Canagliflozin alleviates pulmonary hypertension by activating PPARγ and inhibiting its S225 phosphorylation.

Pulmonary hypertension (PH) is a progressive fatal disease with no cure. Canagliflozin (CANA), a novel medication for diabetes, has been found to have remarkable cardiovascular benefits. However, few studies have addressed the effect and pharmacological mechanism of CANA in the treatment of PH. Therefore, our study aimed to investigate the effect and pharmacological mechanism of CANA in treating PH. First, CANA suppressed increased pulmonary artery pressure, right ventricular hypertrophy, and vascular remodeling in both mouse and rat PH models. Network pharmacology, transcriptomics, and biological results suggested that CANA could ameliorate PH by suppressing excessive oxidative stress and pulmonary artery smooth muscle cell proliferation partially through the activation of PPARγ. Further studies demonstrated that CANA inhibited phosphorylation of PPARγ at Ser225 (a novel serine phosphorylation site in PPARγ), thereby promoting the nuclear translocation of PPARγ and increasing its ability to resist oxidative stress and proliferation. Taken together, our study not only highlighted the potential pharmacological effect of CANA on PH but also revealed that CANA-induced inhibition of PPARγ Ser225 phosphorylation increases its capacity to counteract oxidative stress and inhibits proliferation. These findings may stimulate further research and encourage future clinical trials exploring the therapeutic potential of CANA in PH treatment.

Effect of Shading and Forest Type on Morphological Characteristics and Bioactive Compounds of Fruiting Bodies of Songshan Lingzhi Ganoderma tsugae (Agaricomycetes).

Ganoderma tsugae strain MCCCMAS0053 cultivation on short logs results in varied yield and quality under different growth conditions. Thus, growth conditions need optimization to increase yield and quality. An indoor experiment with three shade treatments (A1, two layers of black sun-shade net; A2, one layer of black sun-shade net plus plastic mulch; A3, one layer of black sun-shade net) and a field experiment in two forest types (pine or mixed pine-oak) were conducted. The results showed that shading and forest type significantly affected light intensities and the growth, size, biomass, and bioactive components of G. tsugae fruiting bodies. In the indoor experiment, the mean dry weight of the fruiting body and the diameter of the pileus in A2 increased by 21.51-44.98% and 13.42-22.26%, respectively, compared with those of A1 and A3. Similarly, the accumulation of the bioactive compounds (polysaccharides, total amino acids, and total essential amino acids) in the pileus and stipe were greater in A2 than in A1 and A3. Furthermore, compared with pure pine forest cultivation, fruiting bodies cultivated in mixed pine-oak forest had greater dry weight, pileus diameter, and more bioactive compounds. In addition, no significant difference was found between the A2 and mixed pine-oak treatments, which had a similar light intensity (from 1116 to 2367 lx). Hence, this suggests that the A2 shade treatment or cultivation in mixed pine-oak forest is beneficial for production of G. tsugae fruiting bodies, and light intensity may play a critical role in this process.

Hydroxy-octadecenoic acids instead of phorbol esters are responsible for the Jatropha curcas kernel cake's toxicity.

The toxic kernel cake of Jatropha curcas (KCakeJ) is an emerging health and environmental concern. Although phorbol esters are widely recognized as the major toxin of KCakeJ, convincing evidence is absent. Here, we show that rather than phorbol esters an isomeric mixture of 11-hydroxy-9E-octadecenoic acid, 12-hydroxy-10E-octadecenoic acid and 12-hydroxy-10Z-octadecenoic acid (hydroxy-octadecenoic acids, molecular formula C18H34O3) is the major toxic component. The toxicities of hydroxy-octadecenoic acids on experimental animals, e.g. acute lethality, causing inflammation, pulmonary hemorrhage and thrombi, allergies, diarrhea and abortion, are consistent with those on human/animals caused by Jatropha seed and/or KCakeJ. The hydroxyl group and the double bond are essential for hydroxy-octadecenoic acids' toxicity. The main pathway of the toxicity mechanism includes down-regulating UCP3 gene expression, promoting ROS production, thus activating CD62P expression (platelet activation) and mast cell degranulation. The identification of the major toxin of KCakeJ lays a foundation for establishing an environmentally friendly Jatropha biofuel industry.

[Digoxin inhibits migration and invasion of human gastric carcinoma MKN45 cells through down-regulation of AEG-1].

This study was designed to investigate the effect of digoxin on migration and invasion of human gastric carcinoma MKN45 cells and its possible mechanism. MKN45 cells were treated with different concentrations of digoxin for 24 h. The shRNA-AEG-1 plasmid was transfected into MKN45 cells via lipofectamine to block the expression of astrocyte elevated gene-1 (AEG-1). Western blot was used to analyze the protein levels of matrix metalloproteinase-9 (MMP-9), E-cadherin and AEG-1. The result showed that digoxin reduced the abilities of migration and invasion (P < 0.05), up-regulated the protein level of E-cadherin (P < 0.05), and down-regulated the protein levels of MMP-9 and AEG-1 (P < 0.05) in MKN45 cells in a dose-dependent manner. Compared with shControl group, shAGE-1 group showed inhibited cellular migration and invasion, higher expression level of E-cadherin, and lower expression levels of MMP-9 and AEG-1. These results suggest that digoxin suppresses the migration and invasion of human gastric carcinoma MKN45 cells in a dose-dependent manner through inhibiting the expression of AEG-1, and then resulting in the up-regulation of the protein expression of E-cadherin and the down-regulation of the protein expression of MMP-9.

[Clinical and Pathological Analysis of 9 Patients with Primary Breast Diffuse Large B-cell Lymphoma].

OBJECTIVE: To explore the clinical features, diagnosis and treatment of primary breast diffuse large B- cell lymphoma (PBDLBCL). METHODS: Clinical records of 9 PBDLBCL patients treated in Department of Hematology of Yijishan Hospital Affiliated to Wannan Medical College from August 2001 to January 2014 were analyzed retrospectively. RESULTS: All of the 9 patients were female, with an average age of 48 years (range 28 to 75), 8 cases had unilateral breast tumors and 1 case had bilateral. According to the Ann Arbor stage standard, 2 cases were of stage IE and 7 were IIE. None of them was concurrent with B symptoms; 6 cases had IPI (International prognostic Index) score 0 and 3 had score 1. 2 cases belonged to germinal center B cells type (GCB) and 7 belonged to non-GCB. Double-Hit lymphomas were presented in 3 cases. Out of 9 cases, 3 cases were diagnosed by using tubular needle biopsy, 5 cases were diagnosed by using resection of breast mass, and 1 case was diagnosed by using modified radical mastectomy. 1 case received radical mastectomy, 1 case received unilateral breast removal, 1 case gave up, 1 case received mass excision with chemotherapy and radiotherapy, 5 cases received mass excision with chemotherapy and 1 case received central prophylaxis. A complete response (CR) was observed in 6 cases after first-line chemotherapy. The median follow-up time was 18 months (range 0.1 to 150), 3 cases relapsed and 5 cases died. CONCLUSION: PBDLBCL mostly occurs in female. The main pathological type is non-GCB coupled with Double-Hit lymphoma. Tubular needle biopsy offers benifit in the diagnosis of PBL, R-CHOP or R-CHOP combined with chemotherapy/radiotherapy produce best outcome among all the treatments. Intrathecal injection of chemotherapy drugs may help to prevent recurrence of PBL central.

Differential gene expression of the key signalling pathway in para-carcinoma, carcinoma and relapse human pancreatic cancer.

Pancreatic cancer (PC) has a high rate of mortality and a poorly understood mechanism of progression. Investigation of the molecular mechanism of PC and exploration of the specific markers for early diagnosis and specific targets of therapy are key points to prevent and treat PC effectively and to improve their prognosis. In our study, expression profiles experiment of para-carcinoma, carcinoma and relapse human PC was performed using Agilent human whole genomic oligonucleotide microarrays with 45 000 probes. Differentially expressed genes related with PC were screened and analysed further by Gene Ontology term analysis and Kyoto encyclopaedia of genes and genomes pathway analysis. Our results showed that there were 3853 differentially expressed genes associated with pancreatic carcinogenesis and relapse. In addition, our study found that PC was related to the Jak-STAT signalling pathway, PPAR signalling pathway and Calcium signalling pathway, indicating their potential roles in pancreatic carcinogenesis and progress.

Synthesis and antimicrobial activity of polyhalobenzonitrile quinazolin-4(3H)-one derivatives.

A novel series of polyhalobenzonitrile quinazolin-4(3H)-one derivatives were synthesized and characterized by NMR, IR, MS, and HRMS spectra. All of the newly prepared compounds were screened for antimicrobial activities against four strains of bacteria (Gram-positive bacterial: Staphylococcus aureus and Bacillus cereus; Gram-negative bacterial: Escherichia coli and Pseudomonas aeruginosa) and one strain of fungi (Candida albicans). Among the synthesized compounds, 5-(dimethylamino)-8-(2,4,5-trichloro-isophthalonitrile) quinazolin-4(3H)-one (7k) exhibited significant activity towards Gram-positive bacterial, Gram-negative bacterial, and the fungi strains. The MIC (0.8-3.3µg/mL) and MBC (2.6-7.8µg/mL) for this compound were close to those of nofloxacin, chlorothalonil, and fluconazole, making it the most potent antimicrobial agents in the series.

Detoxification of Jatropha curcas kernel cake by a novel Streptomyces fimicarius strain.

A huge amount of kernel cake, which contains a variety of toxins including phorbol esters (tumor promoters), is projected to be generated yearly in the near future by the Jatropha biodiesel industry. We showed that the kernel cake strongly inhibited plant seed germination and root growth and was highly toxic to carp fingerlings, even though phorbol esters were undetectable by HPLC. Therefore it must be detoxified before disposal to the environment. A mathematic model was established to estimate the general toxicity of the kernel cake by determining the survival time of carp fingerling. A new strain (Streptomyces fimicarius YUCM 310038) capable of degrading the total toxicity by more than 97% in a 9-day solid state fermentation was screened out from 578 strains including 198 known strains and 380 strains isolated from air and soil. The kernel cake fermented by YUCM 310038 was nontoxic to plants and carp fingerlings and significantly promoted tobacco plant growth, indicating its potential to transform the toxic kernel cake to bio-safe animal feed or organic fertilizer to remove the environmental concern and to reduce the cost of the Jatropha biodiesel industry. Microbial strain profile essential for the kernel cake detoxification was discussed.

Synthesis and antimicrobial activity of polyhalo isophthalonitrile derivatives.

A series of polyhalo isophthalonitrile derivatives (3 and 4) that incorporate a variety of substituents at the 2-, 4-, 5- and/or 6-positions of the isophthalonitrile moieties have been designed and synthesized. These derivatives were evaluated for their antimicrobial activity against Staphylococcus aureus, Bacillus cereus (Gram-positive bacteria), Escherichia coli, Pseudomonas aeruginosa (Gram-negative bacteria); and Candida albicans (Fungi). Compounds 3 and 4 showed stronger inhibition of Gram-positive bacteria and fungi growth, and the antimicrobial ability of compound 3j (a 4-(benzylamino)-5-chloro-2,6-difluoro analog, MIC[SA] = 0.5 µg/mL; MIC[BC] = 0.4 µg/mL; MIC[CA] = 0.5 µg/mL) were close to nofloxacin and fluconazole and identified as the most potent antimicrobial agents in the series. The preliminary analysis of structure-activity relationships is also discussed.

Cloning, bioinformatics and the enzyme activity analyses of a phenylalanine ammonia-lyase gene involved in dragon's blood biosynthesis in Dracaena cambodiana.

Phenylalanine ammonia-lyase (PAL) is the key enzyme of the phenylpropanoid pathway, playing an important role in plant development and defence. We cloned a partial cDNA of PAL gene, DcPAL1, from Dracaena cambodiana seedlings using RT-PCR with degenerate primers that were designed based on a multiple sequence alignment of known PAL genes from other plant species. DcPAL1 shows highly homologous to other known PAL genes registered in GenBank, being closest to that of Musa acuminata. DcPAL1 has a relatively high GC content and most of the GC is in the third codon position. It has 768 bp in size with a maximum open reading frame (ORF) of 765 bp, encoding a 255 amino acid-polypeptide. The deduced PAL protein is a stable protein, having classical PAL domains and consisting of three major hydrophobic domains. Analysis of effective number of codons (ENC) shows that DcPAL1 codons are used at equal frequency. Relatively higher usage frequency appears randomly in codons ended with any of the four bases; six codons have no usage bias. There are 45 codons showing distinct usage preference between DcPAL1 and E. coli, 20 between DcPAL1 and yeast. Therefore, the yeast system may be more suitable for the expression of DcPAL1. Upon the elicitation of Fusarium proliferatum, a potent elicitor of dragon's blood, the PAL enzyme activity in the leaves and stems of D. cambodiana and other two Dracaena spp. significantly increased, accompanying with the formation of dragon's blood, indicating the involvement of PAL in the biosynthesis of dragon's blood, a precious traditional medicine.

Seasonal variation for the ratio of BaP to BeP at different sites in Great Xiamen Bay.

From March 2008 to February 2009, PM(10) samples were collected and analyzed for polycyclic aromatic hydrocarbons (PAHs) at eight sampling sites in Great Xiamen Bay, China. Analyses of the seasonal and spatial variations of these compounds revealed the following results. Significantly high levels of PAHs were found in the winter compared to the summer, sometimes exceeding 100 ng m(-3), and the spatial variations were influenced most by the sampling site surroundings. Composition profiles of PAHs of an urban and a rural site were shown to be very similar with a positive correlation coefficient larger than 0.9 at the 0.01 level of significance for the same season. Diagnostic ratios, together with principal component and multiple linear regression analysis, showed that more PAHs were from grass/wood/coal combustion in winter than in other seasons. The ratios of benzo[a]pyrene to benzo[e]pyrene (BaP-BeP) in winter and fall were 0.6-1.7 times higher than those in spring and summer, suggesting the importance of local emissions of PAHs. The BaP-BeP ratios in Kinmen were generally lower than those in Xiamen, indicating that the aging degree of PAHs was higher in Kinmen than in Xiamen. The external input of PAHs from upwind urban and industrial areas was one of the key factors causing high levels of PAHs in PM(10) in Great Xiamen Bay in winter.

Proposal of Intrasporangium mesophilum sp. nov., and reclassification of Humihabitans oryzae Kageyama et al. 2007 as Intrasporangium oryzae comb. nov.

A gram-positive and aerobic bacterium, designated strain YIM 49065(T), was isolated from rhizospheric soil of Jatropha curcas in Yunnan, China. This isolate formed branched and fragmented mycelia containing ll-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The major cellular fatty acid profile was characterized by iso-C(14 : 0), iso-C(15 : 0) and iso-C(16 : 0), and the predominant menaquinone was MK-8(H(4)). The DNA G+C content was 69.6 mol%. A phylogenetic analysis based on 16S rRNA gene sequence demonstrated that strain YIM 49065(T) was closely associated with Intrasporangium calvum DSM 43043(T) and Humihabitans oryzae KV-657(T), exhibiting 98.8 % and 98.6 % 16S rRNA gene sequence similarities, respectively. Furthermore, DNA-DNA hybridizations among strains YIM 49065(T), I. calvum DSM 43043(T) and H. oryzae DSM 22331(T) indicated that this isolate represented a novel species in the genus Intrasporangium. On the basis of these data, we proposed one novel species, Intrasporangium mesophilum sp. nov., for strain YIM 49065(T) ( = DSM 23217(T)  = CCTCC AA 209077(T)). Additionally, the comparison of biochemical and phylogenetic characters supported the reclassification of Humihabitans oryzae as a separate species within the genus Intrasporangium, Intrasporangium oryzae comb. nov. (type strain KV-657(T)  = JCM 15924(T)  = NBRC 101802(T)  = NRRL B-24470(T)).

Production of dragon's blood in Dracaena cochinchinensis plants by inoculation of Fusarium proliferatum.

Dragon's blood is a traditional medicine widely used in the world from ancient times. However, little is known about its formation mechanism. This work aimed to gain some insights into its formation mechanism and to control its production. The results demonstrate that wounding plus causal fungal infection and keeping the wound moist are essential for efficient dragon's blood formation in Dracaena cochinchinensis. Two fungal isolates YM-266 and YM-71213 of Fusarium proliferatum increased the yield of dragon's blood in D. cochinchinensis trees by 2.7- and 3.3-times compared to that of the control (wounding alone and keeping the wound moist), respectively. The fungal induced dragon's blood had almost identical chemical constituents to that of the natural dragon's blood with a higher loureirins a and b content as analyzed by TLC and HPLC. In addition, the induced dragon's blood had similar antimicrobial activity and similar or higher antioxidant activity than that of the natural dragon's blood. The novel biological technology developed here for the production of dragon's blood is safe, repeatable, practical, and feasible for the farmers, enabling the production of dragon's blood in a sustainable way without destroying the endangered trees and environment.

Morphological characteristics of spermatozoa before and after renal transplantation.

AIM: To investigate the changes of the spermatozoa ultrastructures before and after renal transplantation in uremic patients. METHODS: The sperm of five uremic patients before and after transplantation and four healthy volunteers were collected and examined by scanning electron microscopy. RESULTS: Abnormal spermatozoa were found in patients pre-transplantation; abnormalities included deletion of the acrosome, absence of the postacrosomal and postnuclear ring, dumbbell-like changes of the head, tail curling, and absence of the mitochondrial sheath in the mid-segment. After renal transplantation, most of the spermatozoa became normal. CONCLUSION: There are many abnormalities with regard to the appearance and structure of the head, acrosome, mitochondria and tail of the spermatozoa in uremic patients. The majority of the spermatozoa returned to normal after renal transplantation, but a few still presented some abnormalities possibly relating to the administration of immunosuppressants.