Effect of Yiqi Fuzheng Jiedu Decoction on Autophagy and Growth of A549 Cells： An Exploration Based on p53/AMPK Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of Yiqi Fuzheng Jiedu decoction （YQFZJDD） on autophagy and growth of A549 cells. Method:A549 cells were intervened with YQFZJDD-containing serum prepared in advance. The levels of microtubule-associated protein 1 light chain 3 （LC3）， homologue of yeast autophagy-related gene 6 （Beclin1）， p62， p53， adenosine 5'-monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， mammalian target of rapamycin （mTOR）， and phosphorylated mTOR （p-mTOR） were detected by Western blot assay， and microtubule-associated protein 1A/1B light chain 3B （MAP1LC3B） by immunofluorescence （IF） assay. The proliferation， invasion， and senescence of A549 cells were separately measured by 5-ethynyl-2'-deoxyuridine （EDU） staining， Transwell assay， and <italic>β</italic>-galactosidase staining. In the presence of autophagy inhibitor 3-methyladenine （3-MA， 5 mmol·L^-1）， the cells were divided into the 10% fetal bovine serum （blank） group， 10% control serum （control） group， low-， medium-， and high-dose 10% YQFZJDD-containing serum groups， and high-dose 10% YQFZJDD-containing serum + 3-MA group， followed by the measurement of A549 cell proliferation， invasion， and senescence. In the adoption of p53 inhibitor Pifithrin-<italic>α</italic> （PFT-<italic>α</italic>， 10 μmol·L^-1）， the cells were divided into the control group， PFT-<italic>α</italic> group， high-dose YQFZJDD-containing serum group， and high-dose YQFZJDD-containing serum + PFT-<italic>α</italic> group. Then the LC3-Ⅱ and Beclin1 expression， MAP1LC3B fluorescence intensity， as well as A549 cell proliferation， invasion and senescence were determined. Result:Compared with the blank group and control group， YQFZJDD-containing serum at the medium and high doses up-regulated the protein expression levels of LC3-Ⅱ and Beclin1 in A549 cells after 48-h intervention in a dose-dependent manner （<italic>P</italic><0.01）. Besides， YQFZJDD-containing serum at the low-， medium-， and high-doses down-regulated p62 and p-mTOR/mTOR expression （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated p53 and p-AMPK/AMPK （<italic>P</italic><0.01）， decreased the number of proliferative and invasive cells， increased the number of senescent cells （<italic>P</italic><0.01）， and enhanced the IF intensity of MAP1LC3B， with the optimal effect observed in the high-dose YQFZJDD-containing serum group （<italic>P</italic><0.01）. Compared with the high-dose YQFZJDD-containing serum group， the high-dose YQFZJDD-containing serum + 3-MA group and high-dose YQFZJDD-containing serum + PFT-<italic>α</italic> group exhibited significantly increased proliferative and invasive cells but decreased senescent cells （<italic>P</italic><0.05， <italic>P<</italic>0.01）. Meanwhile， the IF intensity of MAP1LC3B in the high-dose YQFZJDD-containing serum + PFT-<italic>α</italic> group was weakened and the LC3-Ⅱ and Beclin1 protein expression levels declined （<italic>P</italic><0.05， <italic>P<</italic>0.01）. Conclusion:YQFZJDD promotes the autophagy of A549 cells through the p53/AMPK signaling pathway to inhibit their proliferation， invasion and migration but accelerate senescence， thus playing a crucial role in inhibiting the progression of lung cancer.

Advance in Cricopharyngeal Dysfunction (review) / 中国康复理论与实践

Cricopharyngeus muscle (CPM) is an important component of upper esophageal sphincter. It is termed cricopharyngeus dysfunction (CPD) if CPM fails to relax, which may be second to neurologic or neoplastic disease. However, the pathogenesis is not yet clear. The diagnosis of CPD would combine clinical features and swallowing evalutions, such as videofluoroscopic swallowing study, pharyngeal manometry and electromyogram, etc. Esophageal manometry is not the golden standard and should be combined with videofluoroscopic swallowing study. Interventions include swallowing exercise, pharyngoesophageal balloon dilatation, Botulinum toxin injection and cricopharyngeus myotomy. The outcomes will be better in those with intact laryngeal elevation and pharyngeal contraction. Balloon dilation is more commonly used. The success rate of cricopharyngeus myotomy is reported higher than that of Botulinum toxin injection. Each of the three technologies has their advantages, and it needs more researches to study their indications, contraindications and patient selection.