Objective To prepare a dissolvable microneedle(MN)patch loaded with sodium houttuyfonate(SH)combined with erythromycin(ERY)(SH+ERY-MN patch),to characterize its properties,and to investigate its synergistic anti-acne effects and the mechanisms involved.Methods The SH+ERY-MN patch was prepared by two-step centrifugation.The morphology and skin puncture performance of the microneedles were evaluated using scanning electron microscopy,puncture tests,and other methods.The combined antibacterial effect was assessed using the checkerboard assay.The in vitro anti-inflammatory effect of the SH+ERY-MN patch was evaluated by quantitative PCR(qPCR)and enzyme-linked immunosorbent assay(ELISA).The therapeutic effect of the SH+ERY-MN patch on acnes in mice was investigated using an animal model.Results The microneedle patch prepared in the study had well-aligned microneedles,excellent needle shape,and good mechanical properties,which could effectively penetrate the skin to reach the superficial dermis.The minimum inhibitory concentration(MIC)of ERY against acne was 1.28 mg/mL.When ERY was combined with SH,the results showed that the fractional inhibitory concentration(FIC)was 0.375,indicating that the medication combination had a synergistic antibacterial effect.In particular,the MIC value of ERY in combination was 1/4 of that of ERY used alone.The results of in vitro qPCR showed that SH+ERY-MN could downregulate the production of interleukin(IL)-1β,IL-18,tumor necrosis factor α(TNF-α),inhibitor of κB(I-κB),Toll-like receptor 4(TLR4),NLR family pyrin domain containing 3(NLRP3),and cysteine-aspartic proteases(Caspase-1)induced by Propionibacterium acnes(PA)in cell culture supernatant(P＜0.05).According to the in vivo findings,SH+ERY-MN had a significant anti-acne effect and effectively alleviated cytokine-mediated inflammatory responses.The results of qPCR and ELISA showed that SH+ERY-MN could effectively reduce the levels of IL-1β,IL-18,NLRP3,and Caspase-1 in the serum of the mouse acne inflammation model(P＜0.01),and had good antibacterial and anti-inflammatory effects.Conclusion SH+ERY-MN patch was successfully prepared and demonstrated obvious anti-acne effects.The SH+ERY-MN patch enhances the transdermal delivery of SH combined with ERY.It provides an innovative method for acne treatment and creates new possibilities for achieving effective drug delivery and improving therapeutic efficacy.

Objective:To investigate the expression of potassium calcium-activated channel subfamily N member 4(KCNN4)in pancreatic cancer tissues and its impact on tumor progression,and to explore its role in clinical diagnosis and prognosis evaluation of pancreatic cancer.Methods:Using the GEPIA2 data analysis platform,the expression of KCNN4 in pancreatic cancer tissues and its correlation with patient prognosis were analyzed by integrating data from the TCGA and GTEx databases.Cancerous and adjacent non-cancerous tissues from 24 patients with pancreatic cancer who underwent surgical resection at ChangHai Hospital of the Naval Medical University were collected.The expression of KCNN4 in pancreatic cancer tissues was validated using qPCR,Western blotting,and immunohistochemical staining.The expression of KCNN4 in human pancreatic cancer cell lines BXPC3 and PANC-1 was knocked down using shRNA.CCK-8 and colony formation assays were performed to detect tumor cell proliferation and growth.A murine KPC cell pancreatic cancer model was established to investigate the effects of KCNN4 knockdown on the growth of orthotopic pancreatic tumor and overall survival(OS)in mice.Results:Analysis of TCGA and GTEx data revealed that KCNN4 was highly expressed in pancreatic cancer tissues(P<0.05)and was associated with shortened OS and disease-free survival(DFS)in patients(both P<0.05).The expression levels of KCNN4 mRNA and protein were significantly elevated in pancreatic cancer tissues compared with those in adjacent non-cancerous tissues(all P<0.01).Knockdown of KCNN4 led to significantly reduced growth rates and fewer colony formations in pancreatic cancer cells(both P<0.01).The murine orthotopic pancreatic tumor experiment revealed that KCNN4 knockdown inhibited tumor progression and prolonged the OS of mice.Conclusion:KCNN4,highly expressed in pancreatic cancer tissues,promotes pancreatic cancer cell proliferation and tumor progression,and its expression is closely associated with patient prognosis,suggesting that KCNN4 may serve as a promising target for clinical diagnosis and prognosis evaluation of pancreatic cancer.

Background::Artificial intelligence (AI) technology represented by deep learning has made remarkable achievements in digital pathology, enhancing the accuracy and reliability of diagnosis and prognosis evaluation. The spatial distribution of CD3 + and CD8 + T cells within the tumor microenvironment has been demonstrated to have a significant impact on the prognosis of colorectal cancer (CRC). This study aimed to investigate CD3 CT (CD3 + T cells density in the core of the tumor [CT]) prognostic ability in patients with CRC by using AI technology. Methods::The study involved the enrollment of 492 patients from two distinct medical centers, with 358 patients assigned to the training cohort and an additional 134 patients allocated to the validation cohort. To facilitate tissue segmentation and T-cells quantification in whole-slide images (WSIs), a fully automated workflow based on deep learning was devised. Upon the completion of tissue segmentation and subsequent cell segmentation, a comprehensive analysis was conducted.Results::The evaluation of various positive T cell densities revealed comparable discriminatory ability between CD3 CT and CD3-CD8 (the combination of CD3 + and CD8 + T cells density within the CT and invasive margin) in predicting mortality (C-index in training cohort: 0.65 vs. 0.64; validation cohort: 0.69 vs. 0.69). The CD3 CT was confirmed as an independent prognostic factor, with high CD3 CT density associated with increased overall survival (OS) in the training cohort (hazard ratio [HR] = 0.22, 95% confidence interval [CI]: 0.12–0.38, P <0.001) and validation cohort (HR = 0.21, 95% CI: 0.05–0.92, P = 0.037). Conclusions::We quantify the spatial distribution of CD3 + and CD8 + T cells within tissue regions in WSIs using AI technology. The CD3 CT confirmed as a stage-independent predictor for OS in CRC patients. Moreover, CD3 CT shows promise in simplifying the CD3-CD8 system and facilitating its practical application in clinical settings.