FAP deficiency enhances thermogenesis and attenuates metabolic inflammation in diet-induced obesity.

OBJECTIVE: Fibroblast activation protein α (FAP) is expressed in normal adipose tissue and related to some pleiotropic metabolic regulators. However, the exact role and mechanism of FAP in obesity and related metabolic disorders are not well understood. METHODS: FAP knockout mice were fed a normal diet or a high-fat diet (HFD) for 12 weeks. FAP knockout mice or wild-type mice treated with an FAP inhibitor were subjected to cold stress for 5 days. RESULTS: FAP deficiency protected mice against HFD-induced obesity and obesity-associated metabolic dysfunction, including glucose intolerance, insulin resistance, hyperglycemia, hyperinsulinemia, and hyperlipidemia. Notably, FAP deficiency largely reversed obesity-induced adipose tissue macrophage accumulation and M1-M2 imbalance in white adipose tissue (WAT). Moreover, energy expenditure was significantly higher in FAP-deficient mice fed an HFD. Both FAP deficiency and inhibition increased cold tolerance through enhancing WAT beiging. CONCLUSIONS: This study demonstrated that FAP deficiency protects mice against diet-induced obesity and related metabolic dysfunction. Furthermore, the protective effects are probably mediated via the promotion of WAT beiging and suppression of inflammation.

PIK3CA mutation affects the proliferation of colorectal cancer cells through the PI3K-MEK/PDK1-GPT2 pathway.

The phosphatidylinositol­3­kinase catalytic subunit α (PIK3CA) gene is mutated in numerous human cancers. This mutation promotes the proliferation of tumor cells; however, the underlying mechanism is still not clear. In the present study, it was revealed that the PIK3CA mutation in colorectal cancer (CRC) HCT116 (MUT) rendered the cells more dependent on glutamine by regulating the glutamic­pyruvate transaminase 2 (GPT2). The dependence of glutamine increased the proliferation of cells in a normal environment and resistance to a suboptimal environment. Further study revealed that the mutated PIK3CA could regulate GPT2 expression not only through signal transduction molecule 3­phosphoinositide­dependent kinase (PDK1) but also through mitogen­activated protein kinase (MEK) molecules. In HCT116 cells, MEK inhibitor treatment could reduce the expression of GPT2 signaling molecules, thereby inhibiting the proliferation of CRC cells. A new signal transduction pathway, the PI3K/MEK/GPT2 pathway was identified. Based on these findings, MEK and PDK1 inhibitors were combined to inhibit the aforementioned pathway. It was revealed that the combined application of MEK and PDK1 inhibitors could promisingly inhibit the proliferation of MUT compared with the application of PI3K inhibitors, PDK1 inhibitors, or MEK inhibitors alone. In vivo, MEK inhibitors alone and combined inhibitors had stronger tumor­suppressing effects. There was no significant difference between the PDK1­inhibitor group and normal group in vivo. Thus, these results indicated that mutated PI3K affected GPT2 mediated by the MEK/PDK1 dual pathway, and that the PI3K/MEK/GPT2 pathway was more important in vivo. Inhibiting MEK and PDK1 concurrently could effectively inhibit the proliferation of CRC cells. Targeting the MEK and PDK1 signaling pathway may provide a novel strategy for the treatment of PIK3CA­mutated CRC.

A comparative morphological and transcriptomic study on autotetraploid Stevia rebaudiana (bertoni) and its diploid.

Stevia rebaudiana is an important medical plant for producing steviol glycosides (SGs) or stevioside. Autotetraploids (4x = 44) show an increasing level of morphology, physiology and tolerances comparing to diploids (2x = 22). However, little information regarded on the comparative transcriptome analysis between diploid and autotetraploid S. rebaudiana was found. In this study, synthetic autotetraploid was induced and morphological features were confirmed. A comprehensive transcriptome of stevia leaf, stem and root from the diploids and autotetraploids was constructed based on RNA-seq, yielded 1,000,892,422 raw reads and subsequently assembled into 251,455 transcripts, corresponded to 146,130 genes. Pairwise comparisons of the six leaf libraries between the diploids and autotetraploids revealed 4114 differentially expression genes (DEGs), in which 2105 (51.17%) were up-regulated in autotetraploids and associated with SGs biosynthesis, plant growth and secondary metabolism. Moreover, weighted gene co-expression network analysis showed co-expressed genes of fifteen genes of SG biosynthesis pathway were enriched in photosynthesis, flavonoid and secondary metabolic process, plant growth and morphogenesis. A hundred of DEGs related to plant resistance were identified by interviewing PlantPReS database. This study has highlighted molecular changes related to SGs metabolism of polyploidy, and advanced our understanding in plant resistance responsible for phenotypic change of autotetraploids.

De novo transcriptome sequencing and analysis of genes related to salt stress response in Glehnia littoralis.

Soil salinity is one of the major environmental stresses affecting plant growth, development, and reproduction. Salt stress also affects the accumulation of some secondary metabolites in plants. Glehnia littoralis is an endangered medicinal halophyte that grows in coastal habitats. Peeled and dried Glehnia littoralis roots, named Radix Glehniae, have been used traditionally as a Chinese herbal medicine. Although Glehnia littoralis has great ecological and commercial value, salt-related mechanisms in Glehnia littoralis remain largely unknown. In this study, we analysed the transcriptome of Glehnia littoralis in response to salt stress by RNA-sequencing to identify potential salt tolerance gene networks. After de novo assembly, we obtained 105,875 unigenes, of which 75,559 were annotated in public databases. We identified 10,335 differentially expressed genes (DEGs; false discovery rate <0.05 and |log2 fold-change| ≥ 1) between NaCl treatment (GL2) and control (GL1), with 5,018 upregulated and 5,317 downregulated DEGs. To further this investigation, we performed Gene Ontology (GO) analysis and the Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis. DEGs involved in secondary metabolite biosynthetic pathways, plant signal transduction pathways, and transcription factors in response to salt stress were analysed. In addition, we tested the gene expression of 15 unigenes by quantitative real-time PCR (qRT-PCR) to confirm the RNA-sequencing results. Our findings represent a large-scale assessment of the Glehnia littoralis gene resource, and provide useful information for exploring its molecular mechanisms of salt tolerance. Moreover, genes enriched in metabolic pathways could be used to investigate potential biosynthetic pathways of active compounds by Glehnia littoralis.

In vitro and in vivo aphrodisiac properties of the seed extract from Allium tuberosum on corpus cavernosum smooth muscle relaxation and sexual behavior parameters in male Wistar rats.

BACKGROUND: Allium tuberosum is a well-known spice as well as a herb in traditional Chinese medicine, used for increasing libido and treating erectile dysfunction. However, not many studies have been done to evaluate the sexual enhancing properties of A. tuberosum. The aim of this study was to evaluate the aphrodisiac and vasorelaxant properties of A. tuberosum on corpus cavernosum smooth muscle (CCSM) as well as checking the effect on enhancing male rat sexual behavior, libido, potency as well as its spermatogenic properties. METHOD: The seeds were powdered and sequentially extracted with hexane, ethyl acetate and butanol. Male Wistar rats were administered with graded doses of the n-BuOH extracts (ATB) of A. tuberosum (50, 100, 200 and 400 mg/kg) and Viagra was used as the positive control drug. The extract/drug was administered by gastric probe once daily for 45 days and the sexual behavior was analyzed by exposing the male rats to female rats in the estrus period. RESULTS: ATB relaxed corpus cavernosum smooth muscle (68.9%) at a concentration of 200 µg/ml. The results obtained from the animal studies indicated that ATB significantly increased mount frequency (MF), intromission frequency (IF), ejaculation frequency (EF), ejaculation latency (EL) and markedly reduced post ejaculatory interval (PEI), mount latency (ML), and intromission latency (IL). Furthermore, a remarkable increase in the test for potency was observed as witnessed by marked increase in erections, quick flips, long flips and total reflex. In addition, ATB significantly improved the sperm viability and count as well as increased the concentrations of testosterone, follicle stimulating hormone (FSH), and phosphatases in the treated animals. CONCLUSION: Thus our results suggest that A. tuberosum could stimulate sexual arousal and enhance sexual execution in male rats, thus providing valuable experimental evidence that A. tuberosum possesses sexual enhancing properties.

Allium tuberosum: Antidiabetic and hepatoprotective activities.

Allium tuberosum (AT) is traditionally used for treating nocturnal emissions, abdominal pain, diarrhea, sexual dysfunction and asthma. This study aimed at investigating the antidiabetic and hepatoprotective activities of the butyl alcohol fraction from the methanolic extract of A. tuberosum. For the antidiabetic activity, rats were induced with diabetes by intraperitoneal injection of 150mg/kg alloxan and treated for 30days with AT extract (100, 200 and 400mg/kg). Animals were sacrificed after the study and the fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), HDL, malondialdehyde (MDA) catalase, superoxide dismutase and glutathione levels were determined. The hepatoprotective assay, mice were pretreated for seven days with AT (100, 200 and 400mg/kg) and silymarin (100mg/kg or). Thereafter 10ml/kg of 2% v/v CCl4 was administered intraperitoneally on the 7th day to induce acute liver injury. Blood and liver samples were obtained and serum enzymes ALT, AST, ALP, SOD, GSH, CAT, MDA and pro-inflammatory mediators were assessed. AT significantly decrease FBG, serum TG, TC, MDA levels and significant increased HDL, SOD, GSH and CAT activities in the diabetic rats. In addition, AT significantly inhibited MDA, IL-1b, IL-6 and TNF-α levels and prevented the depletion of the antioxidant enzymes GSH, SOD and CAT activities in CCl4 induced liver damage. Furthermore, AT markedly reduced AST, ALT and ALP levels in the CCl4 treated mice groups. In conclusion, the antidiabetic and hepatoprotective effect of AT may be associated with its antioxidant and its ability to inhibit the pro-inflammatory mediators.

[Effect of exogenous Ca2+ on protective infection of Pinellia ternata and accumulation of major components under high temperature stress].

OBJECTIVE: To study the effect of exogenous Ca2+ on protective infection of Pinellia ternata and accumulation of major components under high temperature stress. METHOD: The soilless cultivation experiment was applied, stress resistance index of P. ternata leaves, statistics the rate of lodge P. ternata,the content of oxalate in different places in the plant, the content of total alkaloids, total organic acids and glucosine in P. ternata tubers were measured based on different concentrations of exogenous Ca2+. RESULT: The test results showed that, at lower concentrations of Ca2+ treatments, the rate of lodge P. ternata was higher than that of the others. With Ca2+ concentration increasing, activities of SOD and POD initially increased and then decreased, however, proline level tended to be down then up. Soluble oxalic acid content was lower than the content of unhandled treatment in P. ternata leaves and tubers; with Ca2+ concentration increasing, soluble oxalic acidl content and yield showed a tendency of decrease after increase in the leaves and tubers. Compared with other treatments, spraying 400 mg x L(-1) Ca2+ significantly enhanced the accumulation of total alkaloid and guanosine in P. ternata tubers. At Lower concentrations of Ca2+, the content of total free organic acid was higher in the tuber. CONCLUSION: With the treatment of Ca2+ the capacity of heat resistance was improved in P. ternata plants, the rate of lodge P. ternata was postponed, growing period was extended and corresponding production has increased by spraying exogenous Ca2+.

[Study on salt stress tolerance of Chrysanthemum morifolium 'Hangbaiju' and 'Huangju' and F1 seedlings].

OBJECTIVE: To study the salt stress tolerance of Hongxinju, Huangju and F1 seedlings from orthogonal and reciprocal cross under different salt treatments. Grope for transmissibility of salt tolerance between parents and F1 seedlings, and relativity between flavone, chlorogenic acid contents and salt tolerance. METHOD: The materials were put in 5 different concentrations of Hoagland nutrient solution (0, 40, 80, 120, 160 mmol x L(-1)) containing NaCl, keeping grads while raising the consistency of NaCl day by day. The injured leaf area per plant, proline, betaine, MDA, flavones and chlorogenic acid contents were measured and analyzed after treatment. RESULT: As NaCl concentration was below 120 mmol x L(-1), the salt tolerance of Hongxinju was higher than that of Huangju, the salt tolerance of Hongxinju x Huangju higher than that of parents, the salt tolerance of Huangju x Hongxinju was at the level of parents. As NaCl concentration between 120 to 160 mmol x L(-1), the salt tolerance of Huangju was higher than that of Hongxinju, the salt tolerance of Huangju x Hongxinju higher than that of parents and the salt tolerance of Hongxinju x Huangju was at the level of parents. CONCLUSION: Salt tolerance of F1 is more influenced by female parent, relativity showed between flavonoids, chlorogenic acid contents and salt tolerance.

[Selection of the microorganism for dioscin hydrolyze].

OBJECTIVE: To select the microorganism which can hydrolyze dioscin to diosgenin. METHODS: The microorganism were selected from the surface of rhizome, rhizosphere soil, the inside of the leaves and rhizome of Dioscorea zingiberensis C. H. Wright. Diosgenin was identified by thin-layer chromatography and HPLC. RESULTS: The microorganism which could hydrolyze dioscin from the experiment were identified as Aspergillus sp and Alternaria sp. Characteristics of enzymes production and fermentation technology of Aspergillus No. 1 were also studied primarily. CONCLUSION: The Aspergillus strain No. 1 can secret enzyme to hydrolyze dioscin into diosgenin effectively.