Résumé
Objective To investigate the effects of ulinastatin on renal expression of endothelin￣1 and nitric oxide (NO) in rats with toxic acute kidney injury(AKI). Methods Twenty￣four male SD rats were randomly divided into the normal control group,model control group and treatment groups, with 8 rats in each group. Except normal control group, rats were subcutaneously injected with gentamicin (300 mg.kg-1 .d-1 ) for 3 days to establish a model of toxic AKI.Rats in the treatment group were intraperitoneally injected with a 7￣day course of ulinastatin (30 000 U.kg-1 .d-1 ) from the fourth day.While the other two groups were injected with 0.9% sodium chloride injection (3 mL.kg-1 .d-1 ). The serum level of creatinine and cystatin￣C,urinary concentration of kidney injury molecule￣1(Kim￣1) and neutrophil gelatinase￣associated lipocalin (NGAL), level of endothelin￣1 and NO,expression of endothelin￣1 mRNA,endothelial nitric oxide synthase (eNOS),induced nitric oxide synthase (iNOS),eNOS mRNA and iNOS mRNA in homogenate of renal tissues in each group were detected on the eleventh day. Results Compared with the normal control group,serum level of creatinine and Cystatin￣C,urinary concentration of Kim￣1 and NGAL,level of endothelin￣1 and NO,expression of endothelin￣1 mRNA,iNOS and iNOS mRNA in homogenate of renal tissues were higher in model control group (P<0.01, respectively), while which were lower in treatment group than those in model control group ( P < 0. 01, respectively). And no statistical significant difference of eNOS and eNOS mRNA expression in homogenate of renal tissues existed among the three groups. Conclusion Ulinastatin possesses curative role against rat with toxic AKI via down￣regulating renal expression of endothelin￣1,NO and iNOS.
Résumé
Objective To explore the curative effect of ulinastatin against toxic acute kidney injury(AKI) in rats and its mecha-nism .Methods Twenty-four male SD(Sprague Dawley) rats were randomly divided into 3 groups ,control group ,model group and treatment group with 8 rats in each group .Rats were subcutaneously injected gentamicin(300 mg/kg of body weight per day) for 3 days to establish models of toxic AKI .Rats in treatment group were intraperitoneally injected with a 7-day course of ulinastatin(30 000 U/kg of body weight per day) from 4th day .Dectetion of serum level of creatinine and Cystatin-C(Cys C) ,urinary concentra-tion of kidney injury molecule-1 (Kim-1 ) and neutrophil gelatinase-associated lipocalin (NGAL ) ,activity of superoxide dismutase (SOD) and glutathione peroxidase(GSH-Px) ,content of malondialdehyde ,levels of tumour necrosis factor-alpha(TNF-α) and inter-leukin-1β(IL-1β) in homogenate of renal tissues as well as observation of renal pathological changes and semiquantitative score in each group were conducted on 11th day .Results In model group ,degeneration and necrosis of renal tubular epithelial cell ,dilatation of renal tubular cavity and inflammatory cell infiltration in renal interstitial were observed .Renal pathological changes were milder in treatment group ,when compared with the model group .Renal pathological semiquantitative score ,serum level of creatinine and Cys C ,urinary concentration of Kim-1 and NGAL ,content of malondialdehyde ,levels of TNF-α and IL-1β in homogenate of renal tissues were higher in model group than in control group ,while those in treatment group were lower than in model group(P<0 .01 , respectively) .And activity of SOD and GSH-Px in homogenate of renal tissues were lower in model group than in control group ,and those in treatment group were higher than in model group and control group(P<0 .01 ,respectively) .Conclusion Ulinastatin pos-sesses a curative role in toxic AKI in rat via inhibiting oxidative stress and down-regulating levels of proinflammatory factor in renal tissues .
Résumé
Objective To construct the recombinant protein AtlM ( rAtlM) of Staphylococcal au-reus through prokaryotic expression system and to investigate its antibacterial activity in vitro.Methods The specific primers were designed according to atlM gene sequence of Staphylococcal aureus recorded in Gen-Bank, and atlM gene was amplified by PCR from the Staphylococcal aureus strain (ATCC25923).The re-combinant plasmid pET-32а(+)/atlM was constructed and transformed into Transetta ( DE3 ) to express AtlM after induced by IPTG .The expressed protein AtlM was further analyzed by SDS-PAGE and purified by electroeluting of bag filter.The minimal inhibitory concentrations (MICs) of rAtlM to ATCC25923 and oxac-illin-resistant S.aureus strain were determined by the broth microdilution method .S.aureus ATCC25923 strain and oxacillin-resistant S.aureus strain (final concentration of 5×105 CFU/ml) were exposed to rAtlM (50 μg/ml) respectively to test its antibacterial activity in vitro.Results The recombinant protein AtlM was expressed and purified successfully with a relative molecular weight of 80 ×103 and a concentration of 1.25 mg/L.The MICs of rAtlM to ATCC25923 strain and oxacillin-resistant S.aureus strain were 8 μg/ml and 64 μg/ml, respectively.In vitro test showed that rAtlM had inhibitory effects on the growth of ATCC25923 strain and oxacillin-resistant S.aureus strain after 1 h of intervention (P=0.004 and P=0.026, respectively), which lasted to 5 h for ATCC25923 strain (P=0.012) and 3 h for oxacillin-resistant strain (P=0.001).Conclusion This study shows that rAtlM has a certain antibacterial effects on S.aureus ATCC25923 strain and oxacillin-resistant S.aureus strain, suggesting a possibility of serving as antimicrobial agent.
Résumé
Objective To obtain the pneumococcal autolysin(LytA)and choline binding protein A(CbpA)by prokaryotic expression system and investigate their diagnosis for infection caused by Streptococcus pneumoniae.Methods The specific primers were designed according to lytA and cbpA of Streptococcus pneumoniae gene sequence.lytA and cbpA were amplified by PCR form the pneumococcus genome.After IPTG inducing,the recombinant proteins were purified by electroeluting of bag filter,detected by SDS-PAGE and Western blot.Serum lgG and IgM antibodies accordingly of BALB/c mice infected with Streptococcus pneurnoniae were detected by ELISA.Results The recombinant plasmid pET-32a(+)/lytA and pET-32a (+)/cbpA were constructed successfully.Fusion proteins LytA and CbpA were expressed and displayed expected antigenicity.IgM and IgG antibodies level anti LytA were significantly higher than the control group (infections with B Streptococcus group and healthy mice),(P0.05).Diagnostic sensitivity of CbpA was 83.3%(IgG)and 75.0%(IgM).Diagnostic specificity of LytA was 100%(IgG and IgM).Conclusion The synergistic use of specificity of LytA and sensitivity of CbpA may be worthy of serological diagnosis for Streptococcus pneumoniae infection,and may be used for further clinical test.
Résumé
Objective To explore the value of combined measurement of urine N-acetyl-beta-D-glucosaminidase (NAG) activity and serum Cystatin C in diagnosing diabetic nephropathy (DN) in early phase. Methods Sixty-two cases with type 2 diabetes (diabetic group) were divided into three groups according to their 24 hours urinary albumin excretion (24hUAE) : group A (normal albuminuria, 20 cases), group B (microalbuminuria, 22 cases) and group C (macroalbuminuria, 20 cases). Furthermore, 30 healthy people were involved in control group. 24hUAE,NAG,serum creatinine (SCr) and serum Cystatin C were measured, and endogenous creatinine clearance rate (Ccr) was calculated by Cockcroft-Gault formula. All these indexes among three groups were compared. Results The levels of urinary NAG activity and serum Cystafin C in diabetic group was significantly higher and Ccr was significantly lower than those in control group(P < 0.01). The levels of urinary NAG activity and serum cystatin C gradually increased in group A, B and C(P< 0.05 or < 0.01). While no significant difference was observed between group A and group B in the level of SCr (P > 0.05). There were significant positive correlations among the levels of urinary NAG activity, serum Cystatin C,24hUAE and SCr (P< 0.01),and all above showed negative correlations with Ccr (P<0.01). Co-detection of urinary NAG activity and serum Cystatin C had significantly higher positive rate [80.6%(50/62)] than single one [58.1%(36/62),61.3%(38/62)](P<0.05). Conclusion Co-detection of urinary NAG activity and serum Cystatin C may indicate early renal damage in DN, and it is valuable in diagnosing DN in early phase.
Résumé
sing guidewire through temporary catheter is an alternative strategy, because of avoiding this avoids vein re-puncturation and doesn't increase catheter related complication.