Atopic dermatitis （AD） is an atopic disease with a complex etiology and pathogenesis resulting from the interaction of multiple factors. The NOD-like receptor pyrin domain-containing 3 （NLRP3） inflammasome is an important component of innate immunity and is involved in the onset and progression of AD， encompassing multiple processes such as inflammation， pyroptosis， and autophagy. Traditional Chinese medicine （TCM） has shown significant clinical efficacy in the treatment of AD and also offers advantages including flexible compatibility， multi-target effects， and low drug resistance. A large number of studies have shown that single Chinese medicinal components and compound prescriptions can treat atopic diseases by modulating the NLRP3 inflammasome. This article elaborates on the activation of the NLRP3 inflammasome and its influence on the pathogenesis and progression of AD， and summarizes recent studies on the mechanisms by which active constituents， extracts， and compound formulations of Chinese medicine treat AD through regulation of the NLRP3 inflammasome and related signaling pathways， with the aim of providing a reference for the clinical treatment of AD and the development of TCM.

BackgroundCurrently， the incidence of depression among teenagers is on the rise， and the related academic problems are becoming increasingly serious. Short video addiction has a negative impact on teenagers' emotional issues and academic achievements. However， few studies have explored the relationship between this addiction and the learning burnout of teenagers with depression， and even fewer have focused on the role paths of impulsivity and coping disposition in this process. ObjectiveTo explore the relationship between short video addiction and learning burnout in adolescent patients with depression， as well as the pathway of impulsivity and coping disposition， so as to provide references for the intervention of learning burnout in adolescent patients with depression. MethodsA total of 191 adolescent patients who were hospitalized at The Affiliated Brain Hospital of Guangzhou Medical University from December 2024 to April 2025 and met the diagnostic criteria for depression according to the International Classification of Diseases， tenth edition （ICD-10） were selected consecutively. The Short Video Addiction Measurement Scale （SVAMS）， the Brief Barratt Impulsiveness Scale （BBIS）， the Simplified Coping Style Questionnaire （SCSQ）， and the Adolescent Student Burnout Inventory （ASBI） were used for assessment. Pearson correlation analysis was employed to examine the correlations of the scores of each scale. Model 6 of the SPSS macro Process 4.2 was employed to analyze the chained mediation pathway of impulsivity and coping disposition between short video addiction and learning burnout. ResultsA total of 173 cases （90.58%） of adolescent patients with depression completed the valid questionnaire survey. Correlation analysis showed that SCSQ coping disposition score was negatively correlated with the SVAMS score， the BBIS score， and the ASBI score （r=-0.282， -0.341， -0.431， P<0.01）， the SVAMS score was positively correlated with the BBIS score and the ASBI score （r=0.339， 0.262， P<0.01）， and the BBIS score was positively correlated with the ASBI score （r=0.486， P<0.01）. The pathway analysis showed that the direct effect of short video addiction on learning burnout was not statistically significant， but the total effect and the indirect effect were statistically significant. The effect values were 0.275 （95% CI： 0.207–0.343） and 0.193 （95% CI： 0.143–0.246）， respectively， with the indirect effect accounting for 70.18%. Impulsivity and coping disposition both played independent mediating roles between short video addiction and learning burnout， with effect values of 0.122 （95% CI： 0.090–0.156） and 0.054 （95% CI： 0.032–0.079）， accounting for 44.36% and 19.64% of the total effect， respectively. The chained mediation effect of impulsivity and coping disposition was significant， with an effect value of 0.017 （95% CI： 0.011–0.026）， accounting for 6.18% of the total effect. ConclusionAlthough short video addiction does not directly affect learning burnout in adolescent patients with depression， it may indirectly influence learning burnout through independent and chain paths of impulsivity and coping disposition. ［Funded by Guangzhou Key Clinical Specialty （Clinical Medical Research Institute）］

Objective To generate a dense granule protein 3 (GRA3) gene-deficient mutant of the Toxoplasma gondii ME49 strain and to test the virulence of the mutant. Methods Gene-deficient parasites were generated with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. Guide RNA (gRNA) was designed using the E-CRISPR software, and mutated on the pSAG1::Cas9-U6::sgUPRT plasmid using the Q5 site-directed mutagenesis kit to generate the pSAG1::Cas9-U6::sgGRA3 plasmid. A GRA3 donor plasmid containing GRA3 gene upstream sequences, pyrimethamine resistant gene dihydrofolate reductase-thymidylate synthase (DHFR-TS) and GRA3 gene downstream sequence was generated, and GRA3 donor DNA was amplified using PCR assay. The pSAG1::Cas9-U6::sgGRA3 plasmid and GRA3 donor DNA were electroporated into tachyzoites of the wild-type T. gondii ME49 strain. Then, parasite suspensions were inoculated into human foreskin fibroblast (HFF) cells and screened with pyrimethamine to yield pyrimethamine-resistant parasites for monoclonal screening. The GRA3 gene deficient monoclonal strain (ME49Δgra3) of T. gondii was identified using PCR and Western blotting assays, and the expression of GRA3 protein was determined in the T. gondii ME49Δgra3 strain using Western blotting. Subsequently, 1 000 freshly lysed tachyzoites of T. gondii ME49 and ME49Δgra3 strains were transferred to 12-well plates seeded with HFF cells, and incubated at 37 °C containing 5% CO₂ for 7 days, and the number of plaques was counted by staining with crystal violet solutions. HFF cells infected with tachyzoites of T. gondii ME49 and ME49Δgra3 strains were stained using Giemsa solutions, and the numbers of cells containing 1, 2, 4, and > 4 T. gondii parasitophorous vacuoles were counted. In addition, the survival rates of C57BL/6 mice infected with T. gondii ME49 and ME49Δgra3 strains were compared 35 days post-infection. Results PCR assay revealed successful amplification of both the upstream and downstream homologous arm bands of the DHFR-TS gene in the T. gondii ME49Δgra3 strain, and no corresponding bands were amplified in the ME49 strain. The GRA3 band was amplified in the ME49 strain, and the DHFR-TS band, rather than GRA3 band, was amplified in the ME49Δgra3 strain. Western blotting determined absence of GRA3 protein expression in the ME49Δgra3 strain. Crystal violet staining showed that the T. gondii ME49 strain produced more plaques than the ME49Δgra3 strain [(352.67 ± 26.39) plaques vs. (235.00 ± 26.29) plaques; t = 5.472, P < 0.01], and Giemsa staining revealed that the proportion of T. gondii parasitophorous vacuoles containing at least four T. gondii tachyzoites was higher in HFF cells infected with the ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(75.67 ± 2.52)% vs. (59.67 ± 2.31)%; t = 8.113, P < 0.01], and the proportion of T. gondii parasitophorous vacuoles containing at least 1 or 2 T. gondii tachyzoites was higher in HFF cells infected with the T. gondii ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(24.33 ± 2.52)% vs. (40.33 ± 2.31)%; t = −8.113, P < 0.01]. In addition, mice infected with the T. gondii ME49 and ME49Δgra3 strains started to die 8 and 9 days post-infection, and the 35-day mortality rates of mice infected with T. gondii ME49 and ME49Δgra3 strains were 10.00% and 70.00% post-infection (χ² = 6.762, P < 0.01). Conclusions The T. gondii ME49Δgra3 strain has been successfully generated, and GRA3 protein may increase the virulence of the T. gondii ME49 strain.

AIM:This study aims to investigate the role and mechanism of proprotein convertase subtilisin/kexin type 9(PCSK9)in ovarian cancer.METHODS:We compared the expression levels of PCSK9 between ovarian cancer specimens and their corresponding adjacent non-cancerous tissues,while also assessing its expression in various ovarian cancer cell lines.Using a shRNA strategy,we reduced the expression of PCSK9 in ovarian cancer cell lines cul-tured in vitro,with confirmation via Western blot.The effects of PCSK9 downregulation on the proliferation,migration,and invasion of ovarian cancer cells were evaluated through EdU,colony formation,and Transwell assays.Additionally,we analyzed the impact of PCSK9 down-regulation on the MAPK/ERK signaling pathway using Western blot analysis.RE-SULTS:PCSK9 was significantly upregulated in ovarian cancer tissues and cell lines(P＜0.01).Downregulation of PC-SK9 resulted in a significant decrease in cell proliferation,migration,and invasion(P＜0.01).Western blot analysis dem-onstrated that PCSK9 knockdown led to reduced expression levels of key molecules within the MAPK/ERK signaling path-way(P＜0.01).CONCLUSION:PCSK9 promotes the proliferation and invasion of ovarian cancer cells by activating MAPK/ERK signaling pathway.

Objective:To investigate the additional value of 99Tc m-hydrazinonicotinamide (HYNIC)-Tyr3-octreotide (TOC) SPECT/CT imaging in the diagnosis and treatment of gastroenteropancreatic neuroendocrine tumors (GEP-NETs). Methods:A total of 54 patients (28 males and 26 females, age: (52.6±11.7) years) who underwent enhanced CT (MR) and 99Tc m-HYNIC-TOC SPECT/CT in People′s Hospital of Zhengzhou University between December 2017 and June 2023 were analyzed retrospectively. Surgical pathology or biopsy was the gold standard of patients′ diagnosis (primary tumors), and comprehensive evaluation based on pathology, imaging and follow-up results was used as the diagnostic criteria of lesions. McNemar χ2 test was used to compare the diagnostic efficacy of different imaging methods. Results:Pathological results showed that 43 of the 54 patients were with GEP-NETs and 11 were with non-neuroendocrine tumors (NETs). The sensitivities of enhanced CT and enhanced MR in the diagnosis of patients with GEP-NETs were 65.1%(28/43) and 60.0%(15/25) respectively, which increased to 93.0%(40/43) and 92.0%(23/25) with the addition of 99Tc m-HYNIC-TOC imaging ( χ2 values: 8.64, 4.90, P values: 0.002, 0.021). There were 22 and 15 patients showing atypical enhancement on enhanced CT and enhanced MR respectively. The sensitivities of these two methods for GEP-NETs in patients with atypical enhancement were 54.5%(12/22) and 8/15 respectively, which increased to 95.5%(21/22) and 14/15 with the addition of 99Tc m-HYNIC-TOC imaging ( χ2 values: 5.82, 4.17, P values: 0.012, 0.031). Compared with enhanced CT, the detection rates of liver and bone metastatic lesions were improved significantly from 90.8%(158/174) and 55.2%(32/58) to 96.6%(168/174) and 87.9%(51/58) with the addition of 99Tc m-HYNIC-TOC imaging ( χ2 values: 5.79, 9.82, P values: 0.013, 0.001). Compared with enhanced MR, the detection rate of bone metastases was improved significantly from 56.0%(14/25) to 88.0%(22/25) with the addition of 99Tc m-HYNIC-TOC imaging ( χ2=4.08, P=0.039). After 99Tc m-HYNIC-TOC imaging, stages were changed in 7.0%(3/43) of patients and a greater number or extent of metastases were detected in 11.6%(5/43) of patients. 99Tc m-HYNIC-TOC imaging detected additional recurrent or metastatic lesions in 40.0%(8/20) of patients during follow-up compared to enhanced CT. Conclusion:99Tc m-HYNIC-TOC imaging can provide an added value for diagnosing GEP-NETs with atypically enhanced CT(MR), and in the detection of liver metastasis and early bone metastasis, thus helping the optimization of clinical treatment strategies.

Objective:To investigate the additional value of 99Tc m-hydrazinonicotinamide (HYNIC)-Tyr3-octreotide (TOC) SPECT/CT imaging in the diagnosis and treatment of gastroenteropancreatic neuroendocrine tumors (GEP-NETs). Methods:A total of 54 patients (28 males and 26 females, age: (52.6±11.7) years) who underwent enhanced CT (MR) and 99Tc m-HYNIC-TOC SPECT/CT in People′s Hospital of Zhengzhou University between December 2017 and June 2023 were analyzed retrospectively. Surgical pathology or biopsy was the gold standard of patients′ diagnosis (primary tumors), and comprehensive evaluation based on pathology, imaging and follow-up results was used as the diagnostic criteria of lesions. McNemar χ2 test was used to compare the diagnostic efficacy of different imaging methods. Results:Pathological results showed that 43 of the 54 patients were with GEP-NETs and 11 were with non-neuroendocrine tumors (NETs). The sensitivities of enhanced CT and enhanced MR in the diagnosis of patients with GEP-NETs were 65.1%(28/43) and 60.0%(15/25) respectively, which increased to 93.0%(40/43) and 92.0%(23/25) with the addition of 99Tc m-HYNIC-TOC imaging ( χ2 values: 8.64, 4.90, P values: 0.002, 0.021). There were 22 and 15 patients showing atypical enhancement on enhanced CT and enhanced MR respectively. The sensitivities of these two methods for GEP-NETs in patients with atypical enhancement were 54.5%(12/22) and 8/15 respectively, which increased to 95.5%(21/22) and 14/15 with the addition of 99Tc m-HYNIC-TOC imaging ( χ2 values: 5.82, 4.17, P values: 0.012, 0.031). Compared with enhanced CT, the detection rates of liver and bone metastatic lesions were improved significantly from 90.8%(158/174) and 55.2%(32/58) to 96.6%(168/174) and 87.9%(51/58) with the addition of 99Tc m-HYNIC-TOC imaging ( χ2 values: 5.79, 9.82, P values: 0.013, 0.001). Compared with enhanced MR, the detection rate of bone metastases was improved significantly from 56.0%(14/25) to 88.0%(22/25) with the addition of 99Tc m-HYNIC-TOC imaging ( χ2=4.08, P=0.039). After 99Tc m-HYNIC-TOC imaging, stages were changed in 7.0%(3/43) of patients and a greater number or extent of metastases were detected in 11.6%(5/43) of patients. 99Tc m-HYNIC-TOC imaging detected additional recurrent or metastatic lesions in 40.0%(8/20) of patients during follow-up compared to enhanced CT. Conclusion:99Tc m-HYNIC-TOC imaging can provide an added value for diagnosing GEP-NETs with atypically enhanced CT(MR), and in the detection of liver metastasis and early bone metastasis, thus helping the optimization of clinical treatment strategies.

Booster vaccinations are highly recommended in combating the SARS-CoV-2 Omicron variant and its subvariants. However, the optimal booster vaccination strategies and related immune mechanisms with different prior vaccinations are under-revealed. In this study, we systematically evaluated the immune responses in mice and hamsters with different prime-boost regimens before their protective efficacies against Omicron were detected. We found that boosting with Ad5-nCoV, S_WT-2P or S_Omicron-6P induced significantly higher levels of neutralization activities against Omicron variants than CoronaVac and ZF2001 by eliciting stronger germinal center (GC) responses. Specifically, S_Omicron-6P induced even stronger antibody responses against Omicron variants in CoronaVac and Ad5-nCoV-primed animals than non-Omicron-specific vaccines but with limited differences as compared to Ad5-nCoV and S_WT-2P. In addition, boosting with a specific vaccine has the potential to remodel the existing immune profiles. These findings indicated that adenovirus-vectored vaccines and mRNA vaccines would be more effective than other types of vaccines as booster shots in combating Omicron infections. Moreover, the protective efficacies of the vaccines in booster vaccinations are highly related to GC reactions in secondary lymphatic organs. In summary, these findings provide timely important information on prime-boost regimens and future vaccine design.

Objective:To analyze the clinical features and genetic sequencing results of a patient with Kallmann syndrome(KS) carrying digenic mutations who initially presented with osteoporosis, and to enhance awareness of this disease phenotype.Methods:Clinical data were collected, and peripheral blood DNA was extracted for whole-exome sequencing. Relevant literature was reviewed to summarize phenotypes associated with digenic/oligogenic variants involving CHD7.Results:The patient exhibited back pain, delayed development of secondary sexual characteristics, and hyposmia. Laboratory tests revealed reduced sex hormones and gonadotropin levels, while pituitary imaging was unremarkable. Bone mineral density imaging confirmed osteoporosis, and thoracolumbar X-rays showed multiple vertebral compression fractures. Genetic analysis identified a heterozygous splice-site mutation in CHD7(c.2698-1G>T) and a heterozygous missense mutation in WDR11(c.439G>A: p.D147N). According to ACMG criteria, the CHD7 mutation was classified as pathogenic, while the WDR11 variant was defined as a variant of uncertain significance(VUS). Literature review indicated that 40% of KS patients with digenic/oligogenic variants involving CHD7 presented with hearing or ocular abnormalities.Conclusion:This study reports a novel CHD7 mutation and a previously undescribed digenic combination of CHD7 and WDR11 variants in a KS patient. CHD7 variants may be implicated in auditory or ocular involvement in KS cases with digenic/oligogenic inheritances. KS patients may also manifest skeletal abnormalities in addition to hypogonadotropic hypogonadism. Tailored management of sex hormones and osteoporosis therapies across life stages is essential for optimizing bone health in KS.

AIM:This study aims to investigate the role and mechanism of proprotein convertase subtilisin/kexin type 9(PCSK9)in ovarian cancer.METHODS:We compared the expression levels of PCSK9 between ovarian cancer specimens and their corresponding adjacent non-cancerous tissues,while also assessing its expression in various ovarian cancer cell lines.Using a shRNA strategy,we reduced the expression of PCSK9 in ovarian cancer cell lines cul-tured in vitro,with confirmation via Western blot.The effects of PCSK9 downregulation on the proliferation,migration,and invasion of ovarian cancer cells were evaluated through EdU,colony formation,and Transwell assays.Additionally,we analyzed the impact of PCSK9 down-regulation on the MAPK/ERK signaling pathway using Western blot analysis.RE-SULTS:PCSK9 was significantly upregulated in ovarian cancer tissues and cell lines(P＜0.01).Downregulation of PC-SK9 resulted in a significant decrease in cell proliferation,migration,and invasion(P＜0.01).Western blot analysis dem-onstrated that PCSK9 knockdown led to reduced expression levels of key molecules within the MAPK/ERK signaling path-way(P＜0.01).CONCLUSION:PCSK9 promotes the proliferation and invasion of ovarian cancer cells by activating MAPK/ERK signaling pathway.

Objective:To analyze the clinical features and genetic sequencing results of a patient with Kallmann syndrome(KS) carrying digenic mutations who initially presented with osteoporosis, and to enhance awareness of this disease phenotype.Methods:Clinical data were collected, and peripheral blood DNA was extracted for whole-exome sequencing. Relevant literature was reviewed to summarize phenotypes associated with digenic/oligogenic variants involving CHD7.Results:The patient exhibited back pain, delayed development of secondary sexual characteristics, and hyposmia. Laboratory tests revealed reduced sex hormones and gonadotropin levels, while pituitary imaging was unremarkable. Bone mineral density imaging confirmed osteoporosis, and thoracolumbar X-rays showed multiple vertebral compression fractures. Genetic analysis identified a heterozygous splice-site mutation in CHD7(c.2698-1G>T) and a heterozygous missense mutation in WDR11(c.439G>A: p.D147N). According to ACMG criteria, the CHD7 mutation was classified as pathogenic, while the WDR11 variant was defined as a variant of uncertain significance(VUS). Literature review indicated that 40% of KS patients with digenic/oligogenic variants involving CHD7 presented with hearing or ocular abnormalities.Conclusion:This study reports a novel CHD7 mutation and a previously undescribed digenic combination of CHD7 and WDR11 variants in a KS patient. CHD7 variants may be implicated in auditory or ocular involvement in KS cases with digenic/oligogenic inheritances. KS patients may also manifest skeletal abnormalities in addition to hypogonadotropic hypogonadism. Tailored management of sex hormones and osteoporosis therapies across life stages is essential for optimizing bone health in KS.