2.Study on Activation of Nuclear Factor Erythroid 2-related Factor 2 Pathway with Fructus Schisandrae Extracts Medicated by Kinase Pathways Such as Extracellular Signal-regulated Kinase
Yang YU ; Qiao LAI ; Changqiang HE ; Jinlian HE ; Jie GAO
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(6):957-962
Objective To observe the influence of several kinase pathways such as extracellular signal-regulated kinase ( ERK) , and mitogen-activated protein kinase p38 ( p38MAPK) on nuclear factor erythroid 2-related factor 2 (Nrf2) pathway activated by Fructus Schisandrae extracts (FSE) . Methods HepG2 cells were treated by FSE for 24 hours after pretreatment with protein kinase inhibitors for 2 hours. The mRNA expression levels of Nrf2 and downstream target genes heme oxygenase-1 (HO-1), NAD (P) H quinine oxidoreductase 1(NQO1), P-glycoprotein ( P-gp) and multidrug resistance-associated protein 2 ( MRP2) were detected by real-time polymerase chain reaction ( RT-PCR) , and their protein expression levels and Nrf2 nuclear translocation were measured by Western blotting method. Results RT-PCR results showed that the mRNA expression levels of HO-1, NQO1, P-gp and MRP2 activated by FSE in HepG2 cells were inhibited by PD98059, SB203580 and Rottlerin, and the mRNA expression of Nrf2 was suppressed only by SB203580 and Rottlerin. Western blotting results showed that the mRNA expression levels of HO-1 and P-gp activated by SCE in HepG2 cells were inhibited by PD98059, SB203580 and Rottlerin. In addition, the protein expression of Nrf2 in HepG2 cytoplasm was increased by the above three inhibitors, and nuclear translocation of Nrf2 was inhibited by PD98059 and SB203580. Conclusion The mechanism of FSE activating Nrf2 pathway may be associated with the increase of Nrf2 nuclear translocation through the direct phosphorylation of Nrf2 induced by ERK and p38MAPK.
3.Research progress of sequencing method for animal mitochondrial genome
Tianjie LI ; Yanxiang CAO ; Hongcui ZHAO ; Yang YU ; Jie QIAO
Tianjin Medical Journal 2016;44(6):796-800
Mitochondria, the power house of cells, are important organelles in eukaryotic cells. Having their own unique and complete DNA (mtDNA) and genetic system, mitochondria play an essential role in cellular energy metabolism, intracel?lular signaling and apoptotic pathways, as well as many other biological functions, which are closely related with cellular met?abolic network. A disruption of mitochondrial genes can therefore result in mitochondrial dysfunction and human diseases, thus they have been widely used in molecular biology, development biology, genetics, forensic identification and clinical diag?nosis. Consequently, sequencing mitochondrial genome has shown great significance in mitochondrial structure and function research. In this review, research progress in mitochondrial genome sequencing method is summarized, mainly focusing on Sanger sequencing, long-PCR and next-generation sequencing. Also rolling circle amplification and indirect sequencing of mtDNA are reviewed. The ambiguities caused by numts in indirect sequencing are mentioned and resolved.
4.Expression of neuronal intermediate filament protein (NF-66) and relationship with benign and malignancy of insulinomas
Bei LIU ; Yu XIAO ; Liming ZHU ; Xinwei QIAO ; Hongding XIANG ; Jie CHEN ; Yuanjia CHEN
Chinese Journal of Pancreatology 2012;12(2):89-91
ObjectiveTo investigate whether neuronal intermediate filament protein(NF-66) could be used as a molecular marker for the determination of malignancy of insulinoma.MethodsThe expression of NF-66 protein was detected in insulinoma and pana - cancerous tissues and 3 insulinoma cell lines by immunohistochemistry staining. The relationship between the expression of NF-66 protein and the clinicopathological characteristics,survival was analyzed by univariate and multivariate statistical analysis.ResultsExpression of NF-66 was found in 102(77% ) out of 132 insulinomas and none of 98 paired control (P =4.86 × 10-31 ).NF-66 was highly expressed in 3 insulinoma cell lines.The expression of NF-66 was found in 96 (81%) out of 118 benign tumors (P =0.003),while out of 14 malignant insulinomas,6(43% ) were found to express NF-66 ( P =0.003 ).The expression of NF-66 was significantly associated with tumor size (3 cm as the cut-off point),distant metastasis (38% vs 81%,P =0.013) and distant plus lymph node metastasis (46% vs 81%,P =0.009),respectively.The expression of NF-66 was not correlated with age,gender,recurrence and overall survival.ConclusionsDown-regulation of NF-66 was significantly associated with tumor malignancy,suggesting that NF-66 could be a potentially novel molecular bionarker to distinguish malignancy from benign insulinoma.
5.Study on expression profile of mRNA in brain of pronuclear transfer mice
Tianjie LI ; Yanxiang CAO ; Xiaohu JIN ; Hongcui ZHAO ; Yang YU ; Jie QIAO
Tianjin Medical Journal 2016;44(10):1213-1216
Objective To investigate the expression profile of mRNAs in brain samples collected from pronuclear transfer (PNT) mice. Methods Female CD-1 mice were superovulated, and zygotes were collected after mating with adult male mice. Zygotes with two pronuclei were selected for pronuclear transfer manipulation, and then the reconstructed zygotes were transferred into the oviduct of pseudopregnant female mice. The infant mice obtained from pronuclear transfer were called PNT group, while the embryoes that were not performed pronuclear transfer was regarded as control group. Total RNA were extracted from brain samples of both PNT and control mice, and cDNA were labeled with fluorescent dye. Genes that were differentially expressed were identified using the Agilent mouse mRNA array. Gene ontology analysis and pathway analysis were also completed. Results Compared with control group, 392 mRNAs were expressed differentially, which showed more than 2.0 times variation and statistical significance, accounting for 1.7% of all mRNAs. Among those 366 mRNAs were up-regulated and 26 mRNAs were down-regulated. Eleven mRNAs came to 4.0 times variation in total. Gene ontology analysis indicated that differentially expressed genes were significantly enriched in alternative mRNA splicing, small GTPase mediated signal transduction, regulation of insulin receptor signaling pathway, hydrolase activity, transmembrane transporter activity and pyrophosphatase activity. Significant enriched pathway terms contained ion channel transport, fatty acid metabolism, butanoate metabolism, triacylglycerol and ketone body metabolism. Conclusion Pronuclear transfer might influence some key metabolism process in mouse brain.
6.Bacterial Community Structure in Four Different Rivers of Shanghai in Spring
Cai-Hong JIANG ; Mei-Ling ZHANG ; Yan-Jie TAO ; Na YU ; Li-Qiao CHEN ;
Microbiology 1992;0(04):-
In April of 2008 water samples were collected from four different rivers,which were Wuchao gang River,Henggang River,Chaoyang River and Caoyanghuanbang River.During the sampling the physi-cal and chemical parameters were measured.The abundance and the diversity of the bacteria of these four rivers were studied.The results showed that the population level increased in the more severely polluted river while the bacterial diversity decreased;the bacterial community structure was also affected by the dif-ferent ecological conditions of each sampling spot.The bacterial composition and abundance was closely related to the water quality in the river.
7.Effect of Duhuojisheng Decoction on primary knee osteoarthritis in macaca fascicularis
Qiao-Yu CHEN ; Xin ZHOU ; Shi-Jie FU
Chinese Journal of Tissue Engineering Research 2018;22(12):1866-1871
BACKGROUND:Meta-analysis results have shown that Duhuojisheng Decoction can improve the symptoms and relieve the pain of patients with knee osteoarthritis, but the mechanism and pathway are not very clear.OBJECTIVE:To further verify the curative effect of Duhuojisheng Decoction for primary knee osteoarthritis in macaca fascicularis. METHODS: Under natural conditions, 6 clean young macaca fascicularis (aged 3-5 years, normal group) and 6 elderly macaca fascicularis (aged more than 20 years, model group) were selected from 24 macaca fascicularis. The models of primary knee osteoarthritis were established in the model group,then aged animals were randomly divided into control and intervention(given the treatment of Duhuojisheng Decoction) groups. The number of whole blood leukocytes, C-reactive protein, erythrocyte sedimentation rate, properties of the joint fluid and expression level of tumor necrosis factor-α were compared between the control and intervention groups. RESULTS AND CONCLUSION: The aged macaca fascicularis was characterized by a typical osteoarthritis similar to human. Treatment with Duhuojisheng Decoction significantly down-regulated the expression level of tumor necrosis factor-α and significantly reduced the levels of whole blood leukocytes, and C-reactive protein. Our results suggest that the aged macaca fascicularis model of primary knee osteoarthritis is an effective animal model to simulate the occurrence and development of human primary knee osteoarthritis.Duhuojisheng decoction alleviates the progression of primary knee osteoarthritis probably by inhibiting the expression level of tumor necrosis factor-α and decreasing the levels of inflammatory factors in the articular cartilage. Additionally, the levels of whole blood leukocytes and C-reactive protein are important parameters for the prediction of curative effects.
8.The study of anterior sclera thickness and associated ocular parameters
Mei, LI ; Yu, CAI ; Ying-zi, PAN ; Rong-hua, QIAO ; Yuan, FANG ; Jie, WANG
Chinese Journal of Experimental Ophthalmology 2013;(1):81-85
Background The condition of the sclera is associated with many ocular diseases.The measurement of human scleral thickness in vivo is helpful for us to understand the features of the sclera and related diseases.Objective The present study was to measure the anterior sclera thickness(AST) in patients with senile cataract and to analyze the relationship among AST and other associated ocular parameters.Methods This study was approved by the Ethic Commission of First Hospital of Peking University.Written informed consent was obtained from each individual prior to examination.One hundred and five senile cataract patients were recruited in this study.Central corneal thickness (CCT),corneal curvature (CCV) and axial length were measured using ultrasonic pachymeter,keratometer,and A-scan unit,respectively.The AST was measured at 2 mm posterior to the scleral spur in the temporal meridian using ultrasound biomicroscope (UBM).The differences of CCT,CCV,ocular axial length and AST between bilateral eyes and the different sexes were compared by the Paired test and independent sample t test.The correlations among various parameters were assessed by the Pearson linear correlation analysis.The differences of CCT and AST among different axial length groups were evaluated by one-way ANOVA.Results No significant differences were found in the CCT,CCV,axial length and AST between bilateral eyes (t =0.584,P =0.561 ; t =1.161,P =0.248 ; t =0.140,P =0.889 ; t =0.342,P =0.773).Temporal AST at 2 mm posterior to the sclera spur was (0.589 ±0.051)mm in the right eyes.An insignificant decline in CCT was found in the male group compared with the female group (right eyes:t =0.469,P =0.641 ; left eyes:t =0.465,P =0.643).However,compared with the female group,the increase of axial length,reduction of the mean CCV value and enhancement of the mean AST were observed(right eyes:all P<0.01 ;left eyes:all P<0.01).CCV showed a negative correlation with ocular axial length (r =-0.50,P<0.01),but no significant correlation was found among age,CCT,ocular axial length and AST(P>0.05).No remarkable differences were found in CCT and AST among the various axial length groups (CCT:F =0.998,P=0.372;AST:F=1.919,P=0.383).Conclusions In senile cataract patients,correlation is not found between AST and CCT;the increase of axial length is not associated with the thinning of the eyeball wall to a certain extent.Differences exist in some ocular parameters between different sexes.
9.Measurement of anterior scleral thickness and its correlation with central corneal thickness and axial length in different types of glaucoma patients
Mei, LI ; Rong-hua, QIAO ; Ying-zi, PAN ; Yu, CAI ; Yuan, FANG ; Jie, WANG
Chinese Journal of Experimental Ophthalmology 2013;(3):249-253
Background A close relation between sclera thickness and glaucoma has been determined.Clinical features vary in different types of glaucoma patients,which hints that the scleral thickness might be distinct among these patients.Objective The aim of this study was to investigate the differences of central corneal thickness(CCT),anterior scleral thickness(AST) and axial length in glaucomatous patients.Methods This study was approved by the Ethic Commission of First Hospital of Peking University.Written informed consent was obtained from each individual prior to the examination.A retrospective descriptive study was designed.One hundred and sixty consecutive patients were recruited from March,2009 to November,2010 in First Hospital of Peking University,including 35 eyes with primary angle closure glaucoma(PACG) (35 cases),34 eyes with primary open-angle glaucoma POAG) (34 cases),37 eyes with normal tension glaucoma(NTG) (37 cases)and 17 eyes of ocular hypertension OHT) (17 cases).Thirty-seven eyes of 37 subjects with incipient cataract served as the control group.CCT and ocular axial length were measured with ultrasonic pachymeter and A-scan unit,respectively,and AST at the temporal quadrant 2 mm posterior to the scleral spur was measured by ultrasonic biomicroscopy (UBM).The measuring parameters among different groups were compared by analysis of covariance,and the correlations of AST,CCT with ocular axial length were analyzed using Pearson linear correlation and linear regression.The differences and correlation of CCT,AST and AL among five groups were analyzed.Results The CCT values in PACG group,POAG group,NTG group,OHT group and control group were (535.54 ± 5.20),(550.47 ± 5.28),(521.61 ± 5.07),(575.75 ± 7.76) and (535.06± 5.06) μm,respectively,showing a significant difference among them (F =9.560,P =0.000),and the CCT value of OHT group was increased in comparison with POAG group,PACG group,NTG group and control group(all P =0.000).The CCT of the POAG group was thicker than that in the PACG group,NTG group and control group(P=0.046,0.000,0.040).No significant difference was found in CCT among NTG group,PACG group and control group(P=0.950,0.060).The AST values of PACG group,POAG group,NTG group,OHT group and control group were(0.593±0.050),(0.600±0.050),(0.592-±0.060),(0.610-±0.060) and(0.604±0.060) mm,respectively,showing a insignificant difference among them (F =0.700,P =0.590).The axial length in the patients with PACG was shorter than that of the POAG group,NTG group,OHT group and control group (all P =0.000).The Pearson correlation analysis showed significantly positive correlation between CCT and AST in POAG group and NTG group(r=0.445,P=0.008;r=0.400,P=0.014).Conclusions This study confirms that there is dissimilarity in CCT but not in AST among different types of glaucomatous patients.The changes of CCT and AST are consistent in POAG and NTG patients.
10.Changes of glucagon and somatostatin levels during oral glucose tolerance test in patients with type 2 diabetes mellitus
Limin MA ; Fangzhen XIA ; Hui ZHU ; Yi CHEN ; Jiao YU ; Ying LI ; Yanxiang LI ; Huixin ZHANG ; Jie QIAO ; Yingli LU
Chinese Journal of Endocrinology and Metabolism 2011;27(9):749-751
The fasting and 2 h levels of glucagon, somatostatin ( SS), and C-peptide during 75 g oral glucose tolerance test in 60 patients with type 2 diabetes and 34 normal subjects were determined. Compared with control group, the fasting levels of glucagon and SS and 2 h levels of SS after glucose loading significantly decreased,while the fasting and 2 h levels of C-peptide increased in diabetes group. The 2 h levels of these hormones were significantly higher than the fasting levels in two groups. Compared with control group, the increased folds of glucagon ( 1.40±0.48 vs 1.20±0. 30, P<0. 05 ) and SS( 2.79±2. 17 vs 1.14±0. 22, P<0. 01 ) levels after glucose loading were higher and that of C-peptide level ( 3.58 ±3. 10 vs 8. 33 ± 6. 99, P<0. 01 ) was lower in diabetes group. The levels of fasting glucagon were positively correlated with that of fasting SS in two groups( both P<0. 01 ). These results suggest that disturbance exists in hormones from α and δ cells besides the dysfunction of β cells in patients with type 2 diabetes.