BACKGROUND:Growth factor is the key effect molecule that plays a role in platelet-rich plasma in clinical treatment.There are differences in the concentration of growth factor after different activators activate platelet-rich plasma,which is an important factor affecting clinical efficacy. OBJECTIVE:To analyze the influence of different activators on the mass concentration of growth factors in platelet-rich plasma. METHODS:Totally 12 healthy volunteers were recruited to collect EDTA-K2 anticoagulant venous blood.Secondary centrifugation was used to prepare platelet-rich plasma.The difference in mass concentrations of growth factors was compared between venous blood and platelet-rich plasma.The platelet-rich plasma was mixed with four activators(normal saline,thrombin,calcium gluconate,calcium gluconate+thrombin)according to the volume ratio of 10:1,and incubated in a constant temperature water bath at 37 °C for 30 minutes.After centrifugation,the supernatant was extracted and the mass concentration of growth factor was detected.The bacterial growth in supernatant was measured by blood agar plate.Pearson correlation was used to analyze the correlation between different activators and the mass concentration of growth factor in platelet-rich plasma,and the correlation between the value of thrombocytometer and the mass concentration of growth factors in platelet-rich plasma. RESULTS AND CONCLUSION:(1)The mass concentrations of platelet-derived growth factor-BB,platelet-derived growth factor-AB,vascular endothelial growth factor,and epidermal growth factor in platelet-rich plasma were 8.7,22.2,2.3,and 2.8 times of those in venous blood,respectively(P<0.05).(2)Compared with normal saline group,the mass concentrations of platelet-derived growth factor BB,platelet-derived growth factor AB,vascular endothelial growth factor,and epidermal growth factor were increased in the thrombin group,calcium gluconate group,and calcium gluconate+thrombin group(P<0.05).The mass concentration of platelet-derived growth factor BB in the thrombin group and calcium gluconate group was higher than that in the calcium gluconate+thrombin group(P<0.05),and the mass concentration of platelet-derived growth factor AB in the thrombin group was higher than that in the calcium gluconate group and calcium gluconate+thrombin group(P<0.05).Epidermal growth factor mass concentration in the thrombin group was lower than that in the calcium gluconate group and calcium gluconate+thrombin group(P<0.05).(3)The results of blood agar plate test showed no bacterial growth in the supernatant of the four groups.(4)Pearson correlation analysis showed that the mass concentration of platelet-derived growth factor BB in platelet-rich plasma was strongly positively correlated with thrombin(r=0.683,P<0.05),and the mass concentration of vascular endothelial growth factor was strongly positively correlated with thrombin,calcium gluconate,calcium gluconate+thrombin stimulant(r=0.730,0.789,0.686,P<0.05).There was no correlation between the value of thrombocytometer and the mass concentration of four kinds of growth factors(P>0.05).(5)The results suggest that different activators have an impact on the concentration of growth factors in platelet-rich plasma.It is suggested to choose different activators to improve clinical efficacy according to different growth factor mass concentrations and treatment needs.

Based on his extensive clinical experience and the team's mechanistic research, Professor XU Nenggui has proposed the academic concept that "the governor vessel governs the brain and treats cerebral viscus diseases", and established a novel acupuncture approach for encephalopathy treatment centered on the integrated theory of "governor vessel-brain-mind", and developed a staged acupuncture protocol of "governor vessel regulating spirit" for ischemic stroke. This article introduces the academic features of this method in treating post-stroke dysphagia from four aspects: theoretical framework, treatment principles and point selection, mechanistic research, and clinical case studies. In clinical application, the method emphasizes syndrome differentiation based on meridians, harmonization between the conception and governor vessels; precise acupoint selection to treat both form and spirit; stage-specific differentiation with targeted needling. Furthermore, Professor XU integrates basic research with clinical practice, focusing on the neurobiological mechanisms underlying the efficacy of acupuncture in treating post-stroke dysphagia.
Acupuncture Therapy/methods* ; Humans ; Deglutition Disorders/psychology* ; Stroke/complications* ; Meridians ; Acupuncture Points

BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

Objective Using Meta-analysis to systematically evaluate the incidence and risk factors of pulmonary infection occurring in patients with acute ischemic stroke(AIS)after receiving mechanical thrombectomy.Methods A computerized retrieval of academic papers concerning the case-control study and cohort study of lung infection in patients with AIS after mechanical thrombectomy from the databases of PubMed,Web of Science,Embase,Cochrane Library,China Knowledge Network,Wanfang Data Knowledge Service Platform,and Wipro.com databases was conducted by two independent investigators.The retrieval time period was from the establishment of the database to December 2023.STAT A 16.0 software was used to make the Meta-analysis of the extracted literature.Results A total of 10 literature were included,with a combined sample size of 6 149 cases and 2 076 cases occurring lung infections.Meta-analysis revealed that the pulmonary infection in patients after mechanical thrombectomy for AIS showed a high degree of heterogeneity in the included studies(I2=97.87％,P＜0.001),and a random-effects model yielded an incidence of 31.8％for pulmonary infection.Risk factors for the occurrence of postoperative lung infection included atrial fibrillation(OR=1.33,95％CI:1.10-1.6,P=0.004),dysphagia(OR=3.83,95％CI:2.43-6.05,P＜0.001),high NIHSS score(OR=1.43,95％CI:1.05-1.94,P=0.023),impaired consciousness(OR=2.29,95％CI:1.38-3.81,P＜0.001),and intraoperative use of sedative drugs(OR=3.21,95％CI:2.91-3.54,P＜0.001).Conclusion The existing evidences indicate that patient's age ≥60 years,atrial fibrillation,dysphagia,impaired consciousness,high NIHSS score,and intraoperative use of sedative drugs are risk factors for the occurrence of pulmonary infection in AIS patients after receiving mechanical thrombectomy.In clinical practice,these risk factors should be highly valued,and these risk factors should be included in the postoperative comprehensive assessment and intervention strategies for patients so as to reduce the incidence of pulmonary infection.

Objective To evaluate the risk factors for pulmonary infection in patients with acute ischemic stroke after receiving mechanical thrombectomy and to construct a risk prediction model based on machine-learning algorithm technology.Methods A total of 315 patients with acute ischemic stroke,who received mechanical thrombectomy at the Kunming Yan'an Hospital and Kunming Municipal First People's Hospital,were enrolled in this study.The 315 patients who were admitted to the hospital between January 2019 and December 2023 were collected as the modeling group,and other 73 patients who were admitted to the hospital between January 2024 and September 2024 were collected as the validation group.The demographic materials and the clinical diagnosis and treatment data were collected within 24 h after admission,and the risk factors for pulmonary infection after mechanical thrombectomy were analyzed.A total of seven machine learning algorithms were employed to establish a predictive model for pulmonary infection in patients with large-vessel occlusive stroke after receiving mechanical thrombectomy.Based on the accuracy,recall,precision,F1 index,and area under the receiver operating characteristic curve(AUC),the performance of model was evaluated.Results The comprehensive performance in accuracy,recall,precision,F1 index and AUC of the gradient boosting classifier in both the modeling group and the validation group are all excellent.Conclusion The risk prediction model constructed on the basis of machine-learning algorithm technology can early identify lung infection and discover the risk factors in patients with acute ischemic stroke after receiving mechanical thrombectomy,therefore,the prevention and control measures can be promptly taken,the incidence of lung infection can be effectively decreased.

Objective:To assess the effect of low-dose esketamine on postoperative cognitive function in elderly patients undergoing non-cardiac surgery.Methods:One hundred and twenty-four patients, aged 65-80 yr, regardless of gender, with a body mass index of 18-35 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, were assigned to either esketamine group (group E, n=64) or control group (group C, n=60) using a random number table method. Group E received intravenous esketamine at a dose of 0.5 mg/kg, while group C received an equal volume of normal saline intravenously. Anesthesia was induced with intravenous fentanyl, propofol and rocuronium and maintained using combined intravenous-inhalational anesthesia in both groups. Patient-controlled analgesia was carried out postoperatively. Cognitive function tests including a simple mental state examination, auditory word learning test, tracking connection test A and B, number symbol replacement test, Boston naming test and complex graph test were performed at 1 day before surgery and 30 days after surgery, and postoperative cognitive dysfunction was determined using Z-score method. Delirium was assessed using Confusion Assessment Method from 1 to 7 days after operation. The operative hypotension, postoperative delayed emergence, nausea and vomiting, and hallucinations were recorded. The recovery time of spontaneous breathing, eye opening to verbal command and extubation time were recorded. Results:Compared with group C, the incidence of cognitive dysfunction at 30 days after surgery and intraoperative hypotension was significantly decreased ( P<0.05), and no significant change was found in the recovery time of spontaneous breathing, eye opening to verbal command, extubation time, incidence of postoperative delayed emergence, delirium, nausea and vomiting, and hallucinations in group E ( P>0.05). Conclusions:Low-dose esketamine can improve postoperative cognitive function in elderly patients undergoing non-cardiac surgery.

Objective:To investigate the effect of panax notoginseng saponins(PNS)on the expression of tumor necrosis factor-α(TNF-α)in lipopolysaccharide(LPS)-induced activated BV2 microglia through p38 mitogen-activa-ted protein kinase(p38 MAPK)pathway.Methods:BV2 microglia were divided into control group,LPS activated group and LPS+panax notoginseng saponins intervention group(LPS+PNS).The CCK-8 method was used to detect the viability of BV2 microglia and determine the optimal drug intervention concentration.Western Blot and immunofluo-rescence were used to detect the expression of p38 MAPK and TNF-α and the phosphorylation level of p38 MAPK(p-p38 MAPK)in BV2 microglia.Results:Compared with the blank control group,there was no significant difference in the cell viability of BV2 microglia,and finally 100 mg/L was selected as the drug intervention concentration.Western Blot and immunofluorescence results indicated that after LPS activation,the expression of TNF-α and the phosphoryla-tion level of p38 MAPK in BV2 microglia were significantly increased(P＜0.05).After PNS intervention,compared with LPS-activated group,the expression of TNF-α and the phosphorylation level of p38 MAPK were significantly decreased(P＜0.05).After treatment with p38 MAPK pathway inhibitor(SB203580),there was no significant differ-ence in the expression levels of p-p38 MAPK and TNF-α in PNS combined with SB203580 group(LPS+PNS+I)com-pared with LPS+PNS group(P＞0.05).In addition,the changes of p38 MAPK in each group were not statistically sig-nificant(P＞0.05).Conclusion:PNS may inhibit the expression of inflammatory factor TNF-α secreted by activated BV2 microglia through p38 MAPK pathway.

Objective:To explore the potential molecular mechanism of curcumin in the treatment of post-stroke depression through network pharmacology and molecular docking.Methods:The potential targets of curcumin were searched in GeneCards,PharmGKB and SwissTargetPrediction database.The online Mendelian Inheritance in Man data-base and GeneCards database were searched for potential targets of ischemic stroke and depression.The targets were standardized by Uniprot and then imported into Venn online platform to obtain the intersection targets among the three.Protein-protein interaction(PPI)was visualized by STRING and Cytoscape 3.7.1 software.Metascape database was used to perform Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of intersection targets.Finally,AutoDock Vina1.5.6 software was used to verify the molecular docking between curcumin and core targets.Results:A total of 1452 potential targets for ischemic stroke and depression,207 drug tar-gets,and 47 intersection targets of the three were obtained.GO functional enrichment analysis yielded 2332 items,and KEGG pathway enrichment analysis yielded 138 items.According to the degree value and enrichment fraction,the core targets such as interleukin-6(IL-6),tumor necrosis factor(TNF),serine/threonine protein kinase 1(AKT1),inter-leukin-1β(IL-1β)and the advanced glycation end products receptor(AGE-RAGE)signaling pathway,interleukin-17(IL-17)signaling pathway,TNF signaling pathway,etc.Molecular docking results showed that curcumin had a strong binding ability to core targets such as IL-6,TNF,AKT1,and 1L-1β.Conclusion:Curcumin may act on key targets such as IL-6,TNF,AKT1,IL-1β,and play an important role through the AGE-RAGE,IL-17,and TNF signaling pathways.

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.