Aim To establish the pyroptosis model of rat chondrocytes induced by tumor necrosis factor α(TNF-α)in order to study the effect of lysine acetyl-transferase 7(KAT7)on pyroptosis of chondrocytes.Methods Chondrocytes of rat knee joint were isolated by type Ⅱ collagenase digestion,and were identified by toluidine blue staining and Col Ⅱ immunofluorescence.CCK-8 was used to evaluate cell viability.Western blot was used to detect the expression of pyroptosis-related proteins NLRP3,GSDMD,caspase-8 and KAT7 in cells intervened with TNF-α,adenovirus overexpression of KAT7(KAT7-oe)and KAT7 inhibitor WM-3835.The microstructure of the cells was observed by scanning e-lectron microscopy.Pyroptosis was detected by TUNEL staining,and the expression of pyroptosis-related pro-tein and KAT7 was detected by immunofluorescence.Results Compared with the empty virus group,KAT7-oe inhibited cell viability,promoted the expression of pyroptosis-related proteins,and TNF-α enhanced this effect.At the same time,the expression of KAT7 and pyroptosis-related proteins in the TNF-α stimulation group increased,and WM-3835 reduced the related proteins expression.Electron microscopy showed that KAT7-oe caused cell swelling,deformation,membrane perforation and rupture,while WM-3835 could restore cell morphology.TUNEL staining and immunofluores-cence results also confirmed that KAT7-oe induced chondrocyte pyroptosis,and WM-3835 could down-reg-ulate the fluorescence of pyroptosis-related proteins.Conclusions The expression of KAT7 increases in rat chondrocyte pyroptosis model,and the intervention of KAT7 expression affects signal molecules related to py-roptosis pathway,suggesting that KAT7 may be related to chondrocyte pyroptosis.

Aim To establish the pyroptosis model of rat chondrocytes induced by tumor necrosis factor α(TNF-α)in order to study the effect of lysine acetyl-transferase 7(KAT7)on pyroptosis of chondrocytes.Methods Chondrocytes of rat knee joint were isolated by type Ⅱ collagenase digestion,and were identified by toluidine blue staining and Col Ⅱ immunofluorescence.CCK-8 was used to evaluate cell viability.Western blot was used to detect the expression of pyroptosis-related proteins NLRP3,GSDMD,caspase-8 and KAT7 in cells intervened with TNF-α,adenovirus overexpression of KAT7(KAT7-oe)and KAT7 inhibitor WM-3835.The microstructure of the cells was observed by scanning e-lectron microscopy.Pyroptosis was detected by TUNEL staining,and the expression of pyroptosis-related pro-tein and KAT7 was detected by immunofluorescence.Results Compared with the empty virus group,KAT7-oe inhibited cell viability,promoted the expression of pyroptosis-related proteins,and TNF-α enhanced this effect.At the same time,the expression of KAT7 and pyroptosis-related proteins in the TNF-α stimulation group increased,and WM-3835 reduced the related proteins expression.Electron microscopy showed that KAT7-oe caused cell swelling,deformation,membrane perforation and rupture,while WM-3835 could restore cell morphology.TUNEL staining and immunofluores-cence results also confirmed that KAT7-oe induced chondrocyte pyroptosis,and WM-3835 could down-reg-ulate the fluorescence of pyroptosis-related proteins.Conclusions The expression of KAT7 increases in rat chondrocyte pyroptosis model,and the intervention of KAT7 expression affects signal molecules related to py-roptosis pathway,suggesting that KAT7 may be related to chondrocyte pyroptosis.

Objective To obtain the monitoring measurement range of quality indicators of red blood cells,plasma and derivatives and leukocyte-reduced apheresis platelets provided by blood stations in Hebei province,explore the distribution of monitoring values and the change of monitoring level,so as to further strengthen the homogenization construction of quality control laboratories in blood stations in Hebei.Methods In 2023,the sampling data of 12 blood stations in Hebei from 2015 to 2022 were collected,scatter plots were made and the range markers were set,and the"mean±SD"line was taken as the upper limit and lower limit of the measurement range.In 2024,the monitoring values in 2023 were added,and the changes of two measurement ranges were compared to analyze the stability and overall level.Results Comparison of the measurement range from 2015 to 2022 and the measurement range from 2015 to 2023 showed that the standard deviation of the content of deleukocyte suspension of red blood cells-hemoglobin,washed erythrocyte-hemoglobin,washed erythrocyte-su-pernatant protein,cryoprecipitate coagulation factor-FⅧ,fresh frozen plasma-FⅧ,leukocyte-reduced apheresis platelets-leukocyte residue and leukocyte-reduced apheresis platelet-red blood cell concentration decreased from 8.132 to 7.993,6.252 to 6.104,0.273 to 0.267,57.506 to 56.276,0.920 to 0.892,0.653 to 0.644 and 2.653 to 2.603,respectively.The narrowing of the standard deviation range of the above items led to more concentrated monitoring values and reduced disper-sion.Comparison of the measurement range from 2015 to 2022 and the measurement range from 2015 to 2023 showed that the mean value of leukocyte residue of the deleukocyte suspension of red blood cells,hemoglobin content of the wash eryth-rocyte,protein content of supernatant of the wash erythrocyte,hemolysis rate of the wash erythrocyte,FⅧ content of the cryoprecipitate coagulation factor,plasma protein content of the fresh frozen plasma,FⅧ content of the fresh frozen plasma,platelet content of the leukocyte-reduced apheresis platelets changed from 0.362 to 0.476,44.915 to 44.861,0.280 to 0.283,0.137 to 0.142,133.989 to 133.271,60.262 to 60.208,1.301 to 1.277 and 3.036 to 3.033,respectively,and was closer to the national standard line,which reflects an increase in the number of unqualified monitoring values or values close to the national standard line in 2023.The long-term qualified rate of coagulation items was low,and no improvement has been ob-served.The stability of biochemical items has been enhanced but overall deviation has occurred,with the average value close to the national standard line.The possibility of subsequent testing failure has increased.The counting items showed no obvi-ous common characteristics.Conclusion The use of"mean±SD"in the analysis can visually display the distribution of mo-nitoring values of different items in Hebei,forming an indicator measurement range covering the past nine years.It shows the characteristics of each item,and provides reference for subsequent quality control laboratory data analysis of each blood sta-tions to takes active measures to improve the monitoring level.

Hydrogel is a kind of material with high water content,good biocompatibility and extracellular matrix-like property,among which polypyrrole(PPy)conductive hydrogels have both physical characteristics and excellent conductivity of hydrogels themselves.Its conductivity can be used to detect electrical signals generated in biological systems and provide electrical stimulation to regulate the activities and functions of cells and tissues.These characteristics make it widely used in the biomedical field.The recent progress of PPy conductive hydrogels in biomedical field was reviewed in this paper.In terms of classification,according to the cross-linking mechanism of PPy and hydrogel matrix,the non-covalent cross-linked PPy conductive hydrogels and covalent cross-linked PPy conductive hydrogels were divided.The applications of PPy conductive hydrogels in the biomedical field(Skin damage repair,nerve repair,myocardial repair and flexible sensing,etc.)were mainly introduced,and the development trend and challenges of PPy conductive hydrogels in the biomedical field were discussed.

Remitting seronegative symmetrical synovitis with pitting edema (RS3PE) syndrome is a rare seronegative synovial inflammatory disease in which fever is a rare symptom.There are few case reports of RS3PE syndrome with fever as the first clinical manifestation in China.In this paper,we report a case of RS3PE syndrome with fever as the first symptom and diagnosed by systematic fever investigation.
Humans ; Edema/etiology* ; Fever/etiology* ; Syndrome ; Synovitis/drug therapy*

【Objective】 To conduct the laboratory quality assessment between 12 blood stations in Hebei province, analyze the results and explore the accuracy and comparability of testing, so as to improve the level of testing ability and quality management. 【Methods】 With reference to the external quality assessment rules of National Center for Clinical Laboratories and combined with the instructions of quality assessment samples, daily testing process of the laboratories were assessed. The quality indicators include blood cell count (WBC, RBC, Hb, HCT, MCV, MCH, MCHC and PLT), biochemical items (TP) and coagulation parameters (FIB and FⅧ). 【Results】 There are still problems in laboratories in terms of personnel operation, instrument maintenance and the impact of different reagent batches, especially in biochemical items and coagulation parameters. The pass rate of biochemical items was the lowest, only 72.75%, and that of blood cell count was the highest, reaching 98.75%. 【Conclusion】 With the progress of the project, the quality monitoring level of daily blood sampling tests in the quality control laboratory of each blood station has been improved. However, it is still necessary for each laboratory to improve the testing ability and quality management to a higher level in Hebei.

This study aims to explore the effect of butyl alcohol extract of Baitouweng Decoction(BAEB) on vulvovaginal candidiasis(VVC) in mice and to clarify the mechanism from Toll-like receptors(TLRs)/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome. To be specific, female KM mice were randomized into control group(i.g., normal saline), model group, fluco-nazole group(i.g., 20 mg·kg~(-1)), and low-dose, medium-dose, and high-dose BAEB groups(i.g., 20, 40, and 80 mg·kg~(-1), respectively). VVC was induced in mice except the control group. After the modeling, administration began and lasted 7 days. The ge-neral conditions and body weight of mice were recorded every day. On the 1 st, 3 rd, 7 th, and 14 th after vaginal infection by Candida albicans, the fungal load in the vaginal lavage fluid of the mice was measured with the plate method, and the morphology of C. albicans in vaginal lavage fluid was observed based on Gram staining. After the mice were killed, vaginal tissues were subjected to hematoxylin-eosin(HE) staining and periodic acid-Schiff(PAS) staining for vaginal histopathological analysis. The content of cytokines in vaginal lavage fluid, such as interleukin(IL)-1β, IL-18, tumor necrosis factor-α(TNF-α), IL-6, and S100 a8, was determined by enzyme-linked immunosorbent assay(ELISA), and content of reactive oxygen species(ROS) in vaginal tissues by tissue ROS detection kit. The protein expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and nuclear factor-κB(NF-κB) in vaginal tissues was detected by Western blot, and the levels and distribution of NLRP3, Dectin-1, Syk, MyD88, TLR2, and TLR4 in vaginal tissues were determined with the immunohistochemical method. The results show that BAEB can improve the general conditions of VVC mice, reduce the fungal load and C. albicans hyphae in vaginal secretion, decrease ROS content in vaginal tissues and content of cytokines in vaginal lavage fluid, and down-regulate the expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and NF-κB in vaginal tissues. The above results indicate that BAEB exerts therapeutic effect on VVC mice by down-regulating the key proteins in the TLRs/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome.
1-Butanol/therapeutic use* ; Animals ; Candida albicans ; Candidiasis, Vulvovaginal/drug therapy* ; Caspase 1/metabolism* ; Cytokines/metabolism* ; Female ; Humans ; Inflammasomes/metabolism* ; Mice ; Myeloid Differentiation Factor 88/metabolism* ; NF-kappa B/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Plant Extracts/therapeutic use* ; Reactive Oxygen Species/metabolism* ; Signal Transduction ; Toll-Like Receptor 2/metabolism* ; Toll-Like Receptor 4/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.

Objective:To explore the injury effect and its possible mechanism of amiodarone on human umbilical vein endothelial cells (HUVECs).Methods:After 3 generations of cultivation, the HUVECs were seeded in 96-well plates and incubated with amiodarone (0, 10, 20, 30, and 60 μmol/L) for 24 hours. The cell viability was detected using cell counting kit 8 (CCK-8) assay and the relative viability of cells incubated with different concentrations of amiodarone were calculated by taking the cell viability of the 0 μmol/L group as 100%. The concentration of amiodarone at which cell viability was reduced to 70% was selected for subsequent experiments. The effect of amiodarone of this concentration on the activity of HUVECs after action for different time (6, 12, 24, 36, and 48 hours) was detected using the CCK-8 assay. HUVECs cultured with amiodarone of this concentration were set as the experimental groups and those without amiodarone were set as the control group. Apoptosis rate of HUVECs was detected by Annexin V-FITC/P flow cytometry; the protein and mRNA expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase-3, interleukin 10 (IL-10), IL-1β, IL-6, and tumor necrosis factor alpha (TNF-α) were detected using western blotting and real-time fluorescence quantitative polymerase chain reaction, respectively; the reactive oxygen species (ROS) was detected by DCFH-DA fluorescence probe assay; the superoxide dismutase (SOD) activity was detected by water-soluble tetrazolium-1 assay; the reduced glutathione (GSH) content was detected by microplate assay.Results:The viabilities of HUVECs incubated with amiodarone at concentration of 10, 20, 30, and 60 μmol/L for 24 hours were (88.82±2.64)%, (74.96±1.75)%, (64.95±2.10)%, and (18.57±0.65)%, respectively; differences were all significant (all P<0.01) between each experiment group and control group, as well as between each experiment group. Amiodarone at a concentration of 30 μmol/L was used for subsequent experiments. After incubating with 30 μmol/L amiodarone for 6, 12, 24, 36, and 48 hours, the viabilities of HUVECs were (90.19±1.88)%, (82.81±2.51)%, (75.33±1.37)%, (65.76±1.85)%, and (47.01±3.29)%, respectively; differences were all significant (all P<0.01) between each experiment group and control group, as well as between each experiment group. Compared with the control group, the apoptosis rate of cells in the experimental group was significantly higher (48.59% vs. 16.34%, P<0.01), the protein and mRNA expression levels of pro-apoptotic proteins Bax and caspase-3, and pro-inflammatory factors IL-1β, IL-6, and TNF-α were higher (all P<0.01), whereas the protein and mRNA expression levels of anti-apoptotic protein Bcl-2 and anti-inflammatory factor IL-10 were lower ( P<0.05, P<0.01). Conclusions:Amiodarone can cause HUVECs injury, which would be enhanced with the increase of concentration and action time of amiodarone. Amiodarone may cause HUVECs injury by inducing apoptosis, inflammatory response, and oxidative stress.

Objective:To explore the injury effect and its possible mechanism of amiodarone on human umbilical vein endothelial cells (HUVECs).Methods:After 3 generations of cultivation, the HUVECs were seeded in 96-well plates and incubated with amiodarone (0, 10, 20, 30, and 60 μmol/L) for 24 hours. The cell viability was detected using cell counting kit 8 (CCK-8) assay and the relative viability of cells incubated with different concentrations of amiodarone were calculated by taking the cell viability of the 0 μmol/L group as 100%. The concentration of amiodarone at which cell viability was reduced to 70% was selected for subsequent experiments. The effect of amiodarone of this concentration on the activity of HUVECs after action for different time (6, 12, 24, 36, and 48 hours) was detected using the CCK-8 assay. HUVECs cultured with amiodarone of this concentration were set as the experimental groups and those without amiodarone were set as the control group. Apoptosis rate of HUVECs was detected by Annexin V-FITC/P flow cytometry; the protein and mRNA expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase-3, interleukin 10 (IL-10), IL-1β, IL-6, and tumor necrosis factor alpha (TNF-α) were detected using western blotting and real-time fluorescence quantitative polymerase chain reaction, respectively; the reactive oxygen species (ROS) was detected by DCFH-DA fluorescence probe assay; the superoxide dismutase (SOD) activity was detected by water-soluble tetrazolium-1 assay; the reduced glutathione (GSH) content was detected by microplate assay.Results:The viabilities of HUVECs incubated with amiodarone at concentration of 10, 20, 30, and 60 μmol/L for 24 hours were (88.82±2.64)%, (74.96±1.75)%, (64.95±2.10)%, and (18.57±0.65)%, respectively; differences were all significant (all P<0.01) between each experiment group and control group, as well as between each experiment group. Amiodarone at a concentration of 30 μmol/L was used for subsequent experiments. After incubating with 30 μmol/L amiodarone for 6, 12, 24, 36, and 48 hours, the viabilities of HUVECs were (90.19±1.88)%, (82.81±2.51)%, (75.33±1.37)%, (65.76±1.85)%, and (47.01±3.29)%, respectively; differences were all significant (all P<0.01) between each experiment group and control group, as well as between each experiment group. Compared with the control group, the apoptosis rate of cells in the experimental group was significantly higher (48.59% vs. 16.34%, P<0.01), the protein and mRNA expression levels of pro-apoptotic proteins Bax and caspase-3, and pro-inflammatory factors IL-1β, IL-6, and TNF-α were higher (all P<0.01), whereas the protein and mRNA expression levels of anti-apoptotic protein Bcl-2 and anti-inflammatory factor IL-10 were lower ( P<0.05, P<0.01). Conclusions:Amiodarone can cause HUVECs injury, which would be enhanced with the increase of concentration and action time of amiodarone. Amiodarone may cause HUVECs injury by inducing apoptosis, inflammatory response, and oxidative stress.