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1.
Journal of Experimental Hematology ; (6): 192-196, 2018.
Article in Chinese | WPRIM | ID: wpr-278697

ABSTRACT

<p><b>OBJECTIVE</b>To explore the expression and clinical significance of VEGF, IL-17, β2-MG and IL-35 in patients with multiple myeloma.</p><p><b>METHODS</b>A total of 83 patients with multiple myeloma (MM) from January 2012 to December 2016 were enrolled in MM group, 36 healthy subjects were enrolled in control group. The levels of IL-17, IL-35 and VEGF in serum were detected by ELISA. The levels of β2-MG in serum were measured by radioimmunoassay. The differences of different indexes between 2 groups were compared.</p><p><b>RESULTS</b>The serum levels of IL-17, VEGF and β2-MG in serum of III stage were higher than that in II stage, which was higher than that in I stage and control group (P<0.05). The levels of IL-35 in the control group were significantly higher than those in the I,II,III stage group (P<0.05). The levels of IL-17, VEGF and β2-MG in serum of progress period were higher than those in stable phase and control group, level of IL-35 in serum of control group was significantly higher than that in the stable phase and progress period group (P<0.05). The correlation analysis showed that the level of serum IL-17 positively correlated with VEGF, β2-MG expression (r=0.65, 0.58, P<0.05); and the serum IL-17 levels were negatively correlated with IL-35 levels (r=-0.42, P<0.05).</p><p><b>CONCLUSION</b>The anomalous levels of IL-17, IL-35, IVEGF and β2-MG expressions correlate with the progression and prognosis of patients with multiple myeloma.</p>

2.
Journal of Kunming Medical University ; (12): 117-119, 2018.
Article in Chinese | WPRIM | ID: wpr-694544

ABSTRACT

Objective To carry out the investigation of common susceptibility factors of primary hypertension in young people,and to find the pathogenic factors of primary hypertension. Methods 354 cases of young patients with essential hypertension were selected from 7360 young students in School of CAPF Frontier Guards Force and compared to 115 young healthy students at the same age. Results The incidence of primary hypertension is increasing year by year. More common risk factors are seen among students with hypertension compared to those of the control group. Conclusion Risk factors of primary hypertension are commonly seen among young students in School of CAPF Frontier Guards Force. It is necessary to prevent the risk factors, apply the targeted intervention and improve the management and issue a plan for prevention and control of high blood pressure.

3.
Chinese Medical Equipment Journal ; (6): 94-98, 2018.
Article in Chinese | WPRIM | ID: wpr-700002

ABSTRACT

The concept of biometric identification technology (BIT)was described,and several typical BITs were introduced such as identification technologies for hand, face, physiological features and behavioral trait. The application of BIT was analyzed in military field of foreign countries and China.It's pointed out BIT's future involved in bioassay,integration with wireless video identification technique and etc,comprehensive identification technique with high throughput,high precision and multi mechanism as well as enhanced safety.

4.
Chinese Journal of Contemporary Pediatrics ; (12): 1169-1173, 2017.
Article in Chinese | WPRIM | ID: wpr-300427

ABSTRACT

The clinical manifestations of five children with paroxysmal kinesigenic dyskinesia (PKD) were retrospectively analyzed and their gene mutations were analyzed by high-throughput sequencing and chromosome microarray. The 5 patients consisted of 4 males and 1 female and the age of onset was 6-9 years. Dyskinesia was induced by sudden turn movement, scare, mental stress, or other factors. These patients were conscious and had abnormal posture of unilateral or bilateral extremities, athetosis, facial muscle twitching, and abnormal body posture. The frequency of onset ranged from 3-5 times a month to 2-7 times a day, with a duration of <30 seconds every time. Electroencephalography showed no abnormality in these patients. Three patients had a family history of similar disease. The high-throughput sequencing results showed that a heterozygous mutation in the PRRT2 gene, c.649_650insC (p.R217PfsX8), was found in two patients; the mutation c.436C>T (p.P146S) was found in one patient; a splice site mutation, IVS2-1G>A, was found in one patient. The two mutations c.436C>T and IVS2-1G>A had not been reported previously. The chromosome microarray analysis was performed in one patient with negative results of gene detection, and the chromosome 16p11.2 deletion (0.55 Mb) was observed. Low-dose carbamazepine was effective for treatment of the 5 patients. PKD is a rare neurological disease. The detection of the PRRT2 gene by multiple genetic analysis can help the early diagnosis of PKD.


Subject(s)
Child , Female , Humans , Male , Carbamazepine , Therapeutic Uses , Chromosome Deletion , Chromosomes, Human, Pair 16 , Dystonia , Diagnosis , Drug Therapy , Genetics , Electroencephalography , Membrane Proteins , Genetics , Mutation , Nerve Tissue Proteins , Genetics
5.
Chinese Medical Equipment Journal ; (6): 118-121, 2017.
Article in Chinese | WPRIM | ID: wpr-699875

ABSTRACT

Objective To discuss the importance of internal environment and facilities in the clinical laboratory.Methods The design and construction requirements of internal environment and facilities were introduced from the aspects of laboratory equipment selection,installation site and amount as well as construction materials and etc.Results Internal environment and facilities were very important for clinical laboratory,which determined its applicability,comfort,quality and efficiency of the clinical laboratory.Conclusion Design and construction of environment and facilities for clinical laboratory must be scientific and reasonable.

6.
Chinese Medical Journal ; (24): 941-947, 2015.
Article in English | WPRIM | ID: wpr-350374

ABSTRACT

<p><b>BACKGROUND</b>MicroRNAs (miRNAs) function as essential posttranscriptional modulators of gene expression, and are involved in a wide range of physiologic and pathologic states, including cancer. Numerous miRNAs are deregulated in hepatocellular carcinoma (HCC). This study aimed to investigate the role of miR-27a in the development of HCC.</p><p><b>METHODS</b>The expression of MiR-27a was measured by quantitative real-time polymerase chain reaction (qRT-PCR). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to examine changes in the viability of HepG2, Bel-7402, Bel-7404 hepatoma cell lines associated with up-regulation or down-regulation of miR-27a. A dual-luciferase activity assay was used to verify a target gene of miR-27a. Immunohistochemistry, qRT-PCR, Western blotting analysis, and cell cycle and apoptosis flow cytometric assays were used to elucidate the mechanism by which miR-27a modulates liver cancer cell proliferation.</p><p><b>RESULTS</b>The expression of miR-27a was significantly increased in HCC tissues and HepG2, Bel-7402, Bel-7404 hepatoma cell lines (P < 0.05). We also found that the down-regulation of miR-27a in HepG2 cells dramatically inhibited proliferation, blocked the G1 to S cell cycle transition and induced apoptosis (P < 0.05). In addition, miR-27a directly targeted the 3'- untranslated region of peroxisome proliferator-activated receptor γ (PPAR-γ), and ectopic miR-27a expression suppressed PPAR-γ expression on the mRNA and protein levels. The rosiglitazone-induced overexpression of PPAR-γ attenuated the effect of miR-27a in HCC cells.</p><p><b>CONCLUSIONS</b>Our findings suggested that miRNA-27a promoted HCC cell proliferation by regulating PPAR-γ expression. MiR-27a may provide a potential therapeutic strategy for HCC treatment.</p>


Subject(s)
Humans , Carcinoma, Hepatocellular , Genetics , Metabolism , Cell Proliferation , Genetics , Physiology , Gene Expression Regulation, Neoplastic , Hep G2 Cells , Liver Neoplasms , Genetics , Metabolism , MicroRNAs , Genetics , Physiology , PPAR gamma , Metabolism
7.
Braz. j. microbiol ; 45(1): 351-358, 2014. ilus
Article in English | LILACS | ID: lil-709455

ABSTRACT

Laccases are blue copper oxidases (E.C. 1.10.3.2) that catalyze the one-electron oxidation of phenolics, aromatic amines, and other electron-rich substrates with the concomitant reduction of O2 to H2O. A novel laccase gene pclac2 and its corresponding full-length cDNA were cloned and characterized from Phytophthora capsici for the first time. The 1683 bp full-length cDNA of pclac2 encoded a mature laccase protein containing 560 amino acids preceded by a signal peptide of 23 amino acids. The deduced protein sequence of PCLAC2 showed high similarity with other known fungal laccases and contained four copper-binding conserved domains of typical laccase protein. In order to achieve a high level secretion and full activity expression of PCLAC2, expression vector pPIC9K with the Pichia pastoris expression system was used. The recombinant PCLAC2 protein was purified and showed on SDS-PAGE as a single band with an apparent molecular weight ca. 68 kDa. The high activity of purified PCLAC2, 84 U/mL, at the seventh day induced with methanol, was observed with 2,2'-azino-di-(3-ethylbenzothialozin-6-sulfonic acid) (ABTS) as substrate. The optimum pH and temperature for ABTS were 4.0 and 30 ºC, respectively . The reported data add a new piece to the knowledge about P. Capsici laccase multigene family and shed light on potential function about biotechnological and industrial applications of the individual laccase isoforms in oomycetes.


Subject(s)
Laccase/genetics , Laccase/metabolism , Phytophthora/enzymology , Cloning, Molecular , Conserved Sequence , Enzyme Stability , Gene Expression , Hydrogen-Ion Concentration , Laccase/chemistry , Laccase/isolation & purification , Molecular Weight , Open Reading Frames , Protein Structure, Tertiary , Phytophthora/genetics , Pichia/genetics , Protein Sorting Signals/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Sequence Homology, Amino Acid , Temperature
8.
Chinese Medical Journal ; (24): 4348-4353, 2013.
Article in English | WPRIM | ID: wpr-327573

ABSTRACT

<p><b>OBJECTIVE</b>We aimed to examine the current developments and challenges confronted by old age security in rural China.</p><p><b>DATA SOURCES</b>This study is based on the data from PubMed, Elsevier, Wiley, EBSCO, EMBASE, SCI Expanded, ProQuest, Google, and CNKI which is the most informative database in Chinese.</p><p><b>STUDY SELECTION</b>Articles were selected with the search terms "rural", "China", "old", "older", or "elder", "elderly", or "aged", "aging", "security", "culture", "value", "medical insurance" or "community based medical insurance" or "cooperative medical scheme". Related websites and yearbooks were searched as well.</p><p><b>RESULTS</b>The socio-economic development has made the burden of traditional care for the rural elderly heavier than ever, and new challenges are emerging in rural communities, such as poor economic, deteriorating natural environment and health crisis.</p><p><b>CONCLUSIONS</b>The governments should improve the scale and caliber of rural old age security and strengthen regulations with great efforts in developing the rural economy and protecting the natural environment of rural communities.</p>


Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , China , Old Age Assistance , Rural Population
9.
Chinese Medical Journal ; (24): 2663-2670, 2012.
Article in English | WPRIM | ID: wpr-244375

ABSTRACT

<p><b>BACKGROUND</b>Chromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.</p><p><b>METHODS</b>All 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.</p><p><b>RESULTS</b>The analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.</p><p><b>CONCLUSIONS</b>Chinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , China , Chromosome Aberrations , Chromosomes, Human, Pair 1 , Genetics , Cytogenetic Analysis , In Situ Hybridization, Fluorescence , Karyotyping , Multiple Myeloma , Genetics , Pathology
10.
Chinese Journal of Contemporary Pediatrics ; (12): 377-381, 2010.
Article in Chinese | WPRIM | ID: wpr-347594

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the dynamic changes of lipoprotein lipase (LPL) expression in the hippocampus of epileptic rats and to study its effect on vitamin E levels in rats following status epilepticus (SE).</p><p><b>METHODS</b>Rat model of SE was induced by intraperitoneal injection of pilocarpine. The rats receiving an injection of normal saline were used as a control group. The expression of LPL in the hippocampal tissue was determined using immunofluorescent methods and the level of vitamin E was examined by the colormeric method 12 hrs, 24 hrs, 3 days, 7 days and 14 days after SE.</p><p><b>RESULTS</b>LPL was expressed in the control and SE groups. In the SE group, the LPL expression began to increase 24 hr after SE (P<0.05), reached a peak 3 days after SE (P<0.01), and kept at a high level 7 days after SE (P<0.01). By 14 days, the LPL expression was reduced to the level similar to the control group. The level of vitamin E began to decline 12 hrs after SE (P<0.01), and decreased to a nadir 24 hrs after SE (P<0.01). At 3 and 7 days after SE, the levels of vitamin E were still significantly lower than the controls (P<0.05). By 14 days, the vitamin E level increased to the level similar to the control group.</p><p><b>CONCLUSIONS</b>The over-expression of LPL in the hippocampus may play an important role in the oxidative stress mechanisms following SE by regulating the uptake of vitamin E.</p>


Subject(s)
Animals , Male , Rats , Fluorescent Antibody Technique , Hippocampus , Metabolism , Lipoprotein Lipase , Physiology , Oxidative Stress , Rats, Sprague-Dawley , Status Epilepticus , Metabolism , Vitamin E , Metabolism
11.
Electron. j. biotechnol ; 12(2): 9-10, Apr. 2009. ilus, tab
Article in English | LILACS | ID: lil-551370

ABSTRACT

This paper describes a simple, low cost and reliable DNA template preparation protocol for polymerase chain reaction (PCR) using immature leaves from peanut seeds or leaves from field-grown plants. The technique may find wide utility in studies involving PCR-based molecular markers, rapid screening for transformants and gene cloning.


Subject(s)
Arachis/enzymology , Arachis/genetics , Arachis/chemistry , Polymerase Chain Reaction/economics , Polymerase Chain Reaction , DNA, Plant/analysis , DNA, Plant/chemical synthesis , Genetic Markers , Guidelines as Topic/analysis
12.
Chinese Journal of Contemporary Pediatrics ; (12): 288-290, 2009.
Article in Chinese | WPRIM | ID: wpr-347935

ABSTRACT

<p><b>OBJECTIVE</b>To study serum levels of melatonin in children with epilepsy or febrile seizures in order to provide a basis for the treatment of epilepsy or febrile seizures with melatonin.</p><p><b>METHODS</b>Serum melatonin levels were measured using ELISA in 15 children with simple febrile seizure (SFS), in 15 children with complex febrile seizure (CFS), in 15 children with epilepsy, and in 15 children with upper respiratory infections (control group).</p><p><b>RESULTS</b>Serum melatonin levels in children with epilepsy (8.66+/-1.38 ng/L) or CFS (14.91+/-2.61 ng/L) were significant lower than those in the control group (23.93+/-2.01 ng/L) (P<0.01). The SFS group showed lower serum melatonin levels (20.72+/-2.54 ng/L) compared with the control group, but there were no statistical differences between the two groups. Serum melatonin levels in the epilepsy group were significantly lower than those in the CFS (P<0.05) and the SFS groups (P<0.01).</p><p><b>CONCLUSIONS</b>Serum melatonin levels decreased in children with epilepsy or CFS. Supplement of exogenous melatonin might be a promising treatment for epilepsy and febrile seizures in children.</p>


Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Epilepsy , Blood , Drug Therapy , Melatonin , Blood , Pharmacology , Therapeutic Uses , Seizures, Febrile , Blood
13.
Chinese Acupuncture & Moxibustion ; (12): 498-500, 2009.
Article in Chinese | WPRIM | ID: wpr-257951

ABSTRACT

Two kinds of divided methods currently used in clinic and their unified new method on the scalp acupuncture were discussed. Through comparing with the nomination and selecting methods on the stimulating location, the characteristics of the two kinds of divided methods on the location of scalp acupuncture were pointed out respectively. The advantage of the method with scalp projection areas is that the nomination is easy to associate and remember, however its disadvantage is that the measurement on point-selecting is less reasonable. For the method with standard treatment lines, its measurement is more reasonable, but its nomination on the location is sophisticated and difficult to be associated and remembered, especially having more repeated lines. The new divided method of scalp acupuncture combines the advantages of the two methods in order to easily remember and freely apply through their unification, which is worth being popularized.


Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , Methods , Reference Standards , Randomized Controlled Trials as Topic , Methods , Reference Standards , Scalp
14.
Chinese Journal of Preventive Medicine ; (12): 601-606, 2009.
Article in Chinese | WPRIM | ID: wpr-316128

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of methylation status of CpG islands of endogenous E-cadherin (CDH1) gene on the promoter activity of corresponding genes in reporter assays.</p><p><b>METHODS</b>The methylation statuses of CpG island of CDH1 in 8 different cell lines were detected by methylation-specific PCR. CDH1 protein was analyzed by Western blotting. Two sets of pGL3 reporter vectors with different genotypes/haplotypes of the CDH1 promoter were constructed [ pGL3-A(-73)/-C(-73) pGL3-H1/-H4] and used to transfect these cell lines. The differences between these promoter reporter vectors were analyzed by t-test.</p><p><b>RESULTS</b>(1) CDH1 CpG island was unmethylated in AGS, MCF7, MKN74, and PC-3 cell lines, expressed in MCF7, MKN74, and PC-3, but not in AGS. Expression of CDH1 was silenced by methylation in HeLa, BGC823, A549, and RKO cell lines. (2) In the four CDH1-unmethylated MCF7, MKN74, PC-3, and AGS cell lines, the promoter activities of pGL3-C(-73), (as 0.78 +/- 0.10, 0.17 +/- 0.01, 0.11 +/- 0.01, 1.19 +/- 0.18) were significantly higher than those of pGL3-A(-73) (as 0.30 +/- 0.08, 0.07 +/- 0.01, 0.07 +/- 0.01, 0.39 +/- 0.04) (t values are -6.298, - 12.349, -8.128, -7.388, and P <. 0.1). However, in the four CDH1-methylated HeLa, BGC823, A549, and RKO cell lines, the promoter activity of pGL3-C(-73) (as 0.09 +/- 0.02, 0.13 +/- 0.02, 0.05 +/- 0.01, 0.01 +/- 0.00) was significantly lower than that of pGL3-A(-73) (as 0.16 +/- 0.01, 0.25 +/- 0.01, 0.11 +/- 0.03, 0.03 +/- 0.00) (t valued at 5.958, 11.189, 3.661, 13.866, and P<0.05). (3) In the unmethylated MKN74 and methylated RKO cell lines, the promoter activities of pGL3-H1/-H4 were obviously and contrarily different (as 1.57 +/- 0.23/0.94 +/- 0.06 and 0.38 +/- 0.02/0.50 +/- 0.04, t values were 4.577 and -4.915, P values were 0.010 and 0.003).</p><p><b>CONCLUSION</b>The methylation status of CpG island of the target gene in the tested cell lines affects the promoter activity in Reporter Assay significantly. The most active one may be the most suppressive one.</p>


Subject(s)
Humans , Cadherins , Genetics , Metabolism , Cell Line, Tumor , CpG Islands , Genetics , DNA Methylation , Flow Cytometry , Genes, Reporter , Luciferases , Genetics , Methyl-CpG-Binding Protein 2 , Genetics , Promoter Regions, Genetic
15.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 130-135, 2009.
Article in Chinese | WPRIM | ID: wpr-245943

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of nasal mucosa fibrosis on radiation induced nasal mucosa injury.</p><p><b>METHODS</b>Seventy two male rats were randomly divided into two groups, control group and irradiation injured group (radiation dose were 40 Gy); the rats were killed 1 week, 2 weeks, 4 weeks, 8 weeks, 3 months and 6 months after the finish of radiation. The middle turbinates of the animals were removed. The pathological change of the nasal mucosa were observed with scanning electron microscope, transmission electron microscope, hematoxylin and eosin (HE), alcian blue-periodic acid-Schif (AB-PAS), and Masson Trichrome (MT). The Hyp content in nasal mucosa was measured with chemo-chromatometry.</p><p><b>RESULTS</b>After radiation, the pathological characteristics in early stage (within 4 weeks) was acute inflammatory reaction. The repair of nasal mucosa started 4 weeks after radiation, lasted to 6 months. The deposition of collagen in nasal mucosa could be found 1 week after irradiation and increased gradually.</p><p><b>CONCLUSION</b>Irradiation could induce a serials of pathological changes on nasal mucosa. The nasal mucosa fibrosis may be one of the reasons of persistent irradiation induced nasal mucosa injury.</p>


Subject(s)
Animals , Male , Rats , Fibrosis , Nasal Mucosa , Pathology , Radiation Injuries, Experimental , Pathology , Rats, Sprague-Dawley , Wound Healing
16.
Journal of Experimental Hematology ; (6): 1182-1185, 2007.
Article in Chinese | WPRIM | ID: wpr-318762

ABSTRACT

This study was aimed to investigate the effect of STI571, an inhibitor of tyrosine kinase, on the proliferation and apoptosis of chronic myelogenous leukemia (CML) cells as well as expression of anti-apoptotic protein Mcl-1, and to explore the possible role of Mcl-1 in apoptosis-inducing mechanism. K562 cell line was used to observe the effect of STI571 on CML cells. Proliferation and cytotoxicity were analyzed by MTT assay. The apoptotic cells were labelled with Annexin V-FITC and PI and then analyzed by flow cytometry. The expression of apoptotic-related proteins in K562 cells was determined by Western blot with specific antibodies. The results showed that STI571 significantly inhibited the proliferation and induced apoptosis of K562 cells in a dose-and time-dependent manner. Coincidently, the protein phosphorylation on tyrosine residues was reduced and the expressions of anti-apoptotic protein Mcl-1 and Bcl-xl were down-regulated after exposure to STI571. It is concluded that STI571 induces the apoptosis of CML cells by down-regulating the expressions of Mcl-1 and Bcl-xl, which suggests that Mcl-1 and Bcl-xl may play an important role in anti-apoptotic process of CML cells.


Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , Benzamides , Down-Regulation , Gene Expression Regulation, Neoplastic , Imatinib Mesylate , K562 Cells , Myeloid Cell Leukemia Sequence 1 Protein , Phosphorylation , Piperazines , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Pyrimidines , Pharmacology , bcl-X Protein , Metabolism
17.
Chinese Journal of Preventive Medicine ; (12): 43-46, 2007.
Article in Chinese | WPRIM | ID: wpr-290197

ABSTRACT

<p><b>OBJECTIVE</b>To compare binding activity of different zinc finger domain of human Kaiso with methylated CpG.</p><p><b>METHODS</b>pGEX constructs with different human Kaiso domain were generated and then corresponding fusion proteins were induced and purified. Electrophoretic mobility shift assays were applied to evaluate the binding activity of fusion proteins with methylated CpG.</p><p><b>RESULTS</b>The purified GST-KaisoZF fusion protein (without the POZ protein binding domain) could bind with methylated CpG probe specifically, but not for three or two zinc fingers without flanking domains.</p><p><b>CONCLUSION</b>Human zinc finger protein Kaiso could bind with methylated CpG specifically, only in the assistance of the neighboring flank sequence of the zinc finger domain.</p>


Subject(s)
Humans , Base Sequence , CpG Islands , DNA Methylation , Transcription Factors , Genetics , Zinc Fingers , Genetics
18.
Chinese Journal of Biotechnology ; (12): 915-918, 2007.
Article in Chinese | WPRIM | ID: wpr-276187

ABSTRACT

Adenovirus vectors are one of the most promising gene transfer systems. They are of great value for gene therapy because these vectors achieve temporal high-level transgene expression and high gene transfer efficiency. To meet increasing needs of adenovirus vectors for gene therapy programs, parallel development of efficient, scalable and reproducible production processes is required. Perfusion cultivation of 293 cells is one of the most commonly used methods to produce adenovirus vectors and it is suitable for industrialized production specially. Experimental studies had been carried out to produce recombinant adenovirus containing the green fluorescent protein gene (Ad-GFP) by perfusion cultivation of HEK-293 N3S cells in a 5L stirring bioreactors. Perfusion rate was 1-2 volume/day. To infect the 293 N3S cells with Ad-GFP at the density of (2-4) x 10(6) cells/ ml. The time of collecting cells was 48 hours post infection. After three rounds of freeze/thaw and centrifugation, the crude viral lysates were stored at--80 degrees C until use. Then to get the Ad-GFP products by 2 x CsCl-gradient purification. The purity of the products was determined by the A260/A280 ratio and a high performance liquid chromatography (HPLC) assay. The infective titer was determined by a TCID50 assay. The culture term was 10-12 days. The infectious titer, the number of virus particle and the ratio of infectious titer to virus particle for the product were 1.0 x 10(11) IU/mL, 1.68 x 10(12) VP/mL and 6.0% IU/VP respectively. The A260/A280 ratio was 1.33, and the purity determined by HPLC was 99.2%. The cell specific productivity was around 1000 IU/cell. By perfusion cultivation of 293 N3S cells in a 5L stirring bioreactors, we established the production process for Ad-GFP, which paves a way to produce other recombinant adenovirus for gene therapy.


Subject(s)
Humans , Adenoviridae , Genetics , Bioreactors , Microbiology , Cell Line , Gene Transfer Techniques , Genetic Vectors , Green Fluorescent Proteins , Genetics , Kidney , Cell Biology , Virology , Recombinant Proteins , Genetics , Recombination, Genetic , Virus Cultivation , Methods
19.
Journal of Zhejiang University. Science. B ; (12): 346-352, 2005.
Article in English | WPRIM | ID: wpr-249209

ABSTRACT

The performance of the Dongying multi-stage ponds-wetlands ecosystem was investigated in this work. Study of the removal of different pollutants (BOD(5), COD, SS, TP, TN, NH(3)-N, etc.) in different temperature seasons and different units in this system indicated that effluent BOD(5) and SS were constant to less than 11 mg/L and 14 mg/L throughout the experimental processes; but that the removal efficiencies of pollutants such as TP, TN, NH(3)-N, COD varied greatly with season. The higher the temperature was, the higher was the observed removal in this system. Additionally, each unit of the system functioned differently in removing pollutants. BOD(5) and SS were mainly removed in the first three units (hybrid facultative ponds, aeration ponds and aerated fish ponds), whereas nitrogen and phosphates were mainly removed in hydrophyte ponds and constructed reed wetlands. The multi-stage ponds-wetlands ecosystem exhibits good potential of removing different pollutants, and the effluent quality meet several standards for wastewater reuse.


Subject(s)
Ecosystem , Fresh Water , Chemistry , Nitrogen , Phosphates , Temperature , Waste Disposal, Fluid , Water , Chemistry , Water Purification , Methods
20.
Academic Journal of Second Military Medical University ; (12): 306-307, 2001.
Article in Chinese | WPRIM | ID: wpr-736835

ABSTRACT

Objective: To understand the effect of dobutamine on the integrated backscatter of myocardium. Methods: Nineteen open-chest dogs were detected by HP Sonos 5500 with S8 transducer, and the WT, AIBS and CVIB were measured before and after the dobutamine stress. Results: During the dobutamine stress, following the increase of WT, the transmural, subepicardial and subendocardial CVIB all increased. And the increase of subepicardial CVIB was greater than that of subendocardial CVIB, leading to the decrease of TGI. But the AIBS didn't change significantly in the whole process. Conclusion: Dobutamine can enhance myocardium CVIB but has no obvious infleunce on AIBS.

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