3.Inhibiting PI3K/Akt pathway increases DNA damage of cervical carcinoma HeLa cells by drug radiosensitization.
Shu, XIA ; Shiying, YU ; Qiang, FU ; Fei, LIU ; Wei, ZHENG ; Xiugen, FU ; Yin, ZHAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(3):360-4
This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells. The 50% inhibition concentration (IC(50)) of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay (MTT) in vitro. HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC(20) alone or combined with LY294002 for 24 h, and then radiated by different doses of X-ray. The cell survival ratio was obtained by means of clone formation. One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq), mean lethal dose (D(0)), 2Gy survival fraction (SF(2)) and sensitization enhancement ratio (SER). The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis. The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment (RT) group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group. The apoptosis ratio of each group was measured by flow cytometry. The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells. The Dq, D(0) and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group. The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group, and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group. The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+docetaxel+ RT group was longer than in the cisplatin+RT group or docetaxel+RT group. The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel +RT group was higher than in the cisplatin+RT group or docetaxel+RT group. It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation.
4.Therapeutic effect of HuGanJieXian decoction on rats hepatic fibrosis
Jianchang SHU ; Liang DENG ; Xia Lü ; Yajun HE ; Haiyan ZHU ; Jing FU ; Guorong YE ; Haihua ZHOU
International Journal of Traditional Chinese Medicine 2010;32(3):197-199
Objective To observe therapeutic effects of HuGanJieXian decoction on rats hepatic fibrosis induced by tetrachloride. Methods Rat models of hepatic fibrosis were constructed by intraperitoneal injection of tetrachloride.HuGanJieXian decoction composed of low, middle, and high dose curcumin were given to these rats respectively at the same time. Sho-saiko-to compound treatment group and Fufangbiejiarangan Tablets treatment group were made as positive control groups. After twelve weeks, all rats were executed. Serum samples were kept for measuring serum levels of PC-Ⅲ, LN, and HA. Left livers were extirpated for pathologic examination including H.E and Masson stainings. Grade of hepatic fibrosis were evaluated according to SSS system. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) of supematant centrifugated from hepatic tissue homogenate were detected. Results Serum levels of PC-Ⅲ, LN, and HA were depressed obviously in decoction groups compared with those of fibrotic group (P<0.05) , especially in the low-dose curcumin group.HuGanJieXian Decoction could increase the level of SOD and decrease the level of MDA (P<0.05) , especially in the low-dose curcumin group. Staining of H. E and Masson showed that degrees of hepatic fibrosis in decoction groups were improved obviously compared with that of the fibrotic group. Conclusion HuGanJieXian Decoction can improve rat hepatic fibrosis, the mechanism of this effect may be associated with protecting hepatic cell membrane and anti- peroxidative damage.
5.Role of Sophocarpine in Coxsackievirus B3m Infection in vitro
xing-yu, ZHANG ; shu-xia, CHEN ; jing-xing, LIU ; fu-rong, QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To observe the in vitro anti-Coxsackievirus B3m (CVB3m) effect of sophocarpine(SC) extracted from Sophora flavescens, a traditional Chinese herb. Methods HeLa cells were cultured and the micro-dose cytopathic effect (CPE) assays were applied to detect the toxicity of SC. CPE-inhibitory assays were used to observe the in vitro anti-CVB3m effect of SC. MTT and crystal assays were introduced to examine the anti-CVB3m effect of SC. HeLa cells were infected with CVB3m and added with SC in different concentrations 15 h later.The viability and number of survival of HeLa cells were determined by MTT and crystal violet assays, respectively. Results No toxicity was found on HeLa cells by SC with concentrations 100 ?g/mL, SC could accelerate and aggravate the CPE. SC could protect the CVB3m-infected HeLa cells with concentrations from 1.56 to 25 ?g/mL, and the viability and cell number measured by MTT and crystal violet assay in the SC-handled cells were higher and bigger than those in the virus infected ones. However, the inhibitory effect of virus was exacerbated with higher concentrations (50 and 100 ?g/mL), and the cell number and viability of the SC-handled cells were smaller and lower than those of the infected ones. Conclusion SC with a proper concentration has the in vitro anti-CVB3m effect and may protect HeLa cells from CVB3m infection.
6.Study on Purification and Identification of Streptavidin
Fu-Ying LIU ; Shu-Xia SONG ; Long ZHENG ; Huan-Ling ZHANG ; Hong-Yu YOU ; Jun-Xia WANG ;
Microbiology 1992;0(05):-
The aim of this research is to refine the protocol of purification of SA and identify the character of SA. By utilizing the cold-denaturing method, most of other kinds of protein were screened out and SA was purified from the fermentation broth of L-183 by using the refined affinity chromatography method. The rate of recollection was checked to be 75%~85%. By identification, it is indicated that the molecular weight of self-made SA was 74.5kD, the biotin-combining number 3.2, the activity 11.2u/mg, the pI around 7.4. So, the essential characters of SA are same as described by documents.
7.Investigation of status in coal-burning fluorosis areas in Luoyang city of Henan in 2006
Shu-xia, FU ; Fang-li, YANG ; Jian-shan, KANG ; Juan, MA ; Yu-peng, QIAO ; Qiao-ling, YAO
Chinese Journal of Endemiology 2010;29(2):190-192
Objective To investigate the distribution status of coal-burning fluorosis (endemic fluorosis) areas in Luoyang and to provide scientifc evidence for making strategies in prevention and control. Methods In 2006, a household per village was chosen to carry the general survey so as of disease condition, living habits and housing structure among 941 coal-burning pollution fluorine sickness natural villages in Yanshi, Mengjin, Xin'an, Luanchuan counties and Geely area which were under the jurisdiction of Luoyang. In the general survey, the sampled village having a population of more than 500 person was considered as a major survey village, and water fluoride, 8 - 12 year-old child fluorine spot on tooth and the urinary fluoride were surveyed. Water fluoride and the child urinary fluoride determination used the fluoride ion selective electrode method, and the children's dental fluorosis used Dean method. Results The endemic fluorosis of Luoyang existed 742 in endemic fluomsis villages, compared with history, a decrease of 199 in number. Ninety-six point seven per cent( 142 543/147 419) of the households were consuming smoke-free coal. Households using intact kitchens accounted for 93.6%( 137 919/147 419). Of which 63.0%(86 889/137 919) of kitchens were mixed up with bedrooms. Total 125 060 people were using coal- fired furnace for heating, of which 87.8%(109 802/125 060) had smoke-free facilities, 12.2%(15 258/125 060) had none. Among 52 endemic villages with population of more than 500 people surveyed, a total of 183 water samples were collected, 2 had water fluoride exceeding 1.0 rag/L, the highest water fluoride being 1.04 rag/L, averaging 0.39 mg/L Sixteen villages had a prevalence rate of dental fluorosis for children less than 30.00%, accounting for 30.8% (16/52), 36 endemic villages the prevalence of dental fluorosis detection rate of more than 30.00%, accounting for 69.2%(36/52). Twenty-thrce villages had a dental fluorosis index greater than 0.6, severe dental fluorosis was not found. Real-time measurement of 1408 urine samples of children aged 8 - 12 showed that urine fluoride highest value 6.88 nag/L, the minimum value of 0.10 mg/L, geometric mean 1.10 mg/L. The prevalence rate of dental fluorosis for children was 36.06%. Conclusions In Luoyang city, numbers of coal-burning endemic fluorosis villages are less than before, children's dental fluorosis has significantly declined, however some people still use kitchens connecting with bedrooms and lack smoke-free facilities, they need to be educated to change lifestyle and improve furnace to reduce soot fluoride pollution.
8.Inhibiting PI3K/Akt Pathway Increases DNA Damage of Cervical Carcinoma HeLa Cells by Drug Radiosensitization
XIA SHU ; YU SHIYING ; FU QIANG ; LIU FEI ; ZHENG WEI ; FU XIUGEN ; ZHAO YIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(3):360-364
This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells.The 50% inhibition concentration(IC50)of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay(MTT)in vitro.HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC20 alone or combined with LY294002 for 24 h,and then radiated by different doses of X-ray.The cell survival ratio was obtained by means of clone formation.One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq),mean lethal dose(D0),2Gy survival fraction(SF2)and sensitization enhancement ratio(SER).The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis.The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment(RT)group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group.The apoptosis ratio of each group was measured by flow cytometry.The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells.The Dq,Do and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group.The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group,and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group.The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+do-cetaxel+RT group was longer than in the cisplatin+RT group or docetaxel+RT group.The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel+RT group was higher than in the cisplatin+RT group or docetaxel+RT group.It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation.
9.Isolation and identification of steroidal saponins in total saponin from Dioscorea nipponica Makino.
Shu-hu DU ; Wen-ying LIU ; Tie-jun FU ; Bo-gang XIA ; Chong-dao XIA
Acta Pharmaceutica Sinica 2002;37(4):267-270
AIMTo investigate the water-soluble steroidal saponins in total saponin from Dioscorea nipponica Makino and look for new active compounds.
METHODSThe compounds were isolated with silica gel, PTLC and HPLC, and their structures were elucidated by acid hydrolysis, physical and chemical data and spectral analysis (IR, NMR, MS, HMQC, HMBC) as well as chemical correlations.
RESULTSThe two steroidal saponins (water-insoluble saponin and water-soluble saponin) were isolated from the total saponin of Dioscorea nipponica Makino. The structures were elucidated as diosgenin 3-O-[alpha-L-rhamnopy ranosyl (1-->2)-[beta-D-glucopyranosyl(1-->3)]]-beta-D-glucopyranoside (I), diosgenin 3-O-[alpha-L-rhamnopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl (1-->4)]-beta-D-glucopyranoside (II).
CONCLUSIONCompound II is a new steroidal saponin and firstly isolated from Dioscorea nipponica Makino. It was named as dioscin Dc.
Dioscorea ; chemistry ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification
10.Tumor formation of prostate cancer cells influenced by stromal cells from the transitional or peripheral zones of the normal prostate.
Fu-Jun ZHAO ; Bang-Min HAN ; Sheng-Qiang YU ; Shu-Jie XIA
Asian Journal of Andrology 2009;11(2):176-182
This study was designed to investigate the different involvements of prostatic stromal cells from the normal transitional zone (TZ) or peripheral zone (PZ) in the carcinogenesis of prostate cancer (PCa) epithelial cells (PC-3) in vitro and in vivo co-culture models. Ultra-structures and gene expression profiles of primary cultures of human prostatic stromal cells from the normal TZ or PZ were analyzed by electron microscopy and microarray analysis. In vitro and in vivo co-culture models composed of normal TZ or PZ stromal cells and human PCa PC-3 cells were established. We assessed tumor growth and weight in the in vivo nude mice model. There are morphological and ultra-structural differences in stromal cells from TZ and PZ of the normal prostate. In all, 514 differentially expressed genes were selected by microarray analysis; 483 genes were more highly expressed in stromal cells from TZ and 31 were more highly expressed in those from PZ. Co-culture with PZ stromal cells and transforming growth factor-beta1 (TGF-beta1) increased the tumor growth of PC-3 cells in vitro and in vivo, as well as Bcl-2 expression. On the other hand, stromal cells of TZ suppressed PC-3 cell tumor growth in the mouse model. We conclude that ultra-structures and gene expression differ between the stromal cells from TZ or PZ of the normal prostate, and stroma-epithelium interactions from TZ or PZ might be responsible for the distinct zonal localization of prostate tumor formation.
Adenocarcinoma
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drug therapy
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genetics
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pathology
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Adult
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Animals
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Cell Line, Tumor
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Coculture Techniques
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Gene Expression
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drug effects
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Gene Expression Profiling
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Humans
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Male
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Mice
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Mice, Nude
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Oligonucleotide Array Sequence Analysis
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Prostate
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drug effects
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metabolism
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pathology
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Prostatic Neoplasms
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drug therapy
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genetics
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pathology
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Proto-Oncogene Proteins c-bcl-2
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genetics
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metabolism
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RNA, Messenger
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metabolism
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Stromal Cells
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metabolism
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pathology
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Transforming Growth Factor beta
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pharmacology
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Young Adult