Development of SSR molecular markers and genetic diversity analysis of Clematis acerifolia from Taihang Mountains.

Investigating the genetic diversity and population structure is important in conserving narrowly distributed plants. In this study, 90 Clematis acerifolia (C. acerifolia) plants belonging to nine populations were collected from the Taihang Mountains in Beijing, Hebei, and Henan. Twenty-nine simple sequence repeats (SSR) markers developed based on RAD-seq data were used to analyze the genetic diversity and population structure of C. acerifolia. The mean PIC value for all markers was 0.2910, indicating all SSR markers showed a moderate degree of polymorphism. The expected heterozygosity of the whole populations was 0.3483, indicating the genetic diversity of both C. acerifolia var. elobata and C. acerifolia were low. The expected heterozygosity of C. acerifolia var. elobata (He = 0.2800) was higher than that of C. acerifolia (He = 0.2614). Genetic structure analysis and principal coordinate analysis demonstrated that C. acerifolia and C. acerifolia var. elobata showed great genetic differences. Molecular variance analysis (AMOVA) demonstrated that within-population genetic variation (68.31%) was the main contributor to the variation of the C. acerifolia populations. Conclusively, C. acerifolia var. elobata had higher genetic diversity than C. acerifolia, and there are significant genetic differences between C. acerifolia and C. acerifolia var. elobata, and small genetic variations within the C. acerifolia populations. Our results provide a scientific and rational basis for the conservation of C. acerifolia and provide a reference for the conservation of other cliff plants.

SVF-GEL Cryopreserved for Different Times Exhibits Varied Preservation and Regeneration Potential After Transplantation in a Mouse Model.

BACKGROUND: Matrix vascular component (SVF) gels derived from fat preserve tissue integrity and cell viability under cryopreserved conditions, making them easy to inject again for later use. Here, we compared the preservation power and regeneration potential of SVF-gel under different cryopreservation times. METHODS: The SVF-gel stored under - 20 °C, without cryoprotectant cryopreservation for 5, 15, and 45 days, with fresh SVF-gel as control. We evaluated the rate of volume retention after thawing the SVF-gel and the apoptosis rate of adipose-derived stem cells. Next, we analyzed retention rated, adipogenesis, angiogenesis, and connective tissue hyperplasia of the grafts, one month after subcutaneously transplanting the specimen into immunodeficient mice. RESULTS: SVF-gel cryopreserved for 5 and 15 days exhibited no significant different in apoptosis rates relative to the control group. Extending the cryopreservation time to 45 days resulted in significantly increased and decreased apoptosis and volume retention rates of SVF-gel, respectively. SVF-gel grafts cryopreserved for 5 and 15 days exhibited no significant differences from those in the control group, although their weights and volumes still fluctuated. Extending the cryopreservation time to 45 days resulted in significantly decreased retention rates of the grafts. Histologically, extending freezing time resulted in a gradual decline in the graft's health adipose tissue, as well as decreased angiogenesis, and connective tissue hyperplasia. CONCLUSION: Simple freezing of SVF-gel at - 20 °C conferred them with sufficient cell viability. Notably, short-term cryopreservation did not significantly increase the apoptosis rate, and it still had a certain regeneration after transplantation. However, prolonging freezing time to 45 days resulted in increased apoptosis rate and worsened transplantation effect. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Identification and Validation of a Novel Model: Predicting Short-Term Complications After Local Flap Surgery for Skin Tumor Removal.

BACKGROUND The aim was to analyze the risk factors for the occurrence of complications after local flap transfer and to construct a simple prediction model to help surgeons in the perioperative screening of high-risk patients. MATERIAL AND METHODS Short-term complications were defined as any postoperative infection, dehiscence, bleeding, subcutaneous effusion, fat liquefaction, arteriovenous crisis, and tissue necrosis that required medical consultation or intervention. To explore 16 factors influencing short-term complications after local flap transfer, least absolute shrinkage and selection operator (LASSO) logistic regression was used to reduce the dimensionality of the data and to screen for predictors. Independent risk factors affecting the development of complications after local flap transfer were analyzed using logistic multiple regression models. The consistency (C-)index, receiver operating characteristic (ROC) curves, and calibration curves were used to check the model's discrimination and calibration. Decision curve analysis (DCA) curves were used to evaluate the clinical applicability of this model, and internal validation was assessed using bootstrap validation. RESULTS The C-index of the nomogram model to predict short-term complications after local flap transfer was 0.763 (95% CI: 0.702-0.824), the area under the ROC curve was 0.763, and the internal validation C-index was 0.747. The calibration curve showed good agreement between observed and predicted values, and the DCA showed the model can benefit patients. CONCLUSIONS The model identified the relevant factors influencing short-term complications after local flap transfer, facilitating the identification and targeted intervention of patients at high risk of flap complications after surgery.

i6mA-Vote: Cross-Species Identification of DNA N6-Methyladenine Sites in Plant Genomes Based on Ensemble Learning With Voting.

DNA N6-Methyladenine (6mA) is a common epigenetic modification, which plays some significant roles in the growth and development of plants. It is crucial to identify 6mA sites for elucidating the functions of 6mA. In this article, a novel model named i6mA-vote is developed to predict 6mA sites of plants. Firstly, DNA sequences were coded into six feature vectors with diverse strategies based on density, physicochemical properties, and position of nucleotides, respectively. To find the best coding strategy, the feature vectors were compared on several machine learning classifiers. The results suggested that the position of nucleotides has a significant positive effect on 6mA sites identification. Thus, the dinucleotide one-hot strategy which can describe position characteristics of nucleotides well was employed to extract DNA features in our method. Secondly, DNA sequences of Rosaceae were divided into a training dataset and a test dataset randomly. Finally, i6mA-vote was constructed by combining five different base-classifiers under a majority voting strategy and trained on the Rosaceae training dataset. The i6mA-vote was evaluated on the task of predicting 6mA sites from the genome of the Rosaceae, Rice, and Arabidopsis separately. In Rosaceae, the performances of i6mA-vote were 0.955 on accuracy (ACC), 0.909 on Matthew correlation coefficients (MCC), 0.955 on sensitivity (SN), and 0.954 on specificity (SP). Those indicators, in the order of ACC, MCC, SN, SP, were 0.882, 0.774, 0.961, and 0.803 on Rice while they were 0.798, 0.617, 0.666, and 0.929 on Arabidopsis. According to the indicators, our method was effectiveness and better than other concerned methods. The results also illustrated that i6mA-vote does not only well in 6mA sites prediction of intraspecies but also interspecies plants. Moreover, it can be seen that the specificity is distinctly lower than the sensitivity in Rice while it is just the opposite in Arabidopsis. It may be resulted from sequence similarity among Rosaceae, Rice and Arabidopsis.

i4mC-EL: Identifying DNA N4-Methylcytosine Sites in the Mouse Genome Using Ensemble Learning.

As one of important epigenetic modifications, DNA N4-methylcytosine (4mC) plays a crucial role in controlling gene replication, expression, cell cycle, DNA replication, and differentiation. The accurate identification of 4mC sites is necessary to understand biological functions. In the paper, we use ensemble learning to develop a model named i4mC-EL to identify 4mC sites in the mouse genome. Firstly, a multifeature encoding scheme consisting of Kmer and EIIP was adopted to describe the DNA sequences. Secondly, on the basis of the multifeature encoding scheme, we developed a stacked ensemble model, in which four machine learning algorithms, namely, BayesNet, NaiveBayes, LibSVM, and Voted Perceptron, were utilized to implement an ensemble of base classifiers that produce intermediate results as input of the metaclassifier, Logistic. The experimental results on the independent test dataset demonstrate that the overall rate of predictive accurate of i4mC-EL is 82.19%, which is better than the existing methods. The user-friendly website implementing i4mC-EL can be accessed freely at the following.

The Modified Suction-curettage Technique for Minimally Invasive Treatment of Axillary Osmidrosis.

BACKGROUND: Axillary osmidrosis (AO) is a common and nonnegligible disease, the treatment of which is currently lacking a consensus. AIMS: The aim of this study was to introduce a modified suction-assisted technique as a safer and more efficient surgical procedure. METHODS: This retrospective clinical study included 80 patients who recieved a modified suction-curettage procedure (group A) or a subcutaneous gland excision procedure (group B). Intraoperative assessment (endoscopy and pathological biopsy) and postoperative assessment (complications, therapeutic effect, and satisfaction) were performed for both groups. RESULTS: The endoscopy and pathological biopsy results demonstrated that the modified suction-curettage technique could remove the apocrine gland efficiently. Compared with group B, a lower complication rate (long-term, 5.00%; P=0.014, and short-term, 11.10%; P=0.001) and higher patient satisfaction (98.00%, P=0.012) were observed in group A. CONCLUSION: The modified suction-curettage procedure is an effective and safe treatment for axillary osmidrosis.

Influence of combined epidural anesthesia on cognitive function, inflammation and stress response in elderly liver cancer patients undergoing surgery.

Effects of combined epidural anesthesia on the cognitive function, inflammation and stress response in the elderly liver cancer patients undergoing surgery were explored. Elderly liver cancer patients (n=100) undergoing surgery in the Second Affiliated Hospital of Dalian Medical University from January 2015 to December 2018 were enrolled and randomly divided into observation group (n=50) and control group (n=50). In control group only conventional anesthesia was performed using 2 µg/kg fentanyl, 1.5 mg/kg propofol and 0.2 mg/kg atracurium, in addition to the procedures in the control group, combined epidural anesthesia was administered using 0.5% bupivacaine for 15 sec and maintained via 0.25% bupivacaine in the observation group. The anesthetic effect was observed and the arterial oxygen saturation (SaO2), heart rate (HR), mean arterial pressure (MAP) and mini-mental state examination (MMSE) and cognitive function scores by cognitive abilities screening instrument (CASI) were evaluated in the patients, and their blood was drawn to detect the inflammatory factors interleukin (IL)-6, IL-1 and tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), norepinephrine and epinephrine. The observation group exhibited a better anesthetic effect and obviously smaller decreases in the SaO2 and MAP and increase in HR than the control group (P<0.05). The MMSE and CASI scores, and the content of IL-1, IL-6, TNF-α, MDA, CAT, norepinephrine and epinephrine in the observation group was obviously lower than that in the control group (P<0.05), while the content of SOD was evidently higher than that in the control group (P<0.05). Overall postoperative conditions in the observation group was superior to the control group (P<0.05), with the incidence rate of cognitive disorder lower than that in the control group (P<0.05). Combined epidural anesthesia dramatically improves the postoperative conditions and cognitive function and relieve inflammatory and stress responses in the patients with a better anesthetic effect, thus holding promise for application.

Comparative transcriptomic analysis provides insights into the development of a Salvia splendens Ker-Gawler mutant, SX919M.

Salvia splendens is a perennial, ornamental herbaceous flower that is widely cultivated as a bedding plant in gardens. The development of novel S. splendens cultivars and investigating the relevant molecular mechanisms are of great significance. In this study, RNA-sequencing and real-time PCR methods were used to analyze the possible molecular mechanism of S. splendens mutant, SX919M. From the wild-type S. splendens 919CK, we firstly selected a natural mutant, SX919M, which displayed multiple branches, clustered spheroids, and radial symmetrical inflorescence with higher numbers of calyces, ovules, stamens, and perianth tubes. Further, the RNA-seq was used to identify the differentially expressed genes (DEGs) in the mutant which included a total of 3568 upregulated and 3290 downregulated unigenes. We further observed that the indole alkaloid biosynthesis pathway showed the highest DEG enrichment, which was supported by a significant increase in the IAA content in mutant SX919M. In addition, we validated three DEGs, namely, CL2200.Contig2_All encoding methyl IAA esterase, CL12462.Contig1_All and CL12462.Contig2_All, which encoded strictosidine synthase, upregulated in mutant SX919M. We selected a novel S. splendens germplasm SX919M with a high ornamental value and determined that the upregulation of IAA biogenesis may be associated with its development.

[Application of delayed skin-stretching device in treatment of skin and soft tissue defects].

Objective: To summarize the effectiveness of delayed skin-stretching device in treatment of skin and soft tissue defects. Methods: Between December 2014 and December 2016, 10 cases of skin and soft tissue defects were treated with delayed skin-stretching device. There were 6 males and 4 females with an average age of 53 years (range, 42-64 years). The skin and soft tissue defects were caused by acute trauma in 6 cases. The incision could not be closed directly after making incisions because of osseous fascia syndrome in 3 cases. The skin soft tissue defect caused after huge carbuncle incision and drainage in 1 case. The defect located at thigh in 4 cases, lower leg in 3 cases, upper arm in 2 cases, back in 1 case. The defect area ranged from 10 cm×4 cm to 22 cm×12 cm. Pinch test was performed on the wound margin, which confirmed that the wound could not be closed directly. Results: Tension blisters were found in 3 cases during traction, and no complications such as impaired blood circulation or skin necrosis occurred in all cases. Skin defects closed directly after continuously stretching for 7-18 days. No skin graft or free flap repair was performed in all patients. The wound healed well after operation. All the 10 patients were followed up 5-8 months (mean, 6.5 months). There was no necrosis around the wound margin and the scar was linear. The sensation and function were not affected. Conclusion: Delayed skin-stretching device is an effective method to treat skin and soft tissue defects, which has the advantages of simple operation, lower risk of operation, less complications, and reliable effectiveness.

High-quality assembly of the reference genome for scarlet sage, Salvia splendens, an economically important ornamental plant.

Background: Salvia splendens Ker-Gawler, scarlet or tropical sage, is a tender herbaceous perennial widely introduced and seen in public gardens all over the world. With few molecular resources, breeding is still restricted to traditional phenotypic selection, and the genetic mechanisms underlying phenotypic variation remain unknown. Hence, a high-quality reference genome will be very valuable for marker-assisted breeding, genome editing, and molecular genetics. Findings: We generated 66 Gb and 37 Gb of raw DNA sequences, respectively, from whole-genome sequencing of a largely homozygous scarlet sage inbred line using Pacific Biosciences (PacBio) single-molecule real-time and Illumina HiSeq sequencing platforms. The PacBio de novo assembly yielded a final genome with a scaffold N50 size of 3.12 Mb and a total length of 808 Mb. The repetitive sequences identified accounted for 57.52% of the genome sequence, and 54,008 protein-coding genes were predicted collectively with ab initio and homology-based gene prediction from the masked genome. The divergence time between S. splendens and Salvia miltiorrhiza was estimated at 28.21 million years ago (Mya). Moreover, 3,797 species-specific genes and 1,187 expanded gene families were identified for the scarlet sage genome. Conclusions: We provide the first genome sequence and gene annotation for the scarlet sage. The availability of these resources will be of great importance for further breeding strategies, genome editing, and comparative genomics among related species.

Soybean isoflavone treatment induces osteoblast differentiation and proliferation by regulating analysis of Wnt/ß-catenin pathway.

It has been reported that soybean isoflavones (SI) have anti-bone resorptive activity in vivo. However, little is known about the cellular and molecular mechanisms of this effect. In this study, we investigated the effects of SI on osteoblast differentiation and proliferation. Results showed that SI promoted osteoblast proliferation and differentiation which could be marked as the formation of hydroxyapatite crystals or extracellular matrix and increased ALP level. SI treatment markedly decreased RANKL levels and increased OPG levels in osteoblast cells. SI resulted in activation of the Wnt/ß-catenin pathway. These results demonstrate that SI may be a useful remedy for the treatment of osteoporosis.

Mapping QTL controlling maize deep-seeding tolerance-related traits and confirmation of a major QTL for mesocotyl length.

Deep-seeding tolerant seeds can emerge from deep soil where the moisture is suitable for seed germination. Breeding deep-seeding tolerant cultivars is becoming increasingly important in arid and semi-arid regions. To dissect the quantitative trait loci (QTL) controlling deep-seeding tolerance traits, we selected a tolerant maize inbred line 3681-4 and crossed it with the elite inbred line-X178 to generate an F(2) population and the derivative F(2:3) families. A molecular linkage map composed of 179 molecular markers was constructed, and 25 QTL were detected including 10 QTL for sowing at 10 cm depth and 15 QTL for sowing at 20 cm depth. The QTL analysis results confirmed that deep-seeding tolerance was mainly caused by mesocotyl elongation and also revealed considerable overlap among QTL for different traits. To confirm a major QTL on chromosome 10 for mesocotyl length measured at 20 cm depth, we selected and self-pollinated a BC(3)F(2) plant that was heterozygous at the markers around the target QTL and homozygous at other QTL to generate a BC(3)F(3) population. We found that this QTL explained more phenotypic variance in the BC(3)F(3) population than that in the F(2) population, which laid the foundation for fine mapping and NIL (near-isogenic line) construction.