Analysis of use of different rFSHs during IVF/ICSI-assisted conception in elderly population and effect of double trigger on clinical outcomes.

OBJECTIVE: To compare the number of oocytes retrieved and clinical outcomes of ovulation induction in an older population treated with in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) (IVF/ICSI) using different rFSH options and the effectiveness of antagonist treatment to induce ovulation using gonadotropin-releasing hormone agonists (GnRH-a) in combination with an human chorionic gonadotropin (HCG) trigger. METHODS: A total of 132 fresh cycles were selected for this study, which were treated with IVF/ICSI in our hospital from March 2022 to December 2022. Observations were made according to different subgroups and the effects of different triggering methods on the number of oocytes obtained, embryo quality, and clinical outcomes. RESULTS: The initial gonadotropin (Gn) dose, the number of oocytes, and the number of MII oocytes were higher in group A than in group B (p < .05), and the clinical pregnancy rate was 29.41% in group A. Group B had a clinical pregnancy rate of 27.5%. The double-trigger group was superior to the HCG-trigger group in terms of the number of 2PN, the number of viable embryos, and the number of high-quality embryos (p < .05). The use of a double-trigger regimen (OR = 0.667, 95%CI (0.375, 1.706), p = .024) was a protective factor for the clinical pregnancy rate, whereas AFC (OR = 0.925, 95%CI (0.867, 0.986), p = .017) was an independent factor for the clinical pregnancy rate. CONCLUSIONS: The use of a dual-trigger regimen of GnRH-a in combination with HCG using an appropriate antagonist improves pregnancy outcomes in fresh embryo transfer cycles in older patients.

Surface charge-induced electrospray for high-throughput analysis of complex samples and electrochemical reaction intermediates using mass spectrometry.

Electrospray-related ion sources are promising for direct mass spectrometric analysis of complex samples, but current protocols suffer from complicated components and low analytical sensitivity. Here, we propose a surface charge-induced electrospray ionization (SCIESI) inspired by flashover on an insulator surface under high voltage. This protocol not only effectively avoids contact between the sample solution and metal electrode, but also allows completion of the entire analytical process in less than 40 seconds and limits of detection in the pictogram per milliliter range. SCIESI coupled to mass spectrometry can also be used to monitor electro-chemical processes, and a number of oxidation and reduction reactions have been studied, demonstrating that it is a powerful tool for understanding electrochemical reaction mechanisms.

Influence of internal angle and shape of the lining on residual stress of Class II molar restorations.

Polymerization shrinkage is a major side effect of resin composite materials that affects the success and longevity of caries restorations. This study was to analyze the effect of the internal angle and shape of the lining on the shrinkage residual stress of dental restorations in Class II mesio-occlusal-distal (MOD) cavities through finite element analysis (FEA). A 3D reconstructed model of a human mandibular first molar was created from micro-CT images, and then the tooth was prepared as a Class II MOD cavity. 3D models of four regular internal corner shapes of Class II MOD cavities with different internal angles of lining and one 90° filleted corner model were created. The thermal expansion technique was applied to approximate the shrinkage impact of composite resin polymerization in the FE software ABAQUS. Von Mises stress was taken as a metric. The results showed that the level of residual stresses in the Class II MOD cavities was greatly dependent on the internal angle of the lining. The maximum von Mises stress in tooth tissue decreased as the internal angle of the lining increased. The internal shapes of the lining had no obvious effect on the stress, and the filleted corner model had less stress. This lining strategy is appropriate for Class II MOD cavities with serious loss of tooth tissue.

Influence of ZnO morphology on the capability of portable paper-based electrospray ionization mass spectrometry to determine therapeutic drugs in complex matrices.

Adsorbents play a significant role in enhancing the analytical sensitivity of target analytes in complex samples by mitigating matrix effects. In our recent report, ZnO stood out among various adsorbents to determine target therapeutic drugs in complex biological matrices when applied for portable paper-based electrospray ionization mass spectrometry (PPESI-MS). However, the influence of the morphology of ZnO on the performance of PPESI-MS is elusive. Herein, different morphologies of ZnO particles were prepared via co-precipitation or ultrasonic methods, and their capability to determine different therapeutic drugs in serum were systemically investigated. The results demonstrated that flower-shaped ZnO gave a superior capacity, and its analysis sensitivity was 2.9-12.8-fold higher than those achieved with other ZnO morphologies. Further characterization revealed that the unique performance of flower-shaped ZnO was closely associated with its favorable desorption behavior to drugs, small spray plume, and few spray emitters at the tip of coated paper substrate. To illustrate the potential of flower-shaped ZnO, its coated paper was used as a substrate for the determination of various drugs in complex matrices such as serum, and a limit of detection as low as 2 pg mL-1 was achieved. The corresponding recoveries ranged from 93.2% to 107.2%. The developed protocol is promising in high-sensitivity analysis of target drugs in complex sample matrices.

Ultralow background one-pot detection of Lead(II) using a non-enzymatic double-cycle system mediated by a hairpin-involved DNAzyme.

A double-cycle system has been developed for specifically detecting trace amounts of Pb2+ by significantly decreasing the background signal. The detection involves two types of RNA cleavage reactions: one using a Pb2+-specific GR5 DNAzyme (PbDz) and the other utilizing a newly constructed 10-23 DNAzyme with two hairpins embedded in its catalytic center (hpDz). The ring-structured hpDz (c-hpDz) exhibits significantly lower activity compared to the circular 10-23 DNAzyme without hairpin structures, which plays a crucial role in reducing the background signal. When Pb2+ is present, PbDz cleaves c-hpDz to its active form, which then disconnects the molecular beacon to emit the fluorescent signal. The method allows for rapid and sensitive Pb2+ detection within 40 min for 10 fM of Pb2+ and even as short as 10 min for 100 nM of Pb2+. Additionally, visual detection is possible through the non-crosslinking assembly of Au nanoparticles. The entire process can be performed in one pot and even one step, making it highly versatile and suitable for a wide range of applications, including food safety testing and environmental monitoring.

Programming Affinity for Precise Tumor Recognition with Allosteric Nanosensing-Circles.

Although many smart probes for precise tumor recognition have been reported, the challenge of "on-target, off-tumor" remains. Therefore, we herein report the fabrication of a series of allosterically tunable DNA nanosensing-circles (NSCs). The recognition affinity of NSCs is programmed through sensitivity to tumor microenvironment (TME) hallmarks such as small molecules, acidity, or oncoproteins. Because of their special programming conditions and active targeting capabilities, NSCs can overcome the obstacles noted above, thus achieving precise tumor recognition. Results from in vitro analysis demonstrated that NSCs obtain their recognition ability through allosteric regulation after sensing TME hallmarks. Furthermore, in vivo imaging indicated that NSCs enable precise tumor imaging. These results demonstrate that our NSCs will be promising tools for precise tumor imaging and therapy.

On-Demand Portable Paper-Based Electrospray Ionization Mass Spectrometry for High-Sensitivity Analysis of Complex Samples.

Paper spray ionization has been demonstrated to be the most promising substrate-based source, but this technique suffers from the low desorption efficiency of target compounds and poor portability. In the current study, we describe a portable paper-based electrospray ionization (PPESI) in which a piece of triangle paper and adsorbent are packed sequentially into a modified disposable micropipette tip. This source not only captures the feature of paper spray and adsorbent for highly efficient suppression of sample matrixes for target compound analysis but also takes advantage of a micropipette tip to prevent spray solvent from rapid evaporation. The performance of developed PPESI depends on the type and amount of packed adsorbent, paper substrate, and spray solvent and applied voltage. Moreover, by contrast to other related sources, the analytical sensitivity and the spray duration of PPESI in tandem with MS have been improved by factors of 2.8-32.3 and 2.0-13.3, respectively. Based on its high accuracy (>96%) and precision (less than 3% relative standard deviation), the PPESI coupled to a mass spectrometer has been used to determine diverse therapeutic drugs and pesticides in complex biological (e.g., whole blood, serum, and urine) and food (e.g., milk and orange juice) matrixes, and the limits of detection and quantification were 2-4 pg mL-1 and 7-13 pg mL-1, respectively. Taking the portability, high sensitivity, and repeatability, the technique may be a promising alternative for complex sample analysis.

Repeated controlled ovarian stimulation-induced ovarian and uterine damage in mice through the PI3K/AKT signaling pathway.

The effects of repeated controlled ovarian stimulation (COS) on the female reproductive system are still controversial. This study investigated the effects of repeated COS on the ovaries and uterus of mice and its possible mechanism. Female ICR (Institute of Cancer Research) mice were subjected to the COS using pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) for 1, 3, 5, and 7 cycles. Serum hormone levels, reactive oxidative stress (ROS), 8-hydroxy-2'-deoxyguanosine (8-OHdG), total antioxidant capacity (T-AOC), and superoxide dismutase (SOD) in the mouse ovary and uterus were analyzed by ELISA. The morphology of the ovary and endometrium, ovarian apoptosis, and expressions of the vascular endothelial growth factor (VEGF), leukemia inhibitory factor (LIF), PI3K, AKT, Bax, and Bcl-2 in the ovarian and uterine tissues were tested by hematoxylin-eosin (HE) staining, immunohistochemistry, and western blot. The results showed that repeated COS significantly decreased the hormone level (estradiol, progesterone and anti-Müllerian hormone), high-quality of the MII oocyte ratio, oocyte and embryo number, antioxidant capacity (T-AOC, SOD activity), and the protein level of Bcl-2, LIF, and VEGF, but increased the oxidative damage (ROS, 8-OHdG content), embryo fragment ratio, and expression of pro-apoptotic protein Bax. In addition, the expressions of p-PI3K and p-AKT also decreased with the increase of COS cycle. In conclusion, repeated COS causes ovarian and uterus damage possibly through the PI3K/AKT signaling pathway, and this finding may provide some experimental basis for guiding clinical treatment.

Improvement in the performance of focusing plasma desorption ionization by altering its counter electrode.

RATIONALE: Plasma-based ionization sources play a vital role in rapidly analyzing diverse compounds without extensive sample pretreatment. In contrast to other sources, DC voltage-based ionizations are more advantageous due to their high analytical sensitivity and good tandem with commercially available mass spectrometers without extra power supplies. However, their performance is at the risk of high current DC voltage and helium flow rate, which poses significant challenges to practical operation and increased expense. METHODS: In this work, we propose a novel focusing plasma desorption ionization (FPDI) in which a visible plasma beam is favorably generated between a conducting wire in a polymeric tube and a counter electrode composed of metal mesh and filter paper drilled with holes. A systematic investigation has been conducted on the influences of the geometry of drilled holes in filter paper, applied DC voltage, helium flow rate, and filter paper size. The optimized system is used to analyze various pesticides in fluid foodstuffs. RESULTS: Compared to metal mesh and conducting paper as the counter electrode for FPDI-MS, combining metal mesh and filter paper drilled holes improved the analysis sensitivity by a factor of more than five. By applying the developed protocol for determining pesticides in complex matrixes such as orange juice and milk, a limit of detection as low as 1.3-3.0 ng mL-1 could be achieved. CONCLUSIONS: A novel FPDI-MS technique has been developed by combining metal mesh and filter paper drilled with holes as the counter electrode and sample carrier. The corresponding improvement in analysis sensitivity facilitates the future expansion of FPDI-MS applications into different pesticides and other compounds in complex matrixes.

HDAC19 recruits ERF4 to the MYB5a promoter and diminishes anthocyanin accumulation during grape ripening.

DNA acetylation alters the expression of responsive genes during plant development. In grapes (Vitis vinifera), however, little is known about this regulatory mechanism. In the present study, 'Kyoho' grapes treated with trichostatin A (TSA, a deacetylase inhibitor) were used for transcriptome sequencing and quantitative proteomics analysis. We observed that acetylation was associated with anthocyanin accumulation and gene expression. Acetylation positively regulated phenylalanine metabolism and flavonoid biosynthesis pathways. Using omics analysis, we detected an increase in the levels of the AP2/EREBP transcription factor family after TSA treatment, indicating its association with acetylation-deacetylation dynamics in grapes. Furthermore, ethylene response factor 4 (ERF4) physically interacted with VvHDAC19, a histone deacetylase, which synergistically reduced the expression of target genes involved in anthocyanin biosynthesis owing to the binding of VvERF4 to the GCC-box cis-regulatory element in the VvMYB5a promoter. VvHDAC19 and VvERF4 also controlled anthocyanin biosynthesis and accumulation by regulating acetylation levels of histones H3 and H4. Therefore, alterations in histone modification can significantly regulate the expression of genes involved in anthocyanin biosynthesis and affect grape ripening.

Label-free based quantitative proteomics analysis to explore the molecular mechanism of gynecological cold coagulation and blood stasis syndrome.

Cold coagulation and blood stasis (CCBS) syndrome is one of the common traditional Chinese medicine (TCM) syndromes of gynecological diseases. However, the molecular mechanism of CCBS syndrome is still unclear. Thus, there is a need to reveal the occurrence and regulation mechanism of CCBS syndrome, in order to provide a theoretical basis for the treatment of CCBS syndrome in gynecological diseases. The plasma proteins in primary dysmenorrhea (PD) patients with CCBS syndrome, endometriosis (EMS) patients with CCBS syndrome, and healthy women were screened using Label-free quantitative proteomics. Based on the TCM theory of "same TCM syndrome in different diseases," the differentially expressed proteins (DEPs) identified in each group were subjected to intersection mapping to obtain common DEPs in CCBS syndrome. The DEPs of gynecological CCBS syndrome in the intersection part were again cross-mapped with the DEPs of gynecological CCBS syndrome obtained by the research group according to the TCM theory of "different TCM syndromes in same disease" theory in the early stage, so as to obtain the DEPs of gynecological CCBS syndrome that were shared by the two parts. The common DEPs were subjected to bioinformatics analysis, and were verified by enzyme-linked immunosorbent assay (ELISA). A total of 67 common DEPs were identified in CCBS syndrome, of which 33 DEPs were upregulated and 34 DEPs were downregulated. The functional classification of DEPs involved in metabolic process, energy production and conversion, immune system process, antioxidant activity, response to stimulus, and biological adhesion. The subcellular location mainly located in the cytoplasm, nucleus, and extracellular. Gene ontology (GO) enrichment analysis showed that the upregulated DEPs mainly concentrated in lipid transport, cell migration, and inflammatory reaction, and the downregulated DEPs mostly related to cell junction, metabolism, and energy response. Protein domain enrichment analysis and clustering analysis revealed that the DEPs mainly related to cell proliferation and differentiation, cell morphology, metabolism, and immunity. The Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis clustering analysis showed that the upregulated DEPs were involved in inflammation and oxidative damage, while the downregulated DEPs were involved in inflammation, cell adhesion, cell apoptosis, and metabolism. The results of ELISA showed significantly increased levels of Cell surface glycoprotein MUC18 (MCAM) and Apolipoprotein C1 (APOC1), and significantly decreased levels of Vasodilator-stimulated phosphoprotein (VASP), Fatty acid-binding protein 5 (FABP5), and Vinculin (VCL) in patients with CCBS syndrome compared with healthy women. We speculated that cold evil may affect the immune process, inflammatory response, metabolic process, energy production and conversion, oxidative damage, endothelial cell dysfunction, and other differential proteins expression to cause CCBS syndrome in gynecological diseases.

Clinical efficacy of Wenjing decoction in the treatment of ovulatory disorder infertility: A systematic review and meta-analysis.

OBJECTIVES: Wenjing decoction (WJD) was widely used in the treatment for ovulatory disorder infertility (ODI) in China, while its efficacy was not clearly known. In this study, we evaluated the clinical efficacy of WJD by meta-analysis. METHODS: Eight electronic databases including Cochrane Library, PubMed, Embase, Web of Science, China National Knowledge Infrastructure, WanFang Data, VIP Database, and China Biology Medicine were searched for randomized controlled trials (RCTs) published from the inception of each database to July 1, 2021, of which the interventions involve WJD and clomiphene. Outcomes included clinical efficacy rate, pregnancy rate, ovulation rate, dominant follicle diameter, endometrial thickness, estradiol, follicle-stimulating hormone, and luteinizing hormone. Meta-analysis and risk of bias were performed by RevMan 5.3 software. RESULTS: Eleven RCTs including 915 patients, of which 476 in the intervention group and 439 in the control group. Meta-analysis showed that WJD was better than clomiphene for patients with ODI in terms of clinical effective rate (odds ratio [OR] = 1.22, 95% confidence interval [CI]: 1.08-1.34), pregnancy rate (OR = 1.54, 95% CI: 1.15-2.07), ovulation rate (OR = 1.34, 95% CI: 1.07-1.67), endometrial thickness (mean difference [MD] = 1.50, 95% CI: 0.90-2.10), and dominant follicle diameter (MD = 1.85, 95% CI: 0.68-3.02). The estradiol level (MD = 91.0, 95% CI: 80.3-101.88) in patients taking WJD was significantly higher than those taking clomiphene, while the follicle-stimulating hormone level (MD = -0.93, 95% CI: -1.13 to -0.72) and the luteinizing hormone level (MD = -4.41, 95% CI: -4.80 to -4.03) in patients taking WJD was significantly lower than those taking clomiphene. Our results also indicated that WJD combined with clomiphene was better than clomiphene alone for patients with ODI in terms of pregnancy rate (OR = 1.79, 95% CI: 1.37-2.35). CONCLUSIONS: WJD may be effective in the treatment of patients with ODI. Due to the quality and quantity of literature, RCT with large sample size and high quality need to be performed to verify our conclusion.

Clinical application of oral Chinese Patent Medicines in ophthalmology: a scoping review protocol.

INTRODUCTION: The prevalence of eye diseases has been increasing worldwide. In China, in addition to conventional medicine, Traditional Chinese Medicine (TCM) plays an important role in maintaining people's vision health. Although less flexible and targeted than TCM decoction, Chinese Patent Medicines (CPMs) are stable and well used. In recent years, CPMs have been increasingly used in ophthalmology clinics by TCM practitioners and by Western doctors in general hospitals. However, comprehensive evidence for using CPMs in ophthalmology is lacking. AIM: We will apply the methodology of scoping review to systematically search and sort out the available evidence on oral CPMs for the treatment of eye diseases, identify the distribution of evidence in this field and provide a basis for clinical practice and medical decisions. METHODS: The scoping review will be implemented in the following seven steps: (1) defining the research question; (2) searching National Essential Medicines List (2018 edition), National Basic Medical Insurance, Work Injury Insurance and Maternity Insurance Drug Catalog (2020 edition) and Chinese Pharmacopoeia (2020 edition) for oral CPMs for the treatment of eye diseases; (3) searching Embase, Web of Science, PubMed, Cochrane Library, Chongqing VIP Chinese Scientific Journals Database, Chinese National Knowledge Infrastructure, Chinese Biomedical Literature Database and Wanfang Database for relevant literature published from inception to December 2021; (4) developing eligibility criteria; (5) screening the studies based on inclusion criteria; (6) extracting relevant data and lastly, (7) collating, summarising and reporting the results. ETHICS AND DISSEMINATION: Since the scoping review aims at collecting data from publicly available publications, this study does not require ethical approval. The results will be published in a peer-reviewed journal and presented at scientific conferences.

Construction and evaluation of prognostic models for esophageal cancer patients with distant and non-distant metastases: providing a reference process for clinical diagnosis and treatment.

BACKGROUND: Although the current treatment for esophageal cancer has great technological progress, the 5-year survival rate of patients is not optimistic. About 70% of patients with esophageal cancer are at an advanced stage at first diagnosis. These patients are prone to distant metastasis, and the prognosis is poor. Therefore, understanding the risk factors for distant metastasis in patients with esophageal cancer, combined with the prognosis of the patient, can aid in choosing the optimal diagnosis and treatment plan. Ultimately, it will improve the patient's survival time and quality of life. This research aims to construct a model for the risk assessment of distant metastasis in patients with esophageal cancer and prognostic models for patients with distant and non-distant metastases. METHODS: The Surveillance Epidemiology and End Results (SEER) database was used to select patients with esophageal cancer from 2010 to 2015. The optimal cutoff point was selected for the age and tumor size variables using X-tile. The nomogram was constructed using R software (The R Foundation for Statistical Computing). RESULTS: Gender, grade, T stage, N stage, and tumor size were independent risk factors associated with distant metastasis in patients with esophageal cancer. The concordance index (C-index) of the nomogram prediction model for whether the patient will have distant metastasis was 0.609. Age, grade, T stage, N stage, and tumor size were independent risk factors affecting the prognosis without distant metastasis. The C-index of the nomogram prediction model for patients with distant metastases was 0.590. Age and T stage were independent risk factors affecting the prognosis of patients with distant metastases. The C-index of the nomogram prediction model was 0.543. The combination of radiotherapy, chemotherapy, and primary surgery yielded the best overall survival for both patients with distant metastases and patients with non-distant metastases. CONCLUSIONS: A comprehensive assessment of the risk of distant metastasis in patients with esophageal cancer, combined with prognosis prediction, is necessary to provide patients with a reasonable treatment plan.

[Effects of ant colonization on spatiotemporal variation of organic carbon mineralization in Xishuangbanna tropical forest soils]. / 蚂蚁筑巢对西双版纳热带森林土壤碳矿化动态的影响.

Ants as ecosystem engineers can increase the input of soil organic matter, change soil physicochemical properties, and stimulate microbial activities through their colonization, thus affecting the spatiotemporal dynamics of soil organic carbon mineralization. We explored the spatiotemporal characteristics of carbon mineralization rates in ant nests and the adjacent soils in Syzygium oblatum community of Xishuangbanna, Yunnan. We analyzed the association of the variation in carbon mineralization rates with soil physicochemical properties. We found that ant colonization had a significant effect on soil organic carbon mineralization. The mean carbon mineralization rate was 19.2% higher in nest soils than that in the surrounding soils. The monthly carbon mineralization rate in nest soils and the reference soils was ranked as June ＞ September ＞ March ＞ December. The highest increase of carbon mineralization rate in ant nests was observed in 10-15 cm soil layer, while that in the reference soils was in 0-5 cm soil depth. Ant colonization had a significant effect on soil physicochemical properties. Compared with reference soils, soil temperature, soil water, soil organic carbon,soil microbial carbon, total nitrogen, hydrolytic nitrogen, nitrate, and ammonium increased by 7.6%, 5.4%, 9.9%, 14.8%, 13.4%, 9.9%, 24.1%, 6.6% and 19.4%, respectively. In contrast, soil bulk density and soil pH were decreased by 1.4% and 2.5%, respectively. Results from correlation coefficients and principal component analysis (PCA) showed that soil organic carbon and soil microbial carbon were the key factors controlling the mineralization of soil organic carbon, followed by total nitrogen, hydrolyzed nitrogen, ammonium, nitrate, temperature, and soil moisture. We conclude that ant colonization mainly alter the substrate components (i.e., soil organic carbon and microbial biomass carbon) of soil organic carbon mineralization and thus affect its spatio-temporal dynamics in Xishuangbanna tropical forests.

Construction of Nomogram prediction model for prognosis of gastric neuroendocrine neoplasms / 中国肿瘤生物治疗杂志

@# Objection: To analyze the factors affecting the prognosis of patients with gastric neuroendocrine neoplasms (G-NENs) by using the surveillance of National Cancer Institute (NCI) of America, Epidemiology and End Results (SEER) database, and to construct a prognostic Nomogram model for individualized prediction of prognosis in patients with G-NENs. Methods: The clinical data of 2720 G-NENs patients with complete follow-up data from 2010 to 2015 in the SEER database were collected. The prognostic Nomogram model was constructed based on independent risk factors determined by survival analysis. The consistency index (C-index) and calibration curve were used to evaluate its accuracy.Area under the curve (AUC) was used to compare the evaluation value between the Nomogram and the 7th edition of AJCC TNM staging. Results: The 1-, 3-, and 5-year survival rates of 2,720 patients with G-NENs were 88.14%, 79.09%, and 71.86%, respectively. Multivariate COX regression analysis showed that gender, age, marital status, other associated tumors, histological type, tumor grade, T stage, M stage, and surgery were independent risk factors affecting survival time of GNENs patients. The C-index of newly constructed Nomogram prediction model was 0.816, which was significantly higher than 0.702 of the 7thAJCC TNM staging (P<0.001), and the 1-, 3- and 5-year calibration curves showed a good agreement between predicted survival and actual survival. The AUC for 1-, 3- and 5-year survival by Nomogram prognostic model was 0.800, 0.811, and 0.820, which was higher than 0.650, 0.688 and 0.698 of the 7th AJCC TNM staging, and the differences were statistically significant (Z= 6.600, 8.085, 9.632, all P<0.0001). Conclusion: The Nomogram prediction model drawn in this study has a high prognostic value and can individually predict the survival rate of G-NENs patients, which is helpful for clinical treatment decision-making and clinical research options.

Expression and effects of TSP50 in mouse embryo and cardiac myocyte development.

TSP50, a testis-specific gene encoding a serine protease-like protein, was specifically expressed in the spermatocytes of testes but abnormally activated and expressed in many different kinds of cancers. Here, we aimed to analyze the expression of TSP50 in mouse embryo and its function in early embryonic development. Firstly, the distribution of TSP50 in oocytes and embryonic development was characterized by immunofluorescence, RT-PCR and western blotting, and the results showed that TSP50 was detected at all studied stages with a dynamic expression pattern. When overexpressed TSP50 in zygotes by microinjection, the zygotes development was highly accelerated. On the contrary, knocking down TSP50 expression by RNA interference greatly retarded the zygote development. Furthermore, TSP50 expression at embryonic day 6.5 (E6.5), day 8.5 (E8.5) and day 10.5 (E10.5) were increasingly enhanced, However, the expression of TSP50 decreased gradually in the development and differentiation of cardiac myocyte from E12.5 to postnatal (P0). Additionally, we found that TSP50 expression was decreased during cardiac myocyte differentiation of P19 cells. Overexpression of TSP50 could decrease the expression of GATA-4, and knockdown of TSP50 markedly increase the expression of GATA-4. Taken together, our data indicate that TSP50 may play an important role during the process of mouse embryonic development as well as myocardial cell differentiation.