Overexpression of HbGRF4 or HbGRF4-HbGIF1 Chimera Improves the Efficiency of Somatic Embryogenesis in Hevea brasiliensis.

Transgenic technology is a crucial tool for gene functional analysis and targeted genetic modification in the para rubber tree (Hevea brasiliensis). However, low efficiency of plant regeneration via somatic embryogenesis remains a bottleneck of successful genetic transformation in H. brasiliensis. Enhancing expression of GROWTH-REGULATING FACTOR 4 (GRF4)-GRF-INTERACTING FACTOR 1 (GIF1) has been reported to significantly improve shoot and embryo regeneration in multiple crops. Here, we identified endogenous HbGRF4 and HbGIF1 from the rubber clone Reyan7-33-97, the expressions of which dramatically increased along with somatic embryo (SE) production. Intriguingly, overexpression of HbGRF4 or HbGRF4-HbGIF1 markedly enhanced the efficiency of embryogenesis in two H. brasiliensis callus lines with contrasting rates of SE production. Transcriptional profiling revealed that the genes involved in jasmonic acid response were up-regulated, whereas those in ethylene biosynthesis and response as well as the S-adenosylmethionine-dependent methyltransferase activity were down-regulated in HbGRF4- and HbGRF4-HbGIF1-overexpressing H. brasiliensis embryos. These findings open up a new avenue for improving SE production in rubber tree, and help to unravel the underlying mechanisms of HbGRF4-enhanced somatic embryogenesis.

Benign Brenner tumor of the ovary: two-dimensional and contrast-enhanced ultrasound features-a retrospective study from a single center.

Objective: Benign Brenner tumor (BBT) is a rare ovarian tumor, and there are few discrete reports about its manifestation in an ultrasound. This study sought to investigate the two-dimensional (2D) and contrast-enhanced ultrasound (CEUS) features of this entity. Methods: This is a retrospective single-center study. The clinical manifestations, laboratory examination, and ultrasound data of 25 female patients with BBT were confirmed by pathology when they underwent 2D and/or CEUS examination at Ningbo First Hospital from January 2012 to June 2023. The ultrasound findings of the patients were analyzed using the terminology of the International Organization for the Analysis of Ovarian Tumor and were read by two senior sonographers who reached an agreement. Results: Among the all 25 patients, most of them were unilateral, and only one patient was bilateral. Thus, 26 lesions were found: 44.0% (11/25) were in the left and 52.0% (13/25) were in the right. Moreover, 53.84% (14/26) were solid lesions, 15.38% (4/26) were mixed lesions, and 26.92% (7/26) were cystic lesions. Among the solid-type patients, 42.85% (6/14) of the cases were with calcification. Upon laboratory examination, 12.0% (3/25) of the patients had high carbohydrate antigen 125 (CA-125) level, and 19.04% (4/21) of the patients had an elevated carbohydrate antigen724 (CA-724) level in the serum tumor markers. In the hormone test, 14.28% (3/21) were found to have a high postmenopausal estrogen level and 14.28%(3/21) were found to have a high level of follicle-stimulating hormone (FSH). One patient with complex manifestations and three with solid manifestations were examined by CEUS to observe the microcirculation perfusion of the tumor. One with solid and cystic separation was rapidly hyperenhanced and cleared, and the filling subsided faster than the uterus. The postoperative pathological diagnosis was benign Brenner tumor with mucinous cystadenoma. The other three cases were solid adnexal lesions, which showed isoenhancement on CEUS and disappeared slowly, synchronizing with the uterus. The CEUS results were considered as benign tumors and confirmed by pathology. Conclusions: BBT can show ovarian cystic, mixed cystic and solid type, and solid echo in 2D ultrasound. Unilateral ovarian fibrosis with punctate calcification is an important feature of BBT in 2D ultrasound. However, for solid adnexal masses and mixed cystic and solid masses with unclear diagnosis, if CEUS shows isoenhancement or hyperenhancement, the possibility of BBT cannot be excluded.

Green Synthesis and Biosafety Assessment of MXene.

The rise of MXene-based materials with fascinating physical and chemical properties has attracted wide attention in the field of biomedicine, because it can be exploited to regulate a variety of biological processes. The biomedical applications of MXene are still in its infancy, nevertheless, the comprehensive evaluation of MXene's biosafety is desperately needed. In this review, the composition and the synthetic methods of MXene materials are first introduced from the view of biosafety. The evaluation of the interaction between MXene and cells, as well as the safety of different forms of MXene applied in vivo are then discussed. This review provides a basic understanding of MXene biosafety and may bring new inspirations to the future applications of MXene-based materials in biomedicine.

Research progress of cancer cell membrane coated nanoparticles for the diagnosis and therapy of breast cancer.

Nanoparticles (NPs) disguised in the cell membrane are a new type of biomimetic platform. Due to their ability to simulate the unique biological functions of membrane-derived cells, they have become one of the hotspots of research at home and abroad. The tumor-specific antigen antibody carried by breast cancer cell membranes can modify nanoparticles to have homologous tumor targeting. Therefore, nanoparticles wrapped in cancer cell membranes have been widely used in research on the diagnosis and treatment of breast cancer. This article reviews the current situation, prospects, advantages and limitations of nanoparticles modified by cancer cell membranes in the treatment and diagnosis of breast cancer.

Contrast-enhanced ultrasonography findings of LAMNs with peritoneal and splenic metastases: a case report and literature review.

Low-grade appendiceal mucinous neoplasms (LAMNs) are rare appendiceal tumors that are primarily diagnosed using computed tomography(CT) enhancement and magnetic resonance imaging (MRI). Herein, we report the sonographic features, especially for contrast-enhanced ultrasound (CEUS), of a 70-year-old female with an unusual LAMN metastasizing to the peritoneum and spleen. The patient had a right pelvic mass 2 days prior to presentation. Two-dimensional (2D) ultrasound revealed a mixed cystic-solid mass in the right lower abdomen and spleen parenchyma; CEUS showed heterogeneous enhancement in both areas, suspected to be a mucinous mass. CT enhancement and MRI findings revealed concurrent findings. Histopathologically, LAMN lesions were confirmed in the appendix, spleen, and peritoneum of the specimens obtained during exploratory laparoscopy. No recurrences were reported at three years postoperatively. LAMN lesions may metastasize to abdominal organs, and imaging examinations are essential for diagnosis. This study presents major ultrasonography and CEUS findings for the diagnosis of LAMNs.

3D micropattern force triggers YAP nuclear entry by transport across nuclear pores and modulates stem cells paracrine.

Biophysical cues of the cellular microenvironment tremendously influence cell behavior by mechanotransduction. However, it is still unclear how cells sense and transduce the mechanical signals from 3D geometry to regulate cell function. Here, the mechanotransduction of human mesenchymal stem cells (MSCs) triggered by 3D micropatterns and its effect on the paracrine of MSCs are systematically investigated. Our findings show that 3D micropattern force could influence the spatial reorganization of the cytoskeleton, leading to different local forces which mediate nucleus alteration such as orientation, morphology, expression of Lamin A/C and chromatin condensation. Specifically, in the triangular prism and cuboid micropatterns, the ordered F-actin fibers are distributed over and fully transmit compressive forces to the nucleus, which results in nuclear flattening and stretching of nuclear pores, thus enhancing the nuclear import of YES-associated protein (YAP). Furthermore, the activation of YAP significantly enhances the paracrine of MSCs and upregulates the secretion of angiogenic growth factors. In contrast, the fewer compressive forces on the nucleus in cylinder and cube micropatterns cause less YAP entering the nucleus. The skin repair experiment provides the first in vivo evidence that enhanced MSCs paracrine by 3D geometry significantly promotes tissue regeneration. The current study contributes to understanding the in-depth mechanisms of mechanical signals affecting cell function and provides inspiration for innovative design of biomaterials.

Field-scale assessment of direct and indirect effects of soil texture on organic matter mineralization during a dry summer.

Soil texture plays a crucial role in organic matter (OM) mineralization through both direct interactions with minerals and indirect effects on soil moisture. Separating these effects could enhance the modelling of soil organic carbon (SOC) dynamics under climate change scenarios. However, the attempts have been limited small-scale experiments. Here, we studied the effects of soil texture on added OM mineralization in loamy sand, loam and silt loam soils in nine agricultural fields in Flanders, Belgium. Soil moisture, temperature, groundwater table depth and the mineralization of 13C-labeled ryegrass were monitored in buried mesocosms for approximately three months during a dry summer. Ryegrass-C mineralization was lowest in the loamy sand (39 ± 7 %) followed by silt loam (48 ± 7 %) and loam (63 ± 5 %) soils, challenging the current clay%-based moderation of C-mineralization rates in soil models. Soil temperature was not influenced by soil texture, whereas soil moisture was indeed dependent on soil texture. It appears that capillarity sustained upward water supply from groundwater to the topsoil in loam and silt loam soils but not in loamy sand soil, although this difference in capillary rise could not fully explain the higher moisture content in loam than that in silt loam soils. Additionally, soil texture only impacted remnant added ryegrass pieces (>500 µm) but not the finer ryegrass-derived SOC (<500 µm), which might point at the important indirect control of texture on OM mineralization during prolonged summer drought. However, these effects are only manifested during drought when no other factors (e.g., groundwater depth or subsurface water flows) exert an overriding impact on the soil water balance. Overall, our findings highlight the need to properly incorporate the indirect effects of soil texture on OM mineralization into soil carbon models to accurately predict soil C stocks under future climate change scenarios.

PSMC4 promotes prostate carcinoma progression by regulating the CBX3-EGFR-PI3K-AKT-mTOR pathway.

Proteasome 26S subunit ATPase 4 (PSMC4) could regulate cancer progression. However, the function of PSMC4 in prostate carcinoma (PCa) progression requires further clarification. In the study, PSMC4 and chromobox 3 (CBX3) levels were verified by TCGA data and tissue microarrays. Cell counting kit-8, cell apoptosis, cell cycle, wound healing, transwell and xenograft tumour model assays were performed to verify biological functions of PSMC4 in PCa. RNA-seq, PCR, western blotting and co-IP assays were performed to verify the mechanism of PSMC4. Results showed that PSMC4 level was significantly increased in PCa tissues, and patients with PCa with a high PSMC4 level exhibited shorter overall survival. PSMC4 knockdown markedly inhibited cell proliferation, cell cycle and migration in vitro and in vivo, and significantly promoted cell apoptosis. Then further study revealed that CBX3 was a downstream target of PSMC4. PSMC4 knockdown markedly reduced CBX3 level, and inhibited PI3K-AKT-mTOR signalling. CBX3 overexpression markedly promoted epidermal growth factor receptor (EGFR) level. Finally, PSMC4 overexpression showed reverse effect in DU145 cells, and the effects of PSMC4 overexpression on cell proliferation, migration and clonal formation were rescued by the CBX3 knockdown, and regulated EGFR-PI3K-AKT-mTOR signalling. In conclusion, PSMC4 could regulate the PCa progression by mediating the CBX3-EGFR-PI3K-AKT-mTOR pathway. These findings provided a new target for PCa treatment.

Evolution of proton pump inhibitor prescribing from 2017 to 2021 at 14 secondary and tertiary hospitals in China：a multicentre cross-sectional study.

OBJECTIVE: The objective of this study was to describe the trend in prescribing proton pump inhibitor (PPI) use and expenditure in both secondary and tertiary hospitals in China between 2017 and 2021. DESIGN: Multicentre cross-sectional survey. SETTING: China, 14 medical centres, January 2017 to December 2021. PARTICIPANTS: 537 284 participants who were treated with PPI in 14 medical centres of China, between January 2017 and December 2021 were included. MAIN OUTCOMES AND MEASURES: The rate of PPI prescriptions, the defined daily doses (DDDs), DDDs/1000 inhabitants per day (DDDs/TID) and expenditure were analysed and plotted to demonstrate changes in prescription PPI use and expenditure. RESULTS: For both outpatient and inpatient settings, the rate of PPI prescribing decreased from 2017 to 2021. In outpatient settings, decreased slightly from 3.4% to 2.8%, however, in inpatient settings, showed a progressive decrease from 26.7% to 14.0%. The overall rate of injectable PPI prescriptions for inpatients decreased significantly from 21.2% to 7.3% between 2017 and 2021. Decreased trends in usage of oral PPI were observed (from 280 750 DDDs to 255 121 DDDs) between 2017 and 2021. However, usage of injectable PPI showed a significantly decrease from 191 451 DDDs to 68 806 DDDs from 2017 to 2021. In terms of DDDs/TID of PPI for inpatients decreased dramatically from 52.3 to 30.2 for the past 5 years. Expenditure on oral PPI decreased slightly from ¥1.98 million (Chinese currency Renminbi 'yuan') to ¥1.23 million for the past 5 years, whereas expenditure on injectable PPI showed a marked decrease from ¥2.61 million to ¥0.94 million. There was no statistical difference in both PPI use and expenditure between secondary and tertiary hospitals during the study period. CONCLUSIONS: Decreased PPI use and expenditure were observed among secondary and tertiary hospitals over the past 5 years (2017-2021).

Associations of dietary fiber intake with chronic inflammatory airway diseases and mortality in adults: a population-based study.

Objective: The objective of this study was to investigate the potential association between dietary fiber intakes and the prevalence of chronic inflammatory airway diseases (CIAD), as well as mortality in participants with CIAD. Methods: Data was collected from the National Health and Nutrition Examination Survey (NHANES) 2013-2018, with dietary fiber intakes being calculated as the average of two 24-h dietary reviews and divided into four groups. CIAD included self-reported asthma, chronic bronchitis, and chronic obstructive pulmonary disease (COPD). Through December 31, 2019, mortality was identified from the National Death Index. In cross-sectional studies, multiple logistic regressions were used to assess dietary fiber intakes associated with the prevalence of total and specific CIAD. Dose-response relationships were tested using restricted cubic spline regression. In prospective cohort studies, cumulative survival rates were calculated using the Kaplan-Meier method and compared using log-rank tests. Multiple COX regressions were used to assess dietary fiber intakes associated with mortality in participants with CIAD. Results: A total of 12,276 adults were included in this analysis. The participants had a mean age of 50.70 ± 17.4 years and was 47.2% male. The prevalence of CIAD, asthma, chronic bronchitis, and COPD were 20.1, 15.2, 6.3, and 4.2%, respectively. The median daily consumption of dietary fiber was 15.1 [IQR 10.5, 21.1] g. After adjusting for all confounding factors, linear and negative associations were observed between dietary fiber intakes and the prevalence of total CIAD (OR = 0.68 [0.58-0.80]), asthma (OR = 0.71 [0.60-0.85]), chronic bronchitis (OR = 0.57 [0.43-0.74]) and COPD (OR = 0.51 [0.34-0.74]). In addition, the fourth quartile of dietary fiber intake levels remained significantly associated with a decreased risk of all-cause mortality (HR = 0.47 [0.26-0.83]) compared to the first quartile. Conclusion: Dietary fiber intakes were found to be correlated with the prevalence of CIAD, and higher dietary fiber intakes were associated with a reduced mortality in participants with CIAD.

Establishment of a 7-gene prognostic signature based on oxidative stress genes for predicting chemotherapy resistance in pancreatic cancer.

Background: Oxidative stress is involved in regulating various biological processes in human cancers. However, the effect of oxidative stress on pancreatic adenocarcinoma (PAAD) remained unclear. Methods: Pancreatic cancer expression profiles from TCGA were downloaded. Consensus ClusterPlus helped classify molecular subtypes based on PAAD prognosis-associated oxidative stress genes. Limma package filtered differentially expressed genes (DEGs) between subtypes. A multi-gene risk model was developed using Lease absolute shrinkage and selection operator (Lasso)-Cox analysis. A nomogram was built based on risk score and distinct clinical features. Results: Consistent clustering identified 3 stable molecular subtypes (C1, C2, C3) based on oxidative stress-associated genes. Particularly, C3 had the optimal prognosis with the greatest mutation frequency, activate cell cycle pathway in an immunosuppressed status. Lasso and univariate cox regression analysis selected 7 oxidative stress phenotype-associated key genes, based on which we constructed a robust prognostic risk model independent of clinicopathological features with stable predictive performance in independent datasets. High-risk group was found to be more sensitive to small molecule chemotherapeutic drugs including Gemcitabine, Cisplatin, Erlotinib and Dasatinib. The 6 of 7 genes expressions were significantly associated with methylation. Survival prediction and prognostic model was further improved through a decision tree model by combining clinicopathological features with RiskScore. Conclusion: The risk model containing seven oxidative stress-related genes may have a greater potential to assist clinical treatment decision-making and prognosis determination.

Forms of nitrogen inputs regulate the intensity of soil acidification.

Soil acidification induced by reactive nitrogen (N) inputs can alter the structure and function of terrestrial ecosystems. Because different N-transformation processes contribute to the production and consumption of H+ , the magnitude of acidification likely depends on the relative amounts of organic N (ON) and inorganic N (IN) inputs. However, few studies have explicitly measured the effects of N composition on soil acidification. In this study, we first conducted a meta-analysis to test the effects of ON or IN inputs on soil acidification across 53 studies in grasslands. We then compared soil acidification across five different ON:IN ratios and two input rates based on long-term field N addition experiments. The meta-analysis showed that ON had weaker effects on soil acidification than IN when the N addition rate was above 20 g N m-2 year-1 . The field experiment confirmed the findings from meta-analysis: N addition with proportions of ON ≥ 20% caused less soil acidification, especially at a high input rate (30 g N m-2 year-1 ). Structural equation model analysis showed that this result was largely due to a relatively low rate of H+ production from ON as NH3 volatilization and uptake of ON and NH4 + by the dominant grass species Leymus chinensis (which are both lower net contributors to H+ production) result in less NH4 + available for nitrification (which is a higher net contributor to H+ production). These results indicate that the evaluation of soil acidification induced by N inputs should consider N forms and manipulations of relative composition of N inputs may provide an effective approach to alleviate the N-induced soil acidification.

Genome-wide DNA methylation analysis in families with multiple individuals diagnosed with schizophrenia and intellectual disability.

OBJECTIVES: Schizophrenia (SZ) and intellectual disability (ID) are both included in the continuum of neurodevelopmental disorders (NDDs). DNA methylation is known to be important in the occurrence of NDDs. The family study is conducive to eliminate the effects of relative epigenetic backgrounds, and to screen for differentially methylated positions (DMPs) and regions (DMRs) that are truly associated with NDDs. METHODS: Four monozygotic twin families were recruited, and both twin individuals suffered from NDDs (either SZ, ID, or SZ plus ID). Genome-wide methylation analysis was performed in all samples and each family. DMPs and DMRs between NDD patients and unaffected individuals were identified. Functional and pathway enrichment analyses were performed on the annotated genes. RESULTS: Two significant DMPs annotated to CYP2E1 were found in all samples. In Family One, 1476 DMPs mapped to 880 genes, and 162 DMRs overlapping with 153 unique genes were recognised. Our results suggested that the altered methylation levels of FYN, STAT3, RAC1, and NR4A2 were associated with the development of SZ and ID. Neurodevelopment and the immune system may participate in the occurrence of SZ and ID. CONCLUSIONS: Our findings suggested that DNA methylation participated in the development of NDDs by affecting neurodevelopment and the immune system.

Adversarial-based latent space alignment network for left atrial appendage segmentation in transesophageal echocardiography images.

Left atrial appendage (LAA) is a leading cause of atrial fibrillation and thrombosis in cardiovascular disease. Clinicians can rely on LAA occlusion (LAAO) to effectively prevent and treat ischaemic strokes attributed to the LAA. The correct selection of the LAAO is one of the most critical stages in the successful surgical process, which relies on the quantification of the anatomical structure of the LAA for successful intervention in LAAO. In this paper, we propose an adversarial-based latent space alignment framework for LAA segmentation in transesophageal echocardiography (TEE) images by introducing prior knowledge from the label. The proposed method consists of an LAA segmentation network, a label reconstruction network, and a latent space alignment loss. To be specific, we first employ ConvNeXt as the backbone of the segmentation and reconstruction network to enhance the feature extraction capability of the encoder. The label reconstruction network then encodes the prior shape features from the LAA labels to the latent space. The latent space alignment loss consists of the adversarial-based alignment and the contrast learning losses. It can motivate the segmentation network to learn the prior shape features of the labels, thus improving the accuracy of LAA edge segmentation. The proposed method was evaluated on a TEE dataset including 1,783 images and the experimental results showed that the proposed method outperformed other state-of-the-art LAA segmentation methods with Dice coefficient, AUC, ACC, G-mean, and Kappa of 0.831, 0.917, 0.989, 0.911, and 0.825, respectively.

The value of contrast-enhanced ultrasonography in differential diagnosis of primary testicular germ cell tumors and non-germ cell tumors over 50 years old.

Background: Unlike young and middle-aged patients, seminoma is not common in patients with primary testicular tumors over the age of 50, so it cannot follow the general ideas and norms for diagnosing and treating testicular tumors, and its characteristics need to be considered separately. Methods: The conventional ultrasonography and contrast-enhanced ultrasonography (CEUS) findings of primary testicular tumors in patients over 50 years old were retrospectively analyzed and compared with the pathological results to compare the diagnostic value of these two methods. Results: Of the 13 primary testicular tumors, 8 were primary lymphomas. Conventional ultrasound of 13 cases of testicular tumors showed hypoechoic with rich blood flow, and it was difficult to identify the type accurately. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of conventional ultrasonography in diagnosing non-germ cell tumors (lymphoma and Leydig cell tumor) were 40.0%, 33.3%, 66.7%, 14.3%, and 38.5%, respectively. CEUS findings: 7 of 8 lymphomas showed uniform hyperenhancement. 2 cases of Leydig cell tumors showed uniform high enhancement. 2 cases of seminoma and 1 case of spermatocytic tumor showed heterogeneous enhancement, with necrosis in the interior. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy rate of non-germ cell tumor diagnosis according to the non-necrotic area of CEUS were 90.0%, 100.0%, 100.0%, 75.0% and 92.3%, respectively. Compared with conventional ultrasound, the difference was statistically significant (P=0.039). Conclusions: Primary testicular tumors in patients over 50 years old are mainly lymphoma, and CEUS is significantly different between germ cell tumors and non-germ cell tumors. Compared with conventional ultrasound, CEUS can distinguish testicular germ cell tumors from non-germ cell tumors more accurately. Preoperative ultrasonography is significant for accurate diagnosis and can guide clinical treatment.

Micropatterned composite membrane guides oriented cell growth and vascularization for accelerating wound healing.

Skin defect is common in daily life, but repairing large skin defects remains a challenge. Using biomaterials to deliver biochemical or physical factors to promote skin tissue regeneration is of great significance for accelerating wound healing. Specific surface micropatterns on biomaterials could affect cell behavior and tissue regeneration. However, few studies have focused on the construction of wound healing biomaterials with surface micropatterns and their role in skin tissue regeneration. In the present study, gelatin-polycaprolactone/silk fibroin composite membranes with different micropatterns were fabricated by photolithography, including line, grid and plane micropatterns. In vitro cell experiments demonstrated that the line micropattern on the composite membrane could guide cell-oriented growth, and more importantly, promote the expression of angiogenesis-related markers and α-smooth muscle actin (α-SMA) at both gene level and protein level. In the rat full-thickness skin defect model, the composite membrane with line micropatterns increased α-SMA production and neovascularization in wounds, leading to accelerated wound contraction and healing. The current study not only suggests that composite membranes with specific micropatterns can be promising wound repair materials but also provides new insights into the importance of biomaterial surface topology for tissue regeneration.

Investigation on the mechanisms of human sperm DNA damage based on the proteomics analysis by SWATH-MS.

BACKGROUND: Spermatozoa have the task of delivering an intact paternal genome to the oocyte and supporting successful embryo development. The detection of sperm DNA fragmentation (SDF) has been emerging as a complementary test to conventional semen analysis for male infertility evaluation, but the mechanism leading to SDF and its impact on assisted reproduction remain unclear. Therefore, the study identified and analyzed the differentially expressed proteins of sperm with high and low SDF. METHODS: Semen samples from men attended the infertility clinic during June 2020 and August 2020 were analyzed, and sperm DNA fragmentation index (DFI) was detected by the sperm chromatin structure assay. Semen samples with low DFI (< 30%, control group) and high DFI (≥ 30%, experimental group) were optimized by density gradient centrifugation (DGC), and the differentially expressed proteins of obtained sperm were identified by the Sequential Window Acquisition of All Theoretical Mass Spectra Mass Spectrometry (SWATH-MS) and performed GO and KEGG analysis. RESULTS: A total of 2186 proteins were identified and 1591 proteins were quantified, of which 252 proteins were identified as differentially expressed proteins, including 124 upregulated and 128 downregulated. These differentially expressed proteins were involved in metabolic pathways, replication/recombination/repair, acrosomal vesicles, kinase regulators, fertilization, tyrosine metabolism, etc. Western blotting results showed that the expression levels of RAD23B and DFFA proteins and the levels of posttranslational ubiquitination and acetylation modifications in the experimental group were significantly higher than those in the control group, which was consistent with the results of proteomics analysis. CONCLUSIONS: Proteomic markers of sperm with high DNA fragmentation can be identified by the SWATH-MS and bioinformatic analysis, and new protein markers and posttranslational modifications related to sperm DNA damage are expected to be intensively explored. Our findings may improve our understanding of the basic molecular mechanism of sperm DNA damage.

Investigation of the mechanisms leading to human sperm DNA damage based on transcriptome analysis by RNA-seq techniques.

RESEARCH QUESTION: What are the molecular mechanisms leading to human sperm DNA damage? DESIGN: Semen samples were collected and the sperm DNA fragmentation index (DFI) was assessed. Differentially expressed RNA in spermatozoa with a high (DFI ≥30%, experimental group) or normal (DFI <30%, control group) DFI were identified by RNA-sequencing (RNA-seq) technology, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was performed. Three differentially expressed RNA related to sperm DNA damage and repair, namely PMS1, TP53BP1 and TLK2, were validated using real-time quantitative (RT-qPCR). RESULTS: A total of 19,970 expressed RNA were detected in the two groups. Compared with the control group, the expression levels of 189 RNA in the experimental group were significantly increased and those of 163 genes decreased. Gene Ontology enrichment analysis showed that these RNA were mainly concentrated in the ATPase-dependent transmembrane transport complex, extracellular exosome, somatic cell DNA recombination, protein binding, cytoplasm and regulation of localization. KEGG pathway analysis showed that these RNA were mainly related to the PI3K-Akt signalling pathway, endocytosis, p53 signalling pathway and cGMP-PKG signalling pathway. The RT-qPCR results showed that the expression levels of PMS1, TP53BP1 and TLK2 in the experimental group were significantly lower than in the control group (P = 0.01, 0.015 and 0.004, respectively), which was identical to the results of RNA sequencing. CONCLUSIONS: Differentially expressed RNA related to sperm DNA damage and repair may be identified by RNA-seq technology, which provides new insights into the understanding of sperm DNA damage and repair, and will help to discover new biomarkers related to sperm DNA damage.

Monoporous Microsphere as a Dynamically Movable Drug Carrier for Osteoporotic Bone Remodeling.

To repair systematically osteoporotic bone defects, it is important to make an effort to both diminish osteoporosis and enhance bone regeneration. Herein, a specifically monoporous microsphere (MPM) carrier encapsulating dosage-sensitive and short half-time parathyroid hormone (PTH) is constructed to tackle the issue. Compared with conventional microsphere carriers involving compact, porous, and mesoporous microspheres, the MPM is desirable to achieve precisely in situ delivery and to minimize topical accumulation. The findings show that the PTH loaded inside MPMs can be gradually released from the single hole of MPMs to improve the initial drug concentration. Also, the MPMs can self-shift with the daily movement of experimental animals to effectively reduce the topical aggregation of released drugs in vitro. In vivo evaluation further confirms that the implant of MPMs-PTH plays a dual role in stimulating the regenerative repair of the cranial defect and relieving osteoporosis in the whole body. Consequently, the current work develops a dynamically movable drug delivery system to achieve precisely in situ delivery, minimize topical accumulation, and systematically repair osteoporotic bone defects.

Genome-wide analysis of the SWEET genes in Taraxacum kok-saghyz Rodin: An insight into two latex-abundant isoforms.

Taraxacum kok-saghyz Rodin (Tk) is a promising alternative rubber-producing grass. However, low biomass and rubber-producing capability limit its commercial application. As a carbon source transporter in plants, sugar will eventually be exported transporters (SWEETs) have been reported to play pivotal roles in diverse physiological events in the context of carbon assimilate transport and utilization. Theoretically, SWEETs would participate in Tk growth, development and response to environmental cues with relation to the accumulation of rubber and biomass, both of which rely on the input of carbon assimilates. Here, we identified 22 TkSWEETs through homology searching of the Tk genomes and bioinformatics analyses. RNA-seq and qRT-PCR analysis revealed these TkSWEETs to have overlapping yet distinct tissue expression patterns. Two TkSWEET isofroms, TkSWEET1 and TkSWEET12 expressed substantially in the latex, the cytoplasm of rubber-producing laticifers as well as the rubber source. As revealed by the transient expression analysis using Tk mesophyll protoplasts, both TkSWEET1 and TkSWEET12 were located in the plasma membrane. Heterologous expressions of the two TkSWEETs in a yeast mutant revealed that only TkSWEET1 exhibited apparent sugar transport activities, with a preference for monosaccharides. Interestingly, TkSWEET12, the latex-predominant TkSWEET isoform, seemed to have evolved from a tandem duplication event that results in a cluster of six TkSWEET genes with the TkSWEET12 therein, suggesting its specialized roles in the laticifers.