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.

Objective To analyze and evaluate the implementation effect of tuberculosis prevention and control program in Wuhan, and to provide reference for scientific formulation of tuberculosis prevention and control measures. Methods Using the National Tuberculosis Information Management System, descriptive statistical analysis was carried out on the medical record information of pulmonary tuberculosis patients registered in Wuhan , 2016 - 2021. Results A total of 34 937 cases of pulmonary tuberculosis were registered in Wuhan , with an average annual incidence rate of 49.85/100 000. The incidence rate showed a downward trend year by year, with a statistically significant difference in 2016—2021 (χ²_trend = 708.387, P<0.001). The patients mainly came from referrals, accounting for 71.86%, and the proportion of referrals varied significantly among different years (χ²=355.541, P<0.001). The diagnosis type was mainly pathogenic negative, accounting for 49.12%. The proportion of pathogenic negative had statistically significant difference among different years (χ²=1 354.830, P<0.001). The proportion of patients cured and completed the course of treatment reached 93.98%, with statistically significant differences in the proportions among different years (cured, χ²=1 080.252, P<0.001; completed the treatment course, χ²= 933.655, P<0.001). The sputum examination rate of newly diagnosed patients in each year reached over 90%, and the overall completion rate reached over 95%. The proportion of positive pathogens showed an increasing trend year by year. Conclusion The overall epidemic situation of tuberculosis in Wuhan is declining year by year, and tuberculosis prevention and control work has achieved remarkable results. Active screening in key areas and populations should be strengthened, and prevention and control strategies should be formulated by emphasizing the key and difficult points.

Objective:To compare the short-term effect of the arthroscopic reconstruction of anterior cruciate ligament (ACL) by femoral tunnel positioning through the tendon incision (TI) technique and anteromedial (AM) technique.Methods:Between December 2015 and March 2017, 176 patients with ACL injuries underwent arthroscopic ACL reconstruction at the First Affiliated Hospital of USTC were analyzed. According to the method of localization of femoral tunnel, they were divided into two groups: TI group (localization of femoral tunnel by TI for reconstruction of ACL) and AM group (localization of femoral tunnel by AM for reconstruction of ACL). There were 87 patients in the TI group (63 males and 24 females) with an age of 32.8±9.4 years (range, 19-51 years) and a body mass index (BMI) of 24.8±6.3 kg/m 2 (range, 18.1-31.7 kg/m 2), including 9 obese patients (BMI>28 kg/m 2). There were 89 patients in the AM group (59 males and 30 females) with an age of 36.7±13.0 years (range, 17-56 years) and a BMI of 25.7±5.8 kg/m 2 (range, 18.9-31.6 kg/m 2), including 11 obese patients. To evaluate whether the drill was in contact with the articular surface of the medial condyle of the femur, whether the anterior angle of the medial meniscus was damaged, the sagittal and coronal angles of the femoral tunnel and the total length of the femoral tunnel. The anterior drawer test, Lachmann test, pivot shift test, knee flexion angle (KFA), Lysholm score, International Knee Documentation Committee (IKDC) score and the incidence of postoperative complications were compared between the two groups. At the same time, the time of taking out tendon, operation time, hospital stay and recovery time were compared between two groups of obese people. Results:In the TI group, the guide pin could reach all areas of the medial wall of the lateral femoral condyle from the inferior cartilage edge to the top of the intercondylar fossa, the drill bit had no contact with the medial femoral condyle (MFC), and the anterior angle of the medial meniscus was not damaged. In AM Group, 3 cases of MFC and 1 case of anterior horn of medial meniscus were injured in order to get the best position of femoral tunnel. The angle of femoral tunnel in sagittal plane (50.2°±3.2°) and coronal plane (46.1°±5.8°) in experimental group was notably larger than that in control group (45.6°±5.4°, 38.3°±4.7°), the difference was statistically significant ( P<0.05). However, there was no significant difference in the length of femoral tunnel between the two groups (38.2±3.2 mm and 37.7±2.5 mm, P>0.05). All patients were followed up for an average of 8.3±2.1 (range, 3-12) months. The positive rates of anterior drawer test, Lachmann test and pivot shift test were not significantly different between the two groups at 3, 6, 9 and 12 months after operation ( P>0.05). The KFA, Lysholm and IKDC scores in both groups were significantly increased after operation ( P<0.05), but there was no significant difference of above indexes between the two groups ( P>0.05). The overall complication rate in AM group (14.6%, 13/89) was significantly higher than that in TI group (6.9%, 6/87), the difference was statistically significant ( P<0.05). Additionally, the tendon harvesting (15.0±0.2 min vs. 26.0±0.2 min, P<0.05) and operation time (2.0±0.3 h vs. 3.0±0.4 h, P<0.05) were significantly shortened among obese patients in experimental group compared with those in control group. Conclusion:The TI technique demonstrates comparable effectiveness to the AM technique in ACL reconstruction through femoral tunnel positioning, with the added advantage of safer femoral tunnel localization. This approach minimizes the risk of iatrogenic injuries to the articular cartilage of the medial condyle of the femur and the anterior horn of the medial meniscus. Furthermore, the TI technique reduces tendon removal time, particularly beneficial for obese patients.

Objective To investigate the impact of human leukocyte antigen(HLA)functional epitope mismatch(EM)on the efficacy of platelet transfusion in patients with hematological diseases.Methods HLA genotyping was performed on platelet donors and patients with hematological diseases who applied for platelet serological cross-matching and HLA antigen matching from June 2021 to June 2023 by PCR-SBT method.HLA platelet matching was based on the principle of CREG to se-lect donors for patients.HLA Matchmaker 4.0 software was used to analyze donor-recipient HLA EM information.The expres-sion level and gene distribution of related HLA functional epitope(Eplet)were searched from the international HLA Epitope registry website(www.Epregistry.com.br).Retrospective analysis was conducted on clinical platelet transfusion data to evalu-ate the impact of HLA EM on platelet transfusion effectiveness.Results Platelet transfusion efficacy showed no correlation with gender and age,but it was associated with platelet matching strategy.When the total number of HLA EMs was less than 20,a lower total number of donor-recipient HLA EMs resulted in higher platelet transfusion efficiency(χ2=19.311,P=0.001)and higher average value of 24 h corrected count increment(CCI)(F=7.737,P<0.001).The total number of donor-recipient HLA EMs had negative correlation with actual 24 h CCI(Rho=-0.322,P<0.001).Further statistical analysis re-vealed that 17 Eplets were related to the effectiveness of platelet transfusion.The locus distribution of 17 Eplets might be u-nique to HLA-A(17.6%)or-B(64.7%)or shared between HLA-A and-B(17.6%),and its expression may be high(58.8%)or intermediate(41.2%).Conclusion The total number of donor-recipient HLA EMs is a crucial factor influencing platelet transfusion effectiveness,and several HLA Eplets associated with this effectiveness have been identified.

Objective To explore the value of droplet digital polymerase chain reaction(ddPCR)in the etiological diagnosis of severe acute pancreatitis(SAP)patients with suspected bloodstream infection(BSI).Methods SAP patients admitted to the department of critical care medicine in a hospital July to September 2022 were enrolled.When BSI was suspected,venous blood was collected for both ddPCR detection and blood culture(BC)with antimi-crobial susceptibility testing(AST)simultaneously.The time required for two detection methods was recorded,and the detection results of ddPCR and BC were compared.The etiological diagnostic efficacy of ddPCR was calculated,and the correlation between the value of pathogen load detected by ddPCR and the level of infection parameters was explored.Results A total of 22 patients were included in the analysis,and 52 venous blood specimens were collec-ted for detection.BC revealed 17 positive specimens(32.7％)and 29 pathogenic strains,while ddPCR showed 41 positive specimens(78.8％)and 73 pathogenic strains.Detection time required for ddPCR was significantly lower than that of BC([0.16±0.03]days vs[5.92±1.20]days,P＜0.001).Within the detection range of ddPCR and taking BC results as the gold standard,the sensitivity and specificity of ddPCR were 80.0％and 28.6％,respective-ly.With the combined assessment of BSI based on non-blood specimen microbial evidence within a week,the sensi-tivity and specificity of ddPCR detection increased to 91.9％and 76.9％,respectively.ddPCR detected resistance genes of blaKPC,blaNDM/IMP,VanA/VanM,and mecA from 19,9,6,and 5 specimens,respectively.Correlation analysis showed a positive correlation between pathogen load and levels of C-reactive protein as well as procalcitonin(r=0.347,0.414,P＜0.05).Conclusion As a supplementary detection method for BC in BSI diagnosis,ddPCR has the advantages of higher sensitivity and shorter detection time,and is worthy of further exploration in clinical application.

A novel series of 2-aryl substituted benzothiopyranone compounds was designed and synthesized based on our previously obtained benzothiopyranone scaffold with significant antituberculosis activity. All target compounds were evaluated for their antimycobacterial activity and preliminary druggability was subsequently investigated for some selected compounds with good activity. The results indicated that most compounds showed good activity against Mycobacterium tuberculosis H₃₇Rv. Among them, compounds 8g, 8h, 8q and 9f showed potent activity with MIC ranged from 0.2 to 0.4 μg·mL^-1. Furthermore, some active compounds exhibited low cytotoxicity and cardiotoxicity risk. It is worth noting that compounds 8h and 8q with good liver microsome stability and low inhibition of CYPs 3A4/5 and 2C9 were suitable for combination drug regimen to treat tuberculosis.

Purpose/Significance The establishment of a specialized database for children with attention deficit hyperactivity disorder(ADHD)aims to address the issues of low data usage efficiency caused by scattered and low-quality diagnostic data.Method/Process Based on clinical diagnostic data of children with ADHD from March 2020 to April 2024,natural language processing(NLP)techniques are utilized to realize the post-structuring of the data.Furthermore,standardization of the database is performed according to industry standards and treatment guidelines.Result/Conclusion The specialized database for children with ADHD is established,comprising data from 75 835 ADHD children and 280 000 clinical visits.This database supports various applications such as the analysis of basic character-istics of the ADHD population,prediction model construction,resource allocation optimization,and clinical pathway optimization.

Objective:To evaluate the safety and efficiency of China-made Toumai Robot-assisted laparoscopic radical prosta-tectomy(LRP).Methods:This study included 40 cases of PCa treated from January 2023 to May 2023 by robot-assisted LRP with preservation of the bladder neck and maximal functional urethral length,15 cases with the assistance of Toumai Robot(the TMR group)and the other 25 with the assistance of da Vinci Robot as controls(the DVR group).We recorded the docking time,laparo-scopic surgery time,vesico-urethral anastomosis time,intraoperative blood loss and postoperative urinary continence,and compared them between the two groups.Results:Operations were successfully completed in all the cases.No statistically significant differ-ences were observed between the TMR and DVR groups in the docking time(6 min vs 5 min,P＞0.05)or intraoperative blood loss(200 ml vs 150 ml,P＞0.05).The TMR group,compared with the DVR group,showed a significantly longer median laparoscopic surgery time(146 min vs 130 min,P＜0.05)and median vesico-urethral anastomosis time(19 min vs 16 min,P＜0.05).There were no statistically significant differences between the TMR and DVR groups in the rates of urinary continence recovery immediately af-ter surgery(60.0％[9/15]vs 64.0％[16/25],P＞0.05)or at 1 month(80.0％[12/15])vs(76.0％[19/25],P＞0.05),3 months(93.3％[14/15])vs(92.0％[23/25],P＞0.05)and 6 months postoperatively(100％[15/15])vs(96％[24/25],P＞0.05).Conclusion:China-made Toumai? Robot surgical system is safe and reliable for laparoscopic radical prosta-tectomy,with satisfactory postoperative recovery of urinary continence.

Objective:To analyze the F10 gene mutations in a Chinese pedigree affected with the deficiency of the hereditary coagulation factor X (FX), resulting from a new deletion mutation, and to study the associated molecular pathogenesis.Methods:Next generation sequencing (NGS) was performed to screen the genetic mutations in the proband which were then verified by Sanger sequencing. The FX activity (FX∶C) of probands and their family members was detected using the blood clotting method, and the mutation sites of the family members were analyzed using Sanger sequencing. The pathogenicity of the mutation site was predicted by using the online bioinformatics software, Mutation Taster. The SWISS-MODEL software was used for stimulating the three-dimensional models of the wild-type and mutant proteins for analyzing the influence of the mutation site on the structure and function of the proteins, and for analyzing the difference between the catalytic residues of the wild-type and the mutant proteins. The level of the F10 gene mRNA was quantitatively analyzed by qRT-PCR (quantitative reverse transcription polymerase chain reaction) method by constructing plasmids, transfecting human embryonic kidney 293T cells (HEK 293T), and analyzing the splicing of the mutated site by RT-PCR method. The levels of FⅩ∶Ag in cell lysates and cell culture media (both inside and outside the cells) were detected by the ELISA (enzyme linked immunosorbent assay) method.Results:A medium-grade factor X deficiency with a 36.42% FⅩ∶C ratio was detected in the proband by the coagulation method. NGS analysis demonstrated a heterozygous deletion mutation in exon 8:c.902_919del (p.Ala301_Glu306del) in the proband. Sanger sequencing analysis indicated that some members of the family (mother and grandfather) were also carriers of the corresponding deletion mutation. Online bioinformatics software predicted the pathogenic nature of the c.902_919del mutation, with a pathogenic score of 0.999. The 3D protein structure model analysis indicated that the c.902_919del mutation resulted in the disappearance of a segment of β-fold in the protein structure, thereby shortening the preceding segment of the β-fold and a subsequent loss of hydrogen bonds between adjacent amino acids with no significant difference in the side chain conformation of the key catalytic residues compared to the wild-type. mRNA splicing analysis indicated the absence of alternative splicing changes in the mutation, and qRT-PCR results indicated the absence of a statistically significant difference between the mRNA levels of F10 gene and wild-type mRNA in cells expressing c.902_919del mutant. The ELISA results indicated that there was no statistically significant difference in the FX∶Ag levels of the mutant cell culture medium and the lysate.Conclusions:In this pedigree, the heterozygous mutation in exon 8 of F10 gene (c.902_919del, p.Ala301_Glu306del) caused the hereditary factor Ⅹ deficiency.

Objective To investigate the influence of sodium bicarbonate Ringer's solution and compound Ringer's solution on patients'acid-base balance and liver and kidney function during controlled low central venous pressure(CLCVP)hepatectomy.Methods A total of 60 patients with primary liver cancer who underwent CLCVP hepatectomy in the Affiliated Cancer Hospital of Zhengzhou University from June 2020 to June 2022 were selected and randomly divided into the sodium bicarbonate Ringer's solution group and compound Ringer's solution group,with 30 patients in each group.Patients in the sodium bicarbonate Ringer's solution group were infused with sodium bicarbonate Ringer's solution,while patients in the compound Ringer's solution group were infused with compound Ringer's solution after anesthesia induction and before liver parenchymal disconnection.The pH value,base excess(BE)and blood lactic acid(Lac)levels were detected using the blood gas analyzer in the two groups before anesthesia induction(T,),at the beginning of surgery(T2),2 h during surgery(T3),4 h during surgery(T4),and at the end of surgery(T5),respectively.The levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood urea nitrogen(BUN),and serum creatinine(Scr)were detected using the automatic biochemical analyzer 24 h before and 24 h after surgery,respectively.The body mass,surgical time,intraoperative infusion volume,urine volume,changes in body acid-base balance during surgery,and changes in liver and kidney function before and after surgery between the two groups were compared.Results There was no statistically significant difference in body mass,surgical time,intraoperative infusion volume,and urine volume between the two groups of patients(P＞0.05).At T,,there was no statistically significant difference in pH value,BE and Lac levels between the two groups of patients(P＞0.05).Compared to T,,the pH value in the compound Ringer's solution group increased at T2 and decreased at T3,T4 and T5,while BE and Lac levels from T2 to T5 showed an increasing trend(P＜0.05);compared to T,,the pH value in the sodium bicarbonate Ringer's solution group increased at T2 and decreased at T3,BE and Lac levels increased at T2 and T3(P＜0.05),and there was no statistically significant difference in pH value,BE and Lac levels at T4 and T5 compared to T1(P＞0.05).At T5,the pH value in the sodium bicarbonate Ringer's solution group was significantly higher than that in the compound Ringer's solution group,while the BE and Lac levels were significantly lower than those in the compound Ringer's solution group(P＜0.05).There was no statisti-cally significant difference in the levels of ALT,AST,BUN and Scr 24 hours before surgery between the two groups of patients(P＞0.05);compared to 24 hours before surgery,there was no significant change in the levels of ALT,AST,BUN and Scr 24 hours after surgery(P＞0.05);there was no statistically significant difference in the levels of ALT,AST,BUN and Scr 24 hours after surgery between the two groups(P＞0.05).Conclusion Compared to compound Ringer's solution,sodium bicarbonate Ringer's solution has a smaller impact on the internal environment of the body during the CLCVP hepatectomy and can effectively maintain the stability of the body's acid-base balance.Both the two solutions have no significant impact on the patient's liver and kidney function.