With the rapid development of artificial intelligence, computational pathology has been seamlessly integrated into the entire clinical workflow, which encompasses diagnosis, treatment, prognosis, and biomarker discovery. This integration has significantly enhanced clinical accuracy and efficiency while reducing the workload for clinicians. Traditionally, research in this field has depended on the collection and labeling of large datasets for specific tasks, followed by the development of task-specific computational pathology models. However, this approach is labor intensive and does not scale efficiently for open-set identification or rare diseases. Given the diversity of clinical tasks, training individual models from scratch to address the whole spectrum of clinical tasks in the pathology workflow is impractical, which highlights the urgent need to transition from task-specific models to foundation models (FMs). In recent years, pathological FMs have proliferated. These FMs can be classified into three categories, namely, pathology image FMs, pathology image-text FMs, and pathology image-gene FMs, each of which results in distinct functionalities and application scenarios. This review provides an overview of the latest research advancements in pathological FMs, with a particular emphasis on their applications in oncology. The key challenges and opportunities presented by pathological FMs in precision oncology are also explored.
Humans ; Precision Medicine/methods* ; Medical Oncology/methods* ; Artificial Intelligence ; Neoplasms/pathology* ; Computational Biology/methods*

Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards* ; Humans ; Consensus ; Drugs, Chinese Herbal/therapeutic use* ; Surveys and Questionnaires

Beta-amylases (BAMs), key enzymes in starch hydrolysis, play an important role in plant growth, development, and resistance to abiotic stress. To mine the saline-alkali tolerance-related BAM genes in alfalfa (Medicago sativa L.), we identified MsBAM genes in the whole genome. The physicochemical properties, phylogeny, gene structures, conserved motifs, secondary structures, promoter cis-acting elements, chromosome localization, and gene replication relationships of BAM gene family members were analyzed. RNA-seq and quantitative real-time PCR (qRT-PCR) were employed to analyze the expression patterns of BAM family members under saline-alkali stress. The results showed that 54 BAM genes were identified in the genome, which were classified into 8 subgroups according to the phylogenetic tree. The members of the same subgroup had similar gene structures except that those of subgroups 1 and 7 had large differences. Conserved motif analysis showed that all MsBAM proteins had a typical glycohydrolysis domain. The chromosome localization analysis showed that MsBAM gene family members were unevenly distributed on 27 chromosomes. The duplication of gene segments led to the increase in BAM gene number in alfalfa. The promoters of BAM genes contained a large number of elements in response to plant hormones and stress. Transcriptome data and qRT-PCR results showed that the expression levels of most MsBAM genes were up-regulated in response to saline-alkali stress. Under the saline-alkali stress, the expression levels of 28 genes, including MsBAM6, were up-regulated on days 1 and 7, and those of 5 genes, including MsBAM9, were up-regulated by over 2 folds. In addition, under salt-alkali stress, BAM activity and soluble sugar content were significantly increased. These results indicate that BAM genes play a key role in alfalfa in response to saline-alkali stress, laying a foundation for further research in this field.
Medicago sativa/physiology* ; beta-Amylase/metabolism* ; Phylogeny ; Gene Expression Regulation, Plant ; Stress, Physiological/genetics* ; Multigene Family ; Alkalies ; Plant Proteins/genetics*

Objective:To generate a chronic lymphocytic leukemia (CLL) mouse model with intact immune competence and short latency by adoptively transferring (AT) splenocytes from immunoglobulin heavy-chain enhancer-driven T-cell leukemia/lymphoma 1 (Eμ-TCL1) transgenic donors into wild-type (WT) recipients.Methods:Specific pathogen-free C57BL/6J WT mice and H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice were utilized. The H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice were generated using CRISPR/Cas9 technology, and their genotypes were confirmed by PCR. Experimental animals were randomly divided into an adoptive transfer (AT) group and a WT control group ( n=10 per group). Mice in the AT group received an intraperitoneal injection of splenocytes from H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice. The weight and general condition of the mice were monitored. Mice were euthanized by cervical dislocation at 9 weeks post-transplantation. The CLL model was validated using key indicators, including pathological manifestations, changes in peripheral blood leukocyte counts, and immunophenotype. Results:AT group mice exhibited significantly increased spleen weight [ (0.92±0.16) g vs (0.06±0.01) g in WT group, P<0.05] and liver weight [ (2.11±0.56) g vs (1.42±0.13) g in WT group, P=0.006], indicative of marked splenomegaly and hepatomegaly. The peripheral blood leukocyte count was significantly higher in the AT group [ (124.33±8.74) ×10 9/L] compared to the WT group [ (5.55±1.67) ×10 9/L] ( P=0.002). Similarly, the percentage of peripheral blood B lymphocytes was markedly increased in the AT group versus the WT group [ (69.13±6.88) % vs (39.78±5.94) %, P<0.05]. Histopathological examination revealed CLL manifestations in the spleen, lymph nodes, and bone marrow of AT group mice, with significant lymphocytic infiltration observed in the liver, lung, and kidney tissues. Flow cytometry analysis showed that the percentages of CD19 +CD5 + B lymphocytes among total lymphocytes in peripheral blood, bone marrow, and spleen of the AT group were (61.37±9.92) %, (28.61± 7.08) %, and (86.03±5.78) %, respectively. These were significantly higher (all P<0.05) than in the WT group [ (4.51±1.32) %, (5.58±1.46) %, and (14.33±3.20) %]. Furthermore, these CLL-like cells in the AT group were positive for CD43 and CD200, but showed lower expression of CD20, CD22, and CD79b compared to WT B cells. Conclusion:Adoptive transfer of splenocytes from Eμ-TCL1 transgenic mice successfully established a CLL mouse model with a relatively short latency period. This model represents a valuable preclinical tool for investigating CLL and related pathologies.

Objective:To investigate effect of rapamycin on proliferation of human chronic lymphoblastic leukemia tumor cell MEC-1 in vitro.Methods:Human γδT cells were cultured and expanded in vitro.Human γδT cells percentage was detected by flow cytometry on the 10th day.γδT cells were treated with different concentrations of rapamycin(0,50,100 and 200 nmol/L).After 48 h of intervention,IL-17,IL-12,IFN-γ and TNF-α contents secreted by γδT cells were determined by ELISA,and AKT/mTOR protein expression was detected by Western blot.Supernatant of γδT was collected and co-cultured with MEC-1 indirectly.Co-cultured experi-mental was divided into DMSO group,rapamycin group,γδT group,rapamycin treated γδT group and control group.Survival rate of MEC-1 cells in each group was determined by CCK-8 after 48 h of culture.Results:IL-12,IFN-γ and TNF-α levels secreted by γδT cells were the highest when rapamycin was 100 nmol/L,which was significantly higher than other groups(P<0.05).When rapamycin was 50 nmol/L,IL-17 secretion level of γδT cells was the highest,and IL-17 level secreted by γδT cells gradually decreased with increase of rapamycin concentration.Survival rates of MEC-1 cells in supernatant of DMSO group,rapamycin group,rapamycin treated γδT cell group and γδT cell group were(93.48±2.52)%,(71.44±4.31)%,(83.93±1.54)%and(57.97±2.47)%,whose differences were statistically significant compared with control group(P<0.05).Compared with control group,mTOR protein level of γδT cells was decreased in all rapamycin treated groups.Protein expressions of upstream AKT,pAKT and downstream specific transcription factor 4EBP-1 were decreased with increase of rapamycin concentration(P<0.05).Conclusion:There is a dose-effect relationship between rapamycin concentration and IL-17,IL-12,IFN-γ,TNF-α secretions by γδT cells.Rapamycin and γδT cells can inhibit pro-liferation of MEC-1 cells,and 100 nmol/L rapamycin can enhance inhibitory effect of γδT cells on proliferation of MEC-1 cells.Rapa-mycin can weaken AKT pathway on γδT cells,and rapamycin combined with γδT cells plays a positive role in anti-tumor application of leukemia.

Objective:To explore the difference of type 2 innate lymphoid cell(ILC2),IL-9 and related cytokines between chronic lymphocytic leukemia(CLL)patients and normal individuals,as well as the correlation between ILC2 and IL-9 and other cytokines in CLL patients.Methods:Flow cytometry was used to detect the expression of ILC2 and regulatory T cells(Tregs)in peripheral blood of 26 CLL patients at initial diagnosis and 10 healthy controls.RT-qPCR was used to detected IFN-γ,TGF-β,IL-9 and IL-10 mRNA in peripheral blood mononuclear cell(PBMC).ELISA was used to detect serum IFN-γ,TNF-α,TGF-β,IL-9,IL-10 and IL-21.ILC2 and IL-9 were observed in the cervical lymph node of 12 CLL patients at initial diagnosis and 12 patients with reactive lymphoid hyperplasias by multiplex indirect immunofluorescence staining.Spearman test was used to analyze the correlation between peripheral blood ILC2 and IL-9,IL-9 and IL-21,IFN-γ mRNA and IL-10 mRNA in CLL patients.Pearson test was used to analyze the correlation between TNF-α and TGF-β in CLL patients.Results:Compared with control group,the proportions of ILC2 and Tregs were significantly increased in CLL group(both P＜0.05).The mRNA expressions of IFN-γ,IL-9,IL-10 and TGF-β in PBMCs of CLL patients were significantly increased(all P＜0.05).In CLL patients,the expressions of IFN-γ,TNF-α,TGF-β,IL-9 and IL-10 in serum were significantly increased(all P＜0.01),while IL-21 slightly increased without statistical difference(P＞0.05).In CLL patients,the peripheral blood ILC2 was positively related to IL-9(r=0.56),IL-9 was positively related to IL-21(r=0.397),IFN-γ mRNA was positively related to IL-10 mRNA(r=0.623),and TNF-α was positively related to TGF-β(r=0.577).Compared with reactive lymphoid hyperplasias patients,the mean fluorescence intensities of GAT A3 and CRTH2 representing ILC2 and IL-9 in cervical lymph nodes were significantly increased in the CLL group(all P＜0.001),and showed colocalization.Conclusion:In CLL patients,the proportions of ILC2 and IL-9 in peripheral blood and lymph nodes increase,and ILC2 and IL-9 show colocalization in lymph nodes.There is a positive correlation between ILC2 and IL-9 in the peripheral blood of CLL patients,the ability of ILC2 to secrete IL-9 is increased,and ILC2 may affect the occurrence and development of CLL through IL-9.

ObjectiveTo investigate the anti-inflammatory, antioxidant, and goblet cell-stimulating effects of a suspension of Ophiopogon japonicus (L. f.) Ker Gawl. (O. japonicus, Mai Dong) extract combined with hyaluronic acid (HA) in the mouse model with dry eye disease (DED).MethodsA DED mouse model was induced using benzalkonium chloride (BAK), followed by treatment with O. japonicus extract-containing eye drops at varying concentrations. Experimental groups included a normal control, a DED model control, a positive control, and an O. japonicus extract-treated group. Corneal fluorescein staining and tear break-up time (TBUT) were used to assess tear film stability and ocular surface integrity. Enzyme-linked immunosorbent assay (ELISA) measured inflammatory factor levels in corneal and conjunctival tissues, whereas Western blot (WB) analyzed key antioxidant and inflammatory markers, including nuclear factor erythroid 2-related factor (2Nrf2) and heme oxygenase 1 (HO-1). Periodic acid-schiff (PAS) staining and immunofluorescence were used to evaluate goblet cell density and mucin secretion.ResultsO. japonicus extract significantly improved corneal damage, reduced fluorescein staining scores, prolonged TBUT, and increased tear secretion. It downregulated inflammatory markers, including interleukin-8 (IL-8), interleukin-1β (IL-1β), and interferon-γ (IFN-γ) while upregulating Nrf2, HO-1, and the interleukin-13 (IL-13)/IFN-γ ratio, alleviating oxidative stress and inflammation. PAS staining showed increased conjunctival goblet cell density and restored mucin secretion, enhancing tear film stability.ConclusionO. japonicus extract demonstrated significant anti-inflammatory, antioxidant, and goblet cell-stimulating effects in a DED model, with good biocompatibility and promising therapeutic potential. Future research should optimize extraction processes and validate their efficacy and safety in clinical settings.

Objective:To explore the difference of type 2 innate lymphoid cell(ILC2),IL-9 and related cytokines between chronic lymphocytic leukemia(CLL)patients and normal individuals,as well as the correlation between ILC2 and IL-9 and other cytokines in CLL patients.Methods:Flow cytometry was used to detect the expression of ILC2 and regulatory T cells(Tregs)in peripheral blood of 26 CLL patients at initial diagnosis and 10 healthy controls.RT-qPCR was used to detected IFN-γ,TGF-β,IL-9 and IL-10 mRNA in peripheral blood mononuclear cell(PBMC).ELISA was used to detect serum IFN-γ,TNF-α,TGF-β,IL-9,IL-10 and IL-21.ILC2 and IL-9 were observed in the cervical lymph node of 12 CLL patients at initial diagnosis and 12 patients with reactive lymphoid hyperplasias by multiplex indirect immunofluorescence staining.Spearman test was used to analyze the correlation between peripheral blood ILC2 and IL-9,IL-9 and IL-21,IFN-γ mRNA and IL-10 mRNA in CLL patients.Pearson test was used to analyze the correlation between TNF-α and TGF-β in CLL patients.Results:Compared with control group,the proportions of ILC2 and Tregs were significantly increased in CLL group(both P＜0.05).The mRNA expressions of IFN-γ,IL-9,IL-10 and TGF-β in PBMCs of CLL patients were significantly increased(all P＜0.05).In CLL patients,the expressions of IFN-γ,TNF-α,TGF-β,IL-9 and IL-10 in serum were significantly increased(all P＜0.01),while IL-21 slightly increased without statistical difference(P＞0.05).In CLL patients,the peripheral blood ILC2 was positively related to IL-9(r=0.56),IL-9 was positively related to IL-21(r=0.397),IFN-γ mRNA was positively related to IL-10 mRNA(r=0.623),and TNF-α was positively related to TGF-β(r=0.577).Compared with reactive lymphoid hyperplasias patients,the mean fluorescence intensities of GAT A3 and CRTH2 representing ILC2 and IL-9 in cervical lymph nodes were significantly increased in the CLL group(all P＜0.001),and showed colocalization.Conclusion:In CLL patients,the proportions of ILC2 and IL-9 in peripheral blood and lymph nodes increase,and ILC2 and IL-9 show colocalization in lymph nodes.There is a positive correlation between ILC2 and IL-9 in the peripheral blood of CLL patients,the ability of ILC2 to secrete IL-9 is increased,and ILC2 may affect the occurrence and development of CLL through IL-9.

Objective:To generate a chronic lymphocytic leukemia (CLL) mouse model with intact immune competence and short latency by adoptively transferring (AT) splenocytes from immunoglobulin heavy-chain enhancer-driven T-cell leukemia/lymphoma 1 (Eμ-TCL1) transgenic donors into wild-type (WT) recipients.Methods:Specific pathogen-free C57BL/6J WT mice and H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice were utilized. The H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice were generated using CRISPR/Cas9 technology, and their genotypes were confirmed by PCR. Experimental animals were randomly divided into an adoptive transfer (AT) group and a WT control group ( n=10 per group). Mice in the AT group received an intraperitoneal injection of splenocytes from H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice. The weight and general condition of the mice were monitored. Mice were euthanized by cervical dislocation at 9 weeks post-transplantation. The CLL model was validated using key indicators, including pathological manifestations, changes in peripheral blood leukocyte counts, and immunophenotype. Results:AT group mice exhibited significantly increased spleen weight [ (0.92±0.16) g vs (0.06±0.01) g in WT group, P<0.05] and liver weight [ (2.11±0.56) g vs (1.42±0.13) g in WT group, P=0.006], indicative of marked splenomegaly and hepatomegaly. The peripheral blood leukocyte count was significantly higher in the AT group [ (124.33±8.74) ×10 9/L] compared to the WT group [ (5.55±1.67) ×10 9/L] ( P=0.002). Similarly, the percentage of peripheral blood B lymphocytes was markedly increased in the AT group versus the WT group [ (69.13±6.88) % vs (39.78±5.94) %, P<0.05]. Histopathological examination revealed CLL manifestations in the spleen, lymph nodes, and bone marrow of AT group mice, with significant lymphocytic infiltration observed in the liver, lung, and kidney tissues. Flow cytometry analysis showed that the percentages of CD19 +CD5 + B lymphocytes among total lymphocytes in peripheral blood, bone marrow, and spleen of the AT group were (61.37±9.92) %, (28.61± 7.08) %, and (86.03±5.78) %, respectively. These were significantly higher (all P<0.05) than in the WT group [ (4.51±1.32) %, (5.58±1.46) %, and (14.33±3.20) %]. Furthermore, these CLL-like cells in the AT group were positive for CD43 and CD200, but showed lower expression of CD20, CD22, and CD79b compared to WT B cells. Conclusion:Adoptive transfer of splenocytes from Eμ-TCL1 transgenic mice successfully established a CLL mouse model with a relatively short latency period. This model represents a valuable preclinical tool for investigating CLL and related pathologies.

Objective:To investigate effect of rapamycin on proliferation of human chronic lymphoblastic leukemia tumor cell MEC-1 in vitro.Methods:Human γδT cells were cultured and expanded in vitro.Human γδT cells percentage was detected by flow cytometry on the 10th day.γδT cells were treated with different concentrations of rapamycin(0,50,100 and 200 nmol/L).After 48 h of intervention,IL-17,IL-12,IFN-γ and TNF-α contents secreted by γδT cells were determined by ELISA,and AKT/mTOR protein expression was detected by Western blot.Supernatant of γδT was collected and co-cultured with MEC-1 indirectly.Co-cultured experi-mental was divided into DMSO group,rapamycin group,γδT group,rapamycin treated γδT group and control group.Survival rate of MEC-1 cells in each group was determined by CCK-8 after 48 h of culture.Results:IL-12,IFN-γ and TNF-α levels secreted by γδT cells were the highest when rapamycin was 100 nmol/L,which was significantly higher than other groups(P<0.05).When rapamycin was 50 nmol/L,IL-17 secretion level of γδT cells was the highest,and IL-17 level secreted by γδT cells gradually decreased with increase of rapamycin concentration.Survival rates of MEC-1 cells in supernatant of DMSO group,rapamycin group,rapamycin treated γδT cell group and γδT cell group were(93.48±2.52)%,(71.44±4.31)%,(83.93±1.54)%and(57.97±2.47)%,whose differences were statistically significant compared with control group(P<0.05).Compared with control group,mTOR protein level of γδT cells was decreased in all rapamycin treated groups.Protein expressions of upstream AKT,pAKT and downstream specific transcription factor 4EBP-1 were decreased with increase of rapamycin concentration(P<0.05).Conclusion:There is a dose-effect relationship between rapamycin concentration and IL-17,IL-12,IFN-γ,TNF-α secretions by γδT cells.Rapamycin and γδT cells can inhibit pro-liferation of MEC-1 cells,and 100 nmol/L rapamycin can enhance inhibitory effect of γδT cells on proliferation of MEC-1 cells.Rapa-mycin can weaken AKT pathway on γδT cells,and rapamycin combined with γδT cells plays a positive role in anti-tumor application of leukemia.