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5. FSH induces apoptosis in follicular granulosa cells of Tianzhu white yak and regulates the expression of Fas/FasL mRNA.

Author

Zhang, Jian, Li, Guyue, Yu, Sijiu, Pan, Yangyang, Fan, Jiangfeng, and Cui, Yan

Subjects
APOPTOSIS inhibition, GRANULOSA cells, CELL morphology, OVARIAN atresia, CELL anatomy, OVARIAN follicle
Abstract

The Tianzhu white yak, a globally rare species, holds immense value as a source for yak materials. While the Fas/FasL pathway is pivotal in granulosa cells apoptosis, its precise molecular workings remain enigmatic. This study endeavours to decipher the role of follicle‐stimulating hormone (FSH) in suppressing ovarian granulosa cells (GC) apoptosis in the Tianzhu white yak. Utilizing advanced cell culture techniques, we employed the MTT method, flow cytometry, fluorescence labelling and RT‐PCR to investigate the apoptotic effects of FSH on yak GCs. Our results reveal that FSH's inhibitory effect on GC apoptosis follows a normal distribution pattern, peaking at an FSH concentration of 100 ng/mL with an apoptosis inhibition rate of 89.31%. When serum was withdrawn, an FSH concentration of 2 × 106 ng/mL reduced apoptosis by 72.84%. Annexin V‐FITC staining revealed membrane invaginations, bubble and protrusion formation on the cell surface, and alterations in membrane structure and cell morphology. Flow cytometry analysis further demonstrated that FSH administration prior to early granulosa cell apoptosis had a more profound effect than during gradual apoptosis, both showing a suppressive effect on early follicular granulosa cell apoptosis. A transcription‐level analysis conducted 3 h prior to serum withdrawal, with the addition of 100 ng/mL FSH, revealed intricate regulations in the expression of Fas/FasL. Notably, we observed a gradual increase in FasL expression over time, yet the presence of FSH effectively down‐regulated FasL expression to baseline levels, without notable changes in Fas expression. Immunocytochemical analysis further confirmed the presence of both Fas and FasL on the cell membrane, nucleus and cytoplasm, with varying intensities depending on the duration of FSH treatment. Our findings suggest that FSH may suppress the apoptotic pathway in follicular primarily by down‐regulating FasL expression, indicating that Fas‐regulated mitochondrial pathways play a more prominent role compared to death receptor pathways. This study offers a fresh perspective on the mechanism underlying follicular atresia in Tianzhu white yaks and lays a solid theoretical foundation for the expansion of this endangered species' population. [ABSTRACT FROM AUTHOR]

Published
2024
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8. LncRNA MEG3 regulates ASK1/JNK axis-mediated apoptosis and autophagy via sponging miR-23a in granulosa cells of yak tertiary follicles

Author

Han, Xiaohong, primary, Pan, Yangyang, additional, Fan, Jiangfeng, additional, Wang, Meng, additional, Wang, Libin, additional, Wang, Jinglei, additional, Afedo, Seth Yaw, additional, Zhao, Ling, additional, Wang, Yaying, additional, Zhao, Tian, additional, Zhang, Tongxiang, additional, Zhang, Rui, additional, Cui, Yan, additional, and Yu, Sijiu, additional

Published
2023
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12. Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing.

Author

Fan Jiangfeng, Luo Yuzhu, Yu Sijiu, Cui Yan, Xu Gengquan, Wang Libin, Pan Yangyang, and He Honghong

Subjects
Medicine, Science
Abstract

Yak milk is superior to common cow milk in nutrients including protein, fat and calories. However, the milk yield of the yak is very much lower compared with other dairy bovines. To understand the molecular mechanisms of lactogenesis, lactation and mammary gland development, mammary tissue samples were taken from five yaks during a dry period (DP, n = 3) and lactation period (LP, n = 2). Two types of cDNA sequence libraries that reflected the different physiological states of the mammary gland were constructed using RNA sequencing technology. After removing reads containing adapters, reads containing poly-N and low-quality reads from the raw data, 45,423,478 to 53,274,976 clean reads were obtained from these libraries. A total of 74.72% to 80.65% of the high-quality sequence reads were uniquely aligned to the BosGru v2.0 yak reference genome. Using the DESeq R package, 360 differentially expressed genes were detected between the two groups when the adjusted P value (padj < 0.05) was used as the cutoff value; this included 192 upregulated and 168 downregulated genes in the yak mammary gland tissue of the DP compared to the LP. A gene ontology analysis revealed that the most enriched GO terms were protein binding, multi-organism process, immune system and others. KEGG pathway analysis indicated that the differentially expressed genes were mostly enriched in Hippo signaling, insulin signaling, steroid biosynthesis and others. The analysis of the up- and downregulated genes provides important insights into the molecular events involved in lactogenesis, lactation and mammary gland development and will guide further research to enhance milk yield and optimize the constituents of yak milk.

Published
2018
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23. FGF10 enhances yak oocyte fertilization competence and subsequent blastocyst quality and regulates the levels of CD9, CD81, DNMT1, and DNMT3B

Author

Pan, Yangyang, primary, Wang, Meng, additional, Baloch, Abdul Rasheed, additional, Zhang, Qiang, additional, Wang, Jinglei, additional, Ma, Rui, additional, Xu, Gengquan, additional, Kashif, Jam, additional, Wang, Libin, additional, Fan, Jiangfeng, additional, Cui, Yan, additional, and Yu, Sijiu, additional

Published
2019
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26. A fully human anti-CD47 blocking antibody with therapeutic potential for cancer

Author

Shuang Wang, Sun Zhiwei, Fan Jiangfeng, Da-di Zeng, Ang Chen, Xiao-peng Yang, Qiang Sun, Peng Du, Lei Xu, Weicai Zhang, and Qiu Weiyi

Subjects
0301 basic medicine, Phage display, Time Factors, Phagocytosis, Mice, Nude, CD47 Antigen, Antibodies, 03 medical and health sciences, Antineoplastic Agents, Immunological, cell-in-cell, Blocking antibody, Medicine, Animals, Humans, CD47, anti-CD47 antibody, Mice, Inbred BALB C, biology, Dose-Response Relationship, Drug, business.industry, Macrophages, Cancer, phagocytosis, U937 Cells, Precursor Cell Lymphoblastic Leukemia-Lymphoma, medicine.disease, Xenograft Model Antitumor Assays, Immune checkpoint, Coculture Techniques, Leukemia, Myeloid, Acute, 030104 developmental biology, HEK293 Cells, Oncology, Cancer cell, Immunology, biology.protein, cancer therapy, Antibody, business, Cell Surface Display Techniques, Research Paper
Abstract

// Dadi Zeng 1, * , Qiang Sun 1, * , Ang Chen 1 , Jiangfeng Fan 1 , Xiaopeng Yang 1 , Lei Xu 1 , Peng Du 1 , Weiyi Qiu 1 , Weicai Zhang 1 , Shuang Wang 1 , Zhiwei Sun 1 1 Beijing Institute of Biotechnology, Fengtai District, Beijing 100071, China * These two authors contributed equally to this work Correspondence to: Zhiwei Sun, email: szwyhhh@aliyun.com Shuang Wang, email: 18910810680@163.com Weicai Zhang, email: drzhangweicai@163.com Keywords: CD47, anti-CD47 antibody, phagocytosis, cancer therapy, cell-in-cell Received: March 17, 2016 Accepted: October 17, 2016 Published: November 15, 2016 ABSTRACT CD47/SIRPα interaction serves as an immune checkpoint for macrophage-mediated phagocytosis. Mouse anti-CD47 blocking antibodies had demonstrated potent efficacy in the treatment of both leukemic and solid tumors in preclinical experimentations, and therefore had moved forward rapidly into clinical trials. However, a fully human blocking antibody, which meets clinical purpose better, has not been reported for CD47 up to date. In this study, we reported the isolation of a fully human anti-CD47 blocking antibody, ZF1, from a phage display library. ZF1 displayed high specificity and affinity for CD47 protein, which were comparable to those for humanized anti-CD47 blocking antibody B6H12. Importantly, ZF1 treatment could induce robust, or even stronger than B6H12, phagocytosis of leukemic cancer cells by macrophage in vitro , and protect BALB/c nude mice from cancer killing by engrafted leukemic cells (CCRF and U937) to a similar extent as B6H12 did. Thus, these data provide primary early pre-clinical support for the development of ZF1 as a fully human blocking antibody to treat human leukemia by targeting CD47 molecule.

Published
2016

27. The expression of Fas/FasL and apoptosis in yak placentomes

Author

Zhao Zhi Wen, Fan JiangFeng, Liu Ben, and Yu Si Jiu

Subjects
medicine.medical_specialty, Fas Ligand Protein, Stromal cell, Placenta, Immunoelectron microscopy, Apoptosis, Biology, Fas ligand, Andrology, Endocrinology, Food Animals, Pregnancy, Internal medicine, medicine, Animals, fas Receptor, Fetus, Trophoblast, General Medicine, Epithelium, medicine.anatomical_structure, Gene Expression Regulation, Giant cell, Cattle, Female, Animal Science and Zoology
Abstract

To clarify the status and distribution of Fas and Fas-Ligand (FasL) in yak's placentomes, immunohistochemistry (IHC) was carried out to analyze the expression and location of Fas and FasL in paraffin embedded sections. The area of positive stained sites was selected and measured using image analyses software (Image Pro-Plus 6.0). So the positive index (PI) was calculated to estimate the intensity of protein expression according to the percentage of positive area in corresponding compartment of the placentomes. In cotyledonary villi, Fas mainly presented on the villous trophoblast cells in early pregnancy. The positive index reached a maximum of 20.7±8.8 at the third month of pregnancy. Then Fas was declined rapidly along with the progress of gestation and the value was 2.8±1.3 after the 7th month of pregnancy. However, in caruncular crypts, Fas was mainly localized to isolated cells or clustered cells of the uterine stroma underlying the caruncular epithelium. The intensity was lower and the positive index was changed between 4.7±0.9 and 8.5±1.6 throughout gestation. For FasL, it gave a distinct immunostained distribution. In cotyledonary villi, FasL was localized dominantly and strongly in the cytoplasm of binuclear, mononuclear and trinuclear trophoblast giant cells (TGC). The positive index of FasL maintained a moderate level all through the gestation. In caruncular crypts, the expression of FasL was weak and the positive index was declined. Only in the first two months, maternal uterine epithelial cells intensely expressed FasL and the index reached to the maximum of 19.8±5.2. The result of subcellular localization of Fas ligand using immunoelectron microscopy technology indicated that FasL was subcellular located in some intracellular vesicles of TGC. This means the vesicles of trophoblast giant cells itself can express FasL. By the TUNEL method, apoptosis was detected in yak placentomes. The amount of apoptotic cells was rare. The fetal chorionic trophoblast cells and caruncular crypt epithelium cells demonstrated higher percentage of apoptosis in middle pregnancy, which suggested that apoptosis plays an important role in placental cellular regeneration. In addition, the apoptosis of maternal caruncular stromal cells provides a local mechanism for maternal immunotolerance to the fetus and this mechanism was mediated by Fas-FasL pathway.

Published
2011

28. Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing

Author

Xu Gengquan, Cui Yan, Pan Yangyang, He HongHong, Fan JiangFeng, Wang Libin, Luo Yu-zhu, and Yu Sijiu

Subjects
0301 basic medicine, Physiology, Molecular biology, Mammary gland, lcsh:Medicine, Gene Expression, Endocrinology, Sequencing techniques, 0302 clinical medicine, Reproductive Physiology, Lactation, Medicine and Health Sciences, lcsh:Science, Multidisciplinary, Gene Ontologies, Reproduction, RNA sequencing, Genomics, Mammary Glands, Body Fluids, Dairying, Milk, medicine.anatomical_structure, Biochemistry, Hippo signaling, 030220 oncology & carcinogenesis, Female, Anatomy, Research Article, Sequence analysis, Mammary Gland Tissue, Steroid biosynthesis, Biology, Beverages, 03 medical and health sciences, Exocrine Glands, Mammary Glands, Animal, Complementary DNA, Genetics, medicine, Animals, Gene Regulation, RNA, Messenger, Nutrition, Endocrine Physiology, Sequence Analysis, RNA, Gene Expression Profiling, lcsh:R, Insulin Signaling, Reproductive System, Biology and Life Sciences, Computational Biology, Genome Analysis, Diet, Research and analysis methods, Gene expression profiling, Molecular biology techniques, 030104 developmental biology, lcsh:Q, Cattle, Transcriptome, Breast Tissue
Abstract

Yak milk is superior to common cow milk in nutrients including protein, fat and calories. However, the milk yield of the yak is very much lower compared with other dairy bovines. To understand the molecular mechanisms of lactogenesis, lactation and mammary gland development, mammary tissue samples were taken from five yaks during a dry period (DP, n = 3) and lactation period (LP, n = 2). Two types of cDNA sequence libraries that reflected the different physiological states of the mammary gland were constructed using RNA sequencing technology. After removing reads containing adapters, reads containing poly-N and low-quality reads from the raw data, 45,423,478 to 53,274,976 clean reads were obtained from these libraries. A total of 74.72% to 80.65% of the high-quality sequence reads were uniquely aligned to the BosGru v2.0 yak reference genome. Using the DESeq R package, 360 differentially expressed genes were detected between the two groups when the adjusted P value (padj < 0.05) was used as the cutoff value; this included 192 upregulated and 168 downregulated genes in the yak mammary gland tissue of the DP compared to the LP. A gene ontology analysis revealed that the most enriched GO terms were protein binding, multi-organism process, immune system and others. KEGG pathway analysis indicated that the differentially expressed genes were mostly enriched in Hippo signaling, insulin signaling, steroid biosynthesis and others. The analysis of the up- and downregulated genes provides important insights into the molecular events involved in lactogenesis, lactation and mammary gland development and will guide further research to enhance milk yield and optimize the constituents of yak milk.

Published
2018

29. Colony-stimulating factor 2 enhances the developmental competence of yak (Poephagus grunniens) preimplantation embryos by modulating the expression of heat shock protein 70 kDa 1A

Author

Wen, Zexing, primary, Pan, Yangyang, additional, Cui, Yan, additional, Peng, Xiumei, additional, Chen, Ping, additional, Fan, Jiangfeng, additional, Li, Guyue, additional, Zhao, Tian, additional, Zhang, Jian, additional, Qin, Shujian, additional, and Yu, Sijiu, additional

Published
2017
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30. A light and electron microscopy study of the yak placentome.

Author

T. Xu, B. Liu, Y. Cui, J. He, J. Fan, S. Yu, Xu, Tao, Liu, Ben, Cui, Yan, He, Junfeng, Fan, Jiangfeng, Yu, Sijiu, Xu, T, Liu, B, Cui, Y, He, J, Fan, J, and Yu, S

Abstract

Background and Materials and methods: In order to clarify and reveal the morphological characteristics of yak placentomes, placentomes obtained from 151 to 180 days of pregnant yaks were observed using light microscopy and scanning and transmission electron microscopy. Results: The results indicated that sessile, dome-shaped yak placentomes seemed to have a relatively complex villous-crypt architecture pattern. There was a straight maternal plate beneath the placentome. Plentiful uterine glands and a dense cellular layer were present in the endometrium lamina propria close to the maternal plate. Trophoblast giant cells appeared to have similar ultrastructure features to these in other ruminants, including abundant mitochondria, an extensive array of rough endoplasmic reticulum, advanced Golgi complex and many specific secretory granules. Trophoblast giant cells could also secrete neutral and acid glycoconjugates. Furthermore, numerous glycoconjugates were distributed in the connective zones between mononuclear trophoblast cells and crypt epithelial cells as well as in maternal connective tissues. Mononucleate trophoblast cells, which had abundant microvilli that interdigitated with the corresponding microvilli arising from the crypt epithelial cells, had numerous mitochondria and vesicles, but there were no glycoconjugates. Conclusions: The morphological structures of yak placentomes were similar to those of other bovid genera; however, certain differences were observed. These findings might provide morphological evidence for evolutionary relationships between different bovid genera. [ABSTRACT FROM AUTHOR]

Published
2019
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