Bta-miR-301a targets ACVR1 to influence cleavage time and blastocyst formation rate of early embryos in cattle.

Accumulating evidence indicates that paternally-derived miRNAs play a crucial role in the development of early embryos and are regarded as the key factor in the successful development of somatic cell cloned embryos. In our previous study, bta-miR-301a was found to be highly expressed in bovine sperm, and was delivered into oocytes during fertilization. In this study, bioinformatics, dual luciferase reporter assays, rescue experiments and gain- and loss-of-function experiments indicated that ACVR1 is the target gene of bta-miR-301a in early bovine embryos. By microinjecting bta-miR-301a mimic into embryos of parthenogenetic or somatic cell nuclear transfer, we observed that bta-miR-301a prolonged the first cleavage time of the embryos and increased the blastocyst formation rate. Thus, this study provides preliminary evidence that bta-miR-301a influences remodeling of the microfilament skeleton, prolongs the first cleavage time, and improves the developmental competence of embryos by negatively regulating ACVR1 translation.

Detection of the dominant pathogens in diarrheal calves of Ningxia, China in 2021-2022.

Introduction: Calf diarrhea is a complex disease that has long been an unsolved problem in the cattle industry. Ningxia is at the forefront of China in the scale of cattle breeding, and calf diarrhea gravely restricts the development of Ningxia's cattle industry. Methods: From July 2021 to May 2022, we collected diarrhea stool samples from calves aged 1-103 days from 23 farms in five cities in Ningxia, and performed PCR using specific primers for 15 major reported pathogens of calf diarrhea, including bacteria, viruses, and parasites. The effect of different seasons on the occurrence of diarrhea in calves was explored, the respective epidemic pathogens in different seasons were screened, and more detailed epidemiological investigations were carried out in Yinchuan and Wuzhong. In addition, we analyzed the relationship between different ages, river distributions and pathogen prevalence. Results: Eventually, 10 pathogens were detected, of which 9 pathogens were pathogenic and 1 pathogen was non-pathogenic. The pathogens with the highest detection rate were Cryptosporidium (50.46%), Bovine rotavirus (BRV) (23.18%), Escherichia coli (E. coli) K99 (20.00%), and Bovine coronavirus (BCoV) (11.82%). The remaining pathogens such as Coccidia (6.90%), Bovine Astrovirus (BoAstV) (5.46%), Bovine Torovirus (BToV) (4.09%), and Bovine Kobuvirus (BKoV) (3.18%) primarily existed in the form of mixed infection. Discussion: The analysis showed that different cities in Ningxia have different pathogens responsible for diarrhea, with Cryptosporidium and BRV being the most important pathogens responsible for diarrhea in calves in all cities. Control measures against those pathogens should be enforced to effectively prevent diarrhea in calves in China.

Astigmatism-corrected echelle spectrometer using an off-the-shelf cylindrical lens.

As a special kind of spectrometer with the Czerny-Turner structure, the echelle spectrometer features two-dimensional dispersion, which leads to a complex astigmatic condition. In this work, we propose an optical design of astigmatism-corrected echelle spectrometer using an off-the-shelf cylindrical lens. The mathematical model considering astigmatism introduced by the off-axis mirrors, the echelle grating, and the prism is established. Our solution features simplified calculation and low-cost construction, which is capable of overall compensation of the astigmatism in a wide spectral range (200-600 nm). An optical simulation utilizing ZEMAX software, astigmatism assessment based on Zernike polynomials, and an instrument experiment is implemented to validate the effect of astigmatism correction. The results demonstrated that astigmatism of the echelle spectrometer was corrected to a large extent, and high spectral resolution better than 0.1 nm was achieved.

CD61 promotes the differentiation of canine ADMSCs into PGC-like cells through modulation of TGF-ß signaling.

Previous studies have shown that CD61 (integrin-ß3) promotes the differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into germ-like cells. However, the mechanism remains unclear. In this study, we showed that overexpression of CD61 in canine adipose-derived mesenchymal stem cells (cADMSCs) promotes their differentiation into primordial germ cell (PGC)-like cells. Quantitative real-time PCR, immunocytochemistry and western blot detected higher levels of PGC-specific markers in CD61-overexpressed cADMSCs compared with those in control cells. Moreover, phosphorylation of Smad2, a downstream mediator of transforming growth factor beta (TGF-ß), was increased in CD61-overexpressed cADMSCs than that in control cells. However, the expression of PGC-specific markers was downregulated in cADMSCs treated with a TGF-ß inhibitor. These results suggested that CD61 could induce cADMSCs to differentiate into PGC-like cells by relying on the activation of TGF-ß pathway. ADMSCs possess a considerable potential in treating the infertility of rare animal species.

Primordial germ cell-like cells derived from canine adipose mesenchymal stem cells.

OBJECTIVES: Previous studies have shown that adipose mesenchymal stem cells (AMSCs) share the potency of typical bone marrow mesenchymal stem cells (MSCs); however, there is little information concerning characteristics of canine AMSCs (CAMSCs); it has not previously been made clear whether CAMSCs would be able to differentiate into other cell types. MATERIALS AND METHODS: In this study, typical AMSC lines were established, and their characteristics including morphology, typical markers and differentiation potentiality were tested. RESULTS: The cells exhibited typical MSC morphology and were positive for CD90, CD44 and CD166, considered to be MSCs surface markers. They were negative for CD34 and CD45. The CAMSCs also exhibited embryonic stem cell (ESC) markers, including Oct4 and Sox2, at passage 2. In an appropriate microenvironment, CAMSCs differentiated into EBs and were able to produce cells of the three germ layers. These results indicate that established cells were putative adipocyte-derived MSCs, which also displayed properties of ESCs. Moreover, when the CAMSCs were induced by bone morphogenetic protein 4 (BMP4), they differentiated into PGC-like cells (PGCLCs) and male germ-like cells, which were positive for PR domain-containing 1 (Prdm1), PR domain-containing 14 (Prdm14), doublesex and mab-3 related transcription factor (Dmrt1), as well as for promyelocytic leukaemia zinc finger (Plzf). Quantitative real-time PCR (qRT-PCR) and western blotting analysis verified higher expression levels of these markers. CONCLUSION: This study provides an efficient approach to study germ cell development using CAMSCs.

Sinusoidal phase-modulating fiber-optic interferometer fringe with a feedback control system.

A displacement measurement system using a fiber-optic interferometer fringe projector with a feedback control system is presented and demonstrated. The system utilizes the integrating bucket method to detect the desired phase or the displacement and Fresnel reflection signal to realize measurement of the disturbance and feed it back to the modulated signal of the laser at the same time. The continuous signal truly reflects the error information, as the output light and reflected light share the same optical path. Practical experiments validate the feasibility of this method.

Plasticity of marrow mesenchymal stem cells from human first-trimester fetus: from single-cell clone to neuronal differentiation.

Recent results have shown that bone marrow mesenchymal stem cells (BMSCs) from human first-trimester abortus (hfBMSCs) are closer to embryonic stem cells and perform greater telomerase activity and faster propagation than mid- and late-prophase fetal and adult BMSCs. However, no research has been done on the plasticity of hfBMSCs into neuronal cells using single-cell cloned strains without cell contamination. In this study, we isolated five single cells from hfBMSCs and obtained five single-cell cloned strains, and investigated their biological property and neuronal differentiation potential. We found that four of the five strains showed similar expression profile of surface antigen markers to hfBMSCs, and most of them differentiated into neuron-like cells expressing Nestin, Pax6, Sox1, ß-III Tubulin, NF-L, and NSE under induction. One strain showed different expression profile of surface antigen markers from the four strains and hfBMSCs, and did not differentiate toward neuronal cells. We demonstrated for the first time that some of single-cell cloned strains from hfBMSCs can differentiate into nerve tissue-like cell clusters under induction in vitro, and that the plasticity of each single-cell cloned strain into neuronal cells is different.

Pancreatic islet-like clusters from bone marrow mesenchymal stem cells of human first-trimester abortus can cure streptozocin-induced mouse diabetes.

Bone marrow mesenchymal stem cells (BMSCs) have been reported to possess low immunogenicity and cause immunosuppression of recipients when allografted. They can differentiate into insulin-producing cells and may be a valuable source for islet formation. However, the extremely low differentiating rate of adult BMSCs toward insulin-producing cells and the insufficient insulin secretion of the differentiated BMSCs in vitro prevent their clinical use in diabetes treatment. Little is known about the potential of cell replacement therapy with human BMSCs. Previously, we isolated and identified human first-trimester fetal BMSCs (hfBMSCs). Under a novel four-step induction procedure established in this study, the hfBMSCs effectively differentiated into functional pancreatic islet-like cell clusters that contained 62 ± 14% insulin-producing cells, expressed a broad gene profile related to pancreatic islet ß-cell development, and released high levels of insulin (2.245 ± 0.222 pmol/100 clusters per 30 min) and C-peptide (2.200 ± 0.468 pmol/100 clusters per 30 min) in response to 25 mmol/L glucose stimulus in vitro. The pancreatic islet-like cell clusters normalized the blood glucose level of diabetic model mice for at least 9 weeks when xenografted; blood glucose levels in these mice rose abnormally again when the grafts were removed. Examination of the grafts indicated that the transplanted cells survived in recipients and produced human insulin and C-peptide in situ. These results demonstrate that hfBMSCs derived from a human first-trimester abortus can differentiate into pancreatic islet-like cell clusters following an established four-step induction. The insulin-producing clusters present advantages in cell replacement therapy of type 1 diabetic model mice.