猪只养殖本质就是养肠道。猪只健康生长与肠道菌群平衡密切相关。申请人发现了肠道菌群影响肠道发育、机体代谢组、睾丸代谢组，从而影响精子发生，建立了“肠-睾丸轴”理论，为通过调控肠道菌群、肠道上皮细胞的发育来调控精子发生提供了有力的证据和必要的支持。

1、首次发现仔猪肠道不同类型细胞发育动态变化与肠道微生物、血液代谢组的关系。出生后到断奶前这段时间是仔猪肠道发育的关键时期，而且这段时间仔猪肠道发育是否完善直接关系到成功断奶及断奶后猪只的健康生长和发育。通过高通量单细胞转录组测序、蛋白质组、血液代谢组、及肠道菌群的微生物组，我们研究发现猪小肠发育特点是未分化细胞在逐步增加、肠道吸收细胞先增加后稍微减少到达成熟、分泌细胞时间依赖性地减少，蛋白质组结果证实了这些细胞的发育趋势，而且这些细胞的发育受到转录因子、配体-受体、G蛋白受体、TGF及BMP信号通路的调控。刚出生时小肠粘膜几乎没有菌群存在，随仔猪的生长发育，小肠粘膜的菌群多样性逐步增大，而且有益菌乳酸菌在出生后第3天开始在仔猪回肠粘膜定植，然后一直保持较高含量，而其他一些菌群增加或减少没有太多规律。

2、解析了肠道有益菌群促进肠道上皮细胞发育和优化血液代谢组，从而促进精子发生。猪的健康生长与肠道菌群平衡密切相关，而且肠道菌群影响着精子发生。通过高通量单细胞测序、血液和睾丸代谢组、及肠道菌群的微生物组，我们研究发现有益菌群比如Bacteroidales 和 Lactobacillaceae有利于肠道发育，从而有益于机体代谢组和睾丸代谢组的优化，然后有利于精子发生。利用弱精/少精症病理模型，我们发现病理模型组不仅精子发生受到阻碍（无精症）而且肠道菌群和机体代谢组、睾丸代谢组紊乱，然而有益肠道内容物（菌群）移植可以挽救这种无精症来促进精子发生，进一步研究发现，有益肠道内容物移植可以改善肠道菌群、血液/睾丸代谢组，从而提高生精细胞的活力，改善精子发生，提高了精子的浓度和活力。这项研究在国际上首次证明了肠-睾丸轴的存在，为通过改善肠道来改善精子发生提供了有力的证据。

3、基于肠-睾丸轴，建立肠道菌群修复挽救雄性动物繁殖力的技术方法，实现了动物肠道菌群挽救雄性动物繁殖力。在国际上首次建立了“肠道菌群-雄性繁殖力维持”的新理论。我们发现天然产物褐藻寡糖可以改善雄性动物的繁殖力，主要是通过增加肠道有益菌群，从而改善血液代谢组，然后改善睾丸微环境，进而提高动物的精液质量，提高雄性动物的繁殖力。为了证实肠道菌群在褐藻寡糖改善雄性动物繁殖力中的作用，移植经过褐藻寡糖饲喂后的肠道内容物（菌群）到受体动物体内，然后我们发现受体动物肠道有益菌群增多，血液及睾丸代谢物优化，从而改善了精子发生，提高了精液品质，提高了雄性动物的繁殖力。这是国际上首次发现肠道有益菌群可以改善雄性动物的繁殖力，为以后充分利用改善肠道来改善公猪的繁殖力提供了有力的证据。

1. Pengfei Zhang, Yanni Feng, Lan Li, Wei Ge, Shuai Yu, Yanan Hao, Wei Shen, Xiao Han, Dongxue Ma, Shen Yin, Yu Tian, Lingjiang Min, Zhongyi Sun, Qingyuan Sun, Hongfu Zhang*, Yong Zhao*. Improvement in sperm quality and spermatogenesis following fecal microbiota transplantation from alginate oligosaccharide dosed mice. Gut 2021; 70(1): 222-225. doi: 10.1136/gutjnl-2020-320992. (IF=19.819)

2. Cong Zhang, Bohui Xiong, Liang Chen, Wei Ge, Shen Yin, Yanni Feng, Zhongyi Sun, Qingyuan Sun, Yong Zhao*, Wei Shen*, Hongfu Zhang*. Rescue of male fertility following fecal microbiota transplantation from alginate oligosaccharide dosed mice. Gut 2020; doi: 10.1136/gutjnl-2020-323593. (IF=19.819)

3. Pengfei Zhang, Jing Liu, Bohui Xiong, Cong Zhang, Beining Kang, Yishan Gao, Zengkuan Li, Wei Ge, Shunfeng Cheng, Yanan Hao, Wei Shen, Shuai Yu, Liang Chen, Xiangfang Tang, Yong Zhao*, Hongfu Zhang*. Microbiota from alginate oligosaccharide dosed mice successfully mitigated small intestinal mucositis. Microbiome 2020; 8(1):112. doi:10.1186/s40168-020-00886-x. (IF=11.607)

4. Yong Zhao, Pengfei Zhang, Wei Ge, Yanni Feng, Lan Li, Zhongyi Sun, Hongfu Zhang*, Wei Shen*. Alginate oligosaccharides improve germ cell development and testicular microenvironment to rescue busulfan disrupted spermatogenesis. Theranostics 2020;10(7): 3308-3324. doi: 10.7150/thno.43189. (IF=8.579)

5. Yong Zhao, Yanni Feng, Ming Liu, Liang Chen, Qingshi Meng, Xiangfang Tang, Shukun Wang, Lei Liu, Lan Li, Wei Shen*, Hongfu Zhang*. Single-cell RNA sequencing analysis reveals alginate oligosaccharides preventing chemotherapy- induced mucositis. Mucosal Immunol. 2020; 13(3): 437-448. doi: 10.1038/s41385-019-0248-z. (IF=6.726)

6. Huatao Li, Pengfei Zhang, Yong Zhao*, Hongfu Zhang*. Low Doses of Carbendazim and Chlorothalonil Synergized to Impair Mouse Spermatogenesis through Epigenetic Pathways. Ecotoxicol Environ Saf. 2020;188:109908. doi: 10.1016/j.ecoenv. 2019.109908. (IF=4.872)

7. Xiao Han, Pengfei Zhang, Wei Shen, Yong Zhao*, Hongfu Zhang*. Estrogen receptor-related DNA and histone methylation may be involved in the transgenerational disruption in spermatogenesis by selective toxic chemicals. Front Pharmacol. 2019;10: 1012. (IF=3.845)

8. Pengfei Zhang#, Yong Zhao#, Hongfu Zhang, Jing Liu, Yanni Feng, Shen Yin, Shunfeng Cheng, Xiaofeng Sun, Lingjiang Min, Lan Li, Wei Shen*. Low Dose Chlorothalonil Impairs Mouse Spermatogenesis through the Intertwining of Estrogen Receptor Pathways with Histone and DNA Methylation. Chemosphere 2019;230:384-395. (IF=5.108)

9. Yanan Hao, Hongfu Zhang, Pengfei Zhang, Shuai Yu, Dongxue Ma, Lan Li, Yanni Feng, Lingjiang Min, Wei Shen, Yong Zhao*. Chlorothalonil inhibits mouse ovarian development through endocrine disruption. Toxicol Lett. 2019;303:38-47. (IF=3. 499)

10. Weidong Zhang#, Yong Zhao#, Pengfei Zhang, Yanan Hao, Shuai Yu, Lingjiang Min, Lan Li, Dongxue Ma, Liang Chen, Bao Yi, Xiangfang Tang, Qingshi Meng, Lei Liu, Shukun Wang, Wei Shen, Hongfu Zhang*. Decrease in male mouse fertility by hydrogen sulfide and/or ammonia can be inheritable. Chemosphere 2018;194:147-157. (IF=4.208)