Identification of Fangchinoline as a novel autophagy inhibitor with an adjuvant of chemotherapy against lung cancer.

Autophagy is a fundamental recycling pathway that enhances cellular resilience, promoting survival. However, this survival mechanism can impede anti-cancer treatment strategies designed to induce cell death. In this study, we identified a novel autophagy inhibitor, Fangchinoline (Fan) isolated from the traditional Chinese medicine Stephania tetrandra. We speculated that when Fan blocks autophagy, cancer cells lose substantial self-preservation abilities during treatment. Firstly, we examined in detail the mechanism through which Fan inhibits autophagy. Specifically, Fan induced a significant increase in autophagosomes, as indicated by GFP-LC3 labeling, confirmed by the up-regulation of LC3-II. The autophagy receptor protein p62 was also up-regulated, suggesting a potential inhibition of autophagy flux. We further ruled out the possibility of fusion barriers between lysosomes and autophagosomes, as confirmed by their co-localization in double fluorescence staining. However, the lysosomal acid environment might be compromised, as suggested by the diminished fluorescence of acidity-sensitive dyes in the lysosomes and the corresponding decrease in mature forms of lysosomal cathepsin. To test the anti-cancer potential of Fan, we combined it with Cisplatin (Cis) or Paclitaxel (PTX) for lung cancer cell treatment. This combined treatment demonstrated a synergistically enhanced killing effect. These promising anti-tumor results were also replicated in a xenografted tumor model. The significance of this research lies in the identification of Fan as a potent autophagy inhibitor and its potential to enhance the efficacy of existing anti-cancer drugs. By unraveling the mechanisms of Fan's action on autophagy and demonstrating its synergistic effect in combination therapies, our study provides valuable insights for developing novel strategies to overcome autophagy-mediated resistance in cancer treatment.

Identification of Fangjihuangqi Decoction as a late-stage autophagy inhibitor with an adjuvant anti-tumor effect against non-small cell lung cancer.

BACKGROUND: Clinically, although chemotherapy is one of the most commonly used methods of treating tumors, chemotherapeutic drugs can induce autophagic flux and increase tumor cell resistance, leading to drug tolerance. Therefore, theoretically, inhibiting autophagy may improve the efficacy of chemotherapy. The discovery of autophagy regulators and their potential application as adjuvant anti-cancer drugs is of substantial importance. In this study, we clarified that Fangjihuangqi Decoction (FJHQ, traditional Chinese medicine) is an autophagy inhibitor, which can synergistically enhance the effect of cisplatin and paclitaxel on non-small cell lung cancer (NSCLC) cells. METHODS: We observed the changes of autophagy level in NSCLC cells under the effect of FJHQ, and verified the level of the autophagy marker protein and cathepsin. Apoptosis was detected after the combination of FJHQ with cisplatin or paclitaxel, and NAC (ROS scavenger) was further used to verify the activation of ROS-MAPK pathway by FJHQ. RESULTS: We observed that FJHQ induced autophagosomes in NSCLC cells and increased the levels of P62 and LC3-II protein expression in a concentration- and time-gradient-dependent manner, indicating that autophagic flux was inhibited. Co-localization experiments further showed that while FJHQ did not inhibit autophagosome and lysosome fusion, it affected the maturation of cathepsin and thus inhibited the autophagic pathway. Finally, we found that the combination of FJHQ with cisplatin or paclitaxel increased the apoptosis rate of NSCLC cells, due to increased ROS accumulation and further activation of the ROS-MAPK pathway. This synergistic effect could be reversed by NAC. CONCLUSION: Collectively, these results demonstrate that FJHQ is a novel late-stage autophagy inhibitor that can amplify the anti-tumor effect of cisplatin and paclitaxel against NSCLC cells.

Effect of Static Magnetic Field Assisted Thawing on Physicochemical Quality and Microstructure of Frozen Beef Tenderloin.

Although freezing is the most common and widespread way to preserve food for a long time, the accumulation of microstructural damage caused by ice crystal formation during freezing and recrystallization phenomena during thawing tends to degrade the quality of the product. Thus, the side effects of the above processes should be avoided as much as possible. To evaluate the effect of different magnetic field strength assisted thawing (MAT) on beef quality, the indicators associated with quality of MAT-treated (10-50 Gs) samples and samples thawed without an external magnetic field were compared. Results indicated that the thawing time was reduced by 21.5-40% after applying MAT. Meat quality results demonstrated that at appropriate magnetic field strengths thawing loss, TBARS values, cooking loss, and shear force were significantly decreased. Moreover, by protecting the microstructure of the muscle, MAT significantly increased the a∗ value and protein content. MAT treatment significantly improved the thawing efficiency and quality of frozen beef, indicating its promising application in frozen meat thawing.

[Effect of tympanoplasty on wet ear with different mucosal status of tympanic cavity].

Objective:A retrospective analysis of audiologic outcome and graft take rate on post-tympanoplasty with different middle ear mucosal conditions in wet ear. Method:According to the characteristics of middle ear mucosal condition and residual eardrum, 80 cases with wet ear of chronic suppurative otitis media were divided into the hydrocele group, the swelling group and the granulation group. The factors in different groups, including gender, age, disease course, sides, size and location of perforations, destruction of ossicular chain and reconstruction methods were analyzed. Moreover, postoperative hearing improvement and graft take rate were compared among the three groups. Result:There was no significant difference in gender, age, disease course, sides, size and location of perforations among the hydrocele group, the swelling group and the granulation group （P>0.05）. Overall, the postoperative average Air-Bone Gaps（ABG） were reduced in all wet ear patients after surgery （P<0.01）. The ABG was decreased from （25.5 ± 10.8） dB to（15.4 ± 9.4） dB in the hydrocele group, and decreased from （27.6 ± 8.7） dB to （15.2 ± 9.6） dB in the swelling group, and from （29.5 ± 7.7） dB to （17.2 ± 17.2） dB in the granulation group. The graft take rates were 90.0% in totally. There were no significant difference in graft take rates among the three groups, and 84.6% in the hydrocele group, 93.3% in the swelling group and 100.0% in the swelling group（P>0.05）. Conclusion:Wet ear is not an absolute contraindication of tympanoplasty for chronic suppurative otitis media. Whether there was effusion, swelling or granulomatous hyperplasia in the tympanoplasty, the patients'hearing improved significantly after tympanoplasty, and the healing rate of the tympanoplasty did not decrease. Further basic and clinical studies are needed to standardize the timing of wet ear surgery, clarify the operative contraindication and elucidate the pathophysiological mechanism of eardrum healing.