2.Integration and Expression of BGL1 Gene from Saccharomy-copsis fibuligera in Industrial Saccharomyces cerevisiae
Liang ZHANG ; Yan ZHOU ; Gui-Yang SHI ;
Microbiology 1992;0(03):-
The recombinant plasmid pPIC-gpd-bgl-hyg was constructed, which contained GPD2 promotor and terminator from industrial yeasts Saccharomyces cerevisiae, ?-glucosidase gene (BGL1) from Sac-charomycopsis fibuligera and hyg from hygromycin as the selected marker. With the yeast’s high efficiency of homologous integrated, the BGL1 gene was successfully integrated into industrial yeasts S. cerevisiae. The recombined yeast could grow on the cultures with the cellobiose as a sole carbon source, and the ?-glucosidase activity achieved 0.764 U/mL after 48 hours’ cultivation. In the experiments of VHG ethanol fermentation, the cellobiose concentration in broth of recombined yeast was 80% lower than that of indus-trial yeast.
3.Accuracy of intraocular lens power prediction by using six different formulas
Yan JIANG ; Yuying SHI ; Wenli YANG
Ophthalmology in China 1994;0(02):-
Objective To compare the accuracy of intraocular lens(IOL)power calculations by using 6 formulas(SRK-T,Haigis,Binkhorst-Ⅱ,Hoffer Q,Holladay-1,SRK-Ⅱ)in different axial lengths of the eye,in order to improve the accuracy of predicating IOL powers for patients after cataract surgery.Design Retrospective comparative clinical analysis.Participants 169 patients(169 eyes)with age-related cataract underwent regular phacoemulsification and posterior chamber IOL implantation.Methods 169 eyes with cataract were divided into five groups according to the axial lengths:AXL≤22mm,22mm28.4mm.The axial lengths and keratometry of the 169 eyes were measured with A-mode ultrasound before operation,and the actual postoperative refraction was measured with the methods of computer and retinoscopy optometry when patients could gain best corrected visual acuity one month after surgery or later.Inputting the axial lengths,keratometry and IOL powers into A-mode ultrasound,the predicted postoperative refraction was calculated with SRK-T,Haigis,Binkhorst-Ⅱ,Hoffer Q,Holladay-1,SRK-Ⅱ formulas,and the differences of predicted and actual postoperative refractions in different axial lengths were compared.Main Outcome Measures The absolute value with the differences of the predicted and actual postoperative refractions,i.e.absolute predicted errors.Results(1)Comparing the difference of predicted and actual postoperative refraction,there was no statistical significance between the five formulas in the groups of axial length(AXL)≤22mm,22mm28.4mm,but the average absolute predicted error of Haigis and SRK-T formulas is significantly smaller than SRK-Ⅱand Binkhorst-Ⅱ formulas.Conclusions In existing sample size,there was no obvious influence on postoperative refraction when using other five formulas except Binkhorst-Ⅱ.But in the extremely long AXL group,Haigis and SRK-T formulas have some ascendancy over the other formulas.
4.Determination of 17 types of phthalates in drinking water by solid-phase extraction coupled with multiple reaction monitoring of gas chromatography-mass spectrometry
SHI Mengxing ; YANG Yan ; DU Limin
Journal of Preventive Medicine 2025;37(1):102-108
Objective:
To determine 17 types of phthalic acid esters (PAEs) in drinking water using solid-phase extraction coupled with gas chromatography-mass spectrometry (GC-MS) in multiple reaction monitoring (MRM) mode.
Methods:
One litre of commercially available bottled water was purified using an HLB solid-phase extraction column, and was eluted with ethyl acetate, dichloromethane and average methanol. Seventeen types of PAEs were detected using a triple quadrupole gas chromatography-mass spectrometer in MRM mode. By optimizing the temperature programming and adjusting the mass spectrometry collision energy, collection efficiency was improved and matrix interference was reduced. The precision and accuracy of this method were assessed by determining the standard curves, detection limits, quantification limits, relative standard deviations (RSD) and average spiked recovery rates for the 17 types of PAEs.
Results:
The 17 types of PAEs showed good linear relationships between mass concentration and chromatographic peak areas in the range of 0.02 to 1.0 mg/L, with correlation coefficients all greater than 0.999 1. The detection limits ranged from 0.002 9 to 0.009 7 mg/kg, the quantification limits ranged from 0.008 7 to 0.029 1 mg/kg, the RSD ranged from 0.8% to 3.0%, and the average spiked recovery rates ranged from 88.8% to 111.8%.
Conclusion
Solid-phase extraction coupled with MRM of GC-MS can better determine low concentrations of PAEs in drinking water.
5.Study of specific immune response on mice induced by a DNA vaccine co-expressing both MAGE-1 and interleukin-18
Guangmin YANG ; Yang SHI ; Yan TAN ; Shouqing LI ; Yinfu MA
Chinese Journal of Immunology 2001;0(07):-
Objective:To observate immunologic response of immunized mice by the vaccine pcIL-18-MAGE.Methods:Abundant plasmids were extracted and immunized mice,two booster injection were carried out at ten days intervals.Seven days after the third immunization,serum antibody against MAGE-1 and T cell subgroup were detected by FCM,and CTL killing activity were assayed through MTT.Results:About serum antibody against MAGE-1 and T cell subgroup,recombinant plasmids groups were all higher than the vacant plasmid pcDNA3 group(P
6.Effect of EDFS40 on morphology and vasomotoricity of rabbit carotids in cryopreservation
Tao SHI ; Yi Lü ; Yang YAN ; Xigang GENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2009;30(6):665-668
Objective To assess the effect of EDFS40 on the structure and vasomotoricity of rabbit carotids in cryopreservation. Methods The fresh carotids, taken from rabbits under aseptic conditions, were preserved for 14 days in liquid nitrogen by vitrification or slow freezing method with both fresh and thawed carotids were observed by visual examination, optical and electron microscopes, respectively. Their endothelium-independent vasoconstriction and vasodilatation were detected in organ bath. Results The morphological changes of the EDFS40-vitrified arteries were similar to those of the fresh carotids under macrography, optical and electron microscopes. And there was no obvious difference in contraction to KCl between the cryopreservation group and the slow-freezing group (P>0.05). The contractile and relaxant responses of vitrified arteries to NE or SNP were significantly better than those of the frozen arteries (P<0.05); both, however, were still inferior to the vasomotoricity of the fresh arteries (P<0.05). Conclusion The rabbit carotids vitrified with EDFS40 can have their structure and vasomotor function maintained without obvious damage, which may indicate the usage of EDFS40 in vitrification of blood vessels.
7.MRI diagnosis of ejaculatory duct obstruction
Yan GUO ; Yaoping SHI ; Dong YANG ; Huanjun WANG ; Bitao PAN
Chinese Journal of Radiology 2012;46(7):633-635
Objective To explore the MRI features of ejaculatory duct obstruction.Methods During January 2003 to Dccember 2010,transrectal ultrasonography (TRUS) was performed for 106 patients and underwent surgical treatment for ejaculatory duct obstruction.Among them,16 patients underwent MRI examination.The MRI features of ejaculatory duct obstruction in these patients were summarized.Results Ejaculatory duct cysts,ranging in size from 4 mm ×4 mm ×7 mm to 4 mm ×4 mm ×9 mm and locating in the paramedian line,were detected in 5 of the 16 patients; ejaculatory duct dilation located in the paramedian line was detected in 7 patients,with the internal diameter of 5 to 30 mm. After contrast injection,significant enhancement of the wall of the ejaculatory duct was observed in 2 patients.Mullerian duct cysts complicated with dilated ejaculatory duct and seminal vesicles were detected in 4 patients,in whom the cysts were located in the median line,ranging in size from 4 mm × 5 mm × 6 mm to 34 mm×35 mm ×44 mm,with inverted teardrop shaped pointing toward the seminal colliculus.ConclusionThe most common MRI features of ejaculatory duct obstruction are ejaculatory duct dilation and ejaculatory duct cysts.
8.MR imaging features of adrenal rest tumor
Chen SU ; Yan GUO ; Wenjuan WANG ; Yaoping SHI ; Dong YANG
Chinese Journal of Radiology 2012;46(8):705-707
Objective To investigate the imaging features of adrenal rest tumor.Methods Twelve patients of adrenal rest tumor proved by surgery or clinical diagnosis were retrospectively analyzed.Among these 12 patients,12 were examined with ultrasound,11 with MR and 1 with CT. MR and CT were performed without and with intravenous injection of contrast material.The imaging features of adrenal rest tumor were retrospectively summarized and the relevant literatures reviewed. Results The adrenal rest tumors were found in testis in 10 of the 12 patients,and in ovaries and broad ligament in the remaining two.The imaging features of the testicular adrenal rest tumor were summarized as following:all patients had bilateral testicular masses without change of the testicular contour. On ultrasonography,the lesions were hypoechoic, with some hyperechoic areas and appeared highly vascularized on Colour Doppler ultrasonography.The masses showed iso-density on plain CT,and avid enhancement on post-contrast CT images.The masses ranging in size from0.7 cm×1.0 cm×2.2 cm to 2.3 cm ×2.7 cm ×2.9 cm with uniform signal intensity,lobulated margin on MRI.They exhibited iso- or slight hyperintensity on T1 WI and hypointensity on T2WI relative to normal testicular parenchyma.The tumors showed intense enhancement on post-contrast MR images. No abnormality was detected with Colour Doppler uhrasonography and MR in 2 patients of adrenal rest tumor in ovaries and broad ligament. Conclusion Combining imaging features with the typical clinical history,the diagnosis of adrenal rest tumor could be suggested pre-operatively.
9.Diagnostic value of endoscopic ultrasonography for ulcerative colitis
Ping YIN ; Zhenhua YANG ; Aoshuang HUANG ; Fei SHI ; Yan MAO
Chinese Journal of Digestive Endoscopy 2012;23(1):6-10
Objective To study the consistency and complementarity of endoscopic ultrasonography (EUS),white light endoscopy (WLE) and magnifying endoscopy (ME) in diagnosis of ulcerative colitis (UC).Methods We collected 125 cases of UC patients diagnosed by WLE and EUS (including 51 cases of WLE + ME + EUS).According to UC mucosal morphology under WLE and crypt openings under ME,we divided all the cases into several groups and analyzed intestinal wall thickness (TWT) under EUS in each group.Results According to the results of UC inflammation degree under WLE,all patients were divided into four groups: 16 severe cases,46 moderate cases,44 mild,and 19 remission stage.TWT results were (5.903 ± 1.551 ) mm,(4.673 ± 1.235 ) mm,(3.756 ± 1.322 )mm and ( 3.464 ± 0.970) mm,respectively.Differences were significant between any two groups ( P < 0.05 ),except for that between mild and remission groups.According to the results of UC inflammation degree under ME,all patients were divided into six groups: 9 cases of villous-like structure,9 cases of typical coral reef-like structure,8 severe coral reef-like structure,13 regular crypt opening,6 epithelial minimal defect and 6 small yellow spot (SYS).TWT results were (5.701 ±0.941 )mm,(5.518 ±0.581 )mm,(5.181 ±0.751 )mm,(3.763 ±0.659) mm,(3.587 ±0.461 )mm and (2.505 ± 0.330 )mm,respectively.Differences were significant between any two groups ( P < 0.05 ) except for those between epithelial minimal defect and regular crypt opening,typical coral reeflike structure,villous-like and severe coral reef-like structure.EUS results showed SYS (6/6) and regular crypt opening ( 10/13 ) were mostly located in mucosa,while lesions of severe coral reef-like structure (8/8) invaded the muscularis propria.Conclusion EUS shows high consistency with WLE and ME in diagnosis of UC inflammation degree and invasive depth.It could assist and even substitute ME for evaluation.
10.The study on levofloxacin resistance and gyrA gene mutation of Helicobacter pylori in Jinhua Zhejiang province
Li SHI ; Yan-ping CHENG ; Wei HUANG ; Ning-ming YANG ;
Chinese Journal of Digestion 2010;30(11):832-835
Objective To analyze levofloxacin resistance in Helicobacter pylori (Hp) and the sequence difference of gyrA gene in levofloxacin resistance and sensitive Hp strains. To explore the function of gyrA gene mutation in the development of levofloxacin resistance Hp strain.Methods From July 2007 to December 2008 in Department of Gastroenterology, Jinhua People hospital of Zhejiang Province, the gastric mucosa from gastroscopy biopsy of chronic gastritis and peptic ulcer patients were cultured in Hp selective medium under microaerobic condition at 37 degrees for three to five days. The Hp strains were isolated and identified by oxidase test, catalase test,urease test and UreA gene detecting. The levofloxacin susceptibility was determined by E-test and then resistance and sensitive strains were screened. The genomic DNA of Hp strains was isolated. The gyrA gene was amplified by PCR and the sequences were analyzed. Results 38 clinical isolated Hp strains were passed the levofloxacin susceptibility E-test, among those the minimum inhibitory concentration (MIC) of 12 strains was over 1.0 μg/ml, the percentage of resistant strain was 31.58%, while the sensitive stains was 68.42%. The gyrA gene sequence result indicated 10 resistant strain with 261, 271 and 272 10 site mutation, 2 strains with C261A mutation, one strain with C261G mutation, two strains with G271A mutation, 2 strains with A272G mutation, 2 strains with C261A,G271T andA272G mutation, 1 strain with C261G and A272G mutation. However, no mutation sites were found in 26 sensitive strains. Conclusion The rate of levofloxacir-resistance in isolated clinical Hp strain was high. The drug-resistance was associated with 261,271 and 272. site mutation of gyrA gene.