In recent years, with the continuous advancement of related research, the clinical value of clot waveform analysis (CWA) in the diagnosis and management of thrombotic diseases has become increasingly prominent. As a dynamic coagulation monitoring technology based on optical principles, CWA overcomes the limitations of traditional coagulation tests (e.g., APTT, PT), which rely on single time-point parameters, in identifying hypercoagulable states, predicting thrombotic risk, and monitoring anticoagulant efficacy. By analyzing the kinetic profiles of the coagulation cascade, CWA provides multidimensional insights. This article elucidates the theoretical basis and principles of CWA, systematically reviews its applications in arterial/venous thrombosis, other hypercoagulability-related disorders, and anticoagulation therapy monitoring, and synthesizes recent advances of CWA for thrombotic diseases. Specifically, multiple studies demonstrate that CWA-APTT parameters (e.g., peak height Min1, Min2, Delta) can sensitively detect hypercoagulability. Combining CWA with the Padua score significantly enhances the predictive power for venous thromboembolism (VTE) risk assessment. CWA shows clinical utility in evaluating hypercoagulability in patients with acute myocardial infarction (AMI) and acute cerebral infarction (ACI), where parameters such as Min1, Min2, and Max2 exhibit greater sensitivity than conventional APTT. These metrics may predict AMI complications and guide clinical management. Additionally, CWA demonstrates value in diverse scenarios including pregnancy, inflammation-associated hypercoagulability (e.g., COVID-19, Kawasaki disease), and rare thrombotic conditions (e.g., chronic spontaneous urticaria). Beyond diagnostic and risk-stratification advantages, CWA serves as a novel tool for personalized anticoagulation monitoring. Its derivative technology, clot-fibrinolysis waveform analysis (CFWA), extends applications to fibrinolysis assessment—aiding in identifying coagulation status in deep vein thrombosis (DVT) patients, tracking coagulation/fibrinolysis dynamics in COVID-19, and evaluating efficacy of anticoagulant/antifibrinolytic therapies. Nevertheless, despite its unique strengths, challenges such as device dependency, insufficient standardization, and heterogeneity in parameter interpretation hinder widespread clinical adoption, necessitating further investigation. Future directions include establishing multidimensional thrombotic risk assessment systems integrating CWA and developing AI-powered automated CWA analysis platforms to enhance clinical accessibility.

Objective:To establish a method for quantitative detection of BK virus (BKV) nucleic acid using microfluidic droplet one-pot Recombinase polymerase amplification-clustered regularly interspaced short palindromic repeats, RPA-CRISPR/Cas13a.Methods:A methodology establishment study. Plasma or urine samples were collected from 50 post renal transplant patients and 40 healthy individuals in Renji Hospital, Shanghai Jiao Tong University School of Medicine, from May 16 to August 31, 2024, and the viral genomic DNA was extracted. In the renal transplant group, there were 28 males and 22 females aged (46.1±11.6) years; in the healthy control group, there were 21 males and 19 females aged (45.6±11.3) years. Three crRNAs were designed for screening the optimal crRNA in CRISPR/Cas13a reaction taking PCR product as templet. Two pairs of RPA primers targeting BKV sequences were designed and selected an optimal primer set from four combinations for RPA-CRISPR/Cas13a reaction. Microfluidic droplet generation chip was designed and fabricated for rapid quantitative detection of BKV nucleic acids, which was combined with a droplet reader. BKV plasmid standards were utilized to assess the detection sensitivity of the one-pot assay. The CRISPR droplet one-pot assay and qPCR were utilized to detect the BKV load in the above samples, respectively. The methodological consistency was then analyzed, and Kappa coefficients of the results were calculated using the Kappa test, and the correlation between the two methods was evaluated using linear regression analysis. Results:The RPA-CRISPR/Cas13a reaction system was combined with a droplet chip to establish a CRISPR microfluidic droplet one-pot assay for BKV detection. The sensitivity of the CRISPR microfluidic droplet one-pot assay for detecting BKV was 10 copies/ml, which surpassed that of qPCR (100 copies/ml) without cross-reaction with JC virus. Methodological consistency analysis demonstrated that the Kappa coefficient of the two methods was 0.96, and the coefficient of determination R 2 was 0.95. Conclusion:In this study, a novel BKV detection technology based on the one-pot CRISPR droplet assay was successfully established.

Objective:To establish a one-pot lyophilized detection system based on recombinase polymerase amplification (RPA) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas13a) technology for the rapid diagnosis of cytomegalovirus (CMV) infection in liver transplantation recipients.Methods:This study is a methodology study. CRISPR RNA (crRNA) and RPA primers were designed targeting the CMV gene sequence. Optimal RPA primer sets were screened to establish the RPA-CRISPR/Cas13a-based CMV detection system. The limit of detection (LOD) was evaluated using gradient-diluted CMV plasmid standards. Cross-reactivity was assessed using genomic DNA from common opportunistic viruses in organ transplant recipients. Lyophilized reagents were validated with CMV-negative and positive samples. P-values were computed using two-sample t-tests for pairwise comparisons and one-way ANOVA for multi-group analyses to assess fluorescence value differences. Subsequently, lyophilized reagents were employed to detect 22 plasma samples from liver transplantation recipients collected at Renji Hospital, Shanghai Jiao Tong University School of Medicine, from June 3, 2024, to May 31, 2025. The test results were then compared with those obtained using quantitative real-time polymerase chain reaction (qPCR). Consistency between the two methods was evaluated using the Kappa coefficient calculated by Kappa test.Result:The established RPA-CRISPR/Cas13a system achieved a detection sensitivity of 1 copy/reaction and exhibited no cross-reactivity with other common opportunistic viruses in organ transplantation. Lyophilized RPA-CRISPR/Cas13a reagents demonstrated performance equivalent to non-lyophilized reagents. Concordance between lyophilized reagent detection and qPCR results for 22 clinical samples was 100% (22/22).Conclusion:A lyophilized CMV detection method based on RPA-CRISPR/Cas13a technology was successfully developed and validated for convenient diagnosing CMV infection in liver transplant recipients.

Objective To investigate the influencing factors of nutritional risk in patients with radia-tion-induced oral mucositis and establish a predictive model.Methods A total of 126 patients with nasopha-ryngeal carcinoma and radiation-induced oral mucositis who visited the General Hospital of Shaoxing Second Hospital Affiliated Healthcare Group,Dongpu Street Community Health Service Center in Yuecheng District,Shaoxing City,and the First Affiliated Hospital of Zhejiang University School of Medicine from January 2020 to January 2023 were selected as the model group and divided into the nutritional risk group and the nutrition-ally normal group according to the presence or absence of nutritional risk.Meanwhile,54 patients with radia-tion-induced oral mucositis due to nasopharyngeal carcinoma from February 2023 to March 2024 were selected as the validation group.Logistic regression was used to establish a risk prediction model,and a nomogram was developed and validated.Results Multivariate analysis showed that digestive discomfort symptoms,dietary structure score,and anxiety were factors influencing nutritional risk in patients with radiation-induced oral mucositis.In the model group,the area under the ROC curve was 0.992(95%CI:0.977 to 1.000),the maxi-mum Youden index was 0.961,corresponding specificity was 0.961,sensitivity was 1.000,χ2=4.668,P=0.792.In the validation group,the area under the ROC curve was 0.948(95%CI:0.897 to 0.999),the maxi-mum Youden index was 0.742,corresponding specificity was 0.742,sensitivity was 1.000,χ2=4.438,P=0.816.The model demonstrated good predictive ability.Conclusion A predictive model for nutritional risk factors in patients with radiation-induced oral mucositis is successfully constructed and can provide clinical reference.

Objective:To observe the clinical efficacy of DING's Tuina(Chinese therapeutic massage)plus Western medication in treating rotator cuff injury(RCI)and its impact on pain,shoulder function,and shoulder muscle strength in the patients.Methods:Sixty-two RCI patients were divided into an observation group and a control group using a simple randomization method,each consisting of 31 cases.Both groups of patients were prescribed oral celecoxib capsules.In addition,DING's Tuina treatment was offered to the observation group.After 6 weeks of treatment,the two groups were observed for changes in the Constant-Murley score(CMS)for shoulder joint function,the short-form McGill pain questionnaire(SF-MPQ)score,and peak torque(PT)and peak torque to body mass ratio(PT/BM)at angular velocities 120 °/s and 90 °/s as well as the peak torque ratio(PTR)of agonist muscles to antagonist muscles[including adductors to abductors(ADD/ABD)and flexors to extensors(F/E)].Results:After treatment,both groups presented decreased SF-MPQ scores and increased CMS,and the intra-group differences were statistically significant(P<0.05);the observation group had a lower SF-MPQ score and a higher CMS than the control group,showing statistical significance(P<0.05).After the intervention,the ADD/ABD decreased in the control group at the angular velocity 120 °/s in abduction,and the intra-group change was statistically significant(P<0.05);in the observation group,the PT and PT/BM increased compared to the baseline at angular velocities 120 °/s and 90 °/s in abduction,adduction,forward flexion,and backward extension,showing statistical significance(P<0.05).After treatment,the PT and PT/BM at angular velocities 120 °/s and 90 °/s and ADD/ABD at 120 °/s were higher in the observation group than in the control group,and the between-group differences were statistically significant(P<0.05).Conclusion:DING's Tuina combined with oral celecoxib capsules can reduce pain,improve the joint's motor function,and enhance the strength of shoulder muscle groups in RCI patients,and this treatment has a positive impact on the stability when performing shoulder joint abduction and adduction.

Objective:To observe the clinical efficacy of DING's Tuina(Chinese therapeutic massage)plus Western medication in treating rotator cuff injury(RCI)and its impact on pain,shoulder function,and shoulder muscle strength in the patients.Methods:Sixty-two RCI patients were divided into an observation group and a control group using a simple randomization method,each consisting of 31 cases.Both groups of patients were prescribed oral celecoxib capsules.In addition,DING's Tuina treatment was offered to the observation group.After 6 weeks of treatment,the two groups were observed for changes in the Constant-Murley score(CMS)for shoulder joint function,the short-form McGill pain questionnaire(SF-MPQ)score,and peak torque(PT)and peak torque to body mass ratio(PT/BM)at angular velocities 120 °/s and 90 °/s as well as the peak torque ratio(PTR)of agonist muscles to antagonist muscles[including adductors to abductors(ADD/ABD)and flexors to extensors(F/E)].Results:After treatment,both groups presented decreased SF-MPQ scores and increased CMS,and the intra-group differences were statistically significant(P<0.05);the observation group had a lower SF-MPQ score and a higher CMS than the control group,showing statistical significance(P<0.05).After the intervention,the ADD/ABD decreased in the control group at the angular velocity 120 °/s in abduction,and the intra-group change was statistically significant(P<0.05);in the observation group,the PT and PT/BM increased compared to the baseline at angular velocities 120 °/s and 90 °/s in abduction,adduction,forward flexion,and backward extension,showing statistical significance(P<0.05).After treatment,the PT and PT/BM at angular velocities 120 °/s and 90 °/s and ADD/ABD at 120 °/s were higher in the observation group than in the control group,and the between-group differences were statistically significant(P<0.05).Conclusion:DING's Tuina combined with oral celecoxib capsules can reduce pain,improve the joint's motor function,and enhance the strength of shoulder muscle groups in RCI patients,and this treatment has a positive impact on the stability when performing shoulder joint abduction and adduction.

Objective:To establish a method for quantitative detection of BK virus (BKV) nucleic acid using microfluidic droplet one-pot Recombinase polymerase amplification-clustered regularly interspaced short palindromic repeats, RPA-CRISPR/Cas13a.Methods:A methodology establishment study. Plasma or urine samples were collected from 50 post renal transplant patients and 40 healthy individuals in Renji Hospital, Shanghai Jiao Tong University School of Medicine, from May 16 to August 31, 2024, and the viral genomic DNA was extracted. In the renal transplant group, there were 28 males and 22 females aged (46.1±11.6) years; in the healthy control group, there were 21 males and 19 females aged (45.6±11.3) years. Three crRNAs were designed for screening the optimal crRNA in CRISPR/Cas13a reaction taking PCR product as templet. Two pairs of RPA primers targeting BKV sequences were designed and selected an optimal primer set from four combinations for RPA-CRISPR/Cas13a reaction. Microfluidic droplet generation chip was designed and fabricated for rapid quantitative detection of BKV nucleic acids, which was combined with a droplet reader. BKV plasmid standards were utilized to assess the detection sensitivity of the one-pot assay. The CRISPR droplet one-pot assay and qPCR were utilized to detect the BKV load in the above samples, respectively. The methodological consistency was then analyzed, and Kappa coefficients of the results were calculated using the Kappa test, and the correlation between the two methods was evaluated using linear regression analysis. Results:The RPA-CRISPR/Cas13a reaction system was combined with a droplet chip to establish a CRISPR microfluidic droplet one-pot assay for BKV detection. The sensitivity of the CRISPR microfluidic droplet one-pot assay for detecting BKV was 10 copies/ml, which surpassed that of qPCR (100 copies/ml) without cross-reaction with JC virus. Methodological consistency analysis demonstrated that the Kappa coefficient of the two methods was 0.96, and the coefficient of determination R 2 was 0.95. Conclusion:In this study, a novel BKV detection technology based on the one-pot CRISPR droplet assay was successfully established.

Objective:To establish a one-pot lyophilized detection system based on recombinase polymerase amplification (RPA) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas13a) technology for the rapid diagnosis of cytomegalovirus (CMV) infection in liver transplantation recipients.Methods:This study is a methodology study. CRISPR RNA (crRNA) and RPA primers were designed targeting the CMV gene sequence. Optimal RPA primer sets were screened to establish the RPA-CRISPR/Cas13a-based CMV detection system. The limit of detection (LOD) was evaluated using gradient-diluted CMV plasmid standards. Cross-reactivity was assessed using genomic DNA from common opportunistic viruses in organ transplant recipients. Lyophilized reagents were validated with CMV-negative and positive samples. P-values were computed using two-sample t-tests for pairwise comparisons and one-way ANOVA for multi-group analyses to assess fluorescence value differences. Subsequently, lyophilized reagents were employed to detect 22 plasma samples from liver transplantation recipients collected at Renji Hospital, Shanghai Jiao Tong University School of Medicine, from June 3, 2024, to May 31, 2025. The test results were then compared with those obtained using quantitative real-time polymerase chain reaction (qPCR). Consistency between the two methods was evaluated using the Kappa coefficient calculated by Kappa test.Result:The established RPA-CRISPR/Cas13a system achieved a detection sensitivity of 1 copy/reaction and exhibited no cross-reactivity with other common opportunistic viruses in organ transplantation. Lyophilized RPA-CRISPR/Cas13a reagents demonstrated performance equivalent to non-lyophilized reagents. Concordance between lyophilized reagent detection and qPCR results for 22 clinical samples was 100% (22/22).Conclusion:A lyophilized CMV detection method based on RPA-CRISPR/Cas13a technology was successfully developed and validated for convenient diagnosing CMV infection in liver transplant recipients.

Objective:To optimize the processing technology of vinegar-Corni Fructus using Box Behnken design and entropy weight method.Methods:The optimal processing parameters of vinegar-Corni Fructus were optimized by Box-Behnken design. 5-Hydroxymethylfurfural, Nuxaside, Monosanthin, and appearance character were used as comprehensive evaluation indexes. The entropy weight method was used to calculate the weights of each index, taking Design Expert 12.0 for data analysis.Results:The optimal process for vinegar-Corni Fructus as follow: The ratio of excipients was 15%, the simmering time was 2 h, and the steaming time was 8 h.Conclusion:The optimal vinegar-Corni Fructus processing is stable and feasible, which can be used for the production of processed prescriptions.

Immune checkpoint inhibitor(ICI)associated diabetes mellitus(DM)a rare specific type of diabetes,usually developed after the treatment of programmed cell death protein 1(PD-1)inhibitor or programmed cell death ligand 1(PD-L1)inhibitor.We reported 4 cases of DM after ICI use.These patients had a subacute onset,6 to 17 months after the first use of the drug,with diabetic autoantibodies negative.Pancreas atrophy was observed in one of them.All of the patients received treatment of insulin.