1.Establishment of the method to evaluate cardiac toxicity by real-time cell analysis system on human embryonic stem cells
Qi ZHAO ; Xijie WANG ; Shuyan WANG ; Jing MA
Chinese Pharmacological Bulletin 2016;(1):138-143
Aim To establish an in vitro early drug cardiac tox-icity evaluation method by human embryonic stem cells derived cardiomyocytes ( hESC-CM) and real-time cell analysis Cardio (RTCA Cardio) system. Method The hESC-CM were cultured at RTCA Cardio E-Plate 96. Impedance signals from hESC-CM were analyzed for beating rate, contraction amplitude and beating rhythm irregularity to determine the optimum inoculation density and detection duration. Based on this, we used 0. 1 % DMSO to be the solvent and quinidine (0. 2, 0. 78, 3. 13, 12. 5, 50 and 100 μmol·L - 1 ) known as affecting cardiac activity to validate this method. Result The results revealed no significant changes in the cell index (CI), transient pulse patterns, beating rate and amplitude of hESC-CM. Quinidine will affect the CI and transi-ent pulse patterns of hESC-CM and decrease the beating rate and amplitude of hESC-CM when its concentration ≥3. 13 μmol · L - 1 . And this effect is concentration-dependent, the higher the concentration,the more time they need to recover beating and the more significant the beating rate and amplitude inhibition of quinidine on hESC-CM. Conclusion The method established by hESC-CM and RTCA Cardio system can detect the effect of quinidine on the contraction of hESC-CM, and this indicates that this method has the potential to be an attractive high-throughput tool for screening potential drugs in early evaluation of drug car-diotoxicity.
2.Application of human embryonic stem cells in study of drug-induced cardiotoxicity and hepatotoxicity
Shuyan WANG ; Xijie WANG ; Xiaomin HU ; Hing MA
Chinese Journal of Pharmacology and Toxicology 2014;(4):612-617
Human embryonic stem ceIIs(hESC),characterized by unique capacities of seIf-renewaI and differentiation into cardiomyocytes and hepatocytes,can be used in new drug screening and drug safety evaIuation processes. Cardiotoxicity and hepatotoxicity are major obstacIes to deveIopment and marketing of new drugs. hESC-derived cardiomyocytes and hepatocytes have structuraI and functionaI characteristics,which can be used for cardiotoxicity and hepatotoxicity testing in vitroand for buiIding a drug safety evaIuation system invitrothat has the advantage of short experiment cycIes,smaII dose,Iow cost and few species differences. hESC-derived cardiomyocytes and hepatocytes have broad prospects of appIication in toxicoIogy.
3.Determination of Arsenic and Copper in Chinese Patent Medicines by Microwave-digestion Atomic Absorption Spectrometry
Lijuan ZHANG ; Qun MA ; Jing WANG ; Xuexin GU ; Shuyan WANG
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(05):-
Objective To establish a method for the determination of arsenic and copper in Chinese patent medicines.Methods Digested by microwave,the content of arsenic in Liuwei Dihuang Pill (prepared with water and honey) was measured by sequence flowing injection-hydride generation-atomic absorption spectrometry and content of copper by flame atomic absorption spectrometry. Effects of factors,including digestion temperature,smashing granularity of samples,proportion of digestive ingredients,volume of digestive agents,digestion procedure and digestion time,on the content determination were studied.Results Recoveries of arsenic and copper were 100.8%~114.3%and 94.4%~110.2%respectively under optimum conditions.Conclusion This method is simple and effective for the content determination of heavy mental in Chinese patent medicines.
4.A real-time cell analysis system for evaluating anti-arrhythmic drugs by monitoring the growth and beating of primary neonatal rat cardiomyocytes
Shuyan WANG ; Xijie WANG ; Kang JLN ; Taotao HUL ; Jing MA
Chinese Journal of Pharmacology and Toxicology 2014;(6):837-843
OBJECTlVE To establish a real-time cell analysis system ( RTCA) for early drug car-diotoxicity evaluation. METHODS An in vitro drug cardiotoxicity evaluation method was established using RTCA Cardio system and primary cultured cardiomyocytes of neonatal rats. The beating rate, am-plitude and beating rhythm irregularity ( BRl ) of cardiomyocytes were observered after antiarrhythmic drugs, such as quinidine and lidocaine were added, to assess the effect of the above method on cardio-toxicity evaluation. RESULTS RTCA Cardo E-Plate 96 was inoculated with primary cultured cardiomyo-cytes that began to beat after 24 h and beat regularly after 48 h. The stable beating was maintained for a minimum of three days. The beating of cardiomyocytes decreased rapidly from 155±5 to 0 after incuba-tion with quinidine. The beating recovered gradually after 6 h. Quinidine at 3.1μmol·L-1 caused the beat-ing rate to return to 124±16. Quinidine allowed the beating rate to return to normal when the concentra-tion was less than 100.0μmol·L-1 . The beating rate, amplitude and BRl of cardiomyocytes changed in a concentration-dependent manner when incubating with lidocaine. The higher the concentration, the more significant the inhibition of lidocaine on cardiomyocytes. CONCLUSlON The cardiotoxicity of quinidine and lidocaine can be detected accurately using RTCA Cardio system, suggesting that this system can be used in early evaluation of drug cardiotoxicity.
5.Influence of Qingzhifugan Pellet on fatty liver animal model with chronic hyperlipemia and alcoholized hepatic damage
Bo YANG ; Shuyan WANG ; Fangzhou LIU ; Xiuyun MA ; Guolong ZHAO
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(02):-
Objective:To observe influence of qingzhifugan pellet on mixed-type animal model of fatty liver with chronic hyperlipemia and alcoholized hepatic damage. Methods:The mixed-type animal model of fatty liver was established by administering with high lipids and alcohol feeds. Therapeutic experiments were conducted with a self-made new drug the qingzhifugan pellet. The efficacy of the tested drug was evaluated comprehensively by such measures as sero-level of lipids and ferments,as well as the histological pathology of liver. Results:It was shown that the tested drug can decrease the sero-level of lipids,promote the metabolism of lipids in liver,recover the alcoholized hepatic damage,and relieve the liver fat denaturation. Conclusion:It suggested that the tested drug possessed certain effect for treating or improving chronic hyperlipemia and alcoholized hepatic damage in the animal model.
6.Influence of Transdermal Electric Stimulation of Neiguan (PC 6) On TCD in Patients with Acute Migraine
Debao YE ; Jianqiao FANG ; Yinghui SONG ; Shuyan ZHANG ; Zeyun MA ; Weiai LIU
Journal of Acupuncture and Tuina Science 2006;4(1):22-24
Purpose: In order to explore the clinical mechanism of Neiguan (PC 6) in the treatment of acute seizure of migraine and the relationship between Neiguan (PC 6) and cerebral blood flow. Methods: Clinically, 30 cases of the patients with acute seizure of migraine (ascending phase) were selected, in TCD abnormality, and divided randomly into the treatment group and control group, 15 cases in each group. In acute seizure of migraine, Neiguan (PC 6)was punctured with electric stimulation on the healthy side of the patients in the treatment group and Xuehai (SP 10) was punctured with electric stimulation on the healthy side of the patients in the control group. TCD was determined in the patients 30 min before and after the treatment. Results and conclusion: After transdermal electric stimulation on Neiguan (PC 6),the velocity of intracranial blood flow was obviously slowed down, indicating that transdermal electric stimulation on Neiguan (PC 6) can effectively relieve the spasm of intracranial blood vessels and also shows its relative specificity.
7.Study on the effects of mM-CSF in the functional polarization of macrophages
Linlin XU ; Guoguang ZHENG ; Cuihua MA ; Hairong JIA ; Jinghui ZHONG ; Shuyan LIU ; Yongmin LIN
Journal of Leukemia & Lymphoma 2010;19(7):388-390
Objective To investigate the anti-inflammatory/immune modulatory effects of high-expressing membrane bound M-CSF-hematopoietic malignant cells on macrophages. Methods After coculturing RAW264.7 and murine macrophage cell line with Namalwa-M, a cell line stably expressing mM-CSF, and companing with Nainalwa-V, a cell line stably transfected with the empty vector as the control, flow cytometry was used to detect the expression of the marker of alternatively activated macrophages, CD206, and intracellular expression of IL-10, IL-12, IL-6 and TNF-α to study the immunophenotype of RAW264.7; phagocytic assays to investigate their functional activity in vitro. Results RAW264.7 cocultured with Namalwa-M consistently showed high-level expression of CD206, which indicated activities of these macrophage cells were increased. Furthermore, these RAW264.7 expressing high levels of IL-10, TNF-a and low levels of IL-12, IL-6, as determined by intracellular staining were suggested that the phagocytic activity was increased. Functionally, RAW264.7 cocultured with Namalwa-M showed a higher level of phagocytic activity. Conclusion Macrophage generated in vitro after cocultured with mM-CSF-expressing hematopoietic malignant cell line could be transformed into abnormal macrophage with an immunophenotype defined as IL-10-high, IL-12-low, IL-6-low and TNF-α-high.
8.Establishment of reproducible xenotransplantation model of T cell acute lymphoblastic leukemia in NOD/SCID mice.
Di, WANG ; Na, WANG ; Yan, ZHANG ; Shuyan, MA ; Zhe, GENG ; Pengfei, ZHOU ; Jianfeng, ZHOU ; Liang, HUANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(4):511-6
T cell acute lymphoblastic leukemia (T-ALL) is an aggressive leukemia. However the poor prognosis and low morbidity restrict further analysis of the disease. Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches. In this study, we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein. Four of the 9 patients and the cell line were successfully engrafted. Flow cytometry detected high percentage of human CD45(+) cells in recipient mice. Immunohistochemistry showed infiltration of human CD45(+) cells in different organs. Serial transplantation was also achieved. In vivo drug treatment showed that dexamethasone could extend survival, which was consistent with clinical observation. These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice, which recapitulated the characteristics of original disease.
9.Influence of hydrotherapy combined with breathing training on lung function and activity ability of patients with thoracolumbar spinal cord injury
Deng YANG ; Ying CHEN ; Ye WANG ; Weitao WANG ; Cheng YANG ; Qiang MA ; Pengsheng HUI ; Shuyan QIE
Clinical Medicine of China 2021;37(3):269-274
Objective:To explore the effect of hydrotherapy combined with breathing training on lung function and mobility of patients with thoracolumbar spinal cord injury.Methods:A total of 80 patients with thoracolumbar spinal cord injury admitted to Beijing Rehabilitation Hospital Affiliated to Capital Medical University from April 2018 to March 2020 were selected as observation objects.A prospective cohort study was conducted and randomly divided into observation group and control group with 40 cases in each group.The control group was given routine rehabilitation therapy combined with respiratory training.On the basis of the control group, the observation group was treated with water therapy.The indexes of lung function, motor function, lower limb muscle tension, function evaluation and activities of daily living were compared between the two groups.Results:After intervention, the observation group′s forced vital capacity (FVC) was (3.86±0.82) L, the forced expiratory volume in the first second (FEV1) was (3.76±0.68) L, the maximum ventilation (MVV) was (102.34±10.38) L/min, the maximum suction pressure (MIP) is (50.36±4.62) cmH 2O; the control group FVC was (3.41±0.76) L, and FEV1 was ( 3.35±0.63) L, MVV was (90.67±11.68) L/min, MIP was (44.38±4.85) cmH 2O, the difference between the two groups is statistically significant ( t=2.546, 2.797, 4.723, 5.646, respectively, all P<0.05). After the intervention, the motor function score of the american spinal injury association (ASIA) of the observation group was (58.62±7.56) points, and the modified ashworth scale (MAS) score was (2.74±0.89) points; The ASIA motor function score of the control group was (42.24±6.40) points, and the MAS score was (3.36±0.94) points.The difference between the two groups was statistically significant ( t=10.459 and -3.029, respectively, all P<0.05). After intervention, the observation group′s spinal cord independence measure (SCIM III) score was (75.33±10.72) points, and the modified barthel index (MBI) was (66.64±6.34) points; the SCIM III score of the control group was (68.34±9.55) points, and the MBI score was (57.52±6.77) points, the difference between the two groups was statistically significant ( t=3.079 and 6.219, respectively, all P<0.05). Conclusion:Hydrotherapy combined with breathing training can significantly improve lung function and respiratory muscle strength in patients with thoracolumbar spinal cord injury, and improve motor function and ability of daily living.
10.Effect of Isometric Contraction on Sympathetic Nervous Symptoms Followed Subaxial Cervical Instability
Shuyan QIE ; Xialin ZHANG ; Yu PAN ; Quansheng MA ; Lihua ZHANG ; Yonghong YU
Chinese Journal of Rehabilitation Theory and Practice 2013;19(3):269-271
Objective To study the effect of isometric contraction on subaxial cervical instability with sympathetic nervous symptoms.Methods 41 subaxial cervical instability patients with sympathetic nervous symptoms were assigned to experimental group (n=21) and control group (n=20). Both groups received manipulation therapy. Experimental group received isometric contraction in addition. They were assessed with Visual Analogue Scale (VAS) and heart rate variability (HRV), including total power (TP), low frequency power (LF), high frequency power (HF), and LF/HF before and 3 weeks after treatment. Results The scores of VAS, LF and LF/HF decreased in both groups after treatment (P<0.05), which decreased more in the experiement group than in the control group (P<0.05). Conclusion Isometric contraction can release the sympathetic nervous symptoms post subaxial cervical instability.