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 Effect of Quercetin Targeted p53 Gene and Protein Expression in NB4 Cell Lines
Qing-wen, XIE ; Jing-qiu, ZHAO ; Zhi-wen, FANG
Journal of Shanghai Jiaotong University(Medical Science) 2001;21(1):8-10
Objective In order to study the value of drug treatment for leukemia, quercetin was delivered to the cultured NB4 cell line. MethodsUsing RT- PCR and western blot, we studied the expression of mutant p53 gene and protein after inducing 24, 48, 72, 96 and 120h by the quercetin with different concentrations (30, 60, 90μmol/L). Results We verified the results that quercetin with concentration of 30 ~90μmol/L could successfully inhibit the expression of mutant p53 protein, but the mutant p53 gene did not have. ConclusionThat data suggested that quercetin would be an effective method for the therapy of leukemia.
3.Study on the Quality Standard of Liyan Mixture
Xia JING ; Jin XU ; Fang SUN ; Wen ZHAN
China Pharmacy 2015;26(36):5141-5143
OBJECTIVE:To establish the quality standard of Liyan mixture. METHODS:TLC was conducted to identify the Scrophularia ningpoensis,Terminalia chebula and Glycyrrhiza uralensis;HPLC was conducted to determine the content of chloro-genic acid. The column was Agilent Eclipse Plus C18 with mobile phase of methanol-0.2% phosphoric acid (18:82,V/V) at flow rate of 1.0 ml/min,the detection wavelength was 327 nm,column temperature was 30 ℃ and the injection volume was 20 μl. RE-SULTS:TLC showed S. ningpoensis,T. chebula and G. uralensis had clear spots and good separation. There was no interference for negative control. The linear range of chlorogenic acid was 0.16-1.6 μg(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were lower than 1.0%,recovery was 96.9%-101.4%(RSD=1.17%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Liyan mixture.
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.
6.Progress in management of patent ductus arterious in preterm infants with gestational age of <28 weeks.
Chinese Journal of Contemporary Pediatrics 2015;17(10):1142-1147
Patent ductus arteriosus (PDA) is a common problem encountered in the early neonatal period, particularly in preterm infants. Optimal management of PDA in preterm infants remains controversial. Despite considerable historical and physiological data indicating a persistent PDA may be harmful, robust evidence of long-term benefits or harms from treatment is lacking. This has been equated to a lack of benefit but is also a reflection of the fact that many clinical trials were designed to assess the effects of short-term (2-8 days) rather than prolonged exposure to a PDA. No clinical trials have been designed to assess the effects of prolonged exposure of persistent PDA on morbidity and mortality of very premature infants. Significant changes in management of PDA, i.e., less treatment for PDA, have evolved in recent years. This paper reviews the current literature and evidence for treatment options and research progress of PDA in infants with gestational age of <28 weeks.
Ductus Arteriosus, Patent
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therapy
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Gestational Age
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Humans
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Infant, Newborn
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Infant, Premature
8.Novel drug composition ameliorating thrombosis and its molecular mechanisms.
Meng NING ; Jing-Hui HUANG ; Yan-Fang ZHANG ; Wen-Yu CUI ; Hai WANG
Chinese Journal of Applied Physiology 2014;30(2):184-188
OBJECTIVETo investigate the antithrombotic effects and its molecular mechanisms of prazosin combined with anisodamine (Ani).
METHODSIsolated rat tail artery rings model was employed to evaluate the vasodilative effects of drugs, mice tail thrombosis model induced by carrageenan was used to study the antithrombotic effects and its molecular mechanisms of the drug composition.
RESULTSAmong alpha1-adrenoreceptor antagonists, prazosin(Pra) had the greatest relaxation rate, which was (82.6 +/- 8.9)%, and the EC50 value was 0.44 micromol/L. The drug composition of anisodamine and prazosin of different doses could decrease the length of the tail thrombosis from (24.6 +/- 4.6)mm to (6.9 +/- 2.7)mm, and the rate of thrombosis was decreased from 86.6% to 50.0%. The drug composition could prolong the prothrombin time (PT) distinctively, but it had no effect on the activated partial thromboplastin time (APTT). It also could restrain the decrease of serum levels of tissue plasminogen activator (t-PA) and 6- Keto -PGF1alpha as well as the increase of type-1 plasminogen activator inhibitor (PAI-1) and thromboxane B2 (TXB2) in the mice.
CONCLUSIONThe drug composition formed by anisodamine and prazosin has good effects of relaxing extremities tiny blood vessels and it can fight against thrombosis, its antithrombotic mechanisms may be related to the influence of the extrinsic coagulation pathway, inhibition of platelet activation functions and the promotion of fibrinolysis function.
Adrenergic alpha-1 Receptor Antagonists ; pharmacology ; Animals ; Disease Models, Animal ; In Vitro Techniques ; Male ; Prazosin ; pharmacology ; Rats ; Rats, Wistar ; Solanaceous Alkaloids ; pharmacology ; Thrombosis ; drug therapy
9.Weil osteotomy for metastatic metatarsalgia after hallux valgus surgery
Jing LI ; Kun LI ; Ming XIE ; Jianmin WEN ; Zhenhua FANG ; Ruokun HUANG ; Jingjing ZHAO
Chinese Journal of Orthopaedics 2014;(10):1024-1029
Objective To investigate the cause of metastatic metatarsalgia after hallux valgus surgery and the clinical outcomes of Weil osteotomy for metastatic metatarsalgia after hallux valgus surgery. Methods From July 2009 to Janurary 2012, data of 27 patients (27 feet) with metastatic metatarsalgia of 2nd to 4th head of metatarsal bone after hallux valgus surgery who had been treated by Weil osteotomy were retrospectively analyzed. There were 1 male and 26 females with an average age of 51 years (range, 28-73 years). Metatarsalgia occurred 6-24 months after operation for hallux valgus. 13 feet underwent mini-invasive cervi-cal wedge osteotomy of the first metatarsal, 7 with Chevron procedure, 5 with Akin procedure, and 2 with Lapidus procedure. No shortening in first metatarsal was found in 5 feet with Akin osteotomy, while there were varying degrees of shortening in first meta-tarsal in the remaining 22 feet. The clinical results were evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) and visual analogue scale (VAS) scores. The preoperative and postoperative plantar pressure changes were tested by plantar pressure tester. Results 25 Patients were followed up for 12-42 months (average, 24 months). Among these 25 cases, the metatarsalgia of 23 cases were completely disappeared. The metatarsal plantar lateral metastatic pain occurred in the remaining 2 cases (2 feet) and 1 was relieved by the foot pad, 1 was cured by re-Weil osteotomy. AOFAS score was 46.82 ± 6.13 before surgery and 90.63 ± 1.65 after surgery. The VAS score was 7.5 (6, 7) before surgery and 0.5 (0, 1.0) after surgery. The last follow-up, according to the score of AOFAS toe metatarsophalangeal-interphalangeal joint, of which 23 cases were excellent, 1 good, 1 poor;the excellent and good rate was 96%(24/25). Preoperative pressure under 2nd to 5th metatarsal head were 3.12±1.62 Pa, 5.81±1.92 Pa, 4.63± 2.10 Pa, 3.37±1.57 Pa, 1.67±1.20 Pa and postoperative were 3.33±1.35 Pa, 3.89±1.08 Pa, 3.65±1.96 Pa, 2.25±1.23, (1.48±1.11) Pa. Postoperative pressure under 2nd to 5th metatarsal head were significantly decreased. Conclusion Weil osteotomy can effec-tively adjust the length of the metatarsal and the height of metatarsal head, thus effectively improve the pressure under the metatar-sal head, so it could reach a good effect in the treatment of metastatic metatarsalgia after hallux valgus surgery.
10.Analysis of CUMS-induced differentially expressed proteins in rat hippocampus by ITRAQ
Lisha CAO ; Fang ZHANG ; Wen LUO ; Jing CHEN ; Miaoquan HE ; Jisheng WANG
Chinese Pharmacological Bulletin 2016;32(5):697-702
Aims To compare hippocampus between CUMS rats and normal rats, to find differentially ex-pressed proteins and to explore the pathogenesis of de-pression in the protein levels and biological marker. Methods Chronic unpredicted mild stress was taken to establish rat depression model. The ITRAQ-labeled proteins and peptides were separated by the cation col-umn, and differentially expressed proteins were detec-ted and identified by 2D LC-MS/MS. The functions of proteins were analyzed by bioinformatics. Results To-tally 5 109 proteins were identified, 33 differentially expressed proteins were identified, the expressions of 8 proteins were increased and 25 proteins downregulated. Conclusion ITRAQ based sereening is effective in discovering the nosogenesis of depression and new bio-logical marker.