Specific roles of Os4BGlu10, Os6BGlu24, and Os9BGlu33 in seed germination, root elongation, and drought tolerance in rice.

MAIN CONCLUSION: Morphological, physiological, and gene expression analyses showed that Os4BGlu10, Os6BGlu24, and Os9BGlu33 played specific roles in seed germination, root elongation, and drought tolerance of rice, with various relations with indole-3-acetic acid (IAA) and abscisic acid (ABA) signaling. ß-Glucosidases (BGlus) belong to glycoside hydrolase family 1 and have many functions in plants. In this study, we investigated the function of three BGlus in seed germination, drought tolerance, and root elongation using the loss-of-function mutants bglu10, bglu24, and bglu33. These mutants germinated slightly later under normal conditions and had significantly longer roots than the wild type. In the presence of ABA, bglu10 and bglu24 exhibited a higher germination inhibition percentage, whereas bglu33 had a lower germination inhibition percentage, compared to the wild type. All of the mutants exhibited less drought tolerance, with the survival rates significantly lower than that of the wild type, which was also confirmed by a decrease in relative leaf water content and Fv/Fm ratio after drought treatment. The root length of bglu10 did not respond to IAA, whereas that of bglu24 responded to a high (0.25 µM) concentration of IAA, and that of bglu33 to a low (0.05 µM) concentration of IAA. The root length of bglu10 and bglu24 did not respond to ABA, whereas that of bglu33 increased significantly in response to a high (0.05 µM) concentration of ABA. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that expression of Os4BGlu10 was up-regulated by polyethylene glycol (PEG), whereas that of Os6BGlu24 was up-regulated by 0.25 µM IAA, and Os9BGlu33 was up-regulated by PEG, IAA, and ABA. Taken together, we demonstrate that Os4BGlu10, Os6BGlu24, and Os9BGlu33 play specific roles in seed germination, root elongation, and drought tolerance with various relation with IAA and ABA signaling.

Traditional Chinese medicine Astragalus reverses predominance of Th2 cytokines and their up-stream transcript factors in lung cancer patients.

Th2 cytokine is predominant in tumor patients and was found to be associated with tumor progression. Reversing of Th2 dominant status is thought to be a promising strategy. In the present study, peripheral blood mononuclear cells (PBMNC) of 37 lung cancer patients and 19 healthy subjects were prepared and used for examination of cytokine secretion and gene expression. The positive percentage of mRNA transcripts of Th1 cytokines (8.1% for IFNgamma and 13.5% for IL-2) in patients' PBMNC were lower than those of Th2 cytokines (70.3% for IL-4, 64.9% for IL-6 and 83.8% for IL-10). The gene expression capacity (measured as relative intensity to ratio of beta-actin) of patients for Th1 cytokines was low, but constitutively relatively high for Th2 cytokines. Both positive percentage and relative intensity were lower in transcript factor for Th1 cytokine, T-bet (40.5% and 0.139, respectively) than those for Th2 cytokine, GATA3 (89.2% and 0.364, respectively). Traditional Chinese medicine, Astragalus (AG) was observed to reverse Th2 status of lung cancer. AG enhanced culture supernatant and gene expression levels of Th1 cytokine (IFNgamma and IL-2) and its transcript factor (T-bet), and reduced those of Th2 cytokines in cultured PBMNC of lung cancer patients. These results demonstrated that traditional Chinese medicine AG might reverse the Th2 predominant status in lung cancer patients, which is a probable alternative therapeutic regime in future.

Type two cytokines predominance of human lung cancer and its reverse by traditional Chinese medicine TTMP.

Type 2 cytokines are usually predominant in tumor patients and associated with tumor progression. To explore whether reversing of type 2 predominance could be a promising strategy in tumor immunotherapy, PBMC of 35 lung cancer patients and 19 healthy subjects were prepared and subjected to be examined for cytokine secretion and gene expression. Tetra-Methylpyrazine (TTMP), extracted from a traditional Chinese medicinal herb which has been used in clinic to reverse the Th2 status of cancer patients in China, was added to PBMC culture. Determined by RT-PCR, the positive percentages of mRNA expression of type 1 cytokines (8.6% for IFN-gamma and 11.4% for IL-2) were lower than those of type 2 cytokines (71.4% for IL-4, 60% for IL-6 and 80% for IL-10) in patients' PBMCs. The potential of gene expressing (measured as relative intensity to the ratio of beta-actin) in the patients for type 1 cytokines was also in a low level (0.111 for IFN-gamma, 0.119 for IL-2) in comparison with a relative high level for type 2 cytokines (0.319 for IL-4, 0.303 for IL-6 and 0.377 for IL-10). Meanwhile, both positive percentage and relative intensity of gene expression were lower for a type 1 cytokine-related transcription factor T-bet (31.4% and 0.142, respectively) than those for type 2 cytokine-related GATA3 (85.7% and 0.378, respectively). The blood serum levels of IFN-gamma and IL-2 in the patients were slightly lower but not significantly when compared with healthy control. In contrast, the levels IL-4 and IL-6 in patients were significantly higher than those in healthy subjects by ELISA analysis. TTMP could enhance supernatant concentration and gene expression levels of IFN-gamma, IL-2 and T-bet, but reduce those of type 2 cytokines. These results demonstrate that the lung cancer patients had a predominant expression of type 2 cytokines and TTMP could reverse the type 2 dominant status, which might offer an alternative therapeutic regime for lung cancer patients.