In recent years, the application of artificial intelligence (AI) in the field of biology has witnessed remarkable advancements. Among these, the most notable achievements have emerged in the domain of protein structure prediction and design, with AlphaFold and related innovations earning the 2024 Nobel Prize in Chemistry. These breakthroughs have transformed our ability to understand protein folding and molecular interactions, marking a pivotal milestone in computational biology. Looking ahead, it is foreseeable that the accurate prediction of various physicochemical properties of proteins—beyond static structure—will become the next critical frontier in this rapidly evolving field. One of the most important protein properties is thermodynamic stability, which refers to a protein’s ability to maintain its native conformation under physiological or stress conditions. Accurate prediction of protein stability, especially upon single-point mutations, plays a vital role in numerous scientific and industrial domains. These include understanding the molecular basis of disease, rational drug design, development of therapeutic proteins, design of more robust industrial enzymes, and engineering of biosensors. Consequently, the ability to reliably forecast the stability changes caused by mutations has broad and transformative implications across biomedical and biotechnological applications. Historically, protein stability was assessed via experimental methods such as differential scanning calorimetry (DSC) and circular dichroism (CD), which, while precise, are time-consuming and resource-intensive. This prompted the development of computational approaches, including empirical energy functions and physics-based simulations. However, these traditional models often fall short in capturing the complex, high-dimensional nature of protein conformational landscapes and mutational effects. Recent advances in machine learning (ML) have significantly improved predictive performance in this area. Early ML models used handcrafted features derived from sequence and structure, whereas modern deep learning models leverage massive datasets and learn representations directly from data. Deep neural networks (DNNs), graph neural networks (GNNs), and attention-based architectures such as transformers have shown particular promise. GNNs, in particular, excel at modeling spatial and topological relationships in molecular structures, making them well-suited for protein modeling tasks. Furthermore, attention mechanisms enable models to dynamically weigh the contribution of specific residues or regions, capturing long-range interactions and allosteric effects. Nevertheless, several key challenges remain. These include the imbalance and scarcity of high-quality experimental datasets, particularly for rare or functionally significant mutations, which can lead to biased or overfitted models. Additionally, the inherently dynamic nature of proteins—their conformational flexibility and context-dependent behavior—is difficult to encode in static structural representations. Current models often rely on a single structure or average conformation, which may overlook important aspects of stability modulation. Efforts are ongoing to incorporate multi-conformational ensembles, molecular dynamics simulations, and physics-informed learning frameworks into predictive models. This paper presents a comprehensive review of the evolution of protein thermodynamic stability prediction techniques, with emphasis on the recent progress enabled by machine learning. It highlights representative datasets, modeling strategies, evaluation benchmarks, and the integration of structural and biochemical features. The aim is to provide researchers with a structured and up-to-date reference, guiding the development of more robust, generalizable, and interpretable models for predicting protein stability changes upon mutation. As the field moves forward, the synergy between data-driven AI methods and domain-specific biological knowledge will be key to unlocking deeper understanding and broader applications of protein engineering.

ObjectiveTo investigate and analyze an outbreak of rotavirus infectious diarrhea in a prison in Chongqing Municipality, to provide a basis for adult rotavirus surveillance and prevention, and to explore the public health problems in special settings. MethodsA retrospective survey was conducted to collect and analyze data on individual cases with diarrheal disease on-site. The clinical characteristics, as well as the temporal, spatial and geographical distribution patterns of the epidemic were described. Multi-pathogen detection tests were conducted both on diarrhea cases and environmental samples, with viral genotyping performed on positive samples. A case-control analysis was performed to identify the causes of the outbreak, and an SEIR model was adopted to predict the outbreak trend and evaluate the effectiveness of interventions. ResultsA total of 65 cases were found among the inmates, with an attack rate of 2.03%. The predominant clinical manifestations included diarrhea (89.23%), watery stool (73.85%), and dehydration (18.46%). The epidemic curve indicated a “human-to-human” transmission pattern, with an average incubation period of 5‒6 days. The attack rates among chefs in the main canteen (80.00%, 8/10) and caterers (28.33%, 17/60) were significantly higher than those of other inmates （P<0.05）. Multi-pathogen polymerase chain reaction (PCR) testing detected positive for group A rotavirus, with the viral genotyping identified as G9P [8] strain. Factors such as unprotected "bare-handed" food distribution among cases with diarrhea (OR=9.512, 95%CI: 4.261‒21.234) and close contact with diarrhea cases (OR=3.656, 95%CI: 1.719‒7.778) were the possible cause of the outbreak. The SEIR model (r₀=5, α=0.3, β₁=0.08, β₂=0.04) was constructed using prison inmates as susceptible population, aiming at fitting the initial transmission trend of the outbreak, and the epidemic rate declined rapidly after intervention measures were implemented (r_t≈0). ConclusionThis rare rotavirus infection diarrhea outbreak among adults in confined settings suggests that the construction of public health prevention and control systems in prison may be overlooked. Cross infection during meal processing and distribution in the canteens of such settings is likely to be the cause of the outbreak. Given the potential neglect of public heath system construction in special settings, it is imperative to enhance the surveillance and monitoring of rotavirus and other intestinal multi-pathogens among adults, as well as the construction of public health prevention and control systems in these special settings.

Objective To evaluate the safety and efficacy of valve-in-valve transcatheter mitral valve replacement(ViV-TMVR)in patients with bioprosthetic valve degeneration who are at high surgical risk.Methods This study is a multi-center,retrospective cohort analysis of 20 consecutive patients who underwent transseptal ViV-TMVR using the Edwards SAPIEN 3 transcatheter heart valve(THV).The primary endpoints include technical success and procedural success,both defined according to the Mitral Valve Academic Research Consortium(MVARC)criteria,as well as mortality and functional change assessed based on New York Heart Association(NYHA)classification at 30-days and six months post-procedure.Clinical follow-up assessments are conducted at 30-days and six months.Results From February 2021 to October 2022,a total of 20 patients with symptoms of bioprosthetic valve degeneration were enrolled across nine sites in China.The patients had a mean age of(73.5±5.5)years,with 85.0％being females and 70.0％classified as NYHA class Ⅲ/Ⅳ.The study achieved a 100.0％technical success rate and a 90.0％procedural success rate finally.All patients remained alive during the 30-day follow-up period.However,six months post-intervention,two patients(10.0％)were re-hospitalized due to heart failure,and sadly,one of them(5.0％)died.None of the patients reported any adverse events related to ViV-TMVR during the follow-up period.Notably,there was a significant improvement in NYHA class compared to baseline(P=0.0004)at six-month follow-ups.Conclusions The transseptal ViV-TMVR technique proved to be highly successful and was associated with significant improvement in NYHA class function.These findings strongly suggest that it serves as a safe and efficient treatment alternative for high-risk patients suffering from bioprosthetic valve degeneration.

Objective:To provide novel genetic biomarkers for the diagnosis and treatment of sepsis,bioinformatics analysis was used to screen differentially expressed genes and identify Hub genes in sepsis.Methods:Gene Expression Omnibus(GEO)database was used to retrieve gene expression datasets of sepsis and screen for differentially expressed genes(DEGs).Protein-protein interaction(PPI)network analysis,Gene Ontology(GO)analysis,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were used to clarify the molecular mechanism of DEGs,and Hub genes were screened.Results:A total of 361 DEGs were identified,including 163 up-regulated genes and 198 down-regulated genes.Enrichment analysis revealed that these DEGs were primarily involved in antigen processing and presentation,T cell biology,cell adhesion molecules,and T cell receptor signaling pathways.CD4,TP53,PTPRC,LCK,ITGAM,ZAP70,CD247,CD2,CD3E,and HSP90AB1 were determined as optimal diagnostic biomarkers for sepsis.Conclusions:This study elucidated 10 Hub genes(CD4,TP53,PTPRC,LCK,ITGAM,ZAP70,CD247,CD2,CD3E,and HSP90AB1)as potential biomarkers for the diagnosis and treatment of sepsis.However,since the the generalizability of these Hub genes in patients with sepsis remains unvalidated,further experimental verification is still needed in the future.

Objective:To explore the application value of the integrated situational simulation teaching model based on the "Internet+"in obstetrics and gynecology clinical clerkship.Methods:Fifty-eight undergraduate students enrolled in 2019 who participated in the clinical clerkship at the Chengdu Fifth People's Hospital from June to September 2023 were selected and divided into a research group ( n=29) and a control group ( n=29) using the random number table method. The research group received the integrated situational simulation teaching model based on "Internet+", while the control group received traditional teaching methods. After clinical clerkship, theoretical tests, teacher evaluation, and questionnaire surveys were conducted to evaluate and compare the teaching effectiveness of the two groups of students. SPSS 23.0 was used for t test and chi-square test. Results:After receiving the integrated situational simulation teaching model based on "Internet+" for clinical clerkship, the research group achieved significantly higher theoretical test scores than the control group [(83.62±4.95) vs. (72.93±4.70), t=8.43, P<0.001]. The scores of all items in the mini-clinical evaluation exercise of the research group were higher than those of the control group ( P<0.01). According to the teaching feedback questionnaire, all students were satisfied with this new teaching model and reported that this model increased their interest in learning. Moreover, 86.21% (25/29) and 96.55% (28/29) of the students believed that the new model not only improved learning efficiency but also facilitated the integration of theory and practice. Conclusions:The integrated situational simulation teaching model based on "Internet+" is feasible in obstetrics and gynecology clinical clerkship and contributes to enhancing the quality of clinical clerkship teaching. This model is worthy of further research and promotion.

Objective To explore the current status of penile erection hardness and its influencing factors in patients with non-obstructive azoospermia.Methods From January to December 2024,450 patients with non-obstructive azoospermia were surveyed at the Reproductive Medicine Center of Hospital.A self-made general information questionnaire was used to collect their demographic data.The Erectile Hardness Scale(EHS)was employed to investigate the current status of their penile erection hardness,and a self-made questionnaire was utilized to explore the influencing factors.Results Among the 450 patients with non-obstructive azoospermia,during sexual intercourse,35.3%of the patients reported that their penile erection hardness could reach grade 4(normal state),54.5%reported that it only reached grade 3(sub-optimal state),9.3%reported that it only reached grade 2(slight penile erection),and 0.9%reported that it only reached grade 1(inability to achieve an erection).In the survey of satisfaction with sexual life quality,among the 450 patients,only 24.9%were very satisfied with their sexual life quality;57.3%were basically satisfied;9.6%considered it average;4.0%were dissatisfied;3.1%were very dissatisfied;and 1.1%had no sexual life.alcohol consumption(OR=2.393,95%CI:1.493～3.836),satisfaction with the quality of sexual life(OR=1.455,95%CI:1.118～1.894),educational attainment(OR=0.709,95%CI:0.549～0.917),and the sleep quality in the past month(OR=0.641,95%CI:0.452～0.907).Conclusions Clinical studies have shown that factors such as drinking habits,sexual life satisfaction,sleep quality,and educational attainment collectively influence the penile erection hardness in patients with non-obstructive azoospermia.Therefore,the medical team needs to customize personalized intervention plans and educational materials based on individual differences among patients.Through psychological counseling and lifestyle guidance,they can improve erectile function and the quality of sexual life,promote harmonious marital relationships,and enhance the overall life experience of the patients.

Objective:To develop a vasectomy intention scale (VIS) and evaluate its reliability and validity for assessing men's intentions toward vasectomy.Methods:Based on the Theory of Planned Behavior, the VIS was developed through a process that included literature review, panel discussions, expert consultations, and a pilot survey. A total of 264 men seeking vasectomy consultation at the Andrology Center, Department of Urology, the Third Xiangya Hospital of Central South University between December 2023 and December 2024 were recruited to assess reliability and validity of the VIS.Results:The VIS comprises 11 items across three dimensions: "background" factors, "stance and behavior" factors and "information" factors. The scale demonstrated satisfactory internal consistency (Cronbach's α=0.739). Correlations between each dimension and the total scale ("background" factors r=0.849, "stance and behavior" factors r=0.744, "information" factors r=0.440) exceeded inter-dimension correlations (ranging from 0.145 to 0.312), confirming robust construct validity. The vasectomy rates among men with different intention levels were 65.7% (92/140) in the high-intention group, 28.9% (33/114) in the moderate-intention group, and 0% (0/11) in the low-intention group, with a statistically significant difference (χ2=43.42, P<0.001). Conclusion:The VIS exhibits strong reliability and validity, serving as a validated instrument for measuring the strength of men's vasectomy intentions.

Objective To explore the current status of penile erection hardness and its influencing factors in patients with non-obstructive azoospermia.Methods From January to December 2024,450 patients with non-obstructive azoospermia were surveyed at the Reproductive Medicine Center of Hospital.A self-made general information questionnaire was used to collect their demographic data.The Erectile Hardness Scale(EHS)was employed to investigate the current status of their penile erection hardness,and a self-made questionnaire was utilized to explore the influencing factors.Results Among the 450 patients with non-obstructive azoospermia,during sexual intercourse,35.3%of the patients reported that their penile erection hardness could reach grade 4(normal state),54.5%reported that it only reached grade 3(sub-optimal state),9.3%reported that it only reached grade 2(slight penile erection),and 0.9%reported that it only reached grade 1(inability to achieve an erection).In the survey of satisfaction with sexual life quality,among the 450 patients,only 24.9%were very satisfied with their sexual life quality;57.3%were basically satisfied;9.6%considered it average;4.0%were dissatisfied;3.1%were very dissatisfied;and 1.1%had no sexual life.alcohol consumption(OR=2.393,95%CI:1.493～3.836),satisfaction with the quality of sexual life(OR=1.455,95%CI:1.118～1.894),educational attainment(OR=0.709,95%CI:0.549～0.917),and the sleep quality in the past month(OR=0.641,95%CI:0.452～0.907).Conclusions Clinical studies have shown that factors such as drinking habits,sexual life satisfaction,sleep quality,and educational attainment collectively influence the penile erection hardness in patients with non-obstructive azoospermia.Therefore,the medical team needs to customize personalized intervention plans and educational materials based on individual differences among patients.Through psychological counseling and lifestyle guidance,they can improve erectile function and the quality of sexual life,promote harmonious marital relationships,and enhance the overall life experience of the patients.

Objective:To investigate the differences in dosimetric parameters for target volumes and organs at risk (OARs), radiation doses to reconstructed tissues, and beam-on time in radiotherapy among helical tomotherapy (HT), volumetric modulated arc therapy (VMAT), and fixed-field intensity-modulated radiotherapy (F_IMRT) following immediate breast reconstruction for breast cancer, thereby providing a reference for the selection of clinical radiotherapy techniques.Methods:This study retrospectively investigated 15 breast cancer patients who underwent radiotherapy following modified radical mastectomy and immediate breast reconstruction at the Liuzhou Worker′s Hospital from August 2018 to July 2023. During target volume delineation, precautions were taken to avoid the reconstructed tissues, which were delineated separately. Customized HT, VMAT, and F_IMRT treatment plans were designed for each patient. The plans were categorized into the HT, VMAT, and F_IMRT groups based on different radiotherapy techniques employed. They were comparatively analyzed through one-way analysis of variance (ANOVA), with multiple comparisons further conducted in the case of significant differences.Results:Statistical analyses reveal significant differences in various parameters of target volumes among the three groups of plans ( F = 38.73, 14.95, 37.01, 48.05, 35.55, 22.56, 34.30, P < 0.05). Pairwise comparisons indicate that the maximum dose ( D2%), minimum dose ( D98%), mean dose ( Dmean), and the proportion of high-dose volumes within the target volume ( V107%and V110%) in both the HT and VMAT groups were significantly better than those in the F_IMRT group. The HT group demonstrated the optimal conformity index (CI), while the VMAT group displayed the superior homogeneity index (HI) compared to the other two groups. In terms of OAR, the V20 of the ipsilateral lung was the lowest in the HT group ( F = 14.31, P < 0.05) and the highest in the F_IMRT group ( F = 14.31, P < 0.05). However, the V5 and Dmean for both the ipsilateral and contralateral lungs in the HT group significantly surpassed those of the other groups ( F = 39.16, 31.91, P < 0.05). The mean dose Dmean ( F = 5.57, P < 0.05) of the contralateral breast was significantly reduced in the VMAT group compared to the other two groups. No statistically significant differences were observed for other OARs, including the heart, spinal cord PRV, thyroid, and humeral head ( P > 0.05). The radiation doses to reconstructed tissues ( Dmax, V53.5, Dmean) ascended in the order of HT, VMAT, and F_IMRT groups ( F = 17.69, 17.53, 15.11, P < 0.05). The HT and F_IMRT groups showed similar beam-on times ( P > 0.05), both exceeding that of the VMAT group by several folds ( F = 28.72, P < 0.05). Conclusions:The comparative analysis indicates that the three radiotherapy techniques exhibit distinct advantages and limitations, with F_IMRT demonstrating the least comprehensive advantage. HT can enhance the conformity of target volumes while reducing the overall radiation doses to reconstructed tissues and the crucial indicator V20 in the ipsilateral lung. VMAT demonstrates the highest treatment efficiency, yielding improved dose uniformity in the target volume and reduced radiation doses to the contralateral breast. It is advisable to prioritize HT or VMAT based on actual clinical conditions.

Objective:To analyze the clinical features and prognosis of acute B lymphoblastic leukemia (B-ALL) children carrying a TCF3: : PBX1 fusion gene and to evaluate the prognostic value of this gene.Methods:Retrospective cohort study.A total of 2 164 B-ALL children aged 0-18 years diagnosed and treated at 19 pediatric centers from October 2016 to June 2022 were enrolled.They were divided into the positive group and the negative group according to whether they carried a TCF3: : PBX1 fusion gene.The clinical characteristics, treatment response, adverse reactions, and prognosis of the 2 groups of patients were analyzed.The rank sum and Kruskal-Wallis tests were used to compare two and more than two groups of numerical variables, respectively.Fisher′s exact test was used to compare categorical variables.Results:Among the 2 164 patients, 116 (5.4%) were TCF3: : PBX1 positive, of which 70 patients were female, accounting for 60.3%.There were 840 female patients in the TCF3: : PBX1-negative group, accounting for 41.0%.There was a significant difference in the ratio of females between the TCF3: : PBX1-positive and TCF3: : PBX1-negative groups ( P<0.001).No significant difference was observed in age of onset between the two groups( P>0.05).The proportion of bone marrow naive cells [54.00 (14.00, 76.50)% vs.29.00 (3.00, 68.00)%], white blood cell counts [25.30 (10.46, 60.94)×10 9/L vs.9.03 (4.38, 30.73)×10 9/L] and hemoglobin counts [82.00(63.00, 101.00) g/L vs.74.00(60.00, 90.00) g/L] in the TCF3: : PBX1-positive group were significantly higher than those in the negative group at the onset (all P<0.05).In terms of treatment response, the proportion of peripheral blood naive cells on Day 8 in the TCF3: : PBX1-positive group was significantly higher than that in the negative group [2.00 (0, 9.00)% vs.0 (0, 2.00)%, P<0.001].The proportion of minimal residual disease <0.1% on Day 15 in the TCF3: : PBX1-positive group was significantly higher than that in the negative group ( P=0.038).There were no significant differences in cumulative recurrence rate, treatment-related mortality (TRM), and overall survival (OS) between the TCF3: : PBX1-positive group and TCF3: : PBX1-negative group (all P>0.05).The cumulative recurrence risk of TCF3: : PBX1-positive patients was 9.646 times higher than that of ETV6: : RUNX1-positive patients with better prognosis( HR=9.646, 95% CI: 1.026-90.700, P=0.047).There were no significant differences in TRM and OS between TCF3: : PBX1-positive and ETV6: : RUNX1-positive patients (all P>0.05).A significant enrichment of PAX5 mutations was detected in TCF3: : PBX1-positive patients.Among the 7 high-risk TCF3: : PBX1-positive patients in a single center, 4 patients had PAX5 mutations, and this proportion was significantly higher than that in other patients ( P<0.001). Conclusions:B-ALL children carrying a TCF3: : PBX1 fusion gene have a high remission rate and good long-term prognosis after intensive chemotherapy.It is suggesting that TCF3: : PBX1-positive B-ALL patients should be rated at intermediate risk to receive intensive chemotherapy.