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The yak is an agricultural animal with strong disease resistance in Qinghai-Tibet Plateau. Immune organs are directly involved in the body's immune response and protect it from external aggression. In this study, we characterized and evaluated the main markers of interleukin (IL)-1β, IL-17a, hypoxia inducer factor-1 (HIF-1)α, and heat shock protein 90 (HSP90) in the lymph nodes, spleen, thymus, and hemal nodes of adult yaks using network informatics, molecular cloning, immunohistochemistry, real-time quantitative polymerase chain reaction (RT-qPCR), and western blotting. We first cloned the IL-1β and IL-17a mRNA of yaks. A significant feature was the higher IL-1β and IL-17a expression in the lymph nodes than in the spleen, hemal nodes, and thymus. Immunohistochemistry and immunofluorescence revealed that IL-1β and IL-17a cells were mainly located in the paracortex area of the lymph nodes and the T-cell-dependent area in the hemal nodes and spleen. Several HIF-1α proteins were detected in the cortex of the hemal nodes mantle, while HSP90 was detected in the lymphoid nodules of the hemal nodes and lymph nodes. This study sheds light on the relationship between the morphology and function of these organs and provides an important reference for studies on the participation of yak immune organs in immune responses.

Apoptosis and autophagy in granulosa cells (GCs) are highly related to follicular development and atresia. LncRNA MEG3, miR-23a, and apoptosis signal-regulating kinase 1 (ASK-1) have also been reported to be associated with apoptosis and autophagy. However, their relationship to follicular development and the extent to which follicle stimulating hormone (FSH) or luteinizing hormone (LH) can regulate this process remains unknown in yak GCs. Here, the effect of LncRNA MEG3 / miR-23a on apoptosis and autophagy via ASK1 (Apoptosis signal-regulating kinase 1) / JNK (c-Jun N-terminal kinase) in yak GCs were studied. Overexpressing LncRNA MEG3 reduced miR-23a levels and p-967 protein expression, but enhanced ASK1 and JNK mRNA levels as well as t-ASK1, p-845, t-JNK, and p-JNK proteins levels, and promoted apoptosis while attenuating autophagy. Then, the interaction between LncRNA MEG and miR-23a and both miR-23a and ASK1 via dual-luciferase reporter assay was also confirmed. The relationship between LncRNA MEG3 and miR-23a was observed as a negative feedback regulation for ASK1/JNK axis via pcDNA3.1-MEG3 or miR-23a transfection which mimicked alone or in co-transfection. In addition, FSH (10 μg/mL) or LH (100 μg/mL) ability to reverse the effects of LncRNA MEG3 on miR-23a levels and ASK1/JNK axis-mediated apoptosis and autophagy was verified in yak GCs. This is significantly beneficial for decreasing abnormal follicular atresia.

Erythropoietin (EPO), a central protein of erythropoiesis, plays an important role during hypoxia adaptation and is regulated by hypoxia-inducible factor (HIF). However, there is no report on EPO-producing cells and their regulatory mechanisms in yak (Bos grunniens). To understand EPO production and regulation of yak, kidneys from different age of yak were collected and expression of EPO, hypoxia-inducible factor 1 alpha (HIF-1α), and hypoxia-inducible factor 2 alpha (HIF-2α) were detected. Then renal tubule epithelial cells (RTECs) and peritubular interstitial fibroblast-like (RIFs) cells were isolated and cultured to determine their EPO production abilities. Subsequently, the cells were treated with dimethyloxalylglycine (DMOG) and Geldanamycin (GA), which are inhibitors of prolyl-4-hydroxylase domain (PHD) and heat shock protein 90 (HSP90) respectively, and siRNAs of HIF-1α and HIF-2α to explore their effect on EPO production and regulation. The results showed that expressions of EPO, HIF-1α, and HIF-2α were different in the different age groups of yak. High DMOG concentration caused a corresponding increase in the levels of HIF-1α and HIF-2α in RIFs and RTECs, however, EPO levels increased in RIFs only and was not detected at any concentration in RTECs; suggesting that EPO was produced in RIFs. Upon treating RIFs with siRNAs of HIF-1α and HIF-2α, we found that EPO was regulated by PHD through HIF-2α. In addition, increasing GA concentration caused a decrease in expression of HSP90, HIF-1α, HIF-2α, and EPO in RIFs. In conclusion, these findings support our proposition that PHD regulates EPO via HIF-2α in yak RIFs, while HSP90 impelled EPO expression.

Phytochemical investigation of the whole plants of Dicranostigma leptopodum (Maxim) Fedde has lead to the isolation of five alkaloids: dicranostigmine (1), isocorydine (2), corydine (3), protopine (4) and sinoacutine (5). Oxomorphine alkaloid (5) was isolated from the genus Dicranostigma for the first time and the new compound structure (1) was elucidated by various spectroscopic methods including 2D NMR techniques (gCOSY, HMQC, HMBC and NOESY) and HRESI-MS.

The culture of primary Sertoli cells has become an important resource in the study of their function. However, their use is limited because of contamination of isolated cells with other testicular cells, mainly germ cells. The aim was to establish technique to obtain pure yak Sertoli cells as well as to study the growth kinetics and biological characteristics of Sertoli cells in vitro. Two-step enzyme digestion was used to separate and culture yak Sertoli cells. Cultured using starvation method and the hypotonic treatment were also invented to get pure yak Sertoli cells. Furthermore, the purification of Yak Sertoli cells were identified according to their characteristics, such as bipolar corpuscular around the nucleus and expression of Fasl, in addition to their morphology. The average viability of the Sertoli cells was 97% before freezing and 94.5% after thawing, indicating that cryopreservation in liquid nitrogen had little influence on the viability of Sertoli cells. The growth tendency of yak Sertoli cells was similar to an S-shaped growth curve. Purified yak Sertoli cells frequently exhibited bipolar corpuscula in nucleus after Feulgen staining, and did have a positive reaction of Fasl by the immunocytochemical identification. After recovery chromosomal analysis of Sertoli cells had a normal chromosomal number of 60, comprising 29 pairs of autosomes and one pair of sex chromosomes. Assays for bacteria, fungi and mycoplasmas were negative. In conclusion, yak Sertoli cells have been successfully purified and cultured in vitro, and maintain stable biological characteristics after thawing. Therefore, it will not only preserve the genetic resources of yaks at the cellular level, but also provide valuable materials for transgenic research and feeder layer and nuclear donor cells in yak somatic cell cloning technology.

To isolate and purify beta-defensin from the neutrophils of Yak.The method of percoll gradient centrifugation was employed to purify neutrophils from Yak peripheral blood. The crude extraction from Yak neutrophils was isolated respectively through a way of extraction by 50 mL/L acetic acid, acid soluble extract of Yak neutrophils was obtained. Then it was further purified by Bio-Gel P-10 Polyacrylamide gel filtration and RP-HPLC, and 9 out of 22 apices showed antibacterial activity. 9 apices showing antibacterial activity were selected to be analyzed by mass spectrograph.The antibacterial activities of the extracts from every step were analyzed by agarose radial diffusion assay.The molecular weight of the purified Yak BNBD-1-3 was determined to be 4.2, 4.6 and 4.8 kDa by mass spectrograph. BNBD-1-3 from Yak neutrophils were able to effectively kill Escherichia coli, Staphylococcus aureus and Bacillus subtilis.Yak neutrophils have beta-defensin which have broad-spectrum antimicrobial activity, Yak defensin has an important part in innate immunity.