1.Construction of a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum and expression in Escherichia coli BL21 (DE3)
Li, ZHANG ; Wen-gui, LI ; Jin-ping, XIANG
Chinese Journal of Endemiology 2013;32(6):608-611
Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double-digestion technique confirmed that Sj26GST antigen gene was successfully cloned into pGEX-1λT vector,the relative molecular mass of the expressed recombinant protein was approximately 52 × 103 by SDS-PAGE,and the amount of expressed protein was 20% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pGEX-Sj26GST is successfully constructed and highly expressed in E.coli and the expressed fusion protein shows specific antigenicity.
2.Self-Control Study on 51 Children with Epilepsy Treated by Levetiracetam as Add-on Therapy
qiong-xiang, ZHAI ; juan, GUI ; yu-xin, ZHANG
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To evaluate the long-term clinical efficacy and security of levetiracetam (Lev) as add-on therapy in patients with different types of epilepsies from an observational study.Methods Fifty-one patients were evaluated (14 female,37male,age range from 7months to 16 years,mean age 8.7 years) with different types of epilepsies ( 20 complex partial seizure,10 tonic-clonic seizure,1 tonic seizure,6 myoclonic epilepsy,2 Lennox-Gastaut syndrome,4 infantile spasms and 2 unspecified epileptic syndromes).The basis for comparison was defined as the seizure frequency in the 3 months prior to the commencement of treatment.Patients received Lev as add-on therapy.The initial dosage was 20 mg/(kg?d),and it was increased 10 mg/(kg?d) every 2 weeks.The maintenance dosage was 30-40 mg/(kg?d).Seizure frequency changes and adverse events were observed.Follow-up was conducted for a period of 6.8 months after treatment.SPSS 14.0 software was used to compare the difference between the seizure frequency before the Lev treatment and that after the Lev treatment.Results Thirteen (25.5%) out of the 51 patients reduced seizure frequency,16 (31.4%) patients had no reoccurrence;While another 9 (17.6%) patients seizure frequencied were reduced,8 patients' remained the same,and 5 patients' condition was got wor-sened.Six cases ceased treatment because of the worsening of the disease and the intolerance of Lev.The difference and after seizure frequency before in Lev treatment is statistically significant(P
3.Character of Clinical and Active Electroencephalogram in Children with Nocturnal Paroxysmal Epilepsy
juan, GUI ; qiong-xiang, ZHAI ; yu-xin, ZHANG
Journal of Applied Clinical Pediatrics 2004;0(11):-
0.05).The period when epileptiform abnormalities appear was obviously different(P
4.Analysis of Positron Emission Tomography and Video Electroencephalogram in Children with Nocturnal Frontal Lobe Epilepsy
yu-xin, ZHANG ; juan, GUI ; qiong-xiang, ZHAI
Journal of Applied Clinical Pediatrics 2006;0(24):-
0.05).Of essay group 19 children whose PET were normal or slight abnormal,8 children's VEEG had epileptifrom abnormalities only appear in lucid interval,8 children's VEEG had epileptifrom abnormalities appear in nocturnal sleep period,3 children's VEEG had epileptifrom abnormalities appear in lucid interval and nocturnal sleep period.Of essay group,7 children whose PET were serious abnormal,6 children's VEEG had epileptifrom abnormalities appear in lucid interval and nocturnal sleep period.The PET outcome was relate with the time of VEEG epileptic discharge(r=0.461 P
7.The inhibitory effect of decorin on proliferation of rabbit epithelial cell
Jian-nan, XIANG ; Gui-lan, ZHANG ; Hai-jiang, ZHANG ; Guo-hua, WANG ; Ming, HUO
Chinese Journal of Experimental Ophthalmology 2012;30(1):41-45
BackgroundResearches found that the posterior capsular opacification (PCO) after lensextraction is associated with the elevation of the transforming growth factor-β(TGF-β).To seek the drug for inhibitingproliferation of lens epithelial cells (LECs) is crucial in the treatment and prevention of PCO.ObjectiveThisstudy was to investigate the preventing effects of decorin on the proliferation of LECs.MethodsRabbit LECs wascultured and passaged.The LECs in growth phase were incubated in 96 well plate at the density of 8×106/L.Decorinwith the concentrations 0.1,1.0,10.0 mg/L was added into the medium for 24,48 and 72 hours respectively.0.1%DMSO was used at the same way as positive control group,and the regular cultured cells worked as blank controlgroup.The inhibitory rates of different concentrations of decorin on the growth of LECs were detected by MTT at 24,48and 72 hours after addition of decorin.The percentage of LECs in different cell cycles in various groups was assayedusing flow cytometry.TGF-β level in medium suspension was detected using ELISA.The expression of TGF-β mRNA in LECs was checked by RT-PCR,and α-SMA expression in LECs was determined using immunochemistry.Results ELISA assay showed a statistical difference in the TGF-β levels of different groups (F=39.24,P=0.03 ).The TGF-β levels in 1.0,10.0 mg/L decorin groups were significantly decreased in comparison with blank control group (P<0.01) and 0.1 mg/L decorin group (P<0.05 ).The inhibitory rates of decorin in the concentrations of ≥ 1.0 mg/L on the growth of LECs were higher than the blank control group,and those in various concentrations of decorin groups were considerably lower in 24 hours compared with 48 and 72 hours ( P<0.05 ) and so was the 48 hours compared with 72 hours (P<0.05 ).The percentages of LECs in G0/G1 phase were ascent in 0.1,1.0 and 10.0 mg/L decorin groups in comparison with G2/M and S phase (P<0.05).Immunochemistry revealed the weak expression of α-SMA in various decorin groups in comparison with control group. Conclusions Decorin can effectively inhibit LECs growth and induce LECs apoptosis in concentration- and time-dependent manner.It is suggested that decorin can be used in the prevention and treatment of after cataract.
8.Treatment of infantile subglottic hemangioma with oral propranolol.
Li-xing TANG ; Ya-mei ZHANG ; Gui-xiang WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(9):735-738
OBJECTIVETo observe the efficacy of oral Propranolol in the treatment of infantile subglottic hemangioma.
METHODSEleven children (6 females and 5 males) with a median age at onset of treatment being 4 months were included in this study. Propranolol was given after laryngoscopy and a CT scan with contrast of the trachea confirming the presence of a subglottic hemangioma. The starting dose of Propranolol was 0.5 mg/kg per day, given in 2 or 3 divided doses. Heart rate and blood glucose were monitored during the treatment. If no side effects occurred, the dose was increased to 1 mg/kg per day at the third day and to 2 mg/kg per day at the sixth day. Treatment was continued at home after 10 days of inpatient treatment and the children were reevaluated monthly.
RESULTSAfter 24 - 48 hours of treatment, all of the children had improvement in their airway obstruction which was confirmed by fibro-laryngoscopy. The diameter of the subglottic stenosis from the hemangioma decreased from 3.9 - 5.0 mm to 1.5 - 2.0 mm,and the color was also lighter than before. In 3 children with cutaneous hemangioma, there was also significant improvement in the cutaneous lesions after treatment, with the color becoming lighter. There were no significant ECG, blood pressure or blood biochemical changes during the treatment. Two of the children had hypoglycemia at the first dose, but improved after blood transfusion and changing their diet. In five children, the treatment was stopped after 6 to 11 months when the obstructive symptoms improved. None of the children in this group had any evidence of recurrence. In the 5 children who stopped treatment, the obstructing mass in the subglottis was less than 10% of the diameter.
CONCLUSIONOral propranolol is a safe and effective treatment for infantile subglottic hemangioma. It may be used as a first-line therapeutic modality.
Female ; Glottis ; Hemangioma ; drug therapy ; Humans ; Infant ; Laryngeal Neoplasms ; drug therapy ; Male ; Propranolol ; therapeutic use ; Treatment Outcome
9.Influence and mechanism of He-Ne laser on scar formation of filtration canal after trabeculectomy in rabbit
Rui-Lin, WANG ; Gui-Gang, LI ; Zhi-Qi, CHEN ; Yan, XIANG ; Hong, ZHANG
International Eye Science 2010;10(9):1650-1653
AIM: To investigate the influence of He-Ne laser on connective tissue growth factor (CTGF) expression and collagen formation of fibroblast in filtration site after trabeculectomy in rabbit, and to discuss the mechanism for preventing scar formation with He-Ne laserin vivo.METHODS: The upper nasal limbus area next to the upper rectus muscle in right eyes Received 10 minutes He-Ne laser irradiation (200mW/cm2) every day for three days, the left eyes served as control. Twenty-four hours after the last irradiation, both eyes of the rabbits were took trabeculectomy surgery. The expressions of CTGF in the filtration area were tested on the 7th, 14th and 28th day after surgery and collagen density was tested on the 14th and 28th day after surgery. Each of the time point had 7 rabbits. RESULTS: The expression of CTGF was lower than that of the control group's on the 7th and 14th day after trabeculectomy surgery (P=0.01, P=0.005). When examined on the 14th and 28th day, the collagen density of irradiation group were significantly lower than that of the control group's (P=0.013, P=0.01).CONCLUSION: Pretreating the filtration area with 200mW/cm2 He-Ne laser may be helpful in preventing scar formation after trabeculectomy in rabbit, possibly due to downregulation of the expression of CTGF and collagen synthesis in fibroblasts. He-Ne laser may be developed into a new scar preventing method in filtration surgery.
10.Results of preservative free sodium hyaluronate and basic fibroblast growth factor eye drops in the treatment of dry eyes after ophthalmic surgery
Meng, WANG ; Gui-Qiu, ZHAO ; Shao-Xin, PAN ; Li-Li, ZHANG ; Kui-Xiang, LIU
International Eye Science 2008;8(10):1972-1974
AIM: To evaluate the results of preservative free sodium hyaluronate and basic fibroblast growth factor(bFGF) eye drops in the treatment of dry eyes after eye operation.·METHODS: One hundred and thirty two patients of dryeyes underwent 149 different eye surgeries. The Schirmer test Ⅰ and tear break up time (BUT) were performed prior to surgery and post operatively after 2 weeks and after 4 weeks when the patients were using Oflaxacine, Tobramycin-dexamethasone eye drops. In addition, preservative free sodium hyaluronate and bFGF eye drops were given post operatively. The antibiotic-cortisone eye drops were stopped after the requirted period depending on the type of eye surgery done, and preservative free sodium hyaluronate and bFGF eye drops were stopped after 4 weeks.Then, after 5 weeks Schirmer test Ⅰ and BUT were done.·RESULTS: The Schirmer test Ⅰ and BUT results prior to surgery and after eye surgery showed a statistically significant improvement of the dry eye condition.·CONCLUSION: Usage of preservative free sodium hysluronate and bFGF eye drops help definitely in the management of dry eye after ophthalmic surgery.