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.

Background and Aims:Totally implantable venous access ports(TIVAP)are widely used in patients requiring long-term intravenous therapy.Traditional butterfly needle puncture fixation methods have limitations,including low success rates,increased pain,and risk of needle-stick injury.This study aimed to design an adjustable puncture protection kit for butterfly needles and evaluate its clinical utility using a simulated device.Methods:A prospective randomized controlled trial was conducted with 70 patients implanted with upper arm ports in the Hematology Department of Xiangya Hospital,Central South University,from January to December 2024.The patients were divided into a study group and a control group,with 35 cases in each,using a randomized block design.The study group underwent puncture with the simulated adjustable protection kit,while the control group used the traditional finger fixation method.Outcomes compared included first-attempt success rate,vertical puncture rate,pain score,puncture time,and complication rate.Results:The baseline characteristics of the two groups were balanced.The study group had significantly higher first-attempt puncture success rate and vertical puncture rate than the control group(94.3%vs.77.1%;91.4%vs.57.1%,both P<0.05).In the experimental group compared with the control group,pain scores were lower(1.80±1.13 vs.2.94±1.33,P<0.05),and puncture time was shorter[(31.31±9.05)s vs.(41.80±23.97)s,P<0.05].There was no significant difference in the incidence of puncture-related complications between the two groups(2.9%vs.14.3%,P>0.05).Conclusion:The simulated adjustable butterfly needle puncture protection kit effectively improves puncture success,enhances efficiency,reduces patient pain,and demonstrates good clinical safety.This innovative design provides a promising solution for reducing needle-stick injury risks and optimizing port puncture procedures,although larger,multicenter,and long-term studies are warranted.

Aim To investigate the possible mechanism of the myocardial protective effect of Danzhi Jiangtang capsules(DJC)on db/db mice based on NLRP3 in-flammasome-mediated pyroptosis.Methods The db/db mice were randomly divided into the model group,DJC low,medium,and high dose groups,and the met-formin group,and the db/m mice were taken as the blank group.The administration lasted for eightweeks.At the end of drug administration,blood glucose,blood lipids,cardiac enzymes and inflammatory factors were detected in each group of mice.HE and Masson stai-ning was performed to observe the morphology and fi-brosis of myocardial tissue.TUNEL staining was per-formed to detect apoptosis.RT-qPCR was performed to detect the mRNA expression of ANP,BNP and β-MHC,and Western blot was performed to detect the protein expression of NLRP3,ASC,caspase-1,cleaved-caspase-1,GSDMD and GSDMD-NT in myocardial tis-sue.Results DJC could alleviate myocardial patho-logical damage,reduce collagen deposition and apopto-sis,reduce the levels of blood glucose,blood lipid,myo-cardial enzyme and inflammatory factors in db/db mice.DJC could reduce the mRNA expressions of ANP,BNP and β-MHC,and the protein expressions of NLRP3,ASC,caspase-1,cleavedcaspase-1,GSDMD and GSDMD-NT in myocardial tissues.Conclusion DJC attenuates myocardial injury in db/db mice,prob-ably by inhibiting the activation of NLRP3 inflamma-somes,attenuating cardiomyocyte pyroptosis,and amel-iorating the inflammatory state.

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.

Background and Aims:Totally implantable venous access ports(TIVAP)are widely used in patients requiring long-term intravenous therapy.Traditional butterfly needle puncture fixation methods have limitations,including low success rates,increased pain,and risk of needle-stick injury.This study aimed to design an adjustable puncture protection kit for butterfly needles and evaluate its clinical utility using a simulated device.Methods:A prospective randomized controlled trial was conducted with 70 patients implanted with upper arm ports in the Hematology Department of Xiangya Hospital,Central South University,from January to December 2024.The patients were divided into a study group and a control group,with 35 cases in each,using a randomized block design.The study group underwent puncture with the simulated adjustable protection kit,while the control group used the traditional finger fixation method.Outcomes compared included first-attempt success rate,vertical puncture rate,pain score,puncture time,and complication rate.Results:The baseline characteristics of the two groups were balanced.The study group had significantly higher first-attempt puncture success rate and vertical puncture rate than the control group(94.3%vs.77.1%;91.4%vs.57.1%,both P<0.05).In the experimental group compared with the control group,pain scores were lower(1.80±1.13 vs.2.94±1.33,P<0.05),and puncture time was shorter[(31.31±9.05)s vs.(41.80±23.97)s,P<0.05].There was no significant difference in the incidence of puncture-related complications between the two groups(2.9%vs.14.3%,P>0.05).Conclusion:The simulated adjustable butterfly needle puncture protection kit effectively improves puncture success,enhances efficiency,reduces patient pain,and demonstrates good clinical safety.This innovative design provides a promising solution for reducing needle-stick injury risks and optimizing port puncture procedures,although larger,multicenter,and long-term studies are warranted.

Objective To evaluate the efficacy of myopia progression control over a one-year period for defocus in-corporated multiple segments lenses(DIMS),peripheral defocus modifying(PDM)lenes and single vision spectacle lenses(SVL)in a real-world cohort.Methods A total of 188 cases in the DIMS group,116 cases in the PDM group,and 373 ca-ses in the SVL group,all diagnosed with refractive errors and aged 6-16 years,were enrolled in Department of Ophthal-mology,Fujian Provincial Hospital from July 2019 to June 2022.A 1∶1 propensity score matching(PSM)of covariates was used to balance factors between the groups.The changes in spherical equivalent(SE)and axial length(AL)in each patient before and 1 year after wearing glasses were compared without other interventions.The myopia control effects of different groups were compared through the changes in SE and AL.Results After PSM,92 cases were matched between DIMS group and PDM group,163 cases were matched between DIMS group and SVL group,and 99 cases were matched between PDM group and SVL group.The changes in SE and AL in the DIMS group were less than those in the PDM and SVL groups,with statistically significant differences(all P＜0.05).However,there were no statistically significant differences in the changes in SE and AL between the PDM and SVL groups(all P＞0.05).In the age subgroup analysis,in the younger age group(6～＜13)and the older age group(13～16)the changes in SE and AL in the DIMS group were less than those in the PDM group and SVL group,with statistically significant differences(all P＜0.05).In the younger age group,the changes in SE and AL in the PDM group were less than those in the SVL group,with statistically significant differences(all P＜0.05);while in older age group,there were no statistically significant differences in the changes in SE and AL between the PDM and SVL groups(all P＞0.05).Conclusion The myopia control efficacy of DIMS is superior to that of PDM and SVL.In younger children,the myopia control efficacy of PDM is superior to that of SVL.

Aim To investigate the effect of PU.1 in-hibitor DB2313 on lupus nephritis in MRL/lpr mice and its mechanism.Methods Thirty female MRL/lpr mice were randomly divided into the model group,DB2313 group and TACI-Ig group,with 10 mice in each group.Another 10 female BALB/c mice were se-lected as normal control groups.Mice in the DB2313 group received intraperitoneal DB2313 injections every two days,and those in the TACI-Ig group received subcutaneous injections of TACI-Ig every two days.Mice in the control group and model group were intra-gastrically given the same amount of 0.9％NaCl injec-tion every day.Before the drug intervention and for 1 to 5 weeks after the intervention,the urine of mice was collected regularly,the urine protein content was meas-ured,and the renal damage index was evaluated.The histopathological changes of kidney were observed by HE,Masson and PAS staining.The expression levels of immune complex of C3 in kidney tissue were detec-ted by immunohistochemistry.The concentrations of u-rea nitrogen(BUN),serum creatinine(Scr),inter-leukin-6(IL-6),and tumor necrosis factor alpha(TNF-α)in the serum samples were assayed utilizing the respective kits.The expression levels of PU.1 and FLT3 in kidney tissues were determined by immunoflu-orescence technology,and the protein expressions of PU.1,FLT3,PI3K,AKT and phosphorylated AKT(p-AKT)in kidney tissues were detected by Western blot.Results DB2313 treatment significantly allevia-ted the pathological damage of kidney in MRL/lpr mice,and reduced the deposition of C3,kidney injury index and 24-hour urine protein in renal tissue.The results of ELISA showed that DB2313 administration could significantly reduce the serum levels of BUN,Scr,IL-6 and TNF-α in MRL/lpr mice.The results of immunofluorescence and Western blot further showed that DB2313 treatment could significantly down-regu-late the protein expression of PU.1,PI3K and p-AKT,and up-regulate the protein expression of FLT3.Con-clusion DB2313 has an ameliorating effect on lupus nephritis in MRL/lpr mice,and its underlying mecha-nism may involve the inhibition of the transcription fac-tor PU.1-mediated signaling pathway.

Aim To investigate the possible mechanism of the myocardial protective effect of Danzhi Jiangtang capsules(DJC)on db/db mice based on NLRP3 in-flammasome-mediated pyroptosis.Methods The db/db mice were randomly divided into the model group,DJC low,medium,and high dose groups,and the met-formin group,and the db/m mice were taken as the blank group.The administration lasted for eightweeks.At the end of drug administration,blood glucose,blood lipids,cardiac enzymes and inflammatory factors were detected in each group of mice.HE and Masson stai-ning was performed to observe the morphology and fi-brosis of myocardial tissue.TUNEL staining was per-formed to detect apoptosis.RT-qPCR was performed to detect the mRNA expression of ANP,BNP and β-MHC,and Western blot was performed to detect the protein expression of NLRP3,ASC,caspase-1,cleaved-caspase-1,GSDMD and GSDMD-NT in myocardial tis-sue.Results DJC could alleviate myocardial patho-logical damage,reduce collagen deposition and apopto-sis,reduce the levels of blood glucose,blood lipid,myo-cardial enzyme and inflammatory factors in db/db mice.DJC could reduce the mRNA expressions of ANP,BNP and β-MHC,and the protein expressions of NLRP3,ASC,caspase-1,cleavedcaspase-1,GSDMD and GSDMD-NT in myocardial tissues.Conclusion DJC attenuates myocardial injury in db/db mice,prob-ably by inhibiting the activation of NLRP3 inflamma-somes,attenuating cardiomyocyte pyroptosis,and amel-iorating the inflammatory state.

Objective:To investigate the changes of the epidemiology and clinical characteristics of human metapneumovirus (hMPV) among children with acute lower respiratory tract infection (ALRTI) before and after the discontinuation of non-pharmaceutical interventions (NPI) during coronavirus disease 2019 epidemic.Methods:This was a retrospective cohort study. Children hospitalized at The Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University between January 2021 and December 2023, who were diagnosed with ALRTI by nasopharyngeal secretion testing for respiratory pathogens nucleic acid were enrolled. Clinical and laboratory data were collected. Children admitted between January 1, 2021 and January 7, 2023 were classified as the pre-NPI withdrawal group (abbreviated as pre-withdrawal group), while those admitted from January 8, 2023 afterward were classified as the post-NPI withdrawal group (abbreviated as post-withdrawal group). Nasopharyngeal secretions from the enrolled children were tested for 13 respiratory pathogens using polymerase chain reaction-capillary electrophoresis fragment analysis, and bacterial cultures were also performed. Statistical analyses were performed using the Mann-Whitney U test or chi-square test. Results:A total of 30 855 ALRTI cases were enrolled, with 1 679 of hMPV-positive. In the pre-withdrawal group, there were 861 cases with an age of onset of 2.0(1.0, 3.0) years, and the highest proportion was in the 1 to <3 years age group, accounting for 35.3%(304/861). In the post-withdrawal group, there were 818 cases with an age of onset of 3.0(2.0, 4.0) years, and the highest proportion was in the 3 to <5 years age group, accounting for 39.2%(321/818).The age of onset in the post-withdrawal group was significantly older than that in the pre-withdrawal group ( Z=7.69, P<0.001) .The hMPV detection rate was higher in the pre-withdrawal group than that in post-withdrawal group (5.75%(861/14 984) vs 5.15%(818/15 871); χ2=5.25, P=0.022). In the pre-withdrawal group, the epidemic peaks occurred in winter and spring, with the highest rates in January 2022(25.2%(224/890)) and March 2022 (21.6%(186/860)). In the post-withdrawal group, the epidemic peak shifted to spring and summer, and the detection rate became increased since April 2023(10.8%(136/1 258)). The post-withdrawal group showed lower rates of wheezing, shortness of breath, cyanosis, respiratory support, severe pneumonia, intensive care unit admission, and shorter hospital stays compared to the pre-withdrawal group ( χ2=69.09, 31.63, 12.97, 57.96, 55.73, 5.48 and Z=7.11, respectively, all P< 0.05).In the pre-withdrawal group, 412 cases (47.9%(412/861)) had other pathogens detected, compared to 445 cases (54.4%(445/181)) in the post-withdrawal group, indicating a significantly higher rate of co-infections in the post-withdrawal group ( χ2=7.20, P<0.05). The most commonly detected pathogens in both groups were Mycoplasma pneumoniae (MP), rhinovirus, and Streptococcus pneumoniae. However, the post-withdrawal group showed significantly higher detection rates of MP and influenza virus, but lower bacterial detection rates compared to the pre-withdrawal group ( χ2=39.41, 9.70, 5.63, respectively, all P<0.05). The detection rate of Haemophilus influenzae was 2.1%(17/818) in the post-withdrawal group which lower than that (6.7%(58/861)) in the pre-withdrawal group, and the difference was statistically significant ( χ2=21.32, P<0.001). Conclusions:In 2023, following the withdrawal of NPI, the epidemic peak of hMPV in Wenzhou area is delayed to spring and summer. The age of children with hMPV-associated ALRTI increases, with the majority being 3 to <5 years old. The overall severity of the disease decreases. However, the detection of mixed pathogens increases, with MP being the most common, while bacterial detection decreases.

This study explored the potential therapeutic targets and mechanisms of Danggui Shaoyao San(DSS) in the prevention and treatment of Alzheimer's disease(AD) through transcriptomics and metabolomics, combined with animal experiments. Fifty male C57BL/6J mice, aged seven weeks, were randomly divided into the following five groups: control, model, positive drug, low-dose DSS, and high-dose DSS groups. After the intervention, the Morris water maze was used to assess learning and memory abilities of mice, and Nissl staining and hematoxylin-eosin(HE) staining were performed to observe pathological changes in the hippocampal tissue. Transcriptomics and metabolomics were employed to sequence brain tissue and identify differential metabolites, analyzing key genes and metabolites related to disease progression. Reverse transcription-quantitative polymerase chain reaction(RT-qPCR) was employed to validate the expression of key genes. The Morris water maze results indicated that DSS significantly improved learning and cognitive function in scopolamine(SCOP)-induced model mice, with the high-dose DSS group showing the best results. Pathological staining showed that DSS effectively reduced hippocampal neuronal damage, increased Nissl body numbers, and reduced nuclear pyknosis and neuronal loss. Transcriptomics identified seven key genes, including neurexin 1(Nrxn1) and sodium voltage-gated channel α subunit 1(Scn1a), and metabolomics revealed 113 differential metabolites, all of which were closely associated with synaptic function, oxidative stress, and metabolic regulation. RT-qPCR experiments confirmed that the expression of these seven key genes was consistent with the transcriptomics results. This study suggests that DSS significantly improves learning and memory in SCOP model mice and alleviates hippocampal neuronal pathological damage. The mechanisms likely involve the modulation of synaptic function, reduction of oxidative stress, and metabolic balance, with these seven key genes serving as important targets for DSS in the treatment of AD.
Animals ; Alzheimer Disease/genetics* ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Mice, Inbred C57BL ; Metabolomics ; Transcriptome/drug effects* ; Maze Learning/drug effects* ; Hippocampus/metabolism* ; Humans ; Disease Models, Animal ; Memory/drug effects*