Genome-Wide Association and Expression Analysis of the Lipoxygenase Gene Family in Passiflora edulis Revealing PeLOX4 Might Be Involved in Fruit Ripeness and Ester Formation.

Aroma is an important factor in fruit quality. Passiflora edulis (passion fruit) is popular among consumers because of its rich flavor and nutritional value. Esters are the main components of the volatile aroma of passion fruit. Lipoxygenase (LOX), as the first key enzyme upstream of esters, may play an important role in the formation of passion fruit aroma. In this study, a total of 12 passion fruit LOX (PeLOX) members were screened out based on the Passiflora edulis genome database, which were distributed unevenly on 6 chromosomes, all containing the highly conserved lipoxygenase domain and some containing the PLAT domain. The gene structure, evolutionary analysis and cis-acting elements of the family members were predicted in this study. Transcriptome analysis showed that 12 PeLOX genes had different degrees of response to different abiotic stresses (drought stress, salt stress, cold stress, and high temperature). PeLOX1, PeLOX2, PeLOX7, PeLOX11, and PeLOX12 responded significantly to various abiotic stresses, while PeLOX8 and PeLOX9 had little change in expression in all stresses. Quantitative real-time PCR (qRT-PCR) in six tissues revealed that the 12 PeLOX genes exhibited tissue expression specificity, and the relative expression of most genes were particularly high in the roots, stems, and fruits. Focusing on passion fruit ripening and ester synthesis, the transcriptomic analysis showed that with the increase in fruit development and fruit maturity, the expression levels of PeLOX1, PeLOX9, PeLOX11, and PeLOX12 showed downregulated expression, while PeLOX2 and PeLOX4 showed upregulated expression. In particular, the upregulation trend of PeLOX4 was the most obvious, and the qRT-PCR results were consistent with the transcriptome result. Pearson correlation analysis showed that with the development and ripening of fruit, the expression level of PeLOX4, LOX enzyme activity and total ester content all showed an increasing trend, in particular during the period when the peel was red and shrank (from T2 to T3 stage), the esters' contents increased by 37.4 times; the highest expression levels were all in the T3 period. The results indicated that PeLOX4 may be a candidate gene involved in fruit ripeness and the formation of volatile aroma compounds, with the increase in fruit ripening, the expression level of PeLOX4 increased and the LOX enzyme activity increased accordingly, thereby promoting the synthesis of volatile esters in fruit pulp. Our discovery lays the foundation for the functional study of LOX in passion fruit.

HSP70 alleviates spinal cord injury by activating the NF-kB pathway.

OBJECTIVES: Spinal cord injury (SCI) is an acute traumatic lesion of neurons in the spinal cord which has a high prevalence in the world, and has no effective surgical treatment. HSP70 is a molecular chaperone protein, serves a protective role in several different models of nervous system injury. The aim of the present study was to investigate the anti-inflammatory role of HSP70 in spinal cord injury and explore its mechanism. METHODS: In vivo and in vitro models were constructed to mimic SCI. The Basso Mouse Scale (BMS) was applied to assess SCI degrees of the mouse model. Immunofluorescence (IF) was used for visualizing HSP70 and Iba1 in the spinal cord. Western blot assay was employed to quantify HSP70 and p65, and ELISA was for IL-1ß and TNF-α. RESULTS: The results showed that HSP70 expression decreased after SCI. HSP70 and Iba1 showed a decrease of co-localization in SCI mice. Further studies revealed that p65 was upregulated during the process of SCI. Overexpression of HSP70 inhibited the expression of p65 both in vitro and in vivo, and promoted the recovery of SCI mice. CONCLUSIONS: HSP70 was involved in the pathological process of spinal cord injury, HSP70 alleviated the spinal cord injury via inhibiting NF-κB signaling pathway.

S100-ß aggravates spinal cord injury via activation of M1 macrophage phenotype.

OBJECTIVES: S100-ß has been identified as a sensitive biomarker in central nervous system injuries. However, the functions and mechanisms of S100-ß are unknown in spinal cord injury. METHODS: Spinal cord injury (SCI) mouse model was generated by surgical operation, microglia activation model was established by inducing BV-2 cells with LPS. The SCI model was evaluated by Basso-Beattie-Bresnahan (BBB) behavioral score, HE staining, and Nissl staining. The expression level of S100-ß was detected by qRT-PCR, western blot, and immunofluorescence. qRT-PCR and western blot were used to detect the expression of iNOS and CD16. Pro-inflammatory cytokines TNF-α and IL-1ß levels were detected by qRT-PCR and ELISA. RESULTS: The expression of IL-1ß, TNF-α, iNOS, and CD16 increased at 3rd day after SCI. In BV2 microglia, LPS treatment promoted the expression of S100-ß, IL-1ß, TNF-α, iNOS, and CD16. Knockdown of S100-ß reduced the expression of iNOS stimulated by LPS. Over-expression of S100-ß increased IL-1ß and TNF-α, and S100-ß inhibition suppressed IL-1ß and TNF-α. In SCI mice, knockdown of S100-ß attenuated the spinal cord injury and inhibited the expression of iNOS, IL-1ß, and TNF-α. CONCLUSIONS: Down-regulation of S100-ß could inhibit the pathogenesis of SCI and inhibit the activation of M1 macrophages. S100-ß may be a useful diagnostic biomarker or therapeutic target for SCI.

Genome-wide analysis and phosphorylation sites identification of the 14-3-3 gene family and functional characterization of MeGRF3 in cassava.

The biological functionality of many members of the 14-3-3 gene family is regulated via phosphorylation at multiple amino acid residues. The specific phosphorylation-mediated regulation of these proteins during cassava root tuberization, however, is not well understood. In this study, 15 different 14-3-3 genes (designated MeGRF1 - 15) were identified within the cassava genome. Based upon evolutionary conservation and structural analyses, these cassava 14-3-3 proteins were grouped into ε and non-ε clusters. We found these 15 MeGRF genes to be unevenly distributed across the eight cassava chromosomes. When comparing the expression of these genes during different developmental stages, we found that three of these genes (MeGRF9, 12 and 15) were overexpressed at all developmental stages at 75, 104, 135, 182 and 267 days post-planting relative to the fibrous root stage, whereas two (MeGRF5 and 7) were downregulated during these same points. In addition, the expression of most MeGRF genes changed significantly in the early and middle stages of root tuberization. This suggests that these different MeGRF genes likely play distinct regulatory roles during cassava root tuberization. Subsequently, 18 phosphorylated amino acid residues were detected on nine of these MeGRF proteins. A phosphomimetic mutation at serine-65 in MeGRF3 in Arabidopsis thaliana (Arabidopsis) slightly influenced starch metabolism in these plants, and significantly affected the role of MeGRF3 in salt stress responses. Together these results indicate that 14-3-3 genes play key roles in responses to abiotic stress and the regulation of starch metabolism, offering valuable insights into the functions of these genes in cassava.

The Three-Dimensional Computed Tomography Anatomic Features of Internal Carotid Artery-Ophthalmic Artery.

OBJECTIVE: This research aimed at exploring the measurement of anatomic parameters of internal carotid artery-ophthalmic artery (ICA-OA) and the clinical significance with 3-dimensional (3D) separating, fusing, opacifying, false-coloring, and volume rendering (SFOF-VR) technique. METHODS: A total of 109 patients with no significant lesions in ICA-OA were randomly selected, including 56 males and 53 females, who were divided into 3 groups: young age group (18-40 years, n = 36), middle age group (41-60 years, n = 40) and old age group (>60 years, n = 33). The 3D images were constructed by AW 4.6 workstation, and the anatomic parameters were measured. The SFOF-VR technique explicitly displayed ICA, OA, and their concomitant relationship with ophthalmic nerves (ONs) and adjacent structures. The study protocol was approved by the Medical Ethical Committee of Shaanxi Provincial People's Hospital and all patients have signed consent forms. RESULTS: The diameters of initial segment of left and right ophthalmic ICA (ICA-C6) were 3.58 ±â€Š0.51 mm and 3.58 ±â€Š0.43 mm, respectively; the diameters of the left and right OA were 1.16 ±â€Š0.16 mm and 1.15 ±â€Š0.19 mm, respectively; the lengths of left and right ICA-C6 were 10.43 ±â€Š2.08 mm and 10.01 ±â€Š2.32 mm, respectively. There were statistically significant differences between bilateral ICA-C6 lengths, right ICA-C6 diameter, right OA diameter, and OA-ON space between male and female. CONCLUSION: The SFOF-VR technique can distinctly present the structures of ICA-OA and adjacent tissues, providing valuable information for surgical operation. The 3D computed tomography data about ICA-OA can enrich references for basic and clinical researches.

Multimodal imaging in the diagnosis of bone giant cell tumors: A retrospective study.

BACKGROUND: Giant cell tumor of bone is a locally aggressive and rarely metastasizing tumor, and also a potential malignant tumor that may develop into a primary malignant giant cell tumor. AIM: To evaluate the role of multimodal imaging in the diagnosis of giant cell tumors of bone. METHODS: The data of 32 patients with giant cell tumor of bone confirmed by core-needle biopsy or surgical pathology at our hospital between March 2018 and March 2023 were retrospectively selected. All the patients with giant cell tumors of the bone were examined by X-ray, computed tomography (CT) and magnetic resonance imaging (MRI), and 7 of them were examined by positron emission tomography (PET)-CT. RESULTS: X-ray imaging can provide overall information on giant cell tumor lesions. CT and MRI can reveal the characteristics of the internal structure of the tumor as well as the adjacent relationships of the tumor, and these methods have unique advantages for diagnosing tumors and determining the scope of surgery. PET-CT can detect small lesions and is highly valuable for identifying benign and malignant tumors to aid in the early diagnosis of metastasis. CONCLUSION: Multimodal imaging plays an important role in the diagnosis of giant cell tumor of bone and can provide a reference for the treatment of giant cell tumors.

Diagnostic value of artificial intelligence based on computed tomography (CT) density in benign and malignant pulmonary nodules: a retrospective investigation.

Objective: To evaluate the diagnostic value of artificial intelligence (AI) in the detection and management of benign and malignant pulmonary nodules (PNs) using computed tomography (CT) density. Methods: A retrospective analysis was conducted on the clinical data of 130 individuals diagnosed with PNs based on pathological confirmation. The utilization of AI and physicians has been employed in the diagnostic process of distinguishing benign and malignant PNs. The CT images depicting PNs were integrated into AI-based software. The gold standard for evaluating the accuracy of AI diagnosis software and physician interpretation was the pathological diagnosis. Results: Out of 226 PNs screened from 130 patients diagnosed by AI and physician reading based on CT, 147 were confirmed by pathology. AI had a sensitivity of 94.69% and radiologists had a sensitivity of 85.40% in identifying PNs. The chi-square analysis indicated that the screening capacity of AI was superior to that of physician reading, with statistical significance (p < 0.05). 195 of the 214 PNs suggested by AI were confirmed pathologically as malignant, and 19 were identified as benign; among the 29 PNs suggested by AI as low risk, 13 were confirmed pathologically as malignant, and 16 were identified as benign. From the physician reading, 193 PNs were identified as malignant, 183 were confirmed malignant by pathology, and 10 appeared benign. Physician reading also identified 30 low-risk PNs, 19 of which were pathologically malignant and 11 benign. The physician readings and AI had kappa values of 0.432 and 0.547, respectively. The physician reading and AI area under curves (AUCs) were 0.814 and 0.798, respectively. Both of the diagnostic techniques had worthy diagnostic value, as indicated by their AUCs of >0.7. Conclusion: It is anticipated that the use of AI-based CT diagnosis in the detection of PNs would increase the precision in early detection of lung carcinoma, as well as yield more precise evidence for clinical management.

A method for high-concentration agarose gel preparation and its application in high-resolution separation of low-molecular-weight nucleic acids and proteins.

Separation of nucleic acids and proteins using gels has always been a crucial part of molecular biology research. For low-molecular-weight nucleic acids and proteins, low- and medium-concentration agarose gels cannot achieve the high resolution as polyacrylamide gels. We found that 6 %-14 % high-concentration agarose gels (HAGs) could be easily dissolved in an autoclave and the vertical gel cast can be effortlessly filled using an easy-made plastic box. Coupled with the improved buffer condition, HAG electrophoresis resulted in a good resolution of DNA and protein bands. With conventional TBE buffer plus 0.2 % NaCl, DNA fragments that differ by 2-5-bp within the 50-200-bp size range can be resolved on 6 %-8 % HAGs. By using TBE without NaCl, DNA fragments that differ by 2-bp or 2-nt within the 10-100-bp size range can be well resolved on >8 % HAGs. Using a buffer system comprising 1 M Tris-Cl for gel preparation, 0.2 M Tris-Cl/0.2 % SDS as upper tank buffer, and 0.2 M Tris-Cl as the lower tank buffer, HAGs achieved good molecular weight separation of total bacterial and plant proteins in the 10-200 kDa range. In conclusion, we developed a method for HAG preparation and electrophoresis of low-molecular-weight nucleic acids and proteins.

Functional disparity of four pheromone-binding proteins from the plum fruit moth Grapholita funebrana Treitscheke in detection of sex pheromone components.

Grapholita funebrana, also known as the plum fruit moth, is an oligophagous pest species that causes enormous economic losses of the fruits of Rosaceae. An eco-friendly method for the control of G. funebrana besides chemical control has not yet been developed. The sex pheromone communication system plays an important role in moth courtship and mating, in which pheromone-binding proteins (PBPs) are critical. In this research, we identified four PBPs, namely, GfunPBP1.1, GfunPBP1.2, GfunPBP2, and GfunPBP3, from the antennae of G. funebrana. The results of real-time quantitative PCR (RT-qPCR) showed that all four GfunPBPs were overwhelmingly expressed in the antennae and that GfunPBP1.2 and GfunPBP2 showed male-biased expression patterns, whereas GfunPBP1.1 and GfunPBP3 were equally expressed between sexes. The results of ligand-binding assays illustrated that although all four recombinant GfunPBPs (rGfunPBPs) had binding activity with the tested sex pheromone compounds, their preferred ligands were significantly different. rGfunPBP2 had the strongest binding affinity to Z8-12:Ac and Z8-12:OH; rGfunPBP1.1 preferred to bind Z8-14:Ac, Z10-14:Ac, and 12:OH more than to the other three GfunPBPs; and rGfunPBP1.2 exhibited stronger binding affinity to E8-12:Ac than to the other rGfunPBPs. Molecular docking results demonstrated that hydrophobic forces, especially van der Waals forces and hydrogen bonds, were the most important forces that maintained GfunPBP-pheromone ligand complexes. This study will improve our understanding of the sex pheromone recognition mechanisms of G. funebrana and promote the development of novel strategies for controlling G. funebrana.

Comprehensive analysis of bZIP transcription factors in passion fruit.

The bZIP transcription factors are well-known transcriptional regulators that are essential for regulating resistance to biotic and abiotic stresses in plants. In this study, a total of 56 putative bZIP members were identified in passion fruit (Passiflora edulis). An integrative analysis was performed using bioinformatics. Transcriptome analysis revealed that most PebZIPs respond to drought, salt, cold and heat stress. By combining the transcriptome results of two different resistant genotypes, four representative members were finally selected for differential expression validation in different tissues and cultivars. Furthermore, transcriptome and metabolome association analysis revealed consistent expression trends of PeZIP20 and PeZIP21, with only one difference at 63aa, with different metabolites including flavonoids, lipids and amino acids. This work will contribute to further studies of the functions of bZIPs and their resistance properties, as well as to the development of novel germplasm.

Application of CT Postprocessing Reconstruction Technique in Differential Diagnosis of Benign and Malignant Solitary Pulmonary Nodules and Analysis of Risk Factors.

Objective: To evaluate the application of CT postprocessing reconstruction technique in differential diagnosis of benign and malignant solitary pulmonary nodules and analysis of risk factors. Methods: A total of 150 solitary pulmonary nodules (SPN) patients admitted to our hospital from January 2020 to January 2022 were selected and divided into the benign SPN group (n = 64) and the malignant SPN group (n = 86) according to pathological results. All subjects underwent CT plain scan and CT postprocessing reconstruction, and the general information of the subjects was collected. The diagnostic value of CT plain scan and CT postprocessing reconstruction techniques for benign and malignant SPN was compared; and the CT signs of benign and malignant SPN were compared, and the risk factors of malignant SPN were analyzed. Results: The pathological results of this study showed that there were 64 cases with benign SPN and 86 cases with malignant SPN. The sensitivity, specificity, accuracy, positive predictive rate, and negative predictive rate of CT postprocessing reconstruction technology in diagnosing malignant SPN were 73.44%, 89.53%, 82.67%, 83.39%, and 81.91%, respectively, which were higher than 56.25%, 65.12%, 61.33%, 54.55%, and 66.67% of CT plain scan, and the difference was statistically significant (P < 0.05). There were no significant differences in nodule location, nodule density, vacuole sign, vessel convergence, and pleural depression sign between the two groups (P > 0.05). There were statistically significant differences in age, nodule diameter, lobulation sign, burr sign, calcification components, and ground-glass components between the two groups (P < 0.05). Multivariate analysis showed that age ≥ 60 years, nodule diameter ≥ 15 mm, the presence of lobulation sign, burr sign, ground-glass components, and noncalcification components were independent risk factors for malignant SPN. Conclusion: CT postprocessing reconstruction technique has high diagnostic value in the differentiation of benign and malignant SPN, age ≥ 60 years, nodule diameter ≥ 15 mm, lobulation signs, burr signs, ground-glass components, and noncalcification components are independent risk factors for malignant SPN.

Puerarin attenuates cisplatin-induced apoptosis of hair cells through the mitochondrial apoptotic pathway.

Puerarin, one of the main components of Pueraria lobata, has been reported to possess a wide range of pharmacological activities, including anti-inflammatory, antioxidative and anti-apoptotic effects. However, the role of puerarin in ototoxic drug-induced hair cell injury has not been well characterized. This study explored whether puerarin protects against cisplatin-induced hair cell damage and its potential mechanisms. The viability of puerarin-treated HEI-OC1 cells was assessed by CCK8 assay. Reactive oxygen species (ROS) was estimated with flow cytometric analysis using Cellrox Green fluorescent probe. Apoptosis-related protein levels were detected by western blot analysis. Immunostaining of the organ of Corti was performed to determine mice cochlear hair cell survival. Our results showed that puerarin improved cell viability and suppressed apoptosis in the cisplatin-damaged HEI-OC1 cells and cochlear hair cells. Mechanistic studies revealed that puerarin attenuated mitochondrial apoptosis pathway by regulating apoptotic related proteins, such as Bax and cleaved caspase-3, and attenuated ROS accumulation after cisplatin damage. Moreover, puerarin was involved in regulating the Akt pathway in HEI-OC1 cells in response to cisplatin. Our results demonstrated that puerarin administration decreased the sensitivity to apoptosis dependent on the mitochondrial apoptotic pathway by reducing ROS generation, which could be used as a new protective agent against cisplatin-induced ototoxicity.

Efficacy and Safety Analysis of Multislice Spiral CT-Guided Transthoracic Lung Biopsy in the Diagnosis of Pulmonary Nodules of Different Sizes.

Objective: This study is aimed at investigating the efficacy and safety of multislice spiral CT-guided transthoracic lung biopsy in the diagnosis of pulmonary nodules of different sizes. Methods: Data of 78 patients with pulmonary nodules who underwent CT-guided transthoracic lung biopsy in our hospital from January 2020 to December 2021 were retrospectively analyzed, and they were divided into the small nodules group (n = 12), medium nodules group (n = 35), and large nodules group (n = 31) according to the diameter of pulmonary nodules. The results of puncture biopsy and final diagnosis of pulmonary nodules of different sizes were compared. The incidence of complications in patients with pulmonary nodules of different sizes was compared. Univariate analysis was used to compare the incidence of complications in 78 patients. Logistic multiple regression analysis was used to analyze the independent risk factors of pneumothorax in patients with pulmonary nodule puncture. Logistic multiple regression analysis was used to analyze the independent risk factors of pulmonary hemorrhage in patients with pulmonary nodule puncture. Results: The diagnostic accuracy, sensitivity, and specificity were 83.33%, 100.00%, and 77.78% in small nodules group. The diagnostic accuracy, sensitivity, and specificity of medium nodules group were 85.71%, 100.00%, and 73.68%, respectively. The diagnostic accuracy, sensitivity, and specificity of large nodules group were 93.55%, 100.00%, and 33.33%, respectively. There was no significant difference in the incidence of pneumothorax among the three groups (P > 0.05). The incidence of pulmonary hemorrhage in small nodule group was higher than that in the medium nodule group and large nodule group, and the difference was statistically significant (P < 0.05). There was no significant difference in the incidence of total complications among the three groups (P > 0.05). There were statistically significant differences in clinical data such as the needle tract length, the puncture position, and the distance of the puncture needle passing through the lung tissue in patients with or without pneumothorax (P < 0.05). There were statistically significant differences in needle tract length, distance of puncture needle passing through lung tissue, and size of pulmonary nodules in patients with or without pulmonary hemorrhage (P > 0.05). Logistic multivariate analysis showed that needle tract length ≤ 50 mm, lateral decubitus position, and the distance of puncture needle passing through lung tissue ≥ 14 mm were independent risk factors for pneumothorax after puncture in patients with pulmonary nodules (P < 0.05). The needle tract length > 50 mm, the distance of puncture needle passing through lung tissue ≥ 14 mm, and small nodules (pulmonary nodules diameter ≤ 10 mm) were independent risk factors for pulmonary hemorrhage after puncture in patients with pulmonary nodules (P < 0.05). Conclusion: Multislice spiral CT-guided transthoracic lung biopsy is effective in diagnosing pulmonary nodules of different sizes.

Different Binding Affinities of Three General Odorant-Binding Proteins in Grapholita funebrana (Treitscheke) (Lepidoptera: Tortricidae) to Sex Pheromones, Host Plant Volatiles, and Insecticides.

Insect general odorant-binding proteins (GOBPs) play irreplaceable roles in filtering, binding, and transporting host odorants to olfactory receptors. Grapholita funebrana (Treitscheke) (Lepidoptera: Tortricidae), an economically important pest of fruit crops, uses fruit volatiles as cues to locate host plants. However, the functions of GOBPs in G. funebrana are still unknown. Three GOBP genes, namely, GfunGOBP1, GfunGOBP2, and GfunGOBP3, were cloned, and their expression profiles in different tissues were detected by the method of real-time quantitative PCR (RT-qPCR). The binding properties of recombinant GfunGOBPs (rGfunGOBPs) to various ligands were investigated via fluorescence binding assays. The three GfunGOBPs were mainly expressed in the antennae of both male and female moths. All these three rGfunGOBPs could bind to sex pheromones, while having varying affinities toward these pheromones. The three rGfunGOBPs also displayed a wide range of ligand-binding spectrums with tested host odorants. The rGfunGOBP1, rGfunGOBP2, and rGfunGOBP3 bound to 34, 33, and 30 out of the 41 tested odorants, respectively. Three rGfunGOBPs had overlapping binding activities to ß-myrcene, (-)-α-phellandrene, and ethyl isovalerate with the Ki less than 3.0 µM. The rGfunGOBP1 and rGfunGOBP3 could selectively bind to several insecticides, whereas rGfunGOBP2 could not. Three rGfunGOBPs had the dual functions of selectively binding to sex pheromones and host odorants. Moreover, the rGfunGOBP1 and rGfunGOBP3 can also serve as 'signal proteins' and bind to different insecticides. This study contributed to elucidating the potential molecular mechanism of the olfaction for G. funebrana, and thereby promotes the development of effective botanical attractants or pheromone synergists to control G. funebrana.

The complete chloroplast genome of Arcangelisia gusanlung H.S.Lo (Menispermaceae), an important traditional medicine from South China.

Arcangelisia gusanlung H.S.Lo is widely used as a folk medicine by the Dai and Li peoples. Here, we report the first complete chloroplast (cp) genome sequence for this species based on Illumina paired-end sequencing data. The cp genome was 162,509 bp in length with a small single-copy (SSC) region of 20,852 bp, a large single-copy (LSC) region of 91,449 bp, and two separated inverted region of 25,104 bp. In total, 129 unique genes were identified of this genome, including 84 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The GC contents of this genome is 37.8%. Phylogenetic analysis based on 13 complete cp genomes showed a strong sister relationship with Tinospora cordifolia (Willd.) Miers and Tinospora sinensis (Lour.) Merr. This complete genome of A. gusanlung will provide valuable information to elucidate the mechanism of speciation of Arcangelisia Becc.

An Imaging Measurement Study of Normal Knee Parameters in Southeast China.

BACKGROUND: Imaging measurement of distal femur and proximal tibia has been the hot point in the research of total knee arthroplasty and prosthesis development, which is an important treatment for patients with advanced knee joint disease. This study retrospectively investigated the digital imaging measurement of normal knee parameters in southeast China and evaluated their clinical value. METHODS: From February 2010 to May 2014, and in accordance with the inclusion criteria, a total of 677 knees (334 female knees and 343 male knees) were categorized into 3 age groups. Clinical and digital imaging data, including the distal Femoral Condyle Diameter (FCD), Tibial Plateau Diameter (TPD), the distance between the medial tibial plateau and fibular head (DPF), tibiofemoral valgus angle, distal femoral valgus angle, Proximal Tibia (PT) varus angle and the angle from femoral condyle to tibial perpendicular (FT), were measured by using AutoCAD 10.0 software. All measured variables were statistically analyzed by SPSS statistical software (version 18.0). RESULTS: Data are presented as the mean ± standard deviation. The normal female and male femoral condyle diameter was (7.69 ± 0.46) cm and (8.68 ± 0.55) cm, while the normal female and male tibial plateau diameter was (7.66 ± 0.46) cm and (8.60 ± 0.55) cm, respectively. The normal female and male DPF was (0.76 ± 0.36) cm and (0.79 ± 0.36) cm. For females and males, the tibiofemoral valgus angle and distal femoral valgus angle were (3.89 ± 2.20) ° and (3.29 ± 2.12) °, (9.03 ± 2.18) ° and (8.25 ± 2.20) °. As the two methods to measure tibial plateau varus angle, PT angle of normal female and male was (4.29 ± 1.86) ° and (4.84 ± 2.23) °, while the normal female and male FT angle was (5.34 ± 1.95) ° and (5.52 ± 2.07) °. Based on the data obtained, we found significant differences between the two genders in terms of the femoral condyle diameter and tibial plateau diameter in all age groups (P < 0.01). The DPF parameter showed an obvious difference between the young group and the middle-aged group (P < 0.05), and no significant difference was observed between the sides and genders (P > 0.05). The distal femoral valgus angle showed statistical differences between genders in the left side of the young group and middle-aged group (P < 0.05), while angle PT and FT showed no significant difference (P > 0.05). CONCLUSION: A large number of knee measurements was obtained, and a local knee database was developed in this study. Imaging measurement prior to total knee arthroplasty is clinically important for increasing the accuracy and long-term efficacy of total knee arthroplasty. These data can also provide useful information for knee surgery and sports medicine as well as prosthesis development.

Circ_0081001 down-regulates miR-494-3p to enhance BACH1 expression and promotes osteosarcoma progression.

The study was aimed at deciphering the function and mechanism of circ_0081001 in osteosarcoma (OS). In this study, quantitative real-time polymerase chain reaction (qRT-PCR) was utilized for quantifying circ_0081001, miR-494-3p, and BTB domain and CNC homolog 1 (BACH1) mRNA expressions in OS tissues and cells. Cell counting kit-8 (CCK-8) assay, together with 5-Ethynyl-2'-deoxyuridine (EdU) assay, was performed for evaluating cell proliferation; the alterations in apoptosis were analyzed utilizing flow cytometry; Transwell assay was conducted for examining cell migration and invasion; moreover, Western blot was utilized for the quantification of BACH1 protein expression; bioinformatics, dual-luciferase reporter gene, and RNA-binding protein immunoprecipitation assays were executed for validating the binding relationships between circ_0081001 and miR-494-3p, and between miR-494-3p and BACH1. As shown, circ_0081001, whose expression was elevated in OS tissues, had a negative association with miR-494-3p expression and a positive correlation with BACH1 expression. After circ_0081001 was overexpressed, the proliferation, migration, and invasion of OS cells were boosted but the apoptosis was reduced, whereas miR-494-3p exhibited opposite effects. The binding sites between circ_0081001 and miR-494-3p, and between miR-494-3p and the 3'UTR of BACH1 were experimentally verified. In conclusion, circ_0081001/miR-494-3p/BACH1 axis promoted the malignant biological behaviors of OS cells.

Identification of Candidate Olfactory Genes in Scolytus schevyrewi Based on Transcriptomic Analysis.

The bark beetle, Scolytus schevyrewi (S. schevyrewi), is an economically important pest in China that causes serious damage to the fruit industry, particularly, in Xinjiang Province. Chemical signals play an important role in the behavior of most insects, accordingly, ecofriendly traps can be used to monitor and control the target pests in agriculture. In order to lay a foundation for future research on chemical communication mechanisms at the molecular level, we generate antennal transcriptome databases for male and female S. schevyrewi using RNA sequencing (RNA-seq) analysis. By assembling and analyzing the adult male and female antennal transcriptomes, we identified 47 odorant receptors (ORs), 22 ionotropic receptors (IRs), 22 odorant-binding proteins (OBPs), and 11 chemosensory proteins (CSPs). Furthermore, expression levels of all the candidate OBPs and CSPs were validated in different tissues of male and female adults by semiquantitative reverse transcription PCR (RT-PCR). ScosOBP2 and ScosOBP18 were highly expressed in female antennae. ScosCSP2, ScosCSP3, and ScosCSP5 were specifically expressed in the antennae of both males and females. These results provide new potential molecular targets to inform and improve future management strategies of S. schevyrewi.

Hsa_circ_0008259 modulates miR-21-5p and PDCD4 expression to restrain osteosarcoma progression.

BACKGROUND: Osteosarcoma (OS) is one of the most common primary bone tumors in children and adolescents. However, the molecular mechanism of OS tumorigenesis is still little known. Circular RNA (circRNA) is a key player in the progression of many cancers. This study is performed to decipher the role and mechanism of circ_0008259 in the progression of OS. METHODS: A differentially expressed circRNA, circ_0008259, was screened out by analyzing the expression profile of circRNA in OS tissue. Circ_0008259, miR-21-5p and programmable cell death 4 (PDCD4) mRNA expression levels in OS tissues and cells were detected by qRT-PCR. Cell viability, metastatic potential and apoptosis were evaluated by cell counting kit-8 assay, Transwell and flow cytometry. The targeting relationship between circ_0008259 and miR-21-5p, and miR-21-5p and PDCD4 mRNA was analyzed and probed by bioinformatics analysis and dual-luciferase reporter assay, RNA-binding protein immunoprecipitation assay and RNA-pull down assay. The regulatory effects of circ_0008259 and miR-21-5p on PDCD4 protein expression in OS cells were detected by Western blot assay. RESULTS: Circ_0008259 expression and PDCD4 expression were down-regulated and miR-21-5p expression was elevated in the OS tissues and cells. Functional experiments showed that circ_0008259 overexpression significantly inhibited the proliferation and metastatic potential of OS cells and promoted the apoptosis. Besides, PDCD4 was validated as the target gene of miR-21-5p, and circ_0008259 could competitively bind to miR-21-5p, thus up-regulating PDCD4 expression in OS cells. CONCLUSIONS: Circ_0008259 suppresses OS progression via regulating miR-21-5p/PDCD4 axis.

Chromosome-scale genome assembly provides insights into the evolution and flavor synthesis of passion fruit (Passiflora edulis Sims).

Passion fruit (Passiflora edulis Sims) is an economically valuable fruit that is cultivated in tropical and subtropical regions of the world. Here, we report an ~1341.7 Mb chromosome-scale genome assembly of passion fruit, with 98.91% (~1327.18 Mb) of the assembly assigned to nine pseudochromosomes. The genome includes 23,171 protein-coding genes, and most of the assembled sequences are repetitive sequences, with long-terminal repeats (LTRs) being the most abundant. Phylogenetic analysis revealed that passion fruit diverged after Brassicaceae and before Euphorbiaceae. Ks analysis showed that two whole-genome duplication events occurred in passion fruit at 65 MYA and 12 MYA, which may have contributed to its large genome size. An integrated analysis of genomic, transcriptomic, and metabolomic data showed that 'alpha-linolenic acid metabolism', 'metabolic pathways', and 'secondary metabolic pathways' were the main pathways involved in the synthesis of important volatile organic compounds (VOCs) in passion fruit, and this analysis identified some candidate genes, including GDP-fucose Transporter 1-like, Tetratricopeptide repeat protein 33, protein NETWORKED 4B isoform X1, and Golgin Subfamily A member 6-like protein 22. In addition, we identified 13 important gene families in fatty acid pathways and eight important gene families in terpene pathways. Gene family analysis showed that the ACX, ADH, ALDH, and HPL gene families, especially ACX13/14/15/20, ADH13/26/33, ALDH1/4/21, and HPL4/6, were the key genes for ester synthesis, while the TPS gene family, especially PeTPS2/3/4/24, was the key gene family for terpene synthesis. This work provides insights into genome evolution and flavor trait biology and offers valuable resources for the improved cultivation of passion fruit.