Research Note: A novel method for preparation of egg yolk immunoglobulin Y against Porphyromonas gingivalis.

Porphyromonas gingivalis (P. gingivalis, P. g) is the main pathogen of periodontal disease, which is treated with egg yolk immunoglobulin Y (IgY) against P. gingivalis. In order to quickly obtain IgY, 30 hens were immunized with inactivated P. gingivalis. The purification of IgY was carried out by the oleic acid (OA) method and the classical method (AS), respectively. The IgY antibody characteristics and antibacterial effects in HPDLF cells were detected by SDS-PAGE, indirect ELISA, Western blot and viability/toxicity assays. SDS-PAGE and Western blot analysis showed that IgY molecules which were rapidly purified by OA method were complete and specific to P. gingivalis. In addition, the results of crystal violet staining and bacterial staining indicated that IgY could agglutinate with P. gingivalis, inhibiting bacterial invasion of host cells. This study is the first to rapidly and efficiently purify IgY by OA method, and the purified IgY is expected to be used in the detection and treatment of P. gingivalis.

Protective effects and potential underlying mechanisms of sodium copper chlorophyllin against ethanol-induced gastric ulcer in mice.

In study, we aimed to determine the mechanisms underlying the gastroprotective effects of sodium copper chlorophyllin (SCC) against ethanol-induced gastric ulcer injury in mice. First, the gastroprotective effects of SCC against gastric ulcer induced by ethanol were assessed. Then, biochemical, histopathological, immunohistochemistry assays, and western blot analysis were conducted to determine the possible mechanisms of action underlying the effects of SCC. Compared to the effects of omeprazole (OME) in a confirmed mouse model of ethanol-induced gastric ulcer injury, treatment with various doses of SCC resulted in up-regulation of Bcl-2 and down-regulation of the pro-apoptotic protein Bax. Significant decreases in the levels of the malondialdehyde (MDA), myeloperoxidase (MPO), and NO in the gastric tissues were observed. Furthermore, inflammatory cytokine analysis revealed that SCC treatment inhibited the expressions of TNF-α and IL-6, greatly reduced the phosphorylation level of IκB, and repressed the nuclear translocation of NF-κB p65, which demonstrated that SCC inhibited the activation of the NF-κB pathway. The present findings suggest that the protective effects of SCC may be beneficial as a potential preventive and therapeutic agent for gastric ulcer through the NF-κB pathway. Taken together, SCC administration significantly decreased the levels of MPO, NO, and MDA in gastric tissue and exerted a powerful anti-inflammatory activity as demonstrated by reduction in the secretions of proinflammatory mediators such as IL-6 and TNF-α in the serum of mice exposed to ethanol.

Bioinformatic analysis of miR-4792 regulates Radix Tetrastigma hemsleyani flavone to inhibit proliferation, invasion, and induce apoptosis of A549 cells.

BACKGROUND: Radix Tetrastigma hemsleyani, a kind of Chinese medicinal herb, contains multiple medicinal ingredients and can exert a variety of pharmacological activities. Our previous study revealed that miR-4792 was significantly upregulated in Radix Tetrastigma hemsleyani flavone (RTHF)-treated A549 cells; however, the regulatory mechanism of RTHF-treated A549 cells remains unclear. MATERIALS AND METHODS: In this study, we investigated the antitumor mechanism and regulatory pathway of miR-4792 in RTHF-treated A549 cells, and the target genes were predicted and pathway enrichment of miR-4792 was performed using bioinformatic analysis. RESULTS: Our results confirmed that the upregulated expression of miR-4792 could inhibit cell proliferation and invasion, provoke cell cycle arrest, and induce apoptosis in A549 cells. Gene Ontology analysis showed that target genes of miR-4792 were enriched in protein binding, cytosol, cytoplasm, plasma membrane, and metal ion binding. Kyoto Encyclopedia of Genes and Genomes analysis showed that target genes of miR-4792 were enriched in aminoacyltRNA biosynthesis, AGE-RAGE signaling pathway in diabetic complications, sphingolipid signaling pathway, neuroactive ligand-receptor interaction, glycosaminoglycan degradation, and regulation of lipolysis in adipocytes. Additionally, FOXC1 was identified as an important target gene of miR-4792 in RTHF-treated A549 cells, and miR-4792 may be the target of some apoptotic-related proteins involved in induction of apoptosis in A549 cells by RTHF. Moreover, the intracellular Ca2+ levels of A549 cells were increased after RTHF treatment, which may be involved in the anticancer regulatory process of miR-4792 in RTHF-treated A549 cells. CONCLUSION: These findings suggest a novel therapeutic approach for lung cancer that will be investigated in future studies.

Analysis of change in microRNA expression profiles of lung cancer A549 cells treated with Radix tetrastigma hemsleyani flavonoids.

BACKGROUND: The aim of this study was to determine the inhibition effects of Radix tetrastigma hemsleyani (RTH) flavonoids on human lung adenocarcinoma A549 cells and the underlying molecular mechanism. RTH is an important Chinese traditional herb that has been widely used in cancer therapy. As an important type of active substance, RTH flavones (RTHF) have been shown to have good antiproliferative effects on various cancer cells. MicroRNAs (miRNAs) are small, noncoding RNA molecules that play important roles in cancer progression and prevention. However, the miRNA profile of RTHF-treated A549 cells has not yet been studied. MATERIALS AND METHODS: The miRNA expression profile changes of A549 cell treated with RTHF were determined using the miRNA-seq analysis. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of differentially expressed miRNAs' (DE-miRNAs) target genes were carried out. RESULTS: In this study, we identified 162 miRNAs that displayed expression changes >1.2-fold in RTHF-treated A549 cells. GO analysis results showed that target genes of DE-miRNAs were significantly enriched in protein binding, binding, cell, cell part, intracellular, cellular process, single-organism process, and single-organism cellular process. Pathway analysis illustrated that target genes of DE-miRNAs are mainly involved in endocytosis, axon guidance, lysosome, melanogenesis, and acute myeloid leukemia pathway. CONCLUSION: These results may assist in the better understanding of the anticancer effects of RTHF in A549 cells.

Radix Tetrastigma hemsleyani flavone inhibits proliferation, migration, and invasion of human lung carcinoma A549 cells.

Radix Tetrastigma hemsleyani flavone (RTHF) is widely used as a traditional herb and has detoxification and anti-inflammatory effects. In this study, we investigated the potential effects of RTHF on the growth and metastasis of human lung adenocarcinoma A549 cells and evaluated its mechanisms. A549 cells were treated with RTHF at various concentrations for different periods. In vitro Cell Counting Kit-8 assay and colony formation methods showed that RTHF had dose- and time-dependent antiproliferation effects on A549 cells. A cell adhesion assay showed that RTHF decreased A549 cell adhesion in a dose-dependent manner. Cell invasion and migration were investigated using the Transwell assay and observed using an inverted microscope; the results showed that cell metastasis was significantly lower in the treatment group than that in the control group (P<0.01). Expression of metastasis-related matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) was detected by real-time polymerase chain reaction and Western blotting. The results showed that the expression of MMP-2, MMP-9, and TIMP-1 decreased, while that of TIMP-2 increased significantly in the RTHF group when compared with the results of the control group. These results show that RTHF exhibits antigrowth and antimetastasis activity in lung cancer A549 cells by decreasing the expression of MMP-2/-9 and TIMP-1 and increasing that of TIMP-2.

[Effect of extract of radix tetrastigma hemsleyani on apoptosis of human lung carcinoma H1299 cells and its mechanism study].

OBJECTIVE: To study the effect of extract of Radix Tetrastigma hemsleyani on the proliferation and apoptosis of human lung carcinoma H1299 cells, and to explore its mechanisms. METHODS H1299 cells were treated with the extract of Radix Tetrastigma hemsleyani in different concentrations at different time points. Its inhibition on H1299 cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. The morphology of the H1299 cell was observed by inverted microscope. Changes of apoptosis were observed by Hoechst33258 methods. The apoptosis rate was detected by flow cytometry. Expression changes of apoptosis-related proteins pro-caspase-3, pro-caspase-9, cle-caspase-3, cle-caspase-9, and poly-ADP-ribose polymerase (PARP) were detected by Western blot. RESULTS: Compared with the control group, the inhibition rate of H1299 cells increased after acted by 0.5, 1, 5, and 10 mg/mL extract of Radix Tetrastigma hemsleyani (P < 0.05, P < 0.01). The longer the acting time, the higher the inhibition rate (P < 0.01). Under inverted microscope, typical morphological changes could be seen and the number of H1299 cells was reduced. Under fluorescence microscope, dark stained nucleus and formed apoptotic body could be observed. Results of flow cytometry showed that the apoptosis rate was obviously dose-effect correlated with the concentration of extract of Radix Tetrastigma hemsleyani. Results of Western blot indicated that compared with the control. group, the protein expression of pro-caspase-3, pro-caspase-9, and PARP were down-regulated and that of cle-caspase-3, cle-caspase-9, and cle-PARP were up-regulated by 5 and 10 mg/mL extract of Radix Tetrastigma hemsleyani (P < 0.05). CONCLUSIONS: Extract of Radix Tetrastigma hemsleyani had obvious effect in inhibiting the proliferation and inducing apoptosis of human lung carcinoma H1299 cells, which might be achieved by activating the expression of caspase protein.

Radix tetrastigma hemsleyani flavone induces apoptosis in human lung carcinoma a549 cells by modulating the MAPK pathway.

Radix Tetrastigma Hemsleyani Flavone (RTHF) is widely used as a traditional herb for its detoxification and anti-inflammation activity. Recently, several studies have shown that RTHF can inhibit growth and induce apoptosis in human cancer cell lines. However, the mechanisms are not completely understood yet. In this study we investigated the potential effects of RTHF on growth and apoptosis in human lung adenocarcinoma A549 cells as well as its mechanisms. A549 cells were treated with RTHF at various concentrations for different times. In vitro the MTT assay showed that RTHF had obvious anti-proliferation effects on A549 cells in a dose- and time- dependent manner. Cell morphological changes observed by inverted microscope and Hoechst33258 methods were compared with apoptotic changes observed by fluorescence microscope. Cell apoptosis inspected by flow cytometry showed significant increase in the treatment group over the control group (P<0.01). Expression of apoptosis related Bax/Bcl-2, caspases and MAPK pathway proteins were detected by Western blotting. The results showed that RTHF up-regulated the Bax/Bcl-2 ratio and cle-caspase3/9, cle-PARP expression in a dose- dependent manner. Expression of p-p38 increased, p-ERK decreased significantly and that of p-JNK was little changed in the RTHF group when compared with the control group. These results suggest that RTHF might exert anti-growth and apoptosis activity against lung cancer A549 cells through activation of caspases and Bcl-2 family proteins and the MAPK pathway, therefore presenting as a promising therapeutic agent for the treatment of lung cancer.

Effect of silicon on the formation of silk fibroin/calcium phosphate composite.

The silk fibroin/calcium phosphate composites were prepared by adding the different amount of Na(2)SiO(3) to assess the effect of silicon on the HA (hydroxyapatite) formation in the composites. FTIR and XRD results suggested that the inorganic phase was constituted mainly by the amorphous DCPD (dicalcium phosphate dehydrate), a precursor of HA in the bone mineral, when the composites were prepared at the final Na(2)SiO(3) concentration lower than 0.008%. Otherwise, HA was formed as the predominant one in the as-prepared composite, accompanied with a conformational transition in the organic phase of silk fibroin protein from silk I (alpha-helix and/or polyglycine II (3(1)-helix) conformations) to silk II (antiparallel beta-sheet conformation). SEM images showed the different morphologies with the samples, i.e., sheet-like crystals in the composites prepared at a low Na(2)SiO(3) concentration and rod-like bundles in other composites. The rod-like bundles were connected together to form the porous network, due to the fact that the HA crystals grew with the aggregation of silk fibroin, and further accreted onto the silk fibroin fibrils. TG curves indicated that the composites prepared with a certain amount of additional SiO (3) (2-) had the higher thermal stability because of its high molecular orientation and crystallinity, and high water-holding capacity due to the porous microstructure.

[Study on treatment of iron-deficiency anemia by shengxuening].

OBJECTIVE: To observe the therapeutic effect of shengxuening (SXN) in treating iron-deficiency anemia (IDA) and to explore its molecular mechanism on iron metabolism balance regulation. METHODS: Patients with IDA were randomly divided into the treated group and the control group, 50 in each group. They were treated with SXN (0.1 g, three times per day) and ferrous gluconate (0.1 g, three times per day) respectively, for 30 days. Levels of serum iron (Fe), total iron binding capacity (TIBC), transferrin saturation (TS), serum ferritin (SF), transferrin (Tf), soluble transferrin receptor (sTfR) and blood routine test, as well as scoring of TCM qi-blood deficiency Syndrome were conducted before and after treatment. RESULTS: The total effective rate in the treated group reached 92%, it was shown that SXN could improve the iron metabolism, increase levels of Fe, TS, SF and reduce levels of TIBC, Tf, sTfR, it has obvious effect in promoting erythrocyte generation and could promote formation of leucocytes and platelets. The total effective rate in the control group was 32%, which was significantly lower than that in the treated group (P < 0.01). CONCLUSION: The effect of SXN in treating IDA and qi-blood deficiency Syndrome is evident, it could improve the iron metabolism, increase levels of Fe, TS, SF and lower levels of TIBC, Tf, sTfR.