1.Effects of Silencing G6PD Expression on The Growth and Apoptosis in Human Skin Melanoma
Yuechun ZHU ; Huiru LU ; Danyi LI ; Shufen TONG
Progress in Biochemistry and Biophysics 2006;0(03):-
Glucose-6-phosphate dehydrogenase (G6PD) derives from the expression of the house-keeping gene G6PD. Recent studies have indicated that G6PD is related to tumor genesis, growth, clinical phenotype, therapy, and prognosis. To elucidate the relationship between G6PD and cancer, three siRNA sequences and one negative control sequence were designed based on the 3' noncoding region of the human G6PD gene. Two complementary single-strand DNA (sense and antisense) were designed and synthesized based on siRNA sequences. The DNA fragments were annealed and ligated to the GFP expression vector pRNAT-U6.2/Lenti. One siRNA with higher interference efficiency than the other two was found after siRNA plasmid transfecting human skin A375 melanoma cells. After lentivirus particle packaging and virus production, the A375 cells were infected, and the single cell clone was acquired and cultured to establish the stable cell strain. Western blotting showed that the endogenous G6PD in the stable A375 cell strain was 0.257 ? 0.074, which was 11.17% of G6PD expression (2.301 ? 0.286) in wild type A375 cells. The final siRNA interference efficiency in this stable cell strain was 88.83%. The G6PD activity of A375-G6PD?驻 was 21.53% of A375-WT. Further study showed that A375-G6PD△ doubling generation time prolonged, and its proliferation was greatly inhibited and the cloning efficiency lowered 25%(P
2.THE EFFECT OF MACROPHAGE-CONDITIONED MEDIUM ON THE GROWTH OF PRIMARY MICE MYOBLASTS
Danyi SHU ; Junmin TANG ; Yan TANG ; Feng LI
Acta Anatomica Sinica 1955;0(03):-
Objective To investigate the effect of macrophage-conditioned medium on the growth of mouse myoblasts. Methods After a 3-day intraperitoneal injection with a mixture of muscle homogenate and starch,macrophages were obtained from mouse peritoneal cavity and cultured for 48 hours in serum-free DMEM/F12.Then the cultured medium was harvested.Myoblasts were obtained from newborn mice skeletal muscle and grown in DMEM/F12 supplied with 10% FBS for 36-48 hours,and then exposed to 0.5% FBS medium with or without macrophage-conditioned medium.Cells were harvested at 72 hours later,their growth was observed with Wright staining,LDH cytochemistry and immunocytochemistry. Results Macrophage-conditioned medium can maintain the normal morphology and the activity of myoblasts grown in low concentration of serum and promote their proliferation.There was a significant difference between average absorbance and integral absorbance of LDH positive cells of the two groups(P
3.The inhibition and possible mechanism of resveratrol on hepatocellular carcinoma cell line Bel-7402 both in vitro and vivo
Danyi ZHAO ; Chaoxia DAI ; Jun CHEN ; Dan LI ; Wentao GAO
Chinese Journal of Biochemical Pharmaceutics 2014;37(7):6-8,13
Objective To explore the inhibition effect and the possible mechanism of resveratrol on human hepatocellular carcinoma cell line Bel-7402 both in vitro and vivo.Methods Four Res drugs in the experiment group,the final concentrations were 12.5,25,50,100μmol/L,the control group at the same time set not containing Res drugs,MTT assay was used to measure the inhibition of resveratrol on Bel-7402.The expression of Bcl-2 was detected by RT-PCR and Western Blot.The levels of IL-2,IL-6,IL-12 and TNF-αwere detected by ELISA.Results Resveratrol inhibited Bel-7402 cell proliferation in dose and time manner,and influenced the expression of Bcl-2 mRNA and protein.At the same time,resveratrol inhibited the growth of tumor and improved the levels of IL-2,IL-6,IL-12 and TNF-α.Conclusion Resveratrol could inhibit Bel-7402 cell proliferation both in vitro and vivo, the possible mechanism may be that resveratrol could low down the expression of Bcl-2 and improve the levels of IL-2,IL-6,IL-12 and TNF-α.
4.A new 3, 4-seco-lanostane triterpenoid from a marine-derived fungus Ascotricha sp. ZJ-M-5.
Leirui XIE ; Danyi LI ; Peile WANG ; Huiming HUA ; Xin WU ; Zhanlin LI
Acta Pharmaceutica Sinica 2013;48(1):89-93
To study the secondary metabolites of a marine-derived fungus Ascotricha sp. ZJ-M-5, several chromatographic methods including macroporous resin, silica gel, ODS and Sephadex LH-20 were used to isolate the compounds, and their structures were elucidated on the basis of physicochemical properties and spectroscopic methods. Ten compounds were obtained and identified as ascotrichic acid B (1), (3R)-6-hydroxymellein (2), beta-carboline (3), (22E, 24R)-ergosta-7, 22-diene-3beta, 5alpha, 6beta-triol (4), (22E, 24R)-ergosta-7, 22-diene-3beta, 5alpha, 6beta, 9alpha-tetraol (5), cyclo (Leu-Pro) (6), cyclo (Ile-Leu) (7), cyclo (Pro-Val) (8), cyclo (Pro-Gly) (9), and cyclo (Hpro-Ala) (10). Among them, compound 1 is a new 3, 4-seco-lanostane triterpenoid which has been isolated from the filamentous fungi for the first time, and compounds 2-10 are firstly isolated from Ascotricha genus.
5.Validation of Microbial Limit Test for Compound Clindamycin Hydrochloride Gel
Jinlei LI ; Jiayi WANG ; Danyi CHI ; Qian ZHU ; Yi WANG ; Wenhuan FU
China Pharmacy 2007;0(25):-
OBJECTIVE:To establish microbial limit test for Compound clindamycin hydrochloride gel and to carry out the validation of three methods.METHODS:According to the appendices volumeⅡof Chinese Pharmacopeia(edition 2005),microbial limit test which contained routine test,medium dilution method and membrane-filter method was adopted to validate Compound clindamycin hydrochloride gel.Recoveries of three methods were compared to make sure the suitable validation method.RESULTS:In routine test,recovery of trial bacteria was lower than 70%and controlled bacteria couldn't be found out.In medium di-lution method,recovery of trial bacteria was higher than 70%and still lower than the recovery of diluents.In membrane-filter method,recovery of trial bacteria was higher than 90%,and controlled bacteria grew well.Membrane-filter method could eliminate inhibition effect of Compound clindamycin hydrochloride gel on bacteria.CONCLUSION:Membrane-filter method is effective and practical for microbial limit test of Compound clindamycin hydrochloride gel.
6.Expression of Far Upstream Element Binding Protein 1 in Gastric Cancer and Its Effect on Proliferation and Invasion of Gastric Cancer Cells
Dan LI ; Baojun HUANG ; Danyi ZHAO
Journal of China Medical University 2017;46(7):587-590
Objective To investigate the expression of far upstream element binding protein 1 (FUBP1) gene in gastric cancer,and analyze its effect on proliferation and invasion of gastricc cancer cells.Methods The expression of FUBP1 in gastric cancer tissues was detected by real-time PCR.The relationship between FUBP1 expression and clinic pathological factors of gastricc cancer was analyzed by one-way ANOVA.The silence vector for FUBP1 was constructed and transfected into gastricc cancer SGC7901 cells,and the transfected effects were then detected by real-time PCR.The influence of FUBP1 silence on cell proliferation and invasion in SGC7901 cells were detected by CCK8 method and Transwell assay.Results FUBP1 expression was increased markedly in gastricc cancer tissues.Following the depressing of differentiation and the increase of invasion depth and metastatic lymph nodes,the FUBP1 expression was up-regulated in primary gastric carcinoma significantly.The pS-FUBP1 vector could silence the FUBP1 expression in SGC7901 cells.FUBP1 silence inhibited SGC7901 cell proliferation amd invasion.Conclusion FUBP1 was high-expressed in gastricc cancer,and related with genesis and progress of gastric cancer,silence of FUBP1 expression can inhibit the cell proliferation and invasion in gastric cancer cells.
7.Effect of Q-switched ruby laser combined with retinol topical serum on periorbital hyperpigmentation
Danyi HUANG ; Ting LIU ; Li YANG ; Yiming LIN ; Han MA
Chinese Journal of Medical Aesthetics and Cosmetology 2023;29(5):397-400
Objective:To assess the effectiveness and safety of retinol topical serum combined with Q-switch ruby laser in the treatment of periorbital hyperpigmentation (PH).Methods:From October 2021 to December 2021, 30 patients with PH were treated in the Department of Dermatology, the Fifth Affiliated Hospital of Sun Yat-sen University, including 7 males and 23 females, aged 24-33 years, with an average age of 28.9 years. The patients were divided into a combination therapy group of 20 cases and a laser therapy group of 10 cases. In the combined treatment group, 20 subjects received a Q-switch ruby laser once a month, three times in total, and the retinol topical serum were used after the treatment for three months; the others were treated solely with laser.Results:In the combined treatment group, there was significant difference in visual analogue scale (VAS) before and after the use of retinol topical serum ( t=3.21, P=0.005) and in Sadick tear trough rating scale before and after treatment ( t=2.67, P=0.015). After therapy, 4 cases obtained good improvement and 13 cases obtained some improvement in the overall evaluation of the investigator; 17 patients had good improvement and some improvement. The overall satisfaction scores of 17 subjects were very satisfied. In the laser therapy group, there was no statistically significant difference in VAS before and after treatment in the simple laser treatment group ( t=0.29, P=0.782), and there was no statistically significant difference in the Sadick tear trough rating scale ( t=1.56, P=0.154). Among them, in the overall evaluation of the researchers, 1 case (10%) achieved good improvement, 3 cases (30%) achieved some improvement. The overall satisfaction score of 4 subjects was very satisfactory. The combination therapy group had the highest effective rate and satisfaction, and the difference between the two groups was statistically significant (χ 2=14.089, P=0.002). Conclusions:The application of retinol topical serum combined with Q-switched ruby laser in periorbital hyperpigmentation is safe and effective, and is worthy of clinical promotion.
8.Study on the role of teachers in the multidisciplinary integrated PBL teaching in pediatrics with the investigation of student satisfaction
Yanhong LUO ; Danyi PENG ; Yanan LI ; Yongming WANG ; Gaohui ZHU ; Daoqi WU ; Ziyu HUA ; Enmei LIU ; Jianfeng HE
Chinese Journal of Medical Education Research 2018;17(3):248-254
Objective To discuss the role of teachers in the integrated PBL teaching of pediatrics in the light of the investigation of students after PBL teaching.Method The multidisciplinary integrated PBL course established by the Department of Pediatrics of Chongqing Medical University has been implemented for more than two years in the teaching of the five year program in the Department of clinical medicine (pediatric direction).From September 28,2016 to October 25th,the researchers conducted a survey of 26 undergraduate students in the college who received PBL teaching at the professional probation stage.After the end of each PBL class,the teaching evaluation questionnaire recommended by Fudan University and Qian Ruizhe was distributed through the network teaching platform of Chongqing Medical University.The questionnaire was retrieved by students without a name,and the survey was carried out 3 times.The data of the questionnaire were summarized with Excel 2007.Result 77 questionnaires were issued and 77 were recovered,with a recovery rate of 100%.In the multidisciplinary integrated PBL class,94.8%-97.4% students gave high score evaluation,and all the evaluation of teachers reached "excellent" level.In response to open-ended questions,students believed that teachers in teaching could inspire students to think positively,encourage students to express different views,create a mutually trusted environment for students,and cultivate students' interpersonal communication,communication and cooperation ability.Conclusion In pediatric multidisciplinary integrated PBL teaching,teachers should teach students correct learning methods,judge each student's knowledge and thinking level,and individualize teaching for each student's problems and characteristics.
9.Microbiome research outlook: past, present, and future.
Yunyun GAO ; Danyi LI ; Yong-Xin LIU
Protein & Cell 2023;14(10):709-712
10.Effect of high glucose peritoneal dialysis solution on PGC-1α expression and mitochondria related oxidative injury in human peritoneal mesothelial cells.
Xuejing ZHU ; Feng WEN ; Danyi YANG ; Jing LIU ; Shuguang YUAN ; Jun LI ; Hong LIU ; Xiangqing XU ; Lin SUN ; Fuyou LIU
Journal of Central South University(Medical Sciences) 2013;38(11):1085-1091
OBJECTIVE:
To investigate the mechanism of mitochondrial oxidative injury induced by high glucose peritoneal dialysis solution (PDS) and the protective effect of peroxisome proliferator activated receptor gamma coactivator 1-alpha (PGC-1α) in the mitochondria of human peritoneal mesothelial cells (HPMC) in the high glucose ambience.
METHODS:
HPMC was cultured in a PDS containing 1.5%, 2.5% and 4.25% glucose for 24 hours. Western blot analysis was used to detect PGC-1α expression. MitoSOX? Red staining, respiratory chain complexes and antioxidant enzyme activities were determined.
RESULTS:
The activities of respiratory chain complex III and antioxidant enzymes decreased significantly in a concentration- and time-dependent manner, along with the increased production of mitochondrial reactive oxygen species (ROS) and cellular apoptosis. In addition, protein expression of PGC-1α was also decreased in the high glucose PDS ambience.
CONCLUSION
High glucose PDS might inhibit PGC-1α expression, resulting in the inhibition of mitochondrial function and increase of mitochondrial ROS and cellular apoptosis.
Apoptosis
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Dialysis Solutions
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adverse effects
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Epithelial Cells
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pathology
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Glucose
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adverse effects
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Humans
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Mitochondria
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metabolism
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pathology
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Oxidative Stress
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Peritoneal Dialysis
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Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
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Reactive Oxygen Species
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Transcription Factors
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metabolism