[Chaihu Guizhi decoction produces antidepressant-like effects via sirt1-p53 signaling pathway].

OBJECTIVE: To investigate the mechanism of the antidepressant-like effects of Chaihu Guizhi decoction (CGD). OBJECTIVE: Chaihu Guizhi decoction at the daily dose of 17 g/kg and solvent vehicle were administered by gavage in 12 and 14 male C57BL/6J mice for 7 consecutive days, respectively. Forced swimming test (FST), elevated plus maze (EPM) test, open field test (OFT) and novelty-suppressed feeding test (NSF) were performed to assess the depression- and anxiety-like behaviors and motor ability of the mice. We further used chronic social defeat stress (CSDS) and social interaction test to evaluate the antidepressant-like effects of CGD in comparison with the solvent vehicle. Western blotting and RT-qPCR were performed to detect the expressions of sirt1, p53, acetylated p53, and the neuron plasticity-related genes including synapsin I (Syn1), Rab4B, SNAP25 and tubulin beta4b in the hippocampus of the mice. OBJECTIVE: In FST, the immobility time of CGDtreated mice was decreased significantly (P < 0.05); no significant differences were found in the performances in EPM, NSF and OFT tests between the two groups. In social interaction test, the mouse models of CSDS treated with CGD showed significantly increased time in the interaction zone (P < 0.05). Compared with those in the vehicle group, the CGD-treated mouse models exhibited significantly increased protein level of SIRT1 and decreased p53 acetylation (P < 0.05) with up-regulated synapsin I mRNA expression in the hippocampus (P < 0.05); no significant difference were found in Rab (P=0.813), SNAP (P=0.820), or Tubb mRNA expressions (P=0.864) between the two groups. OBJECTIVE: CGD produces antidepressant-like effects in mice possibly through the sirt1-p53 signaling pathway and synaptic plasticity.

Fractional Deletion of Compound Kushen Injection Indicates Cytokine Signaling Pathways are Critical for its Perturbation of the Cell Cycle.

We used computational and experimental biology approaches to identify candidate mechanisms of action of aTraditional Chinese Medicine, Compound Kushen Injection (CKI), in a breast cancer cell line (MDA-MB-231). Because CKI is a complex mixture of plant secondary metabolites, we used a high-performance liquid chromatography (HPLC) fractionation and reconstitution approach to define chemical fractions required for CKI to induce apoptosis. The initial fractionation separated major from minor compounds, and it showed that major compounds accounted for little of the activity of CKI. Furthermore, removal of no single major compound altered the effect of CKI on cell viability and apoptosis. However, simultaneous removal of two major compounds identified oxymatrine and oxysophocarpine as critical with respect to CKI activity. Transcriptome analysis was used to correlate compound removal with gene expression and phenotype data. Many compounds in CKI are required to trigger apoptosis but significant modulation of its activity is conferred by a small number of compounds. In conclusion, CKI may be typical of many plant based extracts that contain many compounds in that no single compound is responsible for all of the bioactivity of the mixture and that many compounds interact in a complex fashion to influence a network containing many targets.

Ligustilide, a major bioactive component of Angelica sinensis, promotes bone formation via the GPR30/EGFR pathway.

Angelica sinensis (Oliv.) Diels is a widely-used traditional Chinese herbal medicine in treating osteoporosis. Ligustilide (LIG) is the main component of A. sinensis and is considered to be the most effective biologically active ingredient in this plant. LIG has been found to have multiple pharmacological activities, such as anti-atherosclerosis, neuroprotection, anticancer, anti-inflammatory and analgesic. However, little is known regarding its anti-osteoporotic effects. The aims of this study were to investigate any protective effect of LIG on bone formation. The results showed that LIG significantly ameliorated inhibition of bone formation in zebrafish caused by prednisolone. LIG promoted osteoblast differentiation, including that of the pre-osteoblastic cell line MC3T3-E1 and bone marrow mesenchymal stem cells. LIG greatly improved the viability of MC3T3-E1 cells exposed to H2O2, attenuated H2O2-induced apoptosis and increased the expression of Bcl-2. Furthermore, LIG treatment lead to marked activation of phosphorylated EGFR and ERK1/2. These effects could be obviously inhibited by blocking GPR30 signaling with the specific inhibitor G15. Collectively, the results reveal that GPR30 is a positive switch for LIG to increase bone formation via regulation of EGFR, and these results provide evidence for the potential of LIG to treat osteoporosis.

Discovery of Allosteric, Potent, Subtype Selective, and Peripherally Restricted TrkA Kinase Inhibitors.

Tropomyosin receptor kinases (TrkA, TrkB, TrkC) are activated by hormones of the neurotrophin family: nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin 3 (NT3), and neurotrophin 4 (NT4). Moreover, the NGF antibody tanezumab has provided clinical proof of concept for inhibition of the TrkA kinase pathway in pain leading to significant interest in the development of small molecule inhibitors of TrkA. However, achieving TrkA subtype selectivity over TrkB and TrkC via a Type I and Type II inhibitor binding mode has proven challenging and Type III or Type IV allosteric inhibitors may present a more promising selectivity design approach. Furthermore, TrkA inhibitors with minimal brain availability are required to deliver an appropriate safety profile. Herein, we describe the discovery of a highly potent, subtype selective, peripherally restricted, efficacious, and well-tolerated series of allosteric TrkA inhibitors that culminated in the delivery of candidate quality compound 23.

Cloning, characterization, and expression analysis of LGP2 cDNA from goose, Anser cygnoides.

Laboratory of genetics and physiology 2 ( LGP2: ) is a homologue of the retinoic acid inducible gene-I and melanoma differentiation associated gene 5 that lacks the caspase activation and recruitment domain required for signaling. It plays a pivotal role in host immune response. In this study, we cloned and characterized the full-length open reading frame ( ORF: ) sequence of LGP2 in the Qingyuan goose (Anser cygnoides) and evaluated the mRNA expression of this gene post infection with an H5N1 highly pathogenic avian influenza virus ( HPAIV: ). The full-length goose LGP2 ORF (2,028 bp) encoded a polypeptide of 675 amino acids. The deduced amino acid sequence contained 5 main overlapping structural domains-2 DEAD/DEAH box helicase domains, one conserved restriction domain of bacterial type III restriction enzyme, one helicase superfamily C-terminal domain and one C-terminal regulatory domain. Quantitative real-time PCR analysis indicated that goose LGP2 was constitutively expressed in all 19 investigated tissues, but the expression level was different among them. It was high expressed in the trachea, jejunum, bursa, kidney and heart, but low in the glandular stomach, lung, liver, spleen, crop and muscular stomach. A significant increase in the transcription of LGP2 was detected in the brain, spleen and lungs of geese post infection with H5N1 HPAIV versus uninfected tissues. These findings indicated that goose LGP2 was an important receptor that is involved in the host antiviral innate immune defense to H5N1 HPAIV in geese.

Computational identification and characterization of conserved miRNAs and their target genes in beet (Beta vulgaris).

Highly conserved endogenous non-coding microRNAs (miRNAs) play important roles in plants and animals by silencing genes via destruction or blocking of translation of homologous mRNA. Sugar beet, Beta vulgaris, is one of the most important sugar crops in China, with properties that include wide adaptability and strong tolerance to salinity and impoverished soils. Seedlings of B. vulgaris can grow in soils containing up to 0.6% salt; it is important to understand the molecular mechanisms of salt tolerance to enrich genetic resources for breeding salt-tolerant sugar beets. Here, we report 13 mature miRNAs from 12 families, predicted using an in silico approach from 29,857 expressed sequence tags and 279,223 genome survey sequences. The psRNATarget server predicted 25 target genes for the 13 miRNAs. Most of the target genes appeared to encode transcription factors or were involved in metabolism, signal transduction, stress response, growth, and development. These results improve our understanding of the molecular mechanisms of miRNA in beet and may aid in the development of novel and precise techniques for understanding post-transcriptional gene-silencing mechanisms in response to stress tolerance.

1,25(OH)2D3 downregulates the Toll-like receptor 4-mediated inflammatory pathway and ameliorates liver injury in diabetic rats.

BACKGROUND: Fatty acid deposition in the liver can activate a number of pro-inflammatory signaling pathways such as the Toll-like receptor 4 (TLR4) pathway, which may be important in the pathogenesis of nonalcoholic steatohepatitis. 1,25(OH)2D3 downregulates the expression of TLR4 and may represent a novel treatment strategy for reducing hepatocyte injury. Therefore, in this study, we investigated the protective effects of 1,25(OH)2D3 on diabetic liver injury in vivo. METHODS: Streptozotocin (STZ)-induced diabetic rats were randomly divided into five groups and treated with low-dose 1,25(OH)2D3 (0.025 µg/kg/day), medium-dose 1,25(OH)2D3 (0.15 µg/kg/day), high-dose 1,25(OH)2D3 (0.3 µg/kg/day), insulin (protamine zinc insulin 16 U/kg/day, subcutaneous injection), or no intervention (the control group). Sixteen weeks later, the rats were killed, and blood samples were obtained to test lipid profiles and hepatic function. The infiltration of inflammatory cells, the level of fibrosis, and the expression levels of TLR4, nuclear factor-kappa B (NF-κB), and tumor necrosis factor-α (TNF-α) in the liver were analyzed. The hepatocytes were treated with vehicle control, LPS (100 ng), high fat [DMEM + FFA (0.1 mM: palmitic acid, oleic acid, 1:2)], LPS + high fat, vehicle + 1,25(OH)2D3 (10(-7) M), LPS + 1,25(OH)2D3, high fat + 1,25(OH)2D3, or LPS + high fat + 1,25(OH)2D3. RNA and protein were extracted to detect the expression of TLR4 and downstream inflammatory factors such as NF-ΚB, TNF-α, and IL-6. Groups of data were compared by single factor variance analysis. RESULTS: High-dose 1,25(OH)2D3 administration for 16 weeks downregulated the expression of TLR4, NF-κB, and TNF-α in the liver tissue of diabetic rats and attenuated hepatic inflammation and fibrosis, as shown by immunohistochemical staining, hematoxylin and eosin staining, Masson's trichrome staining, reverse transcription polymerase chain reaction (RT-PCR), and western blotting. In vitro, hepatocytes treated with high fat or LPS exhibited significantly increased expression of TLR4, NF-κB, and downstream inflammatory factors (P < 0.05). Intervention with 1,25(OH)2D3 decreased the expression of TLR4, NF-κB, and inflammatory factors (P < 0.05). CONCLUSIONS: 1,25(OH)2D3 exhibited protective effects against diabetes-related liver injury, possibly through downregulation of components of the TLR4 signaling pathway.

Sequential therapy for non-thalamus supratentorial hypertensive intracerebral hemorrhages.

OBJECTIVE: We sought to assess the effectiveness of sequential therapy for non-thalamus supratentorial hypertensive intracerebral hemorrhage (NTS-HICH). PATIENTS AND METHODS: We retrospectively analyzed clinical data of 110 patients with HICH. The patients were admitted 72 hours after disease onset, and 43 patients received sequential therapy. The length of hospital stay, treatment costs, incidence of pulmonary infections, mortality rates and Modified Rankin Score (mRS) 1 and 3 months after NTS-HICH were compared between patients who received sequential or non-sequential therapies. RESULTS: The length of hospital stay, treatment costs, and 1-month mortality rates were not significantly different between both groups. However, mortality rates at 3 months, incidence of pulmonary infection, and mRS at both 1 and 3 months were significantly better in patients who received sequential therapy. CONCLUSIONS: Sequential therapy significantly improves the prognosis for patients with NTS-HICH.

Correlation between oxidative stress and L-type calcium channel expression in the ventricular myocardia of selenium-deficient mice.

OBJECTIVE: Expression of the Cacna1c (calcium channel, voltage-dependent, L-type, α1C subunit) gene was studied to investigate the relationship between oxidative stress and L-type calcium channels in the myocardium of selenium-deficient mice. METHODS: Selenium levels in liver and heart tissue samples from mice fed normal or selenium-deficient diets were evaluated by fluorometry. In the same mice, glutathione peroxidase (GPx) and Cacna1c gene expression were analysed, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were measured, oxidoreductase gene expression profiles were analysed (by DNA microarray), and myocardial structural changes were studied. RESULTS: In selenium-deficient versus control mice, GPx expression and SOD activity were decreased, and Cacna1c expression and MDA concentration were increased. Selenoprotein oxidoreductase and nonselenoprotein oxidoreductase gene expression differed significantly between selenium-deficient and control mice. In selenium-deficient mice, myocardial fibres were separated by loose collagenous tissue; electron microscopy showed shortened sarcomeres, dilated sarcoplasmic reticulum, scattered myofibril lysis and increased mitochondria with amorphous matrix densities. CONCLUSION: L-type calcium channels were involved in selenium deficiency-induced cardiomyocyte injury, which was positively related to oxidative stress.

Hypothalamic suppressor-of-cytokine-signalling 3 mRNA is elevated and pro-opiomelanocortin mRNA is reduced during pregnancy in Brandt's voles (Lasiopodomys brandtii ).

Leptin acts within the hypothalamus to diminish food intake. In Brandt's voles (Lasiopodomys brandtii), both circulating leptin levels and food intake are elevated during pregnancy, suggesting an ineffectiveness of leptin to reduce food intake. Diminished hypothalamic leptin receptors and impaired leptin signal transduction are characteristic of central leptin resistance. The present study aimed to determine whether these characteristic modulations of leptin sensitivity occurred in pregnant Brandt's voles. The mRNA expression of the long form of the leptin receptor (Ob-Rb), suppressor-of-cytokine-signalling 3 (SOCS3), neuropeptide Y (NPY), agouti-related protein (AgRP), pro-opiomelanocortin (POMC) and cocaine- and amphetamine-regulated transcript (CART) in the hypothalamus were examined on dioestrous, day 5, day 10 and day 18 of pregnancy. Compared to controls, there was no significant change in hypothalamic Ob-Rb mRNA during the pregnancy. SOCS3 mRNA was increased significantly by 68% on day 10% and 93% on day 18 of pregnancy compared to controls. Despite elevated leptin levels, POMC mRNA was decreased significantly by 60% on day 18 of pregnancy, whereas no differences were found in the mRNA expression of NPY, AgRP and CART in pregnant voles compared to controls. The elevation of SOCS3 mRNA together with disrupted leptin regulation of neuropeptides in the hypothalamus suggests that leptin resistance may develop in pregnant Brandt's voles.

First Report of Anthracnose of Lycium barbarum Caused by Colletotrichum acutatum in China.

Barbary wolfberry (Lycium barbarum, Solanaceae) is an important Chinese traditional medicine that is widely planted in northwestern China (6.7 × 104 ha under cultivation, including Ningxia Hui Autonomous Region). After a recent, large increase in the planting area and density, anthracnose has become more damaging. In China, Colletotrichum gloeosporioides was assumed to be the sole causal agent of anthracnose on L. chinense (wolfberry) (3), whereas in Korea, C. dematium was reported to cause anthracnose on wolfberry (4). During the summer and autumn of 2007, 29 barbary wolfberry fruit samples were collected from three orchards in Zhongning County, Ningxia Hui Autonomous Region. Conidia were 8.5 to 16.5 × 2.5 to 4 µm and fusiform or pointed at one or both ends. Slow-growing colonies on potato dextrose agar were white to orange or pink; sclerotia and setae were absent. The morphological traits were identical to those of C. acutatum and clearly distinct from those of C. gloeosporioides (conidia cylindrical with both ends rounded, gray colony color) or C. dematium (conidia falcate, sclerotia and setae abundant) (2-4). Koch's postulates were performed to verify that the isolates were capable of causing anthracnose on wolfberry. Six wolfberry fruits were surface sterilized with 70% alcohol, allowed to dry 1 min, then wounded with a sterile needle, and dipped in 6 ml of spore suspension (1 × 105 conidia/ml). Anthracnose symptoms were observed on inoculated fruit after 3 days, whereas control fruits inoculated with sterile water did not develop symptoms. The pathogenicity test was performed three times; in each trial, fungi reisolated from symptomatic tissue were morphologically identical to those that had been used as inoculum. Amplification of the internal transcribed spacer (ITS) region of rDNA with primers ITS1 and ITS4 resulted in bands of approximately 600 bp. The sequences of both isolates were compared with sequences deposited in the GenBank database and demonstrated 99% similarity to C. acutatum isolate DQ286123. PCR amplification of the ITS region was also carried out using species-specific primer CaInt2 in conjunction with the universal primer ITS4 (1). A DNA fragment of approximately 500 bp was amplified from all isolates, whereas no amplification products were obtained from reference cultures of C. gloeosporioides and C. dematium. To our knowledge, this is the first report of C. acutatum causing anthracnose on L. barbarum. References: (1) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (2) B. C. Sutton. Page 523 in: The Coelomycetes. Commonwealth Mycological Institute, Kew, Surrey, England, 1980. (3) X. M. Wang and J. Y. Li. Acta Mycol. Sinica 6:211, 1987. (4) S. H. Yu. Korean J. Plant Pathol. 2:31, 1986.

Alleviation of osteoarthritis by calycosin-7-O-beta-D-glucopyranoside (CG) isolated from Astragali radix (AR) in rabbit osteoarthritis (OA) model.

OBJECTIVE: This work was undertaken to assess the protective effect of an isoflavonoid, calycosin-7-O-beta-D-glucopyranoside (CG), isolated from Astragali radix (AR) on the pathogenesis of osteoarthritis (OA)-like lesion in a rabbit model. METHODS: Nine rabbits underwent an anterior cruciate ligament and menisectomy transection (ACLMT) of the rear knee joints to induce OA-like lesion. They were randomly divided into three groups (n=6/group): a negative control group treated with 200 microl of 0.5% (v/v) dimethyl sulfoxide (DMSO), a positive control group treated with 200 microl of 100 microM piroxicam, and a test group treated with 100 microg/500 microl of CG, where the test agents were administered by injection once a week for 4 weeks starting from the third week. Rabbits were then sacrificed to observe the progression of OA-like lesion. The synovial fluid was analyzed for the amounts of total proteins, glycosaminoglycans (GAG) and prostaglandin E(2) (PGE(2)). In addition, histopathologic analyses were performed on the OA-like articular cartilage with or without therapeutic treatments. RESULTS: The total synovial fluid volume (P<0.05) was most strikingly reduced by the treatment with CG. Moreover, the CG treatment also significantly alleviated the OA-induced accumulation of prostaglandin (PG) (P<0.001) and total proteins (P<0.001) in the synovial fluid. The histopathologic analyses revealed that the CG treatment reduced the severity of the OA-like structural damages in the cartilage. However, the level of PGE(2), a pathologic inflammatory molecule, was not diminished by CG or piroxicam. CONCLUSION: These results indicate that the isoflavonoid CG isolated from AR significantly alleviated the pathologic changes in the OA-like rabbit knee joints. This suggests that CG from AR could be a promising treatment for the therapy of OA.

MicroRT-small animal conformal irradiator.

A novel small animal conformal radiation therapy system has been designed and prototyped: MicroRT. The microRT system integrates multimodality imaging, radiation treatment planning, and conformal radiation therapy that utilizes a clinical 192Ir isotope high dose rate source as the radiation source (teletherapy). A multiparameter dose calculation algorithm based on Monte Carlo dose distribution simulations is used to efficiently and accurately calculate doses for treatment planning purposes. A series of precisely machined tungsten collimators mounted onto a cylindrical collimator assembly is used to provide the radiation beam portals. The current design allows a source-to-target distance range of 1-8 cm at four beam angles: 0 degrees (beam oriented down), 90 degrees, 180 degrees, and 270 degrees. The animal is anesthetized and placed in an immobilization device with built-in fiducial markers and scanned using a computed tomography, magnetic resonance, or positron emission tomography scanner prior to irradiation. Treatment plans using up to four beam orientations are created utilizing a custom treatment planning system-microRTP. A three-axis computer-controlled stage that supports and accurately positions the animals is programmed to place the animal relative to the radiation beams according to the microRTP plan. The microRT system positioning accuracy was found to be submillimeter. The radiation source is guided through one of four catheter channels and placed in line with the tungsten collimators to deliver the conformal radiation treatment. The microRT hardware specifications, the accuracy of the treatment planning and positioning systems, and some typical procedures for radiobiological experiments that can be performed with the microRT device are presented.

Synthesis and biological activities of diquaternary dipiperazinium salts containing dithiocarboxyl groups.

A series of diquaternary dipiperazinium salts containing dithiocarboxyl groups 6a-f and 9 were synthesized and evaluated for their analgesic and sedative activities. The result showed that the presence of two quaternary ammonium cations and the distance between them are very important for the activities of the salts. Compound 6b exhibited the best activities (at dose 2 mg/kg, analgesic, 57%; sedative, 59%) among compounds 6a-f. Compound 9 not only showed the most potent analgesic (85.4%, dose 1 mg/kg) and sedative (93.1%, dose 1 mg/kg) activities, but also exhibited anticancer activity against KB (68.7%, dose 10 microM).

[Effects of Rholida on the free radical metabolism and serum creatine kinase after exercise at plateau].

Objective. To study the effect of Rholida on free radical metabolism and serum creatine kinase CK) after exercise at plateau. Method. After staying at high altitude (4100 m) for 20 d, 40 healthy young men were divided into 4 groups randomly (Rholida, Acetazolamide, Xi' s capsule and control, 10 men each group). And their SOD, MDA, GSH-Px CK, and CK-MB were determined respectively. Before, after taking drugs and after finishing the 5 min-stair-exercise. Result. Before taking drugs and after exercise, MAD GSH-Px, CK, CK-MB, increased as compared with quiet state (P<0.05, P<0.01), but SOD showed no significant chang (P>0.05). After taking drugs for 6 d, those who took Rholida, Acetazolamide and Xi's capsule, their MAD, GSH-Px CK, CK-MB increased after exercise as compared with quiet state (P<0.05). In Rholida group SOD increased and had significant change (P<0.05); but there was no significant change in Acetazolamide, Xi' s capsule group, SOD increased, MDA decreased (P<0.05), CK, CK-MB had no significant change (P>0.05), GSH-Px increased in Xi's group (P<0.05), but not in Acetazolamide group (P>0.05). SOD, GSH-Px increased, MDA, CK-MB decreased in Rholida group after taking drugs and the changes were significant (P<0.01). In Acetazolamide and Xi's capsule group, GSH-Px increased significantly, MDA, CK, CK-MB decreased significantly (P<0.05), but SOD didn't (P>0.05). Conclusion. Rholida, Acetazolamide, Xi's capsule could regulate the disorder of free radical metabolism at plateau and Rholida had advantage over the others.

[Study on the protective effect of xinkang oral liquid on the isoproterenol-induced ischemic myocardial damage in rats].

OBJECTIVE: To study the protective effect of Xinkang oral liquid on the ischemic myocardial damage. METHODS: Ischemic myocardial damage model was obtained by i.p. 85 mg/kg isoproterenol. The level of superoxide dismutase (SOD), malondialdehyde (MDA), creatine kinase (CK), lactic dehydrogenase (LDH) in the serum and endothelin (ET) in the plasma were detected. RESULTS: The level of ET, MDA, CK, LDH of Xinkang oral liquid group was much lower than that of model group (P < 0.01). The SOD activity was higher than that of model group (P < 0.01). The level of Xinkang oral liquid group was lower than that of Shuxin oral liquid group. The SOD activity of Xinkang oral liquid group was much higher than that of Shuxin oral liquid group (P < 0.01). CONCLUSION: Xinkang oral liquid can prevent the myocardium from ischemic damage.

Two diastereomeric saponins with cytotoxic activity from Albizia julibrissin.

Two diastereomeric saponins, julibrosides J1 (1) and J9 (2), both of which show cytotoxic activity, were obtained from the stem bark of Albizia julibrissin Durazz. On the basis of chemical and spectral evidence [L.B. Ma et al., Carbohydr. Res., 281 (1996) 35-46], the structure of 1 was revised as 3-O-[beta-D-xylopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-(1-->6) -beta-D-glucopyranosyl]-21-O-[(6S)-2-trans-2-hydroxymethyl-6-methyl-6-O- [4-O-((6R)-2-trans-2,6-dimethyl-6-O-(beta-D-quinovopyranosyl)-2,7- octadienoyl)-beta-D-quinovopyranosyl]-2,7-octadienoyl] acacic acid-28-O-beta-D-glucopyranosyl-(1-->3)-[alpha-L-arabinofuranosyl-(1-->4 )]-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl ester. The diastereoisomer 2 of 1 was identified as 3-O-[beta-D-xylopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-(1-->6) -beta-D-glucopyranosyl]-21-O-[(6S)-2-trans-2-hydroxymethyl-6-methyl-6-O- [4-O-((6S)-2-trans-2,6-dimethyl-6-O-(beta-D-quinovopyranosyl)-2,7- octadienoyl)-beta-D-quinovopyranosyl]-2,7-octadienoyl] acacic acid-28-O-beta-D-glucopyranosyl-(1-->3)-[alpha-L-arabinofuranosyl-(1-->4 )]-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl ester. Saponin 2 is a new saponin named julibroside J9. Both julibrosides J1 and J9 show good inhibitory action against the KB cancer cell line in vitro.

Inhibitory effect of antisense epidermal growth factor receptor RNA on the proliferation of rat C6 glioma cells in vitro and in vivo.

OBJECT: The goal of this study was to evaluate the effect of antisense epidermal growth factor receptor (EGFR) RNA on the growth of rat glioma cells in vitro and in vivo and to determine the feasibility of targeting the EGFR gene for gene therapy in gliomas. METHODS: Antisense EGFR complementary (c)DNA was transfected into C6 glioma cells by using lipofectamine. In vitro studies, Southern and Northern blot analyses, in situ hybridization, and immunohistochemical staining were designed to examine the integration and expression of antisense EGFR constructs. The 3'(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and the average number of argyrophilic nuclear organizer regions (Ag-NORs) were used to evaluate cell proliferation, whereas the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) method and microscopy were used to observe cell apoptosis. As part of the in vivo studies, parental C6 cells and C6 cells transfected with EGFR antisense cDNA were implanted stereotactically into the right caudate nucleus of Wistar rats (C6-injected animals and transfected C6-injected animals). Rats with well-established cerebral C6 glioma foci were treated intratumorally with either antisense EGFR cDNA or empty-vector DNA by using lipofectamine (treated-C6 and control treated group). The general behavior and survival of the rats, findings on magnetic resonance images of their brains, histopathological changes, proliferation activity, and apoptosis of the cerebral gliomas in each group of rats were examined. Exogenous antisense EGFR cDNA was integrated into the genome of C6 cells and expressed. In clones with a high expression of the antisense construct, there was a dramatic decrease in endogenous EGFR messenger RNA and protein levels, reduced proliferation activity, and induction of apoptosis in vitro. The mean survival time of rats injected with C6 cells was 17.3 days. The mean survival time of rats injected with C6 cells followed by treatment with empty vector in lipofectamine was 15.4 days. Survival time was significantly prolonged in 100% of the rats injected with antisense-transfected C6 cells and in two thirds of the rats injected with C6 cells followed by antisense EGFR cDNA. Magnetic resonance imaging revealed distinct cerebral tumor foci in C6-injected rats and in control rats of the treated group, but none were found in the rats injected with transfected C6 cells. Furthermore, tumor foci disappeared completely in C6-injected rats treated with antisense EGFR cDNA. The cerebral gliomas of the rats treated by injection of antisense EGFR RNA were characterized by reduced proliferation activity and the induction of apoptosis. CONCLUSIONS: The results of this study indicate that EGFR plays an important role in the genesis of malignant gliomas. It may, therefore, be an effective target of antisense gene therapy in patients with gliomas.

[Chinese drugs for invigorating spleen to remove dampness, activating blood circulation to eliminate turbid in retarding progression of chronic renal failure].

OBJECTIVE: To investigate the effect of Chinese drugs (CD) for invigorating Spleen to remove dampness, activating blood circulation to eliminate Turbid in retarding progression of chronic renal failure (CRF). METHODS: Thirty-nine patients with CRF were divided into two groups randomly, 18 patients in the control group were treated with low protein diet and controlling blood pressure and 21 patients in the CD group were treated similarly with that of the control group but with additional CD. Levels of serum creatinine (SCr), blood urea nitrogen (BUN), blood albumin (Alb), cholesterol (CH), triglyceride (TG), high-density lipoprotein (HDL) and hemoglobin (Hb) were checked every 2 months, and the rate of progression of CRF was estimated by slope of the creatinine reciprocal (1/SCr) with time (month). RESULTS: Levels of SCr and BUN in the CD group were significantly lower and HDL markedly higher than those in the control group, P < 0.05. Mean slopes of the creatinine reciprocal with time in the two groups was significantly different, P < 0.01. CONCLUSION: Additional CD treatment based upon the protein diet and controlling blood pressure could retard the progression process of CRF evidently.

Gynostemma pentaphyllum: identification of major sapogenins and differentiation from Panax species.

Four main dammarane-type aglycones of gypenosides, extracted from the aerial parts of Gynostemma pentaphyllum were identified by gas chromatography-mass spectrometry. By detecting these aglycones as well as the aglycones of ginsenosides, a difference in sapogenin composition between Gynostemma pentaphyllum and Panax species was observed, which can be used in the differentiation of these plant drugs.