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1.
Journal of Medical Postgraduates ; (12): 523-525, 2019.
Article in Chinese | WPRIM | ID: wpr-818272

ABSTRACT

Objective Reports are rarely seen on the clinical studies of pituicytomas. This article discusses the clinical diagnosis, treatment strategies, pathological features and postoperative complications of pituicytomas. Methods This retrospective study included 10 cases of pituicytomas treated in our hospital from July 2008 to July 2018. All the patients underwent microsurgery, gross total resection of the tumor in 6 cases and subtotal resection in the other 4. We analyzed the histopathological features, postoperative complications and follow-up data of the patients. Results The tumors were tightly organized morphologically, consisting of bipolar spindle-shaped cells, arranged like crosswise-woven fiber bundles or in a storiform pattern, the cytoplasm stained red and eosinophilic, the nuclei medium-sized, round, oval or spindle shaped, mildly heterotypical, with visible microencapsulation and abundant interstitial blood vessels. The main postoperative complications included insipidus in 8 cases, hyponatremia in 4, decreased visual acuity in 3, hypopituitarism in 2, and intracranial infection in 1. Of the 9 patients followed up, 1 experienced recurrence at 2 years postoperatively and received another surgery, with no more recurrence hitherto. Conclusion Definite diagnosis of pituicytoma depends on pathological examination, and adequate attention should be paid to the prevention and management of such postoperative complications as insipidus and hyponatremia.

2.
Acta Physiologica Sinica ; (6): 588-596, 2019.
Article in Chinese | WPRIM | ID: wpr-777152

ABSTRACT

The aim of the study was to establish Ace2 (angiotensin-converting enzyme 2) knockout mouse model with CRISPR/Cas9 gene targeting technology. A vector targeting Ace2 gene knockout was constructed with the primers of single-guide RNA (gRNA), and then transcribed gRNA/Cas9 mRNA was micro-injected into the mouse zygote. The deletion of exons 3 to 18 of Ace2 gene in mice was detected and identified by PCR and gene sequencing. The Ace2 gene knock-out mice were bred and copulated. Ace2 protein and mRNA expression were detected by Western blot and qRT-PCR in F3 progeny knock-out male mice. The gRNA expression vector was successfully constructed and transcribed in vitro, and active gRNA and Cas9 mRNA were injected directly into zygote. The deletion of exons 3 to 18 of Ace2 gene in six positive founder mice as the F0 generation were confirmed by PCR and gene sequencing. Six founder mice were mated with wild-type mice, then achieved F1 generation were mated and produced F2 generation. The female positive mouse of F2 was selected to mate with wild-type mice and produce Ace2 mice of F3 generation. Ace2 mRNA and protein were not detected in tissues of these Ace2 mice. In conclusion, a mouse model with Ace2 deficiency has been successfully established with CRISPR/Cas9 technique, which shall lay a foundation for future investigation of Ace2.


Subject(s)
Animals , Female , Male , Mice , CRISPR-Cas Systems , Gene Knockout Techniques , Gene Targeting , Mice, Knockout , Genetics
3.
Chinese Journal of Immunology ; (12): 15-18,24, 2018.
Article in Chinese | WPRIM | ID: wpr-702665

ABSTRACT

Objective:To study the influences of Mycoplasma penumoniae capsular polysaccharide (CPS) on the phagocytosis and membrane molecules expression of the human peripheral blood mononuclear cells derived dendritic cells by binding to dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN),so as to know the effect of CPS on the maturation of dendritic cells.Methods:M.pneumoniae strain was cultivated and CPS was extracted.Human peripheral blood mononuclear cells were separated and induced to dendritic cells,then identified the cells by flow cytometry and observation under the microscope.CPS was used to treat dendritic cells or cells pretreated with DC-SIGN monoclonal antibody,and then FITC-dextran phagocytosis and surface markers CD83,HLA-DR,CD80 and CD86 were detected by flow cytometry.Results:The dendritic cells tended to form colony groups.The positive rate of CD11c molecule in the cultured dendritic cells was about 86.27%.After stimulated by CPS,the FITC-dextran fluorescence mean intake by dendritic cells were increased (P<0.05),while the cell surface membrane molecules CD83,HLA-DR,CD80 and CD86 were decreased significantly when compared with the PBS treated control cells(P<0.05).When blocked DC-SIGN with the monoclonal antibody,the FITC-dextran fluorescence mean and membrane molecules expression had no statistical difference with the control cells(P>0.05).Conclusion:M.pneumoniae CPS can promote the phagocytic function of DC and inhibit the expression of CD83,HLA-DR,CD80 and CD86.

4.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1459-1462, 2015.
Article in Chinese | WPRIM | ID: wpr-286361

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of Jinying Capsule (JC) in treating pelvic inflammatory disease patients with accumulated damp-heat syndrome (ADHS).</p><p><b>METHODS</b>Totally 328 patients were recruited in a prospective, positive drug parallel controlled, and multi-center clinical trial. Of them 213 patients in the treatment group took JC (0.5 g per capsule), 4 capsules each time, 3 times per day, while 115 patients in the control group took Kangfuyan Capsule (KC, 0.4 g per capsule), 3 capsules each time, twice per day. The course of treatment was 4 weeks for all. Scores of Chinese medical syndromes, visual analogue scale (VAS) of the lower abdominal pain, and European quality of life-five dimension scale (EQ-5D) were observed before treatment and after 4 weeks of treatment.</p><p><b>RESULTS</b>There were 204 patients in the treatment group and 109 in the control group who completed this trial. The total effective rate of Chinese medical syndrome was 89.71% (183/204 cases) in the treatment group and 76.15% (83/109 cases) in the control group (P < 0.01). Compared with before treatment in the same group, EQ-5D scores increased, and VAS scores of the lower abdominal pain decreased in the two groups after treatment. EQ-5D scores was 0.857 ± 0.157 in the treatment group, obviously higher than that in the control group (0.753 ± 0.126, P < 0.05). VAS scores of the lower abdominal pain was 2.14 ± 1.23 in the treatment group, lower than that in the control group (2.33 ± 1.24), but with no statistical difference between the two groups (P > 0.05). No adverse reaction occurred in the two groups.</p><p><b>CONCLUSION</b>JC was superior to KC in improving Chinese medical syndrome and quality of life of pelvic inflammatory disease patients with accumulated damp-heat syndrome.</p>


Subject(s)
Female , Humans , Capsules , Drugs, Chinese Herbal , Therapeutic Uses , Hot Temperature , Medicine, Chinese Traditional , Pelvic Inflammatory Disease , Drug Therapy , Phytotherapy , Prospective Studies , Quality of Life , Safety , Syndrome
5.
Chinese Journal of Contemporary Pediatrics ; (12): 395-399, 2015.
Article in Chinese | WPRIM | ID: wpr-346140

ABSTRACT

<p><b>OBJECTIVE</b>To study the protocol of construction of the mutation E292V and M723I of hABCA3 gene associated with neonatal respiratory distress syndrome, as well as their eukaryotic green fluorescent protein expression rectors, and to examine the expression of mutation proteins in human lung carcinoma epithelial cells (A549).</p><p><b>METHODS</b>Site-directed mutagenesis method based on overlap extension PCR was used to introduce mutations in the two sites which were E292V and M723I in the ABCA3. The PCR fragments were subcloned to PEGFP-C2 vectors to construct the eukaryotic green fluorescent protein expression rectors. A549 cells were transiently transfected with the recombinants using Lipofectamine 2000 and the transfection efficiency was confirmed through GFP signal. The expression and location of recombinants were detected by FV1000 laser scanning microscope.</p><p><b>RESULTS</b>Direct sequence analysis confirmed an A to T transition at position 875 in E292V and a G to A transition at position 2169 in M723I. Recombinants were transfected to A549 cells and both wild type and mutant ABCA3 proteins were expressed in the cytoplasm.</p><p><b>CONCLUSIONS</b>The eukaryotic green fluorescent protein expression rectors of wild type and mutant ABCA3 gene were constructed and they were successfully expressed in A549 cells. This experiment provides a basis for subsequent research.</p>


Subject(s)
Humans , ATP-Binding Cassette Transporters , Genetics , Cell Line, Tumor , Green Fluorescent Proteins , Genetics , Mutagenesis, Site-Directed , Transfection
6.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 750-4, 2014.
Article in English | WPRIM | ID: wpr-636744

ABSTRACT

The influence of inner cell mass (ICM) and trophectoderm (TE) score on pregnancy outcomes in frozen-thawed blastocyst transfer cycles was analyzed. A retrospective analysis of 741 cycles of frozen-thawed blastosysts transfer was performed. All cycles were divided into four groups based on the number and morphological score of blastocysts: S-ICM B/TE B group (n=91), the single blastocyst transfer of ICM B and TE B; D-ICM B/TE B group (n=579), double blastocysts transfer of ICM B/TE B; D-ICM B/TE C group (n=35), double blastocysts transfer of ICM B/TE C; and D-ICM C/TE B group (n=36), double blastocysts transfer of TE B/ICM C. The pregnancy outcomes were compared among the four groups. As compared with D-ICM B/TE C group, the clinical pregnancy rate, implantation rate and multiple pregnancy rate were increased in D-ICM B/TE B group (74.96% vs. 57.14%, 57.43% vs. 37.14%, and 48.62% vs. 25%, respectively, P<0.05 for all). Clinical pregnancy rate and implantation rate in D-ICM B/TE B group were also higher than in D-ICM C/TE B group (74.96% vs. 50%, and 57.43% vs. 33.33%, both P<0.05). Multivariable Logistic regression analysis indicated that ICM score was a better predictive parameter for clinical pregnancy (OR=3.05, CI 1.70-5.46, P<0.001), while the trophectoderm score was a better one for early abortion (OR=0.074, CI 0.03-0.19, P<0.001). Clinical pregnancy rate and multiple pregnancy rate in S-ICM B/TE B group were significantly lower than those in D-ICM B/TE B group (46.15% vs. 74.96%, and 2.38% vs. 48.62%, both P<0.05), but there was no significant difference in the implantation rate between the two groups. It was suggested that the higher score of ICM and TE may be indicative of the better pregnancy outcomes. The ICM score is a better predictor of clinical pregnancy than TE, while TE score is a better one in predicting early abortion. Single ICM B/TE B blastocyst transfer in frozen-thawed cycles can also get satisfactory pregnancy outcomes.

7.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 750-754, 2014.
Article in English | WPRIM | ID: wpr-331150

ABSTRACT

The influence of inner cell mass (ICM) and trophectoderm (TE) score on pregnancy outcomes in frozen-thawed blastocyst transfer cycles was analyzed. A retrospective analysis of 741 cycles of frozen-thawed blastosysts transfer was performed. All cycles were divided into four groups based on the number and morphological score of blastocysts: S-ICM B/TE B group (n=91), the single blastocyst transfer of ICM B and TE B; D-ICM B/TE B group (n=579), double blastocysts transfer of ICM B/TE B; D-ICM B/TE C group (n=35), double blastocysts transfer of ICM B/TE C; and D-ICM C/TE B group (n=36), double blastocysts transfer of TE B/ICM C. The pregnancy outcomes were compared among the four groups. As compared with D-ICM B/TE C group, the clinical pregnancy rate, implantation rate and multiple pregnancy rate were increased in D-ICM B/TE B group (74.96% vs. 57.14%, 57.43% vs. 37.14%, and 48.62% vs. 25%, respectively, P<0.05 for all). Clinical pregnancy rate and implantation rate in D-ICM B/TE B group were also higher than in D-ICM C/TE B group (74.96% vs. 50%, and 57.43% vs. 33.33%, both P<0.05). Multivariable Logistic regression analysis indicated that ICM score was a better predictive parameter for clinical pregnancy (OR=3.05, CI 1.70-5.46, P<0.001), while the trophectoderm score was a better one for early abortion (OR=0.074, CI 0.03-0.19, P<0.001). Clinical pregnancy rate and multiple pregnancy rate in S-ICM B/TE B group were significantly lower than those in D-ICM B/TE B group (46.15% vs. 74.96%, and 2.38% vs. 48.62%, both P<0.05), but there was no significant difference in the implantation rate between the two groups. It was suggested that the higher score of ICM and TE may be indicative of the better pregnancy outcomes. The ICM score is a better predictor of clinical pregnancy than TE, while TE score is a better one in predicting early abortion. Single ICM B/TE B blastocyst transfer in frozen-thawed cycles can also get satisfactory pregnancy outcomes.


Subject(s)
Adult , Female , Humans , Pregnancy , Analysis of Variance , Blastocyst , Cell Biology , Blastocyst Inner Cell Mass , Cell Biology , Cryopreservation , Methods , Embryo Implantation , Embryo Transfer , Methods , Fertilization in Vitro , Methods , Pregnancy Outcome , Pregnancy Rate , Retrospective Studies
8.
Chinese Journal of Medical Genetics ; (6): 121-125, 2012.
Article in Chinese | WPRIM | ID: wpr-295524

ABSTRACT

<p><b>OBJECTIVE</b>To screen for potential mutations of LKB1 gene in Chinese familial Peutz-Jeghers syndrome (PJS) patients and analyze their clinical manifestations.</p><p><b>METHODS</b>Eleven PJS families were collected and genomic DNA of peripheral blood was extracted. Typically mucosal pigmentation and hamartomatous polyps were present in all 11 probands. Mutation screening of the probands were carried out by PCR and direct sequencing. Two hundred and fifty healthy adults were enrolled as normal controls, for whom genomic DNA of peripheral blood was also extracted. PCR-denaturing high performance liquid chromatography was carried out to verify the mutation identified in the patients.</p><p><b>RESULTS</b>Nine germline mutations were identified in eight PJS patients, which included 7 point mutations, 1 deletion and 1 insertion. Among these, 4 were considered to be pathogenic, of which 2 were de novel, 4 were considered to be polymorphism, and 1 was uncertain.</p><p><b>CONCLUSION</b>LKB1 gene mutations with pathogenic effect are a common cause of familial PJS in Chinese patients. Most mutations are point mutations.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Base Sequence , China , Genetic Predisposition to Disease , Germ-Line Mutation , Molecular Sequence Data , Peutz-Jeghers Syndrome , Genetics , Protein Serine-Threonine Kinases , Genetics
9.
Chinese Journal of Cancer ; (12): 565-573, 2011.
Article in English | WPRIM | ID: wpr-294489

ABSTRACT

Although many studies have investigated intensity-modulated radiation therapy (IMRT) for nasopharyngeal carcinoma (NPC), sample sizes in the reported studies are usually small and different in outcomes in different T and N subgroups are seldom analyzed. Herein, we evaluated the outcomes of NPC patients treated with IMRT and further explored treatment strategy to improve such outcome. We collected clinical data of 865 NPC patients treated with IMRT alone or in combination with chemotherapy, and classified all cases into the following prognostic categories according to different TNM stages: early stage group (T1-2N0-1M0), advanced local disease group (T3-4N0-1M0), advanced nodal disease group (T1-2N2-3M0), and advanced locoregional disease group (T3-4N2-3M0). The 5-year overall survival (OS), local relapse-free survival (LRFS), and distant metastases-free survival (DMFS) were 83.0%, 90.4%, and 84.0%, respectively. The early disease group had the lowest treatment failure rate, with a 5-year OS of 95.6%. The advanced local disease group and advanced nodal disease group had similar failure pattern and treatment outcomes as well as similar hazard ratios for death (4.230 and 4.625, respectively). The advanced locoregional disease group had the highest incidence of relapse and death, with a 5-year DMFS and OS of 62.3% and 62.2%, respectively, and a hazard ratio for death of 10.402. Comparing with IMRT alone, IMRT in combination with chemotherapy provided no significant benefit to locoregionally advanced NPC. Our results suggest that the decision of treatment strategy for NPC patients should consider combinations of T and N stages, and that IMRT alone for early stage NPC patients can produce satisfactory results. However, for advanced local, nodal, and locoregional disease groups, a combination of chemotherapy and radiotherapy is recommended.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Carcinoma , Chemoradiotherapy , Chemotherapy, Adjuvant , Disease-Free Survival , Lymphatic Metastasis , Nasopharyngeal Neoplasms , Drug Therapy , Pathology , Radiotherapy , Neoplasm Recurrence, Local , Neoplasm Staging , Radiotherapy Dosage , Radiotherapy, Intensity-Modulated , Survival Rate
10.
Chinese Journal of Cancer ; (12): 82-86, 2010.
Article in Chinese | WPRIM | ID: wpr-292635

ABSTRACT

<p><b>BACKGROUND AND OBJECTIVE</b>At present, although appropriate radiotherapy and combined treatments are widely used for the patients with primary nasopharyngeal carcinoma (NPC), local or regional recurrence rates are still high. According to clinical performance, pathology, and diagnostic imaging of the patients with the first recurrence of NPC, this study analyzed the clinical features of recurrent NPC to provide a reference for tracking the rules of recurrence after the treatment of patients with NPC.</p><p><b>METHODS</b>Clinical data of 337 patients diagnosed with recurrent NPC for the first time were collected. The diagnoses were based on pathology and/or imaging and the patients were treated at the Sun Yat-sen University Cancer Center between January 1999 and December 2004. Data used for statistical analysis included clinical performance during the patient visit, the extension of the invasion as shown on imaging, pathologic features, Epstein-Barr virus (EBV) serology, restaging, etc.</p><p><b>RESULTS</b>Patients were staged according to the system developed by the International Union Against Cancer (UICC) and the American Joint Committee on Cancer (AJCC) in 2002. Patients with diseases at stages I/II accounted for 25.2%, while those with stage III/IV accounted for 74.8%. The median interval of relapse was 25 months. Patients had local recurrence (69.4%), regional recurrence (4.5%), or both (26.1%). Epistaxis and headache were the most common symptoms. Abduct dysfunction and facial numbness induced by cranial nerve damage were the most common signs. The probability of invasion of structures adjacent to the nasopharynx, such as the oropharynx, the prestyloid space, and the carotid sheath area, was low in patients with recurrent NPC. By contrast, the probability of invasion of structures far from the nasopharynx, such as the base of the skull, the paranasal sinuses, cranial nerves, the cavernous sinus, the brain, the pterygopalatine fossa, the infratemporal fossa, the orbital apex, and the soft palate, was higher in recurrent NPC.</p><p><b>CONCLUSIONS</b>The most common interval of relapse is about 2 years. The relapsed disease is usually more widespread and located deeper. Most recurrent NPC is advanced disease.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Antigens, Viral , Blood , Bone Neoplasms , Capsid Proteins , Blood , Immunoglobulin A , Blood , Lung Neoplasms , Lymphatic Metastasis , Nasopharyngeal Neoplasms , Blood , Pathology , Virology , Neoplasm Invasiveness , Neoplasm Recurrence, Local , Pathology , Neoplasm Staging , Retrospective Studies
11.
Chinese Journal of Hematology ; (12): 363-367, 2009.
Article in Chinese | WPRIM | ID: wpr-314480

ABSTRACT

<p><b>OBJECTIVE</b>To study the resistant related molecules of human leukemia drug resistant K562 cells (K562/HHT) induced by homoharringtonine (HHT).</p><p><b>METHODS</b>Gene expression profiles on K562/HHT, K562 and K562/HHT/RU486 (K562/HHT reversed by RU486) cells were detected by DNA microarray. The bone marrow tyrosine kinase gene in chromosome X (BMX) which changed dynamically among the three cells was confirmed by RT-PCR and Western blot. Then, BMX was transfected into K562 and K562/HHT cells, and the changes of daunorubicin (DNR) concentrations in these two cells were observed for BMX overexpression.</p><p><b>RESULTS</b>As compared with K562, there were changes in 117 gene expressions in K562/HHT, 57 of which were up-regulated and 60 down-regulated. The mdrl gene was significantly up-regulated. When compared with K562/HHT, 50 significantly differently expressed genes were screened out in the K562/HHT/RU486 cells, of which up- and down-regulated genes were 13 and 37 respectively. These genes involved in drug resistance, cell signaling, cell differentiation, cell proliferation, transcription regulator, ion transport and so on. Four genes [NM-001721 (BMX), NM-031459 (SESN2), NM-033642 (FGF13) and AL-049309 (SFRS12)] expressed significantly differently in the two group cells, BMX gene expression was higher in K562/HHT, than in K562, but lower than in K562/HHT/RU486 as confirmed by RT-PCR and Western blot. After the plasmid pCI-neo-BMX was transfected into K562 and K562/HHT cells, DNR concentration was significantly lower (79.28 +/- 4.04, 29.84 +/- 2.67) than those before transfection (158.52 +/- 8.08, 58.58 +/- 6.53).</p><p><b>CONCLUSION</b>BMX is associated with multi-drug resistance of K562/HHT cell line.</p>


Subject(s)
Humans , Drug Resistance, Multiple , Genetics , Drug Resistance, Neoplasm , Genetics , Gene Expression Profiling , Harringtonines , Pharmacology , K562 Cells , Leukemia , Drug Therapy , Genetics , Metabolism , Protein-Tyrosine Kinases , Genetics , Metabolism
12.
Microbiology ; (12)2008.
Article in Chinese | WPRIM | ID: wpr-686023

ABSTRACT

PBL is a modern model of classroom teaching. We have introduced it into the teaching of Microbiology. As a result, the students’ learning abilities have been raised to a higher level, and their learning autonomy and achievement have been improved. The combination of PBL method with traditional teaching methods achieved a good effect.

13.
Chinese Journal of Hematology ; (12): 555-559, 2007.
Article in Chinese | WPRIM | ID: wpr-262984

ABSTRACT

<p><b>OBJECTIVE</b>To study whether progestogen antagonist mifepristone could reverse multidrug resistance of K562/A02 cells and its mechanisms.</p><p><b>METHODS</b>MTT was used to study the proliferation of K562/A02 cells and sensitivity of K562/A02 cells to ADM after 72 hours treatment with mifepristone. Flow cytometry was used to assay the expression of P-glycoprotein and the mean fluorescent intensity of intracellular daunorubicin. The expressions of apoptosis related proteins (bcl-2, Bax and caspase-3) were assayed by immunohistochemistry and the glucosylceramide synthase mRNA expression by RT-PCR before and after mifepristone treatment.</p><p><b>RESULTS</b>MTT assay revealed that 2.5, 5.0 and 10 micromol/L mifepristone did not affect the proliferation of K562/A02 cells, but enhanced the sensitivity of K562/A02 cells to ADM, by 1. 68-, 4.17- and 10.71- fold increase, respectively. Expression of P-gp in K562/A02 cells was (49.03 +/- 5.32)%, and was decreased to (28.60 +/- 2.13)% (P < 0.01) after 10 micromol/L mifepristone treatment for 72 hours. and intracellular DNR accumulation in K562/A02 was (61.07 +/- 8.61)%, and was increased to (92.72 +/- 3.48)% (P < 0.01). After 10 micromol/L mifepristone treatment, the expression of bcl-2 protein was decreased from (56 +/- 9)% to (37 +/- 6)% (P < 0.05), Bax and caspase-3 proteins was increased from (40 +/- 5)% to (87 +/- 10)% (P < 0.01), and from (36 +/- 7)% to (89 +/- 6)% (P < 0.01) respectively. RT-PCR analysis revealed that expression of glucosylceramide synthase mRNA was higher in K562/A02 than in K562 cells, whereas 10 micromol/L mifepristone significantly down-regulated its expression in K562/A02 cells.</p><p><b>CONCLUSION</b>Mifepristone at 10 micromol/L could dose-dependently reverse the multidrug resistance of K562/A02 cells. The possible mechanisms are related with decreasing the expression of P-gp, regulating the expression of apoptosis related proteins and decreasing the expression of glucosylceramide synthase.</p>


Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Metabolism , Cell Proliferation , Daunorubicin , Pharmacokinetics , Doxorubicin , Pharmacology , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Glucosyltransferases , Genetics , Metabolism , K562 Cells , Mifepristone , Pharmacology , RNA, Messenger , Genetics , bcl-2-Associated X Protein , Metabolism
14.
Chinese Journal of Hematology ; (12): 818-822, 2007.
Article in Chinese | WPRIM | ID: wpr-262943

ABSTRACT

<p><b>OBJECTIVE</b>To study the maturation effect of CpG2006 and phosphodiester oligonucleotides on leukemia-derived dendritic cells.</p><p><b>METHODS</b>Leukemia cells K562/A02 were induced into dendritic cells by rhGM-CSF and rhIL-4. After 7 days induction, the cell-morphology was observed, the immunophenotype of cells was detected by flow cytometry and the cell function was evaluated by allogeneic mixed lymphocyte reactions, CTL responses and secretion of IL-12 and IL-6. Then a CpG oligonucleotide CpG2006, two synthetic bacterial phosphodiester oligonucleotides A-ODN and T-ODN were added to these leukemia-derived DCs. Three days later, the DCs were re-detected by the above-mentioned methods.</p><p><b>RESULTS</b>After induced by CpG2006, A-ODN or T-ODN, the leukemia-derived DCs with typical dendritic morphology were increased. The expressions of CD83, HLA-DR and CD86 were (65.5 +/- 8.4)%, (32.0 +/- 4.3)% and (18.6 +/- 3.2)% respectively in day 7 leukemia-derived DCs, raised to (88.9 +/- 3.6)%, (53.9 +/- 3.2)% and (39.9 +/- 7.3)% respectively after exposing CpG2006 for 3 days; increased to (97.0 +/- 5.3)%, (63.9 +/- 7.3)% and (40.2 +/- 7.4)% respectively after treated by A-ODN; and further increased to (93.26 +/- 4.65)%, (58.3 +/- 5.6)% and (36.2 +/- 6.8)% respectively after treated by T-ODN. These results was markedly different than unaffected cells did. These DCs induced by the above-mentioned three oligonucleotides could upregulate significantly the capacity for stimulating allogeneic T cells. They could also induce CTL to generate specific cytotoxic activity against K562/A02 cells. The secretion of IL-6 and IL-12 was increased remarkably.</p><p><b>CONCLUSION</b>CpG2006, as well as two phosphodiester oligonucleotides can induce leukemia-derived DCs maturation.</p>


Subject(s)
Humans , Cell Differentiation , Cell Survival , Dendritic Cells , Cell Biology , K562 Cells , Oligodeoxyribonucleotides , Pharmacology , Oligonucleotides , Pharmacology
15.
Chinese Journal of Medical Genetics ; (6): 238-239, 2006.
Article in Chinese | WPRIM | ID: wpr-263806

ABSTRACT

<p><b>OBJECTIVE</b>To research and compare HLA-DQB1 gene frequency(GF) and polymorphism distribution between south and north population of Chinese in China.</p><p><b>METHODS</b>Combining PCR-sequence specific primers(SSP) and sequence specific oligonucleotide probe(SSOP) techniques and DNA Microarray Kit for HLA-DQB1 Low Res Genotyping from Shenzhen Yi-Shengtang Biological LTD. Co. was used to type HLA-DQB1 gene polymorphisms of 700 individuals living in south China and 320 individuals in north China.</p><p><b>RESULTS</b>We inspected 10 alleles of HLA-DQB1 and got a series of comprehensive and accurate statistic data.</p><p><b>CONCLUSION</b>It is tested that HLA-DQB1*02, 05, 0601, 0602, 0603 gene frequencies are different obviously(P<0.05) between south and north Chinese. And those data will be useful to kinds of research associated with disease relevant and anthropology research.</p>


Subject(s)
Female , Humans , Male , Alleles , Asian People , Genetics , China , Ethnology , Gene Frequency , Genetic Variation , HLA-DQ Antigens , Genetics , HLA-DQ beta-Chains , Oligonucleotide Array Sequence Analysis , Methods , Oligonucleotide Probes , Polymerase Chain Reaction , Polymorphism, Genetic , Sequence Analysis, DNA
16.
Chinese Medical Journal ; (24): 1151-1154, 2004.
Article in English | WPRIM | ID: wpr-291962

ABSTRACT

<p><b>BACKGROUND</b>Cyclo-oxgenase 2 (COX-2) is involved in prostaglandin synthesis in central nervous system, and it also plays a role in human carcinogenesis. Our purpose of this study is to investigate the COX-2 expression in different development stages of colorectal cancer, and to discuss the relationship between the gene expression and clinicopathological features of the cancer.</p><p><b>METHODS</b>COX-2 expression was examined by immunohistochemical staining in 76 surgical specimens of colorectal cancer (44 of advanced stage and 32 of early stage), thirty-three adenomas and 18 normal colonic mucosal tissues taken by endoscopic biopsy. Kaplan-Meier survival curves and Cox proportional hazards regression were used to evaluate the relation of COX-2 to prognosis.</p><p><b>RESULTS</b>COX-2 expression, divided into 4 grades from "-" to "+++", is respectively 83.3%, 16.7%, 0% and 0% in normal colonic mucosal tissues; 12.1%, 42.4%, 36.4% and 9.1% in adenomas; 6.3%, 28.1%, 46.9% and 18.7% in early colorectal cancers (ECCs), and 6.8%, 20.5%, 18.2% and 54.5% in advanced colorectal cancers (CRCs). The differences in COX-2 expression between advanced CRCs and early colorectal cancers (ECCs) as well as between the advanced CRCs and adenomas were statistically significant (P < 0.01); but there was no significant difference between ECCs and adenomas. Kaplan-Meier survival analysis showed a significant difference in the survival curves between low high COX-2 groups (P < 0.05). Cox proportional hazards regression showed that COX-2 expression was related to poorer long-term outcome with a hazard ratio of 2.665 unadjusted for other variables (P < 0.05), and COX-2 expression was an independent risk factor of poor prognosis.</p><p><b>CONCLUSIONS</b>COX-2 expression is gradually up-regulated in the development from normal epithelium to adenomas and from ECCs to advanced CRCs. Alhough the COX-2 protein can not be regarded as a tumor marker to diagnose CRCs early, COX-2 expression can be regarded as an independent risk factor of poor prognosis for postoperative patients with advanced CRCs.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Colorectal Neoplasms , Mortality , Pathology , Cyclooxygenase 2 , Immunohistochemistry , Isoenzymes , Membrane Proteins , Prognosis , Prostaglandin-Endoperoxide Synthases , Survival Rate
17.
Chinese Journal of Hematology ; (12): 13-16, 2004.
Article in Chinese | WPRIM | ID: wpr-291461

ABSTRACT

<p><b>OBJECTIVE</b>To observe the antileukemic effect of lymphocytes from cord blood treated by CpG-oligodeoxynucleotides (CpG-ODN).</p><p><b>METHODS</b>Lymphocytes from cord blood were exposed to different oligodeoxynucleotides containing a panel of CpG-ODN and were cultured with K562 cells. The cytotoxic effects were detected by MTT method. Immunological markers of cord blood treated by CpG-ODN(3) which showed highest activity were measured with flow cytometry.</p><p><b>RESULTS</b>Different CpG-motifs have different immunostimulatory activity and CpG-ODN(3) has the highest one. After treated by CpG-ODN(3), NK killing activity to K562 cells increased in a dose-dependent manner, and CD(3), CD(4), CD(19) and CD(56) increased to (60.6 +/- 7.9)%, (40.2 +/- 3.5)%, (22.4 +/- 1.9)% and (15.5 +/- 3.1)%, respectively.</p><p><b>CONCLUSION</b>CpG-ODN could reinforces the immunological competence of cord blood lymphocytes and their effects on K562 cells. This provides a new approach to reinforce the antitumor effects of cord blood.</p>


Subject(s)
Humans , Adjuvants, Immunologic , Pharmacology , Antigens, CD , Blood , Cytotoxicity, Immunologic , Dose-Response Relationship, Drug , Fetal Blood , Cell Biology , K562 Cells , Leukemia , Therapeutics , Lymphocytes , Allergy and Immunology , Oligodeoxyribonucleotides , Pharmacology , Reverse Transcriptase Polymerase Chain Reaction
18.
Chinese Journal of Hematology ; (12): 624-628, 2003.
Article in Chinese | WPRIM | ID: wpr-354815

ABSTRACT

<p><b>OBJECTIVE</b>To study the related proteins of apoptosis initiation induced by homoharringtonine (HHT) in HL-60 cells.</p><p><b>METHODS</b>After establishment of an apoptosis initiation model induced by HHT in HL-60 cells, proteins of untreated and HHT treated HL-60 cells were extracted, and the two-dimensional polyacrylamide gel electrophoresis (2-DE) maps of the extracted proteins were established by using the immobilized pH gradient (IPG) two-dimensional electrophoresis respectively. The alteration protein spots were identified with assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database searching.</p><p><b>RESULTS</b>Proteomics analysis showed that proteins including MHC class I antigen, calbindin D-28K, chloride channel protein 6, oncoprotein 18, zinc finger protein Helios and apoptosis inhibitor like protein 2 were involved in apoptosis initiation induced by HHT.</p><p><b>CONCLUSION</b>The present study might conduce to the researches of HL-60 cells carcinogenesis and pave the way to exploit drug precursor related to HHT and initiation of apoptosis in HL-60 cells.</p>


Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Calbindins , Chloride Channels , DNA-Binding Proteins , Electrophoresis, Gel, Two-Dimensional , Methods , HL-60 Cells , Harringtonines , Pharmacology , Histocompatibility Antigens Class I , Ikaros Transcription Factor , Inhibitor of Apoptosis Proteins , Microtubule Proteins , Phosphoproteins , Proteins , Proteome , S100 Calcium Binding Protein G , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Stathmin , Transcription Factors
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