Multi-center Randomized Controlled Clinical Trial of Huangqi Injection Combined with Buzhong Yiqi Acupuncture in Treatment of Chronic Fatigue Syndrome with Qi Deficiency / 中国实验方剂学杂志

ObjectiveTo investigate the clinical efficacy of Huangqi injection combined with Buzhong Yiqi acupuncture in the treatment of chronic fatigue syndrome （CFS） with Qi deficiency and its effects on TCM syndromes， fatigue symptoms， serum superoxide dismutase （SOD）， malondialdehyde （MDA）， and oxidized low-density lipoprotein （ox-LDL） levels. MethodA total of 200 patients with CFS of Qi deficiency were randomly divided into a control group （100 cases） and an observation group （100 cases）. The control group was treated with vitamin B compounds， and the observation group was treated with Huangqi injection combined with Buzhong Yiqi acupuncture for two weeks. The scores of TCM syndromes， fatigue symptoms， levels of serum SOD， MDA， and ox-LDL and the incidence of adverse reactions were observed and compared before and after treatment in two groups. ResultAfter treatment， the total effective rate of the control group was 54.34% （50/92）， while that of the observation group was 88.54% （85/96）. The total effective rate of the observation group was higher than that of the control group （χ2=27.13，P<0.05）. Compared with those in the two groups before treatment， scores of fatigue self-assessment scale （FSAS）， physical fatigue and mental fatigue， and sleep/rest response scores of fatigue in the two groups after treatment were significantly decreased （P<0.05）. After treatment， scores of FSAS， physical fatigue and mental fatigue， and sleep/rest response scores of fatigue in the observation group were significantly decreased compared with those in the control group （P<0.05）. Compared with those in the two groups before treatment， TCM syndrome scores in the two groups after treatment were significantly decreased （P<0.05）. After treatment， TCM syndrome scores in the observation group were significantly decreased compared with those in the control group （P<0.05）. Compared with those in the two groups before treatment， MDA levels in the two groups were significantly decreased （P<0.05）， ox-LDL levels in the observation group were significantly decreased （P<0.05）， and SOD levels were significantly increased （P<0.05）. After treatment， compared with those in the control group， the serum MDA and ox-LDL levels in the observation group were significantly decreased （P<0.05）， and the serum SOD was significantly increased （P<0.05）. No serious adverse events or adverse reactions occurred during this clinical trial. ConclusionHuangqi injection combined with Buzhong Yiqi acupuncture has a good clinical curative effect in the treatment of CFS with Qi deficiency， which can effectively improve the fatigue symptoms of patients， increase the level of SOD， and reduce the level of serum MDA and ox-LDL. It is related to the production of antioxidants， inhibiting the production of lipid peroxides， and improving the body's ability to resist oxidative stress.

Protective Effects and Mechanisms of Huangqi Decoction Against Radiation-Induced Bystander Effects / Efectos Protectores y Mecanismos de la Decocción de Huangqi Contra los Efectos Secundarios Inducidos por la Radiación

SUMMARY: The 12C6+ heavy ion beam irradiation can cause bystander effects. The inflammatory cytokines, endocrine hormones and apoptotic proteins may be involved in 12C6+ irradiation-induced bystander effects. This study characterized the protective effects and mechanisms of Huangqi decoction (HQD) against 12C6+ radiation induced bystander effects. Wistar rats were randomly divided into control, 12C6+ heavy ion irradiation model, and high-dose/medium-dose/low-dose HQD groups. HE staining assessed the pathological changes of brain and kidney. Peripheral blood chemical indicators as well as inflammatory factors and endocrine hormones were detected. Apoptosis was measured with TUNEL. Proliferating cell nuclear antigen (PCNA) expression was determined with real-time PCR and Western blot.Irradiation induced pathological damage to the brain and kidney tissues. After irradiation, the numbers of white blood cells (WBC) and monocyte, and the expression of interleukin (IL)-2, corticotropin-releasing hormone (CRH) and PCNA decreased. The damage was accompanied by increased expression of IL-1β, IL-6, corticosterone (CORT) and adrenocorticotropic hormone (ACTH) as well as increased neuronal apoptosis. These effects were indicative of radiation-induced bystander effects. Administration of HQD attenuated the pathological damage to brain and kidney tissues, and increased the numbers of WBC, neutrophils, lymphocyte and monocytes, as well as the expression of IL-2, CRH and PCNA. It also decreased the expression of IL-1β, IL-6, CORT and ACTH as well as neuronal apoptosis. HQD exhibits protective effects against 12C6+ radiation-induced bystander effects. The underlying mechanism may involve the promotion of the production of peripheral blood cells, inhibition of inflammatory factors and apoptosis, and regulation of endocrine hormones.

La irradiación con haz de iones pesados 12C6+ puede provocar efectos secundarios. Las citoquinas inflamatorias, las hormonas endocrinas y las proteínas apoptóticas pueden estar involucradas en los efectos secundarios inducidos por la irradiación 12C6+. Este estudio caracterizó los efectos y mecanismos protectores de la decocción de Huangqi (HQD) contra los efectos externos inducidos por la radiación 12C6+. Las ratas Wistar se dividieron aleatoriamente en grupos control, modelo de irradiación de iones pesados 12C6+ y grupos de dosis alta/media/baja de HQD. La tinción con HE evaluó los cambios patológicos del cerebro y el riñón. Se detectaron indicadores químicos de sangre periférica, así como factores inflamatorios y hormonas endocrinas. La apoptosis se midió con TUNEL. La expresión del antígeno nuclear de células en proliferación (PCNA) se determinó mediante PCR en tiempo real y transferencia Western blot. La irradiación indujo daños patológicos en los tejidos cerebrales y renales. Después de la irradiación, disminuyó el número de glóbulos blancos (WBC) y monocitos, y la expresión de interleucina (IL)-2, hormona liberadora de corticotropina (CRH) y PCNA. El daño estuvo acompañado por una mayor expresión de IL-1β, IL-6, corticosterona (CORT) y hormona adrenocorticotrópica (ACTH), así como un aumento de la apoptosis neuronal. Estas alteraciones fueron indicativas de efectos inducidos por la radiación. La administración de HQD atenuó el daño patológico a los tejidos cerebrales y renales, y aumentó el número de leucocitos y monocitos, así como la expresión de IL-2, CRH y PCNA. También disminuyó la expresión de IL-1β, IL-6, CORT y ACTH, así como la apoptosis neuronal. HQD exhibe mecanismos protectores contra los efectos externos inducidos por la radiación 12C6+. El mecanismo subyacente puede implicar la promoción de la producción de células sanguíneas periféricas, la inhibición de factores inflamatorios y la apoptosis y la regulación de hormonas endocrinas.

Research progress on Huangqi Guizhi Wuwu Decoction and predictive analysis of quality markers / 中国中药杂志

Huangqi Guizhi Wuwu Decoction is a classic prescription in traditional Chinese medicine(TCM) and is known for its effects of tonifying Qi, warming the meridians, and promoting blood circulation to alleviate obstruction. It is primarily used to treat conditions characterized by Qi stagnation, Yang deficiency, and obstruction, and it exhibits pharmacological effects such as immune regulation, anti-inflammation, analgesia, protection of the cardiovascular and cerebrovascular systems, itch relief, reduction of frostbite symptoms, antioxidative stress, promotion of cell apoptosis, and kidney protection. In modern clinical practice, it is commonly used to treat acute myocardial infarction, sequelae of cerebral infarction, cervical spondylosis, frozen shoulder, lower limb arteriosclerosis, lower limb vascular disorders, peripheral neuropathy in diabetes, and lupus nephritis. Recent research has focused on the chemical components, pharmacological effects, and clinical applications of Huangqi Guizhi Wuwu Decoction. Based on the "five principles" of quality markers(Q-markers) in TCM, this study predicted and analyzed the Q-markers of Huangqi Guizhi Wuwu Decoction. It suggested that astragaloside Ⅳ, formononetin, kaempferol, quercetin, cinnamic acid, cinnamaldehyde, 6-gingerol, paeoniflorin, albiflorin, and gallic acid could serve as Q-markers for Huangqi Guizhi Wuwu Decoction. The findings of this study can provide references for quality control of Huangqi Guizhi Wuwu Decoction and the development of new Chinese medicinal formulations.

Protective Effect of Huangqi Baihe Granules on Acute Brain Injury in Rats with High Altitude Hypoxia Based on HIF-1α/NF-κB/NLRP3 Signal Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α （HIF-1α）/nuclear factor-κB （NF-κB）/NOD-like receptor hot protein domain related protein 3 （NLRP3） signaling pathway in a rat model of high altitude hypoxia. MethodSixty male SPF SD rats were randomly divided into blank group， model group， dexamethasone group （5 mg·kg-1）， and high， middle， and low-dose groups of Huangqi Baihe granules （4.1， 2.05， 1.025 g·kg-1）. Among them， each Chinese medicine group was administrated orally for continuously 14 d， once a day， and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group. On the 15th d， the model group， dexamethasone group， and high， middle， and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude， low pressure， and low oxygen environment in the animal low-pressure simulation cabin， and the exposure lasted for 3 d. Blood was collected from the abdominal aorta and serum was separated， and the brain tissue was taken after being killed. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in brain tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in rat serum. Western blot was used to detect HIF-1α， NLRP3， phosphorylated nuclear factor-κB （p-NF-κB）， NF-κB， desquamation D （GSDMD）， and cysteine aspartate-specitis protein-1（Caspase-1） in rats of each group. The mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultThe results of HE staining showed that as compared with the normal group， the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder， with different sizes. Compared with the model group， the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules were reduced. The results of ELISA showed that as compared with the normal group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the model group was significantly higher （P<0.01）. Compared with the model group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly （P<0.05， P<0.01）. The results of Western blot showed that as compared with the normal group， the relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly higher （P<0.01）. As compared with the model group， the relative expressions of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased （P<0.05， P<0.01）. The relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly （P<0.01）， and the relative protein expression of HIF-1α in the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced （P<0.05）. The Real-time PCR analysis showed that as compared with the normal group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly increased （P<0.01）. As compared with the model group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased （P<0.01）. The mRNA expression levels of HIF-1α， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly （P<0.01）. The mRNA expression levels of HIF-1α， NLRP3， and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased （P<0.05， P<0.01）. ConclusionThe protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway.

Study on Huangqi Jiedu Decoction for Wnt/β-catanin signaling pathway inhibiting EMT against lung metastasis of breast cancer / 国际中医中药杂志

Objective:To observe the inhibitory effects of Huangqi Jiedu Decoction on lung metastasis of breast cancer in nude mice; To explore the mechanism of intervening epithelial mesenchymal transformation (EMT) induced by Wnt/β-catenin signaling pathway.Methods:Totally 30 nude mice were divided into model group, adriamycin group and Huangqi Jiedu Decoction low-, medium-, and high-dosage groups according to random number table method. Each group was injected subcutaneously with mouse breast cancer 4T1 cells to construct tumor - bearing nude mice model. Huangqi Jiedu Decoction low-, medium- and high-dosage groups were intragastrically administrated with Huangqi Jiedu Decoction 17.82, 35.64 and 71.28 g/kg; adriamycin group was injected intraperitoneally adriamycin 0.05 g/kg; model group was intragastrically administrated with normal saline of the same volume for 21 d. Tumor volume was measured at 9, 15, and 21 days after modeling. After the end of administration, the tumor tissue was separated, the tumor weight was measured, and the tumor inhibition rate was calculated. The lung tissue was Isolated,, the number of lung metastatic nodules and the inhibition rate of lung metastasis was counted. HE staining was used to observe the tissue morphology and evaluate the effectiveness of the model. The protein expressions of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by Western Blot. The mRNA levels of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by real-time fluorescent quantitative PCR.Results:Compared with the model group, the tumor volume and mass of Huangqi Jiedu Decoction low-, medium- and high-dosage groups decreased ( P<0.01); the number of pulmonary metastasis nodules in Huangqi Jiedu Decoction high-dosage group significantly decreased ( P<0.01); the mRNA and protein expressions of β-catenin and Vimentinm decreased in the Huangqi Jiedu Decoction low-, medium- and high-dosage groups ( P<0.01), and the protein and mRNA expressions of E-Cadherin increased in the Huangqi Jiedu Decoction high-dosage group ( P<0.01). Conclusion:Huangqi Jiedu Decoction can effectively inhibit the growth and lung metastasis of breast cancer transplanted tumor, and the mechanism may be to down-regulate the expression of key molecules in the Wnt/β-catanin signaling pathway, thereby inhibiting the EMT process, so as to inhibit the lung metastasis of breast cancer.

Effect of Neibu Huangqi Decoction combined with Kangfuxin Liquid on wound healing after hemorrhoid fistula / 国际中医中药杂志

Objective:To investigate the effects of Neibu Huangqi Decoction combined with Kangfuxin Liquid on wound healing after hemorrhoid fistula.Methods:Randomized controlled trial. A total of 90 patients with hemorrhoid fistula surgery in Tangshan Hospital of Traditional Chinese Medicine from January 2020 to June 2021 were selected as the observation objects and divided into 2 groups by random number table method, with 45 cases in each group. The control group was treated with Kangfuxin Liquid after surgery, and the observation group was treated with Neibu Huangqi Decoction. Both groups were treated continuously for 14 days. Wound symptom score was performed before and after treatment. The levels of TNF-α, IL-6 and IL-8 were determined by ELISA. The wound healing time was observed and the wound healing rate was calculated. Adverse reactions were recorded and clinical efficacy was evaluated.Results:The total effective rate was 93.33% (42/45) in the observation group and 66.67% (30/45) in the control group, with statistical significance ( χ2=9.89, P=0.002). After treatment, the scores of pain [(0.63±0.14) vs. (0.97±0.27), t=7.50], exudation [(0.67±0.12) vs. (1.09±0.31), t=8.48], edema [(0.78±0.17) vs.(1.25±0.36), t=7.92], pruritus [(0.78±0.20) vs. (1.32±0.33), t=9.39] were lower than those in the control group ( P<0.01); serum TNF-α [(33.46±2.86) μg/L vs. (45.78±3.92) μg/L, t=25.39], IL-6 [(41.86±5.84) μg/L vs. (56.12±6.75) μg/L, t=15.98], IL-8 [(27.40±3.58) ng/L vs. (36.16±3.84) ng/L, t=16.69] were lower than those in the control group ( P<0.01). The wound healing time of the observation group was shorter than that of the control group ( t=8.60, P<0.01), and the wound healing rate was higher than that of the control group ( t=24.65, P<0.01). During treatment, the incidence of adverse reactions was 11.11% (5/45) in the observation group and 6.67% (3/45) in the control group, without statistical significance ( χ2=0.14, P=0.711). Conclusion:Neibu Huangqi Decoction combined with Kangfuxin Liquid can promote wound healing, reduce inflammatory cytokines, relieve pain and exudation, improve clinical efficacy, and have few adverse reactions.

Clinical effect of Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction in the treatment of ischemic stroke during recovery / 国际中医中药杂志

Objective:To evaluate the clinical curative effect of Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction on patients with ischemic stroke (IS) during recovery.Methods:Prospective cohort study. A total of 220 patients with IS during recovery who met inclusion criteria in the First People's Hospital of Dongcheng District in Beijing, were enrolled and divided into control group ( n=110) and observation group ( n=110) by random number table method between January 2015 and July 2020. The control group was given basic treatment, while observation group was given Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction. All were treated for 1 month. Before and after treatment, Traditional Chinese Medicine (TCM) syndromes were scored. The activities of daily living were evaluated by Barthel index. The quality of life was evaluated by stroke specific quality of life scale (SS-QOL). The severity of neurological impairment was evaluated by National Institute of Health Stroke Scale (NIHSS). The functional recovery of stroke was evaluated by modified Rankin scale (mRS). The adverse reactions during treatment were observed and recorded. Results:After treatment, scores of TCM syndromes, mRS and NIHSS in observation group were significantly lower than those in the control group ( t=21.87, 4.66, 12.06, P<0.01), while scores of Barthel index and SS-QOL were significantly higher than those in the control group ( t=14.13, 5.80, P<0.01). During treatment, there were no obvious adverse reactions in either group. Conclusion:Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction can improve clinical symptoms and nerve function in patients with IS during recovery, which is beneficial to improve outcomes and quality of life.

Effect of Modified Huangqi Guizhi Wuwutang on Apoptosis-related Bax and Caspase-12 of Sciatic Nerve Cells in Diabetes Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effect of modified Huangqi Guizhi Wuwutang （MHGW） on the protein and mRNA expression of B-cell lymphoma-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase-12 （Caspase-12） related to the apoptosis of sciatic nerve cells in diabetes rats to explore the mechanism of MHGW in the treatment of peripheral neuropathy in diabetes. MethodAnimal experiments were conducted. A diabetes model was induced in sixty male sprague-dawley （SD） rats by feeding on a high-sugar and high-fat diet combined with streptozotocin （STZ） intraperitoneal injection. Rats with random blood glucose levels ≥ 16.7 mmol·L-1 for three consecutive days were considered to have successfully developed diabetes. Forty-eight rats that successfully developed diabetes were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg-1·d-1）， a high-dose MHGW group （2.5 g·kg-1·d-1）， and a low-dose MHGW group （1.25 g·kg-1·d-1）， with 12 rats in each group. Another 10 rats were assigned to the normal group. Body weight and random blood glucose levels of the rats were monitored. At the end of a 16-week intervention period， the sciatic nerve conduction velocity of the rats was measured using the Key point electromyography collection system. The protein and mRNA expression of Bax and Caspase-12 in the sciatic nerve cells was detected by Western blot analysis and real-time quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultCompared with the normal group， the model group showed a significant decrease in body weight （P<0.01） and a significant increase in random blood glucose levels （P<0.01）. After a 16-week intervention， compared with the model group， the high-dose MHGW group exhibited a significant increase in body weight （P<0.05）， while there were no statistically significant differences in body weight changes among the other treatment groups. Random blood glucose levels significantly decreased in all treatment groups （P<0.01）. After 16 weeks of intervention， compared with the normal group， the model group had significantly reduced motor and sensory nerve conduction velocities （P<0.01）. Compared with the model group， all treatment groups showed significant increases in motor and sensory nerve conduction velocities （P<0.05， P<0.01）. The expression of Bax and Caspase-12 proteins in the sciatic nerve cells was significantly elevated in the model group compared with that in the normal group （P<0.01）. In contrast， all treatment groups showed significant reductions in the expression of Bax and Caspase-12 proteins in the sciatic nerve cells as compared with that in the model group （P<0.01）. The expression of Bax and Caspase-12 mRNA in the sciatic nerve cells significantly increased in the model group compared with that in the normal group （P<0.01）. Compared with the model group， the α-lipoic acid group and the high-dose MHGW group showed significant reductions in the expression of Bax mRNA in the sciatic nerve cells （P<0.05， P<0.01）， while the low-dose MHGW group showed a decreasing trend in the expression of Bax mRNA. The expression of Caspase-12 mRNA in the sciatic nerve cells significantly decreased in all treatment groups （P<0.01）. ConclusionMHGW may improve and repair sciatic nerve damage in diabetes rats by inhibiting sciatic nerve cell apoptosis.

Effect of Modified Huangqi Guizhi Wuwutang on Stress-related IRE1α/CHOP Pathway of Sciatic Nerve Endoplasmic Reticulum in Diabetes Rats / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of modified Huangqi Guizhi Wuwutang （MHGW） on endoplasmic reticulum stress in the sciatic nerve of diabetes rats based on the pathways of inositol-requiring enzyme 1α （IRE1α） and CCAAT/enhancer-binding protein homologous protein （CHOP）. MethodSixty rats were fed on a high-sugar and high-fat diet for six weeks， followed by intraperitoneal injection of streptozotocin at a dose of 35 mg·kg-1. Random blood glucose levels were measured three days later and rats with a sustained blood glucose level ≥ 16.7 mmol·L-1 were included in study （n=48）. The rats were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg-1·d-1）， a high-dose MHGW group （2.5 g·kg-1·d-1）， and a low-dose MHGW group （1.25 g·kg-1·d-1）. Another 10 rats were assigned to the normal group. The intervention lasted for 16 weeks. After 16 weeks， the sciatic nerve structure of the rats in each group was observed under light microscopy using Luxol fast blue （LFB） staining. Transmission electron microscopy was used to observe the ultrastructure of the sciatic nerve. Chemiluminescence method was employed to measure the serum reactive oxygen species （ROS） levels. Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to evaluate the expression of p-IRE1α protein， IRE1α mRNA， CHOP protein， and CHOP mRNA in the sciatic nerve of the rats. ResultCompared with the normal group， the model group showed elevated serum ROS levels （P<0.01）. In contrast， the serum ROS levels were significantly reduced in the treatment groups compared with those in the model group （P<0.01）. The sciatic nerve of the model group showed pathological changes compared with that in the normal group， while the treatment groups exhibited improvement in sciatic nerve pathology compared with the model group. The protein expression of p-IRE1α and CHOP in the sciatic nerve significantly increased in the model group as compared with that in the normal group （P<0.01）. However， the treatment groups showed a significant decrease in the protein expression of p-IRE1α and CHOP in the sciatic nerve compared with the model group （P<0.05， P<0.01）. Furthermore， compared with the normal group， the model group showed upregulated mRNA expression of IRE1α and CHOP in the sciatic nerve （P<0.01）， while the treatment groups exhibited a significant decrease in the mRNA expression of IRE1α and CHOP compared with the model group （P<0.01）. ConclusionMHGW can alleviate endoplasmic reticulum stress-induced cell apoptosis and improve the structure and function of the sciatic nerve in diabetes rats by inhibiting the expression of IRE1α/CHOP pathway-related proteins and mRNA， thereby preventing and treating peripheral neuropathy in diabetes.

Huangqi Decoction, a compound Chinese herbal medicine, inhibits the proliferation and activation of hepatic stellate cells by regulating the long noncoding RNA-C18orf26-1/microRNA-663a/transforming growth factor-β axis / 中西医结合学报

OBJECTIVE@#Huangqi Decoction (HQD), a classical traditional Chinese medicine formula, has been used as a valid treatment for alleviating liver fibrosis; however, the underlying molecular mechanism is still unknown. Although our previous studies showed that microRNA-663a (miR-663a) suppresses the proliferation and activation of hepatic stellate cells (HSCs) and the transforming growth factor-β/small mothers against decapentaplegic (TGF-β/Smad) pathway, whether long noncoding RNAs (lncRNAs) are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported. The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.@*METHODS@#The expression levels of lnc-C18orf26-1, miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction. HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs. The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs. Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs. RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs. Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA. The expression levels of collagen α-2(I) chain (COL1A2), α-smooth muscle actin (α-SMA) and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.@*RESULTS@#Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a. Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation, downregulated TGF-β1-stimulated α-SMA and COL1A2 expression, and inhibited the TGF-β1/Smad signaling pathway. HQD suppressed the proliferation and activation of HSCs. HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs. Further studies showed that HQD inhibited the expression of COL1A2, α-SMA, TGF-β1, TGF-β type I receptor (TGF-βRI) and phosphorylated Smad2 (p-Smad2) in HSCs, and these effects were reversed by miR-663a inhibitor treatment.@*CONCLUSION@#Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis. HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.

The intervention effect of Tibetan medicine Siwei Huangqi powder on hypoxic pulmonary hypertension through autophagy signaling pathway induced by AMPK protein / 中国药理学通报

Aim To explore the mechanism of Tibetan medicine Siwei Huangqi powder(SW)in reducing hypoxic pulmonary hypertension. Methods A total of 110 Wistar rats were randomly divided into normoxia control group,hypoxic control group and hypoxic drug group. The two hypoxic groups were divided into 1,3,7,15 and 30 day group according to the exposure time of hypoxic,10 groups in total. The normoxia control group was placed in the atmospheric environment at an altitude of 2 260 meters without intervention; 10 hypoxic groups were placed in a hypobaric hypoxic chamber with a simulated altitude of 5 000 meters. The hypoxic drug group was given SW suspension(0.42 g/100 g)by gavage,and the hypoxic control group was given normal saline by gavage,once a day. The Ppa and RV/(LV+S)were measured at the corresponding time points in each group; the levels of p-AMPK,ULK-1 and LC3 /LC3 Ⅱ protein in lung tissues were measured by WB method. Results Compared with normoxia control group,the ratio of PA and RV/(LV + s)in hypoxic control group increased gradually with the extension of hypoxic exposure time,which was consistent with the thickness of pulmonary artery smooth muscle layer and the changes of pulmonary tissue subcellular organelles. The expression level of p-AMPK protein in lung tissues was also slightly up-regulated(P<0.05),and ULK-1 and LC3 Ⅱ were significantly up-regulated(P<0.01),especially in acute hypoxic. Compared with the hypoxic control group,the increase of Ppa and the thickening of pulmonary artery smooth muscle layer in the hypoxic drug group were significantly reduced(P<0.050.01),while the expression levels of p-AMPK,ULK-1 and LC3 Ⅱ proteins in lung tissues increased with the extension of hypoxic exposure time(P<0.050.001),especially in chronic hypoxic. Conclusion SW can inhibit hypoxic pulmonary hypertension by up-regulating AMPK autophagy signaling pathway.

Clinical Experience of Treating Metabolic Associated Fatty Liver Disease with Huangqi （Radix Astragali） in Lingnan （岭南） Area / 中医杂志

Based on the unique syndrome characteristics of metabolic associated fatty liver disease （MAFLD） in Lingnan （岭南） area， this paper discussed the clinical experience of Huangqi （Radix Astragali） in the treatment of MAFLD in Lingnan area. It is summarized that spleen deficiency and yang weakness， and internal accumulation of damp-turbidity are the source of the disease， and at the early stage， Huangqi， commonly 15~30 g， could be used to fortify the spleen and warm the earth， thereby making the deficiency fire latent， as well as raise the clear and warm the exterior so as to inhibit damp-turbidity； medication needs to be progressively supplemented， and can be used together with Taizishen （Radix Pseudostellariae）， Baizhu （Rhizoma Atractylodis Macrocephalae）， Fuling （Poria）， and Fangfeng （Radix Saposhnikoviae） to back up the earth and inhibit dampness. Qi stagnation and blood constraint， yin dampness and heat accumulation are the pathogenesis of disease progression. Huangqi should be taken to invigorate the spleen and flourish the liver， unblock yang， and move stagnation. The dosage is often maintained at 30~60 g， to circulate the qi and unblock yang qi in the middle jiao （焦）， usually combined with Chaihu （Radix Bupleuri）， Baishao （Radix Paeoniae Alba） and Yujin （Radix Curcumae） integrating warm and cool medicinals. Stubborn turbid and fat condensing in the liver is a severe stage of the disease and may be concurrent with various pathogens such as dampness， phlegm， and heat， for which 60~90 g Huangqi should be used to invigorate blood and disperse fat， reinforce healthy qi and expel pathogens， often with medicinals that can disperse fat and direct the turbid downward such as Shanzha （Fructus Crataegi）， Juemingzi （Semen Cassiae）， Lulutong （Fructus Liquidambaris） and Zexie （Rhizoma Alismatis）. When prescribing， it is suggested to combine with other medicinals according to the season， thereby adjusting the ascending and descending of the property of Huangqi， thereby conforming to the nature of the four seasons.

Huangqi Sijunzi decoction for treating cancer-related fatigue in breast cancer patients: a randomized trial and network pharmacology study / 南方医科大学学报

OBJECTIVE@#To evaluate the clinical efficacy of Huangqi Sijunzi decoction (HQSJZD) for treating cancer-related fatigue (CRF) of spleen and stomach Qi deficiency type after chemotherapy in patients with breast cancer.@*METHODS@#A total of 94 breast cancer patients who developed CRF of spleen and stomach Qi deficiency type after chemotherapy were randomized into chemotherapy group (n=47) and traditional Chinese medicine (TCM) + chemotherapy group (n=47). The patients in chemotherapy group received the AC or EC regimen and non-drug interventions including psychological counseling, and those in TCM + chemotherapy group received oral administration of HQSJZD in addition to chemotherapy for 21 days as a treatment cycle, after which improvement of fatigue was assessed using Modified Piper Fatigue Scale. The active ingredients and targets of HQSJZD were screened using the TCM System Pharmacology Analysis Platform (TCMSP); the CRF- and breast cancer-related disease targets were retrieved based on data from the GeneCards, NCBI gene and OMIM databases to construct the component-target network and the protein-protein interaction (PPI) network. GO functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes KEGG pathway enrichment analysis of the target genes were performed to construct the component-disease-pathway-target biological network. The binding strength of the major drug ingredients and CRF key targets were predicted using AutoDock software.@*RESULTS@#The scores for somatic fatigue, emotional fatigue and cognitive fatigue, along with the overall fatigue score, showed more significant improvements in TCM+chemotherapy group than in chemotherapy group (P < 0.001), and the response rate reached 89.4% in the combined treatment group. We identified 250 targets for HQSJZD, 2653 CRF-related genes, 15 329 breast cancer-related genes and 161 prescription-disease intersected targets, from which topological analysis identified 66 potential key targets. GO and KEGG enrichment analyses predicted multiple pathways related with the disease. Molecular docking results suggested that the core ingredients of HQSJZD showed high affinities to the key targets AKT1, CASP3, IL6, JUN and VEGFA, among which AKT1 might be the most important target for HQSJZD to treat CRF.@*CONCLUSION@#HQSJZD can obviously improve CRF symptoms in breast cancer patients possibly by regulating multiple signaling pathways including PI3K-Akt through AKT1.

Effect of Huangqi Guizhi Wuwutang on Diabetic Peripheral Neuropathy in MKR Mice via Regulating Endoplasmic Reticulum Stress / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Huangqi Guizhi Wuwutang （HQGZWWT） in the treatment of diabetic peripheral neuropathy （DPN） in MKR mice via regulating endoplasmic reticulum （ER） stress. MethodThirty-two 8-week-old MKR mice （half were male and half were female） were fed with a high-fat diet for four weeks， and then 1% streptozotocin （STZ） was injected intraperitoneally for five days. After the blood glucose was stabilized， the mice were housed in the cage covered with ice bags for another one hour stimulation per day for four weeks. Mice with fasting blood glucose （FBG） value ≥11.1 mmol·L-1 were randomly divided into model group ， Huangqi Guizhi Wuwutang in original dosage group （30 g·kg-1·d-1）， Huangqi Guizhi Wuwutang in formula dosage group （6.25 g·kg-1·d-1）， and positive drug group （mecobalamin tablets， 0.17 mg·kg-1·d-1）. Another eight MKR mice of the same age were set as blank group and eight FVB mice were normal group. After four weeks of intragastric administration in each group， the change in FBG was tested， and hematoxylin and eosin （HE） staining and transmission electron microscope were used for observing the morphology of sciatic nerve tissue. In addition， the expression of c-Jun N-terminal kinase （JNK）， phosphorylated c-Jun N-terminal kinase （p-JNK） and inositol requiring enzyme 1α （IRE1α） proteins was determined by immunohistochemical test and Western blot （WB）. ResultCompared with the conditions in the normal group and blank group， the time of paw withdrawal， paw licking and tail flick in the model group was shortened （P<0.01）， and the conduction velocity of sciatic nerve was decreased （P<0.01）. Compared with the conditions in the model group， the behavioral and functional indicators were improved by HQGZWWT （P<0.05，P<0.01）. The immunohistochemical test revealed the JNK expression was elevated in the model group compared with the conditions in the normal group and blank group （P<0.05）， while that was lowered by HQGZWWT compared with the condition in the model group （P<0.05）. However， there was no difference among the treatment groups. According to the WB， the expression of IRE1α and p-JNK in the model group was enhanced compared with the conditions in the normal group and blank group （P<0.05，P<0.01）， while that was decreased by HQGZWWT compared with the condition in the model group （P<0.05，P<0.01）. No difference was observed between the HQGZWWTO and HQGZWWTF groups. ConclusionHQGZWWT can improve the neurophysiological function and pathological damage of sciatic nerve， which may be related to its delaying the ER stress response of sciatic nerve.

Fangji Huangqi Detumescence Prescription Treats Synovitis in Rats with Knee Osteoarthritis by Modulating Polarization of Synovial Macrophages / 中国实验方剂学杂志

ObjectiveTo assess the curative effects of Fangji Huangqi detumescence prescription （FHDP） on synovitis and polarization of synovial macrophages of knee osteoarthritis （KOA） model in rats induced by Hulth method. MethodThirty-six rats were randomly divided into sham operation group， model group， high-dose， medium-dose， and low-dose （29.16， 14.58， and 7.29 g·kg-1） FHDP groups， and loxoprofen sodium （16.2 mg·kg-1） group. KOA model in rats was induced by modified Hulth method. Six weeks after the operation， rats were given high， medium， and low concentrations of FHDP， normal saline （NS）， and loxoprofen sodium according to the group to intervene， and sacrificed after 2-week administration. Synovium and cartilage histopathological changes were observed after hematoxylin-eosin （HE） staining. Flow cytometry （FCM） and immunofluorescence （IF） test were used to evaluate the polarization of M1/M2 macrophages. Immunohistochemistry （IMC） and enzyme-linked immunosorbent assay （ELISA） were used to detect the related protein expression levels of macrophage polarization， such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， interleukin-10 （IL-10）， and matrix metalloproteinase-13 （MMP-13） in joint tissues and serum. ResultCompared with the sham operation group， Krenn and Mankin scores in the model group were significantly increased （P<0.01）. Compared with the model group， Krenn score was decreased in all administration groups （P<0.05， P<0.01）， but there was no significant difference in Mankin score in any administration groups. Compared with the sham operation group， M1/mø （CD38+） ratio in the model group was significantly increased （P<0.01）， and M2/mø （CD206+） ratio in the model group was decreased （P<0.05）. Compared with the model group， M1/mø ratio in the high， medium， and low-dose FHDP groups was decreased （P<0.05， P<0.01）， but M2/mø ratio was increased in all administration groups （the difference had no statistical significance）. Compared with the sham operation group， M1/M2 ratio in the model group was significantly increased （P<0.01）. Compared with the model group， M1/M2 ratio in all FHDP groups was significantly decreased （P<0.01）， and M1/M2 ratio in the high and medium-dose FHDP groups was lower than that in the loxoprofen sodium group （P<0.05）. Compared with the sham operation group， the levels of TNF-α， IL-1β， and MMP-13 in synovium and cartilage of the model group were significantly increased （P<0.01）， the level of IL-10 was significantly decreased （P<0.01）. Compared with the model group， the levels of TNF-α and IL-1β in synovium were decreased in all administration groups （P<0.05）， but the difference of the levels of MMP-13 and IL-10 in synovium had no statistical significance. The level of inflammatory mediators in cartilage was not affected in all administration groups. Compared with the sham operation group， the levels of TNF-α and IL-β in serum of the model group were significantly increased （P<0.01）， the level of IL-10 was decreased （P<0.05）. Compared with the model group， the level of TNF-α in the high-dose FHDP group was decreased （P<0.05）， and the level of IL-10 was increased in all administration groups （P<0.05， P<0.01）. The difference of the level of IL-β in all administration groups had no statistical significance. ConclusionFHDP attenuated the synovitis of KOA rats. FHDP exert the effect on the releasing of proinflammatory cytokines and MMP by inhibiting the polarization of M1 macrophages in synovium， and had no significant effect on the polarization of M2 macrophages. Modulating the imbalanced polarization of synovial macrophages was a possible mechanism of FHDP on attenuating synovitis and treating KOA.

Therapeutic Effect of Huangqi Chifengtang on Cerebral Infarction Rat Model and Its Effect on Expression of ZO-1， Claudin-5， P-gp and MRP1 Protein in Brain Tissue / 中国实验方剂学杂志

ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Chifengtang on middle cerebral artery embolism（MCAO） rat model. MethodThe 90 SPF male rats were randomly divided into sham operation group， model group， high， medium and low dose groups of Huangqi Chifengtang （8.10，4.05，2.025 g·kg-1） and positive drug group （Naoxintong 0.32 g·kg-1）. MCAO rat model was established and intervened with Huangqi Chifengtang， and the neurological scores of each group were scored. The area of cerebral infarction was calculated by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Serum interleukin-6 （IL-6） and interleukin-1β（IL-1β） were detected by enzyme-linked immunosorbent assay （ELISA），The contents of matrix metalloproteinase-9（MMP-9）， vascular endothelial growth factor（VEGF） and vascular endothelial growth factor receptor 2 （VEGFR2）， the pathological changes of brain tissue in each group were observed by hematoxylin eosin （HE） staining. Western blot was used to detect zonule atresia protein-1（ZO-1）， tight junction protein-5（Claudin-5） and P-glycoprotein （P-gp） and multidrug resistance protein 1（MRP1）. ResultCompared with the sham operation group， the neurological function score and cerebral infarction rate of the model group were significantly increased（P<0.01）， and the levels of IL-6， IL-1β and MMP-9 in serum were significantly increased（P<0.01）， the levels of ZO-1 and Claudin-5 protein expression decreased significantly（P<0.01）， and the levels of P-gp and MRP1 protein expression increased significantly（P<0.01）. Compared with the model group， the neurological function score of rats in each administration group decreased significantly at 14 days （P<0.05，P<0.01）， the pathological changes of brain tissue effectively improved， the rate of cerebral infarction significantly reduced （P<0.01）， and the expression level of IL-6， IL-1β and MMP-9 in serum decreased significantly （P<0.05，P<0.01）， the content of VEGFR2 increased significantly （P<0.01）， and the content of VEGF increased significantly in high， medium dose and positive drug groups （P<0.05，P<0.01）. Although it decreased in low dose group， there was no significant difference. The levels of ZO-1 and Claudin-5 protein expression in brain tissue of high dose group and positive drug group increased significantly （P<0.05，P<0.01）， the level of MRP1 and P-gp protein expression decreased significantly （P<0.05）. ConclusionHuangqi Chifengtang can play a therapeutic role in rats with cerebral infarction by improving the pathological changes of brain tissue， reducing inflammatory reaction， promote angiogenesis and regulating the function of blood-brain barrier（BBB）.

Chemical Composition Analysis and Characterization of Reference Sample of Huangqi Guizhi Wuwutang Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

ObjectiveTo analyze the chemical composition of the reference sample of Huangqi Guizhi Wuwutang （lyophilized powder）， and to provide quality markers for the formulation of quality standards of this formula. MethodUltra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS） was performed on a Waters ACQUITY UPLC™ HSS T3 column （2.1 mm×100 mm， 1.8 μm）， the mobile phase was methanol （A） -0.1% formic acid aqueous solution （B） for gradient elution （0-8 min， 1%-20%A； 8-10 min， 20%-30%A； 10-12 min， 30%-35%A； 12-14 min， 35%-40%A， 14-23 min， 40%-55%A， 23-27 min， 55%-99%A； 27-28 min， 99%A； 28-28.5 min， 99%-1%A； 28.5-30 min， 1%A）， the column temperature was 40 ℃， the injection volume was 2 μL， and the flow rate was 0.3 mL·min-1. The mass spectrometry data of the reference sample of Huangqi Guizhi Wuwutang （lyophilized powder） were collected under positive and negative ion modes. The conditions of mass spectrometry were electrospray ionization （ESI）， scanning range of m/z 50-1 200， and impact energy of 10-30 eV. UNIFI 1.8 and Progenesis QI 2.0 software were used to analyze and characterize the chemical constituents in reference sample of Huangqi Guizhi Wuwutang （lyophilized powder） combined with reference comparison and literature review. ResultA total of 123 chemical constituents were identified， including 33 flavonoids， 26 glycosides， 18 organic acids， 11 terpenoids， 7 phenylpropanoids， 4 gingerol， 3 alkaloids， 3 amino acids， 2 amides and 16 other compounds. ConclusionThe established method can quickly and accurately characterize the chemical components in the reference sample of Huangqi Guizhi Wuwutang （lyophilized powder）， which can provide a basis for the selection of quality evaluation indicators of this formula， and provide a reference for its preparation research.

Therapeutic Mechanism of Huangqi Guizhi Wuwutang on Rheumatoid Arthritis / 中国实验方剂学杂志

ObjectiveTo reveal the mechanism of action of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis by pharmacological research based on its clinical application. MethodThe collagen-induced arthritis （CIA） rat model was established by injecting bovine type Ⅱ collagen and Freund's adjuvant at the tail， and was treated with different concentrations of Huangqi Guizhi Wuwutang. The rats were randomly divided into blank group， model group， methotrexate （0.9 mg·kg-1） group， and Huangqi Guizhi Wuwutang low- and high-dose （5.13， 20.52 g·kg-1·d-1） groups， with continuous intragastric administration for 4 weeks. The degree of joint swelling， weight， degree of foot swelling and arthritis index score were determined and the pathological changes of ankle joints were detected by hematoxylin and eosin （HE） staining to observe the therapeutic effect of Huangqi Guizhi Wuwutang on rheumatoid arthritis. In addition， enzyme-linked immunosorbent assay （ELISA） and Western blot were used to measure the expression of interleukin 1β （IL-1β）， interleukin 6 （IL-6）， interleukin 10 （IL-10） and tumor necrosis factor-α （TNF-α） in serum and the expression of nuclear factor kappa-B （NF-κB） pathway related proteins in synovial tissue， respectively to clarify the molecular mechanism of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis. ResultCompared with the conditions in blank group， the body weight and IL-10 level were decreased （P<0.01）， and the degree of foot swelling and arthritis index score， the levels of IL-1β， IL-6 and TNF-α， and the expression of NF-κB pathway related proteins were increased （P<0.01，） in the model group， with impaired morphology and function of the ankle joint. Additionally， compared with the model group， Huangqi Guizhi Wuwutang low- and high-dose groups had increased body weight of rats and IL-10 level （P<0.01）， and reduced degree of foot swelling and arthritis index score （P<0.05， P<0.01）， levels of IL-1β， IL-6 and TNF-α （P<0.01） and expression of NF-κB pathway related proteins （P<0.05， P<0.01）， with improved function and morphology of the ankle joint. ConclusionHuangqi Guizhi Wuwutang can significantly alleviate joint inflammatory injury by down-regulating NF-κB pathway and reducing the inflammatory response in CIA rats.

Effects of music relaxation training combined with Huangqi Jianzhong Decoction oral administration and foot bathing on fatigue status and quality of life in patients with cancer related fatigue / 国际中医中药杂志

Objective:To explore the effect of music relaxation training combined with Huangqi Jianzhong Decoction oral administration and foot bath on fatigue status and quality of life in patients with cancer related fatigue.Methods:A total of 90 patients with cancer related fatigue in our hospital from January 2019 to October 2020 were selected. The patients were divided into 2 groups by random number table method, with 45 in each group. Both groups were given health and diet education. The control group was given music relaxation training, and the observation group took Huangqi Jianzhong Decoction oral administration and foot bathing on the basis of the control group. Both groups have a course of treatment of 4 weeks, a total of 3 courses. The TCM syndrome scores were observed before and after treatment, and the Piper Fatigue Score (PFS) was used to evaluate fatigue status, and the Karnofsky functional state score was used to evaluate the quality of life of patients, evaluation of clinical efficacy.Results:After treatment, 43 patients in the observation group and 42 patients in the control group were analyzed. The total effective rate of the observation group was 83.72% (36/43), which was significantly higher than 64.29% (27/42) of the control group ( χ2=4.18, P=0.041). After treatment, the TCM syndrome scores in the observation group was significantly lower than that of the control group ( t=2.48, P=0.015). After treatment, the Karnofsky functional state score in the observation group was significantly higher than that of the control group ( t=2.83, P=0.006). After treatment, the emotion score (4.35±1.02 vs. 5.03±1.01, t=3.09), feeling score (4.31±1.06 vs. 5.00±1.02, t=3.06) and total score (4.23±1.07 vs. 4.87±1.10, t=2.72) in the observation group were significantly lower than those in the control group ( P<0.01). Conclusion:Music relaxation training combined with Huangqi Jianzhong Decoction oral administration and foot bathing can alleviate the fatigue state of CRF patients and improve the quality of life.

Effect of Modified Huangqi Gancaotang on Proliferation， Apoptosis， Invasion， Migration， and Epithelial-mesenchymal Transition of Non-small Cell Lung Cancer Cells Based on Wnt/β-catenin Pathway / 中国实验方剂学杂志

ObjectiveTo investigate effect of lyophilized powder of modified Huangqi Gancaotang on proliferation， apoptosis， invasion， migration， and epithelial-mesenchymal transition of human non-small cell lung cancer cells （A549， PC9） and possible mechanism. MethodEffect of 1.0， 2.0， 4.0， 8.0， 12.0 g·L-1 modified Huangqi Gancaotang on the proliferation of non-small cell lung cancer cells was detected by cell counting kit-8 （CCK-8） assay. A549 and PC9 cells were classified into the blank group and the low-， medium-， and high-dose Huangqi Gancaotang groups （2.0， 4.0， 8.0 g·L-1）. Plate cloning assay was used to examine the effect of modified Huangqi Gancaotang on cell cloning ability. Hoechst 33342 staining and flow cytometry were employed to detect the apoptosis， and scratch assay and Transwell migration assay were applied to examine cell migration and invasion abilities， respectively. Mammosphere assay was used to examine the sphere-forming ability of tumor cells， and real-time polymerase chain reaction （Real-time PCR） to detect the mRNA expression of stemness-related molecules octamer-binding transcription factor 4 （Oct-4）， human sex-determining region Y-box 2 （Sox2）， and homeobox transcription factor （Nanog） to assess cancer stem cell activity. The protein expression of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated death promoter （Bad）， Bcl-2-associated X protein （Bax）， cleaved Caspase-3， Caspase-3， E-cadherin， N-cadherin， vimentin， matrix metalloproteinase-2 （MMP-2）， β-catenin， c-Myc， Cyclin D1， and zinc-finger transcription factor （Slug） was determined by Western blot. ResultThe proliferation ability of A549 and PC9 cells was significantly inhibited after 24 h and 48 h treatment with 1.0， 2.0， 4.0， 8.0， and 12.0 g·L-1 lyophilized powder of modified Huangqi Gancaotang compared with that in the blank group and the inhibition was dose- and time-dependent （P<0.05， P<0.01）. Compared with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang suppressed the cloning ability of A549 and PC9 cells （P< 0.05， P<0.01）， and high-dose modified Huangqi Gancaotang induced apoptosis of A549 and PC9 cells （P<0.01）. In comparison with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang inhibited the invasion and migration of A549 and PC9 cells （P<0.05， P<0.01）. Compared with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang significantly decreased volume of the microspheres of A549 cells and the mRNA expression of Oct-4， Sox2， and Nanog in A549 and PC9 cells （P<0.05， P<0.01）. Compared with the blank group， the medium- and high-dose modified Huangqi Gancaotang down-regulated the expression of the anti-apoptotic protein Bcl-2 （P<0.05， P<0.01）， up-regulated the expression of pro-apoptotic proteins Bad， Bax， and cleaved Caspase-3/Caspase-3 （P<0.05， P<0.01） in A549 and PC9 cells， decreased the expression of MMP-2， N-cadherin， and vimentin （P<0.05， P< 0.01）， and raised the E-cadherin expression （P<0.05， P<0.01）. Moreover， the medium-dose and high-dose modified Huangqi Gancaotang all reduced the expression of β-catenin， c-Myc， Cyclin D1， and Slug in A549 and PC9 cells （P<0.01）. ConclusionModified Huangqi Gancaotang can inhibit the proliferation， invasion， migration， activity of cancer stem cells， and epithelial-mesenchymal transition of human non-small cell lung cancer （A549， PC9） cells and induce apoptosis， and the mechanism is the likelihood that it regulates Wnt/β-catenin signaling pathway.