1.Effects of sinomenine on cell activation and intracellular Th1 type cytokines expression of T lymphocytes
Xiaojuan LI ; Peixun WANG ; Liang LIU ; Guangxing CHEN ; Jingxian ZHAO ; Yanying ZENG ; Jipan CHEN
Chinese Journal of Immunology 1986;0(04):-
Objective:To study deeply on the effects of sinomenine on Th1 type cytokines expression of T lymphocytes.Methods:The authors isolated and cultured the mesenteric lymph nodes of mice in vitro.The lymphocytes were stimulated with PDB and inomycin after added sinomenine in different concentrations for 1 hour.After 4 hours stimulation,cells were collected and stained intracellular cytokines with the dying regents,then the expression of Th1 type cytokine IFN-?,pro-inflammatory cytokine TNF-? of T lymphocytes were analyzed by flow cytometry.And also observed the effects of drugs on the expression of CD69,an early marker of T cell activation.Results:1 000 ?mol/L(329.4 ?g/ml)sinomenine can downregulate CD69 expression of T lymphocytes while 200 ?mol/L sinomenie show no effect on it.200,1 000,2 000 ?mol/L sinomenine can obviously reduce the expression of intracellular cytokines such as IFN-?,TNF-? in dose-dependent manner.Conclusion:One of the immunological mechanisms of sinomenine to treat rheumatoid arthritis may due to the inhibitory effects on abnormal T lymphoctytes activation.The inhibitory effects of sinomenine may mainly be relative to the inhibitory effects on Ca 2+ -dependant signal pathway of T lymphocytes activation but not PKC pathway.
2.Research of the best way to the double-labeling immunofluorescence staining
Jipan YAN ; Yong ZHENG ; Hao LIU ; Rui LI ; Ning ZHANG ; Weigang CHEN
Journal of Chinese Physician 2012;(z1):4-6
Objective Detailed descriptions of the double-labeling immunofluorescence staining for fluorescence microscopy provides an ideal sample.Methods Rat liver frozen sections were used fixative were 95% alcohol,95% formaldehyde and acetone,frozen sections,with anti-CSE,Ki-67 polyclonal antibody and incubated with FITC,Cy3 fluorescence-labeled secondary fluorescently labeled secondary antibody staining,observed under a fluorescence microscope.Results Acetone fixed group visible in the proliferative phase (S phase) cells showed a red fluorescent nucleus,cytoplasmic green fluorescence.Conclusion The impact of double labeling immunofluorescence the effect of sample links and many factors,including the two most important factors are 2 and coordination with the primary antibody and select the appropriate fixative.
3.Clinical characteristics and CYP17A1 gene mutation analysis in patients with 17α-hydroxylase/17, 20-lyase deficiency and testicular tumor
Binbin HAN ; Ruizhi ZHENG ; Yidan XIE ; Yiqi CHEN ; Jipan NIU ; Yun ZHANG
Chinese Journal of Internal Medicine 2021;60(9):827-830
The 17α-hydroxylase/17, 20-lyase deficiency (17-OHD) is a rare disease. The clinical characteristics and gene mutation of 2 late-diagnosed 17-OHD patients with testicular tumor admitted to our hospital from March 2018 to February 2019 were analyzed retrospectively. The two 17-OHD patients were female (46, XY). Laparoscopic abdominal exploration found undeveloped testicles in grey-yellow or grey-red in the groin and iliac fossa. The testicles were removed and showed malignancy in pathology study. Sequencing of the CYP17A1 gene identified c.1247G>A/c.1427T>C and c.985_987delTACinsAA/c.1306G>A complex heterozygous mutations. Taking together, the possibility of 17-OHD should be considered in patients with hypertension, hypokalemia, adrenal adenomatoid hyperplasia together with 46, XY gonadal dysplasia, so as to make early diagnosis and treatment, and avoid dysplastic testicular turning to malignancy.