1.Feasibility study of Shuyisha as hemostasis and repair material for liver wound
Jinwei GAO ; Wanshun LIU ; Baoqin HAN ; Jing CHANG ; Yan YANG ; Chenwei FU
Chinese Journal of Trauma 2009;25(7):658-662
Objective To discuss the feasibility of Shuyisha as hemostasis and repair material for liver wound. Methods Hemolysis rate, acute toxicity and eytotoxicity of Shuyisha were measured. A hemorrhage model was established by making an open wound (5 mm× 3 nun ×2 mm) on the left liver lobe of mice. Hemostasis was performed with Shuyisha in experimental group and with Surgicel in control group, when the hemostatic time and total blood loss (TBL) were accurately recorded and regular macro-scopic and histological observation carried out. Results The hemolysis rate of Shuyisha was 2.33%, with maximum tolerance does of over 0.48 g/kg and the eytotoxicity at zero. The hemostatie time of Shuy-isha was (5.00 ±0.00) s, with total blood loss of (0.88±0.18) g/kg, better than Surgicel (P< 0.05). Shuyisha was degraded completely within 14 days, with the wound healed within 21 days in ex-perimental group, much better than Surgieel. Conclusions The hemolysis rate, acute toxicity and cy-totoxicity of Shuyisha are up to the requirement of biomedical materials. Shuyisha has effective hemosta-sis, which may be related to its molecular structure and adhesion.
2.Single-cell RNA sequencing reveals the process of CA19-9 production and dynamics of the immune microenvironment between CA19-9 (+) and CA19-9 (-) PDAC
Deyu ZHANG ; Fang CUI ; Kailian ZHENG ; Wanshun LI ; Yue LIU ; Chang WU ; Lisi PENG ; Zhenghui YANG ; Qianqian CHEN ; Chuanchao XIA ; Shiyu LI ; Zhendong JIN ; Xiaojiang XU ; Gang JIN ; Zhaoshen LI ; Haojie HUANG
Chinese Medical Journal 2024;137(20):2415-2428
Background::Pancreatic ductal adenocarcinoma (PDAC) is one of the main types of malignant tumor of the digestive system, and patient prognosis is affected by difficulties in early diagnosis, poor treatment response, and a high postoperative recurrence rate. Carbohydrate antigen 19-9 (CA19-9) has been widely used as a biomarker for the diagnosis and postoperative follow-up of PDAC patients. Nevertheless, the production mechanism and potential role of CA19-9 in PDAC progression have not yet been elucidated.Methods::We performed single-cell RNA sequencing on six samples pathologically diagnosed as PDAC (three CA19-9-positive and three CA19-9-negative PDAC samples) and two paracarcinoma samples. We also downloaded and integrated PDAC samples (each from three CA19-9-positive and CA19-9-negative patients) from an online database. The dynamics of the proportion and potential function of each cell type were verified through immunofluorescence. Moreover, we built an in vitro coculture cellular model to confirm the potential function of CA19-9. Results::Three subtypes of cancer cells with a high ability to produce CA19-9 were identified by the markers TOP2A, AQP5, and MUC5AC. CA19-9 production bypass was discovered on antigen-presenting cancer-associated fibroblasts (apCAFs). Importantly, the proportion of immature ficolin-1 positive (FCN1+) macrophages was high in the CA19-9-negative group, and the proportion of mature M2-like macrophages was high in the CA19-9-positive group. High proportions of these two macrophage subtypes were associated with an unfavourable clinical prognosis. Further experiments indicated that CA19-9 could facilitate the transformation of M0 macrophages into M2 macrophages in the tumor microenvironment. Conclusions::Our study described CA19-9 production at single-cell resolution and the dynamics of the immune atlas in CA19-9-positive and CA19-9-negative PDAC. CA19-9 could promote M2 polarization of macrophage in the pancreatic tumor microenvironment.
3.Investigation of Mechanism of Qingqiao and Laoqiao on Acute Lung Injury Rats Based on Serum Metabolomics
Kang LI ; Tiantian LYU ; Rongqia CUI ; Shumin XIE ; Shujie GUO ; Wanshun CHANG ; Zhaohua CHEN ; Yanhui ZHU ; Mingsan MIAO ; Haiying HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):89-97
ObjectiveSerum metabolomics of acute lung injury(ALI) in rats was conducted using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) to explore the similarities and differences in the mechanism of Qingqiao(harvested when the fruits of Forsythiae Fructus were initially ripe and still green in color) and Laoqiao(harvested when the fruits of Forsythiae Fructus were ripe) in the treatment of ALI. MethodA total of 24 SD male rats were acclimatized and fed for 1 week, 6 of them were randomly selected for the blank group and 18 for the experimental group. The ALI model was induced in the experimental group by tracheal intubation with lipopolysaccharide(LPS). After successfully constructing the ALI model, these rats was randomly divided into model group, Qingqiao group and Laoqiao group, with 6 rats in each group. The Qingqiao and Laoqiao groups were administered orally once a day at a dose of 1.5 g·kg-1, while the blank and model groups received an equivalent volume of saline for 3 consecutive days. The pathological conditions of rat lung tissues were comprehensively assessed by hematoxylin-eosin(HE) staining, wet-to-dry mass ratio(W/D) of lung tissues, and protein concentration in rat bronchoalveolar lavage fluid(BALF). The levels of interleukin(IL)-6, IL-1β and tumor necrosis factor(TNF)-α in BALF were quantified using enzyme-linked immunosorbent assay(ELISA). UPLC-Q-TOF-MS was used to identify and analyze the chemical compositions of Qingqiao and Laoqiao, and serum metabolomics of rats in each group was analyzed, combined with multivariate statistical analysis with variable importance in the projection(VIP) value>1, P<0.05 from t-test, and fold change(FC)≥1.5 or FC≤0.5 to screen the differential metabolites Qingqiao and Laoqiao for the treatment of ALI. The Kyoto Encyclopedia of Genes and Genomes(KEGG) database was used in combination with MetaboAnalyst for the metabolic pathway analysis of the screened differential metabolites. ResultCompared with the blank group, rats in the model group exhibited enlarged alveolar lumen, ruptured alveoli, interstitial hemorrhage, bronchial exudation of a large number of neutrophils and erythrocytes, and a significant increase in the protein concentration in the BALF and the W/D value of the lung tissues(P<0.01). In contrast, compared with the model group, rats in the Qingqiao group and the Laoqiao group showed reduced bronchial hemorrhage in the lungs, and the protein concentration in the BALF and the W/D value of the lung tissues were significantly decreased(P<0.01), the lung injury was significantly alleviated, but more obvious in the Qingqiao group. Compared with the blank group, the expression levels of IL-6, IL-1β and TNF-α in the BALF of the model group were significantly higher(P<0.01). Additionally, compared with the model group, the expression levels of IL-6, IL-1β and TNF-α in the Qingqiao and Laoqiao groups were significantly lower(P<0.01). The chemical composition analysis of Qingqiao and Laoqiao revealed that 63 components were detected in Qingqiao and 55 components were detected in Laoqiao, with 47 common components, 16 components unique to Qingqiao and 8 components unique to Laoqiao. Characterizing the differences in serum metabolomics in rats, 19 and 12 metabolites were called back by Qingqiao and Laoqiao, respectively. The metabolic pathway enrichment analysis showed that Qingqiao exerted its therapeutic effects by affecting 6 key metabolic pathways, including linoleic acid metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, glycerophospholipid metabolism, α-linolenic acid metabolism, and arachidonic acid metabolism, and Laoqiao exerted therapeutic effects by affecting 6 key metabolic pathways, including linoleic acid metabolism, arachidonic acid metabolism, sphingolipid metabolism, phenylalanine metabolism, ascorbate and aldarate metabolism, and glycerophospholipid metabolism. ConclusionQingqiao and Laoqiao have therapeutic effects on ALI, and Qingqiao is more effective. Both of them can play a therapeutic role in ALI by regulating amino acid metabolism and lipid metabolism, but the metabolic pathways affected by them are different.