1.Research progress in mitigation of endoplasmic reticulum stress by glucagon like peptide-1
Qianqian WANG ; Yuanyuan XIAO ; Li WEI
Chinese Journal of Endocrinology and Metabolism 2015;(2):188-191
Glucagon-like peptide-1(GLP-1) is an incretin stimulated by food mainly produced and secreted from L-cells in terminal ileum, colon and rectum. It can be combined with GLP-1 receptors, and then plays a series of biological effects. In recent years, studies have shown that GLP-1 participates in the occurrence and development of many diseases.
2.Effects of triptolide on the expression of androgen receptor in human prostate LNCaP cells and its mechanism of action.
Bide LIU ; Qianqian FENG ; Xiao GU ; Dan LU ; Wei LI
Acta Pharmaceutica Sinica 2015;50(10):1246-51
To study the regulation of androgen receptor (AR) expression in human prostate cancer LNCaP cells by triptolide (TP) and the possible mechanism, by using qRT-PCR and Western blot, the AR mRNA and protein levels in TP treated LNCaP cells were detected, and the AR protein level in TP and NF-κB inhibitor treated LNCaP cells was also detected; a series of pGL3-AR promoter reporter gene vectors were built using restriction-free cloning method, and the vectors were employed to investigate the effects of TP on the transcriptional activity of AR promoter in LNCaP cells; the upstream proteins which may play regulatory roles were detected using western blot assay. After treated LNCaP cells with TP for 48 h, AR mRNA and protein expressions decreased with increasing TP concentration. The expression of AR target gene PART1 and prostate specific antigen (PSA) was also downregulated by TP treatment; a series of pGL3-AR promoter reporter vectors were constructed and validated by sequencing and luciferase activity; the results of dual luciferase reporter assay showed that TP downregulated AR at the transcriptional level; PI3K/AKT/NF-κB pathway which is associated with AR promoter activity was drowregulated by TP. In conclusion, our results demonstrated that the transcriptional activity of AR in LNCAP cells was downregulated by TP, and PI3K/AKT/NF-κB pathway may be involved in the regulation mechanism.
3.piggyBac transposon system and its applications in the genemanipulation in mammals
Qianqian WANG ; Hui ZHANG ; Liang XIAO ; Liming ZHANG
Chinese Journal of Biochemical Pharmaceutics 2017;37(1):302-305
DNA transposon is a kind of factor that is able to translocate gene in its genome, thus offering an efficient method for permanently modifying the genome of mammals. The piggyBac (PB) transposon system has been proven effective in mammalian genomic engineering, including cancer gene discovery, animal transgenesis, in vivo gene delivery as well as in vitro genetic modification like induced pluripotent stem cells. In addition, piggyBac has many desirable features, such as seamless excision of transposons from the genomic DNA and the potential to target integration events to desired DNA sequences. Therefore, the piggyBac translocation system is an ideal genetic tool in the construction of animal models and gene therapy, and we can anticipate that the piggyBac transposon system will play a more and more important role in biomedical research.
4.Influence of RNA interference on MMP-9 gene expression in THP-1 cell line
Li YU ; Yunzhi LING ; Xiao XIAO ; Angxuan HAN ; Ke PENG ; Pengchen XU ; Qianqian GAO
Chinese Journal of Immunology 2014;(8):1069-1073
To study the inhibitory effect of RNA interference (RNAi) on MMP-9 gene expression in THP-1 cell line.To investigate the application of RNAi on the therapy of leukemia.Methods:Small interfering RNA ( siRNA) for MMP-9 gene was designed and transfected into THP-1 cells.MMP-9 mRNA expression was assessed by RT-PCR, and MMP-9 protein expression was tested by Western blot.MTT and trypan blue staining were used to observe the effect on the proliferation of THP-1 cells after RNAi.The changes in cell morphology were observed under the microscope.Results:The expressions of MMP-9 mRNA and protein were inhibited in THP-1 MMP-9 siRNA-transfected cells ,significantly lower than those of control cells.The results of MTT and trypan blue staining in-dicated that the proliferation ability of THP-1 cells obviously decreased after siRNA-transfected 48h and 72h.The growth of cells was in-hibited and the cells survival rate was significantly lower than that of control group ( P<0.05 ).The cells of control groups grew semi-quote wall under inverted microscope.The outline of cells was clear and the shape was uniform.The cells grew vigorously.While the growth of cells in siRNA group was inhibited.The morphology of siRNA group cells changed obviously by the Wright staining.Most cells expressed changes of apoptosis.Conclusion: siRNA for MMP-9 gene can not only reduce the expressions of MMP-9 mRNA and protein,but also inhibit the proliferation and induce apoptosis of THP-1 cells.
5.The application of MRI in sporadic inclusion-body myositis
Qianqian YUAN ; Jiangxi XIAO ; Ying ZHU ; Lingchao MENG ; Yun YUAN ; Xiaoying WANG
Chinese Journal of Radiology 2016;50(5):376-379
Objective To investigate the characteristic MRI features of sporadic inclusion?body myositis(sIBM). Methods Clinical and MR imaging data of 6 patients with sIBM diagnosed by muscle biopsy from May 2013 to November 2014 were retrospectively analyzed. All patients showed insidious onset of lower limb muscle weakness and diagnosed as sIBM by muscle biopsies. All patients were evaluated by the score of the severity of fatty infiltration, inflammation and atrophy in MRI. Results All patients were observed fatty infiltration with different degrees. The fatty infiltration in thighs was characterized in a decreasing order of frequency:gluteus maximus (6 cases), vastuslateralis (6 cases), vastusintermedius (6 cases), vastusmedialis (6 cases), sartorius (5 cases), adductor magnus (5 cases), rectus femoris (4 cases), semi?membranosus (4 cases), semi?tendinosus (4 cases), biceps femoris (4 cases), gracilis (3 cases), adductor longus(2 cases).The fatty infiltration in thighs was characterized in a decreasing order of severity:vastuslateralis (3.2 points), vastusintermedius (3.2 points), vastusmedialis (3.0 points), adductor magnus (3.0 points), gluteus maximus (2.7 points), bicepsfemoris (2.2 points), semi?membranosus (2.1 points), semi?tendinosus (2.1 points), rectus femoris (1.5 points), sartorius (1.3 points), gracilis (0.8 points), adductor longus (0.7 points). All patients showed the features of distal distribution andsymmetry. Inflammation was observed in 3 patients. 1 patient only involved the vastuslateralis, the other 2 patients were observed muscle inflammation with different degrees in 12 muscles. Atrophy was observed in 5 patients. The atrophy in thighs was characterized in a decreasing order of frequency:vastuslateralis (5 cases), vastusintermedius (5 cases), vastusmedialis (4 cases), adductor magnus (4 cases), semi?membranosus (2 cases), rectus femoris (1 cases), sartorius (1 cases) and gluteus maximus;there was no atrophy in adductor longus, gracilis,semi?tendinosus, biceps femoris. Conclusion The MRI characteristic manifestations of sIBM is fatty infiltration and atrophy in the distal portion, particularly involving the vastuslateralis, vastusintermedius, vastusmedialis and adductor magnus.
7.Effects of hyperosmolality on expression of urea transporter A2 and aquaporin 2 in mouse medullary collecting duct cells.
Wenmin, JIN ; Xi, YAO ; Taoxia, WANG ; Qianqian, JI ; Yongxia, LI ; Xiao, YANG ; Lijun, YAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(1):59-64
In this study, the effects of hyperosmolality on the expression of urea transporter A2 (UTA2) and aquaporin 2 (AQP2) were investigated in transfected immortalized mouse medullary collecting duct (mIMCD3) cell line. AQP2-GFP-pCMV6 and UTA2-GFP-pCMV6 plasmids were stably transfected into mIMCD3 cells respectively. Transfected mIMCD3 and control cells were cultured in different hypertonic media, which were made by NaCl alone, urea alone, or an equiosmolar mixture of NaCl and urea. The mRNA and protein expression of AQP2 was elevated by the stimulation of NaCl alone, urea alone and NaCl plus urea in AQP2-mIMCD3 cells; whereas NaCl alone and NaCl plus urea rather than urea alone increased the mRNA and protein expression of UTA2 in UTA2-mIMCD3 cells, and all the expression presented an osmolality-dependent manner. Moreover, the mRNA and protein expression of UTA2 rather than AQP2 was found to be synergistically up-regulated by a combination of NaCl and urea in mIMCD3 cells. It is concluded that NaCl and urea synergistically induce the expression of UTA2 rather than AQP2 in mIMCD3 cells, and hyperosmolality probably mediates the expression of AQP2 and UTA2 through different mechanisms.
8.Protective effects of Dextran-40 on fatal jellyfish stings
Hui ZHANG ; Qianqian WANG ; Liang XIAO ; Chao WANG ; Bo WANG ; Beilei WANG ; Liming ZHANG
Chinese Journal of Biochemical Pharmaceutics 2017;37(5):1-6
Objective To investigate the protective effects of Dextran-40 on fatal jellyfish stings at whole animal and cellular levels.MethodsFirstly, using the fatal jellyfish envenomation syndrome (acute jellyfish envenomation syndrome, AJES and delayed jellyfish envenomation syndrome, DJES) models established earlier by ourselves, we surveyed the effects of Dextran-40 on the survival rate of AJES mice, on the circulatory function of AJES rats and on the serum biochemical indexes of DJES rats.In addition, the protective effects of Dextran-40 on cardiomyocytes against the damage induced by tentacle extract (TE) from the jellyfish Cyanea capillata were studied at the cellular level by laser scanning confocal microscopy.ResultsAt the whole animal level, Dextran-40 at 0.8g/kg significantly improved the survival rate of AJES mice, and at 0.6g/kg greatly inhibited the drop of blood pressure of AJES rats.For the DJES rats, Dextran-40 at 0.6g/kg had a significant protective effect on the liver function indexes (ALT, AST, A/G) and myocardial enzymes (CK, LDH).At the cellular level, Dextran-40 5μg/mL greatly inhibited the TE-induced increase in intracellular Ca2+ content and the death of cardiomyocytes.ConclusionThe protective effects of Dextran-40 on fatal jellyfish stings may be related to its ability to stabilize blood pressure or block the TE-induced pore formation in the cardiomyocytes.Considering that the clinical safety of Dextran-40, we strongly recommend it as a first-aid medicine for jellyfish stings.
9.Simultaneous determination of five active components in resina draconis and its extract by HPLC.
Yun LI ; Wei XIAO ; Jianping QIN ; Ying GUO ; Qianqian ZHU
China Journal of Chinese Materia Medica 2012;37(7):929-933
OBJECTIVETo establish an HPLC method for simultaneous determination of contents of loureirin A, loureirin B, 7,4'-dihydroxy flavone, pterostilbene and resveratrol in resina draconis and its extracts.
METHODKromasil 100-5C18 column (4.6 mm x 250 mm, 5 microm) was used with the mobile phase of acetonitrile-1% glacial acetic acid at a flow rate of 1.0 mL x min(-1) and the column temperature at 40 degrees C. The detective wave length of loureirin A and B was detected at 278 nm, and 7,4'-dihydroxy flavone, pterostilbene and resveratrol was at 319 nm.
RESULTAll the five active components reached the resolved peaks within 40 min, indicating a good linearity (r > or = 0.999 7). The average recoveries of loureirin A, loureirin B, 7,4'-dihydroxy flavone, pterostilbene and resveratrol in resina draconis were 102.9%, 96.81%, 97.29%, 100.7% and 103.7%, with RSDs of 0.23%, 1.5%, 0.42%, 0.58% and 0.34%, respectively. The average recoveries of loureirin A, loureirin B, 7,4'-dihydroxy flavone, pterostilbene and resveratrol in extract of resina draconis were 102. 2% , 96. 93%, 97. 90% , 102.0% and 103.3%, with RSDs of 1.7%, 0.91%, 1.4%, 1.5% and 1.2%, respectively.
CONCLUSIONThe method is so easy, accurate, highly repeatable and stable that it provides good reference for the quality control of resina draconis and its extracts.
Chalcones ; analysis ; Chromatography, High Pressure Liquid ; methods ; Dracaena ; chemistry ; Flavones ; analysis ; Resins, Plant ; analysis ; Stilbenes ; analysis
10.mRNA expression of silent information regulator 6 among population in Bama county of Guangxi
Meiyu JIANG ; Qianqian HUANG ; Liwei SHI ; Tianlong XIA ; Xue LIANG ; Min HE ; Zhiyong ZHANG ; Jian QIN ; Xiao WEI
The Journal of Practical Medicine 2016;32(4):548-551
Objective To analyze mRNA expression of silent information regulator 6 (SIRT6) gene in the blood of population with family history of longevity in Bama county of Guangxi and to explore its association with SIRT6 gene polymorphism and its protein Methods One hundred and thirty-seven people (aged 30 ~ 106, 70 males, 67 females, 6.57% Han nationality, 93.43% Zhuang and Yao nationalities) with family history of longevity (long-lived family history group), and 91 people (aged 22~89, 51 males, 40 females, all Zhuang and Yao nationalities) without family history of longevity were recruited in the study (non-long-lived family history group). Real-time fluorescence quantitative RT-PCR was used to detect mRNA expression of SIRT6 gene in two groups. Results SIRT6 mRNA expression of total and femals in long-lived family history group were higher than those in non-long-lived family history group (P<0.05). mRNA expression of GG and CG genotype of SIRT6 (rs350846) and G allele carriers in long-lived family history group were all higher than those in non-long-lived family history group and the difference was statistically significant (P<0.05). SIRT6 mRNA expression was not associated with serum SIRT6 protein in long-lived family history group (P>0.05). Conclusion The expression of mRNA in SIRT6 gene may be associated with familial aggregation of longevity in Bama County of Guangxi and high expression of mRNA of SIRT6 in G allele carriers contributes to longevity.