Triptolide （TP）， the core active component of the traditional Chinese medicine Tripterygium wilfordii ， exhibits remarkable pharmacological activities including anti-inflammatory， immunosuppressive and anti-tumor effects， and holds broad application prospects in the treatment of major diseases such as autoimmune diseases and malignant tumors. However， TP has a narrow therapeutic window and causes multi-organ toxicities including liver， kidney and reproductive toxicities， which severely restrict its safe clinical application and new drug development. Therefore， toxicity reduction and efficacy enhancement has become a core scientific problem urgently to be solved in this field. This paper systematically reviews the four core strategies for TP toxicity reduction and efficacy enhancement， including structural modification， dosage form improvement， herbal compatibility， and external therapies of traditional Chinese medicine. Among them， structural modification optimizes the toxic and efficacy characteristics of TP from the molecular structure level， with typica l derivatives including （5 R ）-5-hydroxy triptolide， ZT01， PG490-88， etc. Dosage form modification achieves toxicity reduction and efficacy enhancement via targeted and sustained-controlled drug release of diverse delivery systems. It includes triptolide preparations such as nanoparticles， liposomes， microemulsion gels and liquid crystals， possessing favorable clinical transformation potential. The herbal compatibility and external therapies of traditional Chinese medicine conform to the holistic view of traditional Chinese medicine and have a profound clinical application foundation， but their mechanisms of action are insufficiently elucidated， and they lack unified standardized specifications and high-quality evidence-based proof. In the future， we should rely on multi-omics technology to elucidate the toxic and efficacy mechanisms， integrate technologies to optimize preparations， improve the evaluation system and promote clinical transformation.

Background/Aims: Early detection and effective prognosis prediction in patients with hepatocellular carcinoma (HCC) provide an avenue for survival improvement, yet more effective approaches are greatly needed. We sought to develop the detection and prognosis models with ultra-sensitivity and low cost based on fragmentomic features of circulating cell free mtDNA (ccf-mtDNA). Methods: Capture-based mtDNA sequencing was carried out in plasma cell-free DNA samples from 1168 participants, including 571 patients with HCC, 301 patients with chronic hepatitis B or liver cirrhosis (CHB/LC) and 296 healthy controls (HC). Results: The systematic analysis revealed significantly aberrant fragmentomic features of ccf-mtDNA in HCC group when compared with CHB/LC and HC groups. Moreover, we constructed a random forest algorithm-based HCC detection model by utilizing ccf-mtDNA fragmentomic features. Both internal and two external validation cohorts demonstrated the excellent capacity of our model in distinguishing early HCC patients from HC and highrisk population with CHB/LC, with AUC exceeding 0.983 and 0.981, sensitivity over 89.6% and 89.61%, and specificity over 98.20% and 95.00%, respectively, greatly surpassing the performance of alpha-fetoprotein (AFP) and mtDNA copy number. We also developed an HCC prognosis prediction model by LASSO-Cox regression to select 20 fragmentomic features, which exhibited exceptional ability in predicting 1-year, 2-year and 3-year survival (AUC=0.8333, 0.8145 and 0.7958 for validation cohort, respectively). Conclusions We have developed and validated a high-performing and low-cost approach in a large clinical cohort based on aberrant ccf-mtDNA fragmentomic features with promising clinical translational application for the early detection and prognosis prediction of HCC patients.

Objective To detect the level of anti-drug antibodies(AD As) against anti-tetanus toxin monoclonal antibodies in rhesus monkeys by bridging ELISA and affinity capture elution ELISA(ACE ELISA),and compare them in order to determine an efficient and rapid method for the detection of ADA.Methods The ADA level of anti-tetanus toxin monoclonal antibodies was detected by bridging ELISA and ACE ELISA,and the serum minimum dilution multiple,screening cut point(SCP) and confirmation cut point(CCP) were determined.The sensitivity,drug resistance and precision of the two methods were verified.Forty rhesus monkeys(half male and half female) were randomly divided into negative control group,low dose group,high dose group and intravenous group.The negative control group was inoculated with normal saline,while the other groups were given different doses of anti-tetanus toxin monoclonal antibodies.The blood samples were collected from limbs veins before administration,14 and 28 d after the initial administration and at the end of the recovery period,and the serum was separated,which was detected for the ADA by two methods.Results The minimum dilution multiple of serum was determined to be 1:10.The SCPs of bridging ELISA and ACE ELISA were 0.059 3 and 0.098 1,and the CCPs were 14% and 30%,respectively.The sensitivity of both bridging ELISA and ACE ELISA was 500 ng/mL,but the drug resistance of ACE ELISA at high concentration of positive control antibody(300 μg/mL) was better than that of bridging ELISA(480 ng/mL).The screening precision of bridging ELISA(<5%) was better than that of ACE ELISA(12.35%-21.36%).Four positive samples were detected in rhesus monkey serum samples by ACE ELISA,and three positive samples were confirmed;Eight positive samples were screened by bridging ELISA,while only one was positive after confirmation.The false positive rate of ACE ELISA(0.7%)was lower than that of bridging ELISA(5.2%).Conclusion ACE ELISA has high drug resistance in detecting anti-tetanus toxin monoclonal antibody ADA,which is superior to bridging ELISA,especially in the case of high drug concentration.Although ACE ELISA has slightly lower precision,it is still a reliable ADA detection method,suitable for preclinical research and clinical application of anti-tetanus toxin monoclonal antibodies

Background/Aims: Early detection and effective prognosis prediction in patients with hepatocellular carcinoma (HCC) provide an avenue for survival improvement, yet more effective approaches are greatly needed. We sought to develop the detection and prognosis models with ultra-sensitivity and low cost based on fragmentomic features of circulating cell free mtDNA (ccf-mtDNA). Methods: Capture-based mtDNA sequencing was carried out in plasma cell-free DNA samples from 1168 participants, including 571 patients with HCC, 301 patients with chronic hepatitis B or liver cirrhosis (CHB/LC) and 296 healthy controls (HC). Results: The systematic analysis revealed significantly aberrant fragmentomic features of ccf-mtDNA in HCC group when compared with CHB/LC and HC groups. Moreover, we constructed a random forest algorithm-based HCC detection model by utilizing ccf-mtDNA fragmentomic features. Both internal and two external validation cohorts demonstrated the excellent capacity of our model in distinguishing early HCC patients from HC and highrisk population with CHB/LC, with AUC exceeding 0.983 and 0.981, sensitivity over 89.6% and 89.61%, and specificity over 98.20% and 95.00%, respectively, greatly surpassing the performance of alpha-fetoprotein (AFP) and mtDNA copy number. We also developed an HCC prognosis prediction model by LASSO-Cox regression to select 20 fragmentomic features, which exhibited exceptional ability in predicting 1-year, 2-year and 3-year survival (AUC=0.8333, 0.8145 and 0.7958 for validation cohort, respectively). Conclusions We have developed and validated a high-performing and low-cost approach in a large clinical cohort based on aberrant ccf-mtDNA fragmentomic features with promising clinical translational application for the early detection and prognosis prediction of HCC patients.

Background/Aims: Early detection and effective prognosis prediction in patients with hepatocellular carcinoma (HCC) provide an avenue for survival improvement, yet more effective approaches are greatly needed. We sought to develop the detection and prognosis models with ultra-sensitivity and low cost based on fragmentomic features of circulating cell free mtDNA (ccf-mtDNA). Methods: Capture-based mtDNA sequencing was carried out in plasma cell-free DNA samples from 1168 participants, including 571 patients with HCC, 301 patients with chronic hepatitis B or liver cirrhosis (CHB/LC) and 296 healthy controls (HC). Results: The systematic analysis revealed significantly aberrant fragmentomic features of ccf-mtDNA in HCC group when compared with CHB/LC and HC groups. Moreover, we constructed a random forest algorithm-based HCC detection model by utilizing ccf-mtDNA fragmentomic features. Both internal and two external validation cohorts demonstrated the excellent capacity of our model in distinguishing early HCC patients from HC and highrisk population with CHB/LC, with AUC exceeding 0.983 and 0.981, sensitivity over 89.6% and 89.61%, and specificity over 98.20% and 95.00%, respectively, greatly surpassing the performance of alpha-fetoprotein (AFP) and mtDNA copy number. We also developed an HCC prognosis prediction model by LASSO-Cox regression to select 20 fragmentomic features, which exhibited exceptional ability in predicting 1-year, 2-year and 3-year survival (AUC=0.8333, 0.8145 and 0.7958 for validation cohort, respectively). Conclusions We have developed and validated a high-performing and low-cost approach in a large clinical cohort based on aberrant ccf-mtDNA fragmentomic features with promising clinical translational application for the early detection and prognosis prediction of HCC patients.

Ovulatory disorders are mainly characterized by abnormal follicular maturation or ovulation， with complex etiologies and a lack of effective prevention and treatment methods. Autophagy dysfunction is closely related to the generation and progression of ovulatory disorders. This article systematically elucidates the mechanisms of TCM on follicular development and ovulatory disorders from the perspective of autophagy. It also reviews relevant studies on how TCM regulates autophagy to influence follicular development and improve ovulatory disorders. The findings reveal that TCM monomers/active ingredients （leonurine， total flavonoids from Eucommia ulmoides， alpinetin， icariin， etc.） and compound formulas （including Cangfu daotan decoction， Guishen yugong decoction， Zhuluan decoction， Yishen yangluan formula， Guishen pill， etc.） improve the follicular microenvironment， regulate sex hormone levels， and reduce follicular atresia by regulating autophagy-related genes and signaling pathways such as phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin and AMP-activated protein kinase. These actions thereby promote normal follicular development and ovulation， and delay ovarian aging. Most research in this field is based on cellular and animal experiments， often focusing on a single signaling pathway or factor. Some studies fail to fully reflect the individualized treatment characteristics of TCM that emphasize “syndrome differentiation and treatment”， highlighting the urgent need for further investigation.

OBJECTIVES: To evaluate the strategies and outcomes of assisted reproductive technology (ART) for fertility preservation and promotion in women with malignant tumors, and to analyze ART outcomes across different tumor types. METHODS: We conducted a retrospective analysis of female patients who underwent ART for fertility preservation or treatment at the Reproductive Center of the Women's Hospital, Zhejiang University School of Medicine, between January 1, 2018, and December 31, 2023. A total of 163 ART-aided pregnancy patients with malignant tumors were included in the case group, among which 6 patients underwent embryo cryopreservation for fertility preservation before radiotherapy or chemotherapy. Additionally, 11 unmarried women underwent oocyte cryopreservation due to borderline ovarian tumors, ovarian cancer, breast cancer, or hematological malignancies. The control group was selected from women without a history of malignant tumors who received ART treatment during the same period, using propensity score matching at a ratio of 1∶2, resulting in 326 cases. Data were collected through the reproductive medical record system and telephone follow-up (as of October 31, 2024). Baseline characteristics, controlled ovarian hyperstimulation parameters, laboratory indicators, and pregnancy outcomes were compared between case and control groups and among patients with different tumor types, and the tumor recurrence of the patients was followed up. RESULTS: Patients in the case group had significantly lower ovarian reserve (AMH, AFC) and a higher proportion of diminished ovarian reserve compared to the control group (all P<0.01). Regarding the ovulation induction protocol, the proportion of patients using the minimal stimulation protocol in the case group was significantly higher than that in the control group (29.45% vs. 12.88%, P<0.01), and the total dosage of gonadotropins used was lower (P<0.01). In terms of assisted reproductive outcomes, there were no statistically significant differences between the two groups in the number of retrieved oocytes, number of high-quality embryos, fertilization rate, cumulative pregnancy rate, cumulative live birth rate, or miscarriage rate (all P>0.05). However, the number of oocyte retrieval cycles and embryo transfer cycles required to achieve a live birth outcome in the case group were significantly higher than those in the control group (both P<0.05). Subgroup analysis showed that there were no significant differences in cumulative pregnancy rate and live birth rate among patients with different tumor types (thyroid cancer, reproductive system tumors, breast cancer, lung cancer). Nevertheless, lung cancer patients had the lowest ovarian reserve and required the most oocyte retrieval cycles due to their older age; breast cancer patients had a relatively lower fertilization rate partially because some of them were complicated with male factors. A follow-up of 154 tumor patients (with a follow-up rate of 88.5%) revealed that 6 patients (4.20%) had tumor recurrence, and 1 breast cancer patient died due to tumor recurrence. None of the 11 unmarried patients who had undergone oocyte cryopreservation had used the cryopreserved oocytes for assisted pregnancy yet, and 1 patient who had undergone fertility preservation died due to tumor recurrence. CONCLUSIONS Women of reproductive age with malignant tumors are at risk of diminished fertility. ART can effectively preserve and promote fertility, enabling favorable pregnancy and live birth outcomes. It is recommended to initiate a multidisciplinary assessment promptly prior to radiotherapy/chemotherapy and formulate an individualized ART regimen for fertility preservation or promotion, so as to achieve reproductive goals or safeguard future fertility potential.

Objective:To analyze effects of T11TS in the treatment of Cryptococcus neoformans fungal infection and the regula-tion of T-cell calcineurin(CaN)-activated T lymphoid nuclear factor(NFAT)pathway.Methods:SD rats infected with Cryptococcus neoformans were treated with immunoenhancer T11TS.Flow cytometry,Western blot and nuclear translocation were used to detect the changes of T cell function and related signal pathway.Results:The number of Cryptococcus neoformans in lung and spleen of CT3 group was significantly less than Cryptococcus neoformans infection group,CT1 group and CT2 group(P<0.05).The results of flow cy-tometry showed that the expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 in Cryptococcus neoformans infection group were significantly lower than control group(P<0.05).The expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 of CT1,CT2 and CT3 groups were significantly higher than Cryptococcus neoformans infection group(P<0.05).The results of Western blot were similar to those of flow cytometry.Conclusion:T11TS in the treatment of Cryptococcus neoformans infection can enhance the immunity of the body by up-regulating the expression of related signal factors in the CaN-NFAT pathway of T lymphocytes,increasing the expression of NFAT and IL-2,stimulating the activation and proliferation of T lymphocytes.

Objective:To analyze effects of T11TS in the treatment of Cryptococcus neoformans fungal infection and the regula-tion of T-cell calcineurin(CaN)-activated T lymphoid nuclear factor(NFAT)pathway.Methods:SD rats infected with Cryptococcus neoformans were treated with immunoenhancer T11TS.Flow cytometry,Western blot and nuclear translocation were used to detect the changes of T cell function and related signal pathway.Results:The number of Cryptococcus neoformans in lung and spleen of CT3 group was significantly less than Cryptococcus neoformans infection group,CT1 group and CT2 group(P<0.05).The results of flow cy-tometry showed that the expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 in Cryptococcus neoformans infection group were significantly lower than control group(P<0.05).The expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 of CT1,CT2 and CT3 groups were significantly higher than Cryptococcus neoformans infection group(P<0.05).The results of Western blot were similar to those of flow cytometry.Conclusion:T11TS in the treatment of Cryptococcus neoformans infection can enhance the immunity of the body by up-regulating the expression of related signal factors in the CaN-NFAT pathway of T lymphocytes,increasing the expression of NFAT and IL-2,stimulating the activation and proliferation of T lymphocytes.

OBJECTIVE To evaluate the quality of diagnosis and treatment guidelines and expert consensuses on childhood immune thrombocytopenic purpura(ITP)published domestically and internationally,in order to provide reference for clinical practice and future guideline/expert consensus development and improvement.METHODS A systematic search was conducted across multiple databases,including PubMed,Cochrane Library,Embase,CNKI,Wanfang data,VIP,CBM;additionally,supplementary searches were carried out on websites such as Medlive,the Chinese Medical Association's official website,and National Institute for Health and Clinical Excellence in the UK.The retrieval time ranged from the inception to September 2,2024.Researchers who had undergone systematic training independently evaluated the methodology and report quality included in the guideline/consensus using the Appraisal of Guidelines Research and Evaluation Ⅱ(AGREE Ⅱ)and the Reporting Items for Practice Guidelines in Healthcare(RIGHT).RESULTS A total of 11 guidelines/consensuses were included.The average scores for the six domains of AGREE Ⅱ tool respectively were"range and purpose"[(66.67±17.98)%],"participants"[58.33%(13.89%,73.61%)],"rigor"[(41.81±23.85)%],"clarity"[(69.57±19.35)%],"applicability"[(35.98±17.83)%],and"independence"[27.08%(0,75.00%)];out of 11 articles,9 had a recommendation level of B,2 had a recommendation level of C,and there were no A-level articles.The average reporting rates of the 7 areas in the RIGHT tool were"basic information"[(72.35±12.95)%],"background"[(54.55±15.40)%],"evidence"[(36.36±24.81)%],"recommended opinions"[(53.25±19.20)%],"review and quality assurance"[0(0,25.00%)],"funding and conflict of interest statement and management"[12.50%(0,25.00%)],and other aspects[8.33%(0,50.00%)].In addition,there was no statistically significant difference in the AGREE Ⅱ and RIGHT scores between the guidelines and consensuses(P＞0.05).CONCLUSIONS The overall quality of the guidelines and consensuses included in this study is not high,with a recommended level of B or C.It is recommended that clinical decision-making prioritize referring to the relatively high-quality guideline/consensus among them.The quality of evidence in the existing traditional Chinese medicine guidelines for children with ITP needs to be improved,and there is no integrated guideline/consensus for traditional Chinese and Western medicine.It is recommended to revise or write relevant guideline/consensus according to the requirements of AGREE Ⅱ and RIGHT in various fields to guide clinical practice.