Search

Bottle gourd [Lagenaria siceraria (Mol.) Standl.] is one of the most common horticultural and medicinal crops. Genetic transformation is a rate-limiting step in unraveling the function of genes and molecular breeding in this non-model crop. In this study, we established an optimized protocol for transformation using a seed germination pouch as a growth carrier and developed bottle gourd composite plants. Overground explants were inoculated with Agrobacterium rhizogenes strain R1000, which harbored a binary vector pBI121 carrying the GUS reporter gene. The integration of GUS into transgenic roots was confirmed via reverse transcription-polymerase chain reaction and histochemical staining. Furthermore, we screened for desirable genotypes from 14 bottle gourd germplasm collections based on the number of induced roots, transformation efficiency, and induction rate, and “Changguang” (transformation efficiency, approximately 36.8%) was selected as the genotype suitable for hairy root induction in bottle gourd. The optimized protocol presented here is a rapid, suitable, and efficient tool for studying the function of genes and root biology of bottle gourd in the future.

Intracerebral hemorrhage (ICH) is one of the most lethal stroke types and lacks effective therapeutic regimens. Recently, evidence has suggested the involvement of the ferroptosis inhibitor ferrostatin-1 (Fer-1) in the pathophysiological process of ICH. In this study, we examined the underlying mechanism.We induced an in vitro apoptosis model in organotypic hippocampal slice (OHS) using hemoglobin (Hb) and an in vivo ICH model using collagenase. OHSs were treated with MK-801, Fer-1, glutamate, and Hb to assess the impacts of Fer-1 on neuron apoptosis, glutathione peroxidase-4 activity, reactive oxygen species production, inflammation-related factors, expression of M1 markers and M2 markers, and the phagocytic function of microglial cells in vitro. Then, ICH mice were treated with Fer-1 and ruxolitinib to evaluate the effects of Fer-1-orchestrating janus kinase 1/signal transducer and activator of transcription 6 pathway on neurological function, brain water content, hematoma volume, the anti-inflammatory factor, M1 and M2 markers, and the phagocytic function of microglial cells in vivo.Hb or glutamate facilitated glutathione peroxidase dysfunction, reactive oxygen species production, and neuronal apoptosis in OHSs, which was nullified by Fer-1. Fer-1 polarized microglial cells to the M2 phenotype, enhanced their phagocytic function, and prevented inflammation in Hb-induced OHSs. In the ICH mouse model, Fer-1 was found to improve neurological function and promote hematoma absorption. In addition, Fer-1 activated the Fer-1-orchestrating janus kinase 1/signal transducer and activator of transcription 6 pathway, which accelerated microglial M2 polarization, enhanced the phagocytic function of microglial cells, and restrained inflammation in ICH mice.Overall, our findings suggest that Fer-1 may be a novel mechanism underlying microglial M2 polarization and inflammation after ICH.

Background: For the majority of the patients, surgical treatment of primary infantile exotropia (PIE) with large exodeviation angels of more than 50 prism diopters (PD) is not satisfactory. We evaluate the effectiveness of augmented lateral rectus (LR) muscle recession in treatment of congenital exotropia with large deviation angles. Methods: A retrospective analysis was performed for 25 patients with congenital exotropia who had at least 60 prism diopters (PD). Augmented LR recession was performed using a hemi hang-back technique. The ocular alignment, ocular motility and binocular vision were evaluated pre- and postoperatively. Results: There were 14 male and 11 female affected individuals with congenital exotropia included in this study, with a mean age of (3.92±2.02) years (range, 1 - 7 years). The average exodeviation angle was (68.0±8.78) PD (range, 60 - 80 PD) preoperatively. All patients underwent bilateral LR recession, of whom 3 patients underwent bilateral inferior oblique muscle transposition in the same operation. The mean surgical dosage was (11.08±1.19) mm for each lateral rectus muclse. All patients did not have abduction deficiency after a large amount of lateral rectus recession. 21 of 25 patients (84.0%) acquired orthophoria at the primary gaze position at the final visit. 13 patients obtained binocular vision. Conclusions: Augmented bilateral LR recession is an effective and safe surgical procedure for treatment of congenital exotropia with large deviation angles.

Background Head and Neck Squamous Cell Carcinoma is a malignant tumor with high morbidity and mortality. The MMP family plays an important role in tumor invasion and metastasis. However, the mechanistic value of the MMP family as a therapeutic target and prognostic biomarker in HNSC has not been fully elucidated. Methods Oncomine, UALCAN, GEPIA, cBioportal, GeneMANIA, STRING, DAVID6.8, TRRUST, TIMER and Linkedomics were used for analysis. Results The mRNA expression levels of MMP1, MMP3, ILF3, MMP7, MMP9, MMP10, MMP11, MMP12, MMP13 and MMP16 were higher in HNSC than those in normal tissues, while the mRNA expression level of MMP15 was reduced. The relative expression levels of MMP1 and MMP14 were the highest in HNSC tissues. A significant correlation was found between the expression of MMP3, MMP11, MMP25 and the pathological stage of HNSC patients. There was no significant associations between all the MMP family members expression levels and DFS. Increased mRNA levels of MMP1, MMP8 and MMP25 were significantly associated with OS. In addition, we investigated the genetic changes of the MMP family in HNSC and found that all the MMP family members had genetic changes, most of which were amplification and depth loss. In the analysis of neighbor gene network and protein interaction, we found that the MMP family interacted with 25 neighboring genes, except for ILF3, MMP19, MMP20, MMP21, MMP23B, MMP27 and MMP28, other MMP proteins interacted with each other. Functional enrichment analysis showed that the MMP family could be present in the extracellular matrix, regulate peptidase activity, and participate in the catabolism of collagen. Meanwhile, we identified the transcription factor targets and kinase targets of the MMP family and found that ATM and ATR were the two most common kinase targets in the MMP family. We also found a significant correlation between the MMP family expression and immune cell infiltration. Cox proportional risk model analysis showed that macrophages, MMP14, MMP16, and MMP19 were significantly associated with clinical outcomes in HNSC patients. Conclusion The MMP family might serve as therapeutic target and prognostic biomarker in HNSC.

Objective To investigate the role of standard patients (SPs) and examiners as assessors for scoring in the dental objective structured clinical examination (OSCE) system and to evaluate the scoring differences between them. Methods We developed the doctor-patient communication and clinical examination station in the OSCE system. The examination time of this station was 10 min, and the examination institution wrote the script and recruited SPs. A total of 146 examinees who received standardized resident training at the Nanjing Stomatological Hospital, Medical School of Nanjing University between 2018 and 2021 were assessed. They were scored by SPs and examiners according to the same scoring rubrics. Subsequently, the SPSS software was used to analyze the examination results of different assessors and evaluate the consistency. Results The average score of all examinees provided by SPs and examiners was 90.45 ± 3.52 and 91.53 ± 4.13, respectively. The consistency analysis showed that the intraclass correlation coefficient was 0.718, which was indicative of medium consistency. Conclusion Our findings showed that SPs could be used directly as assessors, as they could provide a simulated and realistic clinical setting and create favorable conditions for comprehensive competence training and improvement for medical students.

Fault diagnosis plays an important role in the measuring of the fault tolerance of an interconnection network, which is of great value in the design and maintenance of large-scale multiprocessor systems. As a classical variant of the hypercube, the Balanced Hypercube, denoted by BHn(n $$\ge$$ 1), has drawn a lot of research attention, and its $$g$$ -extra diagnosability has been studied to improve the network diagnostic ability. However, the current literatures on $$g$$ -extra diagnosability of BHn under the PMC model only cover the cases of $$g < 6$$ , and what’s more, seldom involve its $$g$$ -extra diagnosability under the MM* model, which is a great limitation on the research of BHn diagnosability. In this paper, the upper and lower bounds of the $$g$$ -extra diagnosability of the balanced hypercube are proved, respectively, based on the $$g$$ -extra connectivity by the contradiction method, and finally, the $$g$$ -extra diagnosability of BHn for $$2 \le g \le 2n - 1$$ under the PMC and MM* model is obtained, i.e., $$2\left[ {\left( {n - 2} \right)\lceil\frac{g - 1}{2}\rceil + n} \right] + g$$ . In addition, as a special case, the $$g$$ -extra diagnosability of the balanced hypercube for $$g = 2n$$ is proved to be $$2^{2n - 1} - 1$$ under the PMC and MM* model. In the end, simulation experiments are conducted to verify the effectiveness of our proposed theories. The conclusion of this paper has certain theory and application value for the research of BHn fault diagnosis.

Objective We aimed to investigate the effect of differentially methylated genes and chronic childhood stress on the development of depressive symptoms in Chinese adolescents, as well as to test whether methylation at baseline can be used as a predictor of remission at follow-up after six weeks of treatment. Methods After recruiting 87 MDD patients and 53 healthy controls, we compared demographic and baseline clinical characteristics. The Childhood Chronic Stress Questionnaire was used to assess stress caused by early-life events. MDD patients underwent six weeks of treatment, and response to treatment was assessed using the Beck Depression Inventory-II. In addition, four MDD patients and five controls were randomly chosen for genome-wide methylation analysis. Results The gene RPS6KA5 showed significant methylation differences between the two groups. Severity of chronic childhood stress was significantly associated with increased risk of depression in adolescents, but not with treatment response. Baseline RPS6KA5 methylation can predict remission after six weeks of treatment. We did not observe any interaction between RPS6KA5 methylation and chronic childhood stress. Conclusions Our results suggest that RPS6KA5 methylation can be used as a predictor of response to treatment in adolescent MDD patients. Here we offer new evidence for the role of epigenetics in early response to treatment of depression. Trial registration ChiCTR, ChiCTR2000033402, 31/05/2020, http://www.chictr.org.cn/index.aspx

Mutations in the FERM domain containing 7 (FRMD7) gene have been proven to be responsible for infantile nystagmus (IN). The purpose of this study is to investigate FRMD7 gene mutations in patients with IN, and to evaluate the nystagmus intensity among patients with and without FRMD7 mutations. The affected males were subdivided into three groups according to whether or not having FRMD7 mutations and the types of mutations. Fifty-two mutations were detected in FRMD7 in 56 pedigrees and 34 sporadic patients with IN, including 28 novel and 24 previous reported mutations. The novel identified mutations further expand the spectrum of FRMD7 mutations. The parameters of nystagmus intensity and the patients’ best corrected visual acuity were not statistically different among the patients with and without identified FRMD7 mutations, and also not different among patients with different mutant types. The FERM-C domain, whose amino acids are encoded by exons 7, 8 and 9, could be the harbor region for most mutations. Loss-of-function is suggested to be the common molecular mechanism for the X-linked infantile nystagmus.

A capillary-based SERS sensor was fabricated for ultrasensitive and selective detection of Hg2+ in water. Au@Ag core–shell NPs embedded with 4-mercaptobenzoic acid (4-MBA) (Au@4-MBA@Ag) were prepared by a seed growth method and fixed on the inner wall of the glass capillary to obtain the sensor. Owing to the amalgamation between Ag and Hg, the capillary-based SERS sensor can specifically recognize the reduced Hg2+ without any recognition element, and the resulted Ag/Hg amalgam can weaken the SERS activity of Ag shell; thus, the SERS intensity of the embedded 4-MBA at 1075 cm−1 gradually decreased with the increase of Hg2+ concentration. Under the optimum condition, the fabricated sensor can sensitively determine Hg2+ in water with a limit of detection (LOD) as low as 0.03 nM. The capillary-based SERS sensor offers the advantages of simple preparation, superior stability, and high selectivity, which is promising for rapid and on-site detection of Hg2+ in water combined with a portable Raman device.

Few plant species used for revegetation grow well in multi-metal-contaminated soils. Vetiver grass (Vetiveria zizanioides) is known to be tolerant of heavy metals. Vetiver has been reported to be effective for revegetation and heavy metal phytoextraction by applying targeted amendments due to its large biomass. In this study, a greenhouse vetiver pot experiment and soil incubation were performed to investigate the growth and Cd, Cr, Cu, Pb, and Zn uptake of vetiver grown in multi-metal-contaminated soils treated with a CaO-activated Si-based slag amendment (0, 0.5, 1.0, and 2.0% w/w). The results showed that the effects of slag amendment on plant growth and heavy metal uptake and distribution were dependent on the amendment dosages and metal species. Although vetiver could grow in contaminated soils, its growth was obviously inhibited. The slag amendment enhanced the vetiver growth and the highest biomass (2.62-fold over the control) was determined at a 1.0% amendment rate. The slag amendment improved plant growth by alleviating the toxicity of heavy metals in plants. This result was mainly attributed to the increases in soil pH and citric acid-extractable Si caused by alkaline amendment. The results suggest that vetiver can be applied to remediate multi-metal-contaminated soils in conjunction with the application of CaO-activated Si-based slag amendment.

Researches have revealed the vital roles of the generated immune environment via the response of immune cells growing on biomaterial surfaces in the bone healing process. HAS and novel constructed microgrooved patterns of HAS (HAS-G) are widely used as biocompatible ceramic, especially as a mimic of the natural bone matrix. However, it is unclear whether osteoimmune response induced by HAS and HAS-G affects the osteogenic differentiation of bone marrow stromal cells (BMSCs). RAW264.7 cells were seeded on different surface of materials and cytokines released by macrophages were detected by enzyme-linked immunosorbent assay. The cell viability and mitochondrial function of macrophages seeded on different surface of materials were detected. Then, the effects of modified inflammatory microenvironment by macrophages on osteogenesis of BMSCs were measured by performing ALP staining, Alizarin Red S staining, and western blot. We confirmed that HAS-G is more favorable for RAW cell attaching and subsequently regulated the expression and release of cytokines/chemokines. Decrease in interleukin-6 (IL-6) release was further confirmed for contributing significantly to improve mitochondrial function in RAW cells. HAS-G-conditioned medium promoted osteogenic differentiation in BMSCs and was reversed by IL-6 addition. Decrease in IL-6 contributes to downregulation of miR-214 and subsequently upregulated p38/JNK pathway, which is potentially contributes to osteogenic promotion by HAS-G. This study is the first report to reveal the effects of HAS-G on osteogenesis via immune response, which could lead to a new insight into novel material for the advantage of biomaterials for tissue engineering applications.

BackgroundThe incidence of ischemic stroke in the context of vascular disease is high, and the expression of growth-associated protein-43 (GAP43) increases when neurons are damaged or stimulated, especially in a rat model of middle cerebral artery occlusion/reperfusion (MCAO/R).ExperimentaldesignWe bioengineered neuron-targeting exosomes (Exo) conjugated to a monoclonal antibody against GAP43 (mAb GAP43) to promote the targeted delivery of quercetin (Que) to ischemic neurons with high GAP43 expression and investigated the ability of Exo to treat cerebral ischemia by scavenging reactive oxygen species (ROS).ResultsOur results suggested that Que loaded mAb GAP43 conjugated exosomes (Que/mAb GAP43-Exo) can specifically target damaged neurons through the interaction between Exo-delivered mAb GAP43 and GAP43 expressed in damaged neurons and improve survival of neurons by inhibiting ROS production through the activation of the Nrf2/HO-1 pathway. The brain infarct volume is smaller, and neurological recovery is more markedly improved following Que/mAb GAP43-Exo treatment than following free Que or Que-carrying exosome (Que-Exo) treatment in a rat induced by MCAO/R.ConclusionsQue/mAb GAP43-Exo may serve a promising dual targeting and therapeutic drug delivery system for alleviating cerebral ischemia/reperfusion injury.

With the rapid development of e-commerce, the backlog of distribution orders, insufficient logistics capacity and other issues are becoming more and more serious. It is very significant for e-commerce platforms and logistics enterprises to clarify the demand of logistics. To meet this need, a forecasting indicator system of Guangdong logistics demand was constructed from the perspective of e-commerce. The GM (1, 1) model and Back Propagation (BP) neural network model were used to simulate and forecast the logistics demand of Guangdong province from 2000 to 2019. The results show that the Guangdong logistics demand forecasting indicator system has good applicability. Compared with the GM (1, 1) model, the BP neural network model has smaller prediction error and more stable prediction results. Based on the results of the study, it is the recommendation of the authors that e-commerce platforms and logistics enterprises should pay attention to the prediction of regional logistics demand, choose scientific forecasting methods, and encourage the implementation of new distribution modes.

This study investigated the effects of alkaline silicon-based amendment (A) and incorporated with biochar (B) on the growth, heavy metal translocation, and accumulation of vetiver grass (Vetiveria zizanioides) grown in multi-metal-contaminated soils. Soil incubation for 6 months and pot experiments for 6 months were performed to investigate the growth and the translocation factor (TF) and bioaccumulation factor (BCF) values of heavy metals in vetiver grown in As-, Cd-, Cr-, and Pb-contaminated soils amended with A (0.5%) and AB (0.5% A; 1.5% B). The vetiver could grow in multi-metal-contaminated soils due to the restriction of metals translocation in vetiver. Vetiver could be the plant for phytostabilization of As, Cr, Cd, and Pb because As, Cr, Cd, and Pb accumulated by the vetiver were largely retained in the roots, as the TF values for As, Cr, Cd, and Pb were

Vascular endothelial growth factor B (VEGF-B) was discovered a long time ago. However, its role in hyperglycemia- and VEGF-A inhibition-induced retinal apoptosis remains unknown thus far. Yet, drugs that can block VEGF-B are being used to treat patients with diabetic retinopathy and other ocular neovascular diseases. It is therefore urgent to have a better understanding of the function of VEGF-B in these pathologies. Here, we report that both streptozotocin (STZ)-induced diabetes in rats and Macugen intravitreal injection in mice leads to retinal apoptosis in retinal ganglion cell and outer nuclear layers respectively. Importantly, VEGF-B treatment by intravitreal injection markedly reduced retinal apoptosis in both models. We further reveal that VEGF-B and its receptors, vascular endothelial growth factor 1 (VEGFR1) and neuropilin 1 (NP1), are abundantly expressed in rat retinae and choroids and are upregulated by high glucose with concomitant activation of Akt and Erk. These data highlight an important function of VEGF-B in protecting retinal cells from apoptosis induced by hyperglycemia and VEGF-A inhibition. VEGF-B may therefore have a therapeutic potential in treating various retinal degenerative diseases, and modulation of VEGF-B activity in the eye needs careful consideration.

CD4(+)CD28(null) T cells are present in increased numbers in the peripheral blood of patients with acute coronary syndrome. However, the triggers of expansion of these cells are unclear. Susceptibility to coronary heart disease (CHD) is strongly associated with alleles of human leukocyte antigen (HLA), but it is not equally strong in different human populations. The objective of the study was to investigate association between CD4(+)CD28(null) T cells and HLA-DRB1 alleles. The HLA alleles were determined by polymerase chain reaction with sequence-specific primers (PCR-SSP) method, in a group of CHD patients and control subjects from the same area. The number of CD4(+)CD28(null) T cells was measured using the flow cytometry technique. The HLA-DRB1*01 (RR = 4.705, P0.005) and DRB1*04 (RR = 3.554, P0.005) alleles showed the strongest association with CHD in the Chinese population, and increased numbers of CD4(+)CD28(null) T cells were found in association with HLA-DRB1*04 (17.1%) and DRB*01 (12.9%), while decreased numbers of CD4(+)CD28(null) T cells were present in subjects with DRB1*15 (0.8%). CHD in Chinese population is strongly associated with HLA-DRB1*01 and DRB1*04 haplotypes, and formation of CD4(+)CD28(null) T cells was related to HLA-DRB1*01, DRB1*04, and DRB1*15 alleles.

We demonstrate that single-nucleotide variations in a DNA sequence can be detected using capillary electrophoresis (CE) and molecular beacons (MBs). In this method, the region surrounding the site of a nucleotide variation was amplified in a polymerase chain reaction, then hybridize PCR products with each of MBs. The sequences of the PCR products are different at the site of 2,044 in exon of interleukin (IL)-13 which to be identified. Through denaturation, the PCR product became single strand and hybridized with the completely complementary MB. The MB-target duplexes were separated using CE and solution-based fluorescence techniques. The results show that in each reaction a fluorescent response was elicited from the molecular beacon which was perfectly complementary to the amplified DNA, but not from the other MB whose probe sequence mismatched the target sequence. The method of CE based on MBs is able to identify single-nucleotide variations in a DNA sequence and can discriminate the genotyping of the SNP between the homo- and heteroduplexes of DNA fragments.

WNT5A is one of the most highly investigated non-canonical Wnt ligands and is involved in the embryonic heart development, especially in formation of the cardiac conotruncal region by regulating the migration and differentiation of cardiac neural crest (CNC) and second heart field (SHF) cells. No study to date has comprehensively characterized the WNT5A regulatory variants in patients with congenital heart malformations (CHMs). The association between regulatory variants of the WNT5A gene and CHMs was examined in case-control association study in 1,210 CHMs and 798 controls. Individuals carrying a homozygous genotype CC (rs524153) or GG (rs504849) had a similarly reduced risk of conotruncal malformations. The homozygous genotypes (CC for rs524153 and GG for rs504849) were associated with a lower WNT5A transcriptional level compared with the transcriptional level of those with wild-type genotypes. Further functional analysis revealed that an additional upstream single nucleotide polymorphisms (SNP) rs371954924 (–5244GCCA > CC) in a linkage disequilibrium (LD) block with the above genotyped SNPs decreased WNT5A expression through the attenuated binding affinity with the transcription factor SOX9. This is the first demonstration that genetic variants in the regulatory regions of WNT5A play a vital role in sporadic conotruncal malformations susceptibility through the changeable expression of the WNT5A gene.

The isothermal oxidation behavior of a single-crystal (SC) Ni-base superalloy was studied over the temperature range from 900 to 1100 °C and analyzed by OM, TGA, XRD, EDX, SEM and EPMA. The alloy obeyed subparabolic rate law during oxidation at 900 and 1000 °C, whereas the alloy showed parabolic rate law at 1100 °C exposure. The variations in the chemical composition due to segregations, which resulted from the solidification process, led to the formation of different kinds of oxide scale on the dendritic and interdendritic area during oxidation at 900 and 1000 °C, while the alloy exhibited relatively uniform oxidation at 1100 °C. The growth mechanism of the scale was discussed.