Investigation of insecticidal activity of two Rhododendron species on stored-product insects.

This study was designed to investigate the insecticidal activity of the essential oils (EOs) and extracts from Rhododendron rufum and Rhododendron przewalskii. The EOs were extracted from the leaves of R. Rufum and R. przewalskii by hydro-distillation and their chemical components were analyzed by gas chromatography-mass spectrometry (GC-MS). The repellency, contact toxicity and antifeedant activity of the EOs and extracts were evaluated against Sitophilus oryzae and Tribolium castaneum along with those of their main components. A total of nine compounds were identified from the EO of R. Rufum, and the most abundant component was myristicin (79.72%). The EO of R. Rufum exhibited repellent activities at different levels and its main compound myristicin showed contact toxicity and repellent effects against S. oryzae and T. castaneum. Meanwhile, by bioassay-guided fractionation, four compounds with strong antifeedant activities against T. castaneum, 24-methylenecycloartanyl-2'E, 4'Z-tetradecadienoate (1), methyl thyrsiflorin B acetate (2), friedelin (3) and Excoecarin R1 methyl ester (4) were separated and identified from the ethanol extract of R. przewalskii for the first time. Considering the significant anti-insect activities, the EOs and extracts of R. Rufum and R. przewalskii might be used in integrated pest strategies, establishing a good perspective for the comprehensive use of natural plant resources of Rhododendron genus.

Implantation of computed tomography-guided high-dose-rate 192Ir brachytherapy in oldest old patients with advanced non-small cell lung cancer: A case report and literature review.

BACKGROUND: With the increasing aging and the popularization of medical diagnosis, the growing number of oldest old with lung cancer needs to be focused on. Several medical and physiological challenges often accompanying the oldest old cancer patients make the choice of the optimal treatment daunting. The current research suggests that people who get adequate treatment can benefit, but it is worth discussing which treatment will benefit them more. High-dose-rate (HDR) 192Ir brachytherapy deserves attention in this context owing to its association with less trauma and reduced complications. CASE PRESENTATION: An 86-years-old woman with a right glandular lung carcinoma presented with progressive lesions 11 months after chemotherapy. Because of her old age and poor performance status (eastern cooperative oncology group performance status 3), she received HDR 192Ir brachytherapy for her right lung lesion without any common complications, such as pneumothorax and hemorrhage. She continued on 0.25 g oral gefitinib each day after received brachytherapy treatment. The right lung lesion keeps a partial response until 18 months later now. CONCLUSION: HDR 192Ir brachytherapy can potentially be used as a safe and effective choice for the oldest old with advanced non-small cell lung cancer. It can especially benefit cancer patients with concurrent chemotherapy or targeted therapy.

[Textual research on classical prescriptions in Mongolian medicine].

Mongolians have a long history of using prescriptions, which can be classified into four stages as follows: the germination and experience accumulation stage before the 13 th century, the theoretical formation stage from the 13 th to 16 th century, the rapid development stage from the 17 th to 20 th century, and the leaping development stage from the mid-20 th century to the present. The prescriptions from the ancient classical or representative medical books have always been used by Mongolian physicians for generations, and they are still in use due to the definite curative effects. In 2008, the Notice on Issuing the Supplementary Provisions to the Registration and Management of Traditional Chinese Medicine(TCM) described that China has attached more importance to the excavation and development of classical prescriptions. As stipulated in the Law of the People's Republic of China on Traditional Chinese Medicine, the classical prescriptions should be those available in ancient TCM classics and still in wide use, with exact curative effects, distinct features, and obvious advantages. This paper expounded the historical formation and development of classical prescriptions in Mongo-lian medicine, introduced the five most influential ancient medical books revealing the formation and development of these classic prescriptions, and traced the origin of such classical prescriptions as Wenguanmu Siwei Decoction, Shouzhangshen Bawei Decoction, Jianghuang Siwei Decoction and summarized the origin, development history and characteristics of classical prescriptions in Mongolian medicine, aiming to provide a reference for their further research and development.

Age-related changes of normal prostate: evaluation by MR diffusion tensor imaging.

In this study, fifty healthy normal volunteers were divided into 3 groups according to age: group A (15-30 years, n=14), group B (31-50 years, n=24), group C (>51 years, n=12). The FA and ADC values in PZ and CZ were measured, and difference between the PZ and CZ were assessed. The results indicated that no significant difference were found in the FA and ADC values between the left and right of PZ (P>0.05), but significant differences were observed in the FA and ADC values between PZ and CZ within each group (P<0.05). The FA values of PZ in three groups were 0.227±0.052, 0.202±0.055, and 0.145±0.034, respectively. The ADC values were found to be 1.439±0.160×10(-3), 1.652±0.256×10(-3), and 2.001±0.266×10(-3) mm(2)/s, accordingly. The FA and ADC values in PZ were significantly (P<0.05) different between groups. The FA values of CZ in different groups were found to be 0.291±0.083, 0.287±0.045, and 0.257±0.059, respectively; while the corresponding ADC values were 1.374±0.171×10(-3), 1.382±0.178×10(-3), and 1.415±0.136×10(-3) mm(2)/s, respectively. The FA and ADC values in CZ were not statistically (P>0.05) different between groups. Pearson correlation analysis results showedthat the FA values in PZ havenegative correlation with age (r=-0.498, P<0.05), while the ADC values exhibited a positive correlation with age (r=0.682, P<0.05). No correlations between the changes of FA and ADC values and age were noted in CZ. In conclusion, the FA and ADC values in the normal prostatic PZ were age-dependent. FA decreases and ADC increases with age. In contrast, the FA and ADC values in the normal prostatic CZ were not significantly age-related.

[Study on effects and mechanism of traditional Mongolian medicine wuweifengshi capsule on adjuvant arthritis in rats].

OBJECTIVE: To study the effects and immunoregulation mechanism of the traditional Mongolian medicine Wuweifengshi capsule on adjuvant arthritis (AA). METHOD: Wister rats were divided into several groups: normal group, AA model group, Wuweifengshi capsule groups (with low, moderate, high dose of 0.2, 0.4, 0.8 g x kg(-1) x d(-1) respectively), and Zhonglun-5 group (original dose of 1.68 g x kg(-1) x d(-1)). The edema degree, the level of IL-1beta, TNF-alpha, PGE2, NO and MDA and the activity of SOD in serum were detected. Through cell culture, the effects of the medicine on AA rat's splenic cell's multiplication capacity were studied. The influence of celiac macrophage cell culture fluid of AA rats' on C57BL/6J mice thymic cell multiplication capacity under the medicine was evaluated. RESULT: Wuweifengshi capsule showed an inhibiting function on the level of IL-1beta, TNF-alpha, PGE2, NO and increased the activity of SOD in serum, but showed no significant influence on MDA. It also inhibited the AA rat's splenic cell's multiplication capacity and the influence of celiac macrophage cell culture fluid of AA rat's on C57BL/6J mice thymic cell multiplication capacity. CONCLUSION: The anti-AA effect of Wuweifengshi capsule is possibly due to its inhibition of relevant cytokines and its adjustment of corresponding enzyme's activity and immunization organ's cell multiplication capacity.

Cotton AnnGh3 encoding an annexin protein is preferentially expressed in fibers and promotes initiation and elongation of leaf trichomes in transgenic Arabidopsis.

The annexins are a multifamily of calcium-regulated phospholipid-binding proteins. To investigate the roles of annexins in fiber development, four genes encoding putative annexin proteins were isolated from cotton (Gossypium hirsutum) and designated AnnGh3, AnnGh4, AnnGh5, and AnnGh6. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) results indicated that AnnGh3, AnnGh4, and AnnGh5 were preferentially expressed in fibers, while the transcripts of AnnGh6 were predominantly accumulated in roots. During fiber development, the transcripts of AnnGh3/4/5 genes were mainly accumulated in rapidly elongating fibers. With fiber cells further developed, their expression activity was dramatically declined to a relatively low level. In situ hybridization results indicated that AnnGh3 and AnnGh5 were expressed in initiating fiber cells (0-2 DPA). Additionally, their expression in fibers was also regulated by phytohormones and [Ca(2+)]. Subcellular localization analysis discovered that AnnGh3 protein was localized in the cytoplasm. Overexpression of AnnGh3 in Arabidopsis resulted in a significant increase in trichome density and length on leaves of the transgenic plants, suggesting that AnnGh3 may be involved in fiber cell initiation and elongation of cotton.

Cotton plasma membrane intrinsic protein 2s (PIP2s) selectively interact to regulate their water channel activities and are required for fibre development.

Aquaporins are thought to be associated with water transport and play important roles in cotton (Gossypium hirsutum) fibre elongation. Among aquaporins, plasma membrane intrinsic proteins (PIPs) constitute a plasma-membrane-specific subfamily and are further subdivided into PIP1 and PIP2 groups. In this study, four fibre-preferential GhPIP2 genes were functionally characterized. The selective interactions among GhPIP2s and their interaction proteins were studied in detail to elucidate the molecular mechanism of cotton fibre development. GhPIP2;3 interacted with GhPIP2;4 and GhPIP2;6, but GhPIP2;6 did not interact with GhPIP2;4. Coexpression of GhPIP2;3/2;4 or GhPIP2;3/2;6 resulted in a positive cooperative effect which increased the permeability coefficient of oocytes, while GhPIP2;4/2;6 did not. GhBCP2 (a blue copper-binding protein) inhibited GhPIP2;6 water channel activity through their interaction. Overexpression of GhPIP2 genes in yeast induced longitudinal growth of the host cells. By contrast, knockdown of expression of GhPIP2 genes in cotton by RNA interference markedly hindered fibre elongation. In conclusion, GhPIP2 proteins are the primary aquaporin isoforms in fibres. They selectively form hetero-oligomers in order to regulate their activities to meet the requirements for rapid fibre elongation.

A cotton gene encoding MYB-like transcription factor is specifically expressed in pollen and is involved in regulation of late anther/pollen development.

In flowering plants, pollen development is a highly programmed process, in which a lot of genes are involved. In this study, a gene, designated as GhMYB24, encoding R2R3-MYB-like protein was isolated from cotton. GhMYB24 protein is localized in the cell nucleus and acts as a transcriptional activator. Northern blot analysis revealed that GhMYB24 transcripts were predominantly detected in anthers. It was further found that strong expression of GhMYB24 was mainly detected in pollen and was regulated during anther development by in situ hybridization. Overexpression of GhMYB24 in Arabidopsis caused flower malformation, shorter filaments, non-dehiscent anthers and fewer viable pollen grains. Further analysis revealed that the septum and stomium cells of anthers were not broken, and fewer fibrous bands were found in the endothecium cells in transgenic plants. A complementation test demonstrated that GhMYB24 was able to recover partially the male fertility of the myb21 myb24 double mutant. Expression levels of the genes involved in the phenylpropanoid biosynthetic pathway and reactive oxygen species homeostasis were altered in GhMYB24-overexpressing transgenic plants. Furthermore, the genes involved in jasmonate biosynthesis and its signaling pathway were up-regulated in the transgenic plants. Yeast two-hybrid assay indicated that GhMYB24 interacted with GhJAZ1/2 in cells. Taking the data together, our results suggest that GhMYB24 may play an important role in normal anther/pollen development.

A cotton LIM domain-containing protein (GhWLIM5) is involved in bundling actin filaments.

LIM-domain proteins play important roles in cellular processes in eukaryotes. In this study, a LIM protein gene, GhWLIM5, was identified in cotton. Quantitative RT-PCR analysis showed that GhWLIM5 was expressed widely in different cotton tissues and had a peak in expression during fiber elongation. GFP fluorescence assay revealed that cotton cells expressing GhWLIM5:eGFP fusion gene displayed a network distribution of eGFP fluorescence, suggesting that GhWLIM5 protein is mainly localized to the cell cytoskeleton. When GhWLIM5:eGFP transformed cells were stained with rhodamine-phalloidin there was consistent overlap in eGFP and rhodamine-palloidin signals, demonstrating that GhWLIM5 protein is colocalized with the F-actin cytoskeleton. In addition, high-speed cosedimentation assay verified that GhWLIM5 directly bound actin filaments, while low cosedimentation assay and microscopic observation indicated that GhWLIM5 bundled F-actin in vitro. Increasing amounts of GhWLIM5 protein were able to protect F-actin from depolymerization in vitro in the presence of Lat B (an F-actin depolymerizer). Our results contribute to a better understanding of the biochemical role of GhWLIM5 in modulating the dynamic F-actin network in cotton.

Two new flavonoid glycosides from Artemisia frigida Willd.

An investigation of the n-BuOH-soluble fraction from the aerial parts of Artemisia frigida has led to the isolation of two new flavonoid glycosides, named friginoside A and friginoside B. Their structures were characterized as 5,7-dihydroxy-3',4',5'-trimethoxy flavone 7-O-ß-d-glucuronide (1) and 5,7-dihydroxy-3',4',5'-trimethoxyflavone 7-O-ß-d-glucuronyl-(1 â†’ 2)O-ß-d-glucuronide (2) on the basis of 1D and 2D NMR spectral analysis.

Three cotton genes preferentially expressed in flower tissues encode actin-depolymerizing factors which are involved in F-actin dynamics in cells.

To investigate whether the high expression levels of actin-depolymerizing factor genes are related to pollen development, three GhADF genes (cDNAs) were isolated and characterized in cotton. Among them, GhADF6 and GhADF8 were preferentially expressed in petals, whereas GhADF7 displayed the highest level of expression in anthers, revealing its anther specificity. The GhADF7 transcripts in anthers reached its peak value at flowering, suggesting that its expression is developmentally-regulated in anthers. The GhADF7 gene including the promoter region was isolated from the cotton genome. To demonstrate the specificity of the GhADF7 promoter, the 5'-flanking region, including the promoter and 5'-untranslated region, was fused with the GUS gene. Histochemical assays demonstrated that the GhADF7:GUS gene was specifically expressed in pollen grains. When pollen grains germinated, very strong GUS staining was detected in the elongating pollen tube. Furthermore, overexpression of GhADF7 gene in Arabidopsis thaliana reduced the viable pollen grains and, consequently, transgenic plants were partially male-sterile. Overexpression of GhADF7 in fission yeast (Schizosaccharomyces pombe) altered the balance of actin depolymerization and polymerization, leading to the defective cytokinesis and multinucleate formation in the cells. Given all the above results together, it is proposed that the GhADF7 gene may play an important role in pollen development and germination.

The GhACS1 gene encodes an acyl-CoA synthetase which is essential for normal microsporogenesis in early anther development of cotton.

Microsporogenesis, associated with the functional expression of many genes, is a highly programmed and regulated process in flowering plants. To elucidate the roles of genes during anther development, two anther-specific cDNAs (designated GhACS1 and GhACS2) encoding acyl-CoA synthetases (ACSs) were isolated from a cotton (Gossypium hirsutum) flower cDNA library. Subsequently, the corresponding GhACS1 gene was isolated from a cotton genomic DNA library. Real-time quantitative RT-PCR and northern blot analyses revealed that GhACS1 transcripts were predominantly accumulated in the developing anthers of cotton. The specificity of GhACS1 expression in primary sporogenous cells (PSCs), pollen mother cells (PMCs), microspores, and tapetal cells was demonstrated by in situ hybridization as well as histochemical assay of GUS expression controlled under the GhACS1 promoter. High levels of GhACS1 activity are crucial for fatty acid metabolism in PSCs, PMCs, microspores and particularly tapetal cells. Reduction of ACS enzymatic activity by suppressing GhACS1 expression severely affected the tapetal cells and consequently blocked normal microsporogenesis in early anther development. Aberrant and defective microspores were generated in the transgenic anthers. As a result, the transgenic plants failed to produce functional pollen grains and were male-sterile, suggesting that the GhACS1 gene is required for normal microsporogenesis in early anther development of cotton.

Characterization of 19 novel cotton FLA genes and their expression profiling in fiber development and in response to phytohormones and salt stress.

Fasciclin-like arabinogalactan proteins (FLAs), a subclass of arabinogalactan proteins (AGPs), are usually involved in cell development in plants. To investigate the expression profiling as well as the role of FLA genes in fiber development, 19 GhFLA genes (cDNAs) were isolated from cotton (Gossypium hirsutum). Among them, 15 are predicted to be glycosylphosphatidylinositol anchored to the plasma membranes. The isolated cotton FLAs could be divided into four groups. Real-time quantitative reverse transcriptase polymerase chain reaction results indicated that the GhFLA genes are differentially expressed in cotton tissues. Three genes (GhFLA1/2/4) were specifically or predominantly expressed in 10 days post-anthesis fibers, and the transcripts of the other four genes (GhFLA6/14/15/18) were accumulated at relatively high levels in cotton fibers. Furthermore, expressions of the GhFLA genes are regulated in fiber development and in response to phytohormones and NaCl. The identification of cotton FLAs will facilitate the study of their roles in cotton fiber development and cell wall biogenesis.

[Characterization and expression analysis of two cotton genes encoding putative UDP-Glycosyltransferases].

UDP-Glycosyltransferases (UGT) are a large family of enzymes, which catalyze the transfer of a sugar from an activated sugar donor to an acceptor molecule. Both in plant and in mammalian, they are important in maintenance of cellular homeostasis. In this study, two genes (designated GhUGT1 and GhUGT2, respectively) encoding putative UGT were isolated from cotton fiber cDNA library. The deduced proteins contain the signature sequences of plant UGTs in the C-terminal region. The GhUGT1 gene encodes a polypeptide of 457 amino acids, and displays homology at amino acid levels with the known glucosyltransferase genes. Sequence analysis revealed that the GhUGT2 merely encodes a small protein, as there is a nucleotide substitution that results in formation of a stop codon in its open-reading frame. Real-time RT-PCR analysis revealed that the expression of GhUGT1 is higher in the fast growth tissues, such as in fibers and roots. GhUGT2 has also higher expression in roots, but with lower expression levels in fibers and other tissues. The result also showed that the expression of GhUGT1 is higher than GhUGT2. Further study showed that GhUGT1 and GhUGT2 expressions are regulated under osmotic stress, suggesting they may be involved in plants responding to osmotic stress.

Molecular characterization of cotton GhTUA9 gene specifically expressed in fibre and involved in cell elongation.

The microtubule cytoskeleton may play an important role in the polarized growth of fibre cells that are single-cell trichomes on the surface of cotton ovules. To investigate whether the high expression levels of alpha-tubulin genes are correlated with fibre elongation, nine GhTUA genes (cDNAs) encoding alpha-tubulins with 449-451 amino acid residues were isolated and characterized in cotton. The GhTUA genes share high sequence homology at the nucleotide level (62-93% identity) in the coding region and at the amino acid level (89-99% identity), and can be classified into two subgroups. Real-time quantitative RT-PCR analysis revealed that seven out of the nine GhTUA genes are predominantly expressed in developing fibres. Among them, GhTUA9 displays the highest level of expression, revealing its fibre specificity. The GhTUA9 transcripts in fibres reached its peak value between 5-10 DPA, and dramatically declined to undetectable levels as the ovule matured further, suggesting that its expression is developmentally-regulated in fibres. The GhTUA9 gene including the promoter region was isolated from the cotton genome. To demonstrate the specificity of the GhTUA9 promoter, the 5'-flanking region, including the promoter and 5'-untranslated region, was fused with the GUS gene. Histochemical assays demonstrated that the GhTUA9:GUS gene was specifically expressed in elongating fibres. Overexpression of GhTUA9 in fission yeast (Schizosaccharomyces pombe) promoted atypical longitudinal growth of the host cells by 1.4-1.7-fold, indicating that the GhTUA9 gene is involved in cell elongation. Given all the above results, it is proposed that the GhTUA9 gene may play an important role in fibre elongation.

Molecular characterization and expression analysis of nine cotton GhEF1A genes encoding translation elongation factor 1A.

The translation elongation factor 1A, eEF1A, plays an important role in protein synthesis, catalyzing the binding of aminoacyl-tRNA to the A-site of the ribosome by a GTP-dependent mechanism. To investigate the role of eEF1A for protein synthesis in cotton fiber development, nine different cDNA clones encoding eukaryotic translation elongation factor 1A were isolated from cotton (Gossypium hirsutum) fiber cDNA libraries. The isolated genes (cDNAs) were designated cotton elongation factor 1A gene GhEF1A1, GhEF1A2, GhEF1A3, GhEF1A4, GhEF1A5, GhEF1A6, GhEF1A7, GhEF1A8, GhEF1A9, respectively. They share high sequence homology at nucleotide level (71-99% identity) in the coding region and at amino acid level (96-99% identity) among each other. Phylogenetic analysis demonstrated that the nine GhEF1A genes can be divided into 5-6 subfamilies, indicating the divergence occurred in structures of the genes as well as the deduced proteins during evolution. Real-time quantitative RT-PCR analysis revealed that GhEF1A genes are differentially expressed in different tissues/organs. Of the nine GhEF1A genes, five are expressed at relatively high levels in young fibers. Further analysis indicated that expressions of the GhEF1As in fiber are highly developmental-regulated, suggesting that protein biosynthesis is very active at the early fiber elongation.

[Cloning of GhAQP1 gene and its specific expression during Ovule development in cotton].

Plant aquaporins, belonging to the MIP superfamily, are a series of transmembrane proteins that facilitate water transport through cell membranes. In this study, a cDNA clone encoding the PIP1-like protein was isolated from cotton (Gossypium hirsutum) cDNA libraries, and designated as GhAQP1 (Fig.1). We also isolated the GhAQP1 gene from cotton genome by PCR. The gene is 2,096 bp in length, including an open reading frame (ORF) and 5'-/3'-untranslated regions (UTR). It contains two introns in its ORF. The first intron is inserted between codons 209 and 210 in the fifth transmembrane helix, and another is located between codons 256 and 257 in the sixth transmembrane helix of GhAQP1, respectively (Figs.2 and 3). Northern blot analysis showed that GhAQP1 gene is expressed specifically in 6-15 DPA ovule, and reaches a peak in 9 DPA ovule (Figs.4 and 5), suggesting that its expression is ovule-specific and developmentally regulated in cotton.

The cotton ACTIN1 gene is functionally expressed in fibers and participates in fiber elongation.

Single-celled cotton fiber (Gossypium hirsutum) provides a unique experimental system to study cell elongation. To investigate the role of the actin cytoskeleton during fiber development, 15 G. hirsutum ACTIN (GhACT) cDNA clones were characterized. RNA gel blot and real-time RT-PCR analysis revealed that GhACT genes are differentially expressed in different tissues and can be classified into four groups. One group, represented by GhACT1, is expressed predominantly in fiber cells and was studied in detail. A 0.8-kb GhACT1 promoter sufficient to confirm its fiber-specific expression was identified. RNA interference of GhACT1 caused significant reduction of its mRNA and protein levels and disrupted the actin cytoskeleton network in fibers. No defined actin network was observed in these fibers and, consequently, fiber elongation was inhibited. Our results suggested that GhACT1 plays an important role in fiber elongation but not fiber initiation.