1.Separation of SO-Rb_ (50)cells antigen corresponding to the monoclonal antibody of anti-retinoblastoma
Fangwei YING ; Yongping LI ; Song TANG
Chinese Journal of Ocular Fundus Diseases 2001;0(03):-
Objective To separate the SO-Rb 50cells antigen corresponding to the monoclonal antibody of anti-retinoblastoma. Methods The antigen corresponding to the monoclonal antibody of anti-retinoblastoma was separated elementarily by ion-exchange chromatography, and was identified by dot-blotting using the monoclonal antibody of anti-retinoblastoma. The target protein band of the antigen was separated in light of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results A special unmixed band of SO-Rb 50cells antigen was separated with the relative molecular weight of 83?103. Conclusion The antigen corresponding to the monoclonal antibody of anti-retinoblastoma could be separated from SO-Rb 50cells.
2.Establishment of Experimental Diabetic Nephropathy Rats Model: Optimum Selection Ⅱ
Ying XU ; Shiwen ZHOU ; Jianlin TANG ; Yongping HUANG ; Sha CHEN
China Pharmacy 2005;0(19):-
OBJECTIVE: To compare the effects of different factor combinations on diabetic nephropathy(DN)rats model and to determine the preferred plan for the establishment of DN rats model.METHODS:A total of 30 SD rats were randomly divided into 5 group(6 rats in each group):group A was assigned to receive placebo(normal control),group B to receive high-lipids and high-sucrose feeding,group C to receive high-lipids and high-sucrose feeding plus streptozotocin(STZ),group D to receive high-lipids and high-sucrose feeding plus doxorubicin hydrochloride and STZ,group E to receive high-lipids and high-sucrose feeding plus mononephroctomy and STZ 12 weeks later the model rats were put to death for the determination of serum parameters including blood lipid,glycosylated hemoglobin,renal function,aldose reductose(AR),glomerular filtration rate(GFR),SOD and MDA .RESULTS:The rats in group E showed significant increase in blood lipid,in particular the increase of AR,and which also showed significant differences in GFR and oxygen free radical indices as compared with normal group and the other model groups.CONCLUSIONS:The rats model that fed with high-lipids and high-sucrose feeding for 4wk prior to mononephroctomy followed by the postoperative intraperitoneal injection of low dose of STZ 2 weeks later proves to be the optimal one.
3.Study for gene mutation detection of circulating DNA with ligase-ELISA reaction
Haizhong CUI ; Na XIAO ; Yongping ZHANG ; Dagui CHEN ; Yitong TANG
Tianjin Medical Journal 2015;43(5):533-536
Objective To establish a single nucleotide polymorphisms genotyping (SNP) method for a convenient, accurate, and routine analysis of clinical samples. Methods Based on the design of oligonucleotide probe, the assay was performed through three steps:the conjunction of the detection probe, universal amplification, labeling and ELISA reaction. The genotype of each SNP was revealed by reading signals of each set of reaction tubes. This assay was applied to detect sixty-two plasma samples of lung cancer for circulating DNA for three SNPs of EGFR, c.2573T>G(L858R), EGFR, c.2582T>G>T(G719C). Results were compared with those obtained by direct sequencing. Results The heterozygote mutation was identified for L858R by both methods, although no mutation was detected for L861Q and G719C. Six samples were identified as heterozygotes with the new method, and only two samples were unambiguously identified as heterozygotes by the direct sequencing. Two additional samples could not be identified as heterozygotes because the peak of mutant allele was very low compared with that of wild allele. Conclusion The developed method enabled accurate identification of SNP in a convenient manner, and which is adapted to routine analysis from heterogeneous samples unambiguously.
4.Clinical Application of Novel Colloidal Gold Immunochromatographic Test Strip for Group B Streptococci
Yuxia WU ; Bin WU ; Xiuhua PAN ; Yongping TANG ; Banglao XU
Journal of Modern Laboratory Medicine 2015;(4):53-55
Objective To explore the effect of a novel colloidal gold immunochromatographic test strip for detection of group B streptococci (GBS).Methods A total of 202 cases of swab of vagina or neck of uterus were collected,and they were detec-ted by novel strip and control strip to evaluate their clinical applications.Results Sensitivity of novel strip was about 105 CFU/ml and the detection time was about 5 to 8 minutes,and it showed better sensitivity and shorter detection time com-pared with control strip.In the 202 cases of clinical samples,the detection results of 197 cases were in consistent with the control strip,however,the detection results of 5 cases were not in consistent.The positive coincidence rate and negative coin-cidence rate were 97.5% and 97.54% respectively,and the total coincidence rate and Kappa value were 97.52% and 0.948 respectively.The consistency test showed no significant difference between this strip and control strip.Conclusion This method was a effective technology for diagnosing of infection caused by GBS,and had high value in clinical application.
5.The value of biliary tumor markers for differentiatial diagnosis of benign and malignant biliary diseases
Lixin TANG ; Xu REN ; Lingling ZHANG ; Xiufen TANG ; Chunlan ZHU ; Yongping QU
Chinese Journal of Digestive Endoscopy 2014;31(1):22-25
Objective To investigate the value of biliary tumor markers for differential diagnosis of the benign and malignant biliary tract diseases.Methods Tumor markers (CA19-9,CEA and CA242) examination and bacterial culture were performed in a total of 160 patients,who underwent therapeutic endoscopic retrograde cholangiopancreatography (ERCP) for biliary diseases.Resuts There were significant differences between malignant group and benign group in bile and serum in the level of CA19-9,CEA and CA242 (P <0.05) ; Cut-off value,according to ROC curve,was 239 ku/l in CA19-9,40 ng/ml in CEA and 60 ku/ml in CA242,respectively.There were significant differences between the bile marker and the serum marker in sensitivity,accuracy,negative predicative value of CEA (P < 0.05).No significant differences was found in specificity between the serum group and the bile group.There were significant differences in bile CA19-9 level between cholangiocarcinoma,pancreatic cancer,duodenal papilla carcinoma with carcinoma metastasizing to bile duct,and hepatocellular carcinoma (P < 0.05).Both in benign group and malignant group,there were significant differences in CA19-9 level between infectious bile and noninfectious bile (P < 0.05).Conclusion The level of CA19-9,CEA and CA242 in bile can be applied to differentiate benign and malignant biliary diseases.The bile tumor markers do not have advantage over serum tumor markers in specificity for diagnosis.Bile bacterial infection can result in the elevation of bile CA19-9 while it does not have impact on differential diagnosis.
6.Establishment of the technique for the real-time fluorescence quantitative reverse transcription polymerase chain reaction by DNA melting curve analysis for detecting the CDR3 skewing of TCR alpha gene repertoire in the human peripheral blood
Xianying TANG ; Yongping SUN ; Rui MA ; Hongqian ZHU ; Zuguo TIAN ; Wanbang SUN ; Xinsheng YAO
Chinese Journal of Immunology 2000;0(08):-
Objective:To establish the technique for real-time fluorescence quantitative reverse transcription polymerase chain reaction(FQ-PCR)by DNA melting curve analysis for detecting the CDR3 shewing of TCR alpha gene repertoire in human peripheral blood.Methods:Total RNA of peripheral blood mononuclear cell(PBMC)from 4 healthy donors and 2 patients with lymphomatous leukemia were transcripted reversely into cDNA.The cDNA of 32 TRAV gene family CDR3 was amplified by FQ-PCR.Analysis of the monoclonal/oligoclonal/polyclonal CDR3 spectratyping with DNA melting curve.Results:The FQ-PCR products of 32 TRAV family CDR3 were showedas a blur land at the predicted of products size in healthy donors and parts of TRAV family CDR3 products disappeared in patients on 1.5% agarose gel by Gold-View staining.The 32 TRAV family CDR3 were showed with different frequencies by relative fluorescence quantitative in healthy donors and the patients.The CDR3 spetratyping for 32 TRAV families was showed as polyclonal peak(Gaussian distribution)in healthy donors but showed as different monoclonal/oligoclonal/polyclonal peak in the patients with lymphomatous leukemia with DNA melting curve analysis(we called "melting curve spectratyping of CDR3")Conclusion:The study suggests that the technique of "FQ-PCR with DNA melting curve analysis be convenience and celerity for detecting the CDR3 skewing of TCR alpha gene repertoire in human peripheral blood.
7.Effects of N-acetylcysteine on Alcoholism in Mice
Yongping HUANG ; Shiwen ZHOU ; Ying XU ; Jianlin TANG ; Lian WANG ; Jinyuan ZHANG ; Xue YANG
China Pharmacy 2001;0(07):-
OBJECTIVE:To investigate the effects of N-acetylcysteine(NAC)on the concentration of ethanol in serum and the activities of alcohol dehydrogenase in liver and stomach tissue in alcoholic mice.METHODS:All mice were given p.o white spirit30min after p.o saline or5%NAC solution,and the durations from the disappear of righting reflex to recovery in mice were recorded and the number of dead mice in24hours were countered;With the same procedure for six days,the mice were killed by withdrawaling blood from orbit,and taken out the liver and stomach immediately,the concentrations of ethanol in serum and the activities of alcohol dehydrogenase in liver and stomach were measured by chromatometry and colorimetric method respectively.RESULTS:In drunk test,5%NAC solution significantly postponed the time from p.o white spirit to the disappear of righting reflex(P
8.The effects of vitamin D deficiency on the cardiac oxidative stress in mice and the underlying mechanisms
Yi TANG ; Jing LIU ; Liang XIE ; Lijun WANG ; Qigao ZHANG ; Yongping PENG ; Jianbin GONG ; Shisen JIANG
Journal of Medical Postgraduates 2016;29(10):1050-1054
Objective Previous studies have shown that vitamin D deficiency is closely related to cardiac remodeling. How?ever, the underlying mechanisms have not been fully elucidated. Moreover, oxidative stress plays an important role on the pathologies of cardiac remodeling. The aim of this study was to explore the influence of VD deficiency on cardiac oxidative stress and the potential sig?nal pathway. Methods The male C57 mice ( 3 weeks old) were randomly divided into three groups: vitamin D deficiency ( VDD ) group ( vitamin D deficiency feed for 10 weeks) , vitamin D deficiency ( VDA) group ( vitamin D sufficiency feed for 10 weeks) and VDD+calcitriol ( CAL) group ( vitamin D deficiency feed for 10 weeks and then vitamin D sufficiency feed and calcitriol treatment for 10 weeks) . Results There were significant differences between the VDD group and the VDA group in the left ventricular end?diastolic diameter and left ventricular mass index (3.82±0.125 mm vs 3.748±0.092 mm, P<0.05) (119.30±8.54 vs 97.60±3.65, P<0.05). The number of myocardial cells stained with 8?OHDG was higher in the VDD group compared with the VDA group ( 65. 4 ± 2. 3 vs 21. 8 ± 1. 6, P<0.05) whereas was lower after calcitriol supplement. Furthermore, the expression of thioredoxin interacting protein (TXNIP) was sig?nificantly up?regulated and the ratio of p?ASK?1/ASK?1, cytochrome C release, and caspase3 activation were increased in the VDD group . Conclusion VDD can lead to cardiac oxidative injury and the up?regulation of TXNIP and the activation of ASK?1 related apoptotic signal cascade may be involved in this procedure.
9.Optimizing the Extraction of Alkaloids from Rhizoma Coptidis by Orthogonal Design
Yi YING ; Zhihong HE ; Shiwen ZHOU ; Jianlin TANG ; Yongping HUANG ; Xue YANG
China Pharmacy 2001;0(09):-
OBJECTIVE:To study the optimized process for extraction of the alkaloids from Rhizoma Coptidis.METHODS:Orthogonal design was used for studying the effect of alcohol concentration,volume,extraction time and times on the content of jatrorrhizine,epiberberine,coptisine,palmatine and berberine.RESULTS:The optimum extraction process is as follows:8 times of 50% alcohol,extracted for 4 times,each time for 2 hours.CONCLUSION:This extraction process is workable and reproducible,and can provide theoretic support for mass extraction of the alkaloids from Rhizoma Coptidis.
10.A study on the use of Peabody's fine motor development scale
Banghui LI ; Qi WANG ; Xiaoxi LUO ; Lijun ZHANG ; Qiuju TANG ; Yongping REN ; Nong XIAO ; Xueqin HOU
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(10):751-753
Objective To assess if evaluating with Peabody's fine motor development scale with 4 degree grading is more sensitive than with 3 degree grading, and whether or not it is feasible to evaluate by quantization with monthly averages. Methods A total of 864 normal children aged 1 month to 60 months were evaluated with the Peabody scale using 4 degree grading and 3 degree grading. The development results were averaged by month to express the development. Results Both ways, the monthly averages of children 4-9 months old were higher than the others. The values obtained with 4 degree grading were lower than those with 3 degree grading in each functional area, and the difference was more obvious with increasing age, but the differences were not statistically significant. With 3 degree grading the total score was equal to the actual score after the age of 9 months, but with 4 degree grading this was not true until at least 18 months. Conclusions Evaluating with Peabody's fine motor scale with 4 degree grading and quantization using monthly averages is reliable and more sensitive than 3 degree grading.