6.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.Botulinum neurotoxin type A heavy chain intervenes in expression of ge-neral proteins in spinal cord after unilateral spinal cord injury in rats
Jing LAN ; Hong WANG ; Juan BAI ; Yafang WANG ; Xiaqing LI
Chinese Journal of Pathophysiology 2016;32(12):2125-2132
AIM:To observe the effect of botulinum neurotoxin type A heavy chain ( BoNT/A HC) on the pat-tern of spinal protein expression by intrathecal injection after spinal cord injury in rats , and to explore the role of BoNT/A HC intervention in spinal protein expression and some of its mechanisms in nerve regeneration after injury .METHODS:The model of unilateral lumbar spinal cord injury was established .The effects of BoNT/A HC intervention at different doses (2 μg, 4 μg, 6 μg and 8 μg) on the general pattern of protein expression in the spinal cord tissues at the injury site and the cranial part adjacent to the injury site was measured and evaluated by SDS-PAGE and Coomassie brilliant blue staining first, and then by two-dimensional SDS-PAGE.RESULTS:The histological structure of the ipsilateral side of lumbar spi-nal cord showed obvious destruction and degradation , mainly affecting both gray and white matter of the left side of the cord.The result of SDS-PAGE with Coomassie brilliant blue staining from injured spinal cord tissue displayed that the ex-pression of some proteins after one-time BoNT/A HC treatment appeared obviously different from that without BoNT /A HC treatment.Moreover, the pattern of the protein expression affected by BoNT/A HC was similar to that of the normal spinal cord.The more detail information from two-dimensional SDS-PAGE indicated that more than 10 proteins with different mo-lecular weight and isoelectronic points were differentially expressed at day 2 and day 20 after local injection of 6μg BoNT/A HC.This altered expression actually appeared a tendency toward the pattern shown in normal group .CONCLUSION:The immediate application of BoNT/A HC at the injury site after unilateral lumbar spinal cord injury is able to affect the pattern of local protein expression .The altered protein expression by injury could be reversed back to normal or approxi -mately normal by local BoNT/A HC administration.
10.Spatio-temporal expression of P2X3 receptor in rats with diabetic mechanical allodynia
Yuanyuan CUI ; Huanghui WU ; Lan WANG ; Juan SHI ; Yunqing LI
Acta Anatomica Sinica 2014;(4):540-544
Objective To investigate the spatio-temporal expression of P2X3 receptor ( P2X3R) in rats with diabetic mechanical allodynia ( DMA ) .Methods DMA model in rats was induced by intraperitoneal injection of streptozocin ( STZ) .The von Frey filaments were applied to identify the changes of the paw withdrawal threshold ( PWT) in DMA rats.Immunofluorescence was employed to detect the spatio-temporal expression of P2X3R in the spinal dorsal horn (SDH), dorsal root ganglion (DRG) and hind paw skin on different time points after intraperitoneal injection of STZ , respectively.The protein expression of P2X3R in SDH and DRG was further semi-quantitatively analyzed by Western blotting.Results Compared with control group , intraperitoneal injection STZ induced significant mechanical allodynia indicated by the reduced PWT from 7 days, and which reached the peak on 14d and maintained to 28days (P<0.05). The expression of P2X3R in DRG neurons was significantly increased on 14 days and 21 days (P<0.05), while that in SDH and skin was markedly increased on 21 days and 28 days, compared with control group (P<0.05).Conclusion With the progress of DMA, the expression of P2X3R was significantly increased in the SDH, DRG and skin, which was almost parallel with the mechanical allodynia , but the changes in SDH and skin were 1 week later than that in DRG .These results suggest that P2X3R may play a key role in the maintenance of the DMA .