瑶山树鼩皮脂腺瘤诊断及瘤内菌分离鉴定与生物学特性研究

doi:10.16431/j.cnki.1671-7236.2025.10.045

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (10): 5009-5020.doi: 10.16431/j.cnki.1671-7236.2025.10.045

• 临床兽医 • 上一篇

瑶山树鼩皮脂腺瘤诊断及瘤内菌分离鉴定与生物学特性研究

戴颖涵¹, 张达培¹, 李竹欣¹, 翟珊珊¹, 张杰¹, 陶俊宇^1,2, 刘欢^1,3, 梁亮¹, 运晨霞¹, 曹颖颖¹, 徐子恒^1,2, 冷静^1,2,4, 唐海波^1,2

1. 广西中医药大学, 南宁 530200;
2. 广西高发传染病中西医结合转化医学重点实验室, 南宁 530200;
3. 广西中医药防治医学分子生物重点实验室, 南宁 530024;
4. 广西特色实验动物模型重点实验室, 南宁 530200

修回日期:2025-03-19 发布日期:2025-09-30
通讯作者: 徐子恒, 冷静, 唐海波 E-mail:gxuxzh2018@163.com;lj986771558@163.com;tanghb@gxtcmu.edu.cn
作者简介:戴颖涵，E-mail:1023035215@qq.com;张达培，E-mail:405104023@qq.com。
基金资助:
广西自然科学基金项目(2024GXNSFAA010233)；广西中医药防治医学分子生物重点实验室开放课(GXHC-MBTCM-03)；中医药生物新材料关键技术研究与智能装备开发学科交叉创新团队(GZKJ2303)；广西中医药大学高层次人才培育创新团队(2022B006)；广西高校千名中青年骨干教师培育计划(第三期)[桂教师范(2019)81号]

Diagnosis of Sebaceous Adenoma of Tupaia belangeri subsp.yaoshanensis, Isolation,Identification and Biological Characteristics of Intratumoral Bacteria

DAI Yinghan¹, ZHANG Dapei¹, LI Zhuxin¹, ZHAI Shanshan¹, ZHANG Jie¹, TAO Junyu^1,2, LIU Huan^1,3, LIANG Liang¹, YUN Chenxia¹, CAO Yingying¹, XU Ziheng^1,2, LENG Jing^1,2,4, TANG Haibo^1,2

1. Guangxi University of Chinese Medicine, Nanning 530200, China;
2. Guangxi Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine, Nanning 530200, China;
3. Guangxi Health Commission Guangxi Key Laboratory of Molecular Biology of Preventive Medicine of Traditional Chinese Medicine, Nanning 530024, China;
4. Key Laboratory for Complementary and Alternative Medicine Experimental Animal Models of Guangxi, Nanning 530200, China

Revised:2025-03-19 Published:2025-09-30

摘要/Abstract

摘要： 【目的】诊断1例瑶山树鼩自发肿瘤的类型与性质，明确瘤内分离菌株的生物学特性，为细菌与肿瘤之间的关联性提供新的证据。【方法】手术切除肿瘤组织，对肿瘤组织进行病理学及免疫组织化学检测。平板划线法分离获得瘤内菌，进一步对分离菌进行形态观察、生化试验及16S rRNA序列分析。接种昆明小鼠测定分离菌株对小鼠的半数致死量(LD₅₀)，利用药敏纸片法分析该分离株对常用药物的敏感性。【结果】肿瘤位于皮下组织，与脂肪组织相邻，肿瘤组织可见腺泡小叶状结构，外周为基底样细胞，中间可见发育成熟胞浆透亮的皮脂腺细胞，细胞无明显异型性，仅少许核分裂见于基底样细胞。周围脂肪组织液化坏死并融合成大小不一的囊泡状结构，可见反应性多核巨细胞及炎细胞浸润。免疫组织化学显示，肿瘤细胞胞浆中CK14蛋白呈弥漫性强阳性；部分肿瘤细胞胞浆中CK-P蛋白阳性；肿瘤细胞核中P63阳性表达；肿瘤细胞个别细胞核中Ki-67阳性表达，阳性率约为20%。瘤内分离菌为革兰阴性菌，呈杆状。生化鉴定结果显示，分离菌硫化氢、苯丙氨酸、尿素、靛基质、枸橼酸盐均为阳性；侧金盏花醇、山梨醇、丙二酸盐、赖氨酸、鸟氨酸、V-P试验均为阴性，符合普通变形杆菌(Proteus vulgaris)生化特征。16S rRNA测序结果显示分离菌为变形杆菌，与人源豪氏变形杆菌(NR_104767.1)相似性≥99%。该分离菌对昆明小鼠LD₅₀为3.54×10⁶ CFU/0.025 kg。药敏检测结果显示，该分离菌株对头孢类、氨基糖苷类、喹诺酮类等14种药物高度敏感；对阿奇霉素中度敏感；对青霉素类、林可酰胺类等13种药物均有不同程度的耐药。【结论】本研究诊断树鼩自发肿瘤为皮脂腺腺瘤，并于瘤内分离出普通变形杆菌。研究突破了传统认识，为肿瘤微环境中机会致病菌的作用提供了直接证据。该菌株的多重耐药性和高致病性表明其可能通过慢性炎症和免疫逃逸参与肿瘤微环境重塑，但目前证据尚不足以确定其与肿瘤发生发展的直接关系。

关键词: 瑶山树鼩; 皮脂腺瘤; 诊断; 瘤内菌; 普通变形杆菌

Abstract: 【Objective】 This study was conducted to diagnose the type and nature of spontaneous tumors in a Tupaia belangeri subsp.yaoshanensis,clarify the biological characteristics of the strains isolated from the tumor,and provide new evidence for the correlation between bacteria and tumors.【Method】 The tumor was resected surgically and subjected to pathological and immunohistochemical examination.Bacteria within the tumor were isolated using the streak plate method.The isolated bacteria underwent morphological observation,biochemical tests,and 16S rRNA sequence analysis.The median lethal dose (LD₅₀) of the isolated strain to Kunming mice was determined.The drug sensitivity of the isolated strain to commonly used drugs was analyzed using the drug sensitivity paper disc method.【Result】 The tumor was located in the subcutaneous tissue adjacent to fat.It exhibited a lobular structure with basal-like cells peripherally and mature sebaceous cells centrally,showing no significant atypia except for occasional nuclear division in basal-like cells.The surrounding fat tissue showed liquefaction necrosis,forming cystic structures of varying sizes.Reactive multinucleated giant cells and inflammatory cell infiltration were observed.Immunohistochemistry revealed diffuse strong positivity for CK14 in tumor cell cytoplasm.CK-P was positive in some tumor cells,P63 was positive in nucleus,and Ki-67 was positive in individual nucleus with a positivity rate of approximately 20%.The isolated bacteria were Gram-negative rods.Biochemical tests showed that H₂S,phenylalanine,urea,indole and citrate of the isolated strain were positive,and side calendula alcohol,sorbierite,malonate,lysine,ornithine and V-P tests were all negative,which was consistent with the biochemical characteristics of Proteus vulgaris.16S rRNA sequencing identified the isolate as Proteus,and the similarity to human Proteus hauseri (NR_104767.1) was ≥99%.The LD₅₀ of the isolated strain for Kunming mice was 3.54×10⁶ CFU/0.025 kg.Drug sensitivity tests showed that the isolate was high sensitivity to 14 drugs,including cephalosporins,aminoglycosides and quinolones,moderate sensitivity to azithromycin,and varying degrees of resistance to 13 drugs,including penicillins and lincosamides.【Conclusion】 This study diagnosed a spontaneous tumor in Tupaia belangeri subsp.yaoshanensis as a sebaceous adenoma and isolated Proteus vulgaris from the tumor tissue.The findings challenged conventional understanding by providing direct evidence for the role of opportunistic pathogens in the tumor microenvironment.The observed multidrug resistance and high pathogenicity of this bacterial strain suggested its potential involvement in remodeling the tumor microenvironment through chronic inflammation and immune evasion mechanisms.However,current evidence remained insufficient to establish a direct causal relationship between this bacterium and tumor initiation or progression.

Key words: Tupaia belangeri subsp.yaoshanensis; sebaceous gland adenoma; diagnosis; intratumoral bacteria; Proteus vulgaris

中图分类号:

S852.61

戴颖涵, 张达培, 李竹欣, 翟珊珊, 张杰, 陶俊宇, 刘欢, 梁亮, 运晨霞, 曹颖颖, 徐子恒, 冷静, 唐海波. 瑶山树鼩皮脂腺瘤诊断及瘤内菌分离鉴定与生物学特性研究[J]. 中国畜牧兽医, 2025, 52(10): 5009-5020.

DAI Yinghan, ZHANG Dapei, LI Zhuxin, ZHAI Shanshan, ZHANG Jie, TAO Junyu, LIU Huan, LIANG Liang, YUN Chenxia, CAO Yingying, XU Ziheng, LENG Jing, TANG Haibo. Diagnosis of Sebaceous Adenoma of Tupaia belangeri subsp.yaoshanensis, Isolation,Identification and Biological Characteristics of Intratumoral Bacteria[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 5009-5020.

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瑶山树鼩皮脂腺瘤诊断及瘤内菌分离鉴定与生物学特性研究

Diagnosis of Sebaceous Adenoma of Tupaia belangeri subsp.yaoshanensis, Isolation,Identification and Biological Characteristics of Intratumoral Bacteria

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