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.

Medullary thyroid carcinoma (MTC) is the most common neuroendocrine carcinoma within the thyroid gland, characterized by strong invasiveness, high metastasis and recurrence rates. It is relatively rare among thyroid malignancies. The cytological and histological features of MTC are diverse and disperse, presenting as papillary, follicular, solid, trabecular, and spindle cell patterns. Immunohistochemical staining shows variable expression of calcitonin, carcinoembryonic antigen, and neuroendocrine markers. MTC can be classified into hereditary and sporadic types, with most cases caused by germline or somatic mutations in the RET gene located on chromosome 10. The 5th edition World Health Organization classification of endocrine and neuroendocrine tumors categorizes MTC into low-grade and high-grade based on tumor necrosis, mitotic figures, and Ki-67 proliferation index, highlighting that histological grading and RET gene mutations are independent prognostic predictors. This paper summarizes the recent advances in the pathological diagnosis of MTC, focusing on the key roles of the MTC grading system, molecular characteristics, and genetic screening and counseling in risk stratification for recurrence and targeted therapy.

Objective:To investigate the cytopathological features of thyroid tumor with DICER1 mutation.Methods:A retrospective study on the preoperative cell smear was conducted on thyroid tumors with DICER1 gene mutations detected by Sanger sequencing in the Department of Pathology Shanghai Sixth People′s Hospital affiliated to Shanghai Jiaotong University School of Medicine from May 2022 to November 2024.Results:Totally 163 cases with histological features indicating DICER1 mutation related thyroid tumor underwent Sanger sequencing. Fifteen cases were confirmed to harbor DICER1 mutation (15/163,9.2%). Fourteen of 15 patients were female, and only 1 was male; average age 42(31,47) years. Eight cases presented with D1709 hotspot mutation (8/15) and 7 cases with the E1813 hotspot mutation (7/15) and there was no statistical significant difference between mutation rate of different hotspot ( F=0.620， P=0.438). All specimens were stained with hematoxylin-eosin staining. A moderate number of cells were observed for all cases, predominantly with macrofollicular pattern and rare small papillae. The cell nuclei were mainly uniform, small, round and dark, slightly enlarged or medium-sized. Several cases could also present RAS-like nuclear features: 3 cases showed visible nuclear grooves. According to the expert consensus on the cytopathological diagnosis of thyroid fine needle aspiration (version 2023),the cytopathological diagnostic categories were: Ⅱ, 6 cases; Ⅲ, 2 cases; Ⅳ, 6 cases; and Ⅴ, 1 case. Postoperative histological diagnoses included follicular thyroid carcinoma in 2 cases, high-grade differentiated thyroid carcinoma in 1 case, follicular thyroid adenoma in 3 cases, follicular thyroid tumor of undetermined malignant potential in 4 case, and thyroid follicular nodular disease in 5 cases. Conclusions:The morphological features of DICER1-mutated thyroid tumors are predominant macrofollicles, with uniformly small round and dark nuclei. It is difficult to identify typically diagnostic atrophic follicles on cell smear, leading to a preoperative diagnosis of benign non-neoplastic or indeterminate category. Therefore, it is necessary to carefully observe the macro-follicles and small round dark nuclear features, which is necessary to suggest a genetic test of DICER1 gene and to confirm the diagnosis before surgery.

Objective:To investigate the clinical, radiological and histological features of intraosseous hibernoma.Methods:Two cases of intraosseous hibernoma diagnosed in the Department of Pathology, the Sixth People′s Hospital Affiliated to Shanghai Jiaotong University School of Medicine from 2020 to 2023 were analyzed. Related literature was also reviewed.Results:One case was a 44-year-old female with abnormal signals in the proximal right femur revealed by MRI, who underwent curettage of the lesion. The other case was a 41-year-old female with an occupying lesion in the second sacral vertebrae revealed by CT and MRI, who underwent CT-guided biopsy. Microscopically, both tumors were composed of large polygonal cells, with finely vacuolated cytoplasm and distinct cell membranes and variably admixed mature adipose cells or hematopoietic components within the stroma. Nuclei were small, centrally or paracentrally situated and displayed prominent scalloping. Nuclear atypia was absent. A relatively clear tumor boundary could be observed in one case. Immunohistochemical staining showed that tumor cells were positive for S-100 protein, while negative for keratin, CD68, H3F3A, and Brachyury.Conclusions:Intraosseous hibernoma is extremely rare and tends to affect middle-aged and elderly patients. It most frequently occurs in the spine and pelvis. It needs to be differentiated from metastatic cancer, Erdheim-Chester disease, intraosseous lipoma with necrosis, and benign notochordal cell tumor. Increasing awareness of it helps to avoid missed diagnoses or excessive treatment due to misdiagnosis.

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.

Objective To investigate the effects of Aurora kinase A(AURKA)on the tumor microenvironment of colorectal cancer(CRC)and to predict the active compounds in Chinese herbs that can target AURKA.Methods Based on the transcriptomic data and clinical information from 380 CRC tissues and 51 paracancerous tissues in The Cancer Genome Atlas(TCGA)database,the infiltration of different cells in the tumor tissues was analyzed using xCell and the binding of active compounds of Chinese herbs with AURKA was predicted through molecular docking.Results The expression of AURKA was significantly upregulated in CRC tissues compared with that in paracancerous tissues(P＜0.05),and CRC patients with high AURKA expression had shorter overall survival.Compared with the AURKA low-expression group,the abundance of macrophages,monocytes,and effector memory CD4+and CD8+T cells was significantly downregulated in the AURKA high-expression group(P＜0.05).In addition,the cytotoxicity of T cells was significantly reduced(P＜0.05).Further analysis revealed that AURKA expression was positively correlated with the abundance of myeloid-derived suppressor cells(MDSCs)and the expression levels of their chemokines CXCL2 and CXCL5(P＜0.05).Genes that were differentially expressed between the AURKA high-and low-expression groups were mainly enriched in monocyte migration,chemokine-induced cellular responses,and other related processes.Chinese herbal compounds,including hesperidin,aristololactam A Ⅱ a,anacardic acid,coumestrol,and 17β-estradiol,all showed binding energies to AURKA lower than-1.2 kcal/mol,indicating a certain level of binding stability.Among these Chinese herbal compounds,17β-estradiol exhibited the best binding stability to AURKA-3UOL.Conclusion The high expression of AURKA in CRC tissues suggests a poor clinical prognosis.AURKA can promote the development of a suppressive immune microenvironment in CRC,and 17β-estradiol,an active compound from Chinese herbs,is a potential therapeutic agent targeting AURKA.

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.