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ObjectiveTo predict the potential targets and possible related signaling pathways of Salviae Miltiorrhizae Radix et Rhizoma against bladder cancer （BC） based on network pharmacology and verify the potential molecular mechanism through in vitro cell experiment. MethodActive components of Salviae Miltiorrhizae Radix et Rhizoma were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and BC-related targets were searched from GeneCards and Online Mendelian Inheritance in Man （OMIM）. Via Venny2.1， the potential targets of Salviae Miltiorrhizae Radix et Rhizoma against BC were screened out and the Venn diagram was plotted. Protein-protein interaction （PPI） network was constructed by STRING， followed by Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Gnomes （KEGG） pathway enrichment with DAVID. Cell Counting Kit-8 （CCK-8） assay was employed to detect the inhibitory effect of tanshinone ⅡA （Tan ⅡA）， cryptotanshinone （CPT）， and luteolin （LUT） at different concentration （0， 1， 2， 4， 8， 16， 32 μmol·L-1） on the proliferation of BC T24 and 5637 cells， propidium iodide （PI） staining to analyze the apoptosis of 5637 cells induced by Tan ⅡA， CPT， and LUT （0， 4， 8 μmol·L-1）， and Western blotting to detect the regulatory effect of Tan ⅡA （0， 4， 8， 16 μmol·L-1） on the expression of key target proteins. ResultA total of 65 active components and 39 anti-BC targets of Salviae Miltiorrhizae Radix et Rhizoma were screened out. The anti-BC targets were mainly involved in the KEGG pathways of neuron-ligand-receptor interaction， phosphatidylinositol 3-kinases （PI3K）/protein kinase B （Akt） signaling pathway， epidermal growth factor receptor （EGFR） tyrosine kinase inhibitor resistance， and hypoxia inducible factor （HIF）-1 signaling pathway. As for the CCK-8 assay， compared with the blank group， Tan ⅡA， CPT， and LUT significantly inhibited the proliferation of T24 and 5637 cells， particularly the 5637 cells. The half maximal inhibitory concentration （IC50） of Tan ⅡA on 5637 cells was significantly lower than that of CPT and LUT. Moreover， compared with the blank group， Tan ⅡA， CPT， and LUT all induced the apoptosis of 5637 cells， and the effect followed the order of Tan ⅡA>CPT>LUT （P<0.05）. Western blot showed that Tan ⅡA significantly reduced the expression of EGFR， p-PI3K， and p-Akt in 5637 cells in a concentration-dependent manner compared with the blank group （P<0.05）. ConclusionSalviae Miltiorrhizae Radix et Rhizoma exerts therapeutic effect on BC through multiple components， multiple targets， and multiple pathways. The mechanism is the likelihood that it down-regulates the expression of EGFR， p-PI3K， and p-Akt proteins， thus further inhibits cell proliferation， and induces apoptosis.
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The ban on addition of antibiotics in animal feed in China has made the search for new antibiotics substitutes, e.g. bacteriocin, a hot topic in research. The present study successfully isolated an antibacterial substance producing strain of Bacillus sp. from alpaca feces by agar diffusion method, using Escherichia coli, Salmonella enterica, Staphylococcus aureus, Staphylococcus epidermidis, Micrococcus luteus and Listeria monocytogenes as indicator bacteria. The isolated strain was named as B. licheniformis SXAU06 based on colony morphology, Gram staining and 16S rRNA gene sequence. The antibacterial substance was isolated and purified through a series of procedures including (NH4)2SO4 precipitation, chloroform extraction, molecular interception and SDS-PAGE analysis. Bioinformatics analysis of the LC-MS/MS data indicated that the antibacterial substance was a bacteriocin-like substance (BLIS) with an approximate molecular weight of 14 kDa, and it was designated as BLIS_SXAU06. BLIS_SXAU06 exhibited high resistance to treatment of proteinase K, high temperature, high acidity and alkalinity. BLIS_SXAU06 was heterologously expressed in E. coli and the recombinant BLIS_SXAU06 exhibited effective antibacterial activity against S. aureus, S. epidermidis, M. luteus, and L. monocytogenes, showing potential to be investigated further.
Subject(s)
Animals , Anti-Bacterial Agents/pharmacology , Bacillus licheniformis , Bacteriocins/pharmacology , China , Chromatography, Liquid , Escherichia coli/genetics , Listeria monocytogenes , RNA, Ribosomal, 16S , Staphylococcus aureus , Tandem Mass SpectrometryABSTRACT
Lung cancer, which is exacerbated by environmental pollution and tobacco use, has become the most common cause of cancer-related deaths worldwide, with a five-year overall survival rate of only 19% (Siegel et al., 2020; Yang et al., 2020; Yu and Li, 2020). Nearly 85% of lung cancers are non-small cell lung cancers, of which lung adenocarcinoma is the most common subtype accounting for 50% of non-small cell lung cancer cases. At present, radiotherapy is the primary therapeutic modality for lung cancer at different stages, with significant prolongation of survival time (Hirsch et al., 2017; Bai et al., 2019; Shi et al., 2020). Irradiation can generate reactive oxygen species (ROS) through the radiolysis reaction of water and oxygen, cause DNA damage and oxidative stress, and subsequently result in cancer cell death (Kim et al., 2019). Nevertheless, radioresistance seriously hinders the success of treatment for lung cancer, owing to local recurrence and distant metastasis (Huang et al., 2021). Compared with small cell lung cancer, non-small cell lung cancer shows more tolerance to radiotherapy. Therefore, it is of great importance to decipher key mechanisms of radioresistance and identify effective molecular radiosensitizers to improve patient survival.
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This article was aimed to study the relevance on traditional Chinese medicine (TCM) syndromes and TNM staging and tumor antigen in primary lung cancer, in order to provide theoretical supports for TCM prevention and treatment as well as medication guidance of primary lung cancer. Statistical data from 388 lung cancer patients were analyzed according to the relevance TCM syndromes, in order to find the relation between syndrome distribution of lung cancer and TNM staging and tumor antigens. The results showed that in TNM staging, the incidence of lung-yin deficiency syndrome in I stage was apparently higher than that in IV stage; the incidence of spleen-qi deficiency syndrome in IV stage was apparently higher than that in I stage; and there was no obvious difference in lung-qi deficiency syndrome, stasis obstructing lung collateral syndrome or lung-yin deficiency with fire-excess. The proportion of CEA in lung-yin deficiency with fire-excess syndrome was significantly higher than that in lung-yin deficiency syndrome, lung-qi deficiency syndrome, and stasis obstructing lung collateral syndrome. The proportion of abnormal increasing of NSE in lung-yin deficiency with fire-excess syndrome was significantly higher than other syndromes. The proportion of abnormal increasing of CYFRA21-1 in lung-yin deficiency with fire-excess syndrome and stasis obstructing lung collateral syndrome was significantly higher than that in lung-qi deficiency syndrome. It was concluded that there were certain relevance between TCM syndromes and TNM staging in lung cancer. Lung-yin deficiency syndrome, which existed in all stages of lung cancer, was the most obvious in the early stage. Spleen-qi deficiency syndrome was commonly seen in the advanced stage of lung cancer. There were certain relevance between different TCM syndromes and the abnormal increasing of tumor antigens including CEA, NSE and CYFRA21-1.
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Objective HGF and its receptor ( c-met) are principal mediators of mesenchymal-epithelial interac-tion in several different systems.Moreover, hair follicle is a model of mesenchymal-epithelial interaction.The aim of this study was to explore whether HGF/c-met signal plays a role in the control of hair follicle growth cycle.Methods To ex-amine the localization of HGF/c-met in anagen, catagen and telogen in the hair follicle of ICR mice.Results HGF was mainly located in the dermal papilla and sebaceous gland, and c-Met in the hair matrix, root sheath and epidermis.Both HGF and c-met expressions peaked during anagen, not found in catagen, and increased during telogen-anagen phase.Con-clusions Our results demonstrate a regulatory role of HGF and c-met in the control of hair follicle growth in ICR mice.
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To study the effect of Feiyanning Decoction (FYN), a compound traditional Chinese medicine, on expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by tumor necrosis factor-α (TNF-α) in human lung adenocarcinoma epithelial cell line (A549).
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To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on the ratio of CD4(+)CD25(+) regulatory T cells and expression of transcription factor Foxp3 in mice with Lewis lung cancer.
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Objective To study the protective effect of octreotide on myocardial injury after hepatic ischemia-reperfusion in rabbit. Methods Pringle's maneuver rabbit hepatic ischemia-reperfusion model was established. 24 adult New Zealand rabbits were random divided into equal 3 groups: sham operative group ( group A) , ischemia-reperfusion group( group B) and octreotide preconditioning group ( group C ). The levels of CK-MB( MB isoenzyme of creatine kinase) and LDH ( 1actate dehydrogenase), superoxide dismutase (SOD) and malondialdehyde (MDA) of each group were measured at the time before ischemia (T1) , after ischemia for 30 mins ( T2 ) and after reperfusion for 60 mins ( T3 ), 120mins ( T4 ), 240 mins ( T5 ). The SOD and MDA in myocardial tissue of each group were measured after reperfusion for 240 mins. The changes of ultrastructure in the myocardial cell were observed by transmission electron microscopy after reperfusion for 240 mins. Results There was no significant difference in the levels of CK-MB and LDH in serum of each group before ischemia ( P ＞0. 05). The CK-MB and LDH of group B and C were higher than that of group A ( P ＜0.05) after ischemia for30 mins. The CK-MB and LDH of group C were lower than that of group B in this period( P ＜0. 05 ). The highest time point of LDH and CK-MB were after reperfusion for 120 mins and 240 mins. The contents of MDA in group B and group C were higher than that in group A from after ischemia for 30 mins in plasma and after reperfusion for 240 mins in myocardial tissue ( P ＜ 0. 05 ),and it in group C were lower than that in group B( P ＜0.05) .The contents of SOD in group B and group C were lower than that in group A from after ischemia for 30 mins in blood plasma and after reperfusion for 240 mins in myocardial tissue ( P ＜0. 05), and in group C were higher than that in group B( P ＜0. 05).The electromicroscope showed that the pathological change of myocardial ultrastructure of group C was slighter than that of group B. Conclusion Octreotide can stabilize myocardial cell membrane and reduce release of oxygen free radical and significantly relieve the injury of myocardial ultrastructure after hepatic ischemiareperfusion in rabbit. Octreotide preconditioning can relieve myocardial injury after hepatic ischemia-reperfusion in rabbit.
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Objective To evaluate the blond-saving effect of low central venous pressure(CVP) combined with acute hypervolemic hemedilution(AHHD)in patients undergoing hepatic lobectomy.Methods sixty ASA I orⅡpatients of both sexes aged 32-48 yr weighing 47-72 kg undergoing hepatic lobectomy for primary malignant hepatonm under epidural combined with general anesthesia were randomly divided into 3 groups(n=20 each);group I control(C);group 1I AHHD and group Ⅲ low CVP+AHHD.Group C received crystalloid and coloid in a ratio of 1.5:1 during operation.In groupⅡ4% suecinylated gelatin was infused at 50 ml·kg-1·h-1 for 30 min after tracheal intubation (AHHD);while inⅢ group low CVP was induced and maintained by epidural administration of a mixture of 1.5% lidnoaine +O.2% bupivacaine 6-8 ml combined with intravenous infmion of propofol at 6 mg·kg-1·h-1 until 10 min after hepatic lobectomy was completed.then 4% succinylated gelatin was infused at 50 ml·kg-1·h-1 for 30 min.Blood glucose,Hb,Hct, WBC count,blood coagulation (PT,AVIT,Fib),shtmic-pyruvic transaminase (GPT) and renal function (BUN,Cr) were determined before operation (baseline),immediately before skin incision,immediately before and 10 min after liver lobe was removed,at the end of operation and 7 d after operation.Urine output,intraoperative blood loss and blood transfusion and complications were recorded.Results The glood glucose concentration.WBC count and GPT levd were significantly lower;the amount of fluid infused and urinary output before hepatic lobe resection and the percentage of the patients with allogeneic blood transfusion during operation were less;Hb,Hct and the amounl of fluid infused and urinary output after hepatic lobe resection were uigher in grolp Ⅲ than in group I and ⅡⅡⅡ.There were no significant differences in blood coagulation,renal function,the total amount of fluid infused and urine output among the 3 groups.No patient developed any complication.Conclusion The low CVP hefor combined with AHHD after hepatic resection can decrease intraoperative blood loss and allogeneic blood transfusior and is safe.
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Objective To observe transforming growth factor-?1(TGF-?1) in chicken callus. Methods The right humerus fracture models of 16 New Roman hens, at the age of 6-month old, about 1.75kg in weight, were established under no germ operational condition. TGF-?1 of the tissue samples procured from callus on 3d,6d,10d and 15d of fracture healing were examined by immunohistochemical method. Results Histochemical study shows that the immunostaining of TGF-?1 exists in blood cells on 3d, connective tissue cells and extracellular matrix on 6d, cartilage on 10d and 15d. Compared with that on 3d, the expression of TGF-?1 increased markedly on 6d, 10d and 15d of fracture healing(P
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Objective To study the expression and localization of TGF?1 and its signal transducer Smad2 and Smad4 which play important roles during testicular development and spermatogenesis in alpaca testis. Methods The whole testes were obtained from alpacas aged 24 months(n=3) at the Scientific Research Base of Shanxi Agriculture University.The protein expressions of TGF-?1、smad2、smad4 in alpaca testes were examined by Western blotting and SABC. Results The distributions of(TGF-?1)、Smad2 and Smad4 in the testes of alpacas aged 24 months were demonstrated.Conclusion Our findings show that TGF-?1、Smad2 and Smad4 are involved in the regulation of spermatogenesis,and provide direct evidences for the moleculer mechnism of TGF-?1 action in the alpaca testis during spermatogenesis.
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CB-HRP was injected into the ovary of the domestic hens of 75-90 days old to trace the originating neurons of the vagus nerve innervated the ovary. The results were as the following:1. The afferent vagus neurons innervated the ovary were located in the nodose ganglia and the jugular ganglia. The afferent fibers in the ovarian medulla were found chiefly in the solitary tract, the nucleus of the solitary tract and the commissural nucleus of Cajal also.2. The efferent vagus neurons were located mainly in the subnueleus ventralis parvicellularis (VP)and the subnueleus ventrolateralis (VL)of the dorsal vagal motor nucleus, and a small number of neurons extended from the subnueleus VP and VL to the neighbouring five subnuclei.