1.Cloning and expression of human glucagon-like peptide-1 cDNA
Zhi-Zhen ZHANG ; Ji-Fang MAO ; Hong DOU ; Sheng-Sheng YANG
Academic Journal of Second Military Medical University 2001;22(4):316-318
Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion: Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.
2.Cloning and expression of human glucagon-like peptide-1 cDNA
Zhi-Zhen ZHANG ; Ji-Fang MAO ; Hong DOU ; Sheng-Sheng YANG
Academic Journal of Second Military Medical University 2001;22(4):316-318
Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion: Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.
3.Relationship between the Ratio of Plasma Adrenomedullin/Endothelin-1 and Neuron-Specific Enolase in Full-Term Neonates with Hypoxic-Ischemic Encephalopathy
shi-fa, ZHANG ; ming-xiong, ZHOU ; shuang-gen, MAO ; chang-sheng, DOU ; guo-cheng, XU
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To explore the relationship of the ratio of plasma adrenomedullin(AM)and endothelin-1(ET-1)with serum concentration of neuron-specific enolase(NSE)in full-term neonates with hypoxic-ischemic encephalopathy(HIE).Methods Plasma concentrations of AM,ET-1 and serum NSE from 32 full-term neonates with HIE were detected by radioimmunoassay(RIA)on the 1,3 and 7 d after parturition,30 neonates in the corresponding periods in our hospital were employed as controls.The infants with HIE were divided into mild,moderate or severe group in terms of diagnostic standard of HIE.Results 1.Plasma concentrations of AM and ET-1 in newborns with mild,moderate or severe HIE were significantly higher than that of control group at 1 d after life with a decline from 3-7 d(Pa
4.Determination of Aliphatic Amines in Infant Food Packaging Materials by High Performance Liquid Chromatography
Cuncun SHENG ; Shijuan ZHANG ; Fangfang WANG ; Zhongyin JI ; Kun DOU ; Jinmao YOU
Chinese Journal of Analytical Chemistry 2016;44(9):1365-1371
Aliphatic amines in infant food packaging materials were extracted and concentrated by 0 . 5 mL of acidified methanol using gas purge microsyringe extraction ( GP-MSE ) . Pre-column fluorescence labeling of amines was achieved in mild conditions with 10-ethyl-acridine-2-sulfonyl chloride ( EASC ) as labeling reagent. The derivatization was carried out at 60℃ and pH 10. The derivatives were successfully separated on a Hypersil GOLD column with excitation and emission wavelengths of 262 and 430 nm, respectively. The detection limits were in the range of 0. 4-0. 6 μg/kg, and the quantitation limits were in the range of 1. 2-2. 1 μg/kg. All analytes were in good linearity in the concentration range of 2. 0-2000 μg/L with correlation coefficients of higher than 0. 998. The developed method was characterized by celerity, accuracy and high sensitivity. It was successfully applied to the determination of aliphatic amines in infant food packaging materials.
5.The expression and significance of decorin in chronic rejection of liver transplantion
Jianying SHANGGUAN ; Zhen-Yu TI ; Xiao LI ; Xiao-Jun HU ; Fu-Qin ZHANG ; Zhao-Sheng YONG ; Ke-Feng DOU ; Kai-Shan TAO ;
International Journal of Surgery 2010;37(11):739-742,封3
Objective To investigate expression and significance of decorin(DCN)in liver tissue and serum of liver transplant patients with chronic rejection(CR).Methods Immunohistochemistry(SP method)was used to detect expression of DCN in liver tissue of 16 normal controls, 20 patients with cirrhosis, 46 liver translantion patients without CR and 8 patients with CR.Enzyme-linked immunosorbent assay method(ELISA method)was used to determined the content of DCN in serum of all research subjects.Results The expression of DCN was negative in normal hepatic tissues and with/without CR, cirrhosis tissues showed strong expression of DCN.The positive expression rate and the average optical density value of DCN in liver transplant tissues with CR had significant difference comparing with Cirrhosis tissues(25% vs 55%, 0.1249 ±0.0039 vs 0.2357 ±0.0396, P <0.01,while no statistic siqnificance compared to normal liver tissues and those without CR.The level of DCN in serum was significantly higher in liver transplant patients with CR, with significant difference comparing with normal people, liver cirrhosis and transplant liver patients without CR(54.0833 ± 6.0325)μg/L vs(1.0232 ± 0.9105)μg/L,(12.6202 ± 1.5370)μg/L,(17.7102 ± 2.3562)μg/L, P < 0.01).The concentration of DCN in serum showed a positive correlation with the degree of CR.Conclusions DCN showed negative expression in liver tissue and increased significantly in serum of liver transplantation patients with CR.This suggests that DCN may be involved in occurrence and development of CR.At the same time the determination of DCN in serum maybe become an important indicator of the early diagnosis, development and prognosis of CR for liver transplant patients.
6.Values of high-frequency ultrasound, elastography and ultrasound contrast in diagnosis of thyroid solid nodules
Ning SHANG ; Hui DU ; sheng Li GUO ; Jun LI ; Na SHI ; yan Su ZHANG ; gui Dou ZHAO
Chinese Medical Equipment Journal 2017;38(9):67-70,77
Objective To investigate the diagnostic criteria of thyroid solid nodules by high-frequency ultrasound,elastography and ultrasound contrast.Methods Totally 87 patients with thyroid solid nodules underwent high-frequency ultrasound,elastography and ultrasound contrast before surgery,distinguishing benign from malignant lesions was executed based on pathological results,and according to the pathological character the image features by the three measures were compared.There were 40 patients divided into a control group going through the examination by high-frequency ultrasound,and 47 ones into an observation group with the examinations by the three measures.Pathological results were used as gold standard,and the two groups were compared on diagnostic sensitivity.Results High-frequency ultrasound showed significant differences of benign and malignant nodules in equal echo,very low echo,irregular peripheral halo,rates of clear nodule boundary and aspect ratio higher than as well as microcalcification (x2--4.603,17.394,5.833,14.832,10.107 and 27.392,and all Pvalues were less than 0.05).There were 85% benign nodules gained grade Ⅰ and Ⅱ elastography images,81.82%malignant nodules had grade Ⅲ and ⅣV elastography images,and there were significant differences between benign and malignant nodules (x2=53.42,P=0.00).Ultrasound contrast found annular enhancement in most of benign nodules (x2=39.16,P<0.01) while uneven enhancement in most of malignant nodules (x2=40.28,P<0.01),and the nodule ranges increasedsignificantly after contrast (t=2.19,P<0.05).The observation group had the sensitivity being 82.86%,which was statisticallyhigher than 59.38% in the control group (x2=4.537,P--0.033).Conclusion High-frequency ultrasound,elastography and ultrasound contrast all show the manifestations of thyroid solid nodules,and the comprehensive application of the above three measures contributes to determining the nature of the nodules.
7.Percutaneous radiofrequency ablation for liver cancer located under the diaphragm
Yu-Xuan WU ; Yong-Chong DOU ; Yan-Fang ZHANG ; Xin-Yin SHENG ; Xue-Feng ZHENG ; Guo-Ping SUN ; Jian-Ming XU ;
Journal of Interventional Radiology 2006;0(12):-
Objective To evaluate the feasibility and safety in the treatment of liver cancer located under the diaphragm with cool-tip radiofrequency ablation(RFA)percutaneously under CT guidance.Methods 20 patients with total 25 lesions were treated by CT-guided RFA with cool-tip electrode involving the induced necroses.The postoperative efficacy was evaluated by enhanced CT or MRI.Results 72% lesions were completely necrotized(18/25),28% lesions were majorly necrotized(7/25).No severe complications occurred. Conclusion CT-RFA with cool-tip electrode is effective and safe in treating liver cancer located under the diaphragm.
8.Correlation analysis of chronic hepatitis C with nodular goiter.
Dongxu JIANG ; Chong ZHANG ; Yang DING ; Qiuju SHENG ; Jingyan WANG ; Xiaoguang DOU
Chinese Journal of Hepatology 2015;23(3):175-179
OBJECTIVETo assess the relationship between nodular goiter and hepatitis C virus infection.
METHODSNinety-seven cases of early treatment in patients with chronic hepatitis C were collected for analysis.Data on patient age,sex,hepatitis duration and other general information were collected.In addition, data on clinical measures of thyroid function (including T3, t4, tSH) and thyroid autoantibodies (thyroid peroxidase antibody TPO-Ab, thyroglobulin antibody Tg-Ab), as well as findings from thyroid dimensional ultrasonography were collected. One hundred and eleven cases of early treatment in patients with chronic hepatitis B and 106 eases of females 40 years old or older with high risk of nodular goiter were collected for use as controls.The relationship between nodular goiter with thyroid function, thyroid autoantibodies levels,sex,age,and hepatitis C virus infection were statistically analyzed.
RESULTSThe prevalence rates of nodular goiter in the chronic hepatitis C group, the chronic hepatitis B group and the more than or equal to 40 year-old women with high risk of nodular goiter were 53.6%,36.9% and 59.4% respectively.The prevalence rates of nodular goiter in the chronic hepatitis C group and the more than or equal to 40 year-old women with high risk of nodular goiter were significantly higber than that in the chronic hepatitis B group (x² values: 5.820 and 10.996, P < 0.05). The average age of patients with chronic hepatitis C combined with nodular goiter was significantly higher than their counterparts without goiter (F=6.408, P < 0.05),and the prevalence rate in the more than or equal to 40 year-old women with high risk of nodular goiter was significantly higher than that of their counterparts who were less than 40 years-old (60.0% vs. 23.5%; x² =7.499, P less than 0.05). The prevalence of nodular goiter in patients with chronic hepatitis C was significantly greater for females than for males (62.1% vs. 41.0%; x 2 =4.152, P < 0.05).The prevalence of nodular goiter in patients with chronic hepatitis C was also significantly higher for females more than or equal to 40 years old than for males (70.2%, 33/47 vs. 45.5%,15/33; x² = 4.952, P < 0.05).The duration of hepatitis, thyroid function and thyroid autoantibodies were similar between the patients in the chronic hepatitis C group with or without nodular goiter.
CONCLUSIONSThe patients with chronic hepatitis C had a higher prevalence of nodular goiter,with an average of up to 53.6%, than the patients with chronic hepatitis B,and the women the more than or equal to 40 years old had even higher prevalence, at 70.2%, suggesting that patients with chronic hepatitis C should be routinely examined by thyroid ultrasound. Thyroid function and thyroid autoantibodies were not correlated with prevalence of goiter among the chronic hepatitis C patients.
Autoantibodies ; Female ; Goiter, Nodular ; Hepatitis B, Chronic ; Hepatitis C, Chronic ; Humans ; Male ; Prevalence
9.Identification of impurity peaks in the HPLC chromatogram by LC-MS and two-dimensional chromatographic correlation spectroscopy.
Zhen-Zhen CHEN ; Dou-Sheng ZHANG ; Nan WANG ; Fang FENG ; Chang-Qin HU
Acta Pharmaceutica Sinica 2012;47(4):492-497
A novel qualitative analytical method by using two-dimensional chromatographic correlation spectroscopy techniques for recognizing impurity peaks of HPLC methods of quality control and LC-MS chromatographic system was established. The structures of major degradation products of ceftizoxime and cefdinir were identified by LC-MS and MassWorks application; the standard chromatographic and spectral data of the degradation impurities were obtained by high-performance liquid chromatography with diode array detection. The impurity peaks of two-dimensional chromatography were matched by comparison of spectra and calculating correlation coefficients. Peaks in chromatography can be identified accurately and rapidly in different chromatographic systems such as column and mobile phase changed. The method provides a new way and thought to identify the peaks in quality control of impurities without reference impurity substances.
Ceftizoxime
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analysis
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chemistry
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Cephalosporins
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analysis
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chemistry
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Chromatography, High Pressure Liquid
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methods
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Chromatography, Liquid
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methods
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Drug Contamination
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Mass Spectrometry
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methods
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Quality Control
10.Closed reduction and percutaneous Kirschner-wire fixation for the treatment of supracondylar fractures in children: a report of 37 cases.
Nai-qiang WEI ; Fu-dou YIN ; Lin-lin ZHANG ; Jing-hui GENG ; Wei-chao SHENG
China Journal of Orthopaedics and Traumatology 2008;21(4):309-310
Bone Wires
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Child
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Child, Preschool
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External Fixators
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Female
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Fracture Fixation
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methods
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Humans
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Humeral Fractures
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surgery
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Male