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.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.
5.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
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.One-stage repair of congenital aortic arch disease with other cardiac defects by using autologous pulmonary artery tissue.
Hui XUE ; Qing-yu WU ; Hong-yin LI ; Ming-kui ZHANG ; Ji-cheng XI ; Guang-yu PAN ; Zhong-hua XU
Chinese Journal of Surgery 2010;48(10):724-726
OBJECTIVETo analyze the surgical strategy and result of one-stage repair for congenital aortic arch disease associated with other cardiac anomalies.
METHODSBetween April 1993 and November 2009, 25 consecutive patients aged 26 d to 6.5 years underwent one-stage repair for congenital aortic arch disease with other cardiac anomalies. Among them, 6 patients had coarctation of aorta, 6 patients had interrupted aortic arch, and 13 cases had hypoplasia of aortic arch. The surgical techniques include excision of the anterior wall of pulmonary artery, resection of patent ductus arteriosus tissue, aortic arch reconstruction with autologous pulmonary artery wall, reconstruction of the pulmonary artery and repair of the associated defects.
RESULTSTwenty-four patients survived and recovered uneventfully. One patient died of pulmonary hypertension crisis in hospital. The reconstruction of the aorta and the correction of the intracardiac anomalies were proved by postoperative echocardiography and CT scan. There were no neurological or other complications. The follow-up showed that all patients developed normally and there were no restenosis of the aorta arch.
CONCLUSIONSWith the benefits of growth potential and less tension, autologous pulmonary artery tissue is an optimal choice in aortic arch reconstruction. One-stage repair of congenital aortic arch disease associated with other cardiac anomalies can achieve good results.
Aorta, Thoracic ; abnormalities ; surgery ; Aortic Coarctation ; complications ; surgery ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; complications ; surgery ; Humans ; Infant ; Infant, Newborn ; Male ; Pulmonary Artery ; transplantation ; Retrospective Studies ; Transplantation, Autologous ; Treatment Outcome
9.Clinical significance of protein Z alteration in patients with cardio-cerebral thrombotic diseases.
Xue-yi PAN ; Cai-ping DING ; Liang-yi ZHONG ; Xu-ming HUANG ; Wan-xing ZHOU ; Yu GUO ; Jin-zhu YIN ; Xiao-yan CAI ; Ze-bing GUAN ; Rou-ling ZHANG
Chinese Journal of Hematology 2004;25(11):671-674
OBJECTIVETo study the alteration of protein Z (PZ) in patients with cardio-cerebral thrombotic diseases, its clinical significance and relations with FX.
METHODSPZ and FX:Ag were measured by ELISA, and plasma FX:C by first stage method. In 170 patients with acute ischemic stroke (AIS), 40 acute myocardial infarction (AMI) and 60 healthy adults as contrast, PZ, FX:C and FX:Ag were measured and compared between incipience and recurrence, different ages and genders.
RESULTSIn AIS and AMI groups, PZ levels decreased significantly to (940.02 +/- 229.82) microg/L and (1071.44 +/- 180.52) microg/L, respectively \[the contrast group was (2257.97 +/- 479.76) microg/L, P < 0.001\]. But FX:C and FX:Ag raised to (136.73 +/- 34.93)% and (135.54 +/- 54.39)% in AIS group; and to (139.53 +/- 29.18)%, (129.75 +/- 21.91)% in AMI group, respectively, while in the contrast group they were (94.33 +/- 22.00)% and (77.22 +/- 13.19)% (P < 0.001). In the comparative research between the AIS group, AMI group and the contrast group, PZ level was clearly found to negatively relate to the level of FX:C and FX:Ag (P < 0.001). Meanwhile, PZ level, FX:C and FX:Ag in recur-AIS group and recur-AMI group exhibited significant differences (P < 0.05) from those in the primary AIS and AMI groups, suggesting that the decrease of PZ levels reflected the pathological process of the disease. In addition, PZ level gradually decreased with the increase of age (P < 0.05), while FX:C and FX:Ag had no relations with age (P > 0.05). No correlation was found in sex with PZ level, FX:C, FX:Ag (P > 0.05).
CONCLUSIONPZ level was significantly decreased in AIS and AMI patients and was negatively related to FX:C and FX:Ag. The mechanism leading to FX increase may partially related with the decreased of PZ. PZ level was different in the primary and recurrent disease and was gradually decreased with the increase of age. Lack of PZ might be a etiological factor of cardio-cerebral thrombotic diseases.
Aged ; Aged, 80 and over ; Blood Proteins ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Factor X ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; Stroke ; blood
10.Differential expressions of lipid metabolism related genes in the liver of young apoE knockout mice.
Hong-Yan YE ; Miao YIN ; Yun-Ju SHANG ; Xue-Dong DAI ; Sheng-Qiang ZHANG ; Wen JING ; Hui-Qin DU ; Liang ZHANG ; Jie PAN
Acta Physiologica Sinica 2008;60(1):51-58
The work was aimed to investigate the differential expressions of lipid metabolism related genes in the early stage of atherosclerosis in the young apolipoprotein E deficient (apoE(-/-)) mice at different ages with normal chow diet. The genotypes of mice were identified by using multiplex polymerase chain reaction (multi-PCR) analysis. The semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR were used to analyze the expressions of lipid metabolism related genes in the liver of apoE(-/-) and age-matched wild type (WT) mice of 14-day old, 1-month old, 2-month old, 3-month old. The serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) contents were assayed using COD-PAP and GPO-PAP methods. The serum apolipoprotein B100 (apoB100) content was quantitated by immune turbidimetry. The hearts were perfusion-fixed in 4% formaldehyde, infiltrated with 30% gum sucrose for 24 h at 4 °C, and embedded in OCT compound. The aortic sinus tissues were serially sectioned at -15 °C, stained with Sudan IV, and counterstained with light green. The results were shown as follows. Compared with that in WT mice, the mRNA levels of apoA I and apoA IV in apoE(-/-) mice aged from 14-day old to 3-month old changed prominently (P<0.05), with apoA I up-regulated and apoA IV down-regulated. At the age of 1 month, the expression of apoB100 in apoE(-/-) mice was higher than that in WT mice (P<0.05). The expression of apoA V was up-regulated (P<0.05) and there was obvious lipid deposition in the aortic intima in apoE(-/-) mice at the age of 2 months. The expressions of fatty acid translocase (Fat/CD36) and angiopoietin-like protein 3 (Angptl 3) in apoE(-/-) mice were higher than those in WT mice at the age of 3 months (P<0.05), while the expressions of peroxisome proliferator-activated receptor α (PPARα), liver X receptor α (LXRα), carnitine palmitoyl transferase I (CPT I) and acyl coenzyme A oxidase 1 (ACOX1) showed no significant changes. The serum TC, TG, LDL-C and HDL-C contents in apoE(-/-) mice aged from 14-day old to 3-month old were higher than those in age-matched WT mice. apoE(-/-) mice showed a marked increase in serum apoB100 content, consistent with the trend of serum LDL-C content and apoB100 mRNA content in the liver. The results suggest that the mRNA expressions of apoA I, apoA IV, apoA V, apoB100 and Angptl 3 in apoE(-/-) mice change significantly compared with those in WT mice, and these genes might be relevant to the complicated lipid metabolism network, and involved in the early stage of atherogenesis.
Animals
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Apolipoprotein A-I
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metabolism
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Apolipoprotein B-100
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blood
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Apolipoproteins A
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metabolism
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Apolipoproteins E
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genetics
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Atherosclerosis
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genetics
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Gene Expression
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Lipid Metabolism
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genetics
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Lipoproteins, HDL
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blood
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Lipoproteins, LDL
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blood
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Liver
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metabolism
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Mice
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Mice, Knockout
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Triglycerides
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blood