Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 15 de 15
Filter
1.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 144-149, 2020.
Article in Chinese | WPRIM | ID: wpr-787611

ABSTRACT

Meta-analysis was used to compare the long-term efficacy and laryngeal function preservation rate of patients with advanced hypopharyngeal cancer treated with surgery plus radio(chemo)therapy (SRT) or non-surgery chemoradiotherapy (CRT). We searched publicly published articles on case-control studies of surgical and non-surgical comprehensive treatment of advanced hypopharyngeal cancer in PubMed, the Cochrane Library, Wanfang Database, Chinese Journal Full-text Database, and Chinese Science and Technology Periodical Database. The search language was limited to Chinese and English, and the period was from 1990 to 2018. These literatures were rigorously screened by inclusion and exclusion criteria. The data needed for this study were extracted and the Meta analysis was performed using RevMan 5.3 software. A total of 13 literatures were included, and the overall quality of the literature was relatively high, and no significant publication bias was suggested. A total of 1 994 subjects, including 720 in the SRT group and 1 274 in the CRT group. The average 3-year overall survival rates were 42.9% in SRT group and 44.8% in CRT group,with no significant difference (1.14, 95: 0.62-2.06, 0.68). The average 5-year overall survival rate (1.42, 95: 1.10-1.84, 0.01), 5-year local recurrence-free survival rate (1.68, 95: 1.11-2.55, 0.01) and 5-year local control rate (2.17, 95: 1.52-3.12, 0.01) of SRT group were 46.4%, 47.4% and 71.2%, respectively, which were higher than those of non-surgical group (37.9%, 32.0%, and 52.2% respectively). The average laryngeal function preservation rate was 19.8%,being significantly lower than 80.6% of the non-surgical group(0.03, 95: 0.01-0.07, 0.01). SRT has better long-term efficacy, while CRT has better preservation of laryngeal function.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 110-115, 2019.
Article in Chinese | WPRIM | ID: wpr-801771

ABSTRACT

Objective: To investigate transport mechanism of cyperotundone in Caco-2 cell model and provide experimental basis for clinical application of Cyperi Rhizoma. Method: The toxicity of cyperotundone with different concentrations to Caco-2 cells was investigated by methyl thiazolyl tetrazolium (MTT) colorimetry, in order to determine the concentration of administration in transport test. The content of cyperotundone was determined by liquid chromatography-mass spectrometry (LC-MS) with apparent permeability coefficient (Papp) and cumulative transport capacity as indexes. The chromatographic conditions were as following:mobile phase of acetonitrile (A)-water (B) for gradient elution (0-1.5 min, 35%A; 1.5-2 min, 35%-90%A; 2-4 min, 90%A; 4-4.1, 90%-35%A; 4.1-8 min, 35%A), the flow rate at 0.3 mL · min-1, injection volume of 1 μL, and temperature of column at 30℃. The mass spectrometric conditions was electrospray ionization (ESI) and positive ion mode, the detection ions of cyperotundone and osthole (internal standard substance) were m/z 219.2-110.9 and m/z 245.0-189.0, respectively. Effect of concentration of cyperotundone, administration time, ethylenediamine tetraacetic acid (EDTA) and P-glycoprotein (P-gp) inhibitor on the transmembrane transport of cyperotundone on in vitro cell model were investigated. Result: Cyperotundone didn't have significant toxicity to Caco-2 cells at 3-90 mg · L-1 after incubation for 4 h. The transportion of cyperotundone in Caco-2 cell model was related to the concentration and time to a certain extent, its Papp was higher than 1×10-6 cm · s-1, which indicated that absorption of cyperotundone was good, the efflux rate (ER) of cyperotundone was 0.5-1.5.There was no significant difference in bidirectional Papp of cyperotundone after the addition of cell bypass transport inhibitor (EDTA) and P-gp transport inhibitor (verapamil). Conclusion: The transport mechanism of cyperotundone in Caco-2 cell model is mainly passive diffusion, and cell bypass transport and P-gp are not involved in its transport.

3.
Journal of Xinxiang Medical College ; (12): 207-211, 2018.
Article in Chinese | WPRIM | ID: wpr-699504

ABSTRACT

Objective To investigate the effect of anesthesia at different depths on postoperative cognitive disfunction (POCD) and inflammatory response in the elderly patients undergoing abdominal operation.Methods A total of 90 elderly patients who underwent abdominal operation in the Affiliated Hospital of Shaanxi University of Chinese Medicine from June 2014 to June 2016 were divided into observation group and control group according to the depth of anesthesia,45 cases in each group.The patients in the two groups were performed with combined intravenous and inhalation anesthesia,the bispeetral index (BIS) value was maintained at 30-39 during the operation in the observation group,and the BIS value was maintained at 50-59 during the operation in the control group.The mean arterial pressure (MAP) and heart rate(HR) of patients in the two groups were recorded at the time points of entering the operation room(T0),5 minutes after tracheal cannula(T1),opening abdominal cavity (T2),closing abdominal cavity (T3) and tracheal cannula extubation (T4).The mini-mental state examination (MMSE) score of the patients in the two groups was performed before operation and the first,third,seventh day after operation;and the incidence of POCD was recorded.The levels of serum interleukin-6(IL-6) and S-100β protein were detected at the time points of before operation,the end of the operation and the first,third day after operation in the two groups.Results Five cases in the control group and six cases in the observation group were eliminated,39 cases in the observation group and 40 cases in the control group were evaluated finally.The MAP at T1 and T2 was significantly lower than that at T0 in the two groups (P < 0.05).There was no significant difference in the MAP between T3,T4 and T0 in the two groups(P < 0.05).There was no significant difference in the HR each time point in each group(P < 0.05).There was no significant difference in the MAP and HR between the two groups at each time point(P < 0.05).There was no significant difference in the MMSE score between the two groups before operation(P < 0.05).The MMSE score of patients at the first and third day after operation was significantly lower than that before operation and the seventh day after operation in the two groups (P < 0.05).There was no significant difference in the MMSE score between before operation and the seventh day after operation in the two groups(P <0.05).The MMSE score in the observation group was significantly higher than that in the control group at the first and third day after operation (P < 0.05).There was no significant difference in the MMSE score between the two groups at the seventh day after opera tion(P < 0.05).The incidences of POCD at the first,third and seventh day after operation in the observation group were 28.21% (11/39),15.38% (6/39) and 7.69% (3/39) respectively;and they were 50.00% (20/40),37.50% (15/40) and 20.00% (8/40) respectively in the control group.The incidence of POCD in the observation group was significantly lower than that in the control group at the first and third day after operation (x =3.934,4.949;P < 0.05).There was no significant difference in the incidence of POCD between the two groups at the seventh day after operation(x2 =2.496,P < 0.05).There was no significant difference in the levels of serum IL-6 and S-100β protein between the two groups before operation (P <0.05).The levels of serum IL-6 and S-100β protein at the end of operation and the first,third day after operation were significantly higher than those before operation in the two groups(P < 0.05).The levels of serum IL-6 and S-100β protein in the observation group were significantly lower than those in the control group at the end of operation and the first,third day after operation (P < 0.05).Conclusion Deep anesthesia (BIS value is maintained at 30-39) can reduce the levels of inflammatory factors,the incidence of POCD after operation and the brain damage in the elderly patients with abdominal operation.

4.
Chinese Journal of Health Policy ; (12): 76-81, 2018.
Article in Chinese | WPRIM | ID: wpr-753951

ABSTRACT

This paper reviews the development history of the Chinese Journal of Health Policy (CJHP),analyzes articles published in the journal in the past 10 years, briefly summarizes the past practices and achievements, and forecasts the future development of the journal. From October 2008 to September 2018, the journal published a total of 1 547 papers in 120 issues. This paper analyzes the distribution of the topics,the recruitment of contributions,the affiliated institutions of first authors,as well as the proportion of fund papers and the influence factors. The proportion of fund papers published in the journal increased from 41. 03% in 2008 to 82. 79% in 2018; and according to CNKI statistics, the compound influence factor in 2017 was 1. 968,the five-year influence factor was 2. 613; and the H index in 2008-2018 was 42. Over the past 10 years, the journal had made great progress, achieved good results and accumulated some experience. Its academic quality has ranked in the forefront of disciplines,has won a good academic reputation,and had been highly recognized by readers and peers. Looking forward to the future,the journal will continue to adhere to the purpose of " spreading policies,researches on policies,and providing support for policymaking",adhere to the goal of establishing high-quality professional academic journals with important influence at home and abroad,and better play the role of high-end academic platform for the dissemination and exchange of health policy research results in China.

5.
Chinese Journal of Health Policy ; (12): 72-75, 2018.
Article in Chinese | WPRIM | ID: wpr-753950

ABSTRACT

Compared with the traditional health service industry, the new health service industry tends to be more comprehensive and integrated, and the various factors are in a dynamic change process, which is characterized by larger scale,intensive,agglomerative and special business methods. Compared with the traditional medical and health industry supervision, its regulatory content has greatly changed in the new system. Under the new market relationship,the supply and demand sides have put forward higher requirements for supervision,and the emerging business forms are networked, fragmented and flexible. The service boundary has broken through the traditional supervision mode,and it simultaneously faces the challenge of defining the supervision responsibilities,complexity,the current tax system management,the current supervision mechanism hindrances,and the balance between the new health service industry and health quality access regulation which are undergoing an exponential trend of development. Therefore,this paper puts forward some suggestions such as the establishment of a new health service industry supervision mechanism with information supervision as the core,the realization of the transition from direct control to bottom-line supervision, the establishment of multi-sectoral organic coordination, and a coordinated supervision mechanism which involves the participation of all social subjects.

6.
Journal of Experimental Hematology ; (6): 1711-1715, 2014.
Article in Chinese | WPRIM | ID: wpr-340431

ABSTRACT

Compared with endothelial progenitor cells, outgrowth endothelial cells (BOECs) from peripheral blood are rich in protein for blood angiogenesis and cell adhesion, similar to mature endothelial cells in biological characteristics. Moreover, they are now replacing human umbilical vein endothelial cells for the latter's limited life span and drift of phenotype, and might become a new tool for exploring the vascular abnormalities. This study was aimed to establish the protocol of producing BOECs, and then analyze the cell phenotype and function of BOECs. Mononuclear cells were collected from peripheral blood by gradient centrifugation and then seeded on plates and cultivated in EGM-2 medium for 4 weeks. The morphological changes of cells were observed and cell phenotype was examined by flow cytometry. VWF multimers were used to analyse the distribution of vWF multimers in superment of BOECs and the storage of vWF in BOECs, and the secretion of vWF in BOECs under stimulation was detected by confocal fluorescence microscopy. The results showed that after 4-week-culture in vitro, the cell colonies and characteristic cobblestone-like morphology of BOECs were found in plates. For another three weeks of expansion, BOECs expressed CD31, CD34, and EPCR, without the expression of CD14, CD45 and CD133. The vWF from BOECs cell supernatant shared the same multimer pattern as that in normal plasma. By confocal fluorescence microscopy, vWFs were observed in BOECs. The amount of vWF increased in cells, and vWF strings were formed on cell surface by the stimulation of phorbol-12-myristate-13-acetate(PMA). It is concluded that the BOECs are first successfully established, and the phenotype and function of BOECs are analyzed. They are the native cell models for the pathogenesis of von Willebrand diseases (vWD), and may be used as new gene therapy tools for vWD.


Subject(s)
Humans , Cell Adhesion , Cell Count , Cell Culture Techniques , Methods , Cell Proliferation , Cell Separation , Methods , Cells, Cultured , Endothelial Cells , Cell Biology , Flow Cytometry , Phenotype
7.
Chinese Journal of Hematology ; (12): 751-756, 2013.
Article in Chinese | WPRIM | ID: wpr-272120

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the genetic defect and its mechanism in a patient with congenital afibrinogenemia.</p><p><b>METHODS</b>The plasma fibrinogen activity and antigen of the patient was determined using the Clauss method and immuno-nephelometric assay, respectively. Genomic DNA was isolated from peripheral blood of the proband and his related family members. All exons and exon-intron boundaries of the three fibrinogen genes (FGA, FGB, FGG) were amplified by PCR followed by direct sequencing. Thrombin fibrin aggregation curve were detected in the plasma of the patient. Wild-type and mutation type fibrinogen vectors were constructed, and then transfected into COS-7 cells. The wild-type and mutant proteins from the culture media and cell lysates were tested by Western blot and ELISA.</p><p><b>RESULTS</b>APTT, PT, TT were significantly longer in the proband. Plasma fibrinogen activity and antigen of the patient could not be detected using the Clauss method and immuno-nephelometry, respectively. Gene analysis revealed that a novel homozygous GTTT insertion between nucleotides 2833 and 2834 in FGB exon 2 in the proband. The proband's father, mother, brother and son were heterozygous. The polymerization curves of the patient did not show a lag phase or final turbidity, compared with the normal controls. Western blot analysis showed the lack of complete half-molecules of the fibrinogen molecule and fibrinogen in patient's plasma under non-reducing conditions. It also could not detect the truncated Bβ chain under reducing conditions. Abnormal fibrinogen molecule (molecule weight>340 000) were found in transfected COS-7 cells by Western blot, which indicated that the mutation caused the abnormal intracellular fibrinogen molecule assembly. The fibrinogen band was absent in culture media transfected by the mutation. Fibrinogen levels of mutant fibrinogen were no significant different from those of wild-type fibrinogen in cell lysates by ELISA analysis [(2.47 ± 0.30) μg/ml vs (2.65±0.60) μg/ml, P=0.0889]; However, the levels of the mutant fibrinogen were statistically significant lower than those of wild type fibrinogen in culture media [(0.01 ± 0.01) μg/ml vs (3.80±0.80) μg/ml, P=0.0001].</p><p><b>CONCLUSION</b>Congenital afibrinogenemia was caused by this frameshift mutation in exon 2 of FGB. This novel mutation impaired fibrinogen assembly and secretion.</p>


Subject(s)
Humans , Male , Young Adult , Afibrinogenemia , Genetics , Fibrinogen , Genetics , Frameshift Mutation , Mutagenesis, Insertional , Pedigree
8.
Chinese Journal of Gastrointestinal Surgery ; (12): 842-845, 2011.
Article in Chinese | WPRIM | ID: wpr-321225

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the diagnostic value of 3.0 T diffusion-weighted imaging with background suppression (DWIBS) magnetic resonance (MR) for lymph node metastasis in rectal cancer.</p><p><b>METHODS</b>Thirty-five patients with rectal cancer who underwent preoperative routine MRI+DWI examination were enrolled in the study and were treated by rectal cancer resection plus lymph node dissection. Metastatic and non-metastatic lymph nodes were confirmed by postoperative pathology. Apparent diffusion coefficient (ADC) values, long-axis and short-axis diameters of lymph nodes were measured. Receiver operating characteristic (ROC) curve was used to assess the diagnostic efficacy of ADC, long-axis and short-axis diameters for differentiating metastatic lymph nodes from non-metastatic lymph nodes.</p><p><b>RESULTS</b>A total of 151 lymph nodes were confirmed with exact location in 35 patients. Sixty-five metastatic lymph nodes and 86 non-metastatic lymph nodes were identified. The ADC values of metastatic lymph nodes and non-metastatic lymph nodes were(0.86±0.14)×10(-3) and (0.94±0.16)×10(-3) mm(2)/s respectively. The long-axis diameter were(9.78±3.13) and (7.90±1.77) mm, respectively. The short-axis diameter were (7.65±2.00) and (6.45±1.19) mm, respectively. There were statistically significant differences between metastatic and non-metastatic lymph nodes in ADC values, long-axis diameter, and short-axis diameter(all P<0.01). The areas under the ROC curve of ADC value, long-axis diameter, and short-axis diameter were 0.648, 0.706, and 0.692, respectively. Optimal cutoff values for these parameters were 1.05×10(-3) mm(2)/s, 7.95 mm, and 5.90 mm, respectively, and the corresponding sensitivities and specificities were 93.8% and 30.2%, 75.4% and 61.6%, 90.8% and 38.4%.</p><p><b>CONCLUSIONS</b>Quantitative measurement of ADC value may reflect the degree of diffusion restriction of metastatic lymph nodes by DWIBS at 3.0 T MR. Accurate diagnosis of metastatic lymph nodes in rectal cancer demands comprehensive evaluation combining ADC value with diameter measurement.</p>


Subject(s)
Humans , Diffusion Magnetic Resonance Imaging , Methods , Lymph Node Excision , Lymph Nodes , Pathology , Lymphatic Metastasis , Rectal Neoplasms , Diagnosis , Pathology
9.
Chinese Journal of Hematology ; (12): 154-156, 2010.
Article in Chinese | WPRIM | ID: wpr-283868

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the phenotype and genotype of a family with inherited dysfibrinogenemia.</p><p><b>METHODS</b>Assays of coagulation, including activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT), were carried out with Stago Compact in the proband and his family members. The activity and antigen of fibrinogen in plasma were determined by Clauss and immunoturbidimetry, respectively. Fibrinogen and its constituent were analyzed by Western blot with nonreducing 4%-20% SDS-polyacrylamide gel electrophoresis (PAGE). All exons and exon-intron boundaries of fibringen genes FGA, FGB and FGG were analyzed by PCR and then direct sequencing.</p><p><b>RESULTS</b>The proband had normal APTT and PT, but prolonged TT. The activity of fibrinogen in plasma was decreased while its antigen level was normal. These abnormalities were also found in his mother and a sister. Genetic analysis revealed heterozygous G1233A in the exon 2 of FGA originating from his mother, which resulted in Arg16His missense mutation.</p><p><b>CONCLUSION</b>Inherited dysfibrinogenemia was caused by Arg16His mutation in exon 2 of FGA, and this is the first case reported in a Chinese family.</p>


Subject(s)
Humans , Fibrinogen , Genetics , Genotype , Mutation , Pedigree , Phenotype
10.
Experimental & Molecular Medicine ; : 478-486, 2009.
Article in English | WPRIM | ID: wpr-107288

ABSTRACT

Osteoblasts can synthesize the insulin-like growth factors (IGFs) and the IGF-binding proteins (IGFBPs), which may either enhance or attenuate IGF-stimulated bone cell proliferation. Since estrogen induced osteoblastic differentiation and proliferation through an estrogen-responsive gene in target cells, we investigated the effects of estrogen on IGFBP-6 expression in the human osteoblastic-like cell line SaOS-2. Expressions of IGFBP-6 protein and mRNA increased 2.8 and 2-fold, respectively, in the presence of 17-beta-estradiol (E2) (0.01 to 1 micrometer) and estrogen receptor (ER) in SaOS-2 cells. On the other hand, E2 induced a 2-fold increase in SaOS-2 cell proliferation. To identify genomic sequences associated with estrogen responsiveness, the 5'-promoter region (-44 to +118) of the IGFBP-6 gene was cloned into a chloramphenicol acetyltransferase (CAT) reporter vector. E2 induced a 3-fold increase in CAT activity in SaOS-2 cells transiently transfected with this construct. Identification of the estrogen-responsive element (ERE) [5'-CCTTCA CCTG-3'] (-9 to +1) in this IGFBP-6 gene promoter region was confirmed using electromobility shift assays and deletion analysis. This functional ERE was important for E2-induced trans-activation of the IGFBP-6 gene. These results demonstrate that E2 exhibits a positive effect on IGFBP-6 gene transcription through estrogen-liganded ER binding to the functional ERE in the IGFBP-6 gene promoter in SaOS-2 cells.


Subject(s)
Humans , Blotting, Western , Cell Proliferation , Chloramphenicol O-Acetyltransferase/metabolism , Electrophoretic Mobility Shift Assay , Estradiol/pharmacology , Estrogen Receptor alpha/genetics , Estrogens/pharmacology , Insulin-Like Growth Factor Binding Protein 6/genetics , Osteoblasts/drug effects , Promoter Regions, Genetic/genetics , RNA, Messenger/genetics , Response Elements , Reverse Transcriptase Polymerase Chain Reaction , Transcriptional Activation , Tumor Cells, Cultured
11.
Chinese Journal of Hematology ; (12): 145-148, 2008.
Article in Chinese | WPRIM | ID: wpr-262918

ABSTRACT

<p><b>OBJECTIVE</b>To identify gene mutations involved in five cases of inherited factor V (FV) deficiency.</p><p><b>METHODS</b>Activity of FV was determined by one-stage clotting assay using FV-deficiency plasma, and FV antigen by an ELISA assay. All the exons and exon-intron boundaries of FV gene were amplified by PCR and then DNA sequencing. Restriction enzyme analysis was used to analyze the probands, their family members and healthy volunteers.</p><p><b>RESULTS</b>Both activity and antigen of FV in the 5 patients were extremely lower compared with that of normal mixed plasma. Six mutations were identified in these 5 patients, G69969T (G2079V), C45533T (R712Ter), C46796T (R1133Ter), G45366A (C656Y), C46253T (R952C) and G16088C (D68H), the latter three were novel mutations reported for the first time and the C46253T (R952C) was the first missense mutation reported in B domain. The result of sequencing or restriction enzyme analysis showed that the three novel missense mutations were not caused by single nucleotide polymorphisms.</p><p><b>CONCLUSION</b>Gene mutations in 5 type I inherited FV deficiency of patients including 2 nonsense mutations and 4 missense mutations identified which led to the instability of FV protein and the reducing of FV: Ag in the plasma.</p>


Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Young Adult , DNA Mutational Analysis , Exons , Genetics , Factor V , Genetics , Metabolism , Factor V Deficiency , Blood , Genetics , Mutation , Pedigree , Phenotype
12.
Journal of Experimental Hematology ; (6): 871-874, 2008.
Article in Chinese | WPRIM | ID: wpr-267869

ABSTRACT

The study was purposed to investigate the expression and function of non-muscle myosin heavy chain-IIA (NMMHC-IIA) in Fechtner syndrome in order to explore the pathologic changes of kindy disease and the mechanism of granulocyte inclusion body formation. NMMHC-IIA levels in granulocytes were analyzed by Western-blot, the expressions of NMMHC-IIA, IIB in HEK-293 cells were detected by RT-PCR and were analyzed by co-immunoprecipitation. The results indicated that the IIA/beta-actin ratio for Fechtner syndrome granulocytes was (0.35 +/- 0.12), and obviously decreased as compared with that of normal control (0.87 +/- 0.18) (p < 0.01). The IIA and IIB expressed higher in HEK-293 cells. The interaction of IIA and IIB was confirmed by co-immunoprecipitation in HEK-293 cells. It is concluded that dominant-negative effect of NMMHC-IIA is involved in the formation of inclusion bodies. IIA and IIB show obvious interaction, IIB partly compensates the IIA defect derived from MYH9 mutations, and may delay or prevent the development of clinically relevant abnormalities.


Subject(s)
Humans , Blood Platelet Disorders , Genetics , Metabolism , Pathology , Cell Line , Granulocytes , Pathology , Inclusion Bodies , Pathology , Kidney , Cell Biology , Embryology , Metabolism , Mutation , Nonmuscle Myosin Type IIA , Genetics , Metabolism , Physiology , Nonmuscle Myosin Type IIB , Genetics , Metabolism , Physiology , Syndrome , Thrombocytopenia , Genetics , Metabolism , Pathology
13.
Chinese Journal of Hematology ; (12): 152-155, 2007.
Article in Chinese | WPRIM | ID: wpr-328394

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical manifestations, pathologic features and laboratory findings in two Proteus syndrome patients with giant hemangiomas in the spleen and chronic DIC.</p><p><b>METHODS</b>Ultrasound imaging and magnetic resonance imaging (MRI) were used for analysing the characteristics of the giant hemangiomas in the spleen. The spleen specimen was examined pathologically for the feature of the hemangioma. Homostatic tests were performed by routine laboratory methods.</p><p><b>RESULTS</b>Two Proteus syndrome patients with giant hemangiomas in the spleen causing chronic DIC (Kasabach-Merritt syndrome) were first reported. They were recovered after splenectomy.</p><p><b>CONCLUSION</b>Proteus syndrome when accompanied giant hemangioma could cause chronic DIC. Significantly decreased plasma fibrinogen level in this case might be helpful for the differential diagnosis from DIC caused by other diseases.</p>


Subject(s)
Adolescent , Female , Humans , Disseminated Intravascular Coagulation , Hemangioma, Cavernous , Diagnostic Imaging , General Surgery , Proteus Syndrome , Splenectomy , Splenic Neoplasms , Diagnostic Imaging , General Surgery , Ultrasonography
14.
Journal of Experimental Hematology ; (6): 416-419, 2004.
Article in Chinese | WPRIM | ID: wpr-352051

ABSTRACT

To establish leukemic cell lines stably transfected by RbAp46 gene, electroporation was performed after optimizing the transfection condition for suspended cells. Under conditions of low voltage and high capacitance, RbAp46 was transfected into U937 by electroporation. Individual clones selected with G418 for 3 weeks were isolated. The integration and the protein levels of the exogenous RbAp46 in transfectants were determined by PCR and Western blot analysis, respectively. The subclone expressing high level of RbAp46 was then established. Viability of transfected cells was assayed by trypan blue exclusion. Cell number was counted daily to determine the growth rate. The results showed that growth rate of U937 cell lines expressing exogenous RbAp46 was about 50% lower than that in control. It is concluded that leukemic cell lines stably expressing exogenous RbAp46 were established and overexpression of RbAp46 inhibits the growth of U937 leukemic cells.


Subject(s)
Humans , Blotting, Western , Carrier Proteins , Genetics , Cell Proliferation , Electroporation , Nuclear Proteins , Genetics , Retinoblastoma-Binding Protein 7 , Transfection , U937 Cells , WT1 Proteins , Genetics
15.
Chinese Journal of Hematology ; (12): 480-483, 2003.
Article in Chinese | WPRIM | ID: wpr-354847

ABSTRACT

<p><b>OBJECTIVE</b>To identify a mutation G2113-->A in the glycoprotein (GP)IX gene associated with Bernard-Soulier syndrome (BSS) and to investigate BSS pathogenesis.</p><p><b>METHODS</b>Allele-specific restriction enzyme was used to analyze the samples of patient, her mother, her brother and 40 healthy volunteers. Site-directed mutagenesis was performed to construct a expression vector PD-IXG2113A harboring the mutation G2113-->A. Chinese hamster ovary (CHO) cells were transiently cotransfected with plasmids harboring the entire coding region of GPIbalpha, GPIbeta and GPIX or mutant GPIX, respectively. Expression of GPIbalpha and GPIX in transfected CHO cells were analysed with flow cytometer. GPIbalpha and GPIX in the cytoplasma of transfected CHO cells were analysed by immunostaining and Western blotting.</p><p><b>RESULTS</b>The patient was found to be homozygosity of the substitution, her mother and her brother be heterozygous. Expressions of GPIbalpha and GPIX in mutant CHO cells were remarkably reduced, but abundant in the cytoplasma.</p><p><b>CONCLUSION</b>The mutation of Ala139(GCC)-->Thr(ACC) in the GPIX did not affect synthesis and assembly of GPIb/IX complex but influence its anchoring and expression on the cell surface, which was responsible for BSS.</p>


Subject(s)
Adult , Animals , Cricetinae , Female , Humans , Bernard-Soulier Syndrome , Genetics , Blotting, Western , CHO Cells , Mutation , Platelet Glycoprotein GPIb-IX Complex , Genetics
SELECTION OF CITATIONS
SEARCH DETAIL