[Construction of a novel tissue engineered meniscus scaffold based on low temperature deposition three-dimenisonal printing technology].

Objective: To investigate the construction of a novel tissue engineered meniscus scaffold based on low temperature deposition three-dimenisonal (3D) printing technology and evaluate its biocompatibility. Methods: The fresh pig meniscus was decellularized by improved physicochemical method to obtain decellularized meniscus matrix homogenate. Gross observation, HE staining, and DAPI staining were used to observe the decellularization effect. Toluidine blue staining, safranin O staining, and sirius red staining were used to evaluate the retention of mucopolysaccharide and collagen. Then, the decellularized meniscus matrix bioink was prepared, and the new tissue engineered meniscus scaffold was prepared by low temperature deposition 3D printing technology. Scanning electron microscopy was used to observe the microstructure. After co-culture with adipose-derived stem cells, the cell compatibility of the scaffolds was observed by cell counting kit 8 (CCK-8), and the cell activity and morphology were observed by dead/live cell staining and cytoskeleton staining. The inflammatory cell infiltration and degradation of the scaffolds were evaluated by subcutaneous experiment in rats. Results: The decellularized meniscus matrix homogenate appeared as a transparent gel. DAPI and histological staining showed that the immunogenic nucleic acids were effectively removed and the active components of mucopolysaccharide and collagen were remained. The new tissue engineered meniscus scaffolds was constructed by low temperature deposition 3D printing technology and it had macroporous-microporous microstructures under scanning electron microscopy. CCK-8 test showed that the scaffolds had good cell compatibility. Dead/live cell staining showed that the scaffold could effectively maintain cell viability (>90%). Cytoskeleton staining showed that the scaffolds were benefit for cell adhesion and spreading. After 1 week of subcutaneous implantation of the scaffolds in rats, there was a mild inflammatory response, but no significant inflammatory response was observed after 3 weeks, and the scaffolds gradually degraded. Conclusion: The novel tissue engineered meniscus scaffold constructed by low temperature deposition 3D printing technology has a graded macroporous-microporous microstructure and good cytocompatibility, which is conducive to cell adhesion and growth, laying the foundation for the in vivo research of tissue engineered meniscus scaffolds in the next step.

Robotic-assisted differential total knee arthroplasty with patient-specific implants: surgical techniques and preliminary results.

OBJECTIVE: In total knee arthroplasty (TKA), achieving soft-tissue balance while retaining acceptable lower limb alignment is sometimes difficult and may lead to patient dissatisfaction. Theoretically, patient-specific implants can bring great benefits, while the lack of precise surgical tools may hinder the improvement of outcomes. The objective of this study was to illustrate surgical techniques and evaluate kinematics and early clinical outcomes of robotic-assisted TKA using patient-specific implants. METHODS: Based on preoperative CT scan, femoral and tibial components were 3D printed. Medial and lateral tibial liners were separate with different thicknesses, posterior slopes and conformity. TiRobot Recon Robot was used for surgery, and was armed with smart tools that quantify gap, force and femoral-tibial track. We collected data on demographics, intraoperative gap balance and femoral-tibial motion. In the follow-up, we evaluated the range of motion, Visual Analogue Scale (VAS), forgotten joint score (FJS), Knee injury and Osteoarthritis Outcome Score, Joint Replacement (KOOS, JR) score. Radiological data were also harvested. RESULTS: Fifteen patients (17 knees) were enrolled with a mean age of 64.6 ± 6.4 (53-76) years. In 5 knees, we used symmetric tibial liners, the rest were asymmetric. After surgery, the average alignment was 1.6 ± 2.0 (-3-5) degrees varus. The average follow-up lasted 6.7 ± 4.2 (1-14) months. The mean visual analogue scale was 0.8 ± 0.7 (0-2), FJS was 62.4 ± 25.3 (0-87), KOOS was 86.5 ± 9.4 (57-97). 11 patients were "very satisfied", 3 were "satisfied" with the result, and one patient was neutral due to restricted extension and unsatisfactory rehabilitation at five months' follow-up. CONCLUSIONS: With patient-specific implants and robotics, TKA could be performed by a mathematical way, which was dubbed a "differential" TKA. Intraoperative kinematics was excellent in terms of gap-force balancing and femoral-tibial relative motion. Preliminary clinical outcomes were overall satisfactory.

SlPHL1 positively modulates acid phosphatase in response to phosphate starvation by directly activating the genes SlPAP10b and SlPAP15 in tomato.

Increased acid phosphatase (APase) activity is a prominent feature of tomato (Solanum lycopersicum) responses to inorganic phosphate (Pi) restriction. SlPHL1, a phosphate starvation response (PHR) transcription factor, has been identified as a positive regulator of low Pi (LP)-induced APase activity in tomato. However, the molecular mechanism underlying this regulation remains to be elucidated. Here, SlPHL1 was found to positively regulate the LP-induced expression of five potential purple acid phosphatase (PAP) genes, namely SlPAP7, SlPAP10b, SlPAP12, SlPAP15, and SlPAP17b. Furthermore, we provide evidence that SlPHL1 can stimulate transcription of these five genes by binding directly to the PHR1 binding sequence (P1BS) located on their promoters. The P1BS mutation notably weakened SlPHL1 binding to the promoters of SlPAP7, SlPAP12, and SlPAP17b but almost completely abolished SlPHL1 binding to the promoters of SlPAP10b and SlPAP15. As a result, the transcriptional activation of SlPHL1 on SlPAP10b and SlPAP15 was substantially diminished. In addition, not only did transient overexpression of either SlPAP10b or SlPAP15 in tobacco leaves increase APase activity, but overexpression of SlPAP15 in Arabidopsis and tomato also increased APase activity and promoted plant growth. Subsequently, two SPX proteins, SlSPX1 and SlSPX4, were shown to physically interact with SlPHL1. Moreover, SlSPX1 inhibited the transcriptional activation of SlPHL1 on SlPAP10b and SlPAP15 and negatively regulated the activity of APase. Taken together, these results demonstrate that SlPHL1-mediated LP signaling promotes APase activity by activating the transcription of SlPAP10b and SlPAP15, which may provide valuable insights into the mechanisms of tomato response to Pi-limited stress.

Mutation of OsNRAMP5 reduces cadmium xylem and phloem transport in rice plants and its physiological mechanism.

Natural resistance associated macrophage protein 5 (NRAMP5) is a key transporter for cadmium (Cd) uptake by rice roots; however, the effect of OsNRAMP5 on Cd translocation and redistribution in rice plants remains unknown. In this study, an extremely low Cd-accumulation mutant (lcd1) and wild type (WT) plants were utilized to investigate the effect of OsNRAMP5 mutation on Cd translocation and redistribution via the xylem and phloem and its possible physiological mechanism using field, hydroponic and isotope-labelling experiments. The results showed that OsNRAMP5 mutation reduced xylem and phloem transport of Cd, due to remarkably lower Cd translocation from roots to shoots and from the leaves Ⅰ-Ⅲ to their corresponding nodes, as well as lower Cd concentrations in xylem and phloem sap of lcd1 compared to WT plants. Mutation of OsNRAMP5 reduced Cd translocation from roots to shoots in lcd1 plants by increasing Cd deposition in cellulose of root cell walls and reducing OsHMA2-and OsCCX2-mediated xylem loading of Cd, and the citric acid- and tartaric acid-mediated long-distance xylem transport of Cd. Moreover, OsNRAMP5 mutation inhibited Cd redistribution from flag leaves to nodes and panicles in lcd1 plants by increasing Cd sequestration in cellulose and vacuoles, and decreasing OsLCT1-mediated Cd phloem transport in flag leaves.

Spatial and variety distributions, risk assessment, and prediction model for heavy metals in rice grains in China.

In this study, 6229 brown rice grains from three major rice-producing regions were collected to investigate the spatial and variety distributions of heavy metals in rice grains in China. The potential sources of heavy metals in rice grains were identified using the Pearson correlation matrix and principal component analysis, and the health risks of dietary exposure to heavy metals via rice consumption were assessed using the hazard index (HI) and total carcinogenic risk (TCR) method, respectively. Moreover, 48 paired soil and rice samples from 11 cities were collected to construct a predicting model for Cd accumulation in rice grains using the multiple linear stepwise regression analysis. The results indicated that Cd and Ni were the main heavy metal pollutants in rice grains in China, with approximately 10% of samples exceeding their corresponding maximum allowable limits. The Yangtze River basin had heavier pollution of heavy metals than the Southeast Coastal Region and Northeast Plain, and the indica rice varieties had higher heavy metal accumulation abilities compared with the japonica rice. The Cu, Pb, and Cd mainly originated from anthropogenic sources, while As, Hg, Cr, and Ni originated from both natural and anthropogenic sources. The mean HI and TCR values of dietary exposure to heavy metals via rice consumption ranged from 2.92 to 4.31 and 9.74 × 10-3 to 1.44 × 10-2, respectively, much higher than the acceptable range, and As and Ni were the main contributor to the HI and TCR for Chinese adults and children, respectively. The available Si (ASi), total Cd (TCd), available Mo (AMo), and available S (AS) were the main soil factors determining grain Cd accumulation. A multiple linear stepwise regression model was constructed based on ASi, TCd, AMo, and AS in soils with good accuracy and precision, which could be applied to predict Cd accumulation in rice grains and guide safe rice production in contaminated paddy fields.

Using sequential bone cutting to titrate soft tissue balance in total knee arthroplasty effectively minimizes soft tissue release.

BACKGROUND: Achieving soft tissue balance while maintaining limb alignment within acceptable boundaries is crucial for successful total knee arthroplasty (TKA). We proposed a sequential bone cutting (SBC) technique to titrate the soft tissue balance in robot-assisted TKA to achieve the desired balance with minimum soft tissue release. METHODS: In total, 106 robot-assisted TKAs using the SBC technique were included. The preoperative hip-knee-ankle angle (HKA) was < 10° in 76 and ≥ 10° in 30 knees. The gaps were initially balanced with the help of the pre-resection balancing provided by the robotic system. Soft tissue balance and alignment were quantitatively measured after the initial bone cutting and final bone cutting. Additional adjustments (bone recuts and soft tissue releases) required to address soft tissue imbalance after initial bone cutting were recorded. The frequencies of soft tissue releases, soft tissue balance, and resultant alignment ≤ 3° were compared between non-severe (HKA < 10°) and severe deformity (HKA ≥ 10°) groups. RESULTS: Soft tissue balance was achieved in 45 knees (42.5%) after initial bone cutting and in 93 knees (87.7%) after final balancing. The postoperative alignment was within 3° from neutral in 87 knees (82.1%) and 3-5° in 17 knees (16.0%). For unbalanced knees (n = 61) after initial bone cutting, soft tissue release was avoided by SBC in 37 knees (60.7%) and was deemed necessary in 24 knees (39.3%). Soft tissue release was more likely to be avoided in the non-severe deformity cohort (86.8% [33 of 38]) than in the severe deformity cohort (17.4% [4 of 23]; p < 0.001). The non-severe deformity cohort showed a significantly higher rate of resultant alignment ≤ 3° from neutral than the severe deformity cohort (90.8% vs. 60.0%; p < 0.001). CONCLUSION: Pre-resection balancing is inappropriate to ensure soft tissue balance. The SBC technique is effective in minimizing soft tissue release while maintaining overall alignment within acceptable boundaries.

Effects of Starch Molecular Structure and Physicochemical Properties on Eating Quality of Indica Rice with Similar Apparent Amylose and Protein Contents.

It is important to clarify the effects of starch fine structure and protein components on the eating quality of indica rice. In this study, seven indica rice varieties with similar apparent amylose content (AAC) and protein content (PC) but different sensory taste values were selected and compared systematically. It was found that except for AAC and PC, these varieties showed significant differences in starch molecular structure and protein components. Compared with rice varieties with a low sensory taste value, varieties with a higher sensory taste value showed significantly lower amylose and higher amylopectin short chains (degree of polymerization 6-12) content; the protein component showed that the varieties with good taste value had higher albumin and lower globulin and glutelin content (p < 0.05). Rice varieties with lower AC, globulin, and glutelin content, as well as a higher content of albumin and amylopectin short chains, resulted in a higher swelling factor, peak viscosity, breakdown value, and ratio of hardness to stickiness, in which condition cooked rice showed a higher sensory taste value. Moreover, this study indicated that rice varieties with a higher content of albumin and amylopectin short chains were conducive to the good appearance of cooked rice. This study lays the foundation for the taste evaluation of good-tasting indica rice.

Effects of different storage containers on the flavor characteristics of Jiangxiangxing baijiu.

Storage is a key factor controlling the quality of Jiangxiangxing baijiu, and storage time and the type of storage container play crucial roles in shaping the baijiu's distinct flavor. To investigate the influence of storage containers on the flavor characteristics of Jiangxiangxing baijiu, the sensory qualities, flavor components, and metal ions of Jiangxiangxing baijiu were measured during 24 months of storage in a pottery jar or a stainless steel tank. The results showed that Jiangxiangxing baijiu preserved in a pottery jar was superior to that stored in a stainless steel tank. A total of 96 flavor substances were detected, and 17 key flavor characteristic substances were screened by combining the results of odor activity values (OAV) and orthogonal partial least squares-discriminant analysis (OPLS-DA). A correlation heat map and redundancy analysis (RDA) showed that aluminum, cadmium, iron, cobalt, magnesium, potassium, and copper ions promoted the formation of key characteristic substances including diethoxymethane, lactic acid, 2,3-dimethyl-5-ethylpyrazine, 1-hexanol, and 2-methyl-1-propanol. Overall, the results show that 24-month pottery jar storage can promote the flavor quality of Jiangxiangxing baijiu. This study established a theoretical foundation to select the appropriate storage conditions and control the flavor quality of Jiangxiangxing baijiu.

Dynamic Changes in Volatiles, Soluble Sugars, and Fatty Acids in Glutinous Rice during Cooking.

Cooking is an important process before rice is consumed and constitutes the key process for rice flavor formation. In this paper, dynamic changes in aroma- and sweetness-related compounds were tracked during the entire cooking process (including washing with water, presoaking, and hydrothermal cooking). The volatiles, fatty acids, and soluble sugars in raw rice, washed rice, presoaked rice, and cooked rice were compared. After being washed with water, the total volatiles decreased while aldehydes and unsaturated fatty acids increased. Meanwhile, oligosaccharides decreased and monosaccharides increased. The changes in fatty acids and soluble sugars caused by the presoaking process were similar to those in the water-washing process. However, different changes were observed for volatiles, especially aldehydes and ketone. After hydrothermal cooking, furans, aldehydes, alcohols, and esters increased while hydrocarbons and aromatics decreased. Moreover, all fatty acids increased; among these, oleic acids and linoleic acid increased most. Unlike with washing and presoaking, all soluble sugars except fructose increased after hydrothermal cooking. Principal component analysis showed that cooked rice possessed a volatile profile that was quite different from that of uncooked rice, while washed rice and presoaked rice possessed similar volatile profiles. These results indicated that hydrothermal cooking is the pivotal process for rice flavor formation.

Hierarchical porous ECM scaffolds incorporating GDF-5 fabricated by cryogenic 3D printing to promote articular cartilage regeneration.

BACKGROUND: In recent years, there has been significant research progress on in situ articular cartilage (AC) tissue engineering with endogenous stem cells, which uses biological materials or bioactive factors to improve the regeneration microenvironment and recruit more endogenous stem cells from the joint cavity to the defect area to promote cartilage regeneration. METHOD: In this study, we used ECM alone as a bioink in low-temperature deposition manufacturing (LDM) 3D printing and then successfully fabricated a hierarchical porous ECM scaffold incorporating GDF-5. RESULTS: Comparative in vitro experiments showed that the 7% ECM scaffolds had the best biocompatibility. After the addition of GDF-5 protein, the ECM scaffolds significantly improved bone marrow mesenchymal stem cell (BMSC) migration and chondrogenic differentiation. Most importantly, the in vivo results showed that the ECM/GDF-5 scaffold significantly enhanced in situ cartilage repair. CONCLUSION: In conclusion, this study reports the construction of a new scaffold based on the concept of in situ regeneration, and we believe that our findings will provide a new treatment strategy for AC defect repair.

STS loaded PCL-MECM based hydrogel hybrid scaffolds promote meniscal regeneration via modulating macrophage phenotype polarization.

Meniscus injury has a limited ability to heal itself and often results in the progression to osteoarthritis. After a meniscus injury, there is an obvious acute or chronic inflammatory response in the articular cavity, which is not conducive to tissue regeneration. M2 macrophages are involved in tissue repair and remodeling. Regenerative medicine strategies for tissue regeneration by enhancing the phenotypic ratio of M2 : M1 macrophages have been demonstrated in a variety of tissues. However, there are no relevant reports in the field of meniscus tissue regeneration. In this study, we confirmed that sodium tanshinone IIA sulfonate (STS) could transform macrophages from M1 to M2 polarization. STS protects meniscal fibrochondrocytes (MFCs) against the effects of macrophage conditioned medium (CM). Moreover, STS attenuates interleukin (IL)-1ß-induced inflammation, oxidative stress, apoptosis, and extracellular matrix (ECM) degradation in MFCs, possibly by inhibiting the interleukin-1 receptor-associated kinase 4 (IRAK4)/TNFR-associated factor 6 (TRAF6)/nuclear factor-kappaB (NF-κB) signaling pathway. An STS loaded polycaprolactone (PCL)-meniscus extracellular matrix (MECM) based hydrogel hybrid scaffold was fabricated. PCL provides mechanical support, the MECM based hydrogel provides a microenvironment conducive to cell proliferation and differentiation, and STS is used to drive M2 polarization and protect MFCs against the effects of inflammatory stimuli, thus providing an immune microenvironment conducive to regeneration. The results of subcutaneous implantation in vivo showed that hybrid scaffolds could induce M2 polarization in the early stage. In addition, the hybrid scaffolds seeded with MFCs could achieve good meniscus regeneration and chondroprotective effects in rabbits.

Engineering exosomes by three-dimensional porous scaffold culture of human umbilical cord mesenchymal stem cells promote osteochondral repair.

Improving the poor microenvironment in the joint cavity has potential for treating cartilage injury, and mesenchymal stem cell (MSC)-derived exosomes (MSC-Exos), which can modulate cellular behavior, are becoming a new cell-free therapy for cartilage repair. Here, we used acellular cartilage extracellular matrix (ACECM) to prepare 3D scaffolds and 2D substrates by low-temperature deposition modeling (LDM) and tape casting. We aimed to investigate whether MSC-Exos cultured on scaffolds of different dimensions could improve the poor joint cavity microenvironment caused by cartilage injury and to explore the related mechanisms. In vitro experiments showed that exosomes derived from MSCs cultured on three-dimensional (3D) scaffolds (3D-Exos) had increased efficiency. In short-term animal experiments, compared with exosomes derived from MSCs cultured in a two-dimensional (2D) environment (2D-Exos), 3D-Exos had a stronger ability to regulate the joint cavity microenvironment. Long-term animal studies confirmed the therapeutic efficacy of 3D-Exos over 2D-Exos. Thus, 3D-Exos were applied in the rat knee osteochondral defect model after adsorption in the micropores of the scaffold and combined with subsequent articular cavity injections, and they showed a stronger cartilage repair ability. These findings provide a new strategy for repairing articular cartilage damage. Furthermore, miRNA sequencing indicated that the function of 3D-Exos may be associated with high expression of miRNAs. Thus, our study provides valuable insights for the design of 3D-Exos to promote cartilage regeneration.

Multifaceted Catalytic ROS-Scavenging via Electronic Modulated Metal Oxides for Regulating Stem Cell Fate.

Developing reactive oxygen species (ROS)-scavenging nanostructures to protect and regulate stem cells has emerged as an intriguing strategy for promoting tissue regeneration, especially in trauma microenvironments or refractory wounds. Here, an electronic modulated metal oxide is developed via Mn atom substitutions in Co3 O4 nanocrystalline (Mn-Co3 O4 ) for highly efficient and multifaceted catalytic ROS-scavenging to reverse the fates of mesenchymal stem cells (MSCs) in oxidative-stress microenvironments. Benefiting from the atomic Mn-substitution and charge transfer from Mn to Co, the Co site in Mn-Co3 O4 displays an increased ratio of Co2+ /Co3+ and improved redox properties, thus enhancing its intrinsic and broad-spectrum catalytic ROS-scavenging activities, which surpasses most of the currently reported metal oxides. Consequently, the Mn-Co3 O4 can efficiently protect the MSCs from ROS attack and rescue their functions, including adhesion, spreading, proliferation, and osteogenic differentiation. This work not only establishes an efficient material for catalytic ROS-scavenging in stem-cell-based therapeutics but also provides a new avenue to design biocatalytic metal oxides via modulation of electronic structure.

Development of Certified Reference Materials for the Determination of Apparent Amylose Content in Rice.

Apparent amylose content (AAC) is one of the most important parameters in rice quality evaluation. In this study, four rice reference materials used to test rice AAC were developed. The AAC of rice reference materials were measured by a spectrophotometric method with a defatting procedure, calibrated from potato amylose and waxy rice amylopectin at the absorption wavelengths of 620 and 720 nm. Homogeneity test (n = 20) was judged by F-test based on the mean squares of among and within bottles, and short- and long-term stability monitoring was performed by T-test to check if there was significant degradation at the delivery temperature of under 40 °C (14 days) and at 0-4 °C storage condition (18 months), respectively. After joint evaluation by ten laboratories, Dixion and Cochran statistical analyses were presented. The expanded uncertainties were calculated based on the uncertainty of homogeneity, short- and long-term stability, and inter-laboratory validation containing factor k = 2. It found that the four reference materials were homogenous and stable, and had the AAC (g/100 g, k = 2) of 2.96 ± 1.01, 10.68 ± 0.66, 17.18 ± 1.04, and 16.09 ± 1.29, respectively, at 620 nm, and 1.46 ± 0.49, 10.44 ± 0.56, 16.82 ± 0.75, and 24.33 ± 0.52, respectively, at 720 nm. It was indicated that 720 nm was more suitable for the determination of rice AAC with lower uncertainties. The determinations of the AAC of 11 rice varieties were carried out by two methods, the method without defatting and with calibration from the four rice reference materials and the method with a defatting procedure and calibrating from potato amylose and waxy rice amylopectin. It confirmed that the undefatted rice reference materials could achieve satisfactory results to test the rice samples with the AAC ranging from 1 to 25 g/100 g. It would greatly reduce the time cost and improve testing efficiency and applicability, and provide technical support for the high-quality development of the rice industry.

[Determination of three oryzanols in rice by mixed-mode solid-phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry].

Rice is a major dietary staple in many communities owing to its high nutritional value and characteristic aroma. Oryzanol, a mixture of ferulic acid esters of triterpene alcohols and phytosterols, is a major group of phytochemicals found in rice. 24-Methylenecycloartanyl ferulate (24MCA-FA), cycloartenyl ferulate (CA-FA), and campestanyl ferulate (Camp-FA) have been identified as the primary components of oryzanol. At present, for the quantification of oryzanol in rice and rice products, UV spectroscopy or high performance liquid chromatography (HPLC) is widely employed. However, these methods cannot differentiate individual oryzanols, resulting in higher measured values. To extract oryzanol, methods including liquid-liquid extraction, acidulation extraction, and direct solvent extraction have been typically employed, as they do not require specific extraction instrumentation. However, there has been no systematic study on the direct solvent extraction and purification conditions of oryzanol in rice. In this study, a rapid and accurate analytical method based on HPLC-MS/MS and mixed-mode anion exchange (MAX) solid-phase extraction was established to determine the content of three oryzanols (24MCA-FA, CA-FA, and Camp-FA) in rice. The MS parameters, such as the collision energy of three ion pairs of each oryzanol, were optimized. Further, the chromatographic separation conditions and response intensities of the oryzanols in different mobile phases were compared. The effects of different pretreatment conditions on the extraction efficiency of the three oryzanols in rice samples and different purification conditions on their recovery were investigated. Combined with the external standard method, the three oryzanols in rice were successfully quantified. The results showed that the baseline separation and highest response for the three oryzanols were achieved using the Agilent Eclipse XDB-C8 chromatographic column (150 mm×2.1 mm, 3.5 µm) when methanol∶ acetonitrile in a 1∶1 ratio (v/v) and an aqueous solution of 5 mmol/L ammonium acetate were used as the mobile phases for gradient elution. The extraction rate of the three oryzanols was highest when using 2.5 g of the sample, adding 20 mL of methanol, soaking for 12 h, ultrasonicating at a temperature of 40 ℃ for 20 min, and centrifuging the extracted solutions at 4500 r/min for 10 min. The samples were purified by MAX, and the sample matrix effect was found to be lesser than 1.6%-10.8%. Under the optimum conditions, the calibration curves of the three oryzanols showed good linearity (correlation coefficients r2≥0.9983) within their respective linear ranges. The limits of detection were in the range of 0.5-1.0 µg/L, and limits of quantification were in the range of 2.0-3.5 µg/L. Accuracy and precision experiments were performed on rice samples spiked at three levels (2, 5, and 10 times the background concentration), with three replicates. The average recoveries of the three oryzanols ranged from 86.1% to 110.6%, and the relative standard deviations (RSDs) were between 0.9% and 3.2%. The method showed good performance when applied to the analysis of real samples. In conclusion, the developed method can determine the content of the three oryzanols in rice quickly and accurately, and can be used for the subsequent measurement of oryzanol compounds in rice.

Hydrogel composite scaffolds achieve recruitment and chondrogenesis in cartilage tissue engineering applications.

BACKGROUND: The regeneration and repair of articular cartilage remains a major challenge for clinicians and scientists due to the poor intrinsic healing of this tissue. Since cartilage injuries are often clinically irregular, tissue-engineered scaffolds that can be easily molded to fill cartilage defects of any shape that fit tightly into the host cartilage are needed. METHOD: In this study, bone marrow mesenchymal stem cell (BMSC) affinity peptide sequence PFSSTKT (PFS)-modified chondrocyte extracellular matrix (ECM) particles combined with GelMA hydrogel were constructed. RESULTS: In vitro experiments showed that the pore size and porosity of the solid-supported composite scaffolds were appropriate and that the scaffolds provided a three-dimensional microenvironment supporting cell adhesion, proliferation and chondrogenic differentiation. In vitro experiments also showed that GelMA/ECM-PFS could regulate the migration of rabbit BMSCs. Two weeks after implantation in vivo, the GelMA/ECM-PFS functional scaffold system promoted the recruitment of endogenous mesenchymal stem cells from the defect site. GelMA/ECM-PFS achieved successful hyaline cartilage repair in rabbits in vivo, while the control treatment mostly resulted in fibrous tissue repair. CONCLUSION: This combination of endogenous cell recruitment and chondrogenesis is an ideal strategy for repairing irregular cartilage defects.

The potential role of plasma membrane proteins in response to Zn stress in rice roots based on iTRAQ and PRM under low Cd condition.

Cd pollution had already caused serious threats to crop growth and development, food safety and human health, and become a potential agricultural and global environmental problem. Zn had been used to reduce Cd accumulation in soil and plants. Proteins located in plasma membrane (PM) played important roles in transferring stress signals in plants. To further elucidate how PM proteins modulated Zn/Cd transport under low-Cd condition, quantitative proteomics was employed to identify and verify the differentially expressed proteins (DEPs) and their biological functions at proteome level. A total of 4008 proteins were identified, and 332 DEPs (192 up and 140 down, fold >1.50 or <0.66, p < 0.01) were screened. Functional analysis showed that DEPs were mainly catalytic active and binding proteins, involved in glutathione metabolism, phenylpropanoid biosynthesis, etc. DEPs involved in ion transport played key roles in regulating transmembrane transport, resisting stress and alleviating toxicity of heavy metals to rice roots. DEPs were as the marker proteins in rice root responding to heavy metal stress. This study had important guiding significances for metal ions transport mechanism and screening of biomarkers responding to abiotic stress, and provided references for further researches underlying abiotic stress and detoxication in rice and other plants.

The immune microenvironment in cartilage injury and repair.

The ability of articular cartilage to repair itself is limited because it lacks blood vessels, nerves, and lymph tissue. Once damaged, it can lead to joint swelling and pain, accelerating the progression of osteoarthritis. To date, complete regeneration of hyaline cartilage exhibiting mechanical properties remains an elusive goal, despite the many available technologies. The inflammatory milieu created by cartilage damage is critical for chondrocyte death and hypertrophy, extracellular matrix breakdown, ectopic bone formation, and progression of cartilage injury to osteoarthritis. In the inflammatory microenvironment, mesenchymal stem cells (MSCs) undergo aberrant differentiation, and chondrocytes begin to convert or dedifferentiate into cells with a fibroblast phenotype, thereby resulting in fibrocartilage with poor mechanical qualities. All these factors suggest that inflammatory problems may be a major stumbling block to cartilage repair. To produce a milieu conducive to cartilage repair, multi-dimensional management of the joint inflammatory microenvironment in place and time is required. Therefore, this calls for elucidation of the immune microenvironment of cartilage repair after injury. This review provides a brief overview of: (1) the pathogenesis of cartilage injury; (2) immune cells in cartilage injury and repair; (3) effects of inflammatory cytokines on cartilage repair; (4) clinical strategies for treating cartilage defects; and (5) strategies for targeted immunoregulation in cartilage repair. STATEMENT OF SIGNIFICANCE: Immune response is increasingly considered the key factor affecting cartilage repair. It has both negative and positive regulatory effects on the process of regeneration and repair. Proinflammatory factors are secreted in large numbers, and necrotic cartilage is removed. During the repair period, immune cells can secrete anti-inflammatory factors and chondrogenic cytokines, which can inhibit inflammation and promote cartilage repair. However, inflammatory factors persist, which accelerate the degradation of the cartilage matrix. Furthermore, in an inflammatory microenvironment, MSCs undergo abnormal differentiation, and chondrocytes begin to transform or dedifferentiate into fibroblast-like cells, forming fibrocartilage with poor mechanical properties. Consequently, cartilage regeneration requires multi-dimensional regulation of the joint inflammatory microenvironment in space and time to make it conducive to cartilage regeneration.

Mapping and functional analysis of high-copper accumulation mutant oshc1 in rice.

Copper (Cu) is an essential but potentially toxic element in rice. Little is known about the mechanism of rice grain Cu accumulation. In this study, we identified a high copper accumulation in grain 1 (oshc1) mutant from the wild type indica rice cultivar 9311 (WT) mutant bank. Compared with those in WT, more Cu was shown to accumulate in the shoots of seedlings and the above-ground tissues except nodes although less total Cu content in oshc1. Further analysis showed that the mutant had an accelerated Cu transport ratio from roots to shoots and higher Cu concentration in xylem sap than WT. This phenomenon in oshc1 was controlled by a single recessive gene, which was identified as BGIOSGA007732, and named OsHMA4. The eight base frame-shift from 1021 to 1028 bp in the coding sequence of OsHMA4 led to a modification after the 341st amino acid and resulted in premature translation termination of OsHMA4 at the 377th amino acid. This may change the function of OsHMA4. Furthermore, the up-regulated OsCOPT7 and OsATX1 and down-regulated OsHMA4 probably decrease Cu compartmentalization in roots of oshc1. In summary, the frame-shift in OsHMA4 changes the function of OsHMA4 and the expression of genes relative to Cu transport in the mutant, which leads to more Cu transport upward and higher Cu accumulation in the rice grains. Moreover, oshc1 was more tolerance to Cu-shortage than WT, while more sensitive to Cu excess exposure than WT. However, RNA-Seq analysis shown that changes in transcription levels of genes in oshc1 involving in molecular function of ions binding and biological processes of cell wall organization and defense response to bio-stress. Which indicates that oshc1 is advantage to Cu limited condition than WT. This work reveals the mechanism of high Cu accumulation in the grains of oshc1 and provides a material to breed new cultivars with optimum levels of Cu in brown rice by crossing with other dominant varieties, which can be planted in different soils to ensure the yield and quality of rice.