Genome-wide identification of GhRLCK-VII subfamily genes in Gossypium hirsutum and investigation of their functions in resistance to Verticillium wilt.

BACKGROUND: The receptor-like cytoplasmic kinases subfamily VII (RLCK-VII) is critical in regulating plant growth, development, and pattern-triggered immunity. However, a comprehensive exploration of these genes in the allotetraploid Gossypium hirsutum is still lacking. This study aimed to identify RLCK-VII genes in G. hirsutum and investigate their evolutionary history, structural features, expression patterns, and role in plant defense. RESULTS: Seventy-two RLCK-VII genes in the G. hirsutum genome were unveiled and classified into nine groups following their phylogenetic analysis with Arabidopsis thaliana. Group VII-1 was the largest, accounting for 28%, while Groups VII-2 and VII-3 had only one member each. The analysis using MCScanX revealed that these 72 genes formed 166 collinear gene pairs and were resided on 26 chromosomes of G. hirsutum, suggesting that they were derived from whole genome segmental duplication events. Their calculated Ka/Ks values were below one, implying the occurrence of purification selection during the evolution and inhibition of gene function differentiation/loss. All members of the RLCK-VII subfamily possessed two conserved domains, PKinase-Tyr and PKinase, and several conserved PBS1 kinase subdomains, individually included in one of the ten motifs identified using MEME. The RNA-Seq results showed that RLCK-VII genes exhibited different spatiotemporal expression, indicating their involvement in cotton growth, development, and defense responses to Verticillium dahliae. The transcription patterns of RLCK-VII genes found by RNA-Seq were further validated using qRT-PCR assays after inoculating "20B12" (cotton cultivar) with "V991" (V. dahliae). The virus-induced gene silencing (VIGS) assays uncovered that two RLCK-VII genes (Gohir.A13G227248 and Gohir.A10G219900) were essential to G. hirsutum resistance to Verticillium wilt. CONCLUSIONS: These observations offer valuable insight into the attributes and roles of RLCK-VII genes in G. hirsutum, potentially enable the breeding of new cotton cultivars with enhanced resistance to Verticillium wilt.

Two New Red/Near-Infrared Ir(â ¢) Complexes with Reversible and Force-Induced Enhanced Mechanoluminescence.

Two novel ionic red/near-infrared Ir(III) complexes (Ir1 and Ir2) were reasonably designed and prepared using 2-(1-isoquinolinyl)-9,10-anthraquinone as the main ligand and 4,4'-dimethyl-2,2'-bipyridyl and 4,4'-dimethoxy-2,2'-bipyridyl as the auxiliary ligands, respectively. Both complexes showed bright phosphorescence in solution (peak at 618 nm with a shoulder at 670 nm). Interestingly, the phosphorescence peak of two Ir(III) complexes showed a blue-shift of about 36 nm after being ground. Simultaneously, both complexes exhibited mechanical force-induced enhanced emission, and the intensity of the luminescence for Ir1 and Ir2 increased by around two times compared to the one before being ground, respectively. Powder X-ray diffraction (PXRD) and time-dependent density functional theory (TD-DFT) calculation were utilized to understand well the mechanism of this phenomenon and suggested that the destruction of the well-ordered crystalline nature and the decline in triplet-triplet annihilation maybe responsible for the pressure-induced blue-shift and the enhancement of the phosphorescence.

Two Novel Neutral Cyclometalated Iridium(III) Complexes Based on 10,11,12,13-Tetrahydrodibenzo[a,c]phenazine for Efficient Red Electroluminescence.

Two novel neutral phosphorescent iridium(III) complexes (Ir1 and Ir2) were rationally designed and synthesized with high yields using 10,11,12,13-tetrahydrodibenzo[a,c]phenazine as the main ligand. The two complexes showed bright-red phosphorescence (625 nm for Ir1, and 620 nm for Ir2, in CH2Cl2), high-luminescence quantum efficiency (0.32 for Ir1, and 0.35 for Ir2), obvious solvatochromism and good thermostability. Then, they were used to fabricate high-efficiency red OLEDs via vacuum evaporation; the maximum current efficiency, power efficiency, and external quantum efficiency of the red devices based on Ir1 and Ir2 are 13.47/15.22 cd/A, 10.35/12.26 lm/W, and 10.08/7.48%, respectively.

Evaluation and validation of reference genes in Cymbidium faberi for real-time quantitative PCR.

For all organs at all Cymbidium faberi stages, ACT, UBQ3 and GAPDH can be selected as reference genes. For organs of the vegetative stage, UBQ2 and UBQ3 can be chosen for analysis of normalized gene expression. For the bud stage, ACT and UBQ3 can be used for analysis of gene expression. For the full blossom stage, ACT, UBQ3 and UBQ2 can be introduced into relative gene expression analysis. For vegetative organs, UBQ2 and ACT can be used as reference genes. For reproductive organs, ACT, UBQ3 and UBQ2 can be used as a reference for data processing. CfAG1 gene expression is more consistent when UBQ3, GAPDH and ACT are used as reference genes.

Comparative Transcriptome Analysis Reveals Genes Associated with the Gossypol Synthesis and Gland Morphogenesis in Gossypium hirsutum.

Gossypium hirsutum is an important source of natural textile fibers. Gossypol, which is a sesquiterpenoid compound mainly existing in the cotton pigment glands, can facilitate resistance to the stress from diseases and pests. The level of gossypol in the cotton is positively correlated to the quantity of pigment glands. However, the underlying regulatory mechanisms of gossypol synthesis and gland morphogenesis are still poorly understood, especially from a transcriptional perspective. The transcripts of young leaves and ovules at 30 DPA of the glanded plants and glandless plants were studied by RNA-Seq and 865 million clean reads were obtained. A total of 34,426 differentially expressed genes (DEGs) were identified through comparative transcriptome analysis. Genes related to gossypol synthesis or gland morphogenesis displayed significant differential expression between the two cultivars. Functional annotation revealed that the candidate genes related to catalytic activity, the biosynthesis of secondary metabolites, and biomolecular decomposition processes. Our work herein unveiled several potential candidate genes related to gossypol synthesis or gland morphogenesis and may provide useful clues for a breeding program of cotton cultivars with low cottonseed gossypol contents.

[Cloning and expression analysis of 8 bHLH transcription factors in Panax quinquefolius].

A total of 8 bHLH transcription factors were cloned from Panax quinquefolius and the response of them to methyl jasmonate(MeJA) was studied.To be specific, based on the preliminary transcriptome screening, 8 bHLH transcription factors were cloned with seedlings which had been cultured for 3 weeks.The content of ginsenosides Rg_1, Re, and Rb_1, and total saponins in the adventitious roots of P.quinquefolius was determined at different time of MeJA treatment by high performance liquid chromatography(HPLC) and spectrophotometry.Real-time quantitative polymerase chain reaction(PCR) was used to detect the relative expression of 8 transcription factors after MeJA treatment.The correlation between the relative expression of the 8 transcription factors and the saponin content after MeJA treatment was checked by Pearson's correlation analysis.The results showed that the PCR products(Pq-bHLH21-Pq-bHLH28) of the 8 bHLH transcription factors were 762-2 013 bp in length.They were submitted to NCBI to obtain the Genbank access numbers.The proteins yielded from Pq-bHLH21-Pq-bHLH28 showed amino acid sequence identity of 24.90%, and each amino acid sequence had the bHLH(Basic Helix-loop-helix) conserved domain and belonged to the bHLH family.The 5 amino acid sequences of Pq-bHLH22 and Pq-bHLH24-Pq-bHLH27 contained the bHLH-MYC N domain, which belonged to the MYC transcription factors.Pq-bHLH21-Pq-bHLH28 responded to MeJA within 48 h of treatment.At 72 h, the expression of Pq-bHLH24 reached 106.53 folds the highest in the treatment group.Pq-bHLH25, Pq-bHLH27, and Pq-bHLH28 showed synergic expression.Pq-bHLH21 may re-gulate the biosynthetic pathway of ginsenoside Rb_1, while Pq-bHLH22, Pq-bHLH25, and Pq-bHLH28 were in significantly positive correlation with the biosynthetic pathway of ginsenoside Re.The result lays a foundation for further verifying the regulation of ginsenoside biosynthesis by bHLH transcription factors.

Solution-Processed Yellow Organic Light-Emitting Diodes Based on Two New Ionic Ir (III) Complexes.

Two new and efficient cationic yellow-emissive Ir (III) complexes (Ir1 and Ir2) are rationally designed by using 2-(4-chloro-3-(trifluoromethyl)phenyl)-4-methylquinoline as the main ligand, and, respectively, 4,4'-dimethyl-2,2'-bipyridyl and 4,4'-dimethoxy-2,2'-bipyridyl as the ancillary ligands. Both complexes show enhanced phosphorescence (546 nm with 572 nm as shoulder and high phosphorescent quantum efficiency in solution, which is in favor of efficient solution-processed phosphorescent organic light-emitting diodes. Compared with Ir2, the Ir1-based device displays excellent device performance, with maximum external quantum efficiency, current efficiency, and power efficiency of up to 7.92%, 26.32 cd/A and 15.31 lm/W, respectively, thus proving that the two new ionic Ir (III) complexes exhibit great potential for future solution-processed electroluminescence.

Selection and validation of reference genes for quantitative real-time PCR in Cymbidium sinense.

Selection of reference genes (RGs) is important for the accurate analysis of real-time quantitative PCR (RT-qPCR) results. This study screened RGs of Cymbidium sinense for more accurate quantification of target genes. The two most stable RGs for all tissues were ACT and EF1α, those for vegetative organs were UBQ3 and ACT, while those for reproductive organs as well as organs in the full flowering stage were EF1α and ACT. The AGAMOUS (CsAG1) expression level was verified using EF1α, ACT, GAPDH, UBQ2 and UBQ3 as RG. The expression profile of CsAG1 was consistent when normalized with EF1α, ACT and UBQ3. The results have practical value for the expression of key genes during the development of C. sinense.

Folic Acid Attenuates High-Fat Diet-Induced Osteoporosis Through the AMPK Signaling Pathway.

Objective: Obesity caused by a high-fat diet (HFD) will expand adipose tissue and cause chronic low-grade systemic inflammation, leading to osteoporosis. Folic acid (FA) is a water-soluble vitamin that plays an essential role in regulating blood lipids and antioxidants. However, the effects and underlying mechanisms of FA in osteoporosis induced by an HFD remain poorly understood. This study aimed to investigate the effect of FA on bone health by using HFD-induced osteoporosis mice. Materials and Methods: Mice were fed a normal diet, HFD or an HFD supplemented with FA (20 µg/ml in drinking water) for 16 weeks. Throughout the 16 weeks study period, the rats were weighed once every week. GTT, ITT and lipid indexes were detected to evaluate the effects of FA on lipid metabolism in the HFD-fed mice. Morphological and structural changes of the femur and tibial bone were observed using micro-CT, HE staining and bone conversion parameters. The expression of MDA, SOD and inflammatory factors were detected to evaluate the effects of FA on oxidative stress and inflammatory response in the HFD-fed mice. Quantitative real-time PCR and Western blot (WB) were used to investigate the AMPK signaling pathway. Results: After the intervention of FA, the body fat rate of obese mice was reduced, and related metabolic disorders such as insulin resistance, hyperlipidemia, and systemic inflammation were alleviated. In correlation with those modifications, FA attenuated bone loss and improved bone microarchitecture, accompanied the number of osteoclasts and adipocytes decreased. Furthermore, FA promoted the phosphorylation of AMPK, thereby promoting the expression of Carnitine palmitoyltransferase 1 (CPT1), nuclear factor erythroid-2 related factor 2 (Nrf2) and antioxidant enzymes. Conclusion: These findings suggest that FA may modulate lipid metabolism and oxidative stress responses activating the AMPK signaling pathway, thereby alleviating HFD-induced osteoporosis. The results from our study provide experimental evidence to prevent HFD-induced osteoporosis.

Study on asymmetric diffraction of acoustic parity-time-symmetric gratings using rigorous coupled-wave analysis.

The method of rigorous coupled-wave analysis (RCWA) is developed to analyze the diffraction properties of acoustic parity-time-symmetric gratings. Asymmetric diffraction can be generated in these gratings, and the +1st order transmitted wave is suppressed completely at the diffraction vanishing point (DVP). The influence factors of the modulation ratio of the DVP, including the modulation amplitude and modulation approach, are systematically discussed using the method of RCWA. Two-beam diffraction with Bragg incident angle is studied in detail. The results show that the modulation ratio of the DVP changes with the modulation amplitude differently for different modulation approaches. The attenuation or enhancement of the sound energy depends on the sign of incident angle and the modulation ratio.

Noggin Inhibits IL-1ß and BMP-2 Expression, and Attenuates Cartilage Degeneration and Subchondral Bone Destruction in Experimental Osteoarthritis.

Osteoarthritis (OA) is a chronic inflammatory and progressive joint disease that results in cartilage degradation and subchondral bone remodeling. The proinflammatory cytokine interleukin 1 beta (IL-1ß) is abundantly expressed in OA and plays a crucial role in cartilage remodeling, although its role in the activity of chondrocytes in cartilage and subchondral remodeling remains unclear. In this study, stimulating chondrogenic ATDC5 cells with IL-1ß increased the levels of bone morphogenetic protein 2 (BMP-2), promoted articular cartilage degradation, and enhanced structural remodeling. Immunohistochemistry staining and microcomputed tomography imaging of the subchondral trabecular bone region in the experimental OA rat model revealed that the OA disease promotes levels of IL-1ß, BMP-2, and matrix metalloproteinase 13 (MMP-13) expression in the articular cartilage and enhances subchondral bone remodeling. The intra-articular injection of Noggin protein (a BMP-2 inhibitor) attenuated subchondral bone remodeling and disease progression in OA rats. We also found that IL-1ß increased BMP-2 expression by activating the mitogen-activated protein kinase (MEK), extracellular signal-regulated kinase (ERK), and specificity protein 1 (Sp1) signaling pathways. We conclude that IL-1ß promotes BMP-2 expression in chondrocytes via the MEK/ERK/Sp1 signaling pathways. The administration of Noggin protein reduces the expression of IL-1ß and BMP-2, which prevents cartilage degeneration and OA development.

Sound Localization and Separation in 3D Space Using a Single Microphone with a Metamaterial Enclosure.

Conventional approaches to sound localization and separation are based on microphone arrays in artificial systems. Inspired by the selective perception of the human auditory system, a multisource listening system which can separate simultaneous overlapping sounds and localize the sound sources in 3D space, using only a single microphone with a metamaterial enclosure is designed. The enclosure modifies the frequency response of the microphone in a direction-dependent manner by giving each direction a characteristic signature. Thus, the information about the location and the audio content of sound sources can be experimentally reconstructed from the modulated mixed signals using a compressive sensing algorithm. Due to the low computational complexity of the proposed reconstruction algorithm, the designed system can also be applied in source identification and tracking. The effectiveness of the system in multiple real-life scenarios is evaluated through multiple random listening tests. The proposed metamaterial-based single-sensor listening system opens a new way of sound localization and separation, which can be applied to intelligent scene monitoring and robot audition.

Design and demonstration of an acoustic right-angle bend.

In this paper, a broadband acoustic right-angle bend device in air is designed, fabricated and experimentally characterized. Perforated panels with various hole-sizes are used to construct the bend structure. Both the simulated and experimental results verify that the acoustic beam can be rotated effectively through the acoustic bend in a wide frequency range. This model may have potential applications in some areas such as sound absorption and acoustic detection in elbow pipes.

Band Offset Measurements in Atomic-Layer-Deposited Al2O3/Zn0.8Al0.2O Heterojunction Studied by X-ray Photoelectron Spectroscopy.

Pure aluminum oxide (Al2O3) and zinc aluminum oxide (Zn x Al1-x O) thin films were deposited by atomic layer deposition (ALD). The microstructure and optical band gaps (E g ) of the Zn x Al1-x O (0.2 ≤ x ≤ 1) films were studied by X-ray diffractometer and Tauc method. The band offsets and alignment of atomic-layer-deposited Al2O3/Zn0.8Al0.2O heterojunction were investigated in detail using charge-corrected X-ray photoelectron spectroscopy. In this work, different methodologies were adopted to recover the actual position of the core levels in insulator materials which were easily affected by differential charging phenomena. Valence band offset (ΔE V) and conduction band offset (ΔE C) for the interface of the Al2O3/Zn0.8Al0.2O heterojunction have been constructed. An accurate value of ΔE V = 0.82 ± 0.12 eV was obtained from various combinations of core levels of heterojunction with varied Al2O3 thickness. Given the experimental E g of 6.8 eV for Al2O3 and 5.29 eV for Zn0.8Al0.2O, a type-I heterojunction with a ΔE C of 0.69 ± 0.12 eV was found. The precise determination of the band alignment of Al2O3/Zn0.8Al0.2O heterojunction is of particular importance for gaining insight to the design of various electronic devices based on such heterointerface.

Oxidative stress-induced 1, N6-ethenodeoxyadenosine adduct formation contributes to hepatocarcinogenesis.

Numerous studies have found that oxidative stress-derived 1, N6-ethenodeoxyadenosine (ε-dA) can act as a driving force towards hepatocellular carcinoma (HCC) in cancer-prone liver diseases. The aim of the present study was to determine the oxidative stress status and the occurrence of ε-dA in HCC and adjacent non-tumor liver tissue, and to clarify whether the occurrence of ε-dA is related to liver inflammatory activity, fibrosis and mutant p53 expression. Oxidative stress-related parameters were examined in tumor and (or) non-tumor liver tissues of 32 patients with HCC. ε-dA, mutant p53 and proliferating cell nuclear antigen (PCNA) were immunohistochemically investigated in control, HCC and non-tumor liver tissues. The total antioxidant capacity and total superoxide dismutase activity of HCC tissues were lower compared to those of non-tumor tissues (P<0.05 vs. P<0.001). The prevalence of ε-dA in HCC was significantly higher compared to control (P<0.0001) and non-tumor liver tissues (P<0.001). A significant correlation between the positive rate of ε-dA and mutant p53 was observed (r=0.5162, P<0.01). The positive rate of PCNA in HCC was significantly higher compared to control (P<0.0001) and non-tumor liver tissues (P<0.0001). There was a possible link between the formation of ε-dA and chronic inflammation and fibrosis. Therefore, ε-dA lesions may gradually accumulate in chronic liver diseases, and partially contribute to mutant p53 overexpression and excessive cell proliferation, making it a potential mechanism in oxidative stress-mediated hepatocarcinogenesis.

Decreased levels of nitric oxide production and nitric oxide synthase-2 expression are associated with the development and metastasis of hepatocellular carcinoma.

Many studies have demonstrated the function of nitric oxide (NO) or nitric oxide synthase-2 (NOS-2) in cancer as pro-neoplastic or anti-neoplastic effectors, but the role of NO and NOS-2 in hepatocellular carcinoma (HCC) remains unclear. The aim of this study was to investigate the levels of NO production and NOS-2 expression in HCC and adjacent non-tumor liver tissues and to clarify whether the levels of NO/NOS-2 are related to the clinicopathological features of HCC. The levels of NO production were examined in tumor and adjacent non-tumor liver tissues of 30 patients with HCC. The expression of NOS-2 was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemical analysis in HCC and/or adjacent non-tumor liver tissues. Mutant p53 and proliferating cell nuclear antigen (PCNA) were also immunohistochemically investigated in liver tissues. The levels of NO in HCC were significantly lower compared to adjacent non-tumor liver tissues (P<0.001). The relative mRNA and protein expression levels of NOS-2 in HCC were also significantly lower compared to adjacent non-tumor liver tissues (P<0.01 for both). We found that the levels of NO in patients suffering from HCC metastasis were lower compared to those without metastasis (P<0.05) and NOS-2 expression was correlated with tumor diameter (P<0.05) and metastasis (P<0.05). In addition, mutant p53 protein was expressed in the majority of HCC samples and the proliferation rate of HCC was significantly higher than that of adjacent non-tumor liver tissues. These data indicate that decreased levels of NO/NOS-2 may partially contribute to overexpression of the mutant p53 protein and excessive proliferation; this may be a potential mechanism in the development and progression of HCC.

Hepatitis B virus X protein promotes the growth of hepatocellular carcinoma by modulation of the Notch signaling pathway.

Hepatitis B virus X protein (HBx) plays a crucial role in the development of hepatocellular carcinoma (HCC), however, little is known about the mechanism. Here, we investigated the relationship between HBx and Notch signaling in HepG2 cells after they were transfected with the HBx gene. It was found that HBx upregulated the expression of Notch-1, Jagged-1 and Hes-1 at the transcriptional level by binding to the Notch-1 intracellular domain, which is congruent with the observations of enhanced malignant biological activities of HBx-transfected HepG2 cells compared with normal HepG2 cells. However, while Notch signaling was blocked, the HBx-induced abnormalities were partially reversed. These findings suggest that HBx may promote the progression of HCC via the activated Notch pathway.

[Analysis on the septum correction and plasma radiofrequency ablation combination therapy for nasal septum deviation].

OBJECTIVE: To discuss a effective method of nasal septum deviation. METHOD: One hundred and sixteen subjects with nasal septum deviation were divided into treatment group (69 subjects) and control group (47 subjects) randomly. The combination therapy of correction of deviated nasal septum and plasma radiofrequency ablation were used in the treatment group. The combination therapy of deviated nasal septum and partial inferior turbinectomy were used in the control group. The data were analyzed by statistical method. RESULT: The effective rate of physical signs and symptoms of the treatment group was 100.0%, while the control group was 85.1%. There was significant difference of two effective rates (P < 0.05). Furthermore, hyperventilation and nasal adhesion were not happened in treatment group 6 months after treatments. CONCLUSION: The correction of deviated nasal septum and plasma radiofrequency ablation combination therapy had the satisfied and safety treatment effect, which was and plasma radiofrequency ablation combination therapy had the satisfied and safety treatment effect, which was easy for observation as well as the minimal tissue damage. The combination therapy method was according with the principle of functional minimally invasive surgery.