First report of Pectobacterium carotovorum and Pectobacterium brasiliense causing bacterial soft rot of bok choy in Taiwan.

Bok choy (Brassica rapa var. chinensis) is one of the most popular leafy green vegetables in Asia (Wang et al. 2019; Zhang et al. 2014). In May 2022, disease resembling bacterial soft rot was observed in a commercial greenhouse located in Xiluo, Yunlin County, Taiwan. Affected plants exhibited maceration, primarily close to the base of the plants (Fig. S1). Almost all bok choy plants (about 1,800 plants in total) on site were symptomatic. Macerated tissues were collected from six plants. The samples were homogenized in 10 mM MgCl2 and bacteria were isolated on nutrient agar (NA) by streak plating. After 1 day of culturing at 28°C, creamy-white, round colonies were consistently grown on all the plates, and six strains (Br1 to Br6) were obtained; each isolated from a different plant. The strains were able to ferment glucose and induced maceration on potato tuber slices (Schaad et al. 2001) but could not produce indigoidine on NGM medium (NA added with glycerol and MnCl2; Lee and Yu 2006). The DNA samples of these strains were tested with Pectobacterium-specific primers Y1 and Y2 (Darrasse et al. 1994) and all samples produced the expected amplicon. To identify the isolated pathogens, 1,592-bp sequences concatenated from fragments of the leuS (452 bp), dnaX (492 bp), and recA (648 bp) genes (GenBank accession nos. OP360013-OP360021) were obtained for each strain as previously described (Portier et al. 2019). Three genotypes were detected, the sequences of strains Br1, Br2, Br4, and Br5 were identical, while strains Br3 and Br6 each belong to a different genotype. The sequence identity between Br3 and Br6 was 98.2%. The concatenated sequences (dnaX-leuS-recA), along with those of type strains from known Pectobacterium species, were subjected to maximum likelihood analysis. The reconstructed trees showed that strains Br1, Br2, Br4, and Br5 grouped with P. carotovorum CFBP2046T (Fig. S2); the sequence identity between the isolated strains and the type strain was 98.7%. Strains Br3 and Br6 clustered with P. brasiliense CFBP6617T (Fig S2); the sequence identity between CFBP6617T and Br3 and Br6 were 97.5% and 98.4%, respectively. The six strains were inoculated onto 55-day-old bok choy plants using previously described prick inoculation methods (Wei et al. 2019). Autoclaved toothpicks, each carrying 9.3 x 106- 5.6 x 107 cfu of bacteria, were used to inoculate the base of plant leaves. All six strains were tested, and each strain had three replicates. Plants in the control group were stabbed with bacteria-free toothpicks. The plants were enclosed in clear plastic bags during the assay to maintain humidity and kept in a growth chamber (27/25°C day/night; 14-h photoperiod). After 1 d, all inoculated plants produced soft rot symptoms resembling those observed in the sampling site. No noticeable differences were observed among symptoms produced by different strains. The controls were symptomless. One strain was re-isolated from each treatment group and their identity were confirmed by sequencing the dnaX gene. All re-isolated strains shared the same sequences with those of the original strains tested. This is the first report of P. brasiliense and P. carotovorum causing bacterial soft rot of bok choy in Taiwan. Importantly, the findings showed that different Pectobacterium species and genotypes could induce symptoms on a crop in the same facility at the same time, highlighting the potential complexity of interactions among different soft rot bacteria in the environment.

First report of bacterial soft rot on Epipremnum aureum caused by Pectobacterium aroidearum in Taiwan.

Pothos (Epipremnum aureum) is an Araceae foliage plant with great ornamental values, which has long been enjoyed by consumers (Chen et al. 2010). In September 2021, pothos showing soft rot symptoms were found in 2 nurseries in Taichung, Taiwan. The petioles of the infected plants were macerated; some lesions extended to the leaves (Figure S1). The disease incidence was 50% in one nursery and 37.5% in the other; two and three plants were respectively collected from the two sites. Macerated tissues were homogenized in 10 mM MgCl2 and the samples were observed microscopically without dyeing. Motile, rod-shaped bacteria were observed in the samples, and the bacteria were isolated onto nutrient agar (NA) and grown at 28°C for 2 days. Fast-growing, round, creamy colonies were isolated from all 5 plants. One strain was isolated from each plant and the strains were named Ea1 to Ea5. The bacteria could ferment glucose and induce maceration on potato tuber slices (Schaad et al. 2001), but did not produce indigoidine on NGM medium (Lee and Yu 2006) and were tested negative for phosphatase activity (Schaad et al. 2001). The bacteria's DNA samples were tested using primers specific to Pectobacterium (Y1/Y2; Darrasse et al. 1994). The expected 434-bp amplicon was amplified in all five strains. Multilocus sequence analysis was conducted as previously described (Portier et al. 2019). A concatenated sequence (1,592 bp) comprising partial dnaX (492 bp), leuS (452 bp) and recA (648 bp) sequences was obtained for each strain. Two genotypes were detected among the strains; Ea1 and Ea2 belonged to one genotype (i.e., they had identical sequences), while Ea3, Ea4 and Ea5 belonged to the other (GenBank accession nos. OK416015-OK416020). Phylogenetic analysis was conducted using these data and those of representative strains of known Pectobacterium species (Klair et al. 2022). A maximum-likelihood tree showed that Ea1 to Ea5 clustered with P. aroidearum CFBP8168T (Figure S2). Sequence comparison (Table S1) showed that the similarity between the two genotypes' concatenated sequences was 99.1% (Ea1 vs. Ea3; 1,578/1,592 bp); Ea1 and Ea3 shared 99.2% and 99.3% sequence similarity with P. aroidearum CFBP8168T, respectively. The sequences obtained in this work were searched against GenBank and all of their top hits were those of strains belonging to P. aroidearum (supplementary information). Koch's Postulates were fulfilled by stab inoculating cutting-propagated pothos (8-cm tall) using toothpicks carrying bacteria grown on NA. The pathogen loads used were estimated by suspending cells (attached to individual toothpicks) in 10 mM MgCl2 and spread-plating them onto NA (after dilution); the loads were 5.5 x 106 - 2.2 x 107 CFU. Three plants were inoculated for each strain (3 petioles per plant). Control plants were stabbed with sterile toothpicks. Each plant was then bagged and placed in a growth chamber (28°C; 14 h light). After 24 h, all inoculated plants produced symptoms resembling those found in the nurseries, and the controls did not. For every treatment group, a strain was re-isolated onto NA; each of them shared the same recA sequence with the original strain inoculated. This is first report of P. aroidearum causing pothos soft rot in Taiwan. Local nurseries often grow pothos and other Araceae plants together in humid areas. Since other Araceae species are also known to be susceptible to P. aroidearum (Xu et al. 2020), growers should be cautious of the pathogen's spread across hosts.

Long-term clinical benefit of Peg-IFNα and NAs sequential anti-viral therapy on HBV related HCC.

Analysis of the value of long-term antiviral therapy using sequential Peg-IFN therapy and nucleos(t)ide analogues (NAs) improves the prognosis of HBV-related HCC. HBV-related HCC patients were classified into sequential therapy with Peg-IFNα-2a and NAs, and NAs therapy alone. All patients were followed up for 5 years. The survival rate, HCC recurrence rate, Child-Pugh score, and side effects of drugs were evaluated. Firstly, the early and late cumulative survival rate was higher in patients receiving antiviral therapy compared with the control patients (p<0.05). Patients receiving sequential therapy with Peg-IFNα-2a and NAs showed a higher late cumulative survival rate and significantly reduced early and late recurrence rate, compared to those in the NA-alone group (p<0.05). Single NAs therapy only reduced the late recurrence rate in HCC-patients. Secondly, NAs therapy significantly increased the Child-Pugh score after five years of therapy (five-year therapy 7.03±1.50 vs. initial score 6.63±0.85; p<0.05), whereas the sequential therapy with Peg-IFNα-2a and NAs did not greatly alter the Child-Pugh score (6.88±1.26; p>0.05). Compared to the control patients, patients receiving antiviral therapy (NAs alone or sequential therapy with Peg-IFNα-2a and NAs) exhibited a significantly decreased Child-Pugh score (p<0.05). Compared to NAs alone, sequential therapy with Peg-IFNα-2a and NAs provided a more efficient strategy for improving both the five-year survival rate and the two-year or five-year recurrence rate in patients.

Ergosterol-loaded poly(lactide-co-glycolide) nanoparticles with enhanced in vitro antitumor activity and oral bioavailability.

AIM: Ergosterol is a plant sterol with anti-tumor and anti-angiogenic activities, but is poorly soluble. In this study, we attempted to enhance its anti-tumor action and oral bioavailability via poly(lactide-co-glycolide) (PLGA) nanoparticle encapsulation. METHODS: Ergosterol-loaded PLGA nanoparticles (NPs/Erg) were prepared using the emulsion/solvent evaporation technique. Their physicochemical properties were characterized, and their cytotoxicity against human cancer cell lines was evaluated with MTT assay. The pharmacokinetics and tissue distribution of NPs/Erg were investigated in rats and mice, respectively. RESULTS: NPs/Erg were spherical in shape with a particle size of 156.9±4.8 nm and a Zeta potential of -19.27±1.13 mV, and had acceptable encapsulation efficiency and loading capacity. NPs/Erg exerted much stronger cytotoxicity against human cancer cells than the free ergosterol, and showed significantly reduced IC50 values (14.69±0.48 µg/mL in glioma U251 cells; 9.43±0.52 µg/mL in breast cancer MCF-7 cells; 4.70±0.41 µg/mL in hepatoma HepG2 cells). After oral administration of a single dose in rats, NPs/Erg displayed a prolonged plasma circulation with a 4.9-fold increase of oral bioavailability compared with the free ergosterol. After mice received NPs/Erg, the ergosterol in NPs/Erg was rapidly distributed in stomach, kidneys, liver, brain, spleen, and virtually non-existent in heart and lungs. The presence of NPs/Erg in brain was particularly improved compared with the free ergosterol. CONCLUSION: The PLGA nanoparticles serve as a promising carrier for the poorly soluble ergosterol and significantly improve its bioavailability, biodistribution and in vitro anti-tumor activities.

Preparation of amino-modified carbon quantum dots-ZnO/cellulose nanofiber multifunctional hydrogel: Enhanced adsorption synergistic photoreduction and reversible fluorescence response visual recognition of Cr(VI).

This work innovatively used cellulose nanofibers as a photocatalyst carrier, which could recycle nano-photocatalysts and minimize nanoparticle aggregation. The morphology, structures, chemical composition, optical-electronic properties and photocatalytic performance of amino-modified carbon quantum dots-ZnO/cellulose nanofiber (N-CQDs-ZnO/CNF: ZCH-2) hydrogel were characterized by SEM, TEM, BET, EDS, XRD, FTIR, UV-vis, XPS, PL and other techniques. The mechanism of Cr(VI) adsorption synergistic photoreduction by ZCH-2 was discussed in detail. The results showed that the prepared ZCH-2 had excellent removal performance for Cr(VI). After 120 min of adsorption and 40 min of photoreduction, the removal efficiency of Cr(VI) was 98.9 %. Compared with ZnO/CNF hydrogel, the adsorption performance of ZCH-2 increased by 268 % and the photoreduction performance increased by 116 %. The adsorption of Cr(VI) by ZCH-2 was controlled by electrostatic attraction and chemical adsorption. The photoreduction kinetic constant of ZCH-2 was 0.106 min-1, which was 8.9 times that of ZnO/CNF hydrogel. The N-CQDs in ZCH-2 could form N-CQDs-metal complexes with Cr(VI), resulting in fluorescence quenching, so Cr(VI) could be visually identified by fluorescence changes. This study provides a new idea for the design and optimization of a new multifunctional hydrogel with efficient adsorption-photoreduction-fluorescence recognition.

Effects of naturally aged microplastics on the distribution and bioavailability of arsenic in soil aggregates and its accumulation in lettuce.

Naturally aged microplastics (NAMPs) and arsenic (As) have been reported to coexist in and threaten potentially to soil-plant ecosystem. The research explored the combined toxic effects of NAMPs and As to lettuce (Lactuca sativa L.) growth, and the distribution, accumulation and bioavailability of As in soil aggregates. The As contaminated soil with low, medium and high concentrations (L-As, M-As, H-As) were treated with or without NAMPs, and a total of six treatments. The results displayed that, in comparison to separate treatments of L-As and M-As, the presence of NAMPs increased the total biomass of lettuce grown at these two As concentrations by 68.9 % and 55.4 %, respectively. Simultaneous exposure of NAMPs and L-As or M-As led to a decrease in As content in shoot (0.45-2.17 mg kg-1) and root (5.68-14.66 mg kg-1) of lettuce, indicating an antagonistic effect between them. In contrast, co-exposure to H-As and NAMPs showed synergistic toxicity, and the leaf chlorophyll and nutritional quality of lettuce were also reduced. NAMPs altered the ratio of different soil aggregate fractions and the distribution of bioavailable As within them, which influenced the absorption of As by lettuce. In conclusion, these direct observations assist us in enhancing the comprehend of the As migration and enrichment characteristics in soil-plant system under the influence of NAMPs.

MiR-146a-5p Promotes Dental Stem Cells Osteo/odontogenic Differentiation through NF-Kappa B Signaling Pathway by Targeting TRAF6.

OBJECTIVE: To screen miRNAs that could simultaneously regulate osteo/odontogenic differentiation of multiple stem cells, including dental pulp stem cells (DPSCs), stem cells from the apical papilla (SCAPs) and periodontal ligament stem cells (PDLSCs). METHODS: Differentially expressed miRNAs analysis on three miRNA microarrays data of dental stem cells undergoing osteo/odontogenic differentiation (GSE138180, GSE154466 and GSE159508) was performed, and miR-146a-5p were identified by bioinformatic prediction, dual-luciferase reporter assay and quantitative real-time polymerase chain reaction (PCR). In addition, differentially expressed genes between miR-146a-5p overexpressed group and control group (GSE79341) were applied for KEGG pathways enrichment analysis. RESULTS: MiR-146a-5p expression increased in the osteo/odontogenic differentiation of DPSCs, SCAPs and PDLSCs. Tumour necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) was identified as the target gene of miR-146a-5p. Furthermore, miR-146a-5p could influence the NF-Kappa B signalling pathway. CONCLUSION: This study suggests that miR-146a-5p could promote differentiation in multiple dental stem cells through the NF-Kappa B signalling pathway by targeting TRAF6.

[Changes of the upper airway in children with Class â ¡ mandibular retrusion and snoring during night before and after functional treatment by sagittal-guidance Twin-block appliance].

PURPOSE: To study the changes of dimension and morphology of upper airway in children with ClassⅡ mandibular retrusion after functional treatment by sagittal-guidance Twin-block appliance. METHODS: Cone-beam CT (CBCT) data of upper airway of the subjects were measured by Dolphin 11.5 software and Mimics 17.0 software , and the changes of dimension and morphology of upper airway before and after functional treatment with sagittal-guidance Twin-block(SGTB) appliance were compared. SPSS 16.0 software package was used for data processing. RESULTS: After functional treatment，the volume of total upper airway，nasopharynx airway, oropharynx airway, the sectional area of tip of the epiglotti(TE), the lateral diameter of TE, the base of the epiglottis(EB) significantly increased (P<0.05) in children with SGTB appliance. CONCLUSIONS: SGTB functional treatment is effective in the treatment of skeletal ClassⅡ mandibular retrusion of children by increase of the upper airway and improvement of respiration．.

Intraperitoneal administration of biocompatible hyaluronic acid hydrogel containing multi-chemotherapeutic agents for treatment of colorectal peritoneal carcinomatosis.

Colorectal peritoneal carcinomatosis (CRPC) is an advanced stage of colorectal cancer (CRC), which significantly decreases patient survival and quality of life. Here, the naturally occurring polysaccharide hyaluronic acid (HA) was used to prepare an injectable hydrogel and simultaneously deliver 5-fluorouracil (5-FU), cisplatin (DDP) and paclitaxel (PTX) microspheres for intraperitoneal CRPC chemotherapy. The drug-loaded HA hydrogel released the drugs in a sustained manner, and showed low toxicity both in vitro and in a mouse model of CRPC. Furthermore, direct injection of the drug-loaded HA hydrogel in the abdominal cavity of tumor-bearing mice significantly decreased tumor growth and liver/lung metastasis, along with decreasing the volume of ascites and inhibiting local intestinal infiltration of the tumor cells. Therefore, this novel multi-drug hydrogel delivery system may effectively clear CRPC tumors without any adverse effects when used in intraperitoneal chemotherapy.

Therapeutic efficacy of thermosensitive Pluronic hydrogel for codelivery of resveratrol microspheres and cisplatin in the treatment of liver cancer ascites.

Ascites constitutes the most frequent decompensating event in patients with advanced liver cancer and is associated with poor quality of life and high mortality. Intraperitoneal chemotherapy appears to be a reliable treatment strategy for advanced liver cancer ascites. However, the rapid metabolism of drugs and ascites dilution limits the efficacy of chemotherapeutics. Therefore, the present study aimed to develop a novel thermosensitive hydrogel drug system for targeted therapy of advanced hepatocellular carcinoma (HCC) ascites through intraperitoneal administration. The system was prepared by blending resveratrol (RES) microspheres and cisplatin (DDP) into thermosensitive Pluronic F127 hydrogel. The in vitro anti-tumor activity against H22 cells indicated that the prepared drug system could initiate apoptosis and induce cell cycle arrest at the G1 phase. The mice model of ascites with advanced HCC was established to validate the therapeutic potential of the F127 hydrogel drug system in vivo. The results revealed that intraperitoneal administration of F127 hydrogel drug could significantly inhibit the number of ascites, the proliferation of tumor cells, micro-angiogenesis, and prolong the survival of mice, thus, augmenting the efficacy of intraperitoneal chemotherapy. Moreover, immunohistochemical staining revealed that the F127 hydrogel drug system was safe and presented low toxicity to major vital organs. Collectively, this study highlights the clinical application potential of the F127 hydrogel drug delivery system.

Self-assembled dihydroartemisinin nanoparticles as a platform for cervical cancer chemotherapy.

Dihydroartemisinin (DHA) is a potent anti-cancer drug that has limited clinical applications due to poor water solubility and low bioavailability. We designed a biodegradable poly(ethylene glycol) methyl ether-poly(ε-caprolactone) (MPEG-PCL) micelle carrier for DHA using the self-assembly method. The DHA/MPEG-PCL nanoparticles were spherical with an average particle size of 30.28 ± 0.27 nm, and released the drug in a sustained manner in aqueous solution. The drug-loaded nanoparticles showed dose-dependent toxicity in HeLa cells by inducing cycle arrest and apoptosis. Furthermore, compared to free DHA, the DHA/MPEG-PCL nanoparticles showed higher therapeutic efficacy and lower toxicity in vivo, and significantly inhibited tumor growth and prolonged the survival of tumor-bearing nude mice. In addition, the tumor tissues of the DHA/MPEG-PCL-treated mice showed a marked decline in the in situ expression of proliferation and angiogenesis markers. Taken together, the self-assembled DHA/MPEG-PCL nanoparticles are a highly promising delivery system for targeted cancer treatment.

[Gnathology in implant-supported fixed restoration in edentulous mandible].

OBJECTIVE: This study aims to investigate the occlusal and myoelectric characteristics of implant-supported fixed denture in the mandibular region and provide reference for the design of fixed restoration. METHODS: Sixty edentulous patients with implant-supported fixed denture were selected and divided into three groups: group A, 20 cases with implant-supported fixed restoration in the maxillary region; group B, 20 cases with natural dentition, and group C, 20 cases with removable partial denture. The T-scan 8.0 digital occlusion analysis system was used to evaluate the occlusal characteristics of patients in the three groups at intercuspal, protrusion, and left and right lateral positions. Electromyography was used to analyze the myoelectric amplitude and bilateral asymmetry index of the anterior temporalis and masseter of the three groups in different states such as resting and clenching. The relationship between occlusion and myoelectricity was also investigated. RESULTS: In the occlusion analysis by T-scan, the occlusion time, the balance of left and right bite force, the left and right asymmetry of the occlusion center, the trajectory of central occlusion force, and the disclusion time were higher in group C than in groups A and B (P<0.05). No significant differences were observed in the anterior and posterior asymmetry of the occlusion center and percentage of bite force at anterior region among the three groups. In the analysis of myoelectricity, the myoelectric amplitude at resting state and the asymmetry index of masticatory muscles in group C were higher than those in groups A and B (P<0.05). The myoelectric amplitude during clenching in groups A and B groups was higher than that in group C (P<0.05). CONCLUSIONS: In implant-supported fixed restoration at edentulous mandibular, when maxillary includes the removable partial denture, degree of occlusal instability and left and right asymmetry of occlusion center are greater than those with the natural dentition and implant-supported fixed denture at maxillary. The myoelectricity is closely related to occlusion. The removable partial denture can increase the myoelectric activity and reduce the potential of the masticatory muscle. The asymmetry of bilateral myoelectricity is related to the occlusion imbalance.

Application of mesoporous carbon-polymer monolith for the extraction of phenolic acid in food samples.

In this study, activated carbon-organic polymer (AC-polymer) extraction columns were prepared via microwave-assisted polymerization. The AC-polymer was applied in polymer monolith microextraction (PMME) of phenolic acids (PAs) in food samples. Structural characterization using Raman and Fourier transform infrared (FTIR) spectroscopy revealed the successful incorporation of AC into polymer monolith. In contrast to neat polymer in which low extraction efficiency (17.0-57.7%) was observed, the AC-polymer (8 mg AC, 4 cm column length) afforded better extraction efficiency for PAs ranging from 75.8 to 99.8% for intra-day with less than 1.4% relative standard deviations (RSDs) and inter-day precision ranging from 74.3 to 100.1% (<2.7% RSDs). The extraction efficiency for column-to-column were found to be in the range of 72.8-100.5% (<2.0% RSDs) (n = 3). Under the optimized conditions, the AC-polymer monolithic column was successfully applied in PMME of PAs in fruit wine and cranberry juice samples at concentrations of <4.8 mg L-1 and 26.54 mg L-1, respectively.

Regulatory polymorphism of CXCL10 rs1439490 in seronegative occult hepatitis C virus infection.

AIM: To examine the relationship between the single nucleotide polymorphism CXCL10 rs1439490 and seronegative occult hepatitis C virus (HCV) infection (OCI). METHODS: One hundred and three cases of seronegative OCI and 155 cases of seropositive chronic HCV infection (CHC) were diagnosed at five Liver Centers in Northeastern China, from 2012 to 2016. CXCL10 rs1439490, rs1440802, and IL-28B rs12979860 were analyzed by sequencing. Serum CXCL10 was measured by ELISA. Intrahepatic CXCL10 was determined by quantitative PCR and immunohistochemical semi-quantitative scoring. Liver necroinflammation and fibrosis were scored according to the METAVIR system. RESULTS: CXCL10 rs1439490 G/G was more prevalent in OCI patients (n = 93/103; 90.3%) than in CHC patients (n = 116/155; 74.8%; P = 0.008). OCI patients had lower serum CXCL10 levels than CHC patients (192.91 ± 46.50 pg/mL vs 354.78 ± 102.91 pg/mL, P < 0.0001). Of IL-28B rs12979860 C/C patients, OCI patients with rs1439490 G/G had lower serum and liver levels of CXCL10 and lower levels of liver necroinflammation and fibrosis than non-G/G patients. OCI patients had higher alanine aminotransferase normalization rates after Peg-interferon treatment than CHC patients (P < 0.05) and serum CXCL10 decreased significantly (P < 0.0001). Liver necroinflammation and fibrosis were alleviated in 8 OCI patients after treatment. Multivariate analysis indicated that rs1439490 G/G significantly influenced the occurrence of OCI in HCV infection (OR = 0.31, 95%CI: 0.15-0.66, P = 0.002). CONCLUSION: CXCL10 rs1439490 G/G is positively associated with OCI in HCV infection and antiviral outcome.

Aqueous biphasic systems containing PEG-based deep eutectic solvents for high-performance partitioning of RNA.

In this work, 16 kinds of novel deep eutectic solvents (DESs) composed of polyethylene glycol (PEG) and quaternary ammonium salts, were coupled with Aqueous Biphasic Systems (ABSs) to extract RNA. The phase forming ability of ABSs were comprehensively evaluated, involving the effects of various proportions of DESs' components, carbon chain length and anions species of quaternary ammonium salts, average molecular weights of PEG and inorganic salts nature. Then the systems were applied in RNA extraction, and the results revealed that the extraction efficiency values were distinctly enhanced by relatively lower PEG content in DESs, smaller PEG molecular weights, longer carbon chain of quaternary ammonium salts and more hydrophobic inorganic salts. Then the systems composed of [TBAB][PEG600] and Na2SO4 were utilized in the influence factor experiments, proving that the electrostatic interaction was the dominant force for RNA extraction. Therefore, back-extraction efficiency values ranging between 85.19% and 90.78% were obtained by adjusting the ionic strength. Besides, the selective separation of RNA and tryptophane (Trp) was successfully accomplished. It was found that 86.19% RNA was distributed in the bottom phase, while 72.02% Trp was enriched in the top phase in the novel ABSs. Finally, dynamic light scattering (DLS) and transmission electron microscope (TEM) were used to further investigate the extraction mechanism. The proposed method reveals the outstanding feasibility of the newly developed ABSs formed by PEG-based DESs and inorganic salts for the green extraction of RNA.

Detection of Imjin Virus and Seoul Virus in Crocidurine Shrews in Shandong Province, China.

INTRODUCTION: Recently, hantaviruses have been discovered in insectivores in Europe, Asia, Africa, and North America. Imjin virus (MJNV) was first isolated from the lung tissues of Ussuri white-toothed shrew (Crocidura lasiura) from South Korea in 2009. We aim to detect the species and prevalence of insectivore- and rodent-borne hantaviruses in shrews and rodents. MATERIALS AND METHODS: Shrews and rodents were captured in Jiaonan County of Shandong Province, China, in 2014. RT-PCR was used to amplify viral RNA of Hantavirus species, including insectivore-borne Imjin virus (MJNV), rodent-borne Hantaan virus (HTNV), and Seoul virus (SEOV) from shrews and rodents. RESULTS AND DISCUSSION: We found that MJNV infected 10.7% (19/178) of Crocidura shrews, but it infected none of rodents (0/475); we also found that 2 of 178 (1.1%) Crocidura shrews were PCR positive to SEOV. This study indicated that the major animal hosts of Imjin virus are shrews, and rodent-borne SEOV can infect shrews.

A novel polymeric ionic liquid-coated magnetic multiwalled carbon nanotubes for the solid-phase extraction of Cu, Zn-superoxide dismutase.

A novel magnetic adsorbent, benzyl groups functionalized imidazolium-based polymeric ionic liquid (PIL)-coated magnetic multiwalled carbon nanotubes (MWCNTs) (m-MWCNTs@PIL), has been successfully synthesized and applied for the extraction of Cu, Zn-superoxide dismutase (Cu, Zn-SOD). The m-MWCNTs@PIL were characterized by X-ray diffraction (XRD), Fourier transform infrared spectrometry (FT-IR), thermal gravimetric analysis (TGA), field emission scanning electron microscopy (FESEM), vibrating sample magnetometer (VSM) and zeta-potential nanoparticles. In this method, the m-MWCNTs@PIL could interact with Cu, Zn-SOD through hydrogen bonding, π-π and electrostatic interactions. The extraction performance of the m-MWCNTs@PIL in the magnetic solid-phase extraction (MSPE) procedure was investigated, coupled with the determination by UV-vis spectrophotometer. Compared with m-MWCNTs@IL and m-MWCNTs, the m-MWCNTs@PIL exhibited the highest extraction capacity of 29.1 mg/g for Cu, Zn-SOD. The adsorbed Cu, Zn-SOD remained high specific activity after being eluted from m-MWCNTs@PIL by 1 moL/L NaCl solution. Besides, the m-MWCNTs@PIL could be easily recycled and successfully employed in the extraction of Cu, Zn-SOD from real samples. Under the optimal conditions, the precision, repeatability and stability of the proposed method were investigated and the RSDs were 0.29%, 1.68% and 0.54%, respectively. Recoveries were in the range of 82.7-102.3%, with the RSD between 3.47% and 5.35%. On the basis of these results, the developed method has great potential in the extraction of Cu, Zn-SOD or other analytes from biological samples.

[Morphological characteristics of upper alveolar bone around central incisor in Class II division 1 and division 2 malocclusion on cone-beam CT].

PURPOSE: During orthodontic treatment, the incisors in patients with Class II division 1 and division 2 malocclusion showed different clinical features, which determine the different ways to move the teeth to the suitable position. This study analyzed the morphology of alveolar bone around upper central incisor with cone-beam CT (CBCT). METHODS: The sample consisted of 20 normal malocclusion and 40 patients with Class II malocclusion （division 1 20 cases and division 2 20 cases）. CBCT images before orthodontic treatment were reconstructed by INVIVO 5.0 image processing software. Paired t test was used for statistical analysis with SPSS 19.0 software package. RESULTS: At the upper central incisors, the alveolar thickness between normal malocclusion and Class II division 1 and division 2 malocclusion had significant difference except the palate thickness between normal malocclusion and Class II division 2 malocclusion (P<0.05). CONCLUSIONS: Orthodontist should consider the relationship between the teeth position and alveolar bone shape comprehensively, choose the correct plan in order to avoid root desorption, alveolar bone loss, fenestration and other adverse reactions.

[Preparation of 8-chloro-adenosine long circulation liposomes and its pharmacokinetics in rats].

AIM: To prepare 8-chloro-adenosine (8-Cl-A) long circulation liposomes with high entrapped efficiency and prolonged action-time of 8-Cl-A in vivo. METHODS: To prepare 8-Cl-A long circulation liposomes of nanometer size by improved multiple emulsion. The entrapped efficiency, size and size distribution of 8-Cl-A long circulation liposomes were determined, and its pharmacokinetics in rats was evaluated. RESULTS: The entrapped efficiency of 8-Cl-A long circulation liposomes was 62.70% and mean diameter of the liposomes was 79.9 nm. The pharmacokinetics studies indicated that 8-Cl-A long circulation liposomes showed higher drug concentration and larger AUC values than that of 8-Cl-A after iv to rats. CONCLUSION: 8-Cl-A long circulation liposomes could prolong the action-time of 8-Cl-A in vivo.

[A study on the thickness of buccal bone in the mandible of different vertical facial type in adults using cone-beam CT].

PURPOSE: To provide some references of using micro-implant anchorage in clinical orthodontic treatment, the thickness of buccal bone of mandible of different vertical facial type in adults with cone-beam CT (CBCT) were measured. METHODS: Initial 3-dimensional images of 45 adult patients (15 men, 30 women)were reoriented by using a standardized protocol, and divided into 3 groups by angle of mandibular plane (G1: high angle, 15 patients; G2: average angle, 15 patients; G3: low angle, 15 patients). After signing the informed consent form, three measurement points were defined at 4, 6, and 8mm from the alveolar crest in each measurement area. Statistical analysis was performed with SPSS11.0 software package. RESULTS: The thickness of buccal bone in mandible was thinner in G1 than in G2, and was thinnest in G3. The cortical bone thickness was thickest at the 8 mm level and thinnest at the 4mm level. CONCLUSIONS: The study suggested that the cortical bone thickness at 4mm level from the alveolar crest was safe for implantation. It is more stable when the distance of implants was more away from the alveolar crest. Patients with low mandibular plane angle should use self-tapping micro-implant anchorage. Supported by Nantong Science and Technology Fund (HS2013033).