Evaluating the accuracy of a new robotically assisted system in cadaveric total knee arthroplasty procedures.

BACKGROUND: Robot-assisted total knee arthroplasty (TKA) has been shown to facilitate high-precision bone resection, which is an important goal in TKA. The aim of this cadaveric study was to analyze the accuracy of the target angle and bone resection thickness of a recently introduced robotic TKA system. METHODS: This study used 4 frozen cadaveric specimens (8 knees), 2 different implant designs, navigation, and a robotic system. The 4 surgeons who participated in this study were trained and familiar with the basic principles and operating procedures of this system. The angle of the bone cuts performed using the robotic system was compared with the target angles from the intraoperative plan. For each bone cut, the resection thickness was recorded and compared with the planned resection thickness. RESULTS: The mean angular difference for all specimens was less than 1°, and the standard deviation was less than 2°. The mean difference between the planned and measured angles was close to 0 and not significantly different from 0 except for the difference in the frontal tibial component angle, which was 0.88°. The mean difference in the hip-knee-ankle axis angle was - 0.21°± 1.06°. The mean bone resection difference for all specimens was less than 1 mm, and the standard deviation was less than 0.5 mm. CONCLUSIONS: The results of the cadaveric experimental study showed that the new TKA system can realize highly accurate bone cuts and achieve planned angles and resection thicknesses. Despite the limitations of small sample sizes and large differences between cadaveric and clinical patients, the accuracy of cadaveric experiments provides strong support for subsequent clinical trials.

Study on the Application Effect of Three-dimensional Reconstruction Technology in Locating Small Pulmonary Nodules During VATS Surgery.

Objective: To compare the positioning effect of three-dimensional reconstruction technology and Hook-wire puncture operation on small pulmonary nodules during video-assisted thoracoscopic surgery (VATS), and evaluate its effectiveness, efficiency, and safety. Methods: The subjects of this study were 50 patients with small pulmonary nodules admitted to the Department of Cardiothoracic Surgery of Heilongjiang Provincial Hospital from January 2020 to December 2022, and all underwent thoracoscopic surgical resection. All study subjects met the inclusion criteria, grouping according to the intraoperative positioning method, the control group (n = 25) used Hook-wire puncture positioning, and the observation group (n = 25) used three-dimensional reconstruction technology. The positioning effect, pain level, and postoperative complications were compared between the two groups. Results: The incidence rate of complications after puncture was 16.00% in the control group and 4.00% in the observation group, the complication rate in the observation group was significantly lower; the positioning success rate of the observation group was 96.00%, which was higher than that of the control group (92.00%). The operation time (32.25±6.08) min was lower than (38.50±7.12) min in the control group. The two groups had no statistical significance in the wedge resection success rate, VAS score, and complication rate (P > .05). Conclusion: Three-dimensional reconstruction technology mainly makes preliminary judgments on the location, shape, size, and relationship between nodules and surrounding tissues based on preoperative CT scan images. It can select suitable scanning locations, map puncture paths, and anchor them in and around small lung nodules. The operation is simple, and the positioning success rate is high. The existence of three-dimensional reconstruction technology to position the guide wire can quickly shorten the time to detect lesions, shorten the time of VATS, reduce the occurrence of pulmonary infection in patients, and improve the prognosis.

In Silico Design and Synthesis of Antifungal Peptides Guided by Quantitative Antifungal Activity.

Antifungal peptides (AFPs) are emerging as promising candidates for advanced antifungal therapies because of their broad-spectrum efficacy and reduced resistance development. In silico design of AFPs, however, remains challenging, due to the lack of an efficient and well-validated quantitative assessment of antifungal activity. This study introduced an AFP design approach that leverages an innovative quantitative metric, named the antifungal index (AFI), through a three-step process, i.e., segmentation, single-point mutation, and global multipoint optimization. An exhaustive search of 100 putative AFP sequences indicated that random modifications without guidance only have a 5.97-20.24% chance of enhancing antifungal activity. Analysis of the search results revealed that (1) N-terminus truncation is more effective in enhancing antifungal activity than the modifications at the C-terminus or both ends, (2) introducing the amino acids within the 10-60% sequence region that enhance aromaticity and hydrophobicity are more effective in increasing antifungal efficacy, and (3) incorporating alanine, cysteine, and phenylalanine during multiple point mutations has a synergistic effect on enhancing antifungal activity. Subsequently, 28 designed peptides were synthesized and tested against four typical fungal strains. The success rate for developing promising AFPs, with a minimal inhibitory concentration of ≤5.00 µM, was an impressive 82.14%. The predictive and design tool is accessible at https://antifungipept.chemoinfolab.com.

Is AI 3D-printed PSI an accurate option for patients with developmental dysplasia of the hip undergoing THA?

BACKGROUND: In traditional surgical procedures, significant discrepancies are often observed between the pre-planned templated implant sizes and the actual sizes used, particularly in patients with congenital hip dysplasia. These discrepancies arise not only in preoperative planning but also in the precision of implant placement, especially concerning the acetabular component. Our study aims to enhance the accuracy of implant placement during Total Hip Arthroplasty (THA) by integrating AI-enhanced preoperative planning with Patient-Specific Instrumentation (PSI). We also seek to assess the accuracy and clinical outcomes of the AI-PSI (AIPSI) group in comparison to a manual control group. METHODS: This study included 60 patients diagnosed with congenital hip dysplasia, randomly assigned to either the AIPSI or manual group, with 30 patients in each. No significant demographic differences between were noted the two groups. A direct anterior surgical approach was employed. Postoperative assessments included X-rays and CT scans to measure parameters such as the acetabular cup anteversion angle, acetabular cup inclination angle, femoral stem anteversion angle, femoral offset, and leg length discrepancy. Functional scores were recorded at 3 days, 1 week, 4 weeks, and 12 weeks post-surgery. Data analysis was conducted using SPSS version 22.0, with the significance level was set at α = 0.05. RESULTS AND CONCLUSION: The AIPSI group demonstrated greater prosthesis placement accuracy. With the aid of PSI, AI-planned THA surgery provides surgeons with enhanced precision in prosthesis positioning. This approach potentially offers greater insights and guidelines for managing more complex anatomical variations or cases.

The Function of Different Subunits of the Molecular Chaperone CCT in the Microsporidium Nosema bombycis: NbCCTζ Interacts with NbCCTα.

Chaperonin containing tailless complex polypeptide 1 (CCT) is a molecular chaperone protein that consists of eight completely different subunits and assists in the folding of newly synthesized peptides. The zeta subunit of CCT is a regulatory factor for the folding and assembly of cytoskeletal proteins as individuals or complexes. In this study, the zeta subunit of Nosema bombycis (NbCCTζ) is identified for the first time. The complete ORF of the NbCCTζ gene is 1533 bp in length and encodes a 510 amino acid polypeptide. IFA results indicate that NbCCTζ is colocalized with actin and ß-tubulin in the cytoplasm during the proliferative phase and that NbCCTζ is completely colocalized with NbCCTα in the cytoplasm of N. bombycis throughout the entire life cycle. Furthermore, the yeast two-hybrid assay revealed that the NbCCTζ interacts with NbCCTα. The transcriptional level of NbCCTζ is significantly downregulated by knocking down the NbCCTα gene, while the transcriptional level of NbCCTα is downregulated after knocking down the NbCCTζ gene. These results suggest that NbCCTζ may play a vital role in the proliferation of N. bombycis by coordinating with NbCCTα.

Clinical application of artificial intelligence-assisted three-dimensional planning in direct anterior approach hip arthroplasty.

PURPOSE: The objective of this study was to investigate the efficacy of an artificial intelligence-assisted 3D planning system (AIHIP) in total hip arthroplasty by direct anterior approach and assess the reliability of the AIHIP preoperative program in terms of both interobserver and intraobserver agreement. METHODS: A retrospective analysis was conducted on patients who underwent unilateral primary THA via direct anterior approach from June 2019 to March 2022. Participants were randomly assigned to receive either the AIHIP system (n = 220) or the 2D template (control group) (n = 220) for preoperative planning. The primary outcome aimed to evaluate the correspondence between the prosthesis selected intro-operation and the one planned preoperatively, as well as to calculate the intraclass correlation coefficient (ICC). Secondary outcomes included operation time, intraoperative blood loss, fluoroscopy times, Harris hip score (HHS), lower limb length difference (LLD), femoral offset (FO), and bilateral femoral offset difference. RESULTS: No significant differences were observed in gender, age, body mass index (BMI), aetiology, and American Society of Anesthesiologists (ASA) score between the two groups. Both planning methods exhibited good intraobserver agreement for component planning (ICC: 0.941-0.976). Interobserver agreement for component planning was comparable between the two methods (ICC: 0.882-0.929). In the AIHIP group, the accuracy of acetabular cup and femoral stem prosthetics planning significantly improved, with accuracies within the size range of ± 0 and ± 1 being 76.8% and 90.5% and 79.5% and 95.5%, respectively. All differences between two groups were statistically significant (p < 0.05). Patients receiving AIHIP preoperative planning experienced shorter operation times, reduced intraoperative blood loss, fewer fluoroscopy times, and lower leg length discrepancy (LLD) (p < 0.05). Moreover, they demonstrated a higher Harris hip score (HHS) at three days post-surgery (p < 0.05). However, no significant differences were found in femoral offset (FO), difference of bilateral femoral offsets, and HHS at 1 month after the operation. CONCLUSION: Utilizing AIHIP for preoperative planning of direct anterior approach THA can significantly enhance the accuracy of prosthetic sizing with good reliability, decrease operation time, reduce intraoperative blood loss, and more effectively restore the length of both lower limbs. This approach has greater clinical application value.

Cuprotosis clusters predict prognosis and immunotherapy response in low-grade glioma.

Cuprotosis, an emerging mode of cell death, has recently caught the attention of researchers worldwide. However, its impact on low-grade glioma (LGG) patients has not been fully explored. To gain a deeper insight into the relationship between cuprotosis and LGG patients' prognosis, we conducted this study in which LGG patients were divided into two clusters based on the expression of 18 cuprotosis-related genes. We found that LGG patients in cluster A had better prognosis than those in cluster B. The two clusters also differed in terms of immune cell infiltration and biological functions. Moreover, we identified differentially expressed genes (DEGs) between the two clusters and developed a cuprotosis-related prognostic signature through the least absolute shrinkage and selection operator (LASSO) analysis in the TCGA training cohort. This signature divided LGG patients into high- and low-risk groups, with the high-risk group having significantly shorter overall survival (OS) time than the low-risk group. Its predictive reliability for prognosis in LGG patients was confirmed by the TCGA internal validation cohort, CGGA325 cohort and CGGA693 cohort. Additionally, a nomogram was used to predict the 1-, 3-, and 5-year OS rates of each patient. The analysis of immune checkpoints and tumor mutation burden (TMB) has revealed that individuals belonging to high-risk groups have a greater chance of benefiting from immunotherapy. Functional experiments confirmed that interfering with the signature gene TNFRSF11B inhibited LGG cell proliferation and migration. Overall, this study shed light on the importance of cuprotosis in LGG patient prognosis. The cuprotosis-related prognostic signature is a reliable predictor for patient outcomes and immunotherapeutic response and can help to develop new therapies for LGG.

Impact of preoperative chemotherapy on cutaneous wound healing in lung cancer patients: A meta-analysis.

As part of their treatment, lung cancer patients frequently endure thoracic oncological surgery, with preoperative chemotherapeutic interventions being the common approach. However, the potential impact of these chemotherapeutic regimens on cutaneous wound healing outcomes following surgery remains the topic of considerable clinical interest. This meta-analysis sought to evaluate comprehensively the effect of preoperative chemotherapeutic regimens on cutaneous wound healing in lung cancer patients following thoracic oncological surgery. Extensive literature searches were conducted using the leading databases PubMed, Embase, Cochrane Library and Scopus. Eight studies out of 1342 identified satisfied the inclusion criteria. Consideration was given to both randomized controlled trials (RCTs) and observational studies. Data pertaining to study characteristics, patient demographics, chemotherapeutic regimens and wound healing outcomes were extracted with great attention to detail. The examination of these varied studies provided insights into the fluctuations in rates of recovery following treatment, incidences of wound infections and frequencies of surgical complications. The research studies provided odds ratios for recovery that varied significantly in magnitude from 0.95 to 0.38, with regard to the probability of wound infection. Furthermore, a range of odds ratios for complications were disclosed, with certain odds ratios displaying narrow confidence intervals. The complexity of the effect of preoperative chemotherapy on wound closure subsequent to thoracic oncologic surgery is highlighted by our findings. The results underscore the need for individualized treatment strategies for lung cancer patients undergoing surgical procedures that strike a balance between patient safety and optimal clinical outcomes.

Camera-based Respiratory Imaging for Thoracic Asymmetry in Thoracic Surgery Patients.

The current tool for assessing thoracic asymmetry of thoracic surgery patients is inappropriate for timely or frequent clinical routines due to its dependency on empirical physical examinations or specialized machines. This study investigates the camera-based respiratory imaging for screening thoracic asymmetry, in an intelligent and convenient way. The respiratory heatmaps are generated based on the respiratory magnitudes, phases and angles extracted from the chest video, and bilateral chest region of interest are compared statistically. Due to the variability of chest respiratory direction, spatial enhancement (SDR and SPCA) algorithms are proposed to magnify the respiratory energy. The proposed framework was validated in a clinical trial involving 31 patients, recorded by a smartphone camera. A high correlation was found between the camera measurements and patients' thoracic status in both the visual imaging and quantified indices. The respiratory imaging of camera shows a clear potential for assessing chest abnormalities of thoracic surgery patients.

Utilizing the visual analogue scale (VAS) to monitor and manage pain in post-operative skin wounds after thoracic surgery.

Due to the global increase in thoracic interventions, there is greater emphasis on refining post-operative care. The purpose of this study was to validate the visual analogue scale (VAS) as the valid method for measuring post-operative pain in thoracic surgery patients. From January 2020 to June 2022, this cross-sectional study investigated 240 adult patients who underwent elective thoracic surgeries in Thoracic Surgery Department of Heilongjiang Provincial Hospital. The participants were instructed to rate their discomfort using VAS at predetermined intervals after surgery. The following demographic and clinical information was recorded: age, gender, type of thoracic surgery, and history of chronic pain. Results showed a progressive decline in post-operative VAS scores over 72 h: 8.2 immediately after surgery, 6.0 at 24 h, 5.4 at 48 h, and 3.6 by 72 h. There were notable correlations between VAS scores and chronic pain history, with moderately positive correlation of 0.40 being observed. Mean scores for males and females were 3.8 and 3.9, respectively. The analysis by age revealed comparable mean scores for age categories below and above 40. With the exception of thoracic wall resection, which resulted in an average VAS score of 4.1 ± 1.0 (p < 0.05), the type of surgery had the minimal effect on variability of pain scores. The VAS is a reliable method for evaluating post-thoracic surgery discomfort. Given the substantial impact of pain history on VAS scores, there is an urgent need for personalized pain management strategies to improve post-operative care.

Ssn6 Interacts with Polar Tube Protein 2 and Transcriptional Repressor for RNA Polymerase II: Insight into Its Involvement in the Biological Process of Microsporidium Nosema bombycis.

Nosema bombycis is a representative species of Microsporidia, and is the pathogen that causes pebrine disease in silkworms. In the process of infection, the polar tube of N. bombycis is injected into the host cells. During proliferation, N. bombycis recruits the mitochondria of host cells. The general transcriptional corepressor Ssn6 contains six tetratricopeptide repeats (TPR) and undertakes various important functions. In this study, we isolated and characterized Nbssn6 of the microsporidium N. bombycis. The Nbssn6 gene contains a complete ORF of 1182 bp in length that encodes a 393 amino acid polypeptide. Indirect immunofluorescence assay showed that the Ssn6 protein was mainly distributed in the cytoplasm and nucleus at the proliferative phase of N. bombycis. We revealed the interaction of Nbssn6 with polar tube protein 2 (Nbptp2) and the transcriptional repressor for RNA polymerase II (Nbtrrp2) by Co-IP and yeast two-hybrid assays. Results from RNA interference further confirmed that the transcriptional level of Nbptp2 and Nbtrrp2 was regulated by Nbssn6. These results suggest that Nbssn6 impacts the infection and proliferation of N. bombycis via interacting with the polar tube protein and transcriptional repressor for RNA polymerase II.

Two New Alkaloids from Roots of Zea mays and Their Cytotoxic Activity against Hep3B and SW480 Cells.

Two undescribed alkaloids, along with seven known compounds, were isolated from the roots of Zea mays (RM). Their chemical structures were elucidated based on extensive analyses of HR-ESI-MS, 1D and 2D NMR, and CD spectra. Two new alkaloids exhibited moderate inhibition of Hep3B (IC50 values of 11.7±2.4 and 14.2±3.6 µM) and SW480 cells (IC50 values of 33.4±8.2 and 47.3±5.8 µM) compared to that of the positive control compound, Oxaliplatin, IC50 value of 8.4±1.7 and 45.8±5.6 µM, respectively.

The mechanism of trans-δ-viniferin inhibiting the proliferation of lung cancer cells A549 by targeting the mitochondria.

Trans-δ-viniferin (TVN), as a natural extract, is a resveratrol dimer with attractive biological activities, particularly its anti-tumor character. However, the mechanism of TVN interfering with cancerous proliferation has not been fully understood. Herein in this study, we found that TVN could trigger cancerous mitochondrial membrane potential (ΔΨm) reduction, with intracellular reactive oxidative species (ROS) level increasing, leading to apoptosis, which makes TVN a promising candidate for lung cancer cells A549 treatment. Therefore, this study provides TVN as an option to meet the demand for higher antitumor availability with lower biotoxicity and other clinical applications.

Graphene Quantum Dots Modulate Stress Granule Assembly and Prevent Abnormal Phase Transition of Fused in Sarcoma Protein.

Protein liquid-liquid phase separation (LLPS) plays a crucial role in mediating dynamic assembly of different membraneless organelles such as stress granules (SGs). Dysregulation of dynamic protein LLPS leads to aberrant phase transition and amyloid aggregation which is closely associated with neurodegenerative diseases. In this study, we found that three types of graphene quantum dots (GQDs) exhibit potent activity in preventing SG formation and promoting SG disassembly. We next demonstrate that GQDs can directly interact with the SGs-containing protein fused in sarcoma (FUS), inhibit and reverse FUS LLPS, and prevent its abnormal phase transition. Moreover, GQDs display superior activity in preventing amyloid aggregation of FUS and disaggregating preformed FUS fibrils. Mechanistic study further demonstrates that GQDs with different edge-site exhibit distinct binding affinity to FUS monomers and fibrils, which accounts for their distinct activities in modulating FUS LLPS and fibrillation. Our work reveals the potent capability of GQDs in modulating SG assembly, protein LLPS, and fibrillation and sheds light on rational design of GQDs as effective modulators of protein LLPS for therapeutics application.

Extra-articular Blocking Technique to Resolve Severe Acetabular Bone Defect in Developmental Dysplasia of the Hip.

OBJECTIVE: The reconstruction of acetabular bone defect in developmental dysplasia of the hip (DDH) is a great challenge. Although several successful solutions have been raised, their efficacy and reliability have not been fully substantiated. This work aims to present a simple, economic and effective acetabular reconstructive technique to resolve the massive acetabular bone defect in DDH scenario. METHODS: This is a case series and observational study investigating the effectiveness and safety of extra-articular blocking technique in patients diagnosed as DDH of Crowe type II-III and Hartofilakidis B. Sixteen consecutive patients indicated for extra-articular blocking and treated with total hip arthroplasty were enrolled in this series from January 2019 to August 2020. The outcome measures included the surgical indicators such as acetabular coverage, prosthesis position, operational time, medical cost, and short-term follow-up indicators such as complications profile, patient-reported functional scales, overall recovery after surgery, and radiographic bone integration and remodeling. Their medical documentation and follow-up records were carefully reviewed with ethical approval. RESULTS: The mean values of postoperative acetabular component inclination and anteversion were 42.3° ± 2.1° and 16.4° ± 1.8°, with an average acetabular coverage of 92.1%. The mean cost reduction for patients treated with this technique compared with those treated with trabecular metal augmentation was 15.3%. The mean time until walking under full-weight bearing decreased by 3.5 weeks compared with patients treated with autologous bone grafting. Within an average observational period of 18 months, the mean improvements in Harris hip score and WOMAC score were 31 and 22 points, respectively, which were identical to those with bone graft and metal augmentation techniques. No complications such as dislocation, acetabular loosening, periprosthetic joint infection, and limb length discrepancy were recorded. No signs of translucent line formation, third-party reaction, and wear-associated osteolysis were identified. CONCLUSION: The extra-articular blocking can work simply and effectively to address acetabular bone defect in DDH patients of Crowe II-III and Hartofilakidis B, as evidenced by cost-effectiveness and instant weight-bearing advantages, low failure rate, and early osteointegration and remodeling.

Acetabular Diameter Assessment and Three-Dimensional Simulation for Acetabular Reconstruction in Dysplastic Hips.

BACKGROUND: This study aimed to investigate the relationship between acetabular width, three-dimensional (3D) simulation, and surgical results in total hip arthroplasty patients who have developmental dysplasia of the hip (DDH). METHODS: This retrospective study included 216 DDH cases. Inner and outer acetabular width (OAW) was measured at the plane passing through the center of acetabular fossa. 3D simulation and 2D standard templating were performed. The actual cup size and the use of augments during surgery were recorded. Association among the indices and their distribution in different types of DDH were analyzed. RESULTS: A difference of 13 to 14 millimeters (mm) was found between the inner acetabular width and actual cup size used in type II, III, and IV cases, while the difference was 0.2 to 3.6 mm for OAW. The accuracy of 2D templating and 3D simulation in predicting cup size was comparable in Crowe type I (86.5 versus 76%, P = .075), type II (72.7 versus 51.5%, P = .127), and type III (93.3 versus 66.7%, P = .169). The 3D simulation was significantly more accurate in Crowe type IV (89.1% versus 60.9%, P = .001). Augments and bone grafts were significantly more commonly used in type II (25%) than in the other types (0 to 6.5%). CONCLUSION: OAW more accurately predicted actual cup size than inner acetabular width. The supero-lateral acetabular bone defects in type II cases require additional attention. Compared with 2D templating, 3D simulation is more accurate in predicting actual cup size in dysplastic hips with severe deformity and may be recommended in these selected cases, especially for Crowe IV patients.

Development and validation of an inflammatory response-related prognostic model and immune infiltration analysis in glioblastoma.

Background: Despite receiving standard treatment, the prognosis of glioblastoma (GBM) patients is still poor. Considering the heterogeneity of each patient, it is imperative to identify reliable risk model that can effectively predict the prognosis of each GBM patient to guide the personalized treatment. Methods: Transcriptomic gene expression profiles and corresponding clinical data of GBM patients were downloaded from The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) databases. Inflammatory response-related genes were extracted from Gene Set Enrichment Analysis (GSEA) website. Univariate Cox regression analysis was used for prognosis-related inflammatory genes (P<0.05). A polygenic prognostic risk model was constructed using least absolute shrinkage and selection operator (LASSO) Cox regression analysis. Validation was performed through CGGA cohort. Overall survival (OS) was compared by Kaplan-Meier analysis. A nomogram was plotted to accurately predict the prognosis for each patient. GSEA was used for the pathway enrichment analysis. The single sample GSEA (ssGSEA) algorithm was implemented to conduct the immune infiltration analysis. The potential role of oncostatin M receptor (OSMR) in GBM was investigated through the in vitro experiment. Results: A prognostic risk model consisting of 4 genes (PTPRN, OSMR, MYD88, and EFEMP2) was developed. GBM patients in the high-risk group had worse OS. The time-dependent ROC curves showed an area under the curve (AUC) of 0.782, 0.765, and 0.784 for 1-, 2-, and 3-year survival in TCGA cohort, while the AUC in the CGGA cohort was 0.589, 0.684, and 0.785 at 1, 2, and 3 years, respectively. The risk score, primary-recurrent-secondary (PRS) type, and isocitrate dehydrogenase (IDH) mutation could predict the prognosis of GBM patients well. The nomogram accurately predicted the 1-, 2-, and 3-year OS for each patient. Immune cell infiltration was associated with the risk score and the model could predict immunotherapy responsiveness. The expression of the prognostic gene was correlated with the sensitivity to antitumor drugs. Interference of OSMR inhibited proliferation and migration and promoted apoptosis of GBM cells. Conclusions: The prognostic model based on 4 inflammatory response-related genes had reliable predictive power to effectively predict clinical outcome in GBM patients and provided the guide for the personalized treatment.

Identification and subcellular colocalization of protein transport protein Sec61α and Sec61γ in Nosema bombycis.

The main function of Sec61 complex is participating in the transport of polypeptide chains across the endoplasmic reticulum. The Sec61α subunit is the largest subunit of the Sec61 complex and shows high degree of conservation. In this study, we identified the NbSec61α and NbSec61γ genes in the microsporidian Nosema bombycis for the first time. Multiple sequence alignment showed that the sequence similarity between NbSec61α and homologous proteins of other microsporidia was greater than 48 %. NbSec61α contains a "plug" domain (amino acids 40-74) unique to the Sec61/SecY complex. Phylogenetic analysis based on NbSec61α and NbSec61γ indicated that the N. bombycis was closely related to Nosema granulosis, Nosema ceranae and Nosema apis. Indirect immunfluorescence assay showed that NbSec61α and NbSec61γ were mainly distributed in the perinuclear region of N. bombycis in different developmental phases. qRT-PCR results revealed that the expression level of NbSec61α gene increased in the early stage and reached the highest at 48 h, then decreased in the late stages. After knockdown of NbSec61α, the expression of NbSec61α, NbSec61γ and NbssrRNA genes were all significantly down-regulated. These results suggest that the NbSec61α and NbSec61γ may play an important role in the intracellular development of N. bombycis.

Prognostic Role of M6A-Associated Immune Genes and Cluster-Related Tumor Microenvironment Analysis: A Multi-Omics Practice in Stomach Adenocarcinoma.

N6-methylandrostenedione (m6A) methylation plays a very important role in the development of malignant tumors. The immune system is the key point in the progression of tumors, particularly in terms of tumor treatment and drug resistance. Tumor immunotherapy has now become a hot spot and a new approach for tumor treatment. However, as far as the stomach adenocarcinoma (STAD) is concerned, the in-depth research is still a gap in the m6A-associated immune markers. The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases is extremely important for our research, where we obtained gene mutation, gene expression data and relevant clinical information of STAD patients. Firstly, the samples from GEO were used as external validation groups, while the TCGA samples were divided into a training group and an internal validation group randomly. Using the way of Single factor COX-LASSO- and multi-factor Cox to construct the prognostic model. Then, all samples were subjected to cluster analysis to generate high and low expression groups of immune gene. Meanwhile, we also collected the correlation between these types and tumor microenvironment. On this basis, a web version of the dynamic nomogram APP was developed. In addition, we performed microenvironmental correlation, copy number variation and mutation analyses for model genes. The prognostic model for STAD developed here demonstrated a very strong predictive ability. The results of cluster analysis manifested that the immune gene low expression group had lower survival rate and higher degree of immune infiltration. Therefore, the immune gene low expression group was associated with lower survival rates and a higher degree of immune infiltration. Gene set enrichment analysis suggested that the potential mechanism might be related to the activation of immunosuppressive functions and multiple signaling pathways. Correspondingly, the web version of the dynamic nomogram APP produced by the DynNom package has successfully achieved rapid and accurate calculation of patient survival rates. Finally, the multi-omics analysis of model genes further enriched the research content. Interference of RAB19 was confirmed to facilitate migration of STAD cells in vitro, while its overexpression inhibited these features. The prognostic model for STAD constructed in this study is accurate and efficient based on multi-omics analysis and experimental validation. Additionally, the results of the correlation analysis between the tumor microenvironment and m6Ascore are the basics of further exploration of the pathophysiological mechanism in STAD.

Identification and subcellular localization analysis of membrane protein Ycf 1 in the microsporidian Nosema bombycis.

Microsporidia are obligate intracellular parasites that can infect a wide range of vertebrates and invertebrates including humans and insects, such as silkworm and bees. The microsporidium Nosema bombycis can cause pebrine in Bombyx mori, which is the most destructive disease in the sericulture industry. Although membrane proteins are involved in a wide range of cellular functions and part of many important metabolic pathways, there are rare reports about the membrane proteins of microsporidia up to now. We screened a putative membrane protein Ycf 1 from the midgut transcriptome of the N. bombycis-infected silkworm. Gene cloning and bioinformatics analysis showed that the Ycf 1 gene contains a complete open reading frame (ORF) of 969 bp in length encoding a 322 amino acid polypeptide that has one signal peptide and one transmembrane domain. Indirect immunofluorescence results showed that Ycf 1 protein is distributed on the plasma membrane. Expression pattern analysis showed that the Ycf 1 gene expressed in all developmental stages of N. bombycis. Knockdown of the Ycf 1 gene by RNAi effectively inhibited the proliferation of N. bombycis. These results indicated that Ycf 1 is a membrane protein and plays an important role in the life cycle of N. bombycis.