Objective To investigate the effects of BMSCs treatment on neuropathic pain in rats with SCI and explore the underlying mechanism.Methods The rats were randomly divided into the following groups(n=15 per group):sham-operated(sham)group,spinal cord injury model(SCI)group,SCI+BMSCs group,and SCI+BMSCs+LY294002 group.An SCI model was established using Sprague-Dawley rats,followed by intraspinal administration of BMSCs and the PI3K inhibitor LY294002 to the injured spinal cord of SCI rats.The BBB score,pMWT,and pTWL values under thermal stimulation were measured.Hematoxylin-eosin staining was used to observe pathological changes in the injured spinal cord of rats.The effects of BMSCs transplantation on SCI rats were explored through hematoxylin-eosin staining,immunofluorescence staining,ELISA,and Western Blot experiments.RSCN were induced using LPS and co-cultured with BMSCs and their exosomes.The effects of BMSCs and their exosomes on RSCN were investigated through Annexin V-FITC/PI kit,ELISA,and Western Blot assays.Results The SCI model was considered successfully established when the following criteria were met on post-operative day 5:BBB locomotor score≤5,accompanied by a BBB score<10 on day 20,along with histopathological evidence of spinal cord tissue loosening,extensive vacuolation,and neuronal atrophy observed via HE staining.Compared with the sham group,the SCI group exhibited significantly lower BBB scores,pMWT,and pTWL values(P<0.001).Concurrently,increased immunofluorescence intensity of IBA1,elevated levels of pro-inflammatory cytokines,and pain-related factors were detected in the spinal cord(P<0.001).Furthermore,activation of the PI3K/AKT signaling pathway was significantly suppressed.BMSCs transplantation protected SCI rats by activating the PI3K/AKT pathway(P<0.001).BMSC-mediated spinal cord repair was attenuated by LY294002 administration.LPS-induced RSCNs showed increased apoptosis and pro-inflammatory cytokine release(P<0.001).Co-culture with MSCs or BMSCs-derived exosomes activated the PI3K/AKT signaling pathway,thereby reducing LPS-induced apoptosis and proinflammatory cytokine production(P<0.05).Conclusion BMSCs activate the PI3K/AKT signaling pathway in neurons through exosomes,suppressing the levels of TNF-α,SP,NE,and 5-HT,and promoting functional recovery in SCI rats.

OBJECTIVE To explore the willingness to pay （WTP） of Urumqi residents for community pharmacies’ medication guidance services and its influencing factors， so as to provide data support for the optimization of community pharmacy services and the establishment of a fee structure for medication guidance services. METHODS A stratified quota sampling method was employed to select 14 communities in Urumqi City. From April to June 2025， a combined offline and online questionnaire survey was conducted among adult residents in these communities. The contingent valuation method was used to construct three hypothetical scenarios （namely， basic， enhanced and extended services） of medication counselling in community pharmacies to assess residents’ WTP for these services. Binary Logistic regression was employed to analyze the influencing factors of WTP. RESULTS A total of 576 valid questionnaires were obtained. Under the scenarios of basic， enhanced and extended services， 38.54%， 49.65% and 67.19% of the respondents expressed WTP for the services， respectively. Occupational type， type of basic medical insurance， annual income， perception of pharmacists’ profession， and acceptance level of the service were identified as major influencing factors for WTP （P＜0.05）. CONCLUSIONS The willingness of residents in Urumqi to pay for medication counseling services provided by pharmacists in community pharmacies significantly increases with the enrichment of service content. It is recommended to incorporate basic medication counselling services provided by pharmacists in community pharmacies into medical insurance payment， while value-added services should be partially or fully self-paid by residents. Additionally， efforts should be made to strengthen the promotion of the professional and service value of licensed pharmacists， so as to facilitate the high-quality development of pharmaceutical care.

Objective To investigate the expression and clinical significance of B7-H5 in patients with chro-nic hepatitis B and hepatitis B virus(HBV)-related hepatocellular carcinoma.Methods Enzyme-linked immu-nosorbent assay(ELISA)was used to detect the serum levels of B7-H5 in 104 patients with chronic hepatitis B,28 patients with HBV-related hepatocellular carcinoma and 35 healthy controls.And statistical methods were used to analyze the difference,correlation and diagnostic value of the results.Results The serum levels of B7-H5 in patients with HBV-related hepatocellular carcinoma,patients with chronic hepatitis B and healthy subjects were statistically different(P＜0.05),and the level from high to low was HBV-related liver cancer patients,chronic hepatitis B patients,healthy subjects.The serum level of B7-H5 in chronic hepatitis B pa-tients with negative HBV-DNA was significantly higher than that in healthy subjects(P＜0.01).The level of B7-H5 in patients with chronic hepatitis B was positively correlated with HBV-DNA load(P＜0.05).And the level of B7-H5 in patients with HBV-related hepatocellular carcinoma was positively correlated with HBsAg,HBeAg and AFP levels(P＜0.05).In addition,B7-H5 had diagnostic value for chronic hepatitis B and HBV-related hepatocellular carcinoma(P＜0.01),and the diagnostic value for HBV-related hepatocellular carcino-ma was higher than that for chronic hepatitis B.The level of B7-H5 in patients with HBV-related hepatocellu-lar carcinoma after surgery was significantly lower than that before surgery(P＜0.05).Conclusion Serum levels of B7-H5 is related to the progression of CHB and HBV-related hepatocellular carcinoma,and can be considered as an effective detection index for the auxiliary diagnosis and the judgment of postoperative condi-tion of CHB and HBV-related hepatocellular carcinoma.

Objective:To study the effect of anthocyanins(C3G)on cuproptosis in chronic epileptic rats.Methods:Chronic epileptic rat model was induced by pentatetrazol(PTZ),and 90 SD rats were randomly divided into control group,PTZ group,elesclomol(ELC)group,tetrathiomolybdate(TTM)group,C3G group and ELC+C3G group.The grade,latency and frequency of seizures were recorded in each group.electroencephalogram(EEG)was used to detect abnormal electrical discharge in the brain.The action potential of hippocampal neurons was measured by patch-clamp technique.The contents of glutathione(GSH)and cuprous ions(Cu+)in hippocampus were determined by kit.Neuron damage in hippocampus was evaluated by Nissl staining.The expression of ferredoxin1(FDX1)and lipoic acid synthase(LIAS)in hippocampus was analyzed by immunohistochemistry and Western blot.Results:Compared with the control group,the rats in the PTZ group exhibited epileptic-like seizures,suggesting that the modeling was successful.Com-pared with other epileptic groups,the ELC group showed increased seizure grade,more abnormal discharges,shortened latency period,enhanced neuronal excitability,decreased Nissl particles,elevated Cu+levels,decreased GSH levels,and increased expressions of FDX1 and LIAS.The reverse was observed in C3G group(P＜0.05).Neuron damage in ELC+C3G group was less severe than that in ELC group,but more than that in PTZ group(P＜0.05).Neuron dam-age in TTM group was less severe than that in PTZ group,but more severe than that in C3G group(P＜0.05).Conclusion:cuproptosis exists in hippocampus of rats with chronic epilepsy,and the C3G can significantly inhibit cu-proptosis and alleviate the occurrence and development of chronic epilepsy.

Ovulatory dysfunction (OD) is one of the main causes of infertility in women of childbearing age, which not only affects their reproductive ability, but also physical and mental health. Traditional treatment strategies have limited efficacies, and the emergence of biomedicines provides a promising alternative solution via the strategies of combining engineered design with modern advanced technology. This review explores the pathophysiological characteristics and related induction mechanisms of OD, and evaluates the current cutting-edge advances in its treatments. It emphasizes the potentials of biomedicines strategies such as hydrogels, nanoparticles and extracellular vesicles in improving therapeutic precision and efficacy. By mimicking natural physiological processes, and achieving controlled drug release, these advanced drug carriers are expected to address the challenges in ovarian microenvironment reprogramming, tissue repair, and metabolic and immune regulation. Despite the promising progress, there are still challenges in terms of biomedical complexity, differences between animal models and human physiology, and the demand for intelligent drug carriers in the therapy of OD. Future researches are mainly dedicated to developing precise personalized biomedicines in OD therapy through interdisciplinary collaboration, promoting the development of reproductive regenerative medicine.

OBJECTIVES: To investigate the active components and possible mechanisms of n-butanol fraction of Periploca forrestii Schltr. ethanol extract for treating Alzheimer's disease (AD). METHODS: The active components of n-butanol fraction of Periploca forrestii Schltr. ethanol extract were analyzed using UPLC-QE-MS technique. In a SD rat model of AD induced by treatment with AlCl₃ and D-gal, the therapeutic effects of low, moderate and high doses of the n-butanol fraction, saline, and donepezil hydrochloride were evaluated using ELISA, HE and Nissl staining, immunohistochemistry and Western blotting. The therapeutic mechanisms of the n-butanol fraction were explored using network pharmacology and molecular docking. RESULTS: Seventeen active components were identified from the n-butanol fraction of Periploca forrestii Schltr. ethanol extract, including phenylpropanoids, flavonoids, anthraquinones, triterpenoids, steroids, and volatile oils. In the rat models of AD, treatment with the n-butanol fraction significantly lowed AChE content in the hippocampus, increased the contents of ACh, SOD, CAT, and GSH-Px, enhanced the expressions of neuronal apoptotic factors Bcl-2, PI3K, Akt, p-PI3K, and p-Akt, and reduced the expressions of Bax and caspase-3 proteins. The treatment also dose-dependently up-regulated hippocampal expressions of Nrf-2, HO-1 and BDNF and down-regulated Keap-1, Aβ and Tau expressions. Bioinformatics analysis identified 14 key intersected targets (including TNF, AKT1 and ESR1) between the n-butanol fraction and AD. CONCLUSIONS The therapeutic effect of n-butanol fraction of Periploca forrestii Schltr. ethanol extract in AD mice is mediated by its multiple active components that regulate multiple targets and pathways.
Animals ; Rats, Sprague-Dawley ; Rats ; 1-Butanol/chemistry* ; Plant Extracts/pharmacology* ; Periploca/chemistry* ; Ethanol/chemistry* ; Alzheimer Disease/drug therapy* ; Male ; Molecular Docking Simulation ; Apoptosis/drug effects*

Aim To investigate the protective and reparative effect of empagliflozin on oxidized-low density lipo-protein(ox-LDL)-induced injury in human umbilical vein endothelial cells(HUVEC)and its mechanism of action.Methods Primary HUVEC were cultured in vitro.ox-LDL was used to induce HUVEC injury model,and the cell sur-vival rate was measured by CCK-8 assay.EDU method was used to detect cell proliferation.Western blot was used to detect the protein levels of Ki-67,Bcl-2,cleaved Caspase-3,Bax,endothelial nitric oxide synthase(eNOS),intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule 1(VCAM-1)and epidermal growth factor receptor(EGFR)in HUVEC.RT-qPCR was used to detect the mRNA expression levels of interleukin-6(IL-6)and tumor necro-sis factor-α(TNF-α)in HUVEC.Using Swiss targets,GeneCards databases,gene ontology(GO)and Kyoto encyclope-dia of genes and genomes(KEGG),protein-protein interaction(PPI)network analysis,and ClickDocking(https://mcule.com/apps/1-click-docking/)to predict the target of empagliflozin.Results CCK-8 results showed that 0.025 μmol/L empagliflozin significantly alleviated ox-LDL-induced HUVEC injury(P＜0.01).EDU results showed that ox-LDL treatment for 24 h significantly inhibited the proliferation of HUVEC(P＜0.01),while empagliflozin treatment sig-nificantly alleviated the inhibition of cell proliferation(P＜0.01).The results of Western blot showed ox-LDL treatment significantly decreased the protein expression levels of Ki-67,Bcl-2,and eNOS,and increased the protein expression levels ofcleaved Caspase-3,Bax,ICAM-1,and VCAM-1 in HUVEC(all P＜0.05).However,empagliflozin treatment reversed these changes(all P＜0.05).RT-qPCR results showed that ox-LDL treatment increased the mRNA expression levels of IL-6 and TNF-α in HUVEC(P＜0.01),while empagliflozin treatment decreased their expression levels(P＜0.05).However,after adding EGFR agonist NSC 228155,the protective effect of empagliflozin against ox-LDL-mediated HUVEC injury was significantly reversed(P＜0.05).Conclusion Empagliflozin can significantly reduce ox-LDL-in-duced HUVEC injury,which may be related to EGFR signaling pathway.

Objective:To investigate protective effect and mechanism of Tim-3 on sepsis-induced acute lung injury(ALI)by pro-moting mitophagy of alveolar macrophages and inhibiting activation of NLRP3 inflammasome.Methods:LPS-stimulated mouse alveo-lar macrophage(MH-S)model and sepsis-induced ALI mouse model were constructed.Tim-3 siRNA interference technique was used to knock down Tim-3 expression in MH-S cells,and anti-Tim-3 antibody mice were injected intraperitoneally to block Tim-3 function.Western blot was used to detect protein expressions of NLRP3,ASC,cleaved-caspase-1 and mitophagy-related proteins(LC3B,P62,PINK1 and Parkin)in MH-S cells and lung tissue of mice with sepsis-induced ALI.Laser confocal fluorescence staining was used to measure ROS level and mitochondrial membrane potential of MH-S cells.Pathological examination of lung tissue was performed in mice with sepsis-induced ALI in each group,and degree of lung tissue injury was evaluated by Smith scoring system.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected from mice with ALI induced by sepsis in each group.BCA protein quantification method was used to determine protein concentration in BALF.MPO activity in lung tissue was detected by colorimetry.MDA content in lung tissue was detected by TBA method.LC3B protein expression in lung tissue was detected by immunohistochemistry.Results:In mouse alveolar macrophages,Tim-3 knockdown could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins,increase ROS release,inhibit PINK1/Parkin pathway activation and LC3B protein expression,and reduce mitochondrial membrane potential.In mice with sepsis-induced ALI,Tim-3 functional blockade could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins in lung tissue,aggravate lung pathological injury and pulmonary edema,increase MPO activity and MDA content in lung tissue,and reduce positive rate of LC3B protein.Conclusion:Tim-3 plays a protective role in sepsis-induced ALI by promoting mitophagy in alveolar macrophages and inhibiting NLRP3 inflammasome activation via PINK1/Parkin.

Aim To investigate the protective and reparative effect of empagliflozin on oxidized-low density lipo-protein(ox-LDL)-induced injury in human umbilical vein endothelial cells(HUVEC)and its mechanism of action.Methods Primary HUVEC were cultured in vitro.ox-LDL was used to induce HUVEC injury model,and the cell sur-vival rate was measured by CCK-8 assay.EDU method was used to detect cell proliferation.Western blot was used to detect the protein levels of Ki-67,Bcl-2,cleaved Caspase-3,Bax,endothelial nitric oxide synthase(eNOS),intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule 1(VCAM-1)and epidermal growth factor receptor(EGFR)in HUVEC.RT-qPCR was used to detect the mRNA expression levels of interleukin-6(IL-6)and tumor necro-sis factor-α(TNF-α)in HUVEC.Using Swiss targets,GeneCards databases,gene ontology(GO)and Kyoto encyclope-dia of genes and genomes(KEGG),protein-protein interaction(PPI)network analysis,and ClickDocking(https://mcule.com/apps/1-click-docking/)to predict the target of empagliflozin.Results CCK-8 results showed that 0.025 μmol/L empagliflozin significantly alleviated ox-LDL-induced HUVEC injury(P＜0.01).EDU results showed that ox-LDL treatment for 24 h significantly inhibited the proliferation of HUVEC(P＜0.01),while empagliflozin treatment sig-nificantly alleviated the inhibition of cell proliferation(P＜0.01).The results of Western blot showed ox-LDL treatment significantly decreased the protein expression levels of Ki-67,Bcl-2,and eNOS,and increased the protein expression levels ofcleaved Caspase-3,Bax,ICAM-1,and VCAM-1 in HUVEC(all P＜0.05).However,empagliflozin treatment reversed these changes(all P＜0.05).RT-qPCR results showed that ox-LDL treatment increased the mRNA expression levels of IL-6 and TNF-α in HUVEC(P＜0.01),while empagliflozin treatment decreased their expression levels(P＜0.05).However,after adding EGFR agonist NSC 228155,the protective effect of empagliflozin against ox-LDL-mediated HUVEC injury was significantly reversed(P＜0.05).Conclusion Empagliflozin can significantly reduce ox-LDL-in-duced HUVEC injury,which may be related to EGFR signaling pathway.

OBJECTIVE: To report the blood group antigen and antibody specificity identification methods for a patient with high-frequency antibodies, and the process of finding and providing compatible blood for the patient. METHODS: A patient sent from the Blood Transfusion Department of Shanxi Provincial People's Hospital to Blood Transfusion Technology Research Laboratory of Taiyuan Blood Center in November 2022 was selected for the study. Classical serological methods were used to determine the patient's blood type, screen for unexpected antibodies, identify antibodies, and perform crossmatching. High-frequency antibody identification was carried out using red blood cells treated with various enzymes. Blood group genotyping was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and Sanger sequencing. Multiple strategies were employed to address the patient's blood source problem. The study was approved by the Medical Ethics Committee of Taiyuan Blood Center [Ethics No. 2024 Ethics Review No.(2)]. RESULTS: The patient's blood type was B, RhD positive. Initial screening of the patient's serum with multiple screening cells and antibody identification cells in saline medium was negative, but positive in antiglobulin medium. The patient's serum showed varying reaction intensities with red blood cells treated with different enzymes. MALDI-TOF mass spectrometry and Sanger sequencing revealed a homozygous nonsense variant c.376C>T (p.Gln126Ter) in the ABCG2 gene, resulting in the Jr(a-) phenotype. During family donor selection, the patient's son was found to have a heterozygous variant c.376C>T (p.Gln126Ter), and another heterozygous variant c.421C>A (p.Gln141Lys), which predicted a Jr(a+w) phenotype. Crossmatch tests confirmed the compatibility of blood from the patient's son, which was used to address the urgent blood requirement. Later, rare blood from a Jr(a-) donor from the Guangzhou Blood Center was used for the patient's ongoing treatment, saving the patient's life. CONCLUSION Combining classic serological testing with blood group gene typing techniques successfully identified the rare Jr(a-) blood type and high-frequency anti-Jra antibodies. Enzyme-treated red blood cell identification methods confirmed the presence of anti-Jra antibodies. By searching within the family and seeking help from other blood centers, compatible blood was found. This approach may provide insights for resolving similar complex blood matching problems in the future.
Humans ; Blood Grouping and Crossmatching/methods* ; Blood Group Antigens/immunology* ; Blood Transfusion ; Male ; Isoantibodies/blood* ; Female ; Genotype