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Summary: The microbiota-gut-brain axis (MGBA) plays a pivotal role in drug addiction. However, the pathophysiological mechanism of MGBA in ketamine addiction remains elusive. The present study investigated the ketamine-induced gut microbiota disorders, intestinal barrier dysfunction, and the alterations in brain function, using a conditioned place preference (CPP) model of ketamine addiction in rats. Compared with the control group, ketamine induced decreased amplitude of low-frequency fluctuation (ALFF) values in the hippocampus, and pyknotic nuclei and concentrated cytoplasm in hippocampal neurons, as well as alterations in gut microbiota composition, shortened ileum villi, and thinner colonic mucosa. We also found that the abundance of gut microbiota exhibited correlations with CPP score, hippocampal ALFF value, length of ileum villi, and thickness of colonic mucosa. Our findings provide evidence for abnormal alterations in the MGBA of ketamine-addicted rats, which improves our understating of the mechanism of ketamine addiction and the potential for developing new therapeutic strategies.

Background: Angrogenetic alopecia (AGA) is one of the most prevalent hair loss disorders worldwide. The hair follicle stem cell (HFSC) is closely related to the formation of hair follicle (HF) structure and HF self-renewal. The activation of HFSC in AGA is critical for hair growth. Pilose antler has been reported to have hair growth-promoting activity, but the mechanism of action on AGA and HFSC has not been reported.Methods: We previously extracted an active component from the pilose antler known as PAEs. In this study, we conducted experiments using AGA mice and HFSC. The effects of PAEs on hair growth in AGA mice were firstly detected, and then the mechanisms of PAEs for AGA were predicted by integrating network pharmacology and de novo transcriptomics data of pilose antler. Finally, biological experiments were used to validate the molecular mechanism of PAEs in treating AGA both in vivo and in vitro.Results: It was found that PAEs promoted hair regrowth by accelerating the activation of anagen, delaying the anagen-catagen transition. It also alleviated the morphological changes, such as hair shortening, thinning, miniaturization, and HF number reduction, and regulated the hair regeneration process of four subtypes of hair. We further found that PAEs could promote the proliferation of HFSC, outer root sheath (ORS) cells, and hair bulb cells in AGA mice. We then integrated network pharmacology and pilose antler transcriptomics data to predict that the mechanism of PAEs treatment in AGA mice is closely related to the PI3K-AKT/Wnt-β-Catenin pathways. Subsequently, it was also verified that PAEs could activate both pathways in the skin of AGA mice. In addition, we found that PAEs perhaps increased the number of blood vessels around dermal papilla (DP) in experiments in vivo. Meanwhile, the PAEs stimulated the HFSC proliferation in vitro and activated the AKT and Wnt pathways. However, the proliferative activity of HFSC was inhibited after blocking the Wnt pathway and AKT activity.Conclusion: This study suggests that the hair growth-promoting effect of PAEs in AGA mice may be closely related to the stimulation of the AKT and Wnt pathways, which in turn activates the proliferation of HFSC.

Abstract Objective To identify the potential diagnostic biomarkers of rheumatoid arthritis (RA) and assess the relation between visuospatial dysfunction and disease activity in RA patients using mental rotation task (MRT)-based functional magnetic resonance imaging (fMRI). Methods A total of 27 RA patients (11 in remission, 16 in active) and 27 well-matched controls were enrolled. The visuospatial function of the subjects was measured by MRT. Brain activity data were collected using blood oxygen level dependent fMRI technique under MRT. Disease activity score 28 (DAS28) was used to evaluate the disease severity of RA patients. An analysis of the correlations between abnormal visuospatial-related brain regions, MRT performance, and DAS28 was conducted. Results RA patients performed worse on MRT than controls. Compared to the control group, RA patients showed enhanced activation in the left precuneus, left superior frontal gyrus and right cingulate gyrus during the rotation task, with left hemisphere dominance. RA patients in active showed enhanced activation in the left precuneus, left middle frontal gyrus and right cingulate gyrus compared to the patients in remission. The left precuneus activation was negatively correlated with MRT accuracy (r = −0.621, p = 0.01) and positively correlated with DAS28 (r = 0.710, p = 0.002), and MRT accuracy was negatively correlated with DAS28 in RA patients (r = −0.702, p = 0.002). Conclusion Enhanced activation of the left precuneus in RA patients affects visuospatial function and is closely related to disease activity. These changes may provide a valuable diagnostic neuroimaging biomarker of RA.

Objective This study explored the dynamic functional connective (DFC) alterations in patients with rheumatoid arthritis (RA) and investigated the correlation between the neuropsychiatric symptoms, peripheral inflammation and DFC alterations.Method Using resting-state functional MRI, we investigated the DFC based on spatial independent component analysis and sliding window method for 30 patients with RA and 30 healthy controls (HCs). The Spearman correlation was calculated between aberrant DFC alterations, Montreal Cognitive Assessment (MoCA), Hospital Anxiety and Depression Scale (HAD), C reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Diagnostic efficacy of indicators was assessed using receiver operating characteristic analysis (ROC).Results Three dynamic functional states were identified. Compared with HC, patients with RA showed reduced FC variabilities between sensorimotor network (SMN) and insula, SMN and orbitofrontal cortex, which were the crucial regions of sensory processing network. The above FC variabilities were correlated with the MoCA, HAD, CRP and ESR in patients with RA. Additionally, the CRP and ESR were negatively correlated to MoCA and positively related to HAD in patients with RA. The ROC analysis results showed that MoCA, HAD and FC variabilities of the sensory processing network could distinguish patients with RA from HC and also identify patients with RA with high ESR.Conclusion Our findings demonstrated that abnormal DFC patterns in sensory processing networks in patients with RA were closely associated with peripheral inflammation and neuropsychiatric symptoms. This indicates that the dynamic temporal characteristics of the brain functional network may be potential neuroimaging biomarkers for revealing the pathological mechanism of RA.

Background: Acne vulgaris is a common skin disease that affects the pilosebaceous unit, and inflammation plays an important role. Chinese traditional medicine Tanreqing (TRQ) has been used in the field of dermatology to treat inflammatory diseases and has shown outstanding efficacy. The purpose of this study was to evaluate the potential advantages of TRQ on a mouse model of acne inflammation. Additionally, network pharmacology and transcriptome analysis were utilized to investigate the possible underlying mechanisms. Methods: The active compounds of TRQ were collected through TCMSP database and literature review, and the predicted targets of active compounds were obtained from Swiss Target Prediction. Acne targets were collected from the Gene Cards database and DisGeNET. Cytoscape v3.7.1 was used to build the ''compounds-targets'' network and PPI network and to obtain the core network. GO analysis and KEGG pathway annotation were performed on the core genes. The Illumina HiSeq sequencing platform was utilized to sequence mRNA from mouse skin tissue in acne areas. Differential gene expression analysis was conducted to identify genes that were affected by TRQ, and their associated biological functions and signaling pathways were analyzed. The transcriptomic and network pharmacology analyses were integrated to generate predictive results. Additionally, the efficacy of TRQ in treating acne was confirmed through H&E staining, and the predicted outcomes were verified through quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Results: The study's results indicate that 37 potential target genes were enriched in multiple signaling pathways, including the atherosclerosis, IL-17, and TNF signaling pathways, according to network pharmacology. In addition, 338 transcriptomic DEGs were found to be involved in various signaling pathways, including Cell adhesion molecules and the IL-17 signaling pathway.The combined analysis of these findings suggests that the IL-17 and TNF signaling pathways are the primary mechanisms by which TRQ exerts its anti-acne effects. H&E staining revealed that TRQ inhibited inflammatory cell infiltration in acne lesions and attenuated the inflammatory response in acne. Moreover, TRQ significantly suppressed key factors such as TNF-α, JNK2, IL-17A, C/EBPβ, IL-6, and IL-1β in the IL-17 and TNF signaling pathways. Furthermore, TRQ also demonstrated a significant inhibition in the expression of TNF-α and IL-1β in the skin tissues of acne vulgaris mice Conclusion: Our findings suggest that TRQ may effectively inhibit the IL-17/TNF signaling pathway by suppressing the expression of TNF-α and IL-1β, making it a potential candidate for the treatment of acne

Background: Cognitive deficits are common in rheumatoid arthritis (RA) patients, but the mechanisms remain unclear. We investigated the effective connectivity and structural alterations of the core brain regions in RA patients with cognitive impairment. Methods: Twenty-four female patients with RA and twenty-four healthy controls were enrolled. We analyzed abnormal brain activity patterns using functional MRI during the Iowa gambling task (IGT) and core regions effective connectivity using dynamic causal model (DCM). Structural alterations of white matter volume (WMV) and gray matter volume (GMV) were detected using voxel-based morphometry (VBM). Results: RA patients showed altered activation patterns of the cortico-thalamo-cortical network, increased coupling strength from the left ventromedial prefrontal gyrus to the anterior cingulate cortex (ACC), the ACC to the right thalamus, and decreased connectivity from the thalamus to left hippocampus. VBM structural analysis showed increased GMV in the bilateral orbital frontal gyrus, bilateral hippocampus and right putamen, and reduced GMV and WMV in the bilateral thalamus in RA patients. Right thalamic GMV and WMV were positively correlated with the right thalamus-to-hippocampus connective strength. Additionally, the bold signal, GMV and WMV of the right thalamus were positively correlated with cognitive performance (IGT score) in RA patients. Conclusion: Results suggest a structural and functional deficiency in the cortico-thalamo-cortical network, which is characterized by increased ACC-to-thalamus strength and reduced thalamus-to-hippocampus coupling in RA patients. The cognitive dysfunction may be the result of compensatory measures against imbalanced cortico-thalamic-cortical coupling.

The coordination between regional ecological and economic development has become a crucial topic in current environmental and development research due to the establishment of sustainable development goals and the acceleration of urbanization. In this study, an improved eco-economy coordination (EEC) model is proposed to evaluate the coordination of regional ecological and economic development. This study focuses on Wuhan, China, and utilizes socioeconomic and remote sensing data from 2000 to 2015 to measure both static and dynamic ecosystem service values (ESV). ESVs are the direct and indirect benefits produced by ecosystems that support human survival and development. The calculated ESVs are then integrated into the ecological–economic coordination (EEC) evaluation. This study aims to conduct a comparative analysis of the ecological-economic coordination across various districts of Wuhan, with a focus on spatiotemporal factors. In addition, this paper highlights the role of environmental adjustment coefficients in enhancing the EEC assessment. The results show that (1) the total static ESV experienced a loss of RMB 1.2 billion (approximately USD 169 million) and showed a decreasing trend, while the dynamically corrected ESV reversed this trend; and (2) EEC based on static ESV showed a low degree of conflict between ecological and economic reconciliation compared with a significant increase in EEC based on dynamic ESV. Based on static ESV, the EEC showed a low degree of conflict between ecological and economic reconciliation compared to a significant increase in EEC based on dynamic ESV. After being corrected by environmental adjustment coefficients, the EEC values showed a more differentiated distribution. Most regions demonstrated an overall upward trend in EEC, with a few, such as Wuchang District and Jiang’an District, being notable examples. However, Jianghan District presented a high conflict status. Finally, a series of decision-making suggestions are given based on relevant study results, and an important policy implication is that the coordination of economic growth and ecological protection, under large downward pressure from EEC values, needs to be paid special attention in policy decision-making.

This study aimed to investigate the effects of the covalent binding of flaxseed protein (FP) and chlorogenic acid (CA) on the structure and functional properties of FP–CA complexes fabricated using the alkali method. The results suggested that the encapsulation efficiency of CA encapsulated by FP ranged from 66.11% to 72.20% and the loading capacity of CA increased with an increasing addition ratio of CA with a dose-dependent relationship, which increased from 2.34% to 10.19%. The particle size, turbidity, zeta potential and PDI of FP and the FP–CA complexes had no significant discrepancy. UV–Vis and fluorescence spectra showed the existence of the interaction between FP and CA. SEM images showed that the surface of the FP–0.35%CA complex had more wrinkles compared to FP. Differential scanning calorimetry analysis indicated the decomposition temperature of FP at 198 °C was higher than that (197 °C) of the FP–0.35%CA complex, implying that the stability of the FP–CA complexes was lower than FP. The functional properties suggested that the FP–CA complex with 1.40% CA had a higher water holding capacity (500.81%), lower oil holding capacity (273.495%) and lower surface hydrophobicity. Moreover, the FP–CA complexes had better antioxidant activities than that of FP. Therefore, this study provides more insights for the potential application of FP–CA covalent complexes in functional food processing.

Lactobacillus plantarum is a kind of probiotic that benefits the host by regulating the gut microbiota, but it is easily damaged when passing through the gastrointestinal tract, hindering its ability to reach the destination and reducing its utilization value. Encapsulation is a promising strategy for solving this problem. In this study, transglutaminase (TGase)-crosslinked gelatin (GE)/sodium hexametaphosphate (SHMP) hydrogels were used to encapsulate L. plantarum. The effects of TGase concentration and drying method on the physiochemical properties of the hydrogels were determined. The results showed that at a TGase concentration of 9 U/gGE, the hardness, chewiness, energy storage modulus, and apparent viscosity of the hydrogel encapsulation system were maximized. This concentration produced more high-energy isopeptide bonds, strengthening the interactions between molecules, forming a more stable three-dimensional network structure. The survival rate under the simulated gastrointestinal conditions and storage stability of L. plantarum were improved at this concentration. The thermal stability of the encapsulation system dried via microwave vacuum freeze drying (MFD) was slightly higher than that when dried via freeze drying (FD). The gel structure was more stable, and the activity of L. plantarum decreased more slowly during the storage period when dried using MFD. This research provides a theoretical basis for the development of encapsulation technology of probiotics.

ABSTRACT The rise of antibiotic resistance and dearth of novel antibiotics have posed a serious health crisis worldwide. In this study, we screened a combination of antibiotics and nonantibiotics providing a viable strategy to solve this issue by broadening the antimicrobial spectrum. We found that chenodeoxycholic acid (CDCA), a cholic acid derivative of the traditional Chinese medicine (TCM) Tanreqing (TRQ), synergizes with amikacin against Staphylococcus aureus in vitro, and this synergistic killing was effective against diverse methicillin-resistant S. aureus (MRSA) variants, including small-colony variants (SCVs), biofilm strains, and persisters. The CDCA-amikacin combination protects a mouse model from S. aureus infections. Mechanistically, CDCA increases the uptake of aminoglycosides in a proton motive force-dependent manner by dissipating the chemical potential and potentiates reactive oxygen species (ROS) generation by inhibiting superoxide dismutase activity. This work highlights the potential use of TCM components in treating S. aureus-associated infections and extend the use of aminoglycosides in eradicating Gram-positive pathogens. IMPORTANCE Multidrug resistance (MDR) is spreading globally with increasing speed. The search for new antibiotics is one of the key strategies in the fight against MDR. Antibiotic resistance breakers that may or may not have direct antibacterial action and can either be coadministered or conjugated with other antibiotics are being studied. To better expand the antibacterial spectrum of certain antibiotics, we identified one component from a traditional Chinese medicine, Tanreqing (TRQ), that increased the activity of aminoglycosides. We found that this so-called agent, chenodeoxycholic acid (CDCA), sensitizes Staphylococcus aureus to aminoglycoside killing and protects a mouse model from S. aureus infections. CDCA increases the uptake of aminoglycosides in a proton motive force-dependent manner by dissipating the chemical potential and potentiates ROS generation by inhibiting superoxide dismutase activity in S. aureus. Our work highlights the potential use of TCM or its effective components, such as CDCA, in treating antibiotic resistance-associated infections.

Prevalent methods for monitoring burn injuries and testing drug efficacies rely on fixed tissue sections. However, this may leave out cellular details in the living state. In vivo assessments of burn healing has been long sought after and are of scientific and clinical interest. Nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), collagen, and melanin are endogenous fluorescent molecules and their signals can be captured by two-photon microscopy (TPM), therefore providing information on epidermal histological features and collagen growth in real-time. In addition, TMP imaging on exogenous fluorescent substances provides a basis for detecting blood vessels. In this work, two-photon microscopy was used to capture the exogenous fluorescent substances and endogenous fluorescent molecules at different times to assess and track burn healing in vivo. Combining TPM imaging and morphological characteristics, proliferation and differentiation of the keratinocytes in different layers of skin, collagen contents, and angiogenesis were identified and quantified. The TPM monitoring method provides an effective tool to systemically evaluate skin healing of deep burns in vivo.

AbstractBackground Abdominal wound dehiscence (AWD) is a major complication of abdominal surgery, and neonates are a group with a high risk of AWD, which has serious consequences or can even result in death. The purpose of this study is to explore the risk factors for neonatal AWD and construct a predictive model.Methods The clinical data of 453 cases that underwent neonatal laparotomy from June 2009 to June 2020 were retrospectively analyzed, among which 27 cases of AWD were identified. Nine factors, including gender, age at admission, weight at admission, preterm delivery, level of preoperative anaemia, hypoalbuminemia, operation time, incision length, and incision type, were analyzed to explore their correlation with neonatal AWD.Results The incidence of neonatal AWD was 6.0% (27/453), among which partial wound dehiscence accounted for 4.9% (22/453) and complete wound dehiscence accounted for 1.1% (5/453). Hypoproteinemia and incision type were the independent risk factors for neonatal AWD, and weight at admission was a protective factor for AWD in the multivariate models. All these factors were incorporated to construct a nomogram, and a calibration curve was plotted. The result indicated that the actual risk was close to the predicted risk when the predicted risk rate was greater than about 35%.Conclusions Neonatal AWD is closely related to hypoproteinemia and incision contamination. Our predictive model showed the potential to provide an individualized risk estimate of AWD for neonatal patients undergoing abdominal surgery.Key messagesNeonatal abdominal wound dehiscence (AWD) has a serious consequence and the incidence of neonatal AWD was about 6.0% and the complete AWD morbidity is 1.1%.Hypoproteinemia and incision type were the independent risk factors for neonatal AWD.Our predictive model showed the potential to provide an individualized risk estimate of AWD for neonatal patients undergoing abdominal surgery.

The transdermal administration of collagen is an important method used for wound healing and skin regeneration. However, due to the limitations of previous approaches, the process and degree of collagen transdermal absorption could only be quantitatively and qualitatively evaluated in vitro. In the present study, we introduced a novel approach that combines second-harmonic generation with two-photon excited fluorescence to visualize the dynamics of collagen transdermal absorption in vivo. High-resolution images showed that exogenous recombinant human collagen permeated the epidermis through hair follicles and sebaceous glands reached the dermis, and formed reticular structures in real time. We also validated these findings through traditional in vitro skin scanning and histological examination. Thus, our approach provides a reliable measurement for real-time evaluation of collagen absorption and treatment effects in vivo.

The primary goal of wound healing therapy is to quickly close the wound and form healthy and functional scar tissue. As capillary system regeneration is an important condition for skin wound healing, it is important to dynamically monitor angiogenesis with multiple methods based on the 3D reconstruction of dermal microcirculation. In this study, we established a quantitative system to comprehensively evaluate angiogenesis and perfusion during wound healing. Full-thickness skin wounds were generated on the back of rats. Laser Doppler anemometer (LDA), laser speckle contrast imaging (LSCI), and medical electronic dermoscopy (MED) are used to observe the vasculature from the horizontal plane and quantify blood perfusion in wounds. We combined immunofluorescence staining and two-photon excited fluorescence microscopy (TPEF) to reconstruct 3D vessels for longitudinal quantification. These results indicated that new blood vessels around the wound grew parallel to the wound surface. The shape of the blood vessels changed from curved to straight, and the density of the surrounding vascular network gradually decreased as manifested by a decrease in the nodes and branches and the total length of the blood vessels. There were dense blood vessels that grew vertically to the wound surface in the center of the wound. This study established a real-time, non-invasive system to monitor re-epithelialization and angiogenesis and quantitatively assess blood vessels during wound healing, which facilitates the evaluation of new treatments for wound healing.

Staphylococcus aureus has been recognized as an important human pathogen and poses a serious health threat worldwide. With the advent of antibiotic resistance, such as the increased number of methicillin-resistant Staphylococcus aureus (MRSA), there is an urgent need to develop new therapeutical agents. In this study, Chinese traditional medicine Tanreqing (TRQ) has been used as an alternative treating agent against MRSA and we aim to unravel the mode of action of TRQ underlying MRSA inhibition. TRQ treatment affected numerous gene expression as revealed by RNA-seq analysis. Meanwhile, TRQ targeted cell division to inhibit cell growth as shown by illumination microscopy. Besides, we confirmed that TRQ downregulates the expression of virulence factors such as hemolysin and autolysin. Finally, we used a murine model to demonstrate that TRQ efficiently reduces bacterial virulence. Altogether, we have proved TRQ formula to be an effective agent against S. aureus infections.

Objective: This study aimed to explore the structural changes in patients with subclinical hypothyroidism (SCH) using voxel-based morphometry (VBM) and to investigate the altered attentional control networks using functional MRI (fMRI) during the performance of a modified Stroop task with Chinese characters.Methods: High-resolution three-dimensional (3D) T1-weighted images and an fMRI scan were taken from 18 patients with SCH and 18 matched control subjects. The Montreal Cognitive Assessment Chinese-revised (MoCA-CR) and the Stroop task were used to evaluate the cognitive and attention control of the participants.Results: Compared to controls, the VBM results showed decreased gray matter volumes (GMVs) in bilateral prefrontal cortices (PFCs, including middle, medial, and inferior frontal gyri), cingulate gyrus, precuneus, left middle temporal gyrus, and insula in patients with SCH. The fMRI results showed a distributed network of brain regions in both groups, consisting of PFCs (including superior and middle and inferior frontal cortices), anterior cingulate cortex (ACC), posterior cingulate cortex, and precuneus, as well as the insula and caudate nucleus. Compared to controls, the SCH group had lower activation of the above brain areas, especially during the color-naming task. In addition, the normalized GMV (nGMV) was negatively correlated with thyroid-stimulating hormone (TSH) level (r = −0.722, p < 0.001).Conclusion: Results indicate that patients with SCH exhibit reduced GMVs, altered BOLD signals, and activation in regions associated with attention control, which further suggest that patients with SCH may have attentional control deficiency, and the weakened PFC–ACC–precuneus brain network might be one of the neural mechanisms. Negative correlations between nGMV and TSH suggest that TSH elevation may induce abnormalities in the cortex.

Methicillin-resistant Staphylococcus aureus (MRSA) has posed a severe global health threat. In this study, we screened an antibiotic and non-antibiotic combination that provides a viable strategy to solve this issue by broadening the antimicrobial spectrum. We found that chenodeoxycholic acid (CDCA) could synergistically act with carbapenem antibiotics to eradicate MRSA-related infections. This synergy specifically targets MRSA and was also validated using 25 clinical MRSA strains using time–kill analysis. We speculated that the underlying mechanism was associated with the interaction of penicillin-binding proteins (PBPs). As a result, the synergistic efficiency of CDCA with carbapenems targeting PBP1 was better than that of β-lactams targeting PBPs. Moreover, we showed that CDCA did not affect the expression level of PBPs, but sensitized MRSA to carbapenems by disrupting the cell membrane. In our study, we have revealed a novel synergistic combination of antibiotics and non-antibiotics to combat potential bacterial infections.

This paper focuses on improving thermal efficiency and reducing unburned carbon in fly ash by optimizing operating parameters via a novel high-efficient swarm intelligence optimization algorithm (grey wolf optimizer algorithm, GWO) for coal-fired boiler. Mathematical models for thermal efficiency and unburned carbon in fly ash of the discussed boiler are established by artificial neural network (ANN). Based on the ANN models, the grey wolf optimizer algorithm is used to obtain higher thermal efficiency and lower unburned carbon by optimizing the operating parameters. Meanwhile, the comparisons between GWO and particle swarm optimization (PSO) and genetic algorithm (GA) show that GWO has superior performance to GA and PSO regarding the boiler combustion optimization. The proposed method can accurately optimize the boiler combustion performance, and its validity and feasibility have been experimentally validated. Additionally, a run of optimization takes a less time period, which is suitable for the real-time optimization.

In view of the problems of long matching time and the high-dimension and high-matching rate errors of traditional scale-invariant feature transformation (SIFT) feature descriptors, this paper proposes an improved SIFT algorithm with an added stability factor for image feature matching. First of all, the stability factor was increased during construction of the scale space to eliminate matching points of unstable points, speed up image processing and reduce the dimension and the amount of calculation. Finally, the algorithm was experimentally verified and showed excellent results in experiments on two data sets. Compared to other algorithms, the results showed that the algorithm proposed in this paper improved SIFT algorithm efficiency, shortened image-processing time, and reduced algorithm error.

Abstract Background Combining conventional drugs and traditional medicine may represent a useful approach to combating antibiotic resistance, which has become a serious threat to global public health. This study aimed to evaluate the potential synergistic interactions between Tanreqing (TRQ) injection, a commercial traditional Chinese medicine formula used for the treatment of upper respiratory tract infection, and selected antibiotics used against methicillin-resistant Staphylococcus aureus (MRSA). Methods The minimum inhibitory concentrations (MICs) of TRQ, vancomycin and linezolid against planktonic MRSA strain were determined by the broth microdilution method. The combined effects of TRQ and antibiotics were studied by the checkerboard method and the time-kill curve assay. The 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay was employed to determine the inhibitory effect of the test compounds alone and in combination against MRSA embedded in biofilms. Results MRSA strain was found to be susceptible to TRQ formula with MIC value 4125 μg/ml, while the MIC values for antibiotics, vancomycin and linezolid, were 2.5 μg/ml. The checkerboard analysis revealed that TRQ markedly enhanced activities of the tested antibiotics by reducing their MICs. In the time-kill analysis, TRQ at 1/2 × MIC in combination with vancomycin at 1/2 × MIC, as well as TRQ at 1/8 × MIC in combination with linezolid at 1/2 × MIC decreased the viable colonies by ≥2log10 CFU/ml, resulting in a potent synergistic effect against planktonic MRSA. In contrast to the tested antibiotics, which did not affect mature MRSA biofilms at subinhibitory concentrations, TRQ alone showed strong ability to disrupt preformed biofilms and induce biofilm cell death. The combination of TRQ with vancomycin or linezolid at sub-MIC concentrations resulted in a synergistic antibiofilm effect significantly higher than for each single agent. Conclusions This study provides the first in vitro evidence on the synergistic effects of TRQ and vancomycin or linezolid against planktonic and biofilm MRSA, and revealed their optimal combination doses, thereby providing a rational basis for the combination therapies against MRSA.

Abstract In this study, a novel 27-locus system (now known as the SureID PanGlobal system), including 24 autosomal STRs (D3S1358, TH01, D21S11, D18S51, Penta E, D12S391, D6S1043, D2S1338, D1S1656, D2S441, D5S818, D13S317, D7S820, D19S433, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D16S539, D22S1045, SE33, D10S1248), two Y-chromosome markers (DYS391 and Y-indel) and the sex determining marker, Amelogenin was developed with six fluorescent dyes labeling. The included STR loci belonged to the core loci in the Combined DNA Index System (CODIS) and the European Standard Set (ESS) as well as some additional loci commonly used in commercial kits and national DNA databases. This paper describes the validation studies conducted with the SureID PanGlobal system using Applied Biosystems 3500 XL Genetic Analyzer for fragment length detection that included the analysis of the following parameters and aspects: PCR conditions, sensitivity, species specificity, inhibition, precision, stutter, DNA mixtures, and stability studies with crime scene samples. The studies demonstrated that the SureID PanGlobal system is reproducible, accurate, sensitive and robust for forensic application and databasing. Additionally, the whole cycling time of the system can finish within 65 minutes, which was developed specifically for rapid and reliable generation of DNA profiles obtained from blood, buccal swabs and forensic stains.

Due to its accuracy, sensitivity and high throughput, real time quantitative PCR (RT-qPCR) has been widely used in analysing gene expression. The quality of data from such analyses is affected by the quality of reference genes used. Expression stabilities for nine candidate reference genes widely used in soybean were evaluated under different stresses in this study. Our results showed that EF1A and ACT11 were the best under salinity stress, TUB4, TUA5 and EF1A were the best under drought stress, ACT11 and UKN2 were the best under dark treatment, and EF1B and UKN2 were the best under virus infection. EF1B and UKN2 were the top two genes which can be reliably used in all of the stress conditions assessed.