Elucidating the Influence of MPT-driven necrosis-linked LncRNAs on immunotherapy outcomes, sensitivity to chemotherapy, and mechanisms of cell death in clear cell renal carcinoma.

Background: Clear cell renal carcinoma (ccRCC) stands as the prevailing subtype among kidney cancers, making it one of the most prevalent malignancies characterized by significant mortality rates. Notably,mitochondrial permeability transition drives necrosis (MPT-Driven Necrosis) emerges as a form of cell death triggered by alterations in the intracellular microenvironment. MPT-Driven Necrosis, recognized as a distinctive type of programmed cell death. Despite the association of MPT-Driven Necrosis programmed-cell-death-related lncRNAs (MPTDNLs) with ccRCC, their precise functions within the tumor microenvironment and prognostic implications remain poorly understood. Therefore, this study aimed to develop a novel prognostic model that enhances prognostic predictions for ccRCC. Methods: Employing both univariate Cox proportional hazards and Lasso regression methodologies, this investigation distinguished genes with differential expression that are intimately linked to prognosis.Furthermore, a comprehensive prognostic risk assessment model was established using multiple Cox proportional hazards regression. Additionally, a thorough evaluation was conducted to explore the associations between the characteristics of MPTDNLs and clinicopathological features, tumor microenvironment, and chemotherapy sensitivity, thereby providing insights into their interconnectedness.The model constructed based on the signatures of MPTDNLs was verified to exhibit excellent prediction performance by Cell Culture and Transient Transfection, Transwell and other experiments. Results: By analyzing relevant studies, we identified risk scores derived from MPTDNLs as an independent prognostic determinant for ccRCC, and subsequently we developed a Nomogram prediction model that combines clinical features and associated risk assessment. Finally, the application of experimental techniques such as qRT-PCR helped to compare the expression of MPTDNLs in healthy tissues and tumor samples, as well as their role in the proliferation and migration of renal clear cell carcinoma cells. It was found that there was a significant correlation between CDK6-AS1 and ccRCC results, and CDK6-AS1 plays a key role in the proliferation and migration of ccRCC cells. Impressive predictive results were generated using marker constructs based on these MPTDNLs. Conclusions: In this research, we formulated a new prognostic framework for ccRCC, integrating mitochondrial permeability transition-induced necrosis. This model holds significant potential for enhancing prognostic predictions in ccRCC patients and establishing a foundation for optimizing therapeutic strategies.

Artificial osteochondral interface of bioactive fibrous membranes mediating calcified cartilage reconstruction.

Calcified cartilage is a mineralized osteochondral interface region between the hyaline cartilage and subchondral bone. There are few reported artificial biomaterials that could offer bioactivities for substantial reconstruction of calcified cartilage. Herein we developed new poly(L-lactide-co-caprolactone) (PLCL)-based trilayered fibrous membranes as a functional interface for calcified cartilage reconstruction and superficial cartilage restoration. The trilayered membranes were prepared by the electrospinning technique, and the fibrous morphology was maintained when the chondroitin sulfate (CS) or bioactive glass (BG) particles were introduced in the upper or bottom layer, respectively. Although 30% BG in the bottom layer led to a significant decrease in tensile resistance, the inorganic ion release was remarkably higher than that in the counterpart with 10% BG. The in vivo studies showed that the fibrous membranes as osteochondral interfaces exhibited different biological performances on superficial cartilage restoration and calcified cartilage reconstruction. All of the implanted host hyaline cartilage enabled a self-healing process and an increase in the BG content in the membranes was desirable for promoting the repair of the calcified cartilage with time. The histological staining confirmed the osteochondral interface in the 30% BG bottom membrane maintained appreciable calcified cartilage repair after 12 weeks. These findings demonstrated that such an integrated artificial osteochondral interface containing appropriate bioactive ions are potentially applicable for osteochondral interface tissue engineering.

Rational design of nonstoichiometric bioceramic scaffolds via digital light processing: tuning chemical composition and pore geometry evaluation.

Bioactive ceramics are promising candidates as 3D porous substrates for bone repair in bone regenerative medicine. However, they are often inefficient in clinical applications due to mismatching mechanical properties and compromised biological performances. Herein, the additional Sr dopant is hypothesized to readily adjust the mechanical and biodegradable properties of the dilute Mg-doped wollastonite bioceramic scaffolds with different pore geometries (cylindrical-, cubic-, gyroid-) by ceramic stereolithography. The results indicate that the compressive strength of Mg/Sr co-doped bioceramic scaffolds could be tuned simultaneously by the Sr dopant and pore geometry. The cylindrical-pore scaffolds exhibit strength decay with increasing Sr content, whereas the gyroid-pore scaffolds show increasing strength and Young's modulus as the Sr concentration is increased from 0 to 5%. The ion release could also be adjusted by pore geometry in Tris buffer, and the high Sr content may trigger a faster scaffold bio-dissolution. These results demonstrate that the mechanical strengths of the bioceramic scaffolds can be controlled from the point at which their porous structures are designed. Moreover, scaffold bio-dissolution can be tuned by pore geometry and doping foreign ions. It is reasonable to consider the nonstoichiometric bioceramic scaffolds are promising for bone regeneration, especially when dealing with pathological bone defects.

Stroke Detector and Structure Based Models for Character Recognition: A Comparative Study.

Characters, which are man-made symbols composed of strokes arranged in a certain structure, could provide semantic information and play an indispensable role in our daily life. In this paper, we try to make use of the intrinsic characteristics of characters and explore the stroke and structure-based methods for character recognition. First, we introduce two existing part-based models to recognize characters by detecting the elastic strokelike parts. In order to utilize strokes of various scales, we propose to learn the discriminative multi-scale stroke detector-based representation (DMSDR) for characters. However, the part-based models and DMSDR need to manually label the parts or key points for training. In order to learn the discriminative stroke detectors automatically, we further propose the discriminative spatiality embedded dictionary learning-based representation (DSEDR) for character recognition. We make a comparative study of the performance of the tree-structured model (TSM), mixtures-of-parts TSM, DMSDR, and DSEDR for character recognition on three challenging scene character recognition (SCR) data sets as well as two handwritten digits recognition data sets. A series of experiments is done on these data sets with various experimental setup. The experimental results demonstrate the suitability of stroke detector-based models for recognizing characters with deformations and distortions, especially in the case of limited training samples.

The effect of alendronate on the expression of important cell factors in osteoclasts.

This study investigated the effects of alendronate (ALN) on critical cell factors in osteoclasts. RAW 264.7 cells were induced by sRANKL to change to mature osteoclasts. On the sixth day of incubation, the osteoclasts were treated with ALN at various concentrations and for different incubation times. The concentration groups included 10-5 M, 10-6 M and 10-7 M ALN, respectively. The cells were incubated for 0 (control group), 2, 4, 6 and 8 h for each dose group. The mRNA and protein expression of tartrate­resistant acid phosphatase, carbonic anhydrase II, osteoclast­associated receptor and FAS/FASL genes in osteoclasts was analyzed. A concentration- and time­dependent decrease in the mRNA and protein expression levels of the five genes was observed, and no significant difference between the two control groups was observed (P>0.05). Notably, significant differences between any two experimental groups were observed (P<0.05). Thus, ALN significantly decreased the expression of critical factors involved in osteoclast function.

[Construction and analysis of one kind of chitosan-coated BMP-2 nanoparticles as genetic carrier].

OBJECTIVE: To construct an excellent kind of chitosan-coated BMP-2 nanoparticles as genetic carrier. METHODS: Chitosan-coated BMP-2 nanoparticles as genetic carrier, including C-B50 group, C-B100 group and C-B200 group, were constructed through re-coacervation and gene recombination methods. This carrier was studied by using morphological observation, enveloping test, anti- DNase I digested test and MTT assay. RESULTS: One kind of excellent chitosan-coated BMP-2 nanoparticles as genetic carrier was constructed. Particle diameter was about (90 +/- 20) nm. Enveloping ratio of C-B50 group, C-B100 group and C-B200 group was 76.31% + 1.58%, 84.49% + 1.81% and 81.69% + 1.77%, respectively. Genetic carrier could prevent objective gene from DNase I digestion. The growth inhibiting ratio to bone marrow stem cells was increased as the increase of genetic carrier dose and contact time (P < 0.05), but maximal dose of carrier had no obvious toxic effect on cells growth in vitro. CONCLUSION: The constructed chitosan-coated BMP-2 nanoparticle material was a promising genetic carrier in bone tissue engineering.

[Effects of fluid shear stress strength on mRNA expression of ATP6V1a1 in polarized osteoclasts].

OBJECTIVE: To observe effects of fluid shear stress strength on mRNA expression of ATP6V1a1 in rat polarized osteoclasts. METHODS: Rat polarized osteoclasts suffered 0.0 (control group), 0.9, 2.9, 8.7 and 26.3 dynes/cm2 fluid shear stress for 30 min. mRNA expression of ATP6V1a1 was detected by Real-Time fluorescent quantitation PCR. RESULTS: mRNA expression of ATP6V1a1 in 0.0 (control group), 0.9, 2.9, 8.7 and 26.3 dynes/cm2 groups was (1.14 +/- 0.06) x 10(6), (1.62 +/- 0.09) x 10(6), (2.28 +/- 0.13) x 10(6), (3.24 +/- 0.18) x 10(6), (9.16 +/- 0.53) x 10(6) copy numbers, respectively (P < 0.05). CONCLUSION: In the present study, polarized osteoclasts are sensitive to fluid shear stress. mRNA expression of ATP6V1a1 has increscent tendency along with increasing of fluid shear stress strength.

Effects of fluid shear stress on mRNA expression of carbonic anhydrase II in polarized rat osteoclasts.

The present study was designed to determine the effects of fluid shear stress on the mRNA expression of carbonic anhydrase II (CAII) in polarized rat osteoclasts. Cellular morphology of the polarized osteoclasts generated by a mechanical anatomical technique was examined by tartrate-resistant acid phosphatase (TRAP) staining and the osteoclastic resorption of dentine slices. The polarized osteoclasts were then stress-loaded by using a flow shear stress device newly developed by the osteoclast research group (patent number 200420034438; China), at 9 dyne/cm(2) for various time periods [0 (control group), 15, 30, 60, and 120 min], or at various stress levels [0 (control), 0.9, 2.9, 8.7, and 26.3 dyne/cm(2)] for 30 min. The mRNA expression of CAII was quantified using real-time fluorescent quantitative PCR (RT-PCR) and the data were analyzed with SPSS 12.0 software. The polarized osteoclasts were larger than regular monocytes (about 30 microm diameter) with irregular configuration, and the majority of polarized osteoclasts appeared to be spherical and had approximately 2-20 nuclei. The TRAP positive polarized osteoclasts showed asymmetrical red staining in the cytoplasm, and had many filaments and vacuoles. These cells formed resorptive pits in dentine slices. The levels of CAII mRNA expression were shown to be time-dependent, with the E+5 copy numbers being 7.88+/-0.09, 11.14+/-0.12, 15.83+/-0.18, 1.94+/-0.02, and 1.37+/-0.01 in cells treated at 9 dyne/cm(2) for 0, 15, 30, 60 and 120 min, respectively (P < 0.05). The levels of CAII mRNA expression (E+5 copy numbers) in cells treated with the stress levels of 0, 0.9, 2.9, 8.7 and 26.3 dyne/cm(2) were 7.97+/-0.201, 11.26+/-0.688, 15.94+/-0.201, 31.88+/-1.496, and 45.08+/-2.639, respectively (P < 0.05). These results indicate that there is a relationship between the fluid shear stress and the mRNA expression of CAII in polarized rat osteoclasts.