2022 Vol. 38, No. 6

Display Method:
Research Development and Strategies of "Wind Elimination" Method in the Treatment of Childhood Asthma in Three Stages
WANG Shou-chuan, LIN Li-li, FENG Lu
2022, 38(6): 467-475. doi: 10.14148/j.issn.1672-0482.2022.0467
Abstract:
This paper points out that the congenital endowment of children with asthma is different, forming the latent constitution of "latent-wind", combined with the invasion of external evil by harmful wind, leads to the pathogenesis of asthma. It is proposed that, the middle and later stage should be increased in addition to the traditional asthma attack and remission staging methods. The "wind elimination method" in the treatment of three-stage pediatric asthma: during the attack stage, expel external wind and "latent-wind" elimination, downregulate qi, and relieve asthma; treatment methods for the middle and later stages are promoting healthy qi and suppressing "latent-wind"; during the remission period, restores organ qi and resists external wind, tonifying the lung, spleen, and kidney. This paper focuses on how to treat the three-stage asthma syndrome in children using the theory of eliminating wind; to clarify the evolution characteristics and syndrome differentiation law of the "latent-wind" in asthma syndrome. This paper provides theoretical support and implementation strategies for TCM clinical diagnosis and treatment of pediatric asthma, combining modern clinical and mechanism research progress.
The Guideline for the Diagnosis and Treatment of Pediatric Asthma in Chinese Medicine (Revised)
ZHAO Xia, QING Yan-hong, WANG You-peng, WANG Meng-qing, XUE Zheng, CHEN Jian, SUN Li-ping, WU Li-qun, WU Zhen-qi, ZHAO Qiong, WAN Li-sheng, HE Li, YU Shun, LI Lan, DAI Qi-gang, YOU Yan-nan, CHEN Bai-lei
2022, 38(6): 476-482. doi: 10.14148/j.issn.1672-0482.2022.0476
Abstract:
Based on the completion of literature search and evaluation, the Guidelines for the diagnosis and treatment of pediatric asthma in Chinese medicine (revised) was developed through two rounds of expert questionnaires, expert validation sessions, and public opinion solicitation. This revision proposes the scope of application, normative references, terminology and definitions, diagnosis, staging, syndrome differentiation, treatment, as well as prevention and care, aiming to provide an important reference for the clinical practice, treatment standard and quality evaluation of Chinese medicine in children with asthma.
Effects of Guben Fangxiao Decoction on Airway Epithelial Tight Junction Protein and CCSP in Mice of Asthma in Remission Stage
JIANG Yue, ZHANG Yu-lin, DONG Ying-mei, YAN Hua, ZHAO Xia
2022, 38(6): 483-489. doi: 10.14148/j.issn.1672-0482.2022.0483
Abstract:
  OBJECTIVE  To observe the effects of Guben Fangxiao Decoction (GBFXD) on airway tight junction protein and CCSP, and to explore the possible mechanism of GBFXD on airway inflammation in remission stage of asthma.  METHODS  Mouse model of asthma in remission stage was established by sensitization and stimulation of OVA combined with respiratory syncytial virus (RSV) infection. Mice were randomLy divided into six groups (n=6): control group, model group, GBFXD low-dose group (GBFXD-L), GBFXD medium-dose group (GBFXD-M), and GBFXD high-dose group (GBFXD-H) and montelukast group.The mice were developed asthma model and given different concentrations of GBFXD (12, 24, 36 g·kg-1)and montelukast sodium (2.6 mg·kg-1) by gavage according to groups for 28 days. The mice were sacrificed and samples were obtained after the intervention. Hematoxylin-eosin (HE) staining was used to observe the pathological features of lung tissues in each group. The expression of CCSP in lung tissues and alveolar lavage fluid (BALF) was determined by ELISA. Western blot, qPCR and immunohistochemical assay were used to detect the expression of ZO-1 and other airway epithelial tight junction proteins in lung tissues of mice in each group.  RESULTS  Compared with the control group, the model mice in remission stage are mainly infiltrated by inflammatory cells in the lung tissue (P < 0.01), and the infiltration of inflammatory cells in the lung tissues of mice was alleviated after the treatment (P < 0.01). The protein expression levels of ZO-1, Occludin and Desmocollin in the model group were significantly lower than those in the normal group (P < 0.05, P < 0.01), immunohistochemical results showed that the expression of the above indexes was reduced and the proteins were loosely arranged in the model group. Meanwhile, the mRNA expression levels of ZO-1, ZO-2, Vinculin and Catenin in the model group significantly decreased (P < 0.05, P < 0.01). Compared with the normal group, the expression of CCSP in the model group significantly decreased in lung tissue and BALF (P < 0.01), while the expression of CCSP in BALF and lung tissue was significantly increased by the intervention of low and high doses of GBFXD (P < 0.05, P < 0.01).  CONCLUSION  GBFXD can repair the epithelial injury and restore the epithelial barrier function by up-regulating the expression of epithelial tight junction protein and CCSP, an indicator of epithelial cell permeability change, so as to reduce the impact of various external pathogenic factors on the airway and preventing and treating asthma.
Mechanism of Qingfei Oral Liquid Flavonoids in Improving Lung Epithelial Barrier Damage in RSV-Infected Mice
LING Xiao-ying, SUN Xun, LI Wei-feng, YUAN Bin
2022, 38(6): 490-495. doi: 10.14148/j.issn.1672-0482.2022.0490
Abstract:
  OBJECTIVE  To investigate the protective mechanism of flavonoids in Qingfei Oral Liquid(Fla) on pulmonary epithelial barrier damage in respiratory syncytial virus (RSV)-infected mice.  METHODS  BALB/c mice were randomly divided into normal group, model group, low- and high-dose groups of flavonoids in Fla(30, 60 mg·kg-1·d-1), positive drug ribavirin group (46 mg·kg-1·d-1). RSV-infected mice model was established. After continuous administration for 4 d, the pathological changes of lung tissue were observed by HE staining; ELISA was used to detect the contents of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α); Protein content in BALF and BALF/serum fluorescence ratio were used to evaluate lung epithelial permeability; Western blot was used to detect the protein expression of Claudin-1, Occludin, ZO-1, p38 mitogen-activated protein kinase (p38 MAPK), p-p38 MAPK, TLR4 and Myd88 in lung tissue.  RESULTS  Compared with the model group, the lung pathological damage in Fla group was alleviated (P < 0.05, P < 0.01), the contents of IL-1β and TNF-α in BALF decreased (P < 0.05, P < 0.01), the protein content in BALF and the ratio of BALF/serum fluorescence decreased (P < 0.05, P < 0.01), the levels of Claudin-1, Occludin and ZO-1 in the lung tissue increased (P < 0.05, P < 0.01), the protein expression of p-p38 MAPK/p38 MAPK, TLR4 and Myd88 decreased (P < 0.05, P < 0.01).  CONCLUSION  Fla might inhibit the TLR4/MyD88/p38 MAPK pathway, increase the tight junction protein, and reduce lung inflammation, thereby improving the lung epithelial barrier damage caused by RSV.
The Network Pharmacological Study and Experimental Verification of Huanglong Antitussive Granule in the Treatment of Asthma
WANG Xuan, QIAN Gui-ying, LING Xiao-ying, YAN Hua, WANG Shou-chuan, SHAN Jin-jun
2022, 38(6): 496-503. doi: 10.14148/j.issn.1672-0482.2022.0496
Abstract:
  OBJECTIVE  Combining network pharmacology methods and experimental verification to explore the target and potential mechanism of Huanglong Antitussive Granule in treating asthma.  METHODS  With the help of TCMSP, TCMID, Swiss Target Prediction and other databases, the chemical components and targets of Huanglong Antitussive Granule are searched. Through GeneCards, OMIM, Disgenet, Drugbank database to screen out the disease targets of asthma. Use Cytoscape software to construct a network diagram of "drug-component-target-disease" and the interaction relationship between potential targets, and predict the mechanism of action through enrichment analysis. Mice model of asthma was constructed, and the results of network pharmacology enrichment analysis were verified by pathological staining, Elisa detection, q-PCR and Western Blot.  RESULTS  A total of 175 active ingredients, 1 026 drug targets, 900 disease targets for asthma and 153 common drug-disease targets were obtained in Huanglong Antitussive Granule. A total of 1 967 entries were obtained by GO enrichment analysis, and KEGG pathway enrichment screened out a total of 84 signaling pathways including TNF signaling pathways. Animal experiments show that Huanglong Antitussive Granule can effectively improve airway inflammation in asthmatic mice, and inhibit the MAPK and NF-κB signaling pathways by affecting the TNF signaling pathway.  CONCLUSION  This study initially revealed the mechanism of action of Huanglong Antitussive Granule in treating asthma, providing a reference for clinical medication and follow-up research.
Study on the Hypoglycemic Effect and Mechanism of Qidan Fang on Diabetic Mice
FENG Yi-fan, WU Li-ping, ZHANG Yi-ying, QIAN Da-wei, DUAN Jin-ao, SONG Xiao-xiong, LYU Gao-hong, ZHU Yue, XU Hui-qin
2022, 38(6): 504-510. doi: 10.14148/j.issn.1672-0482.2022.0504
Abstract:
  OBJECTIVE  To investigate the hypoglycemic effect of Qidan Fang on diabetic mice and explore its relevant mechanism.  METHODS  The SPF ICR mice were intraperitoneally injected with 120 mg·kg-1 alloxan after fasting for 16 h. The blood glucose was measured after fasting for 12 h after 5 d. The blood glucose of 10~25 mmol·L-1 was defined as the successful establishment of the diabetic mouse model. The male diabetic ICR mice were divided into the model group, metformin group, low-, medium- and high-dose Qidan Fang groups (0.34, 0.67, 1.34 g·kg-1·d-1, respectively), and administrated for 30 d. Additionally, normal male ICR mice were selected as the blank group. Body mass and fasting blood glucose (FBG) were recorded once every 10 d, and a glucose tolerance test (OGTT) was performed after 30 d. Serum insulin level was detected by ELISA, and then pancreas and liver tissues were collected for HE staining to observe their pathological changes. The expression levels of IRS-1, PI3K, AKT and GLUT4 in the liver tissues were determined by Western blot and qPCR.  RESULTS  Compared with the model group, the level of FBG and glucose tolerance decreased significantly (P < 0.01), and the serum insulin level increased significantly (P < 0.01), in the medium- and high-dose Qidan Fang groups; The tissue structure of the pancreas and liver was significantly improved, islet morphology recovered to a certain extent, and liver tissue abnormalities and vascular congestion were rarer in the low-, medium- and high-dose Qidan Fang groups. High dose of Qidan Fang could significantly up-regulate the protein expression levels of p-IRS-1/IRS-1, p-PI3K/PI3K, p-AKT/AKT and GLUT4 in liver tissues (P < 0.01), while significantly up-regulate the mRNA levels of IRS-1, PI3Kp85α, AKT1 and GLUT4 in liver tissues (P < 0.01).  CONCLUSION  Qidan Fang has a hypoglycemic effect on diabetic mice, and its mechanism is relative to the regulation of IRS-1/PI3K/AKT signaling pathway.
Meranzin Hydrate Effects Atherosclerosis and Depressive Comorbidity in ApoE-/- Rats by Antidyslipidemic, Anti-Inflammatory and Regulate Intestinal Flora
WANG Zhe, SUN Bo, REN Ping, HUANG Xi
2022, 38(6): 511-519. doi: 10.14148/j.issn.1672-0482.2022.0511
Abstract:
  OBJECTIVE  Focus on the efficacy of Meranzin hydrate (MH) in the treatment of comorbid AS and depression, and dicuss the mechanism of MH.  METHODS  Sprague Dawley rats with chow diets were elected as the control group, and ApoE-/- rats were fed high-fat diets for 12 weeks to establish the AS multimorbidity with depression model. ApoE-/- rats were randomly divided into the model group and intervention groups (MH 3.5 mg·kg-1, MH 7 mg·kg-1, and simvastatin 7 mg·kg-1 groups). n=6, for each group.  RESULTS  Treatment of comorbid AS and depression with MH could significantly decrease lipid levels, plaque areas, plasma level of inflammatory factors and hippocampus inflammatory cytokines, particularly tumor necrosis factor-α, interleukin(IL)-1β, and IL-6. MH could also improve hippocampal 5-hydroxytryptamine levels as well as depressive-like behavior and regulate intestinal flora.  CONCLUSION  This study demonstrates that MH reduces the inflammatory response as well as lipid levels and improves the composition of intestinal flora, which plays an important role against AS multimorbidity with depression.
Effect of Arisaema Cum Bile on Brain Tissue Injury and Inflammation in Febrile Seizures Model Mice
ZENG Ping, YAN Yu-mei, YU Hong-li, WU Hao, TAO Xing-bao, XIE Yu-wei, CHENG Yan-qiu, WANG Cai-xia, WANG He-peng, CHEN Jin-song
2022, 38(6): 520-526. doi: 10.14148/j.issn.1672-0482.2022.0520
Abstract:
  OBJECTIVE  To study the efficacy of Arisaema Cum Bile on antagonizing brain tissue damage and inflammation in febrile seizures model mice.  METHODS  Mice model of febrile seizures was established by using dry yeast suspension and pentylenetetrazol solution, and the effects of low, medium and high dose groups of Arisaema Cum Bile water extract on rectal temperature, convulsion latency and convulsion duration of model mice were investigated. HE staining was used to observe the morphology of mice hippocampal neurons. ELISA was used to detect the levels of serum cAMP and PGE2. Western blot method was used to detect the protein expressions of COX-2, iNOS, GFAP and GABAAR in brain tissue. The qPCR method was used to detect inflammatory factors IL-1β, IL-6, TNF-α mRNA expression in hippocampus tissue.  RESULTS  Compared with the model group, the high-dose Arisaema Cum Bile group could reduce the rectal temperature (P < 0.05), prolong the convulsion latency (P < 0.05), shorten the duration of convulsions (P < 0.01), and up-regulate the expression of GABAAR protein (P < 0.05); At the same time, it decreased the levels of serum cAMP and PGE2 (P < 0.01), and improved the morphological changes of hippocampal neurons, down-regulated the expressions of COX-2, iNOS and GFAP protein in the brain tissue (P < 0.05), and decreased the expression levels of IL-1β, IL-6 and TNF-α mRNA in the hippocampus (P < 0.05).  CONCLUSION  Arisaema Cum Bile has a good antipyretic convulsant effect, and its mechanism may be related to antipyretic and antagonizing brain tissue inflammation.
Study on the Formulation and in vitro Release of Evodiamine-Loaded Ginger-Derived Extracellular Vesicle-Like Nanoparticles
ZHAO Meng, LIU Zhuo-ya, YU Jia-min, WANG Rui, FAN Ming-jie, QIAO Hong-zhi
2022, 38(6): 527-533. doi: 10.14148/j.issn.1672-0482.2022.0527
Abstract:
  OBJECTIVE  Lipids from ginger-derived extracellular vesicle-like nanoparticles (EVNs) were extracted and used to prepare liposomes containing evodiamine (EVO) to improve its druggability.  METHODS  EVNs from ginger were separated by differential centrifugation, and the lipid extraction solvent was screened. Liposomes loaded with EVO (EVO@Lipo) were prepared by thin film dispersion method. The formulation and preparation process of liposome were optimized by orthogonal test with encapsulation rate as evaluation index. EVO@Lipo was characterized by particle size and potential analysis, differential calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR), and the drug release behavior of EVO@Lipo was investigated in vitro.  RESULTS  Trichloromethane, methanol-trichloromethane and ethanol-dichloromethane were screened as lipid extraction solvents. The optimized preparation conditions were methanol-trichloromethane (2∶1) as lipid extraction solvent, drug to lipid ratio of 1∶50, ultrasonic conditions of 60 W, 15 min. The encapsulation efficiency of EVO@Lipo was 88.21%, the average particle size was 194.9 nm, the PDI was 0.22, and the Zeta potential was -35.3 mV. Accumulated evidence suggests that EVO@Lipo is not a physical mixture of drugs and lipids. In vitro release experiments showed that EVO@Lipo could delay drug release.  CONCLUSION  Lipids from ginger EVNs can be used to load hydrophobic drug EVO, improve its solubility, and have a certain sustained release effect.
Preparation and in vitro Evalution of Glycyrrhizic Acid Self-Assembled Micelles Loading Tanshinone ⅡA
LIANG Qi-fan, CUI Ji-wei, ZHANG Xin-ru, GUO Yu-miao, WANG Ruo-ning, DI Liu-qing
2022, 38(6): 534-540. doi: 10.14148/j.issn.1672-0482.2022.0534
Abstract:
  OBJECTIVE  Encapsulate tanshinone ⅡA (Tan ⅡA) in glycyrrhizic acid (GL) self-assembled micelles, to investigate the preparation process and characterization, and evaluate the anti-glioma cell activity in vitro.  METHODS  Thin film dispersion method was used to prepare self-assembled micelles loaded with tanshinone ⅡA glycyrrhizic acid (TGM, Tan ⅡA-GL/Micelle). The particle size, polydispersity index (PDI) and Zeta potential of TGM were measured. The typical morphological characteristics of TGM were observed by transmission electron microscopy (TEM). HPLC was employed to determine the encapsulation rate of TGM, then the drug release of TGM under different pH conditions was investigated by dialysis bag method. The in vitro blood-brain barrier (BBB)-crossing ability of TGM was inspected by inverted fluorescence microscopy. Quantitative cell uptake experiment to investigate the specific mechanism of cell uptake. The cytotoxicity of free Tan ⅡA, GL and TGM on GL261 cells was investigated by MTT assay. The apoptotic effects of free Tan ⅡA, GL and TGM on GL261 cells were detected by flow cytometry.  RESULTS  Thin film dispersion method was the best method to prepare self-assembled micelles loaded with tanshinone ⅡA glycyrrhizic acid. The particle size of TGM was about (121.87±5.85) nm, and the encapsulation efficiency of Tan ⅡA was (88.54±14.27)%, which was almost spherical in shape and the preparation was uniform and stable. TGM was capable of effective BBB-penetrating and the mechanism of uptake of self-assembled micelles by glioma cells was the uptake of preparations by clathrin-mediated endocytosis. TGM enhanced GL261 cell apoptosis from 12.28% (Free Tan ⅡA) and 13.32% (Free GL) to 31.16% (TGM), providing a promising potential to the GBM treatment.  CONCLUSION  The TGM nano-micelles are self-assembled from GL and Tan IIA, which effectively overcomes low bioavailability and poor water solubility of Tan IIA. TGM can effectively cross the blood-brain barrier and be taken up by glioma cells through clathrin-mediated endocytosis, with good cytotoxicity and pro-apoptotic ability.
Study on the Difference of Saponins Profile of Platycodonis Radix from the Northern and Southern Regions Based on Liquid Chromatography-Mass Spectrometry
XU Wei-chen, LUO Zi-chen, HE Yu, DI Liu-qing, GUO Qing, SHAN Jin-jun
2022, 38(6): 541-549. doi: 10.14148/j.issn.1672-0482.2022.0541
Abstract:
  OBJECTIVE  To identify the spanions of 57 batches of Platycodonis Radix from 9 provinces and cities, draw and compare the saponins profile between southern and northern platycodons.  METHODS  UPLC-LTQ-Orbitrap-MS and LC-MS/MS were used to determine the saponins in Platycodonis Radix. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze data.  RESULTS  A total of 48 saponins were identified in Platycodonis Radix. Both PCA and OPLS-DA showed that the northern and southern samples were well distinguished, 23 different saponins were found. Target quantitative results showed that the contents of platycodin D, deapio-platycodin D, polygalacin D and deapio-platycodin D2 of southern samples were significantly higher than those of northern samples.  CONCLUSION  There are significant differences in saponins between the northern and southern Platycodonis Radix, and the target quantification method is fast and stable, which can provide references for the quality control of Platycodonis Radix.
Syndrome Differentiation and Treatment of Ulcerative Colitis from the Theory of Potential Toxin
LONG Dan, ZHU Ying
2022, 38(6): 550-553. doi: 10.14148/j.issn.1672-0482.2022.0550
Abstract:
The study summarizes the clinical experience of Professor Zhu Ying, the inherited instructor of academic experience from the national veteran Chinese medicine experts, in the syndrome differentiation and treatment of ulcerative colitis from the theory of potential toxin. It is believed that the core pathogenesis of ulcerative colitis includes vital qi deficiencies leading to lingering pathogens, and collateral damages caused by the potential toxin. In view of that, we propose the treatment principles of treating and differentiating both syndromes and root causes by stages, controlling the development of existing disease by expelling the potential toxin, as well as preventing the recurrence of the disease by caring the spleen and kidney. Clinically, the treatment is to support healthy energy by fortifying the spleen, supplemented by draining the liver and warming the kidneys. In addition, some therapies such as eliminating wind, purging heat, resolving dampness, dispersing pain, and expelling phlegm are applied to cleanse the potential toxin.
A Study of Ancient Literature on Classical Formula Houpu Mahuang Decoction
ZHOU Li-ping, SUN Yu-bo, FAN Xin-sheng
2022, 38(6): 554-558. doi: 10.14148/j.issn.1672-0482.2022.0554
Abstract:
Houpu Mahuang decoction, from the Essentials from the golden cabinet, is one of the classical formulas included in the Catalogue of ancient classical formulas (first batch). Therefore, it is important to sort out and analyze the key information of the composition of this formula. The key information was sorted out through collecting and organizing the ancient medical books containing Houpu Mahuang decoction in ancient times, as well as analyzing the efficacy, indications, dosage and decoction method of the medicines in the formula. The medicines in the formula were examined with the use of scales, measuring vessels and the comparing objects, to provide reference for clinical application and the development of the formula preparation.