Objective: To investigate the effects of platelet-derived PD-L1 released during storage on T cell proliferation, apoptosis, and cytokine expression. Methods: Apheresis platelets were stored with agitation at 22℃. Platelet supernatant was collected on days 1, 3, 5, and 7 of storage, and then co-cultured with human CD4 and CD8 mixed T cells isolated by magnetic bead sorting. CCK8 assay was used to detect T cell proliferation; flow cytometry was applied to determine T cell apoptosis as well as the expression levels of IFN-γ and GZMB. Additionally, Western blot was employed to examine alterations in downstream signaling molecules of the PD-1 receptor. Results: With the prolongation of platelet storage time, the inhibitory effect of platelet storage medium on T cell proliferation was gradually enhanced, accompanied by suppressed expression of IFN-γ and GZMB, whereas the apoptosis level of T cells increased progressively. Treatment with PD-1 antibody or PD-L1 antibody partially blocked the regulatory effects of platelet storage medium on T cell proliferation, apoptosis, and the expression of IFN-γ and GZMB. Western blot results showed that the phosphorylation levels of p-SHP1 and p-SHP2 were significantly upregulated, whereas p-AKT expression was downregulated in T cells co-cultured with platelet storage medium. Intervention with PD-1 or PD-L1 antibody could partially reverse the above changes in protein expression. Conclusion: During platelet storage, platelet-derived PD-L1 in the storage medium promotes T cell apoptosis and inhibits T cell proliferation as well as the expression of related cytokines in a time-dependent manner. Therefore, platelet-derived PD-L1, as an important immune regulatory mediator, may be involved in transfusion-related immunomodulation (TRIM).

Objective To determine the effect of baicalein on high glucose-induced cardiac fibro-blast pyroptosis based on the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/cysteinyl aspartate-specific proteinase-1(Caspase-1)/gasdermin D(GSDMD)pathway.Methods Rat cardiac fibroblasts were grouped into control,high glucose group,low-,medium-and high-dose baicalein(H-,M-and L-baicalein)groups,and H-baicalein+NLRP3 agonist(BMS-986299)group.Except for the control group,all other groups were cultured in DMEM medium containing 40 mmol/L glucose,then 12.5,25 and 50 μmol/L baicalein was added into the medium correspondingly,and 1 μmol/L BMS-986299 was used to treat the H-baicalein+NLRP3 agonist group.Lactate dehydrogenase(LDH)cytotoxicity assay were employed to detect cell cytotoxicity.qRT-PCR and Western blotting were performed to determine the expression of NLRP3,Caspase-1,and GSDMD at mRNA and protein levels.Results High glucose treatment induced more EdU positive cells,higher pyroptotic rate,stronger cytotoxicity,higher Col-Ⅰ and Col-Ⅲ contents,and enhanced mRNA and protein levels of NLRP3,Caspase-1 and GSDMD in comparison to the control group(P＜0.05).The H-baicalein+NLRP3 agonist group had more EdU positive cells(26.85±2.95 cells vs 15.43±1.82 cells,P＜0.05),higher pyroptotic rate[(33.45±4.02)％vs(17.34±2.15)％,P＜0.05],stronger cytotoxicity[(27.94±2.93)％vs(14.13±1.87)％,P＜0.05],and increased contents of Col-Ⅰ(107.58±13.39 ng/ml vs 58.73±8.36 ng/ml,P＜0.05)and Col-Ⅲ(118.43±13.95 ng/ml vs 68.74±8.57 ng/ml,P＜0.05),and enhanced expression of NLRP3,Caspase-1 and GSDMD at both mRNA and protein levels(P＜0.05)when compared with the H-baicalein group.Conclusion Baicalein inhibits high glucose-induced cardiac fibroblast pyroptosis by suppressing NLRP3/Caspase-1/GSDMD pathway.

AIM To prepare the borneol-menthol eutectic mixture-loaded nanoemulsion gel of tetramethylpyrazine.METHODS The equilibrium solubilities of tetramethylpyrazine in different oil phases,emulsifiers and co-emulsifiers were determined,after which compatibility experiment was performed,and pseudo-ternary phase diagram was drawn.With Km value,oil phase proportion and water phase consumption as influencing factors,particle size,PDI and saturated drug loading as evaluation indices,the formulation was optimized by central composite design-response surface method.The drug-loaded nanoemulsion was dispersed into carbomer 940 gel matrix to prepare nanoemulsion gel,then the physicochemical properties,in vitro drug release and transdermal absorption properties were investigated.RESULTS The optimal formulation was determined to be 3.31∶6.16∶1.56∶88.97 for eutectic mixture-EL-40-1,2-propylene glycol-water ratio,the particle size,PDI and saturated drug loading were 37.85 nm,23.04 and 5.82 mg/g,respectively.The obtained white,semi-solid nanoemulsion gel demonstrated the average pH value and viscosity of 6.68±0.07 and(289.69±1.06)mPa·s,respectively,whose in vitro drug release accorded with Higuchi equation,and the accumulative permeability per unit area was(2 048.23±55.6)μg/cm2 within 24 h,which were 3.72 and 1.21 times higher than those of hydrogel and aqueous solution,respectively.CONCLUSION The borneol-menthol eutectic mixture-loaded nanoemulsion gel of tetramethylpyrazine meets preparation requirements,thus can achieve the effective transdermal delivery of raw medicine.

AIM To investigate the effect of Fuzheng Hefu Zhiyang Formula(FZHFZY)on psoriasis-like skin lesions and immune regulation in mice.METHODS In the in vivo experiment,30 BALB/c mice were randomly divided into the blank group,the model group,the dexamethasone group(1.5 g/kg of compound dexamethasone acetate cream),and the low-dose(2.5 g/kg)and high-dose(5 g/kg)FZHFZY groups,with six mice in each group.The experiment groups were treated with respective FZHFZY and dexamethasone,and the other groups were given normal saline for 10 consecutive days,during which psoriatic skin lesions were induced with imiquimod cream for 7 consecutive days.The mice had their area and severity of psoriasis assessed by PASI score;their histological changes of skin lesions.observed with Hematoxylin-eosin(HE)staining;their F4/80 ratio of skin lesions observed with immunohistochemical(IHC)staining;their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected by Western blot;and their mRNA expressions of tumor necrosis factor-α(TNF-α),IL-17,IL-23 and IL-1β detected by RT-qPCR.In the in vitro research,the cultured RAW264.7 cells were divided into the blank group,the LPS group,and the FZHFZY groups(1 200,600,300,150 μg/mL).The cells had their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected with Western blot;and their mRNA expressions of IL-6,TNF-α,IL-23 and IL-8 detected by RT-qPCR.RESULTS The in vivo experiment showed that compared to the model group,the FZHFZY groups demonstrated decreased PASI score(P＜0.01);improved epidermal thickening and parakeratosis of skin lesions as revealed by HE staining result and increased expression of F4/80 in IHC staining sections;decreased protein expression ratios of p-P38/P38,p-ERK/Erk and p-P65/P65 in skin(P＜0.05,P＜0.01);and reduced mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β in the skin(P＜0.01).FZHFZY(0～2 400 μg/mL)showed no significant cytotoxicity towards RAW264.7 cells in vitro(P＞0.05).Compared to those of the LPS group,the cells exposed to FZHFZ at concentrations of 1 200 and 600 μg/mL demonstrated decreased protein expression ratios of p-P38/P38,p-ERK/Erk,and p-P65/P65(P＜0.05,P＜0.01);and significantly decreased mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β(P＜0.01).CONCLUSION FZHFZY alleviates imiquimod-induced psoriatic lesions in mice and suppresses inflammatory response in LPS-stimulated RAW264.7 cells by inhibiting P38/Erk/NF-κB signaling pathway.

AIM To establish a GC-MS/MS method for the analysis of volatile components in Yinhu Ganmao Powder,and to determine the contents of α-pinene,camphene,sabinene,β-pinene,α-terpinene,(+)-limonene,p-cymene,1,8-cineole,linalool,L-menthol,terpinen-4-ol,DL-menthol,α-terpineol,tridecane,pulegone,caryophyllene,humulene,n-hexadecane and patchouli alcohol.METHODS The analysis was performed on a DB-624 UI capillary column(30 m×0.25 mm×1.40 μm ),and electron ionization source was adopted with multiple reaction monitoring mode.RESULTS Fifty volatile components and twenty-five liposoluble components were identified in volatile oils and medicinal material powder,respectively.Nineteen constituents showed good linear relationships within their own ranges(r ≥ 0.999 0),whose average recoveries were 84.43％-113.31％with the RSDs of less than 9.15％.CONCLUSION This stable,accurate and reproducible method can provide a reference for the quality evaluation of Yinhu Ganmao Powder.

AIM To explore the relieving effect of Er Miao Wan on atopic dermatitis in mice.METHODS In vivo experiment:BALB/c mice were randomly divided into normal group,model group,dexamethasone group(2 mg/kg)and high,medium and low dose groups of Er Miao Wan(4.68,2.34 and 1.17 g/kg).The mouse model of atopic dermatitis was established by repeatedly smearing DNCB solution,and the model was given orally for 21 days.The skin lesion condition on the back of mice,ear swelling degree,and the weight difference between ear lobes were observed and recorded.HE staining was used to observe the histopathological changes in the skin lesion tissues of mice.Toluidine blue(TB)staining was used to observe the infiltration of mast cells in skin lesions.The expression of macrophage marker F4/80 in skin lesions was detected by IHC.The serum levels of TSLP,IL-4,IL-5 and total IgE were detected by ELISA.In vitro experiment:RAW264.7 cells in logarithmic growth period were given 400,200 and 100 μg/mL Er Miao Wan for intervention.Cell proliferation was detected by CCK-8 method.NO level in cell supernatant was detected by Griess method.TNF-α,IL-1 β and IL-6 levels in cell supernatant were detected by ELISA method.The expressions of proteins related to the MAPK/NF-κB signaling pathway in cells was detected by Western blot.RESULTS In vivo experiment:Compared with the model group,the scores of back skin lesions,the swelling degree of right ear and the weight difference between left and right ear pieces in the high-dose group of Er Miao Wan decreased(P＜0.05,P＜0.01),the thickness of skin lesions decreased,the infiltration of mast cells and macrophages decreased(P＜0.05,P＜0.01),and the inflammatory factors TSLP,IL-4,IL-5 and total IgE levels in serum decreased(P＜0.05,P＜0.01),and the expression of F4/80 in the skin lesions decreased(P＜0.01).In vitro experiment:Compared with the model group,the levels of NO,TNF-α,IL-1 β and IL-6 in Er Miao Wan 400 and 200 μg/mL groups decreased(P＜0.05,P＜0.01),and the phosphorylation levels of P38,JNK and P65 proteins decreased(P＜0.05,P＜0.01).CONCLUSION Er Miao Wan can alleviate skin lesion inflammation in DNCB-induced atopic dermatitis mice,and its mechanism may be related to inhibiting the activation of MAPK/NF-κB signaling pathway of macrophage,reducing macrophage infiltration and reducing Th2 cytokines.

The telomere structure in the cell nucleus is crucial for maintaining the stability and functions of chromosomes.Telomerase is a ribonucleoprotein reverse transcriptase,which catalyzes the elongation of telomeres using its own RNA as a template,thereby counteracting the shortening of telomeres caused by chromosome replication and cell division.Due to its overexpression in over 85％of malignant tumor cells,telomerase has emerged as a highly promising biomarker and a novel target for cancer therapy.In recent years,given the importance of precise quantification of telomerase activity in guiding medical diagnosis and treatment strategies,researchers have developed various high-performance telomerase detection techniques.Among these,electrochemical biosensing technique has cause much attention due to its high sensitivity,operational convenience,rapid response,and ease of miniaturization.This paper focused on the latest advances in electrochemical sensing technique for detection of telomerase activity,aiming to provide inspiration for designing novel telomerase activity detection strategies by elucidating three unique properties of telomerase primer extension products.

Haloacetic acids(HAAs),as a class of disinfection byproducts in drinking water,pose potential threats to human health,so the rapid,accurate and simultaneous detection of HAAs is of great significance for ensuring drinking water safety.Aiming at the challenges in HAAs detection and risk analysis,a novel method for synchronous rapid detection of seven kinds of HAAs in drinking water based on in situ derivatization technology and headspace gas chromatography was developed in this study.Through single-factor optimization experiments,the optimal reaction parameters for in situ derivatization were determined,including the type and dosage of salting-out agent,the acidity of reaction system,the amount of phase transfer catalyst,the dosage of derivatization agent,and the extraction solvent volume.Methodologic validation showed that the seven kinds of HAAs exhibited excellent linear relationships within their respective detection concentration ranges(R2>0.998).The method detection limits(MDLs)ranged from 0.04 to 0.33 μg/L,and the limits of quantification(LOQs)were between 0.14 and 1.34 μg/L.For real water samples,the average spiked recoveries of the seven HAAs ranged from 90.9%to 107.7%,with relative standard deviation(RSDs)between 1.55%and 6.49%,and the HAAs contents in all tested samples were below the limits specified in the Standards for Drinking Water Quality(GB 5749-2022)of China.This method was featured with simple operation,fast analysis speed,high sensitivity,and good accuracy,providing an efficient and reliable technical support for routine monitoring of HAAs contaminants in drinking water and showing promising application value for widespread promotion.

Objective To investigate the clinical effect of percutaneous injection of calcitonin and methylprednone in the treatment of aneurysmal bone cyst in children.Methods From June 2020 to March 2023,19 patients with aneurysmal bone cysts were admitted and all were treated with percutaneous puncture injection of calcitonin and methylprednisolone.After percutaneous puncture biopsy,200 iu of calcitonin and 120 mg of methylprednone were injected into the cyst of aneurysmal bone cyst.Compare the volume of the cyst cavity and the thinnest cortical bone thickness around the cyst cavity before and after the operation.All patients were followed up,while preoperative and postoperative tumor cavity and the thinnest peritumor cortical thickness were compared.Results 19 patients were followed up for an average of 16 months(12-52 months).Of the 19 patients,17 were effective,the effective rate was 89.5％,and no serious complications occurred in all cases.At the last follow-up,the volume of the tumor cavity was significantly lower than that before surgery[(37.05±21.17)cm3,(110.95±45.67)cm3],and the difference was statistically significant(P＜0.05).The average thinnest cortical bone thickness around the tumor cavity was(0.71±0.37)mm before surgery and(1.87±0.60)mm after surgery.The difference was statistically significant(P＜0.05).Conclusion Percutaneous injection of calcitonin and methylprednone in the treatment of aneurysmal bone cysts in children is minimally invasive,safe and effective.It can be used as the first-line treatment for aneurysmal bone cyst in children.

Objective To evaluate the predictive value of dual-modality ultrasound,incorporating conventional ultrasound and ultrasound elastography,in combination with S-Detect for cervical lymph node metastasis(CLNM)in patients with papillary thyroid carcinoma(PTC).Methods A retrospective analysis was conducted on the clinical data of 135 patients diagnosed with PTC who received treatment at the First Affiliated Hospital of Shihezi University between November 2023 and August 2024.For all patients,clinical baseline characteristics,conventional ultrasound findings,ultrasound elastography results,and S-Detect analysis data were collected.Independent predictors of CLNM in PTC were identified,and predictive models were developed.Receiver operating characteristic(ROC)curves were generated to compare the area under the curve(AUC)of the models.The most effective predictive model was selected to construct a risk probability nomogram,and the predictive performance and clinical applicability of this nomogram were subsequently evaluated.Results Age,maximum nodule diameter,boundary characteristics,capsular invasion,transverse-sectional morphological findings assessed by S-Detect,and ECI-based elasticity grading were identified as independent predictors of CLNM in PTC(all P<0.05).The AUC of the predictive model constructed using these six variables was 0.890(95%CI:0.835～0.945).The calibration curve demonstrated strong agreement between predicted and observed outcomes,and decision curve analysis indicated that the nomogram provided a favorable net clinical benefit within a threshold probability range of 2%to 91.5%.Conclusions Age,maximum nodule diameter,boundary characteristics,capsular invasion,sonographic features assessed by S-Detect in the transverse plane,and ECI-based elasticity grading are independent predictors of CLNM in PTC.A nomogram model incorporating these parameters demonstrates effective performance in predicting the likelihood of CLNM.