[The Effect of SIRT5 Deletion on Recovery of Hematopoietic Stem Cells after Injury in Mouse].

OBJECTIVE: To investigate the effect of deacylase Sirtuin 5 in the recovery of hematopoietic stem cells (HSCs) after treated by 5-FU in mouse. METHODS: Flow cytometry was used to analyze the effect of SIRT5 deletion on the proportion of hematopoietic stem/progenitor cells (HSPCs) in bone marrow (BM), the proportion of T cells, B cells and myeloid cells (TBM) in peripheral blood (PB) and spleen, and the development of T cells in thymus. Mouse were treated with 5-FU to study the effect of SIRT5 deletion on the cell cycle, apoptosis and the proportion of HSPCs in BM. The effect of SIRT5 deletion on the proliferation of HSCs was analyzed by flow sorting in vitro. RESULTS: SIRT5 deletion did not affect the development of T cells in thymus and the proportion of TBM cells in PB and spleen compared with wild type mice. SIRT5 deletion increased proportion of HSPCs in BM. After 5-FU treatment, the proportion of HSCs in SIRT5 deletion mice was significant decreased (P < 0.05), the HSPC in SIRT5 deletion mice was activated from G0 to G1 phase (P < 0.05), and the proportion of early apoptosis increased (P < 0.05). By monoclonal culture in vitro, the ability of HSCs to form clones in SIRT5 deletion mice was decreased significantly (P < 0.05). CONCLUSION: SIRT5 deletion lead to a decreased the ability of HSCs to clone in vitro. SIRT5 deletion is not conducive to the recovery of HSPCs injury in mice under hematopoietic stress.

Adaptive fuzzy control for tendon-sheath actuated bending-tip system with unknown friction for robotic flexible endoscope.

Introduction: The tendon-sheath actuated bending-tip system (TAB) has been widely applied to long-distance transmission scenes due to its high maneuverability, safety, and compliance, such as in exoskeleton robots, rescue robots, and surgical robots design. Due to the suitability of operation in a narrow or tortuous environment, TAB has demonstrated great application potential in the area of minimally invasive surgery. However, TAB involves highly non-linear behavior due to hysteresis, creepage, and non-linear friction existing on the tendon routing, which is an enormous challenge for accurate control. Methods: Considering the difficulties in the precise modeling of non-linearity friction, this paper proposes a novel fuzzy control scheme for the Euler-Lagrange dynamics model of TAB for achieving tracking performance and providing accurate friction compensation. Finally, the asymptotic stability of the closed-loop system is proved theoretically and the effectiveness of the controller is verified by numerical simulation carried out in MATLAB/Simulink. Results: The desired angle can be reached quickly within 3 s by adopting the proposed controller without overshoot or oscillation in Tracking Experiment, demonstrating the regulation performance of the proposed control scheme. The proposed method still achieves the desired trajectory rapidly and accurately without steady-state errors in Varying-friction Experiment. The angle errors generated by external disturbances are < 1 deg under the proposed controller, which returns to zero in 2 s in Anti-disturbance Experiment. In contrast, comparative controllers take more time to be steady and are accompanied by oscillating and residual errors in all experiments. Discussion: The proposed method is model-free control and has no strict requirement for the dynamics model and friction model. It is proved that advanced tracking performance and real-time response can be guaranteed under the presence of unknown bounded non-linear friction and time-varying non-linear dynamics.

Response of a Novel KANK1::ALK Fusion to Alectinib in an Advanced Lung Adenocarcinoma: A Case Report.

More than 90 distinct fusion partners of ALK rearrangement have been identified. Different ALK fusions may exhibit different sensitivities to ALK tyrosine kinase inhibitors. The emergence of rare fusions poses significant challenges to targeted therapies. This study aimed to investigate the response of KANK1::ALK fusion to alectinib in an advanced lung adenocarcinoma. A novel KANK1::ALK fusion was identified by next-generation sequencing (NGS) and Ventana immunohistochemistry assessments. A 73-year-old woman who had never smoked was admitted with hemoptysis in May 2020. PET/CT revealed a nodule in the left upper lobe, with bilateral pulmonary and multiple lymph node metastases. The upper lobe nodule of the left lung was diagnosed as adenocarcinoma through bronchofiberscopy biopsy, resulting in a clinical diagnosis of stage IVA (cT1c,N3,M1a). Because the biopsy tissue was insufficient for NGS analysis, a blood-based genetic analysis was performed, revealing the presence of KRAS p.Q61R mutations. The patient received carboplatin and pemetrexed with pembrolizumab as first-line therapy, followed by maintenance therapy of pembrolizumab monotherapy. Although the tumor initially showed significant shrinkage, it unfortunately progressed further after 11 months. Subsequently, the patient was given carboplatin and pemetrexed with pembrolizumab again, but the tumor progression continued. An NGS using a rebiopsy of the left upper lobe tumor suggested a KANK1::ALK fusion. Alectinib was prescribed in January 2022, and a durable partial response was observed after 18 months. ALK rearrangements were observed in the broader spectrum of lung cancers. This study provided a potential treatment option for patients with KANK1::ALK fusions. Further studies are needed to understand the function of these fusions.

High sensitivity saliva-based biosensor in detection of breast cancer biomarkers: HER2 and CA15-3.

The prevalence of breast cancer in women underscores the urgent need for innovative and efficient detection methods. This study addresses this imperative by harnessing salivary biomarkers, offering a noninvasive and accessible means of identifying breast cancer. In this study, commercially available disposable based strips similar to the commonly used glucose detection strips were utilized and functionalized to detect breast cancer with biomarkers of HER2 and CA15-3. The results demonstrated limits of detection for these two biomarkers reached as low as 1 fg/ml much lower than those of conventional enzyme-linked immunosorbent assay in the range of 1∼4 ng/ml. By employing a synchronized double-pulse method to apply 10 of 1.2 ms voltage pulses to the electrode of sensing strip and drain electrode of the transistor for amplifying the detected signal, and the detected signal was the average of 10 digital output readings corresponding to those 10 voltage pulses. The sensor sensitivities were achieved approximately 70/dec and 30/dec for HER2 and CA15-3, respectively. Moreover, the efficiency of this novel technique is underscored by its swift testing time of less than 15 ms and its minimal sample requirement of only 3 µl of saliva. The simplicity of operation and the potential for widespread public use in the future position this approach as a transformative tool in the early detection of breast cancer. This research not only provides a crucial advancement in diagnostic methodologies but also holds the promise of revolutionizing public health practices.

Apatinib added when NSCLC patients get slow progression with EGFR-TKI: A prospective, single-arm study.

BACKGROUND: Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI) acquired resistance was an inevitably events in NSCLC treatment. AIMS: Intending to overcome the acquired resistance of EGFR-TKI. MATERIALS & METHODS: A clinical trial was, we enrolled 12 patients who were slowly progressing on first-generation EGFR-TKI, and added apatinib when the patients got slow progression. RESULTS: Seven patients were included in the efficacy analysis. The median PFS2 of apatinib combined with EGFR-TKI was 8.2 months (95% CI, 7.3 m-NA), and the total PFS reached 20.9 months (95% CI, 17.3 m-NA) when plus PFS1. All the adverse events were manageable. The median PFS was significantly longer for circulating tumor DNA (ctDNA)-cleared patients (8.4 months; 95% CI, 8.2-NA) than for those ctDNA not cleared (7.1 months; 95% CI, 6.9-NA) (p = 0.0082). DISCUSSION: The addition of apatinib did improve the duration of first-generation EGFR-TKI use, and the duration was better than the first-line use of third-generation EGFR-TKI. CONCLUSION: The addition of apatinib when the patients got slow progression after initial EGFR-TKI therapy may be a good treatment option and the side effects are controllable. It is possible to monitor treatment efficacy using ctDNA.

Enhancing the Hydrophobicity and Antibacterial Properties of SiCN-Coated Surfaces with Quaternization to Address Peri-Implantitis.

Peri-implantitis is a major cause of dental implant failure. This disease is an inflammation of the tissues surrounding the implant, and, while the cause is multi-factorial, bacteria is the main culprit in initiating an inflammatory reaction. Dental implants with silicon carbonitride (SiCN) coatings have several potential advantages over traditional titanium implants, but their antibacterial efficiency has not yet been evaluated. The purpose of this study was to determine the anti-bacterial potential of SiCN by modifying the surface of SiCN-coated implants to have a positive charge on the nitrogen atoms through the quaternization of the surface atoms. The changes in surface chemistry were confirmed using contact angle measurement and XPS analysis. The modified SiCN surfaces were inoculated with Streptococcus mutans (S. mutans) and compared with a silicon control. The cultured bacterial colonies for the experimental group were 80% less than the control silicon surface. Fluorescent microscopy with live bacteria staining demonstrated significantly reduced bacterial coverage after 3 and 7 days of incubation. Scanning electron microscopy (SEM) was used to visualize the coated surfaces after bacterial inoculation, and the mechanism for the antibacterial properties of the quaternized SiCN was confirmed by observing ruptured bacteria membrane along the surface.

Thymol targeting interleukin 4 induced 1 expression reshapes the immune microenvironment to sensitize the immunotherapy in lung adenocarcinoma.

Immune checkpoint blockades are the most promising therapy in lung adenocarcinoma (LUAD). However, the response rate remains limited, underscoring the urgent need for effective sensitizers. Interleukin 4 induced 1 (IL4I1) is reported to have immunoinhibitory and tumor-promoting effects in several cancers. However, the targetable value of IL4I1 in sensitizing the immunotherapy is not clear, and there is a lack of effective small molecules that specifically target IL4I1. Here, we show that silencing IL4I1 significantly remodels the immune microenvironment via inhibiting aryl hydrocarbon receptor (AHR) signaling, thereby enhancing the efficacy of anti-PD-1 antibody in LUAD, which suggests that IL4I1 is a potential drug target for the combination immunotherapy. We then identify thymol as the first small molecule targeting IL4I1 transcription through a drug screening. Thymol inhibits the IL4I1 expression and blocks AHR signaling in LUAD cells. Thymol treatment restores the antitumor immune response and suppresses the progression of LUAD in an orthotopic mouse model. Strikingly, the combination treatment of thymol with anti-PD-1 antibody shows significant tumor regression in LUAD mice. Thus, we demonstrate that thymol is an effective small molecule to sensitize the PD-1 blockade in LUAD via targeting IL4I1, which provides a novel strategy for the immunotherapy of LUAD.

Properties of SiCN Films Relevant to Dental Implant Applications.

The application of surface coatings is a popular technique to improve the performance of materials used for medical and dental implants. Ternary silicon carbon nitride (SiCN), obtained by introducing nitrogen into SiC, has attracted significant interest due to its potential advantages. This study investigated the properties of SiCN films deposited via PECVD for dental implant coatings. Chemical composition, optical, and tribological properties were analyzed by adjusting the gas flow rates of NH3, CH4, and SiH4. The results indicated that an increase in the NH3 flow rate led to higher deposition rates, scaling from 5.7 nm/min at an NH3 flow rate of 2 sccm to 7 nm/min at an NH3 flow rate of 8 sccm. Concurrently, the formation of N-Si bonds was observed. The films with a higher nitrogen content exhibited lower refractive indices, diminishing from 2.5 to 2.3 as the NH3 flow rate increased from 2 sccm to 8 sccm. The contact angle of SiCN films had minimal differences, while the corrosion rate was dependent on the pH of the environment. These findings contribute to a better understanding of the properties and potential applications of SiCN films for use in dental implants.

Lobe-specific analysis of perioperative chemotherapy for non-small cell lung cancer patients.

OBJECTIVES: Perioperative cisplatin-based chemotherapy decreases the risk of death over surgery alone and is a standard of care. Here, we examined perioperative chemotherapy indications for stage IB-III non-small cell lung cancer (NSCLC) patients according to lobe-specific analysis. METHODS: Resectable NSCLC patients with stage IB-III who received perioperative chemotherapy with and without radiotherapy after lung resection were identified from the SEER database. Propensity score matching (PSM) analysis was performed to reduce the inherent bias of retrospective studies. The Kaplan-Meier method and log-rank tests were used to assess the differences in overall survival (OS). RESULTS: The study enrolled 23,844 patients before PSM. The perioperative chemotherapy group had better OS than the nonperioperative chemotherapy group in stage IB-III NSCLC patients before and after PSM. However, subgroup analysis according to stage demonstrated that perioperative chemotherapy did not markedly benefit patients with stage IB. Furthermore, lobar subgroup analysis did not show survival advantages in primary tumors located in either the right middle lobe in stages II and III NSCLC or the right lower lobe in stage III NSCLC. CONCLUSIONS: Lobe-specific perioperative chemotherapy is recommended in NSCLC patients. For stage IB NSCLC, right middle lobe NSCLC from stage IB-III and right lower lobe NSCLC from stage III, perioperative chemotherapy might not confer survival benefits.

Lymph Node Metastases in Surgically Resected Solitary Ground-Glass Opacities: A Two-Center Retrospective Cohort Study and Pooled Literature Analysis.

BACKGROUND: An increasing body of evidence supports the noninferiority of sublobar resection compared with lobectomy in terms of survival for patients with early-stage lung cancer with ground-glass opacities (GGOs). However, few studies have focused on the incidence of lymph node (LN) metastases in these patients. We aimed to analyze N1 and N2 lymph node involvement in patients with non-small cell lung cancer (NSCLC) with GGO components stratified with different consolidation tumor ratio (CTR). PATIENTS AND METHODS: We performed two-center studies by retrospectively reviewing a total of 864 patients with NSCLC with semisolid or pure GGO manifestation (diameter ≤ 3 cm). Clinicopathologic features and outcomes were analyzed. We also reviewed 35 studies to characterize the patient with NSCLC population with the GGO manifestation. RESULTS: In both cohorts, there was no LN involvement for pure GGO NSCLC, while solid predominant GGO exhibited a relatively high LN involvement rate. On the basis of a pooled literature analysis, the incidence of pathologic mediastinal LN was 0% and 3.8% for pure and semisolid GGOs, respectively. GGO NSCLCs with CTR ≤ 0.5 also had rare LN involvement (0.1%). CONCLUSIONS: From two cohorts and pooled literature analysis, LN involvement was not observed in patients with pure GGO, and very few patients with semisolid GGO NSCLC with CTR ≤ 0.5 had LN involvement, revealing that it may be unnecessary to perform lymphadenectomy for pure GGOs, while mediastinal lymph node sampling (MLNS) is enough for semisolid GGOs with CTR ≤ 0.5. For the patients with GGO CTR > 0.5, mediastinal lymphadenectomy (MLD) or MLNS should be considered.

RNA binding proteins are potential novel biomarkers of egg quality in yellow catfish.

BACKGROUND: Egg quality is a major concern in fish reproduction and development. An effective evaluation of egg quality prior to fertilization is helpful in improving the fertilization rate and survival rate of the larva. In this study, we aim to identify quality instructors from the combination study of fertilization rate, hatching rate, embryo malformation rate and gene expression profile. RESULTS: Eggs from 25 female fish were fertilized with sperm from the same fish. The egg quality was determined by the fertilization rates, hatching rate and embryo malformation rate and divided into three categories, low-quality (< 35%), medium-quality (35 to 75%), and high-quality (> 75%). Due to the distinct difference in fertilization, hatching and embryo malformation rate between low-quality eggs and high-quality eggs, these two groups were considered for the identification of quality markers. Then RNA-seq was performed for the originally preserved eggs from the low-quality group and high-quality group. We profiled the differentially expressed genes and identified a group of RNA-binding proteins (RBPs) as potential regulators. Gene function analysis indicated that most of these genes were enriched in RNA-regulated pathways including RNA processing. The RBPs were more related to egg quality from the PLS-DA analysis. Finally, gene expression was validated by qRT-PCR. CONCLUSIONS: We found a cluster of RBP genes including igf2bp3, zar1, elavl1, rbm25b and related regulatory factors including yy1, sirt1, anp32e, btg4 as novel biomarkers of egg quality.

Origin and chromatin remodeling of young X/Y sex chromosomes in catfish with sexual plasticity.

Assembly of a complete Y chromosome is a significant challenge in animals with an XX/XY sex-determination system. Recently, we created YY-supermale yellow catfish by crossing XY males with sex-reversed XY females, providing a valuable model for Y-chromosome assembly and evolution. Here, we assembled highly homomorphic Y and X chromosomes by sequencing genomes of the YY supermale and XX female in yellow catfish, revealing their nucleotide divergences with only less than 1% and with the same gene compositions. The sex-determining region (SDR) was identified to locate within a physical distance of 0.3 Mb by FST scanning. Strikingly, the incipient sex chromosomes were revealed to originate via autosome-autosome fusion and were characterized by a highly rearranged region with an SDR downstream of the fusion site. We found that the Y chromosome was at a very early stage of differentiation, as no clear evidence of evolutionary strata and classical structure features of recombination suppression for a rather late stage of Y-chromosome evolution were observed. Significantly, a number of sex-antagonistic mutations and the accumulation of repetitive elements were discovered in the SDR, which might be the main driver of the initial establishment of recombination suppression between young X and Y chromosomes. Moreover, distinct three-dimensional chromatin organizations of the Y and X chromosomes were identified in the YY supermales and XX females, as the X chromosome exhibited denser chromatin structure than the Y chromosome, while they respectively have significantly spatial interactions with female- and male-related genes compared with other autosomes. The chromatin configuration of the sex chromosomes as well as the nucleus spatial organization of the XX neomale were remodeled after sex reversal and similar to those in YY supermales, and a male-specific loop containing the SDR was found in the open chromatin region. Our results elucidate the origin of young sex chromosomes and the chromatin remodeling configuration in the catfish sexual plasticity.

[Preliminary Recommendations on the Timing of Lung Surgery after 
Novel Coronavirus Infection in Patients with Pulmonary Nodules and Lung Cancer].

In recent years, the corona virus disease 2019 (COVID-19) pandemic has had a huge impact on the global medical, political and economic fields. Since the beginning of the COVID-19 epidemic, our understanding of the impact of COVID-19 has grown exponentially. Recently, the COVID-19 epidemic has changed rapidly in China, and there has been controversy over how to carry out surgical operations for patients with lung neoplastic lesions. Some studies have shown that lung cancer patients undergoing surgery are more likely to experience respiratory failure and perioperative death after contracting COVID-19 than the general population, however, delays in cancer treatment are also associated with increased mortality among these patients. In particular, the novel coronavirus Omikron variant has a higher transmissibility and may escape the immunity obtained through the previous novel coronavirus infection and vaccination. In order to minimize the risk of novel coronavirus infection in surgical patients, it is necessary to develop new treatment guidelines, expert consensus and preventive measures. However, the current rapid change of the epidemic situation has led to insufficient time and evidence to develop guidelines and consensus. Therefore, thoracic surgeons need to evaluate specific patient populations at higher risk of severe complications before surgery and weigh the benefit of surgical treatment against the risk of novel coronavirus infection. We try to give some recommendations on lung surgery during the current domestic epidemic situation based on the guidelines and consensus of oncology and thoracic surgery organizations in different regions on lung surgery.
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Clinical significance of preoperative neutrophil-lymphocyte ratio and platelet-lymphocyte ratio in the prognosis of resected early-stage patients with non-small cell lung cancer: A meta-analysis.

BACKGROUND: Poor prognosis is linked to peripheral blood levels of preoperative platelet-lymphocyte ratio (PLR) and neutrophil-lymphocyte ratio (NLR) in many advanced cancers. Nevertheless, whether the correlation exists in resected early-stage cases with non-small cell lung cancer (NSCLC) stays controversial. Consequently, we performed a meta-analysis to explore the preoperative NLR and PLR's prognostic significance in early-stage patients with NSCLC undergoing curative surgery. METHODS: Relevant studies that validated the link between preoperative NLR or PLR and survival results were found via the proceeding databases: PubMed, Embase, Cochrane Library, and Web of Science. The merged 95% confidence interval (CI) and hazard ratio (HR) was employed to validate the link between the NLR or PLR's index and overall survival (OS) and disease-free survival (DFS) in resected NSCLC cases. We used sensitivity and subgroup analyses to assess the studies' heterogeneity. RESULTS: An overall of 21 studies were attributed to the meta-analysis. The findings indicated that great preoperative NLR was considerably correlated with poor DFS (HR = 1.58, 95% CI: 1.37-1.82, p < 0.001) and poor OS (HR = 1.51, 95% CI: 1.33-1.72, p < 0.001), respectively. Subgroup analyses were in line with the pooled findings. In aspect of PLR, raised PLR was indicative of inferior DFS (HR = 1.28, 95% CI: 1.04-1.58, p = 0.021) and OS (HR = 1.37, 95% CI: 1.18-1.60, p < 0.001). In the subgroup analyses between PLR and DFS, only subgroups with a sample size <300 (HR = 1.67, 95% CI: 1.15-2.43, p = 0.008) and TNM staging of mixed (I-II) (HR = 1.47, 95% CI: 1.04-2.07, p = 0.028) showed that the link between high PLR and poor DFS was significant. CONCLUSIONS: Preoperative elevated NLR and PLR may act as prognostic biomarkers in resected early-stage NSCLC cases and are therefore valuable for guiding postoperative adjuvant treatment.

High sensitivity CIP2A detection for oral cancer using a rapid transistor-based biosensor module.

Oral squamous cell carcinoma (OSCC) is one of the most common lip and oral cavity cancer types. It requires early detection via various medical technologies to improve the survival rate. While most detection techniques for OSCC require testing in a centralized lab to confirm cancer type, a point of care detection technique is preferred for on-site use and quick result readout. The modular biological sensor utilizing transistor-based technology has been leveraged for testing CIP2A, and optimal transistor gate voltage and load resistance for sensing setup was investigated. Sensitivities of 1 × 10-15 g/ml have been obtained for both detections of pure CIP2A protein and HeLa cell lysate using identical test conditions via serial dilution. The superior time-saving and high accuracy testing provides opportunities for rapid clinical diagnosis in the medical space.

High volume UV LED performance testing.

There is increasing interest in deep UV Light-Emitting Diodes (LEDs) for applications in water purification, virus inactivation, sterilization, bioagent detection, and UV curing, as well as charge management control in the Laser Interferometer Space Antenna (LISA), which will be the first gravitational wave detector in space. To fully understand the current state of commercial UV LEDs and assess their performance for use on LISA, large numbers of UV LEDs need to be tested across a range of temperatures while operating in air or in a vacuum. We describe a new hardware system designed to accommodate a high volume of UV LED performance tests and present the performance testing results from over 200 UV LEDs with wavelengths in the 250 nm range.

The Effect of Amino Sugars on the Composition and Metabolism of a Microcosm Biofilm and the Cariogenic Potential against Teeth and Dental Materials.

Amino sugars N-acetylglucosamine (GlcNAc) and glucosamine (GlcN) are abundant sources of carbon and nitrogen in the oral cavity. The aim of this study was to investigate the effects of GlcNAc metabolism on the genomics and biochemistry of a saliva-derived microbial community, and on the surface integrity of human teeth and restorative surfaces. Pooled cell-containing saliva (CCS) was used to establish a microcosm biofilm in vitro in a biofilm medium (BM) containing 5 different carbohydrates. The microbial composition of each biofilm was analyzed by 16S rRNA amplicon sequencing, and the concentrations of eight organic acids were determined for selected sugars by targeted metabolomics. Meanwhile, extracted human teeth and polished titanium and ceramic disks were submerged in BM supplemented with 1% of glucose or GlcNAc, inoculated with CCS and Streptococcus mutans UA159, and incubated for 30 days. To mimic the effects of other microbial byproducts, the specimens were immersed in 10 mM hydrogen peroxide and 10 mM ammonium hydroxide for 30 days. The surface of each specimen was evaluated by profilometry for roughness (Ra) and imaged by scanning electron microscopy. The pH of the biofilm supernatant was significantly higher for the medium containing GlcNAc (p < 0.0001), and was higher in samples containing teeth than the two restorative disks for media containing the same sugar. For both teeth and titanium specimens, the samples treated with glucose-biofilm presented higher roughness values (Ra) than those with GlcNAc-biofilm and every other group. SEM images of the teeth and titanium disks largely supported the profilometry results, with glucose-biofilm samples demonstrating the largest deviation from the reference. For ceramic disks, slightly higher Ra values were obtained for the ammonia group. These findings provide the first direct evidence to support the ability of amino sugars to significantly reduce the cariogenic potential of oral biofilms by altering their biochemistry and bacterial composition. Additionally, amino sugar metabolism appears to be less detrimental to teeth and restorative surfaces than glucose metabolism.

Effect of Silicon Carbide Coating on Osteoblast Mineralization of Anodized Titanium Surfaces.

The objective of this study was to evaluate the influence of the titanium nanotube diameter and the effect of silicon carbide (SiC) coating on the proliferation and mineralization of pre-osteoblasts on titanium nanostructured surfaces. Anodized titanium sheets with nanotube diameters of 50 and 100 nm were used. The following four groups were tested in the study: (1) non-coated 50 nm nanotubes; (2) SiC-coated 50 nm titanium nanotubes; (3) non-coated 100 nm nanotubes and (4) SiC-coated 100 nm nanotubes. The biocompatibility and cytotoxicity of pre-osteoblasts were evaluated using a CellTiter-BlueCell Viability assay after 1, 2, and 3 days. After 3 days, cells attached to the surface were observed by SEM. Pre-osteoblast mineralization was determined using Alizarin-Red staining solution after 21 days of cultivation. Data were analyzed by a Kruskal−Wallis test at a p-value of 0.05. The results evidenced biocompatibility and non-cytotoxicity of both 50 and 100 nm diameter coated and non-coated surfaces after 1, 2 and 3 days. The statistical analysis indicates a statistically significant higher cell growth at 3 days (p < 0.05). SEM images after 3 days demonstrated flattened-shaped cells without any noticeable difference in the phenotypes between different diameters or surface treatments. After 21 days of induced osteogenic differentiation, the statistical analysis indicates significantly higher osteoblast calcification on coated groups of both diameters when compared with non-coated groups (p < 0.05). Based on these results, we can conclude that the titanium nanotube diameter did not play any role on cell viability or mineralization of pre-osteoblasts on SiC-coated or non-coated titanium nanotube sheets. The SiC coating demonstrated biocompatibility and non-cytotoxicity and contributed to an increase in osteoblast mineralization on titanium nanostructured surfaces when compared to non-coated groups.

Comprehensive analyses of one-carbon metabolism related genes and their association with prognosis, tumor microenvironment, chemotherapy resistance and immunotherapy in lung adenocarcinoma.

Background: Lung adenocarcinoma (LUAD) is the most common type of lung cancer and is a global public health concern. One-carbon (1C) metabolism plays a crucial role in the occurrence and development of multiple cancer types. However, there are limited studies investigating 1C metabolism in LUAD. This study aims to evaluate the prognostic value of 1C metabolism-related genes in LUAD and to explore the potential correlation of these genes with gene methylation, the tumor microenvironment, and immunotherapy. Methods: We identified 26 1C metabolism-related genes and performed a Kaplan-Meier and Cox regression analysis to evaluate the prognostic value of these genes. Consensus clustering was further performed to determine the 1C metabolism-related gene patterns in LUAD. The clinical and molecular characteristics of subgroups were investigated based on consensus clustering. CIBERSORT and ssGSEA algorithms were used to calculate the relative infiltration levels of multiple immune cell subsets. The relationship between 1C metabolism-related genes and drug sensitivity to immunotherapy was evaluated using the CellMiner database and IMvigor210 cohort, respectively. Results: The expression levels of 23 1C metabolism-related genes were significantly different between LUAD tumor tissues and normal tissues. Seventeen of these genes were related to prognosis. Two clusters (cluster 1 and cluster 2) were identified among 497 LUAD samples based on the expression of 7 prognosis-related genes. Distinct expression patterns were observed between the two clusters. Compared to cluster 2, cluster 1 was characterized by inferior overall survival (OS) (median OS = 41 vs. 60 months, p = 0.00031), increased tumor mutation burden (15.8 vs. 7.5 mut/Mb, p < 0.001), high expression of PD-1 (p < 0.001) and PD-L1 (p < 0.001), as well as enhanced immune infiltration. 1C metabolism-related genes were positively correlated with the expression of methylation enzymes, and a lower methylation level was observed in cluster 1 (p = 0.0062). Patients in cluster 1 were resistant to chemotherapy drugs including pemetrexed, gemcitabine, paclitaxel, etoposide, oxaliplatin, and carboplatin. The specific expression pattern of 1C metabolism-related genes was correlated with a better OS in patients treated with immunotherapy (median OS: 11.2 vs. 7.8 months, p = 0.0034). Conclusion: This study highlights that 1C metabolism is correlated with the prognosis of LUAD patients and immunotherapy efficacy. Our findings provide novel insights into the role of 1C metabolism in the occurrence, development, and treatment of LUAD, and can assist in guiding immunotherapy for LUAD patients.

The Biocomplex Assembled from Antigen Peptide and Toll-like Receptor Agonist Improved the Immunity against Pancreatic Adenocarcinoma In Vivo.

Purpose: One of the biggest challenges in cancer immunotherapy is generating robust cancer-specific immunity. This work describes using a biocomplex assembled from a toll-like receptor agonist CpG oligodeoxynucleotide 1826 (CpG) and a pancreatic cancer antigen peptide mesothelin for tuning pancreatic tumor immunity. Methods: This biocomplex was assembled via electrostatic interactions and characterized in size, morphology, zeta potential, and cargo loading. The effect of biocomplex on cell viability and activation of DCs and macrophages were measured by flow cytometry. The production of cytokines (GM-CSF, TNF, and IL-6) was evaluated by using ELISA kits. The effect of biocomplex on tumor cell proliferation was also evaluated by in vivo tumor model. Result: We can modulate the surface charge of the biocomplex by simply varying the ratios of the two components. In cell models, this biocomplex did not impact cell viability in the antigen-presenting cell (i.e., dendritic cell and macrophage)-directed immunity. Moreover, this biocomplex regulated the secretion of tumor-related cytokines (i.e., GM-CSF, TNF, and IL-6) and promoted the activation of immune cell surface markers (i.e., CD80+, CD86+, and CD40+). In the mouse model, the biocomplex inhibited the tumor burden effectively and promoted the production of effector cytokines. Conclusion: The present studies showed that the biocomplex with antigen peptide and toll-like receptor agonist was able to potentiate the antitumor immunity in vivo. This study will help understanding of immunity in pancreatic cancer and developing new immune therapeutic strategies for pancreatic adenocarcinoma.