1.Ghrelin attenuates renal fibrosis and apoptosis of obstructive nephropathy
Guangxi SUN ; Lipei FAN ; Xianghua SHI ; Min LI ; Ming ZHAO
The Journal of Practical Medicine 2015;(7):1102-1106
Objective To investigate the effect and underlying mechanisms of ghrelin in a rat model of renal fibrosis. Methods Male Sprague-Dawley rats were divided into 4 groups , including sham operation +saline or brain gut peptide treatment group , model + saline or brain gut peptide treatment group. Unilateral ureteral obstruction (UUO) was established by left ureteral ligation. 7 days and 14 days after operation, the rats were sacrificed , while the kidney tissue of obstruction side was harvested for pathlogical changes through Masson coloration. Expression of α-smooth muscle actin (α-SMA), transforming growth factor beta1 (TGF-β1) and phosphorylated Smad3 (p-Smad3) in renal tissues were analyzed through immunohistochemistry. Expression of α-SMA and TGF-β1 mRNA was detected by real-time-PCR. Apoptosis kidneys cells were marked with TUNEL. Results Ghrelin inhibited renal fibrosis by reducing the production of collagen , restraining extracellular matrix (ECM) deposition and decreasing the expression of α-SMA. Meanwhile, ghrelin inhibited the accumulation of myofibroblasts by blocking the transforming growth factor-β1/Smad3 (TGF-β1/Smad3) signaling pathway. Moreover, ghrelin could attenuate renal tubular cell apoptosis induced by UUO injury. Conclusion Ghrelin can reduce renal fibrosis and renal cell apoptosis induced by UUO , demonstrating that ghrelin is a potent antifibrotic agent that may have therapeutic potential for patients with obstructive nephropathy.
2.Effect of NGF on the sperm motility of human in vitro
Kai LIN ; Cuige SHI ; Yongqi ZHAO ; Shuhong LIU ; Ming FAN
International Journal of Laboratory Medicine 2012;33(20):2433-2437
Objective Motility is an important physiological characteristic of a mature sperm.Nerve growth factor(NGF) is a protein essential for the development,maintenance and survival of the peripheral and central nervous systems.NGF and its receptors TrkA and p75 are widely expressed in the testis,accessory reproductive organ,and the epididymal sperms.In the present study,we investigated the role of NGF on human sperm motility.Methods Use 0.1,1 and 10 μmol/L concentrations of NGF,on sperm motility study to investigate the optimal concentration.Use CASA to detect Sperm motility changes every 10 minutes in an hour after 10 μmol/L NGF was added to the semen.Results The parameters of sperm motility increased after NGF incubation had significant difference, in particular,VAP,VSL,VCL,BCF and LIN mean were significantly increased more than 32%.MAD,STR,ALH and WOB mean had no notable difference.Furthermore,NGF promotes the sperm motility in a time- and dose- dependent manner.In addition,the enhancement of NGF on sperm motility was more stronger than those of sperm culture medium.Conclusion Our findings suggest that NGF plays a promoted role in human sperm motility.
3.Effects of irradiation of intense pulsed light on the collagen metabolism in human fibroblasts
jin, MA ; shi-ming, GAO ; zhi-hong, FAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To investigate the effects of intense pulsed light(IPL) on the collagen metabolism in human fibroblasts.Methods The callan foreskin fibroblasts were cultured and treated with different wave length and designed dose of IPL irradiation(590-1 200 nm,and 29 J/cm2),while 10 without IPL irradiation as controls.By culturing 1,12,24 and 48 h after the irradiation,the synthesis of collagen was measured by 3H-proline incorporation determination. Results The synthesis of collagen activity increased significantly(P
5.Study of Bupropion Hydrochloride Tablet and Fluoxetine in Treatment of Depression Multicenter Clinical Trial
hua-fang, LI ; shi-ping, XIE ; ming, LI ; jian-an, SHI ; xiao-ling, SHEN ; jian-xiong, FAN ; niu-fan, GU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
0.05), CGI and HAMA. Safety analysis suggested that no significant differences were found in symptoms and frequency of side effects between the two groups.Conclusion Bupropion hydrochloride tablet is an effective and safe antidepressant. It has similar effect and safety compared with fluoxetine in the treatment of depression.
7.Interaction of anti-thrombotic and anti-inflammatory activities of commonly used traditional Chinese medicine for promoting blood circulation and removing blood stasis revealed by network pharmacology analysis.
Ming Lü ; Taiyi WANG ; Xiaoxuan TIAN ; Xinhui SHI ; Guanwei FAN ; Yan ZHANG ; Yan ZHU
Acta Pharmaceutica Sinica 2015;50(9):1135-41
Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.
8.Construction and expression of anti-CD3?anti-CD19 bispecific diabody and analyzing its specific binding activity
Wei LI ; Dongmei FAN ; Xin CHENG ; Ruizan SHI ; Rong LIU ; Simei REN ; Min WANG ; Ming YANG
China Oncology 1998;0(04):-
Background and purpose:Bispecific antibody is a special pattern of genetic engineering antibody.It possesses 2 antigen binding sites, one directed at the effecter cells and the other at the target cells which are always the tumor cells.This enables the effecter cells to recruit to the targets efficiently and exert cytotoxic effect.The experiment was aimed to construct the expression vector of anti-CD3?anti-CD19 bispecific antibody, purification and analyzing the activity of the diabody.Methods:Applyed PCR and overlap PCR to construct the expression vector of anti-CD3?anti-CD19 bispecific antibodies, transformed it into E.coli 16C9 for expression.Purified the diabody by His-tag affinity chromatography and confirmed by 12% SDS-PAGE and Western blot.Antigen binding activity of the diabody was analyzed by FACS.Results:The DNA sequence of the expression vector containing anti-CD3?anti-CD19 fragment was comfirmed.The yield of purified diabody was up to 5 mg/L.The molecular weight of diabody was correctly indicated by SDS-PAGE and Western blot.The diabody could specifically bind to CD19+ Raji cells and CD3+ Jurkat cells while competitively inhibit binding of HIT3a and HIT19a to the cells.Conclusion:Successfully constructed the expression vector of anti-CD3?anti-CD19 bispecific antibodies, the diabody acquired by the method could be produced with high level expression and also had high specific affinity with CD19+ Raji cells and CD3+ Jurkat cells.
9.Development of excitotoxicity model with primary hippocampal neurons and implications of viability essay
Jie SHEN ; Changkai SUN ; Ming FAN ; Hui MA ; Ping LIN ; Yuying TONG ; Guangxia SHI
Chinese Journal of Laboratory Medicine 2003;0(10):-
Objective To study the injury and protective action of drugs on neurons, the model of glutamate excitotoxicity on primary cultured hippocampual neurons from new born rats were( set up. Methods)By use of trypan blue dye staning and testing the lactate dehydrogenase leakage from cultured neurons, to investigate the neuron survival rate. Results We found the injury of neurons was related with the concentration of glutamate. NMDAR non-competitive antagonist —MK-801 could protect the glutamate excitotoxic damage on neurons. Conclusion The glutamate results in neuron injury through NMDAR; the model of neuron culture was sufficient for glutamate-induced excitotoxicity.
10.Detection and analysis of plasmid-mediated carbapenem-hydrolyzing enzyme KPC-2 in carbapenem-resistant Escherichia coli
Shougang KUAI ; Haifeng SHAO ; Weiping WANG ; Lining SHI ; Xiaowei ZHANG ; Ming FAN
Chinese Journal of Laboratory Medicine 2009;32(10):1120-1123
Objective To study molecular epidemiology and carbapenem-resistance mechanism of four Escherichia coli strains isolated from general surgery wards. Methods Antibiotic susceptibility was carried out by K-B gar diffusion and agar dilution methods. Carbapenemases were screened by three dimensional test and EDTA-Na_2-disk synergy test. Pulsed-field gel electropboresis (PFGE) was performed to analyze molecular epidemiology of isolates. Plasmid was extracted by using an alkalinelysis technique. Conjunction experiment, transformation assay, specific PCR and DNA sequencing were performed to confirm carbapenemase genotype and its transmission mechanism Results Four Escherichia coli isolates were resistant to most antimicrobials including carbapenem. PFGE showed that the four isolates belong to four different clonal strains. Specific PCR and DNA sequence analysis identified that carbapenem resistance in four clinical isolates was mediated by KPC-2 encoded on an approximately 56 000 bp plasmid, and this plasmid did not harbor aminoglycosides and fluorquinolones resistant genes. Conclusion Four Escherichia coli isolates with carbapenem resistance are obtained from our hospital, and KPC-2 plasmid is main cause of carbapenem resistance in these isolates.