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1.
Chinese Acupuncture & Moxibustion ; (12): 857-862, 2020.
Article in Chinese | WPRIM | ID: wpr-826642

ABSTRACT

OBJECTIVE@#To observe the effect of moxibustion on oxidative stress injury of nigrostriatal system in rats with Parkinson's disease (PD) based on nuclear factor erythroid 2-related factor (Nrf2)/antioxidant response element (ARE) pathway, and to explore its mechanism.@*METHODS@#A total of 48 SD rats were randomized into a blank group, a sham-operation group, a model group and a moxibustion group, 12 rats in each group. Unilateral two-point injection with 6-hydroxydopamine (6-OHDA) was adopted in the model group and the moxibustion group to establish the PD model; the operation manipulation in the sham-operation group was the same as the model group and the moxibustion group, and the same volume of 0.9% sodium chloride solutions was given by unilateral two-point injection. Moxibustion was adopted at "Baihui" (GV 20) and "Sishencong" (EX-HN 1) in the moxibustion group for 20 min, once a day, 6 times a week for 6 weeks. No intervention was given in the other 3 groups. Morphology of right mesencephalon substantia nigra was observed by HE staining, the expression of tyrosine hydroxylase (TH) in right mesencephalon substantia nigra was detected by immunohistochemistry method, the expression of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-Px) in corpus striatum was detected by colorimetry method, and the expression of Nrf2 and heme oxygenase-1 (HO-1) proteins was detected by Western blot in the 4 groups.@*RESULTS@#Clear tissue structure and complete dopaminergic neurons of right mesencephalon substantia nigra were observed in the blank group and the sham-operation group; unclear tissue structure, decreased and swelling dopaminergic neurons were observed in the model group; compared with the model group, more neurons were observed and the swelling of cyton was reduced in the moxibustion group. Compared with the sham-operation group, the expression of TH in right mesencephalon substantia nigra was decreased in the model group (<0.01); compared with the model group, the expression of TH in right mesencephalon substantia nigra was increased in the moxibustion group (<0.05). Compared with the sham-operation group, the expression of ROS, MDA was increased (<0.01), the expression of GSH, GSH-Px, Nrf2 and HO-1 was decreased in the model group (<0.01, <0.05); compared with the model group, the expression of ROS, MDA was decreased (<0.05, <0.01), the expression of GSH, GSH-Px, Nrf2 and HO-1 was increased in the moxibustion group (<0.05, <0.01).@*CONCLUSION@#Moxibustion can alleviate oxidative stress injury of nigrostriatal system in rats with Parkinson's disease by activating the Nrf2/ARE pathway, and protect the dopamine neurons.

2.
Chinese Journal of Information on Traditional Chinese Medicine ; (12): 104-109, 2018.
Article in Chinese | WPRIM | ID: wpr-707002

ABSTRACT

Objective To conduct Meta analysis and trial sequential analysis (TSA) on warm acupuncture treatment on lumbar intervertebral disc protrusion (LIDP); To provide references for evidence-based medicine of this disease.Methods Articles about warm acupuncture treatment for LIDP clinical randomized controlled trials in CNKI, Wanfang database, Chonging Wepu, CBM, OubMed, Cochrane Library, and Emnase were retrieved by computer. The retrieval range was from the database establishing to March, 2017. According to Cochrane Handbook for Systematic Reviews of Interventions 5.2.0 Bias risk assessment tool, included articles were under quality evaluation. Revman5.2 software was used to carry out Meta analysis, and TSAv0.9 software was used to conduct TSA.Results Ten articles were included, involving 1035 cases. Meta analysis showed that the total effective rates of warm acupuncture treatment for LIDP [95%CI (2.43, 5.40),Z=6.31,P<0.00001], pain index [95%CI (-1.05, -0.58),Z=6.77,P<0.00001], lumbar function [95%CI (2.56, 8.61),Z=3.62,P=0.0003] were better than other therapies, with statistical significance. Funnels included in the study suggest publication bias. TSA results suggested that the total efficiency and pain index Meta analysis results of this study were reliable.Conclusion Warm acupuncture treatment for LIDP has confirmed efficacy, with certain advantages. However, the literature included is not with good quality, so more large sample, multicenter, methodological RCTs are needed for further validation.

3.
Chinese Journal of Clinical and Experimental Pathology ; (12): 137-142, 2018.
Article in Chinese | WPRIM | ID: wpr-695071

ABSTRACT

Purpose To investigate the histopathological diagnosis and follow-up of gastric mucosal biopsy. Methods A detailed histopathological observation was performed on 5 748 gastroscopic biopsies and 3 288 patients were followed up. Results In 5 748 cases of endoscopy biopsy specimens, acute gastritis was rechecked in 125 cases, of which 112 cases were cured or improved (89.6% ), 10 cases (8.0% ) had little change, and 3 cases (2.4%) were aggravated. There were 2430 cases of chronic atrophic gastritis, in which 1521 cases were cured/improvement (62.6% ), 737 cases (30.3% ) had little change, and 172 cases (7.1% ) were aggravated. In the 71 cases of special type of gastritis, 23 cases were cured or improved, accounting for 32.3%.42 cases were not changed much, accounting for 59.2%, and 6 cases were aggravated (8.5% ). About druginduced gastritis/stomach reexamination in 94 cases, 57 cases were cured/improved, accounting for 60.6%, 34(36.3% ) cases did not change, and 2 cases were aggravated(3.2% ).183 cases of gastric polyp were reviewed, 165 cases(90.2% ) were cured / improved, 13 cases (7.1% ) were not changed and 5 cases (2.7% ) were aggravated. No neoplasia/ nondysplasia was found in 205 cases, 196(95.6% ) cases was cured / improved, no change was seen in 6 cases (2.9% ) and aggravation was seen in 3 cases(1.5%). Indefinite neoplasia/ dysplasia was reexamined in 24 cases, 2 cases (8.3%) were cured/improved, no change was seen in 13 cases (54.2% ), and aggravation was seen in 9 cases(37.5%).156 cases of intraepithelial neoplasia were retrospectively reviewed, in which138 (88.5% ) cases were cured or improved, 4(2.6% ) cases showed no change, and 13(8.3%) cases were aggravated. Conclusion It is of great significance to improve the early diagnosis rate, reduce missed diagnosis rate and misdiagnosis rate of gastric cancer by establishing 11 pathological changes and accompanying lesions.

4.
China Journal of Chinese Materia Medica ; (24): 1545-1550, 2017.
Article in Chinese | WPRIM | ID: wpr-350150

ABSTRACT

One of the top-level researches of biopharmaceutics classification system of Chinese materia medica (CMMBCS) is the study on single component in compound Chinese medicine. The medicines shall be classified according to its solubility and intestinal permeability, as well as the ascending degree in multicomponent environment. Based on above, we chose berberine as the main object to explore the change rules of its solubility and intestinal permeability in Gegen Qinlian decoction. Shaking flask-HPLC was used to detect the solubility changes of berberine in compounds. The qualitative investigation of berberine in intestinal absorption was measured by everted gut sac, and the quantitative research of berberine in intestinal absorption was measured by single-pass intestinal perfusion experiment, while the qualitative and quantitative research of berberine absorption into blood was measured by in intestinal perfusion with venous sampling experiment.

5.
China Journal of Chinese Materia Medica ; (24): 1192-1197, 2016.
Article in Chinese | WPRIM | ID: wpr-320879

ABSTRACT

The study of single component in the multicomponent environment is one of the basic researches for biopharmaceutics classification system of Chinese materia medica (CMMBCS). That is to say, the classification research shall be based on the respective lift of solubility and permeability in the multicomponent environment, besides solubility and intestinal permeability of the single component. We chose berberine as the main research object to investigate the changes of its solubility and intestinal permeability in Huanglian decoction. Shake-flask and HPLC were used to detect the solubility of berberine in different pH buffer solutions and different concentrations of Huanglian decoction. In situ single-pass intestinal perfusion (SPIP) and intestinal perfusion with venous sampling (IPVS) were carried out to study berberine's intestinal absorption and absorption into blood, respectively.

6.
Chinese Journal of Cancer ; (12): 573-582, 2015.
Article in English | WPRIM | ID: wpr-349558

ABSTRACT

<p><b>BACKGROUND</b>Serine hydroxymethyltransferase 1 (SHMT1) is a key enzyme in the folate metabolic pathway that plays an important role in biosynthesis by providing one carbon unit. SHMT1 C1420T may lead to the abnormal biosynthesis involved in DNA synthesis and methylation, and it may eventually increase cancer susceptibility. Many epidemiologic studies have explored the association between C1420T polymorphism and the risk of non-Hodgkin lymphoma (NHL), but the results have been contradictory. Therefore, we performed this meta-analysis to evaluate the relationship.</p><p><b>METHODS</b>The meta-analyses were conducted to evaluate the effect of SHMT1 C1420T polymorphism on NHL risk. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to measure the strength of the association.</p><p><b>RESULTS</b>Eight studies encompassing 3232 cases and 4077 controls were included. A statistically significant association was found between SHMT1 C1420T polymorphism and NHL risk under the allelic comparison (T vs. C: OR = 1.09, 95% CI 1.01-1.17); a borderline association was found between SHMT1 C1420T polymorphism and NHL risk under the homozygote model (TT vs. CC: OR = 1.18, 95% CI 1.00-1.39) and the dominant model (CT+TT vs. CC: OR = 1.10, 95% CI 1.00-1.21).</p><p><b>CONCLUSION</b>SHMT1 C1420T polymorphism may be associated with NHL risk, which needs to be validated in large, prospective studies.</p>


Subject(s)
Humans , Case-Control Studies , Evidence-Based Medicine , Methods , Genetic Predisposition to Disease , Glycine Hydroxymethyltransferase , Genetics , Lymphoma, Non-Hodgkin , Genetics , Neoplasm Proteins , Genetics , Polymorphism, Single Nucleotide , Publication Bias , Sensitivity and Specificity
7.
China Journal of Chinese Materia Medica ; (24): 4479-4483, 2014.
Article in Chinese | WPRIM | ID: wpr-341794

ABSTRACT

Based on the characteristics of multicomponent of traditional Chinese medicine and drawing lessons from the concepts, methods and techniques of biopharmaceutics classification system (BCS) in chemical field, this study comes up with the science framework of biopharmaceutics classification system of Chinese materia medica (CMMBCS). Using the different comparison method of multicomponent level and the CMMBCS method of overall traditional Chinese medicine, the study constructs the method process while setting forth academic thoughts and analyzing theory. The basic role of this system is clear to reveal the interaction and the related absorption mechanism of multicomponent in traditional Chinese medicine. It also provides new ideas and methods for improving the quality of Chinese materia medica and the development of new drug research.


Subject(s)
Animals , Humans , Biopharmaceutics , China , Drugs, Chinese Herbal , Chemistry , Classification , Pharmacology , Materia Medica , Chemistry , Classification , Plants, Medicinal , Chemistry , Classification
8.
China Journal of Chinese Materia Medica ; (24): 4489-4493, 2014.
Article in Chinese | WPRIM | ID: wpr-341792

ABSTRACT

The complex level of constructing biopharmaceutics classification system of Chinese materia medica CMMBCS) was the study of traditional Chinese compound, on the premise of insisting that the multicomponent simultaneous determination, when carrying out the study of intestinal permeability, the primary task was to define the source of the components that was absorbed through the intestinal wall, namely, which medicinal material the components belonged to in traditional Chinese compound. The technology of chemical fingerprint and in vitro everted gut sac model were used in this research to make multicomponent an intuitive source attribution which permeated the intestine in the classic formula Gegen Qinlian decoction, and to lay the foundation for the further qualitative and quantitative research of intestinal permeability.


Subject(s)
Animals , Male , Rats , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Pharmacokinetics , Intestines , Metabolism , Permeability , Plants, Medicinal , Chemistry , Rats, Wistar
9.
China Journal of Chinese Materia Medica ; (24): 4494-4498, 2014.
Article in Chinese | WPRIM | ID: wpr-341791

ABSTRACT

The study is a paticular embodiment of Chinese patent medicine based on biopharmaceutics classification system of Chinese materia medica (CMMBCS) , focusing on assessment of synchronization issues of dissolution that may affect the timing of the multicomponent absorption. The accumulative dissolution percentages of nine components in Gengen Qinlian tablets in different dissolution solvents and times were determined by HPLC. The dissolution curve was drew and its similarity was evaluated by similarity factors (f2) and cluster method. Results in this experiment showed that the components that peak 7 and peak 8 (baicalin) represented had poor similarity with the reference peak 2 (puerarin). Their similarity factors were both 43 in water dissolution media and 31 and 45 in pH 7.4 dissolution media, respectively. Components that peaks represented had better similarity with the reference peak 2 (puerarin) in other medium. It illustrated that components that peak 3,4,5,6 (berberine) represented had fully synchronous dissolution characteristics with the reference peak 2 (puerarin), components peak 1 and 9 represented had nearly fully synchronous dissolution characteristics with the reference peak 2 (puerarin), while components that peak 7 and 8 (baicalin) represented had no synchronous dissolution characteristics with the reference peak 2 (puerarin).


Subject(s)
Biopharmaceutics , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Classification , Hydrogen-Ion Concentration , Solubility , Tablets , Chemistry , Classification
10.
China Journal of Chinese Materia Medica ; (24): 4505-4508, 2014.
Article in Chinese | WPRIM | ID: wpr-341789

ABSTRACT

The evaluation of permeability in biopharmaceutics classification system of Chinese materia medica (CMMBCS) requires multicomponent as a whole in order to conduct research, even in the study of a specific component, should also be put in the multicomponent environment. Based on this principle, the high content components in Gegen Qinlian decoction were used as multicomponent environmental impact factors in the experiment, and the relevant parameters of intestinal permeability about puerarin were measured with using in situ single-pass intestinal perfusion model, to investigate and evaluate the intestinal permeability of puerarin with other high content components. The experimental results showed that different proportions of baicalin, glycyrrhizic acid and berberine had certain influence on intestinal permeability of puerarin, and glycyrrhizic acid could significantly inhibit the intestinal absorption of puerarin, moreover, high concentration of berberine could promote the absorption of puerarin. The research results indicated that the important research ideas of permeability evaluation in biopharmaceutics classification system of Chinese materia medica with fully considering the effects of other ingredients in multicomponent environment.


Subject(s)
Animals , Male , Rats , Berberine , Pharmacokinetics , Biopharmaceutics , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacokinetics , Flavonoids , Pharmacokinetics , Glycyrrhizic Acid , Pharmacokinetics , Intestines , Chemistry , Metabolism , Isoflavones , Pharmacokinetics , Kinetics , Materia Medica , Pharmacokinetics , Permeability , Rats, Wistar
11.
Chinese Journal of Medical Genetics ; (6): 694-697, 2005.
Article in Chinese | WPRIM | ID: wpr-279965

ABSTRACT

<p><b>OBJECTIVE</b>It is demonstrated that KRAS2, functioning as an oncogene, plays a critical role in carcinogenesis. However, some studies suggest that the wild type KRAS2, located in the region of 12p12.1, takes its effect as a tumor suppressor gene. This study, therefore, is aimed to investigate the loss of heterozygosity (LOH) on chromosome 12p12-13 region in 10 human colon carcinomas.</p><p><b>METHODS</b>LOH analysis of the 12p12-13 region was performed by PCR, using 11 microsatellite markers in 12p12-13 region. The relationships between LOH for each marker and clinical pathologic factors were evaluated.</p><p><b>RESULTS</b>LOH in at least one of the loci in 12p12-13 region was detected in 30% (3/10) of adjacent tissues; the highest frequency of LOH was identified at the locus of D12S1034 in 28.57% (2/7) of adjacent tissues. 60% (6/10) carcinoma tissues were found to have LOH in at least one locus in the same region; the most frequent LOH was found at the loci of D12S1034 and D12S1591, both about 42.86% (3/7). Among all samples, 3 cases were noted to have LOH in both adjacent and tumor tissues, and 3 cases were shown to have LOH only in tumor tissues. Occurrence of LOH was not correlated with sex, age, tumor size and lymph node metastasis.</p><p><b>CONCLUSION</b>Allelic loss on 12p12-13 region would influence the KRAS2 expression by reducing the gene-dosage in colon carcinogenesis.</p>


Subject(s)
Humans , Asian People , Genetics , China , Chromosomes, Human, Pair 12 , Genetics , Colonic Neoplasms , Ethnology , Genetics , Loss of Heterozygosity , Polymerase Chain Reaction , Proto-Oncogene Proteins , Genetics , Proto-Oncogene Proteins p21(ras) , ras Proteins , Genetics
12.
Journal of Experimental Hematology ; (6): 397-403, 2005.
Article in Chinese | WPRIM | ID: wpr-356550

ABSTRACT

To investigate effects of rat bone marrow mesenchymal stem cells (rBMMSC) on hematopoiesis after allo-hematopoietic stem cell transplantation (HSCT), allogeneic BMT model from Fischer 344 rats (RT-1Al) to Wistar rats (RT-1Au) was established; effects of MSCs on hematopoietic reconstitution were studied by survival rate, peripheral blood counts, histological analysis and FACS at day 30 after transplantation. The results showed that (1) MSCs from donor Fisher344 could survive in recipient irradiated by lethal dose and could be found in the thymus, spleen and bone marrow of the recipient at 30 days after cotransplantation with BM by measuring EGFP gene. (2) Cotransplanation of MSCs and BM improved hematopoietic reconstitution. Lymphocyte and platelet counts of peripheral blood in cotransplantation group were higher than those in the control group. Active hematopoiesis and increase of bone marrow nucleated cells were observed in cotransplantation group. MSCs significantly enhanced hematopoiesis of B lymphocyte and megakaryocytopoietic lineages by FACS analysis. It is concluded that (1) MSCs of Fisher344 can be found in the thymus, spleen, bone marrow of the recipients at 30 days after cotransplantion by measuring EGFP gene. (2) hematopoietic reconstitution is significantly enhanced by MSCs cotransplanted with BM.


Subject(s)
Animals , Male , Rats , Bone Marrow Transplantation , Methods , Cell Differentiation , Physiology , Flow Cytometry , Hematopoiesis , Physiology , Lymphocyte Count , Mesenchymal Stem Cell Transplantation , Methods , Mesenchymal Stem Cells , Cell Biology , Physiology , Models, Animal , Platelet Count , Rats, Inbred F344 , Rats, Wistar
13.
Chinese Journal of Stomatology ; (12): 421-424, 2004.
Article in Chinese | WPRIM | ID: wpr-324130

ABSTRACT

<p><b>OBJECTIVE</b>To investigate changes of two sodium channels, PN(3) and NaN, during orofacial pain by occlusal trauma in rat.</p><p><b>METHODS</b>Expressions of PN(3) mRNA and NaN mRNA in trigeminal ganglion were tested during various periods of persistent occlusal trauma with reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>In groups, including control, occlusal trauma groups, PN(3) mRNA and NaN mRNA were all expressed in trigeminal ganglion neurons. In the control group, there were similar density values bilaterally. In the occlusal trauma group, the density values in gel electrophoresis of PN(3) mRNA and NaN mRNA on the intervention side were slightly greater than those on the control side.</p><p><b>CONCLUSIONS</b>The stimulation of occlusal trauma upregulates expressions of PN(3) mRNA and NaN mRNA, which suggests the signal occurring and conduction of chronic pain by occlusal trauma have the same molecular mechanism of sodium channel as inflammatory pain.</p>


Subject(s)
Animals , Male , Rats , Dental Occlusion, Traumatic , Facial Pain , RNA, Messenger , Rats, Sprague-Dawley , Sodium Channels , Genetics , Trigeminal Ganglion , Metabolism
14.
Chinese Journal of Stomatology ; (12): 432-434, 2003.
Article in Chinese | WPRIM | ID: wpr-253680

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of traumatic occlusion on CGRP-immunoreactive (CGRP-IR) nerve fibres in rat molar pulp and observe the recovery of CGRP-IR nerve fibres after removal of traumatic occlusion.</p><p><b>METHODS</b>To observe immunohistochemically the change of CGRP-IR nerve fibres in molar pulp during traumatic occlusion and after removal.</p><p><b>RESULTS</b>The increase of number, density and morphology of CGRP-IR nerve fibres in traumatic occlusion group was more than in control group, however, the changes of CGRP-IR nerve fibres in removal of traumatic occlusion group were less than in control group.</p><p><b>CONCLUSIONS</b>The changes of CGRP-IR nerve fibres in number, morphology, and density are induced by traumatic occlusion in rat molar pulp, however, the nerve fibres recover to normal by removal of traumatic occlusion.</p>


Subject(s)
Animals , Male , Rats , Calcitonin Gene-Related Peptide , Dental Occlusion, Traumatic , Metabolism , Pathology , Therapeutics , Dental Pulp , Immunohistochemistry , Molar , Nerve Fibers , Chemistry , Rats, Sprague-Dawley
15.
Chinese Journal of Applied Physiology ; (6): 183-185, 2002.
Article in Chinese | WPRIM | ID: wpr-319340

ABSTRACT

<p><b>AIM</b>To study the effects of cryopreservation length on the proliferative potential of hematopoietic cells derived from cord blood.</p><p><b>METHODS</b>Using Dextran-40 and 10% DMSO as cryoprotectants, separated nuclear cells were stored in liquid nitrogen after they were freezed according programme. One month or 4 months later, they were thawed and expanded in serum-free medium for culture and expansion of hematopoietic cell (SFEM) for 5 weeks. Dynamic results were detected every week.</p><p><b>RESULTS</b>At the 5th week of expanding, TNC were expanded for 1499.0 +/- 115.6-folds and 1513.0 +/- 110.4-folds, respectively. CD34+ cells and CFCs reached to their highest level at the 2nd week and at the 3rd week. CD34+ cells were expanded for 63.8 +/- 6.1-folds and 62.4 +/- 5.7-folds, respectively. CFCs were expanded for 53.8 +/- 6.3-folds and 54.8 +/- 6.7-folds, respectively. Between the two kinds of cells, statistical significant difference in proliferative potential wasn't detected.</p><p><b>CONCLUSION</b>In ideal cryopreservative condition, the cryopreservation length would do not affect the proliferative potential of cord blood hematopoietic cells.</p>


Subject(s)
Humans , Cell Proliferation , Cell Survival , Cells, Cultured , Cryopreservation , Methods , Fetal Blood , Cell Biology , Hematopoietic Stem Cells , Cell Biology , Time Factors
16.
Journal of Experimental Hematology ; (6): 355-358, 2002.
Article in Chinese | WPRIM | ID: wpr-337622

ABSTRACT

To study the pathogens incidences in cord blood and the efficiency of different detective methods, 60 samples were drawn and reserved from collected and processed cord blood, respectively. The BACTEC 9050 system, improved Martin/thioglycollate broth (22 degrees C) and thioglycollate broth (35 degrees C) were employed to detected bacteria (including fungus) at the same time. Two hundred and six cord blood serum samples were used to detect the HBV DNA and HCV RNA by molecular biology technique, HBsAg, Anti-HBC, Anti-HCV, Anti-HCMV-IgM, HTLV-1, HTLV-2, HIV-1 and HIV-2 by ELISA and RBC agglutination test were used to detect the TPHA. Results showed that using BACTEC 9050 system, the incidence of bacteria and fungus was 3.33% and 0% respectively in collected cord blood; in processed cord blood, the rates increased to 6.67% and 1.67%, respectively. The sensitivity of BACTEC 9050 was higher than that of Martin/thioglycollate broth (22 degrees C/35 degrees C) culture. In 206 serum samples, the positive rate of HBV DNA was 5.8%, HCV RNA was 2.4%, HBsAg was 2.4%, HCMV-IgM was 1.89%, HCV was 2.4% and Anti-HBC was 29.4%. In those samples that Anti-HBC was positive, the positive rate of HBV DNA was 6.7%. It was concluded that the incidences of microbiological contamination in cord blood were high. The routine culture system would lead to false negative results of obligate anaerobes. It was necessary to replace the current culture system with improved system, such as BACTEC 9050 system. The molecular biology technique would make up for the default of ELISA.


Subject(s)
Humans , Bacteremia , Epidemiology , Blood Specimen Collection , Fetal Blood , Microbiology , Virology , Fungemia , Epidemiology , Polymerase Chain Reaction , Probability , Viremia , Epidemiology
17.
Chinese Journal of Pathophysiology ; (12)1986.
Article in Chinese | WPRIM | ID: wpr-674067

ABSTRACT

AIM: To purify human yolk sac mesenchymal ste m cells (hYS-MSC) and investigate its osteogenic and neurogenic differentiation potentials. METHODS: hYS-MSC were separated from yolk sac and purified via p assage culture. The karyotype of hYS-MSCs was analyzed via G-banded characterist ics. Flow cytometric analysis was used to determine the cell cycle and phenotype of hYS-MSC. The AKP expression of hYS-MSC was also tested. Osteogenic different iation of hYS-MSCs was induced by 10 -8mol/L dexamethasone, 10 mmol/L ?-gl ycerophosphate and 50 mg/L vitamin C. Alizarin red S stain was used for identifi cation of mineralization. ?-mecaptoethanol or salviae miltiorrhizae were used t o induce neurogenic differentiation of hYS-MSCs. The expressions of NSE, NF and GFAP were identified by immunohistochemical method. RESULTS: hYS-MSCs could be purified at passages 2 or 3. The cell cycle analysis suggested that hYS-MSCs showed strong proliferational potentials by which the cells kept normal diploid karyotype during the in vitro cultur e. Flow cytometry showed the phenotype of purified hYS-MSCs was uniformly positi ve for CD29, CD44, CD105, and CD166, and negative for reactivity to antigens CD3 4, CD45, or CD86. hYS-MSCs were weakly but clearly positive in AKP. Osteogenic d ifferentiation was appeared after induction of osteogenic differentiation. hYS-M SCs, which were of spindle shape, uniform in size, were induced to pleomorphism osteoblast-like cells which expressed high level of AKP. Aggregates or nodules w ere formed at day 7 and calcium accumulation was detected by alizarin red S stai ning on day 10 or day 14. Neurogenic differentiation of hYS-MSCs was induced by ?-mecaptoethanol or salviae miltiorrhizae. NSE, NF or GFAP positive cells were detected by immunohistochemical staining. CONCLUSIONS: hYS-MSCs have strong proliferation potential and th e normal diploid karyotype is kept during the in vitro culture. The phenoty pe of hYS-MSCs is coincident with adult hMSCs. hYS-MSCs could be induced to dif ferent iate into osteogenic or neurogenic cells.

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