1.Potassium sodium dehydroandroan drographolide succinate combined with routine therapy for rotavirusenteritis in children
Wenliang ZHANG ; Xuemei RAO ; Lanfang LUO
International Journal of Traditional Chinese Medicine 2016;38(9):814-816
Objective To evaluate the efficacy of potassium sodium dehydroandroan drographolide succinate (PSDS) combined with routine therapy for rotavirus enteritis in children.MethodsA total of 148 children with rotavirus enteritis were included and divided into an observation group and a control group by random number table method, 74 in each group. The children in the observation group were treated with intravenous PSDS combined with routine therapy, and those in the control group with intravenous ribavirin combined with routine therapy. Serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay, and plasma lactate dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB (CK-MB) were determined using standard clinical laboratory procedures. The clinical efficacy was evaluated. Results The total efficacy rate in the observation group was significantly higher than that in the control group (91.9%vs. 78.4%;χ2=2.314,P<0.05). After the treatment, the serum levels of IL-6 (18.24 ± 3.62 ng/mlvs. 25.36 ± 5.25 ng/ml; t=2.425,P<0.05) and TNF-α (20.86 ± 4.28 ng/mlvs. 31.22 ± 7.15 ng/ml;t=2.503,P<0.05), and the plasma levels of LDH (104.25 ± 22.06 U/Lvs. 150.26 ± 37.22 U/L;t=2.316,P<0.05), CK (84.25 ± 13.57 U/Lvs. 107.88 ± 16.28 U/L;t=2.327,P<0.05) and CK-MB (22.30 ± 4.24 U/Lvs. 32.26 ± 7.14 U/L;t=2.426,P<0.05) in the observation group was significantly lower than those in the control group. The time to diarrhea resolution (2.42 ± 0.53 dvs.3.56 ± 0.78 d;t=2.316,P<0.05) and the time to fever resolution(2.11 ± 0.32 dvs.2.63 ± 0.43 d;t=2.472,P<0.05) in the observation group were significantly delayed than those in the control group, and the hospital length of stay longer (6.23 ± 1.42 dvs. 4.35 ± 0.96 d;t=2.413,P<0.05).Conclusions PSDS combined with routine therapy may reduce inflammatory response, protect from myocardial injury, and promote recovery in children with RVE.
2.Characters of location in cells and binding activity of chicken Ii and B-L gene ex-pressed products
Lanfang LUO ; Weiyi YU ; Fangfang CHEN
Chinese Journal of Immunology 2015;(7):879-883,889
Objective:To study characters of the location in cells and binding activity of chicken Ii and B-L gene expressed products.Methods:The cloned gene segments of chicken Ii,B-LA and B-LB were respectively inserted into prokaryotic or eukaryotic expression plasmids,and then these recombinant plasmids were respectively alone transfected or cotransfected into engineering bacteria, Rosetta(DE3) or 293T cells.All of the recombinant bacteria were induced to express and their products then were renatured.The singly expressed products were detected to their immunogenicity with Western blot, and the co-expressed products were tested their binding with pull-down method and ( SDS-) PAGE.Results:First six of prokaryotic and eukaryotic recombinant expression plasmids were con-structed.The eukaryotic expressed products of Ii, B-LA and B-LB genes located in cellular plasma membrane.The single protein molecules were achieved from prokaryotic expressed products, which were renatured and purified with a Ni-column respectively.Secondly the prokaryotic expression Ii,B-LA and B-LB molecules could respectively induce mouse to product specific anti-bodies, which could recognize the corresponding products in eukaryotic expression.This indicated that they retained their antigenicity.Finally with pull-down from the products in co-expression in engineering bacteria and renaturation the Ii/B-LA or Ii/B-LB complexes were purified and the Ii,B-LA or B-LB monomers were dissociated from these complexes after a SDS treatment.Conclusion:The prokaryotic or eukaryotic expressed products of chicken Ii and B-L genes could retain their antigenicity,and chicken Ii could bind B-L molecules after prokaryotic coexpression and renaturation.These results provide a useful method to study the relation between Ii and MHC molecules.
3.Research of functional segments of chicken B-FA molecule binding with Ii chain
Dandan YU ; Qiong WU ; Lanfang LUO ; Weiyi YU ; Fangfang CHEN
Chinese Journal of Immunology 2016;32(10):1413-1418
Objective: To research the functional segments of B-FA molecule binding invariant chain and their characters. Methods:The DNA segments (α1α2, sα1α2 and α3TC ) of B-FA genes were respectively cloned and inserted into prokaryotic or eukaryotic expression plasmids,then they were singly or co-transfected with Ii gene into the engineering bacteria E. coli (BL-21)or 293T cells. After induction of expression,affinity chromatography and SDS-PAGE identification,the binding between B-FA segments and Ii molecule and co-localization in cells were observed with Pull-down and Western blot. Results:First three recombinant prokaryotic expression plasmids and four recombinant eukaryotic expression plasmids were constructed. The single molecules expressed by B-FA segments were observed after an affinity chromatography. Secondly the complexes of Ii/B-FA-α1α2 and Ii/B-FA-sα1α2 were detected by a Pull-down from the co-transfected corresponding prokaryotic expression plasmids,but no complex of Ii andα3TC,also in the western blot it was detected that B-FA-α1α2 or B-FA-sα1α2 as functional segment could bind Ii to form complex. Finally in eukaryotic expression 293T cells B-FA-sα1α2 kept localization, the same as B-FA. Conclusion: Chicken B-FA-α1α2 is function segment to bind with Ii molecule and keeps the location characters same as B-FA. The results of this research first time provide experimental evidence about B-FA functional region binding segment to Ii molecule.
4.Correlation between Toll-like receptor 4 single nucleotide polymorphism and sepsis in Chinese patients of Han nationality
Jing JIAO ; Lanfang ZHU ; Zhe LUO ; Changhong MIAO
Chinese Journal of Anesthesiology 2014;34(z1):62-66
Objective To investigate the correlation between Toll-like receptor 4 (TLR4) single nucleotide polymorphism (SNP) and the risk,severity and prognosis of sepsis in Chinese patients of Han nationality.Methods One hundred and three Han nationality patients who developed sepsis after surgery,aged 18-80 years,were enrolled in the sepsis group,and 114 Han nationality patients without sepsis after surgery,aged 18-80 years,were enrolled in the control group.Venous blood samples were taken from the peripheral vein and three SNPs in TLR4 gene,rs10759932,rs11536889 and rs2737190,were genotyped by matrix-assisted laser desorption ionization time of flight mass spectrometry analysis.Correction for Logistic regression analysis was made to eliminate the effects of sex,age,underlying diseases and operation methods.The correlation between genotypes of SNP and occurrence of sepsis,organ dysfunction,septic shock and death from sepsis was analyzed.The odds ratio (OR) and 95% confidence interval (Cl) were calculated.Results Compared with the control group,there was a significant difference in genotype frequency ratios of rs10759932 (P < 0.05),but there was no significant difference in genotype frequency ratios of the other two SNPs in sepsis group (P > 0.05).There was correlation between rs10759932 and the occurrence of sepsis,and the variant allele (CT + CC genotypes) of rs10759932 increased the risk of sepsis (OR =1.86,95% Cl 1.17-2.97,P < 0.05).There was no correlation between the three SNPs and sepsis-related organ dysfunction,septic shock and death from sepsis (P > 0.05).Conclusion There is correlation between the variant allele of TLR4 rs10759932 and the increase in risk of sepsis after surgery in Chinese patients of Han nationality.
5.Effect of aminooxoacetic acid on focal cerebral ischemia injury in rats
Guofeng LI ; Jianxin ZHANG ; Haikun LUO ; Lanfang LI ; Qinzeng ZHANG ; Lijun XIE ; Liping LI ; Na HAO ; Weiwei WANG
Chinese Journal of Anesthesiology 2011;31(8):984-986
ObjectiveTo evaluate the effect of aminooxyacetic acid on focal cerebral ischemia injury in rats.MethodsEighty healthy male SD rats aged 2.5 month weighing 250-280 g were randomly divided into five groups( n = 16 each):sham operation group(group S),cerebral ischemia group(group Ⅰ),aminooxoacetic acid low,medium and high dose groups(groups AL,AM and AH ).Focal cerebral ischemia was induced by occlusion of middle cerebral artery using a nylon thread with rounding tip which was inserted into right internal carotid artery in groups I,AL,AM and AH.Intraperitoneal amincoxoacetic acid 25,50 and 100 μmol/kg were administered at 3 h of ischemia in groups AL,AM and AH respectively,while equal volume of normal saline 1 ml/kg were injected in groups S and I.Neurological function was assessed and scored (0= no deficit,4= unable to move,unconscious) in 8 rats at 21 h after aminooxyacetic acid administration in each group.The animals were then sacrificed and the brains were removed for determination of the cystathionine beta-synthase (CBS) activities in cortex,hippocampus and striatum corpora.The other eight rats of each group were sacrificed at 21 h after amincoxoacetic acid administration for determination of the cerebral infarct volume.ResultsCompared with group S,the neurological deficit scores and the CBS activities in cortex and hippocampus were significantly increased,the infarct volumes were significantly enlarged in group Ⅰ ( P < 0.05).Compared with group Ⅰ,the neurological deficit scores and the CBS activities in cortex and hippocampus were significantly decreased,the cerebral infarct volumes were significantly reduced in groups AM and AH (P < 0.05).There was no significant difference in the above-mentioned variables between groups AL and Ⅰ.ConclusionAminooxoacetic acid can reduce focal cerebral ischemia injury by decreasing CBS activity and reducing H2 S production in rats.