1.Cloning, expression and bioactivity analysis of human granulysin
Wanxia WANG ; Xi LAN ; Xianghong XU ; Jun JU ; Jixing LIU
Basic & Clinical Medicine 2010;30(1):75-79
Objective To obtain recombinant human granulysin using prokaryotic expression system. Methods Total RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence, pET-GN-LY9K and pET-GNLY15K were transferred to E. Coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot. The bioactivity of granulysin fusion protein was measured by MTT assay. Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed. The corresponding protein was highly expressed in E. Coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner, while GN-LY15K had little effect on the growth of A549. Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system, which might be helpful for the further study of granulysin.
2.A case-control study about the preventive effect of magnesium sulfate on hypertention caused by ACTH in the treatment of infantile spasms
Xiuguo LIU ; Jiuwei LI ; Xiuyu SHI ; Jun JU ; Liping ZOU
Journal of Clinical Pediatrics 2014;(11):1013-1015
Objective To examine the preventive effect of magnesium sulfate on hypertension caused by ACTH in the treatment of infantile spasms (IS). Methods 46 children diagnosed as IS were recruited from two hospitals during May, 2011 to October, 2013.23 patients in group A (treatment group) were treated with magnesium sulfate and ACTH in hospital A;another 23 cases in group B (control group) were treated with ACTH only in hospital B. The therapy course was 2 weeks. Results Hyperten-sion was not observed in the treatment group, while 6 children were observed with hypertension in the control group. There was signiifcant difference between the two groups (P<0.05). Conclusions Magnesium sulfate could prevent the incidence of hyper-tension in the treatment of IS with ACTH, and beneift the completion of treatment course.
3.Cloning,expression and bioactivity analysis of human granulysin
Wanxia WANG ; Xi LAN ; Xianghong XU ; Jun JU ; Jixing LIU
Basic & Clinical Medicine 2006;0(01):-
Objective To obtain recombinant human granulysin using prokaryotic expression system.MethodsTotal RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence,pET-GNLY9K and pET-GNLY15K were transferred to E. coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot.The bioactivity of granulysin fusion protein was measured by MTT assay.Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed.The corresponding protein was highly expressed in E.coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner,while GNLY15K had little effect on the growth of A549.Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system,which might be helpful for the further study of granulysin.
4.Effect of Circadian Rhythm on Hypnotic Median Effective Dose of Ketamine
feng-ai, LIU ; jin-nan, YANG ; ti-jun, DAI ; ju-yuan, LIU ; tai-zhen, CUI
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To observe the effect of circadian rhythm on hypnotic median effective dose( ED50) of ketamine. Methods Sixty mice were randomly divided into 4 groups which had 15 mice in each group. They were intraperitoneally injected with ketamine at different times of 2 Am,8 Am,2 Pm and 8 Pm, respectively. Righting reflex was recorded and the value of ED50 was measured with sequential experimental method. Results The hypnotic ED50 of ketamine at 2 Am was(54.57?0.82) mg/kg, with 95% confidence limit of ED50 38.06-78.22 mg/kg;ED50 was(49. 27?0. 12) mg/kg at 8 Am, with 95% confidence limit of ED50 40. 21-60. 37 mg/kg;ED50 at 2 Pm was (42.28?0.21) mg/kg, with 95% confidence limit 37.35 - 47 83 mg/kg;and ED50 at 8 Pm was(57.42?0.14) mg/kg, with 95% confidence limit 37.51-73 72 mg/kg,respectively. The ED50 were significant different at 2 Pm and 8 Pm. However, there were no significant difference in ED50 value among other groups. Conclusion The hypnotic effect of ketamine has circadian rhythm - dependent.
5.Pharmacokinetic study of asiaticoside in rat plasma by UPLC-MS
Zixiu LIU ; Shijia LIU ; Wenzheng JU ; Jun ZHANG ; Min CHEN ; Hengshan TAN
Chinese Pharmacological Bulletin 2010;26(4):543-546
Aim To establish an UPLC-ESI-MS method for determination of asiaticoside and investigate its application to pharmacokinetic study in rats.Methods Eight rats were given 40 mg·kg~(-1) asiaticoside iv respectively.Drug plasma concentration was determined by UPLC-ESI-MS.Pharmacokinetic parameters were evaluated.Results Calibration curves were linear over 0.038~7.6 mg·L~(-1) and LLOQ was 38 μg·L~(-1),the recoveries of asiaticoside from plasma were larger than 95%,and RSD of inter-day and intra-day assay were below 10%.After iv administration of 40 mg·kg~(-1) asiaticoside,the pharmacokinetic parameters of AUC(0-t),T(1)/(2)β,CL,Vd were (81 443.67±57 156.81) μg·L~(-1)·min~(-1),(23.44±9.60) min,(0.19±0.07) L·min~(-1)·kg~(-1),(8.92±6.68) L·kg~(-1),respectively.Conclusion The method described in this report was sensitive and specific,and suitable for pharmacokinetic studies of asiaticoside in rats.
6.Exploring relationship between traditional effects of traditional Chinese medicine and modern pharmacological activities by "co-effect compounds".
Hai-Bo LIU ; Ai-Jun LU ; Bing LIU ; Jia-Ju ZHOU
China Journal of Chinese Materia Medica 2005;30(1):75-78
The compound that distributes in the herbs with one common effect was named as "co-effect compound" (CEC). The CECs of three traditional Chinese medicine(TCM) effects, purgative, relieving pain and clearing heat, had been found and studied. A strong corresponding relationship was found between the pharmacological activities of CECs and the TCM effect they belong to. The study shows that it may be a feasible method to connect traditional effect of TCM with modem pharmacological activity.
Anthraquinones
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isolation & purification
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pharmacology
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Anti-Inflammatory Agents, Non-Steroidal
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pharmacology
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Cathartics
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pharmacology
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Drugs, Chinese Herbal
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isolation & purification
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pharmacology
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Flavonoids
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isolation & purification
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pharmacology
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Medicine, Chinese Traditional
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Plants, Medicinal
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chemistry
7.Acute toxicity of meso-2,3-dimercaptosuccinic acid on mice
Shi-Jun LIU ; Xue-Hai JU ; Ke-Ming SUN ; Zhi-Jie CHU ; Xian-Yong LIU ; Min ZHANG ; Jun-Qing GUO ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(08):-
Objective To explore acute toxicity of succimer on mice.Methods Twenty Kunming mice(10 males and 10 females) weighting approximately (21.2?2.3)g were acclimatized for 3 days prior to dosing,then were divided into control group and experiment group with 10 mice in each group according to body weight.Fasted for 12 hours,the mice in experiment group received intragastric administration of 160mg DMSA in deionized water in 24 hours,and the control group received the same volume of deionized water,and then they were observed for 7 days.Blood was collected into heparinized-tubes by removal of eyeball.All mice were sacrificed and brain,heart,liver and kidney were removed and washed with normal saline.The activity or amount of BUN,Scr,AST,ALT,SOD, GSH-PX and MDA were analyzed.Results (1)Given 160rag DMSA in 24 hours,gastrointestinal symptoms were main side effects.During the observation,experiment group lost weight due to the decrease of food-intake ,and some mice had slight hydroabdomen.(2)High dose of DMSA caused a significant inhibition of GSH-PX(P0.05).The hepatic cell was damaged accord- ing to the significant raise of MDA in liver(P0.05),which was related to acute toxicity on liver.Conclusion Succimer could inhibit the antioxidarrt systems and could do damage to liver and kidney.
8.Abnormal rates of indexes and influencing factors related to chronic diseases of workers in Hangzhou enterprises.
Xian-fu MA ; Qing-min LIU ; Yan-ju REN ; Jun LV ; Li-ming LI
Chinese Journal of Epidemiology 2013;34(10):1049-1050
Adult
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China
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epidemiology
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Chronic Disease
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epidemiology
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Dyslipidemias
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epidemiology
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Female
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Health Status
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Humans
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Hypertension
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epidemiology
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Male
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Middle Aged
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Obesity
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epidemiology
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Smoking
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epidemiology
9.Effect of TGF-β1 and IL-1β expression in serum on acute radiation-induced heart disease
Yi WANG ; Jun WANG ; Yunjie CHENG ; Qing LIU ; Ping ZHANG ; Fengpeng WU ; Xin WAN ; Ju ZHANG
Chinese Journal of Radiological Medicine and Protection 2012;32(5):488-491
Objective To evaluate the effect of TGF-β1 and IL-1β expression in serum on acute radiation-induced heart disease (RIHD) in patients with thoracic tumors.Methods Three-dimensional conformal radiotherapy (3D-CRT) or intensity modulated radiotherapy (IMRT) was delivered at 1.8-2.0 Gy,5 times per week to a total dose of 50-66 Gy to 44 patients with lung cancer and 10 patients with esophagus cancer.The target and organs at risk dose distribution were analyzed by 3-dimensiond treatment planning system.The expressions of TGF-β1 and IL-1β in serum were detected by enzyme linked immunosorbent assay before and at the end of the irradiation.The cardiac injury was evaluated by detecting the cmyocardium creatase,cardiac troponin I (cTnI),electrocardiogram and cardiac function before and at the end of the irradiation within 90 d.The acute RIHD was evaluated by the Common Terminology Criteria V 3.0 (NCI-CTCAE 3.0).The expressions of TGF-β1 and IL-1β in the serum of RIHD patients with thoracic tumors were analyzed.Results The expression of TGF-β1 in serum was (888.4 ± 41.1) μg/L before the irradiation and approached to (926.1 ± 23.1) μg/L at the end of the radiotherapy.The expression level of TGF-β1 in the serum of acute RIDH group was (900.6 ± 34.5) μg/L,higher than that of normal group [(865.7 ±47.0) μg/L,t =-2.646,P <0.05)].The acute RIDH was correlated with the expression level of TGF-β1 before irradiation and the difference before and at the end of irradiation (r =0.378,0.311,P <0.05).The IL-1β expression had no significant difference before and after irradiation.The expression of TGF-β1 in serum before and at the end of irradiation had positive correlation with the expression of IL-1β at the end of the irradiation (r =0.416,0.389,P < 0.05).Conclusions The expression of TGF-β1 in the serum of patients with thoracic tumor increases after irradiation and correlated with the acute RIHD,but the expression of IL-1β in serum has no relationship with RIHD.TGF-β1 could induce the expression of IL-1β at the end of the irradiation.
10.Pharmacokinetic study of scutellarin in healthy volunteers by enzymatic hydrolysis and LC-MS-MS analysis
Jihong CHU ; Jun ZHANG ; Changyin LI ; Chong ZOU ; Fang LIU ; Wenzheng JU
Chinese Pharmacological Bulletin 2015;(1):108-112
Aim To establish a combined method ofβ-glucuronidase hydrolysis and LC-MS-MS analysis for the determination of scutellarein in human plasma, and investigate the pharmacokinetics of scutellarin prepara-tion in healthy male volunteers. Methods Plasma samples were prepared by enzymolysis with β-glucu-ronidase and protein precipitation with methanol. The analytes scutellarein and quercetin ( IS ) were separa-ted on an Agilent ZORBAX SB C18 column ( 2. 1 mm × 150 mm, 5 μm) with the mobile phases consisting of acetonitrile, methanol and water. Multiple reaction monitoring ( MRM) on MS was used to monitor precur-sor to produce ion transitions of m/z 285. 0→136. 8 for scutellarein and m/z 301. 1→120. 8 for IS. After method validation, this method was applied to deter-mine the plasma concentration of scutellarein in 12 male volunteers following single oral administration of 120 mg scutellarin preparation. Drug And Statistic soft-ware (1. 0) was used to process data and the pharma-cokinetic parameters were calculated. Results The assay was validated with linear range of 4 . 01-513. 38μg · L-1 for scutellarein. The intra- and inter-batch precisions ( RSD%) were within 7. 22%. The absolute recoveries were more than 84. 23%. The pharmacoki-netic parameters after a single dose were as follows:Cmax (μg · L-1 ): 159. 97 ± 58. 14; AUC(0-19) (μg · L-1·h):1151. 37 ±279. 80; AUC(0-∞)(μg·L-1· h):1194. 13 ± 264. 51; Tmax ( h):6. 33 ± 1. 67; T1/2 (h):2. 83 ± 0. 60. Conclusion The assay method is proved to be sensitive, accurate and convenient. It can be successfully applied to a pharmacokinetic study of scutellarin in healthy male volunteers.