Circulating miR-423-5p levels are associated with carotid atherosclerosis in patients with chronic kidney disease.

BACKGROUND AND AIMS: Carotid atherosclerosis is associated with an elevated risk of stroke in patients with chronic kidney disease. However, the molecular basis for the incidence of carotid atherosclerosis in patients with CKD is poorly understood. Here, we investigated whether circulating miR-423-5p is a crucial link between CKD and carotid atherosclerosis. METHODS AND RESULTS: We recruited 375 participants for a cross-sectional study to examine the occurrence of carotid plaque and plaque thicknesses. Levels of miR-423-5p were determined by qPCR analysis. We found that non-dialysis CKD patients had higher circulating exosomal and plasma miR-423-5p levels, and dialysis-dependent patients had lower miR-423-5p levels than non-dialysis CKD patients. After excluding for the influence of dialysis patients, linear regression analysis indicated that levels of circulating miR-423-5p are negatively correlated with eGFR (P < 0.001). Higher plasma miR-423-5p levels were associated with the incidence and severity of carotid plaques. In parallel, we constructed a murine model of CKD with a 5/6 nephrectomy protocol and performed RNA sequencing studies of aortic tissues. Consistent with these findings in CKD patients, circulating exosomal miR-423-5p levels in CKD mice were elevated. Furthermore, our RNA-seq studies indicated that the putative target genes of miR-423-5p were related to oxidative stress functions for aorta of CKD mice. CONCLUSION: Levels of miR-423-5p are associated with the presence and severity of carotid plaque in CKD. Data from our mouse model suggests that miR-423-5p likely influences gene expression programs related to oxidative stress in aorta of CKD mice.

Risk factors for foot ulcer recurrence in patients with comorbid diabetic foot osteomyelitis and diabetic nephropathy: A 3-year follow-up study.

This study aimed to explore the risk factors for foot ulcer recurrence in patients with comorbid diabetic foot osteomyelitis (DFO) and diabetic nephropathy (DN). This is a prospective cohort study. Between May 2018 and May 2021, we selected 120 inpatients with comorbid severe diabetic foot infection (PEDIS Grade 3 or above) and DN for inclusion in our study. All cases were followed up for 36 months. The study outcomes were whether foot ulcer recurred and the time to recurrence. The risk factors of ulcer recurrence were analysed by comparing the data of the three groups. According to the recurrence of foot ulcer, the participants were divided into three groups: Group A (no foot ulcer recurrence, n = 89), Group B (foot ulcer recurrence within 12-36 months, n = 19) and Group C (foot ulcer recurrence within 6-12 months, n = 12). The multivariate Cox regression analysis showed that urine albumin-creatinine ratio (UACR) (HR: 1.008, 95% CI: 1.005-1.011, P < .001) and vibration perception threshold (VPT) (HR: 1.064, 95% CI: 1.032-1.096, P < .001) were identified as independent risk factors. Kaplan-Meier curves showed a significant positive association between UACR or VPT and the risk of foot ulcer recurrence (log rank, all P < .05). Areas under the ROC curves for UACR, VPT and the combination of UACR and VPT were 0.802, 0.799 and 0.842, respectively. The best cut-off values of UACR and VPT were 281.51 mg/g and 25.12 V, respectively. In summary, elevated UACR and VPT were independent risk factors. The best clinical cut-off values of UACR and VPT for prediction of foot ulcer recurrence were 281.51 mg/g and 25.12 V, respectively. Besides, our results suggested that microcirculation disorders rather than macrovascular complications play a major role in the recurrence of foot ulcer in patients with comorbid DFO and DN.

The Clinical and Angiographic Outcomes of Postdilation after Percutaneous Coronary Intervention in Patients with Acute Coronary Syndrome: A Systematic Review and Meta-Analysis.

OBJECTIVE: The effect of postdilation in patients with acute coronary syndrome is still controversial. This meta-analysis aims to analyze the clinical and angiographic outcomes of postdilation after percutaneous coronary intervention in patients with acute coronary syndrome. METHODS: PubMed, Embase, the Cochrane Library, Web of Science, CNKI, and Wangfang databases were searched from inception to August 30, 2020. Eligible studies from acute coronary syndrome patients treated with postdilation were included. The primary clinical outcome was major adverse cardiovascular events (MACE), the secondary clinical outcomes comprised all-cause death, stent thrombosis, myocardial infarction, and target vessel revascularization, and the angiographic outcomes were no reflow and slow reflow. RESULTS: 11 studies met inclusion criteria. In clinical outcomes, our pooled analysis demonstrated that the postdilation had a tendency of decreasing MACE (OR = 0.67, 95% CI 0.45-1.00; P = 0.05) but significantly increased all-cause death (OR = 1.49, 95% CI 1.05-2.12; P = 0.03). No significant difference existed in stent thrombosis (OR = 0.71, 95% CI 0.40-1.26; P = 0.24), myocardial infarction (OR = 1.40, 95% CI 0.51-3.83; P = 0.51), and target vessel revascularization (OR = 0.61, 95% CI 0.21-1.80; P = 0.37) between postdilation and non-postdilation groups. In angiographic outcomes, there were no significant differences in no reflow (OR = 1.19, 95% CI 0.54-2.65; P = 0.66) and slow reflow (OR = 1.12, 95% CI 0.93-1.35; P = 0.24) between two groups. CONCLUSIONS: The postdilation tends to reduce the risk of MACE but significantly increases all-cause death, without significantly affecting stent thrombosis, myocardial infarction, target vessel revascularization, and coronary TIMI flow grade. However, more randomized controlled trials are required for investigating the effect of postdilation for patients with acute coronary syndrome (registered by PROSPERO, CRD42020160748).

Efficacy and safety of short-term 1-3 months versus standard 12 months dual antiplatelet therapy in patients undergoing percutaneous coronary intervention: a meta-analysis of randomized clinical trials.

Dual antiplatelet therapy (DAPT) is the basis of preventing stent thrombosis and ischemic events after percutaneous coronary intervention (PCI), but prolonging the duration of DAPT will increase the risk of bleeding. The optimal duration of DAPT after PCI remains controversial at present. The purpose of this meta-analysis was to investigate the efficacy and safety of short-term DAPT in patients undergoing PCI. PubMed, Embase, Cochrane and Web of science from inception to September 2019 were systematically searched. Randomized controlled trials were included to compare short term (3 months or less) with a standard 12-months DAPT in patients undergoing PCI. Random effect model and fixed effect model wereused to calculate the risk ratio (RR) and 95% confidence interval (CI) of each endpoint. This meta-analysis included 38479 patients undergoing PCI from 8 randomized clinical trials. No difference was observed in the risk of all-cause death (RR 0.92, 95% CI 0.80-1.06, P = 0.25), cardiovascular death (RR 0.88, 0.69-1.12, P = 0.29), myocardial infarction (RR 1.05, 0.94-1.19, P = 0.38), definite or probable stent thrombosis (RR 1.05, 0.80-1.36, P = 0.73), and stroke (RR 1.02, 0.80-1.30, P = 0.89) between short term and standard DAPT. The short-term DAPT could reduce the risk of major bleeding (RR 0.67, 0.48-0.94, P = 0.02) and any bleeding (RR 0.63, 0.48-0.82, P = 0.0005) compared with 12 months of DAPT. In conclusion, the short-term DAPT can reduce the risk of bleeding compared with standard DAPT, without increasing the risk of death or ischemia (Registered by PROSPERO, CRD42020153881).

Biochemical characterization of a bifunctional chitinase/lysozyme from Streptomyces sampsonii suitable for N-acetyl chitobiose production.

OBJECTIVES: Chitinases play important role in chitin bioconversion, while few of them have been put into use due to their poor properties. We aimed to identify and characterize chitinases suitable for N-acetyl chitooligosaccharides (COSs) production from chitin materials. RESULTS: A chitinase gene (SsChi28) from Streptomyces sampsonii XY2-7 was cloned and heterologously expressed in E. coli BL21 (DE3) as an active protein. The deduced protein shared high sequence identities and structure similarities with some glycoside hydrolase family 19 chitinases. The recombinant enzyme (SsChi28) was purified and biochemically characterized. SsChi28 was a monomeric protein with a molecular mass of 30 kDa estimated by SDS-PAGE. It was most active at pH 6.0 and 55 °C, respectively, and stable in a wide pH range of 3.5-11.5 and up to 60 °C. The enzyme exhibited strict substrate specificities towards ethylene glycol chitin (222.3 U/mg) and colloidal chitin (20.1 U/mg). Besides, it displayed lysozyme activity against Micrococcus lysodeikticus. SsChi28 hydrolyzed colloidal chitin to yield mainly N-acetyl chitobiose, accounting high up to 73% (w/w) in total products. CONCLUSION: The excellent enzymatic properties of SsChi28 may make it potential in chitin bioconversion (especially for N-acetyl COS production), as well as in biological control of fungal diseases.

Halorussus ruber sp. nov., isolated from an inland salt lake of China.

Halophilic archaeal strain YC25(T) was isolated from Yuncheng salt lake in Shanxi, China. Cells of strain YC25(T) were observed to be pleomorphic rods, stained Gram-negative, and formed red-pigmented colonies on solid media. Strain YC25(T) was found to be able to grow at 25-50 °C (optimum 37 °C), at 1.4-4.8 M NaCl (optimum 1.7 M), at 0-1.0 M MgCl2 (optimum 0.01 M), and at pH 5.5-9.0 (optimum pH 6.5). The cells lysed in distilled water, and the minimal NaCl concentration to prevent cell lysis was found to be 8 % (w/v). The major polar lipids of the strain were phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS), sulfated galactosyl mannosyl glucosyl diether (S-TGD-1), sulfated mannosyl glucosyl diether (S-DGD-1), galactosyl mannosyl glucosyl diether (TGD-1), mannosyl glucosyl diether (DGD-1), and an unknown diglycosyl diether (DGD-2) chromatographically identical to those of Halorussus rarus CGMCC 1.10122(T). The 16S rRNA gene and rpoB' gene of strain YC25(T) were phylogenetically related to the corresponding genes of Halorussus rarus CGMCC 1.10122(T) (94.3-95.4 and 91.5 % nucleotide identity, respectively). The DNA G+C content of strain YC25(T) was determined to be 63.3 mol%. The phenotypic, chemotaxonomic, and phylogenetic properties suggested that strain YC25(T) (=CGMCC 1.12122(T) = JCM 18363(T)) represents a new species of Halorussus, for which the name Halorussus ruber sp. nov. is proposed.

Haloarchaeobius amylolyticus sp. nov., isolated from a marine solar saltern.

Halophilic archaeal strain XD48(T) was isolated from a Chinese marine solar saltern. Cells were pleomorphic, stained Gram-negative and formed red-pigmented colonies on solid media. Strain XD48(T) was found to be able to grow at 25-50 °C (optimum 37 °C), at 0.9-4.8 M NaCl (optimum 3.1 M NaCl), at 0-1.0 M MgCl2 (optimum 0.3 MgCl2) and at pH 5.0-9.5 (optimum pH 6.5). The cells lysed in distilled water, and the minimal NaCl concentration to prevent cell lysis was found to be 5% (w/v). The major polar lipids of the strain were phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS), sulfated galactosyl mannosyl glucosyl diether (S-TGD-1), sulfated mannosyl glucosyl diether (S-DGD-1) and six unknown glycolipids. The 16S rRNA gene and rpoB' gene of strain XD48(T) were phylogenetically related to the corresponding genes of Haloarchaeobius members (92.4-93.9 and 89.6-90.5% similarities, respectively). The DNA G + C content of strain XD48(T) was determined to be 65.3 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain XD48(T) (=CGMCC 1.12230(T) = JCM 18642(T)) represents a new species of Haloarchaeobius, for which the name Haloarchaeobius amylolyticus sp. nov. is proposed.

Halomicroarcula salina sp. nov., isolated from a marine solar saltern.

Halophilic archaeal strain YGHS18(T) was isolated from the Yinggehai marine solar saltern near Shanya city of Hainan Province, China. Cells from the strain were observed to be pleomorphic rods, stained Gram-negative, and formed red-pigmented colonies on solid media. Strain YGHS18(T) was found to be able to grow at 20-50 °C (optimum 37 °C), with 0.9-4.8 M NaCl (optimum 2.1 M) and at pH 5.5-9.0 (optimum pH 7.0). The cells lysed in distilled water and the minimum NaCl concentration to prevent cell lysis was found to be 0.9 M. The major polar lipids of the strain were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, glucosyl mannosyl glucosyl diether and a diglycosyl diether (DGD-2). Strain YGHS18(T) possessed two heterogeneous 16S rRNA genes (rrnA and rrnB) and both were related to those of members of the genera Haloarcula (93.1-96.9% sequence similarity) and Halomicroarcula (92.7-96.1% similarity). The rrnA gene (orthologous gene) of strain YGHS18(T) clustered phylogenetically with members of the genus Halomicroarcula while the rrnB gene formed a paraphyly with members of the genera Halomicroarcula and Haloarcula . The rpoB' gene of strain YGHS18(T) was related phylogenetically to species of the genera Halomicroarcula (91.6-92.7% sequence similarity) and Haloarcula (91.5-92.4% similarity). EF-2 gene analysis revealed that strain YGHS18(T) was related phylogenetically to species of the genus Halomicroarcula (92.2-92.9% sequence similarity) rather than to those of the genus Haloarcula (90.9-91.7% similarity). The DNA G+C content of strain YGHS18(T) was determined to be 64.5 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YGHS18(T) ( =CGMCC 1.12128(T) =JCM 18369(T)) represents a novel species of the genus Halomicroarcula , for which the name Halomicroarcula salina sp. nov. is proposed.

Haloarchaeobius salinus sp. nov., isolated from an inland salt lake, and emended description of the genus Haloarchaeobius.

The halophilic archaeal strain, YC82(T), was isolated from Yuncheng salt lake in Shanxi, PR China. Cells from strain YC82(T) were Gram-stain negative, pleomorphic rods, which lysed in distilled water and formed light-red colonies on solid media. Strain YC82(T) grew at 25-50 °C (optimum 37 °C), in 1.4-4.8 M NaCl (optimum 2.0 M), in 0-1.0 M MgCl2 (optimum 0.05 M) and at pH 6.0-9.5 (optimum pH 7.5). The major polar lipids of strain YC82(T) were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and three glycolipids, which were chromatographically identical to those of Haloarchaeobius iranensis IBRC-M 10013(T) and Haloarchaeobius litoreus CGMCC 1.10390(T). 16S rRNA gene analysis revealed that strain YC82(T) had two dissimilar 16S rRNA genes and that it was phylogenetically related to Hab. iranensis IBRC-M 10013(T) (94.3-99.0 % nucleotide identity) and Hab. litoreus CGMCC 1.10390(T) (94.1-98.8 % nucleotide identity). The rpoB' gene similarities between strain YC82(T) and Hab. iranensis IBRC-M 10013(T) and Hab. litoreus CGMCC 1.10390(T) were 96.5 % and 95.7 %, respectively. The DNA G+C content of strain YC82(T) was 63.7 mol%. Strain YC82(T) showed low DNA-DNA relatedness with Hab. iranensis IBRC-M 10013(T) and Hab. litoreus CGMCC 1.10390(T). The phenotypic, chemotaxonomic and phylogenetic properties of strain YC82(T) ( = CGMCC 1.12232(T) = JCM 18644(T)) suggest that it represents a novel species of the genus Haloarchaeobius, for which the name Haloarchaeobius salinus sp. nov. is proposed. An emended description of the genus Haloarchaeobius is also presented.

Halorussus amylolyticus sp. nov., isolated from an inland salt lake.

A halophilic archaeal strain, YC93T, was isolated from Yuncheng salt lake in Shanxi Province, China. Cells were pleomorphic rods, stained Gram-negative and formed light-red-pigmented colonies on agar plates. Strain YC93T was able to grow at 25­50 °C (optimum 37 °C), with 1.4­4.8 M NaCl (optimum 2.0 M), with 0­1.0 M MgCl2 (optimum 0.05 M) and at pH 6.0­9.5 (optimum pH 7.0). Cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 8 % (w/v). 16S rRNA gene sequence analysis revealed that strain YC93T had two dissimilar 16S rRNA genes both of which were phylogenetically related to those of the two recognized members of the genus Halorussus (93.0­95.3 % similarity). The rpoB' gene of strain YC93T was phylogenetically related to the corresponding gene of Halorussus rarus TBN4T (91.3 % similarity) and Halorussus ruber YC25T (90.5 %). The major polar lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and five glycolipids chromatographically identical to those of Halorussus rarus CGMCC 1.10122T. The DNA G+C content of strain YC93T was 64.6 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YC93T represents a novel species of the genus Halorussus, for which the name Halorussus amylolyticus sp. nov. is proposed. The type strain is YC93T ( = CGMCC 1.12126T = JCM 18367T).

Halosimplex litoreum sp. nov., isolated from a marine solar saltern.

A halophilic archaeal strain, YGH94(T), was isolated from the Yinggehai marine solar saltern near the Shanya city of Hainan Province, China. Cells of the strain were observed to be short rods, stain Gram-negative and to form red-pigmented colonies on solid media. Strain YGH94(T) was found to grow at 25-50 °C (optimum 40 °C), at 0.9-4.8 M NaCl (optimum 3.1 M), at 0-1.0 M MgCl2 (optimum 0.05 M) and at pH 5.0-9.0 (optimum pH 7.5). The cells were found to lyse in distilled water and the minimal NaCl concentration to prevent cell-lysis was determined to be 5 % (w/v). The major polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and four major glycolipids (disulfated mannosyl glucosyl diether, sulfated mannosyl glucosyl diether and two unidentified glycolipids chromatographically identical to glycolipids in Halosimplex carlsbadense JCM 11222(T)). Strain YGH94(T) was found to possess two heterogeneous 16S rRNA genes (rrnA and rrnB) and both are related to those of Hsx. carlsbadense JCM 11222(T) (92.7-98.6 % similarities), Halosimplex pelagicum R2(T) (94.6-99.2 % similarities) and Halosimplex rubrum R27(T) (92.9-98.8 % similarities). The rpoB' gene similarity between strain YGH94(T) and Hsx. carlsbadense JCM 11222(T), Hsx. pelagicum R2(T) and Hsx. rubrum R27(T) are 95.4, 94.9 and 95.1 %, respectively. The DNA G+C content of strain YGH94(T) was determined to be 64.0 mol%. Strain YGH94(T) showed low DNA-DNA relatedness (35-39 %) with the current three members of the genus Halosimplex. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain YGH94(T) (=CGMCC 1.12235(T) = JCM 18647(T)) represents a new species of the genus Halosimplex, for which the name Halosimplex litoreum sp. nov. is proposed.

Halovenus rubra sp. nov., isolated from salted brown alga Laminaria.

Halophilic archaeal strain R28(T) was isolated from the brown alga Laminaria produced at Dalian, Liaoning Province, China. The cells of the strain were pleomorphic and lysed in distilled water, stained Gram-negative, and formed red-pigmented colonies. Strain R28(T) was able to grow at 25-50 °C (optimum 42 °C), in the presence of 3.1-5.1 M NaCl (optimum 3.9 M NaCl), with 0.005-1.0 M MgCl(2) (optimum 0.01 M MgCl(2)) and at pH 6.0-9.5 (optimum pH 7.0-7.5). The minimal NaCl concentration to prevent cell lysis was 15 % (w/v). The major polar lipids of the strain were identified as phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, and two glycolipids chromatographically identical to those of Halovenus aranensis CGMCC 1.11001(T). The 16S rRNA gene and rpoB' gene of strain R28(T) were phylogenetically related to the corresponding genes of Hvn. aranensis CGMCC 1.11001(T) (91.9-97.2 and 82.9 % nucleotide identity, respectively). The DNA G+C content of strain R28(T) was determined to be 56.3 mol%. The phenotypic, chemotaxonomic, and phylogenetic properties suggest that strain R28(T) (=CGMCC 1.10592(T) = JCM 17269(T)) represents a novel species of the genus Halovenus, for which the name Halovenus rubra sp. nov. is proposed.

Halorientalis brevis sp. nov., Isolated from an Inland Salt Lake of China.

Halophilic archaeal strain YC89(T) was isolated from Yuncheng salt lake in Shanxi, China. Cells from strain YC89(T) were short rods, lysed in distilled water, stained Gram-negative and formed red-pigmented colonies on agar plate. Strain YC89(T) was able to grow at 25-50°C (optimum 37°C), at 1.4-4.8 M NaCl (optimum 2.6-3.1 M), at 0-1.0 M MgCl2 (optimum 0.3 M) and at pH 6.0-9.5 (optimum pH 7.5). The major polar lipids are phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, sulfated mannosyl glucosyl diether and two unknown glycolipids. 16S rRNA gene analysis revealed that strain YC89(T) was phylogenetically related to Halorientalis persicus D108(T) (95.6% nucleotide identity) and H. regularis TNN28(T) (95.3% nucleotide identity). The rpoB' gene similarities between strain YC89(T) and H. persicus IBRC-M 10043(T) and H. regularis TNN28(T) were 88.1 and 88.0%, respectively. The DNA G+C content of strain YC89(T) was determined to be 61.3 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YC89(T) (=CGMCC 1.12125(T) = JCM 18366(T)) represents a new species of Halorientalis, for which the name H. brevis sp. nov. is proposed.

Halorubrum salinum sp. nov., isolated from a marine solar saltern.

The halophilic archaeal strain GX71(T) was isolated from the Gangxi marine solar saltern near the Weihai city of Shandong Province, China. Cells of the strain were pleomorphic and lysed in distilled water, stained Gram-negative and formed red-pigmented colonies. Strain GX71(T) was able to grow at 25-45 °C (optimum 30 °C), in the presence of 1.7-4.8 M NaCl (optimum 2.6 M NaCl), with 0.005-0.7 M MgCl2 (optimum 0.05 M MgCl2) and at pH 5.5-9.5 (optimum pH 7.0-7.5). Cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 10 % (w/v). The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, one major glycolipid chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-3) and an unidentified lipid was also detected. The 16S rRNA gene sequence of strain GX71(T) showed 94.0-97.0 % similarity to members of the genus Halorubrum of the family Halobacteriaceae. The rpoB' gene sequence of strain GX71(T) was 87.3-93.4 % similarity to current members of the genus Halorubrum. The DNA G+C content of GX71(T) was 67.1 mol%. Strain GX71(T) showed low DNA-DNA relatedness with Halorubrum lipolyticum CGMCC 1.5332(T), Halorubrum saccharovorum CGMCC 1.2147(T), Halorubrum kocurii CGMCC 1.7018(T) and Halorubrum arcis CGMCC 1.5343(T), the most closely related members of the genus Halorubrum. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX71(T) represents a novel species of the genus Halorubrum, for which the name Halorubrum salinum sp. nov. is proposed. The type strain is GX71(T) (= CGMCC 1.10458(T) = JCM 17093(T)).

Salinigranum rubrum gen. nov., sp. nov., a member of the family Halobacteriaceae isolated from a marine solar saltern.

Halophilic archaeal strain GX10(T) was isolated from the Gangxi marine solar saltern in China. Strain GX10(T) was observed to have pleomorphic cells that lysed in distilled water, stained Gram-negative and produced red-pigmented colonies. Strain GX10(T) was able to grow at 20-50 °C (optimum 37 °C), with 1.4-4.8 M NaCl (optimum 3.1 M NaCl), with 0-0.7 M MgCl2 (optimum 0.05 M MgCl2) and at pH 5.0-9.0 (optimum pH 7.0). The major polar lipids of strain GX10(T) were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, two major glycolipids chromatographically identical to sulfated mannosyl glucosyl diether and mannosyl glucosyl diether, and five unidentified glycolipids. Phylogenetic tree reconstructions based on 16S rRNA gene and rpoB' sequences revealed that strain GX10(T) was distinct from the related genera, Halogranum, Haloferax, Halopelagius, Halogeometricum, Halobellus, Haloplanus and Halorubrum. The DNA G+C content of strain GX10(T) was 62.9 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain GX10(T) represents a novel species of a new genus within the family Halobacteriaceae, for which the name Salinigranum rubrum gen. nov., sp. nov. is proposed. The type strain of the type species is GX10(T) ( = CGMCC 1.10385(T) = JCM 17116(T)).

Halomicroarcula limicola sp. nov., isolated from a marine solar saltern, and emended description of the genus Halomicroarcula.

Halophilic archaeal strain YGHS32T was isolated from the Yinggehai marine solar saltern near Shanya city of Hainan Province, China. Cells of the strain were pleomorphic and lysed in distilled water, stained Gram-negative and formed red-pigmented colonies. Strain YGHS32T was able to grow at 20-50 °C (optimum 37 °C), in the presence of 0.9-4.8 M NaCl (optimum 2.1 M NaCl), with 0.005-1.0 M MgCl2 (optimum 0.3 M MgCl2) and at pH 6.0-8.5 (optimum pH 7.5). The minimal NaCl concentration to prevent cell lysis was 5% (w/v). The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and four major glycolipids chromatographically identical to sulfated mannosyl glucosyl diether, mannosyl glucosyl diether, glucosyl mannosyl glucosyl diether and a diglycosyl diether. Strain YGHS32T had two dissimilar 16S rRNA genes and both of them were phylogenetically related to those of Halomicroarcula pellucida JCM 17820T (92.9-96.3% sequence similarity). The rpoB' gene sequence similarity between strain YGHS32T and Halomicroarcula pellucida JCM 17820T was 91.3%. The DNA G+C content of strain YGHS32T was 64.0 mol%. The DNA-DNA hybridization value between strain YGHS32T and Halomicroarcula pellucida JCM 17820T was 45%. It was concluded that strain YGHS32T (=CGMCC 1.12129T=JCM 18640T) represents a novel species of the genus Halomicroarcula, for which the name Halomicroarcula limicola sp. nov. is proposed. An emended description of the genus Halomicroarcula is also presented.

Haloarchaeobius litoreus sp. nov., isolated from a marine solar saltern.

Two extremely halophilic archaeal strains GX1(T) and GX60 were isolated from the Gangxi marine solar saltern, China. Cells from the two strains were observed to be rod-shaped and stained Gram-negative, with red-pigmented colonies. Strains GX1(T) and GX60 were found to be able to grow at 25-50 °C (optimum 37 °C), at 1.4-4.8 M NaCl (optimum 2.6 M), at pH 5.5-9.5 (optimum pH 7.0) and neither strain required Mg(2+) for growth. The cells lysed in distilled water and the minimal NaCl concentration to prevent cell-lysis was found to be 8 % (w/v). The major polar lipids of the two strains were identified as phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and three glycolipids chromatographically identical to those of Haloarchaeobius iranensis IBRC-M 10013(T). 16S rRNA gene analysis revealed that each strain had two dissimilar 16S rRNA genes and both strains were phylogenetically related to Hab. iranensis IBRC-M 10013(T) (94.9-98.9 % nucleotide identity). The rpoB' gene similarity between strains GX1(T) and GX60, and between these strains and Hab. iranensis IBRC-M 10013(T) were found to be 99.6, 96.0 and 95.8 %, respectively. The DNA G + C content of strain GX1(T) and GX60 were determined to be 67.7 and 67.8 mol %, respectively. The DNA-DNA hybridization value of strains GX1(T) and GX60 was 86 % and the two strains showed low DNA-DNA relatedness with Hab. iranensis IBRC-M 10013(T) (38 and 32 %). It was concluded that strain GX1(T) (= CGMCC 1.10390(T) = JCM 17114(T)) and strain GX60 (= CGMCC 1.10389 = JCM 17120) represent a new species of Haloarchaeobius, for which the name Haloarchaeobius litoreus sp. nov. is proposed.

Halobellus litoreus sp. nov., a halophilic archaeon isolated from a Chinese marine solar saltern.

Halophilic archaeal strain GX31(T) was isolated from a marine solar saltern of China. The cells of the strain were rod-shaped and lysed in distilled water, stain Gram-negative and formed red-pigmented colonies. It was neutrophilic, and required at least 0.9 M NaCl and 0-1.0 M MgCl2 for growth under the optimum growth temperature of 37 °C. The major polar lipids of the strain were phosphatidylglycerol (PG), PG phosphate methyl ester, PG sulphate, and two major glycolipids chromatographically identical to sulphated mannosyl glucosyl diether (S-DGD-1) and mannosyl glucosyl diether (DGD-1), respectively. Trace amounts of two unidentified lipids were also detected. On the basis of 16S rRNA gene sequence analysis, strain GX31(T) was closely related to the members of Halobellus of the family Halobacteriaceae with similarities of 94.1-98.7 %. Strain GX31(T) showed 89.8-95.4 % of the rpoB' gene similarity to the members of Halobellus. The DNA G+C content of strain GX31(T) was 66.8 mol%. Strain GX31(T) showed low DNA-DNA relatedness with two most related members of the genus Halobellus. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX31(T) represent a novel species of the genus Halobellus, for which the name Halobellus litoreus sp. nov. is proposed. The type strain is GX31(T) (=CGMCC 1.10387(T) = JCM 17118(T)).

Catalytic hairpin assembly-based AIEgen/graphene oxide nanocomposite for fluorescence-enhanced and high-precision spatiotemporal imaging of microRNA in living cells.

The low abundance, heterogeneous expression, and temporal changes of miRNA in different cellular locations pose significant challenges for both the detection sensitivity of miRNA liquid biopsy and intracellular imaging. In this work, we report an intelligently assembled biosensor based on catalytic hairpin assembly (CHA) and aggregation-induced emission (AIE), named as catalytic hairpin aggregation-induced emission (CHAIE), for the ultrasensitive detection and intracellular imaging of miRNA-155. To achieve such goal, tetraphenylethylene-N3 (TPE-N3) is used as AIE luminogen (AIEgen), while graphene oxide is introduced to quench the fluorescence. When the target miRNA is present, CHA reaction is triggered, causing the AIEgen to self-assemble with the hairpin DNA. This will restrict the intramolecular rotation of the AIEgen and produce a strong AIE fluorescence. Interestingly, CHAIE does not require any enzyme or expensive thermal cycling equipment, and therefore provides a rapid detection. Under optimal conditions, the proposed biosensor can determine miRNA in the concentration range from 2 pM to 200 nM within 30 min, with the detection limit of 0.42 pM. The proposed CHAIE biosensor in this work offers a low background signal and high sensitivity, making it applicable for highly precise spatiotemporal imaging of target miRNA in living cells.

The putative auxin efflux carrier OsPIN3t is involved in the drought stress response and drought tolerance.

The phytohormone auxin plays a critical role in plant growth and development, and its spatial distribution largely depends on the polar localization of the PIN-FORMED (PIN) auxin efflux carrier family members. In this study, we identify a putative auxin efflux carrier gene in rice, OsPIN3t, which acts in auxin polar transport but is also involved in the drought stress response in rice. We show that OsPIN3t-GFP fusion proteins are localized in plasma membranes, and this subcellular localization changes under 1-N-naphthylphthalamic acid (NPA) treatment. The tissue-specific expression patterns of OsPIN3t were also investigated using a ß-glucuronidase (GUS) reporter, which showed that OsPIN3t was mainly expressed in vascular tissue. The GUS activity in OsPIN3tpro::GUS plants increased by NAA treatment and decreased by NPA treatment. Moreover, knockdown of OsPIN3t caused crown root abnormalities in the seedling stage that could be phenocopied by treatment of wild-type plants with NPA, which indicated that OsPIN3t is involved in the control of polar auxin transport. Overexpression of OsPIN3t led to improved drought tolerance, and GUS activity significantly increased when OsPIN3tpro::GUS plants were subjected to 20% polyethylene glycol stress. Taken together, these results suggest that OsPIN3t is involved in auxin transport and the drought stress response, which suggests that a polar auxin transport pathway is involved in the regulation of the response to water stress in plants.