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1.
International Eye Science ; (12): 2155-2158, 2020.
Article in Chinese | WPRIM | ID: wpr-829724

ABSTRACT

@#AIM: To evaluate the changes of anterior chamber structure in patients after combined surgery of glaucoma and cataract with viscoelastics injection under the sclera valve by OCT. <p>METHODS: The cataract with acute glaucoma patients who underwent combined surgery of glaucoma and cataract from 2016 to 2018 were enrolled. The patients were divided into the viscoelastics group and the control group. The anterior chamber depth(ACD), the angle opening distance(AOD<sub>500</sub>)and the trabecular iris angle(TIA)were examined to compare the changes of the anterior chamber structure between the two groups. <p>RESULTS: At 1 and 3d after surgery, the ACD(3.82±0.51,3.71±0.63, 3.78±0.33mm), the angle opening distance(AOD<sub>500</sub>: 0.25±0.04, 0.24±0.04, 0.25±0.05mm)and the TIA(25.13°±8.06°, 26.18°±8.06°, 29.25°±6.22°)were statistically different between two groups. There was no significant difference on the ACD, the AOD<sub>500</sub> and the TIA of the two groups 7d after surgery(<i>P</i><0.05).<p>CONCLUSION: Viscoelastics injection under the sclera valve can deepen the anterior chamber effectively after combined surgery of glaucoma and cataract. It avoids the occurrence of shallow anterior chamber in the early postoperation and maintains the opening of anterior chamber.

2.
China Journal of Chinese Materia Medica ; (24): 1471-1475, 2018.
Article in Chinese | WPRIM | ID: wpr-687274

ABSTRACT

To inverstigate the alkaloids from the twigs and leaves of Ervatamia pandacaqui, eleven known alkaloids were isolated by silica gel, Sephadex LH-20, and ODS column chromatography, as well as RP-HPLC. Their structures were elucidated by UV, IR, MS, and NMR spectral data as coronaridine (1), 3-oxocoronaridine (2), 19S-heyneanine (3), 19R-heyneanine (4), voacangine (5), 3-oxovoacangine (6), voacristine (7), 19-epi-voacristine (8), iso-voacangine (9), coronaridine 7-hydroxyindolenine (10), and voacangine 7-hydroxyindolenine (11). Compounds 1-11 were isolated from E. pandacaqui for the first time.

3.
Journal of Forensic Medicine ; (6): 223-227, 2018.
Article in Chinese | WPRIM | ID: wpr-984926

ABSTRACT

OBJECTIVES@#To analyse the Fourier transform infrared (FTIR) spectral data of renal tissue at different temperatures in rats after death, and to explore the effects of temperature on the FTIR spectral characteristics of renal tissue.@*METHODS@#The rats were sacrificed by cervical dislocation and placed at 4 ℃, 20 ℃ and 30 ℃. The FTIR spectral data of renal tissue were collected at different time points and analysed by data mining method.@*RESULTS@#The principal component analysis (PCA) results showed that there were significant trends of clustering in the samples of partial time point at 4 ℃, 20 ℃ and 30 ℃. Partial least square (PLS) regression models were established with the spectral data at three temperature groups. The performance of PLS regression models in 20 ℃ and 30 ℃ groups were more superior than that in 4 ℃ group, and the stability of the model in 20 ℃ group was better than that in 30 ℃ group.@*CONCLUSIONS@#There are differences in the FTIR spectral characteristics of renal tissue of rats after death at different temperatures. Temperature has a major impact on the performance of FTIR spectral PLS regression model. Therefore, in order to improve the accuracy of postmortem interval estimation, the effects of temperature on the model should be considered in the related study by spectral method.


Subject(s)
Animals , Rats , Autopsy , Death , Postmortem Changes , Spectroscopy, Fourier Transform Infrared/methods , Temperature
4.
International Eye Science ; (12): 1898-1900, 2017.
Article in Chinese | WPRIM | ID: wpr-641083

ABSTRACT

AIM: To study the clinical effects of using sodium hyaluronate in the glaucoma trabeculectomy. ·METHODS: Totally 46 patients were included in our research and they were divided into two groups. In the study group (23 cases 23 eyes), we used slice with MMC of 0. 4mg/mL in trabeculectomy. Before the conjunctiva flap was sewed, a small amount of sodium hyaluronate was injected into the space under scleral flap and the anterior chamber. In control group ( 23 cases 23 eyes ) , we only used MMC. After trabeculetomy, shallow anterior chambe formation, intraocular pressure, corneal endothelial injury and the formation of filter-bubble were observed in long term. ·RESULTS:IOP of study group at 1d after operation was higher than that of control group ( P0. 05). At 3d after operation, visual acuity decreased in 2 eyes (9%) of study group, 8 eyes (35%) in control group with significant different ( P = 0. 035 ). At 6mo after operation, visual acuity decreased in 0 of study group, 5 eyes (22%) in control group with significant different (P=0. 025 ). The differences on the occurrence of shallow anterior chamber at 7d after operation, formation of filter-bubble at 6mo after operation and corneal endothelial count were significant (P<0. 05). There were less postoperative complications in study group than in control group. ·CONCLUSION: Using sodium hyaluronate combined with MMC in trabeculectomy can greatly prevent the formation of shallow anterior chamber, filter-bubble scarring, and corneal endothelium dropout, make it possible to a safer surgery and a better outcome.

5.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 243-248, 2016.
Article in English | WPRIM | ID: wpr-285279

ABSTRACT

Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.


Subject(s)
Humans , Cell Movement , Genetics , Cytoskeletal Proteins , Genetics , Metabolism , Cytoskeleton , Metabolism , Human Umbilical Vein Endothelial Cells , Cell Biology , Metabolism , Physiology , Neovascularization, Physiologic , Genetics
6.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 243-8, 2016.
Article in English | WPRIM | ID: wpr-638110

ABSTRACT

Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.

7.
Journal of Zhejiang University. Medical sciences ; (6): 466-473, 2014.
Article in Chinese | WPRIM | ID: wpr-251679

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the association of 1858C/T polymorphism of protein tyrosine phosphatase nonreceptor type 22 (PTPN22) and rheumatoid arthritis (RA) susceptibility.</p><p><b>METHODS</b>CMB, wanfang (Chinese) and PubMed databases were searched to get the studies on the association between 1858C/T polymorphism and RA susceptibility, and odds ratio (OR) and 95% confidential interval (CI) were calculated under different genetic models. Then heterogeneity, stratified analysis, and publication bias test were conducted in the study.</p><p><b>RESULTS</b>A total of 32 studies (40 separate comparisons) with 25 059 RA patients and 25 466 controls were included in this meta-analysis. No evidence for publication bias was found in these studies. Meta-analysis showed an association between PTPN22 1858C/T polymorphism and RA (OR=1.606, 95%CI: 1.518-1.699, P<0.001). When stratified by ethnicity, T allele of PTPN22 1858C/T polymorphism was a risk allele in Caucasian (OR=1.612, 95%CI: 1.544-1.683, P<0.001); however, the polymorphism was not detected in Asians (or allele frequencies was extremely low). PTPN22 1858C/T polymorphism was associated with rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibody (ACCP).</p><p><b>CONCLUSION</b>T allele of PTPN22 1858C/T polymorphism is associated with RA susaptibility in Caucasians. PTPN22 1858C/T polymorphism is significantly more prevalent in RF-positive or ACCP-positive patients than in RF-negative or ACCP-negative patients.</p>


Subject(s)
Humans , Arthritis, Rheumatoid , Genetics , White People , Genetics , Genetic Predisposition to Disease , Polymorphism, Single Nucleotide , Protein Tyrosine Phosphatase, Non-Receptor Type 22 , Genetics
8.
Chinese Journal of Applied Clinical Pediatrics ; (24): 902-904, 2013.
Article in Chinese | WPRIM | ID: wpr-733072

ABSTRACT

Objective To investigate the life quality status in autistic disorder children and its influencing factors.Methods Pediatric Quality of Life Inventory (PedsQL4.0) was used to measure the life quality of 200 children with autistic disorder and 120 healthy children.Application of Gesell Development Diagnosis Scale (1981) test was used for intelligence development.Clancy Autism Behavior Scale,Autism Behavior Checklist,and Childhood Autism Rating Scale were used to evaluate the illness degree.Results The life quality of autistic disorder group was lower than that of control group in the scores of physical functioning,emotional functioning,social functioning,mental domain and the totals cores of PedsQL,the differences were significant (all P <0.01).High-functioning autism in all aspects and the overall life quality were higher than those of low function group,the differences were statistically significant (all P <0.01).By using variance analysis,intelligence factors had an impact on the patient's life quality,the difference was statistically significant(P < 0.01).Correlation analysis results suggested that the autism condition degree was heavier,the intelligence level was lower,the life quality was worse,and the difference was statistically significant (P < 0.05).Conclusion Children with autistic disorder suffer grievous influence in life quality.

9.
Acta Physiologica Sinica ; (6): 158-164, 2013.
Article in Chinese | WPRIM | ID: wpr-333121

ABSTRACT

The aim of the present study is to observe the dynamic changes of γ-tubulin in mouse somatic nuclear transferred (SCNT) embryos. The γ-tubulin was detected and analyzed in the enucleated oocyte and SCNT embryos by immunofluorescence and laser confocal microscopy. The results showed that γ-tubulin distributed in the cortex of the enucleated MII oocytes, and decreased in this area during the activation of oocytes. Meanwhile cytoplasmic asters appeared, but there was no spindle formed. Spindle formation could be observed in the enucleated oocytes which were injected with cumulus cells and activated by SrCl2. The spots-like γ-tubulin signals spread between chromosomes at the prophase, and the signals arrayed with spindle or aggregated at two poles of the spindle at the early metaphase. Furthermore, γ-tubulin signals were localized between the segregated sister chromatids at anaphase or telophase. Some reconstructed embryos exhibited advanced activation, showing abnormal spindles and aberrant distribution of γ-tubulin and chromosomes. Two spindles would be formed when the cumulus cell was injected into an intact oocyte, and the distribution of γ-tubulin was similar to that of the normal SCNT. Moreover, advanced activation also occurred in this case and formed either two small spindles or one big barrel-shaped spindle. These results suggest that γ-tubulin plays a pivotal role in spindle assembling in mouse SCNT embryos. The reconstructed oocytes were easily to be activated, and aberrant distribution of γ-tubulin is associated with formation of abnormal spindles and chromosome misalignment.


Subject(s)
Animals , Mice , Embryo, Mammalian , Metabolism , Fluorescent Antibody Technique , Metaphase , Microscopy, Confocal , Nuclear Transfer Techniques , Oocytes , Cell Biology , Spindle Apparatus , Metabolism , Telophase , Tubulin , Metabolism
10.
Acta Academiae Medicinae Sinicae ; (6): 51-54, 2010.
Article in Chinese | WPRIM | ID: wpr-301595

ABSTRACT

<p><b>OBJECTIVE</b>To validate and supplement the libraries of serial analysis of gene expression (SAGE) by the application of the real-time quantitative polymerase chain reaction PCR).</p><p><b>METHODS</b>The primers were designed based on the full sequences of the genes. Nine single matched tags, 6 multiple matched tags, 1 non-matched tag due to the update of the National Center for Biotechnology Information (NCBI) database, and 2 non-matched tags were selected to fulfill the validation of real-time PCR.</p><p><b>RESULTS</b>The genes were all specifically amplified by the primers pairs. The expressions of the single matched tags were identical to those of the SAGE libraries; however, the expressions of only 3 genes of the 6 multi-matched tags were identical to those of the SAGE libraries. The PCR data of the non-matched tag due to the update of the NCBI database were opposite to those of the SAGE libraries. The data did not support the significant difference of the non-matched gene of the SAGE libraries.</p><p><b>CONCLUSIONS</b>Real-time PCR is a reliable tool for the validation of high through-put data such as SAGE. The reliability of data depends on the match of the tags of the SAGE libraries.</p>


Subject(s)
Gene Expression Profiling , Methods , Oligonucleotide Array Sequence Analysis , Methods , Real-Time Polymerase Chain Reaction
11.
Journal of Southern Medical University ; (12): 195-198, 2009.
Article in Chinese | WPRIM | ID: wpr-339033

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of angiotensin II (Ang II) on pregnancy-associated plasma protein A (PAPP-A) and insulin-like growth factor 1 (IGF-1) mRNA expressions in human umbilical artery smooth muscle cells (HUVSMCs).</p><p><b>METHODS</b>In the presence or absence of Ox-LDL, HUVSMCs were cultured with Ang II of 10(-5) mol/L for 0, 6, 12, 24, 36, and 48 h, or with Ang II at the concentrations of 10(-7), 10(-6), 10(-5), and 10(-4) mol/L for 24 h, after which the cells were then collected to detect PAPP-A and IGF-1 mRNA expressions in the cells using RT-PCR.</p><p><b>RESULTS</b>At the concentration of 10(-5) mol/L, Ang II showed a time-dependent effect in inducing PAPP-A and IGF-1 mRNA expressions, which began to increase at 12 h of culture and reaching the highest level at 24 h. Ang II also dose-dependently induced PAPP-A and IGF-1 mRNA expressions, and 10(-5) mol/L Ang II induced the highest expression levels of the two genes. Ox-LDL exposure significantly further increased the expression levels of PAPP-A and IGF-1 mRNA in the cells regardless of the Ang II concentration or duration for cell treatment (P<0.05).</p><p><b>CONCLUSION</b>Ang II can time- and dose-dependently induces PAPP-A and IGF-1 mRNA expression in HUVSMCs and is responsible for inducing platelet activity and inflammatory reaction in acute coronary syndromes, and the effects of Ang II can be enhanced by Ox-LDL.</p>


Subject(s)
Humans , Angiotensin II , Pharmacology , Cells, Cultured , Drug Synergism , Insulin-Like Growth Factor I , Genetics , Metabolism , Lipoproteins, LDL , Pharmacology , Myocytes, Smooth Muscle , Cell Biology , Metabolism , Pregnancy-Associated Plasma Protein-A , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Umbilical Arteries , Cell Biology , Metabolism
12.
Chinese Journal of Gastrointestinal Surgery ; (12): 52-56, 2009.
Article in Chinese | WPRIM | ID: wpr-326558

ABSTRACT

<p><b>OBJECTIVE</b>To screen differential expression genes and proteins at transcriptome and proteome levels between human gastric cancer tissue and corresponding normal mucosa.</p><p><b>METHODS</b>Fresh-frozen gastric cancers were collected from patients treated at Ruijin Hospital. A total of 22 pairs of gastric cancer tissues and the corresponding noncancerous mucosa were analyzed. Commercially available cDNA microarray with 14 592 genes/ESTs was used. Genes were considered to be up-or down-regulated when the intensity ratio Cy3/Cy5 was > or = 2 or < or = 0.5 in over 50% samples (P<0.05). Immobilized pH gradient(IPG)-based 2-DE was applied to separate the total proteins of gastric cancer tissue and paired normal tissue. After staining and analysis by software,the differential expression proteins were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) or MALDI-TOF-TOF-MS.</p><p><b>RESULTS</b>As compared with corresponding noncancerous tissue, there were totally 149 up-regulating genes/ESTs and 238 down-regulating genes/ESTs in gastric cancer, including 29 genes with 3-fold over-expression ratio and 21 genes with 5-fold under-expression. Fifteen protein spots were identified successfully, among whom there were ten over-expressed and five under-expressed proteins in gastric cancer tissue compared with normal tissue. Most of over-expressed genes and proteins were related to cell motility, cell proliferation, signal transduction, while those under-expressed genes and proteins were related to defense response, toxoid metabolism.</p><p><b>CONCLUSION</b>Studying gastric cancer at transcriptome and proteome levels can help demonstrate tumorigenesis and biological characteristics of gastric cancer comprehensively and provide powerful tools to find new biomarkers associated with gastric cancer and therapy targets.</p>


Subject(s)
Humans , Electrophoresis, Gel, Two-Dimensional , Expressed Sequence Tags , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Genes, Neoplasm , Genome, Human , Neoplasm Proteins , Neoplasm Staging , Oligonucleotide Array Sequence Analysis , Proteome , Metabolism , Proteomics , Stomach Neoplasms , Genetics , Metabolism , Pathology
13.
Acta Physiologica Sinica ; (6): 777-782, 2008.
Article in Chinese | WPRIM | ID: wpr-302491

ABSTRACT

Tubulin is the major protein of microtubule. alpha- and beta- tubulins form heterodimers, while gamma-tubulin regulates microtubule organization. The present study aimed to observe the dynamic changes of gamma-tubulin in preimplantation development of parthenogenetic mouse embryos. Immunofluorescence and laser confocal microscopy were used to detect the location of gamma-tubulin in preimplantation parthenogenetic embryos activated by SrCl2. The oocytes were collected at 13-14 h after hCG injection, and then activated with 10 mmol/L SrCl2 in Ca(2+)-free CZB medium with 5 mmol/L cytochalasin B (CB), fixed at 1 h intervals until 6 h after activation. The results showed that spindle was paralleled with the cell membrane all the time, when the meiosis of MII mouse oocytes resumed. The rotation of spindle was inhibited, but karyokinesis was not influenced. At 0 h after activation, i.e. at metaphase, gamma-tubulin was distributed mainly on the two poles of spindle. At 1 h after activation, i.e. at anaphase, following the separation of chromosomes, gamma-tubulin was transformed from dense to disperse. At 2 h after activation, gamma-tubulin was localized between the segregated sister chromatids at telophase. However, at 3-6 h after activation, gamma-tubulin concentrated around the two female pronuclei during their formation and juxtaposition. Moreover, another group of MII oocytes were activated for 6 h and cultured in droplets of KSOM medium under mineral oil in 5% CO2 in air at 37 degrees C to permit parthenogenetic development. The embryos were collected and fixed at 3 h, 14 h, 16 h, and 18 h of culture. At 3 h after culture, i.e. at mitotic interphase, it was shown that amorphous gamma-tubulin distributed around the nuclei of early parthenogenetic embryos. At 24 h after culture, i.e. at prometaphase, gamma-tubulin migrated along the spindle microtubule to the two poles. Our results showed that gamma-tubulin had similar location patterns at metaphase, anaphase and telophase in meiosis and mitosis. It was concluded that gamma-tubulin assembly in parthenogenetically activated oocytes facilitated the formation of negative pole cap and the stabilization of microtubule, thus promoting the spindle formation at meiosis and mitosis. The relocation of gamma-tubulin at anaphase and telophase might be induced by the event of segregation of homologous chromosome being pulled away by the spindle. gamma-tubulin might contribute to the migration and juxtaposition of the two female pronuclei as well.


Subject(s)
Animals , Female , Mice , Embryo, Mammalian , Embryonic Development , Meiosis , Mitosis , Oocytes , Cell Biology , Parthenogenesis , Spindle Apparatus , Physiology , Tubulin , Physiology
14.
Microbiology ; (12)2008.
Article in Chinese | WPRIM | ID: wpr-686316

ABSTRACT

According to morphological and microscopic characteristics, a high-efficient decolorizing fungus, the strain Asaw117, was identified as Aspergillus awamori. Selecting eight different dyes from Azo dyes, anthraquinones dyes and oxygen Quinones dyes, the decolorizing assays of various dyes showed that the strain Asaw 117 was the highest decolorizing potential to 0.1 g/L Vat Blue RSN, the discoloration rate up to 100 percent. Comparing to different kinds of medium and several of carbon and nitrogen sources, the strain had the best decolorizing efficient although grew slower in the Czapek medium, otherwise, grew quicker and decolorizing efficient lower in the PDF medium. It could use Vat Blue RSN as a nitrogen source, but not as a carbon source. The medium composing of saccharose and ammonium nitrate as carbon and nitrogen sources was decolorizing potential markedly during different combinations of carbon and nitrogen sources. So the strain has good potential for the dyeing wastewater treatment

15.
Chinese Journal of Gastrointestinal Surgery ; (12): 212-216, 2007.
Article in Chinese | WPRIM | ID: wpr-336473

ABSTRACT

<p><b>OBJECTIVE</b>To investigate relationship between the expression of hepatocyte growth factor(HGF) and lymphatic vessel formation in gastric cancer(GC), and to further clarify the possible ways of HGF promoting GC lymphangiogenesis and lymphatic metastasis.</p><p><b>METHODS</b>The expression of HGF, VEGF-C, c-Met and microlymphatic density(MLD) in tumor and non-tumor areas of 60 cases of GC and 20 cases of peptic ulcer (PU) were evaluated by immunohistochemistry. The relationships between MLD and clinicopathologic features of GC were analyzed.</p><p><b>RESULTS</b>The proteins of HGF, VEGF-C, and c-Met were overexpressed in GC tissues, which were significantly higher than those in normal gastric and PU tissues (P<0.001). The MLD of GC was also significant higher than those of normal gastric tissue and PU tissues (P<0.05). C-Met was expressed on enlarge lymphatic vessels in GC and PU tissues. MLD was positively correlated with the expression of HGF in GC (P<0.001).</p><p><b>CONCLUSION</b>HGF as well as VEGF-C is a lymphangiogenic factor in GC. HGF can directly or indirectly stimulate lymphangiogenesis and contribute to lymphatic metastasis in GC.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Hepatocyte Growth Factor , Metabolism , Lymphangiogenesis , Lymphatic Metastasis , Lymphatic Vessels , Pathology , Proto-Oncogene Proteins c-met , Metabolism , Stomach Neoplasms , Metabolism , Pathology , Vascular Endothelial Growth Factor C , Metabolism
16.
Chinese Journal of Biotechnology ; (12): 1116-1120, 2007.
Article in Chinese | WPRIM | ID: wpr-276151

ABSTRACT

<p><b>UNLABELLED</b>To reduce the huge labor-cost in the screening in traditional monoclonal antibody generation, We established a new system for monoclonal antibody generation integrating with protein array. BALB/c mice were immunized by eight recombinant proteins respectively, and the positive hybridoma cells were obtained by cell fusion and ELISA screening. All the eight kinds of positive hybridoma cells were mixed, cloned, screened by protein array, and definite dilution cloned.</p><p><b>RESULTS</b>175 single cell clones were obtained by complex cloning, and 119 of those were positive clones. Then 8 positive cell lines were generated by the following 2 rounds definite dilution cloning. By comparing with the traditional method, we got 8 monoclonal antibodies using the combined protein array screening and multiplex cloning method in 1 cycle, and fewer amounts of antigens were used. As a result, the combined protein array and multiplex cloning method could be used as an economical, rapid and simple tool applying in high throughput monoclonal antibody generation.</p>


Subject(s)
Animals , Female , Humans , Mice , Antibodies, Monoclonal , Allergy and Immunology , Hybridomas , Metabolism , Mice, Inbred BALB C , Protein Array Analysis
17.
Chinese Journal of Oncology ; (12): 96-100, 2007.
Article in Chinese | WPRIM | ID: wpr-255714

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of hypoxia and hypoxia-inducible factor-1alpha (HIF-1alpha) on the expression of multidrug resistance gene-1 (mdr-1) and its coded p-glyeoprotein (P-gp) as well as the chemotherapeutic sensitivity of human ovarian cancer cells to paclitaxel and its mechanism.</p><p><b>METHODS</b>The mRNA and protein levels of HIF-1alpha, mdr-1 and p-gp were studied by immunocytochemistry, semiquantitative reverse transcription-ploymerase chain reaction (RT-PCR) and Western blot analysis in human ovarian cancer cells (A2780) in 5% CO2 + 1% O2 hypoxic culture and 21% O2 normoxic culture, respectively. Methyl thiazolyl tetrazolium (MIT) was used to evaluate the chemotherapeutic sensitivity of A2780 cells to paclitaxel by inhibition rate. RNA interference technique was used and small hairpin RNAs (shRNAs) eukaryotic expression vector targeting HIF-1alpha was constructed as pSiHIF-1alpha, and transfected into A2780 cells. RT-PCR and Western blot were used to detect gene silencing effect on HIF-1alpha, the expressions of mdr-1 and p-gp. The inhibition rate was observed after HIF-1alpha gene silence.</p><p><b>RESULTS</b>HIF-1alpha at both mRNA and protein levels was induced significantly under hypoxia. The HIF-1alpha expression at mRNA level was oxygen gradient-independent, while HIF-1alpha expression at protein level was oxygen gradient-dependent. The inhibition rate of paclitaxel to hypoxic A2780 cells in 5% CO2 + 1% O2 was significantly lower than that in normoxic A2780 cells (P <0.05). The shRNAs plasmid targeting HIF-1alpha was constructed successfully and HIF-1alpha gene was silenced in A2780 cells efficiently followed by mdr-1 and p-gp down-regulation. The inhibition rate was greatly increased in hypoxic A2780/siHIF-1alpha cells.</p><p><b>CONCLUSION</b>Hypoxia can decrease the chemotherapeutic sensitivity of human ovarian cancer A2780 cells to paclitaxel through HIF-1alpha regulating the expression of mdr-1 and p-gp.</p>


Subject(s)
Female , Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Genetics , Antineoplastic Agents, Phytogenic , Pharmacology , Blotting, Western , Cell Hypoxia , Cell Line, Tumor , Cell Survival , Dose-Response Relationship, Drug , Down-Regulation , Drug Resistance, Neoplasm , Genetics , Hypoxia-Inducible Factor 1, alpha Subunit , Genetics , Ovarian Neoplasms , Genetics , Metabolism , Pathology , Paclitaxel , Pharmacology , RNA Interference , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Transfection
18.
Chinese Journal of Oncology ; (12): 116-119, 2006.
Article in Chinese | WPRIM | ID: wpr-308406

ABSTRACT

<p><b>OBJECTIVE</b>To identify cancer-related genes in diffuse-type gastric cancer and to explore its molecular mechanism by cDNA microarray analysis.</p><p><b>METHODS</b>A total of 22 pairs of diffuse-type gastric cancer tissue and the corresponding normal mucosa were taken and freshly frozen. cDNA microarray with 14,592 genes/ESTs was used. Genes were considered to be up- or down-regulated when the fluorescent intensity ratio between tumor and normal mucosa was over 2-fold in over 50% of the samples (P < 0.05). Hierarchical clustering of regulated genes was performed as a measure to study expressional similarity. Validation of array results was carried out by real time quantitative PCR (QPCR).</p><p><b>RESULTS</b>Compared with those of corresponding normal mucosa, there were a total of 153 genes/ESTs up-regulated and 204 down-regulated in diffuse-type gastric cancer. Hierarchical clustering demonstrated that the genes belonging to the same subgroup displayed similar function. Most of the overexpressed genes were those related to cell adhesion, cell motility, matrix reconstruction, cell proliferation and/or signal transduction; while genes related to defense response, toxicoid metabolism, DNA repairing, nuclear-cytoplasmic transport and/or anti-apoptosis made up the main list of the underexpressed genes. Seven genes showed higher expression in TNM (T I + T II) group than in (T III + T IV) group. QPCR confirmed the array analysis results.</p><p><b>CONCLUSION</b>Gene expression profiling by cDNA microarray analysis provides not only molecular understanding of biological properties of cancer, but may also be helpful in discovering new diagnostic markers and therapeutic targets in gastric adenocarcinoma.</p>


Subject(s)
Female , Humans , Male , Middle Aged , Adenocarcinoma , Genetics , Metabolism , Biglycan , Collagen Type I , Metabolism , Expressed Sequence Tags , Extracellular Matrix Proteins , Metabolism , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Microfilament Proteins , Metabolism , Muscle Proteins , Metabolism , Neoplasm Staging , Oligonucleotide Array Sequence Analysis , Pepsinogen C , Metabolism , Proteoglycans , Metabolism , Stomach Neoplasms , Genetics , Metabolism
19.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 425-426, 2004.
Article in Chinese | WPRIM | ID: wpr-979101

ABSTRACT

@#Objective To observe the therapeutic effectiveness of monosialo gangliosides injection (GM-1) on cerebral palsy (CP) of children.Methods 87 children with CP were randomly divided into the observation group (51 cases) and control group (36 cases), and treated with physical therapy, acupuncture, massage and hyperbaric oxygen, but monosialo gangliosides was added to the observation group only. The development quotient (DQ) of each group was calculated by Gesell development schedules before and after the therapy respectively. Therapeutic effects of two groups were compared.Results The therapeutic effect of the observation group was more satisfactory than that of control group(P<0.05). The observation group expressed higher DQ and scores of gross motor, fine motor, language and reaction than that of control group. Conclusion Monosialo ganglioside is effective on CP.

20.
Chinese Journal of Hematology ; (12): 266-268, 2004.
Article in Chinese | WPRIM | ID: wpr-291452

ABSTRACT

<p><b>OBJECTIVE</b>To better understand the mechanisms of the fetal hematopoiesis turn over from primitive to definitive hematopoiesis through the expression level of c-kit(+) and sca-1(+), and major characters of gene expression profile of these cells.</p><p><b>METHODS</b>c-kit and sca-1 expression level were monitored with fluorescence activated cell sorting (FACS) of the mononuclear cells from mouse yolk sac and fetal liver, while gene expression profile was carried out with EST sequencing strategy.</p><p><b>RESULTS</b>The Sca-1(+) cells were increased while the c-kit(+) cells decreased with the embryonic development. Through profiling the functionally identified known genes, most of the highly expressed were globin genes, especially of embryonic types.</p><p><b>CONCLUSION</b>The erythropoiesis played a key role in early fetal hematopoiesis in mammalian.</p>


Subject(s)
Animals , Mice , Antigens, Ly , Genetics , Metabolism , Cell Differentiation , Genetics , Flow Cytometry , Gene Expression Profiling , Gene Expression Regulation, Developmental , Liver , Cell Biology , Embryology , Metabolism , Membrane Proteins , Genetics , Metabolism , Mice, Inbred C57BL , Proto-Oncogene Proteins c-kit , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Yolk Sac , Cell Biology , Embryology , Metabolism
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