1.Studies on Mechanisms of Synergic Action of Radix Sophorae Flavescentis and Fructus Cnidii for Treatment of Pruritus Based on Network Pharmacology
Jiali ZHONG ; Danling LIANG ; Dan LI ; Xinxin ZHOU
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(5):742-752
Objective To explore the mechanisms of synergic action of commonly-used herb pair of Radix Sophorae Flavescentis (Kushen, KS) and Fructus Cnidii (Shechuangzi, SCZ) for the treatment of pruritus. Methods We predicted and analyzed the potential targets of KS and SCZ based on network pharmacology method, and then established Chinese herbs-compound skeletons-targets networks to reveal anti-pruritus targets, predicting targets, and the interaction of KS and SCZ, as well as the components of the herb pair. The RAW264.7 inflammatory cell model was established to observe the synergistic anti-inflammatory activity of KS and SCZ. Results KS and SCZ had possible synergic action on the pruritus-related targets such as histamine receptors, cannabinoid receptors and proteinase-activated receptor 2. Additionally, KS and SCZ probably had synergic regulation of inflammation-related pathway (Toll -like receptor signaling pathway , chemokine signaling and Fc epsilon RI signaling pathway) and nerve-related pathway(neurotrophin signaling pathway) during the treatment of pruritus. Flavones from KS and coumarin of SCZ had various synergistic anti-inflammatory activities(P<0.05), indicating that they played an important role in inhibiting pruritus induced by inflammation. Conclusion The method may reveal the molecular mechanism of KS and SCZ in inhibiting pruritus, which is important for the modernization of Chinese medicine and new drug development.
2.Effect of flow shear stress on endothelial cell proliferation and c-myc expression in cerebral arteriovenous malformation
Mingguang ZHAO ; Yanbing LI ; Bochuan Lü ; Yong LIANG ; Hongli XUE ; Liping ZHAO ; Danling WANG
Chinese Journal of Tissue Engineering Research 2007;11(36):7278-7281
BACKGROUND:Shear stress can directly mediate the expression of endothelial cells, especially some cytokine genes whose codes are related to angiogenesis. Otherwise, flow shear stress of blood plays an importantly biological role in regulating vascular structure and function.OBJECTIVE: To observe the effects of laminar flow shear stress on the proliferation of vascular endothelial cells and the expression of protooncogene c-myc in human cerebral arteriovenous malformation.DESIGN: Randomized controlled study.SETTING: Department of Neurosurgery, Shenyang General Hospital of Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Neuromedical Institute, General Hospital of Shenyang Military Area Command of Chinese PLA from November 2006 to February 2007. Fresh samples of human cerebral arteriovenous malformation were derived from 20 patients who were of grade Spetzler Ⅱ -Ⅲ and received resection of human cerebral arteriovenous malformation in the Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA in 2006. All cases were diagnosed with whole-brain angiography before operation. The main reagents and equipments were detailed as follows: M199 culture media (Gilbco BRL), quality fetal bovine serum (HyClone), endothelial cell growth supplement (ECGS; Sigma, USA), CO2 incubator (Forma Scientific, USA), flow cytometry analysis of cell cycle kit (BD Company), flow cytometer (FACS Calibur, BD Company), rat-anti-human c-myc monoclonal antibody (Santa Cruz Company, USA), and reverse transcription polymerase chain reaction (RT-PCR) kit (Promega).METHODS: Tissue explants adherent method was used to culture vascular endothelial cells of human cerebral arteriovenous malformation, and then the cells were classified into 4 groups based on degree of shear stress, including control group, low shear stress group, moderate shear stress group and high shear stress group. Cultured endothelial cells of human cerebral arteriovenous malformation were put in a parallel plate flow chamber. In addition, cells in the low,moderate and high shear stress groups were stressed by low, moderate and high shear stress for 8 hours, respectively.However, shear stress in the control group was 0 Pa. Flow cytometry was used to measure proliferation index, and the expression of c-myc protein and c-myc mRNA were determined by immnocytochemistry and RT-PCR analysis respectively.MAIN OUTCOME MEASURES: Expressions of c-myc protein and c-myc mRNA and proliferation index in endothelial cells under various degrees of shear stress.RESULTS: ① Proliferation index: Proliferation index was higher in the moderate and high shear stress groups than that in the control group (P < 0.05, 0.01). ② Expression of c-myc protein: Immuneposjtjve expression of c-myc protein was gradually increased with the increase of shear stress and there were significant differences in the three shear stress groups as compared with control group (P < 0.05-0.01). ③ Expression of c-myc mRNA: Proliferation index of endothelial cells was higher in the low and moderate shear stress groups than that in the control group (P < 0.05).CONCLUSION: Flow shear stress can induce expression of c-myc and activate expression of c-myc gene based on gene transcription so as to promote the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation
3.Inhibitory effect of vascular endothelial growth factor-antisense oligonucleotide on the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation
Mingguang ZHAO ; Bochuan Lü ; Yanbin LI ; Yong LIANG ; Hongli XUE ; Danling WANG ; Liping ZHAO
Chinese Journal of Tissue Engineering Research 2007;11(32):6517-6520
BACKGROUND:Antisense gene therapy offers immense promise for the management of human cerebral arteriovenous malformation through inhibiting expression of vascular endothelial growth factor and angiogenesis in endothelial cells.OBJECTIVE: To observe the inhibitory effect of vascular endothelial growth factor-antisense oligonucleotide (VEGF-ASODN) on the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation.DESIGN: Observational contrast study.SETTING: Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Neuromedical Institute, General Hospital of Shenyang Military Area Command of Chinese PLA from August to December 2006. A total of 18 patients with human cerebral arteriovenous malformation were selected from Department of Neurosurgery, Shenyang General Hospital of Military Area Command of Chinese PLA. There were 12 males and 6 females and their mean age was 40 years. Cerebral arteriovenous malformation was classified based on Spetzler grade: grade Ⅱ (n =10) and grade Ⅲ (n=8). All cases were diagnosed with whole cerebral angiography before operation and they provided the confirmed consent. Main reagents were detailed as follows: endothelial cell growth supplements (ECGS, Sigma, USA), 391 DNA automatic synthetic device (Shanghai Shenggong Liyong Company, PE, USA), anaerobic incubator (DY-1, Zhejiang), human vascular endothelial growth factor enzyme-linked kit (TBD Company, Beijing), 96E enzyme-labeling device (ERMA, INC), cell cycle analytical reagent kit (BD Company), and flow cytometer (FACS Calibur, BD Company).METHODS: ①Experimental procedure: Tissue explants adherent method was used to culture vascular endothelial cells from human cerebral arteriovenous malformation. The third generated cells were used and randomly divided into antisense group, sense group and control group with four bottles of cells in each group. Sense and antisense phosphorothioate oligodeoxynucleotides of artificial vascular endothelial growth factor selected from the antisense group and the sense group were covered with positive liposomes, and then they were used to transfected vascular endothelial cells cultured from human cerebral arteriovenous malformation; however, cells in the control group were not dealt with any treatments. Cells in the three groups were incubated in anaerobic incubator (including 0.95 volume fraction of N2 and 0.05 volume fraction of CO2) at 37 ℃ for 2, 4 and 8 hours, respectively. ② Experimental evaluation: Cell cycles were measured, protein content of vascular endothelial growth factor was measured, and mRNA expression of vascular endothelial growth factor was also detected.MAIN OUTCOME MEASURES: Expression of mRNA and protein of vascular endothelial growth factor and proliferation exponent at different times of hypoxia.RESULTS: ①mRNA expression of vascular endothelial growth factor: At 2, 4 and 8 hours after hypoxia, mRNA expression of vascular endothelial growth factor was higher than that before hypoxia in the control group (P < 0.05);however, mRNA expression was lower in the antisense group than that in the control group (P < 0.05). ② Protein content of vascular endothelial growth factor: At 2, 4 and 8 hours after hypoxia, protein content of vascular endothelial growth factor was higher than that before hypoxia in the control group (P < 0.05); however, protein content was lower in the antisense group than that in the control group (P < 0.05). ③ Proliferation exponent: At 4 and 8 hours after hypoxia,proliferation exponent of endothelial cells cultured from human cerebral arteriovenous malformation was higher than that before hypoxia in the control group (P < 0.05); however, proliferation exponent was lower in the antisense group than that in the control group (P < 0.05).CONCLUSION: Hypoxia may induce gene expression of vascular endothelial growth factor in endothelial cells at the transcriptional level. Antisense vascular endothelial growth factor can obviously inhibit gene expression of vascular endothelial growth factor cultured from human cerebral arteriovenous malformation and proliferation under hypoxic conditions.
4.Impact factor of relationships between CT value and relative electron density for treatment planning system
Guosheng FENG ; Yuan LIANG ; Danling WU ; Yanrong HAO ; Heming LU ; Jiaxin GHEN ; Chaolong LIAO ; Ying MO ; Yihang HUANG
Chinese Journal of Radiation Oncology 2012;21(3):281-284
ObjectiveTo study the CT values of certain phantoms scanned by various CT scanners with dissimilar parameters.Methods The CT values of tissue equivalent inserts was measured in the TM164 and CIRS-062 phantom scanned by TOSHIBA AQUILIONTM,SIEMENS SOMATOMTMSENSATIONTM 64 and SIEMENS SOMATOMTM SENSATIONTM OPEN with different voltages,currents and slice thicknesses and then the corresponding CT-to-density curves was compared. Results There are no significant differences of CT values with various currents and slice thicknesses and also for low atom number materials scanned by different scanners with various tube voltages.The CT values of high atom number materials have obvious differences scanned with tube voltage,the maximum is about 400 HU.There are also significant differences between CT-density curves of two phantoms in the range from soft tissues to dense bone,the maximum is up to 500 HU.ConclusionsCT-density curves were highly affected by materials of phantoms,scanners and tube voltages.It is necessary to measure the curve with a comfortable phantom and certain scanner to assure the accuracy for dose calculation for treatment planning system.
5.Surveillance of epidemic factors of schistosomiasis in wetlands in Sichuan Province
Yong MAO ; Jia XU ; Liang XU ; Ling CHEN ; Jia-Jia WAN ; Rong-Zhi LI ; Meng TANG ; Chao-Kun ZHONG ; Fang YANG ; Hui-Rong XU ; Yu-Ling YE ; Zi-Song WU
Chinese Journal of Schistosomiasis Control 2019;31(3):307-310
Objective To investigate the epidemic factors of schistosomiasis in wetlands in Sichuan Province, so as to provide scientific evidence for the prevention and control of schistosomiasis in wetlands. Methods The artificial and natural wetlands were selected from Sichuan Province, and the relevant data regarding wetlands were collected. Routine snail survey, investigation on human morbidity due to schistosomiasis, snail diffusion experiments, questionnaire survey, determination of water infectivity and retrospective survey were conducted. Results In Sichuan Province, the mean densities of living snails were 0.003 snails/0.1 m2 and 2.033 snails/0.1 m2 in the upper and lower reaches of the Meiwan Reservoir wetlands, 0.08 snails/0.1 m2 in the Jinyan Lake of Guanghan City, 0.21 snails/0.1 m2 in Muhe River of Guanghan City, and 0.02 snails/0.1 m2 prior to the construction of Qiong-hai wetland park in Xichang City in 2015. Artificial simulation experiments showed that the largest distance of snail diffusion in water was 2 000 m. There were 8.80% (41/466) of subjects that lived neighboring wetlands, worked in wetlands and visited wet-lands having infested water contact behaviors. A total of 690 sentinel mice were assigned, and no Schistosoma japonicum infection was detected in the 677 mice dissected. Retrospective survey showed that the construction of the Meiwan Reservoir caused the spread of schistosomiasis in Dailing County, and snails were found in the ditches entering the Jinyan Lake and in Jinyan Lake areas 5 years following the construction of the Jinyan Lake in Guanghan City, with S. japonicum -infected snails detected in the ditches entering the Jinyan Lake. Conclusions O. hupensis snails are found in some wetlands in Sichuan Province. Protection of wetlands and snail control with environmental improvements are recommended for the prevention of snail importation in natural wetlands, while in artificial wetlands, thorough snail control is recommended during the construction of the wetlands because of the likelihood of snail importation via water systems. In addition, both natural and artificial wetlands require long-term systematic surveillance of schistosomiasis.