1.Development and applications of inexpensive genotyping methods
Journal of China Pharmaceutical University 2009;40(3):193-199
Single nucleotide polymorphism( SNP) is a major source of genetic differences, and the number of SNP is extremely large. Consequently, a high-throughput, rapid and cost-effective technique is essential for SNP typing. Recently, a set of solutions aimed directly at the bottlenecks in SNP detection were put forward, which including direct PCR from human whole blood, adapter-ligation mediated allele-specific amplification, gene polymorphism detection in one tube, microplate array parallel gel electrophoresis and microchip electrophoresis. These methods have been successfully applied in pharmacogenomics, disease-related research, herbal medicine identification and quality control. The results showed that the advantages of the genotyping platforms are highly specific, sensitive, easy to operate and inexpensive with a good prospect of application. The principles and applications of the genotyping platforms in detail were described in this paper.
2.Analysis of clinical effect for children with obstructive sleep apnea hypopnea syndrome
Chinese Journal of Primary Medicine and Pharmacy 2008;15(10):1623-1624
Objective To investigate the causes and therapeutic method for children with obstructive sleep apnea hypopnea syndrome. Methods 72 patients with obstructive sleep apnea hypopnea syndrome were treated by adenctonsillectomy, operation and precaution of postoperation complication. Results 62 patients (86.1% )were cure.8 patients(11.1% ) were better. 2 patients(2.7% ) were no change. Conclusion The children with obstructive sleep apnea hypopnea syndrome caused by glandular organ and hypertrophyoftonsil, can improve the surgical results and late results.
3.Genotyping three human SNPs by Cy5-ddNTP primer extension
Journal of China Pharmaceutical University 2006;(6):569-572
To establish a simple and rapid method to genotype the human single nucleotide polymorphisms(SNPs). Methods:In the presence of the appropriate Cy5-ddNTPs and dNTPs,the primer was extended by one or more bases depending on the sequence at the polymorphic site. The microchip electrophoresis was used to detect the product of extension.The switching characteristics of the primer extension reaction was increased by introducing an artificially mismatched base into the third position from the 3'-terminus of the primers. Results:Three human SNPs were detected and the alleles were distinguished on the basis of size of the extended product. All samples were accurately assayed within 100 s in microchip electrophoresis. Conclusion:This technique offers a high-speed,easy and cheap method for genotyping SNPs.
4.Hashimoto's thyroiditis accompanying with thyroid cancer:98 cases
Shengqian XU ; Zhiming SONG ; Guohua ZHU
Journal of Endocrine Surgery 2010;04(6):388-389,393
Objective To discuss the clinical characteristics of Hashimoto's thyroiditis accompanying with thyroid cancer. Methods Clinical data of 98 cases Hashimoto's thyroiditis accompanying with thyroid cancer were retrospectively reviewed. Results 2 cases underwent lateral thyroid lobectomy and isthmus resection. 58 cases underwent bilateral thyroid lobectomy. 35 cases underwent central lymph node dissection, with 15 cases of lymph node positive. 3 cases underwent modified radical dissection of cervical lymph nodes and all of them were proved to be lymph node positive. Metastasis rate is 25.0% and 52. 2% respectively for tumors whose diameter was less than 1 cm and greater than 1 cm. The difference has no statistic significance. Conclusions Hashimoto's thyroiditis usually accompany with thyroid papillary carcinoma and cervical lymph node metastasis can occur even if it's microcarcinoma. Lymph node metastasis rate increases with increasement of the tumor's diameter. Total thyroidectomy should be performed for Hashimoto's thyroiditis concomitant with thyroid cancer. And if necessary,lymph node dissection should be extended to the central region and lateral neck area.
5.Research advances in quantitative detection of single cell protein
Yiyun SHEN ; Xiemin QI ; Qinxin SONG ; Guohua ZHOU
Journal of China Pharmaceutical University 2015;(5):521-531
Proteins presence and differences of the expression level can clarify the physiological or pathological changes in organisms;so the quantitative detection of proteins is vital for disease mechanism research;diagnosis and prognosis evaluation.Traditional protein quantitation methods at the tissue level reflected the average protein expression in cells;but ignore the differences between individual cells.In contrast;approaches for quantitative detection at single-cell level can better reflect the differences.Recently;a number of approaches for such detec-tion have been proposed;including microfluidics;microwell-based technology;optical fiber nanobiosensor;activity-based probe technology and mass spectrometry.The principles;advantages and drawbacks of these approaches are briefly introduced in this review.
6.The expression of connective tissue growth factor and vascular endothelial growth factor in remnant rat kidney during the early process of compensatory renal growth following unilateral nephrectomy
Enfeng SONG ; Ruhan JIA ; Congxin HUANG ; Guohua DING
Journal of Chinese Physician 2001;0(06):-
Objective To study the role of connective tissue growth factor(CTGF) and vascular endothelial growth factor(VEGF) in the early stage of compensatory renal growth,and to explore the mechanisms of renal compensatory hypertrophy on rat kidney following unilateral nephrectomy.Methods The 48 Sprague Dawley rats were randomly divided into 2 groups with 24 rats in each.The rats in the operation group were underwent right nephrectomy operation and those of the control group were without any treatment.Six rats of each group were sacrificed on the 1st,4th,8th and 16th day of the experiment,respectively.The immunohistochemical method was used to investigate the expressions of CTGF and VEGF in the remnant left kidney of rats in each group.Results In the operation group,the weight of left kidney was increased obviously and the left kidney on the 16th day was obviously heavier than that on the 1st day of the experiment(P
7.Establishment of mice model that induces mucosal immunity by oral infection of Toxoplasma gondii
Hongli LIU ; Jianfeng HAN ; Guorong YIN ; Guohua SONG
Chinese Journal of Schistosomiasis Control 1989;0(03):-
Objective To establish mice model that induces mucosal immunity by orally infected tachyzoites of Toxoplasma gondii. Methods Respectively, 5?103, 5?104, 5?105, 5?106 tachyzoites of RH strain were inoculated to BALB/c mice by stomach delivery. The control group was given PBS solution. Symptoms and pathological changes of mice were observed. Secretory im-munoglobulin A (SIgA) was assayed. The lymphocytes from Peyer's patches (PP) and intraepithe-lial lymphocyte (IEL) were observed and the changes of CD4+ and CD8+ T cells assayed by SABC immunohistochemistry. Results Inoculation of 5?104 tachyzoites of Toxoplasma gondii caused symptoms and pathological changes in mice. The titre of SIgA increased in intestine, and CD4+ T subset of the mucosal inductive sites and CD8+ T subset of the mucosal effectors' sites increased. Conclusion Mucosal immunity may be induced by oral infection of 5?104 tachyzoites of T. gondii RH strain in BALB/c mice.
8.Effects of furin inhibitor on metastasis of human breast cancer MCF-7 cells
Jingli REN ; Qi SHI ; Mingzhen SUN ; Guohua SONG ; Yongchao MA
Chinese Journal of Pathophysiology 2014;(12):2267-2271
[ ABSTRACT] AIM:To investigate the mechanism underlying breast cancer metastasis and to provide theoretical da-ta for studying the pathogenesis of breast cancer onset and development.METHODS: Human breast cancer MCF-7 cells were treated with different concentrations of furin inhibitorα1-PDX for 48 h.Wound healing assay and Transwell assay were applied to detect the migration and invasion abilities of the MCF-7 cells.The expression of cell migration-associated proteins, including membrane-type 1 matrix metalloproteinase ( MT1-MMP) , vascular endothelial growth factor ( VEGF)-C and VEGF-D, was determined by Western blotting.The protein levels of MMP2 and MMP9 in the supernatant were measured by ELISA. RESULTS:Compared with control group, 200 nmol/L of furin inhibitor exerted significant inhibitory effects on the cell mi-gration (P<0.05).The expression of cell migration-associated proteins MT1-MMP, VEGF-C and VEGF-D was significantly inhibited after treated withα1-PDX ( P<0.05 ) .Significant inhibitory effects of α1-PDX on the expression of MMP9 and MMP2 (P<0.05) in the supernatant were observed.CONCLUSION:Furin inhibitor suppresses the metastasis of MCF-7 cells via down-regulating the expression of MMPs and VEGFs.
9.Establishment of Cloning and Sequencing Method for High-Resolution Human Leukocyte Antigen-B Genotype Assay
Xiaoqing XING ; Yanan CHU ; Zheng XIANG ; Qinxin SONG ; Guohua ZHOU
Chinese Journal of Analytical Chemistry 2014;(11):1574-1579
Ahigh-resolutionmethodforhumanleukocyteantigen-B(HLA-B)genotypingwasestablished based on the optimized polymerase chain reaction, cloning and sequencing technology. The exon2 and exon3 of HLA-B gene were amplified with primers based on the HLA-B gene sequence. These produced heterozygous alleles were effectively cloned into plasmid DNA based on the principle of plasmid incompatibility, and were followed by bacterial culture. Then Sanger sequencing was carried out and after analyzing the result by software ClustalX2 and IMTG/HLA database comparison, the HLA-B genotype of the samples was achieved. Seven clinical samples were detected, and the results were consistent with those of PCR-SBT genotyping method. The method was cost-effective, high-resolution and it did not require technical software. The use of universal primers simplified the cumbersome design and optimization process of specific primers.
10.Effects of fluvastatin on the tubulointerstitium in progressive diabetic kidney disease
Ping GAO ; Ruhan JIA ; Guili CHU ; Enfeng SONG ; Guohua DING ;
Chinese Journal of Geriatrics 2003;0(11):-
Objective To investigate the effects of fluvastatin on the tubulointerstitium damage in progressive diabetic kidney disease. Methods A rat model of type 2 diabetic nephropathy (DN) was developed successfully by combination of dietary induced insulin resistance and low dose STZ induced hyperglycemia after unilateral nephrectomy. Female SD rats were randomly divided into three groups: control rats, type 2 diabetic rats and type 2 diabetic rats treated with fluvastatin (2mg/kg/d). After 6 weeks, blood glucose, serum insulin, serum triglyceride and cholesterol, serum creatinine, and urinary protein were measured respectively. The protein expressions of c Jun and tansforming growth factor (TGF) ? 1 were studied by immunohistochemistry. TGF ? 1 gene expression was studied with a RT PCR technique. Results Fluvastatin at lower doses, which did not influence blood glucose and blood lipid level, significantly inhibited expression of c Jun protein(0 2536?0 0180 vs 0 5855?0 0314, P