Objective To investigate the ameliorative effects of 4-hydroxybenzyl alcohol(4HBA)on energy metabolism in ischemic astrocytes(ASC).Methods Primary ASC were isolated from rat cortical tissue,and then randomly divided into control,oxygen-glucose deprivation/reoxygenation(OGD/R)group,low-,medium-and high-dose 4HBA groups(50,100 and 200 μmol/L),and edaravone group.Except for the control group,OGD/R injury models were established in a tri-gas incubator.The following parameters were measured:cell mortality,adenosine triphosphate(ATP)content,activities of mitochondrial ATP enzymes(Na+/K+-ATPase and Ca2+/Mg2+-ATPase),activities of mitochondrial complexes Ⅰ-Ⅴ,and expression of energy metabolism-related genes.Results Compared with the control group,the OGD/R group showed significantly higher cell mortality[(9.49±0.85)％vs(0.00±0.00)％,P＜0.01],along with obviously decreased ATP content,Na+/K+-ATPase activity,Ca2+/Mg2+-ATPase activity,and mitochondrial complex Ⅰ-V activities(P＜0.01).Compared with the OGD/R group,three 4HBA groups as well as the edaravone group exhibited significantly reduced cell mortality and elevated ATP content,Na+/K+-ATPase activity,and complex Ⅰ,Ⅲ,and V activities.The high-and medium-dose 4HBA groups and the edaravone group demonstrated increased Ca2+/Mg2+-ATPase activity,and the high-dose 4HBA group and edaravone group exhibited more enhanced complexⅡ activity,and the three 4HBA groups had elevated complex Ⅳ activity(P＜0.05,P＜0.01).Conclusion 4HBA exerts protective effect against ischemic injury in ASC by mitigating energy metabolism dysfunction and reducing cell death.

Objective To investigate the neuroprotective effects and mechanisms of the alcoholic extract of the Yi drug(Rubia yunnanensis)on the Alzheimer's disease(AD)model of Caenorhabditis elegans(C.elegans).Methods Based on a transgenic C.elegans model of AD,the efficacy and mechanism of alcoholic extracts of Rubia yunnanensis against β-amyloid(Aβ)-induced toxic effects were explored.C.elegans were synchronized and divided into a control group and low and high dose groups of the alcoholic extract of Rubia yunnanensis to study the effects of the alcoholic extract of Rubia yunnanensis on C.elegans paralysis,lifespan,lipofuscin levels,behavior,growth and development,heat stress and oxidative stress,ROS levels,and Aβ aggregation,as well as the key transcription factors in the insulin/insulin-like growth factor 1 signaling pathway were investigated by GFP reporter gene worm assay and RNAi assay DAF-16 expression.Results Compared with the Control group,the intervention of the alcoholic extracts of Rubia yunnanensis could effectively improve the paralyzed phenotype of C.elegans(P<0.001),prolong the lifespan of C.elegans and reduce the level of lipofuscin in C.elegans significantly(P<0.001),improve the mobility of C.elegans significantly(P<0.001),and improve its resistance to oxidative stress and heat stress damage significantly(P<0.001),and reduce the level of ROS in C.elegans significantly(P<0.001),and reduce the deposition of Aβ protein in the head of C.elegans significantly(P<0.01),and had no effect on the growth and development of C.elegans(P>0.05),and also promoted the nuclear translocation of DAF-16 in the reporter gene C.elegans TJ356,and the effect of the alcoholic extracts of Rubia yunnanensis in delaying the degree of paralysis of C.elegans was lost after silencing the expression of C.elegans DAF-16 by RNAi,whereas the L4440 empty vector control group in the administration of Rubia yunnanensis alcohol extract significantly reduced the level of C.elegans paralysis(P<0.001).Conclusion The alcoholic extract of Rubia yunnanensis can significantly ameliorate the paralyzed phenotype in the C.elegans model of Alzheimer's disease with certain antioxidant,anti-aging and anti-Aβ protein activities,and the mechanism may be related to the activation of DAF-16,a downstream transcription factor of the insulin signaling pathway.

[Objective]To explore the academic characteristics and ideological viewpoints in Nanjing Zhengyi,in order to provide a novel perspective and inspiration for the study of traditional Chinese medicine(TCM)theoretical classics.[Methods]Using Nanjing Zhengyi as the research subject,a literature review method was employed to systematically analyze YE Lin's academic viewpoints.By comparing annotated versions of Nanjing and integrating theories from both Chinese and western medicine,medical and I Ching thoughts,and theory of five movements and six climates,a multidimensional analysis was conducted on his academic perspectives and exegetical features.[Results]YE Lin employed western anatomical knowledge to elucidate TCM concepts such as Ying-Wei-Qi-blood,emphasizing TCM as the foundation while dialectically incorporating western medical knowledge.He clarified the anatomical localization of the pericardium,tri-Jiao and Mingmen,integrating Qi transformation theory to form an innovative model.Utilizing I Ching,he interpreted core mechanisms such as"fire entering water to transform Qi"and"Mingmen was the Taichi of the body".Additionally,he applied the five phases theory and the five movements and six climates theory to explore the principles of visceral Yin-Yang and epidemic diseases.[Conclusion]Nanjing Zhengyi represents an exemplary annotated edition of Nanjing with integration of Chinese and western medicine,medical and I Ching philosophy,and the five movements and six climates theory,showcasing YE Lin's academic approach which is grounded in TCM while incorporating western learning and integrating medical theories with I Ching principles.His theoretical innovations provide new perspectives for TCM classical research,warranting further study and clinical validation by future scholars.

Objective:To investigate the pattern of Mesothelin(MSLN)-specific T-cell(CTLs)immune reconstitution after allogeneic hematopoietic stem cell transplantation.Methods:Two cases of Mesothelin-positive patients with acute myeloid leukemia(AML)were screened,and the number of MSLN-CTLs and its subpopulations,the expression of surface PD-1/CTLA-4/TIM-3 im-mune exhaustion molecules,and the functions of secreted cytokines TNF-α and IFN-γ were monitored by flow cytometry after trans-plantation.They were compared with WT1-CTLs and cytomegalovirus(CMV)CTLs.Meanwhile,the relationship between CTL recon-stitution and Minimal residual disease(MRD)and leukemia recurrence was analyzed.Results:After transplantation,MSLN-CTLs,WT1-CTLs and CMV-CTLs can be detected,and intracellular factors were secreted.The phenotypes of WT1 specific CD8+T cells,MSLN specific CD8+T cells and CMV specific CD8+T cells were mainly TEM subsets and TEMRA subsets,and the TEM subsets of CMV specific CD8+T cells were more obvious.Compared with CMV-CTLs,the proportions of T Naive,TCM and TEMRA subsets were relatively higher in MSLN-specific CD8+T cells and WT1-specific CD8+T cells,and the expression levels of PD-1,CTLA-4 and TIM-3 were higher in MSLN-CTLs and WT1-CTLs.The dynamic changes of MSLN-CTLs and WT1-CTLs after transplantation were related to leukemia load and the chimerism rate of donor and recipient.Conclusion:After allogeneic hematopoietic stem cell transplantation,im-mune recovery to MSLN is found,which is different from WT1-CTLs and CMV-CTLs.

Objective:This study aimed to evaluate whether intra-articular delivery of conditioned medium(CM)derived from stem cells of human exfoliated deciduous teeth(SHED)could influence the progression of condylar cartilage degeneration in a rat model of temporomandibular joint osteoarthritis(TMJ OA).Methods:Sixty 8-week-old male SD rats were randomly divided into control group(CON group),intraarticular injection of MIA induced TMJ OA model group(MIA group),and injection of SHED condi-tioned medium 1 week after MIA modeling for treatment group(SHED-CM group),with 20 animals in each group.Histological sec-tions,HE,Safranine O-solid green staining,Col Ⅱ immunohistochemical staining,and TUNEL staining were performed 2 and 4 weeks after the start of treatment.Western blotting and RT-qPCR were used to detect the key molecules of apoptosis,cleaved-CASP3,BAX and BCL2,pro-inflammatory related factors IL-1β,IL-6,TNFα,MMP3,ADAMTS5,and the MAPK pathway-related molecules p-ERK,ERK,p-P38 and P38.Results:Compared with the CON group and SHED-CM group,the condyle chondrocytes in the MIA group had disordered arrange-ment,interrupted layers,significantly thickened fibrous layers(P＜0.001),and significantly increased Mankin's OA histological score(P＜0.001).In the MIA group,both the Safranin O-positive area ratio and the proportion of ColⅡ-positive regions were markedly reduced compared with the CON and SHED-CM groups(P＜0.001).Conversely,the proportion of TUNEL-positive cells was substantially higher than in the other two groups(both P＜0.001).Western blot analysis further demonstrated that apoptotic markers(cleaved-CASP3,BAX/BCL2)and MAPK pathway-related proteins(p-ERK,ERK,p-P38,P38)were expressed at significantly elevated levels in the MIA group relative to CON and SHED-CM groups(BAX/BCL2:P＜0.05;cleaved-CASP3:P＜0.01;p-P38/P38:P＜0.001;p-ERK/ERK:P＜0.01).Similarly,qRT-PCR revealed upregulated expression of inflammatory mediators,including IL-1 β(P＜0.001),IL-6(P＜0.01),TNFα(P＜0.01),MMP3(P＜0.001),and ADAMTS5(P＜0.05),in the MIA group compared with the CON and SHED-CM groups.Conclusion:SHED-CM treatment can ef-fectively reverse MIA-induced condylar cartilage degeneration of TMJ OA in rats.

Objective:This study aimed to evaluate whether intra-articular delivery of conditioned medium(CM)derived from stem cells of human exfoliated deciduous teeth(SHED)could influence the progression of condylar cartilage degeneration in a rat model of temporomandibular joint osteoarthritis(TMJ OA).Methods:Sixty 8-week-old male SD rats were randomly divided into control group(CON group),intraarticular injection of MIA induced TMJ OA model group(MIA group),and injection of SHED condi-tioned medium 1 week after MIA modeling for treatment group(SHED-CM group),with 20 animals in each group.Histological sec-tions,HE,Safranine O-solid green staining,Col Ⅱ immunohistochemical staining,and TUNEL staining were performed 2 and 4 weeks after the start of treatment.Western blotting and RT-qPCR were used to detect the key molecules of apoptosis,cleaved-CASP3,BAX and BCL2,pro-inflammatory related factors IL-1β,IL-6,TNFα,MMP3,ADAMTS5,and the MAPK pathway-related molecules p-ERK,ERK,p-P38 and P38.Results:Compared with the CON group and SHED-CM group,the condyle chondrocytes in the MIA group had disordered arrange-ment,interrupted layers,significantly thickened fibrous layers(P＜0.001),and significantly increased Mankin's OA histological score(P＜0.001).In the MIA group,both the Safranin O-positive area ratio and the proportion of ColⅡ-positive regions were markedly reduced compared with the CON and SHED-CM groups(P＜0.001).Conversely,the proportion of TUNEL-positive cells was substantially higher than in the other two groups(both P＜0.001).Western blot analysis further demonstrated that apoptotic markers(cleaved-CASP3,BAX/BCL2)and MAPK pathway-related proteins(p-ERK,ERK,p-P38,P38)were expressed at significantly elevated levels in the MIA group relative to CON and SHED-CM groups(BAX/BCL2:P＜0.05;cleaved-CASP3:P＜0.01;p-P38/P38:P＜0.001;p-ERK/ERK:P＜0.01).Similarly,qRT-PCR revealed upregulated expression of inflammatory mediators,including IL-1 β(P＜0.001),IL-6(P＜0.01),TNFα(P＜0.01),MMP3(P＜0.001),and ADAMTS5(P＜0.05),in the MIA group compared with the CON and SHED-CM groups.Conclusion:SHED-CM treatment can ef-fectively reverse MIA-induced condylar cartilage degeneration of TMJ OA in rats.

Objective To investigate the neuroprotective effects and mechanisms of the alcoholic extract of the Yi drug(Rubia yunnanensis)on the Alzheimer's disease(AD)model of Caenorhabditis elegans(C.elegans).Methods Based on a transgenic C.elegans model of AD,the efficacy and mechanism of alcoholic extracts of Rubia yunnanensis against β-amyloid(Aβ)-induced toxic effects were explored.C.elegans were synchronized and divided into a control group and low and high dose groups of the alcoholic extract of Rubia yunnanensis to study the effects of the alcoholic extract of Rubia yunnanensis on C.elegans paralysis,lifespan,lipofuscin levels,behavior,growth and development,heat stress and oxidative stress,ROS levels,and Aβ aggregation,as well as the key transcription factors in the insulin/insulin-like growth factor 1 signaling pathway were investigated by GFP reporter gene worm assay and RNAi assay DAF-16 expression.Results Compared with the Control group,the intervention of the alcoholic extracts of Rubia yunnanensis could effectively improve the paralyzed phenotype of C.elegans(P<0.001),prolong the lifespan of C.elegans and reduce the level of lipofuscin in C.elegans significantly(P<0.001),improve the mobility of C.elegans significantly(P<0.001),and improve its resistance to oxidative stress and heat stress damage significantly(P<0.001),and reduce the level of ROS in C.elegans significantly(P<0.001),and reduce the deposition of Aβ protein in the head of C.elegans significantly(P<0.01),and had no effect on the growth and development of C.elegans(P>0.05),and also promoted the nuclear translocation of DAF-16 in the reporter gene C.elegans TJ356,and the effect of the alcoholic extracts of Rubia yunnanensis in delaying the degree of paralysis of C.elegans was lost after silencing the expression of C.elegans DAF-16 by RNAi,whereas the L4440 empty vector control group in the administration of Rubia yunnanensis alcohol extract significantly reduced the level of C.elegans paralysis(P<0.001).Conclusion The alcoholic extract of Rubia yunnanensis can significantly ameliorate the paralyzed phenotype in the C.elegans model of Alzheimer's disease with certain antioxidant,anti-aging and anti-Aβ protein activities,and the mechanism may be related to the activation of DAF-16,a downstream transcription factor of the insulin signaling pathway.

[Objective]To explore the academic characteristics and ideological viewpoints in Nanjing Zhengyi,in order to provide a novel perspective and inspiration for the study of traditional Chinese medicine(TCM)theoretical classics.[Methods]Using Nanjing Zhengyi as the research subject,a literature review method was employed to systematically analyze YE Lin's academic viewpoints.By comparing annotated versions of Nanjing and integrating theories from both Chinese and western medicine,medical and I Ching thoughts,and theory of five movements and six climates,a multidimensional analysis was conducted on his academic perspectives and exegetical features.[Results]YE Lin employed western anatomical knowledge to elucidate TCM concepts such as Ying-Wei-Qi-blood,emphasizing TCM as the foundation while dialectically incorporating western medical knowledge.He clarified the anatomical localization of the pericardium,tri-Jiao and Mingmen,integrating Qi transformation theory to form an innovative model.Utilizing I Ching,he interpreted core mechanisms such as"fire entering water to transform Qi"and"Mingmen was the Taichi of the body".Additionally,he applied the five phases theory and the five movements and six climates theory to explore the principles of visceral Yin-Yang and epidemic diseases.[Conclusion]Nanjing Zhengyi represents an exemplary annotated edition of Nanjing with integration of Chinese and western medicine,medical and I Ching philosophy,and the five movements and six climates theory,showcasing YE Lin's academic approach which is grounded in TCM while incorporating western learning and integrating medical theories with I Ching principles.His theoretical innovations provide new perspectives for TCM classical research,warranting further study and clinical validation by future scholars.

Objective:To investigate the pattern of Mesothelin(MSLN)-specific T-cell(CTLs)immune reconstitution after allogeneic hematopoietic stem cell transplantation.Methods:Two cases of Mesothelin-positive patients with acute myeloid leukemia(AML)were screened,and the number of MSLN-CTLs and its subpopulations,the expression of surface PD-1/CTLA-4/TIM-3 im-mune exhaustion molecules,and the functions of secreted cytokines TNF-α and IFN-γ were monitored by flow cytometry after trans-plantation.They were compared with WT1-CTLs and cytomegalovirus(CMV)CTLs.Meanwhile,the relationship between CTL recon-stitution and Minimal residual disease(MRD)and leukemia recurrence was analyzed.Results:After transplantation,MSLN-CTLs,WT1-CTLs and CMV-CTLs can be detected,and intracellular factors were secreted.The phenotypes of WT1 specific CD8+T cells,MSLN specific CD8+T cells and CMV specific CD8+T cells were mainly TEM subsets and TEMRA subsets,and the TEM subsets of CMV specific CD8+T cells were more obvious.Compared with CMV-CTLs,the proportions of T Naive,TCM and TEMRA subsets were relatively higher in MSLN-specific CD8+T cells and WT1-specific CD8+T cells,and the expression levels of PD-1,CTLA-4 and TIM-3 were higher in MSLN-CTLs and WT1-CTLs.The dynamic changes of MSLN-CTLs and WT1-CTLs after transplantation were related to leukemia load and the chimerism rate of donor and recipient.Conclusion:After allogeneic hematopoietic stem cell transplantation,im-mune recovery to MSLN is found,which is different from WT1-CTLs and CMV-CTLs.

Objective To investigate the ameliorative effects of 4-hydroxybenzyl alcohol(4HBA)on energy metabolism in ischemic astrocytes(ASC).Methods Primary ASC were isolated from rat cortical tissue,and then randomly divided into control,oxygen-glucose deprivation/reoxygenation(OGD/R)group,low-,medium-and high-dose 4HBA groups(50,100 and 200 μmol/L),and edaravone group.Except for the control group,OGD/R injury models were established in a tri-gas incubator.The following parameters were measured:cell mortality,adenosine triphosphate(ATP)content,activities of mitochondrial ATP enzymes(Na+/K+-ATPase and Ca2+/Mg2+-ATPase),activities of mitochondrial complexes Ⅰ-Ⅴ,and expression of energy metabolism-related genes.Results Compared with the control group,the OGD/R group showed significantly higher cell mortality[(9.49±0.85)％vs(0.00±0.00)％,P＜0.01],along with obviously decreased ATP content,Na+/K+-ATPase activity,Ca2+/Mg2+-ATPase activity,and mitochondrial complex Ⅰ-V activities(P＜0.01).Compared with the OGD/R group,three 4HBA groups as well as the edaravone group exhibited significantly reduced cell mortality and elevated ATP content,Na+/K+-ATPase activity,and complex Ⅰ,Ⅲ,and V activities.The high-and medium-dose 4HBA groups and the edaravone group demonstrated increased Ca2+/Mg2+-ATPase activity,and the high-dose 4HBA group and edaravone group exhibited more enhanced complexⅡ activity,and the three 4HBA groups had elevated complex Ⅳ activity(P＜0.05,P＜0.01).Conclusion 4HBA exerts protective effect against ischemic injury in ASC by mitigating energy metabolism dysfunction and reducing cell death.