2.Prokaryotic expression and purification of human Smith D1 antigen
Wen-Bing WU ; Xiao-Peng LAN ; Xiang-Yue YANG ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.
3.Eukaryotic Expression and Primarily Application of Human Smith D1 Antigen in Methylotrophic Yeast Pichia pastoris
Xiang-Yue YANG ; Wen-Bing WU ; Xiao-Peng LAN ;
Microbiology 1992;0(01):-
To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.
4.Research advances of photosensitizer in photodynamic therapy of glioblastoma
Hong-cheng ZHAO ; Yue-qing WANG ; Qing-yun LI ; Hao DENG ; Xiao TAN ; Xiao-wen LIU
Acta Pharmaceutica Sinica 2022;57(6):1750-1757
Glioblastoma is a malignant tumor in central nervous system, which has strong invasion, poor prognosis and short survival time. At present, the main treatment strategy of glioblastoma is surgical excision, supplemented by radiotherapy and chemotherapy. However, due to incomplete resection and high recurrence rate, it is urgent to find novel therapeutic method for glioblastoma. Photodynamic therapy, as a promising non-surgical treatment, provides a new strategy for postoperative adjuvant therapy of glioblastoma. This review summarizes the mechanism and clinical application of photodynamic therapy mediated by various photosensitizers in glioblastoma, in order to provide help for the treatment of glioblastoma.
5.A randomized,double-blind,multicenter study of adefovir dipivoxii in treating patients with HBeAg-positive chronic hepatitis B
Hong ZHAO ; Chong-Wen SI ; Yue-Xin ZHANG ; Xin-Yue CHEN ; Lei WANG ; Xiao-Ping TANG ; Chunliang LEI ;
Chinese Journal of Infectious Diseases 2000;0(02):-
Objective To evaluate the efficacy and safety of adefovir dipivoxil(ADV)in treating patients with hepatitis B e antigen(HBeAg)positive chronic hepatitis B.Methods In this randomized,double blind,placebo-controlled,multicenter trial,210 eligible patients with HBeAg positive chronic hepatitis B were recruited and randomized(randomization ratio was 2:1)receiving ADV 10 mg/d for 48 weeks(ADV+ADV group,n=142)or placebo for 24 weeks followed by ADV 10 mg/d for 24 weeks(PLB+ADV group,n=68).The primary endpoint was virological response. The secondary endpoint was serologic response(HBeAg loss rate and HBeAg seroconversion rate) and alanine aminotransferase normalization rate.Results After 24 weeks therapy,mean reduction of hepatitis B virus(HBV)DNA level comparing with that of baseline was 3.12 log_(10)copy/mL in ADV +ADV group while it was 0.95 log_(10)copy/mL in PLB+ADV group.The percentages of patients with HBV DNA clearance(HBV DNA level
6.Efficacy of iris location to femtosecond-combined wavefront guided LASlK for myopia and astigmatism
Ke-Jie, LIN ; Jun, CHEN ; Wen, LIN ; Xiao-Dong, LIN ; Yue-Ming, ZHOU ; Liang-Ding, ZHENG
International Eye Science 2016;16(6):1056-1059
AIM: To observe effect of the iris location to femtosecond - combined wavefront guided LASIK for myopia and astigmatism.?METHODS:The patients with astigmatism >1. 0D during the same time and followed up for 1a were selected. A total of 129 eyes in 67 patients were treated under iris location with femtosecond-combined wavefront guided LASIK ( experimental group) and 161 eyes in 82 cases with femtosecond-combined wavefront guided LASIK ( control group) . Laser cutting went with the same laser machine. The uncorrected visual acuity ( UCVA ) , best corrected visual acuity ( BCVA) , and wavefront aberration between the two groups were compared at 1, 3, 6mo and 1a after surgery.?RESULTS:At 1 and 3mo after surgery, the number of patients with better postoperative UCVA than preoperative BCVA between the two group showed a statistically significant difference (χ2=6. 423, P=0. 011,χ2=14. 431, P=0. 01 ); at 1d and 1mo after surgery, the residual astigmatism showed a statistically significant difference between two groups (t=1. 98, P<0. 05; t=2. 23, P<0. 05). At 3, 6mo and 1a after surgery, the differences on the change of residual astigmatism between the two groups weren’t significant ( P > 0. 05 ). At 6mo and 1a after surgery, the differences on UCVA between the two groups weren’t significant ( P > 0. 05 ). Until 1a after surgery, the root mean square ( RMS ) of high order wavefront aberration of the two groups, spherical aberration and coma aberration ( COMA ) were all enhanced compared to before surgery(P<0. 05). At 1, 3mo after surgery, the RMS showed a statistically significant difference between two groups (P<0. 05). At 1, 3, 6mo, 1a after surgery, the increase of COMA in experimental group was significantly lower than that in control group (P<0. 05).? CONCLUSION: Iris location technology applied in femtosecond - combined wavefront guided LASIK for myopia and astigmatism, can make the vision recovery faster, the RMS of high order and COMA increase less, the residual astigmatism less, show better and more stable treatment effect.
7.Analysis on postoperative variation of corneal posterior surface heights after femtosecond LASIK and small incision lenticule extraction of high myopia
Yue-Ming, ZHOU ; Jun, CHEN ; Wen, LIN ; Xiao-Dong, LIN ; Ke-Jie, LIN ; Liang-Ding, ZHENG
International Eye Science 2016;16(10):1978-1980
AIM:To contrast analysis of postoperative variation of corneal posterior surface heights after Femtosecond LASlK ( FS - LASlK ) and small incision lenticule extraction ( SMlLE) for high myopia.
●METHODS: Sixty-seven cases of high myopic patients (132 eyes) operated with laser corneal refractive in our hospital from May to Dec. in 2015 (-6. 00D≤spherical equivalent degree≤-10. 00D) were selected and divided into FS-LASlK group and SMlLE group. The thickness of corneal flaps at FS-LASlK and the thickness of map at SMlLE were designed to be 110μm. Corneal posterior surface heights were examined by Pentacam at preoperation, postoperative 3 and 6mo after FS-LASlK and SMlLE operation. Surface height changes after preoperative, postoperative 3 and 6mo were compared by measuring Pentacam corneal analysis system.
●RESULTS: Six months after operation, the FS-LASlK posterior corneal surface height was 6. 47 ± 1. 65mm, significantly higher than 5. 20 ± 1. 32mm before operation. SMlLE posterior corneal surface height was 6. 40 ± 1. 33mm, significantly higher than 5. 18 ± 1. 25mm before operation, the differences were statistically significant( P<0. 05). Six month after surgery, two methods of corneal surface height variation obtained was 1. 29 ± 1. 28mm and 1. 22 ± 0. 89mm, there was no significant difference ( P>0. 05).
●CONCLUSION:After FS-LASlK and SMlLE, the corneal posterior surface is protrusive. FS - LASlK is slightly obvious than SMlLE in early period. The stability of the posterior surface is better after SMlLE.
8.Cyclic stress promotes the proliferation of rabbit condyle chondrocytes
Wen LIU ; Yan WANG ; Ling SONG ; Chunyan ZHANG ; Yue ZHANG ; Xiao YUAN
Chinese Journal of Tissue Engineering Research 2014;(20):3144-3148
BACKGROUND:The condyle is one of important areas of lower mandible growth, and has growth remodeling capacity lifetime. The in vivo functional research fails to obtain satisfactory outcome due to the complexity of physiological environment, nondirectiveness of stimulating factor transduction, and difficulty to control experimental conditions. Further studies wil explore the effect of stress on condyle chondrocytes in vitro.
OBJECTIVE:To observe the effect of cyclic stress on the proliferation of rabbit condyle chondrocytes cultured in vitro.
METHODS:Rabbit condyle chondrocytes were separated and cultured in vitro. Passage 3 chondrocytes were subject to cyclic tensile stress (10%surface elongation, 6 cycles/min) for 1, 6, 12, 24 hours. While those cells without stress served as the control group. The cellcycle changes were detected with flow cytometry analysis, and the cellproliferation were analyzed by MTT assay.
RESULTS AND CONCLUSION:The flow cytometry analysis showed that S-phase promoting factor exhibited significant differences at 6 and 12 hours compared with control group, and got the maximum value at 24 hours (P<0.01). MTT assay results showed that the cells proliferation at 6 and 12 hours exhibited significant differences compared with the control group, and got the maximum value at 24 hours (P<0.01). The cyclic stress can obviously promote the proliferation of condyle chondrocytes, and the stimulative effect can be sustained 24 hours.
9.Monitoring results of human brucellosis epidemic from 2008 to 2012 in Harbin
Jian-bin, CAO ; Xiao-li, GAO ; Bo, JIANG ; Yue, ZHAO ; Wen-cui, ZHANG
Chinese Journal of Endemiology 2013;32(5):512-515
Objective As brucellosis epidemic is constantly rising in Harbin in recent years,we analyze the epidemic and surveillance results at municipal surveillance spots of brucellosis from 2008 to 2012,to find out the epidemic characteristics of Harbin brucellosis,and to provide scientific evidences for development of countermeasures.Methods Brucellosis surveillance data which were collected from the National Diseases Surveillance Information Management System from 2008 to 2010,the epidemic characteristics of Harbin brucellosis (including regional,time and population distribution) and the municipal surveillance results in Harbin (including epidemiological investigation and serological surveillance) were analyzed.Results A total of 1348 cases of brucellosis were reported in Harbin from 2008 to 2012,and the incidence of brucellosis increased from 1.79 per 100 000 to 4.62 per 100 000; in addition to Pingfang District (2008-2011) and Fangzheng County(2009 and 2011),all eighteen districts (counties or cities) in Harbin had reported epidemic in the past five years,the highest incidence was in Yilan (382 cases),and the incidence was significantly increased in Shuangcheng(111 cases) and Binxian(73 cases) in 2012; the time of onset focused on March to July,which accounted for 62.54%(843/1348) of annual incidence; age distribution,mainly young adults aged 30 to 60(71.59%,965/1348); gender distribution predominantly male (75.45%,1017/1348) ; occupational distribution farmers(84.87%,1144/1348),and the number of infected students and house workers were also high in the past five years(4.23%,57/1348 and 2.52%,34/1348).In the municipal surveillance spots from 2008 to 2012,13 010 cases were surveyed,of which 2695 cases received serological test,the positive rate was 3.93% (106/2695),and the positive rate was increasing year by year.Conclusions The incidence of Harbin brucellosis is increasing year by year,onset season mainly in the spring and summer,young men with a high incidence,and farmers are main professional people.Brucellosis has also occurred among students,domestic workers,which should be vigilant.Therefore,we should further strengthen surveillance,timely analyze and report the epidemic,and provide a scientific basis to ensure each measure of eliminating the sources of infection put in place,vigorously develop health education,strengthen inter-departments cooperation,and control the spread of epidemic.
10.Modification by wheat germ agglutinin delays the ocular elimination of liposome.
Wen-Jian ZHANG ; Dong-Xiao YANG ; Ling-Lin FENG ; Fei WANG ; Gang WEI ; Wei-Yue LU
Acta Pharmaceutica Sinica 2014;49(4):543-549
The purpose of this study is to explore the feasibility of wheat germ agglutinin (WGA) modified liposome as a vehicle for ophthalmic administration. Liposome loaded with 5-carboxyfluorescein (FAM) was prepared by lipid film hydration method. WGA was thiolated and then conjugated to the surface of the liposome via polyethylene glycol linker to constitute the WGA-modified and FAM-loaded liposome (WGA-LS/FAM). The amount of thiol groups on each WGA molecule was determined, and the bioactivity of WGA was estimated after it was modified to the surface of liposome. The physical and chemical features of the WGA-modified liposome were characterized and the ocular bioadhesive performance was evaluated in rats. The result showed that each thiolated WGA molecule was conjugated with 1.32 thiol groups. WGA-LS/FAM had a mean size of (97.40 +/- 1.39) nm, with a polydispersity index of 0.23 +/- 0.01. The entrapment efficacy of FAM was about (2.95 +/- 0.21)%, and only 4% of FAM leaked out of the liposome in 24 h. Erythrocyte agglutination test indicated that after modification WGA preserved the binding activity to glycoprotein. The in vivo ocular elimination of WGA-LS/FAM fitted first-order kinetics, and the elimination rate was significantly slower than that of the unmodified liposome, demonstrating WGA-modified liposome is bioadhesive and suitable for ophthalmic administration.
Absorption, Physicochemical
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Adhesiveness
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Administration, Ophthalmic
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Animals
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Drug Carriers
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Eye
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metabolism
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Fluoresceins
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chemistry
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Liposomes
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administration & dosage
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chemistry
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pharmacokinetics
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Male
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Particle Size
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Polyethylene Glycols
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chemistry
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Rats
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Rats, Sprague-Dawley
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Wheat Germ Agglutinins
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administration & dosage
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chemistry
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pharmacokinetics