Objective:To investigate the effect of Lulongzaisheng decoction on ferroptosis in aplastic anemia (AA) model mice and its potential mechanism of action on AA.Methods:Using female BALB/c mice as controls (control group), a mouse model of AA was established by intraperitoneal injection of interferon-γ (IFN-γ) combined with oral gavage of busulfan. The AA group was treated with continuous oral gavage of Lulongzaisheng decoction for 10 days. The effects on the Jak2/Stat3/Acsl4 signaling pathway were assessed. Bone marrow biopsy and histopathological examination were performed to observe the degree of AA in each group of mice. Flow cytometry was used to detect the production of lipid reactive oxygen species (ROS) in the bone marrow cells. Immunohistochemical staining was performed to observe the expression of 4-HNE in the sternum. Western blot analysis was conducted to determine the protein expression levels of Jak2, p-Jak2, Stat3, p-Stat3, and Acsl4. ELISA was used to measure the levels of interleukin-6 (IL-6) in the peripheral blood plasma.Results:Compared with the control group, mice in the AA group exhibited a reduction in hematopoietic tissues within the bone marrow cavity and significant fatty infiltration. In contrast, mice in the AA+Lulongzaisheng decoction group showed partial recovery of the bone marrow and a decrease in fatty infiltration. The lipid ROS proportions in the control, AA, and AA+Lulongzaisheng decoction groups were (47.01±3.07) %, (53.81±1.99) %, and (49.50±3.98) %, respectively ( P<0.05), indicating that the AA + Lulongzaisheng decoction group had a slightly lower accumulation of lipid ROS than the AA group. The expression areas of 4-HNE in the bone marrow cavity for the control, AA, and AA+Lulongzaisheng decoction groups were (6.34±1.07) %, (35.26±3.68) %, and (16.97±1.30) %, respectively ( P<0.05), demonstrating a significant reduction in 4-HNE accumulation in the bone marrow in the AA + Lulongzaisheng decoction group compared with that in the AA group. Compared with the control group, the AA group exhibited upregulated expression of the ferroptosis-related protein Acsl4, along with increased expression of p-Stat3 and p-Jak2. In the AA+ Lulongzaisheng decoction group, the expression of Acsl4 was downregulated, whereas those of p-Stat3 and p-Jak2 were inhibited ( P<0.05). The levels of IL-6 in the peripheral blood plasma were (65.60±6.01), (166.50±3.32), and (119.37±4.29) pg/ml in the control, AA, and AA+Lulongzaisheng decoction groups, respectively ( P<0.05). Compared with the AA group, the AA + Lulongzaisheng decoction group exhibited a significant decrease in IL-6 levels. Conclusion:This study shows that Lulongzaisheng decoction has a significant therapeutic effect on AA model mice by regulating the Jak2/Stat3/Acsl4 pathway. The mechanism of action of this traditional Chinese medicine involves inhibiting the Jak2/Stat3 signaling pathway and regulating the expression of Acsl4, thereby improving hematopoietic function in AA model mice. These findings provide new drug targets and treatment strategies for the treatment of AA.

Objective:To evaluate the impact of donor characteristics on the prognosis of myelodysplastic syndrome (MDS) patients undergoing haplo-identical transplantation (HIDT) .Methods:A retrospective analysis of 203 MDS patients who received HIDT was conducted to evaluate how donor factors influenced transplant outcomes.Results:In MDS patients undergoing haploidentical transplantation, donors over 50 years were associated with higher EBV reactivation (2-year cumulative incidence 42.9% vs 22.0% for <50 years old; P=0.010). Female donors were linked to increased severe chronic GVHD compared with male donors (2-year incidence 11.9% vs 4.0% ; P=0.017). Additionally, 2-year overall survival (OS) was slightly lower with female donors than male donors (56.6% vs 69.7% ), but the difference was not statistically significant ( P=0.073). Donor-recipient blood type did not affect post-transplant OS or cumulative relapse rates. Donor-recipient kinship analysis revealed that child donors, compared to haploidentical sibling or parent donors, had lower rates of grade Ⅱ–Ⅳ acute GVHD (27.2% vs 45.7% vs 53.5%, P=0.007) and 2-year EBV reactivation (13.9% vs 29.3% vs 38.9%, P=0.001). For donors under 20 years, donor gender did not significantly affect 2-year OS ( P=0.913), relapse-free survival ( P=0.716), or 100-day incidence of grade Ⅱ–Ⅳ acute GVHD ( P=0.359) . Conclusion:For MDS patients undergoing HIDT, donors over 50 should be avoided. Male and child donors are preferred, while donor gender does not significantly affect outcomes if the donor is under 20 years old.

Primary splenic lymphoma is a rare malignant neoplasm， with similar clinical manifestations to Sjogren’s syndrome and liver cirrhosis， which often leads to misdiagnosis. This article reports a case of primary splenic lymphoma misdiagnosed as Sjogren’s syndrome with liver cirrhosis， in order to improve the understanding of primary splenic lymphoma， Sjogren’s syndrome， and liver cirrhosis and avoid misdiagnosis and treatment delay.

Objective:To explore the efficacy and safety of orelabrutinib combined with R-CHOP in patients with high-risk nongerminal center B-cell (non-GCB) diffuse large B-cell lymphoma (DLBCL) with extranodal involvement.Methods:This retrospective study was conducted on 35 patients who were seen at Guangxi Medical University Cancer Hospital and were immunohistochemically confirmed to have non-GCB DLBCL, had an International Prognostic Index score of 3 - 5, and confirmed to have ≥2 extranodal involvement on PET/CT. The treatment comprised the standard R-CHOP regimen combined with oral orelabrutinib (150 mg/day) for six cycles. In patients who developed neutropenia or grade 3 neutropenia with fever during treatment, administration of prophylactic pegylated granulocyte colony-stimulating factor 48 h after the end of chemotherapy was started on the next cycle. The endpoints included overall response rate (ORR), complete response (CR) rate, progression-free survival (PFS) time, overall survival (OS) time, and safety assessment.Results:The 35 eligible patients enrolled had a median age of 53 years (21 - 72 years) and a median follow-up time of 28 months (12 - 36 months) ; 19 patients had double-expressor (DE) status. The ORR was 88.6%, and the CR rate was 68.6%. The 2-year PFS and OS rates were 68.6% (95% CI 54.0% - 7.2%) and 87.5% (95% CI 76.7% - 100%), respectively. The 2-year PFS rate was significantly lower in patients with DE status than in those without DE status [54.4% (95% CI 35.4% - 84.2%) vs. 85.2% (95% CI 68.3% - 100%), P=0.048]. Serious adverse events included febrile neutropenia, pneumonia, and atrial flutter, but no treatment-related deaths. Conclusion:In patients with high-risk non-GCB DLBCL and extranodal involvement, the combination of orelabrutinib with R-CHOP regimen had good efficacy and manageable toxicity.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

Objective:To explore the diagnostic key points, treatment strategies, and prognosis of graft-versus-host disease (GVHD) after liver transplantation.Methods:The clinical data of 5 recipients diagnosed with GVHD after liver transplantation at the Liver Transplantation Center of the First Affiliated Hospital of Army Medical University from May 1999 to October 2024 were retrospectively analyzed. The causes, onset, diagnosis, treatment, and prognosis of GVHD after liver transplantation were summarized and analyzed. Literature was searched in CNKI, Wanfang, VIP, Chinese Medical Journal Full-text Database, PubMed, Web of Science, and Google Scholar using the Chinese keywords "移植物抗宿主病+肝移植", and the English keywords "graft versus host disease + liver transplantation". The search time ranged from January 1988 to January 2025. Inclusion criteria for the literature: (1) meeting the clinical or pathological diagnostic criteria of GVHD after liver transplantation; (2) recipient age >18 years; (3) case number ≥2. Exclusion criteria: incomplete clinical data such as incidence, mortality, and clinical manifestations of GVHD after liver transplantation. The retrieved literature was reviewed.Results:All 5 recipients were male. Among them, 4 cases underwent liver transplantation at this center. The incidence of GVHD after liver transplantation in this center was 0.46% (4/872). All 5 cases developed symptoms such as fever, rash, diarrhea, oral ulcers, and pancytopenia on the 19th (5-21) day after liver transplantation. One case had gastrointestinal bleeding. Two cases were diagnosed by skin pathological biopsy, and three cases were diagnosed based on clinical manifestations such as fever, rash, diarrhea, and bone marrow suppression. One case discontinued immunosuppressants, and four cases reduced the dosage of immunosuppressants. Four cases were treated with high-dose glucocorticoids, four with intravenous immunoglobulin (IVIG), three with ruxolitinib, and three with hematopoietic factors. All five cases received protective isolation, anti-infection, and symptomatic supportive treatment. Among the three recipients treated with ruxolitinib, body temperature returned to normal, rash gradually faded, oral ulcers gradually healed, blood cells returned to normal, and they were eventually discharged after recovery. The remaining two cases showed no symptom improvement and died of severe lung infection and multiple organ failure. Literature review A total of 34 articles were included. The incidence of GVHD after liver transplantation was 1.03% (279/27 018), and the onset time ranged from 7 to 1,865 days post-transplantation; 272 cases (97.49%) occurred within 1-8 weeks. The main clinical manifestations included fever (195 cases, 69.89%), rash (267 cases, 95.70%), diarrhea (173 cases, 62.01%), and bone marrow suppression (214 cases, 76.70%). Treatment mainly involved adjustment of immunosuppressants (201 cases, 72.04%), high-dose corticosteroids (215 cases, 77.06%), and IVIG pulse therapy (146 cases, 52.33%). In the end, 83 cases (29.75%) recovered and were discharged, while the mortality rate was 70.25% (196/279), with causes of death including infection, gastrointestinal bleeding, and multiple organ failure.Conclusions:GVHD after liver transplantation has a low incidence, high mortality, and poor prognosis. Diagnosis mainly relies on typical clinical manifestations and pathological results of tissue biopsy. Early administration of high-dose corticosteroids combined with IVIG pulse therapy, timely reduction or discontinuation of immunosuppressants, use of ruxolitinib, active infection management, and enhanced symptomatic and supportive care are effective strategies for treating GVHD after liver transplantation.

Objective To explore the interplay between MCT1-mediated lactate accumulation and ferroptosis in acute liver failure(ALF).Methods An ALF mouse model and a hepatocyte injury model were established using lipopolysaccharide(LPS)in combination with D-galactosamine(D-GalN).The mice were randomly assigned to three groups:a blank control group,an ALF model group,and an ALF+Liproxstatin-1(Lip-1)treatment group.In vitro experiments included four groups:A blank control,a hepatocyte injury model,a lactate intervention,and an MCT1 overexpression group.Commercial kits were used to measure lactate levels in both mouse liver tissues and cell supernatants,as well as the contents of malondialdehyde(MDA),ferrous ions(Fe2+),and reduced glutathi-one(GSH)in liver tissue.Liver histopathology was evaluated using hematoxylin and eosin(HE)staining.Trans-mission electron microscopy was employed to assess mitochondrial ultrastructure in hepatocytes.Western blot(WB)analysis was performed to determine the protein expression levels of MCT1,glutathione peroxidase 4(GPX4),and acyl-CoA synthetase long-chain family member 4(ACSL4)in both liver tissues and cultured cells.Real-time quantitative PCR and immunofluorescence assays were utilized to detect mRNA expression and fluorescence intensity of GPX4 and ACSL4,respectively.Results HE staining of liver tissue from the ALF mouse model revealed extensive hepatocyte necrosis and partial inflammatory cell infiltration.Both MCT1 and GPX4 protein expression were significantly downregulated(P<0.001),whereas ACSL4 protein expression was markedly upregulated(P<0.000 1),accompanied by a significant elevation in lactate levels(P<0.001).Trans-mission electron microscopy demonstrated reduced mitochondrial volume and disorganized cristae arrangement in hepatocytes.In contrast to the model group,histological analysis of liver tissue from ALF mice treated with an iron death inhibitor showed attenuated liver injury.GPX4 expression was restored(P<0.05),while ACSL4 expression was reduced(P<0.001).Levels of lactate,MDA,and Fe2+in liver tissue were significantly lower(P<0.001),whereas GSH levels were significantly higher(P<0.05).In vitro experiments indicated that lactate treatment suppressed GPX4 expression in hepatocytes in a concentration-dependent manner while promoting ACSL4 expres-sion(P<0.05).In the hepatocyte injury model group,MCT1 and GPX4 expression were downregulated,ACSL4 protein expression was upregulated(P<0.05),and lactate levels were significantly increased(P<0.05).However,MCT1 overexpression effectively reversed these alterations,resulting in increased GPX4 expression(P<0.05)and decreased ACSL4 expression(P<0.001).Furthermore,immunofluorescence results revealed enhanced GPX4 fluorescence intensity(P<0.001)and reduced ACSL4 signal intensity(P<0.01),along with a marked reduction in lactate levels in cell supernatants(P<0.000 1).Conclusions This study demonstrates that ferroptosis plays a critical role in cell death during ALF and is closely intertwined with lactate metabolism.MCT1 mitigates LPS/D-GalN-induced ferroptosis by facilitating lactate transport in hepatocytes,thereby reducing lactate accumulation.Conversely,inhibition of ferroptosis leads to decreased lactate levels,indicating a bidirectional'lactate-ferroptosis'regulatory loop.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

Objective To explore the interplay between MCT1-mediated lactate accumulation and ferroptosis in acute liver failure(ALF).Methods An ALF mouse model and a hepatocyte injury model were established using lipopolysaccharide(LPS)in combination with D-galactosamine(D-GalN).The mice were randomly assigned to three groups:a blank control group,an ALF model group,and an ALF+Liproxstatin-1(Lip-1)treatment group.In vitro experiments included four groups:A blank control,a hepatocyte injury model,a lactate intervention,and an MCT1 overexpression group.Commercial kits were used to measure lactate levels in both mouse liver tissues and cell supernatants,as well as the contents of malondialdehyde(MDA),ferrous ions(Fe2+),and reduced glutathi-one(GSH)in liver tissue.Liver histopathology was evaluated using hematoxylin and eosin(HE)staining.Trans-mission electron microscopy was employed to assess mitochondrial ultrastructure in hepatocytes.Western blot(WB)analysis was performed to determine the protein expression levels of MCT1,glutathione peroxidase 4(GPX4),and acyl-CoA synthetase long-chain family member 4(ACSL4)in both liver tissues and cultured cells.Real-time quantitative PCR and immunofluorescence assays were utilized to detect mRNA expression and fluorescence intensity of GPX4 and ACSL4,respectively.Results HE staining of liver tissue from the ALF mouse model revealed extensive hepatocyte necrosis and partial inflammatory cell infiltration.Both MCT1 and GPX4 protein expression were significantly downregulated(P<0.001),whereas ACSL4 protein expression was markedly upregulated(P<0.000 1),accompanied by a significant elevation in lactate levels(P<0.001).Trans-mission electron microscopy demonstrated reduced mitochondrial volume and disorganized cristae arrangement in hepatocytes.In contrast to the model group,histological analysis of liver tissue from ALF mice treated with an iron death inhibitor showed attenuated liver injury.GPX4 expression was restored(P<0.05),while ACSL4 expression was reduced(P<0.001).Levels of lactate,MDA,and Fe2+in liver tissue were significantly lower(P<0.001),whereas GSH levels were significantly higher(P<0.05).In vitro experiments indicated that lactate treatment suppressed GPX4 expression in hepatocytes in a concentration-dependent manner while promoting ACSL4 expres-sion(P<0.05).In the hepatocyte injury model group,MCT1 and GPX4 expression were downregulated,ACSL4 protein expression was upregulated(P<0.05),and lactate levels were significantly increased(P<0.05).However,MCT1 overexpression effectively reversed these alterations,resulting in increased GPX4 expression(P<0.05)and decreased ACSL4 expression(P<0.001).Furthermore,immunofluorescence results revealed enhanced GPX4 fluorescence intensity(P<0.001)and reduced ACSL4 signal intensity(P<0.01),along with a marked reduction in lactate levels in cell supernatants(P<0.000 1).Conclusions This study demonstrates that ferroptosis plays a critical role in cell death during ALF and is closely intertwined with lactate metabolism.MCT1 mitigates LPS/D-GalN-induced ferroptosis by facilitating lactate transport in hepatocytes,thereby reducing lactate accumulation.Conversely,inhibition of ferroptosis leads to decreased lactate levels,indicating a bidirectional'lactate-ferroptosis'regulatory loop.

Objective:To explore the efficacy and safety of orelabrutinib combined with R-CHOP in patients with high-risk nongerminal center B-cell (non-GCB) diffuse large B-cell lymphoma (DLBCL) with extranodal involvement.Methods:This retrospective study was conducted on 35 patients who were seen at Guangxi Medical University Cancer Hospital and were immunohistochemically confirmed to have non-GCB DLBCL, had an International Prognostic Index score of 3 - 5, and confirmed to have ≥2 extranodal involvement on PET/CT. The treatment comprised the standard R-CHOP regimen combined with oral orelabrutinib (150 mg/day) for six cycles. In patients who developed neutropenia or grade 3 neutropenia with fever during treatment, administration of prophylactic pegylated granulocyte colony-stimulating factor 48 h after the end of chemotherapy was started on the next cycle. The endpoints included overall response rate (ORR), complete response (CR) rate, progression-free survival (PFS) time, overall survival (OS) time, and safety assessment.Results:The 35 eligible patients enrolled had a median age of 53 years (21 - 72 years) and a median follow-up time of 28 months (12 - 36 months) ; 19 patients had double-expressor (DE) status. The ORR was 88.6%, and the CR rate was 68.6%. The 2-year PFS and OS rates were 68.6% (95% CI 54.0% - 7.2%) and 87.5% (95% CI 76.7% - 100%), respectively. The 2-year PFS rate was significantly lower in patients with DE status than in those without DE status [54.4% (95% CI 35.4% - 84.2%) vs. 85.2% (95% CI 68.3% - 100%), P=0.048]. Serious adverse events included febrile neutropenia, pneumonia, and atrial flutter, but no treatment-related deaths. Conclusion:In patients with high-risk non-GCB DLBCL and extranodal involvement, the combination of orelabrutinib with R-CHOP regimen had good efficacy and manageable toxicity.