3D printing technology and its combination with nanotechnology in bone tissue engineering.

With the graying of the world's population, the morbidity of age-related chronic degenerative bone diseases, such as osteoporosis and osteoarthritis, is increasing yearly, leading to an increased risk of bone defects, while current treatment methods face many problems, such as shortage of grafts and an incomplete repair. Therefore, bone tissue engineering offers an alternative solution for regenerating and repairing bone tissues by constructing bioactive scaffolds with porous structures that provide mechanical support to damaged bone tissue while promoting angiogenesis and cell adhesion, proliferation, and activity. 3D printing technology has become the primary scaffold manufacturing method due to its ability to precisely control the internal pore structure and complex spatial shape of bone scaffolds. In contrast, the fast development of nanotechnology has provided more possibilities for the internal structure and biological function of scaffolds. This review focuses on the application of 3D printing technology in bone tissue engineering and nanotechnology in the field of bone tissue regeneration and repair, and explores the prospects for the integration of the two technologies.

Accurate Spatio-Temporal Delivery of Nitric Oxide Facilitates the Programmable Repair of Avascular Dense Connective Tissues Injury.

Avascular dense connective tissues (e.g., the annulus fibrosus (AF) rupture, the meniscus tear, and tendons and ligaments injury) repair remains a challenge due to the "biological barrier" that hinders traditional drug permeation and limits self-healing of the injured tissue. Here, accurate delivery of nitric oxide (NO) to penetrate the "AF biological barrier" is achieved thereby enabling programmable AF repair. NO-loaded BioMOFs are synthesized and mixed in a modified polyvinyl alcohol and PCL-composited electrospun fiber membrane with excellent reactive oxygen species-responsive capability (LN@PM). The results show that LN@PM could respond to the high oxidative stress environment at the injured tissue and realize continuous and substantial NO release. Based on low molecular weight and lipophilicity, NO could penetrate through the "biological barrier" for accurate AF drug delivery. Moreover, the dynamic characteristics of the LN@PM reaction can be matched with the pathological microenvironment to initiate programmable tissue repair including sequential remodeling microenvironment, reprogramming the immune environment, and finally promoting tissue regeneration. This tailored programmable treatment strategy that matches the pathological repair process significantly repairs AF, ultimately alleviating intervertebral disc degeneration. This study highlights a promising approach for avascular dense connective tissue treatment through intelligent NO release, effectively overcoming "AF biological barriers" and programmable treatment.

Ubiquitous conservative interaction patterns between post-spliced introns and their mRNAs revealed by genome-wide interspecies comparison.

Introns, as important vectors of biological functions, can influence many stages of mRNA metabolism. However, in recent research, post-spliced introns are rarely considered. In this study, the optimal matched regions between introns and their mRNAs in nine model organism genomes were investigated with improved Smith-Waterman local alignment software. Our results showed that the distributions of mRNA optimal matched frequencies were highly consistent or universal. There are optimal matched frequency peaks in the UTR regions, which are obvious, especially in the 3'-UTR. The matched frequencies are relatively low in the CDS regions of the mRNA. The distributions of the optimal matched frequencies around the functional sites are also remarkably changed. The centers of the GC content distributions for different sequences are different. The matched rate distributions are highly consistent and are located mainly between 60% and 80%. The most probable value of the optimal matched segments is about 20 bp for lower eukaryotes and 30 bp for higher eukaryotes. These results show that there are abundant functional units in the introns, and these functional units are correlated structurally with all kinds of sequences of mRNA. The interaction between the post-spliced introns and their corresponding mRNAs may play a key role in gene expression.

A Composite-Type MEMS Pirani Gauge for Wide Range and High Accuracy.

To achieve a wide range and high accuracy detection of the vacuum level, for example, in an encapsulated vacuum microcavity, a composite-type MEMS Pirani gauge has been designed and fabricated. The Pirani gauge consists of two gauges of different sizes connected in series, with one gauge having a larger heat-sensitive area and a larger air gap for extending the lower measurable limit of pressure (i.e., the high vacuum end) and the other gauge having a smaller heat-sensitive area and a smaller air gap for extending the upper measurable limit. The high-resistivity titanium metal was chosen as the thermistor; SiNx was chosen as the dielectric layer, considering the factors relevant to simulation and manufacturing. By simulation using COMSOL Multiphysics and NI Multisim, a range of measurement of 2 × 10-2 to 2 × 105 Pa and a sensitivity of 52.4 mV/lgPa were obtained in an N2 environment. The performance of the fabricated Pirani gauge was evaluated by using an in-house made vacuum test system. In the test, the actual points of measurement range from 6.6 × 10-2 to 1.12 × 105 Pa, and the highest sensitivity is up to 457.6 mV/lgPa. The experimental results are better in the range of measurement, sensitivity, and accuracy than the simulation results. The Pirani gauge proposed in this study is simple in structure, easy to manufacture, and suitable for integration with other MEMS devices in a microcavity to monitor the vacuum level therein.

Examination of the microstructures of the lower cervical facet based on micro-computed tomography: A cadaver study.

The cervical facet has complicated 3D microstructures and inhomogeneities. The cervical facet joint, which also participates in the formation, plays a certain role in regulating and limiting the movement of the spine. Correct identification and evaluation of its microstructure can help in the diagnosis of orthopedic disease and predict early phases of fracture risk. To evaluate the safety of the cervical spine by measuring and analyzing the microstructures and morphometric parameters of bone trabeculae in the normal cervical facet with high-resolution 3D micro-computed tomography. Thirty-one sets of C3 to C7 lower cervical vertebrae (155 vertebrae) were scanned using micro-computed tomography. The morphological characteristics and direction of trabecular bone in the facet of the lower cervical vertebrae were observed by selecting and rebuilding the areas of interest, and the changes in the microstructure of the areas of interest were calculated to reveal the structural characteristics and weak areas. Images indicated an ossified center between the superior and inferior articular processes of the lower cervical spine. The cellular bone trabeculae of the articular process had complex reticular microstructures. The trabecular bone plate near the cortical bone was lamellar and relatively dense, and it extended around and transformed into a network structure, and then into the rod-shaped trabecular bone. The rod-shaped trabeculae converged with the plate-shaped trabeculae with only 1 to 2 layers surrounding the trabeculae cavity. Statistical results of the morphological parameters of the trabecular bone showed that trabecular bone volume fraction values were significantly higher for C7 than for C3 to C6 (P < .05). There were significant differences between C7 and C3 to C5 and between C6 and C4 in bone surface area/bone volume (P < .05). There was a significant difference between C7 and C3 to C6 in trabecular bone thickness values (P < .05). The degree of anisotropy value was significantly smaller for C3 than for C6 and C7 (P < .05). The changes in the C3 to C7 microstructure were summarized in this study. The loading capacity and stress of the C7 articular process tended to be limited, and the risk of injury tended to be higher for the C7 articular process.

Design of a High Sensitivity Pirani Gauge Based on Vanadium Oxide Film for High Vacuum Measurement.

We have designed a hot-plate-type micro-Pirani vacuum gauge with a simple structure and compatibility with conventional semiconductor fabrication processes. In the Pirani gauge, we used a vanadium oxide (VOx) membrane as the thermosensitive component, taking advantage of the high temperature coefficient of resistance (TCR) of VOx. The TCR value of VOx is -2%K-1∼-3%K-1, an order of magnitude higher than those of other thermal-sensitive materials, such as platinum and titanium (0.3%K-1∼0.4%K-1). On one hand, we used the high TCR of VOx to increase the Pirani sensitivity. On the other hand, we optimized the floating structure to decrease the thermal conductivity so that the detecting range of the Pirani gauge was extended on the low-pressure end. We carried out simulation experiments on the thermal zone of the Pirani gauge, the width of the cantilever beam, the material and thickness of the supporting layer, the thickness of the thermal layer (VOx), the depth of the cavity, and the shape and size. Finally, we decided on the basic size of the Pirani gauge. The prepared Pirani gauge has a thermal sensitive area of 130 × 130 µm2, with a cantilever width of 13 µm, cavity depth of 5 µm, supporting layer thickness of 300 nm, and VOx layer thickness of 110 nm. It has a dynamic range of 10-1~104 Pa and a sensitivity of 1.23 V/lgPa. The VOx Pirani was designed using a structure and fabrication process compatible with a VOx-based uncooled infrared microbolometer so that it can be integrated by wafer level. This work contains only our MEMS Pirani gauge device design, preparation process design, and readout circuit design, while the characterization and relevant experimental results will be reported in the future.

Structural Optimization and Mechanical Simulation of MEMS Thin-Film Getter-Heater Unit.

A MEMS thin-film getter-heater unit has been previously proposed for the vacuum packaging of a Micro-Electro-Mechanical System (MEMS) device, where the floating structure (FS) design is found to be obviously more power-efficient than the solid structure (SS) one by heat transfer capacity simulation. However, the mechanical strength of the FS is weaker than the SS by nature. For high temperature usage, the unit structure must be optimized in order to avoid fracture of the cantilever beam or film delamination due to strong excessive stress caused by heating. In this paper, COMSOL is used to simulate the stress and deformation of the MEMS thin-film getter-heater unit with the cantilever structure. By comparing various cantilever structures, it is found that a model with a symmetrically-shaped heater and edge-center-located cantilever model (II-ECLC model) is the most suitable. In this model, even when the structure is heated to about 600 °C, the maximum stress of the cantilever beam is only 455 MPa, much lower than the tensile strength of silicon nitride (Si3N4, 12 GPa), and the maximum deformation displacement is about 200 µm. Meanwhile, the interfacial stress between the getter and the insulating layer is 44 MPa, sufficiently lower than the adhesion strength between silicon nitride film and titanium film (400-1850 MPa). It is further found that both the stress of the cantilever structure and the interfacial stress between the getter and the insulating layer beneath increase linearly with temperature; and the deformation of the cantilever structure is proportional to its stress. This work gives guidance on the design of MEMS devices with cantilever structures and works in high temperature situations.

Water-fueled autocatalytic bactericidal pathway based on e-Fenton-like reactions triggered by galvanic corrosion and extracellular electron transfer.

Water is generally considered to be an undesirable substance in fuel system, which may lead to microbial contamination. The antibacterial strategies that can turn water into things of value with high disinfection efficacy have been urgently needed for fuel system. Here, we reveal a water-fueled autocatalytic bactericidal pathway comprised by bi-metal micro-electrode system, which can spontaneously produce reactive oxygen species (mainly H2O2 and O2•-) by the electron Fenton-like reaction in water medium without external energy., The respiratory chain component of bacteria and the galvanic corrosion on the coated metals were two electron sources in the system. The specific model of Ag-Ru water-fueled autocatalytic (WFA) microelectrode particles presents extremely high disinfection efficiency (>99.9999%) in less than one hour for three aerobic bacteria (Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis) in LB media and high disinfection efficiency for the anaerobic bacteria (Desulfovibrio alaskensis) in Postgate E media without natural light irradiation. Overall, the novel WFA Ag-Ru antibacterial material explored in this study has a high potential for sterilizing applications in fuel system and this work provides the potential for the development of non-chemical and water-based antibacterial materials, such as WFA Ag-Ru antibacterial coating on stainless steel.

C-type lectin domain-containing protein CLEC3A regulates proliferation, regeneration and maintenance of nucleus pulposus cells.

It is widely assumed that as connective tissue, the intervertebral disc (IVD) plays a crucial role in providing flexibility for the spinal column. The disc is comprised of three distinct tissues: the nucleus pulposus (NP), ligamentous annulus fibrous (AF) that surrounds the NP, and the hyaline cartilaginous endplates (CEP). Nucleus pulposus, composed of chondrocyte-like NP cells and its secreted gelatinous matrix, is critical for disc health and function. The NP matrix underwent dehydration accompanied by increasing fibrosis with age. The degeneration of matrix is almost impossible to repair, with the consequence of matrix stiffness and senescence of NP cells and intervertebral disc, suggesting the value of glycoproteins in extracellular matrix (ECM). Here, via database excavation and biological function screening, we investigated a C-type lectin protein, CLEC3A, which could support differentiation of chondrocytes as well as maintenance of NP cells and was essential to intervertebral disc homeostasis. Furthermore, mechanistic analysis revealed that CLEC3A could stimulate PI3K-AKT pathway to accelerate cell proliferation to further play part in NP cell regeneration.

Social exclusion, thwarted belongingness, and perceived burdensomeness: construct validity and psychometric properties of the Interpersonal Needs Questionnaire among patients with sexually transmitted infections in Shanghai, China.

BACKGROUND: Sexually transmitted infections (STIs) are a serious public health problem worldwide. Patients with STIs have a high rate of psychosocial problems and may perceive unmet interpersonal needs, which is considered a proximal and sufficient cause of suicidal thoughts and behaviors. The present study examined the construct validity and psychometric properties of the 15-item Interpersonal Needs Questionnaire among patients with STIs in Shanghai, China. METHODS: We recruited 910 patients with STIs (438 males and 472 females; mean age = 38.72, standard deviation [SD] = 13.034) from the Shanghai Skin Disease Hospital using accidental sampling. Baseline descriptive statistics were calculated using R 4.0.0, and a latent variable model was developed using Mplus 7.4. RESULTS: The construct validity results supported a latent variable measurement model with three distinct but related constructs (thwarted belongingness, perceived burdensomeness, and social exclusion) (p < 0.001, χ2/df = 2.475, root mean square error of approximation = 0.057, comparative fit index = 0.931, Tucker-Lewis index = 0.916, standardized root mean residual = 0.044). The Cronbach's α and McDonald's ω values were 0.849 and 0.767 for the total scale, 0.888 and 0.889 for perceived burdensomeness, 0.764 and 0.777 for social exclusion, and 0.892 and 0.893 for thwarted belongingness. Interpersonal needs were significantly associated with low self-esteem (r = 0.539), loneliness (r = 0.573), depression (r = 0.338), entrapment (r = 0.420), defeat (r = 0.579), and low perceived social support (r = 0.424). CONCLUSIONS: This was the first study to highlight social exclusion as a distinct but related dimension of interpersonal needs. This finding indicates that patients with STIs perceive high social exclusion. Therefore, health providers should consider the psychological status of these patients and implement strategies to support their integration into society.

Process Optimization and Performance Evaluation of TSV Arrays for High Voltage Application.

In order to obtain high-quality through-silicon via (TSV) arrays for high voltage applications, we optimized the fabrication processes of the Si holes, evaluated the dielectric layers, carried out hole filling by Cu plating, and detected the final structure and electric properties of the TSVs. The Si through-hole array was fabricated in an 8-inch Si substrate as follows: First, a blind Si hole array was formed by the Si deep reactive etching (DRIE) technique using the Bosch process, but with the largest width of the top scallops reduced to 540 nm and the largest notch elimidiameternated by backside grinding, which also opens the bottom ends of the Si blind holes and forms 500-µm-deep Si through holes. Then, the sidewalls of the Si holes were further smoothed by a combination of thermal oxidation and wet etching of the thermal oxide. The insulating capability of the dielectric layers was evaluated prior to metal filling by using a test kit. The metal filling of the through holes was carried out by bottom-up Cu electroplating and followed by annealing at 300 °C for 1 h to release the electroplating stress and to prevent possible large metal thermal expansion in subsequent high-temperature processes. The TSV arrays with different hole diameters and spacing were detected: no visible defects or structure peeling was found by scanning electron microscopy (SEM) observations, and no detectable interdiffusion between Cu and the dielectric layers was detected by energy dispersive X-ray (EDX) analyses. Electric tests indicated that the leakage currents between two adjacent TSVs were as low as 6.80 × 10-10 A when a DC voltage was ramped up from 0 to 350 V, and 2.86 × 10-9 A after a DC voltage was kept at 100 V for 200 s.

Bioinformatics Analysis Identifies IL6ST as a Potential Tumor Suppressor Gene for Triple-Negative Breast Cancer.

Improved insight into the molecular mechanisms of triple-negative breast cancer (TNBC) is required to predict prognosis and develop a new therapeutic strategy for targeted genes. The aim of this study was to identify genes significantly associated with TNBC and further analyze their prognostic significance. The Cancer Genome Atlas (TCGA) TNBC database and gene expression profiles of GSE76275 from Gene Expression Omnibus (GEO) were used to explore differentially co-expressed genes in TNBC compared with those in normal tissues and non-TNBC breast cancer tissues. Differential gene expression and weighted gene co-expression network analyses identified 24 differentially co-expressed genes. Functional annotation suggested that these genes were primarily enriched in processes such as metabolism, membrane, and protein binding. The protein-protein interaction (PPI) network further identified ten hub genes, five of which (MAPT, CBS, SOX11, IL6ST, and MEX3A) were confirmed to be differentially expressed in an independent dataset (GSE38959). Moreover, CBS and MEX3A expression was upregulated, whereas IL6ST expression was downregulated in TNBC tissues compared to that in other breast cancer subtypes. Furthermore, lower expression of IL6ST was associated with worse overall survival in patients with TNBC. Thus, IL6ST might play an important role in TNBC progression and could serve as a tumor suppressor gene for diagnosis and treatment.

Key Genes and Prognostic Analysis in HER2+ Breast Cancer.

Human epidermal growth factor 2 (HER2)+ breast cancer is considered the most dangerous type of breast cancers. Herein, we used bioinformatics methods to identify potential key genes in HER2+ breast cancer to enable its diagnosis, treatment, and prognosis prediction. Datasets of HER2+ breast cancer and normal tissue samples retrieved from Gene Expression Omnibus and The Cancer Genome Atlas databases were subjected to analysis for differentially expressed genes using R software. The identified differentially expressed genes were subjected to gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses followed by construction of protein-protein interaction networks using the STRING database to identify key genes. The genes were further validated via survival and differential gene expression analyses. We identified 97 upregulated and 106 downregulated genes that were primarily associated with processes such as mitosis, protein kinase activity, cell cycle, and the p53 signaling pathway. Visualization of the protein-protein interaction network identified 10 key genes (CCNA2, CDK1, CDC20, CCNB1, DLGAP5, AURKA, BUB1B, RRM2, TPX2, and MAD2L1), all of which were upregulated. Survival analysis using PROGgeneV2 showed that CDC20, CCNA2, DLGAP5, RRM2, and TPX2 are prognosis-related key genes in HER2+ breast cancer. A nomogram showed that high expression of RRM2, DLGAP5, and TPX2 was positively associated with the risk of death. TPX2, which has not previously been reported in HER2+ breast cancer, was associated with breast cancer development, progression, and prognosis and is therefore a potential key gene. It is hoped that this study can provide a new method for the diagnosis and treatment of HER2 + breast cancer.

An efficient method using ultrasound to accelerate aging in crabapple (Malus asiatica) vinegar produced from fresh fruit and its influencing mechanism investigation.

In this study, a kind of crabapple vinegar was developed by the method of mixed bacteria fermentation. It showed that the total acids and total esters in the vinegar increased by 30.51% and 22.67%, respectively. Simultaneously, ultrasound was used to treat the vinegar to shorten the time of aging. In addition, the HS-SPME-GC-MS results show that some volatile components had increased significantly, such as total esters, aldehydes and heterocycles. Combining OAV with radar chart of aroma active ingredients, the order of contribution to the characteristic aroma of crabapple vinegar was esters > alcohols > others > acids. Finally, ultrasonic cavitation and hydroxyl radicals were measured to further prove it could accelerate chemical reaction of crabapple vinegar. The results of FTIR showed that the hydrogen-bonded molecules had increased, while free molecules with irritating taste (such as ethanol and acetic acid) had decreased, which made the taste of crabapple vinegar softer. Results have showed that ultrasound is a promising technique for shortening aging time and it also provides the possibility to improve the taste of fruit vinegar.

Identification of key genes unique to the luminal a and basal-like breast cancer subtypes via bioinformatic analysis.

BACKGROUND: Breast cancer subtypes are statistically associated with prognosis. The search for markers of breast tumor heterogeneity and the development of precision medicine for patients are the current focuses of the field. METHODS: We used a bioinformatic approach to identify key disease-causing genes unique to the luminal A and basal-like subtypes of breast cancer. First, we retrieved gene expression data for luminal A breast cancer, basal-like breast cancer, and normal breast tissue samples from The Cancer Genome Atlas database. The differentially expressed genes unique to the 2 breast cancer subtypes were identified and subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. We constructed protein-protein interaction networks of the differentially expressed genes. Finally, we analyzed the key modules of the networks, which we combined with survival data to identify the unique cancer genes associated with each breast cancer subtype. RESULTS: We identified 1114 differentially expressed genes in luminal A breast cancer and 1042 differentially expressed genes in basal-like breast cancer, of which the subtypes shared 500. We observed 614 and 542 differentially expressed genes unique to luminal A and basal-like breast cancer, respectively. Through enrichment analyses, protein-protein interaction network analysis, and module mining, we identified 8 key differentially expressed genes unique to each subtype. Analysis of the gene expression data in the context of the survival data revealed that high expression of NMUR1 and NCAM1 in luminal A breast cancer statistically correlated with poor prognosis, whereas the low expression levels of CDC7, KIF18A, STIL, and CKS2 in basal-like breast cancer statistically correlated with poor prognosis. CONCLUSIONS: NMUR1 and NCAM1 are novel key disease-causing genes for luminal A breast cancer, and STIL is a novel key disease-causing gene for basal-like breast cancer. These genes are potential targets for clinical treatment.

Alcohol-Tolerant Platinum Electrocatalyst for Oxygen Reduction by Encapsulating Platinum Nanoparticles inside Nitrogen-Doped Carbon Nanocages.

Pt-based electrocatalysts are the most popular for direct alcohol fuel cells, but their performances easily deteriorate for the oxygen reduction reaction (ORR) at the cathode because of the alcohol crossover effect. Herein, we report the novel Pt electrocatalyst encapsulated inside nitrogen-doped carbon nanocages (Pt@NCNC), which presents excellent alcohol-tolerant ORR activity and durability in acidic media, far superior to the Pt counterpart immobilized outside the nanocages (Pt/NCNC). The superb performance is correlated with the molecule-sieving effect of the micropores penetrating through the shells of the nanocages, which admit the small-sized oxygen and ions but block the large-sized alcohols into the nanocages. This mechanism is confirmed by examining the size dependence of ORR and alcohol oxidation activities by regulating the micropores sizes. This study provides a promising strategy to develop the superior alcohol-tolerant Pt-based ORR electrocatalyst in acidic media.

Cloning the sterol carrier protein 2 genes of Japanese toad (Bufo japonicus formosus) and Chinese toad (Bufo gargarizans) and its tissue expression analysis.

In this study, to clarify the bioactive polypeptides included in the skins and secretions of Bufo, we screened the Japanese toad (Bufo japonicus formosus) skin cDNA liary by colony polymerase chain reaction (PCR), and obtained a transcript of 1 075 bp consisting of 1 37 bp 5' untranslated region (UTR), 515 bp 3' UTR and a 423 bp open reading frame (ORF) encoding a polypeptide of 140 amino acid residues (GenBank accession number: KF359945). Homolog analysis showed a 70%-96% homology with sterol carrier protein-2 (SCP-2) present in other animals, which is implicated in lipid metabolism of other organisms. The gene SCP-2 of Chinese toad (B. gargarizans) was cloned from a first strand cDNA of Bufo skin (GenBank accession number: KF381341) via PCR, whose encoding polypeptide has only one amino acid difference from that of Japanese toad. Tissue distribution analysis showed that SCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs. These findings might indicate SCP-2 being one of the active ingredients in toad skin. These findings may in turn have implications for further drug development from traditional Chinese medicine sources.