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Objective To investigate the role of microRNA(miR)-567 in the proliferation,migration and cell cycle of non-small cell lung cancer(NSCLC)through regulation of cyclin dependent kinase 8(CDK8)and its clinical relevance.Methods Tumor tissues and adjacent tissues of 40 NSCLC patients were collected,and the expressions of miR-567 and CDK8 were detected by real-time quantitative fluorescent PCR(qRT-PCR).miR-NC mimic,miR-567 mimic,oe-NC,and oe-CDK8 were transfected into A549 and H1975 cells.The ex-pressions of miR-567 and CDK8 were detected using qRT-PCR.Cell proliferation was detected by CCK-8 method,and cell migration was detected by Transwell assay.Cell cycle changes were detected by flow cytome-try.The targeting of miR-567 and CDK8 was detected by luciferase reporter gene assay.Results In the tumor tissues of NSCLC patients,the expression of miR-567 was decreased,while the expression of CDK8 was in-creased,and the two were negatively correlated(P<0.05).In A549 and H1975 cells,miR-567 mimic group was compared with miR-NC mimic group,the expression of miR-567 was increased,the expression of CDK8 was decreased,the proliferation and migration levels of cells were decreased,the proportion of G1 phase was increased,and the proportion of S phase was decreased.The fluorescence intensity of miR-567 mimic group was lower than that of miR-NC mimic group in normal CDK8.miR-567 mimic+oe-CDK8 group was compared with miR-567 mimic+oe-NC group,the expression of CDK8 was increased,the proliferation and migration levels of cells were increased,the proportion of cells in G1 phase was decreased,and the proportion of cells in S phase was increased.Conclusion miR-567 can inhibit NSCLC proliferation and migration by targeting CDK8 expression and controlling tumor cell arrest in the S phase.
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Objective To screen long non-coding RNA(lncRNA)associated with disulfidptosis and investigate the immune landscape between lncRNA and pancreatic cancer,for effective guidance in clinical practice.Methods The normal and pancreatic cancer tissue samples were obtained from The Cancer Genome Atlas database,and the lncRNA associated with disulfidptosis was identified based on the Cox and LASSO regression analyses.A risk prognosis model was constructed,and its predictive performance was verified using comprehensive methods.An accurate nomogram was construted to predict the prognosis of patients with pancreatic cancer.The biological differences were analyzed via Gene Ontology,Gene Set Enrichment Analysis,and an immunoassay.The immunotherapy response was estimated using the tumor mutational burden(TMB)score.Results A total of 251 disulfidptosis-related lncRNAs were successfully identified,and three groups of lncRNAs were selected as the reference for the risk model.Pathway analysis showed that immune-related pathways were associated with disulfidptosis-related lncRNA risk models.The risk score was significantly correlated with immune cell infiltration and the ESTIMATE score.Patients with higher risk scores had elevated TMB,indicating that high-risk patients exhibited a better immune checkpoint blockade response.Conclusion The findings of this study contribute to a deeper understanding of disulfidpto-sis-related lncRNA and provide a potential therapeutic strategy for pancreatic cancer.
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Objective To investigate the expression of CD276 in pan-cancer,its effect on patient prognosis,and its mechanism of action.Methods TCGA and GTEx databases were used to study the differential expression of CD276 in tumor and normal tissues;R language was used to analyze the effects of CD276 on the prognosis of patients with pan-cancer,the tumor microenvironment,the tumor mutation burden,and microsatellite instability;the correlation between the expression of CD276 in tumor tissues and the tumor immune cell infil-tration was further examined;the biological function of CD276 was analyzed using Gene Set Enrichment Analysis(GSEA);and finally,real-time PCR was used to validate the expression of CD276 in tumors in vitro.Results A statistical difference in CD276 expression was detected between tumor and normal tissues(P<0.05).CD276 affects tumor prognosis and is strongly associated with the tumor microenvi-ronment,tumor mutational burden,microsatellite instability,and tumor immune cell infiltration.Conclusion Comprehensive pan-cancer analysis identified CD276 as a biomarker for immune infiltration and poor cancer prognosis,which can be used as a novel immunomodula-tory target for the development of immunotherapeutically-targeted drugs,providing new ideas for tumor therapy.
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@#[Objective[ To analyze the main syndrome types, medication rules, and core prescription characteristics of traditional Chinese medicine (TCM) in the treatment of metabolism-associated fatty liver disease (MAFLD), and to predict the anti-MAFLD mechanism of core formula, so as to provide references for the clinical application of TCM and the development of new drugs. [Methods] Literature research on TCM in treating MAFLD was retrieved from China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), and Wanfang Database since the establishment of the database to July 2022. Excel 2019 and Chinese Medicine Inheritance Computing Platform (V3.0) were used for frequency analysis, association rule analysis, and cluster analysis of effective prescriptions. The key components, targets, and action pathways of anti-MAFLD core formulas were predicted by network pharmacology. Finally, the interactions between the obtained core components and their core targets were verified reversely by molecular docking technology. [Results] A total of 218 articles were screened and selected, including 352 prescriptions, involving 270 traditional Chinese herbs. The drugs were used a total of 3 901 times, and a total of 10 915 cases were collected, among which the prevalence rate was higher in males. The main types of TCM syndrome included intermingled phlegm and blood stasis syndrome, liver depression and spleen deficiency syndrome, and damp-heat in liver and gallbladder syndrome, among which Shanzha (Crataegi Fructus), Danshen (Salviae Miltiorrhizae Radix et Rhizoma), Fuling (Poria), Zexie (Alismatis Rhizoma), Chaihu (Bupleuri Radix), and Baizhu (Atractylodis Macrocephalae Rhizoma) were the most frequently used. The properties of Chinese medicine primarily encompassed thermal characteristics, with a predominant emphasis on cold and warm; the flavors of herbs were predominantly characterized by bitterness and sweetness, while the majority exhibited tropism towards the spleen and liver meridians. The drugs were primarily classified based on their efficacy in tonifying deficiencies, promoting diuresis and moistening, enhancing blood circulation and removing blood stasisheat-clearing, etc. The association rules were employed to derive a set of 20 core drug pairs, while cluster analysis was utilized to identify three distinct groups of core drug combinations. Network pharmacological showed that the main components of the core formula “Shanzha (Crataegi Fructus) - Danshen (Salviae Miltiorrhizae Radix et Rhizoma) - Zexie (Alismatis Rhizoma) - Chaihu (Bupleuri Radix) - Fuling (Poria)” in the treatment of MAFLD were quercetin, apigenin, puerarin, luteolin, ursolic acid, kaempferol, tanshinone IIA, emodin, paeonol, etc., which involved RAC-alpha serine/threonine-protein kinase 1 (AKT1), cellular tumor antigen p53 (TP53), interleukin (IL)-6, IL-1β, signal transducer and activator of transcription 3 (STAT3), epidermal growth factor receptor (EGFR), peroxisome proliferative activated receptor gamma (PPARG), and other key targets. The molecular docking results showed that the core components had good binding to lipid and atherosclerosis, and phosphatidylinositol 3 kinase (PI3K)/AKT signaling pathway-associated proteins. [Conclusion] The main principles of TCM for the treatment of MAFLD involve soothing the liver and strengthening the spleen, eliminating phlegm and dampness, clearing heat and dampness, as well as promoting blood circulation and removing blood stasis. The core formula may exert anti-MAFLD effects mediated through multiple components, targets, and signaling pathways. This study establishes a theoretical foundation for the clinical application of TCM in the treatment of MAFLD, and serves as a reference for further exploration of new drugs against MAFLD.
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ObjectiveTo explore whether Hei Xiaoyaosan can inhibit the inflammatory response in the hippocampi of Alzheimer's disease (AD) rats by regulating and activating the Wnt/β-catenin signaling pathway to improve the cognitive and memory dysfunction. MethodAmong the 90 male Wistar rats, 12 were randomly selected as the blank group (normal saline) and 12 as the sham operation group (normal saline). For the remainder, amyloid β-protein42 (Aβ42) was injected in the left and right hippocampus to induce AD, and then the AD rats were randomized into model group, low-, medium-, and high-dose Hei Xiaoyaosan groups (corresponding doses of Hei Xiaoyaosan, ig), and donepezil group (donepezil hydrochloride,ig), with 12 in each group. The administration lasted 42 days. The pathological changes of hippocampal CA1 region was observed based on Nissl staining. The escape latency on the 1st to 5th day in Morris water maze was recorded and the spatial memory on the 6th day was tested. Enzyme-linked immunosorbent assay (ELISA) was employed to detect the expression of interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α) in rat hippocampus and serum, Western blotting to examine the protein expression of glycogen synthase kinase-3β (GSK-3β), β-catenin, and peroxisome proliferator-activated receptor gamma (PPARγ), and real-time polymerase chain reaction (Real-time PCR) to determine the mRNA expression of rat GSK-3β, β-catenin, and PPARγ. ResultCompared with the blank group, the number of neurons in the hippocampal CA1 area of model group was significantly reduced, the arrangement was uneven, the cell body was damaged more obviously, and the Neisser body was unclear. The treatment group was significantly prolonged (P<0.01), and the number of crossing stations was significantly reduced (P<0.01), the levels of IL-10 in serum and hippocampus of rats in the model group were significantly decreased, while the levels of IL-6 and TNF-α were significantly increased (P<0.01), the GSK-3β protein and mRNA in the model group were significantly increased, and the protein expressions of β-catenin and PPARγ were significantly decreased (P<0.01), and the difference was more obvious. The number of neurons in the donepezil group was more distributed, neatly arranged, the structure was intact, and the Nissl bodies were clear and definite, the escape latency on the 3rd to 5th days in middle and high dose groups of Hei Xiaoyaosan and the donepezil group was significantly shortened (P<0.01), the number of crossing platforms increased significantly (P<0.01), the expression levels of IL-10 in the rat hippocampus and serum were significantly increased, while IL-6 and TNF-α were significantly decreased (P<0.01), GSK-3β in the rat hippocampus was significantly increased. The expressions of GSK-3β protein and mRNA were significantly decreased, while the expression levels of β-catenin and PPARγ protein and mRNA were significantly increased (P<0.01). There was no significant difference in each index between the donepezil hydrochloride group and the high-dose Hei Xiaoyaosan group. ConclusionHei Xiaoyaosan can inhibit the inflammatory response in the hippocampus of AD rats by regulating the Wnt/β-catenin signaling pathway, thereby alleviating the cognitive and memory impairment of AD rats.
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OBJECTIVE@#To investigate the effect and safety of Guanxinning Tablet (, GXN) for the treatment of stable angina pectoris patients with Xin (Heart)-blood stagnation syndrome (XBSS).@*METHODS@#One hundred and sixty stable angina pectoris patients with XBSS were randomly assigned to receive GXN (80 cases) or placebo (80 cases, Guanxinning simulation tablets, mainly composed of lactose), 4 tablets (0.38 g/tablet), thrice daily for 12 weeks. After treatment, an exercise stress test (treadmill protocol), Chinese medicine (CM) syndrome score, electrocardiogram (ECG), and nitroglycerin withdrawal rate were evaluated and compared in the patients between the two groups. Meanwhile, adverse events (AEs) were evaluated during the whole clinical trial.@*RESULTS@#Compared with the control group, the time extension of exercise duration in the GXN group increased 29.28 ±17.67 s after treatment (P>0.05); moreover, the change of exercise duration in the GXN group increased 63.10 ±96.96 s in subgroup analysis (P<0.05). The effective rates of angina pectoris, CM syndrome and ECG as well as nitroglycerin withdrawal rate were 81.33%, 90.67%, 45.76%, and 70.73%, respectively in the GXN group, which were all significantly higher than those in the control group (40.58%, 75.36%, 26.92%, 28.21%, respectively, P<0.05).@*CONCLUSION@#GXN was a safe and effective treatment for stable angina pectoris patients with XBSS at a dose of 4 tablets, thrice daily.
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OBJECTIVE Shenfu injection(SFI) is an effective treatment of cardiogenic shock, the pathology of the central link was microcirculation disturbance. However, whether the microcirculation status of the early- and mid-stage of cardiogenic shock has any difference is unclear. This study aimed to observe the effect of SFI on the microcirculatory disturbance in mesentery for early- and mid-stage of cardiogenic shock rat. METHODS The early- and mid-stage model of cardiogenic shock was estab?lished by ligating the ending or root of left anterior descending coronary arteries(LADCA). The rats were randomly divided into 9 groups, ie control group, early- stage model group, mid- stage model group, 3 early medicated groups and 3 mid medicated groups(the dosage was 1, 3.33, 10 mL·kg-1 SFI for cardiogenic shock rats of early- and mid-stage, respectively). Parameters in mesenteric microcircu?lation, such as velocity of RBCs in venules, diameters of venules, the count of leukocyte adhesion and vascular permeability which calculated by FITC-dextran leakage were observed through an Geneandi-M2 inverted intravital microscope and high-speed video camera system. RESULTS The cardiogenic shock induced by ligating the LADCA resulted in a number of responses in microcirculation, including a significant increase in the counts of adhesive leukocytes, narrowing of the vascular diameter, decrease in the velocity of RBCs and dextran efflux. All of the above parameters for early- stage cardiogenic shock rats were attenuated by the treatment with SFI, especially the dosage of 10 mL·kg-1. While SFI had no apparent time- effect on the vascular diameter and vascular permeability in mesentery for mid-stage cardiogenic shock rats. CONCLUSION The microcirculation status of the early- and mid-stage of cardiogenic shock rats were quite different. The efficacy of early treatment with SFI was more obvious than the mid administration, which could provide experimental and theoretical basis for the patients with cardiogenic shock in an earlier time.
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To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
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Animals , Female , Humans , Pregnancy , Animals, Genetically Modified , Genetics , Fibroblasts , Gene Knock-In Techniques , Gene Knockout Techniques , Goats , Genetics , Lactoferrin , Genetics , Lactoglobulins , Genetics , Milk , Chemistry , Nuclear Transfer Techniques , Plasmids , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To observe the clinical efficacy of treating patients with coronary slow flow phenomenon by Yiqi Huoxue Recipe (YHR) combined Western drugs, thus providing clinical evidence for further studies.</p><p><b>METHODS</b>Totally 61 patients with coronary slow flow phenomenon were randomly assigned to the treatment group (31 cases) and the control group (30 cases). Patients in the control group were treated with basic treatment of Western medicine, while those in the treatment group were treated with basic treatment of Western medicine and YHR. The therapeutic course for all was two months. Clinical symptoms were observed, and electrocardiogram examinations taken, and left ventricular ejection fraction (LVEF) were evaluated before treatment and at two months after treatment.</p><p><b>RESULTS</b>Patients' clinical symptoms and electrocardiogram examinations were significantly improved in the treatment group. Its effective rate of improved symptoms was 90.32% in the treatment group, superior to that in the control group (76.67%, P < 0.05). The effective rate of electrocardiogram examinations was 87.10% in the treatment group, superior to that in the control group (73.33%, P < 0.05). But there was no statistical difference in LVEF between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>YHR combined Western drugs could improve clinical symptoms and electrocardiographic ischemia in patients with coronary slow flow phenomenon.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Blood Flow Velocity , Coronary Artery Disease , Drug Therapy , Drug Therapy, Combination , Drugs, Chinese Herbal , Therapeutic Uses , Myocardial Ischemia , Drug Therapy , Phytotherapy , Treatment OutcomeABSTRACT
Objective To investigate the clinical features of aortic dissection (AD) and emergency treatments. Methods Data from 784 patients with aortic dissection were collected in the Department of Emergency from January 2000 through December 2009. A retrospective analysis was carried out to determine the survival rate, mortality rate and treatment efficiency. Results Pain was the most common onset symptom (77.7% , 609/784). The majority of patients (86.5%) had essential hypertension (678/784). All the patients with preoperative diagnosis of aortic dissection underwent emergency medical intervention by internists resulting in 81.5% survival rate (639/784) and 18.5% mortality rate (145/784). There were 157 patients without improvement (20.0% ) and the total efficiency rate was (83. 1% ). The efficiency rate of conventional treatment was 76.4% , while the efficiency rate of triple four-procedure treatment was 89. 8% (P<0.05). Of them, 139 patients (17. 7% ) died in the hospital. Among them,. 26 patients died within 24 hours (18.4% ) and 47 cases died within 48 hours (33. 8% ) and 66 patients died within 72 hours (47.2% ). There were 92 patients who refused treatments after diagnosis, and among them, 81 patients died within 72 hours (88.04% ). The difference in mortality rate between two groups was significant (P<0.05). Conclusions The diagnosis of aortic dissection depends on detailed history, physical examination and CT or MRI imaging. Analgesia, sedation and control of blood pressure are essential for emergency treatments. Early diagnosis and effective emergency treatments are the critical strategy for the early surgical intervention and time window for further treatment to improve the survival rate of AD.
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OBJECTIVE@#To construct a subtracted cDNA library of chronic intermittent hypoxia (CIH) rabbit liver by suppression subtractive hybridization (SSH).@*METHODS@#Twenty-four rabbits were divided into 4 groups: ordinary feeding group, full-fat food group, ordinary feeding in chronic intermittent hypoxia group, and full-fat food in chronic intermittent hypoxia group. The mRNAs were extracted from different rabbit livers and converted into double-strand cDNA. After digestion with restriction enzyme, the cDNA of hyperlipidemia-sensitive rabbit group was subdivided into 2 portions and each one was lighted with different adaptors. Two rounds of both hybridization and suppression PCR obtained the differentially expressed cDNA. The PCR products were inserted into T/A vector to set up the subtractive cDNA library. The clones were selected and amplified by PCR and identified.@*RESULTS@#Based on the pathology of the abdominal aorta and liver, and the amplified library contained 500 positive bacteria clones, including 462 clones, which had inserts from 250 to 700 bp by PCR analysis. A novel rabbit gene, Cthrc1, involved in CHI had been cloned. The GenBank Accession Number is XM_418373.@*CONCLUSION@#The molecular mechanism of CIH promoting atherogenesis formation is made clear.
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Animals , Rabbits , Gene Library , Genetic Vectors , Hyperlipidemias , Hypoxia , Metabolism , Pathology , Liver , Metabolism , Pathology , Nucleic Acid Hybridization , Methods , RNA, Messenger , GeneticsABSTRACT
@#ObjectiveTo observe the effect of Tongxinfang on coronary collateral formation and stent-restenosis in patients with coronary artery disease after stenting.Methods61 patients with coronary artery disease were randomly divided into the treatment group (n=30), and control group (n=31). Patients of the treatment group were taking Chinese medicine Tongxinfang for six months, but cases of the control group only received basic treatment. Coronary collateral formation, stent-restenosis and ejection fraction (EF) of patients of two groups were assessed through aniograms and echocardiography before treatment and after six months.ResultsCoronary collateral formation and EF of patients of the treatment group improved significantly compared with cases of the control group (P<0.01 or P<0.05), but the minimal lumen diameter and percent of diamter stenosis were not significantly different between two groups (P>0.05).ConclusionTongxinfang can improve coronary collateral formation and EF.
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@#ObjectiveTo observe the effect of Yiqihuoxue recipe on left ventricular remodeling after acute myocardial infarction (AMI).Methods40 patients with AMI were randomly divided into treatment group and control group with 20 cases in each group and received Yiqihuoxue recipe or fasinopril respectively for 6 months. Changes of clinical symptoms, the level of plasma angiotensin Ⅱ(ATⅡ), aldosterone (ALD), endothelin (ET) and echocardiographical indexes: left ventricular end-diastolic volume index (LVEDVI), left ventricular end-systolic volume index (LVESVI), ejection fraction (EF), left ventricular mass index (LvmassI), wall motion index (WMI), wall thick (WT), E/A were assessed before discharge, and in the end of 3rd and 6th month after AMI.ResultsClinical symptoms of patients of treatment group improved significantly (P<0.01). EF and E/A of all patients in two groups increased (P<0.01 or P<0.05), LvmassI and WT reduced (P<0.05), but there were no significantly differences between two groups in LVEDVI, LVESVI, EF, LvmassI, WMI, E/A and WT(P>0.05). The level of plasma ET decreased in treatment group (P<0.05), the level of plasma ATⅡ and ALD of control group decreased more than that of treatment group (P<0.05).ConclusionYiqihuoxue recipe can significantly improve clinical symptoms, cardic function, and left ventricular remodeling, showing a better clinical efficacy.