3.Construction of eukaryotic expression vector of fusion gene CD80-IgG and its expression in Chinese hamster ovary cells
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To construct the eukaryotic expression vector CD80-IgG by fusing the cDNA encoding extracellular portion of murine CD80 to the 5'-terminus of cDNA encoding Fc fragment of murine immunoglobulin G1 and to express the fusion protein in Chinese hamster ovary (CHO) cells. METHODS: The two cDNAs was amplified by PCR respectively from plasmid pcDNA/B7 containing the full-length cDNA of murine CD80 from murine spleen cells, and cloned to the eukaryotic expression vector pcDNA3.0 by directional cloning. The resultant recombinant plasmid pcDNA/CD80-IgG was transfected into CHO cells with liposome transfection reagent. The stably expressing cells were obtained by G418 screening. Western blot, Dot ELISA, and flow cytometry were used to detect the expression of the fusion protein and its immunological activity. RESULTS: DNA sequencing verified the correction of the construction of recombinant plasmid pcDNA/CD80-IgG. The expressed fusion protein was detected in the supernatant of transfected CHO cells and the molecular weight of the protein was similar to what we expected. Its immunological activity was also established. CONCLUSION: The recombinant plasmid pcDNA/CD80-IgG was successfully constructed and it expressed the fusion protein CD80-IgG.
4.Expressions of Bile Acid Receptor FXR and TGR5 in Patients with Ulcerative Colitis
Min WEI ; Wenfang CHENG ; Jing PING ; Lu XIA
Chinese Journal of Gastroenterology 2017;22(8):465-468
Background:The incidence of ulcerative colitis (UC)in developed countries is higher than that in developing countries,which may be related with westernized lifestyle,especially high animal protein and low complex carbohydrate diet. With the increased high fat and meat intake,synthesis and secretion of bile acid in liver is also increased,which may have an impact on the occurrence of UC. Aims:To investigate the expressions of farnesoid X receptor (FXR)and G protein-coupled bile acid receptor 5 (TGR5)in patients with UC. Methods:Thirty patients with active UC from January 2013 to June 2016 at the Affiliated Jiangning Hospital of Nanjing Medical University were enrolled,and 30 healthy subjects were served as controls. Expressions of FXR and TGR5 were determined by immunohistochemistry. Results:Compared with control group,expression of FXR was significantly decreased in UC patients (4. 63 ± 2. 07 vs. 6. 91 ± 2. 62,P =0. 00),however,no significant difference in expression of TGR5 was found between the two groups (6. 70 ± 2. 90 vs. 6. 11 ± 2. 44,P = 0. 40). Expression of FXR was significantly increased in right hemicolon colitis than in left hemicolon colitis (P < 0. 05). Conclusions:There is a significant decrease in FXR in active UC patients,indicating that FXR may have some role in the pathogenesis of UC,however,TGR5 may have no obvious effect in the pathogenesis of UC.
5.Association of interleukin-6 receptor gene polymorphism with obesity
Caifeng YAN ; Hongding XIANG ; Wei CHEN ; Fan PING ; Min YANG
Chinese Journal of Endocrinology and Metabolism 2010;26(2):125-126
To examine the relationship between genetic variants in the interleukin-6 receptor gene and obesity. The result showed that the carriers of Asp358 homozygotes had higher risk in developing obesity when compared with Ala358 homozygotes (OR = 1.32,95% CI 1.07-1.68, P = 0. 041). Interleukin-6 receptor gene pulymorphism was significantly associated with obesity.
9.The significance of quantitative temperature sense thresholds in diagnosis of small fibrous sensory neuropathy in patients with type 2 diabetes.
Hou-min YIN ; Wei FENG ; Mei-ping DING
Chinese Journal of Applied Physiology 2015;31(2):150-153
OBJECTIVETo evaluate the small fiber function in patients with type 2 diabetes mellitus of the early stage by measuring the sensory threshold with the quantitative temperature testing technology.
METHODSTwenty cases of patients with type 2 diabetes with no neurological deficit (DM group) and twenty age and sex-matched healthy controls underwent the detecting of cold sensory threshold (CST), warm sensory threshold (WST), cold pain threshold (CPT), heat pain threshold (HPT) in both inside of their hands.
RESULTSThere was no significant difference in CST, WST, CPT and HPT between left and right inside of hand of the same sample among all the testers. But the four kinds of threshold showed significant difference in the right inside of hand between patients and healthy people ( P < 0.05). In addition, the CST and WST differed significantly in the left inside of hand between the patients and healthy controls while the CPT and HPT showed no significant difference in the left inside of hand between them. Patients group and control group with CST and WST on the left side of the comparison difference was statistically significant (P < 0.05).
CONCLUSIONQuantitative analysis of temperature sense threshold can not only reflect increase of the pain threshold value, also can reflect its decrease, i. e. hyperalgesia, which may help to diagnose small fibrous peripheral neuropathy recognition, especially in early diabetic peripheral neuropathy.
Case-Control Studies ; Cold Temperature ; Diabetes Mellitus, Type 2 ; physiopathology ; Diabetic Neuropathies ; diagnosis ; Hot Temperature ; Humans ; Hyperalgesia ; Pain Threshold ; Sensory Thresholds ; Thermosensing
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