ABSTRACT
OBJECTIVE To investigate the effect and mechanism of gracillin from Reineckia carnea on autophagy in non- small cell lung cancer A549 cells. METHODS Using A549 cells as subjects， the effects of different concentrations of gracillin （0.25， 0.5， 1， 2， 4 μmol/L） on the proliferation of cells were detected by CCK-8 after being treated for different time （12， 24， 48 h）. Compared with the control group without medication， the effect of gracillin （2 μmol/L） on the formation of autophagosomes in cells was observed by transmission electron microscope after 24 h of exposure. The aggregation of GFP-LC3 on autophagosome membrane was detected by GFP-LC3 plasmid transfection after being treated with gracillin （0.25， 0.5， 1， 2 μmol/L） for 24 h. Quantitative real-time PCR and Western blot assay were used to detect the mRNA and protein expressions of family with sequence similarity 102 member A（FAM102A）， the expressions of autophagy-related proteins [p62， Beclin-1， microtubule-associated protein 1 light chain 3B （LC3B）]， and the expressions of phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway-related proteins in A549 cells after being treated with gracillin （0.25， 0.5， 1 and 2 μmol/L） for 24 h. RESULTS Gracillin significantly inhibited the proliferation of A549 cells in a concentration- and time-dependent manner. The IC50 was 2.55 μmol/L at 24 h. After 24 h of gracillin treatment， autophagosomes with bilayer membrane structure were found in the cell cytoplasm， and GFP-LC3 green fluorescent spots on autophagosome membrane were obvious， representing an increasing trend as drug concentration. Compared with the control group， mRNA and protein expressions of FAM102A （0.5， 1， 2 μmol/L groups）， protein expression of Beclin-1 （1， 2 μmol/L groups） and LC3B-Ⅱ/LC3B-Ⅰ ratio （2 μmol/L group） were significantly increased in different concentrations of gracillin groups， while the protein expression of p62 （1， 2 μmol/L groups）， and the protein phosphorylations of Akt （1， 2 μmol/L groups） and PI3K （2 μmol/L group） were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Gracillin can promote excessive autophagy in A549 cells by up-regulating mRNA and protein expressions of FAM102A and inhibiting PI3K/Akt signaling pathway， thus inhibiting cell proliferation.
ABSTRACT
ObjectiveIn the treatment of acute gouty arthritis （AGA）， western medicine is mostly used for anti-inflammatory and analgesic purposes to control the blood uric acid level， but some patients are still at risk of poor control and recurrent attacks. Chinese medicinal prescriptions， potent in resisting inflammation and relieving pain， are able to stabilize the blood uric acid level， reduce acute attacks， and improve the clinical efficacy of western medicine. However， there is a lack of evidence to support their use as evidence-based medicine. This study employed network Meta-analysis （NMA） to evaluate the efficacy and safety of common Chinese medicinal prescriptions in the treatment of AGA， aiming to provide evidence-based medical evidence for the clinical use of Chinese medicinal prescriptions in the treatment of AGA. MethodChinese and English databases were searched for prospective cohort studies and randomized controlled trials （RCTs） on Chinese medicinal prescriptions against AGA from database inception to December 1， 2022. Stata software and Review Manager were used for statistical analysis. ResultForty-four papers with 3 564 cases involved were included in the current NMA. In terms of reducing blood uric acid， the cumulative probability results showed that Mahuang Lianyao Chixiaodou Tang showed optimal efficacy （87.60%）. In terms of relieving joint pain， Danggui Niantongtang and Guizhi Shaoyao Zhimutang showed optimal efficacy （92.00% and 82.30%）. In terms of improving erythrocyte sedimentation rate （ESR）， Simiaowan was superior to other prescriptions （87.00%）. In terms of reducing C-reactive protein （CRP）， Simiaowan and Baihutang modified with Guizhitang showed superior efficacy （76.00% and 66.10%）. In terms of safety， except for the basic treatment group， Mahuang Lianyao Chixiaodou Tang had the lowest probability of adverse events， and Danggui Niantongtang had the highest probability of adverse reactions during treatment. According to the results of cluster analysis， Mahuang Lianyao Chixiaodou Tang and Simiaowan are effective and safe. ConclusionAccording to the results of NMA， Chinese medicinal prescriptions can assist in the treatment of AGA and improve the effectiveness of western medicine. For patients with AGA， clinicians can choose Mahuang Lianyao Chixiaodou Tang or Simiaowan as an auxiliary drug for routine western medicine treatment.
ABSTRACT
Objective To establish an artificial reflex arc to reinnervate the nerropathic bladder and restore bladder function after spinal cord injury. It involves a somatic reflex are with a modified efferent branch which passes the somatic motor impulses to the bladder and initiate autonomic bladder detrusor contraction. Methods This is achieved by intradural microanastomosis of the right L5 ventral root to S2 ventral root in canines. After a-conal regeneration,the pateller ligament-spinal cord center-bladder new artificial bladder reflex pathway was extablished. The early and final function of the reflex are was observed by electrophysiological experiments, bladder pressure tests and detrusor electromyograms (EMG)' Surgical operation of bilateral intradural microanastomosis of the L5 ventral roots to the S2 ventral roots was done in a 20 years old male patient with upper thoracispinal paraplegia. Results Single stmuli (115 mV, 1. 0 ms) of the right L, dorsal root resulted in evoked potentials recorded from the right S2 ventral root distal to the anastomosized site in all 6 canines. In 4 canines 6 months postoperation and 48 hours after spinal cord was transected, bladder contraction was very quickly initiated by trains of stimuli (l 000 mV, 10 Hz, 2 s) of the right L5 dorsal root and bladder pressures increased rapidly to average 65% of normal compared to controls' Bladder contraction induced by knocking the right pateller ligament increased to average 51 % of controls througt the new reflex arc. In 2 canines 18 months postoperation,bladder pressures increased by the same stimuli to average 84% of controls and to average 62% of controls by knocking the pateller ligament. Stimuli (3. 8 mA, 1' 0 Hz) of the right L5 dorsal root and femoral nerve resulted in EMG recorded from the detrusor in 2 canines 18 months postoperation. The patient achieved controllable micturition by calcaneus tendon-spinal cord center-bladder new artificial bladder reflex pathway at 14 months postoperation. Micturition was very quickly initiated by passive dorsalflexion of either lateral ankle joint,and the results of bladder function was excellent at postoperation. Conclusion The somatic motor a-cons can regenerate into the parasympathetic endoneurial tubes of autonomic nerve. Using the survived somatic reflex under the horizon of spinal cord injury to reconstruct the bladder autonomic reflex arc by intradural microanasto- mosis of ventral root is successful in the first patient and have a potential of clinical application.