Objective: To investigate the effects of clinical routine X-ray irradiation dose (average irradiation dose: 29.7±0.54 Gy) on the function, apoptosis, activation state and mitochondrial function of platelets during in vitro storage, so as to provide experimental evidence for optimizing platelet irradiation strategies. Methods: A paired experimental design was adopted. Platelets were collected from 12 healthy donors, and each sample was equally divided into the irradiated group and the control group (non-irradiated). All samples were stored for 5 days under standard platelet preservation conditions (22±2℃, continuous oscillation). Flow cytometry was used to detect platelet count, apoptosis rate (Annexin V+ positive rate), activation markers (CD62P, PAC-1, CD42b) and reactive oxygen species (ROS) level. Meanwhile, mitochondrial-specific probes were used to evaluate changes in mitochondrial count, membrane potential and adenosine triphosphate (ATP) content. Additionally, transmission electron microscopy (TEM) was employed to observe the ultrastructure of platelets, with a focus on mitochondrial morphology, platelet membrane integrity and granule distribution. Results: Within 5 days of storage, the platelet count was (841±89.16)×10 /L in the irradiated group and (824.5±92.88)×10 /L in the control group, with no statistically significant difference between the two groups (P=0.54). The apoptosis rate was (4.94±1.39) % in the irradiated group and (5.50±0.83) % in the control group, showing no significant difference (P=0.31). For activation indicators, the CD62P expression rate was (24.32±7.57) % in the irradiated group versus (25.21±8.13) % in the control group (P=0.43). The PAC-1 positive rates were (12.15±4.43) % and (11.75±3.40) % in the irradiated group and control group, respectively (P=0.44). The CD42b expression rates were (12.14±4.43) % and (11.75±3.4) % in the two groups, respectively (P=0.47). The ROS levels were (31.98±8.1) % and (30.64±5.89) % in the two groups, respectively (P=0.45). No significant differences were found in the above indicators. For mitochondrial function indicators, the mitochondrial count was (55.88±11.49) % in the irradiated group and (53.5±7.24) % in the control group (P=0.57). The ATP contents were (42.45±5.29) % and (41.58±9.50) % in the irradiated group and control group, respectively (P=0.77). The relative membrane potential values were (59.53±10.89) % and (57.49±6.54) % in the two groups, respectively (P=0.47). No significant difference were observed on the mitochondrial function-related indicators. TEM further confirmed that the ultrastructure of platelets in the irradiation group was intact, the mitochondrial morphology was normal, and no pathological changes such as swelling or vacuolization were observed. Conclusion: This study evaluated the impact of conventional-dose X-ray irradiation on platelet storage quality, confirming that this dose does not significant impair platelet count, apoptosis rate, activation status, or mitochondrial function. This finding provides important experimental evidence for the clinical promotion of X-ray irradiation technology and suggests its potential as a safe alternative to γ irradiation. Future studies could further expand the sample size and extend the observation period to verify the effects of X-ray irradiation on long-term platelet storage and post-transfusion in vivo survival rate.

Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.

Objective:To investigate the effects of Yisui Shengxue Decoction on the regulation of Notch signaling pathway on bone marrow hematopoiesis in mice with aplastic anemia(AA).Methods:A total of 60 CByB6F1 mice were randomly divided into blank group,model group,positive control cyclosporine(CsA)group,low-dose(YSSX-L)group and high-dose(YSSX-H)group of Chinese herbal compound Yisui Shengxue Decoction,with 12 mice in each group.Except for the blank group,the mice in each group were given 5 Gy X-ray irradiation combined with tail vein infusion of lymphocytes to establish the immune-mediated AA mouse model.The low and high dose groups were given 8.6 and 17.2 g/(kg?d)of Yisui Shengxue Decoction by gavage,the model group and the blank group were given equal volume of saline by gavage,and the positive control group was given 25 mg/(kg?d)of cyclosporine by ga-vage for 14 days.The mice were tested for white blood cell count(WBC),red blood cell count(RBC),hemoglobin(Hb),platelet count(PLT)and bone marrow nucleated cell count in peripheral blood,bone marrow pathological changes and the levels of IFN-γ,IL-4 and IL-5 in peripheral serum,and T-lymphocyte transcription factors T-bet,GATA3,RORγt,FOXP3 mRNA and protein in spleen tissues,as well as the expression levels of Notch signaling pathway-related genes and protein in spleen tissues.Results:Compared with the blank group,the peripheral blood WBC,RBC,Hb,PLT,bone marrow nucleated cell count,expressions of serum IL-4,IL-5 and spleen tissue GATA3,FOXP3 mRNA and protein were significantly lower in the model group(P<0.01),and the expressions of serum IFN-γ,spleen tissue T-bet,RORγt mRNA and protein,and Notch1,Jagged1,DLL4 mRNA and protein were significantly in-creased(P<0.05).Compared with the model group,peripheral blood WBC,RBC,Hb,PLT,bone marrow nucleated cell count,ex-pressions of serum IL-4,IL-5 and spleen tissue GATA3,FOXP3 mRNA and protein were significantly higher in each intervention group(P<0.05),and the expression of serum IFN-γ and spleen tissue T-bet,RORγt mRNA and protein as well as Notch1,Jagged1,DLL4 mRNA and protein were significantly reduced(P<0.05).Conclusion:Yisui Shengxue Decoction can improve bone marrow he-matopoietic function and regulate immune disorders in AA mice,and its mechanism of action may be related to the regulation of Notch signaling pathway.

Objective:To establish ICP-MS method for determining the Class 1 and Class 2A elemental impurities in pharmaceutical excipients polysorbates according to ICH Q3D(R2)Guidelines.Class 1 elemental impurities include As,Cd,Hg and Pb;Class 2A elemental impurities include Co,Ni and V.Methods:The analysis was performed using ICP-MS with the following parameters.Plasma mode:HMI-8,carrier gas:He,sampling depth:10.0 mm.RF power:1 600 W,the carrier gas flow,the nebulizer gas flow and the auxiliary gas flow were all 0.8 L·min-1.The plasma gas flow was 15.0 L·min-1.The speed of nebulizer pump was0.10 r·min-1.The temperature of atomizer chamber was 2.0℃.Analytical method:internal standard method.Results:The correla-tion coefficient for all elemental impurities was not less than 0.999.Spiked recoveries ranged between 85%-115.0%,with RSD is not more than 5.0%.The RSD of precision rate was not more than 10.0%.The contents of 7 elemental impurities in 15 batches of the samples were all less than PDE for injection set by ICH Q3D(R2)guidelines.Conclusion:The developed ICP-MS method demonstrates high sensitivity,good precision,and accuracy,making it suitable for controlling the Class 1 and Class 2A elemental impurities in polysorbate pharma-ceutical excipients.The results shows low risks of elemental impurities in commercial pharmaceutical excipients polysorbates.

Objective:To establish ICP-MS method for determining the Class 1 and Class 2A elemental impurities in pharmaceutical excipients polysorbates according to ICH Q3D(R2)Guidelines.Class 1 elemental impurities include As,Cd,Hg and Pb;Class 2A elemental impurities include Co,Ni and V.Methods:The analysis was performed using ICP-MS with the following parameters.Plasma mode:HMI-8,carrier gas:He,sampling depth:10.0 mm.RF power:1 600 W,the carrier gas flow,the nebulizer gas flow and the auxiliary gas flow were all 0.8 L·min-1.The plasma gas flow was 15.0 L·min-1.The speed of nebulizer pump was0.10 r·min-1.The temperature of atomizer chamber was 2.0℃.Analytical method:internal standard method.Results:The correla-tion coefficient for all elemental impurities was not less than 0.999.Spiked recoveries ranged between 85%-115.0%,with RSD is not more than 5.0%.The RSD of precision rate was not more than 10.0%.The contents of 7 elemental impurities in 15 batches of the samples were all less than PDE for injection set by ICH Q3D(R2)guidelines.Conclusion:The developed ICP-MS method demonstrates high sensitivity,good precision,and accuracy,making it suitable for controlling the Class 1 and Class 2A elemental impurities in polysorbate pharma-ceutical excipients.The results shows low risks of elemental impurities in commercial pharmaceutical excipients polysorbates.

Objective:To investigate the effects of Yisui Shengxue Decoction on the regulation of Notch signaling pathway on bone marrow hematopoiesis in mice with aplastic anemia(AA).Methods:A total of 60 CByB6F1 mice were randomly divided into blank group,model group,positive control cyclosporine(CsA)group,low-dose(YSSX-L)group and high-dose(YSSX-H)group of Chinese herbal compound Yisui Shengxue Decoction,with 12 mice in each group.Except for the blank group,the mice in each group were given 5 Gy X-ray irradiation combined with tail vein infusion of lymphocytes to establish the immune-mediated AA mouse model.The low and high dose groups were given 8.6 and 17.2 g/(kg?d)of Yisui Shengxue Decoction by gavage,the model group and the blank group were given equal volume of saline by gavage,and the positive control group was given 25 mg/(kg?d)of cyclosporine by ga-vage for 14 days.The mice were tested for white blood cell count(WBC),red blood cell count(RBC),hemoglobin(Hb),platelet count(PLT)and bone marrow nucleated cell count in peripheral blood,bone marrow pathological changes and the levels of IFN-γ,IL-4 and IL-5 in peripheral serum,and T-lymphocyte transcription factors T-bet,GATA3,RORγt,FOXP3 mRNA and protein in spleen tissues,as well as the expression levels of Notch signaling pathway-related genes and protein in spleen tissues.Results:Compared with the blank group,the peripheral blood WBC,RBC,Hb,PLT,bone marrow nucleated cell count,expressions of serum IL-4,IL-5 and spleen tissue GATA3,FOXP3 mRNA and protein were significantly lower in the model group(P<0.01),and the expressions of serum IFN-γ,spleen tissue T-bet,RORγt mRNA and protein,and Notch1,Jagged1,DLL4 mRNA and protein were significantly in-creased(P<0.05).Compared with the model group,peripheral blood WBC,RBC,Hb,PLT,bone marrow nucleated cell count,ex-pressions of serum IL-4,IL-5 and spleen tissue GATA3,FOXP3 mRNA and protein were significantly higher in each intervention group(P<0.05),and the expression of serum IFN-γ and spleen tissue T-bet,RORγt mRNA and protein as well as Notch1,Jagged1,DLL4 mRNA and protein were significantly reduced(P<0.05).Conclusion:Yisui Shengxue Decoction can improve bone marrow he-matopoietic function and regulate immune disorders in AA mice,and its mechanism of action may be related to the regulation of Notch signaling pathway.

Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.

ObjectiveTo investigate the therapeutic effect of Baihe Wuyaotang （BWT） on non-alcoholic fatty liver disease （NAFLD） and elucidate its underlying mechanism. MethodC57BL/6J mice were randomly assigned to six groups： normal control， model， positive drug （pioglitazone hydrochloride 1.95×10^-3 g·kg^-1）， and low-， medium-， and high-dose BWT （1.3，2.5 and 5.1 g·kg^-1）. Following a 12-week high-fat diet （HFD） inducement， the mice underwent six weeks of therapeutic intervention with twice-daily drug administration. Body weight was monitored weekly throughout the treatment period. At the fifth week， glucose tolerance （GTT） and insulin tolerance （ITT） tests were conducted. Subsequently， the mice were euthanized for the collection of liver tissue and serum， and the subcutaneous adipose tissue （iWAT） and epididymal adipose tissue （eWAT） were weighed. Serum levels of total triglycerides （TG） and liver function indicators，such as alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， were determined. Histological examinations， including oil red O staining， hematoxylin-eosin （HE） staining， Masson staining， and transmission electron microscopy， were performed to evaluate hepatic lipid deposition， pathological morphology， and ultrastructural changes， respectively. Meanwhile， Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were employed to analyze alterations， at both gene and protein levels， the insulin signaling pathway molecules， including insulin receptor substrate 1/2/protein kinase B/forkhead box gene O1 （IRS1/2/Akt/FoxO1）， glycogen synthesis enzymes phosphoenolpyruvate carboxy kinase （Pepck） and glucose-6-phosphatase （G6Pase）， lipid metabolism-related genes stearoyl-coA desaturase-1 （SCD-1） and carnitine palmitoyltransferase-1 （CPT-1）， fibrosis-associated molecules α-smooth muscle actin （α-SMA）， type Ⅰ collagen （CollagenⅠ）， and the fibrosis canonical signaling pathway transforming growth factor-β₁/drosophila mothers against decapentaplegic protein2/3（TGF-β₁/p-Smad/Smad2/3）， inflammatory factors such as interleukin（IL）-6， IL-8， IL-11， and IL-1β， autophagy markers LC3B Ⅱ/Ⅰ and p62/SQSTM1， and the expression of mammalian target of rapamycin （mTOR）. ResultCompared with the model group， BWT reduced the body weight and liver weight of NAFLD mice（P<0.05， P<0.01）， inhibited liver lipid accumulation， and reduced the weight of white fat： it reduced the weight of eWAT and iWAT（P<0.05， P<0.01） as well as the serum TG content（P<0.05， P<0.01）. BWT improved the liver function as reflected by the reduced ALT and AST content（P<0.05， P<0.01）. It improved liver insulin resistance by upregulating IRS2， p-Akt/Akt， p-FoxO1/FoxO1 expressions（P<0.05）. Besides， it improved glucose and lipid metabolism disorders： it reduced fasting blood glucose and postprandial blood glucose（P<0.05， P<0.01）， improved GTT and ITT（P<0.05， P<0.01）， reduced the expression of Pepck， G6Pase， and SCD-1（P<0.01）， and increased the expression of CPT-1（P<0.01）. The expressions of α-SMA， Collagen1， and TGF-β₁ proteins were down-regulated（P<0.05， P<0.01）， while the expression of p-Smad/Smad2/3 was downregulated（P<0.05）， suggesting BWT reduced liver fibrosis. BWT inhibited inflammation-related factors as it reduced the gene expression of IL-6， IL-8， IL-11 and IL-1β（P<0.01） and it enhanced autophagy by upregulating LC3B Ⅱ/Ⅰ expression（P<0.05）while downregulating the expression of p62/SQSTM1 and mTOR（P<0.05）. ConclusionBWT ameliorates NAFLD by multifaceted improvements， including improving IR and glucose and lipid metabolism， anti-inflammation， anti-fibrosis， and enhancing autophagy. In particular， BWT may enhance liver autophagy by inhibiting the mTOR-mediated signaling pathway.

Objective To explore the relationship between serum microRNA-124(miR-124),CD146,angio-poietin-like protein 2(Angptl2)and the stability of carotid atherosclerosis(CAS)plaque in patients with a-cute cerebral infarction(ACI),and to provide reference for early prevention and treatment of patients with ACI.Methods A total of 191 patients with ACI admitted in Handan Central Hospital from January 2020 to February 2023 were selected as ACI group,and another 61 healthy volunteers who were underwent physical examination during the same period were selected as control group.The patients with ACI were divided into unstable plaque group(56 cases),stable plaque group(71 cases),and non plaque group(64 cases)based on carotid color doppler ultrasound results.The serum miR-124 expression levels of all subjects were detected by real time fluorescence quantitative polymerase chain reaction(RT-qPCR),and the serum CD146 and Angptl2 levels were detected by enzyme-linked immunosorbent assay(ELISA).The influencing factors of the instabili-ty of CAS plaque in patients with ACI was analyzed by univariate and multivariate Logistic regression.The predictive value of serum miR-124,CD146 combined with Angptl2 for the instability of CAS plaque in patients with ACI was analyzed by the receiver operating characteristic(ROC)curve.Results The serum CD146 and Angptl2 levels in ACI group were higher than those in control group(P＜0.05),and the miR-124 expression level was lower than that in control group(P＜0.05).Univariate analysis showed that the stability of CAS plaques in ACI patients was correlated with age,hypertension,hyperlipidemia,fibrinogen(FIB),serum C-reac-tive protein(CRP),serum cystatin C(CyC),CD146,Angptl2 and miR-124(P＜0.05).Multivariate Logistic regression analysis showed that the decrease of serum miR-124,the increase of CD146,the increase of Angptl2 and the combination of hyperlipidemia were risk factors for CAS plaque stability in ACI patients(P＜0.05).The area under ROC curve(AUC)of serum miR-124,CD146,Angptl2 and the combination of the three indi-cators to predict CAS plaque instability in ACI patients were 0.741,0.719,0.781 and 0.834,respectively.Conclusion The serum miR-124 expression level,CD146 and Angptl2 levels are the influencing factors of CAS plaque instability in ACI patients,which may be involved in the formation and development of CAS plaque in ACI patients.The combined detection of the three factors has a good predictive effect on CAS plaque instability in ACI patients.

Objective:To determine the erythromycin resistance of Bordetella pertussis isolates and their ptxP1 and ptxP3 phenotypic composition and compare clinical manifestations of children with pertussis caused by the two types of strains. Methods:This was a cross-sectional study, the pertussis cases diagnosed using bacterial culture from January 2019 to December 2022 in Beijing Children′s Hospital and the First People′s Hospital of Wuhu were collected.Any suspected Bordetella pertussis colonies were identified by the slide agglutination test.The susceptibility of isolates to erythromycin was detected by the E-test and K-B test.The ptxP gene was amplified by polymerase chain reaction and sequenced to determine its genotype. t-test, Mann-Whitney U-test, Chi-square test and Fisher′s exact test were use to statistical analysis. Results:A total of 192 strains of Bordetella pertussis were identified, including 188 (97.9%) erythromycin-resistant strains.Among the 188 strains, 30.3%(57/188) belonged to the ptxP1 genotype and 69.7%(131/188) belonged to the ptxP3 genotype.In children aged below 1 year old, the incidence of paroxysmal cough caused by infection with the ptxP3 strain was higher than that with the ptxP1 strain (57.1% vs.29.4%, P<0.05), and children infected with the ptxP3 strain were more likely to develop apnea or asphyxia (23.8% vs.17.6%), post-tussive vomiting (44.4% vs.32.4%), whooping cough (72.0% vs.50.0%) and pneumonia or bronchitis (85.7% vs.73.5%) compared to those infected with the ptxP1 strain, but the differences were not statistically significant(all P>0.05). In children aged 1 year old and above, the white blood cell count of children infected with the ptxP1 strain was higher than that of infections with the ptxP3 strain [13.5(9.9, 24.5)×10 9/L, 10.3 (7.0, 16.4)×10 9/L, P<0.05], and children infected with the ptxP1 strain were more likely to contract other pathogen infections than those infected with the ptxP3 strain (17.4% vs.4.4%, P>0.05). Conclusions:ptxP3 erythromycin-resistant Bordetella pertussis has become the main pathogen of pertussis.Infants with pertussis caused by the ptxP3 erythromycin-resistant strain show more significant manifestations and a higher possibility of severe symptoms than those infected with the ptxP1 erythromycin-resistant strain.