1.Mechanism of alternative splicing and its application in diagnosis and treatment of leukemia
Jing LIU ; Jing ZHANG ; Bo HUANG ; Xiaozhong WANG
Chinese Journal of Laboratory Medicine 2015;38(11):730-732
Alternative splicing is a process refering a pre-mRNA transforms to different mature mRNAs by different splicing sites combination, and then the mRNAs translate to various proteins.This process is regulated by a variety of cis acting elements and trans acting factors.Recent studies have shown that aberrant alternative splicing is prevalent in leukemia patients, and is closely associated with leukemic occurrence, development and chemotherapy resistance.This shows us that aberrant altemative splicing may be helpful to leukemia diagnosis and treatment.
2.Exploration and Thinking of Designable Experiment in Hematological Examination Teaching
Xiaozhong WANG ; Jing LI ; Yongmei ZHOU ; Lingling ZHANG ; Yanfei RAO
Chinese Journal of Medical Education Research 2006;0(11):-
Designable experiment refers to such an experiment that students apply their knowledge and experiment skills to design and prepare experimental materials,instruments,means and procedures based on specific purpose and requirements,and finish the formal experiment by themselves.Through designable experiment,students' integration capability such as applying knowledge,manual operation and handling problem are cultured.We explore setting up designable experiment in the course of clinical hematological examination,and evaluate the effectiveness of teaching.
3.Establishment and identification of HEK293 cell lines with stable and high expression of NTCP
Ya CHEN ; Jing LI ; Wenzheng ZHANG ; Min LUO ; Xiaozhong FU ; Ting LIU
Chinese Pharmacological Bulletin 2016;32(12):1767-1771,1772
Abstrate:Aim To construct HEK293 cell line with stable and high expression of sodium taurocholate cotransporting polypeptide (NTCP ) efficiently and rapidly.Method Vector expressing EGFP-NTCP fusion protein was constructed and verified by DNA sequencing.The pEGFP-NTCP expression vector was transfected into HEK293 cells by FuGENE 6 transfection reagent. The transfected cells with high expression of green fluorescent protein were selected using fluorescence microscope for screening of G418 for 14 days to obtain cell lines stably and highly expressing NTCP.NTCP expression was detected by RT-PCR,qRT-PCR, Western-blot and the uptake experiment of taurocholic acid.Re-sult RT-PCR,qRT-PCR,Western-blot and the uptake experi-ment revealed that compared to the control cells,the expression of NTCP was significantly positive (P<0.01)in stable trans-fected cells showing green fluorescence (P<0.05 ).Conclusion The HEK293 cell line with stable and high expression of NTCP has been established efficiently and rapidly,which provides a cellular model for the study of the mechanism of the uptake of bile acid derivatives.
4.Comparison of hPepT1 transfected MDCK cells to hPepT1 transfected HeLa cells
Min LUO ; Xiaozhong FU ; Tao XIAO ; Wenzheng ZHANG ; Jing LI ; Ya CHEN ; Ting LIU
Chinese Pharmacological Bulletin 2017;33(2):280-284
Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.
5.The mechanism of delayed rectifier potassium channel regulated by cyclooxygenase -2 in gastric cancer cells
Xiaodong SHAO ; Kaichun WU ; Xiaozhong GUO ; Manjiang XIE ; Jing ZHANG ; Daiming FAN
Chinese Journal of Digestion 2009;29(1):46-49
Objective To investigate the currents impact on delayed rectifier potassium (HERG)regulated by cyclooxygenase (COX)-2 in gastric cancer cells and its mechnism. Methods ① The HERG mRNA, protein and current in gastric cancer cells transfected with or without COX-2 antisense vector were measured by RT-PCR, Western blot and patch-clamp, respectively. ② cAMP concentration in gastric cancer cells transfected with or without COX-2 antisense vector was measured by ELISA. ③ The mutant HERG, which was absence of cAMP-binding domain, was constructed by PCR and transfected into gastric cancer cells. ④ The impact of COX-2 inhibitor and proglandin (PG) E2 on HERG current in gastric cancer cells transfected with or without mutant HERG was investigated by patch clamp. ⑤ The effects of agonist and antagonist of cAMP and inhibitor of protein kinase (PK) A on HERG current in gastric cancer cells transfected with or without HERG mutant were observed by patch clamp. Results ① The expression of HERG mRNA and protein in gastric cancer cells transfected with COX-2 antisense vector were not altered, but the amplitude of HERG current was diminished (P<0.05). ② The cAMP concentration in gastric cancer cells transfected with COX-2 antisense vector was lower than that in parental gastric cancer cells (P<0.05). ③ COX-2 inhibitor and PGE2 had influence on the HERG currents in gastric cancer cells. COX-2 inhibitor reduced and PGE2 enhanced the amplitude of HERG current in gastric cancer cells. However, neither COX-2 inhibitor nor PGE2 showed any negative or positive effects on currents of mutant HERG. ④ cAMP agonist enhanced the amplitude of HERG current and cAMP antagonist reduced the amplitude in gastric cancer cells. Neither agonist nor antagonist had effect on currents of mutant HERG. ⑤ PKA inhibitor did not influence the HERG current of parental gastric cancer cells and gastric cancer cells transfected with mutant. Conclusions COX-2 regulates HERG current through its catalytic product of PGE2, which binds with its receptor on the gastric cancer cells and alters cAMP level in gastric cancer cells, cAMP interacts with HERG protein by binding with cAMP-binding domain of HERG protein and exerts impact on HERG current. PKA does not participate in this process.
6.Effects of transplanting bone marrow stromal cells on axonal and glial scarring after spinal cord injury
Jing WANG ; Xiaozhong ZHOU ; Chao LI ; Hongtao SUN ; Weijian CHEN ; Guitao LI
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(1):25-30
Objective To investigate the effects of bone marrow stromal cell (BMSC) transplantation on axonal and glial scarring after spinal cord injury (SCI).Methods Thirty New Zealand white rabbits were randomly assigned to a sham operation group (group A),a saline treatment group (group B) or a BMSC treatment group (group C).Group A served as controls,in which the canal was opened without damage to the spinal cord.In groups C and B SCI models were established with aneurysm clips and the rabbits of groups C and B were then given injections of BMSCs and saline solution respectively via the intra-intercostal artery at 1 week post injury.At 1 day,1 week,2 weeks and 4 weeks post injury,Basso Beattie-Bresnahan (BBB) scores were assessed to evaluate the recovery of locomotor function in the hind limbs.Spinal cord samples were harvested for HE and Nissl staining,and immunohistochemistry and image analysis were used to detect any changes in neurofilament (NF200) and glial fibrillary acidic protein (GFAP) in the injured spinal cords.Results The average BBB scores of group A were significantly higher those that of groups B and C at each time point,and those of group C were significantly better than those of group B at the 2nd and 4th week post injury.At the 4th week post injury,HE staining showed there was no glial scarring or cavities in group A,but that there was glial cellular proliferation,glial scarring and cavity formation at the injury site in groups B and C.In group C all were obviously less than in group B.Nissl staining indicated there were more typical neurons in group A,while there were a larger number of ruptured neurons,more degradation,and irregular remaining neurons in groups B and C.These abnormalities were again significantly more prevalent in group C.Immunohistochemical examination showed significant increases in NF200 positive neurons and GFAP in groups B and C compared with group A.The number of NF200 positive neurons was significantly higher in group C than in group B,but the GFAP positive area was significantly smaller in group C than in group B.Conclusion BMSC transplantation via the intercostal arteries can effectively improve axonal regeneration,attenuate glial cellular proliferation and reduce glial scar formation,promoting functional recovery after SCI,at least in rabbits.
7.Significance of alternative splicing in drug resistance of chronic myeloid leukemia
Tingyu QIN ; Jing LIU ; Xiaozhong WANG
Chinese Journal of Clinical Oncology 2019;46(16):861-864
Alternative splicing (AS) is a process by which the transcriptome diversity, and thereby the proteome diversity, is augment-ed by splicing or joining together different parts of the pre-mRNA in eukaryotic cells . AS at different splice sites is regulated by multi-ple cis-acting elements and trans-acting factors. Chronic myeloid leukemia (CML) is a clonal myeloproliferative disease in which there is a translocation between the long arms of chromosome 9 and chromosome 22, represented as t(9;22) (q34;q11). This translocation results in the formation of a BCR-ABL fusion gene. Hence it is not surprising that resistance to tyrosine kinase inhibitors, which inhibit BCR-ABL activity, has become a critical problem in the clinical treatment of CML. Using second generation high-throughput sequencing technology, it has been found that AS abnormalities are closely related to the occurrence, progression, drug resistance, and immune escape of CML. This paper reviews the research related to AS and CML resistance and investigates the potential causes of CML resis-tance. Drug resistance mechanisms and potential therapeutic targets are also reviewed.
8.Design, synthesis and anti-oxidative evaluation of L-amino acid prodrugs of scutellarein.
Xiaozhong FU ; Wei ZHANG ; Yonglin WANG ; Yanyu LAN ; Aimin WANG ; Wen ZHOU ; Yong HUANG ; Jing LI ; Fengjing XING ; Ying LIU
Acta Pharmaceutica Sinica 2011;46(5):548-55
To design and synthesize a series of novel scutellarein 4'-L-amino acid prodrugs with more potent anti-oxidative activity and improved physicochemical properties. Scutellarein was used as lead compound, according to successful experience of improving bioavailability of oral administration drugs by active transport mechanism, principle of hybridization was used to introducing L-amino acid structural fragments at 4'-position of scutellarein to design and synthesize target scutellarein 4'-L-amino acid prodrugs. The synthetic compounds were tested on their physicochemical properties and in vitro anti-oxidative activity against H202 induced oxidative damage in PC12 cells. Five compounds were found to have more potent anti-oxidative activity than positive control VE. Moreover the physicochemical properties of synthesized compounds were evaluated, and the results revealed that L-amino acid ether derivatives are more stable (t1/2 9-92 h) than their corresponding ester derivatives (t1/2 0.5 h). Water solubility of scutellarein 4'-L-amino acid ester and ether derivatives were 1 796-4 100 microg.mL-1 and 27.7-81.1 microg.mL-1 respectively, in comparison with scutellarin, the solubility of compounds 18, 19 and 22, 24-27 increased about 120-280 fold and 2-6 fold respectively. All these results suggested that L-amino acid prodrug strategy has significant potential in scutellarein prodrug design.
9.A research of the pertinence between of serum uric acid level and coronary artery CTA SYNTAX score
Yongbin LI ; Jing JIA ; Ning WANG ; Jiayi DU ; Xiaozhong SHI ; Yuanfei QU ; Chunhai LUO
Journal of Practical Radiology 2017;33(11):1744-1746
Objective To explore the correlation between serum uric acid level and coronary artery SYNTAX score of coronary heart disease.Methods A total of 69 patients of coronary heart disease were enrolled according to SYNTAX score.The patients were divided into the low risk group (27 cases),medium risk group (23 cases) and high risk group (19 cases).The differences of serum uric acid concentrations among the three groups were compared by ANOVA.Spearman rank correlation analysis was used to analyze the correlation between serum uric acid concentrations and coronary artery SYNTAX scores.Results ANOVA analysis showed that the differences of serum uric acid concentrations among the groups were statistical significant (F=4.74,P<0.05).The Spearman correlation analysis showed that serum uric acid concentrations were positively correlated with coronary SYNTAX score (r =0.58,P <0.05).Conclusion Serum uric acid level and severity of coronary artery disease are positively correlative.
10.Association between liver fibrosis and gut microbiota based on "harmonizing liver and spleen"
Yunxiao LIU ; Jing DOU ; Xiaozhong WANG
Journal of Clinical Hepatology 2023;39(2):278-283
Liver fibrosis is the leading cause of the morbidity and mortality of liver diseases worldwide and has become one of the key issues in the research on chronic liver diseases among domestic and foreign scholars. An increasing amount of evidence has shown that gut microbiota dysbiosis plays a crucial role in the development and progression of liver fibrosis, and improvement of gut microbiota dysbiosis has become a new target for anti-fibrotic treatment. At present, traditional Chinese medicine has attracted much attention in improving gut microbiota. Studies have shown that "harmonizing liver and spleen" plays an important role in reducing fibrosis degree and delaying the progression of liver fibrosis. Therefore, based on the theory of "harmonizing liver and spleen", the achievements in liver-gut axis and gut microbiota by modern molecular biological techniques can provide a theoretical basis for the treatment of liver fibrosis from "harmonizing liver and spleen" in clinical practice, as well as new directions and ideas for anti-fibrotic treatment with traditional Chinese medicine.