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Objective@#To investigate the proportion and role of CD45+ erythroid progenitor cells (EPC) in patients with tongue cancer metastasis.@*Methods@#The initial treatment of tongue cancer patients (n=40) from January 2017 to June 2018 in He'nan Provincial People′s Hospital was included in this study. According to the presence or absence of lymph node metastasis, they were divided into tumor group (no lymph node metastasis was found in imaging and pathology) and metastasis group (both imaging and pathology confirmed lymph node metastasis). The expression of Ki-67 was detected by immunohistochemistry and the proportion of CD45+CD71+TER119+EPC was detected by flow cytometry. EPC was sorted by flow cytometry, interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) were detected by enzyme-linked immunosorbent assay (ELISA), and reactive oxygen species (ROS) was detected by flow cytometry. Transwell was used for tumor invasion test; methyl thiazolyltetrazolium (MTT) assay was used to detect proliferation level.@*Results@#There were 20 cases in the tumor group and metastasis group. There was no significant difference between the two groups in terms of age, sex, time of onset and size of tumors. Flow cytometry showed that the ratio of CD45+EPC in peripheral blood of tumor group and metastasis group was (1.2±0.2)% and (3.1±0.2)% (t=7.823, P<0.001). Correlation analysis showed that the ratio of CD45+EPC was positively correlated with the proliferation index of Ki-67 cells (r=0.592, P=0.006). The results of flow cytometry showed that the mean fluorescence intensity (MFI) of ROS in EPC was 102.1±22.9 in tumor group and 530.0±67.2 in metastasis group (t=6.025,P<0.001). The results of ELISA showed that the mass concentrations of IL-10 and TGF-β in EPC supernatant of tumor group were (10.8±1.6) and (3.2±0.8) μg/L, respectively. The mass concentrations of IL-10 and TGF-beta in EPC supernatant of metastasis group were (26.9±3.7) and (6.4±0.9) μg/L, respectively (t=3.956, P=0.003; t=2.595, P=0.027). Transwell results showed that the proportion of invasive cells in the CD45+EPC group [(40.3±4.4)%] was higher than that in the control group [(17.5±2.2)%] (t=4.607, P=0.001). MTT proliferation experiment showed that the proliferation rate of the CD45+EPC group [(52.0±3.3)%] was higher than that of the control group [(30.5±1.9)%] (t=5.656, P<0.001).@*Conclusions@#The proportion of CD45+EPC in patients with tongue cancer metastasis is significantly increased. CD45+EPC can promote the proliferation and metastasis of tongue cancer by secreting immunosuppressive molecules and ROS.
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Objective:To study the effects of hyaluronic acid(HA) and TGF-β1 on the growth of mandibular condylar cartilage and the hyperthophic differentiation of the condylar chondrocyts.Methods:60 condyle samples from newborn mice were in vitro cultured and treated with HA(0.5 mg/ml),TGF-beta 1 (5 ng/ml) and without additional agent(the control) respectively.The Morphological observation,Alizarin Red Staining,Alkaline phosphatase staining and condylar cartilage surface area measurement were conducted after 1,2,4,6 and 8 weeks of culture respectively.Results:High-density photoresist area was observed in the condylar cartilage of the control group after 4 weeks of culture.Alizarin Red Staining and Alkaline phosphatase staining showed condylar cartilage matrix production and calcification.The HA group showed no high-density photoresist area at all time points,however,the cartilage area was significantly increased (P ＜ 0.05);the TGF-beta 1 group showed high-density photoresist area after 2 weeks of culture.but the cartilage area were not significantly changed(P ＞ 0.05).Conclusion:HA can promote the growth of condylar cartilage in vitro,but have an inhibitory effect on chondrocyte differentiation.TGF-β1 plays a role in mandibular condylar chondrocyte hypertrophic differentiation in the early days of in vitro culture.
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Objective:To study the difference of the development between mandibular condylar cartilage and femoral head cartilage in vitro.Methods:Mandibular condyles and femoral heads were sampled from 1 2 neonatal mice and cultured in vitro.The samples before culture and after 6-week culture were examined by gross observation,HE staining,Alizarin Red staining and PCNA immunohistochem-istry respectively.Results:After in vitro culture,abnormal changes were observed in condyle cartilage,but the surface area of condyle cartilage was not changed(P >0.05).HE staining showed partial cartilage layer structure disappearance and the Alizarin Red staining confirmed calcification in the cartilage matrix.However,calcification was not found in femoral head cartilage,and the surface area of femoral head cartilage increased(P <0.05).HE staining showed the hypertrophied layer was thicker after culture than before and the Alizarin Red staining showed there was no calcification in the femoral cartilage matrix.The immunohistochemistry displayed PCNA posi-tive expression in both cartilage after culture.Conclusion:In vitro,the mandibular condylar cartilage matrix can be spontaneously cal-cificated.