OBJECTIVES: To investigate the clinical features of children with STAT gene mutations, and to explore corresponding immunotherapy strategies. METHODS: A retrospective analysis was performed for the clinical data of 10 children with STAT gene mutations who were admitted to the Department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University, from October 2015 to October 2024. Exploratory immunotherapy was implemented in some refractory cases, and the changes in symptoms, imaging manifestations, and cytokine levels were assessed after treatment. RESULTS: For the 10 children, the main clinical manifestations were recurrent rash since birth (7/10), cough (8/10), wheezing (5/10), expectoration (4/10), and purulent nasal discharge (4/10). Genotyping results showed that there was one child with heterozygous loss-of-function (LOF) mutation in the STAT1 gene, four children with heterozygous LOF mutation in the STAT3 gene, and five children with heterozygous gain-of-function (GOF) mutation in the STAT3 gene. Two children with LOF mutation in the STAT3 gene showed decreased interleukin-6 levels and improved clinical symptoms and imaging findings after omalizumab treatment. Three children with GOF mutation in the STAT3 gene achieved effective disease control after treatment with methylprednisolone (0.5 mg/kg per day). Two children with GOF mutation in the STAT3 gene received treatment with JAK inhibitor and then showed some improvement in symptoms. CONCLUSIONS STAT gene mutation screening should be considered for children with recurrent rash and purulent respiratory tract infections. Targeted immunotherapy may improve prognosis in patients with no response to conventional treatment.
Humans ; Male ; Immunotherapy ; Female ; Child, Preschool ; Child ; Gain of Function Mutation ; Retrospective Studies ; Infant ; Loss of Function Mutation ; STAT Transcription Factors/genetics*

The innate immune sensor NLRP3 inflammasome overactivation is involved in the pathogenesis of ulcerative colitis. PGAM5 is a mitochondrial phosphatase involved in NLRP3 inflammasome activation in macrophages. However, the role of PGAM5 in ulcerative colitis and the mechanisms underlying PGAM5 regulating NLRP3 activity remain unknown. Here, we show that PGAM5 deficiency ameliorates dextran sodium sulfate (DSS)-induced colitis in mice via suppressing NLRP3 inflammasome activation. By combining APEX2-based proximity labeling focused on PGAM5 with quantitative proteomics, we identify NEK7 as the new binding partner of PGAM5 to promote NLRP3 inflammasome assembly and activation in a PGAM5 phosphatase activity-independent manner upon inflammasome induction. Interfering with PGAM5-NEK7 interaction by punicalagin inhibits the activation of the NLRP3 inflammasome in macrophages and ameliorates DSS-induced colitis in mice. Altogether, our data demonstrate the PGAM5-NEK7 interaction in macrophages for NLRP3 inflammasome activation and further provide a promising therapeutic strategy for ulcerative colitis by blocking the PGAM5-NEK7 interaction.

AIM To optimize the cellulase-assisted ultrasound extraction process for total flavonoids from Plumbago zeylanica L.,and to evaluate their anti-oxidant activity.METHODS With extraction time,liquid-solid ratio,cellulase addition amount,extraction temperature and ultrasonic power as influencing factors,extraction rate of total flavonoids as an evaluation index,the extraction process was optimized by response surface method on the basis of single factor test.Subsequently,The scavenging rates of extract on DPPH,ABTS and OH free radicals were determined.RESULTS The optimal conditions were determined to be 34∶1 for liquid-solid ratio,3％for cellulase addition amount,51 ℃ for extraction temperature,38 min for extraction time,and 400 W for ultrasonic power,the extraction rate of total flavonoids was(33.411±0.97)％.The IC50 values of three free radicals were 0.13,0.042,3.29 mg/mL,respectively.CONCLUSION This reasonable and reliable method can be used for the cellulase-assisted ultrasound extraction of total flavonoids from P.zeylanica with strong anti-oxidant activity.

Objective To explore the mechanism of action of mitochondrial DNA(mtDNA)of alveolar epithelial type Ⅱ cells(AECⅡ)in silicotic fibrosis.Methods This study was divided into the animal experiment and cell experiment.Animal experiment:twenty-eight C57BL/6 mice were randomly divided into the saline group(n=14)and the silicosis model group(n=14),which were given 50 μL of sterile saline and 50 μL of 50 mg/mL of SiO2 suspension by tracheal perfusion respectively.HE staining and Masson staining were performed to observe the histopathological changes of mouse lung tissues after sampling with the exposure of 28 days and 56 days,and Western blot was used to detect the expression levels of functional proteins of pulmonary surfactant protein-D(SP-D),and Krebs von den Lungen-6(KL-6),and fibrotic protein of transforming growth factor-β1(TGF-β1)in mouse lung tissues.Cell experiment:AECⅡ cell line of A549 was exposed to different concentrations of SiO2 suspension.CCK-8 was used to detect cell viability,DAPI staining was used to observe cell apoptosis,Western blot was used to detect related protein expression,JC-1 was used to detect the changes of mitochondrial membrane potential,and RT-qPCR was used to detect mtDNA levels in whole cells,mitochondria,cytoplasm and supernatant.The expression levels of related proteins in AECⅡ were detected by Western blot after cleared mtDNA by DnaseⅠ.Results Histopathological examination of lung tissues showed that SiO2 exposure led to damage in mice lung tissues,with infiltration of inflammatory cell,silica nodules,and a significant increase of collagen fibers.After SiO2 exposure,the protein expressions of SP-D,KL-6 and TGF-β1 in lung tissues were up-regulated(P<0.05).DAPI staining and CCK-8 results showed a reduced activity and impaired apoptosis of AECⅡ after treated with 400 and 800 μg/mL SiO2.JC-1 assay results demonstrated that exposure to 800 μg/mL SiO2 caused a decrease of mitochondrial membrane potential in AECⅡ(P<0.05).RT-qPCR results showed that 800 μg/mL SiO2 exposure led to increased mtDNA levels in the cytoplasm and supernatant of AECⅡ(P<0.05).Western blot results showed that 800 μg/mL SiO2 exposure caused an increase in the levels of SP-D,KL-6,and TGF-β1 proteins,while which were ameliorated by Dnase Ⅰ treatment.Conclusion SiO2 exposure results in the escape and release of AECⅡ mtDNA and dysfunction of AECⅡ,thereby leading to the development of pulmonary fibrosis.

ObjectiveTo investigate the spontaneous premature cardiac aging in SHJH^hr mice. MethodsA comparative study was conducted between SHJH^hr mice (SHJH^hr group) and wild-type ICR mice (WT group) at different ages (10 and 24 weeks). Cardiac function was analyzed using a small animal in vivo ultrasound imaging system. After euthanasia, organs were collected and weighed to assess the extent of cardiac atrophy. Cardiac pathological damage was observed using hematoxylin-eosin (HE) staining. Cardiac fibrosis was analyzed using Masson staining. Myocardial cell area was analyzed after wheat germ agglutinin (WGA) staining. The activities of oxidative damage indicators in myocardial tissue, including superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT), as well as the content of 8-hydroxy-2'-deoxyguanosine (8-OHdG), were measured using enzyme-linked immunosorbent assay. Real-time fluorescence quantitative PCR was used to measure the mRNA expression levels of factors associated with inflammation, fibrosis, and oxidative stress. Colorimetric assay was used to measure malondialdehyde (MDA) levels. ResultsCompared to WT group mice of the same age, 10-week-old mice in the SHJH^hr group showed no significant differences in stroke volume (SV), ejection fraction (EF), fractional shortening (FS), or heart and lung weights. However, at 24 weeks of age, mice in the SHJH^hr group had significantly lower SV, EF, and FS values compared to mice of the same age in the WT group (P<0.05), with no significant change in lung weight but a significant reduction in heart weight (P<0.05). Histological analysis of heart tissue from 24-week-old mice revealed no significant difference in cardiac fibrosis levels between SHJH^hr and WT groups, but WGA staining showed a significant reduction in myocardial cell area in mice in the SHJH^hr group (P<0.05). PCR analysis revealed a significant downregulation of mRNA levels of oxidative stress factors Sod2, Gpx1, and Cat genes (P<0.05). Biochemical assays indicated significantly reduced activities of oxidative damage-related enzymes SOD, GPX, and CAT in myocardial tissue (P<0.05), while the levels of oxidative damage markers 8-OHdG and MDA significantly increased (P<0.05). ConclusionMice in the SHJH^hr group exhibit premature cardiac aging, which may be associated with oxidative stress damage in myocardial tissue.

Objective: To explore the clinical safety and efficacy of a single-position robot-assisted radical nephroureterectomy (RRUN) in the treatment of upper tract urothelial carcinoma (UTUC). Methods: A retrospective study was conducted on 136 UTUC patients who underwent RRUN (n=35) and laparoscopic radical nephroureterectomy (LRUN，n=101) in our hospital during Dec.2020 and Aug.2023.The perioperative and safety indicators of the two groups were compared.The intravesical recurrence-free survival (IVRFS)，recurrence-free survival (RFS)，and overall survival (OS) of the two groups were compared using Kaplan-Meier method. Results: There were no significant differences in the baseline data between the two groups (P>0.05).All surgeries were successfully completed without conversion to open surgery.RRUN demonstrated superior perioperative outcomes compared to LRUN in overall postoperative complication rate [37.1%(13/35) vs. 56.4%(57/101)]，postoperative hospital stay [6(5,7) days vs. 7(6,8) days]，and catheter indwelling time [3(2，4) days vs. 4(3，5) days]，with statistically significant differences (P<0.05).Safety indicators of both surgical approaches were similar (P>0.05).Survival analysis showed no significant difference in oncological outcomes between the two groups [IVRFS (1 year：92.1%，2 years：85.2%)，RFS (1 year:82.4%，2 years:74.9%)，OS (1 year:90.6%，2 years:84.2%)] (P>0.05). Conclusion: Compared with retroperitoneal LRUN，single-position RRUN for UTUC demonstrates comparable safety and oncological efficacy，while offering significant advantages in perioperative outcomes such as reducing postoperative complication rate and shortening hospital stay.

Objective To investigate the effectiveness of new-generation snapshot freeze(NG SSF)technology for improving image quality of coronary CT angiography(CCTA)in patients with different heart rate(HR).Methods CCTA data of 164 patients obtained with a 256-row CT scanner and voltages of 80,100 and 120 kV,respectively,in one cardiac cycle were retrospectively analyzed.Smart electrocardiogram(ECG)gating technology was used to calculate patient's HR and choose automatic exposure phase.For these with HR≤65 beats/min(low HR group),the exposure time window was set between 70%—80%R-R interval,for 65 beats/min＜HR≤85 beats/min(medium HR group)was set between 40%—80%R-R interval,while for those HR＞85 beats/min(high HR group)was set between 40%—60%R-R interval.Standard reconstruction(STD),the first-generation snapshot freeze(SSF1)and NG SSF were performed to reconstruct CCTA images,respectively.Subjective scoring of each segment of right coronary artery(RCA),left anterior descending(LAD)and left circumflex artery(LCX)shown on CCTA were performed based on Likert scale.Results In low HR group,the scores of middle and distal segments of LAD and all segments of RCA and LCX on NG SSF CCTA were significantly higher than on STD,and of middle segment of RCA and distal segment of LAD were both higher than on SSF1(all P＜0.05).In medium HR group,the scores of all branches on NG SSF CCTA were significantly higher than on STD or SSF1 CCTA(all P＜0.05).In high HR group,the scores of coronary branches showed on NG SSF CCTA were all significantly higher than on STD CCTA,while of the proximal and distal segments of RCA,middle and distal segments of LAD and all segments of LCX were significantly higher than on SSF1 CCTA(all P＜0.05).Conclusion NG SSF could effectively enhance image quality of prospective ECG-gated CCTA in patients with varying HR.

Objective To explore the mechanism of action of mitochondrial DNA(mtDNA)of alveolar epithelial type Ⅱ cells(AECⅡ)in silicotic fibrosis.Methods This study was divided into the animal experiment and cell experiment.Animal experiment:twenty-eight C57BL/6 mice were randomly divided into the saline group(n=14)and the silicosis model group(n=14),which were given 50 μL of sterile saline and 50 μL of 50 mg/mL of SiO2 suspension by tracheal perfusion respectively.HE staining and Masson staining were performed to observe the histopathological changes of mouse lung tissues after sampling with the exposure of 28 days and 56 days,and Western blot was used to detect the expression levels of functional proteins of pulmonary surfactant protein-D(SP-D),and Krebs von den Lungen-6(KL-6),and fibrotic protein of transforming growth factor-β1(TGF-β1)in mouse lung tissues.Cell experiment:AECⅡ cell line of A549 was exposed to different concentrations of SiO2 suspension.CCK-8 was used to detect cell viability,DAPI staining was used to observe cell apoptosis,Western blot was used to detect related protein expression,JC-1 was used to detect the changes of mitochondrial membrane potential,and RT-qPCR was used to detect mtDNA levels in whole cells,mitochondria,cytoplasm and supernatant.The expression levels of related proteins in AECⅡ were detected by Western blot after cleared mtDNA by DnaseⅠ.Results Histopathological examination of lung tissues showed that SiO2 exposure led to damage in mice lung tissues,with infiltration of inflammatory cell,silica nodules,and a significant increase of collagen fibers.After SiO2 exposure,the protein expressions of SP-D,KL-6 and TGF-β1 in lung tissues were up-regulated(P<0.05).DAPI staining and CCK-8 results showed a reduced activity and impaired apoptosis of AECⅡ after treated with 400 and 800 μg/mL SiO2.JC-1 assay results demonstrated that exposure to 800 μg/mL SiO2 caused a decrease of mitochondrial membrane potential in AECⅡ(P<0.05).RT-qPCR results showed that 800 μg/mL SiO2 exposure led to increased mtDNA levels in the cytoplasm and supernatant of AECⅡ(P<0.05).Western blot results showed that 800 μg/mL SiO2 exposure caused an increase in the levels of SP-D,KL-6,and TGF-β1 proteins,while which were ameliorated by Dnase Ⅰ treatment.Conclusion SiO2 exposure results in the escape and release of AECⅡ mtDNA and dysfunction of AECⅡ,thereby leading to the development of pulmonary fibrosis.

Objective To investigate the effectiveness of new-generation snapshot freeze(NG SSF)technology for improving image quality of coronary CT angiography(CCTA)in patients with different heart rate(HR).Methods CCTA data of 164 patients obtained with a 256-row CT scanner and voltages of 80,100 and 120 kV,respectively,in one cardiac cycle were retrospectively analyzed.Smart electrocardiogram(ECG)gating technology was used to calculate patient's HR and choose automatic exposure phase.For these with HR≤65 beats/min(low HR group),the exposure time window was set between 70%—80%R-R interval,for 65 beats/min＜HR≤85 beats/min(medium HR group)was set between 40%—80%R-R interval,while for those HR＞85 beats/min(high HR group)was set between 40%—60%R-R interval.Standard reconstruction(STD),the first-generation snapshot freeze(SSF1)and NG SSF were performed to reconstruct CCTA images,respectively.Subjective scoring of each segment of right coronary artery(RCA),left anterior descending(LAD)and left circumflex artery(LCX)shown on CCTA were performed based on Likert scale.Results In low HR group,the scores of middle and distal segments of LAD and all segments of RCA and LCX on NG SSF CCTA were significantly higher than on STD,and of middle segment of RCA and distal segment of LAD were both higher than on SSF1(all P＜0.05).In medium HR group,the scores of all branches on NG SSF CCTA were significantly higher than on STD or SSF1 CCTA(all P＜0.05).In high HR group,the scores of coronary branches showed on NG SSF CCTA were all significantly higher than on STD CCTA,while of the proximal and distal segments of RCA,middle and distal segments of LAD and all segments of LCX were significantly higher than on SSF1 CCTA(all P＜0.05).Conclusion NG SSF could effectively enhance image quality of prospective ECG-gated CCTA in patients with varying HR.

AIM To optimize the cellulase-assisted ultrasound extraction process for total flavonoids from Plumbago zeylanica L.,and to evaluate their anti-oxidant activity.METHODS With extraction time,liquid-solid ratio,cellulase addition amount,extraction temperature and ultrasonic power as influencing factors,extraction rate of total flavonoids as an evaluation index,the extraction process was optimized by response surface method on the basis of single factor test.Subsequently,The scavenging rates of extract on DPPH,ABTS and OH free radicals were determined.RESULTS The optimal conditions were determined to be 34∶1 for liquid-solid ratio,3％for cellulase addition amount,51 ℃ for extraction temperature,38 min for extraction time,and 400 W for ultrasonic power,the extraction rate of total flavonoids was(33.411±0.97)％.The IC50 values of three free radicals were 0.13,0.042,3.29 mg/mL,respectively.CONCLUSION This reasonable and reliable method can be used for the cellulase-assisted ultrasound extraction of total flavonoids from P.zeylanica with strong anti-oxidant activity.