1.Anti-liver fibrosis activities of the extracellular domain of transforming growth factor beta type II receptor fusion protein in vivo
Yanfei CAI ; Aini WAN ; Yun CHEN ; Jian JIN
Journal of China Pharmaceutical University 2019;50(2):246-252
This study is performed to analyze the anti-liver fibrosis effect of the fusion protein of human serum albumin and extracellular domain of transforming growth factor beta type II receptor(eTGFBR2)in vivo to looking for the more stable anti-liver fibrosis drug. The mice model of liver fibrosis was constructed by CCl4 induction and the following groups are included in the study: the control group, CCl4 model group, the positive control group, eTGFBR2 treatment group, HSA-eTGFBR2 treatment group, and HSA group. Hematoxylin eosin staining, serum liver function index detection, and western blot are used to identify the anti-liver fibrosis activities. The results showed that: (1)CCl4 caused liver structure disorder, hepatocellular necrosis, collagen fibers proliferation, and induced liver fibrosis at last; (2)HSA-eTGFBR2 and its monomer drug improved the symptoms of liver fibrosis significantly, as well as reduced the damage of liver cells and collagen deposition, and recovered the liver basic structure to normal. Both of HSA-eTGFBR2 and its monomer drug improved liver function and reduced the expression level of liver fibrosis marker α-SMA and COL I. Moreover, the anti-liver fibrosis effect of the fusion protein is comparable to the monomer drug. In contrast, the albumin had no effect on therapeutic effect; (3)Reducing the injection frequency of HSA-eTGFBR2 achieved the comparable effects to the monomer drug with the normal injection frequency. In summary, the fusion protein HSA-eTGFBR2 has good anti-liver fibrosis effect. In addition, reducing the injection frequency of the fusion protein could also achieve the comparable treatment with the monomer drug, indicating that the fusion protein is stable and has longer half-lives and then a relatively positive application prospect in future.
2.Superconducting MRI Signal Intensity in Pallidum in Neonatal Hyperbilirubinemia
Lijuan CHEN ; Xiaoming WANG ; Yuzhen WAN ; Weihai LI ; Yonggeng JIA ; Caihong YUN ; Tianhu CHEN
Chinese Journal of Rehabilitation Theory and Practice 2016;22(7):838-840
Objective To explore the relationship between MRI signal intensity in pallidum and levels of total bilirubin in neonatal hy-perbilirubinemia. Methods From July, 2014 to October, 2015, sixty neonates were divided into three groups according to the levels of total serum bilirubin (TSB), that were group I (TSB 17.1~34.2μmol/L, n=16), group II (TSB>34.2~340μmol/L, n=34), and group III (TSB>340μmol/L, n=10). They were screened with 3.0 T MRI, and the T1WI signal intensity of bilateral pallidum was measured. Results The bi-lateral signal intensity was higher in group III than in group II and group I. There was positive correlation between signal intensity and TSB levels. Conclusion The MRI signal intensity in pallidum may help for diagnosis of neonatal bilirubin encephalopathy.
3.Construction of Lmdd-LMP2A, an attenuated Listeria vaccine strain expressing the Epstein-Barr vi-rus latent membrane protein 2A (EBV-LMP2A) and evaluation of its anti-tumor effects against na-sopharyngeal carcinoma
Wei ZHAO ; Xiaoqin CHEN ; Xin WAN ; Ci CHENG ; Zhe LIN ; Dongju FENG ; Kun YAO ; Yun CHEN
Chinese Journal of Microbiology and Immunology 2015;(3):207-212
Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.
4.Effects of chronic exposure to sub-anesthetic concentrations of sevoflurane on memory function and homeostasis of mice
Liufang CHEN ; Quanyuan FU ; Jing WAN ; Zongze ZHANG ; Yanlin WANG ; Yun WU
Chinese Journal of Anesthesiology 2014;34(4):451-453
Objective To evaluate the effects of chronic exposure to sub-anesthetic concentrations of sevoflurane on memory function and homeostasis of mice.Methods Thirty-six healthy male Kunming mice,aged 40 days,weighing 25-30 g,were randomly assigned into 3 groups (n =12 each):control group (group C),0.1% sevoflurane group (group L) and 0.5 % sevoflurane group (group H).The mice inhaled 0.1% (group L) or 0.5% sevoflurane (group H) between 18:00-6:00 (the next day) every night for 30 days.Water maze test was performed at 27-30 days of inhalation.Blood samples were collected from the left ventricle for blood gas analysis and for determination of blood electrolytes.Results There was no significant difference in swimming time in Water maze test,number of errors and blood gas analysis and blood electrolytes.Conclusion Chronic exposure to subanesthetic concentrations of sevoflurane has no significant effects on memory function and homeostasis of mice.
5.Liver fibrosis and stem cell therapy
Dongsheng XU ; Aini WAN ; Yun CHEN ; Yang HE ; Jianfeng YANG ; Jian JIN
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(3):403-406
Liver fibrosis is a common chronic liver disease, which is a stress response process of liver cells affected by one or more pathogenies for long term or repeatedly. During the fibrosis process, massive accumulated extracellular matrix can form scar tissue, which results in liver dysfunction or failure and seriously endangers the health of people. According to many independent reports, stem cell therapy can facilitate the alleviation of liver fibrosis. During the stem cell therapy, stem cells migrate to the injury site of liver and alleviate the liver fibrosis by improving the microenvironment of the scar area via paracrine way. This article reviews the formation, treatment, and stem cell therapy of liver fibrosis.
7.Fermentation of Bacillus subtilis ge25 strain and preliminary study on its antagonistic substances.
Chen-Yun HU ; Yong LI ; Min LIU ; Wan-Long DING ; Min-Jian QIN
China Journal of Chinese Materia Medica 2014;39(14):2624-2628
Panax ginseng is one of the most important traditional Chinese herbal medicine, soil borne diseases influenced the yield and quality severely. In our previous work, endophytic Bacillus subtilis ge25 strain was isolated from ginseng root, and which showed significant antagonistic activity against several most destructive ginseng phytopathogens. In the present work, crude protein and lipopeptid extracts were prepared from LB and Landy supernate by salting out, acid precipitation methods respectively. The antagonistic activity of crude extracts and stability to temperature and protease digestion were examined by ginseng phytopathogen Alternaria panax. Results showed that, the antagonistic activity of crude protein extracts from LB culture was complete and partially lost when treated by high temperature and proteinase K. However, crude lipopeptid from Landy culture showed significant stabile antagonistic activity to them. Acid-hydrolyzation and TLC-bioautography analysis showed, that the crude lipopeptide contained at least one cyclic lipopeptide. In consideration of the stability and perfect antagonistic activity of ge25, further researches will promote the biocontrol of ginseng diseases in the field.
Alternaria
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drug effects
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physiology
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Bacillus subtilis
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metabolism
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physiology
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Bacterial Proteins
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isolation & purification
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metabolism
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pharmacology
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Endopeptidase K
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metabolism
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Endophytes
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metabolism
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physiology
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Fermentation
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Lipopeptides
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isolation & purification
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pharmacology
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Panax
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microbiology
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Plant Roots
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microbiology
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Temperature
8.Anti-liver fibrosis activities of human insulin-like growth factor-1 in vitro
Aini WAN ; Dongsheng XU ; Yanfei CAI ; Yun CHEN ; Jian JIN ; Huazhong LI
Journal of China Pharmaceutical University 2017;48(4):476-482
This study was focus on investigating the anti-liver fibrosis effects of insulin-like growth factor-1 (IGF-1) in vitro.The effects of IGF-1 on human liver L-02 cell viability and cell cycle were observed.CC14-induced L-02 cell injury was set up to detect the anti-apoptotic activity of insulin-like growth factor-1 (IGF-1).Transforming growth factor β1 (TGF-β1) induced hepatic stellate cell line (HSC-T6) were used as a liver fibrosis model in vitro to analyze the effects of IGF-1 on the expression of liver fibrosis proteins and intracellular TGF-β1/Smad signaling pathway in HSC-T6 cells.The results showed that IGF-1 could relieve the growth inhibition effects of TGF-β1 on L-02 cells,increase the viability of L-02 cells injured by CCl4,decrease the expression of liver fibrosis proteins,and inhibit the TGF-β1/Smad signaling pathway by inhibiting the phosphorylation of Smad3.Our study suggested that IGF-1 exerted anti-liver fibrosis effects by stimulating L-02 cells proliferation,reducing cell damage and inhibiting ECM accumulation via interfering TGF-β1/Smad signaling pathway.
9.Findings of MRI in Cervical Spinal Cord Injury without Fracture and Dislocation and Correlation between Apparent Diffusion Coefficient of Foci and Severity of Injuries
Xiaoming WANG ; Lijuan CHEN ; Yonggeng JIA ; Yuzhen WAN ; Caihong YUN ; Liang ZHOU ; Shengwei WANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(4):394-396
Objective To study the MRI of cervical spinal cord injury without fracture and dislocation and explore of the apparent diffusion coefficient (ADC) of injured site related to the severity of injury. Methods 46 patients with cervical spinal cord injury without fracture and dislocation and 20 healthy controls were scaned with routine MRI and diffusion weighted imaging. The ADC value of site of injury and grades of Frankel's classification were analyze with the correlation. Results There were 22 cases with spinal cord edema, 8 cases with intramedullary hemorrhage, 14 cases with edema and hemorrhage, 2 cases without abnormal finding. The ADC of controls and patients were (1.05±0.12)×10-3 mm2/s, (1.21±0.23)×10-3 mm2/s (t=0.704, P<0.05). The ADC values positively correlated with the grades of Frankel's classification (r=0.407, P<0.05). Conclusion MRI may help to find the cervical cord injury in those without fracture and dislocation, and the ADC may be resposible to the severity of injury.
10.Study on Effects of Saikosaponin-d on TPA,PAI,MDA and NO in Rats with Liver Fibrosis
Fang WAN ; Jingzhen GUO ; Xin LI ; Ping LI ; Yuwu CHEN ; Keming LI ; Chen WANG ; Yun ZHANG ; Jing LI ; Yan LIU
China Pharmacy 2005;0(24):-
OBJECTIVE:To investigate the effects of saikosaponin-d(SSd) on tissue plasminogen activator(TPA),plasminogen activator inhibitor(PAI),malonaldehyde(MDA) and NO in rats with liver fibrosis induced by dimethylnitrosamine(DMN).METHODS:Eighteen SD healthy rats were randomized to control group(NS ip qd for 4 weeks),model group(10mg? kg-1 DMN ip 3 times per week for 4 weeks) and SSd-treated group(10mg? kg-1 DMN ip 3 times per week + SSd 1.8 mg? kg-1 ip for 28 consecutive days).All rats were killed 1h after the last time of administration,blood samples were taken from abdominal aorta and liver samples were taken for the observation of pathology and detection of indices of TPA,PAI,MDA and NO etc.RESULTS:SSd could lessen the degree of liver fibrosis and improve the fibrinolytic activities of TPA and PAI,meanwhile,it showed clearance effect on MDA and marked protective effect on hepatic cells.There were significant differences between SSd-treated group and the model group.CONCLUSION:SSd exhibited protective function on experimental hepatic fibrosis in rats,which may be attributed to the improving of fibrinolytic activity,eliminating of lipid peroxidation and enhancing of NO level.