Effects and mechanism of siomycin A on the growth and apoptosis of MiaPaCa-2 cancer cells.

Siomycin A is a type of thiopeptide antibiotic that is isolated from the fermentation products of an endophytic actinomycin, which is derived from the medicinal plant Acanthopanax senticosus. The present study investigated whether siomycin A has antitumor effects in vitro on a variety of cell lines. A Cell Counting Kit-8 assay was performed to detect the effects of siomycin A on cell viability; morphological changes in the MiaPaCa-2 cell line were analyzed using an inverted phase contrast microscope. A Transwell migration assay was applied to detect cell migration ability. The cytoskeleton was observed by laser confocal microscopy, and apoptosis was detected using flow cytometry. A western blot assay was used to detect the expression of matrix metalloproteinase (MMP)-2, MMP-9 and α-tubulin. The results revealed that siomycin A inhibited the proliferation of human tumor cell lines of different origins. As the concentration of siomycin A increased, the cell density decreased gradually and cells exhibited a morphological change from spindle to spherical shape. Furthermore, 24 h after administration, the cell migration ability was inhibited. The cytoskeleton complexity and morphological changes were increased after administration of siomycin A. The percentage of apoptotic cells was significantly increased and the expression levels of MMP-2, MMP-9 and α-tubulin were downregulated by siomycin A. Therefore, siomycin A was determined to effectively inhibit the proliferative ability of a variety of human tumor cell lines. Siomycin A was also determined to affect the cytoskeleton of tumor cells by downregulating the expression of α-tubulin protein.

Effect of exogenous pulmonary surfactants on mortality rate in neonatal respiratory distress syndrome: A network meta-analysis of randomized controlled trials.

BACKGROUND: The utilization of multiple natural and synthetic products in surfactant replacement therapies in treatment of neonatal respiratory distress syndrome (NRDS) prompted us to take a closer looks at these various therapeutic options and their efficacies. The purpose of our study was to evaluate the effects of six exogenous pulmonary surfactants (EPS) (Survanta, Alveofact, Infasurf, Curosurf, Surfaxin and Exosurf) on mortality rate in NRDS by a network meta-analysis. METHODS: An exhaustive search of electronic databases was performed in PubMed, Ovid, EBSCO, Springerlink, Wiley, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang and VIP databases (last updated search in October 2014) to retrieve randomized controlled trials (RCTs) relevant to our study topic. Published clinical trials were screened based on the following inclusion criteria: (1) study design: RCTs; (2) interventions: treatment with Survanta, Alveofact, Infasurf, Curosurf, Surfaxin or Exosurf for NRDS; (3) study subject: infants with NRDS confirmed by clinical diagnosis; (4) outcome: the mortality rate of infants with NRDS. Statistical analysis was performed using Stata 12.0 software (Stata Corporation, College Station, TX, USA) and Comprehensive Meta-analysis (CMA 2.0) software. RESULTS: From the 1840 studies initially retrieved through database searches, a total of 17 high quality RCTs were selected for this network meta-analysis. The selected studies included a combined total of 57,223 infants with NRDS treated with various EPS (Survanta, 27,017; Alveofact, 159; Infasurf, 20,377; Curosurf, 20,911; Surfaxin, 646; Exosurf, 1640). Network meta-analysis results showed that the mortality rates in NRDS infants treated with Alveofact, Infasurf, Curosurf, Surfaxin, Exosurf were not significantly different compared to Survanta (Alveofact: OR = 1.163, 95% CI = 0.645-2.099, P = 0.616; Infasurf: OR = 0.985, 95% CI = 0.777-1.248, P = 0.897; Curosurf: OR = 0.789, 95% CI = 0.619-1.007, P = 0.056; Surfaxin: OR = 0.728, 95% CI = 0.477-1.112, P = 0.142; Exosurf: OR = 0.960, 95% CI = 0.698-1.319, P = 0.799). Notably, the surface under the cumulative ranking curves (SUCRA) value in Surfaxin group was significantly higher than the other five groups (Surfaxin: 80.4%; Survanta: 37.0%; Alveofact: 24.4%; Infasurf: 40.0%; Curosurf: 73.9%; Exosurf: 44.2%), suggesting that infant mortality rate in Surfaxin group was the lowest among the six EPS groups. CONCLUSION: Our study demonstrated that Surfaxin could effectively reduce the mortality rate of infants with NRDS and may have a better efficacy in NRDS treatment, compared to Survanta, Alveofact, Infasurf, Curosurf and Exosurf.

Autophagy regulates hyperoxia-induced intracellular accumulation of surfactant protein C in alveolar type II cells.

Surfactant protein C (SP-C) deficiency is a risk factor for hyperoxia-induced bronchopulmonary dysplasia in newborn infants. However, the role of SP-C deficiency in the process is unclear. Here, using neonatal rat BPD model and MLE-12, mouse alveolar epithelial type II cell, we examined the changes of SP-C levels during hyperoxia. Immunohistochemistry, immunofluorescence, and ELISA analysis showed SP-C accumulation in alveolar epithelial type II cells. Electron microscopy further demonstrated the accumulation of lamellar bodies and the co-localization of lamellar bodies with autophagosomes in the cytoplasm of alveolar epithelial type II cells. The inhibition of autophagy with 3-Methyladenine and knockdown of Atg7 abolished hyperoxia-induced SP-C accumulation in the cytoplasm. Furthermore, inhibition of JNK signaling with SP600125 suppressed hyperoxia-induced Atg7 expression and SP-C accumulation. These findings suggest that hyperoxia triggers autophagy via JNK signaling-mediated Atg7 expression, which promotes the accumulation of SP-C within alveolar epithelial type II cells. Our data provide a potential approach for hyperoxic lung injury therapy by targeted pharmacological inhibition of autophagic pathway.

The role of placenta growth factor in the hyperoxia-induced acute lung injury in an animal model.

Prolonged exposure to hyperoxia leads to acute lung injury. Alveolar type II cells are main target of hyperoxia-induced lung injury. However, the cellular and molecular mechanisms remain unknown. Here, we aimed to investigate the role of placental growth factor (PLGF) in hyperoxia-induced lung injury. Using experimental hyperoxia-induced lung injury model of neonatal rat and mouse lung epithelial type II cells (MLE-12), we examined the levels of PLGF in bronchoalveolar lavage fluid and in the supernatants of MLE-12 cells. Our results revealed that exogenous PLGF induced hyperoxia-induced lung injury. Furthermore, PLGF triggered a shift of vinculin from insoluble to soluble cell fraction, similar to the observation under hyperoxia stimulation. Moreover, we observed significantly reduced phosphorylation of focal adhesion kinase and increased permeability in MLE-12 cells treated with PLGF. These results suggest that PLGF triggers focal adhesion disassembly in alveolar type II cells via inhibiting the activation of focal adhesion kinase. Our findings reveal a novel role of PLGF in hyperoxia-induced lung injury and provide a potential target for the management of hyperoxia-induced acute lung injury.

Morphological and molecular identification of two strains of dermatophytes.

In this study, we used traditional morphological and molecular identification methods to preliminarily identify two strains of dermatophytes. The two strains were observed under the microscope. And then the dermatophytes were cultured on Sabouraud's dextrose agar (SDA). The 18S rRNA regions of the two dermatophyte strains were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced and compared with GenBank data. BLAST tools and DNAMAN software were used to analyze the sequences. To further determine highly homologous sequences, a phylogenetic tree was constructed using the Neighbor-Joining method. The two strains of dermatophytes were identified by traditional morphological identification as Epidermophyton floccosum and Microsporum ferrugineum. The 18S rRNA sequence analyses showed high similarities to Cladosporium cladosporioides isolate C115LM-UFPR and Ascomycete sp. LB68A1A2. Epidermophyton and Cladosporium belong to dermatophyte, while Microsporum ferrugineum and Ascomycete belong to microsporum. The two novel strains of dermatophytes were therefore identified as Cladosporium cladosporioides isolate C115LM-UFPR (JN650537, Cladosporium) and Ascomycete sp. LB68A1A2 (AY770409, Ascomycete sp).

Diversity of culturable actinobacteria from Qinghai-Tibet plateau, China.

To investigate the diversity of culturable actinobacteria and further screen for microbial pharmaceutics, seven different media were chosen to isolate actinobacteria from 87 soil samples collected from Qinghai-Tibet plateau. A total of 1930 strains was isolated and identified to belong to 11 suborders, i.e., Actinopolysporineae, Corynebacterineae, Frankineae, Glycomycineae, Kineosporiineae, Micrococcineae, Micromonosporineae, Propionibacterineae, Pseudonocardineae, Streptomycineae and Streptosporangineae, and 16 families, i.e., Nocardioidaceae, Actinopolysporaceae, Actinosynnemataceae, Dermacoccaceae, Geodermatophilaceae, Glycomycetaceae, Kineosporiaceae, Microbacteriaceae, Micromonosporaceae, Nocardiaceae, Promicromonosporaceae, Propionibacteriaceae, Pseudonocardiaceae, Streptomycetaceae, Streptosporangiaceae and Thermomonosporaceae. A primary taxonomic study showed that at least 22 genera of actinobacteria were identified from the soil samples, among which ten isolates represented hitherto unknown species. The results showed that there was abundant actinobacterial species diversity in the soil samples from the Qinghai-Tibet plateau.

Actinopolymorpha cephalotaxi sp. nov., a novel actinomycete isolated from rhizosphere soil of the plant Cephalotaxus fortunei.

An actinomycete, strain I06-2230(T), was isolated from rhizosphere soil of the plant Cephalotaxus fortunei, collected from Yunnan province, south China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belongs to the genus Actinopolymorpha. Cells grew on agar surfaces, with no penetration even after prolonged cultivation. Aerial hyphae were absent. Cells were irregularly shaped and remained attached as chains or aggregates. Chemotaxonomic data, which showed ll-diaminopimelic acid in the cell wall, glucose as the whole-cell sugar, type PI phospholipids and MK-9(H4) as the predominant menaquinone, supported the affiliation of strain I06-2230(T) to the genus Actinopolymorpha. The major fatty acids were iso-C(15 : 0), iso-C(16 : 0) and iso-C(16 : 1) H. The genomic DNA G+C content was 69.3 mol%. DNA-DNA hybridization data, in combination with chemotaxonomic, physiological and biochemical data, demonstrated that strain I06-2230(T) should be classified as representing a novel species of the genus Actinopolymorpha. The name Actinopolymorpha cephalotaxi sp. nov. is proposed, with strain I06-2230(T) (=DSM 45117(T)=CCM 7466(T)=KCTC 19293(T)) as the type strain.

Alloactinosynnema album gen. nov., sp. nov., a member of the family Actinosynnemataceae isolated from soil.

The taxonomic position of a Gram-stain-positive, aerobic strain, designated 03-9939(T), isolated from a soil sample collected from Xinjiang Province, China, was established using a polyphasic approach. Whole-cell hydrolysates of strain 03-9939(T) contained galactose and ribose as diagnostic sugars and meso-diaminopimelic acid as the diamino acid. The predominant menaquinone was MK-9(H4). The phospholipids consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylcholine. The major fatty acids were iso-C(16 : 0) (61.5 %) and iso-C(16 : 1) H (11.6 %). The genomic DNA G+C content was 68.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 03-9939(T) should be placed within the family Actinosynnemataceae, in which the strain formed a distinct lineage. Signature nucleotides in the 16S rRNA gene sequence showed that the strain contained a genus-specific diagnostic nucleotide signature pattern. The combination of phylogenetic analysis, phenotypic characteristics and chemotaxonomic data supported the conclusion that strain 03-9939(T) represents a novel species in a new genus of the family Actinosynnemataceae, for which the name Alloactinosynnema album gen. nov., sp. nov. is proposed. Strain 03-9939(T) (=DSM 45114(T) =KCTC 19294(T) =CCM 7461(T)) is the type strain of Alloactinosynnema album.

Saccharopolyspora antimicrobica sp. nov., an actinomycete from soil.

Three Gram-positive, aerobic, non-motile, non-acid-alcohol-fast strains, designated I05-00051, I05-00074T and I03-00808, were isolated from different soil samples in Beijing and Sichuan, China. Phylogenetic analysis based on 16S rRNA gene sequences and DNA-DNA hybridization experiments revealed that these three isolates represented the same genospecies. These three strains showed <97.0 % 16S rRNA gene sequence similarity with the type strains of recognized species of the genus Saccharopolyspora, with the exception of Saccharopolyspora hirsuta subsp. hirsuta DSM 43463T (98.1 % gene sequence similarity) and Saccharopolyspora spinosa DSM 44228T (98.0 % similarity). Chemotaxonomic data, including meso-diaminopimelic acid as the diagnostic diamino acid, arabinose and galactose as predominant sugars, iso-C15 : 0, iso-C16 : 0, iso-C17 : 0 and anteiso-C17 : 0 as major fatty acids, MK-9(H4) as predominant menaquinone and polar lipids dominated by diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylinositol, supported the affiliation of these three organisms to the genus Saccharopolyspora. The genomic DNA G+C contents of the three isolates were 68.2-69.9 mol%. The results of DNA-DNA hybridization experiments among these three isolates and S. hirsuta subsp. hirsuta DSM 43463T and S. spinosa DSM 44228T, in combination with chemotaxonomic and physiological data, demonstrated that the three new isolates represent a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora antimicrobica sp. nov. is proposed. The type strain is I05-00074T (=CCM 7463T=KCTC 19303T).

The disturbance of small RNA pathways enhanced abscisic acid response and multiple stress responses in Arabidopsis.

The phytohormone abscisic acid (ABA) regulates plant growth and development as well as stress tolerance. To gain more insights into ABA signalling, a population of chemical-inducible activation-tagged Arabidopsis mutants was screened on the basis of the ABA effect on the inhibition of seed germination. Two novel ABA supersensitive mutants ABA supersensitive during germination1 (absg1) and absg2 were characterized as alleles of Dicer-like1 (DCL1) and HEN1, respectively, as microRNA biogenesis genes, and accordingly, these two mutants were renamed dcl1-11 and hen1-16. The dcl1-11 mutant was an ABA hypersensitive mutant for seed germination and root growth. Reverse transcriptase polymerase chain reaction assays revealed that the expression of ABA- and stress-responsive genes was increased in dcl1-11, as compared with the wild type (WT). Furthermore, the germination assay showed that dcl1-11 was also more sensitive to salt and osmotic stress. The hen1-16 mutant also showed supersensitive to ABA during seed germination. Further analysis showed that, among the microRNA biogenesis genes, all the other mutants were not only enhanced in sensitivity to ABA, salt and osmotic stress, but also enhanced the expression of ABA-responsive genes. In addition to the mutants in the microRNA biogenesis, the interruption of the production of crucial components of other small RNA pathways such as dcl2, dcl3 and dcl4 also caused ABA supersensitive during germination.

[RAPD analysis on different isolates of Trichomonas vaginalis].

OBJECTIVE: To study genetic polymorphism of DNA on seven isolates of Trichomonas vaginalis. METHODS: The random amplified polymorphic DNA (RAPD) technique was performed to amplify genomic DNA of the seven T. vaginalis isolates, including Beijing 1, Beijing 2, Chengde, Tangshan, Jiujiang 1, Jiujiang 2 and Jiujiang 3. The DNA bands detected were analyzed by clustering analysis with SPSS software. RESULTS: The percentage of genetic similarity among the seven isolates was from 77.4% to 94.7%, showing a close genetic relationship among them. The percentages between the isolates of Beijing 1 and Tangshan, Jiujiang 1 and Jiujiang 2, Beijing 2 and Jiujiang 3 were 89.2%, 92.1% and 94.7% respectively, while that of Jiujiang 1 and Chengde was 77.4%, indicating a lower homology. CONCLUSION: There are a close genetic relationship and certain gene polymorphism among the seven T. vaginalis isolates; geographical origin plays little role to the genetic characteristics.

[Study on surface adhesion protein 33 gene sequence of different Trichomonas vaginalis isolates].

OBJECTIVE: To study genetic polymorphism of surface adhesion protein 33 (AP33) gene on the seven isolates of Trichomonas vaginalis. METHODS: PCR technique was performed to amplify AP33 gene from the seven isolates, DNA sequences were obtained from the AP33 gene of the isolates and phylogenetic tree was built. Minimal lethal concentrations (MLC) of metronidazole on the isolates were measured in vitro. RESULTS: Percentage of the similarity between 7 isolates and U87098 in GenBank was 98.2%-100%, which indicated a high homology and belonged to isotype isolates. There were four branches between Beijing 1 isolate and Tangshan isolate, Jiujiang 1 isolate and Jiujiang 2 isolate, Beijing 2 isolate and Jiujiang 3 isolate, Chengde isolate and U87098 isolate in phylogenetic tree, which showed a close genetic relationship respectively. No relativity was detected between geographical origin and genetic relationship. CONCLUSION: There is a close genetic relationship among the seven T. vaginalis isolates. MLC showed a difference between isolates which have close relationship.

[Isoenzyme analysis on different isolates of Trichomonas vaginalis].

OBJECTIVE: To study the biological types on the seven isolates of Trichomonas vaginalis from Beijing, Hebei-Tangshan, Hebei-Chengde and Jiangxi-Jiujiang in the mainland of China. METHODS: The samples were analyzed by PAGE, isoenzyme stain and cluster analysis. RESULTS: The isoenzyme systems used in the study included MDH, LDH, G-6-PD, PGI and PGM. No difference in the isoenzyme patterns of G-6-PD and PGI was found among the seven isolates. The MDH and LDH patterns of Beijing 1, Beijing 2, Jiujiang 3 strains were identical, while they were distinguishable from those of Chengde, Tangshan, Jiujiang 1, Jiujiang 2 isolates. The PGM pattern of Beijing 1 and Beijing 2 isolates were same but was different from that of the remainders. Gene tree was constructed according to the isoenzyme profiles. The results showed that there are differences in the patterns of the five isoenzymes between the isolates of Beijing 1, Beijing 2, Jiujiang 3 and other four isolates, and Jiujiang 3 was different from Beijing 1, Beijing 2 slightly. CONCLUSION: It seems reasonable to assume that there are at least three different biological types of Trichomonas vaginalis in China.