1.Expression of thymidine phosphorylase in human metastatic liver cancer tissues from colorectal cancer
Wei YOU ; Jimin ZHANG ; Xiangcai ZOU ; Zhuofang HAO ; Degui LIAO ; Shizhang HUANG ; Chuyuan HONG
International Journal of Surgery 2009;36(7):440-444
Objective To detect the tbymidine pbospborylnse (TP) expression in metastatic liver cancer tissues from human colorectal cancer by immunohistochemistry, and analyze the correlation between TP ex-pression and the tumor-associated macrophages (TAM), and the prognosis of patients. Methods Twenty-eight metastatic liver cancer specimens resected from patients with colorectsl cancer, were immunohistochem-ically stained by 654-1, an anti-TP monoclonal antibody, IC6-203, another anti-TP monoclonal antibody, PG-M1, anti-macrophage marker CD68 monoclonal antibody. Morphometrical analysis and positive cell counting were performed, and the correlation of TP expression with the patient's prognosis was evaluated. Results In normal liver tissues, the hepatic cells apart from cancer nests were weakly positive for 654-1 as well as for 1C6-203. The most TP-positive cells were distributed mainly along the invasive margin of cancer or around the cancer nests. In the corresponding areas, CD68-positive macrophages were also increased. The distribution patterns of CD68-positive cells were similar to those of TP-pesitive cells. The numbers of the TP-positive cells stained by 654-1 were significantly correlated with numbers of 1C6-203 positive cells (r=0.697, P<0.01), also correlated with the numbors of CD68-positive cells (r=0.703, P<0.01). While the numbers of 1C6-203 positive cells had no significant differences with the numbers of CD68-positive cells (r=0.359, P>0.05). The TP-pesitive cancer cells both for 654-1 and for 1C6-203 were detected only in 2 of 28 specimens. Both the number of TP-pesitive cells for 654-1 and 1C6-203, and the number of CD68-positive cells had no correlation with the survival period of patients. Conclusions In the metastatic liver cancer tissues of human colorectsl cancer, the TP-expreasinn stained by 654-1 was coincidence with 1C6-203, and the most important source of TP-expreasion is the TAM in stromal tissues around cancer nests, while the cancer cells are little expressed. The numbers of TP-positive cells stained by 654-1 are significantly related with CD68-pesitive macrophages, but not with the post-operation survival period of patients.
2.The expression of PIWI protein in tissue of human colon cancer and its clinical significance
Juan LI ; Xiaoying SHEN ; Shizhang ZHU ; Wei WANG ; Ximei CHEN ; Changqing YANG ; Hengjun GAO
Chinese Journal of Digestion 2011;31(4):243-248
Objective To study the expression of PIWIL1, PIWIL3 and PIWIL4 in human colon cancer and its clinical significance. Methods We collected cancerous tissues and its adjacent tissues of 106 patients with colon cancer, two tissue microarrays were constructed, with 62 and 150 points respectively. We studied the expression of PIWIL1, PIWIL3 and PIWIL4 through immunohistochemistry. Results The expression of PIWIL1, PIWIL3 and PIWIL4 were significantly higher in cancerous tissues than those in adjacent tissues (P<0. 01). In cancerous tissues and its adjacent tissues, postive correlation were seen among PIWIL1, PIWIL3 and PIWIL4 expression (P<0. 01). PIWIL1 expression was significant higher in low differentiation group than that in high differentiation group (t =- 2. 840, P<0.01 ). PIWIL3 expression was higher in high clinical stage than that in low clinical stage (F= 3. 112, P<0.05). The expression of PIWIL3 and PIWIL4 were significantly higher in patients with colon cancer with distant metastasis than those without distant metastasis (t= -3. 349, P<0.01 ; t = - 2. 168, P<0. 05). PIWIL3 and PIWIL4 expression were correlated with occurring of colon cancer (P<0. 01). Conclusions The expressions of PIWIL1,PIWIL3 and PIWIL4 in colon cancer were correlated with the differentiation, clinical stage and distant metastasis of colon cancer. PIWIL3 and PIWIL4 expression were two independent related factors of occurring of colon cancer, which would be furtherly investigated to be served as novel markers for early diagnosis and promising molecular targets for colon cancer therapy.
3.Involvement of fascin-1-mediated autophagy in the biological behavioral of endometrial cells.
Xiaomei LUO ; Wei CHENG ; Shizhang WANG ; Zhihong CHEN
Journal of Central South University(Medical Sciences) 2018;43(9):957-963
To explore the mechanism for the role of autophagy in endometriosis, and to provide a theoretical basis for prevention and treatment of endometriosis.
Methods: The endometrial CRL-7566 cells were treated with ATG5 siRNA, autophagic activator rapamycin and autophagic inhibitor 3-MA, respectively. The cell proliferation and invasion were detected by clonal formation, cell growth curve and MTT assay. The clinical specimens of endometriosis were collected from 20 cases. The expression of autophagy marker LC3II and autophagy substrate protein P62 were detected.
Results: Rapamycin inhibited the proliferation and clonal formation of CRL-7566 cells, while autophagy inhibitor 3-MA and ATG5 siRNA showed opposite effect. Moreover, rapamycin inhibited filopodia growth in endometriosis, whereas overexpression of filopodia-relevant protein fascin-1 inhibited the decrease in invasiveness caused by rapamycin. In clinical samples, we also found a significant decrease of LC3II while an increase in P62 compared with the control group.
Conclusion: Autophagy inhibition may contribute to an increase in endometrial cell proliferation and invasiveness. Autophagy activation could be a potential strategy for endometriosis therapy.
Autophagy
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drug effects
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genetics
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Carrier Proteins
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genetics
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metabolism
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Cell Line
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Cell Proliferation
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drug effects
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Endometriosis
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physiopathology
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Endometrium
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cytology
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Female
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Gene Expression Regulation
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Humans
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Microfilament Proteins
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genetics
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metabolism
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Microtubule-Associated Proteins
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genetics
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RNA-Binding Proteins
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genetics
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Sirolimus
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pharmacology
4. Effect of helicid on osteoarthritis of joint instability model and its mechanism
Ge-Ge LIU ; Ran LIU ; Ben-Feng QIU ; Xue-Jun HE ; Xin-Yan CHEN ; Yun-Zhe HUANG ; Yuan-Wei JIA ; Shizhang LING ; Jie SHEN ; Shizhang LING
Chinese Pharmacological Bulletin 2023;39(8):1457-1463
Aim To study the therapeutic effect of helicid on osteoarthritis (OA) of joint instability model, and explore the mechanism of helicid in the treatment of OA. Methods A rat knee model of OA was established by the medial meniscectomy (MMx) method. After treatment with helicid, HE and safranin O/fast green staining methods were used to observe the his-topathological changes of rat knee articular cartilage; Western blot was used to detect the protein expression level of Trpvl in rat synovial tissue. Immunohistochemical staining was used to examine the expression of Trpvl in rat knee articular cartilage and synovial tissues. Results Helicid significantly slowed down the degeneration of rat knee articular cartilage as shown by HE and safranin O/fast green staining. Western blot results showed that helicid down-regulated the expression of Trpvl in rat synovial tissue examined. Immunohistochemical results showed that helicid significantly reduced the expression of Trpvl in both of knee articular cartilage and synovial tissues. Conclusions Helicid prominently decreases MMx-induced articular cartilage damage and cartilage matrix loss, thereby exerting a therapeutic effect on OA.
5.Ameliorative Effect and Mechanism of Qingwen Baiduyin on Lipopolysaccharide-induced Acute Lung Injury
Ju YANG ; Zhao ZHANG ; Jiawei WANG ; Shuying XIE ; Pengyan LI ; Liwei LANG ; Shizhang WEI ; Haotian LI ; Manyi JING ; Yanling ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(15):1-13
To investigate the therapeutic effect and mechanism of Qingwen Baiduyin on acute lung injury (ALI) in mice induced by lipopolysaccharide (LPS). MethodA total of 144 C57BL/6 mice were randomly divided into the following groups: a normal group, a model group (LPS, 5 mg·kg-1), a dexamethasone group (5 mg·kg-1), and low-, medium-, and high-dose Qingwen Baiduyin groups (14.105, 28.21, 56.42 g·kg-1). The mice were treated once daily for 5 days. One hour after the final administration, the ALI model was established by intratracheal instillation of LPS, and samples were collected at 6 h and 24 h after modeling. The arterial blood gas index of mice was analyzed. The total protein content, total cell count, Evans blue dye (EBD) content, and lung tissue wet-to-dry weight ratio (W/D) of bronchoalveolar lavage fluid (BALF) were measured. Hematoxylin-eosin (HE) staining was performed to assess the pathological changes in mouse lung tissue. Western blot was used to detect the expression levels of key proteins in the Janus kinase 1/signal transducer and activator of transcription 1/interferon regulatory factor 1 (JAK1/STAT1/IRF1) signaling pathway in lung tissue. ResultCompared with the normal group, the model group showed reduced arterial oxygen pressure (pO2), oxygen saturation (SO2), and lung tissue W/D (P<0.05, P<0.01), increased carbon dioxide pressure (pCO2), total protein content, total cell count, EBD content, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), chemokine CXC ligand 1 (CXCL1), chemokine CXC ligand 2 (CXCL2), chemokine CXC ligand 9 (CXCL9), and chemokine CXC ligand 10 (CXCL10) content (P<0.05, P<0.01), thickening of the alveolar walls, fusion of alveolar cavities, and infiltration of inflammatory cells in lung tissue, increased proportion of M1 macrophage polarization and lung cell apoptosis (P<0.05), and increased protein expression levels of JAK1, phosphorylated JAK1 (p-JAK1), inducible nitric oxide synthase (iNOS), STAT1, phosphorylated STAT1 (p-STAT1), IRF1, gasdermin D (GSDMD), and mixed lineage kinase domain-like protein (MLKL) (P<0.05, P<0.01). Compared with the model group, Qingwen Baiduyin significantly increased pO2, SO2, and lung tissue W/D (P<0.05, P<0.01), improved the pathological changes in lung tissue, and reduced pCO2, total protein content, total cell count, EBD content, IFN-γ, TNF-α, IL-1β, CXCL1, CXCL2, CXCL9, and CXCL10 content, proportion of M1 macrophage polarization, and protein expression levels of JAK1, p-JAK1, iNOS, STAT1, p-STAT1, IRF1, GSDMD, and MLKL (P<0.05, P<0.01). ConclusionQingwen Baiduyin can improve the lung inflammatory response and reduce lung cell apoptosis in mice with ALI by inhibiting the JAK1/STAT1/IRF1 signaling pathway, thereby exerting a lung-protective effect.
6.Effect and Mechanism of Zuojinwan on DSS-induced Ulcerative Colitis
Zhao ZHANG ; Ju YANG ; Jiawei WANG ; Shuying XIE ; Wenbin WU ; Manyi JING ; Liwei LANG ; Haotian LI ; Shizhang WEI ; Jian WANG ; Yanling ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(16):1-11
ObjectiveTo explore the effect and mechanism of Zuojinwan (ZJW) in the treatment of ulcerative colitis (UC) through network pharmacology and experimental validation. MethodUsing network pharmacology and molecular docking, the active components and potential mechanism of ZJW in treating UC were preliminarily identified. Forty-eight male C57BL/6J mice were randomly divided into a normal group, a model group, a sulfasalazine group (300 mg·kg-1), and low-, medium-, and high-dose ZJW groups (1.82, 3.64, 7.28 g·kg-1). The UC model was induced by dextran sulfate sodium (DSS), and oral administration of drugs began on the third day of modeling, lasting for 7 days. The general condition of mice was observed daily, and the disease activity index (DAI) was evaluated. Hematoxylin-eosin (HE) staining was performed to observe histopathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10 in mouse serum. The molecular mechanism was validated using Western blot. ResultNetwork pharmacology predicted that the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway might be a key pathway in the regulation of UC by ZJW. Molecular docking results showed good binding ability between the key components of ZJW and core targets. Animal experiment results showed that compared with the normal group, the model group had shortened colon length (P<0.01), increased DAI scores, spleen index, colon tissue pathology scores, and levels of TNF-α and IL-6 in serum (P<0.05, P<0.01), increased PI3K, phosphorylated Akt (p-Akt), and B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) expression in colon tissue (P<0.05, P<0.01), and decreased serum IL-10 levels and colon tissue Bcl-2 protein expression (P<0.01). Compared with the model group, the ZJW groups showed significant improvement in UC symptoms, relieved colon tissue pathological damage, downregulated levels of inflammatory cytokines TNF-α and IL-6 in serum (P<0.01), inhibited expression of PI3K, p-Akt, and Bax proteins in colon tissue (P<0.05, P<0.01), and increased serum IL-10 levels and colon tissue Bcl-2 protein expression (P<0.01), with the high-dose group showing the best effect. ConclusionZJW effectively alleviates DSS-induced UC, and its mechanism may be related to the inhibition of the PI3K/Akt signaling pathway and regulation of apoptosis-related protein expression.