白消安处理消融猪内源精原干细胞的效果及外源精原干细胞移植

目的 研究猪睾丸组织注射白消安消融猪内源精原干细胞(Spermatogonial stem cell,SSC)的效果,以及猪SSC同种异体移植后对内源性SSC消融受体生殖能力恢复的影响。方法 采用3 mg/kg剂量的白消安对9头6周龄大白公猪进行睾丸注射,另外3头注射2 mL 二甲基亚砜作为对照。3周后,对试验组公猪以相同剂量进行第2次睾丸注射。第2次注射3周后,采集试验组和对照组公猪睾丸进行相关检测,评估内源性SSC消融情况。第2次白消安注射1个月后,以两步酶消化法处理5~7日龄大白仔猪睾丸分离得到睾丸单细胞悬液,并用明胶差速贴壁法进行纯化,纯化后以免疫荧光和流式细胞术分析SSC的纯度。异体移植SSC 4个月后,用微卫星标记检测受体公猪精液以及睾丸组织中供体来源SSC的存在。结果 以3 mg/kg剂量的白消安处理大白公猪2次后,睾丸组织苏木精−伊红染色以及免疫组织化学染色结果显示,试验组睾丸曲细精管中各级生精细胞消融但其支持细胞结构完好,可以支持外源性SSC的定植及发育。免疫荧光以及流式细胞术结果表明,分离得到的睾丸单细胞悬液经纯化后UCHL-1阳性细胞占比由差速贴壁前的16.3%提高到了50.8%。苏木精−伊红染色以及免疫组织化学染色结果显示,猪SSC移植4个月后,移植组睾丸组织生精细胞恢复,在受体睾丸曲细精管基底膜上可检测到UCHL-1阳性SSC。受体睾丸组织的微卫星标记分析显示了供体SSC的存在,表明移植进入受体睾丸中的供体SSC可以在受体睾丸中定植存活超过4个月;精液微卫星标记未检测到供体来源的精子。结论 以3 mg/kg剂量的白消安注射公猪睾丸能有效消融内源性SSC,可以用来制备SSC移植的受体猪。两步酶消化及明胶差速贴壁法可成功分离纯化猪SSC。猪SSC经同种异体移植后可以在受体睾丸中定植存活超过4个月。     

麦瘟病与小麦抗麦瘟基因研究进展

小麦麦瘟病,由梨孢菌小麦专化型(Magnaporthe oryzae, sp. Triticum, MoT) 侵染所致,最早于1985年在巴西发现,随后传播到其他南美国家。2016与2017年,麦瘟病登陆亚洲,相继在孟加拉国和印度暴发,对亚洲乃至全世界的小麦生产形成了潜在威胁。近年来,世界各国的科学家对麦瘟病进行了广泛和深入的研究,取得了较大进展。本文对小麦麦瘟病病原菌、侵染过程、流行因素和小麦抗病基因的研究进展进行了综述,并介绍了小麦麦瘟病抗性评价的方法。本研究组研究发现,在人工接种条件下,水稻来源的稻瘟病菌株(MoO)可侵染小麦引起典型麦瘟症状。笔者认为,虽然目前我国还没有麦瘟病发生的报道,但部分地区存在麦瘟病的发病条件,随着全球气候变暖麦瘟病有暴发的可能,需引起高度重视。建议加快麦瘟病发病机理及小麦抗性基因发掘的研究,以提升我国麦瘟病的研究水平,并为预防麦瘟病的发生未雨绸缪。     

Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

华蟾毒精对胃腺癌细胞MGC-803作用的研究

研究华蟾毒精（cinobufagin,CBG）体外对人胃腺癌细胞MGC-803的抑制作用,并初步探讨其抗肿瘤的作用机制。MTT法测定CBG对胃腺癌MGC-803细胞的生长抑制情况;采用光镜观察CBG对人胃腺癌细胞MGC-803形态的影响;流式细胞术检测CBG对MGC-803细胞的抑制作用、细胞周期及细胞早期凋亡的影响。一定浓度范围内CBG对MGC-803细胞的增殖有剂量、时间依赖性抑制作用;光镜下可见细胞形态明显发生变化,细胞变圆,分泌颗粒增多,细胞折光性下降,大部分细胞脱壁;流式细胞仪检测MGC-803细胞,G1期细胞减少,大量细胞停滞在G2/M期,凋亡细胞增多。CBG对MGC-803细胞的抑制作用与诱导其细胞阻滞及凋亡密切相关。     

一例细菌对MDCK细胞污染的检测

MDCK细胞培养液（含双抗）表面有一层薄的油状膜，培养液不混浊。细菌学检测为一种球菌污染。对污染菌株进行药敏试验。使用MEM（含双抗）多次清洗细胞培养瓶及细胞表面，细胞培养液中改加菌株敏感的恩诺沙星。细菌学检测结果表明，加入恩诺沙星有效地控制和清除了细胞中污染的该株细菌。     

Cellular localization of Visudyne® as a function of time after local injection in an in vivo model of squamous cell carcinoma: an investigation into tumor cell death

Objective To evaluate the effects of time on cellular localization of Visudyne^® after local injection. Animals Twenty athymic nude mice. Procedures A squamous cell carcinoma (SCC) cell line (A‐431) was injected into right and left dorsolumbar subcutaneous tissue of each mouse, representing treatment (T) and control (C) tumors. In experiment 1 (Exp 1; n = 10) and 2 (Exp 2; n = 10), the T tumors received a local injection of Visudyne^® (0.1 mg/cm³), and C tumors received an equal dose of 5% dextrose in water (D5W). Mice were randomly subdivided into two groups (A and B; n = 5 per group). Mice in Exp 1A and B were sacrificed 1 and 30 min after local injection, respectively. Experiment 1A and B tumors were evaluated by fluorescence microscopy to determine drug localization. Experiment 2A and B tumors were exposed to LED illumination 1 and 30 min after injection, respectively, and evaluated by transmission electron microscopy (TEM) to determine ultrastructural tumor cell damage. Results Fluorescence was detected within the cytoplasm of T tumors in both Exp 1A and B. Significance was detected in fluorescence intensity between T1 min vs. T30 min (P = 0.03) and between T1 min and C1 min tumors (P = 0.01), respectively. Tumors in Exp 2A and B demonstrated evidence of apoptotic cell death. Conclusions Fluorescence microscopy demonstrated higher Visudyne^® concentration within SCC cytoplasm of 1 min compared with 30‐min tumors. Transmission electron microscopy results revealed that tumors treated by photodynamic therapy (PDT) within 30 min of local injection undergo cellular apoptosis.     

酵母壁多糖对断奶仔猪肠道挥发性脂肪酸和微生物菌群的影响

本研究旨在探讨饲粮中添加酵母壁多糖对断奶仔猪肠道挥发性脂肪酸和微生物菌群的影响。试验采用单因素试验设计方法,选取180头遗传背景一致、健康状况良好、胎次和体重接近的21日龄断奶仔猪,随机分为4个组,每组5个重复,每个重复9头猪。4个组试验猪分别饲喂对照饲粮(未添加酵母壁多糖)、0.15%酵母壁多糖饲粮、0.30%酵母壁多糖饲粮和0.45%酵母壁多糖饲粮。试验期21 d。结果表明:1)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著提高了仔猪结肠乙酸的含量(P0.05);其中,0.30%和0.45%酵母壁多糖还显著提高了结肠丙酸、丁酸以及总挥发性脂肪酸的含量(P0.05),且二者之间差异不显著(P0.05)。2)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著降低了盲肠沙门氏菌和大肠杆菌数量(P0.05),且0.30%和0.45%酵母壁多糖组之间差异不显著(P0.05)。由此可知,酵母壁多糖可提高仔猪肠道挥发性脂肪酸含量,并改善肠道微生物菌群结构,根据回归方程预测,酵母壁多糖在仔猪饲粮中的适宜添加水平为0.31%~0.40%。     

意蜂采用2,3日龄工蜂幼虫产浆的研究

对意蜂采用２和３日龄工蜂幼虫产浆的王台接受率和不同时间取浆浆量进行了研究。研究结果显示：①２日龄工蜂幼虫的王台接受率（８２．４２％）与采用３日龄幼虫的（８１．２１％）无显著差异;②２和３日龄幼虫产浆的台平均产浆量最高峰分别在各自移虫后第６０小时和第４８小时,浆量较高时段分别在移虫后上时和第４２－５４小时;③２日龄幼虫产浆最高时段台平产浆量（０．３７５４ｇ）极显著高于３日龄的（０．３１５７ｇ）但采用