ObjectiveGastrodia elata has evolved ecological types with shortened rhizome internodes and diversified flower and fruit coloration in response to different altitudes. Studying the genetic mechanisms of different ecotype germplasm is significant for guiding variety breeding in different cultivation areas. MethodsThe bHLH gene family was identified based on the whole-genome datasets of G. elata f. elata and G. elata f. glauca. Subsequently， the gene family members were subject to analysis， including gene structure， chromosomal localization， cis-acting elements， gene synteny， and phylogeny. Combined with transcriptome data and quantitative Real-time PCR， the expression patterns of bHLH genes in the stems of the different G. elata ecotype germplasm were analyzed. Finally， correlation analysis was conducted between gene expression patterns and color to obtain the key bHLH genes regulating the color formation of stem. ResultsA total of 63 bHLH genes were identified in both G elata f. elata and G. elata f. glauca， unevenly distributed across 17 chromosomes and clustered into 16 subfamilies， with significant expansion in some family members. Obvious inversions of bHLH genes on the same chromosome and interchromosomal translocations were detected in the two ecotype germplasm. Among these genes， 12 bHLH genes （such as bHLH62-3 and bHLH74） were associated with the bright yellow color of G elata f. elata stem， while 9 bHLH genes （such as PIL13， UNE12， and bHLH130） were correlated with the red color of G. elata f. glauca stem. Compared to G. elata f. glauca， the bHLH48 expression level was significantly higher in flowers and scale leaves of G elata f. elata， and the bHLH62-3 expression level was significantly higher in all organs of G elata f. elata. ConclusionsFunctional pathway divergence of the bHLH family members has occurred across different chromosomes in G elata f. elata and G. elata f. glauca. Through synergism or antagonism with other genes， 21 bHLH genes participate in the coloration metabolic pathway regulation of stems， flowers， and fruits. Specifically， bHLH62-3 is involved in regulating stem color differentiation in the anthocyanin biosynthesis pathway of G. elata， thus relevant to the color formation of stem. Additionally， GebHLH48 positively regulates flowering-related pathways to promote the early-flowering phenotype of G. elata f. elata. These findings have laid the foundation for analyzing the genetic regulatory mechanisms underlying the color formation of the G. elata stem.

ObjectiveGastrodia elata has evolved ecological types with shortened rhizome internodes and diversified flower and fruit coloration in response to different altitudes. Studying the genetic mechanisms of different ecotype germplasm is significant for guiding variety breeding in different cultivation areas. MethodsThe bHLH gene family was identified based on the whole-genome datasets of G. elata f. elata and G. elata f. glauca. Subsequently， the gene family members were subject to analysis， including gene structure， chromosomal localization， cis-acting elements， gene synteny， and phylogeny. Combined with transcriptome data and quantitative Real-time PCR， the expression patterns of bHLH genes in the stems of the different G. elata ecotype germplasm were analyzed. Finally， correlation analysis was conducted between gene expression patterns and color to obtain the key bHLH genes regulating the color formation of stem. ResultsA total of 63 bHLH genes were identified in both G elata f. elata and G. elata f. glauca， unevenly distributed across 17 chromosomes and clustered into 16 subfamilies， with significant expansion in some family members. Obvious inversions of bHLH genes on the same chromosome and interchromosomal translocations were detected in the two ecotype germplasm. Among these genes， 12 bHLH genes （such as bHLH62-3 and bHLH74） were associated with the bright yellow color of G elata f. elata stem， while 9 bHLH genes （such as PIL13， UNE12， and bHLH130） were correlated with the red color of G. elata f. glauca stem. Compared to G. elata f. glauca， the bHLH48 expression level was significantly higher in flowers and scale leaves of G elata f. elata， and the bHLH62-3 expression level was significantly higher in all organs of G elata f. elata. ConclusionsFunctional pathway divergence of the bHLH family members has occurred across different chromosomes in G elata f. elata and G. elata f. glauca. Through synergism or antagonism with other genes， 21 bHLH genes participate in the coloration metabolic pathway regulation of stems， flowers， and fruits. Specifically， bHLH62-3 is involved in regulating stem color differentiation in the anthocyanin biosynthesis pathway of G. elata， thus relevant to the color formation of stem. Additionally， GebHLH48 positively regulates flowering-related pathways to promote the early-flowering phenotype of G. elata f. elata. These findings have laid the foundation for analyzing the genetic regulatory mechanisms underlying the color formation of the G. elata stem.

Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

Aim To evaluate the toxicity and mecha-nism of Epimedium sagittatum(Sieb.et Zucc.)Maxim aqueous extract(ESMAE)on HepaRG cells based on serum toxicology.Methods MTT assay was used to detect the activity of HepaRG cells after treatment with the serum containing ESMAE from SD rats.Western blot was used to detect the effects of the serum contai-ning drug on the expression of endoplasmic reticulum stress-related proteins(PERK,eIF-2α,ATF-4,GRP78,CHOP)and pyroptosis related proteins(NL-RP1,caspase-1,cleaved caspase-1,GSDMD,GSDMD-N).MTT assay was used to detect the activity of Hep-aRG cells after treatment with the liver homogenate containing ESMAE from SD rats.Results Twenty percent serum containing drug significantly decreased the viability of HepaRG cells,with the cells exhibiting swelling,rupture,and vesicle-like pyroptosis.The ex-pression levels of endoplasmic reticulum stress-related proteins PERK,eIF-2α,ATF-4,GRP78,and CHOP significantly increased.The expression levels of pro-teins involved in the NLRP1-mediated classical pyrop-tosis pathway were significantly increased.Finally the liver homogenate containing drug decreased the cell ac-tivity,and cells exhibited swelling,rupture,and vesicle-like pyroptosis.Conclusions After administration of ESMAE,the serum containing drug and the liver ho-mogenate containing drug of rats show toxicity to Hep-aRG cells,and can induce endoplasmic reticulum stress and activate the NLRP1/caspase-1/GSDMD pyroptosis pathway in HepaRG cells.

BACKGROUND:The combination of traditional Chinese medicine syndrome and semen quality-related parameters can jointly predict the occurrence of abnormal increase in sperm DNA fragmentation index (DFI) and draw a column chart,which can significantly improve clinical practicality and application efficiency,provide a basis for comprehensive evaluation of semen quality in clinical practice,take active intervention measures to improve clinical outcomes,and formulate personalized medical plans.OBJECTIVE:To explore the prediction model and verification of sperm DNA fragments based on traditional Chinese medicine syndrome and semen quality-related parameters.METHODS:Retrospective analysis was made on 420 infertile patients who received traditional Chinese medicine syndrome diagnosis and sperm DNA fragment rate examination in the Department of Traditional Chinese Medicine Andrology,Nanxishan Hospital of Guangxi Zhuang Autonomous Region from July 2019 to July 2021.According to the Manual of Human Semen Examination and Treatment Laboratories (6th Edition),137 patients with sperm DFI>30％ were included in the group of abnormally high sperm DFI,and 283 patients with sperm DFI ≤ 30％ were taken as the control group.First,univariate analysis was used to screen the influencing factors of the abnormal increase of sperm DFI.Then,the best matching factor was selected by using the collinearity problem of LASSO correction factors.Then,it was included in the multifactor forward stepwise logistic regression to find out its independent influencing factors and draw a nomogram.Finally,the receiver operating characteristic curve,calibration curve,decision curve analysis and clinical impact curve were used to verify the differentiation and accuracy of the prediction model and its clinical application effectiveness.RESULTS AND CONCLUSION:(1) The results of the univariate analysis showed that age,body mass index,forward motion rate,total sperm motility,sperm concentration,sperm morphology,kidney yang deficiency syndrome,damp heat downpour syndrome,and kidney sperm deficiency syndrome were the influencing factors for the abnormal increase of sperm DFI (P<0.05).(2) The best matching factors further screened by LASSO regression were age,body mass index,total sperm motility,sperm concentration,sperm morphology,kidney yang deficiency syndrome,damp heat downpour syndrome,and kidney essence deficiency syndrome (P<0.05).(3) Multifactor forward stepwise Logistic regression showed that age,body mass index,sperm concentration,total sperm motility,damp heat downpour syndrome,and kidney yang deficiency syndrome were six independent factors that caused the abnormal increase in sperm DFI.(4) Receiver operating characteristic curve showed that the area under the curve of the model group was 0.760(0.713,0.806),and the area under the curve of the validation group was 0.745(0.714,0.776).It showed that the prediction model had good discrimination.(5) The average absolute error of the calibration curve was 0.040,and the Hosmer Lemeshow test (P>0.05),suggesting that there was no significant statistical difference between the probability of the abnormal increase in DFI of spermatozoa predicted by the model and the probability of the abnormal increase in DFI of spermatozoa actually occurred,which confirmed that the model had good accuracy.(6) Decision curve analysis and clinical impact curve showed that the model group and validation group had the maximum clinical net benefit when the threshold probability values were (0.08-0.84) and (0.09-0.78) respectively,and had good clinical application efficiency within the threshold probability range.(7) These findings conclude that age,body mass index,sperm concentration,total sperm viability,damp heat downpour syndrome and kidney yang deficiency syndrome are independent factors that cause the abnormal increase in sperm DFI.The nomogram of the clinical prediction model constructed by them has good clinical prediction value and clinical application efficiency,and can provide the basis for comprehensive clinical evaluation of semen quality and individualized medical service.

A 3-month-old female infant was admitted for incessant crying for 3 days.Examination revealed a rapidly growing massive retroperitoneal mass that was difficult to resect.Needle biopsy confirmed infantile fibrosarcoma.Initial chemotherapy with the VAC regimen(vincristine,actinomycin D,and cyclophosphamide)was administered,but the response was poor.The common fusion in infantile fibrosarcoma is ETV6-NTRK3,and next-generation sequencing detected an RBPMS-NTRK3 gene fusion in this patient.To our knowledge,this is the first reported case of infantile fibrosarcoma with RBPMS-NTRK3 fusion in China.Treatment with larotrectinib resulted in marked tumor shrinkage.

A 3-month-old female infant was admitted for incessant crying for 3 days.Examination revealed a rapidly growing massive retroperitoneal mass that was difficult to resect.Needle biopsy confirmed infantile fibrosarcoma.Initial chemotherapy with the VAC regimen(vincristine,actinomycin D,and cyclophosphamide)was administered,but the response was poor.The common fusion in infantile fibrosarcoma is ETV6-NTRK3,and next-generation sequencing detected an RBPMS-NTRK3 gene fusion in this patient.To our knowledge,this is the first reported case of infantile fibrosarcoma with RBPMS-NTRK3 fusion in China.Treatment with larotrectinib resulted in marked tumor shrinkage.

Managing look-alike/sound-alike medications is important to medical institutions'pharmaceutical manage-ment and pharmacy services.Based on national policies and regulations,this standard focuses on the whole life cycle of look-alike/sound-alike medications in medical institutions.It is developed based on the principles of scientific validity,universality,guidance,and applicability,formed by sorting out problems,soliciting opinions,expert argumentation,and deliberation.It is the first group standard to standardize the management of look-alike/sound-alike medications.This paper introduced and analyzed the team com-position,problem sorting and compilation process,and various elements of the standard in the process of formulating the standard,to provide a reference for medical institutions to use this standard.

Aim To evaluate the toxicity and mecha-nism of Epimedium sagittatum(Sieb.et Zucc.)Maxim aqueous extract(ESMAE)on HepaRG cells based on serum toxicology.Methods MTT assay was used to detect the activity of HepaRG cells after treatment with the serum containing ESMAE from SD rats.Western blot was used to detect the effects of the serum contai-ning drug on the expression of endoplasmic reticulum stress-related proteins(PERK,eIF-2α,ATF-4,GRP78,CHOP)and pyroptosis related proteins(NL-RP1,caspase-1,cleaved caspase-1,GSDMD,GSDMD-N).MTT assay was used to detect the activity of Hep-aRG cells after treatment with the liver homogenate containing ESMAE from SD rats.Results Twenty percent serum containing drug significantly decreased the viability of HepaRG cells,with the cells exhibiting swelling,rupture,and vesicle-like pyroptosis.The ex-pression levels of endoplasmic reticulum stress-related proteins PERK,eIF-2α,ATF-4,GRP78,and CHOP significantly increased.The expression levels of pro-teins involved in the NLRP1-mediated classical pyrop-tosis pathway were significantly increased.Finally the liver homogenate containing drug decreased the cell ac-tivity,and cells exhibited swelling,rupture,and vesicle-like pyroptosis.Conclusions After administration of ESMAE,the serum containing drug and the liver ho-mogenate containing drug of rats show toxicity to Hep-aRG cells,and can induce endoplasmic reticulum stress and activate the NLRP1/caspase-1/GSDMD pyroptosis pathway in HepaRG cells.

Objective:To investigate the differences in clinical outcomes of septic children with varying serum zinc levels,and to analyze the relationship between reduced serum zinc levels and organ dysfunction as well as 28-day mortality in septic children.Methods:This study conducted a retrospective analysis of clinical data from pediatric patients diagnosed with sepsis or septic shock in the Department of critical care medicine of the children's Hospital of Soochow University between January 2017 and December 2022.Clinical characteristics,organ dysfunction,and prognosis were compared between two groups:children with low serum zinc levels and those with normal zinc levels.Results:The serum zinc level of septic children within 24 hours of admission was 9.60(5.52,13.80)μmol/L,with 50.54%(94/186)of the children exhibiting low serum zinc levels(<10.07 μmol/L).Compared to the normal serum zinc group,the low serum zinc group had a significantly lower Pediatric Critical Illness Score(PCIS)[(78.71±9.35)vs.(85.12±8.51),P=0.005]and higher 28-day mortality(46.80%vs.14.13%,P<0.001).The low serum zinc group also had a higher proportion of invasive mechanical ventilation(64.89%vs.47.82%,P=0.019),renal replacement therapy(15.59%vs.3.26%,P=0.003),and use of vasoactive drugs(56.38%vs.30.43%,P<0.001).The rate of underlying conditions in the low serum zinc group was significantly higher than that in the normal serum zinc group(57.44%vs.36.95%,P=0.005).Additionally,the low serum zinc group had a higher incidence of disseminated intravascular coagulation(DIC),respiratory failure,acute kidney injury,shock,and multiple organ dysfunction syndrome(MODS)compared to the normal serum zinc group(P<0.05).Serum zinc levels had predictive value for 28-day mortality in septic children(AUC=0.813;95%CI:0.725～0.902;P<0.001).A serum zinc level of less than 6.950 μmol/L predicted the death of septic children with a sensitivity of 0.618 and a specificity of 0.902.Conclusion:Sepsis in children is commonly associated with low serum zinc levels,especially in those with underlying conditions such as hematologic and oncologic disorders.Sepsis patients hypozincemia with a higher incidence of DIC,respiratory failure,acute kidney injury,shock,and MODS.A serum zinc level below 6.95 μmol/L serves as a significant predictor of 28-day mortality in children with severe sepsis.