Tacrolimus is an immunosuppressant that was clinically used for organ transplantation in the 1990s. In the early 2000s, tacrolimus began to be used to treat pediatric kidney diseases in China. This article reviews the therapeutic effects, clinical dosages, and treatment methods of tacrolimus in the treatment of steroid-resistant, steroid-dependent, frequently relapsing, different pathological types, and monogenic mutation-related childhood nephrotic syndrome. It explores the clinical guiding role of machine learning in tacrolimus treatment for childhood nephrotic syndrome, aiming to provide references for the clinical research and application of tacrolimus in pediatric kidney diseases.

Objective:To investigate the molecular pathogenic mechanism of venous thrombosis caused by heterozygous missense mutations in two protein C (PROC) genes through laboratory phenotype analysis, genetic mutation analysis, and in vitro expression experiments.Methods:Two probands presented with venous thromboembolism at the First Affiliated Hospital of Wenzhou Medical University. Clinical data and blood samples were collected from the probands and their family members to evaluate the plasma protein C (PC) activity (PC∶A), PC antigen (PC∶Ag) levels, and other relevant coagulation parameters. The anticoagulant capacity was assessed using the thrombin generation test (TGT). The mutation sites of the PROC gene were identified using direct DNA sequencing. Bioinformatics software was used to analyze the conservation and pathogenicity of the mutated gene. PyMOL software was used for the analysis of the protein three-dimensional models and interactions between mutated amino acids. Wild-type and two mutant expression vectors were constructed and HEK293T cells were transiently transfected. Total cellular RNA was extracted from positively transfected cells to investigate the transcriptional levels of the mutant PROC gene. Enzyme-linked immunosorbent assay, Western blot, and cellular immunofluorescence assays were used to investigate the translation levels of the mutant PROC protein.Results:Probands 1 and 2 exhibited PC∶A levels of 35% and 40% and PC∶Ag levels of 44% and 39%, with increasing D-dimer levels to 4.42 mg/L and 0.83 mg/L, respectively. Meanwhile, other coagulation parameters revealed no significant abnormalities. TGT demonstrated impaired anticoagulant function in both proband witnesses and their familial PC carriers. Sequencing analysis revealed heterozygous missense mutations c. 833T>C (p. Leu278Pro) in proband 1 and c. 1330T>C (p. Trp444Arg) in proband 2 within exon 9 of the PROC gene. Conservation analysis revealed that Leu278 and Trp444 were highly conserved across homologous species. Pathogenicity analysis indicated that both p. Leu278Pro and p. Trp444Arg mutations are deleterious. Protein modeling analysis demonstrated that both mutations induce structural alterations in the protein. In vitro expression experiments revealed that compared with the wild-type, both p. Leu278Pro and p. Trp444Arg mutations showed no significant differences in the mRNA expression level of the PC protein. However, both mutations caused significantly lower PC∶Ag content and protein expression levels in the cell culture supernatant compared with the wild-type, whereas higher levels were observed in the cell culture lysate. This indicates the association of both mutations with the secretion function of the PC protein.Conclusion:The heterozygous missense mutations p. Leu278Pro and p. Trp444Arg in exon 9 of the PROC gene in both probands are associated with decreased PC levels.

[Objective] To validate and optimize the platelet transfusion refractoriness (PTR) prediction model for patients with hematological disorders established by our center. [Methods] The data of patients with hematological diseases who received platelet transfusions from December 2021 to December 2022 were used as the training set, and data from January 2023 to December 2023 as the validation set. The validation set data was used to validate the predictive model constructed on the training set. Relevant risk factors for PTR were collected through literature review and preliminary studies。 The patients were divided into effective and ineffective groups according to the corrected count increment (CCI) of platelet counts. Predictive factors were screened using univariate and multivariate logistic regression. The calibration of the model were assessed via calibration curves, while discrimination, accuracy, sensitivity, and specificity were evaluated using receiver operating characteristic (ROC) curves Clinical utility was further analyzed with decision curve analysis (DCA). [Results] The Hosmer-Lemeshow (H-L) goodness-of-fit test for the validation set yielded S: P=0.000, indicating that the original model needs optimization. Baseline comparisons and logistic regression identified the number of red blood cell units (RBCU) and platelet units (PLT-U) transfused as key predictors for the optimized model. The H-L goodness-of-fit test S: P values for the training and validation sets were 0.930 and 0.056, respectively; the ROC areas were 0.793 5 and 0.809 4, specificities 90.95% and 84.21%, sensitivities 59.26% and 70.04%, and accuracies 78.14% and 74.10%, respectively. DCA demonstrated clinical net benefit within a prediction probability threshold range of 0.2-0.8. [Conclusion] Transfusion volumes of RBC-U and PLT-U were inversely associated with PTR in hematological patients. The resulting PTR prediction model exhibits moderate predictive efficacy and clinical benefit.

Hyperuricemia is a chronic metabolic disease resulting from purine metabolic dysfunction and is classified under the category of"blood turbidity"in traditional Chinese medicine.Our team termed it"acid turbidity,"and its pathogenesis is closely related to the dynamic evolution of the clear and the turbid components.With the change of modern people's diet structure,the incidence of hyperuricemia is increasing annually owing to the intake of fatty,sweet foods and alcohol.Therefore,this paper explores hyperuricemia from the"indigestion of spleen and stomach"theory.The core pathogenesis of hyperuricemia is indigestion of spleen and stomach,the inversion of clear and turbid substances,and endogenous acid turbidity.The initial manifestation of hyperuricemia is the internal retention of acid turbidity and ascending-descending disharmony;the gradual manifestation of this disease is that indigestion causes heat,and acid turbidity transforms into poison;the final manifestation of this disease is that secular indigestion causes deficiency and the inversion of clear and turbid substances.It can be summarized into three syndromes:syndromes of internal retention of dampness-turbidity,dampness-heat toxin amassment,and dampness-heat due to spleen deficiency.Therefore,this paper proposes to treat the disease according to different syndromes,with ascending the clear and descending the turbid as the core of treatment.And the therapeutic approach employs the flexible application of three methods:transportation,resolving,and transformation.For syndrome of internal retention of dampness-turbidity,treatment focuses on promoting spleen transportation to eliminate dampness;for syndrome of dampness-heat toxin amassment,the strategy is to resolve indigestion and purge heat;and for syndrome of dampness-heat due to spleen deficiency,the aim is to resolve turbidity and clear heat.By ascending the clear and descending the turbid,so that"returning the clear and the turbid to the original,"the spleen and stomach regain harmony,functions of ascending and descending are reestablished,and hyperuricemia can be effectively managed.

Although fertility-preserving treatment strategies have demonstrated significant clinical efficacy in patients with early-stage endometrial cancer(EC)and atypical endometrial hyperplasia(AEH),some patients who experience recurrence still express a strong desire for fertility and request conservative retreatment.This poses new challenges for clinical management.This article reviews the latest research advances in retreatment for recurrent patients and synthesizes findings from previous studies to draw the following conclusions.First,before formulating a retreatment plan,it is crucial to comprehensively evaluate key factors such as tumor characteristics,fertility intentions,and overall health status of the patient.These factors collectively determine the feasibility and appropri-ateness of an individualized retreatment strategy.Second,studies have shown that retreatment options for AEH/EC are diverse,with high-dose oral progestin remaining the primary approach.Combining progestin with metformin,GnRH-a,and hysteroscopic resection can enhance treatment outcomes.Individualized treatment plans should be tailored to the specific conditions of each patient.Based on literature analysis,the complete remission(CR)rate of retreatment ranges from 81.1%to 88.6%,with a pregnancy rate of 26.5%-50.0%and a live birth rate of 14.3%-29.0%.However,the recurrence rate remains high at 24.5%-45.5%,significantly higher than that of initial treatment.This indicates that while retreatment has achieved some success in disease control and fertility preservation,the risk of recurrence still requires significant attention.Therefore,a strict monitoring and follow-up system must be established during retreatment.Additionally,studies have identified factors associated with a higher risk of recurrence,including age over 35,overweight or obesity,polycystic ovarian syndrome,metabolic syndrome,high pathological grade,and advanced clinical stage.On the other hand,pregnancy following CR,weight loss,and maintenance therapy serve as protective factors against disease recurrence.Patients are encouraged to actively engage in weight management during treatment and consider initiating assisted reproductive technology promptly after achieving CR to optimize pregnancy outcomes while minimizing the risk of recurrence.Future research should prioritize investigating precision treatment strategies informed by molecular classification and identifying predictive biomarkers,thereby enabling the development of more personalized and precise treatment plans tailored to indi-vidual patients.

Although fertility-preserving treatment strategies have demonstrated significant clinical efficacy in patients with early-stage endometrial cancer(EC)and atypical endometrial hyperplasia(AEH),some patients who experience recurrence still express a strong desire for fertility and request conservative retreatment.This poses new challenges for clinical management.This article reviews the latest research advances in retreatment for recurrent patients and synthesizes findings from previous studies to draw the following conclusions.First,before formulating a retreatment plan,it is crucial to comprehensively evaluate key factors such as tumor characteristics,fertility intentions,and overall health status of the patient.These factors collectively determine the feasibility and appropri-ateness of an individualized retreatment strategy.Second,studies have shown that retreatment options for AEH/EC are diverse,with high-dose oral progestin remaining the primary approach.Combining progestin with metformin,GnRH-a,and hysteroscopic resection can enhance treatment outcomes.Individualized treatment plans should be tailored to the specific conditions of each patient.Based on literature analysis,the complete remission(CR)rate of retreatment ranges from 81.1%to 88.6%,with a pregnancy rate of 26.5%-50.0%and a live birth rate of 14.3%-29.0%.However,the recurrence rate remains high at 24.5%-45.5%,significantly higher than that of initial treatment.This indicates that while retreatment has achieved some success in disease control and fertility preservation,the risk of recurrence still requires significant attention.Therefore,a strict monitoring and follow-up system must be established during retreatment.Additionally,studies have identified factors associated with a higher risk of recurrence,including age over 35,overweight or obesity,polycystic ovarian syndrome,metabolic syndrome,high pathological grade,and advanced clinical stage.On the other hand,pregnancy following CR,weight loss,and maintenance therapy serve as protective factors against disease recurrence.Patients are encouraged to actively engage in weight management during treatment and consider initiating assisted reproductive technology promptly after achieving CR to optimize pregnancy outcomes while minimizing the risk of recurrence.Future research should prioritize investigating precision treatment strategies informed by molecular classification and identifying predictive biomarkers,thereby enabling the development of more personalized and precise treatment plans tailored to indi-vidual patients.

Hyperuricemia is a chronic metabolic disease resulting from purine metabolic dysfunction and is classified under the category of"blood turbidity"in traditional Chinese medicine.Our team termed it"acid turbidity,"and its pathogenesis is closely related to the dynamic evolution of the clear and the turbid components.With the change of modern people's diet structure,the incidence of hyperuricemia is increasing annually owing to the intake of fatty,sweet foods and alcohol.Therefore,this paper explores hyperuricemia from the"indigestion of spleen and stomach"theory.The core pathogenesis of hyperuricemia is indigestion of spleen and stomach,the inversion of clear and turbid substances,and endogenous acid turbidity.The initial manifestation of hyperuricemia is the internal retention of acid turbidity and ascending-descending disharmony;the gradual manifestation of this disease is that indigestion causes heat,and acid turbidity transforms into poison;the final manifestation of this disease is that secular indigestion causes deficiency and the inversion of clear and turbid substances.It can be summarized into three syndromes:syndromes of internal retention of dampness-turbidity,dampness-heat toxin amassment,and dampness-heat due to spleen deficiency.Therefore,this paper proposes to treat the disease according to different syndromes,with ascending the clear and descending the turbid as the core of treatment.And the therapeutic approach employs the flexible application of three methods:transportation,resolving,and transformation.For syndrome of internal retention of dampness-turbidity,treatment focuses on promoting spleen transportation to eliminate dampness;for syndrome of dampness-heat toxin amassment,the strategy is to resolve indigestion and purge heat;and for syndrome of dampness-heat due to spleen deficiency,the aim is to resolve turbidity and clear heat.By ascending the clear and descending the turbid,so that"returning the clear and the turbid to the original,"the spleen and stomach regain harmony,functions of ascending and descending are reestablished,and hyperuricemia can be effectively managed.

Objective:To investigate the molecular pathogenic mechanism of venous thrombosis caused by heterozygous missense mutations in two protein C (PROC) genes through laboratory phenotype analysis, genetic mutation analysis, and in vitro expression experiments.Methods:Two probands presented with venous thromboembolism at the First Affiliated Hospital of Wenzhou Medical University. Clinical data and blood samples were collected from the probands and their family members to evaluate the plasma protein C (PC) activity (PC∶A), PC antigen (PC∶Ag) levels, and other relevant coagulation parameters. The anticoagulant capacity was assessed using the thrombin generation test (TGT). The mutation sites of the PROC gene were identified using direct DNA sequencing. Bioinformatics software was used to analyze the conservation and pathogenicity of the mutated gene. PyMOL software was used for the analysis of the protein three-dimensional models and interactions between mutated amino acids. Wild-type and two mutant expression vectors were constructed and HEK293T cells were transiently transfected. Total cellular RNA was extracted from positively transfected cells to investigate the transcriptional levels of the mutant PROC gene. Enzyme-linked immunosorbent assay, Western blot, and cellular immunofluorescence assays were used to investigate the translation levels of the mutant PROC protein.Results:Probands 1 and 2 exhibited PC∶A levels of 35% and 40% and PC∶Ag levels of 44% and 39%, with increasing D-dimer levels to 4.42 mg/L and 0.83 mg/L, respectively. Meanwhile, other coagulation parameters revealed no significant abnormalities. TGT demonstrated impaired anticoagulant function in both proband witnesses and their familial PC carriers. Sequencing analysis revealed heterozygous missense mutations c. 833T>C (p. Leu278Pro) in proband 1 and c. 1330T>C (p. Trp444Arg) in proband 2 within exon 9 of the PROC gene. Conservation analysis revealed that Leu278 and Trp444 were highly conserved across homologous species. Pathogenicity analysis indicated that both p. Leu278Pro and p. Trp444Arg mutations are deleterious. Protein modeling analysis demonstrated that both mutations induce structural alterations in the protein. In vitro expression experiments revealed that compared with the wild-type, both p. Leu278Pro and p. Trp444Arg mutations showed no significant differences in the mRNA expression level of the PC protein. However, both mutations caused significantly lower PC∶Ag content and protein expression levels in the cell culture supernatant compared with the wild-type, whereas higher levels were observed in the cell culture lysate. This indicates the association of both mutations with the secretion function of the PC protein.Conclusion:The heterozygous missense mutations p. Leu278Pro and p. Trp444Arg in exon 9 of the PROC gene in both probands are associated with decreased PC levels.

Tacrolimus is an immunosuppressant that was clinically used for organ transplantation in the 1990s. In the early 2000s, tacrolimus began to be used to treat pediatric kidney diseases in China. This article reviews the therapeutic effects, clinical dosages, and treatment methods of tacrolimus in the treatment of steroid-resistant, steroid-dependent, frequently relapsing, different pathological types, and monogenic mutation-related childhood nephrotic syndrome. It explores the clinical guiding role of machine learning in tacrolimus treatment for childhood nephrotic syndrome, aiming to provide references for the clinical research and application of tacrolimus in pediatric kidney diseases.

Objective To analyze and identify the volatile constituents in different parts(flowers,stems and leaves)of Huai chrysanthemumin,and to lay a theoretical foundation for the comprehensive utilization for it.Methods The volatile oil in different parts of Huai chrysanthemumin were extracted by hydrodistillation,respectively.Their constituents were analyzed by gas chromatography-mass spectrometry(GC-MS).The compounds were identified by library search and literature screening.The relative percentage of each compound was obtained by the area normalization method.The differences in their chemical compositions were analyzed by Venn diagram,principal component analysis(PCA)and cluster heat map analysis.Results A total of 62 volatile chemical components were identified from different parts of Huai chrysanthemumin,including monoterpenes,sesquiterpenes,and their derivatives,as well as a small amount of aliphatic compounds.32,42 and 40 volatile components were detected from the flowers,stems and flowers,respectively.Furthermore 17 volatile components were shared by three parts,whereas 5,6 and 16 volatile components were unique to the flowers,stems and leaves,respectively.The results of stoichiometric analysis showed that both PCA and cluster heat map analysis could separate the flowers,stems and leaves,and their volatile components were different.Conclusion The types and contents of the volatile oil in the stems,leaves and flowers of Huai chrysanthemumin have certain variability,which provide a scientific basis for the further medicinal or industrial exploitation of different parts of Huai chrysanthemumin.