Relieving Sore Throat Formula Exerts a Therapeutic Effect on Pharyngitis through Immunoregulation and NF-κB Pathway.

Relieving Sore Throat Formula (RSTF) is a formula approved by the China Food and Drug Administration and has been used for the treatment of pharyngitis in clinic for many years. However, the potential pharmacological mechanism still remains unknown. We combined multiple methods including bioinformatics data digging, network pharmacology analysis, and pathway analysis to predict the potential target of RSTF. We verified our in silico prediction results with an in vivo/vitro antibacterial effect test, mouse phagocytic index test, proliferation, transformation, and migration of mouse spleen lymphocytes. Alteration of NF-κB pathway was determined by Western blotting, immunofluorescence, and PCR. The in vivo experiments demonstrated that the RSTF could significantly relieve the symptoms of pharyngitis. A rat saliva secretion test showed that RSTF can effectively relieve the xerostomia symptom. A phenol red excretion test showed that RSTF has an eliminating phlegm effect. A hot plate method and granuloma experiment proved that RSTF also have analgesic and anti-inflammatory effects. In silico prediction demonstrates that 70 active compounds of RSTF were filtered out through ADME screening and 84 putative targets correlated with different diseases. Pathway enrichment analysis showed that the candidate targets were mostly related to the response to bacteria and immunity signalling pathways, which are known contributors to pharyngitis. Experimental results confirmed that RSTF exerted therapeutic effects on pharyngitis mainly by antibacterial effect and downregulation of NF-κB activities. It is demonstrated both in silico and in vivo/vitro that RSTF exerted therapeutic effects on pharyngitis mainly through an antibiotic effect and downregulation of NF-κB signalling pathway.

Study on the effect of regulation of Cordyceps militaris polypeptide on the immune function of mice based on a transcription factor regulatory network.

BACKGROUND: The pathogenesis of the abnormality of the immune system is still not clear at present. Chemosynthetic drugs, human or animal immune products and microbiological drugs are used as the main drugs in clinics currently, but these drugs have different side effects. So researchers turned to safer natural products in order to find immunomodulatory active substances from natural products and their extracts. METHODS: Immunosuppressed mice were induced by cyclophosphamide and administered with Cordyceps militaris polypeptide (CMP) for the study on the effect of CMP on the immune function of mice and its mechanism. Based on the 1748 differential gene sets selected in our previous work, the transcription factors and their corresponding target genes were screened by integrating the TRED (Transcriptional Regulatory Element Database), a transcriptional factor-target gene regulatory network was constructed, then the role of transcription factors in the regulatory network was elucidated by statistically analyzing the key nodes, and finally, the correlation of network genes with diseases was analyzed by using the DAVID database. RESULTS: The results of animal experiments showed that CMP could increase the immune organ indexes, the number of white blood cells, the degree of delayed allergy and the content of hemolysin in the serum of mice. CMP was found to be involved in the regulation of immune function in mice through genes Kdr, Spp1, Ptgs2, Rel, and Smad3, and transcription factors Ets1, E2f2 and E2f1. E2F2 and E2F1 are members of the E2F family, so we speculated that the E2F family might play an important role, and its main regulatory pathways were the PI3K-Akt signaling pathway and TNF signaling pathway. CONCLUSION: CMP can improve the immunity of mice. CMP can regulate the immune function of mice through multiple genes and transcription factors, and may also play a role in immune-related diseases, such as cancer.

Protective effects of Ulva pertusa polysaccharide and polysaccharide­iron (III) complex on cyclophosphamide induced immunosuppression in mice.

A water-soluble polysaccharide was isolated from marine green algae Ulva pertusa and then chelated with iron to prepare the polysaccharide­iron (III) complex. The immunomodulatory activities of sulfated polysaccharide and polysaccharide­iron (III) complex were investigated through a mice immune-deficiency model. Cyclophosphamide (Cy) was utilized to establish mice immunodeficiency model. Both polysaccharide and polysaccharide­iron (III) complex were proved to promote the proliferation of lymphocyte and enhance the activities of mice macrophages. In mice serum, the levels of cytokines including TNF-α, IFN-γ and IL-10 restored and the contents of hemolysin were also found elevated after treatment with polysaccharide and its iron complex. Besides, it has been shown that both polysaccharide and polysaccharide­iron (III) complex increased the contents of Hb, RBC and HCT in mice blood, and the effect of iron complex was better. All these results suggested that Ulva pertusa polysaccharide could be developed as a healthy function food. It was also noteworthy that the polysaccharide­iron (III) complex showed no negative effect upon the immunomodulatory activity of polysaccharide. Instead, the polysaccharide­iron (III) complex showed excellent hematopoietic capacity perhaps due to the supplement of iron.

Glycosaminoglycan from Apostichopus japonicus induces immunomodulatory activity in cyclophosphamide-treated mice and in macrophages.

This study was designed to systematically elucidate the immunomodulation effect of glycosaminoglycan from Apostichopus japonicus (AHG) in cyclophosphamide (CY)-induced immunosuppression model and potential mechanism responsible for the activation of macrophages. The results showed that the treatment with AHG could increase natural killer (NK) cell cytotoxicity, carbon clearance and marker enzymes activities in CY-induced immunosuppression mice, indicating that the innate immunity experienced recovery to some extent. Moreover, CY-induced reductions in thymus and spleen indices, serum levels of cytokines, immunoglobulins and hemolysin, as well as the ratio of spleen lymphocyte subsets were recovered by AHG, suggesting that AHG could improve the adaptive immunity through cellular immunity and humoral immunity. Delightedly, it was found that AHG at 10 mg/kg body weight could restore the CY-induced immunosuppression in mice to normal level on both innate and adaptive immunity. Furthermore, AHG also promoted both the expression of NO, TNF-α, IL-6, IL-1ß, IL-18 and MCP-1 protein and related mRNA in macrophages. It was revealed that AHG activated macrophages through the phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear factor-B (NF-κB). In conclusion, AHG exerts remarkable immunomodulatory activities in both innate and adaptive immune system. These findings should have great value for further study on the immunopotentiating mechanisms of this biomacromolecule.

α-Haemolysin production, as a single factor, causes fulminant sepsis in a model of Escherichia coli-induced bacteraemia.

α-Haemolysin (HlyA) from uropathogenic Escherichia coli has been demonstrated to be a significant virulence factor for ascending urinary tract infections. Once the E. coli reach the well-vascularised kidneys, there is a high risk of bacteraemia and a subsequent septic host response. Despite this, HlyA has the potential to accelerate the host response both directly and via its ability to facilitate adenosine triphosphate release from cells. It has not been settled whether HlyA aggravates bacteraemia into a septic state. To address this, we used an E. coli strain in a model of acute urosepsis that was either transfected with a plasmid containing the full HlyA operon or one with deletion in the HlyA gene. Here, we show that HlyA accelerates the host response to E. coli in the circulation. Mice exposed to HlyA-producing E. coli showed massively increased proinflammatory cytokines, a substantial fall in circulating thrombocytes, extensive haematuria, and intravascular haemolysis. This was not seen in mice exposed to either E. coli that do not secrete HlyA or vehicle controls. Consistent with the massive host response to the bacteria, the mice exposed to HlyA-producing E. coli died exceedingly early, whereas mice exposed to E. coli without HlyA production and vehicle controls survived the entire observation period. These data allow us to conclude that HlyA is a virulence factor that accelerates a state of bacteraemia into fulminant sepsis in a mouse model.

Sea cucumber (Codonopsis pilosula) oligopeptides: immunomodulatory effects based on stimulating Th cells, cytokine secretion and antibody production.

This study aimed to investigate the immunomodulating activity of small molecule oligopeptides from sea cucumber (Codonopsis pilosula) (SOP) in mice. Seven assays were performed to determine the immunomodulatory effects, including splenic lymphocyte proliferation and delayed-type hypersensitivity assays (cell-mediated immunity), IgM antibody response of spleen to sheep red blood cells (SRBC) and serum hemolysin level assays (humoral immunity), the carbon clearance assay and the phagocytic capacity of peritoneal cavity phagocytes assay (macrophage phagocytosis), and the NK cell activity assay. Spleen T lymphocyte subpopulations, multiplex sandwich immunoassays of serum cytokine and immunoglobulin levels and enzyme-linked immunosorbent assays for small intestinal secretory immunoglobulin were performed to study the mechanism by which SOP affects the immune system. We found that SOP could improve immune functions in mice, which may be due to the enhancement of the functions of cell-mediated immunity, humoral immunity, macrophage phagocytosis and NK cell activity. From the cellular and molecular assays, we postulated that the immunomodulatory effects are most likely attributed to the stimulation of Th cells, cytokine secretion and antibody production.

Schisandra polysaccharide evokes immunomodulatory activity through TLR 4-mediated activation of macrophages.

Schisandra chinensis (Turcz.) Baill has been used in traditional Chinese medicine for centuries. Previous studies have shown that Schisandra polysaccharide (SCPP11) has robust antitumor activity in vivo. In this study, the immunomodulatory activity and mechanisms of action of SCPP11 were investigated further to reveal its mechanism of action against tumors. Results showed that SCPP11 increased the thymus and spleen indices, pinocytic activity of peritoneal macrophages, and hemolysin formation in CTX-induced immunosuppressed mice. Moreover, SCPP11 significantly increased immunoglobulin levels, cytokines levels in vivo and induced RAW264.7 cells to secrete cytokines in vitro. RAW264.7 cells pretreated with SCPP11 significantly inhibited the proliferation of HepG-2 cells. In addition, SCPP11 promoted both the expression of iNOS protein and of iNOS and TNF-α mRNA. TLR-4 is a possible receptor for SCPP11-mediated macrophage activation. Therefore, the data suggest that SCPP11 exerted its antitumor activity by improving immune system functions through TLR-4-mediated up-regulation of NO and TNF-α.

Purification, characterization and immunostimulatory activity of polysaccharide from Cipangopaludina chinensis.

In this study, we investigated the purification, preliminary characterization and immunostimulatory activity in vivo of polysaccharide from Cipangopaludina chinensis (CCPS). Firstly, crude CCPS was prepared by hot water extraction. And the crude CCPS was sequentially purified by chromatography of DEAE-52 and Sephadex G-100, resulting in two purified fractions of CCPS-1 and CCPS-2. We found the two fractions were homogeneous heteropolysaccharides mainly composed of rhamnose and glucose with the average molecular weight of 226 and 235 kDa, respectively. CCPS-2 was quite different from CCPS-1. It had much higher content of uronic acid and sulfuric radical. For immunostimulatory activity in vivo, crude CCPS could significantly increase the thymus and spleen indices, enhance the macrophage function, and increase the level of serum hemolysin in cyclophosphamide-treated mice, suggesting CCPS had a potent immunostimulatory activity and could be explored as a potential natural immunomodulatory agent.

Polysaccharide extracted from Rheum tanguticum prevents irradiation-induced immune damage in mice.

AIM: To investigate the protective effect of purified fraction 1 polysaccharide extracted from Rheum tanguticum RTP1 on irradiation-induced immune damage in mice. METHODS: Kunming mice were randomly divided into five groups: normal group (NC), irradiation control group (IC), RTP1 low dose (200 mg/kg), middle dose (400 mg/kg) and high dose (800 mg/kg) groups. RTP1 was administered by the gastric route for 14 d, mice in the NC and IC groups being given by 0.9% sodium chloride solution in the same way. The mice in all groups except NC group were irradiated with 2.0 Gy6°Co γ-ray on the fourteenth day. Immune indives of non-specific immune function, cellular immunity and humoral immunity were assessed at the 24th hour after radiation. RESULTS: Compared with the IC group, the spleen index, thymus index, rate of carbon clearance, phagocytic function of macrophages, lymphocyte proliferation, hemolysin value of blood serum and NK activity were increased markedly (P < 0.05 or P < 0.05). CONCLUSION: RTP1 has an obvious protective effects on damage in γ-ray radiated mice.

Immunomodulatory activity of polysaccharide-protein complex of longan (Dimocarpus longan Lour.) pulp.

The immunomodulatory function of longan pulp polysaccharide-protein complex (LP3) was investigated in immunosuppressed mice models. Compared with the model control, peroral administration of 100 mgkg⁻¹d⁻¹ LP3 could significantly increase/enhance antibody production against chicken red blood cell (CRBC), concanavalin A (ConA)-induced splenocyte proliferation, macrophage phagocytosis, NK cell cytotoxicity against YAC-1 lymphoma cell, and interferon-gamma (INF-γ) and interleukin-2 (IL-2) secretion in serum (P < 0.05). The immunomodulatory effects, except for those on splenocytes and macrophages (P > 0.05), were also observed in mice administered with 50 or 200 mgkg⁻¹d⁻¹ LP3 (P < 0.05). The beneficial effects of 50-200 mgkg⁻¹d⁻¹ LP3 were comparable to those of 50 mgkg⁻¹d⁻¹ ganoderan. The strong immunomodulatory activity of LP3 confirmed its good potential as an immunotherapeutic adjuvant.

Maternal and fetal exposure to pesticides associated to genetically modified foods in Eastern Townships of Quebec, Canada.

Pesticides associated to genetically modified foods (PAGMF), are engineered to tolerate herbicides such as glyphosate (GLYP) and gluphosinate (GLUF) or insecticides such as the bacterial toxin bacillus thuringiensis (Bt). The aim of this study was to evaluate the correlation between maternal and fetal exposure, and to determine exposure levels of GLYP and its metabolite aminomethyl phosphoric acid (AMPA), GLUF and its metabolite 3-methylphosphinicopropionic acid (3-MPPA) and Cry1Ab protein (a Bt toxin) in Eastern Townships of Quebec, Canada. Blood of thirty pregnant women (PW) and thirty-nine nonpregnant women (NPW) were studied. Serum GLYP and GLUF were detected in NPW and not detected in PW. Serum 3-MPPA and CryAb1 toxin were detected in PW, their fetuses and NPW. This is the first study to reveal the presence of circulating PAGMF in women with and without pregnancy, paving the way for a new field in reproductive toxicology including nutrition and utero-placental toxicities.

[Studies on chemistry component and the biological activity of petroleum ether extraction from pre-and post-processed of Cornus officinalis].

OBJECTIVE: To investigate substance basis for improving pharmacodynamics by comparing the chemical constituents of petroleum ether extraction from crude and processed Cornus officinalis and the effects on the immunologic function of mice with immunosuppression induced. METHODS: The volatile components in petroleum ether extraction were analyses by GC-MS. Non-specific immune function was determined by cleaning carbon particle index K, Swallow index a, spleen index and thymus index. The specific humoral immune function was evaluated by detecting the content of serum hemolysin. RESULTS: The chemical constituents of petroleum ether extraction of Cornus officinalis before and after being processed had significant changes. After being processed, Vitamin E increased by 46.6%, linoleic acid increased by 18.3% and methyl linen increased by 30.9%. The extraction increased clearance rate of charcoal carbon particles index K, swallow index alpha, spleen index, thymus index and the level of serum hemolysin. CONCLUSION: The extraction can markedly improve non-specific immune function and the specific humoral immune function which are active sites of improving immunologic function and post-processed is better. Substance basis could be Vitamin E, Linoleic acid, methyl linen and so on.

[Protecting effect of Chaenomeles speciosa broth on immunosuppressive mice induced by cyclophosphamide].

OBJECTIVE: To investigate the effects of Chaenomeles speciosa broth on immunoregulation for anti-tumor chemotherapy. METHODS: Immunosuppressive model was induced by cyclophosphamide (CTX) in mice. The mice were treated with the broth for 15 days. The serum hemolysin was observed in mouse sera. Spleen lymphocyte transformation and gene transcription related to the immunoregulation in spleen lymphocytes were detected. RESULTS: After administrated the broth, the serum hemolysin and lymphocyte transformation rates significantly increased and the mRNA expression of foxp3, TGF-beta, PD1, Fas, Bax were downregulated compared with CTX-group. CONCLUSION: Chaenomeles speciosa broth has protective effects on the immunosuppressive mouse induce by CTX.

Antioxidant and immunity activity of water extract and crude polysaccharide from Ficus carica L. fruit.

The antioxidative activities of water extract (WE) and crude hot-water soluble polysaccharide (PS) from Ficus carica L. fruit were investigated using various assays in vitro, including scavenging abilities on DPPH, superoxide and hydroxyl radicals and reducing power. The immunity activities of PS were evaluated using the carbon clearance test and serum hemolysin analysis in mice. In addition, total phenolics and flavonoids contents were also determined. Both WE and PS have notable scavenging activities on DPPH with the EC(50) values of 0.72 and 0.61 mg/ml, respectively. The PS showed higher scavenging activity than WE on superoxide radical (EC(50), 0.95 mg/ml) and hydroxyl anion radical (scavenging rate 43.4% at concentration of 4 mg/ml). The PS (500 mg/kg) also has a significant increase in the clearance rate of carbon particles and serum hemolysin level of normal mice. The results indicate that both WE and PS might be applicable in healthy medicine and food industry.

Oral administration of live Bifidobacterium substrains isolated from healthy centenarians enhanced immune function in BALB/c mice.

Generally, there is an age-related decline in the human gut titer of Bifidobacterium species, but the titer in healthy centenarians was previously reported to be comparable to that found in much younger people. We addressed whether elevated Bifidobacterium titers relate positively to immune function. This study evaluated the immunoactivities of 2 Bifidobacterium strains (B adolescentis BBMN23 and B longum BBMN68) isolated from healthy centenarians in China. Different dosages (2 x 10(11), 2 x 10(9), or 2 x 10(7) colony-forming units [CFU]/kg body weight) of live bifidobacteria were orally administered once per day to healthy BALB/c mice, and the control group was given sterile skim milk every day. After 4 weeks, the immune parameters including cellular immunity (delayed-type hypersensitivity [DTH], and splenic lymphocyte proliferation), humoral immunity (serum hemolytic activity in immunized animals), and nonspecific immunity (peritoneal macrophages phagocytsis natural killer [NK] cell activity) were measured. We report that both Bifidobacterium strains independently increased the DTH response. Macrophage phagocytosis was also enhanced, while activities of the NK cells and levels of the serum hemolysin also were significantly higher than in the control group. There was a significant increase in splenic lymphocyte proliferation in bifidobacteria treatment animals compared to controls. In conclusion, ingestion of B. adolescentis BBMN23 and B. longum BBMN68 can enhance both innate and acquired immunity in healthy specific pathogen-free (SPF) mice and strains of bifidobacteria from healthy centenarians in Bama longevity villages in China may possess potentially valuable immunomodulatory properties.

[Study on the effect of Part III from Cynomorium songaricum on immunosuppressive mice induced by cyclophosphamide].

OBJECTIVE: To study the immunomodulatory effect of Part III of Cynomorium songaricum Rupr. on the immunosuppressive mice. METHODS: The immunity-deficiency model was induced by intraperitoneal injection of cyclophosphamide (CTX) at the dose of 100 mg/kg in mice; all the animals were divided into normal control group, immunity-deficiency model group, Part III treated group (300 mg/kg) and positive control group (TSPG, 300 mg/kg). The hemogram of peripheral blood, the index of immune organs, the phagocytosis activity of macrophage, the content of serum hemolysin were measured. RESULTS: The index of organs, the phagocytosis activity of macrophage and the content of serum hemolysin in the model group increased after administrated of Part III. CONCLUSION: Part III from Cynomorium songaricum Rupr. has protective effect on the immunosuppressive mice, which may be related to the increasing of humoral immunity and nonspecific immunity.

Streptococcus agalactiae CAMP factor binds to GPI-anchored proteins.

CAMP factor (protein B) is a pore-forming protein secreted by Streptococcus agalactiae. It causes lysis of sheep red blood cells when these have been sensitized with staphylococcal sphingomyelinase. We here show that CAMP factor binds to GPI-anchored proteins, and that this interaction involves the carbohydrate core of the GPI-anchor. Enzymatic cleavage of GPI-anchors with phosphatidylinositol-specific phospholipase C strongly reduces the sensitivity of erythrocytes to CAMP factor. Incorporation of alkaline phosphatase, a model GPI-anchored protein, into liposome membranes renders the latter susceptible to permeabilization by CAMP factor. GPI-anchored proteins therefore function as cellular receptors for CAMP factor.

[Gastric cytomedines effects on immune response in acute immobilization stress in rats].

Effects of gastric cytomedines on repair and immune response were studied in 137 white non-inbred rats. Erosions of gastric mucosa were induced by acute immobilization stress. A decrease in antibody production, titers of hemagglutinins and hemolysins was found on day 2 after stress. Gastric cytomedines in a dose 0.15 mg/kg intramuscularly stimulate immune response (increased production of antibodies, antibody dependent cellular cytotoxicity index, titers of hemagglutinins and hemolysins), fast erosions epithelization and have a protective action.

[Effect of taurochenodeoxycholic acid on immune function in mice].

OBJECTIVE: To observe the influence of Taurochenodeoxycholic Acid (TCDCA) on immun, function in mice. METHODS: T-Cell subgroups were determined by using Flow cytometry; The content of anti-body in serum was assayed by using Spectrophotometry; The phagocytosis of mononuclear phagocyte system was determined by using Carbon particle clearance test and anti-sheep blood cell hemolysin was determined by using Turbidimetric method. RESULTS: TCDCA signifeantly enhanced the percentage of CD and CD19+ lymphocytes and CD4+/CD8+ value in peripheral blood and the content of serum hemolysin and lysozymem in mice. Moreover, TCDCA markedly improved the phagocytosis functions of mononuclear phagocyte system and observably inhibited delayed type hypersensitivity. CONCLUSION: TCDCA can significantly enhance the immune function in mice.