1.Exploring Mechanism of Breast Cancer Prevention by Liuwei Dihuangwan, Erzhiwan and Zuoguiwan Based on Macrophage Phagocytosis
Lixiang ZHENG ; Yishan CHEN ; Wei QUAN ; Youbao ZHONG ; Zifeng GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):10-17
ObjectiveTo investigate the effects of three traditional Chinese medicine formulas for nourishing kidney Yin on the occurrence of breast cancer, including Zuoguiwan(ZGW), Liuwei Dihuangwan(LWDHW) and Erzhiwan(EZW), and to explore their preventive mechanisms from the phagocytic function of macrophages. MethodsTen-month-old post-breeding female mice were randomly divided into the blank group, soy isoflavone group(0.13 g·kg-1·d-1, mixed with feed), ZGW group(2.34 g·kg-1·d-1), LWDHW group(0.56 g·kg-1·d-1), and EZW group(4.68 g·kg-1·d-1). The mice were palpated every 3 d until the age of 18 months, and those with detected lumps were confirmed for tumor development through hematoxylin-eosin(HE) staining, and the incidence of breast cancer in mice was calculated. A total of 40 SPF-grade female SD rats were randomly divided into the blank serum group(physiological saline), ZGW drug-containing serum group(16.20 g·kg-1·d-1of ZGW), LWDHW drug-containing serum group(3.89 g·kg-1·d-1 of LWDHW) and EZW drug-containing serum group(32.40 g·kg-1·d-1 of EZW), each group was orally administered 3 times a day for 5 consecutive days. On the 5th day, blood was collected from the abdominal aorta 1 h after the last gavage to prepare drug-containing serum. The effect of drug-containing serum with different concentrations on the viability of RAW264.7 cells was assessed by cell counting kit-8(CCK-8). Taking 20% of drug-containing serum concentration as the research object, its effect on the expression levels of major histocompatibility complex-Ⅱ(MHC-Ⅱ), CD80 and CD86 on the surface of RAW264.7 cells was detected by immunofluorescence(IF), the phagocytic function of RAW264.7 cells was examined by flow cytometry, and the levels of interleukin-6(IL-6) and nitric oxide(NO) in RAW264.7 cells in the conditioned medium(CM) co culture system of 4T1 cells were detected by enzyme-linked immunosorbent assay(ELISA) and Griess assay. ResultsAfter prophylactic administration, the tumor incidence rates in the soy isoflavone group(4%), ZGW group(4%), LWDHW group(4%), and EZW group(6%) were lower than that in the blank group(8%). Compared with the blank serum group, the drug-containing serum of the three nourishing kidney Yin formulas could enhance the expression levels of MHC-Ⅱ, CD80 and CD86 on the surface of RAW264.7 cells, enhance phagocytic ability towards tumor cells, and reduce the content of IL-6 and increase the level of NO(P<0.05, P<0.01). ConclusionThe three nourishing kidney Yin formulas can reduce the incidence of tumor in mice, the mechanism may be related to activating RAW264.7 cells, increasing their phagocytosis to breast cancer cells, and regulating the secretion of IL-6 and NO.
2.Effect of Liuwei Dihuangwan on EMT and Expression of CSC Properties in 4T1 Cells by Regulating Myeloid-derived Suppressor Cells
Lixiang ZHENG ; Ling HUANG ; Huiwen GUO ; Biyao GONG ; Xiaoying REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):1-10
ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum (LDP) on epithelial-mesenchymal transition (EMT) and the expression of cancer stem cell (CSC) properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells (MDSCs). MethodsSPF-grade female SD rats were randomly divided into three groups, which were given 0.39, 1.94, 3.89 g·kg-1·d-1 suspension of Liuwei Dihuangwan for 7 days, respectively, to prepare low-, medium-, and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 (CCK-8) assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry (FC). The migration, invasion, and proliferation of 4T1 cells were detected by cell scratch assay, Transwell invasion assay, and plate colony-forming assay, respectively. Western blot (WB) was used to detect the protein expression of transforming growth factor-β (TGF-β), nuclear factor-κB (NF-κB), C-X-C motif chemokine ligand 2 (CXCL2), E-cadherin, and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence (IF). ResultsCompared with the normal group, LDP showed no significant inhibitory effect on the cell viability of 4T1 cells, but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs, as well as the expression of TGF-β (P<0.05, P<0.01). The migration, invasion, and proliferation of 4T1 cells were increased after co-culture with MDSCs (P<0.05, P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin and the proportion of CSC (CD44+CD24-) were elevated (P<0.05, P<0.01), while the expression of E-cadherin was decreased (P<0.05). After the intervention of MDSCs with LDP, followed by co-culture with 4T1 cells, the migration, invasion, and proliferation of 4T1 cells were obviously reduced (P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin were decreased (P<0.05, P<0.01), and the expression of E-cadherin was increased (P<0.05, P<0.01). There was no statistical difference in the proportion of CSC (CD44+CD24-) in 4T1 cells. However, the proportion of CSC (CD44+CD24-) was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention (P<0.05, P<0.01). ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β, NF-κB, and CXCL2, reverse EMT, and reduce the characteristic expression of CSC to inhibit the migration, invasion, and proliferation of 4T1 cells.
3.Effect of Liuwei Dihuangwan on EMT and Expression of CSC Properties in 4T1 Cells by Regulating Myeloid-derived Suppressor Cells
Lixiang ZHENG ; Ling HUANG ; Huiwen GUO ; Biyao GONG ; Xiaoying REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):1-10
ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum (LDP) on epithelial-mesenchymal transition (EMT) and the expression of cancer stem cell (CSC) properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells (MDSCs). MethodsSPF-grade female SD rats were randomly divided into three groups, which were given 0.39, 1.94, 3.89 g·kg-1·d-1 suspension of Liuwei Dihuangwan for 7 days, respectively, to prepare low-, medium-, and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 (CCK-8) assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry (FC). The migration, invasion, and proliferation of 4T1 cells were detected by cell scratch assay, Transwell invasion assay, and plate colony-forming assay, respectively. Western blot (WB) was used to detect the protein expression of transforming growth factor-β (TGF-β), nuclear factor-κB (NF-κB), C-X-C motif chemokine ligand 2 (CXCL2), E-cadherin, and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence (IF). ResultsCompared with the normal group, LDP showed no significant inhibitory effect on the cell viability of 4T1 cells, but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs, as well as the expression of TGF-β (P<0.05, P<0.01). The migration, invasion, and proliferation of 4T1 cells were increased after co-culture with MDSCs (P<0.05, P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin and the proportion of CSC (CD44+CD24-) were elevated (P<0.05, P<0.01), while the expression of E-cadherin was decreased (P<0.05). After the intervention of MDSCs with LDP, followed by co-culture with 4T1 cells, the migration, invasion, and proliferation of 4T1 cells were obviously reduced (P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin were decreased (P<0.05, P<0.01), and the expression of E-cadherin was increased (P<0.05, P<0.01). There was no statistical difference in the proportion of CSC (CD44+CD24-) in 4T1 cells. However, the proportion of CSC (CD44+CD24-) was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention (P<0.05, P<0.01). ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β, NF-κB, and CXCL2, reverse EMT, and reduce the characteristic expression of CSC to inhibit the migration, invasion, and proliferation of 4T1 cells.
4.Modern Thinking on the Indications of Daoyin (导引) based on the Theory of "Wei (痿) Diseases should be Treated by Daoyin"
Yan YU ; Xueyin CHEN ; Lihong YANG ; Shaonan LIU ; Yu CHEN ; Lixiang LIU ; Ping YUE ; Jinpeng WU ; Xinfeng GUO
Journal of Traditional Chinese Medicine 2024;65(11):1170-1173
It has been proposed by Basic Questions On Proper Therapies for Different Diseases Geographically (《素问·异法方宜论篇》) that "wei (痿) diseases should be treated by Daoyin (导引)". Furthermore, it is clarified that the indications of Daoyin are those conditions related to spleen and dampness caused by dampness pathogen, excessive food intake and less exercise, and mainly manifested as heavy limbs, fatigue and flaccidity, which is similar to the metabolic imbalance in the early stage of glucose or lipid metabolism disorder in modern medicine. Based on modern clinical and basic research evidence, Daoyin can inhibit the response of inflammation, alleviate oxidative stress, regulate intestinal microbiota, and modulate gene expression to improve metabolic abnormalities, and this will provide ideas for researches on the indications of Daoyin.
5.Liuwei Dihuang Erzhiwan Combination Regulate Myeloid-derived Suppressor Cells to Inhibit Breast Cancer Lung Metastasis
Lixiang ZHENG ; Zifeng GUO ; Huiwen GUO ; Xiaomin WANG ; Chuanming XU ; Yuliang HU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(20):37-45
ObjectiveTo investigate the mechanism by which Liuwei Dihuang Erzhiwan combination inhibit the lung metastasis of spontaneous breast cancer in mice by regulating the recruitment of myeloid-derived suppressor cells (MDSCs). MethodThree hundred and eighty SPF-grade 10-month-old female breeders of Kunming mouse were palpated at the mammary gland site once every 3 days. Mice that have not had a lump touched after being raised for 6 months are used as control group. After tumor development, the mice were randomized into model, positive control (paclitaxel, intraperitoneal injection at 0.01 g·kg-1 every other day for 22 d), Liuwei Dihuangwan (0.65 g·kg-1·d-1 by gavage), Erzhiwan (5.41 g·kg-1·d-1 by gavage), and Liuwei Dihuang Erzhiwan combination (6.05 g·kg-1·d-1 by gavage) groups. The mice were euthanised when the tumor reached a diameter of about 15 mm, and the tumor and lung tissues were collected. The survival time, tumor mass, and lung metastasis rate of tumor-bearing mice were recorded. Hematoxylin-eosin (HE) staining was used to observe the histopathological and morphological changes of mouse tumor and lung tissues. Immunofluorescence (IF) was used to detect the distribution of MDSCs in tissues of mice in each group by double-staining of MDSCs cells with lymphocyte antigen 6 complex site G6D (Ly6G) and CD11 antigen-like family member B (CD11b). Western blot was employed to determine the protein levels of matrix metalloproteinase-9 (MMP-9), transforming growth factor-β (TGF-β), zinc finger transcription factor 1 (Snail1), and E-cadherin in the tumor tissue and CC motif chemokine 9 (CCL9) and CC motif chemokine receptor 1 (CCR1) in the lung tissue. ResultDuring the modelling period, the paclitaxel group and Chinese medicine intervention groups had longer median number of days of survival and lower tumor weight, lung metastasis rate, and lung nodule than the model group (P<0.05, P<0.01). HE staining showed an increase in tumor cell necrosis in the paclitaxel group and the Liuwei Dihuang Erzhiwan combination group. The paclitaxel group and Chinese medicine intervention groups had lower fluorescence intensity of MDSCs in the tumor tissue than the model group (P<0.05, P<0.01). Compared with the normal control group, the model group showed increased fluorescence intensity of MDSCs in the metastatic lung tissue (P<0.01), which, however, was decreased in the paclitaxel group and Chinese medicine intervention groups (P<0.01). The model group showed higher protein levels of MMP-9, TGF-β, and Snail1 and lower protein level of E-cadherin in the tumor tissue than in the normal control group (P<0.01). Compared with model group, paclitaxel and Chinese medicine interventions downregulated the protein levels of MMP-9, TGF-β, and Snail1 (P<0.05, P<0.01) and upregulated the protein level of E-cadherin in the tumor tissue (P<0.01). Moreover, the Liuwei Dihuang Erzhiwan combination group had lower protein levels of TGF-β and Snail1 than the Liuwei Dihuangwan group and Erzhiwan group (P<0.05). In the metastatic lung tissue, the expression of CCL9 and CCR1 was higher in the model group than in the normal control group, paclitaxel group, and Chinese medicine intervention groups (P<0.05, P<0.01). ConclusionLiuwei Dihuang Erzhiwan combination inhibit tumor growth, prolong survival time, and reduce the occurrence of lung metastasis in the mouse model of spontaneous breast cancer by reducing the recruitment of MDSCs in the tumor and lung tissues and modulating the phenotypes of epithelial-mesenchymal transition (EMT)-related molecules and the expression of CCL9/CCR1.
6.Research progress of heat shock protein 90 in breast cancer
Maolin GUO ; Chao ZHENG ; Lixiang YU ; Liyuan LIU ; Linfeng ZHAO ; Zhigang YU
Chinese Journal of Endocrine Surgery 2023;17(2):249-252
Breast cancer has become the most common malignant tumor in the world. Heat shock protein 90 (HSP90) is a kind of molecular chaperone which can promote protein folding and maintain protein stability. HSP90 includes HSP90α, HSP90β, GRP94 and TRAP1 subtypes. Previous studies have found that the level of HSP90 is significantly increased in malignant tumors such as breast cancer, and is closely related to the occurrence and development of tumors. Meanwhile, the research on inhibitors targeting HSP90 has also attracted much attention. In this paper, we reviewed the expression of four HSP90 subtypes in breast cancer and their relationship with the clinicopathologic feature and prognosis of patients, discussed the research progress of specific inhibitors of HSP90 subtypes in breast cancer, and analyzed the application prospect of HSP90 as biomarkers for breast cancer prognosis monitoring and therapeutic targets.
7. Morphologic variations of the fibular malleolar groove and fibrocartilaginous ridge injury on MRI with and without re-current peroneal tendon dislocation
En DENG ; Lixiang GAO ; Qinwei GUO
Chinese Journal of Orthopaedics 2019;39(9):562-566
Objective:
To compare the morphological variation of the fibular malleolar groove and fibrocartilaginous ridge injury in those with and without peroneal tendon dislocation using MRI.
Methods:
Data of 45 patients (40 males, 5 females; average age 23.4±7.7 years, 11-44 years) with recurrent peroneal tendon dislocation treated from April 2010 to July 2017 and 45 patients (33 males, 12 females; average age 41.6±11.8 years, 17-67 years) without dislocation treated from June 2015 to Septem-ber 2017 were retrospectively analyzed. The morphological variation of the fibular malleolar groove and fibrocartilaginous ridge in-jury were compared between the two groups according to Rosenberg’s classification (convex, the posterior cortex of the fibula bulg-es outward; concave, the posterior fibular surface has a shallow-to-deep depression; flat, neither concavity nor convexity is found).
Results:
In group of patients with recurrent peroneal tendon dislocation: concave, 10 patients (22.2%, 10/45); flat, 27 patients (60.0%, 27/45); convex, 8 patients (17.8%, 8/45); fibrocartilaginous ridge injury, 44 patients (97.8%, 44/45). In group of patients without peroneal tendon dislocation: concave, 16 patients (35.6%, 16/45); flat, 18 patients (40.0%, 18/45); convex,11 patients (24.4%, 11/45); fibrocartilaginous ridge injury, 13 patients (28.9%, 13/45).
Conclusion
According the results, significant differ-ence has not been found in morphologic variations of the fibular malleolar groove between patients with and without recurrent pero-neal tendon dislocation. Fibrocartilaginous ridge injury is common in MRI of patients with recurrent peroneal tendon dislocation.
8.Correlation analysis between thrombelastography and conventional coagulation indexes in breast cancer patients
Zilan LYU ; Binlin WANG ; Bianqin GUO ; Yang ZHANG ; Lixiang WU
International Journal of Laboratory Medicine 2019;40(2):218-221
Objective To analyze the correlation between thrombelastography (TEG) and conventional coagulation indexes in breast cancer patients, and to compare the differences between the two methods in the detection of coagulation function in breast cancer patients.Methods Retrospectively analyzed the clinical data of180patients with breast cancer who were performed TEG, coagulation and blood test in the same day in our hospital from November 2016to May 2017.Linear correlation and regression analysis were performed among the parameters.The differences of positive rates of TEG parameters and conventional coagulation indexes were compared.Results The R value of TEG parameters of breast cancer patients was positively correlated with K and APTT, and negatively correlated withα-Angle, MA, CI, DD, FDPs.K was positively correlated with APTT and TT, and negatively correlated withα-Angle, MA, FIB, DD, FDPs and PLT.α-Angle was positively correlated with MA, CI, FIB, DD, FDPs and PLT.MA was positively correlated with CI, FIB, DD, FDPs and PLT.CI was positively correlated with FIB, DD, FDPs and PLT.α-Angle, MA and CI were all negatively correlated with APTT and TT, the difference were statistically significant (P<0.05).A linear regression equation of TEG parameters and coagulation indexes was obtained.There was no significant difference between TEG detection positive rate and conventional coagulation test (P>0.05).Conclusion There are significant correlation between the TEG parameters and routine coagulation or platelet, and the results are consistent.
9.Correlation analysis between thrombelastogram and coagulation test and platelet count in patients with malignant tumor
Zilan LV ; Binlin WANG ; Yang ZHANG ; Bianqin GUO ; Bo LONG ; Xiaohua WANG ; Lixiang WU
International Journal of Laboratory Medicine 2018;39(4):443-446,449
Objective To analyze the correlation between thrombelastography(TEG)and coagulation, platelet count(PLT)in patients with malignant tumor.Methods Retrospectively analyzed the clinical data of 241 cases with tumor who were performed TEG,coagulation and blood test in the same day in Chongqing Cancer Institute from November 2016 to March 2017.Linear correlation and regression were carried out to an-alyze relationship among the parameters.The number of patients with positive blood clotting,PLT and TEG parameters were counted,and the χ2test was used to compare the difference between them.Using Mann-Whit-ney U test to compare the differences between multiple parameters of liver cancer,breast cancer,and pancreatic cancer.Results The R value of TEG parameters in patients with malignant tumor was positively correlated with APTT,negatively correlated with TT,DD and FDPs.K was positively correlated with APTT and TT, and negatively correlated with FIB and PLT.The relationship between α and FIB,PLT were positive,between APTT and TT were negative.MA was positively correlated with FIB and PLT,negatively correlated with TT, CI was positively correlated with FIB and PLT,and negatively correlated with APTT(P<0.05).The correla-tion between FIB,PLT and MA was the highest.And the linear regression equation of TEG parameters and coagulation indexes was obtained.The positive rate of TEG was lower than that of coagulation(P<0.05). Same certain differences of TEG and coagulation parameters were existed in liver cancer,breast cancer and pancreatic cancer patients.Conclusion TEG is significantly associated with PLT and conventional coagulation test,and the results of TEG and conventional coagulation test are consistent to a certain degree,but the overall agreement is generally not interchangeable.TEG might be play a complementary role with coagulation tests and platelet counts.And the TEG of different cancer types is not exactly the same as the coagulation parameters.
10. Clinical analysis of video-assisted thoracoscopic surgery in 65 patients with early non-small cell lung cancer
Lixiang HE ; Shengcong GUO ; Botao XU
Chinese Journal of Primary Medicine and Pharmacy 2018;25(9):1145-1148
Objective:
To study the clinical effect of video-assisted thoracoscopic surgery in the treatment of early non-small cell lung cancer(NSCLC).
Methods:
130 cases with early NSCLC were selected.According to random number table, the patients were divided into the observation group and the control group, 65 cases in each group.The observation group was treated with video-assisted thoracoscopic surgery, while the control group was treated with thoracotomy.The perioperative period of the two groups was compared, 24h before operation and 24h after operation, the changes of T lymphocyte subsets(CD3+ , CD4+ , CD4+ /CD8+ ) were measured, and the postoperative complications were recorded.The patients were followed up for one year, the survival rate and recurrence rate were recorded.
Results:
There was no statistically significant difference in the number of lymph node dissection between the two groups(

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