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.
2.The influence and screening value of percentage of body fat and waist height ratio for hyperlipidemia in physical examination people
Yongchun CHEN ; Xingguo CAO ; Jing WEN ; Xueting LI ; Lin SUN
Tianjin Medical Journal 2015;(8):916-919
Objective To evaluate the relationship and screening value of percentage of body fat (BF%) and waist height ratio (WHtR) for hyperlipidemia in physical examination people. Methods A total of 2 668 objects taking physical examination in Henan Province People′s Hospital from September to December 2014 were included in this study. Values of height, body mass index (BMI), waist circumference, body composition and blood lipid level were detected. The receiver oper?ating characteristic curve (ROC) was used to analyze the screening rate of WHtR and BF%on hyperlipidemia with sensitivi?ty, specific and area under the curve (AUC). After stratified by age, waist circumference and BMI, multivariable logistic re?gression analysis was used to investigate the association between hyperlipidemia risk, BF% and WHtR. Results The screening accuracy rate on hyperlipidemia was higher for BF%, AUC was 0.79 in both female and male people. Among wom?en with BMI<18.5 kg/m2 and 18.5~<24 kg/m2, the risk of hyperlipidemia was higher in superfatted group than that of normal group. There was no correlation between WHtR and hyperlipidemia. Among men older than 40 y or with abnormal waist cir?cumference (≥85 cm), the risk of hyperlipidemia was higher in superfatted group than that of normal group, but not associat?ed with WHtR. Conclusion The BF%is a better screening indicator for hyperlipidemia compared with that of WHtR and BMI. Women with BMI<18.5 kg/m2 and 18.5~<24 kg/m2 and men older than 40 y or with waist circumference over 85 cm are suggested to do body composition tests to improve screening accuracy for hyperlipidemia.
3.The effect of warm ischemia duration on renal function early after laparoscopic partial nephrectomy
Jing CAO ; Wenjun CHEN ; Jiaming WEN ; Jingang ZHAO ; Chuanjun DU
Chinese Journal of Urology 2015;(6):414-418
Objective To assess the effect of variable durations of warm ischemia on renal function early after laparoscopic partial nephrectomy ( LPN ) and make the definite safety duration of renal warm ischemia.Methods The clinical data of 76 patients treated with LPN from October 2012 to June 2014 were retrospectively analyzed.The patients were divided into 3 groups based on warm ischemic time,namely group A (28 cases) with warm ischemia time less than 20 min,group B (34 cases) with warm ischemia time more than 20 min and less than 30 min, group C ( 14 cases ) with warm ischemia time more than 30 min.LPN was performed with renal artery clamping alone in all the patients.Preoperative and postoperative renal scintigraphic scan was performed to access glomerular filtration rate ( GFR) in all patients.The GFR values were compared among before, 1 week, 1 month and 3 months after operation.The factors predicting the early renal injury were assessed by multivariate regression analysis.Results The renal GFR of the kidney underwent LPN decreased 19.43(17.70,22.06) ml/min at 1 week,17.04(13.94,20.70) ml/min at 1 month,13.82(10.72,18.73) ml/min at 3 months after the surgery in group C,respectively.In group A, the renal GFR of the operated-side decreased 12.07(10.91,13.42) ml/min,10.04(9.16,11.75) ml/min, 8.44(7.07,9.72) ml/min,respectively.In group B, the renal GFR of the operated-side decreased 13.64 (12.48,16.72) ml/min,10.29(9.17,14.27)ml/min,9.63(7.85,12.59) ml/min,respectively.The GFR decreased greater in group C than that in group A and B(P<0.05).The total renal GFR decreased (10.70 ± 4.93)ml/min at three months in group C,compared with (5.64 ±4.12)ml/min in group A and (6.37 ± 4.32)ml/min in group B,respectively.The decreased value in group C was greater than that in group A and B(P<0.05).However,the differences of the total renal GFR among the 3 groups were not significant at 1 week and 1 month(P>0.05).The multivariate regression analysis revealed that warm ischemia duration was the independent risk factor of the early renal injury.Conclusions Warm ischemia duration is the major factor regarding the early renal recovery after LPN.Warm ischemia time more than 30 min may not only greatly affect the renal function but also the renal function recovery rate.
4.Effects of mild hypothermia on cardiac function in early stage of post-resuscitation in rabbits
Wen CAO ; Peijie LI ; Liping ZHANG ; Jing QING ; Yangling LI
Chinese Journal of Emergency Medicine 2012;21(6):622-625
ObjectiveTo observe the effects of mild hypothermia on post-resuscitation myocardial dysfunction in rabbits in order to elucidate the underlying mechanism of hypothermia.Methods After setting up rabbit model of cardiopulmonary resuscitation,20 rabbits were randomly ( random number)divided into two groups,namely normothermic resuscitation group (group A,n =10 ) and post-ROSC hypothermia group ( group B,n =10).In the group A,animals wore treated with standard CPR after cardiac arrest.In post-ROSC hypothermia group,the body temperature of animals was cooled to 32 ~ 34°C after successful ROSC.The left ventricular end-diastolic pressure (LVEDP),left ventricular pressure rise and fall rates ( ± dp/dtmax,serum concentrations of heart-type fatty acid-binding protein (H-FABP) and 8-isoprostaglandin F2a (8-iso-PGF2a) and Cyclooxygenase-2 (COX-2) were observed. Results Compared with the A group,the B group had significantly better hemodynamics,and lower serum H-FABP,8-isoPGF2a and COX-2 levels in the early stage of post-resuscitation ( both P < 0.05 ).ConclusionsMild hypothermia attenuated post-resuscitation myocardial dysfunction during the early period of postresuscitation.The cryoprotective effect on myooardium is likely associated with the reduction of 8-iso-PGF2a and COX-2.
5.Study on the Optimization of Processing Technology of Dog Testes and Penis Processed with Talcum Pow-der and Its Quality Standard
Bin ZHAO ; Jing LIU ; Qiong WANG ; Hui CAO ; Chunsheng WEN
China Pharmacy 2016;27(19):2681-2683,2684
OBJECTIVE:To screen the optimal technology of dog testes and penis processed with talcum powder,and to pro-vide reference for the formulation of quality control standard. METHODS:Taking the comprehensive score of crushing rate and the content of alcohol soluble extract as evaluation indexes,L9(34) orthogonal test was used to optimize processing temperature,pro-cessing time and the amount of talcum powder;validation test was conducted. According to related method in Chinese Pharmaco-poeia(2015 edition),the contents of moisture,ash,nitrogen and alcohol soluble extract were determined,and automatic amino ac-id analyzer was used to determine the content of amino acids in samples. RESULTS:The optimal processing technology were that 100 kg dog testes and penis should add into 40 kg talcum powder;the processing temperature was 350-380 ℃;processing lasted for 4 min. Results of verification test showed that the average crushing rate and the content of alcohol soluble extract were 80.2%(RSD=0.95%,n=3)and 15.7%(RSD=2.30%,n=3). In processed dog testes and penis,the contents of moisture,ash,nitro-gen and alcohol soluble extract were 5.7%,18.7%,8.3%,15.7%,respectively. Besides,there still were 16 kinds of amino ac-ids,and their total content was 42.44%. CONCLUSIONS:The optimized talcum powder processing technology of dog testes and penis is stable and practical. The content of moisture,ash,nitrogen,alcohol soluble extract and amino acid can be used as impor-tant quality standard control indexes for processed dog testes and penis.
6.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.
7.Screening of High Daptomycin-producing Strain by He-Ne Laser Irradiation and Streptomycin Resistance Screening Method
Wen-Yu LU ; Jian-Ping WEN ; Jing-Hua FAN ; Bo-Xiang CAO ; Bing SUN ;
Microbiology 1992;0(03):-
The spores suspension of Streptomyces roseosporus D-38 irritated with 20mW He-Ne laser for 20 min were incubated on G1 medium plates containing 1. 9?g/mL of streptomycin. Ten percent of mutants increased the potency of daptomycin by streptomycin-resistance method, including the mutant LC-54, which could produce daptomycin 81. 2 mg/L, which was 39% higher than that of the beginning strain by flask fermentation.
8.Effects of α-methylnorepinephrine on cardiac function and myocardium at early stage of postresuscitation in rabbits
Xiaohua YANG ; Youfan FENG ; Peijie LI ; Wen CAO ; Liping ZHANG ; Jing QIN ; Xiaodong WANG
Chinese Journal of Emergency Medicine 2010;19(1):11-15
Objective To observe the effects of selective α2-adrenergic receptor agonist alpha-Methylnore-pinephrine(α-MNE) as a vasopressin agent on hemodynamics, troponin T(cTnT) and myocardium in the rabbit cardiopulmonary resuscitation. Method Eighteen health rabbits, weighing 2.5 - 3.5 kg, both male and female,were provided by Lanzhou institute of veterinary medicine. After setting up rabbit model of cardiopulmonary resuscitation, 18 rabbits were randomly divided into three groups. The rabbits in group A as a operation-control group were processed with anesthesia, endotracheal intubation, and surgery without ventricular fibrillation induced. The rabbits in group B as a epinephrine group were administered with 30 ug/kg epinephrineduring CPR. The rabbits in group C as a MNE group were administered with 100 ug/kg α-MNE during CPR. The left ventrictdar end-diastolic pressure(LVEDP), left ventricular pressure rise and fall rate(± dp/dt) and serum concentrations of BNP were measured. Statistic package of SPSS 10.0 was used for the data analysis and significant differences between means were evaluated by ANOVA analysis. Results Compared with group A, LVEDP of other two groups gradually increased respectively(P < 0. 01), and peak ± dp/dt decreased in other two groups(P<0.01). Increase in LVEDP in group C was less than that in group B(P<0.05), whereas peak ± dp/dt in group C were higher than that in group B(P<0.05), at the same stage. Compared with group A, the cTnT of the remaining two groups increased(P<0.01, respectively),and reached peak at 30 minutes. In group C, the elevation of cTnT was less than that in group B(P<0.05) during the same period. In group B and C, myocardial injury was seen under a light microscope, but the injury in group C was lighter than that in group B. Conclusion The methylnorepinephrine can lessen the myocardial dysfunction after CPR.
9.Research on contents of anthraquinones in Cassiae Semen by principal component analysis.
Li-juan CAO ; Jing MIAO ; Jie-xiu LIU ; Wen-yuan GAO ; Xia LI
China Journal of Chinese Materia Medica 2015;40(13):2589-2593
Cassiae Semen is a common traditional Chinese medicine, and contents of anthraquinones of Cassiae Semen different significantly from area to area. According to Chinese Pharmacopoeia (2010 edition), only contents of aurantio obtusin and chrysophanol were used to evaluate the quality of Cassiae Semen, another data could be added later. Ten batches of Cassiae Semen from different areas were determined, and total anthraquinones, total free anthraquinones and total combined anthraquinones contents were assessed by ultraviolet visible spectrophotometer, contents of aurantio obtusin, rhein, aloe emodin, emodin, chrysophanol and physcion were determined by HPLC. After that, principal components analysis was used to evaluate these data determined previous by dimension reduction analysis. At last, the result suggests that three main components were found out, it shows that content of aloe emodin could be used to evaluate the quality of Cassiae Semen as well as contents of aurantio obtusin and chrysophanol. And Cassiae Semen from Hebei province posseses higher quality than Cassiae Semen from other different areas. All these results can provide a good reference for quality evaluating of Cassiae Semen medicinal materials at a certain extent.
Anthraquinones
;
analysis
;
Cassia
;
chemistry
;
Chromatography, High Pressure Liquid
;
Principal Component Analysis
10.Inhibitory effect of Rhizoma typhonii extract on growth of glioma cells and its mechanism
Ling QI ; Shuang WANG ; Na WEN ; Pengyan JING ; Hong JIN ; Weiyao WANG ; Zhiyou CAO
Journal of Jilin University(Medicine Edition) 2014;(4):768-771
Objective To study the influence of Rhizoma typhonii extract on the growth of glioma SHG-44 cells cultured invitro,and to explore the mechanism of the inhibitory effect of Rhizoma typhonii extract on the growth of glioma cells.Methods The SHG-44 cells were cultured and divided into blank control group and 8, 40, 200, 1 000μg·L-1 Rhizoma typhonii extract groups.The inhibitory effect of Rhizoma typhonii extract on the growth of glioma SHG-44 cells was measured by MTT assay. The secretion levels of Bax and caspase-3 proteins were examined by ELISA assay. The expression level of caspase-3 protein was examined by Western blotting method. Results Compared with blank control group, the inhibitory rates of the growth of SHG-44 cells in 200, 1 000μg·L-1 Rhizoma typhonii extract groups at 24 h,and 8,40,200,and 1 000μg·L-1 Rhizoma typhonii extract groups at 48 h were significantly increased (P<0.05 or P<0.01).The secretion levels of Bax and caspase-3 proteins in 40,200,and 1 000μg· L-1 Rhizoma typhonii extract groups at 48 h were increased compared with blank control group (P<0.05 or P<0.01 ). Compared with blank control group, the expression levels of caspase-3 protein in different doses of Rhizoma typhonii extract groups were increased significantly (P<0.01). Conclusion Rhizoma Typhonii extract can inhibit the growth of cells through up-regulating the expression of Bax protein,increasing the expression level of caspase-3 protein and activating apoptosis pathway.