ObjectiveTo optimize the flow cytometric sorting workflow for myeloid-derived suppressor cells （MDSCs） from murine Lewis lung carcinoma （LLC） subcutaneous tumor tissues， improving single-cell suspension preparation efficiency and sorting purity， thereby providing high-quality cells for subsequent functional studies. MethodsSubcutaneous LLC tumor tissues were harvested from C57BL/6 mice， and two enzymatic digestion protocols were first compared to evaluate their effects on single-cell suspension preparation. After staining with a viability dye （Live/Dead） and fluorochrome-conjugated anti-mouse antibodies against CD45， CD11b， and Gr-1， cells were loaded onto a Beckman CytoFLEX cell sorter. Next， conventional and optimized gating strategies were applied. In the optimized strategy， the CD45⁺ population was used as the initial gate to directly define an MDSC-enriched region， followed by selection of CD11b⁺Gr-1⁺ cells. Finally， purity was assessed by flow cytometry， viability was determined by trypan blue staining， and RT-qPCR was performed to measure the expression of MDSC signature genes， including Arg1， Nos2， IL-10， and S100A8/A9. ResultsThe optimized enzymatic digestion protocol significantly increased the yield and viability of single cells. After optimization of the gating strategy， the post-sort positivity rate （purity） was markedly improved. Arg1， Nos2， IL-10 and S100A8/A9 were highly expressed in the sorted cells， indicating that the isolated cells retained characteristic MDSC features and functional signatures. ConclusionBy optimizing both the tissue digestion protocol and the flow cytometric gating strategy， we establish an efficient workflow for isolating MDSCs from tumor tissues. This method improves the yield and purity of single-cell preparations while preserving antigen integrity， providing a reliable methodological foundation for subsequent studies on MDSC function and metabolism.

OBJECTIVE To establish an ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method for the simultaneous determination of the contents of six Aconitum alkaloids in Compound chondroitin sulfate tablets， and to perform quality evaluation of samples from different batches， thereby providing a reference for the quality control of this preparation.METHODS Chromatographic separation was performed on an Agilent SB-C 18 column using gradient elution with 0.2% formic acid （adjusted to pH 7.5 with ammonia） and acetonitrile as the mobile phases. The flow rate was 0.3 mL/min， the column temperature was 35 ℃， and the injection volume was 1 μL. Mass spectrometric analysis was performed using an AJS ESI + ion source in multiple reaction monitoring mode， with quantification by the external standard method. Method validation was performed. The established UPLC-MS/MS method was applied to determine the contents of the six Aconitum alkaloids in 64 batches of samples from three manufacturers， including three diester-type alkaloids （aconitine， hypaconitine， and mesaconitine） and three monoester-type alkaloids （benzoylaconine， benzoylhypaconine， and benzoylmesaconine）. Descriptive statistics， the Kruskal-Wallis H test， principal component analysis （PCA）， and Spearman’s rank correlation analysis were used to evaluate the content distribution of Aconitum alkaloids， differences among samples from different manufacturers， and correlations among the components.RESULTS Good linearity was observed for the six Aconitum alkaloids over their respective concentration ranges， with correlation coefficients （all r ＞0.99）. The relative standard deviations （RSDs） for precision， repeatability， and stability were all less than 5%. The limits of detection and quantification were no higher than 0.003 ng/mL and 0.01 ng/mL， respectively. The mean recoveries ranged from 96.43% to 102.30%， with RSDs ranging from 0.35% to 4.92%. Among the 64 batches of samples， the total content of monoester-type alkaloids （mean value of 0.034 9 mg/tablet） was markedly higher than that of diester-type alkaloids （mean value of 0.002 2 mg/tablet）. Benzoylmesaconine showed the highest content， with a mean value of 0.031 0 mg/tablet， whereas the other five components were present at relatively low levels and showed considerable inter-batch variation. No statistically significant differences were observed in the contents of the six components among samples from different manufacturers （ P ＞0.05）. PCA showed no obvious clustering trend， and significant correlations were observed among some components （ P ＜0.05）.CONCLUSIONS The established UPLC-MS/MS method enables the rapid and accurate simultaneous determination of the above-mentioned six Aconitum alkaloids in Compound chondroitin sulfate tablets. The quality evaluation of multiple batches of samples provides a reference for the quality control of this preparation.

Objective:To investigate the clinical characteristics and prognosis of T-lymphoblastic lymphoma (T-LBL).Methods:A retrospective case series study was conducted. Clinical data of patients diagnosed with T-LBL at the Affiliated Hospital of Jining Medical University from January 2013 to March 2023 were retrospectively analyzed, and their clinical characteristics and prognosis were statistically analyzed.Results:A total of 22 T-LBL patients were included. Among them, there were 19 males (86.4%) and 3 females (13.6%), and the median age at onset was 19.5 (15, 28) years old. Based on Ann Arbor staging, 3 cases (13.6%) were classified as stage Ⅰ-Ⅱ, while 19 cases (86.4%) were stage Ⅲ-Ⅳ; 10 cases (45.5%) presented with B symptoms, 12 cases (54.5%) without B symptoms; 16 cases (72.7%) showed elevated lactic dehydrogenase (LDH) level. At onset, 7 patients (31.8%) had mediastinal masses, 3 patients (13.6%) had central nervous system involvement, and 17 patients (77.3%) had bone marrow involvement. The overall response rate (ORR) and complete remission rate among the 22 patients were 81.82% (18/22) and 31.82% (7/22), respectively. The ORR was 84.21% (16/19) in 19 patients treated with ALL-like regimens. Among 3 patients treated with NHL-like regimens, 1 case achieved complete remission and 1 case achieved partial remission. Seven patients received allogeneic hematopoietic stem cell transplantation, with a median overall survival (OS) time of 22 months; the median OS time of patients without allogeneic hematopoietic stem cell transplantation was 14 months. The 3-year OS rates in the allogeneic hematopoietic stem cell transplantation group and group without allogeneic hematopoietic stem cell transplantation were 64.30% and 16.00%, and the difference in OS between the two groups was statistically significant ( P = 0.043). Two patients with disease progression prior to transplantation died of multidrug-resistant bacterial infections after transplantation. Conclusions:T-LBL is rare, and it is a highly aggressive tumor that predominantly occurs in adolescent males. Allogeneic hematopoietic stem cell transplantation can prolong OS, reduce relapse and improve the prognosis of patients.

Objective To detect Serum metabolites with label-free surface-enhanced Raman spectroscopy(SERS)for quickly distinguishing the metabolic profiles of breast cancer patients and healthy subjects.Methods A kind of Plasma nano-material was synthesized as a probe for SERS,which has also been used to detect Raman reporter mol-ecules to assess its detection capability.Serum samples from breast cancer patients and healthy subjects were col-lected and the proteins were precipitated with methanol and removed to collect serum metabolites.Probe-based SERS was used to analyze the serum metabolites of patients and explore the changes in the metabolic profiles of breast cancer patients.Results The SERS probe was synthesized and validated.An analytical method based on SERS probe was established,which achieved a linear range(LR)of 4 orders of magnitude and a limit of detection(LOD)up to 10 nmol/L.Raman spectra of serum metabolites from 5 breast cancer patients and 5 healthy subjects were analyzed to study differences in metabolite changes.Conclusions In this study,the molecular spectrum differences of serum metabolites in breast cancer patients were screened by probe-based SERS method,which pro-vides a technology support research on the metabolic changes caused by breast cancer so potentially provide a new method for fast breast cancer screening.

Objective:To investigate the clinical, pathological, and molecular biological characteristics of gastric hepatoid adenocarcinoma (HAS) in order to provide reference for clinical treatment.Methods:Thirty-two patients diagnosed with hepatoid adenocarcinoma after radical gastrectomy for gastric cancer at Shanxi Cancer Hospital were included from January 2019 to December 2021. Immunohistochemistry, in situ hybridization, and next-generation sequencing (NGS) methods were used to analyze immune markers and molecular characteristics in the pathological tissues from 32 patients with HAS. Cox regression analysis and Kaplan-Meier method were used to analyze the prognostic factors of overall survival and disease-free survival.Results:Among the 32 patients with HAS, 26 were male, 6 were female; aged 28-77 years, with an median age 62.0 (53.8, 67.2) years. Fifteen cases of HAS were located in the cardia, 10 cases in the antrum, and 7 cases in the body of the stomach. The maximum diameter of the mass was 3-10 cm, and mainly ulcerative in gross. The immunohistochemistry and in situ hybridization results showed that the positive rates of AFP, SALLA4, and Glypican-3 were 68.8% (22/32), 68.8% (22/32), 78.1% (25/32), respectively; Seven patients had microsatellite status of dMMR. Two cases of HER2 gene amplification and 2 cases of EB virus positivity. The NGS results showed that HAS was often accompanied by multiple gene mutations, with 23 cases having ≥ 2 gene mutations and 6 cases having ≥10 gene mutations. The TP53 gene had the highest mutation frequency; 4 cases had genetic structural variations; 28 cases had copy number variation. In addition, there were 7 cases of MSI-H and 9 cases of TMB-H. Follow-up results showed that 12 cases died, 9 cases developed metastasis, and the shortest survival time was 5 months.Conclusions:Gastric HAS is a type of tumor with high invasiveness and poor prognosis. The combined detection of AFP, SALLA4 and Glypican-3 can improve the diagnostic rate of tumors. dMMR/MSI-H and TMB-H patients in HAS are significantly higher than those in ordinary gastric cancer, and the high frequency mutation genes in HAS are often accompanied by multiple potential therapeutic targets. Immunotherapy combined with chemotherapy and targeted therapy are expected to become the treatment direction of HAS.

Parkinson's disease（PD） is the second most common neurodegenerative disease in the world， which seriously affects the lives of patients. With the acceleration of aging process， the number of patients continues to rise. Its main pathological features are aggregation of α-synuclein and degenerative death of dopaminergic neurons in the substantia nigra. However， the pathogenesis of PD is still unclear. According to reports， the brain-derived neurotrophic factor（BDNF）/tyrosine kinase receptor B（TrkB） signaling pathway is highly expressed and activated in dopaminergic neurons in the substantia nigra， which is closely related to neurophysiological processes such as neurogenesis， synaptic plasticity， neuroinflammation， and oxidative stress. It plays an important role in the occurrence and development of PD. At present， the treatment methods of Western medicine for PD are mainly based on drugs such as levodopa and dopamine agonists to alleviate motor symptoms， but with the increase of dose， the adverse reactions are significantly enhanced. Traditional Chinese medicine（TCM） has attracted people to explore its therapeutic effects on PD due to its characteristics of homology of medicine and food， economy， minor adverse reactions and multi-target action. Therefore， this paper systematically reviews the role of BNDF/TrkB pathway in the pathogenesis of PD and the mechanism of TCM formulas， extracts and monomers in the treatment of PD by regulating the BNDF/TrkB pathway according to retrieving the latest research reports at home and abroad， so as to provide a reference for the clinical application of related TCM and the development of new drugs for PD.

Objective:To understand the status and influencing factors of social isolation in elderly individuals with chronic diseases.Methods:A multi-stage sampling method was used to select elderly individuals from eight communities or villages in Hefei from July to September 2022. The study employed a general information survey, the Lubben Social Network Scale (LSNS-6) , the Geriatric Depression Scale (GDS-15) , the Social Support Rating Scale (SSRS) , and the Personal Social Capital Scale 16 (PSCS-16) . Binary Logistic regression analysis was used to explore the factors influencing social isolation in elderly individuals with chronic diseases.Results:A total of 1 133 elderly individuals were surveyed, among which 538 had chronic diseases. Among the 538 elderly individuals with chronic diseases, 209 were socially isolated, resulting in a social isolation rate of 38.8%. Binary Logistic regression analysis revealed that living area, fear of falling, depression, social capital, and social support were significant factors influencing social isolation ( P<0.05) . Conclusions:The social isolation rate among elderly individuals with chronic diseases is high. Special attention should be given to elderly individuals living in rural areas, those who fear falling, and those experiencing depression. Additionally, improving social capital and social support can help alleviate social isolation in these elderly individuals.

Objective:To explore the potential categories and influencing factors of attitudes to aging among the elderly in nursing homes.Methods:Convenience sampling was used to select elderly from 15 nursing homes in Anhui Province from March to July 2024 for the study. The survey was conducted using the General Information Questionnaire, Attitudes to Ageing Questionnaire, Belongingness Scale for Older Adults in Nursing Facilities, and the Lubben Social Network Scale. Mplus 8.3 was used to conduct a latent profile analysis, and multivariate Logistic regression was employed to explore the factors influencing the latent categories of attitudes toward aging among the elderly in nursing facilities.Results:A total of 370 questionnaires were distributed, and 366 valid questionnaires were recovered, with an effective recovery rate of 98.9%. The attitudes to aging of 366 elderly of nursing facilities were expressed in three potential categories, namely, negative-perceived type (48.6%), moderate-low psychosocial loss type (35.0%), and positive and optimistic-comprehensive type (16.4%). Multicategorical Logistic regression analysis showed that the number of children, location of the nursing home, educational level, self-rating health, belongingness to the nursing home, and social network were the influencing factors of the potential categories of attitudes to aging among the elderly in the nursing home, and the difference was statistically significant ( P<0.05) . Conclusions:There is group heterogeneity in the attitudes to aging among the elderly in nursing homes. Healthcare professionals should focus on negative-perceived elderly and provide targeted interventions based on the potential categories of factors influencing their attitudes to aging.

Non-alcoholic fatty liver disease(NAFLD)is a chronic liver disease caused by excessive lipid accumulation in the liver.Its incidence rate is increasing year by year and has become an increasingly serious public healthy problem.The pathogenesis of NAFLD is complex and has not been fully clarified at present.It is mainly related to multiple factors such as genetics,metabolism,intestinal flora and immune response.In order to explore the medication rules and mechanism of action of traditional Chinese medicine(TCM)in the treatment of NAFLD,and to provide references for the treatment of NAFLD with TCM and the research and development of new drugs,this article summarizes the TCM pathogenesis of NAFLD(such as"phlegm and blood stasis interlacing","liver depression and spleen deficiency",etc.)and modern etiology and pathogenesis(such as insulin resistance,lipid disorder,mitochondrial dysfunction,oxidative stress,etc.).The clinical research and experimental data at home and abroad in recent years were integrated to analyze the pathological process of NAFLD intervention by TCM through multiple targets,including improving insulin resistance and lipid metabolism disorders,inhibiting oxidative stress and mitochondrial dysfunction,etc.TCM has shown unique advantages in the prevention and treatment of NAFLD.However,the depth of its mechanism analysis and the level of clinical research still need to be improved.In the future,it is necessary to deepen the mechanism research by combining multi-omics technology to accelerate the modernization development of TCM.

Immunogenic cell death(ICD)is a form of cell death that can activate the immune system,especially in the treatment of cancer.ICD can enhance the recognition of tumors by the immune system and the release of damage associated molecular patterns(DAMPs),to achieve tumor cell death.Oncolytic viruses(OVs)can selectively infect and kill tumor cells without damaging normal cells.OVs are type Ⅱ ICD inducers that induce ICD in tumor cells by targeting the endoplasmic reticulum.Here,we review the characteristics of ICD and the mechanism of ICD induction by OVs.We also review the latest clinical progress involving ICD and discuss future treatment strategies for tumors.