Qizhu anticancer prescription enhances immunosurveillance of liver cancer cells by regulating p21-dependent secretory phenotypes.

ETHNOPHARMACOLOGICAL RELEVANCE: Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide, largely due to the limitations of available therapeutic strategies. The traditional Chinese medicine Qizhu Anticancer Prescription (QZACP) can improve the quality of life and prolong the survival time of patients with HCC. However, the precise mechanisms underlying the anti-cancer properties of QZACP remain unclear. PURPOSE: This study examined the anti-hepatocarcinogenic properties of QZACP, with a specific focus on its influence on the p21-activated secretory phenotype (PASP)-mediated immune surveillance, to elucidate the underlying molecular pathways involved in HCC. MATERIALS AND METHODS: Cell proliferation was measured using the Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine, and clonogenic assays. The cell cycle was evaluated using flow cytometry, and senescence was identified by staining with senescence-associated beta-galactosidase (SA-ß-gal). A primary liver cancer model produced by diethylnitrosamine was established in C57 BL/6 mice to assess the tumor-inhibitory effect of QZACP. The liver's pathological characteristics were examined using hematoxylin and eosin staining. PASP screening was performed using GeneCards, DisGeNet, Online Mendelian Inheritance in Man, and The Cancer Genome Atlas databases. Western blot analysis, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, and Transwell migration assays were performed. RESULTS: Serum containing QZACP enhanced p21 expression, triggered cell cycle arrest, accelerated cell senescence, and suppressed cell proliferation in Huh7 and MHCC-97H liver cancer cells. QZACP reduced the quantity and dimensions of liver tumor nodules and enhanced p21 protein expression, SA-ß-Gal staining in tumor lesions, and cytotoxic CD8+ T cell infiltration. Bioinformatic analyses indicated that PASP factors, including hepatocyte growth factor, decorin (DCN), dermatopontin, C-X-C motif chemokine ligand 14 (CXCL14), and Wnt family member 2 (WNT2), play an important role in the development of HCC. In addition, these factors are associated with the presence of natural killer cells and CD8+ T cells within tumors. Western blotting and ELISA confirmed that QZACP increased DCN, CXCL14, and WNT2 levels in tumor tissues and peripheral blood. CONCLUSIONS: QZACP's suppression of HCC progression may involve cell senescence mediated via p21 upregulation, DCN, CXCL14, and WNT2 secretion, and reversal of the immunosuppressive microenvironment. This study provides insights that can be used in the development of new treatment strategies for HCC.

Investigating the molecular mechanism of Qizhu anticancer prescription in inhibiting hepatocellular carcinoma based on high-resolution mass spectrometry and network pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Of all primary liver cancer cases, hepatocellular carcinoma (HCC) accounts for about 90%. Most patients with HCC receive a diagnosis in the medium-to-late stages or with chronic liver disease, have lost the opportunity for radical treatment, such as surgical resection, and their 5-year survival rate is low. Qizhu Anticancer Prescription (QZACP) is an empirical formula composed of traditional Chinese herbs that can clinically relieve HCC symptoms, inhibit the progression of HCC, reduce recurrence rate, and prolong survival; however, its exact mode of action remains unknown. AIM OF THE STUDY: This study's purpose was to investigate the mode of action of QZACP in the prevention and treatment of HCC. MATERIALS AND METHODS: Initially, drug components in the QZACP decoction were analyzed using high-resolution mass spectrometry. A subcutaneous tumor xenograft model in nude mice was constructed to further analyze the active components of QZACP that had entered tumor tissues through oral administration. Potential targets of QZACP in the prevention and treatment of HCC were identified and then confirmed in vivo via network pharmacology and molecular docking. In addition, regulatory effects of QZACP on HCC cell proliferation and the cell cycle were detected using a CCK-8 assay and flow cytometry. RESULTS: High-resolution mass spectrometry revealed that the QZACP decoction contained deacetyl asperulosidic acid methyl ester (DAAME), paeoniflorin, calycosin-7-glucoside, liquiritin, glycyrrhizic acid, astragaloside IV, saikosaponin A, curdione, and atractylenolide II. In nude mice, QZACP could effectively inhibit the growth of subcutaneous tumors, where DAAME, paeoniflorin, liquiritin, and glycyrrhizic acid could enter liver cancer tissues after oral administration. Among these, DAAME was the most highly expressed in HCC tissues and may be an important active component of QZACP for inhibiting HCC. Utilizing network pharmacology, the targets of action of these four drug components were identified. After verification using western blotting, STAT3, VEGFA, JUN, FGF2, BCL2L1, AR, TERT, MMP7, MMP1, ABCB1, CA9, and ESR2 were identified as targets of QZACP inhibition in HCC. In vitro experiments revealed that QZACP inhibited the proliferation of HCC cells while inducing G0/G1 phase cell cycle arrest. In vivo experiments demonstrated that DAAME significantly inhibited HCC growth. After intersection of the 24 DAAME targets predicted using network pharmacology with the 435 HCC disease targets, only CA9 was identified as a DAAME-HCC crossover target. Molecular docking results revealed that the binding site of DAAME and CA9 had good stereo-complementarity with a docking score of -8.1 kcal/mol. Western blotting and immunohistochemical results also confirmed that DAAME significantly decreased CA9 protein expression in HCC. CONCLUSIONS: QZACP inhibits HCC by reducing the expression of STAT3, VEGFA, JUN, FGF2, BCL2L1, AR, TERT, MMP7, MMP1, ABCB1, CA9, and ESR2. DAAME may be an important active component of QZACP for the prevention and treatment of HCC, inhibiting it by targeting the expression of CA9.

Mechanism of Non-alcoholic Steatohepatitis Improved by Qizhu Prescription Based on AMPK Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effect and mechanism of Qizhu prescription on liver lipid anabolism and oxidative stress in mice with non-alcoholic steatohepatitis （NASH） based on adenylate activated protein kinase （AMPK） signaling pathway. MethodA total of 60 male C57BL/6J mice were randomly divided into a normal group （n = 10） and a modeling group （n = 50）. The modeling group was fed by high-fat and high-cholesterol diet for 16 weeks to establish the NASH mice model and was randomly divided into model group， low-， medium， and high-dose groups of Qizhu prescription， and Yishanfu group， with 10 mice in each group. Qizhu prescription was administered intragastrically once a day at a dose of 4.75， 9.50， and 19.00 g·kg-1 in each group and 228 mg·kg-1 in Yishanfu group. The normal group and model group were given equal volumes of pure water for eight weeks. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total cholesterol （TC）， triglyceride （TG）， and glucose （GLU） levels were detected. The pathological changes of liver tissue were observed by hematoxylin-eosin （HE） and oil red O staining. Serum levels of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， free fatty acids （FFA）， reduced glutathione （GSH）， malondialdehyde （MDA）， catalase （CAT）， and superoxide dismutase （SOD） were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression levels of acetyl-CoA carboxylase （ACC）， carnitine palmitoyl transferase 1A（CPT1A）， and mitochondrial uncoupling protein 2 （UCP2） were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Protein expression levels of AMPK， p-AMPK， ACC， CPT1A， and UCP2 in liver tissue were detected by Western blot. ResultCompared with the normal group， the liver steatosis of the model group was obvious， with multiple inflammatory clusters and large amounts of intracellular lipid deposition. The activity of serum AST， ALT， as well as levels of IL-6， IL-1β， TNF-α， FFA， and MDA were significantly increased， the activity of CAT and SOD was significantly decreased， and the mRNA and protein expressions of ACC were significantly increased. The mRNA and protein expressions of CPT1 and UCP2 were significantly decreased， and the protein expression of p-AMPK was significantly decreased （P<0.01）. Compared with the model group， the degree of liver steatosis in the Qizhu prescription and Yishanfu groups was reduced， the activity of AST and ALT， as well as the levels of IL-6， IL-1β， TNF-α， FFA， and MDA was significantly decreased， and the activity of CAT and SOD was significantly increased （P<0.01）. The mRNA and protein expressions of ACC in liver tissue of mice in medium- and high-dose groups of Qizhu prescription were significantly decreased， while the mRNA and protein expressions of CPT1A and UCP2， as well as p-AMPK protein were significantly increased （P<0.01）. ConclusionQizhu prescription can improve liver lipid metabolism， reduce oxidative stress， and promote liver cell repair in NASH mice by activating the AMPK signaling pathway.

Mechanism of Qizhu Kang'ai Prescription for Inhibiting Proliferation of Hepatocellular Carcinoma by Regulating Tumor Metabolic Reprogramming via PCK1/Akt/p21 Signal Axis / 中国实验方剂学杂志

ObjectiveTo study the effect of Qizhu Kang'ai prescription （QZAP） on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 （PCK1） in the liver of mouse model of liver cancer induced by diethylnitrosamine （DEN） combined with carbon tetrachloride （CCl4） and Huh7 cells of human liver cancer， so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl4 was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group， a model group， and a QZAP group were set up， in which QZAP （3.51 g·kg-1） or an equal volume of normal saline was administered daily by gavage， respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， and alpha-fetoprotein （AFP） in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin （HE） staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment， Huh7 cells were divided into blank group， QZAP low， medium， and high dose groups and/or PCK1 inhibitor （SKF-34288 hydrochloride） group， and Sorafenib group. The corresponding drug-containing serum and drug treatment were given， respectively. Cell counting kit-8 （CCK-8） method， colony formation experiment， Edu fluorescent labeling detection， intracellular adenosine triphosphate （ATP） content detection， and cell cycle flow cytometry detection were used to evaluate the proliferation ability， energy metabolism changes， and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1， serine/threonine kinase （Akt）， phosphorylated Akt （p-Akt）， and cell cycle-dependent protein kinase inhibitor 1A （p21）. ResultCompared with the model group， the pathological changes such as cell atypia， necrosis， and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated， and the number of liver tumors was reduced （P<0.01）. The serum ALT， AST， γ-GT， and AFP levels were reduced （P<0.01）. At the cell level， compared with the blank group， low， medium， and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells （P<0.01） and the number of positive cells with Edu labeling （P<0.01） and inhibit clonal proliferation ability （P<0.01）. The QZAP groups could also reduce the intracellular ATP content （P<0.05） and increase the distribution ratio of the G0/G1 phase of the cell cycle （P<0.05） in a dose-dependent manner. Compared with the model group and blank group， PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced （P<0.05，P<0.01）， and the p-Akt protein level was significantly increased （P<0.01）. Compared with the blank group， the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased （P<0.05）， but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G0/G1 phase distribution. Compared with the SKF-34288 hydrochloride group， the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content， cell survival rate， and Edu-positive cell ratio of Huh7 cells （P<0.05） and significantly increased the G0/G1 phase distribution proportion （P<0.05）. ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression， thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.

Qizhu Anti-Cancer Recipe promotes anoikis of hepatocellular carcinoma cells by activating the c-Jun N-terminal kinase pathway.

Background: Qizhu Anti-Cancer Recipe (QACR) is a traditional Chinese medicine widely used in treating several liver diseases. However, its function and the relevant mechanism underlying its effect in treating hepatocellular carcinoma (HCC) remain unknown. The aim of this study was to explore the effect of QACR in HCC, which are expected to be a potential therapeutic scheme for HCC. Materials and methods: The chemical compositions of QACR were determined by liquid chromatography/quadrupole time-of-fight mass spectrometry (LC-QTOF-MS). The anoikis-resistant HCC cell proliferation and angiopoiesis were detected using the cell counting kit 8 (CCK8) assay, trypan blue, calcein AM/EthD-1, flow cytometer, Western blot, and tube formation assays. An orthotopic xenograft mouse model was established to evaluate the in vivo effects of the QACR. The expression of proliferating cell nuclear antigen (PCNA), Bcl-2, CD31, caspase-3, caspase-8, caspase-9, PARP-1, DFF40, phospho-c-Jun NH2-terminal kinase (p-JNK), and JNK was assessed using Western blot and immunohistochemical analysis. Results: QACR reduced the growth and tube formation of anoikis-resistant HCC cells and enhanced cell apoptosis in vitro. In the orthotopic xenograft mouse models, QACR suppressed the tumorigenesis of HCC in vivo. Mechanistically, QACR modulated the JNK pathway. The JNK inhibitor (SP600125) reverses the inhibitory effects of QACR on anoikis-resistant HCC cell proliferation and angiopoiesis. Conclusion: Our study suggests that QACR suppresses the proliferation and angiopoiesis of anoikis-resistant HCC cells by activating the JNK pathway. Therefore, QACR is a promising new therapeutic strategy for treating hepatocellular carcinoma.

The molecular mechanism of Qizhu anti-cancer prescription in treatment of primary liver cancer based on network pharmacology and experimental verification / 中国药理学通报

Aim To investigate the mechanism of Qizhu anti-cancer prescription ( QZACP) inthe treatment of primary liver cancer using network pharmacology and molecular docking. Methods Drugs and primary liver cancer ( PLC) -related targets were found according to TCMSP database and disease databases such as GeneCard, the key chemical components and core targets were screened by Cytoscape 3. 9. 1 and String platform respectively, and a network relationship diagram of traditional Chinese medicine-active component-target was constructed by using Cytoscape 3.9. 1. GO functional analysis and KEGG pathway analysis were performed using DAVID platform, visualized by R 4. 1. 1 software, and finally the core clustered proteins were analyzed by CytoNCA plug-in to obtain the core action targets, and the core components and key targets were verified by using molecular docking technology and the pharmacodynamic mechanism of QZACP was further verified by animal experiments. Results The active ingredients of QZACP in the treatment of primary liver cancer may be quercetin, glycyrrhizin, Denudatin B, isoflavanone, sanguinarol, etc. ; the potential targets were STAT3, EGFR, AKT1 etc. ; the related pathways were mainly PI3K-Akt signaling pathway,MAPK signaling pathway,etc. ; molecular docking showed that the core compounds had better integrating conformation with the key targets. In addition, QZACP could inhibit the growth of tumor in nude mice and decrease the expression of STAT3, EGFR and AKT1. Conclusions Qizhu anti-cancer prescription may have some positive significance in the treatment of primary liver cancer, which may be related to the regulation of PI3K/Akt signaling pathway.

Herbal Textual Research on Qizhu / 中国实验方剂学杂志

Qizhu, the dried rhizome of Atractylodes macrocephala in Compositae family, is the representative wild variety of Atractylodis Macrocephalae Rhizoma (Baizhu) with modern excellent quality. Through textual research of materia medica works and modern studies, the medication methods between Qizhu and ancient Baizhu were systematically compared. Focusing on seven key issues, this paper systematically summarized the medicinal history, characters, cultivation and other related contents of Qizhu, in order to provide a basis of Qizhu in the recovery and development of its own Daodi-status, and further serve the industrial development of this herb. The name, harvesting time, processing method and other issues had undergone a relatively complicated evolution process. At present, acknowledged points are as following：①The distribution areas of Qizhu include southern areas of the Yangtze River in Anhui province and its surrounding regions. ②Harvesting time is late October. ③Qizhu can be dried in the shade or micro-hot dried after being wrapped with absorbent paper, later it can be divided into two commercial specifications. ④In addition to cutting, there is still a lack of other processing methods. ⑤The superior characters of Qizhu contain white, less oil, fragrant smell and sweet taste and so on. ⑥The history of Qizhu as a genuine medicinal material can be traced back to the Ming dynasty.

Network Pharmacology Approach to Explore Mechanisms of Qizhu Granules for Treatment of Diabetic Nephropathy / 中国实验方剂学杂志

Objective:To explore the mechanism of Qizhu granules in the treatment of diabetic nephropathy by using network pharmacology. Method:The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database （TCMSP） and The Encyclopedia of Traditional Chinese Medicine（ETCM） database were used to screen out the chemical constituents and protein targets of each drug in the Qizhu granules based on oral bioavailability and drug-like properties. The protein target was standardized into the corresponding gene name through the UniProt database. Online Mendelian Inheritance in Man（OMIM）， DisGeNET， Therapeutic Target Database （TTD）， ETCM database were used to search for related targets of diabetic nephropathy， after the intersection of the two， construct a protein interaction network through protein interaction database （STRING）， use Cytoscape to analyze the core target of the network， and the relevant targets were analyzed by KOBAS 3.0 database for Gene Ontology （GO） pathway enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. Result:A total of 93 chemical components were obtained from Qizhu granules， involving 254 targets， and 607 targets related to diabetic nephropathy. After the intersection， 76 sputum granules were determined to treat diabetic nephropathy， including protein kinase B1 （Akt1）， vascular endothelial growth factor （VEGFA）， interleukin （IL）-6， tumor necrosis factor （TNF）， mitogen-activated protein kinase 1 （MAPK1）， matrix metalloproteinase （MMP）-9 and other core targets， after GO analysis and KEGG analysis， Qizhu granules can affect cellular response to nitrogen compound， regulation of reactive oxygen species metabolic process and other biological processes， regulate advanced glycation end product （AGE）/advanced glycation end product receptor （RAGE） signaling pathway in diabetic complications， fluid shear stress and atherosclerosis， IL-17 signaling pathways， HIF-1 signaling pathways TNF signaling pathways and other pathways. Conclusion:The therapeutic effect of Qizhu granules on diabetic nephropathy may affect Akt1，VEGFA， IL-6， TNF， MAPK1， MMP-9 and other targets， and regulate AGE/RAGE signaling pathway in diabetic complications， fluid shear stress and atherosclerosis， IL-17 signaling pathways， hypoxia-inducing factor-1（HIF-1）signaling pathways TNF signaling pathways and other pathways， which can provide a theoretical reference for further basic experimental research.

A preliminary study on quality standard and stability forQizhu-Fuzheng Yin / 国际中医中药杂志

Objective To establish the quality standard ofQizhu-Fuzheng Yin, and to conduct a preliminary study on its stability.Methods Corydalis tuber and licorice were identified by TLC. The content of astragaloside was determined by UPLC-ELSD. The initial stability was studied by accelerated test method. Results The spots on TLC plates were clear without interference in the blank reference. The response of astragaloside was linear in the ranges of 36.5-365.0 μg/ml (r2=0.999 2), and the average recovery was 102.8 %, and theRSD was 2.4%. After 1, 2, 3, 6 months tests, the average contents of 3 batches astragaloside were 61.6, 60.4, 60.6μg/ml.ConclusionQizhu-Fuzheng Yin was simple preparation, quality control and stability.

Study on Therapeutic Effect and Mechanism ofQi-Zhu Er-Zhu Er-CaoTang in Treatment of Chronic Atrophic Gastritis Precancerous Lesion Based on Pathway of NF-κB (p65) and CyclinE / 世界科学技术-中医药现代化

This study was aimed to observe the therapeutic effect and mechanism ofQi-Zhu Er-Zhu Er-Cao Tang(QZEZECT) in the treatment of chronic atrophic gastritis (CAG) precancerous lesion. A total of 56 clean grade healthy Wistar rats were randomly divided into 6 groups. In the negative control group (NC), model group,Wei-Fu-Chun(WFC) group,Ren-Zhu Jian-Wei Ke-Li(RZJWKL, RZ) group, there were 10 rats in each group. In the high-dose QZEZECT (QZ-H) group and low-dose QZEZECT (QZ-L) group, there were 8 rats in each group. CAG/PLGC model was established by MNNG in the model group, WFC group, RZ group, QZ-H group and QZ-L group. Intragastric administration of corresponding decoctions at the dose of 0.39 g·kg-1, 3.2 g·kg-1, 54.4 g·kg-1, 13.6 g·kg-1 were given to rats in the WFC, RZ, QZ-H and QZ-L groups once a day for 28 consecutive days, respectively. The same volume of normal saline was given to the NC group and the model group. Histomorphological changes of gastric mucous membrane in rats of each group were observed. Expressions of NF-κB/p65 and CyclinE protein were detected. The results showed that compared with the NC group, the degrees of infiltration and dysplasia and expressions of NF-κB/p65 and CyclinE significantly increased in the model group with statistical significance (P < 0.01). Compared with the model group, the expression of NF-κB/p65 and CyclinE in the WFC group reduced with statistical significance (P < 0.05). The expression of NF-κB/p65 and CyclinE in the RZ and QZ-L group obviously reduced with statistical significance (P < 0.01). The overall effective rates of WFC group, RZ group, QZ-H group and QZ-L group were 50%, 63.3%, 43.3% and 73.3%, respectively. It was concluded that QZEZECT can treat CAG precancerous lesion, which may take effect by improving and inhibiting pathologic changes of the transcription factor of inflammation.

Inhibitory Effects ofQi-Zhu Granules on Early Proteinuria in Diabetic Nephropathy Rats with Syndrome ofQi-yin Deficiency and Phlegm Blocking Collaterals / 世界科学技术-中医药现代化

ThisstudywasaimedtoobservetheinhibitoryeffectsofQi-Zhu (QZ)granulesonearlyproteinuria in diabetic nephropathy rats with syndrome of qi-yin deficiency and phlegm blocking collaterals. A total of 44 rats were randomly divided into the blank group, model group, Huang-Kui capsule group, QZ granules group. The rat model of diabetic nephropathy with syndrome of qi-yin deficiency and phlegm blocking collaterals was induced by the combination of unilateral renal artery ligation, diet of high-calorie and high cholesterol, and intraperitoneal injection of low-dose streptozotocin. The medication was given for 8 weeks. The concentrations of protein and creatinine in urine were observed on the 4th week. The blood glucose, blood lipids, liver function and renal pathological changes were observed at the end of the experiment. The results showed that compared with the model group, QZ granules can obvious suppress early proteinuria in diabetic nephropathy, promote creatinine excretion, regulate blood lipid metabolism, protect liver function and improve renal pathological changes. It was concluded that QZ granules had independent inhibition effect on early proteinuria in diabetic nephropathy rats with syndrome of qi-yin deficiency and phlegm blocking collaterals. The effect was independent of lowing blood glucose. It represented the corresponding relation between the syndrome and efficacy in Chinese herb compounds.

Establishment of the Method for Testing Leguminosae in Qizhu Bishi Granules / 中国药房

OBJECTIVE:To establish a method for the identification and content determination of leguminosae in Qizhu bishi granules. METHODS:Thin layer chromatography (TLC) was used to identify the leguminosae. HPLC-ELSD was used to deter-mine the content of astragaloside A in leguminosae. The column was Diamonsil C18 with the mobile phase of acetonitrile-water(32∶68,V/V)at the flow rate of 1.0 ml/min,the column temperature was 40 ℃. The sample size was 20 μl. The drift tube temperature was 108 ℃ with the nitrogen gas flow rate of 2.8 ml/min. RESULTS:The TLC features of leguminosae were obvious with clear spots and good specificity. Astragaloside A had a good linear relationship in the range of 0.402 4-4.02 μg(r=0.997 0). RSDs of pre-cision,stability and reproducibility tests were≤0.8%. The average recovery was 99.00%(RSD=1.29%,n=9). CONCLUSIONS:The method is simple,accurate and reliable and can be used for the quality control of Qizhu bishi granules.

Experimental study on Qizhu formula for modulating survivin, an inhibitor of apoptosis, in MGC-803 gastric cancer cells.

The aim of this study was to investigate the mechanism of action of Qizhu formula, a Chinese medicinal empirical formula, in modulating the action of survivin, an inhibitor of apoptosis, in MGC-803 gastric cancer cells. Western blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect the effect of varying concentrations of Qizhu formula in the modulation of the expression of survivin in MGC-803 human gastric adenocarcinoma cells. The western blot analysis results demonstrated that Qizhu formula exerted no significant effects on the protein expression of the ß-actin housekeeping gene, whereas it exerted a significant inhibitory effect on the protein expression of the apoptosis-related survivin gene at concentrations of 250 µg/ml and, particularly, 500 µg/ml. RT-PCR was used to detect the effect of Qizhu formula on survivin mRNA in MGC-803 human gastric adenocarcinoma cells. The ratio of survivin/ß-actin in the 0.1% dimethylsulfoxide and the 125, 250 and 500 µg/ml groups of Qizhu formula was 0.4543, 0.4025, 0.2415 and 0.2235, respectively. Therefore, Qizhu formula exerted a distinct inhibitory effect on the mRNA expression of survivin in MGC-803 cells in a dose-dependent manner. In conclusion, Qizhu formula may modulate the apoptosis of MGC-803 human gastric adenocarcinoma cells, which is associated with the downregulation of survivin mRNA and protein expression.

Influence of rough extraction of QiZhu decoction on VEGF& KDR/flk-1 protein expression in MGC-803 cell / 国际中医中药杂志

Objective To evaluate the influence Qizhu decoction on VEGF and its receptor KDR/flk-1 protein expression in MGC803 cell. Methods Four groups of MGC-803 Cells were established, and intervened with blank control, DMSO control, high dose Qizhu decoction, and low dose Qizhu decoction respectively. VEGF and KDR/flk-1 protein were detected by flow cytometry and western bloting. Results The value of VEGF expression was 120.0±10.8, 116.8±14.7, 95.0±12.5, and 108.4±13.5 respectively in each group. The difference between high dosage Qizhu decoction group and the bland control group was significant. Value of KDR/flk-1 were 10.4±3.5, 9.0±3.4, 6.8±2.3, and 6.8±3.5 in each group respectively. There was no significant difference among these 4 groups. The grayseale values of VEGF expression was 14.45±4.61, 12.32±3.27, 2.58±0.84, and 3.45±1.12 in each group respectively. There was significant difference between the QiZhu groups and the blank control group. Meanwhile grayseale values of KDR/flk-1 expression were 3.87±1.05, 3.55±1.32, 3.62±1.01, and 3.73±0.88 in each group respectively, showing no significant difference among 4 groups. Conclusion Rough extraction of QiZhu decoction down-regulated the protein expression of VEGF, but had no effects on the expression of KDR/flk-1.

Experimental Study on Qizhu Jianzhong Decoction Influencing Ulcerative Rats’ IL-6 and GAS in Spleen Yang Deficiency / 浙江中医药大学学报

[Objective] To explore part of mechanism of Qizhu Jianzhong Decoction treating gastric ulcer and spleen Yang deficiency, impersonally state the scientific connotation of the decoction. [Method] Randomly divide 60 SD rats into normal/ blank control group(Ⅰ), model group(Ⅱ), sucralfate group(Ⅲ), large-dosage group(Ⅳ), middle-dosage group(Ⅴ) and small-dosage group(Ⅵ) of Qizhu Jianzhong Decoction. Except for the group Ⅰ, others are made into rats model of spleen yang deficiency and gastric ulcer with abnormal diet, fatigue, cold and bitter purgation, and cold water stimulant. After being administered medicines, measure their IL-6 and GAS level, and observe the pathological changes of gastric mucosa under microscope. [Result] Compared with group Ⅰ, in the model group, the IL-6 rose, GAS decreased; compared with the model group, other groups of Ⅲ, Ⅳ and Ⅴ all markedly decreased rats serum Ⅱ-6 level and increased GAS level, with obvious difference(P0.05). Under microscope, the gastric mucosa injury degree was less in groups Ⅲ, Ⅳ and Ⅴ than group Ⅱ; and it was similar between group Ⅵ and group Ⅱ. [Conclusion] Groups Ⅳ and Ⅴ have marked cure effect on spleen yang deficiency and gastric ulcer, and group Ⅵ has no obvious cure effect.

Effect of Qizhu Oral Liquid on radiation injury in mice / 中成药

AIM: To observe the protective effect of Qizhu Oral Liquid on radiation injury in mice. METHODS: The model was made in mice by X-ray radiation. The white blood cells(WBC), bone marrow nucleated cells (BMNC) were measured. The weight of thymus and spleen were measured. RESULTS: Qizhu Oral Liquid could increase WBC and BMNC remarkably. It could increase the weight of thymus and spleen of mice signifcantly. CONCLUSION: Qizhu Oral Liquid is effective in protecting the X-radiation injury mice.