Résumé
Network pharmacology, molecular docking, and in vivo and in vitro experiments were employed to study the molecular mechanism of Blaps rynchopetera Fairmaire in the treatment of non-small cell lung cancer(NSCLC). The components of B. rynchopetera were collected by literature review, and the active components were screened out through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). PharmMapper was used to obtain the targets of the active components. The targets of NSCLC were obtained from DrugBank, GeneCards, OMIM, TTD, and PharmGKB. The Venn diagram was drawn to identify the common targets shared by the active components of B. rynchopetera and NSCLC. The "drug component-target" network and protein-protein interaction(PPI) network were constructed by Cytoscape, and the key targets were screened by Centiscape. Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment of the above key targets were performed by DAVID. AutoDock and PyMOL were used for the molecular docking between the key targets and corresponding active components. A total of 31 active components, 72 potential targets, and 11 key targets of B. rynchopetera against NSCLC were obtained. The active components of B. rynchopetera had good binding activity with key targets. Further, the serum containing B. rynchopetera was prepared and used to culture human lung adenocarcinoma A549 cells. The CCK-8 assay was employed to determine the inhibition rates on the growth of A549 cells in blank control group and those exposed to different concentrations of B. rynchopetera-containing serum, cisplatin, and drug combination(B. rynchopetera-containing serum+cisplatin) for different time periods. The cell migration and invasion of A549 cells were detected by cell scratch assay and Transwell assay, respectively. Western blot was employed to determine the expression levels of B-cell lymphoma-2(Bcl-2), Bcl-2-associated X(Bax), caspase-3, cell division cycle 42(CDC42), proto-oncogene tyrosine-protein kinase SRC, and vascular endothelial growth factor(VEGF) in A549 cells. C57BL/6 mice were inoculated with Lewis cells and randomly assigned into a model control group, a B. rynchopetera group, a cisplatin group, and a drug combination(B. rynchopetera+cisplatin) group, with 12 mice per group. The body weight and the long diameter(a) and short diameter(b) of the tumor were monitored every other day during treatment, and the tumor volume(mm~3) was calculated as 0.52ab~2. After 14 days of continuous medication, the mice were sacrificed for the collection of tumor, spleen, and thymus, and the tumor inhibition rate and immune organ indexes were calculated. The tissue morphology of tumors was observed by hematoxylin-eosin(HE) staining, and the positive expression of Bax, Bcl-2, caspase-3, CDC42, SRC, and VEGF in the tumor tissue was detected by immunohistochemistry. The results indicated that B. rynchopetera and the drug combination regulated the expression levels of Bax, Bcl-2, caspase-3, CDC42, SRC, and VEGF to inhibit the proliferation, migration, and invasion of A549 cells and Lewis cells, thus playing a role in the treatment of NSCLC via multiple ways.
Sujets)
Humains , Animaux , Souris , Souris de lignée C57BL , Carcinome pulmonaire non à petites cellules/génétique , Caspase-3 , Pharmacologie des réseaux , Facteur de croissance endothéliale vasculaire de type A , Cisplatine , Simulation de docking moléculaire , Protéine Bax , Tumeurs du poumon/génétique , Prolifération cellulaire , Médicaments issus de plantes chinoises/pharmacologie , Médecine traditionnelle chinoiseRésumé
This study aims to investigate the effects of Blaps rynchopetera Fairmaire and/or cyclophosphamide on the proliferation and apoptosis of lung cancer cells and decipher the underlying mechanism. B. rynchopetera and cyclophosphamide-containing serum and blank serum were prepared from SD rats. Cell counting kit-8(CCK-8) assay was employed to examine the proliferation of lung cancer cell lines A549 and Lewis treated with corresponding agents. The Jin's formula method was used to evaluate the combined effect of the two drugs. According to the evaluation results, appropriate drug concentrations and lung cancer cell line were selected for subsequent experiments, which included control, B. rynchopetera, cyclophosphamide, B. rynchopetera + cyclophosphamide, and B. rynchopetera + Wnt/β-catenin pathway agonist lithium chloride(LiCl) groups. Immunocytochemistry was employed to measure the expression of proliferation-related proteins in Lewis cells after drug interventions. Flow cytometry was employed to determine the cell cycle and apoptosis. The expression levels of proliferating cell nuclear antigen(PCNA), cyclinD1, B-cell lymphoma 2(Bcl-2), Bcl-2-assiocated X protein(Bax), Wnt1, and β-catenin were determined by Western blot. The results showed that B. rynchopetera and/or cyclophosphamide significantly inhibited the proliferation of A549 and Lewis cells. Compared with B. rynchopetera alone, the combination increased the inhibition rate on cell proliferation. The combination of B. rynchopetera and cyclophosphamide demonstrated a synergistic effect according to Jin's formula-based evaluation. Compared with the control group, the B. rynchopetera, cyclophosphamide, and B. rynchopetera + cyclophosphamide groups showed increased proportion of Lewis cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the cyclophosphamide group, the combination group showed increased proportion of cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the B. rynchopetera group, the B. rynchopetera + LiCl group had deceased proportion of cells in G_0/G_1 phase, decreased apoptosis rate, down-regulated expression of Bax, and up-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. The results indicated that B. rynchopetera could inhibit the proliferation, arrest the cell cycle, and induce the apoptosis of lung cancer cells by inhibiting the Wnt/β-catenin signaling pathway. Moreover, B. rynchopetera had a synergistic effect with cyclophosphamide.
Sujets)
Rats , Animaux , Voie de signalisation Wnt , Tumeurs du poumon/génétique , bêta-Caténine/métabolisme , Antigène nucléaire de prolifération cellulaire , Protéine Bax/métabolisme , Rats de lignée LEW , Rat Sprague-Dawley , Apoptose , Protéines proto-oncogènes c-bcl-2/métabolisme , Prolifération cellulaire , Cyclophosphamide , Lignée cellulaire tumoraleRésumé
OBJECTIVE@#To observe the effect of acupuncture combined with regular treatment and swallowing function training on pharyngeal motor, sensory function and penetration-aspiration function in patients with dysphagia after stroke.@*METHODS@#A total of 60 patients with dysphagia after stroke were randomly divided into a control group and an observation group, 30 patients in each group. Both groups were treated with conventional treatment and swallowing function training; in addition, the observation group was treated with acupuncture at Lianquan (CV 23), Fengfu (GV 16), Yifeng (TE 17). All the treatments were given once a day, 5 days a week, for totally 4 weeks. In the two groups, the pharyngeal motor and sensory function, penetration-aspiration scores were evaluated by fiberoptic endoscopic evaluation of swallowing (FEES), and the Kubota water swallowing test scores were assessed before and after treatment, and the clinical effects were compared.@*RESULTS@#After treatment, the pharyngeal motor and sensory function in the two groups were all higher than those before treatment (P<0.05), and those in the observation group were better than the control group (P<0.05). After treatment, the penetration-aspiration scores and Kubota water swallowing test scores in the two groups were all lower than those before treatment (P<0.05), and those in the observation group were lower than the control group (P<0.05). The total effective rate was 93.3% (28/30) in the observation group, which was better than 73.3% (22/30) in the control group (P<0.05).@*CONCLUSION@#Acupuncture combined with regular treatment and swallowing training could improve the pharyngeal motor and sensory function, and penetration-aspiration scores in patients with dysphagia after stroke.
Sujets)
Humains , Points d'acupuncture , Thérapie par acupuncture , Déglutition , Troubles de la déglutition/thérapie , Accident vasculaire cérébral/complications , Résultat thérapeutique , EauRésumé
Objective To evaluate the effect of personnel activities and air purifiers on airborne microorganisms and particulate matter in bronchoscopy room.Methods According to whether there was personal activity and air purifier in the bronchoscopy room,the experiment was divided into four groups:dynamic non-purification group,dynamic purification group,static non-purification group,and static purification group,indoor air samples were collected and analyzed at five different time points (0,0.5,1,2,4 h),microorganisms in the air were collected by planktonic method,then cultured and counted,concentration of particulate matter was determined by DT-9881M laser dust particle counter,variance analysis of factorial design was used for statistical analysis.Results Colony count/concentration of airborne bacteria,fungi,total microorganisms (bacteria + fungi),PM2.5,and PM2.5-10.0 in dynamic non purification group were (113.53 ± 7.78) CFU/m3,(89.67 ± 7.17) CFU/m3,(203.20 ± 10.92) CFU/m3,(86 557.20 ±4 158.29) counts/m3,and (659.69 ± 38.91) counts/m3 respectively,in static non-purification group were (84.33 ± 3.65) CFU/m3,(65.00 ± 2.65)CFU/m3,(149.33 ± 4.98) CFU/m3,(45 812.64 ±1 279.61) counts/m3,and (189.15 ± 4.64) counts/m3 respectively,in dynamic purification group were (84.80 ±8.08) CFU/m3,(90.40 ± 5.50) CFU/m3,(175.20 ± 9.22) CFU/m3,(49 336.38 ± 2 039.16) counts/m3,and (218.36 ± 7.02) counts/m3 respectively,in static purification group were (67.80 ± 5.63) CFU/m3,(38.27 ± 3.70)CFU/m3,(106.07 ± 6.76) CFU/m3,(29 772.53 ± 2 212.93) counts/m3,and (124.80 ± 7.16) counts/m3 respectively.Colony count/concentration of airborne bacteria,total microorganisms,PM2.5,and PM2.s 10.0 in dynamic group were all higher than those in static group,non-purification group were higher than purification group(both P <0.05),colony count of fungi in dynamic non-purification group was higher than static non-purification group,in static purification group was lower than static non-purification group(both P<0.05),there was no significant difference between dynamic purification group and dynamic non-purification group (P =0.936).Conclusion Personal activities can increase colony count/concentration of microorganisms and particulate matter in bronchoscopy room,air purifier can reduce the bacteria,total microbial count,and particulate matter in the air of bronchoscopy room.
Résumé
Objective To study the clinical and genetic profile of the patients with Gitelman syndrome ( GS ) . Methods We retrospectively analyzed the gene mutation and clinical character of 64 GS patients diagnosed in Pe-king Union Medical College Hospital from 2012 to 2016 .Results The age at diagnosis of these 64 patients ( 39 male,25 female) were (35±14) years.At admission, the serum potassium level of the patients was (2.86± 0.44) mmol/L, serum magnesium level was ( 0.62 ±0.14 ) mmol/L, 24-hour urine potassium was ( 82.27 ± 39.73)mmol/day, 24-hour urine calcium was (0.94±0.83)mmol/day and their average blood pressure was 110/69 mmHg.The gene mutation were divided into four groups including homozygous mutation ( n=5) , compound het-erozygous mutation(n=40), multiple mutations (n=9) and single heterozygous mutation (n=10) group.The compound heterozygous group had higher serum potassium concentration ( P<0.05 ) and the homozygous group had a relatively higher serum magnesium concentration but without significance .A total of 74 distinctly different mutation alleles were identified , of which 24 were new mutation alleles .p.Asp486Asn was a hotspot in our series which was found in 16 patients ( 25.0%) .Conclusions There exists great heterogeneity of gene mutation and clini-cal character in Gitelman syndrome .Patients with compound heterozygous have a relatively milder clinical character.p.Asp486Asn mutation is a hotspot in Chinese patients .