LINE-1 transcription activates long-range gene expression.

Long interspersed nuclear element-1 (LINE-1 or L1) is a retrotransposon group that constitutes 17% of the human genome and shows variable expression across cell types. However, the control of L1 expression and its function in gene regulation are incompletely understood. Here we show that L1 transcription activates long-range gene expression. Genome-wide CRISPR-Cas9 screening using a reporter driven by the L1 5' UTR in human cells identifies functionally diverse genes affecting L1 expression. Unexpectedly, altering L1 expression by knockout of regulatory genes impacts distant gene expression. L1s can physically contact their distal target genes, with these interactions becoming stronger upon L1 activation and weaker when L1 is silenced. Remarkably, L1s contact and activate genes essential for zygotic genome activation (ZGA), and L1 knockdown impairs ZGA, leading to developmental arrest in mouse embryos. These results characterize the regulation and function of L1 in long-range gene activation and reveal its importance in mammalian ZGA.

Porous NiMoO4 Nanosheet Films and a Device with Ultralarge Optical Modulation for Electrochromic Energy-Storage Applications.

Reducing building energy consumption, improving aesthetics, and improving occupant privacy as well as comfort by dynamically adjusting solar radiation are important application areas for electrochromic (EC) smart windows. However, the current transition metal oxides still cannot meet the requirements of neutral coloration and large optical modulation. We report NiMoO4 nanosheet films directly grown on fluorine-doped tin oxide glasses. The as-grown NiMoO4 film not only achieves neutral coloration from transparent to dark brown but also shows an ultralarge optical modulation (86.8% at 480 nm) and excellent cycling stability (99.4% retention of maximum optical modulation after 1500 cycles). Meanwhile, an EC device demonstrating good EC performance was constructed. These results will greatly promote the research and development of binary transition metal oxides for both EC and energy-storage applications, and NiMoO4 films may be an excellent candidate to replace NiO films as ion-storage layers in complementary EC devices with WO3 films as EC layers.

Vegetation restoration restrains rill erosion on slag heaps in high-altitude goldfields.

Soil erosion leads to soil degradation and depletion of land resources, posing a significant threat to industrial production and ecological sustainability. In high-altitude regions, rill erosion is the main form of soil erosion in mining areas, however, our understanding of morphology and developmental characteristics of rills and the mechanisms influencing them remains limited. In this study, data were collected from 96 rill plots across two gold mines in the eastern Tibetan Plateau according to vegetation restoration modes (natural restoration (CK) and planted with Elymus dahuricus (ED), Medicago sativa (MS), and multi-plant mixed (Avena fatua L. + Elymus dahuricus + Medicago sativa + Oxytropis coerulea, MM)) and restoration periods (1 year, 3 years, 4 years, and 6 years). We investigated the variations of 7 indicators that can reveal rill morphological and developmental characteristics across different restoration modes and restoration periods, and utilized a partial least squares structural equation model (PLS-SEM) to analyze the effects of 15 indicators from topography, soil, and vegetation on rill erosion modulus (REM). The results indicated that artificial vegetation restoration effectively restrained rill development, notably by decreasing the frequency of wider (>15 cm) and deeper (>10 cm) rills when compared to CK plots. Planting MM and ED exhibited greater efficacy in controlling rill erosion than planting MS. However, the effectiveness of planting ED in controlling rill erosion gradually weakened with time, while MM consistently maintained a strong inhibitory effect. Topographic features, soil texture, and vegetation significantly influenced the REM through direct or indirect effects. Plant root functional traits were the main driving factors in reducing REM, affecting not only REM directly but also influencing vegetation-induced soil properties to indirectly effect REM.

Multi-longitudinal mode laser beat-frequency optical fiber vibration sensing system based on an FM radio module.

A multi-longitudinal mode (MLM) laser beat-frequency optical fiber vibration sensor using a frequency modulation (FM) radio integrated circuit module as the FM demodulation scheme is presented and demonstrated. To the best of our knowledge, this is the first case where a fiber-optic sensing system is combined with an FM radio module, and dynamic sensing is well achieved. As the carrier of the vibration source, the beat-frequency signal (BFS) generated by the MLM laser is transmitted to the FM radio module for FM and demodulation. The experimental results show that the system can successfully detect the vibration signal in the frequency range of 20 Hz to 18 kHz and accurately demodulate the waveform and amplitude of the vibration signal source. The minimum shape variable detected by the system is 20.67 nm, based on the performance of the commercial FM radio module itself, which can effectively solve the problem of detecting tiny vibration. The idea of the optical fiber vibration sensing system is extremely innovative, with high sensitivity, high signal-to-noise ratio (SNR), good stability, and strong resistance to electromagnetic interference.

Topography shapes the carbon allocation patterns of alpine forests.

Topography plays a crucial role in determining the structure of alpine forests, as it restricts the availability of nutrients and water necessary for plant growth. Nevertheless, our information on how variations in forest carbon allocation patterns driven by fine-scale topography are influenced by broader-scale environmental contexts is limited. In the northern Tibetan Plateau, we combined field data from 89 forest plots with a high-resolution (1 m2) digital elevation model (DEM) and utilized a linear mixed-effects model to investigate how microtopography (characterized by slope, aspect, and topographic wetness index (TWI)) and broader-scale environmental context (characterized by elevation) and their interactions affect the carbon allocation patterns of alpine forest. Our results revealed that at low and high elevations with pronounced subsurface resource limitations, plants tend to allocate a higher proportion of carbon to the root system and have lower aboveground carbon stocks (ACS). Microtopographic heterogeneity significantly influenced the carbon allocation patterns of forest, with the intensity and direction of these effects varying across the environmental gradient. At low elevations, topographically wetter and northerly microhabitats had higher ACS and lower ratios of below- and aboveground carbon stocks (RBA); however, at high elevations, topographically drier and southerly microhabitats had higher ACS and lower RBA. TWI and aspect had the weakest effect on ACS and RBA in the mid-elevations. The relationship between slope and ACS and RBA was significantly positive but not evidently related to the broader-scale environmental gradient.

Constructing a Tregs-associated signature to predict the prognosis of colorectal cancer patients: A STROBE-compliant retrospective study.

Colorectal cancer (CRC) ranks as the second leading cause of cancer-related mortality worldwide. Regulatory T cells (Tregs) are a key constituent of immune cells in the tumor microenvironment (TME) and are significantly associated with patient outcomes. Our study aimed to construct a Treg-associated signature to predict the prognosis of CRC patients. The genes' expression values and patients' clinicopathological features were downloaded from TCGA and gene expression omnibus (GEO) databases. The single-cell RNA (scRNA) sequencing data of CRC were analyzed through the Deeply Integrated human Single-Cell Omics database. WGCNA analysis was used to select Tregs-associated genes (TrAGs). The infiltrated levels of immune and stromal cells were accessed through the ESTIMATE algorithm. Cox regression analysis and the LASSO algorithm were implemented to construct prognostic models. Gene set enrichment analysis (GSEA) was performed to annotate enriched gene sets. Based on scRNA sequencing data, our study uncovered that more Tregs were significantly enriched in the TME of CRC. Then we identified 123 differentially expressed TrAGs which mainly participated in immune regulation. Given that CRC patients were reclassified into 2 subgroups with distinct overall survival based on 26 differentially expressed TrAGs with prognostic values, we subsequently constructed a signature for CRC. After training and validating in independent cohorts, we proved that this prognostic model can be well applied to predict the prognosis of CRC patients. Further analysis exhibited that more tumor-suppressing immune cells and higher immune checkpoint genes were enriched in CRC patients with high-risk scores. Moreover, immunohistochemistry analysis validated that the genes in the prognostic model were significantly elevated in CRC tissues. We were the first to construct a prognostic signature for CRC based on TrAGs and further revealed that the poor prognosis of patients was mainly attributed to the tumor-suppressing microenvironment and upregulated immune checkpoint genes in tumor tissues.

Resina Draconis Particles Encapsulated in a Hyaluronic-Acid-Based Hydrogel to Treat Complex Burn Wounds.

Severe burns require urgent new dressing treatments due to their irregular wounds and secondary injuries associated with dressing changes. In this study, a hyaluronic-acid-based hydrogel was developed to treat complex burn wounds. This hydrogel was prepared by mixing and cross-linking oxidized hyaluronic acid (OHA) and carboxymethyl chitosan (CMCS) through Schiff base reactions. Micronized Resina Draconis particles were encapsulated in this hydrogel to achieve sustained release of the active components when applied on wounds. The Resina-Draconis-loaded hydrogel (RD-Gel) demonstrated good mechanical properties and excellent self-healing. The results of in vitro experiments confirmed that RD-Gel had good biocompatibility, and was able to enhance cell migration and inhibit the production of inflammatory cytokines. It also induced rapid hemostasis in rats, downregulated the levels of inflammatory cytokines, and promoted collagen regeneration on model animals, eventually accelerating the rebuilding of skin structures and wound recovery.

A self-healing hydrogel based on crosslinked hyaluronic acid and chitosan to facilitate diabetic wound healing.

Traditional wound dressings are not able to provide ideal environment for diabetic wounds surface thus hampered the regrowth of fresh tissues. In this study, we designed a novel in situ forming hydrogel and used it as wound dressing material. Carboxymethyl chitosan (CMCS) and oxidized hyaluronic acid (OHA) were selected to construct a pH-responsive and self-healing hydrogel system via Schiff base reaction. Taurine (Tau) with anti-inflammatory property was loaded in the hydrogel through the aforementioned reaction. Under the slightly acidic environment of the diabetic wound site, a responsive release of taurine molecules speeded up the transfer of the taurine into the wound. The physiochemical properties of the prepared CMCS-OHA-Tau hydrogel were characterized. The CMCS-OHA-Tau hydrogel showed good biocompatibility, enhancement of cell migration and inhibited production of inflammatory cytokines.Subsequently, the hydrogel was applied on the wounds of diabetic rats and its boosted efficacy for wound recovery was confirmed.

Aldehyde spiropyran fluorescent probe for rapid determination of hydrazine in environmental water.

Hydrazine often receives publicity because it has a wide range of applications but high toxicity at the same time. Here, we invented a novel aldehyde spiropyran fluorescent probe (SP-CHO) for fluorescence determination of hydrazine. The probe was synthesized through a hydrolysis reaction and a condensation reaction. It exhibits a specific response to hydrazine. The influence factors and anti-interference ability of SP-CHO identifying hydrazine were studied. HRMS, 1 H NMR, and density functional theory (DFT) calculations were used to reveal the recognition mechanism. Results showed that SP-CHO can be used for fluorescence determination of hydrazine with high selectivity and sensitivity. An SP-CHO-based fluorescence method was established for quantitation of hydrazine. The detection limit was 1.26 µmol/l, and the linear range was 5-100 µmol/l. The determination of hydrazine in water samples can be completed within 10 min, which shows good application prospects in real-time detection and process monitoring.

Oxidation-Responsive Micelles for Drug Release Monitoring and Bioimaging of Inflammation Based on FRET Effect in vitro and in vivo.

Purpose: A new approach to monitor drug release and image inflammatory reactions in vitro and in vivo based on FRET mechanism was reported. Methods: In this study, mixed micelles containing a synthesized fluorescent donor DAN-PPS-mPEG and its quencher DAB-PPS-mPEG were prepared. Their stabilities, self-assembling and oxidation-responsiveness towards oxidants were tested in vitro and in vivo. Results: The conjugated polymers were synthesized and the morphological change and the fluorescent spectra of the prepared micellar system were measured. After incubating the DAN/DAB-PPS-mPEG mixed micelles with stimulated L929 fibroblast cells, the result of confocal laser microscopy showed fluorescence restoration of the micelles. Furthermore, an acute inflammatory injury mouse model was used to test the micelles in vivo. The micelles showed its ability to visualize the inflammatory site in the abdomen of the mice. Conclusion: The results confirmed that DAN/DAB-PPS-mPEG mixed micelles can respond to oxidants and release encapsulated cargos with corresponding fluorescence restoration, and visualize the inflammatory cells in vitro and inflammatory reactions in vivo.

Fabrication of highly sensitive and uniform Ag/PS/PDMS SERS substrate and its application forin-situdetection.

In this study, we developed a flexible and transparent silver/polystyrene/polydimethylsiloxane (Ag/PS/PDMS) substrate with both high density of hot spots and satisfactory uniformity using a cost-effective approach. Via template-guided self-assembly, PS beads were arranged regularly in nanobowls of a square array on PDMS, whose surface structure was transferred from a commercial complementary metal oxide semiconductor chip. Roughness was introduced onto the PS bead surface by nitrogen plasma treatment, followed by sputtering of Ag which generated many hot spots. Differential roughness on the PS bead surface greatly influenced the morphology of the Ag/PS/PDMS substrate. A meat-ball like surface structure was formed with a plasma etching time of 5 min, whose growth mechanism was proposed based on the scanning electron microscope analysis. The high sensitivity and desirable uniformity of the meat-ball like Ag/PS/PDMS substrate were demonstrated by using crystal violet as a Raman reporter, exhibiting an enhancement factor of 2.7 × 107and a relative standard deviation of 5.04%. Thiram of a lower concentration than the maximum residue limit on the cucumber surface could easily be detectedin situby the proposed substrate, demonstrating its great potential forin-situfood safety analysis.

Improving Water-Absorption and Mechanical Strength: Lyotropic Liquid Crystalline-Based Spray Dressings as a Candidate Wound Management System.

A spray dressing based on lyotropic liquid crystalline (LLC) with adjustable crystalline lattices was investigated in this study. It possesses water-triggering phase transition property and ease of spraying on wound, as well as stable drug encapsulation and controllable drug release. When it comes to wound with exudate, adequate water absorption and sustainable mechanical strength after water absorption was important for a good dressing, while most of the normal LLC dressings were still unable to meet such standards. Herein, a type of hyaluronic acid (HA)-incorporated LLC-based spray dressing (HLCSD) was developed to overcome the above limitations. After comparing HAs with different molecular weights (MWs) and concentrations, 3% HA with MW of 800~1000 kD was chosen as an ideal amount of excipients to add into the HLCSD. The water absorption of HLCSD precursor increased by 150% with the appearance of enlarged water channels. The complex modulus of HLCSD gel also increased from 1 to 100 kPa, which suggested lasting wound coverage and good patient compliance when used clinically. The spraying and phase transition properties of HLCSD was studied and showed acceptable changes. Moreover, good safety comparable with the commercial product Purilon® was also demonstrated in an in vivo acute skin irritation test. Thus, the improved HLCSD was a promising dressing for exudation wound treatment.

Singular Configuration Analysis and Singularity Avoidance with Application in an Intelligent Robotic Manipulator.

Recently, robotic sensor systems have gained more attention annually in complex system sense strategies. The robotic sensors sense the information from itself and the environment, and fuse information for the use of perception, decision, planning, and control. As an important supplement to traditional industrial robots, co-bots (short for co-working robots) play an increasingly vital role in helping small and medium-sized enterprises realize intelligent manufacturing. They have high flexibility and safety so that they can assist humans to complete highly repetitive and high-precision work. In order to maintain robot safe operation in the increasing complex working environment and human-computer intelligent interactive control, this paper is concerned with the problem of applicant accuracy analysis and singularity avoidance for co-bots. Based on the dynamic model with load and torque sensors, which is used to detect the external force at the end of the robot, this paper systematically analyzes the causes of singularity phenomenon in the robot motion control. The inverse solution is obtained by analytical method and numerical method, respectively. In order to ensure the smooth and safe operation in the whole workspace, it is necessary for a robot to avoid singularity. Singularity avoidance schemes are utilized for different control tasks, including point-to-point control and continuous path control. Corresponding simulation experiments are designed to verify the effectiveness of different evasion schemes, in which the advantages and disadvantages are compared and analyzed.

A bacteria-resistant and self-healing spray dressing based on lyotropic liquid crystals to treat infected post-operative wounds.

The delayed healing of infected post-operative wounds has turned into a worldwide medical problem. In the clinical treatment, effective bacterial clearance and promoted wound healing were considered as two crucial aspects. However, the effect of current dressings with antibacterial activity was limited due to the declined efficacy against antibiotic-resistant bacteria, and poor mechanical property during skin extension and compression movement. In this project, a lyotropic liquid crystal (LLC)-based bacteria-resistant and self-healing spray dressing loaded with ε-polylysine (PLL) was designed. Owing to the unique antibacterial mechanism, PLL was expected to kill antibiotic-resistant bacteria efficiently, even the "superbug" methicillin-resistant Staphylococcus aureus (MRSA). The cubic cells of LLC were applied to encapsulate PLL to improve its stability and induce a sustained release, further realizing a long-term antibacterial effect. Meanwhile, the LLC precursor (LLCP) could extend to the irregular edges of the wound, and spontaneously transited to a cubic phase gel once exposed to physiological fluid. This 3D structure was also endowed with mechanically responsive viscoelasticity that formed a robust and flexible defense for wounds. An excellent antibacterial activity with more than 99% MRSA killed in 3 h was demonstrated by a killing kinetics study. The long-term effect was also proved by measuring the bacteriostatic circle test within 48 h. In addition, the unique sol-gel phase transition behavior and superior self-healing capacity of PLL-LLCP was verified with the rheological study and self-recoverable conformal deformation test in vivo. In the infected post-operative wound model, satisfactory bacterial clearance and prominent wound healing promotion were realized by PLL-LLCP, with the survival of the bacteria at lower than 0.1% and the wound closure at higher than 90%. Thus, PLL-LLCP was believed to be an excellent candidate for the therapy of infected post-operative wounds.

Myeloid-derived suppressor cells infiltration in non-small-cell lung cancer tumor and MAGE-A4 and NY-ESO-1 expression.

Cancer/testis antigens melanoma-associated antigen 4 (MAGE-A4) and New York esophageal squamous cell carcinoma-1 (NY-ESO-1) are of clinical interest as biomarkers and present valuable targets for immunotherapy; however, they are poor prognostic markers in non-small cell lung cancer (NSCLC). In addition, myeloid derived suppressor cells (MDSCs) are recognized as a key element in tumor escape and progression. The aim of the present study was to investigate the diagnostic and prognostic value of MAGE-A4 and NY-ESO-1, and their association with MDSCs in NSCLC samples. The expression levels of MAGE-A4 and NY-ESO-1, and the infiltration of MDSCs (CD33+), were analyzed by immunohistochemistry of 67 tissue samples from patients with NSCLC. Overall, 58.33% of the NSCLC squamous cell carcinoma tissues and 94.7% of adenocarcinoma tissues were positive for MAGE-A4. NY-ESO-1 expression was observed in 52.78% of the squamous cell carcinoma tissues and 80% of the adenocarcinoma tissues. In primary adenocarcinoma tumor tissues, MAGE-A4 and NY-ESO-1 demonstrated a higher intensity of expression compared with the squamous cell carcinoma tissues. A total of 33 (91.7%) squamous cell carcinoma and 19 (95.0%) adenocarcinoma specimens were positive for CD33. The expression of MAGE-A4 and NY-ESO-1 antigens and infiltration of MDSCs was associated with poor prognosis of patients with NSCLC. Further studies investigating the association between these findings and underlying molecular mechanisms are required.

MicroRNA-221 promotes breast cancer resistance to adriamycin via modulation of PTEN/Akt/mTOR signaling.

As a prevalent tumor among women, breast cancer is still an incurable disease due to drug resistance. In this study, we report microRNA-221 to have a significant effect on breast cancer resistance to adriamycin. The microRNA-221 is elevated in tumor tissue compared with nearby nontumor samples, as well as in breast cancer cell line with adriamycin resistance (MCF-7/ADR) compared to its parental line (MCF-7) and the normal breast epithelial cell line (MCF-10A). Enforced level of microRNA-221 promotes cancer resistance to adriamycin, which in turn sustains cell survival and exacerbates malignant formation. Reciprocally, the silence of microRNA-221 in cancer cells augments the sensitivity to chemotherapy, thereby resulting in enhanced apoptosis of MCF-7/ADR cells. Mechanistically, we identify PTEN as a direct target of microRNA-221, which was conversely associated with a microRNA-221 level in breast tumors. The knock-down of PTEN partially reversed the stimulatory role of microRNA-221 in the modulation of the Akt/mTOR signaling. Taken together, these findings suggest microRNA-221 suppresses PTEN transcription and activates Akt/mTOR pathway, which in turn enhances breast cancer resistance to adriamycin and promotes cancer development. Our data thus illuminate the microRNA-221/PTEN axis may act as a promising strategy for the treatment of chemotherapy-resistant breast tumors.

Polyaniline Nanofiber Wrapped Fabric for High Performance Flexible Pressure Sensors.

The rational design of high-performance flexible pressure sensors with both high sensitivity and wide linear range attracts great attention because of their potential applications in wearable electronics and human-machine interfaces. Here, polyaniline nanofiber wrapped nonwoven fabric was used as the active material to construct high performance, flexible, all fabric pressure sensors with a bottom interdigitated textile electrode. Due to the unique hierarchical structures, large surface roughness of the polyaniline coated fabric and high conductivity of the interdigitated textile electrodes, the obtained pressure sensor shows superior performance, including ultrahigh sensitivity of 46.48 kPa-1 in a wide linear range (<4.5 kPa), rapid response/relaxation time (7/16 ms) and low detection limit (0.46 Pa). Based on these merits, the practical applications in monitoring human physiological signals and detecting spatial distribution of subtle pressure are demonstrated, showing its potential for health monitoring as wearable electronics.

[Mid-term effectiveness of Coflex interspinous dynamic internal fixation combined with spinal fusion for lumbar disc degeneration].

Objective: To evaluate the effectiveness of Coflex interspinous dynamic internal fixation combined with spinal fusion for lumbar disc degeneration. Methods: The clinical data of 39 patients with two-level lumbar disc degeneration who met the selection criteria between June 2010 and December 2011 was retrospectively analyzed. They were divided into group A (20 cases, simple lumbar decompression and fusion) and group B (19 cases, Coflex interspinous dynamic internal fixation combined with spinal fusion) according to different surgical methods. There was no significant difference in age, gender, disease diagnosis, lesion segment, disease duration, Oswestry disability index (ODI), visual analogue scale (VAS) score, and the intervertebral height, foramen intervertebral height (FIH), and range of motion (ROM) of upper operative segment and adjacent segment between the two groups ( P>0.05). ODI and VAS score were used to evaluate the effectiveness before operation and at last follow-up, and the improvement rates were calculated. The intervertebral height [anterior disc height (ADH), middle disc height (MDH), and posterior disc height (PDH)], FIH, and ROM were measured and compared between the two groups. Results: The operation time and intraoperative blood loss in group A were significantly more than those in group B ( P<0.05), and there was no significant difference in hospitalization time between the two groups ( t=0.992, P=0.328). All patients were followed up; the follow-up time was 33-50 months (mean, 40.5 months) in group A and 39-51 months (mean, 42.6 months) in group B. No complication such as displacement, loosening, or rupture of internal fixator was found in both groups. At last follow-up, ODI and VAS score of the two groups significantly improved when compared with preoperative scores ( P<0.05). At last follow-up, there was no significant difference in ODI, VAS score, and improvement rate of ODI between the two groups ( P>0.05); the improvement rate of VAS score in group B was significantly higher than that in group A ( t=2.245, P=0.031). There was no significant difference in the intervertebral height and FIH of the upper operative segment at last follow-up between the two groups and between preoperation and last follow-up in the two groups ( P>0.05). At last follow-up, the ADH of adjacent segment in group B was significantly higher than that in group A, and MDH, PDH, and FIH were significantly lower than those in group A ( P<0.05). Compared with preoperation, the ADH of adjacent segment in group A decreased and MDH, PDH, and FIH increased at last follow-up ( P<0.05), while all indexes in group B did not change significantly ( P>0.05). The ROM of adjacent segment in group A increased significantly at last follow-up ( t=2.318, P=0.026). There was significant difference in ROM of adjacent segment between the two groups ( P<0.05). Conclusion: The mid-term effectiveness of Coflex interspinous dynamic internal fixation combined with spinal fusion is similar to that of simple decompression fusion. For those patients whose adjacent segments of the responsible segments have degeneration but have no symptoms or mild symptoms, this treatment can slow down the adjacent segment degeneration.

Simultaneous quantitative detection of viable Escherichia coli O157:H7, Cronobacter spp., and Salmonella spp. using sodium deoxycholate-propidium monoazide with multiplex real-time PCR.

Escherichia coli O157:H7, Cronobacter spp., and Salmonella spp. are common food-borne pathogens in milk that may cause serious diseases. In the present study, we established a simple, rapid, and specific method to simultaneously detect viable E. coli O157:H7, Cronobacter spp., and Salmonella spp. in milk. Three specific genes, fliC from E. coli O157:H7, cgcA from Cronobacter spp., and invA from Salmonella spp., were selected and used to design primers and probes. False-positive results were eliminated with the use of a combined sodium deoxycholate (SD) and propidium monoazide (PMA) treatment. Using the optimized parameters, this SD-PMA treatment combined with multiplex real-time PCR (SD-PMA-mRT-PCR) detected E. coli O157:H7, Cronobacter spp. and Salmonella spp. respectively, at 102 cfu/mL in pure culture or artificially spiked skim milk samples. A reasonable recovery rate (from 100 to 107%) for detection of viable bacteria using the SD-PMA-mRT-PCR assay was obtained in the presence of dead bacteria at 107 cfu/mL. The SD-PMA-mRT-PCR method developed in this study can accurately detect and monitor combined contamination with E. coli O157:H7, Cronobacter spp., and Salmonella spp. in milk and milk products.