1.The Changes of HIF-1α and VEGF-A in Myocardial Tissue of Rats with Arrhyth-mias
Yuan ZHANG ; Zhipeng CAO ; Ruiming MAO ; Zhongbo DU ; Li MI ; Xinyi LUO ; Meihui TIAN ; Baoli ZHU
Journal of Forensic Medicine 2017;33(3):225-231
Objective T o observe the expression changes of hypoxia inducible factor-1α (H IF-1α) and vascular endothelial grow th factor-A (V E G F-A ) in rats w ith arrhythm ias, and to explore the differences of the expression pattern in the tw o indicators of acute m yocardial ischem ia caused by arrhythm ias and coronary insufficiency. Methods T he arrhythm ia w as induced by C aC l2, and the expression changes of H IF-1α and V E G F-A w ere detected by im m unohistochem istry, W estern blotting and real-tim e PC R w ithin 6 h after the arrhythm ia in rats. Results T he expression of H IF-1α and V E G F-A show ed diffuse in the m yocardial tissue of rats died from arrhythm ias. B oth of them increased in the early arrhythm ia, then decreased. E xtensive m yocardial ischem ia happened at the beginning of arrhythm ia occurrence and its range didn't expand w ith tim e. Conclusion T he expressions of H IF-1α and V E G F-A in m yocardium of the rats w ith arrhythm ia can provide evidence for the differential diagnosis of acute m yocardial is-chem ia caused by fatal arrhythm ia and coronary insufficiency.
2.Biochemical Indicators of Anaphylactic Shock and the Application in Forensic Medicine
Li MI ; Jie CHEN ; Weimin GAO ; Zhongbo DU ; Zhipeng CAO ; Yuan ZHANG ; Baoli ZHU
Journal of Forensic Medicine 2014;(2):117-121
Fatal anaphylactic shock is common in forensic practice. However, it is difficult to diagnose for lacking specific pathological and morphologic changes in forensic autopsy. The application of some biochemical indicators is of great significance. This paper reviews the biological characteristics of some biochemical indicators and detection methods. The forensic application, problems and prospects of these indicators are also introduced in details. The stable biochemical indicators, IgE, tryptase and chymase, show great potential and advantages in the identification of fatal anaphylactic shock in forensic medicine.
3.Changes of Serum IgE and Tryptase in Anaphylactic Shock Rats
Li MI ; Weimin GAO ; Zhongbo DU ; Zhipeng CAO ; Yuan ZHANG ; Baoli ZHU
Journal of Forensic Medicine 2015;(3):181-184
Objective To explore the changes of serum IgE and tryptase caused by anaphylactic shock rats and discuss the relation to PMI and preservative environm ent of corpse and specim en. Methods Rats were used for establishing anaphylactic shock m odels and random ly divided into room tem perature group, refrigeration group, frozen group, manual hem olysis group, specim en preservation group. And the control group was also established. The blood sam ples were collected after rats were sacrificed. The de-gree of hem olysis was graded according to the color of the upper layer of the serum . The mass concen-tration of IgE and tryptase in each group was detected by ELISA. Results The levels of serum IgE and tryptase in anaphylactic shock dead rats were higher than that of the control group. Room tem perature and frozen m ade obviously differences on the levels of serum IgE and tryptase with various PMI. The levels of serum IgE and tryptase in refrigeration group show ed relatively stable. The levels of serum tryptase and IgE were elevated with differently increasing hem olysis. The levels of serum IgE and tryptase show ed no obvious changes during the specim en kept under different tem perature conditions for 25 days. Conclusion Serum IgE and tryptase obviously increased in anaphylactic shock rats. H ow ever, the levels were influenced by PMI and environm ental tem perature, especially under the conditions of room tem perature and frozen.
4.Effect of Yanghe decoction containing serum on expression of iNOS, COX-2 and NF-κB in breast cancer MCF-7 cells
Liying LIU ; Jianwei DOU ; Gaobiao LI ; Shuo YANG ; Zhongbo ZHU ; Qian HUANG
International Journal of Traditional Chinese Medicine 2019;41(2):173-176
Objective To observe the changes of iNOS, COX-2 and NF-κB in breast cancer MCF-7 cells treated with Yanghe decoction containing serum. Methods The MCF-7 human breast cancer cells were divided into blank group, low dose group, middle dose group and high dose group (4.5, 9, 18 g/kg, respectively). Real-time RT-PCR was used to detect the mRNA expression of iNOS and COX-2 at 24, 48, 72 and 96 hours, and Western-blot was used to detect the expression of NF-κB protein. Results Compared with the blank group, the expression of iNOS and COX-2 mRNA in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours(P<0.05), the the expression of NF-κB protein in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours (P<0.05). So it reduced with the increase of drug concentration and time. There was no significant difference in 72 hours after intervention. Conclusions The Yanghe decoction could reduce the expression of NF-κB, and then reducing the related inflammatory factors COX-2 and iNOS.
5. Effect of Yanghe decoction on the expression of Smad3 and NF-κB in breast cancer MDA-MB-231 cells
Jianwei DOU ; Zhongbo ZHU ; Kangle LI ; Shuo YANG ; Qian HUANG
International Journal of Traditional Chinese Medicine 2019;41(10):1087-1090
Objective:
To investigate the effect of
6.Mechanism of Banxia Xiexintang in Prevention and Treatment of Gastric Cancer: A Review
Yanmei ZHANG ; Jingjing WEI ; Zhongbo ZHU ; Senyu LIU ; Fan YANG ; Xiping LIU ; Peiqing LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):65-72
Gastric cancer (GC) is one of the common malignant tumors, and the incidence and mortality of GC in China rank first in the world. At present, the pathogenesis of GC has not been fully clarified. Although surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy have achieved good results in the treatment of GC, there are still many complications, decreased sensitivity, and severe side effects. Banxia Xiexintang, derived from Treatise on Cold Damage and Miscellaneous Diseases(《伤寒杂病论》), has been clinically used for more than 2000 years with the effects of combining cold and warm drugs, dissipating mass, and relieving stuffiness, and is a classic prescription for treating digestive tract diseases in later generations. Through clinical observation and experimental research, it is found that Banxia Xiexintang and its single drugs have good effect in preventing and treating GC. Chinese medicine has multi-component and multi-target characteristics and can treat GC through various mechanisms. Therefore, it is necessary to carry out systematic and in-depth research from the aspects of molecular biology and network pharmacology, and comprehensively reveal the mechanism of Banxia Xiexintang in preventing and treating GC. At present, the mechanism of Banxia Xiexintang in treating GC mainly focuses on inducing apoptosis of GC cells, inhibiting proliferation, migration, and invasion of GC cells, protecting peritoneal mesothelial cells, inhibiting peritoneal metastasis of GC cells, regulating GC microenvironment, and inhibiting the malignant transformation of bone marrow mesenchymal stem cells (BMSCs). This research group is committed to the prevention and treatment of GC with Banxia Xiexintang, aiming to comprehensively reveal the mechanism of action and the pharmacodynamic material basis of Banxia Xiexintang in the prevention and treatment of GC, and provide an important scientific basis for further clinical application of Banxia Xiexintang. After searching CNKI, PubMed, Wanfang Data, VIP, and other databases, this paper summarized Banxia Xiexintang in the treatment of GC from the aspects of prescription basis, material basis, network pharmacology, clinical and experimental studies, etc., so as to provide references for further research on pharmacological effect of Banxia Xiexintang and its application in the clinical treatment of GC.
7.Mechanism of Shenqi Yiliu Prescription Combined with Cisplatin on H22 Liver Cancer-bearing Mice Based on NLRP3/Caspase-1/GSDMD Pyroptosis Pathway
Mengying YANG ; Yongqiang DUAN ; Yuxin JIA ; Min BAI ; Zhongbo ZHU ; Yarong LI ; Lan MA ; Mingyu ZHANG ; Xin FENG ; Lanlan HE ; Yuping YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):114-122
ObjectiveTo explore the anti-tumor effect and mechanism of Shenqi Yiliu prescription in the intervention of pyroptosis. MethodTen male BALB/c mice were randomly selected and assigned to the blank group. The remaining 40 mice underwent the induction of the liver cancer xenograft model. After 5 days of modeling, 40 surviving mice were randomly divided into model group, cisplatin group [2.5×10-3 g·kg-1·(3 d)-1], Shenqi Yiliu prescription group (27 g·kg-1·d-1), and a combination group (Shenqi Yiliu prescription group + cisplatin). The mice in the blank group and the model group were treated with an equal volume of normal saline for 10 days. The general conditions of mice in each group were observed. After the intervention, the tumor weight of the mice was weighed and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in tumor tissues. The levels of mouse liver function indicators, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. The TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay was used to detect DNA damage in mouse tumor tissue cells. Immunohistochemistry (IHC), immunofluorescence (IF), and Western blot were used to detect the protein expression levels of NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate-specific protease-1 (Caspase-1), and gasdermin D (GSDMD) in tumor tissues. The levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in tumor tissues were detected by enzyme-linked immunosorbent assay (ELISA). ResultCompared with the mice in the blank group, those in the model group were in a poor mental state, sleepy, and lazy, and their fur color was dull, with increased levels of serum ALT and AST in liver function tests (P<0.01). Compared with the model group, the groups with drug intervention showed improved mental state, inhibited tumor growth to varying degrees, and decreased tumor weight, and the tumor inhibition rate in the combination group was the highest (P<0.01). HE staining showed that the pathological and morphological lesions of the tumor tissues in the model group were significant, while those in all groups with drug intervention were improved to a certain extent. The karyolysis and nuclear rupture in the Shenqi Yiliu prescription group and the combination group were more significant. In the liver function test, the serum ALT and AST levels of mice in the Shenqi Yiliu prescription group and the combination group decreased (P<0.01), and the inflammatory factors IL-1β and IL-18 in each group with drug intervention decreased (P<0.05, P<0.01). Among them, the declining trend of IL-1β and IL-18 in the Shenqi Yiliu prescription group was the most significant (P<0.01). TUNEL staining showed that the positive TUNEL staining in each group with drug intervention decreased after intervention (P<0.05, P<0.01), especially the cisplatin group and Shenqi Yiliu prescription group (P<0.01). Western blot, IHC, and IF found that the protein expression levels of NLRP3, Caspase-1, and GSDMD in each group with drug intervention decreased (P<0.05, P<0.01). Compared with the mice in the cisplatin group, those in the Shenqi Yiliu prescription group and the combination group had better mental state and regular tumor morphology, and the tumor weight of the mice in the combination group decreased (P<0.05). The levels of ALT and AST in the Shenqi Yiliu prescription group decreased (P<0.05), and the levels of IL-1β and IL-18 in the Shenqi Yiliu prescription group and the combination group decreased (P<0.05, P<0.01), especially in the combination group (P<0.01). The results of IHC showed that the expression of GSDMD protein in the tumor tissues of mice in the combination group was reduced (P<0.01). IF detection showed that the expression of NLRP3 in the tumor tissues of the Shenqi Yiliu prescription group was reduced (P<0.01). The results of Western blot showed that the expression level of NLRP3 protein in the Shenqi Yiliu prescription group and the combination group decreased (P<0.01), and the expression level of Caspase-1 protein in the combination group decreased (P<0.01). The decrease in GSDMD protein expression was not significant, and the difference was not statistically significant. ConclusionShenqi Yiliu prescription combined with cisplatin has an obvious anti-tumor effect, which may be achieved by down-regulating the NLRP3/Caspase-1/GSDMD inflammatory pyroptosis pathway to inhibit cell pyroptosis, and relieve the inflammatory response in mice with liver cancer.
8.Effect of Banxia Xiexintang-containing Intestinal Absorption Solution on Migration and Invasion of PMN-MDSCs in Gastric Cancer Microenvironment
Jingjing WEI ; Zhongbo ZHU ; Xiping LIU ; Peiqing LI ; Qiming CHEN ; Lirong DAI ; Lijuan SHI ; Haijing DUAN ; Qingmiao WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):48-57
ObjectiveTo observe the effect of Banxia Xiexintang containing intestinal absorption solution (BXCIAS) on migration and invasion of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in gastric cancer microenvironment. MethodThe complex solution (containing 0.63 g·mL-1 crude drug) was prepared. Gastric cancer cells were subjected to non-contact co-culture with PMN-MDSCs in Transwell chamber to create gastric cancer microenvironment. Cell counting kit-8 (CCK-8) assay was used to screen the optimal intervention concentration and time of BXCIAS on PMN-MDSCs for subsequent experiment. The blank group, model group, FAK inhibitor group, and BXCIAS groups (26%, 18%, and 10%) were designed. Scratch assay and Transwell assay were employed to detect the migration and invasion ability of PMN-MDSCs, and enzyme-linked immunosorbent assay (ELISA) to measure the expression of vascular endothelial cell growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in tumor microenvironment. The expression levels of PMN-MDSCs pathway-related proteins FAK, phosphorylated (p)-FAK, protein tyrosine kinase (Src), and p-Src were detected by Western blot. ResultThe inhibition rates of PMN-MDSCs by 5%, 50%, 75%, and 100% BXCIAS at 48 h were higher than those at 24 h (P<0.05, P<0.01). The inhibition rate of PMN-MDSCs by 50% BXCIAS at 72 h was lower than that at 48 h (P<0.01), and the inhibition rates by 5% and 100% BXCIAS at 72 h were higher than those at 48 h (P<0.05, P<0.01). There was no significant difference in the inhibition rate by other concentration levels at 48 h. The half-maximal inhibitory concentration (IC50) at 48 h was 18.09%, indicating that 18% BXCIAS and 48 h were the optimal concentration and time, respectively. The migration distance of PMN-MDSCs was large (P<0.01), and the number of migrating and invading cells increased (P<0.01) in the mode group compared with those in the blank group. Compared with model group, FAK inhibitor and BXCIAS at different concentration decreased the migration distance of PMN-MDSCs (P<0.01), and the number of migrating and invading cells (P<0.01), especially the 26% BXCIAS (P<0.01). The expression of PMN-MDSCs pathway-related proteins FAK, p-FAK, Src and p-Src (P<0.01) and the expression of VEGF and MMP-9 (P<0.01) were higher in the model group than in the blank group. Compared with model group, FAK inhibitor and BXCIAS (26%, 18%, 10%) decreased the expression of FAK, p-FAK, and Src (P<0.01), and FAK inhibitor and 18% BXCIAS reduced the expression of p-Src (P<0.01), and the expression of VEGF and MMP-9 (P<0.01). ConclusionBXCIAS can inhibit the migration and invasion of PMN-MDSCs by down-regulating the expression of FAK, p-FAK, Src, and p-Src proteins in the FAK signaling pathway of PMN-MDSCs in gastric cancer microenvironment.
9.Effect of Banxia Xiexintang-containing Intestinal Absorption Solution on PMN-MDSCs Apoptosis in Gastric Cancer Microenvironment
Jingjing WEI ; Zhongbo ZHU ; Xiping LIU ; Peiqing LI ; Qiming CHEN ; Lirong DAI ; Lijuan SHI ; Haijing DUAN ; Qingmiao WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):58-64
ObjectiveTo observe the effect of Banxia Xiexintang (BXT)-containing intestinal absorption solution on the apoptosis of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in gastric cancer microenvironment. MethodBXT-containing intestinal absorption solution was prepared, and gastric cancer cells and PMN-MDSCs were non-contact co-cultured in Transwell chamber to establish gastric cancer microenvironment. Cell counting kit-8 (CCK-8) assay was used to screen the optimal intervention concentration and time of 0-100% BXT-containing intestinal absorption solution prepared by 0.63 g·mL-1 reconstitution solution. Cells were classified into blank group, model group, oxaliplatin group (10 mg·L-1), and BXT (26%, 18%, 10% BXT-containing intestinal absorption solution) group, and the apoptosis of PMN-MDSCs was detected by flow cytometry. The expression of apoptosis-related B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteine-aspartic acid protease-3 (Caspase-3) in PMN-MDSCs was detected by Western blot. ResultAfter treatment for 24 h and 48 h, the PMN-MDSCs-inhibiting rate was increased by 5%, 50%, 75%, and 100% BXT-containing intestinal absorption solution compared with that in the blank group (P<0.05, P<0.01). At 72 h, the PMN-MDSCs-inhibiting rate by 50% BXT-containing intestinal absorption solution was lower than that at 48 h (P<0.01), and the PMN-MDSCs-inhibiting rate by 5%, 75%, and 100% BXT-containing intestinal absorption solution showed no significant difference from that at 48 h. Moreover, the half-maximal inhibitory concentration (IC50) at 48 h was 18.40%. Thus, 18% BXT-containing intestinal absorption solution and 48 h were the optimal intervention concentration and time. The survival rate of PMN-MDSCs in model group was higher than that in the blank group (P<0.05), and the apoptosis rate was lower than that in the blank group (P<0.05). Compared with model group, BXT containing intestinal absorption solution lowered the survival rate and raised apoptosis rate of PMN-MDSCs (P<0.05), particularly the 26% BXT-containing intestinal absorption solution (P<0.05). The expression of Bax and Caspase-3 in PMN-MDSCs was lower in the model group than in the blank group (P<0.05), and the expression of Bcl-2 was higher in the model group than in the blank group (P<0.05). The expression of Caspase-3 in PMN-MDSCs increased (P<0.05) and the expression of Bcl-2 decreased (P<0.05) in oxaliplatin group and BXT group compared with those in the model group. The expression of Bax rose in oxaliplatin group and BXT group (10% BXT-containing intestinal absorption solution) (P<0.05). ConclusionBXT can induce the apoptosis of PMN-MDSCs by regulating the expression of apoptosis-related proteins Bax, Caspase-3, and Bcl-2 in gastric cancer microenvironment.