Investigating the effects of Laggera pterodonta on H3N2-Induced inflammatory and immune responses through network pharmacology, molecular docking, and experimental validation in a mice model.

For centuries, Laggera pterodonta (LP), a Chinese herbal medicine, has been widely employed for treating respiratory infectious diseases; however, the mechanism underlying LP's effectiveness against the influenza A/Aichi/2/1968 virus (H3N2) remains elusive. This study aims to shed light on the mechanism by which LP combats influenza in H3N2-infected mice. First, we conducted quasi-targeted metabolomics analysis using liquid chromatography-mass spectrometry to identify LP components. Subsequently, network pharmacology, molecular docking, and simulation were conducted to screen candidate targets associated with AKT and NF-κB. In addition, we conducted a series of experiments including qPCR, hematoxylin-eosin staining, flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assay to provide evidence that LP treatment in H3N2-infected mice can reduce pro-inflammatory cytokine levels (TNF-α, IL-6, IL-1ß, and MCP-1) while increasing T cells (CD3+, CD4+, and CD8+) and syndecan-1 and secretory IgA expression. This, in turn, aids in the prevention of excessive inflammation and the fortification of immunity, both of which are compromised by H3N2. Finally, we utilized a Western blot assay to confirm that LP indeed inhibits the AKT/NF-κB signaling cascade. Thus, the efficacy of LP serves as a cornerstone in establishing a theoretical foundation for influenza treatment.

Effect of Electroacupuncture at Zusanli (ST36) on Intestinal Microbiota in Rats With Chronic Atrophic Gastritis.

Background: Electroacupuncture is a common treatment for chronic atrophic gastritis (CAG) in China. We aimed to determine the effects of electroacupuncture at zusanli (ST36) on intestinal microbiota in CAG rats. Methods: In total, 42 SD rats were randomly divided into normal (NC, 10 rats) and model (MG, 32 rats) groups. Rats in the MG group were established as CAG disease models. After that, the rats in the MG group were randomly divided into CAG (10 rats), electroacupuncture (EA, 10 rats), and Vitacoenzyme (Vit, 10 rats) groups. Rats in the NC and CAG groups were subjected to a 30-min/d confinement for 4 weeks. Rats in the EA group were given electroacupuncture at zusanli for 30 min/d for 4 weeks. Rats in the Vit group were given Vitacoenzyme solution 10 ml/(kg d) for 4 weeks. Histopathological changes in the gastric mucosa were observed with hematoxylin and eosin staining, and the gene expression level of p53, Bcl-2, and c-myc was determined using the qPCR method. The 16S rDNA sequencing technique was used to determine structural changes and relative abundance expression of intestinal flora. Results: Compared with the NC group, gastric mucosal pathology in the CAG group revealed significant inflammatory infiltration, and the gastric mucosal lesions in the electroacupuncture group were improved remarkably; the expression of p53 and c-myc genes in the CAG group increased (p < 0.05), while the expression of Bcl-2 genes decreased (p < 0.05) in the EA group, that of p53 and c-myc genes decreased (p < 0.05), and that of Bcl-2 genes increased (p < 0.05). The abundance of bacteria such as Lactobacillus, Desulfobacterota, and Bacteroides pectinophilus group in the CAG group increased (p < 0.05), while that of bacteria such as Gastranaerophilales, Romboutsia, and Blautia decreased (p < 0.05). The relative abundance of Desulfobacterota and Helicobacter in the EA group decreased (p < 0.05), while that of probiotic bacteria such as Oscillospirales, Romboutsia, and Christensenellaceae increased (p < 0.05). Conclusion: Electroacupuncture at zusanli can promote the repair of pathological damage to the gastric mucosa in rats with CAG, and the mechanism might relate to the reduction in the relative abundance of harmful bacteria, increase in the relative abundance of intestinal probiotics, and regulation of the intestinal microbiota.

[Effect of electroacupuncture on small airway function in patients with stable chronic obstructive pulmonary disease].

OBJECTIVE: To observe the regulatory effect of electroacupuncture (EA) on small airway function and exercise tolerance in patients with stable chronic obstructive pulmonary disease (COPD). METHODS: A total of 62 patients with stable COPD were randomized into an observation group (31 cases, 1 case dropped off) and a control group (31 cases, 5 cases dropped off). On the base of routine medication and aerobic exercise, the patients of the two groups all received EA at Danzhong (CV 17), Rugen (ST 18), Guanyuan (CV 4), Zhongwan (CV 12), Tianshu (ST 25) and Yingchuang (ST 16). In the observation group, filiform needles were used and inserted perpendicularly, 3 mm in depth. In the control group, the placebo needling method was performed, in which the needle was not inserted through skin at each point. In both groups, electric stimulation with low-frequency electronic pulse instrument was exerted, with continuous wave, 2 Hz in frequency, lasting 30 min each time in the two groups. The treatment was given once every other day, 3 times a week, for 14 treatments totally. Before and after treatment, the following indexes were compared in patients between the two groups, i.e. the lung function indexes (forced expiratory volume in first second [FEV1], forced vital capacity [FVC], the ratio of FEV1 to FVC [FEV1/FVC], maximal voluntary ventilation [MVV], the percentage of maximal expiratory flow [MEF] at 25% of FVC exhaled [MEF25], MEF50 and MEF75 in predicted value), cardiopulmonary exercise test indexs (metabolic equivalent [METS], oxygen uptake per kg body weight [VO2/kg], minute ventilation [VE], the percentage of oxygen pulse [VO2/HR] in predictd value, maximal minute ventilation [VEmax], ventilatory equivalent for oxygen [VE/VO2], ventilatory equivalent for carbon dioxide [VE/VCO2]), 6-minute walk distance (6MWD), the total score of COPD assessment test (CAT), the modified British Medical Research Council (mMRC) score and COPD comprehensive grade. RESULTS: After treatment, FVC%, MVV%, MEF75%, MEF50%, VO2/kg%, METs%, VEmax, VO2/HR%, 6MW and the total CAT score were all improved as compared with those before treatment in the observation group (P<0.05, P<0.01). After treatment, MEF75% and the total CAT score were reduced as compared with those before treatment in the control group (P<0.05). After treatment, MVV%, MEF50%, VO2/kg%, METs%, VEmax and 6MWD in the observation group were all better than those in the control group (P<0.05, P<0.01). CONCLUSION: Electroacupuncture can improve the respiratory function and exercise tolerance in COPD patients through removing small airway obstruction and increasing ventilation.

La-based-adsorbents for efficient biological phosphorus treatment of wastewater: Synergistically strengthen of chemical and biological removal.

The present work demonstrated the invention of synergistically strengthen of chemical and biological removal of phosphorus (P) in biological wastewater treatment, which was achieved by exposure the bioreactors to different levels of La-based-adsorbents. We fabricated a high-performance La2O2CO3 micro-adsorbent (H-La2O2CO3) and added it into sequencing batch reactors. When activated sludge was exposed to 40 mg/L H-La2O2CO3 for 40 d, effluent total phosphorus (TP) concentration significantly decreased to approximately 0.18 mg/L, with the steady removal efficiency of 96.4%, which is superior to the biological phosphorus removal (BPR). The effect of H-La2O2CO3 dosages on P removal in biological wastewater treatment was also detailedly investigated. The H-La2O2CO3 adsorbent could not only capture P by chemical bonding itself, but also increased protein (PN) contents of extracellular polymeric substances (EPS) and changed the functional group of EPS to chemically adsorb P. Additionally, the results of 16s rDNA molecular analysis revealed that the species richness and microbial diversity varied with the different dosages of adsorbent. Sequence analyses showed that the appropriate concentration of H-La2O2CO3 addition increased the contents of several polyphosphate accumulating organisms (PAOs) at genus level in sludge.

Efficacy and safety of Chou-Ling-Dan granules in the treatment of seasonal influenza via combining Western and traditional Chinese medicine: protocol for a multicentre, randomised controlled clinical trial.

INTRODUCTION: Chou-Ling-Dan (CLD) (Laggerapterodonta) granules are an ethnic herbal medicine from Yunnan province of China. CLD granules have been used for the treatment of inflammatory conditions and feverish diseases in China, including seasonal influenza, but few evidence-based medicine (EBM) clinical studies have been conducted to assess its efficacy and safety in the treatment of influenza. Here, we performed an EBM clinical trial combining Western Chinese medicine and traditional Chinese medicine (TCM) evaluation systems to evaluate the efficacy and safety of CLD granules in the treatment of seasonal influenza. METHODS AND ANALYSIS: The study is designed as a multicentre, randomised, double-blinded, double-simulation, oseltamivir-controlled and placebo-controlled, parallel-design clinical trial. Eligible subjects (n=318) will be allocated after satisfying the criteria (Western medicine). Subjects will be randomised to receive CLD granules, oseltamivir, or a placebo for 5 days of treatment and with follow-up after treatment to record symptoms and signs and to collect pharyngeal/throat swabs and serum samples for detecting the virus and antibodies. At the same time, the syndrome differentiation criteria of TCM, such as tongue body, furred tongue and type of pulse, will be recorded as determined by doctors of both Western and Chinese medicine. Participants will be instructed to comply with the protocol and to keep a daily record of symptoms. The primary and secondary outcomes and safety indicators will be used to evaluate the efficacy and safety of CLD granules in the treatment of seasonal influenza based on both Western Chinese medicine and TCM evaluation systems. ETHICS AND DISSEMINATION: The CLD granules clinical trial will be conducted in accordance with the Declaration of Helsinki and Good Clinical Practice and has been approved by the Ethics Committee of the First Affiliated Hospital of Guangzhou Medical University. All participants must provide written informed consent. The results obtained will be disseminated at international medical conferences and in peer-reviewed publications. TRIAL REGISTRATION NUMBER: NCT02662426; Pre-results.

Amelioration of diabetic nephropathy in db/db mice treated with tibetan medicine formula Siwei Jianghuang Decoction Powder extract.

Siwei Jianghuang Decoction Powder (SWJH) documented originally in the Four Medical Tantras-Blue Glaze exhibited beneficial effects on diabetic nephropathy (DN) via combined synergistically action of multiple formula components including Curcumae longae Rhizoma, Berberidis dictyophyllae Cortex, Phyllanthi Fructus and Tribuli Fructus. This study investigated the effects of SWJH on DN in db/db mice and possible underlying mechanisms. The ten weeks old db/db mice treated with SWJH by intra-gastric administration once a day for 8 weeks. After 8 weeks, body weight, water and food intake of mice were recorded. The level of fasting blood glucose (FBG) was measured. Serum creatinine (Scr), blood urea nitrogen (BUN), urine microalbumin (UMAlb), serum uric acid (UA) and urinary albumin excretion (UAE) were detected. An enzyme-linked immunosorbent assay was performed to test serum vascular endothelial growth factor (VEGF) and transforming growth factor-ß1 (TGF-ß1). Real-time PCR and Western blot analysis were used to test mRNA and protein expression of hypoxia inducible factor-1α (HIF-1α), VEGF and TGF-ß1 in kidney tissue. SWJH treatment significantly reduced the levels of FBG, Scr, BUN, UMAlb, UA and UAE and retarded renal fibrosis. SWJH treatment further significantly reduced serum TGF-ß1 level and downregulated the expression of HIF-1α, VEGF and TGF-ß1 at both mRNA and protein levels. Principal component analysis and partial least squares regression and hierarchical cluster analysis demonstrated that SWJH treatment significantly ameliorated renal damage in DN mice. These consequences suggested that SWJH formulations were effective in the treatment of DN through regulating the HIF-1α, VEGF and TGF-ß1 overexpression.

Novel nor-monoterpenoid indole alkaloids inhibiting glioma stem cells from fruits of Alstonia scholaris.

BACKGROUND: Glioblastoma multiforme (GBM) is a highly aggressive and frequently recurrent malignant brain tumor, and to date, the clinically effective drugs against GBM remain scarce. Natural products play an important role in drug discovery, and might be the resource of antitumor agents for GSCs. Alstonia scholaris (L.) R. Br. is rich in monoterpenoid indole alkaloids (MIAs) and used extensively for treatment of tumor in the traditional medicine system of Asia. PURPOSE: To search for new MIAs with antitumor activity against glioma stem cells from clinical patients and explore their mechanism. METHODS: Compounds were obtained from the fruits of A. scholaris by chromatographic separation, including silica gel, Sephadex LH-20 and recrystallization. Their structures were elucidated by the use of UV, IR, NMR and MS spectra. The antitumor activity of the compounds against the glioma stem cells (GSC-3#, GSC-12#, GSC-18#) were investigated by phenotypic screening and MTS assays. Cell proliferation assay by BrdU immunofluorescence staining, and apoptosis assay by cleaved-caspase-3 immunofluorescence staining and real-time PCR assay. The soft-agar clonal formation assay was performed to determine the antitumor efficacy of the compounds in vitro. RESULTS: Two new nor-monoterpenoid indole alkaloids were isolated from the fruits of A. scholaris. They exhibited selective antitumor activity against glioma stem cells (GSC-3#, GSC-12#, GSC-18#) with IC50 values of 15-25 µg/ml. Furthermore, they inhibited GSCs proliferation, induced GSCs apoptosis by increasing the expression of TNF-α and cleavage of caspase-3, and significantly damaged colony forming capacity of GSCs. CONCLUSION: New nor-monoterpenoid indole alkaloids from the fruits of A. scholaris provide new type promising molecule for the selective killing of human glioma stem cells.

Effects of indole alkaloids from leaf of Alstonia scholaris on post-infectious cough in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaf of Alstonia scholaris (L.) R. Br. (Apocynaceae), a wide used ethic-medicine in many Asia and Africa counties, has also been recorded as the common traditional Chinese medicine for treatment of illnesses in respiratory system by Dai people. AIM OF THE STUDY: To provide experimental data of clinical adaption of total indole alkaloids (TA) from leaf of A. scholaris for treating post-infectious cough in phase II clinical trial. MATERIALS AND METHODS: To model post-infectious cough, all animals except control group were instilled intra-tracheal with lipopolysaccharide (LPS) (80 µg/50 µL/mouse), followed by subsequent exposure to cigarette smoke (CS) for 30 min per day for a total of 30 days. Mice were orally given TA at dose of 10, 25, 50 mg/kg, and four main alkaloids (Sch: scholaricine, Epi: 19-epischolaricine, Val: vallesamine, Pic: picrinine) once daily. Cellular infiltration was assessed in the broncho-alveolar lavage fluid (BALF). Expression of interleukin-6 (IL-6) and C-reactive protein (CRP) in the serum was determined, the superoxide dismutase (SOD) activity as well as malondialdehyde (MDA) content in the serum and homogenate were examined. Finally, histopathological examination in the lungs was assessed by H. E. staining. RESULTS: After administration of TA and four major alkaloids respectively, the symptoms of cough in mice were obviously attenuated. Total white blood cells (WBC) and neutrophils (NEU) amounts in BALF were reduced obviously and the pathological damage of lung was also attenuated. There was also significant reduction in IL-6, CRP, MDA and a marked improvement in SOD. CONCLUSIONS: The efficacy of indole alkaloids against post-infectious cough (PIC) was shown in the down-regulation of inflammatory cells, cytokines, and the balance of antioxidants. What's more, the pharmacological effects of TA were better than single indole alkaloid, which might be related to the synergic effect of four major alkaloids.

Inhibition of influenza virus via a sesquiterpene fraction isolated from Laggera pterodonta by targeting the NF-κB and p38 pathways.

BACKGROUND: Influenza virus poses serious threats to human health, especially human infection with avian influenza virus. Laggera pterodonta (DC.) Benth is a medicinal plant that is widely used in Traditional Chinese Medicine, especially in Yunnan province, and has been used to treat influenza, pharyngolaryngitis, and bronchitis. However, the compound(s) responsible for the activity and their mechanisms of action against the influenza virus remain to be elucidated. METHODS: L. pterodonta extract was fractionated, and the active fraction was identified as Fraction 14 (Fr 14). Fr 14 was further analysed and characterized by ultra-high-performance liquid chromatography hyphenated with quadrupole-time of flight mass spectrometry (UHPLC/Q-TOF-MS). The inhibitory effect against influenza virus was evaluated using a cytotoxicity assay. Then, cytokines and chemokines were detected by qRT-PCR and a bio-plex assay. Signalling pathways that inhibited the influenza virus were identified using a western blotting assay. RESULTS: The active fr 14 showed a wide spectrum of anti-influenza virus activity. The pharmacological mechanisms showed that Fr 14 acts on the early stage of virus replication (0-6 h). It inhibited the p38/MAPK pathway and then inhibited the NF-κB pathway and COX-2. Fr 14 also prevented the increased expression of cytokines and chemokines. CONCLUSION: This study demonstrated the preliminary mechanisms of fr 14 against the influenza virus. Fr 14 possessed antiviral and anti-inflammatory effects. L. pterodonta can be used to develop innovative antiviral drugs, and further studies will be performed to illustrate the detailed mechanisms.

[Effect of abdominal acupuncture therapy on the endocrine and metabolism in obesity-type polycystic ovarian syndrome patients].

OBJECTIVE: To observe the effect of abdominal acupuncture on the endocrine and metabolic level in obesity-type polycystic ovarian syndrome (PCOS) patients. METHODS: Eighty-six PCOS patients were randomly and equally divided into medication group and abdominal acupuncture group. Patients of medication group were treated with metformin (250 mg/time, t.i. d. in the 1st week, and 500 mg/time, t.i.d. thereafter) for 6 months,and those of abdominal acupuncture group were treated by abdominal acupuncture [Zhongwan (CV 12), Liangmen (ST 21), etc., once daily for 6 months]. Changes of the body height, body Mass index (BMI), waist-hip ratio (WHR), Ferriman-Galleey score (FGS), menstrual frequency (MF) and ovarian volume (OV) were determined. Serum luteotrophic hormone (LH), free testosterone (T), follicle-stimulating hormone (FSH) contents were detected with radioimmunoassay. Fasting blood glucose (FBG), fasting insulin (FIN), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), Homa insulin resistance index (IRI) were detected with chromatometry respectively. RESULTS: After the treatment, BMI, WHR, FGS and OV were reduced significantly in both medication and abdominal acupuncture groups (P < 0.05), while MF of the two groups increased evidently (P < 0.05), and the effects of abdominal acupuncture group were significantly superior to those of medication group in down-regulating BMI, WHR and upregulating MF (P < 0.05). Regarding the reproductive hormons, serum LH, LH/FSH and T levels in the two groups decreased significantly (P< 0.05), and the effect of abdominal acupuncture was superior to that of medication group in reducing serum T level (P < 0.05). Following the treatment, FBG, BG and FIN and INS contents 2 h after meal,and Homa IR in both medication and abdominal acupuncture groups all decreased considerably (P < 0.05), but without significant differences between them (P > 0.05). Regarding the blood lipid levels after the treatment, serum TC, TG, and LDL-C levels of the two groups decreased significantly (P < 0.05), while serum HDL-C level increased remarkably (P<0.05), without significant differences between the two groups in these indexes (P > 0.05). CONCLUSION: Abdominal acupuncture treatment can improve the endocrine and metabolic function of patients with obesity-type PCOS.