1.A clinical and laboratory investigation of acute promyelocytic leukemia with tetraploid clone characterized by two t(15 ;17)
Yisun FAN ; Xia SHEN ; Jinlan PAN ; Yongquan XUE ; Xufang YIN
Chinese Journal of Laboratory Medicine 2010;33(6):522-526
Objective To investigate clinical and experimental features of APL with tetraploid clone characterized by double t (15 ; 17). MethodsFive cases of APL with tetraploid clone characterized by double t(15;17) were chosen. Cytogenetic examination of bone marrow was performed with bone marrow or short-period culture. R banding technique was used for karyotype analysis. DNA content in one case was determined by flow cytometry. Immunophenotyping was performed by using a panel of monoclonal antibodies :CD2, CD13, CD15, CD33 and CD34. PML/RARα fusion gene was detected by interphase FISH using dualcolor PML/RARα probe in one case and by RT-PCR in two cases. ResultsAll cases were male with a median age of 38. Their marrow cell morphology examination showed marked hyperplasia with large leukemic cells that had bizarre nuclear configuration. Chromosome analysis revealed that a leukemia clone with tetraploid or near-tetraploid karyotype characterized by double t(15;17) (q22 ;q12) in five cases, of which, one also had a diploid clone with t(15;17) and a normal cell;two had some cells with normal karyotypes.PML/RARα fusion gene was detected by FISH in one of 5 cases and by RT-PCR in 2 of 5 cases. CD33 expression was found in one case. CD13 and CD33 expressions were seen in the other four cases, of which,CD34 or CD2 co-expression was found in one case and in two cases respectively. The result of DNA content showed a single peak which indicated only tetraploid clone whose DNA index was 1. 998 with CV of 8. 2%.All patients obtained complete remission after the treatment with ATRA and/or arsenic trioxide. Conclusions These results indicate that API, patients with tetraploidy and near-tetraploidy have giant and bizarre blasts.Most patients have short-type PML/RARα transcripts. The tetraploidy in APL does not appear to affect the response to treatment of ATRA.
2.Establishment of a Hybridoma Cell Line Secreting High Titer of Anti-Human Thyroglobulin Monoclonal Antibodies
Yanhua YAN ; Enjiang TIAN ; Shuyou CHONG ; Yin XUE ; Xiaoxi HU ; Jufen PAN ; Wufei ZHENG
Chinese Journal of Immunology 1985;0(05):-
A hybridoma cell line(B10)was established by fusing spleen cells of BALB/c mou- se immunized with human thyroglobulin(HTG) with SP2/0 myeloma cells. An averagefusing rate of 6.3% and antibody-secreting positive well rate of 58.3% were obtained.During the first two months, the supernatant of B10 culture had a titer of 1/128 to1/2048 measured by hemagglutination method, and the ascites was positive at 1/64000-128000 and 1/320000 respectively as measured by hemagglutination and radioimmunoass-ay.The B10 cell line is very stable and has very high activity to produce anti-HTGmonoclonal antibodies. After several times of preservation in liquid nitrogen andpassage in culture for one year,a recent determination shows that cell culture super-natant and ascites still have very high titer,being 1/4096 and 1/1048576 respectively asmeasured by hemagglutination method. The chromosome number of the B10 hybridomacell is 99.5?7.4.The success of the establishment of this cell line is briefly discussed.Attempt to establish diagnostic kit with this monoclonal antibody is being undertaken.
3.Effect of L-arginine on expression of PKC mRNA in pulmonary injury induced by ischemia-reperfusion in rabbits
Wan-Tie WANG ; Fang-Yan WANG ; Shou-Quan CHEN ; Yin-Fei YU ; Xue-Rong PAN ; Xi-Wen CHEN ;
Chinese Journal of Emergency Medicine 2006;0(12):-
Objective To investigate the effect of L-arginine on expression of protein kinase C(PKC)mRNA during pulmonary ischemia and reperfusion injury(PIRI)in the rabbits.Methods Single lung ischemia and reperfusion animal model was used in vivo.The rabbits were randomly divided into three groups(n=9,in each),sham operated group (Sham),PIR group(I-R)and PIR+L-arginine group(L-Arg).Changes of several rariables including malondialdehyde (MDA),superoxide dismutase(SOD),malandialde hyde(MDA),nitril oxide(NO),wet to dry ratio of lung tissue weight(W/D)and index of quantitative assessment(IQA)of histolngic lung injury were recorded at 60 minutes after reperfusion in lung tissue.Meanwhile the location and expression of PKC mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60 minutes after reperfusion.Results In comparison with group I-R,PKC mRNA strongly expressed in intima and extima of small pulmonary artery as well as thin-waU vessels (mostly small pulmonary veins).The average optical density values of PKC-?,?and?mRNA in small pulmonary veins in L-Arg group had significance(all P<0.01);SOD increased while MDA,W/D and IQA decreased at 60 minutes after reperfusion in lung tissue(P<0.01 and P<0.05).A morphologically abnormal changes of the lung tissue,were lessen markedly in L-Arg group.Conclusion L-arginine possess notably protective effects on PIRI in rabbits by activating PKC-?,?and?mRNA expression in lung tissue,raising NO level,dropping oxygen free radical level and decreasing lipid peroxidation.
4.The interference in correlated molecular mechanism obtained multi-drug resistance of mouse S180's tumour cell for different alkaloid.
Gui-hai LI ; Cheng-ye PAN ; Fu-jun SUN ; Ge-ping YIN ; Xue-rong WANG
China Journal of Chinese Materia Medica 2005;30(23):1844-1848
OBJECTIVETo observe the base of the interference in correlated biotic active matter obtained multi-drug resistance induced by chemotherapy for different alkaloid, and to supervise the use in clinic to restrain the multi-drug resistant of chemotherapy, and thereby to improve the curative effect.
METHODAfter bestowing subter-dosage unite chemotherapeutant to ascites S180 mouse to set up the mouse models of multi-drug resistance of S180 tumour cell, and giving the mouse matrine, terandrine, oxymatrine and berberine hydrooh loride for 4 weeks, the P170, LRP, TOPOII, Fas and apoposis were determined by flow cytometry.
RESULTMatrine and terandrine could obviously reduce the express of P170, LRP and the activation of TOPOII, and increase the ratio of the express of Fas and the apoposis of drug resistant tumour cell. And at the same time it could obviously reduce the express of intercellular adhesion molecule(CD54).
CONCLUSIONMatrine and terandrine can interfere in MDR which results from chemotherapeutics by the adjustment of correlated biotic active matter, besides, the different degree of alkaloid effect with different configuration.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Alkaloids ; isolation & purification ; pharmacology ; Animals ; Apoptosis ; drug effects ; Benzylisoquinolines ; isolation & purification ; pharmacology ; Berberine Alkaloids ; isolation & purification ; pharmacology ; DNA Topoisomerases, Type II ; metabolism ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Male ; Mice ; Plants, Medicinal ; chemistry ; Quinolizines ; isolation & purification ; pharmacology ; Random Allocation ; Sarcoma 180 ; metabolism ; pathology ; Tumor Cells, Cultured ; Vault Ribonucleoprotein Particles ; metabolism ; fas Receptor ; metabolism
5.p53 Anti-tumor Research in Bel-7402 by Using Human-derived Vector
Zhigang XUE ; Jian LI ; Biao YIN ; Yakun ZHANG ; Xionghao LIU ; Qian PAN ; Zhigao LONG ; Heping DAI ; Kun XIA ; Lingqian WU ; Desheng LIANG ; Jiahui XIA
Progress in Biochemistry and Biophysics 2007;34(5):465-470
In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.
6.Value of plasma miR-193a-5p level on diagnosis and treatment evaluation in acute myeloid leukemia
Na ZHANG ; Zhifang XU ; Fanggang REN ; Junxia ZHAO ; Jing XU ; Xiuhua CHEN ; Yanhong TAN ; Jianmei CHANG ; Feng XUE ; Feng GAO ; Jie PAN ; Bin YIN ; Hongwei WANG
Journal of Leukemia & Lymphoma 2016;25(6):349-353
Objective To explore the value of the plasma miR-193a-5p level on diagnosis and monitoring the response to treatment in acute myeloid leukemia (AML). Methods Peripheral blood samples were obtained from AML patients enrolled in hematology department of the Second Hospital of Shanxi Medical University from July 2015 to December 2015, including 30 de novo AML patients, 9 patients in complete remission (CR) and 6 patients in relapse. Peripheral blood samples from 15 healthy people were randomly choosed as the health control group. Plasma miR-193a-5p expression levels were detected by using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results The plasma miR-193a-5p relative expression level of AML patients group [0.465 6 (0.103 1-5.000 2)] was obviously lower than that of health control group [0.766 1 (0.052 1-3.134 4)] (U= 122, P= 0.018 7). The plasma miR-193a-5p relative expression levels of de novo group and relapse AML group were significantly lower than those of CR group and health control group (P<0.05), and there was no significant difference between the CR group and health control group (U= 56, P= 0.511 9). No significant correlation was found between the plasma miR-193a-5p level and age, gender, blast percentage of the bone marrow, peripheral blood leukocyte count, platelet count, CD34+cells'percentage and so on. Conclusion The decreased plasma miR-193a-5p expression level can be served as a new and noninvasive biomarker for the evaluation of diagnosis and treatment in AML.
7.Influence of adefovir dipivoxil on HBV specific CTL in patients with chronic hepatitis B
Yu-Lin ZHOU ; Xue-Cai WANG ; Yin-Tao WU ; Yong-Fei TAN ; Yan-Ping ZHAO ; Jun-Ming TANG ; Jian-Qiang PAN
Chinese Journal of Experimental and Clinical Virology 2010;24(5):362-363
Objective To explore the influence of adefovir dipivoxil on HBV specific CTL in patients with chronic hepatitis B (CHB). Methods 10 mg adefovir dipivoxil (Zhengda Tianjing Pharmaceutical Company) was used for CHB patients with positive HBV DNA (HBV DNA ≥ 1 × 104 copies/ml) ,ALT > 2 × upper limit of normal value (ULN) and positive human leucocyte antigen (HLA)-A2,orally,once a day for 3 months. Real time fluorescent quantitative PCR was used to determine HBV DNA and flowcytometer was used to determine HBV specific CTL. Results After treatment with adefovir dipivoxil for 3 months, HBV specific CTL (0. 52 ± 0. 11 )% was higher than that before treatment(0. 34 ± 0. 14 )% , t =6.78 P <0. 01, HBV DNA of 28 cases turned to negative ( < 1 × 103copies/ml) (62. 22% ). HBV DNA of 17 cases failed to turn negative 3 months after treatment,but their HBV DNA level was lower [(4. 18 ±0.4)log 10 copies/ml] than that before treatment [(6. 23 ± 0. 73 ) log 10 copies/ml], t = 9.99, P < 0.01.Conclusion adefovir dipivoxil can improve HBV specific cellular immunity in patients CHB.
8.Study on the differential expression of lipid metabolism-related genes in young LDLR knockout mice liver.
Yun-Ju SHANG ; Xue-Dong DAI ; Wen JING ; Hui-Qin DU ; Hong-Yan YE ; Miao YIN ; Liang ZHANG ; Sheng-Qiang ZHANG ; Ji-Feng LI ; Jie PAN
Chinese Journal of Pathology 2008;37(3):179-183
OBJECTIVETo clarify the differential expression of the genes related to the lipid metabolism in the early stage of atherosclerosis in the young LDLR-/- mice of different ages.
METHODSA RT-PCR assay was used to analyse the gene expression patterns in the livers of LDLR-/- mice and wild type (WT) mice from 14 to 90 days. The characteristics of early lipid deposition in intima were evaluated using biochemical and pathological techniques.
RESULTSIn LDLR-/- mice, when compared to WT mice, the mRNA level of the apolipoprotein A IV (apoA IV), fatty acid translocase (Fat/CD36) and carnitine palmitoyl transferase I (CPT I) changed prominently at the age of 14-days (P < 0.05). At 30 days, the mRNA level of apolipoprotein A I (apoA I) was up regulated, but apolipoprotein F (apoF), CD36 and CPT I were down regulated (P < 0.05). At 60 days, the mRNA levels of apoA I, CPT I and liver X receptor alpha (LXRalpha) were up regulated, but apoA IV was down regulated (P < 0.05). At 90 days, the level of the apoA I was higher, but the expression of the apoA IV, apoF and acyl-coenzymeA oxidase 1 (ACOX1) were down regulated (P < 0.05), whereas the expression of apolipoprotein A V (apoA V), apolipoprotein E (apoE), peroxidase proliferator-activated receptor alpha (PPARalpha) and angiopoietin-like protein 3 (angptl 3) had no significant changes (P > 0.05). The serum levels of TC (P < 0.05), TG (P < 0.05) and LDLC (P < 0.05) in LDLR-/- mice were significantly higher than those in wild type mice with the same age.
CONCLUSIONSThe mRNA levels of the apoA I, apoA IV, apoF, FAT/CD36, CPT I, ACOX1 and LXRalpha of the LDLR-/- mice were significantly changed compared to the WT mice. The genes may be of some relevance to the complicated lipid metabolism network, and have effect in the early stage of atherogenesis.
Animals ; Apolipoprotein A-I ; genetics ; metabolism ; Apolipoproteins A ; genetics ; metabolism ; Apolipoproteins E ; genetics ; metabolism ; Gene Expression ; Lipid Metabolism ; Liver ; metabolism ; Liver X Receptors ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Orphan Nuclear Receptors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Receptors, LDL ; deficiency
9.Expression profiles of lipid metabolism-related genes in liver of apoE(-/-)/LDLR(-/-) mice.
Hui-qin DU ; Miao YIN ; Hong-yan YE ; Yun-ju SHANG ; Xue-dong DAI ; Wen JING ; Liang ZHANG ; Ning XIAO ; Ji-feng LI ; Jie PAN
Chinese Journal of Pathology 2007;36(11):751-755
OBJECTIVETo explore the relationship between the expression characteristics of lipid metabolism-related genes in the liver and early atherosclerotic lesions in apolipoprotein E and low density lipoprotein receptor gene double knockout (apoE(-/-)/LDLR(-/-)) mice.
METHODSRT-PCR was used to detect the differential expression of lipid metabolism-related genes in the liver of apoE(-/-)/LDLR(-/-) and wild type (WT) mice. Serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) level as well as aortic morphology were also analyzed.
RESULTSAmong the 11 lipid metabolism-related genes, apolipoprotein B100 (apoB100) mRNA levels were significantly higher in apoE(-/-)/LDLR(-/-)mice compared with WT mice. At 14 days, 1, 2 and 3 months of age, the level of mRNA expression were 1.55, 1.47, 1.50 and 2.42 folds of those of the age matched WT mice respectively. The fatty acid transporter (FAT/CD36) mRNA expression levels were higher in 14-day and 3-month old mice at 1.30 and 1.35 folds of those of the age matched WT mice, respectively. Apolipoprotein A IV (apoA IV) and Apolipoprotein AV (apoAV) mRNA levels were significantly down-regulated (0.89 fold decrease in 14-day, and 0.90 folds decrease in 3-month, respectively). The mRNA expression levels of apolipoprotein AI (apo AI), apolipoprotein F (apo F), peroxidase proliferator-activated receptor alpha (PPAR-alpha), liver X receptor alpha (LXRalpha), angiopoietin-like protein 3 (ANGPTL3), acyl-coenzymeA oxidase 1 (ACOX1) and carnitine palmitoyl transferase 1 (CPT1) had no significant changes. Serum TC, TG and LDL-C were higher than those of age matched WT mice at 7, 2 and 30 folds, respectively. Furthermore, apoE(-/-)/LDLR(-/-) mice demonstrated typical early atherosclerotic lesions at sinus and root regions of aorta in an age dependent manner.
CONCLUSIONAlterations of the expression of lipid metabolism-related genes in liver play important roles in the development of AS in the apoE(-/-)/LDLR(-/-) mice at early ages.
Animals ; Aorta ; pathology ; Apolipoprotein A-V ; Apolipoprotein B-100 ; biosynthesis ; genetics ; Apolipoproteins ; biosynthesis ; genetics ; Apolipoproteins A ; biosynthesis ; genetics ; Apolipoproteins E ; deficiency ; Atherosclerosis ; etiology ; metabolism ; pathology ; CD36 Antigens ; biosynthesis ; genetics ; Gene Expression ; Lipid Metabolism ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; RNA, Messenger ; metabolism ; Receptors, LDL ; deficiency
10.Surgical treatment of complex congenital heart diseases with extracardiac conduit total cavopulmonary connection.
Qing-yu WU ; Hong-yin LI ; Ming-kui ZHANG ; Xing-peng CHEN ; Guang-yu PAN ; Ji-cheng XI ; Hui XUE
Chinese Journal of Surgery 2007;45(12):805-807
OBJECTIVETo report the experience of extracardiac conduit total cavopulmonary connection (ECTCPC) in surgical treatment of complex congenital heart diseases.
METHODSFrom 1998 to 2006, 68 patients underwent ECTCPC for complex congenital heart diseases. Among them, 45 had functional univentricle with transposition of the great artery (TGA) and pulmonary artery valve stenosis, 19 had tricuspid atresia with hypoplasia of right ventricle, 4 had Ebstein's anomaly with hypoplasia of right ventricle. Six had left superior vena cava, 18 had received Bidirectional Glenn operation; Fifty-seven cases were performed under cardiopulmonary bypass with general anesthesia and hypothermia, 11 cases were performed without cardiopulmonary bypass.
RESULTSThere were two death, the mortality was 2.9%. All patients were followed up from 1 to 8 years with no clinical symptoms and have been doing well. The arterial oxygen saturation was 90% - 96%, the cardiac function were in NYHA class I - II.
CONCLUSIONThe extra cardiac conduit TCPC is a simple procedure and superior to other type of Fontan procedure in most patients.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Fontan Procedure ; methods ; Heart Defects, Congenital ; surgery ; Humans ; Male ; Treatment Outcome