1.Reproduction of a rat model of metabolic syndrome to study the related gene expression
Tingbing CAO ; Zhencheng YAN ; Chengyi SHEN
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
Objective To reproduce a rat model of metabolic syndrome (MS) to analyze the variations of related gene expression. Methods 30 male rats aged 8w were randomly divided into two groups, the rats in NC group (control) were fed with normal diet (10% fat and 0.5% salt ), and those in metabolic syndrome (MS) group with high fat diet (49% fat and 2% salt). The body weight, blood pressure, fasting blood glucose (FBG), blood lipid and fasting insulin level were serially measured. Such feedings were continued for 24 weeks, and then the intraperitoneal glucose tolerance test and hyperinsuline-euglycemic gomphosis test were performed, and carotid arterial pressure and visceral fat were measured. RT-PCR was used to detect the genes related to energy consumption, glucose-lipid metabolism in white adipose, brown adipose and muscle tissue. Results Compared with NC group, all the variables were increased significantly, such as body weight, visceral fat weight, blood pressure, serum levels of TG and FFA. A marked insulin-resistance and decreased glucose tolerance were found in MS group. Hyperinsuline-euglycemic gomphosis test revealed that the mRNA expression of 23 genes related to glucose-lipid and energy metabolism changed significantly in white adipose, brown adipose and muscle tissue in MS group as compared with NC group. Conclusion Prolonged high fat plus high salt diet may cause the clinical features of MS in rats. The changes in various genes may be involved in the mechanisms involved in the pathogenesis.
2.Analysis on Quality of Rheum Palmatum L. from Gansu Province Based on Multicriteria Method
Guodi LU ; Chengyi LI ; Huizhen WANG ; Fude YANG ; Mingwei WANG ; Junzhong SHEN
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(10):57-63
Objective To compare the quality of Rheum palmatum L. in different areas and ages in Gansu Province by comparing five components in Rheum palmatum L. through fingerprints and QAMS (quantitative analysis of multi-components) methods; To determine the most suitable growing areas and the best development areas in Gansu Province. Methods The water content, total ashes, and water-soluble extract content in 15 batches of Rheum palmatum L. from different areas and ages in Gansu Province were determined according to the approaches listed in the Chinese Pharmacopeia. The contents of five index components were determined through HPLC and the quality of 15 batches of Rheum palmatum L were evaluated by fingerprint and QAMS method. Results The results from 15 batches of Rheum palmatum L. showed that the water content was less than 15%, total ashes less than 10%, and water-soluble extract content less than 25%, conforming to Pharmacopoeia. There were 23 common peaks in the fingerprints of 15 batches of Rheum palmatum L.. Five of them were identified as aloe-emodin, rhein, emodin, chrysophanol and physcion, which showed good linear relationship in the range of 0.0122–0.7344 μg (r=0.9999), 0.00714–5.7120 μg (r=0.9999), 0.0088–0.7040 μg (r=1.0000), 0.1224–3.6720 μg (r=0.9999) and 0.0148–5.9200 μg (r=0.9997), respectively. There was no significant difference between the calculated value and the measured value by using the relative correction factor (RCF), and the reproducibility of RCF was good. The quality of Rheum palmatum L. from different areas and ages in Gansu Province significantly differed (P<0.05). Two year old Rheum palmatum L. from Pingxiang village of Li county and Rushu village of Tanchang county had the best quality, and that of three years old was better than two years old from Tanchang county. Conclusion The established fingerprints and QAMS method is accurate, feasible, and can be used for the quality comparison of Rheum palmatum L. from different areas and ages in Gansu Province. Li County and Tanchang County areas in Gansu Province can be used as suitable planting areas and development industry. Therefore, it is recommended to select more than two yeas old Rheum palmatum L. from Tanchang County and Li County in Gansu Province for medicinal application.
3.Protective effect of Schisandrin B on myocardial cells with hypoxia/reoxygenation-induced inj ury in neonatal rats and its mechanism
Wei LIU ; Chengyi ZHANG ; Nan SHEN ; Hong JIN ; Rui YUAN ; Xiaoming WAN ; Ling QI
Journal of Jilin University(Medicine Edition) 2014;(5):977-980
Objective To investigate the influence of Schizandrain B(Sch B)preconditioning in myocardial cells of neonatal rats with hypoxia/reoxygenation (H/R)inj ury, and to explore its possible mechanism.Methods The cultured myocardial cells were divided into control group, model group and 10, 50, 250 mg · L-1 SchB preconditioning groups.All the cells in various groups except control group were cultured in 2 mmol·L-1 Na2 S2 O4 for 2 h,then cultured in normal medium for 18 h to induce myocardial H/R injury.The morphological changes of myocardial cells in various groups were observed under inverted microscope. The survival rates of the myocardial cells in each group were examined by MTT. The activities of lactic dehydrogenase (LDH ), creatine (CK ), superoxide ismutase(SOD),and the (MDA)levels in the cells in various groupswere examined by detection kits. Results Compared with control group,the cells in model group were retracted,arrest or float,the survival rate was decreased significantly (P<0.01),the activities of LDH,CK and MDA level were increased (P<0.01),and the SOD activity was decreased (P<0.01);compared with model group,the cells in Sch B preconditioning groups remained beating,retraction was light,the cell survival rates were significantly increased (P<0.01),the activities of LDH,CK and MDA levels were decreased (P<0.01),and the SOD activities were increased (P<0.05 or P<0.01).Conclusion Sch B has protective effect on myocardial cells with H/R injury in the neonatal rats,which may be associated with anti-oxidative damage.
4.Expression and clinical significance of galectin 3 in bladder cancer tissues and its effect on malignant biological behavior of T24 cells
LUO Le ; LI Qiujiang ; LONG Xingxia ; SHEN Chengyi ; ZHENG Zaiyong ; WEN Aiping
Chinese Journal of Cancer Biotherapy 2018;25(5):503-508
[Abstract] Objective: To investigate the expression of galectin-3 in bladder cancer tissues and its correlation with clinicopathological characteristics of bladder cancer patients, as well as to explore its effect on the proliferation, invasion and apoptosis of bladder cancer T24 cells. Methods:Atotal of 104 cases of pathologically confirmed bladder cancer tissues and the corresponding adjacent tissues were collected from the patients treated at the Affiliated Hospital of North Sichuan Medical College from May 2014 to June 2016. Immunohistochemistry staining was used to determine the galectin-3 protein expression in both cancer and adjacent tissues, and the correlations between galectin-3 expression and clinical pathological features were analyzed. siRNA-Gal3 and siRNA-Control were transfected into T24 cell, respectively. The expression of galectin-3 protein was detected by Western blotting, the proliferation of cells was detected by MTT assay; the invasion of cells was detected by Transwell assay; and the cell apoptosis was determined by Flow cytometry. Results: The positive rate of galectin-3 in bladder cancer tissues was significantly higher than that in adjacent tissues (73.1% vs 9.6%, P<0.05). The expression of galectin-3 in bladder cancer was correlated with histological grade, depth of invasion, lymph node metastasis and TNM stage (all P <0.05), but not with sex and age (P>0.05). The expression of galectin-3 was down-regulated significantly by siRNAGal3 (P<0.05). After interference with galectin-3, the proliferation and invasion of T24 cells was significantly decreased (all P<0.05) but the apoptosis was significantly increased (P<0.05). Conclusion: Galectin-3 is over-expressed in bladder cancer and is closely related to the clinicopathological features of bladder cancer patients. Interference of galectin-3 protein expression can inhibit proliferation and invasion and promote cell apoptosis of bladder cancer T24 cells.
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5.Annotation of the mobilomes of nine teleost species.
Bo GAO ; Dan SHEN ; Cai CHEN ; Saisai WANG ; Kunlun YANG ; Wei CHEN ; Wei WANG ; Li ZHANG ; Chengyi SONG
Chinese Journal of Biotechnology 2018;34(1):122-131
In this study, the mobilomes of nine teleost species were annotated by bioinformatics methods. Both of the mobilome size and constitute displayed a significant difference in 9 species of teleost fishes. The species of mobilome content ranking from high to low were zebrafish, medaka, tilapia, coelacanth, platyfish, cod, stickleback, tetradon and fugu. Mobilome content and genome size were positively correlated. The DNA transposons displayed higher diversity and larger variation in teleost (0.50% to 38.37%), was a major determinant of differences in teleost mobilomes, and hAT and Tc/Mariner superfamily were the major DNA transposons in teleost. RNA transposons also exhibited high diversity in teleost, LINE transposons accounted for 0.53% to 5.75% teleost genomic sequences, and 14 superfamilies were detected. L1, L2, RTE and Rex retrotransposons obtained significant amplification. While LTR displayed low amplification in most teleost with less than 2% of genome coverages, except in zebrafish and stickleback, where LTR reachs 5.58% and 2.51% of genome coverages respectively. And 6 LTR superfamilies (Copia, DIRS, ERV, Gypsy, Ngaro and Pao) were detected in the teleost, and Gypsy exhibits obvious amplication among them. While the SINE represents the weakest ampification types in teleost, only within zebrafish and coelacanth, it represents 3.28% and 5.64% of genome coverages, in the other 7 teleost, it occupies less than 1% of genomes, and tRNA, 5S and MIR families of SINE have a certain degree of amplification in some teleosts. This study shows that the teleost display high diversity and large variation of mobilome, there is a strong correlation with the size variations of genomes and mobilome contents in teleost, mobilome is an important factor in determining the teleost genome size.
6.Enhancer trapping nearby rps26 gene in zebrafish mediated by the Tol2 transposon and it's annotation.
Yatong SANG ; Dan SHEN ; Wei CHEN ; Shuheng CHAN ; Hao GU ; Bo GAO ; Chengyi SONG
Chinese Journal of Biotechnology 2018;34(3):449-458
With the completion of large-scale genome sequencing of human beings and other organisms, understanding the expression of control elements on the genome has become an important research task in the post-genome era. The enhancer trapping technology is an effective method for identifying enhancer elements in the genome and understanding its mechanism for gene expression regulation. In this study, we selected the stable enhancer trapping line TK4 (head and trunk specific GFP expression), which is generated with the mediation of Tol2 transposon system, and analyzed the trapped enhancers with the techniques of Splinkerette PCR (sp-PCR), in situ hybridization and comparative genomics. We crossed F1 individuals of TK4 line with wild-type zebrafish, collected fertilized eggs, and then detected the expression pattern of green fluorescent protein reporter gene by fluorescence microscopy at six different developmental stages, 6 hpf (hour post fertilization), 24 hpf, 48 hpf, 3 dpf (day post fertilization), 4 dpf and 5 dpf . The zebrafish genome flank sequence near the insertion site of Tol2 transposon was cloned by sp-PCR, and the results revealed that the insertion located at the position 27749253 of chromosome 23, and the transgene inserted reversely inside the intron 1 of rps26 gene. Within the 100 kb region of the insertion site, totally, seven genes including arf3a, wnt10b, wnt1, rps26, IKZF4, dnajc22 and lmbr1l were identified. Comparative genomic analysis by VISTA program revealed that there were two potential enhancer elements in the downstream of rps26 gene, which were conserved non-coding sequence (CNS) 1 and CNS2. The results of in situ hybridization showed that two transcripts of rps26 gene were maternal expression, the expression of rps26-201 in zygote was earlier than that of rps26-001, and the GFP signal of TK4 line zebrafish was not detectable before 6hpf, the expression patterns of rps26 and GFP at the late stages display similarity, and also represent differences, which suggested that the expression of rps26 and GFP may be controlled by the same enhancer, and also by the different enhancer, and two potential enhancers (CNS1 and CNS2) may play a differential regulation roles on the spatial and temporal expression of nearby genes (including rps26). In this study, we successfully obtained two potential enhancers near rps26 gene for the first time, which laid a foundation for further study of the regulation mechanism between these two enhancers and nearby genes in the genome, and the combination technique used in this study also provides a reference for enhancer analysis.