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1.
Chinese Journal of Hematology ; (12): 28-31, 2018.
Article in Chinese | WPRIM | ID: wpr-1011681

ABSTRACT

Objective: To observe the effect of poloxamer 188 (P188) on megakaryocyte cultivation and induction from cord blood mononuclear cells in order to obtain more megakaryocyte progenitor cells (MPC). Methods: The cord blood mononuclear cells were isolated and inoculated in cell culture bag or cell culture flask respectively. The WIGGENS shaker and cell culture bags were used to mimick WAVE Bioreactor for three-dimensional (3D) cell culture, and the P188 was added to induction medium, The cells were detected for morphology, surface marker, viability, and number on day 14. Results: In the two-dimensional (2D) culture, CD41(+), CD41(+)/CD61(+), CD61(+) megakaryocytic numbers increased significantly after adding P188 (all P<0.01). And in the 3D culture of adding P188, the cell volume became larger and the nuclear shape was irregular, the cytoplasm appeared magenta granules, and the megakaryocyte cells became more mature. By 3D culture, the expression of CD41/CD61 was (36.30±1.27)% vs (23.95±1.34)%, hence the differentiation for MPC was significantly higher than that in the 2D group (P<0.01). Furthermore, adding P188 in 3D culture resulted in highest differentiation efficiency for MPC [(59.45±1.20)%]. There were no significantly differences in terms of cell viability and cell number among 3D culture containing P188, 2D and 3D culture groups (all P>0.05). Conclusion: 3D culture was beneficial for the differentiation of MPC, but the cell viability was lower than 2D group; However, the satisfied cell growth and better induction efficiency were obtained by adding of P188, which might provide a new method of megakaryocytes production for clinical application.


Subject(s)
Bioreactors , Cell Differentiation , Cells, Cultured , Fetal Blood , Megakaryocytes , Poloxamer
2.
Chinese Journal of Virology ; (6): 148-153, 2014.
Article in Chinese | WPRIM | ID: wpr-356623

ABSTRACT

The JC virus is a widely infected human polyomavirus. Recent foreign researches showed that the JC virus infection is correlated with tumors of nervous system and digestive system, while, and study on the relationship between JC virus infection and gynecological tumor is seldom reported. In this study, we first establish the nucleic acid detection methods and procedures for JC virus and its highly homologous BK virus. The JC and BK viruses infection was evaluated by detect the viral DNA in samples including biopsy tissues, serum as well as urine of myoma of uterus (98 cases), cervical cancer (84 cases), endometrial cancer (40 cases) and ovarian tumor (72 cases) patients. The BK viral DNA positive rate was significantly higher in urine samples than that of blood and biopsy samples, and there is no significant difference of the BK viral DNA positive rate among all patient groups. The JC viral DNA positive rate is almost 0 in serum samples and biopsy. tissues, however, viral DNA positive rate is more than 50% in urine samples. In fibroids group, the JC viral DNA positive rate is up to 65. 3% which is significantly higher than that in other patients groups and healthy control. Further gynecological tumor associated viruses detection showed that only human papilloma virus infection is associated with cervical cancer, the herpes simplex virus, EB virus and cytomegalovirus infection is extremely low in our patient groups. No synergistic effect on gynecological tumor caused by viruses co-infection was observed. Our study showed that JC virus infection is highly related to the pathogenesis of uterine fibroids.


Subject(s)
Adult , Female , Humans , Middle Aged , Young Adult , Genital Neoplasms, Female , Virology , JC Virus , Classification , Genetics , Polyomavirus Infections , Virology , Tumor Virus Infections , Virology
3.
Acta Academiae Medicinae Sinicae ; (6): 402-407, 2011.
Article in Chinese | WPRIM | ID: wpr-341394

ABSTRACT

<p><b>OBJECTIVE</b>To explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells (BMSCs) in vivo.</p><p><b>METHODS</b>Melanocytes were isolated from human foreskin. BMSCs were isolated from human bone marrow. Both of them were co-cultured at a ratio of 1:10, and then were implanted into the collagen membrane to construct the tissue-engineered skin, which was applied for wound repair in nude mice. The effectiveness of wound repair and the distribution of melanocytes were evaluated by morphological observation, in vivo 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.</p><p><b>RESULTS</b>The wounds were satisfactorily repaired among the nude mice. The melanocytes were distributed in the skin with normal structure, as confirmed by DAPI fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.</p><p><b>CONCLUSION</b>Melanocytes and BMSCs, after proper in vitro culture at an appropriate ratio, can construct the tissue-engineered skin with I type collagen membrane.</p>


Subject(s)
Animals , Humans , Mice , Bone Marrow Cells , Cell Biology , Cells, Cultured , Coculture Techniques , Collagen Type I , Melanocytes , Cell Biology , Mesenchymal Stem Cells , Cell Biology , Mice, Nude , Skin , Wounds and Injuries , Skin, Artificial , Tissue Engineering
4.
Acta Academiae Medicinae Sinicae ; (6): 659-661, 2005.
Article in Chinese | WPRIM | ID: wpr-318843

ABSTRACT

Cancer stem cells are defined as rare cells in cancer tissues with indefinite potential for self-renewal that drives tumorigenesis. It was first extensively documented for leukaemia and multiple myeloma. It has also been found in solid cancers such as human breast cancer and nervous system tumors. Studies of cancer stem cell biology and mechanisms of tumorigenesis are lending insight into the origins of cancer and will ultimately yield new approaches to fight cancer.


Subject(s)
Animals , Humans , Mice , Cell Transformation, Neoplastic , Neoplasms , Pathology , Therapeutics , Neoplastic Stem Cells , Cell Biology , Tumor Stem Cell Assay , Methods
5.
Chinese Journal of Stomatology ; (12): 418-420, 2004.
Article in Chinese | WPRIM | ID: wpr-324131

ABSTRACT

<p><b>OBJECTIVE</b>PPTA and c-fos mRNA expression were detected in dog caudalis subnucleus of trigeminal spinal tract nucleus (VC) induced by trauma occlusion in order to investigate orofacial pain mechanism.</p><p><b>METHODS</b>The occlusal surface of the first and second maxillary right molars in 15 dogs were unilaterally raised 1.5 mm with casting Ni-Cr inlay which were fixed in Class I hole. On days 3, 7, 14, 30 and 60 after teeth operation, the VC of right and left sides were removed. PPTA and c-fos mRNAs were detected in experimental and control groups with reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) The basal levels of PPTA and c-fos mRNAs were extremely low and poorly detectable in VC in control animals. (2) The expression of PPTA mRNA in VC of traumatic side was up regulated from 3 days after inlay was fixed in molar and reached peak level during 14 to 30 days and then down-regulated gradually and no significant difference was noted between 60 days group and control group. (3) c-fos mRNA expression was more intense during 3 to 7 days compared with the control group but undetectable in the other experimental period. (4) Both PPTA and c-fos mRNAs expression in VC of trauma occlusal side were more intense than that in the contralateral side.</p><p><b>CONCLUSIONS</b>The present results show that both PPTA and c-fos mRNA expression are elevated in dog's VC induced by traumatic occlusion. The primary afferent terminal of orofacial area is sensitized, which suggest one kind of mechanism of orofacial pain in the condition of traumatic occlusion.</p>


Subject(s)
Animals , Dogs , Dental Occlusion, Traumatic , Facial Neuralgia , Protein Precursors , Genetics , Proto-Oncogene Proteins c-fos , Genetics , RNA, Messenger , Tachykinins , Genetics , Trigeminal Nucleus, Spinal , Metabolism
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