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Simple Summary: This study investigates the use of Chongming goats for optimizing laparoscopic ovum pick-up (LOPU) and in vitro embryo production (IVEP). Key metrics such as the recovery rate of cumulus–oocyte complexes (COCs), the number of ovarian follicles, and the quality of collected COCs are assessed to refine estrus synchronization and superovulation techniques. Additionally, the study examines the effectiveness of LOPU using indicators like cleavage and blastocyst rates to enhance IVEP efficiency, aiming to develop a robust LOPU technology system for small ruminants. The application of laparoscopic ovum pick-up (LOPU) and in vitro production of embryos (IVEP) technologies has opened up a new path for purebred breeding and breed improvement in goats. However, due to the complexity of the procedures and multiple influencing factors, these technologies have not been widely adopted in goat production. This study explores factors affecting the efficiency of LOPU in goats by comparing the use of controlled internal drug release (CIDR) for estrus synchronization, conventional FSH versus long-acting recombinant ovine FSH (R-FSH) for superovulation, and the timing of LOPU at 48 h, 60 h, and 72 h of follicular development. The metrics evaluated included the recovery rate of cumulus–oocyte complexes (COCs), the average number of ovarian follicles, the average number of COCs, and the average number of available COCs. The results demonstrated that the efficiency of LOPU was significantly higher with two doses of R-FSH compared to the conventional FSH superovulation protocol and the control group (p < 0.05), with two doses of R-FSH providing a higher LOPU efficiency than one dose (p < 0.05). Using CIDR for estrus synchronization showed no significant difference in LOPU efficiency compared to the non-CIDR group. Similarly, the efficiency of LOPU showed no significant difference between the hormone treatments at 48 h, 60 h, and 72 h. By exploring and optimizing the factors influencing LOPU, we ultimately established a LOPU technology system for goats that meets the production needs of small ruminants. [ABSTRACT FROM AUTHOR]

The purpose of this study was to explore the potential expression regularity of POSTN in ovarian tissue. In this study, the uterus, ovarian follicles, heart, liver, spleen, lung, kidney, and muscle tissues of Merino sheep (10) were collected and detected by Western blot, real-time quantitative polymerase chain reaction, immunohistofluorescence, and fluorescence staining, respectively. The expression of POSTN in various organizations was analyzed. The results showed that the POSTN in Merino sheep had close homology with Bos mutus and Bos taurus. The expression of POSTN was detected in the uterus, follicle, heart, liver, spleen, lung, kidney, and muscle tissue, among which the expression of POSTN was highest in ovarian tissue. In addition, the expression of POSTN gradually increased with the increase of follicle diameter, among which POSTN was highly expressed in the granulosa cells (GCs) of follicles. Meanwhile, POSTN were distributed throughout the nucleus and cytoplasm of GCs, suggesting that POSTN may be involved in the regulation of follicle development. [ABSTRACT FROM AUTHOR]

Thrombospondin (TSP1) plays an important role as an antiangiogenic factor in the reproductive system of female mammals. However, its expression and function in sheep are still unclear. In the present research, the Altay sheep (a native Chinese breed) was used to analyze the expression of TSP1 in the ovary and its potential function in granulosa cells. TSP1 was widely expressed in most tissues, as shown by qPCR. In the ovary, TSP1 mRNA expression decreased during follicular to luteal growth. The TSP1 protein was expressed in a wide variety of follicles of different diameters and localized to the cytoplasm and nucleus of granulosa cells. In in vitro studies, follicle-stimulating hormone (FSH) significantly inhibited the expression of TSP1 in sheep granulosa cells. Functionally, FSH- and TSP1-specific siRNAs can promote the proliferation of sheep granulosa cells. In contrast, TSP1 mimetic peptide, ABT510, offsets the proliferation of sheep granulosa cells. Different signaling pathway inhibitors all promoted FSH-inhibited TSP1 expression, but each inhibitor had different effects on TSP1. Among them, the PI3K and ERK pathway inhibitors significantly promoted TSP1 expression and inhibited the proliferation of sheep granulosa cells. [ABSTRACT FROM AUTHOR]

A novel Bi2WO6 and BiFeO3 heterojunction composites with enhanced visible-light response ability have been successfully synthesized via a facile solvothermal route, which possessed high heterogeneous photo-Fenton for degrading rhodamine B dye. The results of characterizations suggested that the composites of Bi2WO6 and BiFeO3 not only have been synthesized, but also the heterojunction structure have been formed. It was found that the stepped construction of Bi2WO6/BiFeO3 heterojunction can minify the band gap to promote the separation and migration of the photogenerated electron–hole pairs, thus Bi2WO6/BiFeO3 exhibited better visible-light response and photocatalytic activity than that of pure Bi2WO6, degrading Rhodamine B (RhB) 98.07% within 75 min. BiFeO3 did not exhibited the visible photocatalytic activity as same as Bi2WO6, however, it was illustrated that the heterogeneous photo-Fenton reaction can be facilitated through coupling BiFeO3. Moreover, the enhanced photocatalytic activity of Bi2WO6/BiFeO3 could further promote the Fe2+/Fe3+ cycling to form an excellent photo-Fenton system. Hence, photo-Fenton system of Bi2WO6/BiFeO3 could generate more free radicals to more efficiently degrade RhB, achieving nearly 100% after 20 min of treatment. [ABSTRACT FROM AUTHOR]

Research is presented demonstrating bis-(oligonucleotide) conjugates which can adopt stable hairpin structures of B-form DNA. Factors influencing hairpin stability include linker length and flexibility and chromophore pi-stacking ability.

Purpose: Atractylodes macrocephala Koidz is a widely used classical traditional Chinese herbal medicine, that has shown remarkable efficacy in cancers. Colorectal cancer (CRC) is the most common malignant tumor globally. Interferon (IFN)-γ, a prominent cytokine involved in anti-tumor immunity that has cytostatic, pro-apoptotic, and immune-stimulatory properties for the detection and removal of transformed cells. Atractylenolides-II (AT-II) belongs to the lactone compound that is derived from Atractylodes macrocephala Koidz with anti-cancer activity. However, whether AT-II combined with IFN-γ modulates CRC progression and the underlying mechanisms remain unclear. The present study aimed to elucidate the efficacy and pharmaceutical mechanism of action of AT-II combined with IFN-γ synergistically against CRC by regulating the NF-kB p65/PD-L1 signaling pathway. Methods: HT29 and HCT15 cells were treated with AT-II and IFN-γ alone or in combination and cell viability, migration, and invasion were then analyzed using Cell Counting Kit-8 (CCK-8) and Transwell assays, respectively. Furthermore, the underlying mechanism was investigated through western blot assay. The role of AT-II combined with IFN-γ on tumor growth and lung metastases was estimated in vivo . Finally, the population of lymphocytes in tumor tissues of lung metastatic C57BL/6 mice and the plasma cytokine levels were confirmed by flow cytometry and enzyme-linked immunosorbent assay (ELISA). Results: AT-II or the combination IFN-γ significantly inhibited the growth and migration abilities of CRC cells in vitro and in vivo . The biological mechanisms behind the beneficial effects of AT-II combined with IFN-γ were also measured and inhibition of p38 MAPK, FAK, Wnt/β-catenin, Smad, and NF-kB p65/PD-L1 pathways was observed. Moreover, AT-II combined with IFN-γ significantly inhibited HCT15 xenograft tumor growth and lung metastases in C57BL/6 mice, which was accompanied by lymphocyte infiltration into the tumor tissues and inflammatory response inactivation. Conclusions: The results showed that the AT-II in combination with IFN-γ could be used as a potential strategy for tumor immunotherapy in CRC. More importantly, the mechanism by which AT-II suppressed CRC progressions was by inhibiting the NF-kB p65/PD-L1 signal pathway., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Coat color is an important characteristic and economic trait in domestic sheep. Aiming at alteration of Chinese merino sheep coat color by genome manipulation, we disrupted sheep agouti signaling protein gene by CRISPR/Cas9. A total of seven indels were identified in 5 of 6 born lambs. Each targeted lamb happened at least two kinds of modifications, and targeted lambs with multiple modifications displayed variety of coat color patterns. Three lambs with 4 bp deletion showed badgerface with black body coat color in two lambs, and brown coat color with light ventral pigmentation in another one. The black-white spotted color was observed in two lambs with 2 bp deletion. Further analysis unraveled that modifications happened in one or more than two copies of ASIP gene, and moreover, the additional spontaneous mutations of D 9 and/or D 5 preceding the targeting modification could also involve the formation of coat color patterns. Taken together, the entanglement of ASIP modifications by CRISPR/Cas9, spontaneous D 9 /D 5 mutations, and ASIP gene duplications contributed to the variety of coat color patterns in targeted lambs.

Objective: Superstimulatory treatment of one-month-old lambs can achieve synchronous development of numerous growing follicles. However, these growing follicles cannot complete maturation and ovulation. Oocyte maturation and competence are acquired during follicular development, in which granulosa cells play an essential role., Methods: In this study, we applied RNA sequencing to analyze and compare gene expression between prepubertal and adult superstimulated follicle granulosa cells in sheep., Results: There were more than 300 genes that significantly differed in expression. Among these differently expressed genes, many extracellular matrix genes (EGF containing Fibulin Like Extracellular Matrix Protein 1, pentraxin 3, adrenomedullin, and osteopontin) were significantly down-regulated in the superstimulated follicles. Ingenuity pathway and gene ontology analyses revealed that processes of axonal guidance, cell proliferation and DNA replication were expressed at higher levels in the prepubertal follicles. Epidermal growth factor, T-Box protein 2 and beta-estradiol upstream regulator were predicted to be active in prepubertal follicles. By comparison, tumor protein P53 and let-7 were most active in adult follicles., Conclusion: These results may contribute to a better understanding of the mechanisms governing the development of granulosa cells in the growing follicle in prepubertal sheep.