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.Expression of uncoupling protein-2 mRNA in diet-induced obesity-resistant rats.
Rong LIU ; Chang-hao SUN ; Rong-xia FU ; Yue-shan YAN ; Shu-ran WANG
Chinese Journal of Preventive Medicine 2003;37(6):446-449
OBJECTIVETo explore the expression of uncoupling protein-2 mRNA in brown adipose tissue, white adipose tissue and skeletal muscle of diet-induced obesity-resistant (DIO-R) rats.
METHODSFifty male Sprague-Dawley (SD) rats were randomly divided into a control group and a high-fat group and fed with basic diet and high-fat diet respectively for 13 weeks. DIO-R and DIO rats were selected according to their body weight. The change of body weight and the intake of total calorie were observed. Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of UCP2 mRNA in rat.
RESULTSBody weight and total calorie intake in DIO-R rats (425.1 +/- 27.1) g, (31,693 +/- 946) kJ were significantly lower than those in DIO rats (489.7 +/- 20.5) g, (34,363 +/- 1465) kJ. The peak area of UCP2 mRNA in white adipose tissue in DIO-R rats was 352 +/- 30 and in DIO rats was 101 +/- 12. The peak areas of UCP2 mRNA in skeletal muscle in DIO-R and DIO rats were 130 +/- 15 and 170 +/- 12, respectively. The peak areas of UCP2 mRNA in brown adipose tissue of DIO and DIO-R rats were 124 +/- 14 and 147 +/- 19, respectively.
CONCLUSIONThe expression of UCP2 mRNA in white adipose tissue of DIO-R rats increased significantly. These results suggest that obesity-resistance was associated with a tissue-specific increase in UCP2 expression.
Adipose Tissue ; drug effects ; metabolism ; Animals ; Body Weight ; drug effects ; Dietary Fats ; administration & dosage ; Gene Expression ; genetics ; Ion Channels ; Male ; Membrane Transport Proteins ; genetics ; Mitochondrial Proteins ; genetics ; Muscle, Skeletal ; drug effects ; metabolism ; Obesity ; etiology ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Uncoupling Protein 2