3.Silencing of PSMA by RNAi influences biological behavior of prostate cancer cell line LNCaP
Kaiyuan CAO ; Tian ZHANG ; Lin XU ; Guangqing YUAN ; Xiaodong TIAN ; Xiaorong HUANG ; Shaopeng QIU
Chinese Journal of Pathophysiology 2000;0(10):-
AIM:To study the blocking effect of shRNA on the expression of PSMA gene in LNCaP cell line by using shRNA eukaryotic expression vector.METHODS:Three pairs of DNA templates coding shRNA,synthesized against PSMA and cloned into the vector pSilencer 2.1-U6-neo,which was named pSilencer 2.1-U6-neo-shRNA,were identified by restriction endonuclease digestion analysis and DNA sequencing.LNCaP cells were then transfected with these three pSilencer 2.1-U6-neo-shRNAs and the negative control pSilencer 2.1-U6-neo-NC.After G418 selection,the cells were selected and the interfering effect was detected by RT-PCR and Western blotting.The biological behaviours of the transfected LNCaP cells were also tested.RESULTS:Restriction endonuclease digestion analysis and DNA sequencing results all showed that the 3 target segments were cloned into pSilencer 2.1-U6-neo vector respectively.After transfected into LNCaP cells,the inhibitory ratio of PSMA mRNA was 33.15%,9.26% and 41.97% respectively,and that of PSMA protein was 26.26%,6.47%,40.69% respectively.The p-shRNA3 was chosen to test the cell growth and its invasive power in vitro.The results showed that after interfering,the invasiveness of LNCaP cells were enhanced.CONCLUSION:The vector-based shRNA on PSMA gene effectively knocks down the PSMA gene expression.The successful construction of PSMA shRNA makes it possible for further study of the interaction between PSMA and prostate cancer.
4.Analysis on morphological characteristics, photosynthetic characteristics and chemical constituents of Inula lineariifolia from different populations.
Feng-chen CHEN ; Chang-lin WANG ; Qiao-sheng GUO ; Xin TIAN ; Yuan-yuan QIU
China Journal of Chinese Materia Medica 2015;40(22):4389-4394
Twelve populations of Inula lineariifolia were used as materials to measure morphological characteristics, photosynthetic parameters and chemical constituents. It aims to provide a theoretical basis for germplasm resources evaluation. The results showed that I. lineariifolia had relatively rich morphological diversity, there were significant differences of morphological characteristics, photosynthetic parameters and chemical constituents among populations. There was positive correlation on morphological characteristics and P(n). Twelve populations were divided into three-type. The three populations of Xuyi, Mingguang and Fengyang were of narrower-longer leaf, bigger biomass,better photosynthetic and higher chemical constituents. Then they were classified for a similar group. It proved that the three populations were more suitable for cultivation and promotion.
Biomass
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China
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Flowers
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chemistry
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growth & development
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metabolism
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Inula
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chemistry
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classification
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growth & development
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metabolism
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Photosynthesis
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Plant Leaves
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chemistry
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growth & development
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metabolism
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Plant Stems
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chemistry
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growth & development
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metabolism
5.The role of histone H3 acetylation on cleft palate in mice induced by 2, 3, 7, 8-tetrachlorodibenzopdioxin .
Liu CUIPING ; Yuan XINGANG ; Fu YUEXIAN ; Qiu LIN ; Tian XIAOFEI ; Liu YAN ; Wei GUANGHUI
Chinese Journal of Plastic Surgery 2014;30(5):369-372
OBJECTIVETo explore the role of histone H3 acetylation in cleft palate induced by 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD) in C57BL/6J mice, and its mechanism.
METHODSOn gestation day 10 (GD10), 36 pregnant mice were randomly divided into two groups as the treated group(n = 18) and the control group( n = 18). The mice in the treated group received intragastric administration with TCDD 28 μg/kg, while the mice in the control group received equivalent corn oil. The pregnant mice were sacrificed on GD13. 5, GD14. 5 and GD15. 5, collecting fetal palates to determine the activities of histone acetyltransferases (HATs) by Colorimetric and the expression level of acetylated histone H3 (Acetylated histone H3, Ac-H3) by Western-blot.
RESULTSThe activity of HATs was 0.409 7 ± 0.0147, 0.522 3 ± 0.017 1 and 0.643 5 ± 0.013 9 in control group on GD13.5, GD14.5 and GD15.5; 0.865 0 ± 0.0129, 0.719 1 ± 0.017 8 and 0.551 2 ± 0.016 8 in TCDD group. The activity of HATs in TCDD group was higher than that in control group on GD13. 5, GD14. 5, showing significantly difference between the two groups (t = - 56. 932, t = - 19. 516, P < 0.01); however, the activity of HATs in TCDD group was significantly lower than that in control group on GD15. 5 (t = 10. 382, P < 0.01). The expression level of Ac-H3 was 0.745 0 ± 0.113 5, 1.055 9 ± 0.249 4 and 1.795 5 ± 0.081 9 in control group on GD13. 5, GD14. 5 and GD15. 5; while 1.4490 ± 0. 1460, 1. 641 8 ± 0.099 7 and 1. 512 1 ± 0. 150 2 in TCDD group. The expression of Ac-H3 in TCDD group was higher than that in control group on GD13. 5, GD14. 5, showing significantly difference( t = -6. 593, -3. 779, P <0. 01, P <0.05) ; However, the expression of Ac-H3 in TCDD group was statistically lower than that in control group (t = 2. 870, P <0. 05).
CONCLUSIONThe acetylation of histone H3 was involved in the cleft palate of C57BL/6J mice induced by TCDD, which may be one of the mechanisms in TCDD-induced cleft palate.
Acetylation ; drug effects ; Acetyltransferases ; metabolism ; Animals ; Cleft Palate ; chemically induced ; metabolism ; Dioxins ; Female ; Fetus ; Histones ; metabolism ; Humans ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; Pregnancy ; Random Allocation ; Teratogens
6.Reconstruction finger web with dorsal two wing-shaped flap for the treatment of congenital syndactyly.
Xiaofei TIAN ; Lin QIU ; Yuexian FU ; Yan LIU ; Xinguang YUAN ; Jun XIAO ; Tianwu LI
Chinese Journal of Plastic Surgery 2014;30(2):96-98
OBJECTIVETo explore the clinical effect of using dorsal two wing-shaped flap to reconstruct finger web for treatment of congenital syndactyly.
METHODSThis technique has been used in 19 children with congenital syndactyly. At the dorsum, a flap with V-shaped tip and two wing-shaped pedicle were designed and was just sewed up with an anchor-shaped incision at the palm. The web was primarily reconstructed without skin graft at base of fingers. Distal end of fingers were separated by using serrated flap and were closed after removal of fatty tissue. At some cases with tight skin connection. The defect area at lateral and distal end of fingers was closed by small pieces of skin graft.
RESULTSAll the webs were reconstructed primarily without skin graft at the base of fingers. 7 cases with tight skin connection had small pieces of skin graft at lateral and distal end of fingers. Primary healing was achieved in all cases. After 1 to 6 months of follow-up, both the appearance and function were satisfactory without conspicuous scar. The reconstructed finger webs were in normal depth and width.
CONCLUSIONSPrimary web space can be achieved by dorsal two wing-shaped flap without skin graft at base of fingers. It is one of the best choices for treatment of congenital syndactyly.
Adipose Tissue ; surgery ; Child ; Cicatrix ; Dermatologic Surgical Procedures ; methods ; Fingers ; surgery ; Humans ; Skin Transplantation ; Surgical Flaps ; transplantation ; Syndactyly ; surgery ; Wound Healing
7.Effects of p38MAPK signaling pathway on cyclic tensile stress-induced fibroblast apoptosis
Jing QIU ; Guangyun ZHANG ; Zhen TIAN ; Yue ZHANG ; Jiangbo YU ; Xiao YUAN
Chinese Journal of Tissue Engineering Research 2011;15(20):3789-3792
BACKGROUND: When the teeth affected abnormal biting force, tooth absorption and periodontium would be greatly damaged. OBJECTIVE: To study whether periodontal membrane fibroblast affected apoptosis following cyclic tensile stress stimulation and whether p38MAPK signaling pathway participated in apoptosis. METHODS: Fibroblasts at passages from 4 to 7 were randomly assigned to control, loading and SB203580 groups after synchronization. In the loading and SB203580 groups, 12% strain was applied at a loading frequency of 6 cycles per minute, i.e. 5 seconds for tension and 5 seconds for relaxation. In the SB203580 group, cells were treated with 20 mmol/L p38MAPK inhibitor SB203580 at 1 hour before loading. At 6, 12 and 24 hours after loading, cells from each group were harvested, and cell apoptosis was detected using a flow cytometry. Expression of bax mRNA was determined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Compared with the control group, apoptotic rate of fibroblasts and bax mRNA expression were increased after loading (P < 0.05), and enhanced over time, and peaked at 12 hour following loading, and then decreased gradually. Compared with the loading group, cell apoptosis was reduced at corresponding time points in the SB203580 group (P < 0.05), and bax mRNA expression was diminished. These results indicated that cells affected apoptosis after mechanics stimulation, and mitogen activated protein kinase p38MAPK signaling pathway participates in the process of apoptosis.
8.Regularity and predictors of weight loss during concurrent chemoradiotherapy for nasopharyngeal carcino- ma patients
Ronghui ZHENG ; Yingying LIANG ; Yunhong TIAN ; Xiubi GUAN ; Huizhi QIU ; Anan XU ; Weijun ZHANG ; Yawei YUAN
The Journal of Practical Medicine 2017;33(16):2682-2685
Objective To investigate the regularity and predictors of weight loss during concurrent chemo-radiotherapy for nasopharyngeal carcinoma patients. Methods A retrospective study was carried out on 82 patients with nasopharyngeal carcinoma ,with average age of 45 years ,who received concurrent chemoradiotherapy at the Affiliated Cancer Hospital of Guangzhou Medical University. Paired t-test was used to analyze the weight loss pattern during chemoradiotherapy. The Spearman′s rank correlation analysis was used to investigate the correlation between high weight loss(HWL)and the clinical factors. Results The paired t-test showed that the weight of the patients decreased continuously every week during chemoradiotherapy(P < 0.05),and the value dropped most significantly during the 4th week. Among these 82 patients,32 were determined as HWL(weight loss > 10%). Spearman′s rank correlation analysis showed that HWL was associated with older age,female,higher T and TNM stage,3D-CRT technology,the higher dose of radiotherapy in nasopharynx and in neck lymph node,and decreases of leukopenia and hemoglobin over degree Ⅱ(P<0.05). Conlousions Weight may decrease continuously every week during chemoradiotherapy for nasopharyngeal carcinoma patients. Age,gender,T stage,TNM stage,radio-therapy dose,and leukopenia and hemoglobin decreases may be predictors for weight loss degree during concurrent chemoradiotherapy for nasopharyngeal carcinoma patients.
9.Analysis of gene structure and diversity of new alternative spliced variants of prostate specific membrane antigen
Kaiyuan CAO ; Na XIAO ; Lin XU ; Guangqing YUAN ; Shuqin DAI ; Xiaorong HUANG ; Xiaodong TIAN ; Shaopeng QIU
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.
10.Correlation between Pulse Pressure and Cardiovascular Structure and Function in Aged Patients with Essential Hypertension
qiu-rong, MENG ; ning-yuan, FANG ; tian, GAO ; lin-qiao, LI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(09):-
Objective To study the correlation between pulse pressure(PP) and cardiovascular structure and function in aged patients with essential hypertension. Methods Forty-eight aged patients with essential hypertension were enrolled into the study and divided into the following two groups by the mean value of PP: PP