1.The using of tongue diagnosis in fatty liver patients' health management
Shenghua WANG ; Rong LIANG ; Zhaoping WANG ; Fangling LI ; Jiping REN ; Rumei KANG
Chinese Journal of Health Management 2008;2(1):39-41
Objective To find the syndrome tendency of fatty liver people and then providing basis for TCM integrated into fatty liver patients'Health Management by analyzing the characteristics of fatty liver people's tongue and combining them with the investigating results of symptoms.Methods Using 1∶1control study methods,examined 368 cases(184 cases were patients with fatty liver),observed their tongues as well as their symptoms,and recorded results.Results x2 test showed fat tongue,thin and small tongue,teeth-marked tongue,thick fur,greasy fur,fond of cool drink,hate feat,diarrhea after eating cold food,more sweat,lethargy,heavy four limbs,more flatus were significantly increase in fatty liver people(x27·580,11.740,23.700,8.666,10.793,P>0.05).Conclusions The symptoms of patients with fatty liver disease were the reflection of heat and(or)damp-heat.Found the generally pathology rules of patients with fatty liver disease,we could be more reasonable and effective prevention and treatment.
2.Removal of anti-phospholipase A2 receptor autoantibodies in primary membranous nephropathy by recombinant phospholipase A2 receptor tandem epitope immunosorbent
Kexin WANG ; Shuxin LIU ; Fangling JI ; Jiahang QIAN ; Jun REN ; Lulu HAN ; Lingyun JIA
Chinese Journal of Nephrology 2023;39(11):840-845
Objective:To investigate the role of recombinant phospholipase A2 receptor (PLA2R) tandem dominant epitopes (PLA2RTD) in the removal of anti-PLA2R autoantibodies (anti-PLA2R) from primary membranous nephropathy (PMN).Methods:The recombinant protein PLA2RTD (cysteine-rich domain, C-type lectin like domain 1 and C-type lectin like domain 7) was expressed in bacmid-insect cell expression system. Circular dichroism was used to determine the secondary structure of PLA2RTD. Enzyme-linked immunosorbent assay and immunofluorescence were used to determine the biological activity of PLA2RTD. Epoxy activation method was used to couple the recombinant PLA2RTD and agarose gel CL-6B microspheres for preparing specific immune adsorbent of anti-PLA2R.Results:The study achieved the expression of PLA2RTD in the first time from the bacmid-insect cell system, demonstrating the good immunogenicity and high binding specificity of PLA2RTD. A single in vitro adsorption of PLA2RTD could averagely eliminate 76.66% of anti-PLA2R [(6.66±0.30) RU/ml vs. (28.54±2.10) RU/ml], the changes of IgG, IgA, albumin, β2 microglobulin, interleukin 6, and tumor necrosis factor α were all less than 4% after completion of adsorption, and the second or third repeated use of PLA2RTD could maintain the adsorption efficiency of about 65%. Conclusion:PLA2RTD-based specific immunosorbent can effectively remove anti-PLA2R in plasma, which provides a new way to specifically remove PMN-related autoantibodies.
4.Extraction of exosome by gel electrophoresis microfluidic chip and determination of miRNA-21 in exosome of human plasma.
Dan LUO ; Fengying RAN ; Lun WU ; Juan ZHANG ; Fangling REN ; Jingjian LIU ; Binqiang ZHANG ; Qinhua CHEN
Chinese Journal of Biotechnology 2021;37(2):663-672
We developed a high-efficiency microfluidic chip for extracting exosomes from human plasma. We collected peripheral blood from normal human, designed and fabricated a microfluidic chip based on nanoporous membrane and agarose gel electrophoresis to isolate exosomes. The extracted exosomes were characterized by transmission electron microscopy, nanosight and Western blotting, the morphology, concentration and particle size of exosomes were identified and analyzed. Meanwhile, we used ultracentrifugation and microfluidic chip to isolate exosomes separately. The particle size and concentration of the exosomes extracted by two methods were compared and analyzed, and their respective extraction efficiency was discussed. Finally, the expression level of miRNA-21 in exosomes was analyzed by RT-PCR. The microfluidic chip isolated (in 1 hour) high-purity exosomes with size ranging from 30-200 nm directly from human plasma, allowing downstream exosomal miRNA analysis. By comparing with ultracentrifugation, the isolation yield of microfluidic chip was 3.80 times higher than ultracentrifugation when the volume of plasma sample less than 100 μL. The optimized parameters for exosome isolation by gel electrophoresis microfluidic chip were: voltage: 100 V; concentration of agarose gel: 1.0%; flow rate of injection pump: 0.1 mL/h. The gel electrophoresis microfluidic chips could rapidly and efficiently isolate the exosomes, showing great potential in the research of exosomes and cancer biomarkers.
Exosomes
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Humans
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MicroRNAs/genetics*
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Microfluidics
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Plasma
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Ultracentrifugation