[摘 要] 目的：探讨高良姜素（Gal）调控AMPK/mTOR/ULK1信号通路对胃癌细胞凋亡和自噬的影响及其机制。方法：将胃癌NCI-N87细胞分为对照组、多索吗啡（DM）组、Gal低剂量（Gal-L）组、Gal高剂量（Gal-H）组、Gal-H + DM组。采用MTT法、流式细胞术、划痕愈合实验和Transwell实验分别检测各组细胞的增殖、凋亡、迁移和侵袭能力，WB法检测PCNA、C-caspase-3、免疫逃逸相关蛋白（B7H1）、EMT和AMPK/mTOR/ULK1信号通路蛋白的表达水平。建立裸鼠NCI-N87细胞移植瘤模型，观察Gal和5-FU对移植瘤的抑制效果。结果：与对照组比较，DM组NCI-N87细胞增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著升高（均P < 0.05），细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著降低（均P < 0.05）；Gal-L组和Gal-H组NCI-N87细胞的增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著降低（均P < 0.05），细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著升高（均P < 0.05）；DM可部分逆转Gal对NCI-N87细胞恶性生物学行为的抑制作用（P < 0.05）；与对照组比较，Gal组和5-FU组裸鼠移植瘤体积和质量均显著降低，肿瘤组织细胞凋亡率显著升高（P < 0.05）。结论：Gal可促进胃癌NCI-N87细胞自噬和凋亡，抑制其增殖、迁移和侵袭，可能与激活AMPK/mTOR/ULK1信号通路有关。

Objective To investigate the effect of FUNDC1 tyrosine phosphorylation site mutations on mitophagy in H9c2 myocardial cells by constructing tyrosine site mutant plasmids (Y11 and Y18) of the FUN14 domain-containing protein 1 (FUNDC1). Methods The mutant plasmids constructed by whole-gene synthesis were transfected into rat myocardial H9c2 cells and divided into five groups: empty plasmid group, FUNDC1 overexpression group, Y11 mutant group, Y18 mutant group, and Y11 combined with Y18 mutant group. The viability of H9c2 cells was assessed using the CCK-8 assay. Additionally, tetramethylrhodamine ethyl ester (TMRE) staining was utilized to detect mitochondrial membrane potential. The protein expression levels of FUNDC1, translocase of the outer mitochondrial membrane 20 (TOM20), and cytochrome c oxidase IV (COX IV) were detected by Western blot analysis. Confocal microscopy was used to evaluate transfection efficiency as well as the co-localization of mitochondria and lysosomes. Results The FUNDC1 overexpression, Y11, Y18, and Y11 combined with Y18 mutant plasmids were successfully constructed. After plasmid transfection, widespread GFP fluorescence expression was observed under confocal microscopy. Compared with the empty plasmid group, FUNDC1 protein expression levels were significantly increased in the FUNDC1 overexpression group, Y11 mutation group, Y18 mutation group, and Y11 combined with Y18 mutation group, while cell viability and mitochondrial membrane potential showed no significant changes. Compared to the empty plasmid group, cells transfected with Y18 and Y11 combined with Y18 mutant plasmids showed increased TOM20 and COX IV expression levels and decreased mitochondrial-lysosomal co-localization. Conclusion Transfection with FUNDC1 Y18 or Y11 combined with Y18 mutant plasmids inhibited mitophagy in H9c2 myocardial cells.
Animals ; Rats ; Mitophagy/genetics* ; Myocytes, Cardiac/cytology* ; Mitochondrial Proteins/metabolism* ; Mutation ; Phosphorylation ; Tyrosine/genetics* ; Cell Line ; Membrane Proteins/metabolism* ; Membrane Potential, Mitochondrial

This consensus will introduce the characteristics of fillers used in the surgical cavities of domestic nasal surgery patients based on relevant literature and expert opinions. It will also provide recommendations for the selection of cavity fillers for different nasal diseases, with chronic sinusitis as a representative example.
Humans ; Nasal Cavity/surgery* ; Nasal Surgical Procedures ; China ; Consensus ; Sinusitis/surgery* ; Dermal Fillers

Objective To screen the active chemical components with potential therapeutic effects against lung cancer in tea and to provide new insights into the treatment and prevention of lung cancer.Methods Based on net-work pharmacology,the main active components from 13 types of tea samples were analyzed using liquid chromatog-raphy-mass spectrometry(LC-MS).The targets of these small molecules were obtained from the BATMAN-TCM da-tabase to construct a"component-target-disease"network.Lung cancer-related disease targets were retrieved from the GeneCard and Malacard databases followed by Gene Ontology(GO)functional and KEGG pathway enrichment analyses of potential pharmacological targets.A protein-protein interaction(PPI)network was constructed using the STRING database.The molecular docking was employed to screen small molecules with potential anti-cancer ac-tivity,and their potential inhibition to proliferation of human non-small cell lung cancer cell line A549 and human large cell lung cancer cell line H460.Results A total of 37 active components and 429 targets were identified in tea,with 182 overlapping targets associated with lung cancer.GO analysis revealed that these targets were primarily involved in biological processes such as cell proliferation,response to stimuli,and metabolic processes.KEGG pathway analysis indicated that these targets were mainly enriched in the p53 signaling pathway,ErbB signaling pathway,and PI3K-Akt signaling pathway.PPI network analysis identified key targets including MAPK1,AKT1,SRC,MAPK3,and p53.Molecular docking screened coumestrol as a molecule capable of binding to human estro-gen receptor 2(ESR2),and its inhibitory effect on the proliferation of A549 and H460 cells was experimentally validated(P＜0.000 1).Conclusions The active components in tea may intervene in the development and progres-sion of lung cancer through a multi-component,multi-target,and multi-pathway mechanism,The results suggests po-tential components against lung cancer in tea,which may be applied in the prevention of human lung cancer.

Objective:To explore the differences in tumor-specific growth factors, cellular immune function and efficacy of olaparib and platinum-containing regimen for treatment of platinum-sensitive relapsed ovarian cancer patients with BRCA mutation.Methods:A retrospective cohort study was conducted. A total of 100 platinum-sensitive relapsed BRCA-mutant ovarian cancer patients in Baoding Second Central Hospital from September 2017 to March 2020 were retrospectively selected. The clinical data of the patients were analyzed, and they were divided into the olaparib group (treated with olaparib tablets) and the platinum-containing regimen group (treated with paclitaxel and platinum drugs for 6 cycles, followed by olaparib tablets maintenance therapy), with 50 patients in each group. The clinical efficacy, tumor specific growth factor [carbohydrate antigen (CA) 125, CA199, human epididymal protein 4 (HE4)] levels, cellular immune function-related indicators [T-cell subsets (proportions of CD3 + cells and CD4 + cells), CD4 + cells/CD8 + cells ratio (CD4 +/CD8 +)], and quality of life scores before treatment and after 2, 4 and 6 cycles of treatment of the two groups were compared, as well as the safety of the two groups. The data of three years of follow-up were obtained, Kaplan-Meier method was used to analyze the progression-free survival (PFS) of patients in the two groups, and log-rank test was used for comparison between groups. Results:The age of patients in the olaparib and platinum-containing regimen groups was (53±7) years old and (56±7) years old, respectively. The differences in compositions of patients with different age, body mass index, Eastern Cooperative Oncology Group (ECOG) performance status score, primary tumor location, lesion size, pathological stage, pathological type, germline BRCA mutation, and previous chemotherapy response between the two groups were not statistically significant (all P > 0.05). The objective response rate (ORR) [58.0% (29/50) vs. 38.0% (19/50)] and disease control rate (DCR) [80.0% (40/50) vs. 56.0% (28/50)] of the olaparib group after treatment were higher than those of the platinum-containing regimen group, and the differences were statistically significant (both P < 0.05). Serum CA125, CA199 and HE4 levels were gradually decreased in both groups before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05); serum CA125, CA199 and HE4 levels in the olaparib group after 2, 4 and 6 cycles of treatment were lower than those in the platinum-containing regimen group, and the differences were statistically significant (all P < 0.05). The CD3 + cells ratio, CD4 + cell ratio and CD4 +/CD8 + in the olaparib group gradually increased before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05), while those in the platinum-containing regimen group all gradually decreased (all P < 0.05); the CD3 + cells ratio, CD4 + cells ratio and CD4 +/CD8 + in the olaparib group were higher than those in the platinum-containing regimen group after 2, 4 and 6 cycles of treatment, and the differences were statistically significant (all P < 0.05). The quality of life scores of both groups increased before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05), and the quality of life scores of the olaparib group were higher than those of the platinum-containing regimen group after 2, 4 and 6 cycles of treatment, and the differences were statistically significant (all P < 0.05). The incidence of nausea, fatigue and malaise, vomiting, anemia, and diarrhea at all levels in the olaparib group was lower than those in the platinum-containing regimen group (all P < 0.05). By follow-up for 3 years, there was no statistically significant difference in PFS between the olaparib group and the platinum-containing regimen group ( P > 0.05). Conclusions:The efficacy of olaparib treatment in platinum-sensitive relapsed ovarian cancer patients with BRCA mutation is superior to platinum-containing regimen, and it can increase the level of T cells, inhibit the expression of tumor-specific growth factors, improve the quality of life, and have a positive effect on improving the safety of treatment.

Nitroxide radicals are a kind of stable organic free radicals.Due to the presence of N-O·and unpaired electrons in its structure,it has many characteristics,and thus can be used as a spin marker to explore the mechanism of biological reactions;with its magnetic properties,it can be used for the development of multifunctional magnetic molecular materials and used as a polymerization inhibitor and catalyst in organic reactions.More importantly,it has a variety of biological activities such as anti-oxidation and anti-tumor,and so has attracted much attention in the research and development of new drugs.For example,the spin labeling of nitroxide radicals on anticancer drug podophyllotoxin can enhance the efficacy and reduce the toxicity,and can be easily to be absorbed by the body,thus obtaining a new anti-cancer drug 4-[4″-(2″,2″,6″,6″-tetramethyl-1″-piperidinyloxy nitroxide radical)amino]-4′-demethyl epipodophyllotoxin(GP-7).It is an effective way to seek new drugs by introducing pharmacophore to modify nitroxide radicals or it can be spin-labeled on active natural products to obtain new compounds with high efficiency and low toxicity.The research progress of derivatives and its biological activitives of nitroxide radicals are summarized,aiming to provide theoretical basis for the developing and utilizing of nitroxide radicals and searching for new drugs.

Objective To explore the correlation between serum microRNA-29a-3p(miR-29a-3p),throm-bospondin 2(THBS2)and cardiopulmonary function in children with chronic pulmonary heart disease.Meth-ods A total of 136 children with chronic pulmonary heart disease treated in Shijiazhuang Hospital of Tradi-tional Chinese Medicine from July 2019 to September 2023 were selected as the study subjects.Based on their clinical signs,cardiopulmonary function,and symptoms,they were divided into a compensated group(74 ca-ses)and a decompensated group(62 cases).Real-time fluorescence quantitative PCR(RT-qPCR)method was applied to detect serum miR-29a-3p level,and enzyme-linked immunosorbent assay(ELISA)was applied to detect serum THBS2 level.Moreover,ultrasound diagnostic equipment was applied to detect cardiac function indicators such as left ventricular ejection fraction(LVEF)and cardiac output(CO).Creatine kinase isoenzyme(CK-MB)and troponin(cTnl)were detected by electroluminescence analysis.Pulmonary function indicators were detected by pulmonary artery systolic pressure(PASP),mean pulmonary artery pressure(MPAP)and pulmonary artery diastolic pressure(PADP).Pearson correlation was used to analyze the corre-lation between serum miR-29a-3p and THBS2 levels and cardiopulmonary function indexes in children with chronic pulmonary heart disease,and multivariate Logistic regression analysis was used to screen the influen-cing factors of the disease in children with chronic pulmonary heart disease.Receiver operating characteristic(ROC)curve was drawn to analyze the evaluation value of serum miR-29a-3p and THBS2 levels in children with chronic pulmonary heart disease.Results The THBS2,CK-MB,cTnI,PASP,MPAP,and PADP in the decompensated group were higher than those in the compensated group(P＜0.05),while miR-29a-3p,LVEF,and CO were lower than those in the compensated group(P＜0.05).Serum miR-29a-3p in children with chro-nic pulmonary heart disease was positively correlated with LVEF and CO(P＜0.05),but negatively correla-ted with CK-MB,cTnI,PASP,MPAP,and PADP(P＜0.05).THBS2 was negatively correlated with LVEF and CO(P＜0.05),but positively correlated with CK-MB,cTnI,PASP,MPAP,and PADP(P＜0.05).MiR-29a-3p was a protective factor for exacerbation of chronic pulmonary heart disease in children(P＜0.05),while THBS2 was an independent risk factor for exacerbation of chronic pulmonary heart disease in children(P＜0.05).ROC curve analysis results showed that the area under the curve(AUC)of serum miR-29a-3p and THBS2 levels alone and in combination for evaluating the severity of chronic pulmonary heart disease in children were 0.827,0.817 and 0.915,respectively.Conclusion Serum miR-29a-3p and THBS2 levels are both factors affecting the disease of children with chronic pulmonary heart disease,and are closely related to the disease and cardiopulmonary function of children with chronic pulmonary heart disease.

Objective:To evaluate the effect of the right stellate ganglion block (SGB) on the acute lung injury induced by hind limb ischemia-reperfusion (I/R) in rats.Methods:Twenty-seven SPF healthy male Sprague-Dawley rats, aged 7-8 weeks, weighing 200-250 g, were divided into 3 groups ( n=9 each) by the random number table method: sham operation group (Sham group), hind limb I/R group (I/R group) and right SGB+ hind limb I/R group (SGB+ I/R group). The right hind limb I/R injury model was developed in anesthetized rats in I/R and SGB+ I/R groups, and the right SGB was performed with 0.2% ropivacaine 0.5 ml before the right hind limb ischemia in SGB+ I/R group. The pathological changes of lung tissues were observed with a light microscope, the wet/dry lung weight ratio, PaO 2 and PaCO 2 were determined, and the oxygenation index was calculated. The concentrations of serum malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) were determined using enzyme-linked immunosorbent assay. Results:Compared with Sham group, the wet/dry lung weight ratio, PaO 2, oxygenation index, and GSH-Px concentrations were significantly decreased, and the concentrations of serum MDA were increased in I/R and SGB+ I/R groups ( P<0.05). Compared with I/R group, the wet/dry lung weight ratio, PaO 2, oxygenation index, and GSH-Px concentrations were significantly increased, the concentrations of serum MDA were decreased ( P<0.05), and the pathological changes of lung tissues were significantly attenuated in SGB+ I/R group. There was no significant difference in PaCO 2 among the 3 groups ( P>0.05). Conclusions:The right SGB can alleviate the acute lung injury induced by hind limb I/R, and the mechanism may be related to the inhibition of oxidative stress responses in rats.

ObjectiveTo perform a predictive analysis of the quality marker（Q-Marker） for the anticoagulant activity of Kunning granules. MethodThe chemical components of Kunning granules were analyzed by ultra high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry（UHPLC-Q-TOF-MS/MS） on a Waters ACQUITY UPLC HSS T3 column（2.1 mm×100 mm， 1.8 μm） with the mobile phase of acetonitrile（A）-25 mmol∙L^-1 ammonium acetate aqueous solution（B） for gradient elution （0-5 min， 5%-22%A； 5-10 min， 22%-30%A； 10-15 min， 30%-95%A； 15-20 min， 95%-5%A； 20-30 min， 5%A）， flow rate of 0.2 mL∙min^-1， column temperature at 30 ℃， injection volume of 1 μL， electrospray ionization（ESI）， positive and negative ion detection modes. Interaction analysis between the targets of chemical components and the targets of abnormal uterine bleeding（AUB） was performed by network pharmacology， and the key components were screened through network topology analysis. The fingerprints of 10 batches of Kunning granules were established by high performance liquid chromatography（HPLC）， the anticoagulant activity of the granules was determined by blood coagulation method and fibrinogen plate method， and the spectrum-effective relationship was established. The components co-occurring in the topological analysis and spectrum-effective relationship were selected as Q-Markers， and their anticoagulant activities were verified and confirmed. ResultA total of 475 chemical components were identified from Kunning Granule， of which 22 key components such as salvianolic acid B， paeoniflorin， naringin and neohesperidin， were the potential material basis for the treatment of AUB. The spectrum-effective analysis showed that peaks 7（paeoniflorin）， 9（naringin）， 10（neohesperidin） and 11（salvianolic acid B） were the optimal principal components， and in vitro activity test showed that these four components could better characterize their anticoagulant activity. ConclusionSalvianolic acid B， paeoniflorin， neohesperidin and naringin may be Q-Markers for the anticoagulant activity of Kunning granules.

Lymphangioleiomyomatosis (LAM) is a rare, multisystemic, low-grade neoplasm character-ized by diffuse cystic lesions in the lung.In recent years, emerging imaging examination such as ⁶⁸Ga-NEB PET-CT scan provides efficient and precise non-invasive diagnostic methods to detect lymphatic circulation abnormalities in LAM patients. The long-term efficacy and safety of sirolimus for LAM has accumulated further evidence, and genetic profiling studies have unveiled more information of genetic mechanisms. Prognosis of LAM has been much improved. We briefly reviewed the research advances of LAM in China and other countires.