Impacts of PFOS, PFOA and their alternatives on the gut, intestinal barriers and gut-organ axis.

With the restricted use of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), a number of alternatives to PFOS and PFOA have attracted great interest. Most of the alternatives are still characterized by persistence, bioaccumulation, and a variety of toxicity. Due to the production and use of these substances, they can be detected in the atmosphere, soil and water body. They affect human health through several exposure pathways and especially enter the gut by drinking water and eating food, which results in gut toxicity. In this review, we summarized the effects of PFOS, PFOA and 9 alternatives on pathological changes in the gut, the disruption of physical, chemical, biological and immune barriers of the intestine, and the gut-organ axis. This review provides a valuable understanding of the gut toxicity of PFOS, PFOA and their alternatives as well as the human health risks of emerging contaminants.

Birnessite-Type MnO2 Modified Sustainable Biomass Fiber toward Adsorption Removal Heavy Metal Ion from Actual River Aquatic Environment.

In this work, a novel birnessite-type MnO2 modified corn husk sustainable biomass fiber (MnO2@CHF) adsorbent was fabricated for efficient cadmium (Cd) removal from aquatic environments. MnO2@CHF was designed from KMnO4 hydrothermally treated with corn husk fibers. Various characterization revealed that MnO2@CHF possessed the hierarchical structure nanosheets, large specific surface area, and multiple oxygen-containing functional groups. Batch adsorption experimental results indicated that the highest Cd (II) removal rate could be obtained at the optimal conditions of adsorbent amount of 0.200 g/L, adsorption time of 600 min, pH 6.00, and temperature of 40.0 °C. Adsorption isotherm and kinetics results showed that Cd (II) adsorption behavior on MnO2@CHF was a monolayer adsorption process and dominated by chemisorption and intraparticle diffusion. The optimum adsorption capacity (Langmuir model) of Cd (II) on MnO2@CHF was 23.0 mg/g, which was higher than those of other reported common biomass adsorbent materials. Further investigation indicated that the adsorption of Cd (II) on MnO2@CHF involved mainly ion exchange, surface complexation, redox reaction, and electrostatic attraction. Moreover, the maximum Cd (II) removal rate on MnO2@CHF from natural river samples (Xicheng Canal) could reach 59.2% during the first cycle test. This study showed that MnO2@CHF was an ideal candidate in Cd (II) practical application treatment, providing references for resource utilization of agricultural wastes for heavy metal removal.

Complex resistivity spectrum of pollutant soils with low-concentration heavy metals.

Soil contamination by heavy metals occurs globally, with varying degrees of severity, especially in agricultural fields. Investigating the frequency response characteristics of different types of heavy metal pollutants through induced polarization can provide valuable evidence for surveys based on this method. Soil specimens with varying low concentrations of copper (Cu), chromium (Cr), cadmium (Cd), and lead (Pb) heavy metals were prepared for this study, and parameters including complex resistivity, amplitude-frequency, and resistivity phase were measured. Our findings reveal the following trends: Complex resistivity decreases as heavy metal concentrations increase, demonstrating significant shifts within lower concentration ranges but presenting limitations for assessing pollution in high-concentration areas. Conversely, amplitude-frequency increases with higher heavy metal concentrations, displaying excellent performance in high-concentration scenarios. The differences in complex resistivity and amplitude-frequency among different types of heavy metal pollutants are distinct. In contrast, the absolute phase decreases with rising heavy metal concentrations. The resistivity phase spectra for various heavy metal pollutants exhibit unique patterns. For example, copper-contaminated soil exhibits phase peaks in the frequency range of 8-32 Hz, whereas chromium-contaminated soil shows phase peaks at 16-64 Hz. Cadmium-contaminated soil displays phase peaks ranging from 0.25 Hz to 2 Hz, while lead-contaminated soil exhibits phase peaks within the 0.5 Hz-4 Hz range. Leveraging the frequency range corresponding to phase peaks as an identification method for heavy metal pollution types proves effective. The frequency response characteristics of induced polarization vary significantly among different types and concentrations of heavy metal pollutants, providing important foundations for the application of induced polarization method in the field of heavy metal pollution detection.

A Study on the Efficacy of Ω Toenail Correction Treating Paronychia.

To study the clinical effects of Ω toenail correction in the treatment of paronychia. One hundred thirty-six cases of 130 patients during the period from August 2018 to August 2021 were treated with Ω toenail correction according to clinical stages, the clinical therapeutic effects of which were evaluated in terms of the operation time, the time to resume movement, treatment cycle, 1-y recurrence rate, and visual analogue scale (VAS) scores before and after treatment. The clinical efficacy was analyzed and compared of Ω toenail correction in treating paronychia of different clinical stages. It has been demonstrated that there was no significant difference in operation time, time to resume movement, treatment cycle and recurrence rate among different stages of paronychia, while there existed the significant difference (p < .05) in VAS score of resting-state pain before and after correction which stood at 6.43 ± 0.29 points with the after-treatment VAS scores at 1.10 ± 0.22. There is a statistical difference (p < .05) in VAS score of movement-evoked pain between before and after treatment. The VAS scores of movement-evoked pain stood at 7.55 ± 0.42, which is in contrast with the after-treatment VAS at 1.74 ± 0.93. It has been concluded that Ω toenail correction characterized by easy operation can relieve the pain immediately, which can achieve satisfactory clinical efficacy for treating paronychia of different stages.

Serum hsa_circ_0000615 is a prognostic biomarker of sorafenib resistance in hepatocellular carcinoma.

BACKGROUND: Circular RNAs (circRNAs) can shape tumor progression and chemoresistance. How specific circRNAs shape hepatocellular carcinoma (HCC) chemoresistance, however, remains to be fully elucidated. METHODS: In total, serum samples were collected from 202 HCC patients that had completed four sorafenib chemotherapy cycles. Serum hsa_circ_0000615 levels in these patients were quantified via quantitative real-time polymerase chain reaction (qRT-PCR), with demographic details and survival outcomes being recorded for subsequent analyses. RESULTS: We found hsa_circ_0000615 to be significantly upregulated in chemoresistant HCC patients relative to chemosensitive patients, with such upregulation being positively correlated with disease stage. Moreover, the area under the curve (AUC) value for hsa_circ_0000615 was moderately good, and high levels of hsa_circ_0000615 expression were associated with shorter overall survival among chemoresistant HCC patients. CONCLUSION: Our results highlight hsa_circ_0000615 as a promising driver of sorafenib resistance in HCC patients, highlighting it as a promising target for the treatment of this deadly cancer type.

NOD2 is involved in regulating odontogenic differentiation of DPSCs suppressed by MDP through NF-κB/p65 signaling.

Dental pulp stem cells (DPSCs) are well known for their capable of both self-renewal and multilineage differentiation. Dental tissue diseases, include caries, are often accompanied by inflammatory microenvironment, and muramyl dipeptide (MDP) is involved in the inflammatory stimuli to influence the differentiation of DPSCs. Nucleotide-binding oligomerization domain 2 (NOD2), a member of the cytosolic Nod-like receptor (NLR) family, plays a key role in inflammatory homeostasis regulation, but the role of NOD2 in DPSCs differentiation under inflammatory is still unclear. In this study, we identified that MDP suppressed odontogenic differentiation of DPSCs via NOD2/ NF-κB/p65 signaling pathway. Alizarin red staining and ALP activity showed the odontogenic differentiation was suppressed by MDP in a concentration-dependent manner, and the expression of dentin differentiation marker protein dentin matrix protein 1 (DMP-1) and dentin Sialophosphoprotein (DSPP) also indicated the same results. The expression of NOD2 increased gradually with the concentration of MDP as well as the phosphorylation and nuclear translocation of p65, which meant NF-κB signaling pathway was activated. Further, the interference of NOD2 inhibited the phosphorylation and nuclear translocation of p65 and reversed the MDP-mediated decrease of odontoblast differentiation of DPSCs. Our study showed that MDP can inhibit the odontoblast differentiation of DPSCs in a concentration-dependent manner. The NF-κB signaling pathway was activated by increasing expression of NOD2. Interference of NOD2 reversed the negative ability odontoblast differentiation of DPSCs in the inflammatory environment. Our study might provide a theoretical basis for the clinical treatment for dentinogenesis of DPSCs.

Ecological risk assessment for organophosphate esters in the surface water from the Bohai Sea of China using multimodal species sensitivity distributions.

Organophosphate esters (OPEs) as the foremost substitutes of brominated flame retardants have been ubiquitously found in the aquatic environment around the world. However, the information on the community-level risks induced by OPEs to the marine ecosystem remains scarce. This study adopted ten commonly used species sensitivity distribution (SSD) parametric statistical approaches coupled with the acute-to-chronic transformation for the toxicity data to fit the sensitivity distributions of different species to four major OPE congeners including triethyl phosphate (TEP), tri-n-butyl phosphate (TnBP), tri(2-chloroethyl) phosphate (TCEP), and tris(1-chloro-2-propyl) phosphate (TCPP) in the surface water of the Bohai Sea. All SSD models except Exponential for TnBP, TCEP, and TCPP fitted well the chronic toxicity data for the four OPE congeners. Discrepancies appeared among the best fitting models for different congeners, which also happened to the fitting results from the multiple SSD models for each congener. Based on the best fitting models, the hazard concentrations corresponding to the cumulative probability of 5% were 3.58 mg/L, 0.116 mg/L, 1.30 mg/L, and 1.44 mg/L for TEP, TnBP, TCEP, and TCPP, respectively. The risks induced by the four OPE congeners to the Bohai Sea ecosystem were negligible during the monitoring period because of both the risk quotients and the hazard indexes far <0.1. This study drew a clear picture of the joint ecological risks of TEP, TnBP, TCEP, and TCPP to the Bohai Sea environment. The application of multimodal SSD statistical methods will benefit the accurate derivation of water quality criteria and the community-level ecological risk assessment for pollutants.

Pterostilbene exert an anti-arthritic effect by attenuating inflammation, oxidative stress, and alteration of gut microbiota.

Pterostilbene is a revesterol analog with a long bioavailability and having potent anti-inflammatory activity in animal studies. In this study, we tried to scrutinize the anti-arthritic effect of pterostilbene against complete Freund's adjuvant (CFA)-induced arthritis model in rats. CFA was used for induction of the RA, and rats were divided into groups depending on different doses of pterostilbene given. Hepatic, antioxidant, rheumatoid factor (RF), myeloperoxidase (MPO), inflammatory cytokines, anti-collagen(C)II-Ig, inflammatory mediators, and gut microbiota were estimated. Pterostilbene significantly (p < .001) decreased the paw swelling, arthritic score, and increased the body weight. Besides altered the antioxidant, inflammatory mediators, anti-collagen (C)II Ig, and inflammatory cytokines. Furthermore, pterostilbene treatment helps to restore the ecosystem of gut microbiota in rats by reducing the relative abundance of Helicobacter, Desulfovibrio, Lachnospiraceae, and Mucispirillium. Based on the findings, we can say that pterostilbene has an anti-arthritic effect via suppressing inflammatory responses and altering intestinal bacteria. PRACTICAL APPLICATIONS: Arthritis is the painful disease and affected most of the people worldwide. In this experimental study, we estimated the anti-arthritic effect of pterostilbene against CFA-induced arthritis in rats. Pterostilbene noticeably suppressed the paw thickness, arthritic score, and organ index. Pterostilbene substantially altered the oxidative stress and inflammatory reaction. Pterostilbene considerably modulated the gut microbiota, suggesting the anti-arthritic effect.

Bone Mesenchymal Stem Cells Promote Extracellular Matrix Remodeling of Degenerated Nucleus Pulposus Cells via the miR-101-3p/EIF4G2 Axis.

The etiology of lumbocrural pain is tightly concerned with intervertebral disk degeneration (IDD). Bone mesenchymal stem cell (BMSC)-based therapy bears potentials for IDD treatment. The properties of microRNA (miRNA)-modified BMSCs may be altered. This study investigated the role and mechanism of BMSCs promoting extracellular matrix (ECM) remodeling of degenerated nucleus pulposus cells (NPCs) via the miR-101-3p/EIF4G2 axis. NPCs were collected from patients with IDD and lumbar vertebral fracture (LVF). The expressions of miR-101-3p and ECM-related proteins, Collagen-I (Col-I) and Collagen-II (Col-II), were detected using the reverse transcription-quantitative polymerase chain reaction. The expressions of Col-I and Col-II, major non-collagenous component Aggrecan, and major catabolic factor Matrix metalloproteinase-13 (MMP-13) were detected using Western blotting. BMSCs were cocultured with degenerated NPCs from patients with IDD. Viability and apoptosis of NPCs were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry. After the degenerated NPCs were transfected with the miR-101-3p inhibitor, the expressions of ECM-related proteins, cell viability, and apoptosis were detected. The targeting relationship between miR-101-3p and EIF4G2 was verified. Functional rescue experiments verified the effects of miR-101-3p and EIF4G2 on ECM remodeling of NPCs. Compared with the NPCs of patients with LVF, the degenerated NPCs of patients with IDD showed downregulated miR-101-3p, Col-II, and Aggrecan expressions and upregulated MMP-13 and Col-I expressions. BMSCs increased the expressions of miR-101-3p, Aggrecan, and Col-II, and decreased the expressions of MMP-13 and Col-I in degenerated NPCs. BMSCs enhanced NPC viability and repressed apoptosis. Downregulation of miR-101-3p suppressed the promoting effect of BMSCs on ECM remodeling. miR-101-3p targeted EIF4G2. Downregulation of EIF4G2 reversed the inhibiting effect of the miR-101-3p inhibitor on ECM remodeling. In conclusion, BMSCs increased the miR-101-3p expression in degenerated NPCs to target EIF4G2, thus promoting the ECM remodeling of NPCs.

Proton Conduction of Nafion Hybrid Membranes Promoted by NH3-Modified Zn-MOF with Host-Guest Collaborative Hydrogen Bonds for H2/O2 Fuel Cell Applications.

It is of great significance to develop creative proton exchange membrane materials for proton exchange membrane fuel cells (PEMFCs). The strategy of doping metal-organic frameworks (MOFs) with guest molecules into the Nafion matrix is adopted to improve the electrochemical performance of Nafion hybrid membranes. Various and abundant hydrogen bonds can make a tremendous contribution to the proton conduction of hybrid membranes. In this work, we used high proton-conducting Zn-MOFs with the characteristics of host-guest collaborative hydrogen bonds as the filler to prepare Zn-MOF/Nafion hybrid membranes. Alternating current (AC) impedance tests show that when the doping amount of Zn-MOF is 5%, the proton conductivity reaches 7.29 × 10-3 S·cm-1, being 1.87 times that of the pure Nafion membrane at 58% relative humidity (RH) and 80 °C. In an attempt to prove the promotion effect of guest NH3 on proton conductivity of Nafion hybrid membranes, Zn-MOF-NH3 was filled into the Nafion matrix. Under the same conditions, its proton conductivity reaches the maximum value of 2.13 × 10-2 S·cm-1, which is 5.47 times that of the pure Nafion membrane. Zn-MOF-NH3/Nafion-5 was used to fabricate a proton exchange membrane for application in H2/O2 fuel cells. The maximum power density of 212 mW cm-2 and a current density of 630 mA cm-2 reveal a respectable single cell performance. This study provides a promising method for optimizing the structure of MOF proton conductors and inspires the preparation of high-performance Nafion hybrid membranes.

HBV-Pol is crucial for HBV-mediated inhibition of inflammasome activation and IL-1ß production.

BACKGROUND & AIMS: Hepatitis B virus (HBV) infection is the most critical factor underlying liver cirrhosis and hepatocellular carcinoma worldwide. IL-1ß and IL-18, generated by activation of the inflammasome/caspase-1 signaling pathway, play important roles in the control and clearance of HBV. However, the specific relationship between the inflammasome response and IFN-α resistance or viral persistence is yet to be established. METHODS: Blood samples of patients and supernatant fractions of HBV cell lines were collected for analysis and the effects on inflammasome activation and IL-1ß production evaluated via enzyme-linked immunosorbent assay (ELISA), western blot, quantitative RT-PCR and immunofluorescence. RESULTS: IL-1ß and IL-18 levels produced in sera of IFN-α non-responders were significantly lower than those of responders and normal donors. Additionally, expression of IL-1ß and inflammasome components was decreased in peripheral blood mononuclear cells (PBMC) of non-responders, compared with those of responders. In vitro experiments on HepG2, HepG2.2.15 and HepAD38 cell lines showed that HBV induces a significant decrease in IL-1ß production through inhibiting activation of the NF-κB signaling and inflammasome/caspase-1 pathways. And hepatitis B virus polymerase (HBV-Pol) appeared crucial for these inhibitory effects of HBV. CONCLUSION: IL-1ß production is suppressed in HBV carriers and IFN-α non-responders. HBV induces a significant decrease in IL-1ß production through inhibiting the NF-κB signaling and inflammasome pathways, for which HBV-Pol is a crucial requirement. Trial approval number: 20 173 402.

Hepatitis B core antigen can regulate NLRP3 inflammasome pathway in HepG2 cells.

Hepatitis B virus (HBV) has four open reading frames (ORFs) of which ORF C is consists of the pre Core and Core genes encodes the Hepatitis B core antigen (HBcAg) and Hepatitis B e antigen (HBeAg). Studies have shown that HBeAg significantly inhibits the NLRP3 inflammasome activation and interleukin-1ß (IL-1ß) production. However, the role of HBcAg and ORF C proteins (in this paper, ORF C proteins = HBcAg + HBeAg) were remain unclear. Our study aims to assess whether HBcAg and ORF C proteins can affect the NLRP3 inflammasome pathway. Vectors expressing ORF C proteins and HBcAg were designed and transfected into HepG2 cells. And then, cells were stimulated with lipopolysaccharide (LPS). Activation of the NLRP3 inflammasome and the levels of IL-1ß and IL-18 were evaluated by Western blot analysis, quantitative reverse-transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and immunofluorescence. The expression of NLRP3 and IL-1ß peaked when HepG2 cells were stimulated with 1000 ng/mL LPS for 18 to 24 hours. HBcAg, but not ORF C proteins, promoted LPS-induced NLRP3 inflammasome activation and IL-1ß production. These findings provide a novel mechanism on how the HBV causes liver inflammation and may provide insights into the search for new therapeutic strategies.

HEV ORF3 downregulates TLR7 to inhibit the generation of type I interferon via impairment of multiple signaling pathways.

Hepatitis E is the most common type of acute hepatitis prevalent worldwide. The open reading frame 3 protein of HEV (HEV ORF3) is proposed to create a favorable environment for viral replication and pathogenesis. However, the mechanisms by which HEV overcomes the effects of host immunity, particularly the role of ORF3, remain to be established. Expression of IFNα and IFNß in supernatant and cell samples was examined via ELISA and quantitative RT-PCR. The protein levels of specific signaling factors in cells overexpressing HEV ORF3 were examined via western blot. Analyses of cells transfected with vectors expressing ORF3 demonstrated that HEV ORF3 significantly impairs the generation of endogenous type I interferon through downregulating TLR3 and TLR7 as well as their corresponding downstream signaling pathways. Moreover, inhibition of NFκB, JAK/STAT and JNK/MAPK signaling pathways contributed significantly to suppression of increased levels of TLR7. Levels of p-P65, p-STAT1 and p-JNK were markedly impaired in ORF3-expressing cells, even upon treatment with the respective agonists. HEV ORF3 inhibits the production of endogenous type I interferon through downregulation of TLR3 and TLR7. Furthermore, suppression of TLR7 is achieved through impairment of multiple signaling pathways, including NFκB, JAK/STAT and JNK/MAPK.

Exploration of ZEA cytotoxicity to mouse endometrial stromal cells and RNA-seq analysis.

Because zearalenone (ZEA) causes harmful influence to animals and widely exists in the world, the researches on ZEA have never stopped. However, the mechanisms of ZEA on proliferation, cycle, and apoptosis in endometrial stromal cells (ESCs) remain poorly defined. Therefore, the purpose of our study was to explore the effects of ZEA to ESCs and demonstrate them by transcriptomic analysis. The results showed that after ZEA treatment, ESCs appeared numerous adverse reactions, and the phenomena of cell viability decrease, DNA replication block and apoptosis were detected by flow cytometry, Annexin V-FITC/PI double-staining method, TUNEL assay, and so on. Then, RNA-seq approach was adopted to prove the validity of above experiments, as expected, the results from different expression genes, gene ontology terms, and KEGG pathway were all consistent with those. Overall, the results suggested that ZEA could cause a series of reactions by cytotoxicity to mouse ESCs, meanwhile there must be some substances and mechanisms protect cells against cytotoxicity damage.

Genetic Variants in the STMN1 Transcriptional Regulatory Region Affect Promoter Activity and Fear Behavior in English Springer Spaniels.

Stathmin 1 (STMN1) is a neuronal growth-associated protein that is involved in microtubule dynamics and plays an important role in synaptic outgrowth and plasticity. Given that STMN1 affects fear behavior, we hypothesized that genetic variations in the STMN1 transcriptional regulatory region affect gene transcription activity and control fear behavior. In this study, two single nucleotide polymorphisms (SNPs), g. -327 A>G and g. -125 C>T, were identified in 317 English Springer Spaniels. A bioinformatics analysis revealed that both were loci located in the canine STMN1 putative promoter region and affected transcription factor binding. A statistical analysis revealed that the TT genotype at g.-125 C>T produced a significantly greater fear level than that of the CC genotype (P < 0.05). Furthermore, the H4H4 (GTGT) haplotype combination was significantly associated with canine fear behavior (P < 0.01). Using serially truncated constructs of the STMN1 promoters and the luciferase reporter, we found that a 395 bp (-312 nt to +83 nt) fragment constituted the core promoter region. The luciferase assay also revealed that the H4 (GT) haplotype promoter had higher activity than that of other haplotypes. Overall, our results suggest that the two SNPs in the canine STMN1 promoter region could affect canine fear behavior by altering STMN1 transcriptional activity.

Exploration of Bcl-2 family and caspases-dependent apoptotic signaling pathway in Zearalenone-treated mouse endometrial stromal cells.

Zearalenone (ZEA) is a nonsteroidal estrogenic mycotoxin found in several food commodities worldwide. Although the toxicity of ZEA have been widely studied in a number of cell types, the mechanistic role of ZEA on apoptosis of endometrial stromal cells (ESCs) remains poorly understood. The objective of this study was to determine the effects of ZEA on apoptosis of mouse ESCs and explore the signaling pathway underlying the cytotoxicity of ZEA. The results showed that ZEA treatment caused obvious apoptosis in ESCs as determined by the flow cytometry and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay. Immunoblotting and real-time quantitative polymerase chain reaction (RT-qPCR) revealed that ZEA treatment increased the ratio of Bax/Bcl-2. The enzymatic activity assays revealed that caspases-3 and caspase-9 were activated by ZEA treatment in a dose-dependent manner. In addition, flow cytometry show that the apoptotic percentages of cells pretreated with Z-VAD-FMK and Z-LEHD-FMK were markedly reduced compared to the ZEA-treated cells. Overall, the results suggested that ZEA induced obvious apoptosis in ESCs via a Bcl-2 family and caspases-dependent signaling pathway.

Sirtuin Inhibition Adversely Affects Porcine Oocyte Meiosis.

Sirtuins have been implicated in diverse biological processes, including oxidative stress, energy metabolism, cell migration, and aging. Here, we employed Sirtuin inhibitors, nicotinamide (NAM) and Sirtinol, to investigate their effects on porcine oocyte maturation respectively. The rate of polar body extrusion in porcine oocytes decreased after treatment with NAM and Sirtinol, accompanied with the failure of cumulus cell expansion. We further found that NAM and Sirtinol significantly disrupted oocyte polarity, and inhibited the formation of actin cap and cortical granule-free domain (CGFD). Moreover, the abnormal spindles and misaligned chromosomes were readily detected during porcine oocyte maturation after treatment with NAM and Sirtinol. Together, these results suggest that Sirtuins are involved in cortical polarity and spindle organization in porcine oocytes.

Treatment of thoracolumbar vertebrate fracture by transpedicular morselized bone grafting in vertebrae for spinal fusion and pedicle screw fixation.

To enhance the fusion of graft bone in thoracolumbar vertebrae and minimize the postoperative loss of correction, short-segment pedicle screw fixation was reinforced with posterior morselized bone grafting in vertebrae for spinal fusion in patients with thoracolumbar vertebrate fractures. Seventy patients with thoracolumbar vertebrate fractures were treated by short-segment pedicle screw fixation and were randomly divided into two groups. Fractures in group A (n=20) were reinforced with posterior morselized bone grafting in vertebrae for spinal fusion, while patients group B (n=50) did not receive the morselized bone grafting for bone fusion. The two groups were compared in terms of kyphotic deformity, anterior vertebral height, instrument failure and neurological functions after the treatment. Frankel grading system was used for the evaluation of neurological evaluation and Denis scoring scale was employed for pain assessment. The results showed that the kyphosis correction was achieved in both group A and group B (group A: 6.4 degree; group B: 5.4 degree)/ At the end of follow-up, kyphosis correction was maintained in group A but lost in group B (P=0.0001). Postoperatively, greater anterior height was achieved in group A than in group B (P<0.01). During follow-up study, anterior vertebral height was maintained only in Group A (P<0.001). Both group A and group B showed good Denis pain scores (P1 and P2) but group A outdid group B in terms of control of severe and constant pain (P4 and P5). By Frankel criteria, the changes in neurological functions in group A was better than those of group B (P<0.001). It is concluded that reinforcement of short-segment pedicle fixation with morselized bone grafting for the treatment of patients with thoracolumbar vertebrae fracture could achieve and maintain kyphosis correction, and it may also increase and maintain anterior vertebral height. Morselized bone grafting in vertebrae offers immediate spinal stability in patients with thoracolumbar vertebrate fractures, decreases the instrument failure and provides better postoperative pain control than without the morselized bone grafting.

Treatment of Thoracolumbar Vertebrate Fracture by Transpedicular Morselized Bone Grafting in Vertebrae for Spinal Fusion and Pedicle Screw Fixation / 华中科技大学学报（医学）（英德文版）

To enhance the fusion of graft bone in thoracolumbar vertebrae and minimize the postoperative loss of correction, short-segment pedicle screw fixation was reinforced with posterior moselizee bone grafting in vertebrae for spinal fusion in patients with thoracrolumbar vertebrate fractures. Seventy patients with thoracrolumbar vertebrate fractures were treated by short-segment pedicle screw fixation and were randomly divided into two groups. Fractures in group A (n=20) were reinforced with posterior morselized bone grafting in vertebrae for spinal fusion, while patients group B (n=50) did not receive the morselized bone grafting for bone fusion. The two groups were compared in terms of kyphotic deformity, anterior vertebral height, instrument failure and neurological functions after the treatment. Frankel grading system was used for the evaluation of neurological evaluation and Denis scoring scale was employed for pain assessment. The results showed that the kyphosis correction was achieved in both group A and group B (group A: 6.4 degree; group B: 5.4 degree)/At the end of follow-up, kyphosis correction was maintained in group A but lost in group B (P=0.0001). Postoperatively, greater anterior height was achieved in group A than in group B (P<0.01). During follow-up study, anterior vertebral height was maintained only in Group A (P<0.001). Both group A and group B showed good Denis pain scores (P1 and P2) but group A outdid group B in terms of control of severe and constant pain (P4 and P5). By Frankel criteria, the changes in neurological functions in group A was better than those of group B (P<0.001). It is concluded that reinforcement of short-segment pedicle fixation with morselized bone grafting for the treatment of patients with thoracolumbar vertebrae fracture could achieve and maintain kyphosis correction, and it may also increase and maintain anterior vertebral height. Morselized bone grafting in vertebrae offers immediate spinal stability in patients with thoracolumbar vertebrate fractures, decreases the instrument failure and provides better postoperative pain control than without the morselized bone grafting.

Construction of a new food-grade expression system for Bacillus subtilis based on theta replication plasmids and auxotrophic complementation.

A new food-grade expression system was constructed for Bacillus subtilis based on replicative food-grade expression plasmids and auxotrophic complementation. The food-grade B. subtilis host FG01 was created by knockout of the dal locus from the chromosome of B. subtilis 168. Two food-grade expression plasmids pXFGT03 and pXFGT05 were constructed by combining a novel theta-type Bacillus replicon with the B. subtilis endogenous gene dal and P43 promoter; while pXFGT05 was derived from pXFGT03 by deletion of two open reading frames (ORFs) from the original replicon. Upon transformation of FG01 with pXFGT03 or pXFGT05, the host phenotype was complemented on Luria-Bertani agar plates by the plasmid-coded dal gene, which served as a food-grade selection marker for recombinants. Results showed that deletion of the two ORFs had no impact on plasmid replication. A reporter gene bgaB was cloned into pXFGT03 and pXFGT05, respectively, under control of the P43 promoter, and it was successfully expressed in this food-grade expression system. Segregational stabilities of two recombinant plasmids were investigated, and they were fully stable.