Fisetin alleviates sepsis-induced multiple organ dysfunction in mice via inhibiting p38 MAPK/MK2 signaling.

Sepsis-induced multiple organ dysfunction and inflammatory response are life-threatening symptoms without effective treatment. Fisetin, a dietary flavonoid extracted from berries and family Fabaceae, has displayed neuroprotective and anti-oxidant activities. In this study we investigated whether fisetin exerted a protective effect against sepsis-induced multiple organ dysfunction in mouse cecum ligation and puncture (CLP) model. The mice were injected with fisetin (10 mg/kg, ip) 0.5 h prior to CLP, and sacrificed 18 h after CLP. We found that fisetin administration significantly alleviated CLP-induced lung, liver and kidney injury, as well as the expression levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α and IL-1ß in bronchoalveolar lavage fluid (BALF). In lipopolysaccharide (LPS)-treated mouse bone marrow-derived macrophages (BMDMs), application of fisetin (3-10 µM) dose-dependently inhibited the expression levels of IL-6, TNF-α, IL-1ß, and inducible nitric oxide synthase (iNOS). Furthermore, fisetin dose-dependently inhibited the phosphorylation of p38 MAPK, MK2, and transforming growth factor-ß-activated kinase (TAK) 1 via attenuating the interaction between TAK1 and TAK-binding proteins (TAB) 1. These results demonstrate that fisetin is a promising agent for protecting against sepsis-induced inflammatory response and organ injury via inhibiting macrophage activation.

Expression and clinical significance of Sirt1 in colorectal cancer.

The objective of the present study was to examine the expression of Silent information regulator 1 (Sirt1) in colorectal cancer and peritumoral normal mucosa tissue, and therefore analyze the role and molecular mechanism of Sirt1 in the pathogenesis of colorectal cancer. Colorectal cancer tissue specimens were employed as the experimental group, and adjacent normal mucosa tissues >5 cm from tumor lesions were used as the control group. The expression of Sirt1 was detected by the immunohistochemical streptavidin peroxidase detection method in paraffin-embedded sections, whilst Sirt1 protein expression was examined by western blot analysis in the fresh tissues. Sirt1 protein was primarily expressed in the nuclei of the tumor cells, and positive staining was brownish-yellow in color. The relative expression quantities of Sirt1 in the peritumoral normal rectal mucosa and rectal carcinoma were 1.15 and 2.62, and the differences between the two groups were statistically significant (P<0.05). The expression level of Sirt1 in colorectal carcinoma was significantly associated with the depth of tumor invasion, differentiation and tumor size (P<0.05). Sirt1 expression was also found to be associated with tumor tissue type, lymph node metastasis, Duke's stage and patient age. These characteristics combined may therefore be used as markers for the early diagnosis of colorectal cancer pathogenesis.

[Effects of serums containing Buzhong Yiqi decoction with Astragalus Radix or Hedysari Radix on anti-immunosenescence in spleen lymphocytes of SAMP8 mice].

This paper was aimed to compare the effect of Buzhong Yiqi decoction containing Hedysari Radix or Astragali Radix on anti-immunosenescence effects in spleen lymphocytes of senescence accelerated mouse 8 (SAMP8). The effect of the serums on the proliferation of spleen T lymphocytes in SAMP8 mice induced by ConA was tested by MTT. The effect of the serums on the T lymphocytes subsets of SAMP8 mice was measured by flow cytometry. ELISA was used to detect the level of IL-2 and IFN-γ in the culture supernatants of spleen lymphocytes. The effect of the serums on the expression of CD28 mRNA in spleen T lymphocytes was detected by fluorescent quantitative PCR. Western blot was used to detect the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. Both the serums of Buzhong Yiqi decoctions containing Hedysari Radix or Astragali Radix improved the proliferation of T lymphocytes in SAMP8 mice. Both the serums had no obvious effect on the differentiation of spleen T lymphocytes'subsets in SAMP8 mice. Both the serums increased the content of IL-2 and INF-γ in the culture supernatants of spleen lymphocytes. And for the content of IL-2, the serum of Buzhong Yiqi decoction with Hedysari Radix was better（P<0.05）. Both the serums improved the expression of CD28 mRNA in spleen T lymphocytes of SAMP8 mice. And the effect of Hedysari Radix group was better than that of Astragalus Radix group（P<0.05）. Both the serums improved the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. The role of the serums containing Buzhong Yiqi decoction with Astragalus Radix and the decoction with Hedysari Radix in anti-immunosenescence was through the effect of the CD28. And the effect of Hedysari Radix group was better than that of Astragalus Radix group on improved the expression of CD28 mRNA in T lymphocytes of SAMP8 mice. Astragalus Radix and Hedysari Radix could swap in the aspect of anti-immunosenescence.

[Intervention of Qi-activating and Spleen-strengthening Herbs on Ca2+/CaMK II Signaling Pathways Key Factors in Skeletal Muscle Tissue of Rats with Spleen-qi Deficiency].

OBJECTIVE: To observe changes of [Ca2+]i concentration and CaM, CaMK II and p-CaMK II of Ca2+/CaMK II signaling pathways in skeletal muscle tissue of rats with spleen-qi deficiency and intervention of Sijunzi decoction and extract of Hedysarum polybotrys. METHODS: Rats were randomized into four groups: normal control group, spleen-qi deficient model group, extract from Hedysarum polybotrys group and Sijunzi decoction group, ten rats in each group. After the spleen-qi deficient models were built by comprehensive application of rhubarb, exhaustive and hungry methods, and treatment groups were treated with extract from Hedysarum polybotrys at 6 g/(kg . d) or Sijunzi decoction at 20 g/(kg . d) for 21 d. Then, general existence,gastrointestinal hormones GAS and MOT levels, and activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase of skeletal muscle were evaluated. Also, confocal laser technology was used to test cellular[Ca2+]i concentrations in skeletal muscle and Western blotting technique was used to test CaM, CaMK II and p-CaMK 11 expression in intestinal tissue of spleen-qi deficient model rats. RESULTS: Compared with normal group, general condition was poor, levels of GAS and MOT decreased (P <0. 01), activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase, [Ca2+]i concentration as well as expression of CaM, CaMK II and p-CaMK II in skeletal muscle decreased significantly (P < 0. 01) in spleen-qi deficienct model rats. Compared with model group, general condition improved significantly, as well as level of MOT in intestinal increased (P <0. 05) in the rats of extract from Hedysarum polybotrys group and Sijunzi decoction group,while level of GAS increased in intestinal(P <0. 05) in the rats of Sijunzi decoction group; Moreover, activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase as well as [Ca2+]i concentration and expression of CaM and CaMK II in skeletal muscle tissue increased (P < 0. 05) in the rats of extract from Hedysarum polybotrys group and Sijunzi decoction group, while p-CaMK II in skeletal muscle tissue increased in the rats of Sijunzi decoction group (P < 0. 05). CONCLUSION: Sijunzi decoction and extract of Hedysarum polybotrys can be applied to treat spleen-qi deficiency syndrome through the mechanism of regulating GAS and MOT secretion and raising expression of Ca2+ /CaM signaling pathways key factors in skeletal muscle tissue. Sijunzi decoction has the better effect

Radix Hedysari polysaccharide suppresses lipid metabolism dysfunction in a rat model of non­alcoholic fatty liver disease via adenosine monophosphate­activated protein kinase pathway activation.

Oxidative stress and excess hepatic lipid accumulation contribute to non­alcoholic fatty liver disease. Radix Hedysari polysaccharides (RHP) have attracted interest due to their antioxidant properties and immunomodulatory effects. However, the effect of RHP on hepatic lipid metabolism remains to be elucidated. In the present study, the response of Sprague­Dawley rat livers to a high­fat diet and RHP treatment was investigated by evaluating body weight, liver histology, hepatic lipid content, adenosine monophosphate­activated protein kinase (AMPK) activity and lipid metabolism gene transcriptional profiles. The present study demonstrated that RHP ameliorated lipid metabolism disorders, regulated hepatic lipid content, improved liver inflammation and damage, activated AMPK via phosphorylation, upregulated peroxisome proliferator­activated receptor α and downregulated the mRNA expression of sterol regulatory element binding protein­1c in rat livers, which reduced lipogenesis and increased lipolysis. Taken together, these results suggested that RHP effectively ameliorates lipid metabolism disorders in rat livers; thus, RHP may be a potential therapeutic agent in the prevention of hepatic steatosis.

[Comparative study on substituting hedyseri radix for astragali radix in serum containing yiqiyangxue prescription on aged mice spleen lymphocyte proliferation and anti-oxidant effect].

OBJECTIVE: By substituting Hedyseri Radix for Astragali Radix in Yiqiyangxue prescription, to compare the effects of both serum containing medicine on aged mice spleen lymphocyte proliferation and anti-oxidant effect. METHOD: After using the same dose of Hedyseri Radix to replace Astragali Radix in Yiqiyangxue prescription, the best concentration of serum containing medicine,the best incubation time and the effects of ConA-induced spleen lymphocyte proliferation were determined by MTY method. Use reagent kits to detect the activity of SOD, MDA and ROS levels in aged mice spleen lymphocytes and IL-2 level in culture supernatant fluid of spleen lymphocytes. RESULTS: Both serum containing medicine can enhance the proliferation of aged mice spleen lymphocytes. The best concentration of serum containing medicine was 40% and the incubation time was 72 h. The serum containing Yiqiyangxue of Hedyseri Radix prescription acted more effective than that of Astragali Radix on the enhancement of proliferation. Both serum containing medicine showed similar effects on increasing SOD activity, IL-2 level and decreasing MDA and ROS level. Moreover,serum of Hedyseri Radix was superior in the enhancement of proliferation, IL-2 and the reduction of ROS level. CONCLUSION: Both serum containing medicine of Hedyseri Radix and Astragali Radix generate the same effect of anti-aging and enhancement of proliferation.

[Comparative study of Hedysari Radix and Astragali Radix alternative classic tonification prescriptions on humoral immunity in immunosuppressed mice].

OBJECTIVE: To compare the regulating effects of Hedysari Radix and Astragali Radix alternative classic tonification prescriptions on humoral immunity in immunosuppressed mice. METHODS: The immunosuppressed mouse model was induced by cyclophosphamide. The mice were administered intragastically with same dose of Hedysari Radix and Astragali Radix alternative Buzhong Yiqi Yiqi Yangxue,Yupingfeng oral liquid and Fuqi Zhihan granules for antagonistic experiments in vivo. And spleen index, HC50, CD19+B lymphocyte subgroup and content of serum IL-4 were determined after treatment. RESULTS: Both groups of Hedyseri Radix and Astragali Radix could antagonize immunosuppressive action caused by cyclophosphamide. They both could significantly raise spleen index, HC50, CD19+ B lymphocyte subgroup and content of serum IL4 in different degree. And Yupingfeng aqueous extract of Hedysari Radix substitute Astragali Radix was better than Yupingfeng oral liquid in raising spleen index. There were no significant differences among the rest Hedysari Radix and Astragali Radix alternative groups. CONCLUSION: Hedysari Radix compatibility with other drugs compared with original prescription has similar role in humoral immunity regulation.

[Proteomics analysis of the synergy effect of Radix Hedysari polysaccharides combined with chemotherapy on S180 tumor cells].

OBJECTIVE: To analyze the differentially expressed proteins of the synergy effect of Radix Hedysari polysaccharides (HPS)combined with chemotherapy (Cy) on S180 tumor cells. METHODS: The total proteins extracted from the HPS combined with Cy treated S180 cells in tumor-bearing mice were separated by two dimentional gel electrophoresis (2-DE)and compared with those from Cy treated S180 cells using PDQuest 8.0 software. Mass spectrometry was applied to identify the differentially expressed proteins. Western blot was used to determine the differential expression of one protein. RESULTS: Twenty-four differentially-expressed proteins in HPS group were discovered. The five differential expressed proteins among twenty-four proteins were later identified by mass spectrometry and Mascot software as heat-shock protein hsp84 (HSP90beta), apolipoprotein A, albumin, heat shock protein beta-1 (HSP27)and unnamed protein product, including one up regulated and four down regulated expressed proteins respectively. Results from Western blot manifested the same trend as from proteomics analysis. CONCLUSION: Proteomics technique can be used to discover target proteins associated with the synergy effect of HPS combined with Cy on S180 tumor cells, involving some important proteins related to energy metabolism, oxidative stress and apoptosis induction signal transduction.

Proteomic analysis of the effect of triterpenes from Patrinia heterophylla on leukemia K562 cells.

For centuries, Patrinia heterophylla had been used in China to treat many diseases including tumor. Triterpenes has been identified as the major active constituents in Patrinia heterophylla. To elucidate the antitumor mechanism of triterpenes from Patrinia heterophylla1 (TPH), a proteomic analysis is carried out with TPH treatment in K562 cells. The total proteins extracted from TPH treated K562 cells are analyzed by two dimensional gel electrophoresis (2-DE) and compared with those untreated K562 cells. Mass spectrometry is applied to identify the differentially expressed proteins. Twenty-three differentially expressed significant proteins are discovered. Eight proteins are later identified by mass spectrometry (MALDI-TOF-MS) and Mascot software. Among them, four proteins are up-regulated (Aldolase A, Glyceraldehyde-3-phosphate dehydrogenase, Flavin reductase and Hemoglobin subunit) and four proteins were down-regulated (Heat-shock protein 90 〈Alpha〉 (HSP90-〈Alpha〉), Eukaryotic translation initiation factor 5A, Moesin, tublin) by TPH treatment in K562 cells. The identified proteins are associated with energy metabolism, oxidative stress, apoptosis, signal transduction, differential induction, and protein biosynthesis. These findings might provide valuable insights into the antitumor mechanism of TPH in K562 cells.

[Comparative study of Radix Hedyseri as sulstitute for Radix Astragali of yupingfeng oral liquid on cellular immunity in immunosuppressed mice].

OBJECTIVE: To study on Radix Hedyseri as substitute for Radix Astragali of Yupingfeng oral liquid on cellular immunity in immunosuppressed mice. METHODS: The model of immunosuppression mice were induced by Cyclophosphamide. And the same dose of Radix Hedyseri and Radix Astragali alternative Yupingfeng oral liquid was intragastric administrated into mice; Antagonistic experiments were observed in vivo. Determined the thymus gland index, spleen index, phagocytosis of the macrophage, proliferation index of T lymphocyte, kill and wound activity, T lymphocyte subgroup, and content of IL-13 of serum. RESULTS: Yupingfeng oral liquid and Yuping-feng aqueous extract of Radix Hedyseri substitute Radix Astragali both could significant raise thymus gland index and spleen index, and clearly increase the phagocytosis of the macrophage. They both could antagonize immunosuppressive action caused by Cyclophosphamide, which could promote T lymphocyte proliferation, kill and wound activity, quantity of T lymphocyte subgroup, and production of IL-1beta with different degree. And Yupingfeng aqueous extract of Radix Hedyseri substitute Radix Astragali increased spleen index and T lymphocyte proliferation was better than those of Yupingfeng oral liquid. CONCLUSION: Radix Hedyseri and Radix Saposhnikoviae, compatible with Rhizoma Atractylodis Macrocephalae compared with Yupingfeng oral liquid in cell immunity regulation has a similar role, and better in the recovery of spleen weight and T cell proliferation.

WITHDRAWN: Effect of phytoestrogen isoflavone on MPP(+)-induced apoptosis in PC12 cells.

The Publisher regrets that this article is an accidental duplication of an article that has already been published, http://dx.doi.org/10.1016/j.bionut.2010.09.004. The duplicate article has therefore been withdrawn.

[Response of applying palygorskite and fertilization on growth rules of Radix Hedysari].

OBJECTIVE: Study on the growth development and fertilizer requirement regularity of Radix Hedysari. METHODS: Adopted the field experiment, investigate the influence of Radix Hedysari by applying different amount of palygorskite and fertilization. RESULTS: The growth process of plant height and branch height of Radix Hedysari were divided into two stages, the stage from emergence to late July was fast growth phase, and the stage from late July was slow growth stage. The dry matter accumulation center of Radix Hedysari transferred from aerial part to underground part in late July. Single application of palygorskite (1500 kg/hm2) and single application of palygorskite (2250 kg/hm2) increased plant height, promoted dry matter accumulation in aerial part and root of Radix Hedysari. Combined application of palygorskite (1500 kg/hm2) and NPK fertilizer and combined application of palygorskite (2250 kg/hm2) and NPK fertilizer improved growth indexes of Radix Hedysari compared with single application of NPK fertilize. CONCLUSION: The yield of Radix Hedysari was improved by applying palygorskite and fertilization.

[Effects of extracts from Patrinia heterophylla on gene expression patterns during morphogenesis of chicken limb buds in vivo].

OBJECTIVE: To study the effects of the extracts from Patrinia heterophylla on gene expression patterns during morphogenesis of chicken limb buds in vivo. METHODS: Implanted a bead into an chicken embryo, which was soaked in the extracts from Patrinia heterophylla. Detected the extracts-induced morphogenesis changes (Myf5, Myod and PCNA). RESULTS: The extracts from Patrinia heterophylla (200 mg/mL) could affect limb bud development, reduce gene expression of MyfS, MyoD and PCNA. CONCLUSION: The extracts from Patrinia heterophylla can inhibit cell differentiation and proliferation.

Does estrogen receptor gene polymorphism play a role in Parkinson's disease?

This study was designed to investigate the frequency of estrogen receptor (ER) gene polymorphism in Chinese patients with Parkinson's disease (PD). Polymerase chain reaction (PCR) method and restriction fragment length polymorphism (RFLP) were used to detect the ER gene polymorphisms in 158 PD patients and 146 healthy controls. In the PD and control groups, "x" accounted for 83.5% and 80.8%, respectively (P>0.05). "xx" was found in 77.2% of the PD group and in 69.9% of the control group (P>0.05). The frequency of "p" in the PD and control group was 67.7% and 64.0%, respectively (P>0.05). "pp" was 51.9% in the PD group and 43.8% in the control group (P>0.05). "ppxx" was found in 49.4% of the PD and 43.0% of the control subjects (P>0.05). There was no significant difference in the "x", "xx", "p", "pp" or "ppxx" between males and females within the PD or control groups. In conclusion, we found no significant differences in the genotype or allele frequencies between patients with Parkinson's disease and healthy subjects. These findings suggest that the estrogen receptor gene polymorphism may not play a key role in the pathogenesis PD in Chinese patients.

Protective effect of estrogen on apoptosis in a cell culture model of Parkinson's disease.

OBJECTIVES: The protective effect of estrogen on the neurons in Parkinson's disease (PD) is unclear. The present study aimed to investigate the effect of estrogen on the apoptosis and dopaminergic function on a cultured cell model of PD. METHODS: The PD model was established by addition of 1-methyl-4-phenylpyridinium (MPP+) to PC12 cell culture. Estrogen was added to cell groups with MPP+ (Estrogen+MPP+), and without MPP+ (Estrogen only group). Cell viability, content of tyrosine hydroxylase (TH), apoptosis ratio, expression of apoptosis-suppression protein Bcl-x and apoptosis-acceleration protein IL-1 beta converting enzyme (ICE) were measured. RESULTS: Cell viability in the Estrogen+MPP+ group was similar to the control group but was higher than in the MPP+ group (P < 0.05). The apoptosis ratios in the Estrogen+MPP+ group (33.6%), and the control group (31.3%), were also similar, but it was lower than in the MPP+ group (63.5%, P < 0.05). Concentrations of Bcl-x were higher in the Estrogen+MPP+ group, whereas ICE concentrations were lower than in the MPP+ group (P < 0.05). CONCLUSIONS: Estrogen suppresses apoptosis and improves cell viability in MPP+ induced injuries in the PC12 cells. The beneficial effects of estrogen on the PD model are due to the suppression of pro-apoptotic protein ICE, and stimulation of anti-apoptotic protein Bcl-x.

[Effects of Patrinia scabra bunge macroporous adsorptive resins extracts on erythrocyte immune function in tumor bearing mice].

OBJECTIVE: To study the erythrocyte immuno-regulatory effect of Patrinia scabra Bunge extracts extracted by macroporous adsorptive resins in tumor bearing mice. METHODS: Patrinia scabra Bunge was extracted by macroporous adsorptive resins, and the amount of polysaccharides and saponins in the extract were determined. Mice bearing S180 tumor were treated with the extract and their survival prolongation rate, erythrocyte rosette formation rates of C3b receptor (ERR-CR), immune complex (ERR-IC) and tumor cell (ERR-TC), as well as the CD35 and CD44s were observed. RESULTS: Polysaccharide content was 21.4%, saponin 41.8% in the extract. As compared with the model group, the survival rate was increased, the erythrocyte immune function was improved (showed increase of ERR-CR and ERR-TC, decrease of ERR-IC), and the amount of CD35 and CD44s in red blood cell membrane increased in mice after being treated with the extract (P < 0.05 or P < 0.01). CONCLUSION: Extract of Patrinia scabra Bunge extracted by macroporous adsorptive resins can regulate the erythrocyte immune function to a certain extent.

[Studies on extraction process optimization of patrinia scabra Bunge saponins].

OBJECTIVE: To optimize the conditions for the extraction of Patrinia scabra Bunge saponins. METHODS: Orthogonal experimental design and ultrasonic method were employed to examine the conditions for the extraction by determination of saponins. RESULTS: The optimun condition for the extraction of Patrinia scabra Bunge saponins was as follows: 65% ethanol for 40 minutes, 55 degrees C and 210 watt of ultrasonic efficinecy. CONCLUSION: The extraction method of Patrinia scabra Bunge sponins is simple and efficient.

[Erythrocyte immunologic function effects of Patrinia scabra extracts by macroporous adsorptive resins on mice burdened transplanted tumor].

OBJECTIVE: To research the erythrocyte immunoregulation effects of Patrinia scabra extracts by macroporous adsorptive resins on mice burdened transplanted tumor. METHODS: Extracts of Patrinia scabra Bunge were separated by macroporous adsorptive resins, ingredients were analysised. Mice burdened transplanted tumor were given extracted drugs. Life prolongation rate was observed, erythrocyte immunologic function and the CD35, CD44s contents of red blood cell were evaluated. RESULTS: Polysaccharide and saponin accounted for 8.4% and 48.4%. Extracts could porolong life expectancy of mice, improve erythrocyte immunolgic function and increase the CD35 and CD44s contents of red blood cell. CONCLUSION: Extracts of Patrinia scabra Bunge by macroporous adsorptive resins have erythrocyte immunoregulation effects on mice burdened transplanted tumor.

[Effects of Fuzheng Yiliu Granule-medicated serum on apoptosis of liver cancer cells from mice and its mechanism].

OBJECTIVE: To study the effects of Fuzheng Yiliu Granule (FZYLG)-medicated serum on apoptosis of liver cancer cells H22 from mice and its mechanism. METHODS: Liver cancer cells H22 from mice were incubated in culture media containing sera from rabbits medicated with different doses of FZYLG. Flow cytometry was used to examine the cell cycle and analyze the apoptotic rate of the H22 cells. The morphological changes of the H22 cells were observed by transmission electron microscope and the apoptosis related proteins Bcl-2 and Bax were examined by streptavidin-biotin peroxidase complex (SABC) method. RESULTS: FZYLG-medicated serum could influence the cell cycle and stop the proliferation of H22 cells at the G(1)/G(0) phase with apoptotic peak being detected. In culture media with FZYLG-medicated sera, the expression of Bcl-2 decreased while that of Bax increased as compared with that in culture medium with non-medicated serum (P<0.05). CONCLUSION: FZYLG-medicated serum can induce apoptosis of the liver cancer cells H22 by influencing the cell cycle, down-regulating the expression of Bcl-2 and up-regulating the expression of Bax.