2.Effect of Peimine on ERCC1 mRNA and LRP Expressions of A549/DDP Multidrug Resistance Cell Line.
Xiao-yong TANG ; Ying-xue TANG ; Peng XU ; Hai-yan ZHOU ; Li HAN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1490-1494
OBJECTIVETo explore the effect of peimine on excision repair cross-complementation 1 (ERCC1) mRNA and lung resistant protein (LRP) expressions in A549/cisplatin (DDP) multidrug resistance (MDR) cell line.
METHODSLung cancer A549/DDP cells were cultured in vitro.Cells at logarithmic growth phase were divided into 4 groups, i.e., the blank control group, the DDP group, the ligustrazine group (DDP+ligustrazine), the peimine group (DDP + peimine). After 48-h drug action, ERCC1 mRNA expression was detected by RT-PCR and LRP expression detected by cell immunofluorescence.
RESULTSThere was no statistical difference in expression levels of ERCC1 mRNA and LRP between the DDP group and the blank control group (P > 0.05). Compared with the DDP group, expression levels of ERCC1 mRNA and LRP obviously decreased in the ligustrazine group and the peimine group (P < 0.05). They were obviously lower in the peimine group than in the ligustrazine group (P < 0.05).
CONCLUSIONSPeimine could reverse MDR of A549/DDP cell line. Its mechanism might be associated with down-regulating ERCC1 mRNA and LRP expression levels.
Cell Line, Tumor ; Cevanes ; pharmacology ; Cisplatin ; DNA-Binding Proteins ; genetics ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; Endonucleases ; genetics ; Humans ; Low Density Lipoprotein Receptor-Related Protein-1 ; genetics ; Lung Neoplasms ; RNA, Messenger ; metabolism
3.miRNA-101 inhibits the expression of the enhancer of zeste homolog 2 in androgen-independent prostate cancer LNCaP cell line.
Jian-xin LIU ; Qi-fa ZHANG ; Chang-hai TIAN ; Yong ZHANG ; Xiao-zhou HAN ; Hao GUO
National Journal of Andrology 2015;21(6):500-503
OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.
METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.
RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.
CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.
Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection
4.Analysis phylogenetic relationship of Gynostemma (Cucurbitaceae).
Shuang-shuang QIN ; Hai-tao LI ; Zhou-yong WANG ; Zhan-hu CUI ; Li-ying YU
China Journal of Chinese Materia Medica 2015;40(9):1681-1687
The sequences of ITS, matK, rbcL and psbA-trnH of 9 Gynostemma species or variety including 38 samples were compared and analyzed by molecular phylogeny method. Hemsleya macrosperma was designated as outgroup. The MP and NJ phylogenetic tree of Gynostemma was built based on ITS sequence, the results of PAUP phylogenetic analysis showed the following results: (1) The eight individuals of G. pentaphyllum var. pentaphyllum were not supported as monophyletic in the strict consensus trees and NJ trees. (2) It is suspected whether G. longipes and G. laxum should be classified as the independent species. (3)The classification of subgenus units of Gynostemma plants is supported.
Gynostemma
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classification
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genetics
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Molecular Sequence Data
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Phylogeny
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Plant Proteins
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genetics
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Sequence Analysis, DNA
5.Preparation of human meniscus acellular matrix.
Yu ZHOU ; Yu-Jie LIU ; Jing-Xiang HUANG ; Rui WANG ; Lei ZANG ; Yong HAI
China Journal of Orthopaedics and Traumatology 2014;27(3):237-239
OBJECTIVETo investigate development of a cell extraction process for preparing human meniscus acellular matrix, and morphology and biomechanical properties.
METHODSHuman meniscus were subjected to modified eight-step detergent, then, the specimens were assessed by staining with haematoxylin-eosin, toluidine blue, sirius red, saffron O, alcain blue and hoechst-33258, et al. The ultrastructure of the specimens was observed with scanning electron microscope. Transient recovery rate of deformation, maximal recovery rate of deformation and maximal compressive strength were tested to determine the biomechanical properties of the scaffold.
RESULTSEvery stain confirmed that the celluar constituents of the specimens were removed. The specimens stained positively by staining with sirius red. Lacuna were found irregularly not only on the surface of the meniscus,but also in the meniscus with scanning electron microscope. Pores in the specinmens were large, the diameter of pores was 80 to 760 microm, porosity was over 67%. The transient recovery rate of deformation was (89.62 +/- 1.04)%, the maximal recovery rate of deformation was 100% and the maximal compressive strength was (3.04 +/- 0.13)N, when the specimens were compressed 30%.
CONCLUSIONThe modified eight-step detergent can remove the immunogenic cell components from human meniscus, in addition, 3D extracellular matrix can be retained. The scaffold has good biomechanical properties. This scaffold stands a good chance to be an implant for future tissue engineering of the human meniscus.
Adult ; Cell Separation ; methods ; Cells ; chemistry ; cytology ; Cells, Cultured ; Humans ; Male ; Menisci, Tibial ; cytology ; Staining and Labeling
6.Adult extrarenal Wilms' tumor occurring in ovary: report of a case.
Li LIANG ; Xin-hua ZHOU ; Yong-jian DENG ; Hong-hai ZHANG ; Yan-qing DING
Chinese Journal of Pathology 2008;37(4):284-285
Female
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Humans
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Kidney Neoplasms
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complications
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pathology
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Ovarian Neoplasms
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pathology
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Wilms Tumor
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complications
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pathology
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Young Adult
7.Preoperative management of cardiac surgery with glucose-6-phosphate dehydrogenase deficiency
Hai-yong, WANG ; Yi-yao, JIANG ; Wen-bin, ZHANG ; Jian-fei, SONG ; Shuai-zhou, LIU
Chinese Journal of Endemiology 2011;30(6):691-693
Objective To observe the perioperative management of cardiac surgery and extracorporeal circulation method in patients with glucose-6-phosphate dehydrogenase deficiency(G6PD).Methods Ten patients with G6PD deficiency underwent uneventful cardiac surgery procedures between January 2005 and December 2010.Twenty patients who had non-G6PD deficiency were as a control group,the selected conditions were the same gender,age,body mass,the risk of heart disease surgery.The preoperative management in patients with G6PD deficiency mainly focused on avoiding the drugs implicated in haemolysis,reducing the surgical stress,using moderate hypothermia extracorporeal circulation and enhancing blood conservation.Observed indicators included the assisted ventilation time,urine volume,the drainage volume of chest tube,the amount transfusion of red blood cells and plasma,the level of hemoglobin and serum total bilirubin in the 2nd day after surgery,ICU stay.Results Compared with the control group,patients with G6PD deficiency had no significant difference in duration of ventilation after the operation,drainage,urine,Hgb,bilirubin levels,and blood transfusion[(9.3 ± 4.5)h vs (8.6 ± 5.7)h,(2100 ±670)ml vs (1950 ± 490) ml,(253 ± 146)ml vs (260 ± 120)ml,(1.3 ± 1.0)U vs (1.8 ± 1.2)U,(96 ± 25)g/L vs (99 ± 12)g/L,and (24 ± 8)μmol/L vs (27 ± 1 l)μmol/L,t =0.978,2.032,1.257,0.891,2.182,2.271,and 1.329,all P > 0.05].The duration of ICU discharge was significantly longer in the glucose-6-phosphate dehydrogenase deficient group[ (2.6 ± 0.6)d vs (1.8 ± 1.5)d,t =2.704,P < 0.05].Conclusions Cardiac surgery can be performed safely in patients with G6PD deficiency with enhanced perioperative management.
8.Significance of pupillary diameter variation in assessment of postoperative pain
Hai XIE ; Yan LI ; Duozhi WU ; Qi ZHOU ; Yong CHEN ; Naiquan MA
The Journal of Clinical Anesthesiology 2016;(2):134-138
Objective To observe the reactivity of pupillary diameter variation responding to numerical rating scale on postoperative pain to explore the accuracy of pupillary diameter variation for pain assessment.Methods Eighty patients after selected surgery (male 43 cases,female 37 cases,aged 24-79 years,ASA gradeⅠorⅡ)were included for the trial.After endotracheal extubation and on arri-val in the post-anaesthesia care unit (PACU),the levels of pain on numerical rating scale (NRS)were rated and recorded subsequently.If NRS was within 0-3,no analgesic was administered and 50 μg fentanyl was administrated after 5 min if NRS rated in 4-10.The pupillary diameter (PD),systolic blood pressure (SBP)and heart rate (HR)were recorded every minute during 20 minute after first NRS evaluation,the corresponding maximum variation of above index were calculated simultaneously in 10 minutes with NRS 0-3,after 5 minutes with NRS 0-3 to 4-10,after 5 minutes with NRS 4-10 to 0-3,in 10 minutes with NRS 4-10 which were marked respectively with ΔPD,ΔSBP and ΔHR.With the variation of NRS as a criterion,the reactivity of ΔHR,ΔSBP and ΔPD responding to NRS varia-tion were assessed by analysing the receiver operating characteristic curve (ROC),the area under the receiver operating characteristic curve (AUC)was calculated.Results The patients' ROC value ofΔHR,ΔSBP and ΔPD responding to NRS variation were AUCΔPD 0.904 (95% CI 0.822-0.987 ), AUCΔSBP 0.65 1(95%CI 0.5 10-0.781)and AUCΔHR 0.588(95%CI 0.444-0.733)respectively.Value of AUCΔPD was larger than that of AUCΔSBP and AUCΔHR respectively (P < 0.05 ).Importantly, operating characteristic curve (ROC)analysis showed that the diagnostic cutoff value of ΔPD was 41.3% with a sensitivity of 80.0% and a specificity of 93.5% and the diagnostic cutoff value of ΔSBP was 10.3% with a sensitivity of 62.4% and a specificity of 71.5%.Conclusion This study demon-strated that the accuracy of ΔPD was higher than ΔSBP and ΔHR responding to pain/analgesia evalu-ation.ΔPD was a valuable index for assessment of postoperative pain.
9.Effects of protein kinase-induced cytoplasmic calcium changes in mesangial cells on glomerular hy- perfiltration in early stage of rat diabetes mellitus
Song-Min HUANG ; Hai-Yan ZHANG ; Yong XU ; Zhao-Hui SHA ; Li ZHOU ;
Chinese Journal of Diabetes 1994;0(02):-
Objective To elucidate functional alterations of glomerular mesangial cell (GMC) contraction and its relationship with diabetic glomerular hyperfiltration. Methods Male Wistar rats were divided into groups of normal control and 2 weeks diabetes induced by STZ. GMC were cultured in the presence of normal glucose (5mM). Meanwhile in comparison GMC obstained from normal rats were cultured in the presence of 30 mM glucose for 48 h in vitro. By using laser scanning confocal microscopy (LSCM) , we measured basal calcium concentration, the changes of calcium level in response to vasoconstrictor agonist angiotensin H (Ang I ),and the effect of protein kinase C (PKC) inhibitor PMAC10 M,24 h)on cytosol calcium of GMCs. Results The creatinine clearance rate (Ccr) was increased with age in diabetic rats, reaching a peak at 2 weeks (P