1.A preliminary study of serum-deprivation-induced release of S100A13 and flbroblast growth factor 1 from thyroid cancer TT cell
Jingjin YANG ; Fang WEN ; Renxian CAO ; Jing ZHONG ; Gebo WEN
Chinese Journal of Endocrinology and Metabolism 2011;27(11):941-944
Objective To investigate the effect of serum-deprivation on the changes of [ Ca2+ ] i and the protein release of S100A13 and fibroblast growth factor 1 ( FGF-1 ) in thyroid cancer TT cell,and to reveal the role of Ca2+in the protein release of S100Al3 and FGF-1.Methods The protein expressions of FGF-1 and S100A13 in TT cells under serum-deprivation were detected by Western blot.The released FGF-1 protein from TT cells in the supernatant fluid was detected by ELISA.Realtime dynamic examinations on the change of 1 h [ Ca2+ ] i in TT cells under serum-deprivation were detected by confocal laser scanning microscopy.Then,the effect of EGTA( 2.5 mmol/ L),BAPTA-AM (2.5 μmol/L)on distributions of the fluorescence of S100AI 3 and FGF-1 in TT cells under serumdeprivation for6 h were detected by indirect immunofluorescence.Results The expressions of FGF-1 and S100A13 in TT cells after serum-deprivation for4 h and 6 h were reduced( P<0.05 or P<0.01 ),but the released FGF-1 protein from TT cells in the supernatant fluid was elevated ( P<0.05 or P<0.01).Confocal laser scanning of Ca2+ imaging indicated that [ Ca2+ ] i of serum-deprivation TT cells maintained the relative stabilization within 23 win,but the rapid rise of [ Ca2+ ] i achieved peak value 1.6 μmol/L after 30 min,and remained stable for about 17 win,and thereafter 40 win slowly dropped to a low level From 40 win to 60 win the [ Ca2+ ] i was about 0.3-0.6 μ mol/L.The average [ Ca2+ ] i was higher than that in normal group,EGTA group,and BAPTA-AM group within 1 h.The protein expressions of S100A13 and FGF-1 did not drop obviously in EGTA group and BAPTA-AM group.Conlusion The release of S100A13 and FGF-1 from TT cell under serum-deprivation is possibly related with the change of [ Ca2+ ]i.Both Ca2+-chelating agents EGTA and BA PTA-AM are able to inhibit the rise of [ Ca2+ ] i and release of S100A 13 and FGF-1 from TT cells under serum-deprivation.
3.Progress of Epitope Vaccine
Zhong FANG ; Wen-Xin LUO ; Ning-Shao XIA ;
China Biotechnology 2006;0(11):-
Epitope vaccine is one of the emerging vaccine techniques developing in past decade years.Particularly the advantages of this vaccine on preventing and therapy illness,as cancer and virus,are espacially outstanding.The most importent elements about the vaccine,namely T/B-epitopes obtainment and identification,vehicles for epitope and vaccine construction,are reviewed.In addition,applications of the vaccine technique in some refractory diseases,such as cancer,virus and pathogen infection,are depicted.
4.Effects of small hairpin RNA-mediated S100A13 gene inhibition on the release of fibroblast growth factor-1 in human thyroid cancer cells
Lina TIAN ; Renxian CAO ; Xing LIU ; Fang WEN ; Jing ZHONG ; Bin YAN ; Gebo WEN
Chinese Journal of Endocrinology and Metabolism 2010;26(10):847-849
Objective To investigate whether the release of fibroblast growth factor-1 ( FGF-1 ) was changed after inhibition of S100A13 gene (small hairpin RNA, shRNA)and serum-deprivation in human thyroid cancer cells (TT cells ). Methods The S100A13-shRNA pENTRTM/U6 entry vector was transfected into TT cells. The expression of S100A13 mRNA and protein was detected by immunoflurescence, real-time RT-PCR, and Western blot. Then TT cells were treated with S100A13 gene inhibition and serum-deprivation. The changes in release of FGF-1 were detected by indirect immunoflurescence, RT-PCR, and ELISA. Results S100A13 shRNA transfected TT cells (S100A13 RNAi cells)had a reduction of S100A13 gene and protein expression by 80%.Indirect immunofluorescence indicated FGF-1 was mostly localized in the cytoplasm and nucleus of TT cells in primary culture. When serum-deprivation stress was given to TT cells, FGF-1 in cytoplasm almost disappeared in the cells at 6 h. RT-PCR indicated that when serum-deprivation stress was given to TT cells the mRNA of FGF-1 was reduced. ELISA showed that with inhibition of S100A13, the release of FGF-1 was reduced (P<0.05).Conclusion S100A13-shRNA pENTRTM/U6 entry vector transfected TT cells may inhibit the expression of S100A13 and reduce the release of FGF-1.
5.Determination of 27 elements in Maca nationality's medicine by microwave digestion ICP-MS.
Gui-fang YU ; Hai-jie ZHONG ; Jun-hua HU ; Jing WANG ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(23):4545-4551
An analysis method has been established to test 27 elements (Li, Be, B, Mg, Al, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Sr, Mo, Cd, Sn, Sb, Ba, La, Hg, Pb, Bi) in Maca nationality's medicine with microwave digestion-ICP-MS. Sample solutions were analyzed by ICP-MS after microwave digestion, and the contents of elements were calculated according to their calibration curves, and internal standard method was adopted to reduce matrix effect and other interference effects. The experimental results showed that the linear relations of all the elements were very good; the correlation coefficient (r) was 0.9994-1.0000 (Hg was 0.9982) ; the limits of detection were 0.003-2.662 microg x L(-1); the relative standard deviations for all elements of reproducibility were lower than 5% (except the individual elements); the recovery rate were 78.5%-123.7% with RSD lower than 5% ( except the individual elements). The analytical results of standard material showed acceptable agreement with the certified values. This method was applicable to determinate the contents of multi-elements in Maca which had a high sensitivity, good specificity and good repeatability, and provide basis for the quality control of Maca.
Lepidium
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chemistry
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methods
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Reproducibility of Results
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Trace Elements
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chemistry
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isolation & purification
6.Comparison of three neoadjuvant chemotherapy in treating cervical carcinoma
Qian ZHONG ; Wanmin LIU ; Xiaoqin LIAN ; Haiqing WANG ; Weijian ZHANG ; Fang LIU ; Baoning WEN
Chinese Journal of Postgraduates of Medicine 2010;33(30):23-25
Objective To explore the application value of neoadjuvant chemotherapy (NACT) in treating cervical carcinoma. Methods Seventy-one cervical carcinoma patients who were divided into three groups:PF group (cisplatin and fluorouracil,23 cases) or CBP group (carboplatin,bleomycetin and cyclophosphamide, 24 cases) or TP group (paclitaxel and cisplatin, 24 cases); operation was made 14 - 21 days afterwards. The therapeutic effect, chemotherapy side-effect and the effect on operation and pathology for these three groups were analyzed and compared. Results The effective rate was 93.8%(30/32),88.6% (31/35),82.4%(28/34) in TP group,CBP group,PF group,respectively. However, the therapeutic effect had no statistics significance with age, preoperative tumor grade, pathologic type for all the three groups, the therapeutic effect had statistical significance with clinical stage. The rate which pathological examination showed no residual cancer was biggest in CBP group [CBP group was 28.6%(6/21), TP group was 4.3% (1/23), PF group was 11.8%(2/17), P < 0.05 ]. Conclusions Three NACT projects are safe and effective treatment for cervical cancer. However,each project has advantages. The NACT projects can be elected for different patient according to his illness and economy.
7.Study on the rhythm of urine iodine level of children aged 8-10 in Chongqing city
Ting, ZHANG ; Ge, LI ; Bang-zhong, XIAO ; Wen-fang, LIAO ; Xin-shu, LI ; Gui-wang, DOU
Chinese Journal of Endemiology 2010;29(3):313-315
Objective To undemtand the rhythm of urinary iodine level of children aged 8-10 in Chongqing city.Methods In April 2008,using the stratified random sampling method,we sampled 60 children aged 8-10 in a lodging primary school in Chongqing(20 per age group,half male and half female),the urine samples were collected in the morning and at 10:00,12:30,16:00,iodine in urine was detected by method of Ce and arsenic catalytic speetrophotometry(WS/T 107-2006).The difference of the urinary iodine level was compared by age,sex and time of day.Results The median urinary iodine of 60 children was 265.07μg/L on the overall.Irrespective of the stratification factors,excluding morning urinary iodine(366.75μg/L)and urinary iodine at 10:00(338.30 μg/L),the urinary iodine between 12:30(235.15μg/L)and 16:00(251.50μg/L)was not significant(all P>0.05),statistically significant differences(all P<0.05)were found between any two.The urinary iodine of 8-year-old group at different times of the day was significantly different(all P<0.05),except between morning urinary iodine (298.90 μg/L)and at 10:00,16:00(279.00,286.59 μg/L),between urinary iodine at 10:00 and 16:00(all P>0.05).The 9-year-old group's urinary iodine were not significantly different between morning urine(366.15μg/L)and 10:00(368.10 μg/L),and between 12:30(244.00 μg/L)and 16:00(186.30 μg/L,all P>0.05),significant differences were faund at other times of the day(all P<0.05).The 10-year-old group of urinary iodine changed very little before 12:30 (382.85,449.60,337.00 μg/L, all P > 0.05 ), followed by rapid decline to 16: 00 (269.35 μg/L), and compared with the morning urine and 10:00, there was significant difference(all P < 0.05).Regardless boys or girls, the urinary iodine at different times qf the day was significantly different (all P < 0.05),except between morning urinary iodine(337.32,309.28 μg/L) and at 10:00(316.15,288.27 μg/L), between urinary iodine at 12:30(251.18,211.45 μg/L) and 16:00(235.02,211.45 μg/L, all P > 0.05). Conclusions The change of urinary iodine level in children aged 8 - 10 was not obvious before noon, changes can be seen in the afternoon.Urinary iodine level before 10:00 is indicative.
8.A comparison of four methods for extraction of human fecal DNA by using real time PCR
Zhong-Wen WU ; Ying HAN ; Hai-Feng LU ; Lan-Juan LI ; Ji-Fang SHENG ; Jian ZUO ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P
9.EFFECT OF r-IRRADIATION ON THE METABOLISM OF ASCORBIC ACID
Yun-Zhong FANG ; Pei-Gang WANG ; Ye-Fu LAI ; Rui-Wen LI ;
Acta Nutrimenta Sinica 1956;0(01):-
Guinea pigs were given orally with a daily dose of 0.5 mg ascorbic acid per 100 g body weight 20 days before and 4 days after a whole-body r-irradiation and then injected subcutaneously with 10 mg ascorbic acid at 24 hours intervals. During the injection period, the daily urinary output of total ascorbic acid in the animals irradiated with 500 or 1000 r was significantly higher than that in controls, but the difference of 24 hours excretions of total ascorbic acid between the group irradiated with 150 or 250 r and the control group was not statistically significant.As shown in the urinary output of dehydroascorbic acid and diketo- gulonic acid estimated by our modified Roe's method, r-irradiation has no effect on its excretion in guinea pigs and rats.The levels of ascorbic acid in plasma, spleen and adrenals were significantly lower in the irradiated group than in the control group, but might increase with the increase of the injected dose of ascorbic acid.
10."Treating Diabetes Mellitus from the Viewpoint of Spleen: Based on the Theory of ""Homogeny of Spleen and Pancreas"""
Jun CHAO ; Ya LIU ; Wen ZHONG ; Chuanming FANG ; Chunguang XIE ; Hong GAO
Journal of Traditional Chinese Medicine 2017;58(17):1458-1461
The academic origin of homogeny of spleen and pancreas is explained from the aspect of Chinese medicine.The authors think spleen faihng to spread essence is the basic pathogenesis to diabetes mellitus.Spleen function of spreading essence is impaired.Thus essence of water and grain could not be spread in the whole body but amass sugar-turbidity,which manifests high blood sugar.Differentiating diabetes mellitus from the viewpoint of spleen,invigorating spleen and benefiting Qi could help spleen to ascend clear.Invigorating spleen-yin and clearing endogenous heat are used.The liver and kidney should be considered.The methods of dissipating phlegm and activating blood circulation could be combined.The treating idea of treating spleen is treating pancreas should be used in preventing and treating diabetes mellitus.