1.Establishment of a high-precision serial colorful slice image data set of mice
Jun TIAN ; Yue JIA ; Pengfei HE ; Jue GUO ; Rongxiu JIN ; Rui GUO
Chinese Journal of Tissue Engineering Research 2017;21(20):3170-3175
BACKGROUND: With the extensive application of small animal imaging and imaging studies, it requires the investigators to master the cross-sectional anatomy of mice, but the anatomy of mouse nasal cavity and sinuses cannot be obtained through vivisection.OBJECTIVE: To establish a highly precise serial colorful sectional image data set of mice using digital virtual human technology.METHODS: An adult male Balb/c mouse was killed for freezing embedded using modified Kawamoto's method and was then serially scanned at 5 μm of thickness by Leica CM3600XP cryostat microtome. The sectional images were photographed with a 24 million pixels Nikon D750 camera and stored as JPG files. All images were preliminarily registered by manually cropping the images based on the edge of block surface using software photoshop 7.0. Then, the image size, brightness and registration were corrected and adjusted, and the format of the images was unified as TIFF. The data set was imported to Amira 6.0 and was registered again with least square method. The quality of the data set was evaluated by three-dimensional reconstruction. RESULTS AND CONCLUSION: A total of 6034 original images with a size of 87 GB were captured. After cropping, adjusting and registration, the final size was 184 G. The mouse three-dimensional reconstruction model was satisfactory. To conclude, the data set established using the method and system mentioned above is characterized by high resolution and high-fidelity, which might facilitate the further study of the precise anatomy of mice and other fundamental experiments.
2.Research progress of small-scale serial sections-based 3D reconstruction technologies
Yue JIA ; Jun TIAN ; Pengfei HE ; Rongxiu JIN ; Jue GUO ; Rui GUO ; Binquan WANG
International Journal of Biomedical Engineering 2017;40(6):477-481,485
3D reconstruction technologies have been widely used in medical diagnostic assist, surgical simulation, anatomy teaching, etc. The resolution of traditional imaging data such as computed tomography (CT) and magnetic resonance imaging (MRI) cannot meet the needs of life science research. Continuous sectioning technology can provide accurate and high-resolution information for the vast majority of tissues, which make it become a valuable method in the research and exploration of microstructures studies. With the development of microscopy and image processing techniques, significant advances have been made in the small-scale serial sections-based 3D reconstruction technologies. This enables the analysis of sample morphology and tissue function, as well as a more complete assessment of the structure of the tissue. Therefore, this technology has led to profound changes in the fields of cell and tissue biology. In this paper, the common small-scale serial sections-based 3D reconstruction technologies were compared and reviewed.
3.Mechanism of Cangxitongbi Capsules in Mediating Chondrocyte Pyroptosis in Knee Osteoarthritis via Regulating p38 MAPK/NLRP3/Caspase-1 Pathway
Zhimeng ZHANG ; Daotong YUAN ; Ximin JIN ; Rui GONG ; Zhenlong SONG ; Yongkui ZHANG ; Xiaole WANG ; Rongxiu BI ; Wenpeng XIE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):61-68
ObjectiveTo explore the mechanism of Cangxi Tongbi capsules (CXTB) in regulating the p38 mitogen-activated protein kinase (p38 MAPK)/NOD-like receptor protein 3 (NLRP3)/cysteine protease-1 (Caspase-1) signaling pathway to inhibit pyroptosis of cartilage cells in knee osteoarthritis (KOA). MethodSixty male SD rats were randomly divided into a sham operation group, a model group, low, medium, and high dose CXTB groups, and a positive control group, with 10 rats per group. The modified Hulth method was employed to establish a rat model of KOA. According to their respective assignments, rats were administered CXTB (0.25, 0.5, 1.0 g·kg-1) and Celecoxib (24 mg·kg-1) by gavage. The sham operation and model groups were given an equivalent volume of physiological saline. Treatment was performed once daily for 28 days. Micro-computed tomography (Micro-CT) was used to assess bone volume/total volume (BV/TV) and trabecular separation (Tb.Sp). Joint degeneration was evaluated through hematoxylin-eosin (HE) staining, safranin-fast green (SO) staining, and Osteoarthritis Research Society International (OARSI) scoring. Western blot analysis was conducted to measure the expression levels of p38 MAPK, phosphorylated p38 MAPK (p-p38 MAPK), NLRP3, Caspase-1, and gasdermin D (GSDMD) proteins. Real-time PCR was used to assess mRNA expression levels of p38 MAPK, NLRP3, Caspase-1, and GSDMD genes. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum concentrations of inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-18 (IL-18). After knee replacement surgery, cartilage tissue was analyzed using Western blot to assess the protein expression levels of p38 MAPK, p-p38 MAPK, NLRP3, Caspase-1, and GSDMD, and Real-time PCR was used to evaluate gene expression levels of p38 MAPK, NLRP3, Caspase-1, and GSDMD. ResultMicro-CT analysis revealed significant narrowing of the joint space and increased bone spur formation in KOA rats compared with the sham operation group, with a decrease in BV/TV ratio and an increase in Tb.Sp value (P<0.01). Serum levels of TNF-α, IL-1β, and IL-18 were elevated (P<0.01). The protein expression levels of p-p38 MAPK, NLRP3, Caspase-1, and GSDMD in cartilage were significantly increased (P<0.01), and the mRNA expression levels of p38 MAPK, NLRP3, Caspase-1, and GSDMD were also enhanced (P<0.01). Significant differences in protein expression of p-p38 MAPK, NLRP3, Caspase-1, and GSDMD were observed between normal and diseased cartilage tissues after knee replacement surgery (P<0.05), and the gene expression of p38 MAPK, NLRP3, Caspase-1, and GSDMD were also significantly different (P<0.01). HE and SO staining showed roughened joint surfaces, reduced cartilage thickness, and disordered cellular arrangement in KOA rats. OARSI scores were significantly higher (P<0.01). Compared with the model group, treatment with low, medium, and high concentrations of CXTB resulted in increased BV/TV ratios and decreased Tb.Sp values in the knee joints of rats (P<0.01). HE and SO staining indicated a trend towards smoother joint surfaces and reduced OARSI scores (P<0.01). The protein expression levels of p-p38 MAPK, NLRP3, Caspase-1, and GSDMD were notably decreased (P<0.05), as were the mRNA expression levels of p38 MAPK, NLRP3, Caspase-1, and GSDMD (P<0.01). Additionally, serum concentrations of TNF-α, IL-1β, and IL-18 were significantly reduced (P<0.01). ConclusionCXTB intervention may alleviate knee joint degeneration in KOA rats and inhibit the expression of inflammatory factors and pyroptosis of cartilage cells, thereby protecting cartilage. The underlying mechanism may involve modulation of the p38 MAPK/NLRP3/Caspase-1 signaling pathway.
4. Methods and advances in the research of lymphatics in the head and neck
Rui GUO ; Jun TIAN ; Yue JIA ; Pengfei HE ; Rongxiu JIN ; Jue GUO ; Binquan WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2017;52(11):876-880
The lymphatic system plays an important role in human health and disease. In addition to a role in the immune response, the lymphatics can also serve as a pathway for cancer metastasis.Head and neck squamous cell carcinoma (HNSCC) is one of the most common cancers.In the case where the primary focus has been controlled, cervical lymph node metastasis is one of the most important factors affecting the prognosis of head and neck cancer.Thus, understanding the anatomy of the lymphatic system is of paramount importance in predicting cancer metastasis and to perform proper lymph node dissection in cancer patients.The objective of this review is to summarize current imaging approaches that facilitate both basic science and clinical investigations of lymphatic vasculature.Mutation analysis of the laryngeal lymphatic system may provide a theoretical basis for the diagnosis and treatment of HNSCC.