Aim To examine the neuroprotective im-pacts of total saponins from Trillium tschonoskii maxim(TST)on cerebral ischemia-reperfusion injury(CIRI)in rats and delve into the mechanisms of ferroptosis.Methods The CIRI model was prepared by dividing male SD rats into the model group,TST(0.1 g·kg-1)group,Donepezil hydrochloride(0.45 mg·kg-1)group,and sham group.The cognitive functions of rats in each group were assessed through the Morris water maze test,the changes in neurological function were evaluated using the Zea-Longa method,the infarct area was observed via TTC staining,and the pathologi-cal alterations in brain tissue were analysed using HE and Nissl staining.To further investigate the underly-ing mechanism,the mitochondrial structural changes were examined using transmission electron microscopy,and the levels of GSH-PX,MDA,and SOD were ana-lyzed.Additionally,the expressions of GPX4 and Nrf2 proteins were evaluated through immunohistochemistry and immunofluorescence.Furthermore,the protein lev-els of Keap1/Nrf2/HO-1 and Nrf2/SLC7A11/GPX4 pathways in rats were examined using Western blot-ting.Results The rats in the model group displayed diminished learning and memory capabilities in com-parison to those in the sham group,as well as a signifi-cantly increased cerebral infarction area and higher neurological function scores(P＜0.01),significantly increased cerebral infarct area,disordered and loosely arranged neurons,and reduced Nissl bodies.Addition-ally,mitochondria showed typical signs of ferroptosis.Changes related to ferroptosis included decreased activ-ities of SOD and GSH-PX(P＜0.01)and increased MDA levels(P＜0.01).The expression of GPX4 and Nrf2-positive cells was significantly reduced,along with decreased fluorescence intensity of GPX4.Further-more,the protein expression of Keap1,Nrf2,HO-1,GPX4,SLC7A11 in the hippocampus decreased(P＜0.05,P＜0.01).Following the administration of TST,these effects showed improvement.Conclusions TST has neuroprotective effects,enhancing learning and memory abilities while reducing oxidative stress levels.The mechanism may involve the inhibition of ferroptosis through the Keap-1/Nrf2/HO-1 and Nrf2/SLC7 A11/GPX4 pathways.

Objective Mobile artificial lumbar complex(MALC)which suitable for reconstruction after subtotal lumbar resection in goats was developed,and to test stability of the complex and postoperative lumbar segmental motor function.Methods Eighteen male boer goats aged from 1 to 2 years old(weighted from 35 to 45 kg)were selected and divided into con-trol group,fusion group and non-fusion group,with 6 goats in each group.According to preoperative CT scans and MRI exami-nations of lumbar,the goat MALC was designed and performed by 3D printed for non-fusion group.Operation was performed on three groups respectively,and only vertebral body and disc were exposed in control group.In fusion group,L4 part of vertebral body and the upper and lower complete disc tissues were removed,and the lumbar spine bone plate fixation was performed with titanium mesh bone grafting.In non-fusion group,vertebral body and disc were removed in the same way,and MALC was im-planted.AP and lateral X-rays of lumbar vertebrae in goat were taken at 6 months after surgery,in order to understand whether the plant was dislocated,displaced and fractured.Biomechanical tests were performed on the specimens by mechanical instru-ment to measure range of motion(ROM)of L2,3,L,4,L4,5intervertebral space and the overall ROM of L2-5 lumbar vertebrae.Results MALC of lumbar vertebra was designed by 3D printing,and its component artificial vertebrae and upper and lower ar-tificial end plates were manufactured.The semi-spherical structure was fabricated by precision lathe using high-crosslinked polyethylene material,and the prosthesis was assembled.Postoperative AP and lateral X-rays of lumbar vertebra at 6 months showed the implant position of implant and MALC were good without displacement and dislocation.In vitro biomechanical test of lumbar vertebrae specimens:(1)There were no statistical significance in ROM of lumbar intervertebral space flexion and extension,lateral flexion and rotation on L.4 and L4,5,between non-fusion group and control group(P＞0.05),while ROM of fu-sion group was significantly reduced compared with the other two groups(P＜0.05).There were no significant difference in ROM of L2.3 intervertebral flexion and extension,lateral flexion and rotation between non-fusion group and control group(P＞0.05),while fusion group was significantly increased compared with the other two groups(P＜0.001).(2)There was no signifi-cant difference in overall lumbar ROM of L2-5(P＞0.05).Conclusion The individual MALC could restore intervertebral height of lumbar vertebra while maintaining the stability of lumbar vertebra and re-establishing motor function of lumbar space.

Objective To assess the clinical short-term outcomes of implanting D-shant atrial shunt device(aSD)in a single center for patients with heart failure with reduced ejection fraction(HFrEF).Methods From January 2022 to January 2023,a retrospective analysis was conducted on 12 patients with HFrEF who underwent percutaneous implantation of a D-shant aSD.We assessed cardiac chamber size and ventricular function using echocardiography,right heart catheterization measurements and patient clinical indicators were collected,follow up data of 12 months postoperative and pre-implantation D-shant were compared.The primary endpoint of the study was the cumulative occurrence of adverse cardiac,neurologic,or renal events during the follow-up period.Secondary endpoints were improvements in functional status included cardiac function,quality of life,and exercise capacity.Results All 12 patients underwent successful percutaneous inter-atrial shunting procedures using the D-shant.Postoperative immediately fluoroscopy and echocardiography confirmed accurate localization and patency of the atrial shunt devices in all cases.Postoperative hemodynamic assessment revealed a significant decrease in pulmonary capillary wedge pressure[(29.8±3.4)mmHg vs.(17.8±0.8)mmHg,P＜0.001].During 12 months follow-up,the cumulative adverse event rate was 8.3％(one patient received a heart transplant),a significant reduction in left atrial diameter from(65.8±6.5)mm to(48.0±4.5)mm(P＜0.001)was observed.Furthermore,there was notable improvement in clinical cardiac function indices quality of life,and exercise capacity of the patients.Conclusions This single-center retrospective study found that the use of a D-shant aSD to perform percutaneous interatrial shunting in patients with HFrEF is safe and effective.Short-term follow-up demonstrated sustained patency of the shunt and that the intervention was associated with improved functional status.

The effect of porcine SIRT5 on replication of foot and mouth disease virus type O(FMDV-O)and the underlying regulatory mechanism were investigated.Western blot and RT-qPCR analyses were employed to monitor expression of endoge-nous SIRT5 in PK-15 cells infected with FMDV-O.Three pairs of SIRT5-specific siRNAs were synthesized.Changes to SIRT5 and FMDV-O protein and transcript levels,in addition to virus copy numbers,were measured by western blot and RT-qPCR analyses.PK-15 cells were transfected with a eukaryotic SIRT5 expression plasmid.Western blot and RT-qPCR analyses were used to explore the impact of SIRT5 overexpression on FMDV-O replication.Meanwhile,RT-qPCR analysis was used to detect the effect of SIRT5 overexpression on the mRNA expression levels of type I interferon-stimulated genes induced by SeV and FMDV-O.The results showed that expression of SIRT5 was up-regulated in PK-15 cells infected with FMDV-O and siRNA interfered with SIRT5 to inhibit FMDV-O replication.SIRT5 overexpression promoted FMDV-O replication.SIRT5 over-expression decreased mRNA expression levels of interferon-stimulated genes induced by SeV and FMDV-O.These results suggest that FMDV-O infection stimulated expression of SIRT5 in PK-15 cells,while SIRT5 promoted FMDV-O rep-lication by inhibiting production of type I interferon-stimula-ted genes.These findings provide a reference to further ex-plore the mechanism underlying the ability of porcine SIRT5 to promote FMDV-O replication.

Heterotypic cell-in-cell structures(heCICs)mediate unique non-autonomous cell death,which are widely involved in a variety of important pathological processes,such as tumorigenesis,pro-gression and clinical prognosis.Methylenetetrahydrofolata dehydrogenase 2(MTHFD2),one of the key enzymes of one-carbon metabolism,is highly expressed in a variety of tumor cells.In this study,in order to investigate the effect of MTHFD2 on the formation of heCICs,liver cancer cells and immune cells were first labeled separately by live cell dyes,and the heCIC model was established by using fluorescence mi-croscopy for cell imaging and analysis.After transiently knocking down MTHFD2 in cells by RNAi,we found that the ability of PLC/PRF/5 and Hep3B to form heCICs with immune cells was significantly in-creased(all P＜0.01).MTHFD2 recombinant expression plasmid was constructed by the homologous re-combination method,and MTHFD2 overexpression cell lines were further constructed.Then,the effect of MTHFD2 overexpression on the ability to form heCICs was detected by co-culturing the overexpression cell lines with immune cells.The results showed that the rate of heCIC formation was significantly re-duced after overexpression of MTHFD2(all P＜0.001).In conclusion,this study demonstrated that MTHFD2 is a negative regulator of heCIC formation,providing a research basis for targeting MTHFD2 to promote heCIC formation and enhance the in-cell killing of immune cells.

Objective To establish a model of long-term atrazine(ATR)-induced liver injury in mice and to evaluate the hepatotoxic effects induced by ATR.Methods C57BL/6-N male mice were randomly divided into a control group and 1.5 mg/L and 150 mg/L ATR dose(ATR-L,ATR-H)groups.After 35 and 63 days,serum liver function biochemical indexes and inflammatory factors were detected,the hepatosomatic ratio was calculated,and the histopathology and ultrastructure of the liver were observed.Lipid peroxidation levels and antioxidant capacity,the activities of major phase I metabolic enzymes and phase Ⅱ detoxification enzymes,and the expression of related proteins in liver tissues were detected.Results Compared with the control group,the ATR groups showed significant changes in the AST/ALT ratio,levels of pro-inflammatory factors CCL2,TNF-α and IL-6,H2O2 content and activities of the metabolic enzymes NCR,CYTb5,and UDPGT(P＜0.05).In the 150 mg/L ATR group,GGT content,peroxide levels(as indicated by malondialdehyde),and CYP1A2 expression were significantly increased(P＜0.01),while GSH content was significantly decreased(P＜0.05),and hepatocyte injury and mitochondrial vacuolation were more serious when compared to control and 1.5 mg/L groups.Conclusions In a mouse model of low-dose ATR liver injury,both 1.5 mg/L and 150 mg/L ATR exposure induced liver injury in mice,with 150 mg/L ATR inducing the maximum metabolic toxicity in the liver after 63 days.

Objective:To investigate the efficacy and prognosis of chemotherapy regimen containing Bruton's tyrosine kinase(BTK)inhibitor in the treatment of relapsed/refractory mantle cell lymphoma(R/R MCL).Methods:The clinical data of 134 patients with R/R MCL were collected and analyzed retrospectively.The clinical characteristics of patients and effect of chemotherapy regimen on efficacy,overall survival(OS)and progression-free survival(PFS)were observed.Results:The median age of the patients was 58(56-61)years old,and male to female ratio was about 2.9∶1.Patients with Ann Arbor stage Ⅲ-Ⅳ accounted for 77.6％,extranodal involvement＞2 for 43.3％,bone marrow involvement for 60.4％,gastrointestinal involvement for 24.6％,and hepatosplenomegaly for 38.1％.The median follow-up time was 30(2-103)months,overall response rate(ORR)was 41.8％,3-year PFS was not reached,and 3-year and 5-year OS rate was 62.7％and 53.8％,respectively.The ORR of BTK inhibitor group was 56.9％,which was higher than 32.5％of non-BTK inhibitor group(P=0.006).The difference was statistically significant in PFS between the two groups(P=0.002),but was not in OS(P＞0.05).The difference was statistically significant in OS between classical and special morphology(P＜0.001),but was not in PFS(P＞0.05).Ki-67 was an influencing factor for OS and PFS.Multivariate analysis showed that Ki-67,B symptoms,MIPI score,and Ann Arbor stage were independent prognostic factors affecting patients'OS.The second-line treatment regimen was an independent prognostic factor affecting patients'PFS.Conclusions:The chemotherapy regimen containing BTK inhibitors can effectively improve the efficacy and prolong the PFS of R/R MCL patients.Ki-67,B symptoms,MIPI score,and Ann Arbor stage are independent prognostic factors for R/R MCL patients.

Objective:To explore the gene mutations of Langerhans cell histiocytosis in children,and to analyze the correlation of BRAF V600E mutation with clinical features and prognosis of LCH,so as to provide reference for clinical diagnosis and treatment. Methods:Fluorescence PCR was used to detect gene mutations in paraffin-embedded tissue samples from 78 children with LCH,and the correlation of BRAF V600E mutation with clinical characteristics and prognosis of LCH in children was analyzed. Results:Among the 78 children,41 cases (52.6％) had BRAF V600E mutation,8 cases (10.3％) had MAP2K1 mutation,1 case (1.3％) had BRAF Exon 12 mutation,1 case (1.3％) had ARAF mutation,and 1 case (1.3％) had PIK3CA mutation. BRAF V600E mutation was not significantly correlated with sex,age,multisystem involvement,risk-organ involvement,CNS-risk lesions,and early treatment response in children with LCH (P>0.05),and it was also not significantly correlated with the recurrence and event-free survival (EFS) of children with LCH (P>0.05). Conclusion:LCH is an inflammatory myeloid tumor. BRAF V600E mutation is not correlated with clinical features,early treatment response,recurrence and prognosis of LCH.

Objective:This paper aims to analyze the current status,characteristics,and problems of the policy text of China's chronic disease from the perspective of full-cycle,and provide a reference for subsequent policy optimization.Methods:The author selected 104 policy documents on chronic disease published at the national level from 2009 to 2023,used content analysis method,constructed a two-dimensional analysis framework of"policy tools-full-cycle management",and carried out one-dimensional quantitative analysis and two-dimensional cross-analysis.Results:The application of policy tools is obviously unbalanced,with supply-type policy tools accounting for as much as 63.23％,and demand-type policy tools used the least,only 10.51％.In the dimension of full-cycle management,the proportions of prevention,prevention and treatment,treatment,treatment and rehabilitation,health care,and full-cycle stages are 21.01％,24.58％,28.89％,7.32％,4.69％and 13.51％respectively.It is noteworthy that the proportions of the treatment and rehabilitation,and health care stages are relatively low.All stages of chronic disease health management interact frequently with supply-type policies,and the treatment and rehabilitation,health care and full-cycle stages cross less with demand-type policy tools.Conclusion:It is necessary to adjust the proportion of policy tools,strengthen their internal coordination,improve the top-level design of treatment and rehabilitation and health care,enhance the synergistic interaction between various types of policy tools and full-cycle management,and improve the overall effectiveness of policies.

Objective:To investigate the clinical and genetic characteristics of a male carrier of exceptional complex chromo-some rearrangement(CCR)and the outcome of preimplantation genetic testing for chromosomal structural rearrangement(PGT-SR).Methods:Using the modified high resolution G banding technique and whole-genome low-coverage sequencing(WGLCS),we analyzed the cellular karyotype and molecular karyotype of a male carrier of CCR,performed an analysis of the single-sperm chromosome copy number and conducted PGT-SR for the patient by next-generation sequencing(NGS).In addition,we reviewed the literature on repor-ted male carriers of CCRs and summarized their normal/balanced sperm ratios and PGT-SR outcomes.Results:The karyotype of the patient was 46,XY,der(5)inv(5)(q14.3q23.2)t(5;14;11)(q23.2;q31.1;q21),der(11)t(5;14;11);der(14)t(5;14;11),with the translocation breakpoints located in the intergenic region.Single-sperm sequencing revealed 20.0%(7/35)of normal haploids in the male's spermatozoa,and the results PGT-SR showed a proportion of 25.0%(4/16)of normal/balanced embryos.After thawing and transferring of 2 euploid blastocysts,a healthy male infant was successfully delivered.Conclusion:The proportion of normal hap-loids in the spermatozoa of male CCR carriers may be higher than theoretically predicted,and PGT-SR can effectively improve the preg-nancy outcome in male CCR carriers and provide valuable data for genetic counseling.