1.Effect of pterostilbene on the growth, apoptosis and autophagy of a human papillomavirus type 16-immortalized cervical epithelial cell line H8
Quan CHEN ; Yi TANG ; Huaping LI ; Jiaoquan CHEN ; Liqian PENG ; Ridong YANG ; Huiyan DENG ; Zhenjie LI ; Huilan ZHU
Chinese Journal of Dermatology 2021;54(10):861-868
Objective:To investigate the effect of pterostilbene on the growth, apoptosis and autophagy of a human papillomavirus type 16 (HPV-16) -immortalized cervical epithelial cell line H8.Methods:H8 cells were treated with pterostilbene at different concentrations of 0 (control group) , 25, 50, 75, 100 μmol/L for 24 and 48 hours. Cell counting kit-8 (CCK8) assay was performed to evaluate the cellular proliferative activity, flow cytometry was conducted to detect apoptosis and cell cycle, monodansylcadaverine (MDC) staining and fluorescence microscopy were performed to detect autophagy, and Western blot analysis was conducted to determine the expression of the cell cycle-related protein cyclinD1, apoptosis-related proteins caspase-3 and caspase-9, autophagy-related proteins Beclin1, microtubule-associated protein 1 light chain 3 (LC3) -Ⅱ/Ⅰ, ATG5 and P62, as well as HPV oncoproteins E6 and E7. Statistical analysis was carried out by using one-way analysis of variance, repeated measures analysis of variance and least significant difference- t test. Results:After 48-hour treatment with pterostilbene at different concentrations of 0, 25, 50, 75, 100 μmol/L, the relative cellular proliferation rate significantly differed among the groups (100.00% ± 1.56%, 99.02% ± 4.97%, 93.59% ± 2.01%, 81.28% ± 4.90%, 69.17% ± 7.56%, respectively; F = 77.22, P < 0.05) , and gradually decreased along with the increase in the concentration of pterostilbene; compared with the control group, the pterostilbene groups all showed significantly decreased cellular proliferation rate (all P < 0.05) . After 24-hour treatment with pterostilbene, the proportions of H8 cells at G1, G2 and S phases significantly differed among the above groups ( F = 7 845.00, 51.14, 266.50, respectively, all P < 0.05) ; compared with the control group, the pterostilbene groups showed significantly increased proportions of H8 cells at G1 and G2 phases (all P < 0.05) , but significantly decreased proportions of H8 cells at S phase ( P < 0.05) . After 48-hour treatment with pterostilbene, the apoptosis rate was significantly higher in the 25-, 50-, 75- and 100-μmol/L pterostilbene groups (14.66% ± 0.22%, 13.50% ± 0.49%, 14.56% ± 0.19%, 15.30% ± 0.76%, respectively) than in the control group (11.58% ± 0.50%, all P < 0.05) . After 24-hour treatment with pterostilbene, MDC staining showed only a small number of H8 cells with bright dot-like fluorescence in the control group, but increased number of autophagosome-positive H8 cells with bright dot-like fluorescence in the pterostilbene groups. Western blot analysis revealed that there were significant differences in the protein expression of cyclin D1, caspase-3, caspase-9, Beclin1, LC3-Ⅱ/Ⅰ, ATG5, P62, E6 and E7 among the control and pterostilbene groups after 24- and 48-hour treatment with pterostilbene (all P < 0.05) . The treatment with pterostilbene could down-regulate the expression of cyclin D1, E6 and E7, and up-regulate the expression of caspase-3, caspase-9, Beclin1, LC3-Ⅱ/Ⅰ, ATG5 and P62, with significant differences between the control group and most pterostilbene groups in expression of the above proteins (all P < 0.05) . Conclusion:Pterostilbene can inhibit the proliferation of H8 cells, promote their apoptosis and autophagy, and down-regulate the expression of oncogenes E6 and E7.
2.Mid- and long-term results of endovascular aneurysm repair for abdominal aortic aneurysms: a single center′s experience for 10 years
Rui WANG ; Chen YAO ; Jinsong WANG ; Zuojun HU ; Mian WANG ; Zilun LI ; Ridong WU ; Siwen WANG ; Junjie NING ; Yuansen QIN ; Yi SHI ; Jin CUI ; Guangqi CHANG
Chinese Journal of Surgery 2020;58(11):841-846
Objective:To examine the mid- and long-term outcomes of endovascular aneurysm repair (EVAR).Methods:This was a retrospective cohort study of 540 patients with abdominal aortic aneurysm who received EVAR at Department of Vascular Surgery, The First Affiliated Hospital, Sun Yat-sen University from January 2009 to December 2018. There were 503 males and 37 females, aged of (69±8) years (range: 44 to 87 years). Clinical data including concomitant disease, aneurysm size and surgical data were collected and patients were followed up after operation. The cumulative survival rate was assessed using the Kaplan-Meier estimator and multivariate Cox regression was used to analyze the independent prognosis factors.Results:The technical success rate was 91.1% (492/540). The perioperative mortality rate was 1.3% (7/540) and the follow-up rate was 91.7% (489/533). The median follow-up time was 45(63) months (range: 1 to 133 months). The all-cause mortality rate was 21.3% (104/489) and the aneurysm-related mortality rate was 6.3% (31/489) during follow-up period. The overall cumulative survival rate of 1-, 3-, 5- and 10-year were 95.1%, 84.0%, 69.5% and 38.6%, respectively, while freedom from aneurysm-related death were 98.4%, 93.3%, 88.4% and 84.4%. During the follow-up period, the complications rate was 9.0% (44/489), and the re-intervention rate was 4.9% (24/489). Cox regression analysis showed that elder age ( HR=2.15, 95 %CI: 1.41 to 3.26, P<0.01), preoperative aneurysm rupture ( HR=2.72, 95 %CI: 1.78 to 4.15, P<0.01) and short neck aneurysm ( HR=1.97, 95 %CI: 1.07 to 3.61, P=0.029) were independent prognosis factors for long-term survival after EVAR. Connclusion:EVAR has low perioperative mortality, high technical success rate, and satisfactory mid-and long-term outcomes.
3.Mid- and long-term results of endovascular aneurysm repair for abdominal aortic aneurysms: a single center′s experience for 10 years
Rui WANG ; Chen YAO ; Jinsong WANG ; Zuojun HU ; Mian WANG ; Zilun LI ; Ridong WU ; Siwen WANG ; Junjie NING ; Yuansen QIN ; Yi SHI ; Jin CUI ; Guangqi CHANG
Chinese Journal of Surgery 2020;58(11):841-846
Objective:To examine the mid- and long-term outcomes of endovascular aneurysm repair (EVAR).Methods:This was a retrospective cohort study of 540 patients with abdominal aortic aneurysm who received EVAR at Department of Vascular Surgery, The First Affiliated Hospital, Sun Yat-sen University from January 2009 to December 2018. There were 503 males and 37 females, aged of (69±8) years (range: 44 to 87 years). Clinical data including concomitant disease, aneurysm size and surgical data were collected and patients were followed up after operation. The cumulative survival rate was assessed using the Kaplan-Meier estimator and multivariate Cox regression was used to analyze the independent prognosis factors.Results:The technical success rate was 91.1% (492/540). The perioperative mortality rate was 1.3% (7/540) and the follow-up rate was 91.7% (489/533). The median follow-up time was 45(63) months (range: 1 to 133 months). The all-cause mortality rate was 21.3% (104/489) and the aneurysm-related mortality rate was 6.3% (31/489) during follow-up period. The overall cumulative survival rate of 1-, 3-, 5- and 10-year were 95.1%, 84.0%, 69.5% and 38.6%, respectively, while freedom from aneurysm-related death were 98.4%, 93.3%, 88.4% and 84.4%. During the follow-up period, the complications rate was 9.0% (44/489), and the re-intervention rate was 4.9% (24/489). Cox regression analysis showed that elder age ( HR=2.15, 95 %CI: 1.41 to 3.26, P<0.01), preoperative aneurysm rupture ( HR=2.72, 95 %CI: 1.78 to 4.15, P<0.01) and short neck aneurysm ( HR=1.97, 95 %CI: 1.07 to 3.61, P=0.029) were independent prognosis factors for long-term survival after EVAR. Connclusion:EVAR has low perioperative mortality, high technical success rate, and satisfactory mid-and long-term outcomes.
4. Research progress on the mutual effects between brown adipose tissue and bone
Mengyue SUN ; Qi YUAN ; Jinglei YANG ; Ridong ZHANG ; Shan TANG ; Sha TAO ; Li MAO
Chinese Journal of Endocrinology and Metabolism 2019;35(11):995-998
Brown adipose tissue (BAT) increases energy consumption by directly dissipating stored energy in the form of heat through the role of uncoupling protein (UCP1). Recent studies have found that brown adipocytes may also regulate metabolism through autocrine, paracrine, and endocrine mechanisms. A growing body of evidences have shown that the BAT has a close relationship with bone metabolism, in which BAT secretes a variety of factors to regulate bone metabolism, while bone also secretes a variety of bioactive substances to control BAT function. In addition, BAT may indirectly participate in bone metabolism through muscle-mediated regulation or SNS activity and improvement of body metabolism, thus forming a BAT-skeletal axis. In this paper, we try to explain the relationship between brown adipose tissue and bone, and to discuss their interactive mechanisms.
5.Effect of tea polyphenois on the growth of human papilIomavirus 16 subgenes-immortalized human cervical epithelial cells
Ziyin MO ; Quan CHEN ; Huaping LI ; Xinyue DAI ; Liqian PENG ; Zhonghao YIN ; Jiusui HUANG ; Bihua LIANG ; Runxiang LI ; Zhenjie LI ; Ridong YANG ; Huilan ZHU
Chinese Journal of Dermatology 2018;51(12):892-896
Objective To explore the effect of tea polyphenols on the growth of human papillomavirus 16 (HPV16) subgenes-immortalized human cervical epithelial cells (H8 cells).Methods Cultured H8 cells were divided into 5 groups to be treated with 0 (control group),6.25,12.5,25 and 50 mg/L tea polyphenols respectively for 24,36,and 48 hours,and then cell counting kit-8 (CCK8)assay was performed to detect cell proliferation.After 24 hours of incubation,flow cytometry was conducted to detect cell apoptosis and cell cycle,and fluorescence microscopy to observe the morphology of apoptotic cells.Results After incubation with tea polyphenols at different concentrations for 24,36 and 48 hours,the proliferation of H8 cells was inhibited,and 12.5 mg/L tea polyphenols could inhibit the relative growth rate of H8 cells in a time-dependent manner.Flow cytometry showed that there was a significant difference in cell apoptosis rate among the 6.25-,12.5-,25-,50-mg/L tea polyphenols groups and the control group (52.62% ± 0.62%,52.22% ± 0.72%,42.52% ± 0.90%,45.96% ± 2.11%,29.96% ± 0.70% respectively,F =272.0,P < 0.05).Moreover,all the tea polyphenol groups showed significantly increased cell apoptosis rate compared with the control group (all P < 0.05).Fluorescence microscopy showed karyopyknosis,nuclear fragmentation and other typical apoptotic morphological changes in H8 cells in tea polyphenols groups.There were significant differences in the percentage of cells in G1,G2 phase and cell proliferation index among the 5 groups (all P < 0.05).Compared with the control group,the 6.25-,12.5-,25-mg/L tea polyphenols groups showed significantly increased percentage of cells in G1 phase (55.96% ± 0.72%,54.12% ± 3.20%,65.30% ± 1.51% respectively,all P < 0.05),but significantly decreased percentage of cells in G2 phase (3.17 ± 1.82%,4.94 ± 1.46%,4.65 ± 4.26% respectively,all P < 0.05) and lower cell proliferation index(0.44 ± 0.01,0.46 ± 0.02,0.36 ± 0.01 respectively,all P < 0.05).Conclusion Tea polyphenols can inhibit the proliferation of H8 cells,induce cell apoptosis,and block cell cycle progression.
6. Effects of cadmium on the expression of estrogen receptor and miRNA in MCF-7 cells
Biyun LI ; Zhipeng LI ; Ridong CAI ; Zhijian CHEN ; Zhihui ZOU ; Rian YU
China Occupational Medicine 2018;45(01):30-34
OBJECTIVE: To study the effects of cadmium on the expression of estrogen receptor( ER) and miRAN-155,miRAN-200 c in human breast cancer MCF-7 cells. METHODS: MCF-7cells in logarithmic growth phase were randomly divided into fulvestrant( ICI182780,ICI) group and non-ICI group. The non-ICI group was treated with cadmium chloride(Cd Cl2) at the final concentrations of 0. 0,2. 5,5. 0 and 10. 0 μmol/L for 24 hours. The ICI group was pretreated at a concentration of 1. 0 μmol/L for 12 hours,and then treated with Cd Cl2 at the final concentrations 0. 0,2. 5,5. 0 and 10. 0μmol/L for 24 hours. The cell proliferation activity was measured by methyl thiazolyl tetrazolium assay. Flow cytometry was used to measured cell apoptosis. Western blot was applied to measure the relative expression of ERα and ERβ protein,and the relative expression of miRNA-155 and miRNA-200 c were detected by real-time fluorescence quantitative polymerase chain reaction. RESULTS: The proliferation rates of MCF-7 cells in 2. 5,5. 0 and 10. 0 μmol/L Cd Cl2 groups were significantly decreased than the 0. 0 μmol/L Cd Cl2 group( P < 0. 05). The proliferation rate in ICI group was lower than that of the non-ICI group( P < 0. 05). When Cd Cl2 concentration was 2. 5,5. 0 and 10. 0 μmol/L,the apoptosis rate of MCF-7 cells in non-ICI group increased compared with those cells without exposure to Cd Cl2( P < 0. 05). The relative expression of ERα,ERβ,miRNA-155 and miRNA-200 c increased( P < 0. 05). The proliferation of MCF-7 cells in ICI group decreased( P < 0. 05),and the relative apoptosis rate increased( P < 0. 05); and the relative expression of ERαand ERβ increased( P < 0. 05),the relative expression of miRNA-155 and miRNA-200 c decreased( P < 0. 05). When treated without Cd Cl2,the apoptosis rate of the ICI group increased compared with non-ICI group(P < 0. 05),the relative expression of ERα and ERβ decreased( P < 0. 05),and the relative expression of miRNA-155 and miRNA-200 c were increased( P < 0. 05). When Cd Cl2 concentration was 2. 5,5. 0 and 10. 0 μmol/L,the apoptosis rate and the relative expression of ERα,ERβ,miRNA-155 and miRNA-200 c decreased compared with the non-ICI group treated with same dose Cd Cl2(P < 0. 05). CONCLUSION: Cadmium can induce cell apoptosis and increase expression of miRNAs through the ER signaling pathway.
7.Genotyping and gene polymorphism of Neisseria gonorrhoeae with azithromycin-resistance and decreased susceptibilities to ceftriaxone
Xiaodong LI ; Jingyao LIANG ; Chao BI ; Ridong YANG ; Ping LI ; Yanhua LIANG ; Xibao ZHANG ; Wenling CAO
International Journal of Laboratory Medicine 2017;38(11):1495-1498,1501
Objective To analyze the characteristics of genotyping and gene polymorphism of Neisseria gonorrhoeae(N.go) with azithromycin(AZM)-resistance(AZM-R) and decreased susceptibility to ceftriaxone(CROD).Methods The minimum inhibitory concentration(MIC) of AZM and CRO were determined.AZM-R isolates were detected for mutations in 23S rRNA,mtrR and penA genes.Genotypes were analyzed by using N.go multi-antigen sequence typing(NG-MAST).Results All total of 485 isolates of N.go were detected.77(15.9%) strains were AZM-R(MIC≥1 mg/L),including 33(6.8%) isolates of AZM low-level resistant(AZM-LLR,MIC=1 mg/L) strains and 44(9.1%) isolates of AZM middle-level resistant(AZM-MLR,MIC≥2 mg/L) strains.There were more CROD(MIC≥0.125 mg/L) strains in AZM-MLR isolates(43.2%),compared with those in AZM-LLR isolates(18.2%,P<0.05).The detected rates of 23S rRNA,mtrR,penA single or combined mutations were without significant differences between AZM-LLR isolates and AZM-MLR isolates(P>0.05).Similar results were found between combined AZM-LLR/CROD isolates and combined AZM-MLR/CROD isolates(P>0.05).No mutation of A2059G and AZM high-level resistant(AZM-HLR,MIC≥256 mg/L) isolate were found.Among 77 AZM-R isolates,67 sequence types(ST) were identified by NG-MAST,of which 30 types were novel.Most ST were represented by a single isolate.Conclusion AZM-R and CROD isolates,presented in this area,might be deserved continuous surveillance to identify the mechanism of concurrent resistance.
8.Heavy metal pollution and health risk in China
Rian YU ; Lingfei HE ; Ridong CAI ; Biyun LI ; Zhipeng LI ; Kedi YANG
Global Health Journal 2017;1(1):47-55
Following rapid social and economic development over the past several decades,pollution by heavy metals has been both serious and widespread in many areas of the world,including China.The situations of heavy metal pollution in China were reviewed,and the health risk and control policy of such pollution were also analyzed and discussed in present paper.
9.Research on Discovery and Application of Regular Syndrome-Treatment Pattern of Classical Chinese Medicinal Formulae for Emotional Diseases Based on Formal Concept Analysis
Sunjing XU ; Saimei LI ; Wenxue HONG ; Zhangzhi ZHU ; Ridong LI ; Yuzhou LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(9):2025-2030
This study was aimed to analyze the regulation of syndrome-treatment pattern of classical Chinese medici-nal formulae for emotional diseases based on formal concept analysis. First, we dealt with the decision formal context of 51 prescriptions about emotional symptom in the Treatise on Febrile and Miscellaneous Diseases and the Es-sentials from the Golden Cabinet based on the principle of optimization. Then, we generated a new partial-order at-tribute diagram in order to present the specific character. Finally, we explained properties of partial-order structure graph from traditional Chinese medicine (TCM) experts' point of view based on knowledge discovery. The results indi-cated the relationship between prescription and syndrome of emotional diseases. It was concluded that method pro-posed in this paper worked well in treatment of description of syndrome differentiation and discovery of new knowl-edge from the known data in the clinical diagnosis.
10.Isolation, expansion and adipogenic differentiation of human keloid-derived precursor cells
Xunxun LIN ; Jie LIU ; Fugui LI ; Ridong WU ; Shi TANG ; Qing TANG
Chinese Journal of Medical Aesthetics and Cosmetology 2013;19(5):368-371
Objective To explore the isolation,amplification methods and adipogenic differentiation under specific culture medium of human keloid-derived precursor cell (KPC) in vitro,in order to study their possibility of being new seed cells of tissue engineering fat.Methods KPCs were isolated from human keloid tissue of 4 different patients in our hospital and were cultured in the modified L-DMEM culture medium.Their cloning efficiency and growth curve were tested.The subcultured cells were tested of the mesenchymal stem cell (MSC)-related gene expression by flow cytometry.In addition,they were cultured in H-DMEM medium (containing 1 μmol/L dexamethasone,0.5 mmol/L 3-isobutyl-1-methyl-xanthine 10 mg/L of bovine insulin,100 mmol/L indomethacin,and 10 % FBS)and were later observed in oil red O staining under phase contrast microscope to determine whether lipid droplets generation was formed,using skin-derived precursors (SKP) as control.Results More than 95 % KPC expressed many antigens of MSC,such as CD29,CD44,CD90 and CD105 while few of them expressed CD34,CD45(1.0 %-2.5 %).And the cells increased in size gradually after inducted the same time,changing from spindle into round or polygonal in shape.The lipid droplets were seen in 72 hours and expressed a positive rate of 78.6 % in Day 19 in oil red O staining while the same rate was 54.6 % in SKP.Conclusions Human keloid-derived precursor cells can express a variety of MSC-related surface markers without expressing hematopoietic stem cell (HSC) related markers.Furthermore,they can be differentiated into fat cells under certain conditions,which may make them as a new source of seed cells for tissue engineering fat.

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