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Summary Rock pore structure and capillary number determine gas-water flow through limestone. Limestone is of key importance for natural gas production and gas storage; however, there is still limited direct evidence on the precise influence of the pore space morphology (vugs, fractures, and homogeneous pore matrix) and capillary number. Here, we thus studied gas-water flow patterns in various limestones via in-situ X-ray microtomography combined with numerical flow simulations. Pore structure heterogeneity significantly affected the fluid migration path. Gas flowed rapidly through large pores and vugs but flowed slowly through microfractures. In contrast, water flowed through microfractures and small pores but did not enter large pores at low capillary number conditions. Water flow simulations [performed for different capillary numbers directly on microcomputed tomography (μ-CT) images] demonstrated that snap-off and dead-end corners controlled the distribution of residual gas, consistent with the experiments. In addition, the simulations showed that less residual gas distributed around dead-end corners at a low capillary number, and a part of the residual gas can be displaced by increasing viscous forces. Moreover, a power-low relationship between gas cluster volume and surface area was observed, and the gas cluster size distribution could also be fitted with a power-law correlation. In all types of tested limestones, the power-law exponents (p ≈ 0.77, τ ≈ 0.86) were lower than that predicted by simple percolation theory (which predicts p ≈ 1, τ ≈ 2.189). There was evidence that a simple percolation model was unlikely to provide reliable predictions in homogeneous porous media, and we further extended the application scope of this conclusion to heterogeneous porous media. This work therefore provides fundamental data and improves fundamental understanding of gas-water flow through limestones and aids in the further advancements of improved hydrocarbon recovery and gas storage in limestone reservoirs.

1,6-Hexanediol (HDO) is an important precursor in the polymer industry. The current industrial route to produce HDO involves energy intensive and hazardous multistage (four-pot–four-step) chemical reactions using cyclohexane (CH) as the starting material, which leads to serious environmental problems. Here, we report the development of a biocatalytic cascade process for the biotransformation of CH to HDO under mild conditions in a one-pot–one-step manner. This cascade biocatalysis operates by using a microbial consortium composed of three E. coli cell modules, each containing the necessary enzymes. The cell modules with assigned functions were engineered in parallel, followed by combination to construct E. coli consortia for use in biotransformations. The engineered E. coli consortia, which contained the corresponding cell modules, efficiently converted not only CH or cyclohexanol to HDO, but also other cycloalkanes or cycloalkanols to related dihydric alcohols. In conclusion, the newly developed biocatalytic process provides a promising alternative to the current industrial process for manufacturing HDO and related dihydric alcohols.

A novel food packaging film was developed by incorporating a tea polyphenols-loaded pullulan/trehalose (TP@Pul/Tre) into a composite film with ultrasound-assisted treatment of dual-frequency (20/35 kHz, 40 W/L) for 15 min to assess the physicochemical and mechanical properties of a composite film. The optimized ultrasound-assisted significantly increases elongation at break, tensile strength, and improves the composite film's UV/water/oxygen barrier properties. Structure analysis using attenuated total reflectance-Fourier transform infrared, X-ray diffraction and thermal stability revealed that these improvements were achieved through ultrasound-enhanced H-bonds, more ordered molecular arrangements, and good intermolecular compatibility. Besides, the ultrasound-assisted TP@Pul/Tre film has proven to have good antibacterial performance against Escherichia coli and Staphylococcus aureus, with approximately 100 % lethality at 4 h and 8 h, respectively. Moreover, the ultrasound-assisted TP@Pul/Tre film effectively delayed moisture loss, oxidative browning, decay, and deterioration in fresh-cut apples and pears, thereby extending their shelf life. Thus, ultrasound has proved to be an effective tool for improving the quality of food packaging films, with a wide range of applications.

This study developed two novel food packaging films, oat protein/pullulan (Op/Pul) and Nisin-loaded oat protein/pullulan (Nis@Op/Pul) films. Ultrasound was introduced to improve its mechanical, structural and physicochemical properties. The Op/Pul film has lower light transmittance, water vapour and oxygen permeability (OP) and improved film uniformity than pure oat protein and pullulan film. The addition of Nisin led to a significant decrease in the composite films' transparency, moisture content, and total soluble matter (TSM). The ultrasound treatment significantly increased the elongation at break and transparency of Nis@Op/Pul film by 18.37% and 8.03% and decreased its TSM and OP by 8.33% and 2.78%, respectively, compared to the conventional method. The structure analysis shows ultrasound enhances intermolecular hydrogen bonding, reduces the crystallinity and formed a more regular, uniform surface. Moreover, the Nis@Op/Pul film prepared by ultrasound treatment could effectively delay the decay and deterioration of fresh strawberries and prolong their shelf life.

In this work, we derived a mathematical model for spontaneous imbibition in a Y-shaped branching network model. The classic Lucas–Washburn equation was used for modeling the imbibition process occurring in the Y-shape model. Then, a mathematical model for the Newtonian fluid’s imbibition was derived to reveal the relationship between dimensionless imbibition time and length ratio, radius ratio, and wetting strength. The dimensionless imbibition time in the model was adopted to compare with that of the capillary bundle model. Different length and radius ratios were considered in the adjacent two-stage channels, and different wettabilities were considered in the different branches. The optimal radius ratio, length ratio, and wetting strength were calculated under the condition of the shortest imbibition time. In addition, the shortest dimensionless imbibition time of the three-stage Y-shaped branching network model was calculated when the wettability changes randomly. The results indicate that the imbibition time changed mostly when the wettability of the second branch changed, and the second branch was the most sensitive to wettability in the model.

Increasing numbers of evidences have demonstrated that microRNAs (miRNAs) play an important role in osteosarcoma (OS) cell functions. MiR-501-3p has been reported to play an important role in several types of tumors, including prostate cancer and hepatocellular carcinoma. However, the biological function and potential mechanism of miR-501-3p in OS have not been well investigated until now. Here, we analyzed the expression of miR-501-3p in OS tissues and cell lines and its clinical significance in OS patients. Quantitative reverse transcription PCR showed miR-501-3p was significantly up-regulated in OS tissues and cell lines. Up-regulated miR-501-3p expression was associated with TNM stage, distal metastasis and worse prognosis in OS patients. MiR-501-3p knockdown and overexpression were achieved by miR-501-3p inhibitor and mimics transfection, respectively. CCK-8, colony formation and transwell assays showed that miR-501-3p knockdown in U2OS and Saos-2 cells suppressed, while miR-501-3p overexpression in Saos-2 cells promoted cell proliferation, migration and invasion. Moreover, luciferase reporter assay supporting BCL7A was a target of miR-501-3p and its expression was increased by miR-501-3p inhibitor, but inhibited by miR-501-3p mimics. By performing rescue experiments, we further demonstrated that BCL7A was a downstream functional regulator involved in miR-501-3p promoting OS cell functions. In summary, our findings suggest that miR-501-3p targets BCL7A may provide novel therapeutic targets for the treatment of OS.

The chitosanase gene (btbchito) of Bacillus thuringiensis BMB171 was cloned and heterologously expressed in the yeast Pichia pastoris. After purification, about 300 mg of recombinant chitosanase was obtained from the 1-1 culture medium with a specific activity of 240 units/mg. Results determined by the combined use of thin layer chromatography (TLC) and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) showed that the chitooligosaccharides (COSs) obtained by chitosan (N-deacetylated by 70%, 80%, and 90%) hydrolysis by rBTBCHITO were comprised of oligomers, with degrees of polymerization (DP) mainly ranging from trimers to heptamers; high molecular weight chitopentaose, chitohexaose, and chitoheptaose were also produced. Hydrolysis products was also deduced using MS since the COSs (n) are complex oligosaccharides with various acetyl groups from one to two, so the non-acetyl COSs (GlcN)n and COSs with more acetyls (> 2) were not detected. The employment of this method in the production of high molecular weight COSs may be useful for various industrial and biological applications, and the activity of chitosanase has great significance in research and other applications.

Purpose The effect of bisphosphonate on postoperative bone mineral density (BMD) is rarely reported. In the present study, the BMD change in ipsilateral hip with and without application of bisphosphonate after primary total knee arthroplasty (TKA) was evaluated. Methods and analysis Sixty-eight patients undergone TKA due to severe osteoarthritis were enrolled in this study. According to the application of bisphosphonate, the patients were divided into treatment group and control group. The BMD of ipsilateral hip was measured preoperatively and at the 3rd, 6th,12th and 24th month after surgery. Independent t -test was conducted to compare BMD difference between two groups preoperatively and paired t -test was conducted to compare BMD at each time point. The significant difference was considered at P Results No statistical difference of preoperative BMD in femoral neck, wards triangle, greater trochanter and proximal femur between two groups was detected. BMD in all ranges of interest (ROIs) in the control group reached the lowest level at the 12th month after surgery and increased slightly at the 24th month after surgery. The BMD in all ROIs in the treatment group revealed a slight fluctuation around the baseline level. Conclusion The application of bisphosphonate after TKA contributes to the BMD postoperatively. Bisphosphonate is helpful in reducing complications caused by osteoporosis after TKA as well as the recovery of patients. Therefore, patients can benefit from the application of bisphosphonate.

Toward an understanding of the protein interaction network of the human liver An extensive interaction network of human liver-expressed proteins is described, composed of 3484 interactions among 2582 proteins. Proteins associated with liver disease tend to be central and highly connected in the network., Proteome-scale protein interaction maps are available for many organisms, ranging from bacteria, yeast, worms and flies to humans. These maps provide substantial new insights into systems biology, disease research and drug discovery. However, only a small fraction of the total number of human protein–protein interactions has been identified. In this study, we map the interactions of an unbiased selection of 5026 human liver expression proteins by yeast two-hybrid technology and establish a human liver protein interaction network (HLPN) composed of 3484 interactions among 2582 proteins. The data set has a validation rate of over 72% as determined by three independent biochemical or cellular assays. The network includes metabolic enzymes and liver-specific, liver-phenotype and liver-disease proteins that are individually critical for the maintenance of liver functions. The liver enriched proteins had significantly different topological properties and increased our understanding of the functional relationships among proteins in a liver-specific manner. Our data represent the first comprehensive description of a HLPN, which could be a valuable tool for understanding the functioning of the protein interaction network of the human liver.

The clinical value of cardiac color ultrasound and cardiac troponin T (cTnT) combined with dynamic electrocardiogram in evaluating the treatment effect on acute pulmonary embolism (APE) was investigated. Ninety-eight patients with APE treated in Jinan Zhangqiu District Hospital of TCM from March 2016 to February 2017 were selected, and they were examined via the cardiac color ultrasound, cTnT and dynamic electrocardiogram to evaluate the clinical value in the treatment of pulmonary embolism. Electrocardiogram showed that there was no significant difference in the poor prognosis rate between right ventricular strain group and non-right ventricular strain group (P>0.05); the poor prognosis rate in cTnT positive group was significantly higher than that in cTnT negative group (P0.9, while that detected via dynamic electrocardiogram was

The sequence of an endo-chitosanase gene (CSN) from Aspergillus fumigatus was optimized based on the preferred codons of Pichia pastoris and synthesized in vitro through overlapping PCR (CSN-P). The gene was cloned into a yeast expression vector, pHBM905A, and secretorily expressed in Pichia pastoris GS115. The yield of CSN-P reached ~3 mg/ml with a high-density fermentation in a 14 l fermenter and the enzyme activity was ~25,000 U/ml. The enzyme had half-lives of 2.5 h at 80°C, 1 h at 90°C and 32 min at 100°C. It retained 70% activity after incubation with 10 M urea at room temperature for 30 min. This enzyme was used for a large-scale preparation of oligosaccharides: 3 g enzyme converted 200 kg chitosan into oligosaccharides in 24 h at 60°C.

Study design: Chronic strained lumbar disc herniation (LDH) cases were classified into bulging LDH, herniated LDH and prolapse LDH types according to imaging examination, and vertebrae disruptions were evaluated. Cytokines derived from the nucleus pulposus cells were detected, and their effects on osteoclastogenesis, as well as the mechanisms involved, were studied via an in vitro osteoclast differentiation system. Objective: To clarify the mechanisms of lumbar vertebrae resorption induced by lumbar herniation. Summary and background data: Chronic strained lumbar disc herniation induced vertebrae erosion exacerbates quality of patients’ life and clinical outcome. Although nucleus pulposus cells derived cytokines were reported to play an important role in this pathogenesis, the fundamental mechanisms underlying this process are still unclear. Methods: Chronic strained lumbar disc herniation patients were diagnosed with CT scan and T2-weighted magnetic resonance imaging. RNA was extracted from 192 surgical specimens of the herniated lumbar disc and 29 surgical excisions of the lumbar disc from spinal injury patients. The expressions of osteoclastogenesis related cytokines and chemokines were examined using real time PCR. Monocytes were induced into osteoclast with M-CSF and RANKL in vitro, while the IGF-1 and MCP-1 were added into the differentiation procedure in order to evaluate the effects and explore the molecular mechanisms. Results: Vertebrae erosion had a positive relationship with lumbar disc herniation severity types. In all of the osteoclastogenesis related cytokines, the IGF-1 and MCP-1 were the most highly expressed in the nucleus pulposus cells. IGF-1 enhances activation of NF-kB signaling directly, but MCP-1 upregulated the expression of RANK, so that enhanced cellular sensitivity to RANKL resulted in increasing osteoclastogenesis and activity. Conclusion: Lumbar herniation induced overexpression of IGF-1 and MCP-1 in nucleus pulposus cells aggravated vertebral erosions. Hence, this study suggests that targeting osteoclastogenesis related cytokines has potential clinical significance in the treatment of lumbar disc herniation patients.