5.Construction and Functional Analysis of a Synthetic Wound-and Hormone-inducible Promoter ocs/mas
China Biotechnology 2006;0(07):-
The use of optimal regulatory sequences for simultaneous expression of the transgenes might play a significant role in engineering plants with increased disease and insect resistance.The plant expression vector pOMS-GUS,which contained the GUS gene under the control of a chimeric promoter based upon the mannopine synthase(mas)promoter and the octopine synthase(ocs)enhancer,was constructed.Used as control,another vector pMAS-GUS,carried the GUS gene driven by only the mas promoter.The two vectors were introduced into tobacco plants by Agrobacterium-mediated transformation.Fluorometric assays for GUS activity and reverse transcription-polymerase chain reaction(RT-PCR)analysis revealed that GUS gene expressed weakly with untreated transgenic tobacco while the level of GUS activity increased steadily after 1 h subjected to wounding.The expression of the mas and ocs/mas promoters was induced a further 1.8-fold and 5.7-fold,respectively.SA(1 mmol/L)or MJ(250 ?mol/L)treatment also caused a large induction of the ocs/mas chimeric promoter;And the application of SA in combination with MJ(1 mmol/LSA & 250 ?mol/L MJ)produced an additive effect that exceeded the wounding response.The results showed that the ocs/mas chimeric promoter is a strong inducible promoter that can be activated by various stresses.The chimeric promoter should have utility in development of disease and insect resistant transgenic crops.
7.Cystic fibrosis in 2 children with Pseudomonas aeruginosa infection
Mengni LI ; Huan LEI ; Juan FAN ; Min LI ; Lan LI
Chinese Journal of Applied Clinical Pediatrics 2021;36(4):300-302
Clinical data of 2 children with cystic fibrosis (CF) and Pseudomonas aeruginosa infection in the Department of Pediatrics of Sichuan Provincial People′s Hospital from April 2018 to June 2019 were retrospectively analyzed.Patient 1 was an 11-year-old girl with no history of recurrent respiratory infections. Pseudomonas aeruginosa was found in the first sputum culture.A large number of yellow and white secretions were visible under repeated fiberoptic bronchoscopy.Chest CT showed multiple spots and tree-in-bud signs around the bronchi of both lungs. CFTR gene test results revealed 3 heterozygous mutations: c.2909G>A (chr7: 117246728), c.*133T>A (chr7: 117307295) and c. *125delT (chr7: 117307285). The other patient was a 7-year-old boy with a history of recurrent respiratory infections.His parents were close relatives.Multiple cultures of sputum and bronchoalveolar lavage fluid of the boy were Pseudomonas aeruginosa, and chest CT suggested dilation and inflammation in bronchi of both lungs.Gene detection showed that the c. 380T>G homozygous mutation at chromosome chr7-117171059 resulted in an amino acid change p. leu127stop (nonsense mutation). This article suggests that CF should be considered for Pseudomonas aeruginosa infected children having signs of bronchiectasis on chest CT and a large number of secretions under bronchoscopy.Besides, it is necessary for such kind of children to perform genetic testing in time to confirm the diagnosis as soon as possible.
8.Ultrasonic cardiac output monitor for evaluation of cardiac function in neonatal sepsis
Juan FAN ; Jing LIAO ; Mengni LI ; Lan LI
Chinese Pediatric Emergency Medicine 2017;24(8):599-602
Objective To monitor the change of cardiac function in neonatal sepsis by using the ultrasonic cardiac output monitor(USCOM).Methods Thirty two cases of mild sepsis neonates and nineteen cases of severe sepsis neonates were enrolled and thirty three cases of healthy neonates were enrolled in the control group.The cardiac output indicators of three groups were monitored by USCOM.The differences of cardiac function among 3 groups and the changes of candiac function after treatment in septic neonates were investigated.Results Compared with control group,the heart rate,systemic vascular resistance of mild sepsis neonates group and severe sepsis neonates group were significantly higher,and the cardiac output,systolic volume,cardiac index,aortic peak velocity were significantly lower,the differences were statistically significant(P<0.05).The cardiac index,cardiac output were significactly lower in severe sepsis neonates group than those in mild sepsis neonates group(P<0.05).After improving cardiac function treatment,all cardiac function indexes of sepsis neonates were improved than before treatment,the differences were statistically significant(P<0.05).Conclusion There are significant cardiac function changes in sepsis neonates.The cardiac function and overall circulation state monitoring by USCOM is fast and convenient,and USCOM can provide the basis for treatment and evaluation of the neonatal sepsis.
9.Survey on the genes conferring plasmid-mediated quinolone resistance and those encoding β-lactamases in Escherichia coli isolates
Fangjun LAN ; Juan WU ; Qingwen HE ; Yingping CAO ; Bin LI
Chinese Journal of Infection and Chemotherapy 2017;17(3):293-297
Objective To examine the prevalence of plasmid mediated quinolone resistance (PMQR) genes and their correlation with the genes encoding β-lactamases in E.coli isolates.Methods A total of 200 levofloxacin-and/or ciprofloxacin-resistant E.coli isolates were collected from Fujian Medical University Union Hospital during the period from July to December 2013.PCR method was used to screen these E.coli isolates for the presence of qnrA,qnrB,qnrC,qnrD,qnrS,aac(6')-Ib-cr,qepA,oqxAB genes,and the blaTEM,blasnv and blacTx-M genes in the PMQR positive strains.Agar dilution method was utilized to measure the antimicrobial susceptibility of PMQR-positive strains.Phylogenetic analysis was conducted by triplex PCR.Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) was used to evaluate the genetic similarity between the PMQR-positive isolates.Results Of the 200 clinical isolates of E.coli,58 (29.0%)were PMQR-positive.And qnr,aac(6')-Ib-cr,oqxAB,and qepA genes were positive in 11 (5.5%),41 (20.5%),16 (8.0%),1 (0.5%) strains,respectively.The genes encoding CTX-M-1,CTX-M-9 and TEM type enzymes was positive in 32 (55.2%),17 (29.3%),and 1 (1.7%) of the PMQR-positive strains,respectively.The blasHv gene was not identified in any isolate.PMQR-positive strains were multi-drug resistant.Phylogenetic analysis indicated that 21 (36.2%),17 (29.3%),11 (19.0%),and 9 (15.5%) of the PMQR-positive strains belonged to group A,group D,group B2 and group B 1,respectively.ERIC-PCR suggested the PMQR-positive isolates belonged to 50 different types.Only one strain was non-typeable.Conclusions Most of the PMQR-related genes in E.coli are aac(6')-Ib-cr,qnr,and oqxAB in our hospital,which are highly relevant to β-1actamase genes.PMQR-positive strains may spread by way of non-clonal dissemination in our hospital.
10.Quantification of tryptase and TIM-3 double positive mast cells in human chronic periodontitis tissue
Juan LI ; Hua JIN ; Tian LAN ; Shiguang HUANG
Journal of Practical Stomatology 2014;(4):482-486
Objective:To quantify the tryptase and T cell immunoglobulin mucin domain-3(TIM-3)double positive mast cells in hu-man chronic periodontitis tissue using double immunofluorescence staining.Methods:25 healthy controls,28 chronic mild periodontitis and 30 chronic advanced periodontitis patients were included.The gingival specimens were stained with HE for histology,and with double immunofluorescence staining for the identification of tryptase and TIM-3 double positive mast cells in gingival tissue.Results:In chronic periodontitis tissue the degree of gingival inflammation was significantly increased,the densities(cells/mm2 )of tryptase and TIM-3 double positive mast cells were significantly increased(P<0.05),in addition,that in chronic advanced periodontitis group was significantly higher than in the chronic mild periodontitis group(P<0.05).Conclusion:Tryptase and TIM-3 double positive mast cells has the similar tendency as the severity of periodontitis inflammation in human periodontitis tissue.Tryptase and TIM-3 double positive mast cells may play an important role in human chronic periodontitis.