1.Mesangial cells proliferation induced by high glucose is regulated by macrophage cytokine resistin via activating p38MAPK signal pathway
Jiao MU ; Defu JIA ; Fahuan YUAN
Chinese Journal of Nephrology 2008;24(11):832-837
Objective To investigate the effects of resistin on mesangial cells proliferation induced by high glucose and subsequent change of p38MAPK signal pathway. Methods Human macrophrages were cultured and treated with adenovirus encoding for resistin (Ad-resistin) for 48 h and were then co-cultured with human mesangial cells stimulated by high glucose for another 48 h. Mesangial ceils were harvested and their proliferation was measured by 3H-TdR. Activator protein 1 (AP-1) was examined by immunocytochemistry and laminin of excellular matrix was observed with immuofluorescence. Protein levels of p38MAPK and TGF-β1 were measured by Western blot. Smad2 phosphatase activity was aslo detected by Western blot. Results The mRNA and protein levels of resistin were significantly higher in Ad-resistin treated macrophages than those in Ad treated cells (P<0.01). Over-expression of resistin up-regulated p38MAPK protein levels of human mesangial cells(P<0.05). Resistin also promoted the proliferation of mesangial cells (P<0.01) and the synthesis of laminin stimulated by high glucose. The expression of TGF-β1 and phosphorylation of Smad2 were up-regulated in the mesangial cells (P< 0.05). Conclusion Macrophage cytokine resistin may promote mesangial cells proliferation and abnormal accumulation of excellular matrix stimulated by high glucose via activating p38MAPK signal passway.
2.Obtainment and Characterization of The Evolved Enzymes From Arabidopsis thaliana Glutathione S-Transferase Zeta Class
Sudan TAO ; Xiwen CHEN ; Jia LIU ; Xiangdong JIA ; Defu CHEN
Progress in Biochemistry and Biophysics 2006;0(02):-
The Arabidopsis thaliana glutathione S-transferases zeta class (AtGSTZ) is a multi-functional enzyme, which plays important role in cellular metabolism and environmental purification. Error-prone PCR and cycles of DNA shuffling were used to construct a mutagenesis library of AtGSTZ. The screening of the resultant libraries was carried out by a pH indicator dye-based colorimetric assay. Nine mutants which enhanced the dichloroacetic acid dechlorination activity were obtained. Among them, NN23 contained 25 amino acid substitutions with the activity improving 120%, whereas NN20 contained 24 amino acid substitutions with the activity improving 102%. EC1 contained 2 amino acid substitutions with the activity improving 47%. The rest 6 mutants contained one amino acid substitution with their activity increasing from 9% to 60%. The enzymatic characterization showed that all the evolved enzymes increased their catalytic efficiencies towards dichloroacetic acid and binding affinity towards glutathione whereas some of them increased the renaturability. However there is no obvious change in their thermostability. Based on these data, functional residues related to catalysis and refolding of AtGSTZ were discussed.
3.Application of ultrasound-guided transversus abdominis plane block for operations of inguinal region in infants
Yongsheng QIU ; Defu ZHANG ; Yingping JIA ; Xing LI ; Qing XU
Chinese Journal of General Practitioners 2014;13(12):1016-1018
One hundred and fifty ASA Ⅰ-Ⅱ infants scheduled to undergo surgery on unilateral inguinal region,were randomly assigned to 3 groups with 50 in each group.In groups Ⅰ and Ⅱ puncture was performed at traversus abdominis plane under ultrasound-guidance,0.4 or 0.5/kg of 0.25% ropivacaine was injected respectively.In group Ⅲ the triangle of Petit was positioned through palpating and then 0.5 ml/kg of 0.25% ropivacaine was injected.HR,RR,SpO2,PETCO2,expired sevoflurane concentration and BIS value at the time of entering the operation room (T1),incision of skin (T2),pulling hernia sac (T3),ending the surgery (T4),waking (T5),the number of pressing the analgesia pump after operation were recorded.The results showed that HR at T2 and T3 of groups Ⅰ and Ⅱ was lower than group Ⅲ (P < 0.05).The were significant differences in case numbers of insufficient intraoperative analgesia among 3 groups(x2 =10.500,P =0.005).The CHEOPS scores and the number of pressing analgesia pump after operation in groups Ⅰ and Ⅱ were lower than those in group Ⅲ (x2 =7.230,P =0.027).Results indicate that ultrasound-guided transversus abdominis plane block is safer and more effective than conventional method for operations of inguinal region in infants; it may reduce dose of local anesthetics and postoperative use of analgesics.