Search

Individual fish are vulnerable in hydropower reservoirs due to spillway and intake operations. It is essential to understand how reservoir forebay fish ecosystems respond to water levels, intake, and spillway regulation. This study aims to explore the fish entrainment risk of the Dawei Reservoir operation on the Dadu River in Sichuan, China, accounting for both intake and spillway operations under wet, normal, and dry seasonal reservoir water levels. Hydrodynamic variables, reservoir operation scenarios, and two fish species were used as indexes to analyze the fish entrainment risk. The simulation results showed that the fish entrainment risk was low under the Dawei intake operation schemes ranging from 0.84 × 103 to 5.97 × 103 m2. The results also showed that the fish entrainment risk was very high under the Dawei spillway operation in fish entrainment areas ranging from 3.90 × 104 to 2.08 × 105 m2. Based on the simulation results, the lowest fish entrainment risk happened with two intakes open and the reservoir water level at 2,640 m. The highest fish entrainment risk happened with five intakes open and the reservoir water level at 2,670 m. The results indicate that the long-term Dawei Reservoir regulation would not modify the fish entrainment risk at significant levels under the Dawei Reservoir operation schemes. HIGHLIGHTS A model was proposed to determine the fish entrainment risk.; The fish entrainment risk was low during the seasonal operation of intakes.; The maximum and minimum fish entrainment risks were established.; The fish entrainment risk was high under the spillway operation.; The long-term reservoir operation would not significantly affect the fish entrainment risk.;

Abstract Background Perioperative chemotherapy (ChT) and preoperative chemoradiation (CRT) are both the standard treatments for locally advanced gastric cancer (LAGC). CRT can achieve a higher pathological complete regression (pCR) rate, but whether this higher pCR rate can be transformed into a long-term survival benefit remains inconclusive. Therefore, relevant studies are in progress. On the other hand, immunotherapy has been established for the first-line treatment of advanced gastric cancer (AGC) and has been widely explored in the perioperative setting. The combination of chemotherapy/radiotherapy and immunotherapy may have a synergistic effect, which will lead to a better antitumor effect. The preliminary reports of ongoing studies show promising results, including a further improved pCR rate. However, the preferred treatment combination for LAGC is still not established. To solve this problem, we are carrying out this randomized phase II trial, which aims to evaluate the efficacy and safety of perioperative chemotherapy plus the use of PD-1 antibody with or without preoperative chemoradiation for LAGC. Methods Eligible patients with LAGC or gastroesophageal junction (GEJ) adenocarcinoma were randomized to receive perioperative ChT, PD-1 antibody, surgery with (Arm A) or without preoperative CRT (Arm B), and PD-1 antibody maintenance until one year after surgery. The primary endpoint of this study is that the pCR rate of Arm A will be significantly higher than that of Arm B. The secondary endpoints include the pathological partial regression (pPR) rate, R0 resection rate, objective response rate (ORR), event-free survival (EFS), overall survival (OS), safety and surgical complications. Moreover, several explorative endpoints will be evaluated to find and validate the predictive biomarkers of immunotherapy. Discussion The results of the NeoRacing study will provide important information concerning the application of PD-1 antibody in LAGC patients during the perioperative setting. Meanwhile, the two treatment protocols will be compared in terms of efficacy and safety. Trial registration ClinicalTrials.gov , NCT05161572 . Registered 17 December 2021 - Retrospectively registered.

ObjectivesPeriprosthetic joint infection (PJI) diagnosis remains challenging, and the identification of the causative microorganism is, by far, the most important aspect. Here, we use multiple PCR-based targeted next-generation sequencing (tNGS) to detect pathogens in PJI. To explore 1. the ability of targeted next-generation sequencing (tNGS) to detect pathogens in PJI; 2. the consistency of tNGS, metagenomic NGS (mNGS), and culture results; and 3. the ability of tNGS to detect drug resistance genes in PJI.MethodsPJI was diagnosed according to the Musculoskeletal Infection Society (MSIS) criteria. The microorganisms were detected by culture, mNGS and tNGS to compare the diagnostic effectiveness of the three methods for PJI and to compare their consistency in detecting microorganisms. Drug resistance genes were detected using tNGS. The costs and turnaround times of mNGS and tNGS were compared.ResultsForty-three patients with PJI, 21 patients without PJI and 10 negative control cases were included. The culture, tNGS, and mNGS sensitivities for PJI diagnosis were 74.41%, 88.37%, and 93.02%, respectively, with no significant differences. The specificities were 90.48%, 95.24%, and 95.24%, respectively, with no significant differences. tNGS detected drug resistance genes in 37.5% of culture-positive PJIs. tNGS was superior to mNGS for turnaround time (14.5 h vs. 28 h) and cost ($150 vs. $260).ConclusionstNGS can effectively identify PJI pathogens and may provide drug resistance information, while tNGS is superior to mNGS regarding cost and turnaround time. A multidisciplinary, multi-technology based algorithm to diagnose PJI is appropriate.Highlights298 microorganisms and 86 drug resistance genes were included in the tNGS panel.Diagnostic efficacy of tNGS is not inferior to that of commonly used indicators.tNGS is superior to mNGS in cost and turnaround time.

IntroductionThe diagnosis of Mycoplasma periprosthetic joint infection (PJI) is rather difficult due to its rarity and difficult in isolation, there are not standardized diagnostic procedure for Mycoplasma PJI presently. This study aimed to reported a metagenomic next-generation sequencing (mNGS)-based diagnostic strategy for Mycoplasma PJI.MethodsIn the present study, we have reported the largest number of Mycoplasma PJI that were precisely diagnosed by mNGS and verified by optimized microbial culture methods and (or) 16S PCR polymerase chain reaction (PCR).ResultsThe positive rate of optimized microbial culture methods and 16S PCR in the detection of Mycoplasma PJI was 57.14% and 71.43%, respectively. The infections were well controlled by targeted treatment in all cases.ConclusionThe standardized and optimized procedure based on mNGS presented in this study is useful for the diagnosis of Mycoplasma PJI, which might also be provided as a novel diagnostic strategy for rare bacterial PJI.

Abstract Synthetic biology has boosted the rapid development on using non-methylotrophy as chassis for value added chemicals production from one-carbon feedstocks, such as methanol and formic acid. The one-carbon dissimilation pathway can provide more NADH than monosaccharides including glucose, which is conducive for reductive chemicals production, such as succinic acid. In this study, the one-carbon dissimilation pathway was introduced in E. coli Suc260 to enhance the succinic acid production capability. Through the rational construction of methanol dissimilation pathway, the succinic acid yield was increased from 0.91 to 0.95 g/g with methanol and sodium formate as auxiliary substrates in anaerobic fed-batch fermentation. Furthermore, the metabolic flux of by-product pyruvate was redirected to succinic acid together with the CO2 fixation. Finally, through the immobilization on a specially designed glycosylated membrane, E. coli cells are more resistant to adverse environments, and the final yield of succinic acid was improved to 0.98 g/g. This study proved the feasibility of endowing producers with methanol dissimilation pathway to enhance the production of reductive metabolites. Graphical Abstract

The production of bio-based succinic acid through microbial CO2 fixation and conversion has gained significant attention as a promising approach to mitigate greenhouse gas emissions. However, the low CO2 utilization efficiency limits the efficient biosynthesis of succinic acid. Therefore, it is crucial from environmental and economic perspectives to enhance the efficiency of CO2 utilization in bio-succinic acid production. This review comprehensively covers the introduction of biosynthetic pathways for microbial CO2 fixation and the conversion of CO2 to succinic acid, as well as the challenges associated with CO2 supply and utilization effectiveness. Moreover, strategies including genetic and metabolic engineering for CO2 fixation, extracellular supply methods of CO2 and some potential technical approaches for CO2 capture (such as micro-nano bubbles, CO2 adsorption material and biofilm) are summarized and presented.

Objectives To characterize the abundance and relative composition of seawater bacterioplankton communities in Changle city using Illumina MiSeq sequencing and bacterial culture techniques. Methods Seawater samples and physicochemical factors were collected from the coastal zone of Changle city on 8 September 2019. Nineteen filter membranes were obtained after using a suction filtration system. We randomly selected eight samples for total seawater bacteria (SWDNA group) sequencing and three samples for active seawater bacteria (SWRNA group) sequencing by Illumina MiSeq. The remaining eight samples were used for bacterial culture and identification. Alpha diversity including species coverage (Coverage), species diversity (Shannon–Wiener and Simpson index), richness estimators (Chao1), and abundance‐based richness estimation (ACE) were calculated to assess biodiversity of seawater bacterioplankton. Beta diversity was used to evaluate the differences between samples. The species abundance differences were determined using the Wilcoxon rank‐sum test. Statistical analyses were performed in R environment. Results The Alpha diversity in the SWDNA group in each index was ACE 3206.99, Chao1 2615.12, Shannon 4.64, Simpson 0.05, and coverage 0.97; the corresponding index was ACE 1199.55, Chao1 934.75, Shannon 3.49, Simpson 0.09, and coverage 0.99. The sequencing results of seawater bacterial genes in the coastal waters of Changle city showed that the phyla of high‐abundance bacteria of both the SWDNA and SWRNA groups included Cyanobacteria, Proteobacteria, and Bacteroidetes. The main classes included Oxyphotobacteria, Alphaproteobacteria, and Gammaproteobacteria. The main genera included Synechococcus CC9902, Chloroplast, and Cyanobium_PCC‐6307. Beta diversity analysis showed a significant difference between the SWDNA and SWRNA groups (P

Gastrectomy can improve the clinical survival rate of gastric cancer patients, while the metabolic disorders caused by gastrectomy, seriously affect the postoperative recovery. In this study, probiotics (including Bifidobacterium infantis, Lactobacillus acidophilus, Enterococcus faecalis and Bacillus cereus) were used before gastrectomy to explore the effect of probiotics on gastrectomy patients. Our outcomes revealed probiotics could alleviate postoperative inflammation (Leukocyte; P

ObjectiveMany observational studies have found an association between Alzheimer’s disease (AD) and osteoporosis. However, it is unclear whether there is causal genetic between osteoporosis and AD.MethodsA two-sample Mendelian randomization (MR) study was used to investigate whether there is a causal relationship between osteoporosis and AD. Genes for osteoporosis and AD were obtained from published the genome-wide association studies (GWAS). Single nucleotide polymorphisms (SNPs) with significant genome-wide differences (p

BackgroundThe relationship between Ankylosing Spondylitis (AS) and the risk of stroke is complex. Therefore, we utilized Two-Sample Mendelian randomization to examine the probable causal link between these two features.MethodsThe genetic instruments linked to AS were chosen from a summary-level genetic data set from the FinnGen consortium in people of European ancestry (1462 cases and 164,682 controls). Stroke and its subtypes were selected as outcomes, and the MEGASTROKE consortium population was used to identify the genetic associations of AS on stroke (40,585 cases and 406,111 controls), ischemic stroke (IS) (34,217 cases and 406,111 controls), and its subtypes including large artery stroke (LAS) (4373 cases and 146,392 controls), small vessel stroke (SVS) (5386 cases and 192,662 controls), and cardioembolic stroke (CES) (7193 cases and 204,570 controls). Intracerebral hemorrhage (ICH) (1687 cases and 201,146 controls) data set from the FinnGen consortium was also used. To obtain the casual estimates, the inverse variant weighted (IVW) method was mainly used. By examining the heterogeneity and pleiotropy of particular single nucleotide polymorphisms (SNPs), the robustness of the results was also examined.ResultsThere was no evidence found to prove the correlation between genetically predicted AS and stroke (odds ratio [OR] 1.014; 95% confidence interval [CI] 0.999-1.031; P = 0.063), ICH (OR 1.030; 95% CI 0.995-1.067; P = 0.090), and IS (OR 1.013; 95% CI 0. 998-1.030; P = 0.090). In terms of the different subtypes of IS, there was strong evidence of positive causal inferences on CES (OR 1.051; 95% CI 1.022-1.081; P = 0.001), and suggestive evidence of positive causal inferences on LAS (OR 1.042; 95% CI 1.003-1.082; P = 0.033), while it was not significant for SVS (OR 1.010; 95% CI 0.975-1.047; P = 0.563).ConclusionThis study suggests that the possible causative impact of genetically predicted AS on stroke may be restricted to the CES and LAS subtypes.

Summary: Background: Early detection and prognosis prediction of colorectal cancer (CRC) can significantly reduce CRC-related mortality. Recently, circulating tumour DNA (ctDNA) methylation has shown good application foreground in the early detection and prognosis prediction of multiple tumours. Methods: This multicentre cohort study evaluated ctDNA methylation haplotype patterns based on archived plasma samples (collected between 2010 and 2018) from 1138 individuals at two medical centres: Fudan University Shanghai Cancer Center (Shanghai, China) and Southern Medical University Nanfang Hospital (Guangzhou, Guangdong, China), including 366 healthy individuals, 182 patients with advanced adenoma (AA), and 590 patients with CRC. Samples were processed using the ColonES assay, a targeted bisulfite sequencing method that detects ctDNA methylation haplotype patterns in 191 genomic regions. Among these 1138 samples, 748 were used to develop a classification model, and 390 served as a blinded cohort for independent validation. The study is registered at https://register.clinicaltrials.gov with the unique identifier NCT03737591. Results: The model obtained from unblinded samples discriminated patients with CRC or AA from normal controls with high accuracy. In the blinded validation set, the ColonES assay achieved sensitivity values of 79.0% (95% confidence interval (CI), 66%–88%) in AA patients and 86.6% (95% CI, 81%–91%) in CRC patients with a specificity of 88.1% (95% CI, 81%–93%) in healthy individuals. The model area under the curve (AUC) for the blinded validation set was 0.903 for AA samples and 0.937 for CRC samples. Additionally, the prognosis of patients with high preoperative ctDNA methylation levels was worse than that of patients with low ctDNA methylation levels (p = 0.001 for relapse-free survival and p = 0.004 for overall survival). Interpretation: We successfully developed and validated an accurate, noninvasive detection method based on ctDNA methylation haplotype patterns that may enable early detection and prognosis prediction for CRC. Funding: The Grant of National Natural Science Foundation of China (No.81871958), National Natural Science Foundation of China (No. 82203215), Shanghai Science and Technology Committee (No. 19140902100), Scientific Research Fund of Fudan University (No.IDF159052), Shanghai Municipal Health Commission (SHWJRS 2021-99), and Shanghai Sailing Program (22YF1408800).

Aims: To explore the effect of different durations of antibiotics after stage II reimplantation on the prognosis of two-stage revision for chronic periprosthetic joint infection (PJI). Methods: This study involved a retrospective collection of patients who underwent two-stage revision for chronic PJI and continued to use extended antibiotic prophylaxis in two regional medical centres from January 2010 to June 2018. The patients were divided into a short (≤ one month) or a long (> one month) course of treatment based on the duration of antibiotics following stage II reimplantation. The difference in the infection control rate between the two groups was compared, and prognostic factors for recurrence were analyzed. Results: A total of 105 patients with chronic PJI were enrolled: 64 patients in the short course group and 41 patients in the long course group. For 99 of the patients, the infection was under control during a follow-up period of at least 24 months after two-stage revision. For the short course group, the mean duration of antibiotic prophylaxis after stage II reimplantation was 20.17 days (SD 5.30) and the infection control rate was 95.3%; for the long course group these were 45.02 days (SD 15.03) and 92.7%, respectively. There was no significant difference in infection control rates between the two groups (p = 0.676). Cox regression analysis found that methicillin-resistant staphylococcus infection (p = 0.015) was an independent prognostic factor for recurrence. Conclusion: After stage II reimplantation surgery of two-stage revision for chronic PJI, extended antibiotic prophylaxis for less than one month can achieve good infection control rate. Cite this article: Bone Joint Res 2021;10(12):790–796.

Abstract Background Butyl acetate has shown wide applications in food, cosmetics, medicine, and biofuel sectors. These short-chain fatty acid esters can be produced by either chemical or biological synthetic process with corresponding alcohols and acids. Currently, biosynthesis of short chain fatty acid esters, such as butyl butyrate, through microbial fermentation systems has been achieved; however, few studies regarding biosynthesis of butyl acetate were reported. Results In this study, three proof-of-principle strategies for the one-pot butyl acetate production from glucose through microbial fermentation were designed and evaluated. (1) 7.3 g/L of butyl acetate was synthesized by butanol-producing Clostridium acetobutylicum NJ4 with the supplementation of exogenous acetic acid; (2) With the addition of butanol, 5.76 g/L of butyl acetate can be synthesized by acetate-producing Actinobacillus succinogenes130z (ΔpflA); (3) Microbial co-culture of C. acetobutylicum NJ4 and A. succinogenes130z (ΔpflA) can directly produce 2.2 g/L of butyl acetate from glucose by using microbial co-culture system with the elimination of precursors. Through the further immobilization of A. succinogenes130z (ΔpflA), butyl acetate production was improved to 2.86 g/L. Conclusion Different microbial mono- and co-culture systems for butyl acetate biosynthesis were successfully constructed. These strategies may be extended to the biosynthesis of a wide range of esters, especially to some longer chain ones.

ObjectivesMusculoskeletal pain (MSP) is a major reason for consultation in primary care and is becoming increasingly prevalent among medical students. There is little research on the current situation of MSP among Chinese medical and dental students. Data on the analysis of risk factors related to MSP are also limited. The objectives of this study were to investigate the prevalence and characteristics of MSP among Chinese medical and dental students and to explore the risk factors for MSP and students' intent to seek medical treatment.MethodsAn anonymous, internet-based, cross-sectional, open survey was distributed to medical and dental students at Fujian Medical University, Fuzhou, China. Data on the demographic information and characteristics of MSP were collected and analyzed. In addition to descriptive statistics, logistic regression was used to analyze significant risk factors contributing to MSP.ResultsA total of 1,178 students responded to the survey (response rate = 79.6%), including 722 medical students and 456 dental students. The age ranged from 16 to 24. There were 553 male students and 625 female students. A total of 523 students reported neck pain (NP, 44.4%), 361 students reported low back pain (LBP, 30.6%), and 182 students reported joint pain (JP, 15.4%). Sixty-six students (5.6%) simultaneously suffered from NP, LBP, and JP. The prevalence of NP (49.1 vs. 41.4%, P = 0.01), LBP (34.6 vs. 28.1%, P = 0.02), and JP (20.2 vs. 12.5%, P < 0.001) was significantly higher in dental students than in medical students. The prevalence of MSP was significantly different among the academic years for NP and LBP (P = 0.02 and P < 0.001, respectively). Univariate and multivariate regression analyses demonstrated that female sex, PSS-10 score, and major of stomatology were risk factors for MSP. Medical and dental students' intention to seek treatment for MSP was low and was significantly associated with the severity of pain.ConclusionsThe prevalence of MSP in Chinese medical and dental students is high, especially for NP and LBP, and is significantly higher in dental students than in medical students. The prevalence of NP and LBP were significantly different among academic grades. Female sex, PSS-10 score, and major of stomatology were risk factors for MSP. Students' intent to seek treatment for MSP was very low and was determined by the severity of pain.

BackgroundFungal osteoarticular infection (FOI) is not commonly seen in clinical practice but proposes a great challenge to orthopedic surgeons. In this study, we aimed to investigate the risk factors, the clinical features, and surgical outcomes of FOI in our institution. Specifically, we aimed to explore the role of metagenomic next-generation sequencing (mNGS) in the diagnosis and treatment of FOI.MethodsAll the patients who were diagnosed and managed with FOI in our institution from January 2007 to December 2020 were retrospectively reviewed, including primary fungal implant-related infection, primary fungal osteomyelitis or arthritis, and fungal infections secondary to bacterial osteomyelitis or implant-related bacterial infections. The potential risk factors and the clinical and surgical features were analyzed. The pathogen data were compared between culture and the mNGS test.ResultsA total of 25 patients were included, namely, 12 primary implant-related infections, 7 primary fungal osteomyelitis or arthritis, and 6 fungal infections secondary to bacterial osteomyelitis or implant-related bacterial infections. Most cases had undergone multiple surgeries or long-term antibiotic treatment. Diagnosis was mainly based on microbial culture and the mNGS test. Optimization of culture methods and the use of mNGS assisted the diagnosis. Specifically, mNGS was performed in 12 patients, 5 of whom were culture-negative. In the remaining seven cases, mNGS demonstrated the same results as culture. Management of FOI was complicated as most patients required multiple surgeries followed by long-term antifungal treatment. In selected cases, antifungal-impregnated cement spacer retention can be an optional choice. The overall success rate was 100% (25/25) for our cohort.ConclusionWe concluded that patients with comorbidities and a history of multiple surgeries or long-term antibiotics are under higher risk for FOI. Use of mNGS assists the diagnosis and treatment of FOI. Surgery combined with long-term antifungal treatment achieved satisfactory outcomes. In selected cases, antifungal-impregnated cement spacer retention can be an optional treatment choice.

Macrophage polarization followed by myocardial infarction (MI) is essential for wound healing. Tripartite motif-containing protein 21 (TRIM21), a member of E3 ubiquitin ligases, is emerging as a mediator in cardiac injury and heart failure. However, its function in modulating post-MI macrophage polarization remains elusive. Here, we detected that the levels of TRIM21 significantly increased in macrophages of wild-type (WT) mice after MI. In contrast, MI was ameliorated in TRIM21 knockout (TRIM21-/-) mice with improved cardiac remodeling, characterized by a marked decrease in mortality, decreased infarct size, and improved cardiac function compared with WT-MI mice. Notably, TRIM21 deficiency impeded the post-MI apoptosis and DNA damage in the hearts of mice. Consistently, the accumulation of M1 phenotype macrophages in the infarcted tissues was significantly reduced with TRIM21 deletion. Mechanistically, the deletion of TRIM21 orchestrated the process of M1 macrophage polarization at least partly via a PI3K/Akt signaling pathway. Overall, we identify TRIM21 drives the inflammatory response and cardiac remodeling by stimulating M1 macrophage polarization through a PI3K/Akt signaling pathway post-MI.

Escherichia coli has been engineered for L-malate production via aerobic cultivation. However, the maximum yield obtained through this mode is inferior to that of anaerobic fermentation due to massive amounts of CO2 emissions. Here, we aim to address this issue by reducing CO2 emissions of recombinant E. coli during aerobic L-malate production. Our findings indicated that NADH oxidation and ATP-synthesis-related genes were down-regulated with 2 g/L of YE during aerobic cultivations of E. coli E23, as compared to 5 g/L of YE. Then, E23 was engineered via the knockout of nuoA and the introduction of the nonoxidative glycolysis (NOG) pathway, resulting in a reduction of NAD+ and ATP supplies. The results demonstrate that E23 (ΔnuoA, NOG) exhibited decreased CO2 emissions, and it produced 21.3 g/L of L-malate from glucose aerobically with the improved yield of 0.43 g/g. This study suggests that a restricted NAD+ and ATP supply can prompt E. coli to engage in incomplete oxidization of glucose, leading to the accumulation of metabolites instead of utilizing them in cellular respiration.

Abstract Objectives We aimed to explore the bacterial community composition following ocean bacterial infection using an animal model. Methods This animal-based experiment was conducted from September 2019 to November 2019. Eighteen seawater filter membranes were collected from Changle City, Fujiian Province, China, on September 8, 2019. Ten filter membranes were used for implantation. Eight filter membranes that were used in the bacterial culture for the exploration of seawater bacteria were assigned to the seawater group (SG). Fourteen healthy adult New Zealand rabbits were randomly divided into the experimental group (EG) and control group (CG). Seawater filter membranes and asepsis membranes were implanted into the tibia in the EG and CG, respectively. One week after surgery, tibial bone pathology tissues were collected and assessed using light microscopy and scanning electron microscopy (SEM). Medullary cavity tissues were collected for the performance of Illumina MiSeq sequencing and bacterial culture. The differences between EG and CG were assessed by pathological observation under light microscopy and SEM, high-throughput bacterial sequencing, and bacterial culture. Results Compared with the CG, the infection rate was 100%, and the mortality value was 20% after the implantation of the filter membranes in the EG. Both light microscopy and SEM showed that a large number of bacteria were distributed in the bone marrow cavity after ocean bacterial infection. No bacterial growth was found in the CG. Illumina MiSeq sequencing found that Firmicutes, Proteobacteria, Thermotogae, Fusobacteria, Bacteroidetes, and Actinobacteria were the dominant bacteria at the phylum level and Clostridium_sensu_stricto_7, Haloimpatiens, Clostridium_sensu_stricto_15, Clostridiaceae_1, Clostridium_sensu_stricto_18, and Oceanotoga were the dominant bacteria in genus level among the EG. In the bacterial culture of the medullary cavity tissues, Klebsiella pneumoniae, Shewanella algae, Staphylococcus aureus, Escherichia coli, Enterobacter cloacae, and Vibrio vulnificus were the predominant infective species. Moreover, compared with the SG, the EG showed a higher detection rate of E. coli and S. aureus (P = 0.008 and P = 0.001, respectively). The detection rates of V. alginolyticus, V. parahaemolyticus, and V. fluvialis were higher in the SG than the EG (P = 0.007, P = 0.03, and P = 0.03, respectively). Conclusions Our model, which was comprehensively evaluated using four techniques: histopathology and SEM observation, gene detection, and bacteria culture, provides a scientific basis for the clinical diagnosis and treatment of patients in such settings.

Methanol is an attractive C1 feedstock with high abundance and low cost in bio-manufacturing. However, the metabolic construction of cell factories to utilize methanol for chemicals production remains a challenge due to the toxic intermediates and complicated metabolic pathways. The group of Zhou rescued methylotrophic yeast from cell death and achieved high-level production of free fatty acids from methanol through a combination of adaptive laboratory evolution, rational metabolic engineering and multi-omics analysis.

The economic and cultural effects of sports films have attracted close attention from academia as well as the industry. In this paper, two sub-studies were conducted to explore the empathy mechanism performance of the interest-related community in sports films. In Study 1, the film Lead was applied as an example and used network text analysis to analyze the discourse characteristics and structure of its interest-related community to grasp the practice regularities. More specifically, the results in Study 1 show that the theme feature, brand building, technological innovation, and spirit communication are the key factors that affect an individual's empathy for sports films. In Study 2, I conducted a survey to explore the empathy mechanism of sports films in the new age to provide a theoretical reference for the high-quality development of films. The above-mentioned four main factors have positive impacts on empathy: affective empathy and cognitive empathy.

Abstract Background In view of the ongoing coronavirus disease (COVID-19) pandemic, it remains unclear whether the severity of illness and time interval from symptom onset to release from quarantine differ between cases that originated from clusters and cases reported in other areas. This study aimed to assess epidemiological and intergenerational clinical characteristics of COVID-19 patients associated with cluster outbreaks to provide valuable data for the prevention and control of COVID-19. Methods We identified the first employee with COVID-19 at a supermarket and screened the close contacts of this index patient. Confirmed cases were divided into two groups according to the generation (first generation comprising supermarket employees [group A] and second or third generations comprising family members or friends of the supermarket employees [group B]). The epidemiological and clinical characteristics of the two groups were retrospectively compared. Results A total of 8437 people were screened, and 24 COVID-19 patients were identified. Seven patients (29.2%) were asymptomatic; three patients were responsible for six symptomatic cases. The interval from the confirmation of the first case to symptom onset in symptomatic patients was 5–11 days. The clinical manifestations of symptomatic patients upon admission were non-specific. All patients (including the seven asymptomatic patients) were admitted based on chest computed tomography features indicative of pneumonia. There were 11 cases in group A (first generation) and 13 cases in group B (second generation, 11 cases; third generation, 2 cases), with no significant differences in clinical and epidemiological characteristics between the two groups, except for sex, duration from symptom onset to hospitalization, and underlying disease (P > 0.05). Conclusions For cluster outbreaks, it is important to comprehensively screen close the contacts of the index patient. Special attention should be paid to asymptomatic cases. The clinical management of cluster patients is similar to that of other COVID-19 patients.

The topology structure of multi-layer networks is highly correlated with the robustness of consensus. This paper investigates the influence of different interlayer edge connection patterns on the consensus of the two-layer ring networks. Two types of two-layer ring network models are first considered: one is a kind of two-layer ring network with two linked edges between layers (Networks-a), and the other is a kind of two-layer ring network with three linked edges between layers (Networks-b). Using the Laplacian spectrum, the consensus of the network model is derived. The simulation experiments are used to demonstrate the influence of different interlayer edge connection patterns on the consensus of networks. To determine the best edge connection pattern for Networks-a and Networks-b, the number of nodes in a single-layer ring network is denoted by n. The best edge connection pattern for Networks-a is 1 & [(n+2)/2]. Furthermore, n is subdivided into 3k,3k+1,3k+2, and the best edge connection patterns of Networks-b are near 1 & k+1 & 2k+1.

This paper proposes an integrated path tracking controller for articulated vehicles. A nonlinear model-predictive control (NMPC)-based reference state tracker is designed as an upper-level controller to solve the vehicle’s longitudinal velocity and steering rate. A terminal cost is introduced into the NMPC to improve the controller’s stability. A lower-level controller is developed to translate upper-level solutions into vehicle actuators’ signals, including steering and driving controllers. The steering controller translates the steering rate into the linear velocity of the cylinder to calculate the required fluid volume and ultimately into the rotation speed of the steering motor. The neural network method is applied in the driving controller to ensure accuracy under different loadings. In order to investigate the effects of the path tracking controller, an articulated dump truck is adapted for the field tests by adding the steering-by-wire system and driving-by-wire system, respectively. Experimental verifications of the lower-level controller are performed. The results show that the controller can accurately satisfy the demand. Finally, the tracking performance of the integrated path tracking controller is analyzed experimentally under different reference velocities. The results indicate that tracking accuracy can be guaranteed.

Recent developments in ultrafine bubble generation have opened up new possibilities for applications in various fields. Herein, we investigated how substances in water affect the size distribution and stability of microbubbles generated by a common nanobubble generator. By combining light scattering techniques with optical microscopy and high-speed imaging, we were able to track the evolution of microbubbles over time during and after bubble generation. Our results showed that air injection generated a higher number of microbubbles (μm) than CO2 injection. Increasing detergent concentration led to a rapid increase in the number of microbubbles generated by both air and CO2 injection and the intensity signal detected by dynamic light scattering (DLS) slightly increased. This suggested that surface-active molecules may inhibit the growth and coalescence of bubbles. In contrast, we found that salts (NaCl and Na2CO3) in water did not significantly affect the number or size distribution of bubbles. Interestingly, the presence of oil in water increased the intensity signal and we observed that the bubbles were coated with an oil layer. This may contribute to the stability of bubbles. Overall, our study sheds light on the effects of common impurities on bubble generation and provides insights for analyzing dispersed bubbles in bulk.

Vanillin has been widely used as a flavoring agent in the food industry and as a precursor in the medicine and polymer industries. However, the use of chemically synthesized vanillin is prohibited in food and some other industries. Additionally, the harsh conditions and toxic substrates in chemically synthesized vanillin lead to some environmental challenges and energy waste. With the rapid development of synthetic biology, the biological production of vanillin from renewable resources through microbial fermentation has gained great attention owing to its high selectivity and environmentally friendly properties. Accordingly, this article will discuss the vanillin biosynthesis technology from the aspects of chassis cell types and substrate types. The key enzymes involved in metabolic pathways are also discussed. Then, we summarize some improvements in the process of vanillin production to increase its production and reduce the toxicity of vanillin in microorganisms, and the possible future directions for vanillin biosynthesis will also be outlined.

Abstract Background Gastric ectopic pancreas (GEPs) is a rare developmental anomaly which is difficult to differentiate it from submucosal tumor such as gastric stromal tumor (GST) by imaging methods. Since the treatments of the GEPs and GST are totally different, a correct diagnosis is essential. Therefore, we retrospectively investigated the CT features of them to help us deepen the understanding of GEPs and GST. Methods This study enrolled 17 GEPs and 119 GST, which were proven pathologically. We assessed clinical and CT features to identify significant differential features of GEPs from GST using univariate and multivariate analyses. Results In univariate analysis, among all clinicoradiologic features, features of age, symptom, tumor marker, location, contour, peritumoral infiltration or fat-line of peritumor, necrosis, calcification, CT attenuation value of unenhancement phase/arterial phase/portal venous phase (CTu/CTa/CTp), the CT attenuation value of arterial phase/portal venous phase minus that of unenhanced phase (DEAP/DEPP), long diameter (LD), short diameter (SD) were considered statistically significant for the differentiation of them. And the multivariate analysis revealed that location, peritumoral infiltration or fat-line of peritumor, necrosis and DEPP were independent factors affecting the identification of them. In addition, ROC analysis showed that the test efficiency of CTp was perfect (AUC = 0.900). Conclusion Location, the presence of peritumoral infiltration or fat-line of peritumor, necrosis and DEPP are useful CT differentiators of GEPs from GST. In addition, the test efficiency of CTp in differentiating them was perfect (AUC = 0.900).

Abstract Background This study aimed to evaluate the effects of different pretreatment methods on the microbial yield from infectious tissues. Methods Strains of Staphylococcus aureus (SA), Escherichia coli (EC) and Candida albicans (CA) were used to construct single-surface, full-surface, and internal infection models in sterile pork tissue. Manual milling (MM), mechanical homogenization (MH), sonificated (SF), dithiothreitol (DTT), and direct culture (DC) were used to pretreat these tissues, the microbial yield from different pretreatment methods were recorded and compared. Moreover, periprosthetic tissues collected intraoperatively from periprosthetic joint infection (PJI) patients were used as a verification. Results The study showed that the microbial yield from MH pretreatment was significantly higher than that of MM (P

Aims: Microbiological culture is a key element in the diagnosis of periprosthetic joint infection (PJI). However, cultures of periprosthetic tissue do not have optimal sensitivity. One of the main reasons for this is that microorganisms are not released from the tissues, either due to biofilm formation or intracellular persistence. This study aimed to optimize tissue pretreatment methods in order to improve detection of microorganisms. Methods: From December 2017 to September 2019, patients undergoing revision arthroplasty in a single centre due to PJI and aseptic failure (AF) were included, with demographic data and laboratory test results recorded prospectively. Periprosthetic tissue samples were collected intraoperatively and assigned to tissue-mechanical homogenization (T-MH), tissue-manual milling (T-MM), tissue-dithiothreitol (T-DTT) treatment, tissue-sonication (T-S), and tissue-direct culture (T-D). The yield of the microbial cultures was then analyzed. Results: A total of 46 patients were enrolled, including 28 patients in the PJI group and 18 patients in the AF group. In the PJI group, 23 cases had positive culture results via T-MH, 22 cases via T-DTT, 20 cases via T-S, 15 cases via T-MM, and 13 cases via T-D. Three cases under ongoing antibiotic treatment remained culture-negative. Five tissue samples provided the optimal yield. Any ongoing antibiotic treatment had a relevant influence on culture sensitivity, except for T-DTT. Conclusion: T-MH had the highest sensitivity. Combining T-MH with T-DTT, which requires no special equipment, may effectively improve bacterial detection in PJI. A total of five periprosthetic tissue biopsies should be sampled in revision arthroplasty for optimal detection of PJI.

Abstract Clostridium sp. strain CT7 is a new emerging microbial cell factory with high butanol production ratio owing to its non-traditional butanol fermentation mode with uncoupled acetone and 1,3-propanediol formation. Significant changes of metabolic products profile were shown in glycerol- and glucose-fed strain CT7, especially higher butanol and lower volatile fatty acids (VFAs) production occurred from glycerol-fed one. However, the mechanism of this interesting phenomenon was still unclear. To better elaborate the bacterial response towards glycerol and glucose, the quantitative proteomic analysis through iTRAQ strategy was performed to reveal the regulated proteomic expression levels under different substrates. Proteomics data showed that proteomic expression levels related with carbon metabolism and solvent generation under glycerol media were highly increased. In addition, the up-regulation of hydrogenases, ferredoxins and electron-transferring proteins may attribute to the internal redox balance, while the earlier triggered sporulation response in glycerol-fed media may be associated with the higher butanol production. This study will pave the way for metabolic engineering of other industrial microorganisms to obtain efficient butanol production from glycerol.

Abstract Background The global production of glycerol is increasing year by year since the demands of biodiesel is rising. It is benefit for high-yield succinate synthesis due to its high reducing property. A. succinogenes, a succinate-producing candidate, cannot grow on glycerol anaerobically, as it needs a terminal electron acceptor to maintain the balance of intracellular NADH and NAD+. Microbial fuel cell (MFC) has been widely used to release extra intracellular electrons. However, A. succinogenes is a non-electroactive strain which need the support of electron shuttle in MFC, and pervious research showed that acid-tolerant A. succinogenes has higher content of unsaturated fatty acids, which may be beneficial for the transmembrane transport of lipophilic electron shuttle. Results MFC-assisted succinate production was evaluated using neutral red as an electron shuttle to recover the glycerol utilization. First, an acid-tolerant mutant JF1315 was selected by atmospheric and room temperature plasma (ARTP) mutagenesis aiming to improve transmembrane transport of neutral red (NR). Additionally, MFC was established to increase the ratio of oxidized NR to reduced NR. By combining these two strategies, ability of JF1315 for glycerol utilization was significantly enhanced, and 23.92 g/L succinate was accumulated with a yield of 0.88 g/g from around 30 g/L initial glycerol, along with an output voltage above 300 mV. Conclusions A novel MFC-assisted system was established to improve glycerol utilization by A. succinogenes for succinate and electricity production, making this system as a platform for chemicals production and electrical supply simultaneously.

Osteoarthritis (OA) is a debilitating joint disease that affects millions of individuals. The pathogenesis of OA has not been fully elucidated. Obesity is a well-recognized risk factor for OA. Multiple studies have demonstrated adipokines play a key role in obesity-induced OA. Increasing evidence show that various adipokines may significantly affect the development or clinical course of OA by regulating the pro/anti-inflammatory and anabolic/catabolic balance, matrix remodeling, chondrocyte apoptosis and autophagy, and subchondral bone sclerosis. Several signaling pathways are involved but still have not been systematically investigated. In this article, we review the cellular and molecular mechanisms of adipokines in OA, and highlight the possible signaling pathways. The review suggested adipokines play important roles in obesity-induced OA, and exert downstream function via the activation of various signaling pathways. In addition, some pharmaceuticals targeting these pathways have been applied into ongoing clinical trials and showed encouraging results. However, these signaling pathways are complex and converge into a common network with each other. In the future work, more research is warranted to further investigate how this network works. Moreover, more high quality randomised controlled trials are needed in order to investigate the therapeutic effects of pharmaceuticals against these pathways for the treatment of OA. This review may help researchers to better understand the pathogenesis of OA, so as to provide new insight for future clinical practices and translational research.

Objectives: Preoperative diagnosis is important for patients who need revision surgery due to PJI. Microbial culture plays an important role in PJI diagnosis, but the sensitivity of cultures is low when the sample amount is limited or when a patient is treated with antibiotics before sample collection. In this study, metagenomic next-generation sequencing (mNGS) was used to detect bacteria in preoperative puncture synovial fluid samples from patients with suspected PJI, and the preoperative and intraoperative culture results were compared to estimate its diagnostic efficiency. Methods: From July 2016 to December 2018, patients with suspected PJI who underwent prosthetic joint revision surgery were included, and the results of those who had been tested by preoperative synovial fluid culture and mNGS were obtained. The demographic characteristics, medical history, laboratory test results, culture results, and mNGS results of each patient were recorded. Then, the efficiency of preoperative synovial fluid mNGS was compared to that of synovial fluid culture for diagnosing PJI. Results: A total of 37 patients were included, and 24 patients (25 joints) were diagnosed with PJI. The sensitivity, specificity, and accuracy of preoperative synovial fluid mNGS were 92%, 91.7%, and 83.7%, respectively. The sensitivity, specificity, and accuracy of preoperative synovial fluid culture were 52%, 91.7%, and 43.7%, respectively. Conclusions: With a low volume of synovia (1 ml), mNGS can be performed with higher sensitivity and specificity compared to synovial culture. Thus, mNGS can be a useful supplemental method to improve diagnostic efficiency during the preoperative period.

Abstract Background l-malate is one of the most important platform chemicals widely used in food, metal cleaning, textile finishing, pharmaceuticals, and synthesis of various fine chemicals. Recently, the development of biotechnological routes to produce l-malate from renewable resources has attracted significant attention. Results A potential l-malate producing strain E. coli BA040 was obtained by inactivating the genes of fumB, frdABCD, ldhA and pflB. After co-overexpression of mdh and pck, BA063 achieved 18 g/L glucose consumption, leading to an increase in l-malate titer and yield of 13.14 g/L and 0.73 g/g, respectively. Meantime, NADH/NAD+ ratio decreased to 0.72 with the total NAD(H) of 38.85 µmol/g DCW, and ATP concentration reached 715.79 nmol/g DCW. During fermentation in 5L fermentor with BA063, 41.50 g/L glucose was consumed within 67 h with the final l-malate concentration and yield of 28.50 g/L, 0.69 g/g when heterologous CO2 source was supplied. Conclusions The availability of NAD(H) was correlated positively with the glucose utilization rate and cellular metabolism capacities, and lower NADH/NAD+ ratio was beneficial for the accumulation of l-malate under anaerobic conditions. Enhanced ATP level could significantly enlarge the intracellular NAD(H) pool under anaerobic condition. Moreover, there might be an inflection point, that is, the increase of NAD(H) pool before the inflection point is followed by the improvement of metabolic performance, while the increase of NAD(H) pool after the inflection point has no significant impacts and NADH/NAD+ ratio would dominate the metabolic flux. This study is a typical case of anaerobic organic acid fermentation, and demonstrated that ATP level, NAD(H) pool and NADH/NAD+ ratio are three important regulatory parameters during the anaerobic production of l-malate.

Objectives: Pathogen identification is crucial for the diagnosis and management of periprosthetic joint infection (PJI). Although culturing methods are the foundation of pathogen detection in PJI, false-negative results often occur. Oxford nanopore sequencing (ONS) is a promising alternative for detecting pathogens and providing information on their antimicrobial resistance (AMR) profiles, without culturing. Methods: To evaluate the capability of metagenomic ONS (mONS) in detecting pathogens from PJI samples, both metagenomic next-generation sequencing (mNGS) and mONS were performed in 15 osteoarticular samples from nine consecutive PJI patients according to the modified Musculoskeletal Infection Society (MSIS) criteria. The sequencing data generated from both platforms were then analyzed for pathogen identification and AMR detection using an in-house-developed bioinformatics pipeline. Results: Our results showed that mONS could be applied to detect the causative pathogen and characterize its AMR features in fresh PJI samples. By real-time sequencing and analysis, pathogen identification and AMR detection from the initiation of sequencing were accelerated. Conclusion: We showed proof of concept that mONS can function as a rapid, accurate tool in PJI diagnostic microbiology. Despite efforts to reduce host DNA, the high proportion of host DNA was still a limitation of this method that prevented full genome analysis.

Objectives: The purpose of this study was to explore the role of mNGS in the diagnosis of PJI by detecting the presence of bacteria in periprosthetic tissues. Methods: Patients who were suspected of PJI and underwent surgery in our center were enrolled. Microbial culture and mNGS were performed on periprosthetic tissues collected intraoperatively. Patient age, body mass index (BMI), laboratory tests, microbial culture and mNGS results were recorded. The diagnostic performance of microbial culture and mNGS in the diagnosis of PJI was compared. Results: A total of 44 patients were included. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of periprosthetic tissue microbial culture in the diagnosis of PJI were 72.72%, 77.27%, 76.19%, 73.91% and 75%, respectively. The sensitivity, specificity, PPV, NPV and accuracy of periprosthetic tissue mNGS in the diagnosis of PJI were 95.45%, 90.91%, 91.3%, 95.24% and 93.18%, respectively. The sensitivity and specificity of mNGS in the diagnosis of PJI were superior to those of microbial culture. Conclusion: Periprosthetic tissue can be used as an alternative to synovial fluid and sonication fluid, as mNGS can improve the diagnosis of PJI. mNGS of the periprosthetic tissues showed higher sensitivity than microbial culture.

Abstract Background Oxford unicompartmental knee arthroplasty (UKA) is widely used for treating patients with unicompartmental knee diseases. However, the factors affecting the outcomes of and patients’ satisfaction with Oxford UKA remain controversial. The aims of this study were to evaluate the clinical and radiological outcomes, including patient satisfaction, after Oxford UKA and identify the influencing factors related to patients’ satisfaction. Methods We retrospectively analyzed the data of patients who underwent UKA in our medical institution from 2013 to 2018. Demographic information, clinical scores, patient satisfaction and imaging findings were recorded. The patients were followed up for at least 1 year. Multivariate Logistic regression analysis was performed to identify influencing factors related to clinical outcomes including patient satisfaction. Results A total of 80 patients (involving 87 knees) were included. The Numerical Rating Scale (NRS) scores and Knee Society Scores (KSS) were significantly improved after operation as compared to preoperative scores (6.0 vs. 1.0, P

Purpose: The aims of our study were to (1) evaluate the concordance of both methods for detecting prosthetic joint infection (PJI) pathogens in joint fluid and to (2) clarify whether broad-range polymerase chain reaction (BR-PCR) can be used as a verification method for metagenomic next-generation sequencing (mNGS) for PJI diagnosis. Methods: In total, 63 patients underwent total joint arthroplasty, with 45 PJI and 18 aseptic failure patients included. Joint fluids were sampled after antibiotics were withheld for more than 2 weeks, and then, culture, BR-PCR and mNGS were performed for all samples. Results: The joint fluid BR-PCR sensitivity was 82.2%, which was not significantly different from that of mNGS (95.6%) or culture (77.8%). The specificities of the 3 methods were all 94.4%. BR-PCR failed to identify the pathogens in 1 polymicrobial infection patient and 4 fungal infection patients. Conclusion: mNGS was more sensitive than BR-PCR for detecting PJI pathogens in joint fluid. BR-PCR is insufficient for use as an mNGS verification method.

Abstract Background Pulmonary Cryptococcosis is a common fungal infection mainly caused by Cryptococcus neoformans/C.gattii species in immunocompromised patients. Cases of pulmonary cryptococcosis in patients with normal immune function are increasingly common in China. Clinical and radiographic features of pulmonary cryptococcosis are various and without obvious characteristics, so it is often misdiagnosed as pulmonary metastatic tumor or tuberculosis. When coexisting with malignant lung tumors, it was more difficult to differentiate from metastatic lung cancer, although the coexistence of pulmonary cryptococcosis and central type lung cancer is rare. Reviewing the imaging manifestations and diagnosis of the case and the relevant literature will contribute to recognition of the disease and a decrease in misdiagnoses. Case presentation A 72-year-old immunocompetent Han Chinese man had repeated dry cough for more than half a year. CT examination of chest showed an irregular mass at the left hilum of the lung, and two small nodules in the right lung, which were considered as the left central lung cancer with right lung metastasis. However, the patient was diagnosed with pulmonary cryptococcosis coexisting with central type lung cancer based on the results of laboratory examination, percutaneous lung biopsy, fiberoptic bronchoscopy, and surgical pathology. The patient underwent surgical resection of the left central type lung cancer and was placed on fluconazole treatment after a positive diagnosis was made. Five years after the lung cancer surgery, the patient had a recurrence, but the pulmonary cryptococcus nodule disappeared. Conclusion Our case shows that CT findings of central type lung cancer with multiple pulmonary nodules are not necessarily metastases, but may be coexisting pulmonary cryptococcosis. CT images of cryptococcosis of the lung were diverse and have no obvious characteristics, so it was very difficult to distinguish from metastatic tumors. CT-guided percutaneous lung biopsy was a simple and efficient method for identification.

Abstract Background Rhamnolipids are the best known microbial-derived biosurfactants, which has attracted great interest as potential ‘‘green” alternative for synthetic surfactants. However, rhamnolipids are the major contributors to severe foam problems, which greatly inhibit the economics of industrial-scale production. In this study, a novel foam-control system was established for ex situ dealing with the massive overflowing foam. Based on the designed facility, foam reduction efficiency, rhamnolipids production by batch and repeated fed-batch fermentation were comprehensively investigated. Results An ex situ foam-control system was developed to control the massive overflowing foam and improve rhamnolipids production. It was found that the size of individual bubble in the early stage was much larger than that of late fermentation stage. The foam liquefaction efficiency decreased from 54.37% at the beginning to only 9.23% at the end of the fermentation. This difference of bubble stability directly resulted in higher foam reduction efficiency of 67.46% in the early stage, whereas the small uniform bubbles can only be reduced by 57.53% at the later fermentation stage. Moreover, reduction of secondary foam is very important for foam controlling. Two improved designs of the device in this study obtained about 20% improvement of foam reduction efficiency, respectively. The batch fermentation result showed that the average volume of the overflowing foam was reduced from 58–640 to 19–216 mL/min during the fermentation process, presenting a notable reduction efficiency ranging from 51.92 to 73.47%. Meanwhile, rhamnolipids production of batch fermentation reached 45.63 g/L, and the yield 0.76 g/g was significantly better than ever reported. Further, a repeated fed-batch fermentation based on the overall optimization was carried out. Total rhamnolipids concentration reached 48.67 g/L with the yield around of 0.67–0.83 g/g, which presented an improvement of 62% and 49% compared with conventional batch fermentation by using various kinds of defoamers, respectively. Conclusions The ex situ foam-control system presented a notable reduction efficiency, which helped greatly to easily solve the severe foaming problem without any defoamer addition. Moreover, rhamnolipids production and yield by repeated fed-batch fermentation obtained prominent improvement compared to conventional batch cultivation, which can further facilitate economical rhamnolipids production at large scales.

Abstract Spinal osteosarcoma (OS) is a rare and aggressive malignancy. Long noncoding RNA (lncRNA) BSN-AS2 has been shown to be an oncogenic gene in several cancers. However, the role and function of BSN-AS2 in spinal OS were unfamiliar. Our study identified that BSN-AS2 expression was boosted in spinal OS tissues and cell lines. Transcription factor E2F1 induced the upregulation of BSN-AS2 expression in spinal OS cells. Afterwards, loss-of-function assays indicated that BSN-AS2 depletion reduced cell proliferation, migration and invasion as well as promoted cell apoptosis in spinal OS. Thereafter, RIP, RNA pull down and luciferase reporter assays manifested BSN-AS2 could sponge miR-654-3p in spinal OS. After that, the binding effect of between miR-654-3p and SYTL2 was proved. Finally, rescue experiments illustrated that miR-654-3p inhibition or SYTL2 overexpression could counteract the inhibitory effect caused by BSN-AS2 deficiency on spinal OS progression. In conclusion, the availability of miR-654-3p was antagonized by E2F1-induced BSN-AS2 for SYTL2-meidated spinal OS progression.

Abstract Background The aims of this study were to (1) evaluate the efficacy and safety of targeted antibiotics for the treatment of culture-negative prosthetic joint infection based on metagenomic next-generation sequencing results and (2) verify the accuracy and reliability of metagenomic next-generation sequencing for identifying pathogens related to culture-negative prosthetic joint infection. Methods Ninety-seven consecutive PJI patients, including 27 patients with culture-negative prosthetic joint infection, were treated surgically at our center. Thirteen of the 27 culture-negative prosthetic joint infection patients, who were admitted before June 2017 and treated with empirical antibiotics, comprised the empirical antibiotic group (EA group), and the other 14 patients, who were admitted after June 2017 and treated with targeted antibiotics according to their metagenomic next-generation sequencing results, were classified as the targeted antibiotic group (TA group). The short-term infection control rate, incidence of antibiotic-related complications and costs were compared between the two groups. Results Two of the patients in the EA group experienced debridement and prolonged antimicrobial therapy due to wound infection after the initial revision surgery. No recurrent infections were observed in the TA group; however, no significant difference in the infection control rate was found between the two groups (83.33% vs 100%, P = 0.217). More cases of antibiotic-related complications were recorded in the EA group (6 cases) than in the TA group (1 case), but the difference was not statistically significant (P = 0.0697). The cost of antibiotics obtained for the EA group was 20,168.37 Yuan (3236.38–45,297.16), which was higher than that found for the TA group (10,164.16 Yuan, 2959.54–16,661.04, P = 0.04). Conclusions Targeted antibiotic treatment for culture-negative prosthetic joint infection based on metagenomic next-generation sequencing results is associated with a favorable outcome, and metagenomic next-generation sequencing is a reliable tool for identifying pathogens related to culture-negative prosthetic joint infection.

Structural thermal deformation is an important factor that affects the performance of MEMS devices. The mismatch of thermal expansion coefficient (CTE) between different materials is a major source. For single center supported MEMS devices, expansion difference between device and the substrate leads to the out of plane thermal deformation of the structure, resulting in the performance deterioration. This paper presents a thermal deformation suppression chip (TDSC) for single center supporting MEMS devices. It is fabricated by the MEMS process and consists of an upper plate and a lower plate. The materials of the upper and lower plates are the same as those of the device substrate and structure respectively. The principle of TDSC to suppress thermal deformation is material symmetry design. Single center anchor is adapted to connect the upper and lower plates. Besides, the center anchor can also isolate the packaging stress generated between TDSC and package shell. In this paper, center supported quadruple mass gyroscope (CSQMG) is used to verify the effect of TDSC. Finite element simulation shows that the thermal deformation suppression effect is determined by radius of TDSC anchor, and the deformation can be suppressed to 0. The conclusion is confirmed by White light interference experiment. In addition, the experiment results also show that the TDSC effectively reduce the thermal out-of-plane deformation after packaging, which is even better than that of the gyroscope without any packaging. It is further proved that TDSC can also significantly improve the temperature performance of the CSQMG. Moreover, the TDSC has simple process steps, small chip area, low cost, and is suitable for various MEMS devices with single center anchor.

ABSTRACT: This experiment was conducted to investigate the effects of dietary supplementation with Bacillus amyloliquefaciens CECT 5940 (BA-5940) on growth performance, antioxidant capacity, immunity, and digestive enzyme activity of broiler chickens. A total of 720 one-day-old Arbor Acres male broiler chicks (average body weight, 45.87 ± 0.86 g) were randomly allocated to 5 treatments of 8 replicates with 18 chicks in each replicate. Broilers in the control group were fed a corn-wheat-soybean basal diet, the other 4 groups were fed the same basal diet supplemented with 500, 1,000, 1,500, or 2,000 mg/kg Ecobiol (1.27 × 109 CFU/g BA-5940) for 42 d, respectively. Broilers fed diets supplemented with BA-5940 showed a quadratic response (P < 0.05) of average daily gain (ADG) and feed to gain ratio (F:G) during d 22 to 42 and d 0 to 42. The glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities in serum and liver, and total antioxidant capacity (T-AOC) in liver of broilers on d 42 increased linearly (P < 0.05) with increasing levels of BA-5940, while malondialdehyde (MDA) level in serum decreased linearly (P < 0.05). Concentrations of serum immunoglobulin (Ig) A and IgM on d 21, and IgM on d 42 increased linearly (P < 0.05) as BA-5940 levels increased. Supplementation with increasing levels of BA-5940 linearly decreased serum tumor necrosis factor-α (TNF-α) levels on d 21 and 42, while increased interleukin (IL)-10 concentration (linear, P < 0.05) on d 21. Meanwhile, the levels of IL-1β, IL-6, and TNF-α in the mucosa of jejunum and ileum were decreased (linear, P < 0.05) on d 42 as dietary supplementation of BA-5940 increased. Additionally, supplementation with BA-5940 also increased the activities of amylase (linear, P < 0.01), lipase (linear, P < 0.05) and chymotrypsin (linear, P < 0.01) in jejunal digesta, and lipase (linear, P < 0.05) in ileal digesta of broilers on d 42. To summarize, inclusion of BA-5940 in corn-wheat-soybean meal-based diet improved growth performance of broilers through improving antioxidant capacity, immunity, and digestive enzyme activity. Based on the results of this study, 1.1–1.6 × 109 CFU/kg BA-5940 is recommended for supplementation in broiler diets.

The present study determined the effects of different probiotic mixture supplementation to sows from late pregnancy to day 21 postpartum on reproductive performance, colostrum composition, plasma biochemical parameters, and fecal microbiota and metabolites. A total of 80 pregnant sows were randomly assigned to one of four groups (20 sows per group). The sows in the control group (CON group) were fed a basal diet, and those in the BS-A+B, BS-A+BL, and BS-B+BL groups were fed basal diets supplemented with 250 g/t of different probiotic mixture containing either 125 g/t of Bacillus subtilis A (BS-A), Bacillus subtilis B (BS-B), and/or Bacillus licheniformis (BL), respectively. The trial period was from day 85 of pregnancy to day 21 postpartum. The results showed that different dietary probiotic mixture supplementation increased (P < 0.05) the average weaning weight and average daily gain of piglets, while dietary BS-A+BL supplementation increased the number of weaned piglets (P < 0.05), litter weight (P = 0.06), litter weight gain (P = 0.06), and litter daily gain (P = 0.06) at weaning compared with the CON group. Different dietary probiotic mixture supplementation improved (P < 0.05) the colostrum quality by increasing the fat and dry matter concentrations, as well as the protein and urea nitrogen concentrations in the BS-A+BL group. Dietary probiotic mixture BS-B+BL increased the plasma total protein on days 1 and 21 postpartum while decreased the plasma albumin on day 1 postpartum (P < 0.05). In addition, the plasma high-density lipoprotein-cholesterol was increased in the BS-A+B and BS-B+BL groups on day 21 postpartum, while plasma ammonia was decreased in the BS-A+B and BS-A+BL groups on day 1 and in the three probiotic mixtures groups on day 21 postpartum (P < 0.05). Dietary supplementation with different probiotic mixture also modified the fecal microbiota composition and metabolic activity in sows during pregnancy and postpartum stages. Collectively, these findings suggest that maternal supplementation with Bacillus subtilis in combination with Bacillus licheniformis are promising strategies for improving the reproductive performance and the overall health indicators in sows, as well as the growth of their offspring.

Purpose: This study aimed to investigate the potential mechanisms and related bioactive components of ZSS for the treatment of insomnia.Method: The insomnia model of rat induced by PCPA was established. After oral administration of ZSS extract, the general morphological observation, pentobarbital sodium-induced sleep test and histopathological evaluation were carried out. Network pharmacology, assisted by UHPLC-Q-Exactive-MS/MS analysis, was developed to identify the targets of ZSS in the treatment of insomnia, as well as the corresponding signaling pathways. In addition, we validated the identified targets and pathways by RT-qPCR and immunohistochemical analysis.Results: The pentobarbital sodium-induced sleep test, determination of 5-HT and GABA levles in hypothalamic tissues and HE staining showed that ZSS extract was an effective treatment for insomnia. Network pharmacology analysis identified a total of 19 candidate bioactive ingredients in ZSS extract, along with 433 potentially related targets. Next, we performed protein-protein interaction (PPI), MCODE clustering analysis, GO functional enrichment analysis, KEGG pathway enrichment analysis, and ingredient-target-pathway (I-T-P) sub-networks analysis. These methods allowed us to investigate the synergistic therapeutic effects of crucial pathways, including the serotonergic and GABAergic synapse pathways. Our analyses revealed that palmitic acid, coclaurine, jujuboside A, N-nornuciferine, caaverine, magnoflorine, jujuboside B, and betulinic acid, all played key roles in the regulation of these crucial pathways. Finally, we used the PCPA-induced insomnia in rats to validate the data generated by network pharmacology; these in vivo experiments clearly showed that pathways associated with the serotonergic and GABAergic system were activated in the rats model. Furthermore, ZSS treatment significantly suppressed high levels of HTR1A, GABRA1, and GABRG2 expression in the hypothalamus and reduced the expression levels of HTR2A.Conclusion: Based on the combination of comprehensive network pharmacology and in vivo experiments, we successfully identified the potential pharmacological mechanisms underlying the action of ZSS in the treatment of insomnia. The results provide a theoretical basis for further development and utilization of ZSS, and also provide support for the development of innovative drugs for the treatment of insomnia.