Amygdalin Improves Allergic Asthma via the Thymic Stromal Lymphopoietin-dendritic Cell-OX40 Ligand Axis in a Mouse Model.

Asthma, characterized by persistent inflammation and increased sensitivity of the airway, is the most common chronic condition among children. Novel, safe, and reliable treatment strategies are the focus of current research on pediatric asthma. Amygdalin, mainly present in bitter almonds, has anti-inflammatory and immunoregulatory potential, but its effect on asthma remains uninvestigated. Here, the impact of amygdalin on the thymic stromal lymphopoietin (TSLP)-dendritic cell (DC)-OX40L axis was investigated. A BALB/c mouse model for allergic asthma was established using the ovalbumin-sensitization method. Amygdalin treatment was administered between days 21 and 27 of the protocol. Cell numbers and hematoxylin and eosin (H&E) staining in bronchoalveolar lavage fluid (BALF) were used to observe the impact of amygdalin on airway inflammation. TSLP, IL-4, IL-5, IL-13, and IFN-γ concentrations were determined via Enzyme-linked immunosorbent assay (ELISA). TSLP, GATA-3, and T-bet proteins were measured using western blotting. Cell-surface receptor expression on DCs (MHC II, CD80, and CD86) was assessed via flow cytometry. OX40L mRNA and protein levels were detected using western blotting and qRT-PCR, respectively. Amygdalin treatment attenuated airway inflammation decreased BALF TSLP levels, inhibited DC maturation, restrained TSLP-induced DC surface marker expression (MHCII, CD80, and CD86), and further decreased OX40L levels in activated DCs. This occurred together with decreased Th2 cytokine levels (IL-4, IL-5, and IL-13) and GATA3 expression, whereas Th1 cytokine (IFN-γ) levels and T-bet expression increased. Amygdalin thus regulates the Th1/Th2 balance through the TSLP-DC-OX40L axis to participate in inflammation development in the airways, providing a basis for potential allergic asthma treatments.

The "time-response" effect of Wenyang Pingchuan Formula on miR-19a in asthmatic mice experiment.

BACKGROUND: Wenyang Pingchuan Formula (WPCF) is an empirical formula for the treatment of acute childhood asthma. However, the "time-effect" relationship of this prescription is not clear. This paper explores the relationship between Janus activated kinase signal transducer and activator of transcriptions (JAK/STAT), nuclear factor-κB (NF-κB), and microRNA (miR-19a), and also preliminarily determines the best time-effect relationship of WPCF in reducing the airway inflammation in asthmatic mice. METHOD: 80 BALB/c mice were randomly divided into four groups: control (CON) group, model (MDL) group, dexamethasone (DEX) group, and WPCF group. MDL group was established through intraperitoneal injection of 10% ovalbumin (OVA) and Al(OH)3 solution and the inhalation of aerosolized 5% OVA solution. Enzyme-linked immunosorbent assay (ELISA), real-time PCR and Western blot were conducted to determine the levels of interleukin (IL)-4, IL-13, interferon-γ (IFN-γ) in bronchoalveolar lavage fluid (BALF), contents of miR-19a mRNA and STAT6, phosphorylated signal transducers and activators of transcription 6 (p-STAT6), p65, phosphorylated p65 (p-p65), suppressors of cytokine signaling 1 (SOCS1), and tumor necrosis factor α-induced protein-3 (Tnfaip3) proteins after 7 and 28 days of intervention respectively. RESULTS: Significant down-regulation of IL-4 and IL-13 expression (P<0.05) and up-regulation of IFN-γ expression (P<0.05) in BALF have been observed for WPCF group compared with the MDL group. The significant down-regulation of miR-19a mRNA and STAT6, p-STAT6, p65, p-p65 proteins (P<0.05) and up-regulation of SOCS1 and Tnfaip3 proteins (P<0.05) in BALF was also observed for WPCF group compared to the MDL group. During the experiment, the weight of the mice in DEX group significantly decreased (P<0.05) compared with the other groups. CONCLUSIONS: WPCF could restore Th1/Th2 balance. The longer the intervention time, the more effective the treatment. The down-regulation of miR-19a mRNA by activating JAK/STAT and NF-κB signal pathways may be a possible mechanism by which WPCF alleviates airway inflammation.

Pingchuan formula improves allergic asthma in mice through inhibiting nuclear factor-kappa B / mitogen-activated protein kinase signaling pathway.

OBJECTIVE: To examine the role and decipher the mechanism of Pingchuan formula (PCF) in treating allergic asthma. METHODS: The mice were treated with saline, dexamethasone (DXM) and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance (RI), pulmonary dynamic compliance (Cdyn) and maximum voluntary ventilation (MVV) were measured. In addition, cellular changes in bronchoalveolar lavage fluid (BALF) and pathological changes in lung biopsy as well as the expression level of -smooth muscle actin (α-SMA), transforming growth factor-beta1 (TGF-α1) in BALF and interleukin-5 (IL-5), interleukin-13 (IL-13), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), nuclear factor-kappa B-p65 (NF-κBp65), inhibitor-α of nuclear transcription factor κB (IκBα), p38 mitogen-activated protein kinase (p38MAPK), c-jun n-terminal kinase (JNK) and its phosphorylated proteins in lung tissue were also examined and compared among different groups. RESULTS: Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group. Both DXM and PCF effectively decreased the number of eosinophils, lymphocytes, and neutrophils in BAL as well as the secretion of α-SMA and TGF-α1, IL-5, IL-13, while increased the expression of TNF-α and IFN-γ. Furthermore, our study indicated that the NF-κBp65, IκBα, p38MAPK and JNK pathways were inhibited under the treatment of PCF. CONCLUSION: Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway. This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of thediseasepathogenesis.

Pingchuan formula attenuates airway mucus hypersecretion via regulation of the PNEC-GABA-IL13-Muc5ac axis in asthmatic mice.

BACKGROUND/AIMS: Asthma is a common chronic respiratory disease. It has been reported that Pingchuan formula (PCF) can control asthma attacks by reducing airway inflammation, muscle spasm and mucus secretion. However, PCF's mechanism for reducing airway mucus hypersecretion remains unclear. This study aimed to investigate the effect of PCF on airway mucus secretion in asthmatic mice and to explore changes in the PNEC-GABA-IL13-Muc5ac axis. METHODS: Male Babl/c mice were used to establish the asthma model via sensitisation with OVA. Mice were randomly divided into Normal, OVA, DEX, and PCF groups. After treatment, lung histopathology was observed with H&E and PAS staining. BALF levels of IL-5 and IL-13 were detected using ELISA. The levels of mRNA and protein expression for GAD1, GABAARß1, GABAARα1 and Muc5ac in the lung tissue were measured by RT-PCR and Western blot assays. PNECs were observed with AgNOR staining. RESULTS: PCF treatment effectively reduced goblet cell (P < 0.01) and PNEC (P < 0.05) proliferation, lung tissue inflammation and airway mucus hypersecretion. In addition, PCF also markedly downregulated mRNA and protein expression of GAD1, GABAARß1, GABAARα1 and Muc5ac (P < 0.05, compared with OVA), thus inhibiting the GABA-IL-13 pathway in the lung tissue of asthmatic mice. CONCLUSION: These findings suggest that PCF controls asthma attacks by reducing airway inflammation and mucus hypersecretion via the PNEC-GABA-IL13-Muc5ac axis.

Study of item text in the Chinese Symptom Checklist-90.

ABSTRACT: Symptom Checklist-90 (SCL-90) is the popular and widely used instrument, whether in mental health screening, psychological consultation, or the diagnosis and estimate of mental illness. In 1984, it was translated from theEnglish version into Chinese and then SCL-90 has been widely used in China. It is a pity that the item text of Chinese version has not been revised since the birth of it until today. We analyzed the Chinese version of the 90-item text from 3 new perspectives: translation, semantic, and cross-cultural, and thought that 18 items should be revised. This study' results have taken one step forward on the basis of previous studies, which will play an important role in improving the quality of Chinese version SCL-90 and improving the mental health level of Chinese people.

Meta-analysis of the Unintentional Death of Children Aged 0 to 5 Years in China Between 1995 and 2015.

OBJECTIVES: The aim of this study was to identify the primary cause of and a means by which to prevent the unintentional injury and death of children aged 0 to 5 years in China. METHODS: A systematic review of literature published in this area between 1995 and 2015, accessible via Chongqing VIP, Wanfang, CNKI, and PubMed databases, in addition to other electronic databases and related magazines and professional information, was conducted. The data were retrieved using Cochrane Systematic review methods. All information was judged using the Jadad method. A meta-analysis was carried out on homogeneous studies. RevMan 5.3 software was used to analyze the gathered information. RESULTS: A total of 125 related pieces of literature were retrieved. Of the 14 that met the inclusion criteria, 6 were of moderate quality and 8 of general quality. The meta-analysis showed that in a comparison of unintentional injury and death of male and female children, the odds ratio was [1.73 to 1.95] for a 95% confidence interval (P < 0.00001). In a comparison of urban and rural areas, the odds ratio was [0.02, 0.02] for a 95% confidence interval (P < 0.00001). CONCLUSIONS: Based on accessible literature, the incidence of unintentional injury and death of boys is higher than that in girls and is also higher in rural areas than in urban areas of China. Considering the significant difference in the latter comparison, developing and implementing methods to reduce the unintentional injury and death of children in rural areas should receive greater attention.

Therapeutic effects of liver soothing pingchuan formula decoction on experimental asthma in BALB/c mice via regulation of nerve growth factor-tyrosine kinase A pathway.

The present study was designed to investigate the effects of liver soothing pingchuan formula decoction (LSPF) on experimental asthma in BALB/c mice and explore its potential molecular mechanisms. An animal model of asthma was established in BALB/c mice through sensitization and activation with intraperitoneal injection of 10% ovalbumin (OVA)/Al(OH)3 solution in addition to inhalation of a 5% OVA solution. LSPF (300 and 600 mg/kg/day) was initially administered orally prior to activation. Following this, bronchoalveolar lavage fluid (BALF) and lung tissues were collected for histopathalogical examination. Levels of inflammatory cells and cytokines were determined in the BALF, and levels of nerve growth factor (NGF) and tyrosine kinase A (TrkA) in the lung tissues were determined. The results of the present study indicated that increased inflammatory reactions were observed following OVA sensitization (P<0.05), and the expression levels of NGF (P<0.05) and TrkA (P<0.05) were significantly increased, compared with normal mice. Notably, compared with the asthma model group, immunohistochemical results revealed that LSPF treatment suppressed OVA induced inflammatory reactions (P<0.05) and NGF (P<0.05) and TrkA expression levels (P<0.05). In addition, the NGF (P<0.05) and TrkA (P<0.05) were revealed to be downregulated with LSPF treatment from the results of the ELISA and western blotting assay. Overall, the results of the present study demonstrated that LSPF exhibits therapeutic effects on experimental asthma in mice, via downregulation of the NGF-TrkA pathway.

Effect of treatment with geraniol on ovalbumin-induced allergic asthma in mice.

BACKGROUND: Asthma, a complex highly prevalent airway disease, is a major public health problem for which current treatment options are inadequate. OBJECTIVE: To evaluate the antiasthma activity of geraniol and investigate its underlying molecular mechanisms. METHODS: In a standard experimental asthma model, Balb/c mice were sensitized with ovalbumin, treated with geraniol (100 or 200 mg/kg) or a vehicle control, during ovalbumin challenge. RESULTS: Treatment of ovalbumin-sensitized/challenged mice with geraniol significantly decreased airway hyperresponsiveness to inhaled methacholine. Geraniol treatment reduced eotaxin levels in bronchoalveolar lavage fluid and attenuated infiltration of eosinophils induced by ovalbumin. Geraniol treatment reduced TH2 cytokines (including interleukins 4, 5, and 13), increased TH1 cytokine interferon γ in bronchoalveolar lavage fluid, and reduced ovalbumin-specific IgE in serum. In addition, treatment of ovalbumin-sensitized/challenged mice with geraniol enhanced T-bet (TH1 response) messenger RNA expression and reduced GATA-3 (TH2 response) messenger RNA expression in lungs. Furthermore, treatment of ovalbumin -sensitized/challenged mice with geraniol further enhanced Nrf2 protein expression and activated Nrf2-directed antioxidant pathways, such as glutamate-cysteine ligase, superoxide dismutase, and glutathione S-transferase, and enhanced formation of reduced glutathione and reduced formation of malondialdehyde in lungs. CONCLUSION: Geraniol attenuated important features of allergic asthma in mice, possibly through the modulation of TH1/TH2 balance and activation the of Nrf2/antioxidant response element pathway.

Ameliorative effects of Qingfei Tongluo formula on experimental mycoplasmal pneumonia in mice.

Mycoplasma pneumoniae pneumonia (MPP) is a common disease in children. Qingfei Tongluo formula (QTF) has been used for the treatment of MPP clinically, but the chemical constituents and mechanism involved remain unclear. This study aimed to analyze the main chemical constituents and to explore the possible mechanism of action associated with QTF treatment of MPP. Liquid chromatography-mass spectrometry was employed to identify the compounds contained in the QTF extract. A BALB/c mouse model of MP infection was established. After treatment with QTF (0.85 and 1.70 g/kg) for 3 days, hematoxylin and eosin staining was performed in lung tissues for histological examination. Inflammatory cytokines were detected by ELISA. Western blot analysis was used for detecting phosphorylated proteins involved in MAPK and nuclear factor (NF)-κB signaling pathways. In the mouse model, a large amount of pulmonary interstitial infiltration of lymphocytes and plasmacytes were seen as well as bronchus and vasodilation congestion. Following QTF treatment, inflammation was alleviated significantly compared with the model group. Inflammatory cytokines [interleukin (IL)-6, transforming growth factor-ß1, IL-8, IL-1ß and tumor necrosis factor-α] in bronchoalveolar lavage fluid were decreased dramatically. In addition, we found that QTF inhibited activation of phosphorylation of JNK, ERK and NF-κB. In conclusion, QTF alleviates MPP inflammation possibly via inhibitory activation of MAPK/NF-κB pathways, which can act as a new agent for MPP treatment.

[Effects of Chinese herbal medicine Pingchuan Formula on airway inflammation, interferon-γ and interleukin-4 in mice with asthma].

OBJECTIVE: To observe the changes of interferon-γ (IFN-γ) and interleukin-4 (IL-4), factors related to T helper 1/T helper 2 (Th1/Th2), in airway inflammation in an asthmatic model of mice and the effects of Pingchuan Formula (PCF), a compound traditional Chinese herbal medicine. METHODS: Eighty BALB/c mice were randomly divided into normal group, model group, dexamethasone group and PCF group, with 20 mice in each group. For inducing asthma, the BALB/c mice were sensitized by intraperitoneal injection of mixed ovalbumin, and then activated by inhaling ovalbumin. After 2 weeks of sensitizing, asthmatic mice were intragastrically administered with the drugs once a day for four weeks. Bronchoalveolar lavage fluid (BALF) and lung tissue were collected respectively after 7-day activation or 2 weeks after activation. The general histological changes were observed by hematoxylin and eosin staining and collagen staining, meanwhile the levels of IFN-γ and IL-4 in BALF were detected by enzyme-linked immunosorbent assay. RESULTS: After 7 d of activation, compared with the normal group, significant decrease in IFN-γ and increase in IL-4 were detected in the model group; compared with the model group, IL-4 level of the two treated groups was decreased and the IFN-γ/IL-4 ratio was increased significantly; no significant difference was found between the two treated groups. Two weeks after activation, compared with the normal group, IFN-γ was decreased, IL-4 was increased, and the IFN-γ/IL-4 ratio was decreased in the model group; compared with the model group, IL-4 level in the dexamethasone group was significantly decreased and IFN-γ/IL-4 ratio was significantly elevated, while the level of IL-4 in the PCF group was significantly decreased, the level of IFN-γ was increased simultaneously, and the IFN-γ/IL-4 ratio was significantly elevated. There was no significant difference between the two treated groups in IFN-γ and IL-4 levels, nor in the IFN-γ/IL-4 ratio. The area of collagen fiber in airway wall epithelium in the model group was smaller than that in the normal group, and those in the treated groups were lower than that in the model group; there was no significant difference between the two treated groups. CONCLUSION: PCF has regulating function in correcting the imbalance of Th1/Th2 so as to improve airway inflammation. The therapeutic effects of PCF are basically similar to dexamethasone.

[Effects of Chinese herbal medicine Bushen Gubiao Recipe on toll-like receptor 4 and CD4(+)CD25(+)foxp3(+) regulatory T cells in mice with recurrent respiratory tract infections].

OBJECTIVE: To evaluate the effects of Bushen Gubiao Recipe (BGR), a compound traditional Chinese herbal medicine, on toll-like receptor 4 (TLR4) signaling pathway and CD4(+)CD25(+)foxp3(+)regulatory T cells (CD4(+)CD25(+)foxp3(+)Tregs) in mice with recurrent respiratory tract infections (RRTIs). METHODS: A mouse model of kidney-yang deficiency which simulated physical characteristics of RRTIs was established by intraperitoneal injection of hydrocortisone for 14 d. The model mice were divided into 4 groups, model group, high-dose BGR group, low-dose BGR group, and nucleic acid and casein oral solution group. They were administered respectively with distilled water, high-dose BGR (50 g/kg body weight), low-dose BGR (25 g/kg body weight) and nucleic acid and casein oral solution. Besides, a normal control group was set up and gastrogavage with distilled water. The effect of intervention was evaluated 4 weeks later by estimating the changes in behaviors of mice. Expressions of TLR4 and nuclear factor-kappa B (NF-κB) p65 in lung tissue were detected by immunohistochemical SABC method, the expression of TLR4 mRNA in lung tissue was detected by fluorescence quantitative-polymerase chain reaction (FQ-PCR), and the level of blood CD4(+)CD25(+)foxp3(+) Tregs was determined by flow cytometry. RESULTS: A kidney-yang deficiency mouse model with RRTIs was successfully established by intraperitoneal injection of hydrocortisone. BGR could improve the abnormal behavioral condition of the mice and enhance the expressions of TLR4 and NF-κB p65 in the lung tissue. Expression of TLR4 mRNA in high-dose BGR group was higher than that in model group (P<0.05), while the difference was not statistically significant between high-dose BGR group and low-dose BGR group (P>0.05). Levels of CD4(+)CD25(+)foxp3(+)Tregs in high-dose BGR group and nucleic acid and casein oral solution group were lower than that in model group (P<0.05), while the difference was not statistically significant between high-dose BGR group and low-dose BGR group (P>0.05). CONCLUSION: BGR can improve the behavior of the kidney-yang deficiency mice, and improve the innate immune function by up-regulating TLR/NF-κB signaling pathway. BGR can adjust the immune imbalance of T-helper cell (Th) 1/Th2 through reducing the activity of CD4(+)CD25(+)foxp3(+)Tregs and enhancing the immune response of Th1 type.

[Effects of Pingchuan Recipe on airway remodeling in mice with bronchial asthma].

OBJECTIVE: To study the effects of Pingchuan Recipe, a compound traditional Chinese herbal medicine for treating bronchial asthma, on macrophage inflammatory protein-1alpha (MIP-1alpha) and immunoglobulin E (IgE) contents and CD86 expression in a mouse model of bronchial asthma, and to investigate the mechanism of Pingchuan Recipe in regulating airway remodeling in mice with bronchial asthma. METHODS: Fifty BALB/c mice were randomly divided into normal control group, untreated group, dexamethasone group, low-dose Pingchuan Recipe group and normal-dose Pingchuan Recipe group. Bronchial asthma in mice was induced by intra-abdominal injection of mixed ovalbumin and stimulation by inhaling 5% ovalbumin. Mice in the normal control group and the untreated group did not receive treatment, but mice in the other groups were administered intragastrically with 50 g/ (kg * d) Pingchuan Recipe, 25 g/ (kg * d) Pingchuan Recipe and 0.75 mg/ (kg * d) dexamethasone, respectively. After consecutive treatment for 8 weeks, the MIP-1alpha contents in peripheral blood and bronchoalveolar lavage fluid (BALF) were measured by an avidin biotin complex-enzyme-linked immunosorbent assay (ABC-ELISA) system, and the IgE contents in serum and BALF were measured by ELISA method. Direct immunofluorescence-flow cytometry was used to detect the percentages of CD86(+) and CD3(-) CD86(+) cells in peripheral blood and BALF. General histological changes of the lung tissue were observed by HE staining and collagen fiber staining 12 weeks after activation. RESULTS: Compared with the normal control group, the contents of MIP-1alpha, the percentages of CD86(+) cells and CD3(-) CD86(+) cells in peripheral blood and BALF, and the contents of IgE in serum and BALF were raised significantly (P < 0.01). The contents of MIP-1alpha, the percentages of CD86(+) cells and CD3(-) CD86(+) cells in peripheral blood and BALF, and the contents of IgE in serum and BALF in the treated groups were lower than those in the untreated group (P < 0.01). The contents of MIP-1alpha in peripheral blood and BALF and the contents of IgE in serum and BALF had no significant differences among the treated groups (P > 0.05). The percentage of CD3(-) CD86(+) cells in peripheral blood in low-dose Pingchuan Recipe group was lower than that in the dexamethasone group. Compared with the untreated group, the collagen deposition in the wall of bronchiole in the treated groups was decreased. CONCLUSION: Pingchuan Recipe can decrease the contents of MIP-1alpha, IgE and the expressions of CD86 in peripheral blood and BALF, which may be one of its mechanisms in improving chronic airway inflammation and inhibiting airway remodeling.

Multi-centered clinical study on effects of nano-amomi paste in treating children's anorexia.

OBJECTIVE: To assess the effect and safety of nano-Amoni Paste (nmAP) in the treatment of children's anorexia (AR). METHODS: One hundred and eighty patients of AR were assigned according to the randomized, double-blinded, double-simulated and parallel controlled principle to three groups, the treated group (TG), the positive control group (PCG) and the negative control group (NCG), 60 in each group. The patients in TG were treated by sticking 1.5 ml of nmAP on the acupoint of Shenque (Ren 8) once a day and orally taking placebo liquid 10 ml twice a day; those in PCG and NCG treated with sticking paste of placebo on Ren 8, and oral taking of Shanmai Jianpi Oral Liquid and placebo liquid respectively, 10 ml each time twice per day. The course of treatment for all was 10 days, all patients were treated for 2 courses. RESULTS: The total effective rate and the effective rate on cardinal symptom in TG was 85.0% and 95.0% respectively, that in PCG 86.2% and 96.55% and in NCG 45.5% and 65.45%, respectively, showing significant difference between groups (P<0.05). Comparison of the clinical manifestation before and after treatment showed significant improvement in volume of food intake, appetite, complexion and reduction of restlessness symptom (P<0.05) in all three groups, and there was no adverse reaction found in them. CONCLUSION: nmAP is an effective and safe remedy for treatment of Children's anorexia.

[Therapeutic effect of Xincang Decoction on chronic airway inflammation in children with bronchial asthma in remission stage].

OBJECTIVE: To observe the therapeutic effect of Xincang Decoction on chronic airway inflammation in children with asthma in clinical investigation. METHODS: Xincang Decoction was composed of Flos Magnoliae (Xinyi) and Fructus Xanthii (Cangoerzi), the traditional Chinese herbs for expelling wind. Sixty cases of children with bronchial asthma in remission stage were randomly divided into two groups. Thirty cases in the treatment group were treated with Xincang Decoction and the others in the control group were treated with ketotifen fumarate. The therapeutic effects of the two groups were compared, and the peripheral eosinophil (EOS) count, the levels of immunoglobulin E (IgE), interleukin 4 (IL-4) and interleukin 5 (IL-5), and the pulmonary functions were observed before and three months after the treatment. RESULTS: After three months treatment, the results showed that the total response rates of the treatment and the control group were 83.3% and 80.0%, respectively, without marked difference (P>0.05). The levels of EOS and IL-5 were obviously decreased after the treatment, and the levels of EOS and IL-5 of the patients in the treatment group were lower than those in the control group (P<0.05). Meanwhile the forced expiratory volume in one second (FEV(1)) was improved after the treatment, and the FEV(1) of the patients in the treatment group was higher than that of the patients in the control group (P<0.05). CONCLUSION: Xincang Decoction can decrease the levels of EOS and IL-5 and improve the pulmonary function in treating chronic airway inflammation in children with bronchial asthma in remission stage.