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OBJECTIVES@#To explore the molecular characteristics of Staphylococcus aureus (S. aureus) in children, and to compare the molecular characteristics of different types of strains (infection and colonization strains) so as to reveal pathogenic molecular markers of S. aureus.@*METHODS@#A cross-sectional study design was used to conduct nasopharyngeal swab sampling from healthy children in the community and clinical samples from infected children in the hospital. Whole genome sequencing was used to detect antibiotic resistance genes and virulence genes. A random forest method to used to screen pathogenic markers.@*RESULTS@#A total of 512 S. aureus strains were detected, including 272 infection strains and 240 colonization strains. For virulence genes, the carrying rates of enterotoxin genes (seb and sep), extracellular enzyme coding genes (splA, splB, splE and edinC), leukocytotoxin genes (lukD, lukE, lukF-PV and lukS-PV) and epidermal exfoliating genes (eta and etb) in infection strains were higher than those in colonization strains. But the carrying rates of enterotoxin genes (sec, sec3, seg, seh, sei, sel, sem, sen, seo and seu) were lower in infection strains than in colonization strains (P<0.05). For antibiotic resistance genes, the carrying rates of lnuA, lnuG, aadD, tetK and dfrG were significantly higher in infection strains than in colonization strains (P<0.05). The accuracy of cross-validation of the random forest model for screening pathogenic markers of S. aureus before and after screening was 69% and 68%, respectively, and the area under the curve was 0.75 and 0.70, respectively. The random forest model finally screened out 16 pathogenic markers (sem, etb, splE, sep, ser, mecA, lnuA, sea, blaZ, cat(pC233), blaTEm-1A, aph(3')-III, ermB, ermA, ant(9)-Ia and ant(6)-Ia). The top five variables in the variable importance ranking were sem (OR=0.40), etb (OR=3.95), splE (OR=1.68), sep (OR=3.97), and ser (OR=1.68).@*CONCLUSIONS@#The random forest model can screen out pathogenic markers of S. aureus and exhibits a superior predictive performance, providing genetic evidence for tracing highly pathogenic S. aureus and conducting precise targeted interventions.
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Enfant , Humains , Staphylococcus aureus/génétique , Études transversales , Entérotoxines/génétique , Infections à staphylocoques , Séquençage du génome entierRésumé
An 80-year-old male patient was hospitalized with tremor.Through the detailed collection of medical history and perfect examination,it was determined that the disease was caused by lasidipine nimodipine,tremor disappeared after withdrawal,and no recurrence after 1 year follow-up.Although tremor does not affect life,it directly affects the quality of life of patients,and the awareness and attention of this adverse reaction should be improved clinically.
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OBJECTIVE@#To investigate the clinical efficacy of high dose methotrexate (HD-MTX), temozolomide (TMZ), and rituximab (R) in the treatment of patients with primary central nervous system lymphoma (PCNSL).@*METHODS@#Clinical data of patients with PCNSL diagnosed and treated in Guangdong Provincial People's Hospital from February 2010 to May 2017 were collected. First, patients were given 6-8 cycles of MTX (3.5 g/m@*RESULTS@#There were 42 patients enrolled in the study, 17 cases in HD-MTX+TMZ group and 25 cases in HD-MTX+TMZ+R group. The median PFS and OS times in HD-MTX+TMZ+R group were 56.7 months and N/A, respectively, while, 7.3 months and 34.7 months in HD-MTX+TMZ group, respectively. In addition, there was no significant difference in median survival between patients who received TMZ maintenance therapy and those who were only actively monitored. During the induction period, all the patients had grade 1-2 nausea and vomiting, while in the consolidation treatment period, no grade 3/4 toxicity was observed.@*CONCLUSION@#The combination of HD-MTX+TMZ+R in the treatment of PCNSL patients shows a definite short-term effect, which can increase the survival rate of the patients. The side effects are mild, and the patients can generally tolerate.
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Humains , Protocoles de polychimiothérapie antinéoplasique , Système nerveux central , Tumeurs du système nerveux central/traitement médicamenteux , Lymphome malin non hodgkinien/traitement médicamenteux , Méthotrexate/usage thérapeutique , Études rétrospectives , Rituximab/usage thérapeutique , Témozolomide/usage thérapeutique , Résultat thérapeutiqueRésumé
<p><b>BACKGROUND</b>The prognostic values of interim and post-therapy fluorine-18-fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) and PET/computed tomography (CT) scanning have been confirmed in several subtypes of lymphoma. However, its prognostic value in Burkitt's lymphoma has not been clearly defined. The aim of the present study was to assess the prognostic value of PET/CT scanning during different treatment processes of Burkitt's lymphoma.</p><p><b>METHODS</b>A total of 29 adult patients with newly diagnosed Burkitt's lymphoma were retrospectively involved in this study; of them, 23 patients underwent baseline PET/CT, 15 patients underwent mid-therapy PET/CT after 1-4 cycles of chemotherapy, and 17 patients underwent post-therapy PET/CT after all planned first-line chemotherapy cycles. Mid-therapy and post-therapy PET/CT results (positive vs. negative) were visually interpreted according to the criteria of the International Harmonization Project. The reduction in the maximum standardizes uptake values (∆SUVmax) of 25%, 50%, and 75% were regarded as cutoff points. Overall survival (OS) and progression-free survival (PFS) were regarded as the major endpoints.</p><p><b>RESULTS</b>The median OS and PFS were 27.6 months (range 6.5-78.3 months) and 27.2 months (range 3.0-78.3 months), respectively. The median SUVmax of the baseline PET/CT was 18.3 (range 1.6-35.9), whereas the median SUVmax of the mid-therapy and post-therapy PET/CT decreased to 4.0 (range 0-17.6) and 3.0 (range 0-14.5), respectively. The patients' Eastern Cooperative Oncology Group (ECOG) scores (<2 vs. ≥2) were significantly associated with the baseline PET/CT SUVmax. The mid-therapy and post-therapy PET/CT results (positive vs. negative) showed no significant association with OS or PFS. The optimal cutoff ∆SUVmax from the baseline to the post-therapy PET/CT that could predict a change in OS in patients with Burkitt's lymphoma was 50% (P = 0.019).</p><p><b>CONCLUSIONS</b>(18)F-FDG uptake was intense in Burkitt's lymphoma, and there was a significant reduction in SUVmax during the interim and post-therapy PET/CT procedures. A ∆SUVmax of greater than 50% was a favorable cutoff point to predict the OS of Burkitt's lymphoma patients.</p>
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Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Protocoles de polychimiothérapie antinéoplasique , Utilisations thérapeutiques , Lymphome de Burkitt , Imagerie diagnostique , Traitement médicamenteux , Fluorodésoxyglucose F18 , Estimation de Kaplan-Meier , Tomographie par émission de positons couplée à la tomodensitométrie , Méthodes , Pronostic , Radiopharmaceutiques , Études rétrospectives , Résultat thérapeutiqueRésumé
<p><b>OBJECTIVE</b>To observe the changes of the mechanical pain threshold in the rat model of autoimmune prostatitis, explore the mechanism of autoimmune prostatitis pain and offer some animal experimental evidence for the drug therapy of the condition.</p><p><b>METHODS</b>Twenty male Wistar rats weighing 180 - 220 g were divided into a model and a control group. The autoimmune prostatitis model was established by subcutaneous injection of an extract of male rat prostate glands (RPG) at 60 mg/ml in Freund's complete adjuvant (FCA) and pertussis-diphtheria-tetanus vaccine at 0 and 30 days, respectively. Mechanical tactile hyperalgesia was measured once a week using Von Frey Filaments from the beginning of the study. At 8 weeks after modeling, the rats were sacrificed and the prostate tissues harvested for observation of histomorphological changes by HE staining.</p><p><b>RESULTS</b>HE staining revealed different degrees of benign prostatitis in the model rats. Compared with the controls, the mechanical pain threshold in the model rats was significantly decreased with the increased time of modeling, from (65.52 +/- 6.27) g at 0 week to (23.67 +/- 4.09) g at 8 weeks (P < 0.01). Statistically significant differences were found in the variation trend at different time points between the two groups (P < 0.01).</p><p><b>CONCLUSION</b>Autoimmune prostatitis models were successfully established in rats and hyperalgesia was induced after modeling.</p>
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Animaux , Mâle , Rats , Maladies auto-immunes , Modèles animaux de maladie humaine , Seuil nociceptif , Physiologie , Prostatite , Allergie et immunologie , Rat WistarRésumé
Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) are the major cause of in-stent restenosis (ISR). Intervention proliferation and migration of VSMCs is an important strategy for antirestenotic therapy. Roscovitine, a second-generation cyclin-dependent kinase inhibitor, can inhibit cell cycle of multiple cell types. We studied the effects of roscovitine on cell cycle distribution, proliferation and migration of VSMCs in vitro by flow cytometry, BrdU incorporation and wound healing assay, respectively. Our results showed that roscovitine increased the proportion of G0/G1 phase cells after 12 h (69.57±3.65 vs. 92.50±1.68, P=0.000), 24 h (80.87±2.24 vs. 90.25±0.79, P=0.000) and 48 h (88.08±3.86 vs. 88.87±2.43, P=0.427) as compared with control group. Roscovitine inhibited proliferation and migration of VSMCs in a concentration-dependent way. With the increase of concentration, roscovitine showed increased capacity for growth and migration inhibition. Roscovitine (30 μmol/L) led to an almost complete VSMCs growth and migration arrest. Combined with its low toxicity and selective inhibition to ISR-VSMCs, roscovitine may be a potential drug in the treatment of vascular stenosis diseases and particularly useful in the prevention and treatment of ISR.
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Animaux , Rats , Cycle cellulaire , Lignée cellulaire , Mouvement cellulaire , Occlusion du greffon vasculaire , Traitement médicamenteux , Métabolisme , Anatomopathologie , Muscles lisses vasculaires , Métabolisme , Anatomopathologie , Myocytes du muscle lisse , Métabolisme , Anatomopathologie , Inhibiteurs de protéines kinases , Pharmacologie , Purines , PharmacologieRésumé
Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) are the major cause of in-stent restenosis (ISR). Intervention proliferation and migration of VSMCs is an important strategy for antirestenotic therapy. Roscovitine, a second-generation cyclin-dependent kinase inhibitor, can inhibit cell cycle of multiple cell types. We studied the effects of roscovitine on cell cycle distribution, proliferation and migration of VSMCs in vitro by flow cytometry, BrdU incorporation and wound healing assay, respectively. Our results showed that roscovitine increased the proportion of G0/G1 phase cells after 12 h (69.57±3.65 vs. 92.50±1.68, P=0.000), 24 h (80.87±2.24 vs. 90.25±0.79, P=0.000) and 48 h (88.08±3.86 vs. 88.87±2.43, P=0.427) as compared with control group. Roscovitine inhibited proliferation and migration of VSMCs in a concentration-dependent way. With the increase of concentration, roscovitine showed increased capacity for growth and migration inhibition. Roscovitine (30 μmol/L) led to an almost complete VSMCs growth and migration arrest. Combined with its low toxicity and selective inhibition to ISR-VSMCs, roscovitine may be a potential drug in the treatment of vascular stenosis diseases and particularly useful in the prevention and treatment of ISR.
Résumé
<p><b>OBJECTIVE</b>To explore the relationship between aging and erectile function changes in rats in order to establish a rat model of aging-related erectile dysfunction (ED).</p><p><b>METHODS</b>Eighty male Wistar rats were equally divided into four age groups (3-, 6-, 12- and 18-month) and treated with intragastric administration of sildenafil citrate (Sn) for penile erection tests. Twenty 3-month-old female Wistar rats were randomized to four groups as oestrous rat models. We recorded the rate and frequency of penile erections of the male rats in different age groups.</p><p><b>RESULTS</b>The rates of penile erection were 85%, 75%, 40% and 30% and erectile frequencies were 2.27 +/- 0.80, 2.00 +/- 0.61, 1.40 +/- 0.51 and 1.29 +/- 0.49 in the 3-, 6-, 12- and 18-month rats, respectively, with statistically significant differences among different age groups (P < 0.01). And their erectile function exhibited a tendency to decrease with the increase of age. Besides, comparison of the 3-month with the 6-, 12- and 18-month groups showed significantly reduced erectile function in the 18-month rats (P < 0.05) but no remarkable difference between the 3-month and the 6- and 12-month groups (P > 0.05).</p><p><b>CONCLUSION</b>Aging is one of the main risk factors of rat erectile dysfunction, and 18-month-old male rats are qualified for the establishment of the rat model of aging-related erectile dysfunction.</p>
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Animaux , Mâle , Rats , Vieillissement , Physiologie , Dysfonctionnement érectile , Modèles animaux , Érection du pénis , Physiologie , Rat WistarRésumé
<p><b>OBJECTIVE</b>To investigate the effects of enriched environment and impoverished environment on the learning and memory ability of manganese-exposed mice and the mechanism.</p><p><b>METHODS</b>Forty female Kunming mice were randomly and equally divided into 4 group: control group (CG), standard environment and manganese exposure group (SEG), enriched environment and manganese exposure group (EEG), and impoverished environment and manganese exposure group (IEG). The mouse model of manganese poisoning was established by intraperitoneal injection of manganese chloride. The learning and memory ability was tested by Morris water maze. The expression of cAMP response element-binding protein (CREB) in area CA1 of the hippocampus was measured by immunohistochemistry.</p><p><b>RESULTS</b>In place navigation test, the SEG had a significantly longer escape latency than the CG (P < 0.05), and the EEG had a significantly shorter escape latency than the SEG (P < 0.05); there was no significant difference in escape latency between IEG and SEG (P > 0.05). In spatial probe test, the EEG had a significantly greater number of platform crossings than the SEG (P < 0.05), and the IEG had a significantly smaller number of platform crossings than the SEG (P < 0.05). The expression of CREB in area CA1 of the hippocampus was significantly lower in IEG and SEG than in CG (P < 0.05), and it was significantly higher in EEG than in SEG (P < 0.05).</p><p><b>CONCLUSION</b>In the enriched environment, the learning and memory ability of manganese-exposed mice can be improved, which may be due to the increased expression of CREB in the hippocampus.</p>
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Animaux , Femelle , Souris , Protéine de liaison à l'élément de réponse à l'AMP cyclique , Métabolisme , Modèles animaux de maladie humaine , Environnement , Hippocampe , Métabolisme , Apprentissage , Intoxication au manganèse , Métabolisme , MémoireRésumé
<p><b>OBJECTIVE</b>To investigate the migration of fluorescent dye PKH26-labeled BM-MSC in the Alzheimer's model rats.</p><p><b>METHODS</b>Normal human bone marrow extracted for isolation of BM-MSC was cultured in vitro. The 5th passaged BM-MSC was labeled with PKH26, and observed under a fluorescence microscope for PKH26 labeling efficiency, and using flow cytometry BM-MSC surface markers was checked. The PKH26 labeled BM-MSC injected into the tail vein of the normal control group and AD animal model group, 14 days after finding the PKH26-labeled BM-MSC cells in the rat hippocampus using fluorescence microscopy. Using the Morris water maze experiment comparison of AD model and BM-MSC transplantation group of spatial learning and memory ability.</p><p><b>RESULTS</b>TFlow cytometry showed BM-MSC surface markers CD73 and CD105 were positive. In vitro, PKH26-labeled rate of BM-MSC was 100 %. The Morris water maze experiment comparison of BM-MSC transplantation group and AD group of animals, BM-MSC transplantation group at 13, 14 days of spatial learning and memory ability than AD animal group had significantly improved. 14 days after BM-MSCs in rat hippocampus could be found which were PKH26-positive, consistent with DAPI staining. PKH26-positive cells in animal models of AD were significantly more than those in the normal control group.</p><p><b>CONCLUSION</b>BM-MSC in AD rats not only migrates through the blood-brain barrier, but also mainly survives in the hippocampus of AD rats, and it can improve AD rat model of learning disabilities.</p>
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Animaux , Humains , Mâle , Rats , Maladie d'Alzheimer , Anatomopathologie , Cellules de la moelle osseuse , Biologie cellulaire , Mouvement cellulaire , Cellules cultivées , Modèles animaux de maladie humaine , Injections veineuses , Cellules souches mésenchymateuses , Biologie cellulaire , Composés chimiques organiques , Rat Sprague-DawleyRésumé
<p><b>OBJECTIVE</b>To establish an animal model of chronic nonbacterial prostatitis (CNP) using different doses of purified prostate protein with Freund's complete adjuvant (FCA), and to investigate the relationship of the doses with the success of the model construction.</p><p><b>METHODS</b>Thirty male Wistar rats were divided into a control (A) and 4 experimental groups (B, C, D and E) of equal number. The latter 4 groups were given multi-loci intracutaneous injection of 1.0 ml of a 1:1 mixture of purified prostate protein at 20, 40, 60 and 80 mg/ml with Freund's complete adjuvant (FCA), and meanwhile intraperitoneally injected with 0.5 ml of pertussis-diphtheria-tetanus vaccine at 0 and 30 days. On the 45th day, the rats were sacrificed for observation of the pathomorphological changes in the prostate glands with the naked eyes and microscope.</p><p><b>RESULTS</b>Different degrees of chronic inflammation were observed with different degrees of lymphocyte infiltration and interstitial hyperplasia in the experimental rats. More obvious changes were found in Groups C and D than in A, and even more significant in Group E (P < 0.05).</p><p><b>CONCLUSION</b>The rat model of CNP can be successfully established by multi-loci intracutaneous injection of 1.0 ml of a 1: 1 mixture of purified prostate protein at 40 - 60 mg/ml with FCA, and simultaneously intraperitoneal injection of 0.5 ml of pertussis-diphtheria-tetanus vaccine twice within 30 days.</p>
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Animaux , Mâle , Rats , Auto-immunité , Modèles animaux de maladie humaine , Relation dose-réponse (immunologie) , Adjuvant Freund , Pharmacologie , Prostatite , Rat WistarRésumé
<p><b>OBJECTIVE</b>To investigate the clinicopathological features of EB virus positive diffuse large B-cell lymphomas (EBV + DLBCL) of the elderly.</p><p><b>METHODS</b>Four hundred and ninety-six cases of DLBCLs were retrospectively studied by in situ hybridization (ISH) to detect the EBV in tumor cells, and by immunohistochemistry to evaluate the expression of CD10, CD20, CD30, CD79a, bcl-6, bcl-2, MUM-1, CD5, CD3, TIA-1 and Ki-67 protein. Their clinicopathological correlations were analyzed.</p><p><b>RESULTS</b>Of the 59 cases of EBV + DLBCL, 48 cases were EBV positive. The median age of these EBV + DLBCLs was 73 years with male predominance (1.4:1). There were 11 cases with nodal presentation only, 18 cases with extra-nodal presentation and 19 cases with both lymph nodal and extra-nodal involvements, whereas about one third cases with more than one extra-nodal involvement. Thirty-five patients presented with advanced disease (Ann Arbor stage III/IV). A performance status was available in 36 cases and 5 cases had performance status of more than 1. Seven of 30 patients were found with high lactate dehydrogenase value (more than twice of the normal). An IPI-score was calculated in 30 cases and 18 cases had an intermediate/high IPI-score (3-5). The median survival for these patients was 35 months. Morphologically, EBV + DLBCLs of the elderly generally showed a diffuse and polymorphic proliferation of large lymphoid cells with varying degrees of reactive components including small lymphocytes, plasma cells, histiocytes, and epithelioid cells. These tumor cells were frequently characterized by a broad range of B-cell maturation, containing centroblasts, immunoblasts, and Hodgkin- and Reed-Sternberg (HRS)-like giant cells. The study cohort was further morphologically divided into large cell lymphoma subtypes (n = 33) and polymorphic lymphoma subtypes (n = 14) and one case with mixed subtype. Immunohistochemical studies showed that tumor cells were positive for CD20 (47/48) and/or CD79a (45/45) in almost cases. Tumor cells were MUM-1-positive in the majority of the cases (44/47) and were stained for CD10 or bcl-6 in a few cases. Expression of bcl-2 and CD30 was observed in 80.0% (28/35) and 28.9% (11/38) cases, respectively, and most of the cases (33/39) had a high proliferative index (by Ki-67 with a 50% cut-off point). Compared with other EBV + DLBCLs, except the older age and low frequency of bcl-6 staining, no other significant differences were observed in EBV + DLBCLs of the elderly.</p><p><b>CONCLUSIONS</b>EBV + DLBCLs of the elderly constitute a distinct clinicopathologic subtype of DLBCL, although many clinical and histological features with EBV + lymphomas are similar with that of younger ages. Differential diagnosis from other types of lymphomas should also be considered.</p>
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Sujet âgé , Sujet âgé de 80 ans ou plus , Femelle , Humains , Mâle , Adulte d'âge moyen , Antigènes CD20 , Métabolisme , Protocoles de polychimiothérapie antinéoplasique , Utilisations thérapeutiques , Antigènes CD79 , Métabolisme , Cyclophosphamide , Utilisations thérapeutiques , Doxorubicine , Utilisations thérapeutiques , Infections à virus Epstein-Barr , Études de suivi , Herpèsvirus humain de type 4 , Facteurs de régulation d'interféron , Métabolisme , Antigènes CD30 , Métabolisme , L-Lactate dehydrogenase , Sang , Lymphome B diffus à grandes cellules , Traitement médicamenteux , Métabolisme , Anatomopathologie , Virologie , Stadification tumorale , Prednisone , Utilisations thérapeutiques , Études rétrospectives , Taux de survie , Vincristine , Utilisations thérapeutiquesRésumé
<p><b>OBJECTIVE</b>To investigate the role of bcl-6 gene rearrangement and bcl-6 expression in three molecular subgroups of diffuse large B-cell lymphoma (DLBCL) and its clinicopathological significance.</p><p><b>METHODS</b>Tissue microarray including 163 newly diagnosed DLBCL was constructed. Fluorescence in situ hybridization (FISH) was performed to detect the bcl-6 gene rearrangement and immunohistochemistry (EnVision method) was used to evaluate the expression of bcl-6, Ki-67, cyclin D3, Geminin and P27(Kip1) proteins in DLBCL. The association with clinicopathological features was analyzed.</p><p><b>RESULTS</b>One hundred and forty nine of 163 cases were further classified into three molecular subgroups: 40 cases of germinal center B-cell-like (GCB) type, 75 cases of activated non-germinal center B-cell-like (ABC) type, 34 cases of Type 3. Of these 149 cases, FISH for bcl-6 gene rearrangement was successful in 118 cases. bcl-6 gene rearrangement was observed in 33 of 118 (28.0%) cases. The bcl-6 gene rearrangement was more frequently seen in the ABC subgroup (22/62, 35.5%) than in GCB (6/31, 19.4%) and Type 3 subgroups (5/25, 20.0%, P=0.16). The correlation of bcl-6 gene rearrangement and expression of its encoded protein was further analyzed. Most of DLBCL (26/33, 78.8%) with bcl-6 gene rearrangement presented with overexpression of its encoded protein, which was higher than those without bcl-6 gene rearrangement (53/84, 62.4%, P=0.088). DLBCL with bcl-6 gene rearrangement (24/33, 72.7%) more frequently expressed cyclin D3, and had a higher proliferative activity than those without bcl-6 gene rearrangement (37/81, 45.7% , P=0.009). Twenty-nine of 33 (87.9%) cases of DLBCL with bcl-6 gene rearrangement presented with advanced stage (Ann Arbor stage III/IV), which was higher than those without bcl-6 gene rearrangement (65/85, 76.5% , P=0.167). Univariate Cox proportional hazards regression analysis showed that bcl-6 gene rearrangement was associated with an increased relative risk (at 1.842) of death in DLBCL cases compared with those without bcl-6 gene rearrangement.</p><p><b>CONCLUSION</b>Overexpression of bcl-6 protein caused by bcl-6 gene rearrangement may play some important roles in the development and/or progression of a subset of DLBCL.</p>
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Humains , Lymphocytes B , Anatomopathologie , Chromosomes humains de la paire 14 , Cycline D3 , Génétique , Réarrangement des gènes , Immunohistochimie , Hybridation fluorescente in situ , Lymphome B , Diagnostic , Génétique , Lymphome B diffus à grandes cellules , Diagnostic , Génétique , Métabolisme , Anatomopathologie , Stadification tumorale , Pronostic , Protéines proto-oncogènes c-bcl-6 , Génétique , Translocation génétiqueRésumé
<p><b>OBJECTIVE</b>To investigate the differentiative capability of adult human bone marrow mesenchymal stem cells (BMSCs) into Schwann-like cells.</p><p><b>METHODS</b>Bone marrows were aspirated from healthy donors and mononuclear cells were separated by Percoll lymphocytes separation liquid (1.073 g/ml) with centrifugation, cells were cultured in DMEM/F12 (1:1) medium containing 10% fetal bovine serum (FBS), 20 ng/ml epidermal growth factor (EGF) and 20 ng/ml basic fibroblast growth factor (bFGF). Cells of passage 1 were identified with immunocytochemistry.</p><p><b>RESULTS</b>Mononuclear cells separated by Percoll's were passaged 10 times by trypsin/ethylenediaminetetraacetic acid (EDTA) digestion in 40 days, and BMSCs increased about 6x10(7) times in this short period. Immunohistochemistry identified that BMSCs were CD34- and CD31-, but they expressed neuron specific enolase; 0.01%-0.02% of total cells expressed nestin, the marker for neural progenitor cells; 40%-50% cells stained heavily by neurofilament 200; and no glial fibrillary acidic protein (GFAP) positive cells were identified; S100 expression was detected among 0.1%-0.2% cells.</p><p><b>CONCLUSIONS</b>Bone marrow contains the stem cells with the ability of differentiating into Schwann-like cells, which may represent an alternative stem cell sources for neural transplantation.</p>
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Adulte , Humains , Cellules de la moelle osseuse , Biologie cellulaire , Métabolisme , Différenciation cellulaire , Physiologie , Prolifération cellulaire , Immunohistochimie , Protéines de filaments intermédiaires , Métabolisme , Cellules souches mésenchymateuses , Biologie cellulaire , Métabolisme , Protéines de tissu nerveux , Métabolisme , Nestine , Protéines neurofilamenteuses , Métabolisme , Enolase , Métabolisme , Protéines S100 , Métabolisme , Cellules de Schwann , Biologie cellulaireRésumé
To explore the effect of antisense phosphorothioate oligodeoxynucleotide (ASODN) of human telomerase reverse transcriptase (hTERT) gene on telomerase activity in CEM cells, PCR enzyme-linked immunoassay was used to determine telomerase activity. The expression levels of hTERT mRNA and protein were assayed by RT-PCR and immunofluorescence assay using fluoresce isothiocyanate label respectively. The results showed that the expression levels of hTERT mRNA and protein in CEM cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT mRNA and protein levels between control and sense oligodeoxynucleotide-treated cells. Telomerase activity decreased when CEM cells were treated with ASODN for 48 hours. Telomerase activity of CEM cells was significantly inhibited when treated with ASODN for 72 hours. There was no difference in telomerase activity levels between control and hTERT sense oligodeoxynucleotide-treated cells. These results suggested that hTERT ASODN inhibited telomerase activity of CEM cells.
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Humains , Division cellulaire , Lignée cellulaire , Protéines de liaison à l'ADN , Cytométrie en flux , Oligodésoxyribonucléotides antisens , Pharmacologie , ARN messager , Telomerase , Génétique , MétabolismeRésumé
An embryo of 4.3 mm is described. It comes to the laboratory in 10 per cent for-malin as an intact chorion measuring 22 by 16 mm. The embryo is in the shape of theletter C, and its tail is curved. The limb buds are round in shape. There are 4pharyngeal pouches, the 4th being very small. Between the bases of the 1st and 2ndbranchial arches, in the floor of the pharynx, the tuberculum impar has developed. Theotic vesicles are slightly elongated sacs and no longer connected with the covering ecto-derm. The eyes are represented by the optic vesicles and the thickened ectodermal an-lage of the lens. Trachea is still largely a groove in the ventral wall of the pharynx andesophagus. The heart is still in the so-called S shaped stage; the atrium lies on the leftside of the bulb, and has not yet doubled; the sinus venosus is not completely out ofthe septum transversum; in the atrium, near the sinus venosus, the endothelial tube isclosely attached to the wall. There are 4 pairs of aortic arches: the lst pair breaks intocapillaries imbedded in the mandible. The other three join the dorsal aortae which runcaudal and finally unite to form one median dorsal aorta. The anterior cardinal, pos-terior cardinal, common cardinal and vitelline veins, etc. are all paired and symetricallyarranged. The embryo herein described falls in group Ⅷ of Streeter.