Safety of embryo cryopreservation: insights from mid-term placental transcriptional changes.

BACKGROUND: In recent years, with benefits from the continuous improvement of clinical technology and the advantage of fertility preservation, the application of embryo cryopreservation has been growing rapidly worldwide. However, amidst this growth, concerns about its safety persist. Numerous studies have highlighted the elevated risk of perinatal complications linked to frozen embryo transfer (FET), such as large for gestational age (LGA) and hypertensive disorders during pregnancy. Thus, it is imperative to explore the potential risk of embryo cryopreservation and its related mechanisms. METHODS: Given the strict ethical constraints on clinical samples, we employed mouse models in this study. Three experimental groups were established: the naturally conceived (NC) group, the fresh embryo transfer (Fresh-ET) group, and the FET group. Blastocyst formation rates and implantation rates were calculated post-embryo cryopreservation. The impact of FET on fetal growth was evaluated upon fetal and placental weight. Placental RNA-seq was conducted, encompassing comprehensive analyses of various comparisons (Fresh-ET vs. NC, FET vs. NC, and FET vs. Fresh-ET). RESULTS: Reduced rates of blastocyst formation and implantation were observed post-embryo cryopreservation. Fresh-ET resulted in a significant decrease in fetal weight compared to NC group, whereas FET reversed this decline. RNA-seq analysis indicated that the majority of the expression changes in FET were inherited from Fresh-ET, and alterations solely attributed to embryo cryopreservation were moderate. Unexpectedly, certain genes that showed alterations in Fresh-ET tended to be restored in FET. Further analysis suggested that this regression may underlie the improvement of fetal growth restriction in FET. The expression of imprinted genes was disrupted in both FET and Fresh-ET groups. CONCLUSION: Based on our experimental data on mouse models, the impact of embryo cryopreservation is less pronounced than other in vitro manipulations in Fresh-ET. However, the impairment of the embryonic developmental potential and the gene alterations in placenta still suggested it to be a risky operation.

Long distance signal transduction in response to environmental changes in plants.

The vascular system is responsible for the communication of information between different organs and the environment as a whole, so that it can coordinate the development of plants and respond to the changes of the environment. The signal substances moving in the vascular system are called long-distance signals. In recent years, it has been found that some long-distance molecular signals, such as microRNA, mRNA, small peptides, hormones, second messengers and proteins, can transmit extracellular stimuli from sensing tissues to target organs, so as to systematically regulate plant development process and environmental response. In this review, we summarize the molecular mechanisms of long-distance moving RNA, small peptides and proteins in plants to regulate plant organ development, nutrient uptake and stress resistance. The application potential of this field in crop breeding was discussed and prospected, in order to provide a theoretical basis for the application of genetics and breeding in crops.

Influence of bone anatomical morphology of mandibular molars on dental implant based on CBCT.

BACKGROUND: To apply CBCT to investigate the anatomical relationship between the mandibular molar and alveolar bone, aimed to provide clinical guidelines for the design of implant restoration. METHODS: 201 CBCT data were reevaluated to measure height of the alveolar process (EF), width of the alveolar process (GH), width of the basal bone (IJ), the angle between the long axis of the first molar and the alveolar bone (∠a) and the angle between the long axis of the alveolar bone and basal bone (∠b). The angle and width were measured to determine the implant-prosthodontic classification of the morphology in the left lower first molar (36) and right lower first molar (46). All measurements were performed on the improved cross-sectional images. RESULTS: EF, GH and IJ were measured as (10.83 ± 1.31) mm, (13.93 ± 2.00) mm and (12.68 ± 1.96) mm for 36, respectively; and (10.87 ± 1.24) mm, (13.86 ± 1.93) mm and (12.60 ± 1.90) mm for 46, respectively. No statistical significance was observed in EF, GH, IJ, ∠a and ∠b between 36 and 46 (all P > 0.05). The morphology was divided into three categories including the straight (68.7-69.2%), oblique (19.9-20.4%) and concave types (11%). Each type was consisted of two subcategories. CONCLUSIONS: The proposed classification could provide evidence for appropriate selection and direction design of the mandibular molar implant in clinical. The concave type was the most difficult to implant with the highest risk of lingual perforation. The implant length, width, direction required more attention.

Genetic improvement of nitrogen use efficiency in crops.

Nitrogen (N) is an essential mineral nutrient for plant growth and development. N deficiency is the major factor limiting plant growth and crop production in most natural and agricultural soils. The green revolution of the 1960's boosted crop yields through cultivation of semi-dwarf plant varieties. However, green revolution wheat and rice varieties have relatively poor nitrogen use efficiency (NUE), require a high N fertilizer supply to achieve maximum yield potential, and this leads to an increase in production costs and environmental problem. Therefore, a major challenge for sustainable agriculture is whether improvement of NUE through the reduction of N fertilizer supply can be achieved without yield penalty. In this review, we summarize the recent advances in understanding of molecular mechanisms underlying the regulation of N-responsive plant growth, utilization and possibility for improvements of NUE in crops, and new breeding strategies through modulation of N-responsive growth-metabolism coordination for future sustainable agriculture.

Premature Stroke Secondary to Severe Hypertension Results from Liddle Syndrome Caused by a Novel SCNN1B Mutation.

INTRODUCTION: Liddle syndrome (LS), an autosomal dominant and inherited monogenic hypertension syndrome caused by pathogenic mutations in the epithelial sodium channel (ENaC) genes SCNN1A, SCNN1B, and SCNN1G. OBJECTIVE: This study was designed to identify a novel SCNN1B missense mutation in a Chinese family with a history of stroke, and to confirm that the identified mutation is responsible for LS in this family. METHODS: DNA samples were collected from the proband and 11 additional relatives. Next-generation sequencing was performed in the proband to find candidate variants. In order to exclude genetic polymorphism, the candidate variantin SCNN1B was verified in other family members, 100 hypertensives, and 100 healthy controls by Sanger sequencing. RESULTS: Genetic testing revealeda novel and rare heterozygous variant in SCNN1B in the proband. This variant resulted in a substitution of threonine instead of proline at codon 617, altering the PY motif of ß-ENaC. The identified mutation was only verified in 5 relatives. In silico analyses indicated that this variant was highly pathogenic. In this family, phenotypic heterogeneity was present among 6 LS patients. Tailored medicine with amiloride was effective in controlling hypertension and improving the serum potassium concentration in patients with LS. CONCLUSIONS: We identified a novel SCNN1B mutation (c.1849C>A) in a family affected by LS. Patients with LS, especially those with severe hypertension, should be alert for the occurrence of premature stroke. Timely diagnosis using genetic testing and tailored treatment with amiloride can help LS patients to avoid severe complications.

The zinc-finger transcription factor BcMF20 and its orthologs in Cruciferae which are required for pollen development.

Brassica campestris Male Fertility 20 (BcMF20) is a typical zinc-finger transcription factor that was previously isolated from flower buds of Chinese cabbage (Brassica campestris ssp. chinensis). By applying expression pattern analysis, it can be known that BcMF20 was specifically and strongly expressed in tapetum and pollen, beginning from the uninucleate stage, and was maintained during the mature-pollen stage. As BcMF20 was highly conserved in Cruciferae, it can be indicated that this zinc-finger transcription factor is important during the growth of Cruciferae. In this study, 12 C2H2-type zinc-finger TFs which shared high homology with BcMF20 were found from NCBI via BLAST. A new molecular phylogenetic tree was constructed by the comparison between BcMF20 and these 12 C2H2-type zinc-finger TFs with NJ method. By analyzing this phylogenetic tree, the evolution of BcMF20 was discussed. Then, antisense RNA technology was applied in the transgenesis of Arabidopsis thaliana to get the deletion mutants of BcMF20, so that its function during the pollen development can be identified. The results showed: BcMF20 are in the same clade with three genes from Arabidopsis. The inhibition of BcMF20 expression led to smaller amounts of and lower rate in germination of pollen and lower rate in fruit setting in certain transgenetic plants. This also led to the complete collapse of pollen grains. By SEM and TEM, pollen morphology and anther development processes were observed. In the middle uninucleate microspore stage, a relatively thin or even no primexine was formed in microspores. This may result in the malformation of the pollen wall and finally cause the deformity of pollens. Above all, it can be indicated that BcMF20 may act as a part of regulation mechanisms of TAZ1 and MS1. Together they play a role in a genetic pathway in the tapetum to act on proliferation of tapetal cells and keep the normal development of pollens.

Conjugates of salicylaldoximes and peripheral site ligands: Novel efficient nonquaternary reactivators for nerve agent-inhibited acetylcholinesterase.

A new family of nonquaternary reactivators for nerve agent-inhibited human acetylcholinesterase (hAChE) were designed, synthesized and tested in this paper. It was found that salicylaldoximes were able to quickly cleave the P-S bond of organophosphate and avoid the reinhibition phenomenon in the reactivation process, but they lacked reactivating ability due to poor affinity for AChE. Based on a dual site binding strategy, different peripheral site ligands of AChE were introduced to achieve extra affinity. The in vitro reactivation experiments demonstrated that some of the yielding conjugates exhibited similar or even superior ability to reactivate sarin-, VX- or tabun-inhibited hAChE in comparison with the mono- and bis-pyridinium aldoximes currently used. Moreover, due to greatly improved lipophilicity, these nonquaternary conjugates hold promise for the development of efficient centrally activating reactivators.

Bioactive spirans and other constituents from the leaves of Cannabis sativa f. sativa.

In this paper, 17 compounds (1-17) were isolated from the leaves of Hemp (Cannabis sativa f. sativa). Among the isolates, two were determined to be new spirans: cannabispirketal (1), and α-cannabispiranol 4'-O-ß-D-glucopyranose (2) by 1D and 2D NMR spectroscopy, LC-MS, and HRESIMS. The known compounds 7, 8, 10, 13, 15, and 16 were isolated from Hemp (C. sativa f. sativa) for the first time. Furthermore, compounds 8 and 13 were isolated from the nature for the first time. All isolated compounds were evaluated for cytotoxicity on different tissue-derived passage cancer cell lines through cell viability and apoptosis assay. Among these compounds, compounds 5, 9 and 16 exhibited a broad-spectrum antitumor effect via inhibiting cell proliferation and promoting apoptosis. These results obtained have provided valuable clues to the understanding of the cytotoxic profile for these isolated compounds from Hemp (C. sativa f. sativa).

Glycocaulis alkaliphilus sp. nov., a dimorphic prosthecate bacterium isolated from crude oil.

A bacterial strain designated 6B-8(T) was isolated from crude oil from Daqing oilfield, China. Cells of strain 6B-8(T) were Gram-negative, aerobic, dimorphic and reproduced by means of binary fission. Strain 6B-8(T) could grow at 20-37 °C, pH 8-10 and 1-5 % (w/v) NaCl. Its genomic DNA G+C content was 62.0 mol%. The predominant cellular fatty acids were C18 : 1ω7c, C17 : 0, C18 : 0 and 11-methyl C18 : 1ω7c and the main hydroxy fatty acids were C12 : 0 3-OH and C12 : 1 3-OH when grown on marine agar 2216. The major quinone was Q-10 and the major polar lipids were three unidentified glycolipids. Phylogenetic analysis revealed that strain 6B-8(T) was a member of the family Hyphomonadaceae, sharing 99.6 and 99.4 % 16S rRNA gene sequence similarity with Glycocaulis abyssi LMG 27140(T) and Glycocaulis albus SLG210-30A1(T), respectively, and less than 94.4 % similarity with the type strains of other members of the family Hyphomonadaceae. However, the DNA-DNA relatedness between strain 6B-8(T) and related strains G. abyssi LMG 27140(T) and G. albus SLG210-30A1(T) was 36±5 and 42±5 %, respectively. In addition, several phenotypic and genotypic features allowed differentiation of strain 6B-8(T) from G. abyssi LMG 27140(T) and G. albus SLG210-30A1(T). Therefore, strain 6B-8(T) represents a novel species of genus Glycocaulis, for which the name Glycocaulis alkaliphilus sp. nov. is proposed. The type strain is 6B-8(T) ( = CGMCC 1.12428(T) = LMG 27410(T)).

Defluviimonas alba sp. nov., isolated from an oilfield.

Two Gram-stain-negative, rod-shaped bacterial strains, cai42T and b45, were isolated from oil-production water taken from Xinjiang Oilfield, China. Optimum growth was observed at 30 °C, at pH 8 and with 1-3% (w/v) NaCl. According to phylogenetic analyses, the two strains were members of the genus Defluviimonas, with 16S rRNA gene sequence similarities of 95.5-96.3 % with the type strains of species of the genus. The major cellular fatty acids of strains cai42T and b45 were C10 : 0 3-OH, C16 : 0 and summed feature 8 (C18 : 1ω7c/C18 : 1ω6c), and the predominant ubiquinone was Q-10, all of these data being typical for the genus Defluviimonas. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, glycolipid, phosphatidylcholine, two unidentified aminolipids, an unidentified phospholipid and two unidentified lipids. The mean genomic DNA G+C contents of strains cai42T and b45 were 60.8±1.1 and 60.4±1.0 mol%, respectively. On the basis of phylogenetic, physiological and chemotaxonomic analyses, strains cai42T and b45 represent a novel species of the genus Defluviimonas, for which the name Defluviimonas alba sp. nov. is proposed. The type strain is cai42T ( = CGMCC 1.12518T = LMG 27406T).

Halodurantibacterium flavum gen. nov., sp. nov., a non-phototrophic bacterium isolated from an oil production mixture.

Three Gram-negative bacterial strains, DQW12E6-69-1(T), DQW12E61-22-1, and DQW12E6-22-1-1, were isolated from an oil production mixture from Daqing Oilfield, northeastern China. The phylogenetic analysis based on the 16S rRNA gene sequences revealed that the three strains formed a stable cluster different from the known genus in Rhodobacteraceae of Alphaproteobacteria. In addition, they were most closely related to species in genera Pararhodobacter, Rhodobacter ,and Rhodobaca with the 16S rRNA gene sequence similarities being 95.1-95.9 %. Cells of the three strains were aerobic; they do not require salt to grow but are resistant to high salinity. They could conduct chemoorganoheterotrophic growth on various carbon sources, with non-phototrophic growth observed. The genomic DNA G+C contents of the strains DQW12E6-69-1(T), DQW12E6-22-1-1, and DQW12E61-22-1 were 63.8, 63.7, and 63.6 mol%, respectively. The predominant respiratory ubiquinone of DQW12E6-69-1(T) was Q-10, and the major fatty acids were C18:1 ω7c, C(18:0), and C(10:0) 3-OH. Photosynthetic pigments and photosynthetic reaction center gene pufM were not detected. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, unidentified glycolipid, and unidentified phospholipid. On the basis of phenotypic, genotypic, and chemotaxonomic characteristics, strains DQW12E6-69-1(T), DQW12E61-22-1, and DQW12E6-22-1-1 represent a novel genus and a novel species of the family Rhodobacteraceae. The name Halodurantibacterium flavum gen. nov., sp. nov. is proposed with strain DQW12E6-69-1(T) (=LMG 27742(T) = CGMCC 1.12756(T)) as the type strain.

Paradevosia shaoguanensis gen. nov., sp. nov., isolated from a coking wastewater.

A Gram staining negative, rod-shaped, aerobic bacterial strain J5-3(T) with a single polar flagellum was isolated from coking wastewater collected from Shaoguan, Guangdong, China. It was motile and capable of optimal growth at pH 6-8, 30 °C, and 0-2 % (w/v) NaCl. Its predominant fatty acids were 11-methyl C18:1 ω7c (29.2 %), C16:0 (20.6 %), C19:0 cyclo ω8c (18.2 %), C18:0 (11.0 %), and C18:1 ω7c/C18:1 ω6c (10.9 %) when grown on trypticase soy agar. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids (GL1, GL2), and two unknown phospholipid (PL1, PL2). The predominant ubiquinone was Q-10, and the genome DNA G+C content was 61.7 mol %. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain J5-3(T) belonged to the family Hyphomicrobiaceae in Alphaproteobacteria. It shared the 16S rRNA gene sequence similarities of 93.8-96.1 % with the genus Devosia, 94.5-94.8 % with the genus Pelagibacterium, and <92.0 % with all the other type strains in family Hyphomicrobiaceae. It can be distinguished from the closest phylogenetic neighbors based on several phenotypic and genotypic features, including α-galactosidase activity, tetracycline susceptibility, major fatty acid composition, polar lipid profile, DNA gyrase B subunit (gyrB) gene sequence, and random-amplified polymorphic DNA profile. Therefore, we consider strain J5-3(T) to represent a novel species of a novel genus within the family Hyphomicrobiaceae, for which the name Paradevosia shaoguanensis gen. nov., sp. nov. is proposed. The type strain of Paradevosia shaoguanensis is J5-3(T) (=CGMCC 1.12430(T) =LMG 27409(T)).

Plastorhodobacter daqingensis gen. nov., sp. nov.: a non-phototrophic bacterium isolated from Daqing Oilfield.

Two aerobic Gram staining negative, non-motile, and rod-shaped strains, DQW12E81-30(T) and DQW12E6-37-1, were isolated from an oil production mixture from Daqing Oilfield, northeastern China. Phylogenetic analysis based on the nearly complete 16S rRNA gene sequences revealed that strains DQW12E81-30(T) and DQW12E6-37-1 were members of family Rhodobacteraceae, which showed 95.6-95.9 % of 16S rRNA gene sequence similarities with Pararhodobacter aggregans DSM 18938(T), Rhodobacter veldkampii CGMCC 1.5006(T), and Roseinatronobacter thiooxidans DSM 13087(T), and lower similarities (<95.1 %) with all the left type species. Growth of strains DQW12E81-30(T) and DQW12E6-37-1 occurred at pH 7-8, 15-45 °C, and 0-4 % (w/v) of NaCl. The strains could grow both in dark and in light, but neither photosynthetic pigments nor photosynthetic reaction center gene pufM were detected in the strains. These photosynthesis-related features of the two isolates were different from those of Rhodobacter and Roseinatronobacter bacteria, but similar with those of Pararhodobacter. The genomic DNA G+C contents of strains DQW12E81-30(T) and DQW12E6-37-1 were 66.9 and 63.7 mol%, respectively. The predominant ubiquinone was Q-10 for both the strains. The major polar lipids of strain DQW12E81-30(T) were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, unidentified aminolipid, unidentified glycolipid, and unidentified phospholipid. The two strains had C18:1 ω7c, C18:0, and C18:1 ω7c 11-methyl as the major fatty acids. In addition, the strains DQW12E81-30(T) and DQW12E6-37-1 had C16:1 ω7c/C16:1 ω6c, C12:0, C14:0, C14:0 3-OH/C16:1 iso I, C10:0 3-OH, which were remarkably different from those of Pararhodobacter and Roseinatronobacter. The results of phenotypic, genotypic, and chemotaxonomic characteristics analyses indicated that strains DQW12E81-30(T) and DQW12E6-37-1 were readily different from their most phylogenetically closely related genera. Plastorhodobacter daqingensis gen. nov, sp. nov. is proposed for strains DQW12E81-30(T) and DQW12E6-37-1. The type strain is DQW12E81-30(T) (=LMG 27732(T)=CGMCC 1.12750(T)).

Glycocaulis albus sp. nov., a moderately halophilic dimorphic prosthecate bacterium isolated from petroleum-contaminated saline soil.

Two novel bacterial strains, SLG210-30A1(T) and SLG210-19A2, which shared 99.9 % 16S rRNA gene sequence similarity with each other, were isolated from petroleum-contaminated saline soil in Shengli Oilfield, eastern China. Cells were Gram-stain-negative, motile, aerobic, mesophilic and moderately halophilic. They could grow chemoheterotrophically with oxygen as an electron acceptor. Morphologically, cells were typical Caulobacteria-type dimorphic prosthecate bacteria. The genomic DNA G+C contents of strains SLG210-30A1(T) and SLG210-19A2 were 61.8 mol% and 61.6 mol% respectively. Strain SLG210-30A1(T) had Q10 as the predominant respiratory ubiquinone, and C16 : 0 (28.4 %), C17 : 0 (11.6 %), C18 : 0 (22.1 %) and C18 : 1ω7c (14.0 %) as the major cellular fatty acids. The polar lipids of the two isolates were some glycolipids, a lipid, a phospholipid, an aminoglycolipid and an aminophospholipid (all unidentified). The 16S rRNA gene sequences of strains SLG210-30A1(T) and SLG210-19A2 showed the highest similarities with Glycocaulis abyssi MCS 33(T) (99.8-99.9 %), but low sequence similarities (<94.7 %) with type strains of other members of the family Hyphomonadaceae. However, the DNA-DNA relatedness of G. abyssi MCS 33(T) to strains SLG210-30A1(T) and SLG210-19A2 was 37.4±4.4 % and 36.1±1.1 %, respectively. Based on different physiological, biochemical, and phylogenetic characteristics, strains SLG210-30A1(T) and SLG210-19A2 represent a novel species of the genus Glycocaulis. The name Glycocaulis albus is therefore proposed with strain SLG210-30A1(T) ( = LMG 27741(T) = CGMCC 1.12766(T)) as the type strain. An emended description of the genus Glycocaulis is also provided.

Ottowia shaoguanensis sp. nov., isolated from coking wastewater.

A Gram-negative, short rod-shaped, floc-forming bacterial strain J5-66(T) without any flagellum was isolated from coking wastewater collected from Shaoguan, Guangdong, China. It was capable of optimal growth at pH 7, 30 °C, and 1-2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belonged to the genus Ottowia in Comamonadaceae, and the highest 16S rRNA gene sequence similarity was 96.2 % with Ottowia pentelensis DSM 21699(T). The major cellular fatty acids of strain J5-66(T) were C16:1ω7c/C16:1 ω6c (45.0 %), C16:0 (21.1 %), C18:1 ω7c or/and C18:1 ω6c (19.2 %). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, glycolipid and two unidentified phospholipids (PL1 and PL2). The predominant ubiquinone was Q-8, and the G+C content of the genome DNA was 64.4 mol%. On the basis of genetic, phenotypic and chemotaxonomic analyses, strain J5-66(T) represents a novel species of the genus Ottowia for which the name Ottowia shaoguanensis sp. nov. is proposed. The type strain is J5-66(T) (=CGMCC 1.12431(T) =LMG 27408(T)).

Seohaeicola nanhaiensis sp. nov., a moderately halophilic bacterium isolated from the benthic sediment of South China Sea.

An aerobic, Gram-staining negative, non-motile, and rod-shaped bacterial strain, SS011A0-7#2-2(T), was isolated from the sediment of South China Sea with the depth of 1,500 m. Optimum growth occurred at pH 8.0, 30 °C, and 6 % (w/v) NaCl. Strain SS011A0-7#2-2(T) did not synthesize bacteriochlorophyll a or carotenoid, neither possess photosynthesis genes. Its genome DNA G+C content was 67.9 mol%. It contained Q-10 as the predominant ubiquinone and C18:1 ω7c (52.3 %) as the major fatty acid. The major polar lipids were phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, unidentified phospholipid, and unidentified aminolipid. The 16S rRNA gene sequence analysis revealed that it was closely related to Seohaeicola saemankumensis SD-15(T), Phaeobacter gallaeciensis BS 107(T) and Roseovarius pacificus 81-2(T) in Rhodobacteraceae, with the 16S rRNA gene sequence similarities being 96.5, 95.7, and 95.6 %, respectively. However, the phylogeny of the 16S rRNA gene sequences revealed that strain SS011A0-7#2-2(T) was a member of the genus Seohaeicola. Strain SS011A0-7#2-2(T) was moderately halophilic which was different from Seohaeicola saemankumensis SD-15(T), and it showed the enzyme activities and carbon source spectrum significantly different from Seohaeicola saemankumensis SD-15(T). As its physiological and chemotaxinomic properties were different from those of Seohaeicola saemankumensis SD-15(T), strain SS011A0-7#2-2(T) represents a novel species of the genus Seohaecola. The name Seohaeicola nanhaiensis sp. nov. is proposed, with strain SS011A0-7#2-2(T) (=LMG 27733(T) = CGMCC 1.12759(T)) as the type strain.

Nitratireductor shengliensis sp. nov., isolated from an oil-polluted saline soil.

Two Gram-negative, non-motile, short-rod-shaped bacterial isolates, designated 110399(T) and 110248, were isolated from an oil-polluted saline soil in Shengli Oilfield, Eastern China. The two strains shared 99.9 % 16S rRNA gene sequence similarity with the DNA-DNA relatedness value being 80.0 %. They were both capable to grow at 20-40 °C, pH 7-9, and 1-9 % (w/v) NaCl with the optimum growth happened at 30 °C, pH 8, and 2-6 % (w/v) NaCl. The phylogenetic analysis based on 16S rRNA gene sequences revealed that the two strains were members of Nitratireductor and most closely related to Nitratireductor pacificus pht-3B(T) and N. basaltis J3(T) with the 16S rRNA gene sequence similarities being 97.1 and 97.0 %. The DNA-DNA relatedness between the novel strains and two type strains were below 27 ± 7 %. The strains 110399(T) and 110248 also differed from N. pacificus and N. basaltis in nitrate reduction, salt tolerance, enzyme activities, and utilization of carbon sources. The major cellular fatty acids of strain 110399(T) were C19:0ω8c cyclo (10.5 %) and Summed Feature 8 (C18:1ω7c and/or C18:1ω6c, 41.5 %) which are typical in the genus Nitratireductor. The predominant ubiquinone was Q-10. The genome DNA G+C content of strain 110399(T) and 110248 was 61.1 and 61.7 mol%. On the basis of genetic, phenotypic, and chemotaxonomic analyses, strains 110399(T) and 110248 represent a novel species within the genus Nitratireductor, for which the name Nitratireductor shengliensis sp. nov. is proposed. The type strain is 110399(T) (=CGMCC 1.12519(T) = LMG 27405(T)).

[The synergism and mechanism of action of rClone30-hDR5 in combination with TRAIL on HCC].

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.

Isopropyl 3-phenyl-isoxazole-5-carboxyl-ate.

In the title compound, C13H13NO3, the isoxazole ring is approximately coplanar with the phenyl ring, the dihedral angle between their planes being 7.37 (19)°. In the crystal, centrosymmetrically related mol-ecules are linked into dimers by pairs of C-H⋯O hydrogen bonds, generating a ring of graph-set motif R 2 (2)(10).

Ethyl 4-benzamido-5-phenyl-4H-1,2,4-triazole-3-carboxyl-ate monohydrate.

In the title compound, C(18)H(16)N(4)O(3)·H(2)O, the dihedral angles between the triazole ring and the phenyl rings are 84.8 (4) and and 39.8 (4)°. The phenyl rings make a dihedral angle of 84.5 (9)°. In the crystal, the molecules are linked by N-H⋯O and O-H⋯N hydrogen bonds. An intra-molecular O⋯N inter-action also occurs [2.827 (3) Å].