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ObjectiveThe present study explored the construction and application of a standardized postoperative pain-management procedure for patients with perianal abscess.MethodsTwo study groups (the observation group and the intervention group) were established retrospectively. The observation group comprised 46 patients with perianal abscess who enrolled in this study between June 2019 and June 2020. The intervention group comprised 48 patients who enrolled in the study between July 2020 and July 2021. All patients were enrolled using the convenience sampling method. A pain-management team was established, and standardized procedure management was implemented in the intervention group, while routine pain management was implemented in the observation group. Indices related to the patients' postoperative pain-control satisfaction and rehabilitation were compared between the two groups.ResultsThe patients' pain-control satisfaction, wound edema score, edema disappearance time, urinary retention, and defecation difficulty following intervention were better in the intervention group than in the observation group, and the differences were statistically significant (P ConclusionThe implementation of the standardized postoperative pain-management procedure in patients with perianal abscess can effectively improve the patient's level of pain and satisfaction and promote rehabilitation.

To observe the effect of acupuncture and moxibustion on the water content of stratum corneum (WCSC), expression of serum inflammatory factors and aquaporin 3 (AQPA total of 24 male albino guinea pigs were randomly divided into a blank group (Before the intervention, compared with the blank group, the EASI scores and TEWL were increased in the remaining groups (Acupuncture and moxibustion could improve the epidermal water metabolism and skin tissue morphology in guinea pigs with eczema of skin damp-heat accumulation. Its mechanism may be related to regulating inflammatory factors, up-regulating the expression of AQP

Arbutin, a natural polyphenol, inhibits OS cell viability, invasion and migration by down‐regulating MTHFD1L expression. miR‐338‐3p directly targets MTHFD1L. This regulatory action is associated with the inactivation of the AKT/mTOR signaling pathway., Arbutin, a glycoside extracted from the plant Arctostaphylos uva‐ursi, has been previously reported to possess antioxidant, anti‐inflammatory and anticancer effects. Here, we investigated whether arbutin affects the proliferation of the cells of the osteosarcoma (OS) cell lines MG‐63 and SW1353. Arbutin suppressed OS cell viability in a dose‐ and time‐dependent manner, as shown by Cell Counting Kit‐8 assay. Furthermore, arbutin exposure decreased the protein levels of MTHFD1L, CCND1 and phosphorylated‐protein kinase B (AKT)/phosphorylated‐mammalian target of rapamycin (mTOR). Potential upstream miRNAs of MTHFD1L were predicted using TargetScan, PICTAR5, miRanda and miRWalk. We performed luciferase activity assays to show that miR‐338‐3p directly targets and negatively regulates the expression of MTHFD1L. Knockdown of miR‐338‐3p promoted cell invasion, migration and proliferation in arbutin‐treated OS cells via MTHFD1L. In summary, our data suggest that arbutin inhibits OS cell proliferation, migration and invasion via miR‐338‐3p/MTHFD1L and by inactivating the AKT/mTOR pathway.

Background Radiotherapy (RT) is one of the main treatments for men with prostate cancer (PCa). Yet, to date, with numerous sophisticated nano-formulations as radiosensitizers have been synthesized with inspiring therapeutic effect both in vitro and in vivo, there still lacks the successful clinical translation of such nanosystems. Meanwhile, almost all the attention has been paid on the enhanced dose deposition effect by secondary electrons of nanomaterials with high atomic numbers (Z), despite that cell-cycle arrest, DNA damage and also reactive oxygen species (ROS) production are critical working mechanisms accounting for radiosensitization. Methods Herein, an ‘all-purpose’ nanostrategy based on dose deposition enhancement, cell cycle arrest and ROS production as prostate cancer radiosensitizer for potential clinical translation was proposed. The rather simple structure of docetaxel loaded Au nanoparticles (NPs) with prostate specific membrane antigen (PSMA) ligand conjugation have been successfully synthesized by a rather facile protocol. Results Enhanced cellular uptake achieved via selective internalization of the NPs by PCa cells with positive PSMA expression could guarantee the enhanced dose deposition. Moreover, the as-synthesized nanosystem could arrest cell cycle at G2/M phases, which would reduce the ability of DNA damage repair for more irradiation sensitive of the PCa cells. Meanwhile, G2/M phases arrest would further promote cascade retention and enrichment of the NPs within the cells. Furthermore, ROS generation and double strand breaks greatly promoted by the NPs under irradiation (IR) could also provide an underlying basis for effective radiosensitizers. Conclusions Investigations from in vitro and in vivo confirmed the as-synthesized NPs as an effective nano-radiosensitizer with ideal safety. More importantly, all the moieties within the present nanosystem have been approved by FDA for the purpose of PCa treatment, thus making the it highly attractive for clinical translation.

The aim of the present study was to investigate the role of microRNA-21 (miR-21) in regulating the classical WNT/β-catenin signaling pathway by targeting low-density lipoprotein-related receptor 6 (LRP6) in non-alcoholic fatty liver disease (NAFLD). For this purpose, we established a NAFLD model by feeding C57BL/6J mice a methionine-choline-deficient diet. Antagomir-21 was then injected via the tail vein, and the expression levels of WNT/β-catenin signaling pathway-related proteins, such as LRP6, glycogen synthase kinase-3β (GSK3β), p-β-catenin, β-catenin and the downstream protein, peroxisome proliferator-activated receptor γ (PPAR-γ), and lipid metabolism-related genes, including sterol regulatory element-binding transcription factor 1c (SREBP1c), fatty acid synthase (FAS), carnitine palmitoyl transferase 1α (CPT1α) and adenosine 5-monophosphate (AMP)-activated protein kinase α (AMPKα), were detected. The results revealed that in the NAFLD model, LRP6 expression was negatively associated with miR-21 expression. After antago-nizing the expression of miR-21, the protein level of LRP6 was increased. In addition, the WNT/β-catenin signaling pathway was activated, and lipid accumulation and inflammation were alleviated in the liver. However, the expression of PPAR-γ was not inhibited following the upregulation of the WNT signaling pathway. Taken together, the results of this study demonstrate that the inhibition of miR-21 expression can alleviate NAFLD by targeting LRP6 to activate the WNT/β-catenin signaling pathway.

Objective : To explore the histological feature of the cervical disc degeneration in patients with degenerative ossification (DO) and its potential mechanisms.Methods : A total of 96 surgical segments, from cervical disc degenerative disease patients with surgical treatment, were divided into ossification group (group O, n=46) and non-ossification group (group NO, n=50) based on preoperative radiological exams. Samples of disc tissues and osteophytes were harvested during the decompression operation. The hematoxylin-eosin staining, Masson trichrome staining and Safranin O-fast green staining were used to compare the histological differences between the two groups. And the distribution and content of transforming growth factor (TGF)-β1, p-Smad2 and p-Smad3 between the two groups were compared by a semi-quantitative immunohistochemistry (IHC) method.Results : For all the disc tissues, the content of disc cells and collagen fibers decreased gradually from the outer annulus fibrosus (OAF) to the central nucleus pulposus (NP). Compared with group NO, the number of disc cells in group O increased significantly. But for proteoglycan in the inner annulus fibrosus (IAF) and NP, the content in group O decreased significantly. IHC analysis showed that TGF-β1, p-Smad2, and p-Smad3 were detected in all tissues. For group O, the content of TGF-β1 in the OAF and NP was significantly higher than that in group NO. For p-Smad2 in IAF and p-Smad3 in OAF, the content in group O were significantly higher than group NO.Conclusion : Histologically, cervical disc degeneration in patients with DO is more severe than that without DO. Local higher content of TGF-β1, p-Smad2, and p-Smad3 are involved in the disc degeneration with DO. Further studies with multi-approach analyses are needed to better understand the role of TGF-β/Smads signaling pathway in the disc degeneration with DO.

The World Health Organization emphasized the importance of goggles and face shields for protection of medical personnel at the outbreak of the COVID-19 pandemic. Unsurprisingly, almost all countries suffered from a critical supply shortage of goggles and face shields, as well as many other types of personal protective equipment (PPE), for a long period, owing to the lack of key medical material supplies and the inefficiency of existing fabrication methods arising from the need to avoid crowds during the outbreak of COVID-19. In this paper, we propose a novel combined shield design for eye and face protection that can be rapidly fabricated using three-dimensional printing technology. The designed prototype eye-face shield is accessible to the general public, offering more possibilities for yield improvement in PPE during emergent infectious disease events such as COVID-19.

The purpose of this study is to use the newly developed truly mesh-free method by Ls-dyna, the smooth particle Galerkin (SPG) method, to simulate the material failure process caused by the extraction of pedicle screws from the trabecular bone. The experimental and theoretical results are compared to verify the effectiveness of the SPG method. In addition, the trabecular bone material of Johnson-Cook elastoplastic material is used to compare the convergence difference between SPG and traditional finite element method in this test. The results show that the failure curve simulated by the SPG method is more reasonable, and it is not only faster than the FEM method in terms of mesh convergence but also solves the defect that the traditional FEM cannot capture the peak force.

The World Health Organization emphasized the importance of goggles and face shields for protection of medical personnel at the outbreak of the COVID-19 pandemic. Unsurprisingly, almost all countries suffered from a critical supply shortage of goggles and face shields, as well as many other types of personal protective equipment (PPE), for a long period, owing to the lack of key medical material supplies and the inefficiency of existing fabrication methods arising from the need to avoid crowds during the outbreak of COVID-19. In this context, we propose a novel combined shield design for eye and face protection that can be rapidly fabricated using three-dimensional printing technology. The designed prototype eye–face shield is accessible to the general public , offering more possibilities for yield improvement in PPE during emergent infectious disease events such as COVID-19.

Whether to perform hybrid surgery (HS) in contrast to anterior cervical discectomy and fusion (ACDF) when treating patients with multilevel cervical disc degeneration remains a controversial subject. To resolve this we have undertaken a meta-analysis comparing the outcomes from HS with ACDF in this condition.Seven databases were searched for studies of HS and ACDF from inception of the study to 1 September 2019. Both random-effects and fixed-effects models were used to evaluate the overall effect of the C2-C7 range of motion (ROM), ROM of superior/inferior adjacent levels, adjacent segment degeneration (ASD), heterotopic ossification (HO), complications, neck disability index (NDI) score, visual analogue scale (VAS) score, Japanese Orthopaedic Association (JOA) score, Odom's criteria, blood loss, and operating and hospitalization time. To obtain more credible results contour-enhanced funnel plots, Egger's and Begg's tests, meta-regression, and sensitivity analyses were performed.In total, 17 studies involving 861 patients were included in the analysis. HS was found to be superior to ACDF in maintaining C2-C7 ROM and ROM of superior/inferior adjacent levels, but HS did not reduce the incidence of associated level ASD. Also, HS did not cause a higher rate of HO than ACDF. The frequency of complications was similar between the two techniques. HS failed to achieve more favourable outcomes than ACDF using the NDI, VAS, JOA, and Odom's scores. HS did not show any more advantages in operating or hospitalization time but did show reduction in blood loss.Although HS maintained cervical kinetics, it failed to reduce the incidence of ASD. This finding differs from previous reports. Moreover, patients did not show more benefits from HS with respect to symptom improvement, prevention of complications, and clinical outcomes. Cite this article

Osteosarcoma (OS) is the most frequent primary malignancy initially in bone with multiple genomic aberrations. Methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) is linked with the progression of diverse tumors. However, its function in OS is not understood completely. The expression pattern and prognostic significance of MTHFD1L in OS tissues were analyzed based on GEO database. The expression level of MTHFD1L in OS cell lines was explored by qRT-PCR. The cell proliferation, colony formation ability, invasion as well as migration in OS cells after MTHFD1L knockdown were determined using cell counting kit 8 (CCK-8) assay, colony formation and transwell methods. GSEA analysis was performed to predict the underlying mechanisms of MTHFD1L in OS development. Furthermore, the western blot was utilized to study the influence of MTHFD1L on AKT/mTOR pathway. Our results indicated that MTHFD1L expression was significantly up-regulated in OS tissues and cells compared with normal tissues and cells. High expression of MTHFD1L could lead to poor prognosis of OS patients. Cell proliferation, colony formation ability, migration and invasion were blocked because of reduced MTHFD1L

Objective: To analyze the levels of in vivo immunological indicators in patients with different types of malignanat lymphoma. Methods: A total of 69 patients with different types of malignanat lymphoma who were admitted in our hospital were included in the study, among which 31 had primary extranodular malignant lymphoma (extranodular group), and 38 had primary nodular malignant lymphoma (nodular group). Moreover, 34 patients with pure lymphnoditis were served as the control group. FCM and full-automatic speed scattering turbidimetry were used to detect T lymphocyte subsets proportion, IgG, IgA, IgM, C3, and C4 in the three groups. Results: The comparison of CD3+, CD4+, CD8+, NK cell proportion, and CD4+/CD8+ among the three groups was significantly different. When compared with the control group, CD3+, CD4+, CD8+, NK cell proportion, and CD4+/CD8+ in the extranodular group and nodular group were significantly reduced; moreover, CD3+, CD4+, CD8+, and CD4+/ CD8+ in the extranodular group were significantly lower than those in the nodular group, while NK cell in the extranodular group was significantly higher than that in the nodular group. The comparison of IgG, IgA, and C3 levels among the three groups was statistically significant. When compared with the control group, IgG and IgA levels in the extranodular group and nodular group were significantly elevated, while C3 level was significantly reduced, and the comparison between the extranodular group and nodular group between the two groups was statistically significant. IgM level in the extranodular group and nodular group was elevated, C4 level was reduced, but the comparison with the control group was not statistically significant. Spearman correlation analysis showed that CD3+ and CD4+ were significantly correlated with IgA, IgG, and C3. Conclusions: With the increasing of severity degree of lymphoma, the immunological level is reduced, while CD3+ and CD4+ are significantly correlated with IgA, IgG, and C3, indicating that detection of CD3+ and CD4+ proportion or IgA, IgG, and C3 contents in combined with pathological characteristics can evaluate the severity degree of malignant lymphoma in order to provide guidance for the clinical diagnosis and treatment.

Konjac glucomannan (KGM) electrogels were successfully prepared under alternating current (AC) in the presence of potassium chloride (KCl). The structure of the gels was studied using Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and scanning electron microscopy (SEM). A single-factor experiment was performed to optimize the preparation of the gels. Our results showed that KGM was degraded under AC and partially deacetylated. KGM and KCl formed the structure , and electrogels with porous structures retained some acetyl groups. Furthermore, as the KCl concentrations, voltages, time, and KGM concentrations increased, the viscoelastic moduli of the gels increased; the moduli decreased when the KCl concentrations, voltages, and time exceeded critical values.

This chapter introduces the basic principle of biomineralization and the theory of biological mineralization interface control, and the progress of natural bone formation and mineralization. Natural bone is composed of mineralized collagen (MC) by a series of self-assembly and molecular rearrangement and subsequent processing and modification of the formation. The natural bone formation and biological mineralization are closely related. Preliminary mineralization of collagen and its surface nucleation sites. Biomimetic preparation of hydroxyapatite MC fibers are a long-sought bone graft material in many laboratories because of their composition, structure, and properties. Natural bones are similar and have good bioactivity and biodegradability. There are many reports on the structure, properties, synthesis mechanism and application of collagen/HA and with the increasing acceptance of biomimetic materials, several collagen and hydroxyapatite composite materials have gained market access permits, and more and more doctors are accepting them.

Extensive evidence indicates that oxidative stress plays a pivotal role in the development of osteoporosis. We show that naringin, a natural antioxidant and anti-inflammatory compound, effectively protects human adipose-derived mesenchymal stem cells (hADMSCs) against hydrogen peroxide (H2O2)-induced inhibition of osteogenic differentiation. Naringin increased viability of hAMDSCs and attenuated H2O2-induced cytotoxicity. Naringin also reversed H2O2-induced oxidative stress. Oxidative stress induced by H2O2 inhibits osteogenic differentiation by decreasing alkaline phosphatase (ALP) activity, calcium content and mRNA expression levels of osteogenesis marker genes RUNX2 and OSX in hADMSCs. However, addition of naringin leads to a significant recovery, suggesting the protective effects of naringin against H2O2-induced inhibition of osteogenic differentiation. Furthermore, the H2O2-induced decrease of protein expressions of β-catenin and clyclin D1, two important transcriptional regulators of Wnt-signaling, was successfully rescued by naringin treatment. Also, in the presence of Wnt inhibitor DKK-1, naringin is no longer effective in stimulating ALP activity, increasing calcium content and mRNA expression levels of RUNX2 and OSX in H2O2-exposed hADMSCs. These data clearly demonstrates that naringin protects hADMSCs against oxidative stress-induced inhibition of osteogenic differentiation, which may involve Wnt signaling pathway. Our work suggests that naringin may be a useful addition to the treatment armamentarium for osteoporosis and activation of Wnt signaling may represent attractive therapeutic strategy for the treatment of degenerative disease of bone tissue.

The objective of this study was to investigate the prevalence of the cfr gene in Escherichia coli isolates from domestic animals in Northeast China and to characterize the cfr-containing plasmids. Between June 2015 and April 2016, 370 E. coli isolates were collected from pigs, chickens, and dairy cows in Northeast China. Among these, 111 were florfenicol resistant, including 109 isolates carrying the floR gene and 6 positives for cfr. The prevalence of cfr in E. coli isolates from the four northeast provinces in China was 1.6% (6/370), which was higher than that previously reported (0.08% and 0.5%). All six cfr-containing E. coli isolates were highly resistant to florfenicol (100%), cefotaxime (100%), and fosfomycin (100%). Complete sequence analysis of two cfr-carrying plasmids revealed high homology of the IncX4-type pEC14cfr plasmid with two other cfr-harboring plasmids, pSD11 and pGXEC6, found in swine E. coli isolates from southern China. pEC14cfr-like plasmids have been isolated in five provinces in southern and northern China. The isolation sites were up to 2700 kilometers apart, implying that pEC14cfr-like plasmids are likely to be national epidemic cfr-carrying plasmids that mediate the dissemination of cfr in China. Moreover, the genetic structure (IS26-IS26-cfr-rec-pre/mob-ramA-IS26) of the second cfr-carrying plasmid, IncF14:A-:B- pEC295cfr, represents a novel genetic environment for cfr identified for the first time in the present study. Sequence homology analysis indicated that the cfr-carrying element was most likely introduced into a cfr-negative pEC12 plasmid backbone, which evolved into the cfr-carrying vector, pEC295cfr. Moreover, isolation of the IncF14:A-:B- pEC295cfr plasmid harboring cfr suggests that IncFII plasmids maybe have become additional effective vehicles for cfr dissemination. These results highlight the importance of surveying the prevalence of IncX4 and IncFII plasmids in gram-negative bacteria, especially in swine E. coli isolates.

Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) is a facultative intracellular pathogen that causes huge losses in poultry industry and also food poisoning in humans due to its being a food-borne pathogen. Functions of Invasion-related genes need to be explored, as invasion is a key step for Salmonella infection. In this study, a transposon mutant library of Salmonella Enteritidis isolate SM6 was constructed and screened for the invasion-related genes via incubation with Caco-2 cells. Three stably attenuated mutants were identified for significantly reduced invasion with insertions all in hilA (hyperinvasive locus A) gene. We constructed and evaluated the hilA deletion mutant in vivo and in vitro. SM6△hilA showed significantly reduced ability to invade Caco-2 cells and decreased pathogenicity in chicks. However, the bacterial load and pathological damage in the cecum were significantly higher than those in the SM6 in vivo. Present results provide new evidences for pathogenicity research on Salmonella Enteritidis.

To construct a novel live, attenuated Salmonella vaccine, the lon, cpxR and cpdB genes were deleted from a wild-type Salmonella enterica serovar Enteritidis-6 (SM-6) strain using the phage λ Red homologous recombination system, resulting in SM-△CpxR, SM-△C/Lon and SM-△C/L/CpdB. The growth curves of strain SM-△C/Lon grew more rapidly than the other strains and had OD 600 values higher than the other strains starting at the 4 h time point. The growth curves of strain SM-△C/L/CpdB were relatively flat. The colonization time of SM-△C/L/CpdB is about 8-10 days. Deleting the lon/cpxR/cpdB (SM-6) genes resulted in an approximate 10(3)-fold attenuation in virulence assessed by the analysis of the LD50 of specific pathogen-free (SPF) chicks. This result indicated that the deletion of the lon, cpxR and cpdB genes induced significant virulence attenuation. The protective effects of SM-△C/L/CpdB vaccination in SPF chicks against 5 × 10(9) colony forming units (CFU) of S. Enteritidis were resulted from the induction of an effective immune response. These findings demonstrate the potential of mutant SM-△C/L/CpdB to be used as an effective vaccine.

Salmonella enterica serovar Pullorum (S. Pullorum) is a worldwide poultry pathogen of considerable economic importance, particularly in those countries with a developing poultry industry. A variety of genes that affect S. Pullorum colonization in chickens had been identified. 2′,3'-cyclic phosphodiesterase (cpdB) is the bifunctional enzyme which possess 2′,3'-cyclic phosphodiesterase as well as 3'-nucleotidase activity. To assess the role of cpdB of S. Pullorum in colonization of cecum and internal organs in poultry, seven-day-old chicks were infected with 109 CFU/ml of a cpdB mutant and wild type strain. High number of cpdB mutant and wild type strain colonized the internal organs shortly after infection, but no colonization of cpdB mutant were observed from internal organs at day 10 post-infection, meanwhile, wild type bacteria in internal organs were observed at day 16 post-infection. Furthermore, the colonization of cpdB mutant in the cecum was seriously decreased from 6 days post-infection simultaneously wild type strain was increased and seriously decreased at day 8 post-infection. At day 12 post-infection, no cpdB mutant was observed from cecum, however high numbers of wild type strain were isolated at day 16 post-infection. It is concluded that cpdB is involved in long-term colonization of S. Pullorum in the chicks' cecum and internal organs. In addition, deletion of cpdB from S. Pullorum was not affect the morphology and growth of bacteria.

At the time of implanting bone-related implants into human body, a variety of biological responses to the material surface occur with respect to surface chemistry and physical state. The commonly used biomaterials (e.g. titanium and its alloy, Co-Cr alloy, stainless steel, polyetheretherketone, ultra-high molecular weight polyethylene and various calcium phosphates) have many drawbacks such as lack of biocompatibility and improper mechanical properties. As surface modification is very promising technology to overcome such problems, a variety of surface modification techniques have been being investigated. This review paper covers recent advances in surface modification techniques of bone-related materials including physicochemical coating, radiation grafting, plasma surface engineering, ion beam processing and surface patterning techniques. The contents are organized with different types of techniques to applicable materials, and typical examples are also described.

To compare the therapeutic effects of acupuncture combined with fire dragon moxibustion and simple acupuncture therapy in the treatment of patients with cervical spondylotic radiculopathy (CSR) of kidney-deficiency-cold type.Ninety kidney-deficiency-cold type CSR outpatients were randomly divided into control (acupuncture,① After the treatment, CSR-20-points scores in both treatment and control groups were significantly increased in comparison with pre-treatment in each group (Acupuncture combined with fire dragon moxibustion is superior to simple acupuncture therapy in improving clinical symptoms of patients with CSR of kidney-deficiency-cold type, being a recommendable scheme in clinical practice.

Thirty-nine fosfomycin-resistant Escherichia coli isolates carrying fosA3 were obtained from pigs, chickens, dairy cows, and staff in four northeastern provinces of China between June 2015 and April 2016. The fosA3 gene was colocated with bla CTX-M genes on conjugative plasmids of the incompatibility groups IncN ( n = 12), IncN-F33:A − :B − ( n = 2), IncF33:A − :B − ( n = 14), IncF14:A − :B − ( n = 2), and IncI1/sequence type 136 (ST136) ( n = 9). Four different genetic contexts of fosA3 were detected among the 39 E. coli isolates. Three potential epidemic plasmids circulated among E. coli strains from this region.

Thirty-nine fosfomycin-resistant Escherichia coli isolates carrying fosA3 were obtained from pigs, chickens, dairy cows, and staff in four northeastern provinces of China between June 2015 and April 2016. The fosA3 gene was colocated with blaCTX-M genes on conjugative plasmids of the incompatibility groups IncN (n = 12), IncN-F33:A−:B−(n = 2), IncF33:A−:B−(n = 14), IncF14:A−:B−(n = 2), and IncI1/sequence type 136 (ST136) (n = 9). Four different genetic contexts of fosA3 were detected among the 39 E. coli isolates. Three potential epidemic plasmids circulated among E. coli strains from this region.

Objectives: To determine the complete nucleotide sequence of the multidrug resistance plasmid pSCEC2, isolated from a porcine Escherichia coli strain, and to analyse it with particular reference to the cfr gene region. Methods: Plasmid pSCEC2 was purified from its E. coli J53 transconjugant and then sequenced using the 454 GSFLX System. After draft assembly, predicted gaps were closed by PCR with subsequent sequencing of the amplicons. Results: Plasmid pSCEC2 is 135615 bp in size and contains 200 open reading frames for proteins of ≥100 amino acids. Analysis of the sequence of pSCEC2 revealed two resistance gene segments. The 4.4 kb cfr-containing segment is flanked by two IS256 elements in the same orientation, which are believed to be involved in the dissemination of the rRNA methylase genecfr. The other segment harbours the resistance genesfloR,tet(A)-tetR,strA/strB andsul2, which have previously been found on other IncA/C plasmids. Except for these two resistance gene regions, the pSCEC2 backbone displayed .99% nucleotide sequence identity to that of other IncA/C family plasmids isolated in France, Chile and the USA. Conclusions: The cfr gene was identified on an IncA/C plasmid, which is well known for its broad host range and transfer and maintenance properties. The location on such a plasmid will further accelerate the dissemination of cfr and co-located resistance genes among different Gram-negative bacteria. The genetic context of cfr on plasmid pSCEC2 underlines the complexity of cfr transfer events and confirms the role that insertion sequences play in the spread of cfr.

The collagenase cathepsin K has been shown important in the pathogenesis of rheumatoid arthritis (RA). Icariin is the major pharmacologically active flavonol diglycoside of Herba Epimedii, an herb used in Chinese traditional medicine to treat arthritis. We investigated whether icariin can inhibit the protease activity of cathepsin K and its effects on a murine model of collagen-induced arthritis (CIA). Six-week old female BALB/C mice were immunized with type II collagen and treated with vehicle alone icariin (25mg/kg) for 21 days; a control remained untreated. Serum concentrations of type I collagen C-terminal telopeptide (CTX-I) and cartilage oligomeric matrix protein (COMP) and urinary concentrations of deoxypyridinoline (DPD) were measured, and disease severity was assessed. Compared with immunized, untreated mice, immunized icariin-treated mice had significantly lower urinary DPD (~25%, p

The occurrence and counts of Listeria monocytogenes were investigated in a total of 526 retail raw food samples. All L. monocytogenes isolates were further analyzed by serotyping and antimicrobial susceptibility assays. The molecular basis of tetracycline resistance of each isolate and the genetic relatedness were determined. L. monocytogenes isolates were found in 12.4% (65/526) of the samples, with counts below 10 2 CFU/g. L. monocytogenes was most commonly isolated from pork (20%, 20/100), seafood (13.8%, 15/109), chicken (13.2%, 14/106), and beef (10.3%, 11/107). In addition, L. monocytogenes was also detected in 4.8% (5/104) of raw mutton samples. Four serogroups were identified among the 65 L. monocytogenes isolates, with serogroups 1/2a-3a (60%) and 4b-4d-4e (24.6%) being dominant. Most L. monocytogenes isolates were resistant to cefotaxime (54.6%), fosfomycin (51.5%), and clarithromycin (36.4%). Some isolates showed intermediate resistance to streptomycin (12.1%), norfloxacin (13.6%), ciprofloxacin (13.6%), and nitrofurantoin (9.1%). Multiple resistances were observed in 72.3% of isolates. Genetic relatedness analysis revealed that there were no prominent associations between specific food types, serotypes, antimicrobial susceptibility profiles and Pulsed-field gel electrophoresis (PFGE) patterns. In addition, these isolates were multiresistant and belonged to the epidemiologically important serotypes 1/2a and 4b, implying a potential public health risk.

In this study, a hybridoma-based technique and phage display technology were used to obtain mouse monoclonal antibodies (mAb) against cold-shock DEAD-box protein A (CsdA) from Mycobacterium tuberculosis and to determine the location of the relevant epitope. One highly specific mAb, named A3G5, was developed against the recombinant CsdA protein (rCsdA) and could detect rCsdA protein in enzyme-linked immunosorbent assays (ELISA) and Western blot assays. By screening a phage displayed library of random 12-mers (Ph.D.-12), 10 positive phage clones were randomly selected after three rounds of bio-panning and identified by ELISA. Eight of these clones were sequenced, and their amino acid sequences were deduced. One B-cell epitope (-APDPPLSRR-) in the rCsdA protein was identified with mAb A3G5. A synthetic peptide (-MAPDPPLSRR-) (Cpep) matched well with the CsdA sequence at 443–451 aa and was confirmed by affinity ELISA, competitive inhibition assays and the development of an immune response in mice. These results may be of great potential value in the further analysis of the function and structure of the CsdA protein from M. tuberculosis .

Assessing the inter-annual variation of regional grassland productivity is imperative to meet the local requirements of grassland adaptive management at regional- or landscape- scale. For the semiarid grassland of Inner Mongolia, the improved CASA model, a kind of light-energy-efficiency model, was used to simulate the Net Primary Productivity (NPP) of the regional grassland. And this study further calculated the Standard Deviation (SD) and Coefficient of Variation (CV) of the regional NPP. Both of SD and CV were used to reflect the fluctuations of regional NPP in the study area among years. Approximately 1/3 of the regional NPP over the years were dramatically changed, frequently up to large amplitude by an average rate of 1 times or more.

More and more applications of nanomaterials have been achieved in the biomedicine field. Numerous nanomedical devices, such as bone grafts with nano-hydroxyapatite and the silver-based anti-bacteria products, have been developed and have been trying to enter into the Chinese market. The State Food and Drug Administration of China (SFDA) is facing a critical challenge of how to explore and supervise the safety assessment of the nanomedical products. This paper briefly introduces the approval status of nanomedical products and the current advances of the safety assessment of nanomaterials in China.

It is a knotty problem in the actual project, that contraction deformation produce stretching stress on the concrete engineering. To counter deformation characteristic of the large-scale concrete foundation mat in construction temperature field,this paper has put a complete set of compute mechanics model of stress analysis and influence limits of constrained deformation on the basis of Saint-Venant’s principle. A number of computing formulas were inferred with theory of materil mechanics and structural mechanics,so that have obtained a set of analysis methods.By theory analysing and computing an actual project, it was that compute result accorded with structure law and the seat of compute crack was identical with cracking seat of the actual project.So, the achievement has the same values about the theory and the engineering application.

A simple, rapid, and selective method to determine the concentration of mildronate in human plasma and urine using ultra performance liquid chromatography–tandem mass spectrometry (UPLC-MS-MS) was developed and validated. The detection was performed on a triple-quadrupole tandem mass spectrometer by multiple reaction monitoring mode via electrospray ionization at m/z 147.2–58.0 for mildronate and m/z 147.2–87.8 for the internal standard, carbachol. The UPLC separation was carried out with a UPLC BEH HILIC column. The mobile phase consisted of 0.08% formic acid in 30 mM ammonium acetate solution and acetonitrile (23:77, v/v). Plasma samples were extracted from plasma by protein precipitation and urine samples were diluted with the mobile phase. The analysis time was 3.5 min for each sample. Linear calibration curves ranged from 0.10 to 100.00 μg mL−1 in human plasma and 0.50 to 600.00 μg mL−1 in urine. The method had been successfully applied to a pharmacokinetic study in healthy volunteers. After single intravenously administration of 250, 500, and 750 mg mildronate, the elimination half-life (t 1/2) were (2.74 ± 0.67), (4.86 ± 0.82) and (5.16 ± 0.77) h, respectively. The t 1/2 for the 250 mg dose did vary significantly with other dosages (P

A solid-phase extraction–liquid chromatographic–tandem mass spectrometry method for the determination of nalbuphine concentrations in human plasma has been developed. Samples (1 mL) were extracted using a Strata™-X solid phase extraction cartridges. Chromatographic separation of nalbuphine and naloxone (internal standard) was achieved on a Phenomenex Kinetex PFP (2.6 μm, 100 A, 100 × 2.1 mm) column using a mobile phase consisting of 0.1% formic acid, 15 mM ammonium acetate in deionized water and acetonitrile (60:40, v/v). The flow rate was 0.3 mL/min and the total run time was 2 min. Detection of the analytes was achieved using positive ion electrospray ionization via multiple reactions monitoring mode. The mass transitions were m/z 358 340 for nalbuphine and m/z 328 310 for naloxone. The assay was linear over the concentration range 0.50–500.00 ng/mL, with correlation coefficients ≥0.995. The lower limit of quantitation was set at 0.5 ng/mL plasma based on an average signal-to-noise ratio of 44.79. The intra- and inter-day precision was less than 8.07% in terms of relative standard deviation and accuracy ranged from 94.97 to 106.29% at all quality control levels. The method was applied successfully to determine nalbuphine concentrations in human plasma samples obtained from subjects receiving intravenous administration of nalbuphine. The method is rapid, sensitive, selective and directly applicable to human pharmacokinetic studies involving nalbuphine. Copyright © 2011 John Wiley & Sons, Ltd.

The nonlinear FEM models of concrete-filled square tubular column with internal diaphragms and steel beams with reduced beam section bolt-weld joints, involving geometric, materials and contact nonlinear, were established. The 3D models of beam-to-column joints with internal diaphragms by 3D solid elements are founded. The bearing capacity behavior of joints under the monotonic loading are analyzed, including P-Δ curves, the stresses distribution of beam, the dissipating energy ability and destruction form. The analysis results show that the initial stiffness of joint with reduced beam section is close to that of non-reduced beam section joint. The bearing capacities of joints with reduced beam section are lower a little than that of non-reduced beam section joint. The plastic hinge in the joint with reduced beam section is removed to the reduced beam region.

Avian pathogenic Escherichia coli (APEC) is an important respiratory pathogen of poultry. A variety of virulence-associated genes and serogroups are associated with avian colibacillosis caused by APEC strains. One hundred forty-eight E. coli isolates recovered from diagnosed cases of avian colibacillosis from Guangdong province between 2005 and 2008 were serotyped, and characterized for virulence-associated genes, phylogenetic backgrounds, antibiotic susceptibility, and genetic relatedness. Associations between virulence-associated genes and antimicrobial resistance were further analyzed. Although 148 APEC isolates belonged to 21 different serogroups, 81% were of one of eight serogroups: O65 (27%), O78 (10%), O8 (9%), O120 (9%), O2 (7%), O92 (6%), O108 (5%), and O26 (5%). Polymerase chain reaction analysis showed that the most prevalent gene was traT (90%), followed by iroN (63%), fimH (58%), hlyF (55%), cvaC (54%), and sitA (51%). The APEC strains mainly belonged to groups A (73%) and D (14%). Multiple antimicrobial-resistant phenotypes (greater than or equal to three antimicrobials) were detected in all E. coli isolates, with the majority of isolates displaying resistance to tetracycline (97%), sulfamethoxazole (93%) and fluoroquinolones (87% for ciprofloxacin and 84% for enrofloxacin), chloramphenicol (74%), and florfenicol (66%). All E. coli isolates were further genetically characterized by pulsed-field gel electrophoresis. A total of 125 different pulsed-field gel electrophoresis profiles were obtained, implying that the multiresistant E. coli isolates carrying virulence-associated genes and belonging to multiple serogroups were not derived from a specific clone, but represented a wide variety of chromosomal backgrounds. Statistical analysis showed that several virulence-associated genes were significantly present in APEC isolates susceptibility to multiple antimicrobials. The findings demonstrate that a wide variety of serogroups and potential virulence genes, multiple-resistances, and the clear association of susceptibility and virulence genes have commonly emerged in APEC strains, and these also suggest that antimicrobials should be prudently used to reduce the emergence and spread of resistant strains carrying virulence-associated genes.

Escherichia coli isolates from diseased pigs were examined for antimicrobial susceptibility to 12 antimicrobials and possession of virulence genes (VGs), and then grouped according to the phylogenetic background and genetic relatedness. Associations between antimicrobial resistance (AMR) and VGs and between AMR and phylogenetic group were subsequently assessed. The results showed that most isolates (91%) were epidemiologically unrelated. Multiple antimicrobial-resistant phenotypes (or=5 antimicrobials) were observed in 89% of E. coli strains and the most frequent types of resistance were to sulfamethoxazole (95%), tetracycline (94%), chloramphenicol (89%), and streptomycin (84%). The majority of isolates belonged to phylogenetic group A (84%). The most prevalent VG was EAST1 (64%), followed by Stx2e (63%) and eae (47%). Resistance to ceftiofur was associated with the presence of certain VGs, whereas resistance to doxycycline and kanamycin was associated with the absence of certain VGs. These findings suggest that multidrug resistance phenotypes, a variety of VGs, and the clear associations between resistance and VGs are commonly present in E. coli strains from diseased pigs. These results indicate that there is a great need for surveillance programs in China to monitor AMR in pathogenic E. coli strains.

According to physiological and biochemical characteristics of Leptospirillum ferriphilum, a strain of object bacteria was isolated successfully. Bacteria were enriched by selective liquid medium and plated on designed single-layered agar solid medium. Colony was cultured and bacteria were collected. The morphologies of the object bacteria were observed using crystal violet staining, scanning electron microscope(SEM) and transmission electron microscope (TEM). The result of 16S rDNA identification shows that this bacterium belongs to Leptospirillum ferriphilum and it is named as Leptospirillum ferriphilum strain D1. These results indicate that this new single-layered agar solid medium is efficient and simple for isolation of Leptospirillum ferriphilum. Additionally, physiological-biochemical characteristics show that the optimum initial pH value and its growth temperature are 1.68 and 40 °C, respectively. The culture of it is used to leach a complex concentrate chalocpyrite, the leaching efficiencies of copper and iron are 1.93% and 13.74%, respectively, and it is more effective than the A. ferrooxidans culture in the leaching of the complex concentrate chalcopyrite.

A novel nonconjugative plasmid of 28,489 bp from a porcine linezolid-resistant Enterococcus faecium isolate was completely sequenced. This plasmid harbored a novel type of multiresistance gene cluster that comprised the resistance genes lnu (B), lsa (E), spw , aadE , aphA3 , and two copies of erm (B), which account for resistance to macrolides, lincosamides, streptogramins, pleuromutilins, streptomycin, spectinomycin, and kanamycin/neomycin. Structural comparisons suggested that this plasmid might have developed from other enterococcal plasmids by insertion element (IS)-mediated interplasmid recombination processes.

Two porcine Escherichia coli isolates harbored the cfr gene on conjugative plasmids of 38,405 bp (pGXEC6) and 41,646 bp (pGXEC3). In these two plasmids, the cfr gene was located within a 4,612-bp region containing a tnpA -IS 26-cfr -IS 26 -Δ hyp element. Plasmid pGXEC3 was almost identical to pGXEC6 except for a 3,235-bp IS Ecp1-bla CTX-M-14b insertion. The colocation of the multiresistance cfr gene with an extended-spectrum-β-lactamase gene on a conjugative plasmid may support the dissemination of these genes by coselection.

The dynamic expression pattern analysis provides the primary information of gene function. Differences of the RNA and/or protein location will provide valuable information for gene expression regulation. Generally, in situ hybridization (ISH) and immunohistochemistry (IHC) are two main techniques to visualize the locations of gene transcripts and protein products in situ, respectively. Here we describe the protocol for the whole brain dissection, the in situ hybridization, and the immunostaining of the developing Xenopus brain sections. Additionally, we point out the modification of in situ hybridization for microRNA expression detection.

This study evaluated maternal immunity against Salmonella enterica serovar Enteritidis acquired through the egg yolk. Two-hundred 19-week-old specific pathogen free (SPF) broiler breeders which were randomly divided into two groups of equal size were injected with S. Enteritidis ghosts (5 × 10(9) colony forming units in 0.1 ml per hen) and phosphate-buffered saline (PBS, 0.01 mol ⋅ l(-1), pH 7.4) twice, respectively, with an interval of 2 weeks. An indirect enzyme-linked immunosorbent assay (ELISA) was applied to detect specific antibodies against S. Enteritidis. S. Enteritidis-specific antibody levels in the vaccinated group increased over time and were significantly higher than those of the control group on days 28 (P0.001) and 35 (P0.001) post-vaccination. Ten 7-day-old chicks from hens that were vaccinated with a S. Enteritidis ghost vaccine were challenged at 14 days of age with 5 × 10(9) CFU of S. Enteritidis DH091 (homologous to the vaccine strain), 8/10 (80%) chicks from vaccinated hens survived, whereas 3/10 (30%) chicks from unvaccinated hens survived. The chicks acquired high levels of serum antibodies against S. Enteritidis. These results reveal that maternal antibodies in chicks acquired from vaccinated hens through eggs can confer a significant protection against S. Enteritidis infection.

Increased knowledge of molecular alterations involved in gastric carcinogenesis has provided useful information for diagnosis and prediction. In this study, we investigated the clinicopathological significance of TFF3 expression and Her-2/neu status in gastric adenocarcinoma and explored the correlation between TFF3 expression and Her-2/neu status. A hundred and twenty-six (126) patients having undergone curative gastrectomy were enrolled. Immunohistochemistry for TFF3 was performed on tumor tissues and non-neoplastic resection margins. Her-2/neu status was assessed by immunohistochemical staining and fluorescence in situ hybridization (FISH) on tumor tissues. As a result, TFF3 expression and Her-2/neu positivity were detected in 46.8% and 11.9% of the cases, respectively. Patients with negative TFF3 exhibited longer survival than the positive group (P=0.0142), while patients with positive Her-2/neu only showed a tendency toward longer overall survival (P>0.05). However, Her-2/neu was significantly associated with improved disease-free survival (P=0.0234). Multivariate analysis demonstrated Her-2/neu and TFF3 to be independent prognostic indicators of recurrence, and a significantly poor prognosis for expression of TFF3 in patients with Her-2/neu negative tumors. TFF3 is an independent indicator for overall survival in gastric cancer, while Her-2/neu, as a marker of long time survival prediction, seems to be limited.

The aim of this study was to investigate the expression of nucleolin in tumorous tissues and corresponding non-malignant tissues in gastric cancer (GC), and the correlation of different expression sites with clinicopathologic parameters and prognosis. Immunohistochemistry was used for detecting the expression levels of nucleolin in GC tissues and corresponding non-malignant tissues from 124 gastrectomy specimens with stage I-III. Staining results were correlated with clinicopathologic features and survival. Both GC tissues and corresponding non-malignant tissues showed nucleolar staining for nucleolin. Nucleolin expression was higher in GC tissues than in non-malignant tissues. Among the 124 GCs, 85 (68.5%) were nucleolin-high. No significant correlation between nucleolin expression and other clinicopathologic parameters was found. The Cox univariate analysis indicated that both cytoplasmic staining and nucleolar staining of nucleolin expression correlated with patients' prognosis (log-rank, p < 0.0001; p = 0.0075, respectively). It was concluded in the study that nucleolin was overexpressed in GCs. A high level of nucleolar expression of nucleolin was an independent prognostic marker for better survival while high cytoplasmic staining was closely associated with worse prognosis for GC patients.