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Chao Zhang, Dike Ruan, Qing He, Tianyong Wen, Pushan YangDepartment of Orthopedic Surgery, Navy General Hospital, Beijing, People's Republic of ChinaAbstract: Diffuse idiopathic skeletal hyperostosis (DISH) is considered an underdiagnosed and mostly asymptomatic nonprimary osteoarthritis. The etiology of DISH remains unknown and the validated diagnostic criteria are absent. This condition is still recognized radiologically only. Rarely, large projecting anterior osteophytes result in esophageal impingement and distortion leading to dysphagia. We report the case of progressive dysphagia and neck pain due to DISH of the cervical spine in a 70-year-old man, which was surgically removed with excellent postoperative results and complete resolution of symptoms. Imaging studies, surgical findings, and histopathological examinations were used to support the diagnosis. The patient was successfully treated with total excision of the anterior osteophytes with no evidence of recurrence 12 months after surgery. In this report, we also discuss the clinical features and perioperative considerations in combination with a literature review. Our patient illustrates that clinicians should be aware of this rare clinical manifestation as the presenting feature of DISH in cervical spine. Surgical decompression through osteophytectomy is effective for patients who fail conservative treatment.Keywords: diffuse idiopathic skeletal hyperostosis, cervical spine, dysphagia

Melioidosis, an infection caused by the facultative intracellular pathogen Burkholderia pseudomallei, has been classified as an emerging disease with the number of patients steadily increasing at an alarming rate. B. pseudomalleipossess various virulence determinants that allow them to invade the host and evade the host immune response, such as the type III secretion systems (TTSS). The products of this specialized secretion system are particularly important for the B. pseudomallei infection. Lacking in one or more components of the TTSS demonstrated different degrees of defects in the intracellular lifecycle of B. pseudomallei. Further understanding the functional roles of proteins involved in B. pseudomallei TTSS will enable us to dissect the enigma of B. pseudomallei-host cell interaction. In this study, BipC (a translocator), which was previously reported to be involved in the pathogenesis of B. pseudomallei, was further characterized using the bioinformatics and molecular approaches. The bipCgene, coding for a putative invasive protein, was first PCR amplified from B. pseudomallei K96243 genomic DNA and cloned into an expression vector for overexpression in Escherichia coli. The soluble protein was subsequently purified and assayed for actin polymerization and depolymerization. BipC was verified to subvert the host actin dynamics as demonstrated by the capability to polymerize actin in vitro. Homology modeling was also attempted to predict the structure of BipC. Overall, our findings identified that the protein encoded by the bipC gene plays a role as an effector involved in the actin binding activity to facilitate internalization of B. pseudomalleiinto the host cells.

Pinus is an important economic tree species, but pine wilt disease (PWD) seriously threatens the survival of pine trees. PWD caused by Bursaphelenchus xylophilus is a major quarantine disease worldwide that causes significant economic losses. However, more information about its molecular pathogenesis is needed, resulting in a lack of effective prevention and treatment measures. In recent years, effectors have become a hot topic in exploring the molecular pathogenic mechanism of pathogens. Here, we identified a specific effector, BxNMP1, from B. xylophilus . In situ hybridization experiments revealed that BxNMP1 was specifically expressed in dorsal gland cells and intestinal cells, and RT-qPCR experiments revealed that BxNMP1 was upregulated in the early stage of infection. The sequence of BxNMP1 was different in the avirulent strain, and when BxNMP1 -silenced B. xylophilus was inoculated into P. thunbergii seedlings, the disease severity significantly decreased. We demonstrated that BxNMP1 interacted with the thaumatin-like protein PtTLP-L2 in P. thunbergii . Additionally, we found that the β-1,3-glucanase PtGLU interacted with PtTLP-L2. Therefore, we hypothesized that BxNMP1 might indirectly interact with PtGLU through PtTLP-L2 as an intermediate mediator. Both targets can respond to infection, and PtTLP-L2 can enhance the resistance of pine trees. Moreover, we detected increased salicylic acid contents in P. thunbergii seedlings inoculated with B. xylophilus when BxNMP1 was silenced or when the PtTLP-L2 recombinant protein was added. In summary, we identified a key virulence effector of PWNs, BxNMP1. It positively regulates the pathogenicity of B. xylophilus and interacts directly with PtTLP-L2 and indirectly with PtGLU. It also inhibits the expression of two targets and the host salicylic acid pathway. This study provides theoretical guidance and a practical basis for controlling PWD and breeding for disease resistance.

Bursaphelenchus xylophilus is the pathogen of pine wilt disease, which can devastate the pine forest ecosystem. Usually, plant cells generate reactive oxygen species (ROS) as a defensive substance or signalling molecules to resist the infection of nematodes. However, little is known about how B. xylophilus effectors mediate the plant ROS metabolism. Here, we identified a pioneer B. xylophilus Prx3-interacting effector 1 (BxPIE1) expressed in the dorsal gland cells and the intestine. Silencing of the BxPIE1 gene resulted in reduced nematode reproduction and a delay in disease progression during parasitic stages, with the upregulation of pathogenesis-related (PR) genes PtPR-3 (class Ⅳ chitinase) and PtPR-9 (peroxidase). The protein-protein interaction assays further demonstrated that BxPIE1 interacts with a Pinus thunbergii class III peroxidase (PtPrx3), which produces H 2 O 2 under biotic stress. The expression of BxPIE1 and PtPrx3 was upregulated during the infection stage. Furthermore, BxPIE1 effectively inhibited H 2 O 2 generating from class III peroxidase and ascorbate can recover the virulence of siBxPIE1-treated B. xylophilus by scavenging H 2 O 2 . Taken together, BxPIE1 is an important virulence factor, revealing a novel mechanism utilized by nematodes to suppress plant immunity., (© 2024 John Wiley & Sons Ltd.)

Four new phomalones A-D (1-4), together with five known analogues (5-9) were isolated from the deep-sea-derived fungus Trichobotrys effuse FS522. Their structures of the new compounds established by analysis of their NMR and HR-ESI-MS spectroscopic data, and the absolute configurations of 2 was determined by electronic circular dichroism (ECD) calculations. compounds 4, 6 and 8 substantially inhibited the production of nitric oxide (NO) with IC 50 values of 4.64, 13.90, and 34.07 μM., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

We measured winter and summer soil organic carbon (SOC) contents in two typical coastal wetlands, the Spartina alterniflora salt marsh and the non-vegetation mudflat, on the south side of the Chuandong River Estuary in Yancheng, Jiangsu Province. We investigated the spatiotemporal variations of soil organic carbon contents and its driving factors. The results showed that SOC content ranged from 0.75 to 2.38 g·kg -1 in the mudflat area and from 2.07 to 18.59 g·kg -1 in the S. alterniflora salt marsh area, showing a decreasing trend towards the sea. The SOC content in the S. alterniflora salt marsh area was approximately 2.5 to 3.5 times of that in the mudflat area. Within a depth range of 1 m, there was no vertical variation in SOC content in the mudflat area, but an increasing and then decreasing pattern in the S. alterniflora marsh area with the peak occurring in the depth range of 20 to 30 cm. Soil organic carbon content exhibited significant seasonal difference, with higher value in summer than in winter. The summer SOC content was 5% to 10% higher than that in winter in the S. alterniflora marsh area, while it was 43% higher in summer than in winter in the mudflat area. In the S. alterniflora marsh area, soil organic carbon content was positively correlated with soil moisture and salinity, but negatively correlated with sediment particle size. In contrast, there was no significant correlation between soil organic carbon content and soil physicochemi-cal factors in the mudflat area. Those results indicated that the correlation between various soil physicochemical factors and SOC is established on the basis of vegetation cover in coastal wetlands. Our findings could provide valuable insights for the conservation of blue carbon ecosystems in coastal wetlands in China.

Lipase is involved in lipid hydrolysis, which is related to nematodes' energy reserves and stress resistance. However, the role of lipases in Bursaphelenchus xylophilus, a notorious plant-parasitic nematode responsible for severe damage to pine forest ecosystems, remains largely obscure. Here, we characterized a class III lipase as a candidate effector and named it BxLip-3. It was transcriptionally up-regulated in the parasitic stages of B. xylophilus and specifically expressed in the oesophageal gland cells and the intestine. In addition, BxLip-3 suppressed cell death triggered by the pathogen-associated molecular patterns PsXEG1 and BxCDP1 in Nicotiana benthamiana, and its Lipase-3 domain is essential for immunosuppression. Silencing of the BxLip-3 gene resulted in a delay in disease onset and increased the activity of antioxidant enzymes and the expression of pathogenesis-related (PR) genes. Plant chitinases are thought to be PR proteins involved in the defence system against pathogen attack. Using yeast two-hybrid and co-immunoprecipitation assays, we identified two class I chitinases in Pinus thunbergii, PtChia1-3 and PtChia1-4, as targets of BxLip-3. The expression of these two chitinases was up-regulated during B. xylophilus inoculation and inhibited by BxLip-3. Overall, this study illustrated that BxLip-3 is a crucial virulence factor that plays a critical role in the interaction between B. xylophilus and host pine., (© 2023 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Pine wilt disease (caused by the nematode Bursaphelenchus xylophilus) is extremely harmful to pine forests in East Asia. As a low-resistance pine species, Pinus thunbergii is more vulnerable to pine wood nematode (PWN) than Pinus densiflora and Pinus massoniana. Field inoculation experiments were conducted on PWN-resistant and -susceptible P. thunbergii, and the difference in transcription profiles 24 h after inoculation was analyzed. We identified 2603 differentially expressed genes (DEGs) in PWN-susceptible P. thunbergii, while 2559 DEGs were identified in PWN-resistant P. thunbergii. Before inoculation, DEGs between PWN-resistant and PWN-susceptible P. thunbergii were enriched in the REDOX (Oxidation-Reduction) activity pathway (152 DEGs), followed by the oxidoreductase activity pathway (106 DEGs). After inoculation with PWN, however, the opposite was observed; DEGs were enriched in the oxidoreductase activity pathway (119 DEGs), followed by the REDOX activity pathway (84 DEGs). Before inoculation, according to the metabolic pathway analysis results, we found more genes upregulated in phenylpropanoid metabolic pathways and enriched in lignin synthesis pathways; cinnamoyl-CoA reductase-coding genes related to lignin synthesis were upregulated in PWN-resistant P. thunbergii and downregulated in PWN-susceptible P. thunbergii, and the lignin content was always higher in resistant than in susceptible P. thunbergii. These results reveal distinctive strategies of resistant and susceptible P. thunbergii in dealing with PWN infections., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: To explore the technical aspects of the accuracy of cervical pedicle screw placement with O-arm guidance., Methods: The clinical data of 21 patients who underwent cervical pedicle screw fixation by O-arm real-time guidance from December 2015 to January 2020 were analyzed retrospectively. There were 15 males and 6 females, aged from 29 to 76 years old with an average of (45.3±11.5) years. The postoperative CT scan was utilized to evaluate the placement of the pedicle screw and classified according to the Gertzbein and Robbins classification., Results: A total of 132 pedicle screws were implanted in 21 patients, 116 at C 3 -C 6 and 16 at C 1 and C 2 . According to Gertzbein & Robbins classification, the overall breach rates were found to be 11.36% (15/132) with 73.33% (11 screws) Grade B, 26.67% (4 screws) Grade C, and no Grade D or E screw breaches. There were no pedicle screw placement related complications at final follow-up., Conclusion: The application of O-arm real-time guidance technology can make cervical pedicle screw placement reliable. High accuracy and better intra-operative control can increase surgeon's confidence in using cervical pedicle instrumentation. Considering the high-risk nature of anatomical area around cervical pedicle and the possibility of catastrophic complications, the spine surgeon should have sufficient surgical skills, experience, ensures stringent verification of the system, and never relies solely on the navigation system.

Pine wilt disease, caused by Bursaphelenchus xylophilus , results in tremendous economic loss in conifer production every year. To disturb the host immune responses, plant pathogens secrete a mass of effector proteins that facilitate the infection process. Although several effectors of B. xylophilus have been identified, detailed mechanisms of their functions remain largely unexplored. Here, we reveal two novel B. xylophilus Kunitz effectors, named BxKU1 and BxKU2, using different infection strategies to suppress immunity in Pinus thunbergii . We found that both BxKU1 and BxKU2 could suppress PsXEG1-triggered cell death and were present in the nucleus and cytoplasm in Nicotiana benthamiana . However, they had different three-dimensional structures and various expression patterns in B. xylophilus infection. In situ hybridization experiments showed that BxKU2 was expressed in the esophageal glands and ovaries, whereas BxKU1 was only expressed in the esophageal glands of females. We further confirmed that the morbidity was significantly decreased in P. thunbergii infected with B. xylophilus when BxKU1 and BxKU2 were silenced. The silenced BxKU2I, but not BxKU1 , affected the reproduction and feeding rate of B. xylophilus . Moreover, BxKU1 and BxKU2 targeted to different proteins in P. thunbergii , but they all interacted with thaumatin-like protein 4 (TLP4) according to yeast two-hybrid screening. Collectively, our study showed that B. xylophilus could incorporate two Kunitz effectors in a multilayer strategy to counter immune response in P. thunbergii , which could help us better understand the interaction between plant and B. xylophilus .

The pinewood nematode, Bursaphelenchus xylophilus , has been determined as one of the world's top ten plant-parasitic nematodes. It causes pine wilt, a progressive disease that affects the economy and ecologically sustainable development in East Asia. B. xylophilus secretes pathogenic proteins into host plant tissues to promote infection. However, little is known about the interaction between B. xylophilus and pines. Previous studies reported transthyretin proteins in some species and their strong correlation with immune evasion, which has also been poorly studied in B. xylophilus . In this study, we cloned and functionally validated the B. xylophilus pathogenic protein BxTTR-52, containing a transthyretin domain. An in situ hybridization assay demonstrated that BxTTR-52 was expressed mainly in the esophageal glands of B. xylophilus . Confocal microscopy revealed that BxTTR-52-RFP localized to the nucleus, cytoplasm, and plasma membrane. BxTTR-52 recombinant proteins produced by Escherichia coli could be suppressed by hydrogen peroxide and antioxidant enzymes in pines. Moreover, silencing BxTTR-52 significantly attenuated the morbidity of Pinus thunbergii infected with B. xylophilus . It also suppressed the expression of pathogenesis-related genes in P. thunbergii . These results suggest that BxTTR-52 suppresses the plant immune response in the host pines and might contribute to the pathogenicity of B. xylophilus in the early infection stages.

Iron deficiency causes chlorosis and growth inhibition in Cinnamomum camphora , an important landscaping tree species. Siderophores produced by plant growth-promoting rhizobacteria have been widely reported to play an indispensable role in plant iron nutrition. However, little to date has been determined about how microbial siderophores promote plant iron absorption. In this study, multidisciplinary approaches, including physiological, biochemical and transcriptome methods, were used to investigate the role of deferoxamine (DFO) in regulating Fe availability in C. camphora seedlings. Our results showed that DFO supplementation significantly increased the Fe 2+ content, SPAD value and ferric-chelate reductase (FCR) activity in plants, suggesting its beneficial effect under Fe deficiency. This DFO-driven amelioration of Fe deficiency was further supported by the improvement of photosynthesis. Intriguingly, DFO treatment activated the metabolic pathway of glutathione (GSH) synthesis, and exogenous spraying reduced glutathione and also alleviated chlorosis in C. camphora . In addition, the expression of some Fe acquisition and transport-related genes, including CcbHLH , CcFRO6 , CcIRT2, CcNramp5 , CcOPT3 and CcVIT4 , was significantly upregulated by DFO treatment. Collectively, our data demonstrated an effective, economical and feasible organic iron-complexing agent for iron-deficient camphor trees and provided new insights into the mechanism by which siderophores promote iron absorption in plants.

Managing indoor ozone levels is important because ozone is a hazardous pollutant that has adverse effects on human health. Coffee is a popular daily beverage, and thus, coffee beans and spent coffee grounds are common in many places such as offices, homes, aircraft, cafeterias, and such. The most common material used to remove ozone is activated carbon which can be made from coffee beans or spent coffee grounds with proper activation processes. This paper presents a novel idea: to remove ozone at the level of an indoor environment using unactivated coffee products. This paper examines the ozone removal efficiency and the ozone deposition velocity at 130 ppb ozone for two types of coffee: solid coffee (powder) and liquid coffee (beverage). The activated carbon, the deionized water, and the seawater are also included for comparison and validation purposes. The tests show that the fine coffee powder has a removal efficiency of 58.5% and a deposition velocity of 0.62 cm/s. The liquid coffee has a removal efficiency of 34.4% and a deposition velocity of 0.23 cm/s. The chemical inspections indicate that the oxidation reactions with the carbohydrates in solid coffee and the metal/mineral elements in liquid coffee are responsible for ozone removal. These results have confirmed that ozone removal via coffee is effective, controlling indoor air quality by coffee products is thus becoming possible., Competing Interests: The authors have declared that no competing interests exist.

The migratory plant-parasitic nematode Bursaphelenchus xylophilus is the pathogen of the pine wilt disease (PWD), causing serious damage to pine forests in China. During the process of plant resistance to multiple pathogens, plant immunity plays a key role. In this current study, the pathogen-associated molecular pattern (PAMP) BxCDP1 in B. xylophilus has been identified, but the host target protein of BxCDP1 and its key amino acid region inducing the plant immunity have yet to be elucidated. We found that BxCDP1 could trigger superoxide production, H 2 O 2 production, and callose deposits. A RING-H2 finger protein 1 (RHF1) of Pinus thunbergii was screened and characterized as a target protein of BxCDP1 by yeast two-hybrid and co-immunoprecipitation (Co-IP). Moreover, two peptides (namely M9 and M16) proved to be key regions of BxCDP1 to induce PAMP-triggered immunity (PTI) in Nicotiana benthamiana , which also induced the expression of pathogenesis-related (PR) genes ( PtPR-3 , PtPR-4 , and PtPR-5 ) in P. thunbergii and enhanced the resistance of the host to B. xylophilus . These results indicate that BxCDP1 plays a critical role in the interaction between B. xylophilus and P. thunbergii , and both peptides M9 and M16 have the potential to be developed and utilized as immune inducers of pines against B. xylophilus in future., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hu, Wu, Wen, Ye, Qiu, Rui and Zhang.)

Bursaphelenchus xylophilus is the most economically important species of migratory plant-parasitic nematodes (PPNs) and causes severe damage to forestry in China. The successful infection of B. xylophilus relies on the secretion of a repertoire of effector proteins. The effectors, which suppress the host pine immune response, are key to the facilitation of B. xylophilus parasitism. An exhaustive list of candidate effectors of B. xylophilus was predicted, but not all have been identified and characterized. Here, an effector, named BxSCD3, has been implicated in the suppression of host immunity. BxSCD3 could suppress pathogen-associated molecular patterns (PAMPs) PsXEG1- and INF1-triggered cell death when it was secreted into the intracellular space in Nicotiana benthamiana. BxSCD3 was highly up-regulated in the early infection stages of B. xylophilus . BxSCD3 does not affect B. xylophilus reproduction, either at the mycophagous stage or the phytophagous stage, but it contributes to the virulence of B. xylophilus. Moreover, BxSCD3 significantly influenced the relative expression levels of defense-related (PR) genes PtPR-3 and PtPR-6 in P inus thunbergii in the early infection stage. These results suggest that BxSCD3 is an important toxic factor and plays a key role in the interaction between B. xylophilus and host pine.

A previous study found that a biocontrol bacterium, Bacillus pumilus HR10, inhibited the Sphaeropsis shoot blight disease of pine, and the fermentation broth of HR10 strain contained protein antifungal substances. The optimal formulation of the fermentation medium for the antagonistic substance of B. pumilus HR10 was finally obtained by single-factor test, Packett-Burman test, steepest ascent test and Box-Behnken Design (BBD) response surface test, and the best formulation of the fermentation medium for the antagonistic substance of B. pumilus HR10 was 12 g/L corn meal, 15 g/L beef extract and 13 g/L magnesium sulfate, with a predicted bacterial inhibition rate of 89%. The fermentation filtrate of B. pumilus HR10 cultured with the optimized medium formulation was verified to have an inhibition rate of (87.04 ± 3.2) % on the growth of Sphaeropsis sapinea by three replicate tests. The antagonistic crude protein of B. pumilus HR10 were further isolated and identified using HiTrap Capto Q strong Ion-Exchange Chromatography and LC-MS-MS, and it was speculated that glycoside hydrolase (Ghy), beta-glucanase (Beta), arabinogalactan endonuclease β-1,4-galactanase (Arab), and immunosuppressant A (ImA) are proteins with antagonistic activity against S. sapinea in the B. pumilus HR10., (© 2022. The Author(s).)

Background: The pine wilt disease (PWD) caused by Bursaphelenchus xylophilus is a devastating forest disease and its pathogenesis remains unclear. Secreted enzymes and proteins are important pathogenicity determinants and Bx-FAR-1 is an important pathogenic protein involved in the interaction between pine and B. xylophilus. However, the function of the Bx-FAR-1 protein in monitoring and prevention PWD remains unknown., Results: We found a small peptide of B. xylophilus effector Bx-FAR-1 is sufficient for immunosuppression function in Nicotiana benthamiana. Transient expression of Bx-FAR-1 in N. benthamiana revealed that nuclear localization is required for its function. The results of the ligand binding test showed that Bx-FAR-1 protein had the ability to bind fatty acid and retinol. We demonstrated that Bx-FAR-1 targeted to the nuclei of Pinus thunbergii using the polyclonal antibody by immunologic approach. The content of jasmonic acid (JA) was significantly increased in P. thunbergii infected with B. xylophilus when Bx-FAR-1 was silenced. We identified an F-box protein as the host target of Bx-FAR-1 by yeast two-hybrid and co-immunoprecipitation. Moreover, we found that Pt-F-box-1 was up-regulated during B. xylophilus infection and the expression of Pt-F-box-1 was increased in Bx-FAR-1 double-stranded RNA (dsRNA)-treated host pines., Conclusion: This study illustrated that Bx-FAR-1 might mediate the JA pathway to destroy the immune system of P. thunbergii, indicating that PWN likely secretes effectors to facilitate parasitism and promote infection, which could better reveal the pathogenesis mechanisms of B. xylophilus and would be beneficial for developing disease control strategies., (© 2022 Society of Chemical Industry.)

Background: Bursaphelenchus xylophilus is the causal agent of pine wilt disease (PWD) that has caused enormous ecological and economic losses in China. The mechanism in the interaction between nematodes and pine remains unclear. Plant parasitic nematodes (PPNs) secrete effectors into host plant tissues. However, it is poorly studied that role of effector in the infection of pine wood nematode (PWN)., Results: We cloned, characterized and functionally validated the B. xylophilus effector BxML1, containing an MD-2-related lipid-recognition (ML) domain. This protein inhibits immune responses triggered by the molecular pattern BxCDP1 of B. xylophilus. An insitu hybridization assay demonstrated that BxML1 was expressed mainly in the dorsal glands and intestine of B. xylophilus. Subcellular localization analysis showed the presence of BxML1 in the cytoplasm and nucleus. Furthermore, number of B. xylophilus and morbidity of pine were significantly reduced in Pinus thunbergii infected with B. xylophilus when BxML was silenced. Using yeast two-hybrid (Y2H) and coimmunoprecipitation (CoIP) assays, we found that the BxML1 interacts with cyclophilin protein PtCyP1 in P. thunbergii., Conclusions: This study illustrated that BxML1 plays a critical role in the B. xylophilus-plant interaction and virulence of B. xylophilus., (© 2022. The Author(s).)

The plant-parasitic nematode Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), causes enormous economic loss every year. Currently, little is known about the pathogenic mechanisms of PWD. Several effectors have been identified in B. xylophilus, but their functions and host targets have yet to be elucidated. Here, we demonstrated that BxSCD1 suppresses cell death and inhibits B. xylophilus PAMP BxCDP1-triggered immunity in Nicotiana benthamiana and Pinus thunbergii. BxSCD1 was transcriptionally upregulated in the early stage of B. xylophilus infection. In situ hybridization experiments showed that BxSCD1 was specifically expressed in the dorsal glands and intestine. Cysteine residues are essential for the function of BxSCD1. Transient expression of BxSCD1 in N. benthamiana revealed that it was primarily targeted to the cytoplasm and nucleus. The morbidity was significantly reduced in P. thunbergii infected with B. xylophilus when BxSCD1 was silenced. We identified 1-aminocyclopropane-1-carboxylate oxidase 1, the actual ethylene-forming enzyme, as a host target of BxSCD1 by yeast two-hybrid and coimmunoprecipitation. Overall, this study illustrated that BxSCD1 played a critical role in the B. xylophilus-plant interaction., (© 2021 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

The blood-cerebrospinal fluid barrier (BCSFB) is another gatekeeper between systemic circulation and the central nervous system (CNS), mainly present at the boundary between choroid plexuses and the ventricular system. This study demonstrates BCSFB opening in rats by single pulse of low-energy focused shockwave (FSW, energy flux density 0.03 mJ/mm 2 , 2 × 10 6 microbubbles/kg) treatment at lateral ventricle, resulting in significantly elevated cerebrospinal fluid (CSF) concentrations of systemically-administered gastrodin (GTD) (4 times vs. control within 3 hrs) that remained detectable for 24 hrs. The FSW-GTD group had significantly lower Racine's scale (<4) and zero mortality (n = 30) after lithium-pilocarpine-induced epilepsy. Electrophysiological recordings showed decreased epileptiform discharges, and brain section histology revealed reduced inflammation, oxidative stress and apoptosis, when compared with groups without FSW (Racine's scale: 4 ∼ 5; mortality: 26.67 ∼ 36.67%). FSW-mediated BCSFB opening provides a promising alternative for controlled-delivery of therapeutics into the CNS, offering rapid and widespread medication distribution. The technique could by applied in the development of novel therapies for various CNS diseases., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

The thermal management of microelectronics is important because overheating can lead to various reliability issues. The most common thermal solution used in microelectronics is forced convection, which is usually initiated and sustained by an airflow generator, such as rotary fans. However, traditional rotary fans might not be appropriate for microelectronics due to the space limit. The form factor of an ionic wind pump can be small and, thus, could play a role in the thermal management of microelectronics. This paper presents how the performance of a needle-ring ionic wind pump responds to inlet blockage in different electrical driving modes (direct current), including the flow rate, the corona power, and the energy efficiency. The results show that the performance of small needle-ring ionic wind pumps is sensitive to neither the inlet blockage nor the electrical driving mode, making needle-ring ionic wind pumps a viable option for microelectronics. On the other hand, it is preferable to drive needle-ring ionic wind pumps by a constant current if consistent performance is desired.

To evaluate the efficacy and safety of Compound Glycyrrhizin Injection(CGI) in improving liver damage in chronic hepatitis B(CHB). PubMed, Web of Science, SinoMed, CNKI, Wanfang and VIP databases were retrieved from their inception to February 10, 2020. The randomized controlled trial(RCT) of CGI in the treatment of CHB was included. Data were independently extracted by two authors, and the methodological quality was evaluated using the Cochrane bias risk assessment tool by other two authors. Statistical analysis was performed using RevMan 5.3 software. A total of 18 two-armed RCTs were included, involving 1 915 participants. The methodological quality of all studies included was generally low. In the comparison between CGI and diammonium glycyrrhizinate, the results showed that CGI was superior to the control group in improving the overall clinical effectiveness, but there was no statistical difference between the two groups in increasing ALT normalization rate, reducing ALT and AST level. In the comparison between CGI and diammonium glycyrrhizinate+other general hepatoprotective drugs, the results showed that CGI was superior to the control group in reducing AST level, while there was no statistical difference between the two groups in reducing ALT level and increasing overall clinical effectiveness. In the comparison between CGI+other commonly used drugs(including energy mixture, glutathione, vitamins, potassium magnesium aspartate) and diammonium glycyrrhizinate+other commonly used drugs, the results showed that CGI combined with other commonly used drugs was better than the control group in reducing ALT and AST level and improving the clinical total effective rate, and there was no statistical difference between the two groups in increasing the rate of ALT normalization. In the comparison between CGI+other commonly used drugs and other commonly used drugs, the results showed that CGI combined with other commonly used drugs was superior to the control group in reducing ALT and AST level and improving the overall clinical effectiveness. In the comparison between CGI+vitamins and diammonium glycyrrhizinate+potassium magnesium aspartate+vitamins, the results showed no statistical difference between the two groups in reducing AST level. A small number of studies included reported that CGI caused mild adverse reactions when used alone or in combination with other drugs. Based on the results, CGI has a certain effect in improving CHB liver damage, but the evidence is not enough to prove that CGI would cause serious adverse events. In the future, more well-designed and strictly-enforced RCT with an adequate sample size are needed to further evaluate the effect CGI in alleviating CHB liver damage.

Objective: To investigate the clinical features, survival and prognostic factors of elder patients with diffuse large B-cell lymphoma (DLBCL). Methods: The clinical data of elder patients with diffuse large B-cell lymphoma enrolled in the Cancer Hospital of Chinese Academy of Medical Sciences from April 2006 to December 2012 were retrospectively collected. All the patients were divided into R-CHOP-like group and CHOP-like group according to the dosage regimen. And the differences in demographic characteristics, clinical features, survival time and prognostic factors were compared between these two groups. Results: A total of 158 patients were enrolled, of which 78 patients in the R-CHOP-like group and 80 patients in the CHOP-like group were eligible. There were no significant differences between two groups on age, gender, pathological staging, B symptoms, bulky mass, ECOG score, IPI score, pathological type, LDH level, β(2)-MG level, lymphocyte/monocyte ratio(LMR), neutrophils/lymphocyte ratio(NLR), platelet/lymphocyte ratio(PLR), Ki-67 index and bone marrow invasion. In the R-CHOP like group, the median progression-free survival (PFS) time was 10 months, and the median overall survival (OS) time was 30 months. The 1-year and 2-year PFS rates were 46.2% and 19.2%, respectively. The 1-, 2-, and 5-year OS rates were 79.5%, 59.0%, and 19.2%, respectively. In the CHOP-like group, the median PFS was 7 months, and the median OS was 15 months. The 1-year and 2-year PFS rates were 27.5% and 12.5% respectively. The 1-year, 2-year, and 5-year OS rates were 65.0%, 32.5% and 13.8%, respectively. The median PFS time and OS time in the R-CHOP group were significantly better than those in the CHOP group ( P <0.05 for both). A stratified analysis showed that the PFS time and OS time were superior in the R-CHOP-like group compared to the CHOP-like group among patients older than 70 years ( P <0.05 for both). In patients with stage Ⅲ-Ⅳ, the PFS time and OS time in the R-CHOP-like group were also superior to CHOP-like group ( P <0.05 for both). Univariate Cox regression analysis showed that IPI score, LDH value, β(2)-MG value, ECOG score, LMR, and PLR had an significant effect on prognosis ( P <0.05 for all). Multivariate Cox regression analysis showed that lymphocyte/monocyte ratio and platelet/lymphocyte ratio were independent prognostic factors for diffuse large B-cell lymphoma ( P <0.05 for both). Conclusions: The R-CHOP-like chemotherapy regimen is superior to the CHOP-like regimen in the first-line treatment of patients with diffuse large B-cell lymphoma. ECOG score, LMR and PLR may be independent prognostic factors for diffuse large B-cell lymphoma. ECOG score, LMR and PLR are independent prognostic factors.

Electrostatic precipitator (ESP) is an electrohydrodynamic-based air filter that charges particles based on corona discharge and collects particles by induced electrostatic forces. Inducing corona discharge requires strong electric fields that, however, bring reliability issues because of oxidation. This paper presents the characteristics of an ESP that uses the cylindrical corona electrodes whose longitudinal axis is perpendicular to the surface of the ground electrode. The characteristics include the current-voltage curve, the surface oxidation of the cylindrical corona electrodes, and the element analysis. The characteristics are presented with respect to the pitch and diameter of the cylindrical corona electrodes. The results show that the characteristics mentioned above can correlate to the electric fields around the cylindrical corona electrodes. Stronger electric field around the cylindrical corona electrode results in higher collection efficiency, more oxidation on the cylindrical corona electrode, and shorter life of the cylindrical corona electrode., (© 2020 The Authors.)

Activated carbon is the most known material used to adsorb ozone. Activating carbonaceous materials by ozonation is commonly used to produce activated carbon, however, requiring sophisticated skills and professional equipment. This paper presents a reversed idea: to adsorb ozone using an unactivated carbonaceous material, coffee. Three powder adsorbents are presented: fresh coffee (unactivated), spent coffee grounds (unactivated), and activated carbon (commercially available). The test is conducted by measuring and comparing the ozone concentration in an ozone-supplied chamber with or without the ozone adsorbent. The results show that, at the specific conditions, the peak ozone concentration is lowered by 38% to 56% when the chamber has the activated carbon. At the same conditions, the peak ozone concentration is lowered by 25% to 43% when the chamber has the coffee powders (either fresh or spent). The elemental analysis demonstrates that the oxygen content after the ozone adsorption increases by 20%, 14.4%, and 34.5% for the fresh coffee, the spent coffee grounds, and the activated carbon, respectively. The characteristic analysis (the Fourier-transform infrared spectroscopy, the thermogravimetric, and the Brunauer-Emmett-Teller) suggests that the unactivated coffee is not porous, however, contains various organic compounds that could react with and consume ozone.

Background: Elderly patients with diffuse large B-cell lymphoma (DLBCL) have a worse prognosis than younger patients, and the optimal treatment strategy for this group remains controversial. We conducted a retrospective analysis to investigate the clinical features and outcomes of elderly patients (>60 years) and to assess the impact of clinical and molecular factors on outcome in this age group., Methods: From April 2006 to December 2012, a total of 349 elderly patients with DLBCL from the National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences, and Peking Union Medical College were included in this analysis. Patients were further divided into two age groups (61-69 years and ≥70 years). We compared clinical characteristics and outcomes between groups., Results: Of 349 total patients, 204 (58.5%) were aged 61 to 69 years, and 145 (41.5%) patients were aged 70 years or older. Except for the Eastern Cooperative Oncology Group performance status, clinical characteristics were comparable between the two groups. With a median follow-up of 82 (range, 1-129) months, the 5-year overall survival (OS) and progression-free survival (PFS) rates were 51.9% and 45.8%, respectively. The 5-year OS rates for patients aged 61 to 69 years and those over 70 years were 58.3% and 42.8% (P = 0.007), respectively, and the 5-year PFS rates were 51.0% and 38.6% (P = 0.034). Treatment regimens including rituximab provided a higher 5-year OS rate (63.1% vs. 37.1%, P < 0.001) and PFS rate (56.6% vs. 31.8%, P < 0.001) than chemotherapy alone. For patients aged 61 to 69 years, chemotherapy plus rituximab resulted in a higher 5-year OS rate (66.7% vs. 46.4%, P = 0.002) and PFS rate (60.0% vs. 38.1%, P = 0.002) than chemotherapy alone. For patients aged ≥70 years, there was a marked survival advantage in patients who received chemotherapy plus rituximab (5-year OS rate: 57.7% vs. 25.4%, P < 0.001; 5-year PFS rate: 51.3% vs. 23.9%, P < 0.001) compared with that seen in those who received chemotherapy alone. Multivariate analysis established that stage III/IV disease, elevated lactate dehydrogenase (LDH), initial treatment, and chemotherapy with rituximab were independent risk factors for 5-year OS, and stage III/IV disease, elevated LDH, and chemotherapy with rituximab were independent risk factors for 5-year PFS for elderly patients with DLBCL., Conclusions: In comparison to patients aged 61 to 69 years, those aged ≥70 years have poorer survival. Prolonged survival is obtainable with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP)-like in elderly Chinese patients in all age groups, indicating that the R-CHOP-like regimen should be considered for this population, even for those aged 70 years or older.

Epithelial cell transforming sequence 2 (ECT2) is a guanine nucleotide exchange factor and its expression is associated with the development of malignant tumor types. However, to the best of our knowledge, there is no information on the role of ECT2 in the development and progression of laryngeal squamous cell carcinoma (LSCC). The present study aimed at investigating the expression pattern and potential role of ECT2 in the development and progression of LSCC. The expression of ECT2 in 81 pairs of LSCC and adjacent non-tumor tissues was characterized by immunohistochemistry and reverse transcription-quantitative polymerase chain reaction. ECT2 expression was upregulated in LSCC tissues and associated significantly with poor differentiation, higher stages, lymph node metastasis and poor survival in the sample population. The relative expression levels of ECT2 mRNA transcripts were correlated with the intensity of anti-ECT2 staining in 25 ECT2+LSCC specimens selected randomly. These results indicated that ECT2 expression was crucial for the progression of LSCC and may serve as a biomarker for the diagnosis and prognosis of LSCC.

Background: Intervertebral disc (IVD) degeneration is a condition characterized by a reduction in the water and extracellular matrix content of the nucleus pulposus (NP) and is considered as one of the dominating contributing factors to low back pain. Recent evidence suggests that stromal cell-derived factor 1α (SDF-1α) and its receptor C-X-C chemokine receptor type 4 (CXCR4) direct the migration of stem cells associated with injury repair in different musculoskeletal tissues., Aim: To investigate the effects of SDF-1α on recruitment and chondrogenic differentiation of nucleus pulposus-derived stem cells (NPSCs)., Methods: We performed real-time RT-PCR and enzyme-linked immunosorbent assay to examine the expression of SDF-1α in nucleus pulposus cells after treatment with pro-inflammatory cytokines in vitro . An animal model of IVD degeneration was established using annular fibrosus puncture in rat coccygeal discs. Tissue samples were collected from normal control and degeneration groups. Differences in the expression of SDF-1α between the normal and degenerative IVDs were analyzed by immunohistochemistry. The migration capacity of NPSCs induced by SDF-1α was evaluated using wound healing and transwell migration assays. To determine the effect of SDF-1α on chondrogenic differentiation of NPSCs, we conducted cell micromass culture and examined the expression levels of Sox-9, aggrecan, and collagen II. Moreover, the roles of SDF-1/CXCR4 axis in the migration and chondrogenesis differentiation of NPSCs were analyzed by immunofluorescence, immunoblotting, and real-time RT-PCR., Results: SDF-1α was significantly upregulated in the native IVD cells cultured in vitro with pro-inflammatory cytokines, such as interleukin-1β and tumor necrosis factor-α, mimicking the degenerative settings. Immunohistochemical staining showed that the level of SDF-1α was also significantly higher in the degenerative group than in the normal group. SDF-1α enhanced the migration capacity of NPSCs in a dose-dependent manner. In addition, SDF-1α induced chondrogenic differentiation of NPSCs, as evidenced by the increased expression of chondrogenic markers using histological and immunoblotting analyses. Real-time RT-PCR, immunoblotting, and immunofluorescence showed that SDF-1α not only increased CXCR4 expression but also stimulated translocation of CXCR4 from the cytoplasm to membrane, accompanied by cytoskeletal rearrangement. Furthermore, blocking CXCR4 with AMD3100 effectively suppressed the SDF-1α-induced migration and differentiation capacities of NPSCs., Conclusion: These findings demonstrate that SDF-1α has the potential to enhance recruitment and chondrogenic differentiation of NPSCs via SDF-1/CXCR4 chemotaxis signals that contribute to IVD regeneration., Competing Interests: Conflict-of-interest statement: To the best of our knowledge, no conflict of interest exists.

A gradient heterosturcture is one of the basic methods to control the charge flow in perovskite solar cells (PSCs). However, a classical route for gradient heterosturctures is based on the diffusion technique, in which the guest ions gradually diffuse into the films from a concentrated source of dopants. The gradient heterosturcture is only accessible to the top side, and may be time consuming and costly. Here, the "intolerant" n-type heteroatoms (Sb 3+ , In 3+ ) with mismatched cation sizes and charge states can spontaneously enrich two sides of perovskite thin films. The dopants at specific sides can be extracted by a typical hole-transport layer. Theoretical calculations and experimental observations both indicate that the optimized charge management can be attributed to the tailored band structure and interfacial electronic hybridization, which promote charge separation and collection. The strategy enables the fabrication of PSCs with a spontaneous graded heterojunction showing high efficiency. A champion device based on Sb 3+ doped film shows a stabilized power-conversion efficiency of 21.04% with a high fill factor of 0.84 and small hysteresis., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

As one of the most promising light-harvesting materials, perovskites have drawn tremendous attention for their unique advantages, such as high efficiency, low cost and facile fabrication compared with other photovoltaic materials. Nevertheless, poor moisture tolerance of the perovskites greatly hampers the operation of such devices and hinders their commercialization. Herein, we demonstrate a facile dipping treatment using sulfur-oleylamine solution for surface atomic modulation of perovskite films. Oleylammonium polysulfides (OPs) would be self-assembled on the etched perovskite film as an ultrathin outer layer. This layer could passivate the surface chemical activity of the outer perovskite layers. Moreover, the hydrophobic OPs significantly enhance moisture stability of such devices. As a result, the obtained device without encapsulation retains more than 70% of its initial power conversion efficiency (PCE) after 14 days of exposure to a relative humidity of 40 ± 10%.

A highly transparent NiO layer was prepared by a solution processing method with nickel(II) 2-ethylhexanoate in non-polar solvent and utilized as HTM in perovskite solar cells. Excellent optical transmittance and the matched energy level lead to the enhanced power conversion efficiency (PCE, 18.15 %) than that of conventional sol-gel-processed NiO-based device (12.98 %)., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: To establish Surface-enhanced Raman Spectroscopy (SERS) can be used as a rapid and reliable method to distinguish virulent strain and mild strain of L. pneumophila., Methods: Mortality data were collected from company departments through administrative documents, death certificates, etc. Trend analyses of cancer mortality were performed on the basis of 925 cancer deaths between 2001 and 2010., Results: Our results indicated that the peaks of high virulence strains reached ⋝4000. This criterion was verified by subsequent cell experiments. In addition, we also conducted SERS rapid identification on the virulence of several collected clinical strains and obtained accurate results., Conclusion: The present study indicates that the established SERS protocol can be used as a rapid and reliable method to distinguish virulent and mildly virulent strains of L. pneumophila, which can be further used in clinical samples., (Copyright © 2015 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

Nucleus pulposus (NP) tissue engineering has been proposed as a novel biological treatment for early-stage intervertebral disc degeneration. In this study, a novel functional self-assembling peptide PKP was first designed by linking the short functional motif of bone morphogenetic protein-7 (BMP7) to the C-terminal of RADA16-I, and another new functional self-assembling peptide was obtained by mixing RKP with RADA16-I. Then, the biocompatibilities and bioactivities of RKP and RAD-RKP for human degenerated nucleus pulposus cells (hNPCs) were studied in vitro. Atomic force microscopy and scanning electron microscopy (SEM) confirmed that both RKP and RAD-RKP could self-assemble into three-dimensional (3D) nanofiber hydrogel scaffolds in a culture medium at 37°C. After the hNPCs were cultured in 3D scaffolds, both RKP and RAD-RKP exhibited reliable attachment and extremely low cytotoxicities (<14%), which were verified by SEM and cytotoxity assays, respectively. Our results also showed that the functional-based scaffolds could increase the proliferation and migration of hNPCs after 7 days compared with culture plates and pure RADA16-I. Quantitative real-time polymerase chain reaction demonstrated that the expressions of collagen II α1, Sox-9, and aggrecan were upregulated, while collagen I α1 was downregulated by functional-based scaffolds after 28 days. Furthermore, we also confirmed that RAD-RKP exhibited a higher hNPC proliferation, migration, and expression of Sox-9 and aggrecan compared with pure RKP. Therefore, the results of this study indicated that the BMP7 short motif-designed functional self-assembling peptide nanofiber hydrogels could be used as excellent scaffolds in NP tissue engineering, and RAD-RKP might have further potential application in human mild degenerated NP tissue regeneration.

PDZ domain-containing proteases, also known as HtrA family proteases, play important roles in bacterial cells by modulating disease pathogenesis and cell-envelope stress responses. These proteases have diverse functions through proteolysis- and nonproteolysis-dependent modes. Here, we report that the genome of the causative agent of rice bacterial blight, Xanthomonas oryzae pv. oryzae, encodes seven PDZ domain-containing proteins. Systematic inactivation of their encoding genes revealed that PXO&#95;01122 and PXO&#95;04290 (prc) are involved in virulence. prc encodes a putative HtrA family protease that localizes in the bacterial periplasm. Mutation of prc also resulted in susceptibility to multiple environmental stresses, including H2O2, sodium dodecylsulfate, and osmolarity stresses. Comparative subproteomic analyses showed that the amounts of 34 periplasmic proteins were lower in the prc mutant than in wild-type. These proteins were associated with proteolysis, biosynthesis of macromolecules, carbohydrate or energy metabolism, signal transduction, and protein translocation or folding. We provide in vivo and in vitro evidence demonstrating that Prc stabilizes and directly binds to one of these proteins, DppP, a dipeptidyl peptidase contributing to full virulence. Taken together, our results suggest that Prc contributes to bacterial virulence by acting as a periplasmic modulator of cell-envelope stress responses.

A novel antioxidant polysaccharide-peptide complex LB-1b from the fruiting bodies of the edible abalone mushroom (Pleurotus abalonus) was purified and identified. The structural characteristic of LB-1b was identified by FTIR (Fourier-transform IR), 13C NMR and 1H NMR spectroscopy. LB-1b is a polysaccharide-peptide complex that contains glucose, rhamnose, glucuronic acid and galactose in the molar ratio of 22.4:1:1.7:1.6 and the N-terminal sequence of its peptide moiety has also been determined. The N-terminal amino acid sequence of LB-1b, IPKERKEFQQAQHLK, showed some resemblance to antioxidant enzymes. LB-1b exhibited high antioxidant activity in erythrocyte haemolysis in vitro and the anti-proliferative activity towards hepatoma HepG2 cells and breast cancer MCF7 cells with an IC50 of 24 and 14 μM respectively. LB-1b also demonstrated hypoglycaemic activity in drug-induced diabetic mice and anti-HIV-1 RT (reverse transcriptase) with an IC50 value of 12.5 μM.

Objective: The intent of this study was to purify and characterize a polysaccharide named LA from the fruiting bodies of the edible mushroom Pleurotus abalones., Methods: The 120-kDa polysaccharide was obtained by extraction with boiling water, ethanol precipitation, ion exchange chromatography on Diethylaminoethyl-cellulose and gel filtration on Sephadex G-200., Key Findings: The LA was composed of glucose, rhamnose, glucuronic acid, xylose, galactose and arabinose in the molar ratio of 26.3:2.7:1:1.4:1.8:1.2. The FITR and 1H-NMR spectrum of LA disclosed that it was a saccharide with an α-configuration. Its 13C-NMR spectrum revealed that its main chain was [→6)-α-D-Gly(1→]n. The LA exhibited antioxidant activities, especially in scavenging 1,1- diphenyl-2-picryl-hydrazyl radicals and hydroxyl radicals. It manifested antiproliferative activity towards breast cancer MCF7 cells with an IC50 of 3.7 µm, and also exerted some antiproliferative activity against HepG2 cells. It manifested a hypoglycemic action on diabetic mice. It inhibited HIV-1 reverse transcriptase with an IC50 of 8.7 × 10(-2) µm., Conclusions: The polysaccharide from the abalone mushroom., (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)

Bacillus amyloliquefaciens has been a major workhorse for the production of a variety of commercially important enzymes and metabolites for the past decades. Some subspecies of this bacterium are recalcitrant to exogenous DNA, and transformation with plasmid DNA is usually less efficient, thereby limiting the genetic manipulation of the recalcitrant species. In this work, a methodology based on electro-transformation has been developed, in which the cells were grown in a semicomplex hypertonic medium, cell walls were weakened by adding glycine (Gly) and DL-threonine (DL-Thr), and the cell-membrane fluidity was elevated by supplementing Tween 80. After optimization of the cell-loosening recipe by response surface methodology (RSM), the transformation efficiency reached 1.13 ± 0.34 × 10(7) cfu/μg syngeneic pUB110 DNA in a low conductivity electroporation buffer. Moreover, by temporary heat inactivation of the host restriction enzyme, a transformation efficiency of 8.94 ± 0.77 × 10(5) cfu/μg DNA was achieved with xenogeneic shuttle plasmids, a 10(3)-fold increase compared to that reported previously. The optimized protocol was also applicable to other recalcitrant B. amyloliquefaciens strains used in this study. This work could shed light on the functional genomics and subsequent strain improvement of the recalcitrant Bacillus, which are difficult to be transformed using conventional methods., (Copyright © 2010 Elsevier Inc. All rights reserved.)

According to previous studies, a close relationship between oxidative stress and AIDS suggests that antioxidants might play an important role in the treatment of AIDS. Cordyceps militaris was selected from nine edible mushrooms by assay of inhibition of erythrocyte hemolysis. Macroporous adsorption resin and HPLC were used to purify three micromolecular compounds named L3a, L3b and L3c. L3a was identified to be adenosine with the molecular formula C(10)H(13)N(5)O(4); L3b was 6,7,2',4',5'-pentamethoxyflavone with the molecular formula C(20)H(20)O(7), and L3c was dimethylguanosine with the molecular formula C(12)H(17)N(5)O(5). The compound 6,7,2',4',5'-pentamethoxyflavone was first isolated from C. militaris. The assay of inhibition of HIV-1 protease (HIV-1 PR) was based on the fact that the expression of this enzyme can inhibit the growth of E. coli. This is a new screening system for HIV-1 PR inhibitors. Both L3a and L3b showed high inhibition to HIV-1 PR. These compounds could be new anti-HIV-1 PR drugs., (2010 Elsevier GmbH. All rights reserved.)

Objective: We compared the anesthetic efficacy of inferior alveolar nerve block (IANB) plus buccal infiltration (BI) and IANB plus periodontal ligament (PDL) articaine injections in patients with irreversible pulpitis in the mandibular first molar., Study Design: Fifty-seven volunteers, patients with irreversible pulpitis in the mandibular first molar admitted to the Department of Stomatology, Second Affiliated Hospital, Sun Yat-Sen University, randomly received conventional IANB, containing 1.7 mL 4% articaine/HCl with 1:100,000 epinephrine, plus either BI or PDL injections containing 0.4 mL articaine/HCl with 1:100,000 epinephrine. The patients recorded the pain of the injections and endodontic access on a Heft-Parker visual analog scale (VAS)., Results: According to the VAS scores, all patients experienced no or mild pain with BI and PDL injections after the application of IANB. Anesthetic success occurred in 81.48% for IANB plus BI (IANB/BI) compared with 83.33% for IANB plus PDL injection (IANB/PDL injection). None of the observed differences between the 2 groups was significant (P > .05)., Conclusion: Both injection combinations resulted in high anesthetic success in patients with irreversible pulpitis in the mandibular first molar.

In this study, the fed-batch fermentation technique was applied to improve the yield of L-threonine produced by Escherichia coli TRFC. Various fermentation substrates and conditions were investigated to identify the optimal carbon source, its concentration and C/N ratio in the production of L-threonine. Sucrose was found to be the optimal initial carbon source and its optimal concentration was determined to be 70 g/L based on the results of fermentations conducted in a 5-L jar fermentor using a series of fed-batch cultures of E. coli TRFC. The effects of glucose concentration and three different feeding methods on the production of L-threonine were also investigated in this work. Our results showed that the production of L-threonine by E. coli was enhanced when glucose concentration varied between 5 and 20 g/L with DO-control pulse fed-batch method. Furthermore, the C/N ratio was a more predominant factor than nitrogen concentration for L-threonine overproduction and the optimal ratio of ammonium sulfate to sucrose (g/g) was 30. Under the optimal conditions, a final L-threonine concentration of 118 g/L was achieved after 38 h with the productivity of 3.1 g/L/h (46% conversion ratio from glucose to threonine).

The cyclic lipopeptide (CLP) has been known to inhibit proliferation and induce apoptosis in cancer cells. However, the molecular mechanisms involved in CLP-induced apoptosis are still uncharacterized in human leukemic K562 cells. The current study investigated the molecular mechanism of action of CLP, purified from Bacillus natto T-2. CLP-induced a sustained increase in concentration of intracellular Ca(2+). This increase in [Ca(2+)]i was associated with CLP-induced cell apoptosis and ERK phosphorylation. CLP-induced cell apoptosis was reversed by PD98059 (an inhibitor of ERK), but not by SB203580 (an inhibitor of p38) and SP200125 (an inhibitor of JNK), suggesting that the action of CLP on K562 cells was via ERK, but not via p38 and JNK. On the other hand, pretreatment with Bapta-AM, a well-known calcium chelator, partially blocked CLP-induced apoptosis, indicating that the elevation of [Ca(2+)]i may play an important role in the apoptosis. Collectively, in K562 cells, CLP-induced an increase in [Ca(2+)]i which evoked ERK phosphorylation. This ERK phosphorylation subsequently activated Bax, cytochrome c and caspase-3 leading to apoptosis.

River water and wastewater treatment plant (WWTP) effluents from metropolitan Taipei, Taiwan were tested for the presence of the pollutants estrone (E1), estriol (E3), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2) using a new methodology that involves high-flow solid-phase extraction and liquid chromatography/tandem mass spectrometry. The method was also used to investigate the removal of the analytes by conventional drinking water treatment processes. Without adjusting the pH, we extracted 1-L samples with PolarPlus C18 Speedisks under a flow rate exceeding 100 mL/min, in which six samples could be done simultaneously using an extraction station. The adsorbent was washed with 40% methanol/60% water and then eluted by 50% methanol/50% dichloromethane. The eluate was concentrated until almost dry and was reconstituted by 20 microL of methanol. Quantitation was done by LC-MS/MS-negative electrospray ionization in the selected reaction monitoring mode with isotope-dilution techniques. The mobile phase was 10 mM N-methylmorpholine aqueous solution/acetonitrile with gradient elution. Mean recoveries of spiked Milli-Q water were 65-79% and precisions were within 2-20% of the tested concentrations (5.0-200 ng/L). The method was validated with spiked upstream river water; precisions were most within 10% of the tested concentrations (10-100 ng/L) with most RSDs<10%. LODs of the environmental matrixes were 0.78-7.65 ng/L. A pre-filtration step before solid-phase extraction may significantly influence the measurement of E1 and EE2 concentrations; disk overloading by water matrix may also impact analyte recoveries along with ion suppression. In the Taipei water study, the four steroid estrogens were detected in river samples (ca. 15 ng/L for E2 and EE2 and 35-45 ng/L for E1 and E3). Average levels of 19-26 ng/L for E1, E2, and EE2 were detected in most wastewater effluents, while only a single effluent sample contained E3. The higher level in the river was likely caused by the discharge of untreated human and farming waste into the water. In the drinking water treatment simulations, coagulation removed 20-50% of the estrogens. An increased dose of aluminum sulfate did not improve the performance. Despite the reactive phenolic moiety in the analytes, the steroids were decreased only 20-44% of the initial concentrations in pre- or post-chlorination. Rapid filtration, with crushed anthracite playing a major role, took out more than 84% of the estrogens. Except for E3, the whole procedure successfully removed most of the estrogens even if the initial concentration reached levels as high as 500 ng/L.

Brevibacterium flavum is used for the production of a number of amino acids in the biotechnology industry. The yield of producing a metabolite is ultimately limited by the ability of the central metabolism and the desired biosynthesis pathway. Pathway analysis is a very useful tool for metabolic engineering, which can be applied to increase the yield of a metabolite or channeling a metabolite into desired pathways. It does not require any kinetic parameters and only uses the Stoichiometric equations. Pathway analysis for production of L-leucine by Brevibacterium flavum TK0303 at steady state was conducted in this paper. Theoretical yield and flux distribution for optimal pathway were determined. It is also concluded that pyruvate and acetyl-coenzyme A are the key nodes of the L-leucine biosynthesis pathway by analyzing the flux distributions of different modes. According to the pathway analysis, the production of L-leucine is expected to be raised by strengthening the flux of the key nodes (pyruvate and acetyl-coenzyme A) through changing the environmental factors. Because the flux of TCA cycle in Brevibacterium flavum TK0303 is weak, the production of L-leucine must be provided enough amido by adding glutamic acid to the fermentation medium. NH4Ac is both a carbon source and a nitrogen source, which could be helpful to the production of L-leucine. The effects of glutamic acid and NH4Ac on the production of L-leucine were further studied. The production of L-leucine increased 56% by adding glutamic acid. By improving the concentration of NH4Ac, the biosynthesis of L-leucine was greatly strengthened too. The results indicate that the flux of L-leucine can be largely increased by changing the chemical regulatory factors such as NH4Ac and glutamic acid and the modes established by pathway analysis prove to be efficient to describe the metabolic network of L-leucine production by Brevibacterium flavum TK0303.

Failure of embryologic development of a lobe of the thyroid gland is a rare anomaly. We report a 47-year-old female with right thyroid hemiagenesis, papillary thyroid cancer, and an ectopic prelaryngeal thyroid rest. The preoperative diagnosis was made as a result of evaluating a progressively enlarging left thyroid mass by means of ultrasonography, technetium-99m thyroid scintigraphy, and fine-needle aspiration cytology. During the operation, an isthmus and an enlarged left lobe with a cancer mass 2 cm in diameter were found, and a solitary piece of thyroid tissue about 0.7 cm in length was present just below the hyoid bone. We conclude that recognition of this rare anomaly by means of both scintigraphy and ultrasonography is valuable for a complete thyroidectomy, and for avoiding unnecessary contralateral dissection and damage.

We present a case with extremely high serum gastrin induced by atrophic gastritis and Helicobacter pylori infection. The patient, a 95-year-old male, was diagnosed with idiopathic chronic diarrhea. During diagnostic work-up, his fasting serum gastrin was up to 2078 pg/mL. The secretin test was negative for gastrinoma. Octreotide scan showed no suspicious lesion except for diffuse faint uptake over the gastric antrum on the 48-hour-delay film. Gastric acidity test revealed achlorhydria. On histology examination, atrophic gastritis with Helicobacter pylori infection was found in the gastric body, but the antral mucus was normal with a slight increase in gastrin-secreting cells. To our knowledge, such extremely high serum gastrin induced by atrophic gastritis and Helicobacter pylori infection has never been reported before.

Taxadiene synthase plays an important role in taxol biosynthesis. RT-PCR was used for cloning taxadiene synthase cDNA fragment from the cells of T. cuspidata. The cDNA was cloned into vector pGEM and transformed to E. coli J M109. The cloned cDNA named pCBMZ was further confirmed by Southern blotting assay and was sequenced. The result showed that taxadiene synthase cDNA of Taxus cuspidata was highly homologous with that of Taxus brevifolia.