In maintaining normal function and activation processes, glycolysis, lipid metabolism, and amino acid metabolism play key roles in the energy demand of platelets. In the resting state, platelets primarily rely on glycolysis and aerobic oxidation to generate energy. Upon activation, platelets preferentially utilize glycolysis, as it can more rapidly provide the required ATP. In addition to glycolysis, platelets can also utilize glycogen and fatty acids as additional energy sources. The ATP provided by fatty acid oxidation is crucial for platelet activation. Additionally, during platelet storage, distinctive changes in energy metabolism occur. In the early stages of storage, platelets primarily rely on glycolysis and the pentose phosphate pathway (PPP) to generate energy. In the mid-storage phase, there is an increase in tricarboxylic acid cycle (TCA) metabolism. In the later stages of storage, cellular metabolism gradually declines. The regulation and flexibility of these metabolic pathways play a critical role in the survival and function of platelets in different states.

Traditional Chinese medicine （TCM） has unique advantages in alleviating the symptoms of diabetic kidney disease （DKD） and slowing its progression. However， traditional clinical trials often use the occurrence of end-stage renal disease as the end point， requiring long-term follow-up， which increases trial complexity and costs， thereby limiting the feasibility of TCM clinical studies. This paper suggested that in clinical trials of TCM for DKD， both the estimated glomerular filtration rate （eGFR） change rate （≥30%） and eGFR slope can serve as potential surrogate outcome measures. If the intervention course is short （＜1 year）， the eGFR change rate （≥30%） is recommended as a surrogate outcome measure， whereas in long-term interventional studies （≥1 year）， the eGFR slope may be more appropriate. Furthermore， based on biochemical indicators such as eGFR slope and urinary albumin-to-creatinine ratio （UACR） change rate， integrating TCM symptom evaluation， TCM syndrome evaluation， and quality of life scales can help develop internationally recognized patient-reported outcome measures （PROMs） for TCM clinical trials， which will be a key step in enhancing the evaluation system for the effectiveness of TCM in treating DKD.

Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO BA. 04/ABO O.01.01 and ABO B3.05/ABO O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.

Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO BA. 04/ABO O.01.01 and ABO B3.05/ABO O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.

A new method for simultaneous determination of 23 kinds of per-and polyfluoroalkyl substances(PFASs)(13 kinds of perfluoro carboxylic acids,4 kinds of perfluoro sulfonic acids,and 6 kinds of new substitutes)in plant leaf tissue by ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)using automatic online solid phase extraction(SPE)to remove the matrix interference components in plant crude extracts was developed.The plant leaf samples were extracted twice with 1%formic acid-methanol solution,then evaporated to dry,redissolved with 70%methanol solution,and directly injected for analysis.After 23 kinds of target PFASs were purified automatically by online SPE with a WAX column,the six-way valve was switched to rinse PFASs onto an alkaline mobile phase system-compatible C18 analytical column.Then,the 23 kinds of target PFASs were separated within 16 min by gradient elution using a binary mobile phase system of methanol/water(Containing 0.4%ammonium hydroxide).Tandem mass spectrometry was performed in multiple reaction monitoring(MRM)mode for online detection of various PFASs,and quantification was carried out by internal standard method.The results of the method validation showed that satisfactory average recoveries of 23 kinds of PFASs in plant leaf samples(64.2%-125.5%),precision(relative standard deviations(RSDs)of 0.7%-12.8%),linearity(R2＞0.990),and sensitivity(the detection limits(S/N=3)were in the range of 0.02-0.50 μg/kg)were achieved.Finally,this method was used to detect PFASs in the marine green tide algae(Enteromorpha prolifera)and several tree leaves,and a total of 6 kinds of PFASs were detected,in which PFBA was the main contaminant.Compared with the reported offline SPE methods,the proposed online SPE technique significantly simplified the sample pretreatment process and provided an automatic,simple,and environment-friendly method for the routine monitoring of legacy and emerging PFASs in plant tissues.

In order to solve the problem of resistance of Pseudomonas aeruginosa to multiple antibiotics, it is an effective way to find inhibitors of P. aeruginosa efflux pump. In this study, 15 new ornithine peptidomimetic derivatives were designed and synthesized by changing the side chain structure of natural amino acids with PAβN, a dipeptide efflux pump inhibitor, and their synergic activity with aztreonam, a monocyclic β-lactam antibiotic, against P. aeruginosa was evaluated. Among them, the representative compound 12b not only enhanced the antibacterial activity of β-lactam antibiotics aztreonam, ceftazidime and meropenem, but also significantly enhanced the antibacterial action of macrolide antibiotics clarithromycin, showing a broad-spectrum synergic sensitization effect. In addition, compound 12b also has a good safety. Preliminary mechanisms suggest that 12b works by directly targeting the efflux transporter MexB. These results provide a new lead compound for the development of a new class of efflux pump inhibitors against P. aeruginosa.

Objective:By activating gamma-aminobutyric acid(GABA)neurons in zona incerta(ZI),we observed the alleviation of motor ability impairment in mice treated with lipopolysaccharide(LPS),and investigate the function of GABA neurons in the process of motor function impairment in mice with neuroinflammation.Methods:The central nervous system infection model of C57BU6 mice was eatablished by intraperitoneal injection of LPS.The motor function were detected by open field test and running wheel test in mice.The expression of c-Fos in ZI neurons and ionized calci-um binding adaptor molecule 1(Iba-1)in glial cells were detected by immunofluorescence staining.rAAV-DIO-hM3Dq-mCherry was bilaterally injected into ZI of adult male VGAT-Cre mice through the stereotaxic technique.After three weeks with the virus sufficiently expressing in the neurons,the neuroinflammation model was made by intrabitoneal injection of LPS,and GABAergic neurons were activated by intrabitoneal injection of N-oxyclozapine(CNO)30 min before behavioral tests.The motor ability in mice were detected by open field test and running wheel test.Results:The results of open field experiment and wheel experiment showed that the activity of mice in LPS group was significantly decreased(P＜0.05)compared with the control group.The expression of ZI c-Fos positive protein in the brain of mice was significantly down-regulated with Iba-1 positive protein up-regulating in the ZI.The chemogenetic results showed that the activity of LPS mice was relieved with ZI GABAergic neurons being activated(P＜0.05).Conclusion:The activity of ZI GABAergic neurons was significantly decreased and the motor ability was impaired in mice with neuroin-flammation in the brain.Chemogenetic activation of ZI GABAergic neurons significantly improved the motor ability of inflammatory mice.

Objective:Proximal femur tumor resection often leads to hip joint instability and functional loss.Various methods have been clinically applied to repair hip joint soft tissue function,but deficiencies remain.This study aims to evaluate the advantages and disadvantages of the ligament advanced reinforcement system(LARS)tumor tube in assisting soft tissue function reconstruction in patients undergoing tumor type artificial hip replacement surgery. Methods:This study included 85 patients(41 males,44 females)with proximal femoral tumors treated at the Xiangya Bone Tumor Treatment Center from January 2012 to January 2022,aged 10 to 79(38.5±18.2)years.Among them,13 cases had benign aggressive tumors,45 had primary malignant bone tumors,and 27 had bone metastases.Clinical data,imaging data,and intraoperative photos were collected.Patients were followed up and postoperative functional evaluations were conducted using the Musculoskeletal Tumor Society(MSTS)scoring system and Harris hip joint scoring system to assess limb function and hip joint function. Results:Preoperative pathological fractures were present in 37 cases(43.5％),with a lesion length of(9.4±2.9)cm.Among non-metastatic tumor patients,7 experienced postoperative recurrence,including 6 cases of osteosarcoma and 1 case of fibrosarcoma.Pulmonary metastases occurred in 9 osteosarcoma patients.Five patients required reoperation due to postoperative complications,including 3 cases of deep vein thrombosis,1 case of giant cell granuloma,and 1 case of prosthesis infection.Postoperatively,5 patients exhibited Trendelenburg gait,and 6 had leg length discrepancies.The postoperative MSTS score was 26.7±1.4,and the Harris score was 89.6±5.3. Conclusion:The LARS tumor tube can effectively assist in reconstructing the soft tissue function of the hip joint and greatly reduce postoperative complications,making it an effective technical improvement in joint function reconstruction in tumor type artificial hip replacement surgery.

Objective To explore the efficacy of ransapical-transcatheter aortic valve replacement(TA-TAVR)in patients with severe aortic regurgitation(AR)with different ejection fraction.Methods The clinical data of patients with severe AR undergoing TA-TAVR from November 2017 to January 2023 were retrospective reviewed,severe AR patients with left ventricular ejection fraction(LVEF)≥50％were included in the ejection fraction preserved group and patients with severe AR with LVEF＜50％were included in the reduced ejection fraction group.Clinical data of the patients in group 2 were analyzed.Results A total of 60 patients were included,including 24 in ejection fraction preserved group and 36 in reduced ejection fraction group.All group had significantly improved aortic regurgitation volume 1 day after surgery compared with preoperative volume.The left ventricular remodeling(LVR)indexes[left ventricular end-diastolic diameter(LVEDd),left ventricular end-systolic diameter(LVESD),left ventricular posterior wall thickness(LVPW),LVEF]improved significantly at 1 day after surgery,and remained relatively stable thereafter.The improvement in LVR in reduced ejection fraction group patients was only 3 months after surgery.Two groups of patients had a decrease in amino-terminal probrain natriuretic peptide(NT-proBNP)levels three months after surgery compared to before.Patients in ejection fraction preserved group remained stable thereafter,while patients in reduced ejection fraction group had an increase one year after surgery compared to before.The one-year heart failure rehospitalization rate in reduced ejection fraction group was higher than that in ejection fraction preserved group,and the mean scores of EQ-5D-3L and EQ-VAS were lower than those of patients in ejection fraction preserved group.Comparison of the incidence of atrial fibrillation,third degree atrioventricular block,and permanent pacemaker implantation rate,stroke incidence,leaflet thrombosis incidence,paravalvular leakage incidence,and survival rate,the usage of some drugs at 1 year in both group,there was no statistically significant difference(all P＞0.05).Conclusions TA-TAVR can effectively relieve aortic regurgitation in severe AR patients with different ejection fraction and promote left ventricular reverse remodeling.Intervention in patients with severe AR with preserved ejection fraction should be performed as early as possible.

OBJECTIVE: Physical exercise, a common non-drug intervention, is an important strategy in cancer treatment, including hepatocellular carcinoma (HCC). However, the mechanism remains largely unknown. Due to the importance of hypoxia and cancer stemness in the development of HCC, the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness. METHODS: A physical exercise intervention of swimming (30 min/d, 5 d/week, for 4 weeks) was administered to BALB/c nude mice bearing subcutaneous human HCC tumor. The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring, hematoxylin and eosin (HE) staining, and immunohistochemistry (IHC) detection of proliferating cell nuclear antigen (PCNA) and Ki67. The expression of stemness transcription factors, including Nanog homeobox (NANOG), octamer-binding transcription factor 4 (OCT-4), v-Myc avian myelocytomatosis viral oncogene homolog (C-MYC) and hypoxia-inducible factor-1α (HIF-1α), was detected using real-time reverse transcription polymerase chain reaction. A hypoxia probe was used to explore the intratumoral hypoxia status. Western blot was used to detect the expression of HIF-1α and proteins related to protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway. The IHC analysis of platelet endothelial cell adhesion molecule-1 (CD31), and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion. SMMC-7721 cells were treated with nude mice serum. The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors. The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1α. Further, the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected. RESULTS: Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor. HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention. Swimming potently alleviated the intratumoral hypoxia, attenuated the cancer stemness, and inhibited the Akt/GSK-3β/β-catenin signaling pathway. Additionally, the desmin⁺/CD31⁺ ratio, rather than the number of CD31⁺ vessels, was significantly increased in swimming-treated mice. In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere, as well as the mRNA expression level of stemness transcription factors. Consistent with the in vivo results, HIF-1α and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group. CONCLUSION Swimming alleviated hypoxia and attenuated cancer stemness in HCC, through suppression of the Akt/GSK-3β/β-catenin signaling pathway. The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor. Please cite this article as: Xiao CL, Zhong ZP, Lü C, Guo BJ, Chen JJ, Zhao T, Yin ZF, Li B. Physical exercise suppresses hepatocellular carcinoma progression by alleviating hypoxia and attenuating cancer stemness through the Akt/GSK-3β/β-catenin pathway. J Integr Med. 2023; 21(2): 184-193.
Humans ; Animals ; Mice ; Carcinoma, Hepatocellular/drug therapy* ; Proto-Oncogene Proteins c-akt/metabolism* ; Proliferating Cell Nuclear Antigen/therapeutic use* ; Mice, Nude ; Glycogen Synthase Kinase 3 beta/genetics* ; beta Catenin/therapeutic use* ; Liver Neoplasms/drug therapy* ; Desmin/therapeutic use* ; Ki-67 Antigen ; Cell Line, Tumor ; Hypoxia ; RNA, Messenger/therapeutic use* ; Cell Proliferation