ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes（TP-CX@TESs）， conduct its quality evaluation， and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators， Box-Behnken design-response surface methodology（BBD-RSM） was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide（LPS）-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention， the levels of inflammatory factors such as nitric oxide（NO）， interleukin-6（IL-6）， and tumor necrosis factor-α（TNF-α） in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2（JAK2）， signal transducer and activator of transcription 3（STAT3）， and α7 nicotinic acetylcholine receptor（α7nAChR）， and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was applied to measure the mRNA expression levels of JAK2， STAT3， the encoding gene of α7nAChR（CHRNA7）， and nuclear transcription factor-κB（NF-κB）. ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows：egg yolk lecithin content of 2.3%， ethanol volume fraction of 30%， and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of （105.60±3.85） nm， a polydispersity index of 0.19±0.03， and a Zeta potential of （-15.89±0.98） mV. The encapsulation efficiencies of triptolide， ferulic acid， and ligustilide were （76.88±4.40）%， （78.84±4.40）%， and （65.88±0.06）%， respectively. Both in vitro release and transdermal penetration of triptolide， ferulic acid， and ligustilide in TP-CX@TESs all followed the first-order kinetic model， showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO， TNF-α， and IL-6（P<0.01）， remarkably upregulated the protein expression levels of STAT3 and α7nAChR（P<0.01）， increased the mRNA expression level of CHRNA7， and significantly downregulated the mRNA expression level of NF-κB（P<0.05， P<0.01）. ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible， along with favorable in vitro release property， good transdermal permeability， and excellent in vitro anti-inflammatory activity， the mechanism is related to the inhibition of NF-κB.

ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes（TP-CX@TESs）， conduct its quality evaluation， and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators， Box-Behnken design-response surface methodology（BBD-RSM） was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide（LPS）-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention， the levels of inflammatory factors such as nitric oxide（NO）， interleukin-6（IL-6）， and tumor necrosis factor-α（TNF-α） in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2（JAK2）， signal transducer and activator of transcription 3（STAT3）， and α7 nicotinic acetylcholine receptor（α7nAChR）， and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was applied to measure the mRNA expression levels of JAK2， STAT3， the encoding gene of α7nAChR（CHRNA7）， and nuclear transcription factor-κB（NF-κB）. ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows：egg yolk lecithin content of 2.3%， ethanol volume fraction of 30%， and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of （105.60±3.85） nm， a polydispersity index of 0.19±0.03， and a Zeta potential of （-15.89±0.98） mV. The encapsulation efficiencies of triptolide， ferulic acid， and ligustilide were （76.88±4.40）%， （78.84±4.40）%， and （65.88±0.06）%， respectively. Both in vitro release and transdermal penetration of triptolide， ferulic acid， and ligustilide in TP-CX@TESs all followed the first-order kinetic model， showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO， TNF-α， and IL-6（P<0.01）， remarkably upregulated the protein expression levels of STAT3 and α7nAChR（P<0.01）， increased the mRNA expression level of CHRNA7， and significantly downregulated the mRNA expression level of NF-κB（P<0.05， P<0.01）. ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible， along with favorable in vitro release property， good transdermal permeability， and excellent in vitro anti-inflammatory activity， the mechanism is related to the inhibition of NF-κB.

ObjectiveTo verify the clinical efficacy of Qingfei Huatan Tongqiao Formula in treating children with adenoid hypertrophy （AH） of the lung-heat obstructing the nose syndrome， and to observe its effects on quality of life. MethodsA multicenter， double-blind， double-dummy， randomized controlled clinical trial was conducted. A total of 160 pediatric AH outpatients meeting the inclusion and exclusion criteria were enrolled between November 2022 and November 2024 from Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine， Shanghai Municipal Hospital of Traditional Chinese Medicine， Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine， and Yueyang Hospital of Integrated Traditional Chinese and Western Medicine. The children were randomly assigned to a treatment group （80 cases） or a control group 80 cases）. The treatment group received Qingfei Huatan Tongqiao Formula combined with a montelukast sodium placebo， while the control group received a Qingfei Huatan Tongqiao Formula placebo combined with montelukast sodium. The treatment course lasted 90 days. After treatment， therapeutic efficacy， sleep monitoring parameters， and Obstructive Sleep Apnea-18 （OSA-18） scores were compared between groups. ResultsCompared with pre-treatment values within each group， both groups showed significant decreases in the adenoid-to-nasopharyngeal ratio （A/N）， primary symptom scores， and TCM syndrome scores. OSA-18 scores were significantly reduced. Sleep monitoring showed decreases in the Apnea-Hypopnea Index （AHI） and Oxygen Desaturation Index （ODI）， and an increase in minimum SpO₂ （all P<0.05）. After treatment， compared with the control group， the treatment group had a lower A/N ratio and higher overall effectiveness rate and TCM syndrome effectiveness rate. The treatment group also had lower total OSA-18 scores and lower subscale scores （sleep disturbance， physical symptoms， emotional distress， and caregiver impact）， with statistically significant differences （all P<0.05）. There was no significant difference in daytime symptoms （P=0.201）. Regarding sleep monitoring indices， the treatment group had lower AHI and ODI and higher minimum SpO₂ （all P<0.05）， with no significant difference in average SpO₂ （P=0.116）. ConclusionQingfei Huatan Tongqiao formula can significantly reduce adenoid size in children with AH， alleviate clinical symptoms， improve quality of life， reduce sleep-disordered breathing events， and ameliorate hypoxia. Its therapeutic efficacy is significantly superior to that of montelukast sodium.

ObjectiveTo investigate the effects of the combination of components from Tripterygii Radix et Rhizoma and Chuanxiong Rhizoma on rheumatoid arthritis fibroblast-like synoviocytes （RA-FLSs） and the underlying mechanism. MethodsRA-FLSs were grouped as follows： blank control， positive control （methotrexate）， Tripterygii Radix et Rhizoma components， Chuanxiong Rhizoma components， and components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma. The cell-counting kit-8 （CCK-8） assay was employed to the cell proliferation， invasion， and apoptosis. The levels of tumor necrosis factor （TNF）-α， interleukin （IL）-6， reactive oxygen species （ROS）， and malondiadehyde （MDA） in cells were measured. Western blot was employed to determine the protein levels of nuclear factor erythroid 2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， nuclear factor-kappa B （NF-κB） p65， phosphorylated inhibitory subunit of NF-κBα （p-IκBα）， cysteinyl aspartate-specific protease-3 （Caspase-3）， and B-cell lymphoma 2 （Bcl-2）. Real-time PCR was employed to determine the mRNA levels of Nrf2， HO-1， and NF-κB p65. ResultsThe cells in the groups of positive control， Tripterygii Radix et Rhizoma components， Chuanxiong Rhizoma components， and components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma were treated with 2.50 mg·L^-1 methotrexate， 0.20 mg·L^-1 triptolide + 0.20 mg·L^-1 celastrol， 5.00 mg·L^-1 ferulic acid + 20.00 mg·L^-1 ligustrazine， 0.20 mg·L^-1 triptolide + 0.20 mg·L^-1 celastrol + 5.00 mg·L^-1 ferulic acid + 20.00 mg·L^-1 ligustrazine， respectively. Compared with the blank control group， drug administration reduced the proliferation and invasion and increased the apoptosis of cells （P<0.01）， lowered the levels of TNF-α， IL-6， ROS， and MDA （P<0.01）， up-regulated the mRNA and protein levels of Caspase-3， Nrf2， and HO-1 （P<0.01）， and down-regulated the mRNA and protein levels of Bcl-2， NF-κB p65， and p-IκBα （P<0.01）. Compared with the Tripterygii Radix et Rhizoma components group， the combination of components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma inhibited the proliferation and invasion （P<0.05） and promoted the apoptosis of RA-FLSs， up-regulated the mRNA levels of Nrf2 and HO-1 and protein levels of Nrf2 and Caspase-3 （P<0.05）， and down-regulated the protein levels of NF-κB p65 and p-IκBα （P<0.05）. ConclusionThe combination of components from Chuanxiong Rhizoma and Tripterygii Radix et Rhizoma can inhibit the proliferation and invasion and promote the apoptosis of RA-FLSs and alleviate oxidative stress and inflammation by inhibiting the NF-κB signaling pathway， activating the Nrf2/HO-1 pathway， and regulating the expression of Bcl-2/Caspase-3.

Objective : To explore the effect of Wilms′ tumor 1-associated protein(WTAP) on tissue collagen deposition in pulmonary fibrosis caused by bleomycin. Methods : 60 mice were randomly divided into four groups: control group(Control group), Bleomycin-induced pulmonary fibrosis group(BLM group), pulmonary fibrosis lentivirus empty vector control group(BLM+LV-NC group), pulmonary fibrosis WTAP lentivirus group virus group(BLM+LV-WTAP group). Experimental pulmonary fibrosis mouse model was established by subcutaneous injection of bleomycin(35 mg/kg) into the abdomen, twice a week for a total of 8 times. After modeling, Western Blot was used to detect the protein expression of fibrosis-related markers α-smooth muscle actin(α-SMA), type I collagen(Collagen Ⅰ), fibronectin(Fibronectin), and WTAP protein. Masson staining and Sirius Red staining were used to detect collagen deposition. RT-qPCR was used to detect WTAP mRNA expression, WTAP lentivirus infection effect, and Collagen Ⅰ mRNA expression. Results: Compared with the Control group, the expression of pulmonary fibrosis markers α-SMA(P<0.001), Collagen Ⅰ(P<0.001), and Fibronectin(P<0.01) protein in the BLM group all increased. Masson staining(P<0.001) and Sirius Red staining(P<0.001) confirmed that significant collagen deposition occurred in the lung tissue of the BLM group. In addition, the expression of WTAP protein in the lung tissue of the BLM group increased(P<0.01). Compared with the Control group, the expression of WTAP mRNA in the BLM group increased(P<0.001). Compared with the BLM+LV-NC group, the expression of WTAP mRNA in the tissues of the BLM+LV-WTAP group decreased(P<0.001), proving that virus infection is effective. After infection with WTAP lentivirus, collagen fiber deposition decreased(P<0.001), Collagen Ⅰ mRNA(P<0.001) level decreased, and protein(P<0.001) expression decreased in the BLM+LV-WTAP group. Conclusion Knocking down of WTAP can reduce collagen deposition in bleomycin-induced pulmonary fibrosis tissue in mice and improve experimental pulmonary fibrosis.

Objective: To investigate the effect of sirtuin 2(SIRT2) on the proliferation and migration of cardiac fibroblasts(CFs)in C57BL/6 mice under angiotensin II(Ang Ⅱ) stimulation. Methods : The hearts were taken from 1 to 2 days C57BL/6 milk mice. After cutting and digesting, CFs were extracted by different adhesion centrifugation. After CFs attachment, the cells were cultured under control medium and Ang Ⅱ(100 nmol/L) medium and treated using OE-SIRT2 plasmid to overexpression the SIRT2 gene. RT-qPCR was used to detect mRNA expression of SIRT2 proliferating cell nuclear antigen(PCNA), periostin(POSTN)and type Ⅰ collagen procollagen A1(Col1A1), Western blot assay was used to measure the protein expression levels of SIRT2, PCNA, POSTN and Col1A1, CCK-8 assay and EdU assay were used to evaluate CFs proliferation rate, Transwell experiment was used to assess CFs migration activity. Results: Compared with control group, Ang Ⅱ stimulation led to down-regulation of SIRT2 expression in CFs, increased collagen expression, and promoted CFs proliferation and migration. The expression of SIRT2 was up regulated in CFs treated with OE-SIRT2 plasmid under Ang Ⅱ stimulation, Col1A1, POSTN and PCNA expression was down regulated, and CFs proliferation and migration ability decreased. Conclusion Overexpression of SIRT2 can inhibit the proliferation and migration of CFs under Ang Ⅱ stimulation, indicating that SIRT2 may be a key regulatory point in the onset and progression of cardiac fibrosis.

With the further comprehensive understanding of pathologists in the morphological characteristics of DICER1-related thyroid tumors,and the widespread application of multi-gene detection technology,the detection rate of these tumors is gradually increasing.DICER1 mutations can be detected in benign,low-risk,and malignant thyroid tumors.It is particularly noteworthy that tumors with germline mutations and in pediatric patients often exhibit more ag-gressive and prone to recurrent.Therefore,it is important to identify and diagnose these lesions.This article summarize the advancements in DICER1 mutation-related thyroid tumors through systematic interpretation.

Significant advancements have been achieved in both basic and clinical research pertaining to thyroid cancer,leading to substantial updates in clinical diagnostic and therapeutic protocols and guidelines for various thyroid cancer subtypes.The advent and widespread application of next-generation sequencing technology in clinical practice have brought protein kinase fusion-driven thyroid cancer into sharper focus.This particular subtype accounts for over 10％of papillary thyroid carcinomas and frequently exhibits more aggressive histological characteristics and biological behaviors.The identification of kinase fusion-related thyroid cancer and the implementation of optimal detection strate-gies pose notable challenges for pathologists in China.This article provides a comprehensive review of the molecular mechanisms underlying phosphorylation signal transduction driven by kinase fusion,the pathological characteristics and detection methodologies of kinase fusion-related thyroid cancer,as well as the progress in research on related targeted therapies.The aim is to enhance the understanding of kinase fusion-related thyroid cancer,with the aspiration of facili-tating personalized and optimal diagnostic and therapeutic strategies in daily clinical practice,ultimately maximizing pa-tient outcomes.

Purpose To summarize the clinicopathological features of pheochromocytoma and paraganglioma(PPGL)and discuss the potential correlation between SDHB immunophenotype and prognosis in PPGLs.Methods A retrospective analysis was conducted on 30 samples of PPGL along with their corresponding clinicopathological informa-tion,SDHB immunophenotype characteristics,and the risk of recurrence and metastasis.Results The study included 20 extra-adrenal paragangliomas and 10 pheochromocytoma cases.The male-to-female ratio was 13∶17,with a mean age of 56(range from 21 to 79).Four cases recurred,one case resulted in death and five cases failed to follow-up.All recurrent or fatal cases were paraganglioma patients.Among the 30 cases,3 had multiple nodular lesions,and the re-maining cases were single nodule.The neck was the most frequent site for paraganglioma(6/20),followed by retroper-itoneum(5/20).Histologically,the tumors displayed a variable"zellballen"architecture with a highly vascularized stroma.The chief cells had abundant pale eosinophilic cytoplasm and slightly to moderately atypical nuclei,and pe-ripherally located sustentacular cells.Positive immunoreactivity with markers of neuroendocrine cells,including Syn,CgA,and GATA3,was found in tumor chief cells,which were nonreactive for CK.The sustentacular cells exhibited positive immunoreactivity for the S-100 protein.SDHB deficiency was demonstrated in 12 of 30 cases,with only one case being pheochromocytoma.The recurrence rate in SDHB-deficient group was higher than that in the positive group(33.3％vs 6.7％).Only one case of paraganglioma developed distant metastasis and death.Conclusion SDHB de-ficiency was predominantly observed in paragangliomas and serverd as an indipentent factor for metastatic risk in PPGLs.It was closely associated with younger age at onset,invasiveness,extra-adrenal tumorgenesis,and a high rate of tumor recurrence.

Objective To explore the effect of lipoprotein a[Lp(a)]on the inflammatory state and cardiovascular system of patients with essential hypertension.Methods The clinical data of 234 patients with essential hypertension admitted from January 2022 to November 2023 were retrospectively collected.According to the serum Lp(a)concentration,they were divided into the normal Lp(a)group(n=185)and the high Lp(a)group(n=49).The differences in serum inflammatory markers C reactive protein(CRP),neutrophil-to-lymphocyte ratio(NLR),and fibrinogen between the two groups were compared.Pearson correlation analysis was used to analyze the correlation between Lp(a)and inflammatory markers,and the incidence of cardiac target organ damage between the two groups of patients was compared.Results Compared with the normal Lp(a)group,the CRP concentration,NLR and fibrinogen concentration of hypertensive patients in the high Lp(a)group increased(P<0.05,P<0.01),and the concentration of Lp(a)was positively correlated with CRP,NLR and fibrinogen(r=0.168,0.165,0.321,P<0.05).The incidence of myocardial infarction was higher in the high Lp(a)group[22.45%(11/49)]than in the normal Lp(a)group[7.57%(14/185)](P<0.01).Conclusion In patients with essential hypertension,high Lp(a)concentration is associated with a stronger inflammatory response,and elevated Lp(a)concentration is related to an increased incidence of myocardial infarction,suggesting that Lp(a)may affect target organ damage in patients with essential hypertension by promoting inflammation.