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Understanding the research methods for drug protein targets is crucial for the development of new drugs, clinical applications of drugs, drug mechanisms, and the pathogenesis of diseases. Cellular thermal shift assay (CETSA), a target research method without modification, has been widely used since its development. Now, there are various CETSA-based technology combinations, such as mass spectrometry-based cellular thermal shift assay (MS-CETSA), isothermal dose response-cellular thermal shift assay (ITDR-CETSA), amplified luminescent proximity homogeneous assay-cellular thermal shift assay (Alpha-CETSA), etc., which combine their respective advantages and further expand the application scope of CETSA. These technologies are suitable for the entire drug development chain, from drug screening to monitoring the target binding and off-target toxicity of drugs in patients. Based on the author's research experience, this paper reviews the principles of CETSA and related binding technologies, their application in target discovery, and the progress of data processing and analysis in recent years, aiming to provide reference and reference for the further application of CETSA.
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Parkinson's disease is a common degenerative disease of central nervous system. Exercise for Parkinson's disease includes Taiji Quan, treadmill, dance and balance training, etc. Taiji Quan exercise is mainly for balance function, and treadmill training for gait obstacles. Dance can improve not only balance and gait, but also cognition and emotion. The possible molecular mechanism may involve to adjust the content of dopaminergic neurons in the nigrostatin, increase the content of non-dopaminergic transmitters, increase brain-derived neurotrophic factors, promote the plasticity of nerve cells, reduce oxidative stress, enhance mitochondrial activity, and activate the immune system, etc., to slow the degeneration of dopaminergic neurons and improve the symptoms of Parkinson's disease.
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Objective:Screen out the antitumor constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma base on system pharmacology with chemical constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma as study objects, in order to provide the theoretical basis for the development of antitumor and nontoxic activities of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma. Method:The small molecule ligand library of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma was built based on Traditional Chinese Medicine Systems Pharmacology(TCMSP), energy of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma was matched with the key protein targets of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signal pathway by molecular docking (SYBYL2.1, Tripos), the Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma-targets network model was established based on Cytoscape 3.5.1, and the physicochemical properties of the antitumor activity in Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma were predicted by using SwissADME and admetSAR. Result:There were 25 small molecule constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma. Through the energy match, key antitumor constituents of Pinelliae Rhizoma were gondoic acid, 10,13-eicosadienoic, baicalin, 12,13-epoxy-9-hydroxynonadeca-7,10-dienoic acid. Key antitumor constituents of Aconiti Lateralis Radix Praeparata were deltoin, sitosterol, neokadsuranic acid B, 11,14-eicosadienoic acid. Phosphatidylinositol 3-kinase (PI3Kα), phosphatase and tensin homolog deleted on chromosome ten (PTEN), phosphoinositide dependent protein kinase 1 (PDK1) were key antitumor targets of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma. There were 8 key antitumor constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma, which had a low CYP450 inhibition and basically followed the Lipinski rule. Conclusion:Antitumor nontoxic constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma and key targets are screened out from the molecular level, which provides the new ideas for the effective use of nontoxic traditional Chinese medicine(TCM) and breaks the restrictions in using nontoxic TCM.
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Objective:To evaluate the reliability and validity of Chinese version of International Parkinson and Movement Disorder Society (MDS)-Unified Parkinson's Disease Rating Scale (UPDRS), and then do comparative analysis with original UPDRS. Methods:From May, 2016 to December, 2018, 225 patients with Parkinson's disease were respectively evaluated by MDS-UPDRS and UPDRS, and then the reliability and validity of MDS-UPDRS were evaluated. Correlation between two scales was analyzed. Results:The Cronbach's coefficient of the total MDS-UPDRS was 0.734. Rank correlation's coefficient was 0.905. For factor analysis, KMO was 0.728. Pearson's correlation analysis showed that the correlation's coefficient of the common factor of the two scales was 0.969, while the correlation's coefficient of the total score of the two scales was 0.972. Compared with UPDRS, MDS-UPDRS added a number of non-motor symptoms, which adjusted the standard of measurement and increased the accuracy degree of evaluation at early stage of disease. Besides, MDS-UPDRS added a lot of instructions to evaluate the standards to make the evaluation normatively. Conclusion:Chinese version of MDS-UPDRS has high reliability and validity, and highly correlate with the original UPDRS. It is more suitable for Chinese patients with Parkinson's disease in clinical evaluation than UPDRS.
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<p><b>OBJECTIVE</b>To explore the correlation between the recurrence of cerebral infarction and aspirin resistance (AR)/Chinese medical (CM) constitutions.</p><p><b>METHODS</b>Totally 413 cerebral infarction patients took Aspirin Enteric-coated Tablet (100 mg per day) while receiving routine therapy, 5 days at least in a week. They were followed-up for 12 months. Aspirin sensitivity (AS) was determined using turbidimetry. CM constitutions among patients with different AS were compared. Ratios of AR patients and AS patients of different CM constitutions in cerebral infarction recurrent patients were compared. Platelet membrane glycoproteins (GP) II b HPA-3 gene polymorphism was detected by polymerase chain reaction (PCR) method. Correlation between recurrence of cerebral infarction and AR, bb genotypes, CM constitutions times AS were analyzed by Logistic regression.</p><p><b>RESULTS</b>Totally 11 patients dropped out, 101 (25.12%)with recurrent cerebral infarction and 301 (74.88%) without recurrent cerebral infarction. There were 152 (37.81%) AR patients and 250 (62.19%) AS patients. AR accounted for 26.6% (80/ 301) and AS accounted for 73.4% (221/301) in non-recurrent cerebral infarction patients. AR accounted for 71.3% (72/101) and AS accounted for 28.7% (29/101) in recurrent cerebral infarction patients. There was statistical difference in AR and AS ratios (χ2 = 64.287, P = 0.000). The proportion of yin deficiency constitution (YDC) was the largest [28.3% (43/152)] in AR patients. The proportion of blood stasis constitution (BSC) was the largest [23.6% (59/250)] in AS patients. There was statistical difference in CM constitutions between AR patients and AS patients (χ2 = 21.574, P < 0.01). The former 4 recurrent rates occurred in AR patients of YDC, BSC, damp-phlegm constitution (DPC), qi deficiency constitution (QDC). YDC occupied the first place [22.4% (34/152)]. The former 4 recurrent rates occurred in AS patients of BSC, QDC, DPC, damp-heat constitution (DHC). BSC occupied the first place [3.2% (2/250)]. Compared with non-recurrent cerebral infarction patients and AS patients, bb gene occurred most often, but aa gene and ab gene occurred obviously lesser in non-recurrent cerebral infarction patients and AR patients (χ2 = 20.171, χ2 = 55.139, P < 0.01). AR and bb gene were positively correlated with recurrent cerebral infarction (OR = 18.423, P = 0.000; OR = 1.304, P = 0.028). Body constitutions interacted with AS (OR = 0.707, P = 0.000).</p><p><b>CONCLUSIONS</b>Recurrent cerebral infarction was closely related to AR and constitutional types. The recurrence rate was higher in AR patients of YDC. GP I b HPA-3 bb genotype might be a risk factor for AR and recurrent cerebral infarction.</p>
Subject(s)
Humans , Aspirin , Therapeutic Uses , Body Constitution , Cerebral Infarction , Drug Resistance , Medicine, Chinese Traditional , Neoplasms , Recurrence , Yin DeficiencyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of short hairpin RNA (shRNA) targeting N-acetylglucosaminyltransferase V (GnT-V) on the proliferation of prostate cancer PC-3 cells.</p><p><b>METHODS</b>Lipofectamine 2000 was used to transfect the recombinant plasmids carrying the shRNA targeting GnT-V gene into PC-3 cells. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to measure the mRNA expression of GnT-V, and CCK-8 assay was used to measure the cell proliferation after the transfection.</p><p><b>RESULTS</b>The recombinant plasmids were successfully transfected into PC-3 cells, resulting in a reduction of GnT-V mRNA expression by 73%. The proliferation of PC-3 cells was significantly inhibited after the transfection.</p><p><b>CONCLUSION</b>The shRNA targeting GnT-V gene can reduce the expression of GnT-V mRNA and inhibit the proliferation of PC-3 cells in vitro.</p>
Subject(s)
Humans , Male , Cell Line, Tumor , Cell Proliferation , N-Acetylglucosaminyltransferases , Genetics , Prostatic Neoplasms , Genetics , Pathology , RNA Interference , RNA, Small Interfering , Genetics , TransfectionABSTRACT
To study the effect of soybean isoflavone on liver oxidative stress resulting from 60Co-gamma rays. Methods: Totally 80 normal female Kunming mice were evenly randomized into 5 groups according to body weight: 3 intervention groups, single irradiation group and normal control group. The normal group and single irradiation groups were given 0.5% CMC-Na, and the 3 intervention groups were given different doses of soybean isoflavone (50 mg/kg, 100 mg/kg, 400 mg/kg) respectively for 14 d. The whole body of single irradiation group and intervention groups were subjected to 4.56 Gy 60Co-γ radiation once on the 7th day, and then the mice were killed on the 2nd day and the 7th day after radiation. Results: The CAT activity of liver tissue of 100,400 mg/kg intervention groups and 3 SI groups were significantly increased on the 2nd day and 7th day after irradiation(P<0.05), respectively; the GSH-Px activity of 100 mg/kg SI group was significantly increased(P<0.05) on the 7th day after irradiation; the T-SOD activity of 50 mg/kg SI group was significantly decreased (P<0.05) on the 2nd day after irradiation,while no difference was observed among remaining groups. The MDA content of 100 mg/kg group was significantly decreased on the 7th day after radiation compared with control group, and MDA content of each group subjected to irradiation were increased on the 2nd day after irradiation,but 3 SI groups nearly decreased to normal level on the 7th day after irradiation. Conclusion: The soybean isoflavone can enhance the antioxidant capability of mice, but it does not show a dose-effect relationship.
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To study the effect of soybean isoflavone on liver oxidative stress resulting from 60Co-gamma rays. Methods: Totally 80 normal female Kunming mice were evenly randomized into 5 groups according to body weight: 3 intervention groups, single irradiation group and normal control group. The normal group and single irradiation groups were given 0.5% CMC-Na, and the 3 intervention groups were given different doses of soybean isoflavone (50 mg/kg, 100 mg/kg, 400 mg/kg) respectively for 14 d. The whole body of single irradiation group and intervention groups were subjected to 4.56 Gy 60Co-γ radiation once on the 7th day, and then the mice were killed on the 2nd day and the 7th day after radiation. Results: The CAT activity of liver tissue of 100,400 mg/kg intervention groups and 3 SI groups were significantly increased on the 2nd day and 7th day after irradiation(P<0.05), respectively; the GSH-Px activity of 100 mg/kg SI group was significantly increased(P<0.05) on the 7th day after irradiation; the T-SOD activity of 50 mg/kg SI group was significantly decreased (P<0.05) on the 2nd day after irradiation,while no difference was observed among remaining groups. The MDA content of 100 mg/kg group was significantly decreased on the 7th day after radiation compared with control group, and MDA content of each group subjected to irradiation were increased on the 2nd day after irradiation,but 3 SI groups nearly decreased to normal level on the 7th day after irradiation. Conclusion: The soybean isoflavone can enhance the antioxidant capability of mice, but it does not show a dose-effect relationship.
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<p><b>AIM</b>To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender.</p><p><b>METHODS</b>Semen samples were collected by electro-ejaculation from four sexually mature male cynomolgus monkeys. The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris mTTE synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1 %, 3 %, 5 %, 10 % and 15 % [v/v]) and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity.</p><p><b>RESULTS</b>The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P0.05) for 5 % glycerol (42.95 +/- 2.55 and 50.39+/- 2.42, respectively) than those of the other groups (1%: 19.19 +/- 3.22 and 24.84 +/- 3.64; 3%: 34.23 +/- 3.43 and 41.37 +/- 3.42; 10%:15.68 +/- 2.36 and 21.39 +/- 3.14; 15%: 7.47 +/- 1.44 and 12.90 +/- 2.18). The parameters for 30 min equilibration(42.95 +/- 2.55 and 50.39 +/- 2.42) were better (P0.05) than those of the other groups (10 min: 31.33 +/- 3.06 and 38.98 +/- 3.31; 60 min: 32.49 +/- 3.86 and 40.01 +/- 4.18; 90 min: 31.16 +/- 3.66 and 38.30 +/- 3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity.</p><p><b>CONCLUSION</b>Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol.</p>