Mechanism of Yupingfeng San Against Allergic Rhinitis Through ROS/NLRP3/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the therapeutic effect of Yupingfeng San on allergic rhinitis （AR） and its effect on Reactive oxygen species （ROS）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3）/cysteinyl aspartate specific proteinase-1 （Caspase-1） pathway. MethodSPF mice were randomly divided into control group， model group， loratadine group （0.9 mg·kg-1）， and low， medium， and high dose Yupingfeng San groups （6， 12， 24 mg·kg-1）， with 10 mice in each group. The control group was given routine feeding， and the other groups were intraperitoneally injected with ［ovalbumin（OVA） + Al（OH）₃ + phosphate buffer solution（PBS）］ suspension once every other day for seven consecutive times. After seven days， 10% OVA solution was instilled in the nose， two times each day for seven consecutive days. After successful modeling， each administration group was intraperitoneally injected with different doses of Yupingfeng San Decoction， and the control group and model group were intraperitoneally injected with an equal volume of normal saline. Symptoms of sneezing， scratching， and runny nose were recorded and scored daily. The levels of ovalbumin specific immunoglobulin E （OVA-sIgE）， histamine， eosinophil cationic protein （ECP）， prostaglandin D2 （PGD2）， interleukin-1β （IL-1β）， interleukin-18 （IL-18）， interleukin-4 （IL-4）， and γ interferon （IFN-γ） in nasal lavage solution and serum of mice were detected by enzyme-linked immunosorbent assay （ELISA）. The damage status of the nasal mucosa was observed by hematoxylin-eosin （HE） staining. The number of goblet cells in the nasal mucosa of mice was observed by periodic acid-Schiff （PAS） staining. The expression of NLRP3 protein in the nasal mucosa of mice was detected by immunohistochemistry. Western blot was used to detect the expressions of NLRP3， cleaved Caspase-1， and cleaved gasdermin D （GSDMD） proteins in the nasal mucosa. The test kit was used to detect the changes in ROS in the nasal cavity of mice in each group. ResultCompared with the control group， the nasal symptoms of the model group were significantly aggravated， and the levels of inflammatory factors OVA-sIgE， histamine， ECP， PGD2， IL-1β， IL-18， and IL-4 in serum and nasal lavage solution were increased （P<0.05，P<0.01）. The levels of IFN-γ were decreased （P<0.05，P<0.01）. The histopathological score， goblet cell number， and ROS content were significantly increased （P<0.01）， and the expressions of pyrodeath-related proteins NLRP3， cleaved Caspase-1， and cleaved GSDMD were increased （P<0.01）. Compared with the model group， the nasal symptoms of the loratadine group and Yupingfeng San groups were significantly relieved， and the levels of inflammatory factors OVA-sIgE， histamine， ECP， PGD2， IL-1β， IL-18， and IL-4 in serum and nasal lavage solution were decreased （P<0.05，P<0.01）. The levels of IFN-γ were increased （P<0.05，P<0.01）. The histopathological scores， goblet cell number， and ROS content were significantly decreased （P<0.05，P<0.01）， and the expressions of pyrodeath-related proteins NLRP3， cleaved Caspase-1， and cleaved GSDMD were decreased （P<0.05，P<0.01）. Compared with the loratadine group， the curative effect of the high dose Yupingfeng San group was further increased （P<0.05，P<0.01）. ConclusionYupingfeng San has a therapeutic effect on AR， and its specific effect may be related to the inhibition of ROS/NLRP3/Caspase-1-induced cell pyroptosis.

Yiqi Wenyang Huwei Decoction Regulates PI3K/Akt/mTOR Pathway to Treat Bronchial Asthma in Rats / 中国实验方剂学杂志

ObjectiveTo observe the regulatory effects of Yiqi Wenyang Huwei decoction （YWHD） on autophagy and phosphatidylinositol 3-kinase （PI3K）/protein kinase B（Akt）/mammalian target of rapamycin （mTOR） signaling pathway in asthmatic rats and bronchial epithelial cells （16HBE） and further reveal the mechanism of YWHD in treating bronchial asthma （BA）. MethodForty-eight rats were randomly assigned into normal group， model group， dexamethasone group， and low-， medium-， and high-dose YWHD groups， with 8 rats in each group. The rat model of BA was established by intraperitoneal injection with ovalbumin （OVA） + aluminum hydroxide suspension and atomizing inhalation with OVA for 2 weeks. The normal group was administrated with an equal dose of normal saline. The bronchial maximum airway resistance （Max Rrs） induced by methacholine chloride （Mch） was determined by an animal lung function evaluation system. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of interleukin （IL）-4， IL-13， IL-6， IL-33， IL-25， tumor necrosis factor-α （TNF-α）， and immunoglobulin E （IgE） in the bronchial alveolar lavage fluid. Hematoxylin-eosin （HE） and Masson staining were used for observation of the pathological changes of bronchi in the lung tissue. The immunofluorescence assay was employed to measure the levels of the autophagy-associated proteins LC3B and Beclin1. The IL-13-induced autophagy of 16HBE cells exposed to the YWHD-containing serum was observed， and the autophagy level was traced by mRFP-GFP-LC3 adenovirus infection. The protein levels of LC3Ⅱ/Ⅰ， p-PI3K， p-Akt and p-mTOR were determined by Western blot. ResultCompared with the normal group， the model group showed increased Max Rrs （P<0.01） and elevated levels of IL-4， IL-13， IL-6， IL-33， IL-25， TNF-α， and IgE in the bronchial alveolar lavage fluid （P<0.05，P<0.01）. The modeling caused focal infiltration of inflammatory cells and lymphocytes around bronchus and blood vessels， epithelial goblet cell metaplasia， and increased subepithelial collagen deposition. Furthermore， it up-regulated the protein levels of LC3B and Beclin1 （P<0.01）， promoted the autophagy flux of GFP to mRFP in 16HBE cells induced by IL-13， down-regulated the protein levels of p-PI3K， p-Akt， and p-mTOR， and increased the LC3Ⅱ/Ⅰ ratio （P<0.01）. Compared with the model group， medium- and high-dose YWHD decreased Max Rrs （P<0.01）， lowered the levels of IL-4， IL-13， IL-6， IL-33， IL-25， TNF-α， and IgE in the bronchial alveolar lavage fluid （P<0.05， P<0.01）， and reduced lymphocyte and granulocyte infiltration in bronchi of the lung tissue， epithelial goblet cell metaplasia， and subepithelial collagen deposition. Moreover， they down-regulated the protein levels of LC3B and Beclin1 （P<0.05， P<0.01）， decreased the autophagy flux of GFP to mRFP， up-regulated the protein levels of p-PI3K， p-Ak， and p-mTOR， and decreased the LC3Ⅱ/Ⅰ ratio （P<0.05， P<0.01）. ConclusionYWHD ameliorates airway hyperresponsiveness and airway inflammation and inhibits the autophagy of airway epithelial cells in the lung tissue of BA rats by activating the PI3K/Akt/mTOR signaling pathway.

Effects of Yupingfeng granules on tumor metastasis of Lewis tumor-bearing mice by adjusting the EMT / 中国药房

OBJECTIVE To explore the effects of Yupingfeng granules on the improvement of epithelial barrier function and the inhibition of tumor metastasis by regulating epithelial-mesenchymal transition （EMT）. METHODS Thirty C57BL/6 mice were randomly divided into the normal group （distilled water），model group （distilled water） and Yupingfeng granule group （40 g/kg）， with 10 mice in each group. The model group and Yupingfeng granule group were inoculated with Lewis lung cancer cells subcutaneously in the right armpit to induce the spontaneous lung metastasis model. After modeling，each group was given water/ relevant medicine intragastrically，once a day，for 15 consecutive days. The effects of Yupingfeng granules on tumor metastasis were investigated by observing or determining the pathomorphology of lung tissue，metastatic lesion count on the surface of the lung， tumor metastatic lesion and the expression of carcinoembryonic antigens. qRT-PCR and Western blot assay were used to detect the mRNA and protein expressions of β-catenin，E-cadherin and vimentin in the lung tissue of mice. RESULTS Compared with the model group，the total number of pulmonary metastases on the surface was decreased significantly in Yupingfeng granule group （P＜ 0.05），the general morphology of lung tissue was recovered，and the expression of carcinoembryonic antigen in lung tissue was significantly decreased （P＜0.05）. mRNA and protein expressions of E-cadherin in lung tissue were significantly increased （P＜ 0.05），while mRNA and protein expressions of vimentin and β-catenin were significantly decreased （P＜0.05）. CONCLUSIONS Yupingfeng granules can inhibit EMT by regulating the expression of β-catenin，thus improving epithelial barrier function，and inhibiting the ability of tumor cells to invade and metastasize.

Effect of Yupingfeng Granules （玉屏风颗粒） on Metabonomics of Alveolar Lavage Fluid and Gut Microflora in Chronic Obstructive Pulmonary Disease Model Rats / 中医杂志

ObjectiveTo explore the possible mechanism of Yupingfeng Granules （玉屏风散） in preventing and treating chronic obstructive pulmonary disease （COPD） from the perspective of “lung-gut axis”. MethodsThirty-two male Wistar rats were randomly divided into normal group，model group， roxithromycin group and Yupingfeng Granules group， with 8 rats in each group. Except for the normal group， the rat model of COPD was prepared by intratracheal instillation of lipopolysaccharide （LPS） combined with smoking for 12 weeks. Since the fifth week of modeling，the roxithromycin group and the Yupingfeng Granules group were given 31.5 mg/（kg·d） and 1.575 g/（kg·d） of corresponding drugs respectively by gavage，and normal group and model group were given 10 ml/（kg·d） physiolo-gical saline. Sample was collected 24 hours after the last administration. The pathological changes of lung tissue were observed using HE staining； Ultrahigh performance liquid chromatography quadrupole time of flight mass spectrometry （UHPLC-QTOFMS） was used to detect the differential metabolites in alveolar lavage fluid （BALF） in all groups but roxithromycin group；16S rDNA sequencing technology was used to detect the changes of intestinal flora， and the association analysis was conducted between the differential metabolites and the differential flora. ResultsCompared with the normal group， the model group showed an increase in goblet cells in the small bronchial wall， disappearance of the smooth muscle layer of the bronchial wall， and infiltration of inflammatory cells； compared with the model group， roxithromycin group showed slight alveolar interstital edema， and obviously reduced inflammatory cell， while no obvious alveolar interstital edema was observed in the Yupingfeng Granules group， showing a small amout of inflammatory cell infiltration. The results of the BALF differential metabolite screening showed that compared with the normal group， 12 substances were upregulated and 19 substances were downregulated in the model group； compared with the model group， 37 substances in the Yupingfeng Granules group were upregulated and 43 substances were downregulated KEGG analysis yielded a total of 2 metabolic pathways， glycerophospholipid metabolism， and unsaturated fatty acid biosynthetic metabolism； compared with the model group， choline， acetylcholine， glycerol-3-phosphate， glycerophosphate choline， palmitic acid， and arachidonic acid showed an upward trend， while stearic acid and docosahexaenoic acid showed a downward trend in Yupingfeng Granules group （P＜0.05）. The results of the intestinal flora showed that， there are 80 different species between the normal group and the model group， and 65 different species between the model group and Yupingfeng Granules group. Among the top 5 species with relative abundance levels，compared with the model group， the level of Prevotella_9，Ruminococcaceae_UCG-005，Ruminiclostridium_6 increase，and Lactobacillus，Bacteroides decrease（P＜0.05）.The results of the correlation analysis showed that， in the normal and model groups， arachidonic acid was negatively correlated with Oribacterium（r=-0.753，P＜0.01）； in the Yupingfeng Granules group and model group， stearic acid and Bacteroides（r=0.788）， Mycobacterium（r=0.826），［Eubacterium］_Ruminantium_Group（r=0.770） was positively correlated（P＜0.01）， Arachidic acid was negatively correlated with Roseiarcus（r=-0.779）， glycerol-3-phosphate was negatively correlated with Desulfovibrio（r=-0.758）， Arachidonic acid was negatively correlated with Oribacterium（r=-0.753）， and Palmitic acid was negatively correlated with Pseudolabs（r=-0.750，P＜0.01）. ConclusionYupingfeng Granules can affect the metabolism of BALF and the flora structure of intestinal microorganisms， and regulating the balance of “lung-gut axis” may be one of the mechanisms of Yupingfeng Granules in treatment of COPD.

Efficacy of Yupingfeng granule combined with cetirizine in the treatment of allergic rhinitis and its effects on serum inflammatory factor level / 中国基层医药

Objective:To investigate the efficacy of Yupingfeng granule combined with cetirizine in the treatment of allergic rhinitis and its effects on serum inflammatory factor level. Methods:A total of 162 patients with allergic rhinitis admitted to Zhejiang Provincial Hospital of Chinese Medicine from January 2021 to March 2022 were included in this prospective controlled study. They were randomly divided into a control group and an observation group ( n = 81/group). The control group was treated with cetirizine and the observation group was treated with Yupingfeng granule combined with cetirizine. All patients were treated for 4 weeks. Clinical efficacy was compared between the two groups after 4 weeks of treatment. Main symptom score, nasal function indexes (total nasal airway resistance, nasal minimal cross-sectional area, and 0-5 cm nasal cavity volume), levels of inflammatory factors (interleukin-4, interleukin-6, and interleukin-10), and Rhinoconjunctivitis Quality of Life Questionnaire scores were compared between the two groups before and after 4 weeks of treatment. Results:Total response rate in the observation group was significantly higher than that in the control group [92.59% (75/81) vs. 79.01% (64/81), χ2 = 6.13, P < 0.05]. After 4 weeks of treatment, the scores of nasal congestion, nasal itching, and sneezing in the observation group were (0.63 ± 0.20) points, (0.70 ± 0.21) points, and (0.54 ± 0.17) points, which were significantly lower than (1.07 ± 0.23) points, (1.08 ± 0.24) points, and (0.89 ± 0.22) points in the control group ( t = 12.99, 10.72, 11.33, all P < 0.05). After 4 weeks of treatment, total nasal airway resistance in the observation group was significantly lower than that in the control group [(0.17 ± 0.05) kPa·s -1·L -1vs. (0.26 ± 0.06) kPa·s -1·L -1, t = 10.37, P < 0.05]. Nasal minimal cross-sectional area and 0-5 cm nasal cavity volume in the observation group were (0.94 ± 0.17) cm 2 and (9.74 ± 0.89) cm 3, respectively, which were significantly higher than (0.76 ± 0.10) cm 2 and (8.43 ± 0.78) cm 3 in the control group ( t = 8.21, 9.96, both P < 0.05). After 4 weeks of treatment, serum levels of interleukin-4 and interleukin-6 in the observation group were (67.79 ± 9.94) ng/L and (6.74 ± 1.42) ng/L, respectively, which were significantly lower than (104.31 ± 14.45) ng/L and (10.29 ± 2.56) ng/L in the control group ( t = 18.74, 10.91, both P < 0.05). Serum level of interleukin-10 in the observation group was significantly higher than that in the control group [(17.97 ± 2.54) ng/L vs. (12.48 ± 2.46) ng/L, t = 13.97, P < 0.05]. After 4 weeks of treatment, Rhinoconjunctivitis Quality of Life Questionnaire score in the observation group was significantly lower than that in the control group [(27.43 ± 8.82) points vs. (38.95 ± 7.76) points, t = 8.82, P < 0.05). Conclusion:Yupingfeng granule combined with cetirizine is highly effective on allergic rhinitis. The combined therapy can reduce clinical symptoms and inflammatory reactions, improve nasal function, and thereby improve quality of life.

Study on protective effect and mechanism of Modified Yupingfeng nasal spray on nasal mucosal injury in AR model rats / 中国药房

OBJECTIVE To study protective effect and potential mechanism of Modified yupingfeng nasal spray （YPF+） on nasal mucosal injury in allergic rhinitis （AR） model rats. METHODS AR model was induced by ovalbumin （OVA） and randomly divided into model group， YPF+group （50 µg/side，twice a day）， positive control group （Mometasone furoate aqueous nasal spray， 50 µg/side，once a day）； the blank group was set up， with 10 rats in each group. Administration groups were given relevant medicine， and blank group and model group were given equivalent normal saline for consecutive 4 weeks. Thirty minutes after last medication， the behavioral scores of rats were recorded， and the pathological changes of their nasal mucosa tissue were observed. The level of reactive oxygen species （ROS） in nasal mucosa tissue was detected. The protein and mRNA expressions of nucleotide- binding oligomerization domain-like receptor protein 3 （NLRP3），caspase-1，gasdermin D （GSDMD） were detected； the contents of interleukin-1β （IL-1β） and IL-18 in serum were also determined. RESULTS Compared with blank group， in model group， the nasal mucosa tissue structure was disordered， inflammatory cells infiltrated seriously， and lamina propria vascular dilation was visible； its behavioral score and pathological score， the level of ROS， protein and mRNA expressions of NLRP3， caspase-1 and GSDMD， serum contents of IL-1β and IL-18 in nasal mucosa tissue were increased significantly （P＜0.05）. Compared with model group， the symptoms of nasal mucosal injury in rats of each drug group were improved to varying degrees， and the above indicators were significantly reduced （P＜0.05）. CONCLUSIONS YPF+ may improve nasal mucosal injury of rats， relieve AR symptoms such as sneezing， itchy nose， runny nose， the mechanism of which may be associated with reducing the production of ROS in nasal mucosa and downregulating NLRP3/caspase-1/ GSDMD pathway.

Therapeutic effect of Yupingfeng powder combined with extensively hydrolyzed milk formulas on milk protein allergy in infants / 中国基层医药

Objective:To investigate the efficacy of Yupingfeng powder combined with extensively hydrolyzed milk formulas in the treatment of milk protein allergy in infants and its effects on immune function, interleukin-10 (IL-10) and IL-22 levels. Methods:Eighty infants with milk protein allergy who received treatment in Hangzhou Children's Hospital from January 2020 to January 2021 were included in this study. They were randomly assigned to Yupingfeng powder and conventional treatment groups, with 40 infants per group. An additional 80 infants who concurrently received health examination in the same hospital were included in the control group. The conventional group was treated with extensively hydrolyzed milk formulas. The Yupingfeng powder group was treated with Yupingfeng powder combined with extensively hydrolyzed milk formulas. All infants were treated for 30 successive days. Clinical efficacy, serum total immunoglobulin (IgE), milk specific IgE (sIgE), peripheral blood CD 4+CD 25+ Treg, IL-10 and IL-22 levels were compared between groups. Results:Total response rate in the Yupingfeng powder group was significantly higher than that in the conventional treatment group [92.50% (37/40) vs. 72.50% (29/40), χ2 = 5.54, P < 0.05]. Serum total IgE and milk sIgE levels in the Yupingfeng powder group were (132.93 ± 14.61) IU/L and (0.62 ± 0.14) IU/L, respectively, which were significantly lower than (150.27 ± 16.22) IU/L and (0.85 ± 0.17) IU/L in the conventional treatment group ( t = 5.02, 6.61, both P < 0.05). The expression of CD 4+CD 25+ Treg in the Yupingfeng powder group was significantly higher than that in the conventional treatment group [(13.29 ± 1.40)% vs. (11.84 ± 1.27)%, t = 4.85, P < 0.05). CD 4+CD 25+ Treg expression in the Yupingfeng powder group was not significantly different from that in the control group [(13.40 ± 2.03)%, t = 0.31, P = 0.759]. IL-10 in the Yupingfeng powder group was significantly higher than that in the conventional treatment group [(34.57 ± 4.07) μg/L vs. (22.19 ± 2.15) μg/L, t = 17.01, P < 0.05]. IL-22 level in the Yupingfeng powder group was significantly lower than that in the conventional treatment group [(2.20 ± 0.42) ng/L vs. (5.28 ± 0.79) ng/L, t = 21.77, P < 0.05]. IL-10 and IL-22 levels in the Yupingfeng powder group were not different from those in the control group [(35.53 ± 3.85) μg/L, (2.13 ± 0.53) ng/L, t = 1.26, P = 0.209; t = 0.73, P = 0.468]. Conclusion:Yupingfeng powder combined with extensively hydrolyzed milk formulas is highly effective on milk protein allergy. The combined therapy can improve the immune function of infants, enhance IL-10 level, and decrease IL-22 level.

Identifying absorbable bioactive constituents of Yupingfeng Powder acting on COVID-19 through integration of UPLC-Q/TOF-MS and network pharmacology analysis / 中草药·英文版

Objective: Yupingfeng Powder (YPF), a kind of preventative patent medicine, is chosen for treatment of coronavirus disease 2019 (COVID-19) due to its high frequency application in respiratory tract diseases, such as chronic obstructive pulmonary disease, asthma, respiratory tract infections, and pneumonia, with the advantage of reducing the relapse rate and the severity. However, the active components of YPF and the mechanisms of components affecting COVID-19 are unclear. This study aimed to determine active constituents and elucidate its potential mechanisms. Methods: Ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q/TOF-MS) and liquid chromatography-triple quadrupole mass spectrometry (LC-QQQ-MS) were used to determine the components and absorbable constituents of YPF. Secondly, TCMSP, Drugbank, Swiss and PharmMapper were used to search the targets of absorbable bioactive constituents of YPF, and the targets of COVID-19 were identified based on GeneCards and OMIM databases. STRING database was used to filter the possible inter-protein interactions. Thirdly, Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis were performed to identify molecular function and systemic involvement of target genes. Results: A total of 61 components of YPF and 36 absorbable constituents were identified through UPLC-Q/TOF-MS. Wogonin, prim-O-glucosylcimifugin, 5-O-methylvisamminol, astragaloside IV and 5-O-methylvisamminol (hydroxylation) were vital constituents for the treatment of COVID-19, and RELA, TNF, IL-6, MAPK14 and MAPK8ere recognized as key targets of YPF. The major metabolic reactions of the absorbed constituents of YPF were demethylation, hydroxylation, sulfation and glucuronidation. GO and KEGG pathway analysis further showed that the most important functions of YPF were T cell activation, response to molecule of bacterial origin, cytokine receptor binding, receptor ligand activity, cytokine activity, IL-17 signaling pathway, Chagas disease, lipid and atherosclerosis, etc. Conclusion: The approach of combining UPLC-Q/TOF-MS with network pharmacology is an effective tool to identify potentially bioactive constituents of YPF and its key targets on treatment of COVID-19.

Effects and mechanism of Chinese medicine Jiawei Yupingfeng in a mouse model of allergic rhinitis / 中西医结合学报

OBJECTIVE@#Chinese medicine has the potential to modulate allergic rhinitis (AR). There have been studies investigating the treatment efficacy of Yupingfeng San, alone or in combination with other ingredients, in AR, though few have studied the potential mechanisms of these drugs. In the present study, we measured the effects of Jiawei Yupingfeng (JWYPF), a traditional Chinese medicine formula, on mice with ovalbumin-induced AR and explored its underlying mechanism of action.@*METHODS@#Forty BALB/c mice were randomly divided into normal control, allergy control and two treatment groups of ten mice each. In the normal control group, mice were sensitized and challenged with saline. The mice in the allergy control and treatment groups were sensitized and challenged with ovalbumin and aluminum hydroxide gel. The treatments of JWYPF and Nasonex were administered intranasally in the AR mice for one week. Several signs of allergic inflammation, such as nasal eosinophils and inflammatory cytokines, were measured to determine the underlying mechanisms.@*RESULTS@#Mice in the JWYPF and Nasonex groups had significantly lower AR symptom scores than those in the allergy control group (the mean differences between JWYPF and the allergy control, and Nasonex and the allergy control were -2.00 ± 0.35 and -2.40 ± 0.32). After treatment with JWYPF and Nasonex, the levels of ovalbumin-specific IgE and histamine were significantly reduced, as were the levels of interlukin-4 and transforming growth factor-β, while interferon-γ levels were increased (all P < 0.0001, vs. allergy control). These two treatments also significantly inhibited eosinophil and mast cell infiltration into the nasal cavity but were not statistically different from one-another.@*CONCLUSION@#JWYPF has a potential therapeutic effect on AR via adjusting the rebalance of T helper 1 and T helper 2.

Mechanism of Yupingfeng Granule on CU Rats Based on IL-23/IL-17 Inflammation Axis / 中国实验方剂学杂志

Objective::To study the effect of Yupingfeng granule on the degranulation of skin mast cells in chronic urticaria (CU) rats and the intervention mechanism of interleukin-23(IL-23), interleukin-17(IL-17) inflammation axis. Method::Totally 60 SPF SD rats were selected and randomly divided into normal group (normal saline), model group (normal saline), and loratadine group (0.9 mg·kg-1·d-1), high-dose Yupingfeng granules group (4.05 g·kg-1·d-1), middle-dose group (2.7 g·kg-1·d-1), low-dose group (1.35 g·kg-1·d-1). The CU rat model was reproduced through intraperitoneal injection of ovalbumin with aluminum hydroxide suspension and DTP vaccine. Histopathological changes of rat skin were observed by hematoxylin-eosin (HE) staining. Degranulation of mast cells in rat skin was determined by toluidine blue staining. IL-23 and IL-17 protein expressions in skin tissue were determined by immunohistochemistry (IHC). IL-23 and IL-17 mRNA transcription levels in skin tissue were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result::Yupingfeng granules can significantly alleviate the pathological manifestations of dermal edema, collagen beam distance, inflammatory cell infiltration of CU rats, and reduce the degranulation reaction of skin tissue mast cells in CU rats. The IL-23, IL-17 mRNA and protein expressions of the skin of model group were significantly increased compared with the normal group (P<0.05, P<0.01). Compared with the model group, Yupingfeng granules can significantly down-regulate IL-23 mRNA and protein expressions of CU rats (P<0.05, P<0.01). Yupingfeng granules had no significant regulatory effect on IL-17. Conclusion::Yupingfeng granule can significantly reduce the degranulation of mast cells in skin tissue of CU rats, and improve the pathological manifestations, such as dermal edema, serous exudation and inflammatory cell infiltration. The mechanism may be related to inhibiting the secretion of IL-23 pro-inflammatory cytokines and improving CU lesions.

Effect of Yupingfeng San on Expressions of GATA3，Foxp3 in Allergic Rhinitis Mice / 中国实验方剂学杂志

Objective:To study the effect of Yupingfeng San （YPFS） on the expressions of GATA binding protein 3 （GATA3） and forkhead transcription factor3 （Foxp3） in ovalbumin （OVA）-induced lung tissue of allergic rhinitis （AR） mice， and explore the mechanism of YPFS on AR. Method:The allergic rhinitismice model was established by intraperitoneally injecting with OVA and Al （OH）3， and challenged with OVA intranasally. The mice were divided into four groups： normal，model，chloretadine（3 mg·kg-1） and YPFS（6.5 g·kg-1） group， the corresponding drugs were orally administrated for three weeks. At the end of administration，the infiltration of inflammatory cells， such as mast cells， eosinophils and neutrophils in nasal mucosa， were observed by htoxylin eosin （HE） staining. The serum concentrations of OVA-specific IgE and cytokines ［interleukin-4（IL-4）， IL-5 and γ-interferon （INF-γ）］ were determined by enzyme-linked immune sorbent assay （ELISA）. The expressions of GATA3 and Foxp3 proteins in nasal mucosa tissue were detected by Western blot. Result:The AR mice had such symptoms as scratching， sneezing and running nose. Nasal mucosa section by HE staining showed significant desquamation of AR mouse nasal mucosa cilia， obvious tissue stromal edema， telangiectasia， and a large number of eosinophilic cells， lymphocytes， plasma cells infiltration. YPFS obviously improved nasal symptoms of allergic rhinitis mice. Nasal mucosa epithelial structure was complete and arranged evenly， with no obvious tissue clearance edema and vasodilation， and inflammatory cell infiltration was significantly reduced. Compared with normal group， the levels of OVA specific IgE， IL-4 and IL-5 in peripheral blood of AR model group were significantly higher（P<0.01）， and the INF-γ level was significantly lower （P<0.01）. Compared with AR model group， the administration of chloretadine and YPFS can significantly reduce the level of OVA specific IgE and IL-4， IL-5， and increase the level of INF-γ in AR mice peripheral blood （P<0.05， P<0.01）. Western blot results showed that compared with normal group， GATA3 protein expression was significantly increased， while Foxp3 protein expression was significantly decreased in AR model group （P<0.05， P<0.01）. Compared with AR model group， YPFS inhibited GATA3， and promoted Foxp3 protein expression （P<0.01）. Conclusion:YPFS has an effect in alleviating OVA-induced allergic rhinitis．YPFS may modulate the immune response by regulating the balance of Th2/Treg cells.

Study on active compounds of Yupingfeng San for prevention of coronavirus disease 2019 (COVID-19) based on network pharmacology and molecular docking / 中草药

Objective: To find out the active compounds of Yupingfeng San for the prevention and treatment of the coronavirus pneumoniadisease 2019 (COVID-19) using network pharmacology and molecular docking, with a purpose to find a better clinical use of Yupingfeng San. Methods: The effective ingredients and targets of Astragali Radix, Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix were searched from the traditional Chinese medicine system pharmacology analysis platform (TCMSP) website, and the protein and protein interactive network of Yupingfeng San was established using the String database (PPI). Cytoscape 3.6.1 software was used for data analysis to extract the Hub network from the PPI network. The KEGG pathway analysis was performed using the String database and the molecular docking was performed using ChemOffice, PyMOL, and Auto Dock software. Results: A total of 45 effective ingredients were obtained with limited screening conditions [oral bioavailability (OB) ≥ 30%; drug-like (DL) ≥ 0.18], and 345 potential targets and 15 key targets of Yupingfeng San were screened. A total of 50 pathways were obtained by KEGG pathway analysis, among which 25 main pathways were selected, including PIK3R1 target regulation PI3K-Akt signaling pathway, Ras signaling pathway, HIF-1 signaling pathway, and MAPK signaling pathways and T cell receptor signaling pathways. Conclusion: The active compounds in Yupingfeng San can inhibit the combination between SARS-CoV-2 protein and angiotensin-converting enzyme II (ACE2), thus regulating multiple signal pathways (PIK3R1, IGF1R, etc), which plays a role in the prevention of COVID-19.

Study of the effect of polysaccharides from Yupingfeng on keratinocytes / 药学实践杂志

Objective To study the immunomodulatory effect of Yupingfeng polysaccharides on keratinocytes in vitro.Methods The effects of Yupingfeng polysaccharides on the expression of various inflammatory factors in the activated keratinocytes were detected by the Real-time PCR method.The mRNA and protein expression levels of Filaggrin were detected by Real-time PCR and Elisa respectively.The western bolt assay was used to detect the regulation of NF-κB signaling pathway by polysaccharides.Results Yupingfeng polysaccharides inhibited the secretion of inflammatory factors in the activated keratinocytes, promoted the expression of Filaggrin and exhibited significant inhibitory effect on the NF-κB signaling pathway, which plays an important role in the inflammation and immune response.Conclusion Yupingfeng polysaccharides had an immune regulation effect on skin keratinocytes in vitro and alleviated the development of local inflammation.These results lay a theoretical foundation for the clinical application of Yupingfeng prescription.

Systematic evaluation and Meta-analysis on effectiveness and safety of Yupingfeng Granules on recurrent respiratory tract infection / 中国中药杂志

To evaluate the effectiveness and safety of Yupingfeng Granules in the treatment of recurrent respiratory tract infection.Six Chinese and English databases were retrieved,namely CNKI,Wan Fang,VIP,CBM,PubMed and Cochrane Library. Randomized controlled trials of Yupingfeng Granules in the treatment of recurrent respiratory tract infection were collected and screened. According to the evaluation criteria and tools of Cochrane,the heterogeneity among the studies was analyzed,and the Meta-analysis was carried out by Rev Man 5.3 software. A total of 16 studies were included in this study,with a total sample size of 1 788 cases,including 901 cases in the experimental group and 887 cases in the control group. Among them,11 intervention measures were Yupingfeng Granules combined with Western medicine routine vs routine Western medicine,and 5 research interventions were Yupingfeng Granules combined with pidomod vs pidomod based on routine therapy. Meta-analysis showed that the total effective rate of Yupingfeng Granules combined with Western medicine in the treatment of recurrent respiratory tract infection was better than that of Western medicine( RR= 1. 27,95%CI[1. 21,1. 34],P<0. 000 01). Based on the routine therapy,the total effective rate of Yupingfeng Granules combined with pidomod in the treatment of recurrent respiratory tract infection was better than that of pidomod( RR = 1. 23,95% CI[1. 13,1. 35],P<0. 000 01). A total of 21 cases of adverse events were reported in this study,including 6 cases in the experimental groups combined with traditional Yupingfeng Granules therapy and conventional Western medicine therapy and 15 in the control group. The adverse events were mainly gastrointestinal reactions,such as fatigue,nausea and diarrhea,which can tolerate or disappear with no impact on treatment. The incidence of adverse events in the two groups was not statistically significant( RR = 0. 44,95% CI[0. 19,1. 03],P = 0. 06). The results showed no significant difference in the incidence of adverse events between the two groups during treatment. Based on the routine therapy,no adverse event was reported in Yupingfeng Granules pidomod vs pidomod,indicating that both groups were safe. The analysis showed that Yupingfeng Granules combined with routine Western medicine therapy or combined with pidomod could increase the level of immunoglobulin and T lymphocytes in children. Based on the existing data and methods,Yupingfeng Granules combined with routine Western medicine therapy for recurrent respiratory tract infection can improve the total effective rate,immune function and body immunity,with no serious adverse reaction. However,because of the low quality of the literatures included in this study,it is still necessary to adopt well-designed large-sample clinical trials in conformity to international standards to improve the quality of evidence.

Regulatory Effect of Guben Fangxiao Decoction on Expression of TLR4 and TLR7 in Mice with Asthma Remission / 中国实验方剂学杂志

Objective:To observe the regulatory effect of Guben Fangxiao decoction on Toll-like receptor (TLR)4, TLR7, nuclear factor-kappa B p65 (NF-κB p65) and NF-κB inhibitor protein alpha (IκBα) in mice with asthma remission, in order to explore the mechanism of Guben Fangxiao decoction in treating asthma remission. Method:Respiratory syncytial virus (RSV) combined with chicken ovalbumin (OVA) was used to build asthma remission model in 3-week-old BALB/c mice. Sixty mice were divided into blank group, model group, dexamethasone group (0.001 g·kg-1), low,medium and high-dose Guben Fangxiao decoction group (6.5,13,26 g·kg-1), respectively. Intervention was given once a day for 28 days. After administration, the mice were put to death. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissue and score the pulmonary inflammation. Western blot was used to detect the expressions of TLR4, TLR7, IκBα and nuclear NF-κB p65 in lung tissue of mice. Immunofluorescence was used to observe the expression of NF-κB p65 in cellnucleuses. Result:Compared with blank group, the lung tissue of model group showed obvious inflammatory cell infiltration (PκB p65 increased significantly (PκBα proteins increased significantly (PPκB p65 (PκB p65 (PκBα proteins (PConclusion:Guben Fangxiao decoction can alleviate airway inflammation, and its therapeutic effect may be achieved by regulating TLR4, TLR7, NF-κB p65 and IκBα.

Intervention of Modified Yupingfeng San Combined with Aspirin on D-dimer， PagT Levels of RSA with Seronegative APS / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of aspirin combined with modified Yupingfeng San on patients with recurrent spontaneous abortion and Seronegative antiphospholipid syndrome. Method:From December 2016 to March 2018, 109 cases of recurrent spontaneous abortion patients with seronegative antiphospholipid syndrome in Affiliated Hospital of Nanjing University of Chinese Medicine were selected as the study objects. According to the random number table, they were divided into three groups:blank group, control group and study group. The blank group was treated with folic acid tablets, 1 pill/time, qd, as placebo, the control group was treated with folic acid tablets, 1 pill/time, qd, Danshen pills, 4 pills/time, tid, and aspirin, 1 pill/time, tid, and the study group was treated with folic acid tablets, 1 pill/time, qd, aspirin, 1 pill/time, tid, and modified Yupingfeng San, 150 mL/time, bid. These patients were all asked for pregnancy preparation for 1 year and treated with aspirin, 1 pill/time, tid, and progesterone pills, 1 pill/time, tid, since the beginning of pregnancy until 13 weeks or abortion. The pregnancy rate, miscarriage rate, coagulation index before and after treatment, traditional Chinese medicine (TCM) symptom score as well as months of abortion were all observed and compared. Result:The coagulation index and TCM symptom score of the study group were significantly lower than those before treatment (PPPPPConclusion:The data proved that the combination of anti-coagulation medicine with either single herb or herbal compound can improve the curative effect of the patients, and traditional Chinese herbal compound can regulate the immune system, reduce the thrombosis and improve the symptoms of TCM, which made the curative efficacy more prominent. However, more samples were needed for further research. Because the time of fetal protection may not change the outcome of pregnancy, the therapy should be maintained at least until the end of the early pregnancy.

Comprehensive Optimization of Rehabilitation Measures of Traditional Chinese Medicine to Elderly Patients with COPD at Stable Period / 中国实验方剂学杂志

Objective:To discuss the clinical efficacy of comprehensive optimization of rehabilitation measures of traditional Chinese medicine(TCM) on elderly patients with COPD at stable period, in order to study its effect on immune function and inflammatory factors. Method:One hundred and forty-four patients were randomly divided into control group and observation group by random number table. Patients in control group got ipratropium bromide, 40-80 μg/time, 2-4 times/days, salmeterol roticasone powder, 1 suction/time, 2 times/days, as well as comprehensive western medical treatment measures of healthcare education, oxygen therapy and respiratory muscle exercise. In addition to the basic therapy of ipratropium bromide and salmeterol roticasone powder, patients in observation group was added with syndrome differentiation therapy of TCM, foot bath with TCM and respiratory function exercise with TCM. The course of treatment was 6 months, and 6-month follow-up were recorded. Comprehensive assessment of COPD was recorded. Before and after treatment, respiratory questionnaire (mMRC), self-assessment test for patients with chronic obstructive pulmonary disease (CAT), forced expiratory volume in the first second (FEV1), FEV1%, St George's respiratory questionnaire (SGRQ), 6-minute walking distance (6 MWD) were scored. And levels of CD3+, CD4+, CD8+, CD4+/CD8+, interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha) and hypersensitive C-reactive protein (hs-CRP) were detected. Result:Comprehensive assessment of COPD in the two groups were better than those before treatment (PZ=2.066, Pα were lower than those in control group (P+ was lower than that in control group (P1 and FEV1% were higher than those in control group (P+, CD4+, CD4+/CD8+ and 6-minute walking distance were more than those in control group (PConclusion:Comprehensive optimization of rehabilitation measures in TCM can inhibit inflammatory factors, improve immune function, alleviate clinical symptoms, improve lung function, improve exercise endurance and quality of life of patients, and promote the lung rehabilitation.

Curative effect of Yupingfeng combined with nutritional support therapy on children with hand-foot-mouth disease / 中草药

Objective To analyze the clinical efficacy of Yupingfeng Granules combined with nutrition support for the treatment of children with hand-foot-mouth disease (HFMD) reinfection. Methods A total of 100 children with reinfection of HFMD were randomly divided into treatment group and control group. The treatment group was given Yupingfeng Granule or oral liquid combined with individualized nutritional support on the basis of routine treatment, while the control group was given routine treatment only. Results There was no significant difference in total effective rate between the treatment group and the control group in 50 children with reinfection of HFMD (P > 0.05). But fever subsided time, rash subsided time, and recovery in treatment group were shorter than those in the control group, the difference was statistically significant (P < 0.05). Conclusion Yupingfeng combined nutrition support has good clinical effects on children with HFMD reinfection.

Effect of Yupingfeng granule on Th17/Treg balance in children with milk protein allergy / 中国医师杂志

Objective To observe Thl7 cells,Treg cells and their related factors in peripheral blood of children with milk protein allergy and explore the influence of Yupingfeng granule on Th17 / Treg imbalance and its clinical effect.Methods 40 children with milk protein allergy were divided into two groups randomly:the conventional treatment group (n =20),Yupingfeng granule group (n =20).The conventional treatment group received conventional treatment for 2 months.On the basis of routine treatment,Yupingfeng granule group was additionally treated with Yuping feng granule.The serum Th17,Treg cell counts,interleukin (IL)-17 and transforming growth factor-β1 (TGF-β1) levels were detected,and the eczema area and severity index (EASI) score of rash in children was recorded.Results The levels of Th17 cells and IL-17 in allergy children were obviously increased compared with those of the normal children,while the Treg cells,and the TGF-β1 level were lower than those of the normal children (P ＜ 0.05).After treatment,the Thl7 cells,the IL-17 levels and ESAI scores of the conventional treatment group and the Yupingfeng granule group were lowered,while the Treg cells and the TGF-β1 levels were increased (P ＜ 0.05).Compared with the conventional treatment group,these indexes increased and decreased more significantly in the Yupingfeng granule group (P ＜ 0.05).Conclusions Milk allergy children have obvious imbalance of Thl7/Treg;the Yupingfeng granule can adjust this imbalance and alleviate the allergic symptoms of milk protein.

Meta-analysis of Efficacy and Safety of Yupingfeng Powder Combined with Second-generation Antihistamines Versus Second-generation Antihistamines for Chronic Urticaria / 中国药房

OBJECTIVE:To evaluate the efficacy and safety of Yupingfeng powder combined with second- generation antihistamines versus second-generation antihistamines for chronic urticaria(CU)systematically,and to provide evidence-based reference for clinical treatment for CU. METHODS:Retrieved from PubMed,Embase,The Cochrane Library,CJFD,VIP and CBM,RCT about therapeutic efficacy(total response rate,cure rate,recurrence rate)and safety(the incidence of ADR)of Yupingfeng combined with second-generation antihistamines(trial group)versus second-generation antihistamines(control group) in the treatment of CU were collected. The data extraction was performed for included clinical studies,and Meta-analysis was performed by using Rev Man 5.3 statistical software after quality evaluation with Cochrane Handbook 5.1.0 evaluation criteria. RESULTS:A total of 34 RCTs were enrolled,involved 3 405 patients in total. Results of Meta-analysis showed that the total response rate [OR=4.02,95%CI(3.03,5.34),P<0.001],cure rate [OR=2.25,95%CI(1.95,2.60),P<0.001] and recurrence rate [OR=0.33,95%CI(0.26,0.42),P<0.001] of trial group were significantly better than those of control group,with statistical significance. There was no statistical significance in the incidence of ADR between 2 groups [OR=0.98,95%CI(0.71,1.37),P=0.92]. CONCLUSIONS:For CU therapy,Yupingfeng powder combined with second-generation antihistamines is better than second-generation antihistamines alone in improving total response rate and cure rate,reducing recurrence rate,both have similar safety.