Pancreatic cancer is characterized by nerve invasion and a high mortality rate， and its pathological process depends on the complex interaction network between tumor and the nervous system. Based on the concept of “pancreatic cancer neuroecology”， this article analyzes the mechanism of action of peripheral motor nerve， sensory nerve， and central nerve in tumorigenesis， pain regulation， and cachexia formation and emphasizes the synergistic regulatory role of immune cells， Schwann cells， and extracellular matrix in the microenvironment of perineural invasion. At the same time， this article further elaborates on the metabolic interaction and chemotaxis between neuraxis and tumor， the effect on promoting chemotherapy resistance， and the dynamic relationship between neuroplasticity and tumor adaptability. In clinical practice， this article summarizes the key value of perineural invasion in prognostic evaluation， preoperative evaluation， and the selection of surgical strategy. In addition， this article reviews the basic research advances in the biomarkers and potential targets associated with perineural invasion in pancreatic cancer and points out the limitations of current model and transformation research. In the future， systematically analyzing the nerve-tumor-immune network and targeting its key nodes may provide multi-dimensional strategies and new breakthroughs for the precise intervention of pancreatic cancer， the reversal of drug resistance， and the relief of symptoms.

Objective: To investigate the effect of different volumes of isotonic washing solution (0.9% NaCl) on the quality of intraoperative autologous blood salvage in craniocerebral surgery. Methods: Thirty patients who underwent neurosurgical procedures with intraoperative salvaged blood volumes exceeding 800 mL between August 2022 and July 2024 were enrolled as study subjects. The salvaged blood was divided into four groups: group A, group B, group C, and group D. Group A was washed with 500 mL of isotonic solution (blood-to-washing solution ratio of 1∶2), group B with 1000 mL (ratio of 1∶4), and group C with 1 500 mL (ratio of 1∶6). After centrifugation, the washed blood was stored in designated blood bags (A, B, and C, respectively). Blood gas analysis was performed on 5 mL samples obtained from the centrifuged blood in groups A, B, and C, and from the unwashed blood in the reservoir of the control group (group D). Parameters including red blood cell (RBC) count, pH, hemoglobin (Hb), hematocrit (HCT), potassium ion (K ), sodium ion (Na ), and lactate levels were also measured using these samples. Additionally, blood samples from each group were examined microscopically to assess red blood cell morphology. Results: No significant differences were observed among groups A, B, and C in terms of RBC recovery rate, HCT, Na concentration, or lactate clearance rate (P>0.05). All three groups exhibited a reduction in pH, with group C showing a more pronounced decrease compared to groups A and B. Similarly, K clearance was achieved in all groups, with group C demonstrating superior efficiency. Microscopic analysis revealed a reduction in spiculated red blood cells across all groups, with group C achieving the highest clearance rate. Conclusion: Group C (washed with 1 500 mL solution) demonstrated optimal performance in reducing pH levels, clearing K , and removing spherocytes, while maintaining red blood cell recovery rate. Additionally, it did not significantly increase the risk of hypernatremia in critically ill neurosurgical patients, making it more suitable for craniocerebral surgeries.

Normal heart function depends on complex regulation by the brain, and abnormalities in the brain‒heart axis affect various diseases, such as myocardial infarction and anxiety disorders. However, systematic tracking of the brain regions associated with the input and output of the heart is lacking. In this study, we injected retrograde transsynaptic pseudorabies virus (PRV) and anterograde transsynaptic herpes simplex virus (HSV) into the left ventricular wall of mice to identify the whole-brain regions associated with the input to and output from the heart. We successfully detected PRV and HSV expression in at least 170 brain subregions in both male and female mice. Sex differences were discovered mainly in the hypothalamus and medulla, with male mice exhibiting greater correlation and hierarchical clustering than female mice, indicating reduced similarity and increased modularity of virus expression patterns in male mice. Further graph theory and multiple linear regression analysis of different injection timelines revealed that hub regions of PRV had highly similar clusters, with different brain levels, suggesting a top-down, hierarchically transmitted neural control pattern of the heart. Hub regions of HSV had scattered clusters, with brain regions gathered in the cortex and brainstem, suggesting a bottom-up, leapfrog, multipoint neural sensing pattern of the heart. Both patterns contain many hub brain regions that have been previously overlooked in brain‒heart axis studies. These results provide brain targets for future research and will lead to deeper insight into the brain mechanisms involved in specific heart conditions.
Animals ; Male ; Female ; Heart/physiology* ; Mice ; Herpesvirus 1, Suid ; Brain/physiology* ; Mice, Inbred C57BL ; Brain Mapping ; Efferent Pathways/physiology* ; Afferent Pathways/physiology* ; Simplexvirus ; Sex Characteristics

Objective To investigate the protective effect of exogenous leptin against focal cerebral ischemia-reperfusion(I/R)injury in mice and explore the underlying mechanism.Methods A total of 100 C57BL/6 mice were randomly divided into 5 groups,including a sham-operated group,cerebral I/R model group,and 3 leptin treatment groups with intraperitoneal injections of 0.5,1.0 or 2.0 leptin immediately after occlusion of the internal carotid artery.At 24 h after reperfusion,neurological function scores of the mice were assessed,and TTC staining was used to determine the area of cerebral infarction.The pathological changes in the cortical brain tissue of the mice were observed using HE staining,and degenerative damage of the cortical neurons were assessed with Fluoro-Jade C staining.The expression of glial fibrillary acidic protein in cortical brain tissues was detected using immunohistochemistry and Western blotting.In another 45 C57BL/6 mice with sham operation,I/R modeling,or leptin(1 mg/kg)treatment,glutamic acid in the cortical brain tissue was detected using glutamate assay,and cortical glutamate-aspartate transporter(GLAST)and glutamate transporter-1(GLT-1)protein expressions were detected using immunohistochemistry.Results Compared with the I/R model mice,the leptin-treated mice had significantly lower neurological deficit scores,smaller cerebral infarct area,milder pathologies in the cortical brain tissue,and lessened cortical neuronal damage with normal morphology and less excessive proliferation of the astrocytes.Leptin treatment significantly up-regulated the expressions of GLT-1 and GLAST and lowered the content of glutamic acid in the brain tissue of the I/R mice.Conclusion Exogenous leptin has obvious neuroprotective effect against cerebral I/R injury in mice,mediated probably by controlling excessive astrocyte proliferation and up-regulating cortical GLT-1 and GLAST expressions to reduce glutamate-mediated excitotoxic injury of the astrocytes.

Objective To investigate the mechanism of the interaction between miR-9-5p and tissue metalloproteinase inhibitor 2(TIMP2)on autophagy and apoptosis in multiple myeloma(MM)cells.Methods Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect expression levels of miR-9-5p and TIMP2 in bone marrow samples of 9 patients with newly diagnosed MM and 9 patients with recurrent MM.The correlation of expression levels between the two were analyzed.U266 cells were divided into the miR-control group,the miR-9-5p group,the pcDNA3.1 group,the pcDNA3.1-TIMP2 group,the miR-9-5p+pcDNA3.1 group,and the miR-9-5p+pcDNA3.1-TIMP2 group.The effects of overexpressed miR-9-5p and TIMP2 on autophagy and apoptosis in U266 cells were detected by flow cytometry,immunofluorescence staining and Western blot experiments.The dual luciferase report experiment verified the interaction between miR-9-5p and TIMP2.Results Compared with newly diagnosed MM patients,the expression level of miR-9-5p was increased and the expression level of TIMP2 was decreased in patients with recurrent MM.The expression levels of miR-9-5p and TIMP2 were negatively correlated(P<0.05).Compared with the miR-control group,the miR-9-5p group showed a decrease in the expression level of MAP1LC3B-Ⅱ,an increase in expression levels of MAP1LC3B-Ⅰ and SQSTM1,and a decrease in cell apoptosis rate(P<0.05).Compared with the pcDNA3.1 group,the expression level of MAP1LC3B-Ⅱ was increased in the pcDNA3.1-TIMP2 group,while the expression levels of MAP1LC3B-Ⅰ and SQSTM1 were decreased,and the apoptosis rate of cells increased(P<0.05).Bioinformatics and dual luciferase reporter experiments confirmed that TIMP2 was the target gene of miR-9-5p.Conclusion miR-9-5p inhibits autophagy and apoptosis in MM cells by targeting TIMP2,thereby promoting the occurrence and development of MM.

Objective:To screen ferroptosis genes related to prognosis of malignant pleural mesothelioma(MPM),explore the relationship between ferroptosis and tumor immune microenvironment and provide a new perspective for targeting and immunotherapy of MPM patients.Methods:The differentially expressed genes(DEGs)in MPM tumor group and normal group were analyzed in GEO database;intersection of DEGs and ferroptosis genes to obtain differentially expressed ferroptosis-related genes(DE-FRGs).GO,KEGG function enrichment and protein protein interaction(PPI)were used to identify the signal pathways mainly involved by DE-FRGs.The prognosis related ferroptosis genes were identified by univariate COX analysis.LASSO regression analysis was used to screen the best DE-FRGs for establishing the risk prediction model,and a risk prognosis model based on the best DE-FRGs was estab-lished by multivariate cox analysis to verify the prediction effect of the model.Finally,CIBERSORT and other algorithms were used to analyze tumor immune cell infiltration and evaluate immune microenvironment.Results:Twenty-four prognosis related DE-FRGs were screened,which were mainly concentrated in ferroptosis,transcriptional regulation and response to inorganic substances.A MPM risk prediction model based on five ferroptosis-related genes(ALDH3A2,CAV1,HRAS,CDCA3 and RRM2)was established and vali-dated.In the model,the proportion of CD8+T cells and macrophages in high-risk group were higher,while the proportion of B lympho-cytes was lower.In addition,PD-1,CTLA-4 and their ligands at immune checkpoint had higher expression status in high-risk group.Conclusion:The MPM risk prediction model based on five ferroptosis-related genes is established,and the immune status in the model is clarified.It provides a certain research basis for targeting and immunotherapy of MPM.The predictive ability of this model in MPM needs to be further verified in clinical practice to better predict disease stratification and treatment management.

Objective To explore the mechanism of ginsenoside Rg1 in alleviating heat stress-induced testicular injury in mice.Methods A total of 20 C57BL/6 male mice(6～8 weeks old)were randomly divided into 4 groups(n=5).The mice from the control group and heat stress(HS)group were intraperitoneally injected with 10 mL/(kg·d)0.9％normal saline for 14 d,while those in the HS+Rg1 group and the Rg1 group were given an intraperitoneal injection of 20 mg/(kg·d)for 14 d,and then on the 7th day after administration,the mice in the HS group and the HS+Rg1 group had the lower abdomen put into a 43 ℃ water bath for 30 min as a single heat stress after being anesthetized with 4％chloral hydrate.Mouse spermatocytes GC-2spd(ts)were divided into control group(routine culture for 48 h),HS group(placed in a 43 ℃ water bath for 30 min after 36 h of conventional culture,and cultured till the end of 48 h),HS+Rg1 group(50 μmol/L Rg1 treatment followed by heat stress injury),and Rg1 group(no heat stress injury).In 1 d after modeling,the eyeball blood samples were collected to detect serum testosterone with ELISA,and the testicles were extracted to observe the morphology and weighed to calculate the testicular index.HE staining was used to observe the histopathology of testis,and corresponding reagents and kits was employed to detect the content of malondialdehyde(MDA)and activities of catalase(CAT)and superoxide dismutase(SOD)in testis tissue.After the epididymal sperm were collected,the sperm concentration and motility were analyzed by computer-assisted sperm analysis(CASA)system.In in vitro experiments,cell apoptosis was detected with TUNEL staining,the protein levels of Nrf2,Keap1,HO-1,Bax,Bcl-2 and Caspase3 were detected with Western blotting,and the mRNA levels of GCLC,GCLM and NQO1 were detected by RT-qPCR.Results Rg1 prevented the decreases in testicular weight and testicular index caused by heat stress,reduced the damage of testicular tissue structure,prevented the decrease of sperm concentration and vitality,antagonized the decreasd number of Leydig cells and serum testosterone level,reduced the accumulation of MDA in testicular tissue,and enhanced the activities of CAT and SOD.Rg1 treatment alleviated the apoptosis of GC-2spd(ts)cells,down-regulated the expression of Bax,Caspase3 and Keap1 proteins,enhanced the expression of Bcl-2,Nrf2 and HO-1 proteins,and increased the transcriptional levels of Nrf2 target genes GCLC,GCLM and NQO1.Conclusion Rg1 has no significant effect on the structure and function of mouse testes,but it can effectively improve the ability of mouse testes to resist heat stress injury,which may be related to the activation of Nrf2 signaling pathway,the improvement of antioxidant enzyme activity,and the reduction of apoptosis of spermatogenic cells.

Objective To make transfusion management strategies for patients with history of blood transfusion and/or pregnancy by following up a patient with delayed hemolytic transfusion reactions(DHTR)caused by unexpected antibody produced after blood transfusion.Methods ABO,Rh,MN and Kidd blood group test,direct antiglobulin test,unexpected antibody screening,antibody identification,antibody titer detection,and cross-matching test were performed on a patient with DHTR.Meanwhile,suitable red blood cells were screened for subsequent treatment.Results The patient′s blood group was B,RhD(+)and CCDee,the antibody screening test and cross-matching test were negative before the first trans-fusion.After eight days,hemoglobin of the patient decreased to 57 g/L and the laboratory results indicated delayed hemoly-sis,the antibody screening was positive,and the antibody identification result was anti-M,as RBCs of the patient received typed as M+N+.After the patient received M antigen negative RBCs,the laboratory test results still indicated delayed sero-logic transfusion reaction.A new antibody arose and was identified as anti-Jka while RBCs transfused were M-N+and Jk(a+b-).Afterwards,it was effective for the patient to receive B,RhD(+),M-N+and Jk(a-b+)RBCs.Conclusion Most of the homologous antibodies produced by patients after blood transfusion will disappear within a few years.When patients undergo another transfusion,DHTR may occur because of anamnestic reaction.Establishing a transfusion management docu-ment and creating a card for patients who have already produced RBC alloantibodies can greatly reduce the occurrence of DHTR by informing doctors and staff when the next transfusion is needed.

Objective To investigate the protective effect of exogenous leptin against focal cerebral ischemia-reperfusion(I/R)injury in mice and explore the underlying mechanism.Methods A total of 100 C57BL/6 mice were randomly divided into 5 groups,including a sham-operated group,cerebral I/R model group,and 3 leptin treatment groups with intraperitoneal injections of 0.5,1.0 or 2.0 leptin immediately after occlusion of the internal carotid artery.At 24 h after reperfusion,neurological function scores of the mice were assessed,and TTC staining was used to determine the area of cerebral infarction.The pathological changes in the cortical brain tissue of the mice were observed using HE staining,and degenerative damage of the cortical neurons were assessed with Fluoro-Jade C staining.The expression of glial fibrillary acidic protein in cortical brain tissues was detected using immunohistochemistry and Western blotting.In another 45 C57BL/6 mice with sham operation,I/R modeling,or leptin(1 mg/kg)treatment,glutamic acid in the cortical brain tissue was detected using glutamate assay,and cortical glutamate-aspartate transporter(GLAST)and glutamate transporter-1(GLT-1)protein expressions were detected using immunohistochemistry.Results Compared with the I/R model mice,the leptin-treated mice had significantly lower neurological deficit scores,smaller cerebral infarct area,milder pathologies in the cortical brain tissue,and lessened cortical neuronal damage with normal morphology and less excessive proliferation of the astrocytes.Leptin treatment significantly up-regulated the expressions of GLT-1 and GLAST and lowered the content of glutamic acid in the brain tissue of the I/R mice.Conclusion Exogenous leptin has obvious neuroprotective effect against cerebral I/R injury in mice,mediated probably by controlling excessive astrocyte proliferation and up-regulating cortical GLT-1 and GLAST expressions to reduce glutamate-mediated excitotoxic injury of the astrocytes.