Refracture after plate removal of midshaft clavicle fractures after bone union—incidence, risk factors, management and outcomes

Abstract Introduction There is a great debate on the routine use of open reduction and internal fixation (ORIF) for midshaft clavicle fractures, and one concern is the adverse events after ORIF, such as implant removal after bone union. In this retrospective study, we assessed the incidence, risk factors, management and outcomes of refracture after plate removal of midshaft clavicle fractures after bone union. Materials and methods Three hundred fifty-two patients diagnosed with acute midshaft clavicle fractures who had complete medical records from primary fractures to refracture were recruited. Details of imaging materials and clinical characteristics were carefully reviewed and analysed. Results The incidence rate of refracture was 6.5% (23/352), and the average interval from implant removal to refracture was 25.6 days. Multivariate analysis showed that the risk factors were Robinson type-2B2 and fair/poor reduction. Females were 2.4 times more likely to have refracture, although it was not significant in multivariate analysis (p = 0.134). Postmenopausal females with a short interval (≤ 12 months) from primary surgery to implant removal had a significant risk for refracture. Tobacco use and alcohol use during bone healing were potential risk factors for male patients, although they were not significant in multivariate analysis. Ten patients received reoperation with or without bone graft, and they had a higher rate of bone union than 13 patients who refused reoperation. Conclusion The incidence of refracture following implant removal after bone union is underestimated, and severe comminute fractures and unsatisfactory reduction during primary surgery are risk factors. Implant removal for postmenopausal female patients is not recommended due to a high rate of refracture

Enhancing the secretion pathway maximizes the effects of mixed feeding strategy for glucose oxidase production in the methylotrophic yeast Pichia pastoris

Abstract Background A mixed feeding strategy (co-feeding of complex carbon sources with methanol) has become a common practice for process development in Pichia pastoris to increase cell biomass and enzyme production levels. However, in some cases mixed feeding did not have a significant impact or even had a negative effect on specific enzyme productivity. We hypothesized that this may be due to a bottleneck in the protein secretion pathway caused by too strong protein expression as a result of mixed feeding operation. Results Using glucose oxidase (Gox) as a model protein, the individual and synergistic effects of co-feeding of sorbitol or yeast extract (YE) with methanol and Hac1p overexpression on the secretory expression of Gox were investigated both in shake flasks and in a laboratory fermenter. The results showed that YE is superior to sorbitol in terms of stimulating protein expression and cell growth. Moreover, separate applications of the mixed feeding strategy and secretory pathway engineering only achieved limited success in enhancing Gox levels, while the combined use of the two strategies acted synergistically, leading to 297% increase of Gox production and the final enzyme titer reached 787.4 U/mL in GSgox-Pp on 1-L fermenter. Conclusions Co-feeding of YE combined with secretion pathway engineering significantly improved glucose oxidase secretion, which can be also applied to improve secretory expression of other foreign proteins in P. pastoris system

Additional file 1 of Refracture after plate removal of midshaft clavicle fractures after bone union—incidence, risk factors, management and outcomes

Supplementary Material

Clinical application, evaluation and analysis of influencing factors analysis of on tuberculosis ?-Interferon enzyme-linked immunosorbent assay

Objective: To assess sensitivity and specificity of the interferon gamma release assay test, and to pinpoint the influencing factors that should be taken care of in clinical application. Methods: The study was conducted at the First People's Hospital in the Yunnan Province of China from October 2018 to March 2019, and comprised samples collected from outpatient and inpatients. To detect mycobacterium tuberculosis, acid-fast staining on sputum smear was performed on relevant tissues suspected of extrapulmonary tuberculosis. Tuberculosis interferon gamma release assay test and pathology samples were examined. Tuberculosis-specific cell reaction assay kit was used for sampling. SysmexXN-2000 haematology analyser, VACUETTE SRS100/II and Beckman Coulter AU5800 were used to perform various analyses. Data was grouped and analysed using R statistical software. Results: Of the 960 samples, 516(53.75%) cases tested positive for tuberculosis infection and 444(46.25%) tested negative. The sensitivity of the pathological results was 86% and the specificity was 96%. The sensitivity of the interferon gamma release assay test was 95% and specificity 82%. Interferon release test, pathological results and final diagnosis showed significant comparisons (p<0.05). Significant relationships were also established for factors, such as age, interferon release quantity, lymphocyte, C-reactive protein and counts of mono-nuclear cell (p<0.05). Conclusions: Interferon gamma release assay test was found to have high consistency with pathological results and final diagnosis and can be used as a subsidiary to traditional clinical imaging and pathological judgment. Key Words: Tuberculosis, Interferon release, CRP, Hormone use. Continuous...

A new transient expression system for large-scale production of recombinant proteins in plants based on air-brushing an Agrobacterium suspension

Plant transient expression using virus-based vectors is advantageous when high level of gene expression is desired within a short time. In this study, a new system, named “air-brush,” has been developed to facilitate a scale-up production of recombinant proteins in plants. GFP was expressed successfully in Nicotiana benthamiana (Nb) plants by air-brushing an Agrobacterium suspension that contained the TMV-based vector p35S-30B-GFP. Key factors influencing the gene expression were optimized, including the Agrobacterium cell density, seedling age, and the growth temperature of plant materials. In addition, the pharmaceutical protein human acidic fibroblast growth factor (ha FGF) was also expressed in Nb plants by the air-brush system. The results demonstrated that using this system is highly advantageous; it is convenient, quick, easily scaled-up, and has a higher expression efficiency than leaf infiltration

MOESM1 of Enhancing the secretion pathway maximizes the effects of mixed feeding strategy for glucose oxidase production in the methylotrophic yeast Pichia pastoris

Additional file 1: Table S1. The oligonucleotides used in this study. Figure S1. Relative transcription levels of Gox with different co-feeding strategies. Control: GSgox cultured on methanol; YE: Gsgox cultured on mixed methanol and yeast extract (5:1 g/g); Sorbitol: Gsgox cultured on mixed methanol and sorbitol (5:1 g/g)

Enhancement of the Gene Targeting Efficiency of Non-Conventional Yeasts by Increasing Genetic Redundancy

National Natural Science Foundation of China [31000026]; Knowledge Innovation Program of the Chinese Academy of Sciences [KSCX2-EW-G-15-03]In contrast to model yeasts, gene targeting efficiencies of non-conventional yeasts are usually low, which greatly limits the research and applications of these organisms. In this study, we aimed to enhance the gene targeting efficiency of non-conventional yeasts by improving the fitness of mutant strains, particularly by increasing the genetic redundancy of host cells. To demonstrate this process, OCH1 gene deletion in Pichia pastoris was performed. Extra copies of the OCH1 gene on a helper plasmid were provided for the P. pastoris GS115 strain before the native OCH1 gene in the genomic DNA was knocked out. The redundancy in OCH1 gene significantly eliminated the growth defects of the och1 mutant and increased the deletion efficiency of the OCH1 gene by two orders of magnitude with the same length of homologous flanks. The same strategy was used to delete the KU70 and SGS1 genes. The targeting efficiencies of KU70 and SGS1 were increased by 1- and 23-fold, respectively. Therefore, this study provided an efficient strategy for the deletion of "stubborn" genes in non-conventional yeasts. This study further showed that cellular fitness is potentially an important factor that can limit the efficiency of gene targeting

Treatment-Related Prognostic Factors in Managing Osteosarcoma around the Knee with Limb Salvage Surgery: A Lesson from a Long-Term Follow-Up Study

Purpose. The aim of this study was to assess the treatment-related factors associated with local recurrence and overall survival of patients with osteosarcoma treated with limb-salvage surgery. Patients and Methods. Treatment-related factors were analyzed to evaluate their effects on local recurrence-free survival (LRFS) and overall survival (OS) in 182 patients from 2004 to 2013. Results. The mean length of follow-up was 73.4 ± 34.7 months (median, 68 months; range, 12-173 months), and 63 patients died by the end of the follow-up. The 5-year and 10-year overall survival rates were 68.6 ± 6.6% and 59.4 ± 10.6%, respectively. Univariate analysis showed that treatment-related prognostic factors for overall survival were prolonged symptom intervals >=60 days, biopsy/tumor resection performed by different centers, previous medical history, incomplete preoperative chemotherapy (21 days. In the multivariate analysis, biopsy/tumor resection performed by different centers, incomplete implementation of planned new adjuvant chemotherapy, and delayed resumption of postoperative chemotherapy (>21 days) were risk factors for poor prognosis; biopsy/tumor resection performed by different centers and tumor necrosis <90% were independent predictors of local recurrence. Conclusion. For localized osteosarcoma treated with limb-salvage surgery, it is necessary to optimize timely standard chemotherapy and to resume postoperative chemotherapy to improve survival rates. Biopsies should be performed at experienced institutions in cases of developing local recurrence

Schematic representation of the new strategy for efficient gene targeting.

<p>Take deletion of <i>OCH1</i> gene as an example. As follows, it’s a three-step process: 1) Generation of the transition host. The helper plasmid, which has the cloned <i>Pichia pastoris OCH1</i> gene placed under the control of the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter, was transformed into <i>P. pastoris</i> GS115 strain, thereby generating the transition host for <i>OCH1</i> gene deletion. The helper plasmid can be maintained within the host cell because of the presence of an autonomous replication sequence (PARS2). 2) Gene disruption. The disruption plasmid for <i>OCH1</i>, which contains the 5′ and 3′ flanking regions of the <i>OCH1</i> gene, was linearized and transformed into the transition host for gene deletion. 3) Elimination of the helper plasmid. After the chromosomal <i>OCH1</i> gene was successfully deleted, cells were streaked on an agar plate with methanol to induce the expression of <i>E. coli mazf</i> gene (encoding MazF). MazF toxicity causes a strong selection pressure on the streaked strains and induces the loss of the helper plasmids.</p