1.Serum miR-203 expression level detected by fluorescence quantitative PCR in cervical cancer patients and its significance
Chunlan YANG ; Xianjuan SHEN ; Shaoqin JU ; Jianyou SU
International Journal of Laboratory Medicine 2015;(24):3552-3553
Objective To establish the method of the SYBR Green Ⅰ real‐time fluorescent quantitative PCR for detecting the se‐rum miR‐203 expression level ,and to detect the serum miR‐203 expression levels in the patients with cervical cancer ,cervical benign diseases and healthy controls .Methods The miR‐203 ,U6 stem loop RT primers and the PCR amplification primers were designed for conducting fluorescence quantitative PCR ,with U6 as the internal relative quantification ,the serum miR‐203 levels were com‐pared among different cervical diseases .Results The established method could specifically detect the amplification signal of serum miR‐203 ,the melting curve was single and PCR products were specific .The serum miR‐203 level in the patients with cervical cancer was significantly higher than that in the patients with benign cervical diseases such as hysteromyoma and cervicitis ,the difference was statistically significant (P<0 .05) .Conclusion The SYBR Green Ⅰ real‐time fluorescent quantitative PCR is a quick ,simple detection method with high sensitivity and good specificity ,which may have a better application prospect in cervical cancer auxiliary diagnosis .
2.Anti-inflammation and Regulation of HIF-1α/VEGF of 3 Kinds of Borneolum in Prevention and Treatment of AMI Model Rats
Liying WANG ; Jian WANG ; Yamei FAN ; Jiajun WANG ; Yin FU ; Xianjuan YANG ; Daoyin GONG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(19):61-72
ObjectiveTo investigate the possible mechanism of different doses of L-Borneolum,Borneolum,and Borneolum Syntheticum in the electrophysiology,anti-inflammation,and regulation of hypoxia inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF) cardiovascular protection of the experimental acute myocardial infarction (AMI) rats. MethodSD male adult rats were randomly divided into thirteen groups according to their body weight,namely the sham operation group,the model group,the solvent model group,the nitroglycerin group,the Borneolum high,medium,and low-dose (0.6,0.3, 0.15 g·kg-1) groups,the L-Borneolum high,medium,and low-dose (0.2,0.1, 0.05 g·kg-1) groups,and the Borneolum Syntheticum high,medium,and low-dose (0.2,0.1, 0.05 g·kg-1) groups,with 10 rats in each group. Rats were given 10 mL·kg-1 by gavage for 3 d of pre-administration. Thirty minutes after the last administration,the left anterior descending coronary artery (LAD) was ligated to induce the model,and the successful rat model was continuously treated for 3 d. BL-420N biosystem was used to analyze the electrocardiogram (ECG) and heart rate variability (HRV) before and after modeling and after 3 d of treatment. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to determine the mRNA expressions of interleukin-1β (IL-1β) and interleukin-6 (IL-6) in the myocardial tissue Western blot and immunohistochemistry were used to determine the protein expression levels of VEGF receptor 1 (VEGFR1),HIF-1α,and CD34. ResultCompared with the sham operation group,the model group significantly increased the heart rate,ECG ST wave,T wave,QRS duration,QTC interval,and Q wave on the day of modeling and after 3 d of treatment,and significantly changed HRV and T wave (P<0.05,P<0.01). As compared with the solvent model group,on the day of modeling,the heart rate of the L-Borneolum medium and low-dose groups and the Borneolum groups,the ST wave of the L-Borneolum groups,the Borneolum high and medium-dose groups,and the Borneolum Syntheticum high-dose group,HRV parameters of the L-Borneolum groups,the Borneolum medium and low-dose groups,and the Borneolum Syntheticum high-dose group,LF/HF of the L-Borneolum high and medium-dose group,the Borneolum low-dose group,and the Borneolum Syntheticum groups,T wave of the L-Borneolum high-dose group,the Borneolum Syntheticum high-dose group,and Borneolum medium-dose group,QTC interval of the L-Borneolum medium and low-dose groups and the Borneolum high and medium-dose groups,and QRS duration of the L-Borneolum high and low-dose groups,the Borneolum high and low-dose groups,and the Borneolum Syntheticum groups were significantly reduced or shortened (P<0.05,P<0.01). After 3 d of treatment,the heart rate of the L-Borneolum groups,the Borneolum high and medium-dose groups,and the Borneolum Syntheticum medium-dose group,ST wave of the L-Borneolum group,the Borneolum high and medium-dose groups,and the Borneolum Syntheticum high-dose group,OTC interval,ORS duration,and Q wave of the L-Borneolum high-dose group,the Borneolum high-dose group,and the Borneolum Syntheticum high and medium-dose groups,QRS duration of the L-Borneolum medium-dose group,QTC interval of the Borneolum medium-dose group,and Q wave of the Borneolum Syntheticum low-dose group were all significantly reduced or shortened(P<0.01). The mRNA expressions of IL-1β and IL-6 in the L-Borneolum medium and low-dose groups,the Borneolum medium and low-dose groups,and the Borneolum Syntheticum high and medium-dose groups were significantly down-regulated(P<0.01),and LF/HF in the L-Borneolum high and medium-dose groups,the Borneolum high and medium-dose groups,and the Borneolum Syntheticum high and low-dose groups were significantly reduced (P<0.05,P<0.01). HRV in the L-Borneolum high-dose group,the Borneolum groups,and the Borneolum Syntheticum high and low-dose groups,and T wave in the Borneolum high and medium-dose groups and the Borneolum Syntheticum high-dose group were increased significantly. The protein expressions of HIF-1α,VEGFR1,and CD34 in the L-Borneolum medium and low-dose groups,the Borneolum low-dose group,and the Borneolum Syntheticum high-dose group were significantly up-regulated,as well as those of VEGFR1 and CD34 in the Borneolum medium-dose group (P<0.05,P<0.01). ConclusionThe 3 kinds of Borneolum improves the heart rate,heart rate variability,and electrocardiogram of AMI model rats to different degrees,and may play a myocardial protective effect by anti-inflammation and promotion of angiogenesis. The combined effect suggests that L-Borneolum has the superior effect next to Borneolum,and Borneolum Syntheticum has the inferior effect.
3.Mechanism of Huanglian Houpotang on Ulcerative Colitis by Network Pharmacology and Experimental Verification
Jiajun WANG ; Xianjuan YANG ; Liying WANG ; Yin FU ; Jian WANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):217-224
ObjectiveTo explore the active components and underlying mechanism of Huanglian Houpotang (HHD) against ulcerative colitis(UC) based on network pharmacology and animal experiments. MethodThe active components of HHD were preliminarily obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) and screened out by TCMSP, SwissADME, and SwissTargetPrediction, and their targets were predicted. Relevant microarrays were searched for disease genes with the help of Gene Expression Omnibus (GEO). The common targets of HHD and disease genes were screened out to obtain the potential targets of HHD against UC. The drug-active component-target-disease network was constructed using Cytoscape 3.7.2. The potential therapeutic targets were imported into the DAVID 6.8 for GO-Biological process (GO-BP) analysis to predict related biological processes which were subsequently verified by the animal experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the effect of HHD on inflammatory factors in colon tissues of mice. Western blot was used to detect the protein expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteinyl aspartate-specific protease 3 (Caspase-3). The IVIS system was used to detect the content of reactive oxygen species (ROS) in colon tissues of mice in each group. ResultNineteen active components of HHD were screened out, involving 32 potential therapeutic targets against UC and 158 biological processes. The results of the animal experiment showed that HHD exerted its anti-UC effect by inhibiting the expression of inflammatory factors tumor necrosis factor (TNF)-α and interleukin-1β (IL-1β), reducing the content of apoptotic proteins, and regulating the expression of ROS. ConclusionThis study revealed the rationality of predictions and guidance of network pharmacology in experimental design, and confirmed that HHD could exert its effects by participating in biological processes such as immune inflammation, apoptosis, and ROS, which is expected to provide a basis for the mechanism research of HHD in the treatment of UC.
4.Analysis of gene expression difference in acute myocardial infarction based on bioinformatics and prediction of traditional Chinese medicine
Xianjuan Yang ; Liying Wang ; Jian Wang ; Jiajun Wang ; Yin Fu ; Jinxiu Li ; Linxuan Xiao
Acta Universitatis Medicinalis Anhui 2022;57(1):15-20
Objective :
To analyze and compare the gene chip data of normal people and patients with acute myocardial infarction through GEO gene expression database , to screen out differentially expressed genes (DEGs) , and to predict potential Chinese medicines for the treatment of acute myocardial infarction.
Methods :
GSE66360 gene microarray was downloaded , DEGs information was obtained by analysis , gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differential genes were performed , key genes were further analyzed by String database and Cytoscape software , and key genes were mapped to the medical ontology information retrieval platform (Coremine Medical) to screen potential Chinese medicines for the treatment of acute myocardial infarction.
Results :
A total of 943 differentially expressed genes were screened. The biological process was mainly
enriched in myeloid leukocyte activation , regulation of cytokine production , leukocyte chemotaxis , etc. The cellular component was mainly focused on secretory granule lumen , membrane surface , and extrinsic components of the membrane , etc. Molecular function was mainly in chemokine receptor binding , pattern recognition receptor activity , cytokine binding , etc. KEGG analysis showed that the main signaling pathways involved were tumor necrosis factor (TNF) , hypoxia inducible factor⁃1 (HIF⁃1) , and JAK⁃STAT signaling pathways , etc. The key genes to be screened are formyl peptide receptor 2 (FPR2) , signal transducer and activator of transcription 3 (STAT3) , chemokine (C - X - C motif) ligand 1 ( CXCL1) , chemokine ( C ⁃X ⁃C motif) ligand 8 ( CXCL8) , ubiquitin protein ligase E3 component n⁃recognin 4 ( UBR4 ) , jun proto⁃oncogene ( JUN ) , platelet⁃activating factor receptor (PTAFR) , Fc fragment of IgE , high affinity I , receptor for; gamma polypeptide (FCER1G) , G protein⁃coupled
receptor 84 (GPR84) , plasminogen activator, urokinase (PLAU) . The potential herbs predicted for the treatment of acute myocardial infarction were Centipede (P = 0. 003 30) , Rithoma Curcuma (P = 0. 002 39) , Curcuma (P = 0. 002 40) , Paris polyphylla Smith (P = 0. 002 48) , Salviae miltiorrhizae (P = 0. 002 72) , Fritillary bulb (P = 0. 003 71) , and Panax ginseng (P = 0. 001 59) .
Conclusion
Traditional Chinese medicines such as Rithoma curcuma in activating blood and removing blood stasis medicine , Panax ginseng in nourishing Qi , and Fritillary bulb in medicine for the treatment of cough and asthma have protective effects on acute myocardial infarction. The mechanism of action may be related to the regulation of immune and anti⁃inflammatory signaling pathways.