3.The technologies of small molecule ligand-target protein interaction and high throughput ligand screening
Rui-fang DONG ; Yuan-zheng XIA ; Ling-yi KONG
Acta Pharmaceutica Sinica 2023;58(11):3242-3253
The interaction of drug and target protein is a critical part of new drug discovery. It is the premise for drugs to exert therapeutic effects by targeting specific binding sites of target proteins and thereby affecting its pharmacological activity. Currently, a variety of techniques are exploited to detect the interaction between drug ligands and target proteins. For example, cellular thermal shift assay (CETSA) and differential scanning fluorimetry (DSF) based on thermodynamics, mass spectrometry and nuclear magnetic resonance technology, etc. In addition, high-throughput ligand screening technology provides technical convenience for the search of specific ligand, and is a powerful tool to efficiently identify the interaction between drug ligand and target protein. Here, we summarize the detection techniques of interaction between small molecules and target proteins, and discuss the application of high-throughput ligand screening technology in drug research.
4.Label-free target identification for natural products based on proteomics
Rui-fang DONG ; Yuan-zheng XIA ; Ling-yi KONG
Acta Pharmaceutica Sinica 2023;58(8):2000-2015
Target identification and verification of natural products is an important and challenging work in the field of chemical biology. It is also an important job for researchers to apply chemical proteomics technology to biomedicine in order to identify target proteins of natural products. Target identification is critical to understanding its mechanisms and developing natural products as molecular probes and potential therapeutic drugs. Traditional approaches of small molecule target identification based on affinity have been shown to be successful, such as click-chemical probes, radioisotope labeling or photosensitized small-molecule probes. Nevertheless, these technologies require purified candidate target proteins, and modified small molecules with probes or linkers, such as adding agarose beads, biotin labels, fluorescent labeling or photo-affinity labeling. Many structure-activity relationship studies should be performed to ensure that the addition of small molecule labels undisturbed the original biological activity of the small molecules. Unfortunately, all these modifications are likely to alter their biological activity or binding specificity. To overcome the bottleneck of "target recognition", researchers have developed a series of new techniques for unmodified drug target identification. In this article, we reviewed the target identification techniques of natural product without structural modification in order to provide reference for the development of natural products.
5.Experience and thinking on establishing fine course of oral and maxillofacial surgery
lian, GUO ; zhi-yuan, ZHANG ; jia-wei, ZHENG ; xia, CAO ; yuan-jin, XU ; chi, YANG
Journal of Shanghai Jiaotong University(Medical Science) 2008;0(S1):-
Objective To explore the practical ways on establishing fine course of oral and maxillofacial surgery effectively. Methods Relying on the advantages of the discipline,great efforts had been made in step-by-step enhancement of the quality of teachers,teaching contents,teaching methods and administration. Results Through the establishing of fine course,we could improve the curriculum system,enhance the force of education team,and improve the quality of education. Conclusion Establishing fine course of oral and maxillofacial surgery depends on the environment of sharing educational resources,adjusting the curriculum system and establishing an excellent educational team.
6.Effect of methylprednisolone on reperfusion injury in severe uncontrolled hemorrhagic shock.
Fang XIA ; Jing-shan CAO ; Li-ying ZHAN ; Zhong-yuan XIA ; Zheng-yuan XIA ; Hai-bo HUANG
Chinese Journal of Traumatology 2003;6(6):359-362
OBJECTIVETo study the effect of methylprednisolone (MP) on reperfusion injury in severe uncontrolled hemorrhagic shock and explore the possible mechanism involved.
METHODSTwelve dogs were randomly divided into two groups, control group (Group I, n=6) and MP group (Group II, n=6). The animals were bled continuously from a femoral artery catheter to produce uncontrolled hemorrhagic shock models. Resuscitation with lactated Ringer's (LR) solution was initiated when mean arterial pressure (MAP) decreased to 20 mm Hg, and MAP was maintained at 30-40 mm Hg. MP (4 mg/kg) was injected intravenously in Group II when resuscitation began. While in Group I, normal saline (NS) was injected instead. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured before exsanguination (T(1)), when MAP decreased to 20 mm Hg (T(2)), 60 min (T(3)) and 120 min (T(4)) after resuscitation. Heart rate, MAP and cardiac output (CO) levels were recorded concomitantly.
RESULTSInfusion volume and hemorrhage volume shed from the superior mesenteric artery in Group I were higher than those in Group II (P<0.01 and P<0.05). After reperfusion, blood SOD levels decreased progressively and MDA levels increased rapidly in Group I. In Group II, blood SOD levels at T(3) and T(4) decreased as compared with that at T(1) but a stepwise increase was present. At T(4), blood SOD level was significantly higher in Group II than in Group I (Plt;0.01). At T(3) and T(4), MDA levels were markedly lower in Group II than in Group I. During reperfusion, MAP was more steady in Group II than in Group I and survival rate after 120 min (at T(4)) was higher in Group II than in Group I (P<0.05).
CONCLUSIONSMP has a protective effect on severe uncontrolled hemorrhagic shock and subsequent reperfusion injury. The mechanism mainly involves the anti-lipid peroxidation activity of MP.
Analysis of Variance ; Animals ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Female ; Lipid Peroxidation ; Male ; Methylprednisolone ; pharmacology ; Probability ; Random Allocation ; Reference Values ; Reperfusion Injury ; drug therapy ; physiopathology ; Sensitivity and Specificity ; Shock, Hemorrhagic ; drug therapy ; physiopathology ; Survival Rate
7.The imaging study on the value of ~1H-MR spectroscopy in diffuse axonal injury
Hai-Jian XIA ; Xiao-Chuan SUN ; Wen-Yuan TANG ; Lv-Ping ZHENG ;
Chinese Journal of Radiology 2001;0(04):-
Objective To investigate the value of ~1H-MRS in the diagnosis and prognosis of diffuse axonal injury(DAI).Methods A prospective imaging study was performed in 63 patients with craniocerebral injury admitted from October 2002 to April 2004.Sixty-three patients were divided into DAI group(27 cases)and Non-DAI group(36 cases)according to the result of the MRI.Then,the ratio of NAA/Cr,Cho/Cr,mINs/Cr,and GIx/Cr at basal ganglia and genu and splenium of corpus callosum was quantified using ~1H-MRS and compared between DAI group and Non-DAI group.Twenty healthy persons were served as control group.The relation between ~1H-MRS indexes and period of primary uneonciousness post-injury was analyzed.Results The results of NAA/Cr and Cho/Cr at genu and splenium of corpus callosum and basal ganglia of control group were 1.19?0.18,1.21?0.24;1.89?0.17,1.84?0.14; 1.57?0.16,1.85?0.25,which of DAI group were 0.83?0.24,2.92?0.78;1.25?0.35,2.54? 0.42;1.33?0.17,2.38?0.44,and those of Non-DAI group were 1.11?0.23,1.61?0.33;1.61? 0.22,1.93?0.26;1.49?0.23,1.89?0.29.The differences between them were statistically significant (P
8.Fasting plasma obestatin levels in patients with different glucose tolerance and its association with lipid metabolism
Pengying GU ; Dongmei KANG ; Yuan XIA ; Hui ZHENG ; Weidong WANG ; Jian ZHOU ; Cuiping ZHAO ; Yan CHEN
Chinese Journal of Geriatrics 2010;29(6):480-482
Objective To investigate whether fasting obestatin level is different in patients with impaired glucose tolerance or type 2 diabetes, and to explore the association between obestatin and lipid metabolism. Methods Eighty-four subjects without known diabetes were divided into three groups: normal glucose tolerance(NGT), impaired glucose tolerance (IGT) and type 2 diabetes (DM) Plasma obestatin levels were measured with a radioimmunoassay. The relationship between fasting obestatin levels and metabolic parameters was also analyzed. Results Fasting obestatin levels were lower in DM group [(2.82±0.78)ng/ml] and IGT group [(3.25±0.29)ng/ml] than in NGT group[(3.55±0.57) ng/ml, P<0.01]. Triglycerides and low density lipoprotein cholesterol levels gradually increased among the three groups (P<0.05). Multiple linear regression analysis revealed fasting obestatin level was independently associated with waist-to-hip ratio, triglyeride and low density lipoprotein cholesterol. The regression equation was obestatin=6.953-3.412×W/H-0.175×TG-0.123×LDL-C. Conclusions The decreased obestatin may be associated with IGR and T2DM, and obestatin level may be associated with lipid metabolism.
9.The significance of the vitamin D anti vitamin D receptor In the initial systemic lupus erythematosus patients
Yijun ZHENG ; Dawei HU ; Sheng CHEN ; Yi TAN ; Chunde BAO ; Jiajing XIA ; Wen AO ; Min YUAN
Chinese Journal of Rheumatology 2009;13(9):595-598
Objective ① To investigate the level of the vitamin D endocrine system in peripheral relationships with bone mineral density (BMD) and the disease activity respectively. Methods The level of the 25-hydroxylate vitamin D3 (25OHD3) and 1,25-dihydroxyvitamin D3 [1,25(OH)D3] in plasma from 43 SLE patients and 44 normal controls were detected by enzyme linked immunosorbent assay. Vitamin D receptor (VDR) gene expression was determinied by real-time PCR in peripheral blood. BMD measurements in the lumbar spine (L1-4) and left proximal femur (femoral neck) were performed using dual X-ray absorptiometry before treatment. The relationship between the vitamin D endocrine system and the bone mass were studied. We also discussed the relationship between the vitamin D endocrine system and the disease activity. Results The levels of 25OHD3 and 1,25 (OH)2D3 were lower in the initial SLE patients than normal controls (P<0.01, P<0.01). The expressions of VDR gene were significantly increased in initial SLE compared with normal controls (P<0.01). The initial SLE patients had significantly lower BMD values, and higher frequency of osteopenia (35%) at both sites of measurement compared with matched healthy controls (P<0.01). The initial SLE patients were divided into two groups by BMD, abnormal group and normal group. There were no differences in 25OHD3, 1,25 (OH)D3 and VDR gene expression (P0.05). There was no correlation between the vitamin D endocrine system and BMD in initial SLE patients. There was no correlation between the vitamin D endocrine system and the disease activity either. Conclusion Vitamin D endocrine system may play an important role in SLE, but the level of VDR gene is not correlated with BMD and disease activity.
10.Sanguinarine inhibits cell proliferation in MCF-7 human mammary adenocarcinoma cells
Yupan LU ; Xianzhe DONG ; Xia FENG ; Yuan HU ; Xiaoli ZHENG ; Xiaoyue GE ; Jinliang WANG ; Ping LIU
Chinese Pharmacological Bulletin 2016;32(6):858-862
Aim Toinvestigatewhetherexposureto Sanguinarine (SAN ) can inhibit cell proliferation in human mammary adenocarcinoma cells (MCF-7 ) and thepossiblemechanism.Methods WeexposedMCF-7 to anticancer compound SAN,cell viability was as-sessed by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT ) reduction assay. ROS was measured using confocal microscopy,expres-sion of caspase-3 ,caspase-8 and caspase-9 were calcu-latedusingchemiluminescencemethod.Results SAN remarkably inhibited growth of human mammary adeno-carcinoma MCF-7 cells by decreasing cell proliferation. ROS release and caspase-3,caspase-8,caspase-9 ex-pression were stimulated by SAN in MCF-7 ,and these changes were abolished by the antioxidant,N-acetyl-cysteine(NAC).Conclusion Regulationofcaspases expression and release from MCF-7 cells are possibly e-voked by SAN through reactive oxygen species.