole genome of Dendrobium officinale, and 44 UGT genes were identified. Bioinformatics was used to analyze the structure, phylogeny, and promoter region components of D. officinale genes. The results showed that UGT gene family could be divided into four subfamilies, and UGT gene structure was relatively conserved in each subfamily, with nine conserved domains. The upstream promoter region of UGT gene contained a variety of cis-acting elements related to plant hormones and environmental factors, indicating that UGT gene expression may be induced by plant hormones and external environmental factors. UGT gene expression in different tissues of D. officinale was compared, and UGT gene expression was found in all parts of D. officinale. It was speculated that UGT gene played an important role in many tissues of D. officinale. Through transcriptome analysis of D. officinale mycorrhizal symbiosis environment, low temperature stress, and phosphorus deficiency stress, this study found that only one gene was up-regulated in all three conditions. The results of this study can help understand the functions of UGT gene family in Orchidaceae plants and provide a basis for further study on the molecular regulation mechanism of polysaccharide metabolism pathway in D. officinale.

To investigate the correlation between normal fetal biventricular myocardial function and gestational age using velocity vector imaging.

Objective: The aim of this study was to evaluate the left and right ventricular segmental and global myocardial function of normal fetuses using velocity vector imaging and explore the correlation between global myocardial function parameters and gestational age. Methods: A total of 127 normal fetuses were selected and divided into five groups according to gestational age for the measurement of their left and right ventricular segmental and global velocity, strain, and strain rate. This study also explored the change trend in the global myocardial function parameters at different gestational ages and analyzed its correlation with gestational age. Results: The peak velocities of the biventricular segments of the normal fetuses showed a decreasing trend from the basal to the middle to the apex segment, and the differences were statistically significant (P < 0.05). However, the strain and peak strain rate between adjacent segments showed no significant differences (P > 0.05). The peak global velocity of both ventricles increased with the gestational age, and it was moderately correlated with gestational age; however, the correlation of strain and peak strain rate with gestational age was not statistically significant (P > 0.05). Conclusion: In normal fetuses, the peak myocardial velocity of the biventricular segments showed a decreasing trend from the basal to the apical segment. The global peak myocardial velocity was linearly correlated with gestational age; however, the global strain and peak strain rate did not change as gestational age increased, indicating that the myocardial deformability of the fetus' ventricles was constant in the middle and late trimesters.

Integrating Transcriptomics and Hormones Dynamics Reveal Seed Germination and Emergence Process in Polygonatum cyrtonema Hua.

Polygonatum cyrtonema Hua is a traditional Chinese herb propagated using rhizomes, and excessive demand for seedlings and quality deterioration caused by rhizome propagation has highlighted that seed propagation may be an ideal solution to address these issues. However, the molecular mechanisms involved in P. cyrtonema Hua seed germination and emergence stages are not well understood. Therefore, in the present study, we performed transcriptomics combined with hormone dynamics during different seed germination stages, and 54,178 unigenes with an average length of 1390.38 bp (N50 = 1847 bp) were generated. Significant transcriptomic changes were related to plant hormone signal transduction and the starch and carbohydrate pathways. Genes related to ABA(abscisic acid), IAA(Indole acetic acid), and JA(Jasmonic acid) signaling, were downregulated, whereas genes related to ethylene, BR(brassinolide), CTK(Cytokinin), and SA(salicylic acid) biosynthesis and signaling were activated during the germination process. Interestingly, GA biosynthesis- and signaling-related genes were induced during the germination stage but decreased in the emergence stage. In addition, seed germination significantly upregulated the expression of genes associated with starch and sucrose metabolism. Notably, raffinose biosynthesis-related genes were induced, especially during the emergence stage. In total, 1171 transcription factor (TF) genes were found to be differentially expressed. Our results provide new insights into the mechanisms underlying P. cyrtonema Hua seed germination and emergence processes and further research for molecular breeding.

Physical education movement and comprehensive health quality intervention under the background of artificial intelligence.

The application of artificial intelligence has realized the transformation of people's production and lifestyle, and also promoted the progress of physical education and comprehensive health quality. The application of artificial intelligence in the current physical education movement is increasing. By utilizing its advanced method of virtual simulation technology, the purpose of this paper is to realize the interventional research on the physical education movement and comprehensive health quality in the environment of artificial intelligence. This paper proposes to use the virtual simulation technology and Kinect algorithm in artificial intelligence to design the virtual sports simulation teaching mode. The functional module design part where the Kinect algorithm helps the teaching of virtual sports simulation experiments, which is helpful to analyze and solve the objective system imbalance and ecological imbalance in online physical education teaching. By using the principles and rules of the Mean Shift image segmentation algorithm for reference, the investigation and research on the comprehensive health quality of students are carried out, so as to realize the ecologicalization of the virtual sports school. In the investigation and research on the comprehensive quality of students, the results show that the overall quality of these students who has reached the level of qualified or unqualified is accounting for about 30% of the total number. It is worth noting that in terms of scientific and cultural quality, only 43.34% of all students have excellent grades. It can be seen that the important training goal of current school research is how to use reasonable and effective methods and strategies to improve students' scientific and cultural level, and improve students' other comprehensive scores at the same time.

Evaluation of biventricular longitudinal myocardial function in normal fetuses at different gestational ages using ultrasonic velocity vector imaging.

Objective: This study aimed to evaluate biventricular myocardial function and biventricular longitudinal global myocardial function of fetuses at different gestational weeks using ultrasonic velocity vector imaging. Methods: A total of 127 pregnant women were enrolled and divided into five groups according to the gestational age of their fetuses. The velocity, strain, and strain rate of the left and right ventricles were measured, and these biventricular parameters were compared between the groups. The global parameters of the biventricular myocardium were also compared. Results: A pairwise comparison revealed that the differences in biventricular velocity and strain rate between groups in adjacent gestational weeks were not statistically significant (P > 0.05), but velocity increased with gestational age. A comparison of fetal longitudinal global myocardial parameters revealed that the global velocity, strain, and strain rate of the right ventricle were higher than those of the left ventricle, and the differences were statistically significant (P < 0.05) in all groups. Conclusion: The peak velocities of the fetal left and right ventricles increased with gestational age, but the global strain and strain rate did not, suggesting that fetal myocardial function is mature and constant in the middle and late stages of pregnancy and can more reliably reflect myocardial deformation. The peak systolic velocity, global strain, and peak strain rate of the right ventricle were higher than those of the left ventricle, suggesting that the right ventricle dominates longitudinal systolic movement from the second trimester of pregnancy.

Transcriptomic Analysis Reveals the Positive Role of Abscisic Acid in Endodormancy Maintenance of Leaf Buds of Magnolia wufengensis.

Magnolia wufengensis (Magnoliaceae) is a deciduous landscape species, known for its ornamental value with uniquely shaped and coloured tepals. The species has been introduced to many cities in south China, but low temperatures limit the expansion of this species in cold regions. Bud dormancy is critical for plants to survive in cold environments during the winter. In this study, we performed transcriptomic analysis of leaf buds using RNA sequencing and compared their gene expression during endodormancy, endodormancy release, and ecodormancy. A total of 187,406 unigenes were generated with an average length of 621.82 bp (N50 = 895 bp). In the transcriptomic analysis, differentially expressed genes involved in metabolism and signal transduction of hormones especially abscisic acid (ABA) were substantially annotated during dormancy transition. Our results showed that ABA at a concentration of 100 µM promoted dormancy maintenance in buds of M. wufengensis. Furthermore, the expression of genes related to ABA biosynthesis, catabolism, and signalling pathway was analysed by qPCR. We found that the expression of MwCYP707A-1-2 was consistent with ABA content and the dormancy transition phase, indicating that MwCYP707A-1-2 played a role in endodormancy release. In addition, the upregulation of MwCBF1 during dormancy release highlighted the enhancement of cold resistance. This study provides new insights into the cold tolerance of M. wufengensis in the winter from bud dormancy based on RNA-sequencing and offers fundamental data for further research on breeding improvement of M. wufengensis.

Physiological and transcriptome analysis of Magnolia denudata leaf buds during long-term cold acclimation.

BACKGROUND: Magonlia denudata is an important perennial tree species of the Magnoliaceae family, known for its ornamental value, resistance to smoke pollution and wind, role in air purification, and robust cold tolerance. In this study, a high-throughput transcriptome analysis of leaf buds was performed, and gene expression following artificial acclimation 22 °C, 4 °C and 0 °C, was compared by RNA sequencing. RESULTS: Over 426 million clean reads were produced from three libraries (22 °C, 4 °C and 0 °C). A total of 74,503 non-redundant unigenes were generated, with an average length of 1173.7 bp (N50 = 1548). Based on transcriptional results, 357 and 235 unigenes were identified as being upregulated and downregulated under cold stress conditions, respectively. Differentially expressed genes were annotated using Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes pathway analyses. The transcriptomic analysis focused on carbon metabolism and plant hormone signal transduction associated with cold acclimation. Transcription factors such as those in the basic helix-loop-helix and AP2/ERF families were found to play an important role in M. denudata cold acclimation. CONCLUSION: M. denudata exhibits responses to non-freezing cold temperature (4 °C) to increase its cold tolerance. Cold resistance was further strengthened with cold acclimation under freezing conditions (0 °C). Cold tolerance genes, and cold signaling transcriptional pathways, and potential functional key components for the regulation of the cold response were identified in M. denudata. These results provide a basis for further studies, and the verification of key genes involved in cold acclimation responses in M. denudata lays a foundation for developing breeding programs for Magnoliaceae species.

Correlation analysis between the parameters of contrast-enhanced ultrasonography in evaluating cervical cancer metastasis and expression of E-cadherin.

The study aims to investigate the correlation between the parameters of contrast-enhanced ultrasonography in evaluating cervical cancer metastasis and expression of E-cadherin. All 120 patients with cervical cancer underwent contrast-enhanced ultrasonography. According to the results of postoperative pathological examination, patients were divided into distant metastasis group (group A), lymph node metastasis without distant metastasis group (group B) and no metastasis group (group C). Expression of E-cadherin in cervical cancer tissues was detected by enzyme-linked immunosorbent assay (ELISA). Correlations between the parameters of contrast-enhanced ultrasonography in evaluating cervical cancer metastasis and expression of E-cadherin were analyzed by Pearsons correlation analysis. Comparison of parameters of contrast-enhanced ultrasonography showed that, the baseline intensity of group A was 11.9±2.2 dB, which was significantly lower than that of group B and C. Baseline intensity of group B was significantly lower than that of group C (13.0±2.4 vs. 15.3±3.6 dB), significant differences were found among three groups (P<0.05). The sensitivity and specificity of the use of enhanced intensity ~83.7 dB in evaluating tumor metastasis of patients with cervical cancer were 82.42 and 79.32%, respectively. Expression level of E-cadherin protein in group A was 0.030±0.003 ng/ml, which was significantly lower than that in group B and C (P<0.05), expression level of E-cadherin protein in group A was significantly lower than that in group C (0.037±0.007 vs. 0.045±0.012 ng/ml), significant differences in the expression level of E-cadherin protein were found among the three groups of cervical cancer patients (P<0.05). Pearsons correlation analysis showed that there was a positive correlation between the baseline intensity of contrast-enhanced ultrasonography and the expression level of E-cadherin (P<0.05), while there was a negative correlation between the enhanced intensity of contrast-enhanced ultrasonography and the expression level of E-cadherin (P<0.05). Contrast-enhanced ultrasonography can be used to determine the tumor metastasis of cervical cancer patients, in addition, the combined use of contrast-enhanced ultrasonography and E-cadherin protein expression can significantly improve the diagnosis and treatment of cervical cancer.

Performance of wastewater subsurface drip emitters at low and normal pressure.

Subsurface drip distribution is an important on-site wastewater treatment technique widely used with various soil types and restricted site conditions. This study evaluated the performance of five subsurface wastewater drip products under eight pressures, ranging from 0 to 310 kPa. Results showed that Netafim Bioline pressure-compensating emitters (Netafim Irrigation Inc., Fresno, California) had an application uniformity coefficient of 95% and a coefficient of variance (C(v)) of 4.9%. The average uniformity coefficient of Geoflow Wasteflow products (Geoflow USA, Charlotte, North Carolina) was 94.4%, with a C(v) value of 6.8%. Flowrate and pressure relationships were developed by analyzing low and normal operational pressure ranges, and R-square values ranged from 1.000 to 0.301. Geoflow pressure-compensating products were non-pressure-compensating emitters under low pressure. Netafim pressure-compensating emitters were partially pressure-compensating under low pressures. In normal operational pressure ranges, both Geoflow and Netafim products were fully pressure-compensating. Netafim pressure-compensating products were characterized as pressure-compensating over the full range of operational pressures.

Uniformity of domestic wastewater effluent application in a subsurface drip distribution system.

An on-site wastewater treatment project with two separate drip fields was operated for 6 years and received no maintenance. The two drip fields (with different design configurations) contained pressure-compensating emitters (PC) and non-pressure-compensating emitters (NPC), respectively, and received wastewater with an average 5-day biochemical oxygen demand concentration of 23 mg/L. Flowrates of the PC emitters reduced from rated average of 3.50 to 1.00 L/h, and the average flowrate of the NPC emitters reduced from 2.00 to 1.53 L/h. The statistical uniformities were 48 and 71%, and the uniformity coefficients were 70 and 86% for PC and NPC emitters, respectively. Significant, but incomplete, recovery was achieved with field-flushing and consecutive shock-chlorination treatments of 500 and 1000 mg/L.