Vernalization promotes bolting in sugar beet by inhibiting the transcriptional repressors of BvGI.

Sugar beet (Beta vulgaris L.), a biennial sugar crop, contributes about 16% of the world's sugar production. The transition from vegetative growth, during which sugar accumulated in beet, to reproductive growth, during which sugar exhausted in beet, is determined by vernalization and photoperiod. GIGANTEA (GI) is a key photoperiodic flowering gene that is induced by vernalization in sugar beet. To identify the upstream regulatory factors of BvGI, candidate transcription factors (TF) that were co-expressed with BvGI and could bind to the BvGI promoter were screened based on weighted gene co-expression network analysis (WGCNA) and TF binding site prediction. Subsequently, their transcriptional regulatory role on the BvGI was validated through subcellular localization, dual-luciferase assays and yeast transformation tests. A total of 7,586 differentially expressed genes were identified after vernalization and divided into 18 co-expression modules by WGCNA, of which one (MEcyan) and two (MEdarkorange2 and MEmidnightblue) modules were positively and negatively correlated with the expression of BvGI, respectively. TF binding site predictions using PlantTFDB enabled the screening of BvLHY, BvTCP4 and BvCRF4 as candidate TFs that negatively regulated the expression of BvGI by affecting its transcription. Subcellular localization showed that BvLHY, BvTCP4 and BvCRF4 were localized to the nucleus. The results of dual-luciferase assays and yeast transformation tests showed that the relative luciferase activity and expression of HIS3 was reduced in the BvLHY, BvTCP4 and BvCRF4 transformants, which suggested that the three TFs inhibited the BvGI promoter. In addition, real-time quantitative reverse transcription PCR showed that BvLHY and BvTCP4 exhibited rhythmic expression characteristics similar to that of BvGI, while BvCRF4 did not. Our results revealed that vernalization crosstalked with the photoperiod pathway to initiate bolting in sugar beet by inhibiting the transcriptional repressors of BvGI.

Outcomes of patients in nasopharyngeal adenoid cystic carcinoma in the IMRT era: a single-center experience.

OBJECTIVE: Nasopharyngeal adenoid cystic carcinoma (NACC) is a rare malignancy with special biological features. Controversies exist regarding the treatment approach and prognostic factors in the IMRT era. This study aimed to evaluate the long-term outcomes and management approaches in NACC. METHODS: Fifty patients with NACC at our institution between 2010 and 2020 were reviewed. Sixteen patients received primary radiotherapy (RT), and 34 patients underwent primary surgery. RESULTS: Between January 2010 and October 2020, a total of 50 patients with pathologically proven NACC were included in our analysis. The median follow-up time was 58.5 months (range: 6.0-151.0 months). The 5-year overall survival rate (OS) and progression-free survival rate (PFS) were 83.9% and 67.5%, respectively. The 5-year OS rates of patients whose primary treatment was surgery and RT were 90.0% and 67.3%, respectively (log-rank P = 0.028). The 5-year PFS rates of patients whose primary treatment was surgery or RT were 80.8% and 40.7%, respectively (log-rank P = 0.024). Multivariate analyses showed that nerve invasion and the pattern of primary treatment were independent factors associated with PFS. CONCLUSIONS: Due to the relative insensitivity to radiation, primary surgery seemed to provide a better chance of disease control and improved survival in NACC. Meanwhile, postoperative radiotherapy should be performed for advanced stage or residual tumours. Cranial nerve invasion and treatment pattern might be important factors affecting the prognosis of patients with NACC.

Comparative proteomic analysis on chloroplast proteins provides new insights into the effects of low temperature in sugar beet.

BACKGROUND: Low temperature, which is one of the main environmental factors that limits geographical distribution and sucrose yield, is a common abiotic stress during the growth and development of sugar beet. As a regulatory hub of plant response to abiotic stress, activity in the chloroplasts is related to many molecular and physiological processes, particularly in response to low temperature stress. RESULTS: The contents of chlorophyll (Chl) and malondialdehyde (MDA), relative electrical conductivity (REL), and superoxide dismutase (SOD) activity were measured. The results showed that sugar beet could manage low temperature stress by regulating the levels of Chl, REL and MDA, and the activity of SOD. The physiological responses indicated that sugar beets respond positively to low temperature treatments and are not significantly damaged. Moreover, to determine the precise time to response low temperature in sugar beet, well-known abiotic stresses-responsive transcript factor family, namely DEHYDRATION RESPONSIVE ELEMENT BINDING PROTEIN (DREB), was selected as the marker gene. The results of phylogenetic analyses showed that BvDREBA1 and BvDREBA4 were in the same branch as the cold- and drought-responsive AtDREB gene. In addition, the expression of BvDREBs reached its maximum level at 24 h after low temperature by RNA-Seq and qRT-PCR analysis. Furthermore, the changes in chloroplast proteome after low temperature at 24 h were detected using a label-free technique. A total of 416 differentially expressed proteins were identified. GO enrichment analysis showed that 16 GO terms were significantly enriched, particularly chloroplast stroma, chloroplast envelope, and chloroplast thylakoid membrane. It is notable that the transport of photosynthetic proteins (BvLTD and BvTOC100), the formation of starch granules (BvPU1, BvISA3, and BvGWD3) and the scavenging of reactive oxygen species (BvCu/Zn-SOD, BvCAT, BvPrx, and BvTrx) were the pathways used by sugar beets to respond to low temperatures at an early stage. CONCLUSIONS: These results provide a preliminarily analysis of how chloroplasts of sugar beet respond to low temperature stress at the translational level and provide a theoretical basis for breeding low temperature resistant varieties of sugar beet.

A Chromosome-Scale Genome Assembly of Paper Mulberry (Broussonetia papyrifera) Provides New Insights into Its Forage and Papermaking Usage.

Paper mulberry (Broussonetia papyrifera) is a well-known woody tree historically used for Cai Lun papermaking, one of the four great inventions of ancient China. More recently, Paper mulberry has also been used as forage to address the shortage of feedstuff because of its digestible crude fiber and high protein contents. In this study, we obtained a chromosome-scale genome assembly for Paper mulberry using integrated approaches, including Illumina and PacBio sequencing platform as well as Hi-C, optical, and genetic maps. The assembled Paper mulberry genome consists of 386.83 Mb, which is close to the estimated size, and 99.25% (383.93 Mb) of the assembly was assigned to 13 pseudochromosomes. Comparative genomic analysis revealed the expansion and contraction in the flavonoid and lignin biosynthetic gene families, respectively, accounting for the enhanced flavonoid and decreased lignin biosynthesis in Paper mulberry. Moreover, the increased ratio of syringyl-lignin to guaiacyl-lignin in Paper mulberry underscores its suitability for use in medicine, forage, papermaking, and barkcloth making. We also identified the root-associated microbiota of Paper mulberry and found that Pseudomonas and Rhizobia were enriched in its roots and may provide the source of nitrogen for its stems and leaves via symbiotic nitrogen fixation. Collectively, these results suggest that Paper mulberry might have undergone adaptive evolution and recruited nitrogen-fixing microbes to promote growth by enhancing flavonoid production and altering lignin monomer composition. Our study provides significant insights into genetic basis of the usefulness of Paper mulberry in papermaking and barkcloth making, and as forage. These insights will facilitate further domestication and selection as well as industrial utilization of Paper mulberry worldwide.

Electrospun LiFePO4/C Composite Fiber Membrane as a Binder-Free, Self-Standing Cathode for Power Lithium-Ion Battery.

A LiFePO4/C composite fiber membrane was fabricated by the electrospinning method and subsequent thermal treatment. The thermal decomposition process was analyzed by TG/DSC, the morphology, microstructure and composition were studied using SEM, TEM, XRD, Raman, respectively. The results indicated that the prepared LiFePO4/C composite fibers were composed of nanosized LiFePO4 crystals and amorphous carbon coatings, which formed a three dimensional (3D) long-range networks, greatly enhanced the electronic conductivity of LiFePO4 electrode up to 3.59× 10-2 S · cm-2. The 3D LiFePO4/C fiber membrane could be directly used as a binder-free, self-standing cathode for lithium-ion battery, and exhibited an improved capacity and rate performance. The LiFePO4/C composite electrode delivered a discharge capacity of 116 mAh·g-1, 109 mAh·g-1, 103 mAh·g-1, 91 mAh·g-1, 80 mAh·g-1 at 0.1 C, 0.5 C, 1 C, 3 C, 5 C, respectively. And a stable cycling performance was also achieved that the specific capacity could retain 75 mA·g-1 after 500 cycles at 5 C. Therefore, this LiFePO4/C composite fiber membrane was promising to be used as a cathode for power lithium ion battery.

Phosphoproteomic Analysis of Paper Mulberry Reveals Phosphorylation Functions in Chilling Tolerance.

Paper mulberry is a valuable woody species with a good chilling tolerance. In this study, phosphoproteomic analysis, physiological measurement, and mRNA quantification were employed to explore the molecular mechanism of chilling (4 °C) tolerance in paper mulberry. After chilling for 6 h, 427 significantly changed phosphoproteins were detected in paper mulberry seedlings without obvious physiological injury. When obvious physiological injury occurred after chilling for 48 h, a total of 611 phosphoproteins were found to be significantly changed at the phosphorylation level. Several protein kinases, especially CKII, were possibly responsible for these changes according to conserved sequence analysis. The results of Gene Ontology analysis showed that phosphoproteins were mainly responsible for signal transduction, protein modification, and translation during chilling. Additionally, transport and cellular component organization were enriched after chilling for 6 and 48 h, respectively. On the basis of the protein-protein interaction network analysis, a protein kinase and phosphatases hub protein (P1959) were found to be involved in cross-talk between Ca2+, BR, ABA, and ethylene-mediated signaling pathways. We also highlighted the phosphorylation of BpSIZ1 and BpICE1 possibly impacted on the CBF/DREB-responsive pathway. From these results, we developed a schematic for the chilling tolerance mechanism at phosphorylation level.

Proteomic changes induced by potassium deficiency and potassium substitution by sodium in sugar beet.

In this study, sugar beets (Beta vulgaris L.) were grown at different K(+)/Na(+) concentrations: mmol/L, 3/0 (control); 0.03/2.97 (K-Na replacement group; T(rep)); 0.03/0 (K deficiency group; T(def)) in order to investigate the effects of K(+) deficiency and replacement of K(+) by Na(+) on plant proteomics, and to explore the physiological processes influenced by Na(+) to compensate for a lack of K(+). After 22 days, fresh and dry weight as well as the Na(+) and K(+) concentration were measured and changes in proteomics were tested by 2D gel electrophoresis. Interestingly, Na(+) showed stimulation in growth of seedlings and hindrance of K(+) assimilation in T(rep). Significant changes were also observed in 27 protein spots among the treatments. These are proteins involved in photosynthesis, cellular respiration, protein folding and degradation, stress and defense, other metabolisms, transcription related, and protein synthesis. A wide range of physiological processes, including light reaction, CO2 assimilation, glycolysis, and tricaboxylic acid cycle, was impaired owing to K(+) starvation. Compensating for the effect of K(+) starvation, an increase in photosynthesis was also observed in T(rep). However, we also found a limitation of cellular respiration by Na(+). Na(+) is therefore in some ways able to recover damage due to K deficiency at protein level, but cannot functionally replace K as an essential nutrient.

Inhibition effect of Peg-IFNα-2b and Imatinib alone or combination on imatinib-resistant gastrointestinal stromal tumors cell lines.

To investigate the inhibition effect of polyethylene glycol interferon α-2b and imatinib alone or combination on imatinib-resistant GIST cell lines, and to explore the possible mechanism. Imatinib resistant GIST cell lines (GIST-R) were exposured to either Peg-IFNα-2b or imatinib alone or combination. The proliferative inhibition rates and the combination index (CI) values of GIST-R cells were detected by MTT assay. The apoptotic rates of GIST-R cells were detected by flow cytometry. The expression levels of phospho-mammalian target of rapamycin (p-mTOR), and Bcl-2 of GIST-R cells were analyzed by Western blot. GIST-R cells presents remarkable resistance to imatinib, and the resistance index (RI) were (P<0.05). And The proliferative inhibition rate and the apoptotic rate of GIST-R cells in combination of Peg-IFNα2b and Imatinib group were higher than those in Peg-IFNα-2b or imatinib alone group (P<0.05). The CI value of Peg-IFNα-2b and imatinib was less than each alone, which had a synergistic effect (CI=0.63). As compared with the control (GIST-R cells without any treatment), the expression levels of p-mTOR and Bcl-2 proteins of GIST-R cells in combination of Peg-IFNα-2b and imatinib group were decreased (P<0.01). The combination of Peg-IFNα-2b and imatinib generats a synergistic effect in GIST-R cells, and reversal of drug resistance. This effect may be related with apoptosis and down-regulation of the expression of p-mTOR.

Effect of berberine on acetylcholine-induced atrial fibrillation in rabbit.

The purpose of this study was to test the efficacy of Berberine (Ber) on atrial fibrillation (AF) induced by acetylcholine (ACh) and explore its underlying mechanisms of action. In vivo electrophysiology experiments were performed in adult anesthetized rabbits. Single atrial myocytes were isolated from rabbit hearts and action potentials recorded using patch clamp techniques. AF was induced by rapid atrial burst pacing during intravenous (IV) ACh infusion alone or with IV Ber. Compared to the Baseline, IV Ber (2 mg/kg) prolonged the RR interval and effective refractory period (195 ± 10 vs. 215 ± 11 msec; 80 ± 4 vs. 85 ± 5 msec, respectively; both P<0.05). The induced rate of sustained 1 min AF was lower during ACh infusion with Ber than during ACh infusion alone (4/10 vs. 30/35, P<0.01). The termination rate of ACh-induced AF was higher with IV Ber (1 mg/kg) than with IV saline (sustained 1 min AF: 6/8 vs. 6/20, sustained 10 min AF: 8/10 vs. 1/6, both P<0.05). ACh perfusion significantly shortened the action potential duration (APD) of isolated atrial myocytes (APD50: 152 ± 13 vs. 81 ± 10 msec; APD90: 256 ± 19 vs. 132 ± 13 msec, both P<0.01). Application of Ber reversed the APD shortening induced by ACh (APD50: 81 ± 10 vs. 134 ± 15 msec; APD90: 132 ± 13 vs: 213 ± 17 msec, both P<0.01). We conclude that Ber suppresses ACh-induced AF in the rabbit by increasing atrial effective refractory period and prolonging the APD of atrial myocytes.

Preparation and Properties of Double-Sided AgNWs/PVC/AgNWs Flexible Transparent Conductive Film by Dip-Coating Process.

The double-sided transparent conductive films of AgNWs/PVC/AgNWs using the silver nanowires and PVC substrate were fabricated by the dip-coating process followed by mechanical press treatment. The morphological and structural characteristics were investigated by scanning electron microscope (SEM) and atomic force microscope (AFM), the photoelectric properties and mechanical stability were measured by ultraviolet-visible spectroscopy (UV-vis) spectrophotometer, four-point probe technique, 3M sticky tape test, and cyclic bending test. The results indicate that the structure and photoelectric performances of the AgNWs films were mainly affected by the dipping and lifting speeds. At the optimized dipping speed of 50 mm/min and lifting speed of 100 mm/min, the AgNWs are evenly distributed on the surface of the PVC substrate, and the sheet resistance of AgNWs film on both sides of PVC is about 60 Ω/sq, and the optical transmittance is 84.55 % with the figure of merit value up to 35.8. The film treated with the 10 MPa pressure shows excellent adhesion and low surface roughness of 17.8 nm and maintains its conductivity with the sheet resistance change of 17 % over 10,000 cyclic bends.

Transcriptional regulation of the paper mulberry under cold stress as revealed by a comprehensive analysis of transcription factors.

BACKGROUND: Several studies have focused on cold tolerance in multiple regulated levels. However, a genome-scale molecular analysis of the regulated network under the control of transcription factors (TFs) is still lacking, especially for trees. To comprehensively identify the TFs that regulate cold stress response in the paper mulberry and understand their regulatory interactions, transcriptomic data was used to assess changes in gene expression induced by exposure to cold. RESULTS: Results indicated that 794 TFs, belonging to 47 families and comprising more than 59% of the total TFs of this plant, were involved in the cold stress response. They were clustered into three groups, namely early, intermediate and late responsive groups which contained 95, 550 and 149 TFs, respectively. Among of these differentially expressed TFs, one bHLH, two ERFs and three CAMTAs were considered to be the key TFs functioning in the primary signal transduction. After that, at the intermediate stage of cold stress, there were mainly two biological processes that were regulated by TFs, namely cold stress resistance (including 5 bHLH, 14 ERFs, one HSF, 4 MYBs, 3 NACs, 11 WRKYs and so on) and growth and development of lateral organ or apical meristem (including ARR-B, B3, 5 bHLHs, 2 C2H2, 4 CO-like, 2 ERF, 3 HD-ZIP, 3 YABBYs, G2-like, GATA, GRAS and TCP). In late responsive group, 3 ARR-B, C3H, 6 CO-like, 2 G2-like, 2 HSFs, 2 NACs and TCP. Most of them presented the up-regulated expression at 12 or 24 hours after cold stress implied their important roles for the new growth homeostasis under cold stress. CONCLUSIONS: Our study identified the key TFs that function in the regulatory cascades mediating the activation of downstream genes during cold tress tolerance in the paper mulberry. Based on the analysis, we found that the AP2/ERF, bHLH, MYB, NAC and WRKY families might play the central and significant roles during cold stress response in the paper mulberry just as in other species. Meanwhile, many other TF families previously reported as involving in regulation of growth and development, including ARF, DBB, G2-like, GRF, GRAS, LBD, WOX and YAABY exhibited their important potential function in growth regulation under cold stress.

[Effect of hydroxyethyl starch 130/0.4 on S100B protein level and cerebral oxygen metabolism in open cardiac surgery under cardiopulmonary bypass].

OBJECTIVE: To observe the effect of hydroxyethyl starch (HES) 130/0.4 on S100B protein level and cerebral metabolism of oxygen in open cardiac surgery under cardiopulmonary bypass (CPB) and to explore whether it has the protective effect of 6%HES130/0.4 as priming solution on cerebral injury during CPB and explore the probable mechanism. METHODS: Forty patients with atrioseptal defect or ventricular septal defect scheduled for elective surgical repair under CPB with moderate hypothermia were randomly divided into two equal groups: HES 130/0.4 group (HES group) in which HES 130/0.4 (voluven) was used as priming solution and gelatin group (GRL group) in which gelofusine (succinylated gelatin) was used as priming solution. ECG, heart rate (HR), blood pressure (BP), mean arterial pressure (MAP), central venous pressure (CVP), arterial partial pressure of oxygen (P(a)O(2),), arterial partial pressure of carbon dioxide (P(et)CO(2)) and body temperature (naso-pharyngeal and rectal) were continuously monitored during the operation. Blood samples were obtained from the central vein for determination of blood concentrations of S100B protein at the following time points: before CPB (T(0)), 20 minutes after the beginning of CPB (T(1)), immediately after the termination of CPB (T(2)), 60 minutes after the termination of CPB (T(3)), and 24 hours after the termination of CPB (T(4)). The serum S100B protein levels were measured by ELISA. At the same time points blood samples were obtained from the jugular vein and radial artery to undergo blood gas analysis and measurement of blood glucose, based on which the cerebral oxygen metabolic rate/cerebral metabolic rate of glucose (CMRO(2)/CMR(GLU)) was calculated. RESULTS: Compared with the time point of immediately before CPB (T(0)), The S100B protein level of the 2 groups began to increase since the time point T(1), peaked at the time point T(2), began to decrease gradually since the time point T(3), and were still significantly higher than those before CPB at the time point T(4) (all P < 0.01), and the S100B protein levels at different time points of the HES group were all significantly lower than those of the GEL group (all P < 0.01). The S(jv)O(2) and CMRO(2)/CMR(GLU) levels of both groups increased at the time point T(1), decreased at the time points T(2) and T(3), and then restored to normal at the time points T(4). In the GEL group there were no significant differences in the levels between any 2 different time points, however, in the HES group S(jv)O(2) and CMRO(2)/CMR(GLU) levels at T(1) was significantly higher than those at the other time points (P < 0.05 or P < 0.01). CONCLUSION: S100B protein increases significantly in open cardiac surgery under CPB. HES130/0.4 lowers the S100B protein levels from the beginning of CPB to one hour after the termination of CPB with the probable mechanism of improving the cerebral metabolism of oxygen. 6%HES130/0.4 as priming solution may play a protective role in reduction of cerebral injury during CPB and open cardiac surgery.

[Relevant factors of lymphatic metastasis in rectal carcinoma].

OBJECTIVE: To investigate the rules of lymphatic metastasis of rectal carcinoma, and to help clinical diagnosis and treatment. METHODS: A retrospective analysis was performed in the 979 patients with rectal carcinoma who underwent surgical resection from 1995 to 2004. The associations between lymphatic metastasis and clinicopathologic variables were evaluated by Chi-squared test and logistic regression. RESULTS: The rate of lymph node metstasis was 71.4% for patients younger than 30 years old, 40.7% in the patients with tumor diameters over 6 centimeters, 82.5% in the patients with extraneous tumor invasion, 71.6% for patients of poor-differentiated adenocarcinoma, 70.4% for patients with mucoid adenocarcinoma, 100% for patients with signet-ring cell carcinoma and 46.4% for patients with more than half intestinal circumference invasion. Logistic regression analysis showed that the degree of lymphatic metastasis was related to the differentiating degrees, depths of tumor invasion and intestinal circumference invasion, and the differentiating degree was the major factor. CONCLUSION: The lymphatic metastasis of rectal carcinoma is related to age, tumor size, intestinal circumference invasion, depth of tumor invasion and the differentiating degree of the tumor; the differentiating degree is the major factor.

Radioimmunoimaging of colorectal cancer using (99m)Tc labeled monoclonal antibody.

AIM:To determine whether Hb3 and its fragment F(ab')(2) have practical value in radioimmunoimaging of colorectal cancer.METHODS:Intact Hb3 was purified by hydroxylapatite chromatography.The fragment F(ab') (2) was prepared by cold digestion and purified as intact Hb3.Hb3 and its fragment F(ab') (2) were labeled with 99mTc by direct labeling method using SnCl(2) as reducing agent. The radioactive doses ranged from 15 to 40 mCi.The imaging was accomplished by single photon emission computered tomograph (SPECT) with imaging time ranging from 2.5 to 48 hours. In this study, 10 patients were selected. Among them, 7 were administered with intact Hb3, and 3 with F(ab') (2) fragment. All the patients were diagnosed as having colorectal adenocarcinoma.RESULTS:After purification, intact Hb3 and its fragment F(ab') (2) were fit for radioimmunoimaging. The percentage of labeling of (99m)Tc to Hb3 or F(ab') (2) was 80.6%-91.5%. Among the 10 patients, 3 of 7 patients administered with intact Hb3 had positive scans, the other 4 had negative scans, and 2 of 3 patients administered with F(ab') (2)had positive scans, the other 1 had negative scans.CONCLUSION:The results showed that both intact Hb3 and its F(ab') (2) have some practical value in radioimmunoimaging of colorectal cancer, and the effects of imaging with F(ab') (2) was better than that with intact Hb3.