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.

ObjectiveTo explore the effect of respiratory training based on core stabilization training on lumbar disc herniation. MethodsFrom January, 2023 to October, 2024, 96 patients with lumbar disc herniation admitted to the First Affiliated Hospital of Bengbu Medical University were divided into control group (n = 32), core group (n = 32) and respiratory group (n = 32). All the groups underwent conventional rehabilitation therapy, with core stabilization training in the core group and respiratory training combined with core stabilization training in the respiratory group, additionally, for four weeks. Before and after training, the scores of Visual Analogue Scale, Japanese Orthopaedic Association (JOA) and Oswestry Dysfunction Index (ODI) were compared, the average electromyographic value (AEMG) and root mean square (RMS) value of the multifidus and transversus abdominis were detected by surface electromyography (sEMG); and the thickness of the multifidus and transversus abdominis were measured by musculoskeletal ultrasonography bilaterally. ResultsThe intra-group effect (F > 597.796, P < 0.001), inter-group effect (F > 16.535, P < 0.001) and interaction effect (F > 49.622, P < 0.001) were significant in the scores of VAS, JOA and ODI; which were better in the respiratory group than in the control group and the core group (P < 0.05), and were better in the core group than in the control group (P < 0.001). The intra-group effect (F > 7971.631, P < 0.001), inter-group effect (F > 177.760, P < 0.001) and interaction effect (F > 478.771, P < 0.001) were significant in the thickness of the transversus abdominis and multifidus; which were better in the respiratory group than in the control group and the core group (P < 0.001), and were better in the core group than in the control group (P < 0.001). The intra-group effect (F > 144303.007, P < 0.001), inter-group effect (F > 1495.458, P < 0.001) and interaction effect (F > 3121.361, P < 0.001) were significant in the RMS of the multifidus and transversus abdominis; which were better in the respiratory group than in the control group and the core group (P < 0.001), and were better in the core group than in the control group (P < 0.001). The intra-group effect (F > 1890.532, P < 0.001), inter-group effect (F > 607.132, P < 0.001) and interaction effect (F > 824.923, P < 0.001) were significant in the AEMG of the multifidus and transversus abdominis; which were better in the respiratory group than in the control group and core group (P < 0.001), and were better in the core group than in the control group (P < 0.001). ConclusionCore training combined with respiratory training can more effectively reduce pain and improve dysfunction by enhancing the strength and control of the core muscles， thus improving the quality of life of patients with lumbar disc herniation.

ObjectiveThe central symptom of Parkinson’s disease (PD) is impaired motor function. Beta-band electrical activity in the motor network of the basal ganglia is closely related to motor function. In this study, we combined scalp electroencephalography (EEG), brain functional network, and clinical scales to investigate the effects of beta burst-period neural electrical activity on brain functional network characteristics, which may serve as a reference for clinical diagnosis and treatment. MethodsThirteen PD patients were included in the PD group, and 13 healthy subjects were included in the healthy control group. Resting-state EEG data were collected from both groups, and beta burst and non-burst periods were extracted. A phase synchronization network was constructed using weighted phase lag indices, and the topological feature parameters of phase synchronization network were compared between the two groups across different periods and four frequency bands. Additionally, the correlation between changes in network characteristics and clinical symptoms was analyzed. ResultsDuring the beta burst period, the topological characteristic parameters of phase synchronization network in all four frequency bands were significantly higher in PD patients compared to healthy controls. The average clustering coefficient of the phase synchronization network in the beta band during the beta burst period was negatively correlated with UPDRS-III scores. In the low gamma band during the non-burst period, the average clustering coefficient of phase synchronization network was positively correlated with UPDRS and UPDRS-III scores, while UPDRS-III scores were positively correlated with global efficiency and average degree. ConclusionThe brain functional network features of PD patients were significantly enhanced during the beta burst period. Moreover, the beta-band brain functional network characteristics during the beta burst period were negatively correlated with clinical scale scores, whereas low gamma-band functional network features during the non-burst period were positively correlated with clinical scale scores. These findings indicate that motor function impairment in PD patients is associated with the beta burst period. This study provides valuable insights for the diagnosis of PD.

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.

Objective:To prepare the anti-programmed death-ligand 1 (PD-L1) nanoantibody P3C8-C3Fab by ligating with C3Fab and to investigate its role in plasma half-life.Methods:The C3Fab peptide derived from protein G was molecularly fused with the nanobody P3C8 by DNA recombination technology. The nanoantibody P3C8-C3Fab was inducibly expressed and purified in the E. coli BL21 strain, and the binding of it to PD-L1 protein, mouse IgG, and PD-L1-expressing tumor cells was detected by enzyme-linked immunosorbent assay (ELISA). The residual P3C8-C3Fab was detected in mouse serum at different times using double-antibody sandwich ELISA to assess the prolongation of the plasma half-life of PD-L1 nanobodies by C3Fab. Results:The nanoantibody P3C8-C3Fab was successfully constructed, and it could efficiently express itself in soluble form in BL21. The purified NbP3C8-C3Fab protein was obtained with a mass fraction of about 90% at a yield of 7.18 mg/L. The affinity of P3C8-C3Fab for PD-L1 protein and mouse IgG gradually increased with increasing mass concentration and showed a concentration correlation. The binding of P3C8-C3Fab to lung cancer A549 cells showed a concentration correlation. The concentration standard curve of P3C8-C3Fab in mouse serum showed a typical S-shape with a concentration correlation. The plasma half-life of P3C8 was only 0.44 h, while the plasma half-life of P3C8-C3Fab was 21.27-fold higher, up to 9.36 h.Conclusions:The linkage of C3Fab to the nanobodies of P3C8 can significantly prolong the plasma half-life of P3C8, which is valuable for the improvement of in vivo nanobody effects.

Objective To explore the effect of robotic training under position limitation on upper limb of patients with shoulder sub-luxation after stroke. Methods From March to December,2023,sixty patients with shoulder subluxation after stroke in the First Affiliated Hospital of Bengbu Medical University were randomized into conventional group(n = 20),robot group(n = 20)and position-limited group(n = 20).All the groups accepted conventional rehabilitation,while the robot group underwent robot training and the position-limited group underwent robot training under position limitation,for four weeks.Before and after treatment,Fugl-Meyer Assessment-Upper Extremities(FMA-UE)and modified Bar-thel Index(MBI)were used for evaluation,surface electromyography(sEMG)was used to measure the average electromyography(AEMG)and root mean square(RMS)of posterior deltoid fasciculus and supraspinatus during contraction,and musculoskeletal ultrasonography was used to determine the distance of acromion-greater tuber-osity(AGT)of the affected side. Results After treatment,FMA-UE score,MBI score,AEMG,RMS and the distance of AGT improved in all the groups(|t|>10.850,P<0.001),and all the indexes were optimal in the position-limited group(F>42.031,P<0.001). Conclusion Robotic training for upper limb under postural limitation can further improve upper limb function and sub-luxation status,activities of daily living,and peri-shoulder muscle in patients with shoulder subluxation after stroke.

Objective:To explore the value of whole exome sequencing for the inferential analysis of recessive genetic disease carrier status for couples with a child died of Primary immunodeficiency (PID).Methods:Clinical data was collected from four couples with a childbearing history of PID who had sought genetic counseling and undergone genetic testing at Henan Provincial People′s Hospital from February 2017 to December 2021. Whole exome sequencing (WES) was performed on both partners of each couple, and candidate variants were validated by Sanger sequencing and fluorescent quantitative PCR. Prenatal diagnosis was conducted on fetuses of these couples after confirming the variants.Results:A total of six variants were detected in four genes including IL2RG, BTK, CYBB, and DUOX2. Among these, the c.1265G>A and c.3329G>A variants of the DUOX2 gene and the c. 676C>T variant of the IL2RG gene were previously known as pathogenic variants. On the other hand, the Exon5_8del variant of the IL2RG gene, the c. 184_185delAC variant of the BTK gene, and the c. 472A>T variant of the CYBB gene were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics, the IL2RG: Exon5_8del, BTK: c. 184_185delAC and CYBB: c. 472A>T variants were classified as likely pathogenic (PVS1+ PM2_Supporting+ PP4).Prenatal diagnosis was conducted for three couples during their subsequent pregnancies, and the results revealed that the fetuses had the wild-type genotypes at the c. 184_185 position of the BTK gene, the c. 472 position of the CYBB gene, and the c. 676 position of the IL2RG gene. Follow-up examinations one year after birth has found no abnormality in the infants. Conclusion:WES is an important tool to infer and analyze the carryier status for couples who had given births to children died of PID and improve the positive detection rate.

Objective:To discuss the effect of Fuzheng Ruanjian Anticancer Formula on the malignant biological behaviors of the hepatocellular carcinoma HepG2 cells by requlating protein kinase B(Akt)/murine double minute 2(MDM2)/P53 signaling pathway.Methods:The HepG2 cells were treated with 0,0.05,0.10,0.20,0.40,0.80,1.60,3.20,and 6.40 g·mL-1 Fuzheng Ruanjian Anticancer Formula for 48 h.CCK-8 method was used to detect the survival rates of the HepG2 cells in various groups,and the concentrations of Fuzheng Ruanjian Anticancer Formula for the subsequent experiments were screened.The HepG2 cells were divided into control group,low dose of Fuzheng Ruanjian Anticancer Formula group(0.2 g·mL-1),medium dose of Fuzheng Ruanjian Anticancer Formula group(0.4 g·mL-1),high dose of Fuzheng Ruanjian Anticancer Formula group(0.8 g·mL-1),SC79 group(8 mg·L-1 SC79),and high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group(0.8 g·mL-1 Fuzheng Ruijian Anticancer Formula+8 mg·L-1 SC79).CCK-8 method was used to detect the proliferation activities of the HepG2 cells in various groups;clone formation assay was used to detect the clone formation rates of the HepG2 cells in various groups;flow cytometry was used to detect the apoptotic rates of the HepG2 cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion HepG2 cells in various groups;Western blotting method was used to detect the expression levels of proliferating cell nuclear antigen(PCNA),cysteine aspartate specific proteinase(Caspase-3),matrix metalloproteinase(MMP)-2,MMP-9,phosphorylated Akt(p-Akt),phosphorylated MDM2(p-MDM2),and P53 proteins in the HepG2 cells in various groups.Results:As the increasing of concentrations of Fuzheng Ruanjian Anticancer Formula(0,0.05,0.10,0.20,0.40,0.80,1.60,3.20,and 6.40 g·mL-1),the surival rates of the HepG2 cells were gradually decreased(P<0.05),and 0.2,0.4,and 0.8 g·mL-1 Fuzheng Ruanjian Anticancer Formula were selected for the subsequent experiments.The CCK-8 assay results showed that compared with control group,the proliferation activities of the HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05),in a dose-dependent manner,while the proliferation activity of the cells in SC79 group was significantly increased(P<0.05).Compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the proliferation activity of the HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased(P<0.05).The clone formation assay results showed that compared with control group,the clone formation rates of the HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the clone formation rate of the cells in SC79 group was significantly increased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the clone formation rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased(P<0.05).The flow cytometry results showed that compared with control group,the apoptotic rates of the HepG2 cells in low,medium,and high doses of Fuzheng Ruijian Anticancer Formula groups were significantly increased(P<0.05)in a dose-dependent manner,while the apoptotic rate of the cells in SC79 group was significantly decreased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the apoptotic rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly decreased(P<0.05).The Transwell chamber assay results showed that compared with control group,the numbers of migration and invasion HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the numbers of migration and invasion cells in SC79 group were significantly increased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the numbers of migration and invasion HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased(P<0.05).The Western blotting results showed that compared with control group,the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the expression levels of Caspase-3 and P53 proteins were significantly increased(P<0.05)in a dose-dependent manner,while the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in SC79 group were significantly increased(P<0.05),and the expression levels of Caspase-3 and P53 proteins were significantly decreased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased(P<0.05),while the expression levels of Caspase-3 and P53 proteins were significantly decreased(P<0.05).Conclusion:Fuzheng Ruanjian Anticancer Formula may inhibit the proliferation,migration,and invasion of the HepG2 cells and promote the apoptosis,and its mechanism may be related to suppressing the Akt/MDM2 signaling pathway and upregulating the P53 proteim expression.

Purpose To investigate the cell components in different tumor lineages of multiple synchronous pituitary neuro-endocrine tumors(PitNETs)/neuroendocrine tumors(MSPs)and to carry out accurate histological typing,which provides an important basis for determining the follow-up plan and adjuvant therapy after surgery.Methods The clinical data of 855 pa-tients with PitNETs were collected and reclassified according to the new WHO standard.The clinicopathological features of 13 patients diagnosed as MSPs were analyzed retrospectively.The immunohistochemical EnVision two-step method was used to de-tect the expression of PIT-1,SF-1,T-PIT,GH,PRL,TSH,LH,FSH,ACTH,etc.,and related literatures were reviewed.Methods A total of 855 cases of pituitary neuroendocrine tumor from the second hospital of the Chinese Hebei Medical U-niversity were collected and reclassified according to the new WHO standard,and review the literature.Results(1)The age of patients were 39-68 years with median age of 55 years.7 cases were female and 6 were male;(2)Imaging findings:There was 1.2～3.8 cm(mean 2.5 cm)in maximum tumor di-ameter.13 patients were large adenomas,neither MSPs nor sin-gle lineage PitNETs could not assessed on imaging;(3)Clinical manifestations:3 cases had hyperprolactinemia which all contai-ning PitNETs components of PRL,one case was immature PIT-1 polyhormone cell tumor,one was dense granular prolactinoma,and one case was eosinophilic stem tumor.One patient had Cushing's disease and contained a Crook cell tumor component;two had elevated ACTH,and one had an adrenocorticotropic ad-enomatous component.In the two patients with no evidence of hormone excess,all contained gonadotropin cell tumor compo-nents;(4)Combination form:11 cases of the combination of the two cell lineages(5 cases of combination of SF-1 lineage and PIT-1 lineage;4 cases of combination of T-PIT lineage and PIT-1 lineage;1 case of null cell tumor and PIT-1 lineage;1 case of plurihormonal PitNETs and SF-1 lineage);Two cases of three cell combinations(null cell tumor,PIT-1 lineage,and T-PIT lineage);Among them,13 cases were PIT-1 lineage tumors,46.2％(6/13)were gonadotropin cell tumor,38.5％(5/13)were prolactinoma;(5)The presence of high-risk lineage tumors in the 10 patient combinations:3 immature PIT-1-lineage tumor,1 Crooke cell PitNETs,1 acidophil stem cells tumor,3 zero-cell PitNETs,and 4 silent-type sparse granular adrenocorti-cotropic hormone PitNETs;Two of them were combinations of two high-risk subtypes.Conclusion MSPs in our center are large adenomas,although their incidence is only 1.5％of Pit-NETs,2/3 cases have high-risk lineage tumor components,and the use of pituitary cell lineage transcription factors and adeno-hypophyseal hormones plays an important role in distinguishing and clarifying the different components of MSPs.