Egicoccus halophilus gen. nov., sp. nov., a halophilic, alkalitolerant actinobacterium and proposal of Egicoccaceae fam. nov. and Egicoccales ord. nov.

A novel Gram-stain-positive, non-motile, moderately halophilic and alkalitolerant actinobacterium, designated EGI 80432T, was isolated from a saline-alkaline soil of Xinjiang province, north-west China. Cells were non-endospore-forming cocci with a diameter of 0.5-0.8 µm. Strain EGI 80432T grew in the presence of 0-9 % (w/v) NaCl (optimum at 3-5 %), and also grew within the pH range 6.0-10.0 (optimum at pH 8.0-9.0) on marine 2216E medium. The peptidoglycan type was A1γ. The whole-cell hydrolysates contained glucose, galactose, mannose and three unknown sugars as major sugars. The predominant menaquinone was MK-9(H4). The major fatty acids were C17 : 1ω8c, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), C18 : 1ω9c and iso-C15 : 0 The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, one unknown phosphoglycolipid, three unknown phospholipids and four unknown polar lipids. The genomic DNA G+C content was 75.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain EGI 80432T clustered within the radius of the class Nitriliruptoria. Levels of sequence similarity between strain EGI 80432T and its phylogenetic neighbours Nitriliruptor alkaliphilus ANL-iso2T and Euzebya tangerina F10T were 94.1 and 88.1 %, respectively. Based on morphological, physiological and chemotaxonomic characteristics and phylogenetic analysis, a novel species of a new genus, Egicoccus halophilus gen. nov., sp. nov., is proposed, within the new family and new order Egicoccaceae fam. nov. and Egicoccales ord. nov. in the class Nitriliruptoria. The type strain of Egicoccus halophilus is EGI 80432T ( = CGMCC 1.14988T = KCTC 33612T).

Okibacterium endophyticum sp. nov., a novel endophytic actinobacterium isolated from roots of Salsola affinis C. A. Mey.

A white bacterial strain, designated EGI 650022(T), was isolated from the roots of Salsola affinis C. A. Mey, collected from Urumqi City, Xinjiang, north-western China. The strain was found to be aerobic, Gram-stain positive, oxidase-positive and catalase-positive. Cells were non-motile and irregular rods. Growth occurred at NaCl concentrations between 0 and 7 % (w/v), temperatures between 5 and 45 °C, and pH 6.0-9.0. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain EGI 650022(T) belongs to a clade with the genera Okibacterium and Plantibacter in the family Microbacteriaceae. The novel strain EGI 650022(T) showed highest levels of 16S rRNA gene sequence similarity with members of the genera Okibacterium and Plantibacter (97.2-98.0 %). The cell-wall peptidoglycan contained glutamate, homoserine, glycine, alanine and lysine. The predominant menaquinones (MKs) were MK-11, MK-12 and MK-12 (H4). The polar lipid pattern comprised phosphatidylglycerol, diphosphatidylglycerol, two unknown glycolipids and two unknown phospholipids. The major fatty acids were anteiso-C15:0 and anteiso-C17:0. The DNA G+C content was 66.0 mol%. The DNA-DNA relatedness values of strain EGI 650022(T) with Okibacterium fritillariae DSM 12584(T), Plantibacter flavus DSM 14012(T) and Plantibacter auratus DSM 19586(T) were 39.7, 19.7 and 22.0 %. Based on phylogenetic, phenotypic, chemotaxonomic and DNA-DNA hybridization data, strain EGI 650022(T) is considered to represent a novel species of the genus Okibacterium, for which the name Okibacterium endophyticum sp. nov. is proposed; the type strain is EGI 650022(T) (=JCM 30086(T) = KCTC 29492(T)).

Frigoribacterium endophyticum sp. nov., an endophytic actinobacterium isolated from the root of Anabasis elatior (C. A. Mey.) Schischk.

A novel endophytic actinobacterium, designated EGI 6500707(T), was isolated from the surface-sterilized root of a halophyte Anabasis elatior (C. A. Mey.) Schischk collected from Urumqi, Xinjiang province, north-west China, and characterized using a polyphasic approach. Cells were Gram-stain-positive, non-motile, short rods and produced white colonies. Growth occurred at 10-45 °C (optimum 25-30 °C), at pH 5-10 (optimum pH 8) and in presence of 0-4% (w/v) NaCl (optimum 0-3%). The predominant menaquinone was MK-9. The diagnostic phospholipids were diphosphatidylglycerol and phosphatidylglycerol. The major fatty acids were anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0). The DNA G+C content of strain EGI 6500707(T) was 69.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain EGI 6500707(T) should be placed in the genus Frigoribacterium (family Microbacteriaceae , phylum Actinobacteria ), and that the novel strain exhibited the highest 16S rRNA gene sequence similarity to Frigoribacterium faeni JCM 11265(T) (99.1%) and Frigoribacterium mesophilum MSL-08(T) (96.5%). DNA-DNA relatedness between strain EGI 6500707(T) and F. faeni JCM 11265(T) was 47.2%. On the basis of phenotypic and chemotaxonomic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain EGI 6500707(T) represents a novel species of the genus Frigoribacterium , for which the name Frigoribacterium endophyticum sp. nov. is proposed. The type strain is EGI 6500707(T) ( = JCM 30093(T) = KCTC 29493(T)).

Labedella endophytica sp. nov., a novel endophytic actinobacterium isolated from stem of Anabasis elatior (C. A. Mey.) Schischk.

A Gram-stain positive, yellow-coloured, aerobic, non-motile, catalase-positive and oxidase-negative, endophytic actinobacterium, designated strain EGI 6500705(T), was isolated from the surface-sterilized stem of a halophyte Anabasis elatior (C. A. Mey.) Schischk collected from Urumqi, Xinjiang province, north-west China. The organism had ornithine as the diagnostic cell-wall diamino acid. The major fatty acids were anteiso-C15:0, anteiso-C17:0 and iso-C16:0. The predominant menaquinones were MK-9, MK-10 and MK-11. The DNA G + C content of strain EGI 6500705(T) was 69.2 mol%. Phylogenetic analyses based on 16S rRNA gene sequence showed that strain EGI 6500705(T) is clearly affiliated with the genus Labedella and most closely related to Labedella gwakjiensis KCTC 19176(T), with 99.0 % sequence similarity. DNA-DNA relatedness between strain EGI 6500705(T) and L. gwakjiensis KCTC 19176(T) was 27.4 %. On the basis of phenophytic, chemotaxonomic and phylogenetic analysis, the strain EGI 6500705(T) represents a novel species of the genus Labedella, for which the name Labedella endophytica sp. nov. is proposed. The type strain is EGI 6500705(T) (=KCTC 29494(T) = CPCC 203961(T) = JCM 30092(T)).

Nesterenkonia rhizosphaerae sp. nov., an alkaliphilic actinobacterium isolated from rhizosphere soil in a saline-alkaline desert.

An alkaliphilic actinobacterial strain, designated EGI 80099(T), was isolated from a rhizosphere soil sample of Reaumuria soongorica found in the desert soils of Fukang, Xinjiang, north-west China. Cells of strain EGI 80099(T) were Gram-stain-positive, non-motile, non-endospore-forming cocci. The predominant menaquinones were MK-7, MK-8 and MK-9. The major cellular fatty acids (>10 %) were anteiso-C15 : 0 and anteiso-C17 : 0. Analysis of the cell wall showed the presence of peptidoglycan of the type l-Lys-Gly-l-Glu, variation A4α. Cells of the isolate contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, an unknown phospholipid and an unidentified glycolipid as polar lipids. The genomic DNA G+C content was 63.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EGI 80099(T) belongs to the genus Nesterenkonia, sharing 95.68-97.37 % sequence similarities with the type strains of recognized species within this genus. DNA-DNA hybridization of strain EGI 80099(T) with the type strains of species that showed the highest sequence similarities, Nesterenkonia aethiopica DSM 17733(T) (97.37 %), Nesterenkonia flava CAAS 251(T) (97.23 %) and Nesterenkonia xinjiangensis YIM 70097(T) (97.02 %), gave relatedness values of 8.7-62.2 %. Data from DNA-DNA hybridizations and physiological and biochemical tests indicated that strain EGI 80099(T) represents a novel species of the genus Nesterenkonia, for which the name Nesterenkonia rhizosphaerae sp. nov. is proposed. The type strain is EGI 80099(T) ( = BCRC 16947(T) = JCM 19129(T)).

The complete mitochondrial genomes of two ghost moths, Thitarodes renzhiensis and Thitarodes yunnanensis: the ancestral gene arrangement in Lepidoptera.

BACKGROUND: Lepidoptera encompasses more than 160,000 described species that have been classified into 45-48 superfamilies. The previously determined Lepidoptera mitochondrial genomes (mitogenomes) are limited to six superfamilies of the lineage Ditrysia. Compared with the ancestral insect gene order, these mitogenomes all contain a tRNA rearrangement. To gain new insights into Lepidoptera mitogenome evolution, we sequenced the mitogenomes of two ghost moths that belong to the non-ditrysian lineage Hepialoidea and conducted a comparative mitogenomic analysis across Lepidoptera. RESULTS: The mitogenomes of Thitarodes renzhiensis and T. yunnanensis are 16,173 bp and 15,816 bp long with an A + T content of 81.28 % and 82.34 %, respectively. Both mitogenomes include 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and the A + T-rich region. Different tandem repeats in the A + T-rich region mainly account for the size difference between the two mitogenomes. All the protein-coding genes start with typical mitochondrial initiation codons, except for cox1 (CGA) and nad1 (TTG) in both mitogenomes. The anticodon of trnS(AGN) in T. renzhiensis and T. yunnanensis is UCU instead of the mostly used GCU in other sequenced Lepidoptera mitogenomes. The 1,584-bp sequence from rrnS to nad2 was also determined for an unspecified ghost moth (Thitarodes sp.), which has no repetitive sequence in the A + T-rich region. All three Thitarodes species possess the ancestral gene order with trnI-trnQ-trnM located between the A + T-rich region and nad2, which is different from the gene order trnM-trnI-trnQ in all previously sequenced Lepidoptera species. The formerly identified conserved elements of Lepidoptera mitogenomes (i.e. the motif 'ATAGA' and poly-T stretch in the A + T-rich region and the long intergenic spacer upstream of nad2) are absent in the Thitarodes mitogenomes. CONCLUSION: The mitogenomes of T. renzhiensis and T. yunnanensis exhibit unusual features compared with the previously determined Lepidoptera mitogenomes. Their ancestral gene order indicates that the tRNA rearrangement event(s) likely occurred after Hepialoidea diverged from other lepidopteran lineages. Characterization of the two ghost moth mitogenomes has enriched our knowledge of Lepidoptera mitogenomes and contributed to our understanding of the mechanisms underlying mitogenome evolution, especially gene rearrangements.

[Prognostic analysis of stage III-IV non-small cell lung cancer patients treated by traditional chinese medicine].

BACKGROUND & OBJECTIVE: Chemotherapy is a treatment for stage III-IV non-small cell lung cancer (NSCLC), but the efficacy is not ideal. Traditional Chinese medicine (TCM) has certain effect on NSCLC. This study was to investigate various factors that affect the prognosis of advanced NSCLC, and evaluate the role of TCM in enlonging survival time of patients with stage III-IV NSCLC. METHODS: The NSCLC patients who meet the inclusive criteria were randomized into TCM group, combination (TCM plus NP regimen) group, and chemotherapy group, and received relevant treatments. The median survival time (MST) was calculated by Kaplan-Meier method. The prognosis of the patients was analyzed by COX regression method. RESULTS: A total of 294 stage III-IV NSCLC patients were enrolled, of which 99 were in TCM group, 103 in combination group, 92 in chemotherapy group. The MST were 292 days in TCM group, 355 days in combination group, and 236 days in chemotherapy group; the cumulative survival rates were 45.38%, 48.86%, and 42.17%, respectively (P>0.05). Cox regression analysis indicated that therapy, gender, disease course, erythrocyte sedimentation, KPS score, tumor size, and patient's weight were independent prognostic factors of stage III-IV NSCLC. CONCLUSION: Compare with chemotherapy alone, TCM combined with chemotherapy may prolong the survival time of stage III-IV NSCLC patients.

[Tolerance of human body to simultaneous infusion of 5-fluorouracil and calcium folinate].

BACKGROUND & OBJECTIVE: It has been confirmed that calcium folinate (CF), as a chemotherapy-biochemical modulator of 5-fluorouracil (5-FU), can improve the effect of antitumor of 5-FU. But the usage and dosage of this therapy should be further studied. The purpose of this article was to study the maximum tolerated dose (MTD) and the dose limiting toxicity (DLT) of human body to simultaneously intravenous infusion of 5-FU and CF. METHODS: The cases being up to the selecting standard were treated with DDP+CF/5-FU regimen. The dose of CF and DDP for every course was fixed. The first dose of 5-FU was 500 mg x (m(2) x d)(-1),then the doses were increased by degree of 50 mg x (m(2) x d)(-1), if no severe side effect was observed, up to maximum toxicity tolerated dose. There were 3-6 cases in each degree of dose. RESULTS: Thirty-two cases were treated and given total 64 courses of the administration in all. The overall remission rate was 84.4%. The MTD of 5-FU was 800 mg x (m(2) x d)(-1). Mucositis and diarrhea were found to be the DLT. CONCLUSION: Simultaneous continuous infusion of 5-FU and CF can enhance the dosage of 5-FU, and has a better efficacy to advanced digestive tract, head and neck cancers. So the authors recommend that the dosage of 5-FU in phase II clinical trial is 700 mg x (m(2) x d)(-1). This dose can be used for 5 consecutive days and be repeated after 3 weeks.