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Backgrounds The evaluation of liver fibrosis stages is essential for the clinical management of chronic hepatitis B (CHB). Aims To develop and validate a novel noninvasive index for moderate to severe fibrosis (≥S2) in CHB patients. Methods A total of 401 CHB patients who underwent liver biopsy were divided into the training (n = 300) and validation (n = 101) cohort. Histological severity was scored using a modified Scheuer system. Clinical and laboratory assessments were collected. Results In the training cohort, PACG, a novel index combining the quantitative hepatitis B core antibody (qAnti-HBc), platelet count (PLT), and albumin globulin ratio (A/G), presented better diagnostic performance (AUROC = 0.814) than that of APRI (0.735, p = 0.007) and FIB-4 (0.749, p = 0.014). In the validation cohort, the AUROC of the PACG, APRI, FIB-4 and Fibroscan were 0.834, 0.806, 0.791 and 0.810, respectively. More importantly, a higher and lower cutoff of PACG for predicting ≥S2 fibrosis or not had a >90% sensitivity and specificity, with a diagnostic accuracy of 85.9%. Conclusion PACG is a promising noninvasive alternative to liver biopsy in CHB patients for the evaluation of moderate to severe fibrosis.

To identify and characterize a novel aryloxyphenoxypropionate (AOPP) herbicide-hydrolyzing carboxylesterase from Aquamicrobium sp. FPB-1. A carboxylesterase gene, fpbH, was cloned from Aquamicrobium sp. FPB-1. The gene is 798 bp long and encodes a protein of 265 amino acids. FpbH is smaller than previously reported AOPP herbicide-hydrolyzing carboxylesterases and shares only 21–35% sequence identity with them. FpbH was expressed in Escherichia coli BL21(DE3) and the product was purified by Ni–NTA affinity chromatography. The purified FpbH hydrolyzed a wide range of AOPP herbicides with catalytic efficiency in the order: haloxyfop-P-methyl > diclofop-methyl > fenoxaprop-P-ethyl > quizalofop-P-ethyl > fluazifop-P-butyl > cyhalofop-butyl. The optimal temperature and pH for FpbH activity were 37 °C and 7, respectively. FpbH is a novel AOPP herbicide-hydrolyzing carboxylesterase; it is a good candidate for mechanistic study of AOPP herbicide-hydrolyzing carboxylesterases and for bioremediation of AOPP herbicide-contaminated environments.

The stable HBV-transfected cell lines, which based on stable integration of replication-competent HBV genome into hepatic cells, are widely used in basic research and antiviral drug evaluation against HBV. However, previous reported strategies to generate HBV-replicating cell lines, which primarily rely on random integration of exogenous DNA by plasmid transfection, are inefficient and time-consuming. We newly developed an all-in-one Sleeping Beauty transposon system (denoted pTSMP-HBV vector) for robust generation of stable HBV-replicating cell lines of different genotype. The pTSMP-HBV vector contains HBV 1.3-copy genome and dual selection markers (mCherry and puromycin resistance gene), allowing rapid enrichment of stably-transfected cells via red fluorescence-activated cell sorting and puromycin antibiotic selection. In this protocol, we described the detailed procedure for constructing the HBV-replicating stable cells and systematically evaluating HBV replication and viral protein expression profiles of these cells.

To investigate the effects of curcumin on pain threshold and the expressions of nuclear factor κ B (NF-κ B) and CX3C chemokine receptor 1 (CX3CR1) in spinal cord and dorsal root ganglion (DRG) of the rats with sciatic nerve chronic constrictive injury.One hundred and twenty male Sprague Dawley rats, weighing 220-250 g, were randomly divided into 4 groups. Sham surgery (sham) group: the sciatic nerves of rats were only made apart but not ligated; chronic constrictive injury (CCI) group: the sciatic nerves of rats were only ligated without any drug treatment; curcumin treated injury (Cur) model group: the rats were administrated with curcumin 100 mg/(kg·d) by intraperitoneal injection for 14 days after CCI; solvent control (SC) group: the rats were administrated with the solvent at the same dose for 14 days after CCI. Thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) of rats were respectively measured on pre-operative day 2 and postoperative day 1, 3, 5, 7, 10 and 14. The lumbar segment L4-5 of the spinal cord and the L4, L5 DRG was removed at post-operative day 3, 7 and 14. The change of nuclear factor κ B (NF-κ B) p65 expression was detected by Western blotting while the expression of CX3CR1 was determined by immunohistochemical staining.Compared with the sham group, the TWL and MWT of rats in the CCI group were significantly decreased on each post-operative day (P0.01), which reached a nadir on the 3rd day after CCI, and the expressions of NF-κ B p65 and CX3CR1 were markedly increased in spinal cord dorsal horn and DRG. In the Cur group, the TWL of rats were significantly increased than those in the CCI group on post-operative day 7, 10 and 14 (P0.05) and MWT increased than those in the CCI group on post-operative day 10 and 14 (P0.05). In addition, the administration of curcumin significantly decreased the positive expressions of NF-κ B p65 and CX3CR1 in spinal cord and DRG (P0.05).Our study suggests that curcumin could ameliorate the CCI-induced neuropathic pain, probably through inhibiting CX3CR1 expression by the activation of NF-κ B p65 in spinal cord and DRG.

A non-sporulating, non-motile, catalase- and oxidase-positive, Gram-negative, rod-shaped bacterial strain, designated BA-3T, was isolated from activated sludge of a wastewater treatment facility. The strain was able to degrade about 95 % of 100 mg 3-phenoxybenzoic acid l−1 within 2 days of incubation. Growth occurred in the presence of 0–2 % (w/v) NaCl [optimum, 0.5 % (w/v) NaCl], at pH 5.5–9.0 (optimum, pH 7.0) and at 10–37 °C (optimum, 28 °C). Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain BA-3T was a member of the genus Sphingobium ; it showed highest gene sequence similarity to Sphingobium qiguonii X23T (98.2 %), and similarities of Sphingobium species. The polar lipid pattern, the presence of spermidine and ubiquinone Q-10, the predominance of summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) in the cellular fatty acid profile and the DNA G+C content also supported affiliation of the isolate to the genus Sphingobium . Strain BA-3T showed low DNA–DNA relatedness values (21.3±0.8 %) with Sphingobium qiguonii X23T. Based on phenotypic, genotypic and phylogenetic data, strain BA-3T represents a novel species of the genus Sphingobium , for which the name Sphingobium jiangsuense sp. nov. is proposed; the type strain is BA-3T ( = CCTCC AB 2010217T = KCTC 23196T = KACC 16433T).

A novel facultatively anaerobic, non-spore-forming, non-motile, catalase- and oxidase-positive, Gram-negative and rod-shaped bacterial strain, designated Y12T, was isolated from activated sludge of a wastewater bio-treatment facility. The strain was able to degrade about 90 % of added propanil (100 mg l−1) within 3 days of incubation. Growth occurred in the presence of 0–4.5 % (w/v) NaCl (optimum 0.5 %), at 10–40 °C (optimum 28 °C) and at pH 5.5–10.0 (optimum pH 7.0). Vesicular internal membrane structures and photoheterotrophic growth were not observed. The major respiratory quinone was ubiquinone-10 and the major cellular fatty acid was summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The genomic DNA G+C content of strain Y12T was 63.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain Y12T was a member of the genus Catellibacterium, as it showed highest sequence similarities to Catellibacterium caeni DCA-1T (99.1 %) and Catellibacterium. Strain Y12T showed low DNA–DNA relatedness values with C. caeni DCA-1T. Based on phenotypic, genotypic and phylogenetic properties, strain Y12T represents a novel species of the genus Catellibacterium, for which the name Catellibacterium nanjingense sp. nov. is proposed. The type strain is Y12T ( = CCTCC AB 2010218T = KCTC 23298T). An emended description of the genus Catellibacterium is also presented.

A Gram-negative, non-motile, non-spore-forming, non-flagellated rod capable of degrading cypermethrin, designated LQY-18T, was isolated from activated sludge of a wastewater treatment plant in China. Strain LQY-18T grew at 8–40 °C (optimum 30 °C), at pH 5.0–10.0 (optimum pH 7.0) and with 0–5 % (w/v) NaCl (optimum 1 %). The predominant menaquinone was MK-7 (97 %) and the major fatty acids were summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c), iso-C15 : 0 and iso-C17 : 0 3-OH. The DNA G+C content was 40.3 mol%. Phylogenetic analysis revealed that the isolate belonged to the genus Sphingobacterium of the phylum Bacteroidetes and showed low 16S rRNA gene sequence similarity with recognized members of the genus Sphingobacterium. The closest neighbour was Sphingobacterium mizutaii ATCC 33299T (92.9 % 16S rRNA gene sequence similarity). On the basis of phenotypic, genetic and phylogenetic data, strain LQY-18T ( = ACCC 05410T = CCTCC AB 2010005T = KCTC 23009T) should be classified as a representative of a novel species of the genus Sphingobacterium, for which the name Sphingobacterium wenxiniae sp. nov. is proposed.

A synthetic pyrethroid (SP)-degrading bacterium, designated pyd-1, was isolated from SPcontaminated soil. Based on its phenotypic and genotypic properties, the strain was identified as Ochrobactrum tritici. Strain pyd-1 was able to degrade a wide range of SPs, and its degradation efficiencies were dependent on the molecular structure of the SP. Interestingly, the strain degraded cis- and trans-permethrin (cypermethrin) at nearly the same rate and possessed approximately equal hydrolysis activities toward the two enantiomers of fenpropathrin. These results suggest that different isomers of SPs are degraded with equal efficiency by strain pyd-1. We studied the metabolic pathway of fenpropathrin degradation in strain pyd-1 by metabolite identification and enzymatic analysis. Fenpropathrin is degraded by hydrolysis of the carboxylester linkage to yield 2,2,3,3-tetramethylcyclopropanecarboxylic acid and 3-phenoxybenzaldehyde, which is converted to 3-phenoxybenzoic acid (PBA). PBA is further metabolized to 4-hydroxy-3-phenoxybenzoic acid (4-hydroxy-PBA). 4-Hydroxy-PBA is oxidized to protocatechuate and p-hydroquinone. Protocatechuate is further oxidized through an ortho-cleavage pathway, and p-hydroquinone is degraded via 1,2,4-benzenetriol.

A bacterial strain, designated BF-3T, was isolated from phenol-contaminated soil and investigated using a polyphasic taxonomic approach. Cells were Gram-reaction-negative, non-sporulating, non-motile, short rods. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BF-3Tformed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genusComamonas; it showed highest sequence similarities toComamonas aquaticaLMG 2370T(96.8 %),C. nitrativoransDSM 13191T(96.4 %),C. odontotermitisLMG 23579T(96.4 %),C. kerstersiiLMG 3475T(96.3 %),C. koreensisKCTC 12005T(96.1 %) andC. terrigenaLMG 1253T(96.0 %). The major cellular fatty acids were C16 : 0, C18 : 1/C18 : 1ω7c, C17 : 0cyclo and summed feature 3 (C16 : 1ω7cand/or iso-C15 : 02-OH). Based on the phylogenetic analysis, DNA–DNA hybridization, whole-cell fatty acid composition and biochemical characteristics, strain BF-3Twas clearly distinct from type strains of other recognized species of the genusComamonasand, as such, represents a novel species of the genusComamonas, for which the nameComamonas zonglianiisp. nov. is proposed. The type strain is BF-3T(=CCTCC AB 209170T=DSM 22523T).

A bacterial strain, pcnb-21, capable of degrading pentachloronitrobenzene (PCNB) under aerobic and anoxic conditions, was isolated from a long-term PCNB-polluted soil by an enrichment culture technique and identified as Labrys portucalensis based upon its morphological, physiological and biochemical properties, as well as 16S rRNA gene sequence analysis. Effects of different factors, such as temperature and pH, on PCNB biodegradation were studied. Strain pcnb-21 efficiently degraded PCNB at temperatures from 20 to 30 °C and initial pH values from 4 to 7, which might be the first time that a Labrys strain was found capable of efficiently degrading PCNB. The degradation of PCNB was affected by oxygen, and the degradation decreased with increasing aeration. Exogenous electron donors such as glucose, lactic acid and succinic acid promoted the biodegradation of PCNB, while electron acceptors such as sodium nitrite, sodium sulfate, sodium nitrate and sodium sulfate inhibited PCNB biodegradation. The degradation of PCNB in sterile and non-sterile soils by a green fluorescent protein (GFP)-labeled strain, pcnb-21- gfp , was also studied. Cells of pcnb-21- gfp efficiently degraded 100 mg kg −1 PCNB in sterile and non-sterile soils and could not be detected after 42 days. Strain pcnb-21 might be useful in bioremediating PCNB-polluted soils and environment.

OBJECTIVE: To investigate the effects of curcumin on the behavior of chronic constrictive injury (CCI) rats and the CX3CR1 expression in spinal cord dorsal horn and dorsal root ganglia (DRG). METHOD: Seventy-two male SD rats were randomly divided into 4 groups: 1) Sham operation group (Sham); 2) Chronic constrictive injury group (CCI); 3) Curcumin treated group (Cur), administrated with curcumin 100 mg x kg(-1) x d(-1) ip for 14 days after CCI; 4) Solvent contrast group (SC), administrated with an equal volume of solvent for 14 days after CCI. Paw thermal withdrawal (PTWL) and paw mechanical withdrawal threshold (PMWT) were measured on 2 pre-operative and 1, 3, 5, 7, 10, 14 post-operative days respectively. The lumbar segments L4-5 of the spinal cord and the L4, L5 DRG were removed at 3, 7, 14 days after surgery. The expression of CX3CR1 was determined by immunohistochemical staining. RESULT: Compared with Sham group, PTWL and PMWT in CCI group were significantly lower on each post-operative day (P

A butachlor-degrading strain, designated FLY-8, was isolated from rice field soil and was identified as Paracoccus sp. Strain FLY-8 could degrade and utilize six chloroacetamide herbicides as carbon sources for growth, and the degradation rates followed the order alachloracetochlorpropisochlorbutachlorpretilachlormetolachlor. The influence of molecular structure of the chloroacetamide herbicides on the microbial degradation rate was first analyzed; the results indicated that the substitutions of alkoxymethyl side chain with alkoxyethyl side chain greatly reduced the degradation efficiencies; the length of amide nitrogen's alkoxymethyl significantly affected the biodegradability of these herbicides: the longer the alkyl was, the slower the degradation efficiencies occurred. The phenyl alkyl substituents have no obvious influence on the degradation efficiency. The pathway of butachlor complete mineralization was elucidated on the basis of the results of metabolite identification and enzyme assays. Butachlor was degraded to alachlor by partial C-dealkylation and then converted to 2-chloro-N-(2,6-dimethylphenyl)acetamide by N-dealkylation, which subsequently transformed to 2,6-diethylaniline, which was further degraded via the metabolites aniline and catechol, and catechol was oxidized through an ortho-cleavage pathway. This study highlights an important potential use of strain FLY-8 for the in situ bioremediation of chloroacetamide herbicides and their metabolite-contaminated environment.

To investigate the diversify of the nuclear pathway of c-Jun NH2-terminal kinases (JNK) during transient brain ischemia/reperfusion injury in hippocampal neuron apoptosis in spontaneously hypertensive rats (SHR) and to test whether the neuroprotection of curcumine on transient brain ischemia/reperfusion injury in SHR is related to the nuclear pathway of JNK.Male Wistar-Kyoto (WKY) rats and SHR were randomly divided into five groups (n = 6): WKY sham group (W-Sham), WKY ischemia/reperfusion group (W-I/ R), SHR sham group (S-Sham), SHR ischemia/reperfusion group (S-I/R) and SHR curcumine (a chinese traditional medicine)100 mg/kg treatment group (S-Cur), which were sacrificed at 2 h, 6 h, 24 h, 3 d and 7 d after reperfusion. Global brain ischemic model was established by 4-VO method. The TdT-mediated dUTP nick end labeling (TUNEL) method was used to detect the neuron apoptosis in hippocampal CA1 region. The immunohistochemical method was applied to investigate the expressions of c-jun and c-fos in hippocampal CA1 region.The expressions of apoptosis and c-jun and c-fos in CA1 region in S-Sham group, W-I/R group and S-I/R group were more than those in W-Sham group (P0.05), were significantly increased in S-I/R group than those in W-I/R group (P0.05), and were significantly decreased in S-Cur group than those in S-I/R group (P0.05).Neuronal apoptosis and the expressions of c-jun and c-fos are more in SHR hippocampal. Global brain ischemia/reperfusion injury induces more expressions of apoptosis in hippocampal neuron in SHR, and the more expressions of c-jun and c-fos may participate in that process. The neuroprotection of curcumine in SHR is related to c-jun and c-fos.

A novel non-sporulating, non-motile, catalase- and oxidase-positive, strictly aerobic, Gram-negative, rod-shaped bacterial strain, designated DCA-1T, was isolated from activated sludge collected from a butachlor wastewater treatment facility. The strain was able to degrade about 85 % of 100 mg butachlor l−1 within 5 days of incubation. Growth occurred in the presence of 0–6 % (w/v) NaCl [optimum, 1 % (w/v) NaCl] and at pH 5.5–9.0 (optimum, pH 7.0) and 15–35 °C (optimum, 25–30 °C). Vesicular internal membrane structures and photoheterotrophic growth were not observed. The major respiratory quinone was ubiquinone 10 (Q-10) and the major cellular fatty acids were C18 : 1ω7c and 11-methyl C18 : 1ω7c. The genomic DNA G+C content of strain DCA-1T was 62.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain DCA-1T was a member of the family Rhodobacteraceae and was related most closely to the type strain of Catellibacterium aquatile (96.5 % sequence similarity). The combination of phylogenetic analysis, phenotypic characteristics and chemotaxonomic data supports the suggestion that strain DCA-1T represents a novel species of the genus Catellibacterium, for which the name Catellibacterium caeni sp. nov. is proposed. The type strain is DCA-1T ( = CGMCC 1.7745T = DSM 21823T). In addition, based on the characterization data obtained in this study, it is proposed that Rhodobacter changlensis should be reclassified as Catellibacterium changlense comb. nov. (type strain JA139T = DSM 18774T = CCUG 53722T = JCM 14338T). An emended description of the genus Catellibacterium is also presented.

A synthetic pyrethroid (SP)-degrading bacterial strain, designated JZ-1T, was isolated from activated sludge of a SP-manufacturing wastewater treatment facility and studied using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JZ-1T belonged to the genus Sphingobium, showing highest sequence similarities to Sphingobium faniae DSM 21829T (98.6 %), Sphingobium cloacae JCM 10874T (98.5 %), Sphingobium vermicomposti DSM 21299T (97.4 %) and Sphingobium ummariense CCM 7431T (96.9 %). The polar lipid pattern, the presence of spermidine and ubiquinone Q-10, the predominance of the cellular fatty acids C18 : 1ω7c, C19 : 0 cyclo ω8c, 11 methyl C18 : 1ω7c, C16 : 0 and C14 : 0 2-OH, and the G+C content of the genomic DNA also supported the affiliation of the strain with the genus Sphingobium. Strain JZ-1T showed low DNA–DNA relatedness values with S. faniae DSM 21829T (30.2 %), S. cloacae JCM 10874T (23.3 %), S. vermicomposti DSM 21299T (10.9 %) and S. ummariense CCM 7431T (7.9 %). Based on its phylogenetic position and its phenotypic and genotypic properties, strain JZ-1T represents a novel species of the genus Sphingobium, for which the name Sphingobium wenxiniae sp. nov. is proposed. The type strain is JZ-1T ( = CGMCC 1.7748T = DSM 21828T).

The taxonomic status was determined of an aerobic, facultatively methylotrophic strain, JZL-4T, isolated from activated sludge. The cells were Gram-negative, asporogenous, colourless, motile, short rods. The strain utilized methanol, methylamine, formate and a variety of polycarbon compounds, but not methane, dichloromethane or CO2/H2, as carbon and energy sources. C1 compounds were assimilated via the isocitrate lyase-negative serine pathway. Optimal growth occurred at 30 °C, pH 6.5–7.5 and 0.5 % (w/v) NaCl. The major cellular fatty acids were C18 : 1ω7c and C18 : 0. The major phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and phosphatidylmonomethylethanolamine (PME); PME, the main phospholipid of strain JZL-4T, was absent or present in only minor amounts in Methylopila capsulata IM1T, Methylopila helvetica DM9T and Albibacter methylovorans DM10T. The major ubiquinone was Q-10. The DNA G+C content of strain JZL-4T was 70.4 mol% (T m). Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain showed high sequence similarities to M. capsulata IM1T (97.2 %), A. methylovorans DM10T (94.9 %) and M. helvetica DM9T (94.1 %), and showed less than 94 % similarity to strains of other species with validly published names. Strain JZL-4T had a low level of DNA–DNA relatedness (34 %) with M. capsulata IM1T. On the basis of phenotypic, genetic and phylogenetic data, strain JZL-4T is proposed to represent a novel species of the genus Methylopila, with the name Methylopila jiangsuensis sp. nov. The type strain is strain JZL-4T ( = ACCC 05406T = DSM 22718T = VKM B-2555T).

A Gram-staining-negative, oxidase-positive, catalase-positive, non-motile, non-spore-forming, and rod-shaped bacterium, designated BJQ-6T, was isolated from activated sludge of a waste-water treatment plant in Jiangsu Province, China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BJQ-6T belonged to the genus Rhodanobacter, and shared 98.7% similarity with Rhodanobacter lindaniclasticus LMG 18385T and

A metsulfuron-methyl-resistant yeast strain, JHL(T), was isolated from metsulfuron-methyl-contaminated soil collected in Jiangsu Province, China. Through morphological and physiological analysis as well as a molecular phylogenetic analysis based on the 26S rRNA gene D1/D2 region and internal transcribed spacer (ITS), this strain, which forms a clade with Candida vartiovaarae and a teleomorphic species, Williopsis saturnus, was revealed to represent a novel species in the genus Candida. The name Candida mengyuniae sp. nov. (type strain JHL(T)=CGMCC 2.3681(T)=CBS 10845(T)) is proposed for this novel species.