Objective To explore the correlation between the amount of residual undifferentiated embryonic stem cells (ESCs) in embryoid bodies and its tumorigenicity.Methods Mouse R1 ESCs were cultured in suspension to form embryoid bodies (EBs).Ten days later,EBs were digested into single cells and then re-plated in standard ESCs culture condition.The residual undifferentiated embryonic stem cells surface maker SSEA-1 was examined by flow cytometry in EBs.The morphology of residual undifferentiated cells in EBs were observed,meanwhile the surface marker SSEA-1 was examined by immunofluorescent staining.EBs were digested into single cells and grouped into 104,105,106,2×106,and then injected into limb muscle of nude mice.The correlation of the amount of cells and its tumorigenicity was observed.Results Residual undifferentiated ESCs were observed after EBs differentiated for 10 days,which displayed clonal morphology and expressed undifferentiated ceil markers of ESCs,such as SSEA-1.The expression rate of undifferentiated cells surface marker SSEA-1 was (13.5±0.75)％ in EBs differentiated for 10 days.Only two millions single cells harvested from EBs were able to form teratoma after being injected into muscle of nude mice for 6 weeks.Mature endoderm,mesoderm and ectoderm tissues could be found in teratoma.No teratoma formed in other groups.Conclusion A certain amount residual undifferentiated ESCs still exist after differentiation of ESCs into EBs.About 2.7× 105 undifferentiated cells are able to form teratoma by iniecting into muscle of nude mice.

Objective To investigate the significance of interleukin-6 IL-6),interleukin-8 (IL-8) and metal matrix protease-9 (MMP-9) in lung injury caused by intrauterine infection in ueonatal rats.Method Eighteen 18-day-old SD pregnant rats were selected and randomly assigned into 0.6,0.7,0.8,0.9,1.0 mg/kg lipopolysaccharide (LPS) groups and control group.Rats of LPS group were injected intraperitoneally with LPS 0.6.0.7,0.8,0.9,1.0 mg/kg,those of control group was injected with equal volume of sterile saline.After 24 hours of injection,one pregnant rat from each group underwent cesarean seetion and placentae were collected for pathological evaluation.The other pregnant rats delivered spontaneously.When the neonatal rats came to 1,3,and 7 days of age,8 rats were randomly selected from each group.The respiratory frequency,body weight,lung weight,and blood samples for detecting inflammatory factors using enzyme-linked immunosorbent assay were measured among 3-day-old neonatal rats.Lung tissue for hematoxylin-eosin staining and lung pathology score were taken in the 1,3 and 7-day-old neonatal rats.Result (1) The mortality rate of neonatal rats increased gradually with the increasing dose of LPS injection.The mortality rate of neonatal rats in the LPS 1.0 mg/kg group was 20％.The placental pathology score and the frequency of 3-day-old neonatal rats in LPS 0.8,0.9,and 1.0 mg/kg groups were higher than control group and other LPS groups (P ＜ 0.05).(2) The histopathological scores of the neonatal rats in the control group and LPS 0.6,0.7,0.8,0.9,1.0 mg/kg groups on the first day after birth were (0.08±0.08),(0.17±0.11),(1.33±0.19),(1.75±0.33),(1.92±0.36),(2.67±0.14),respectively,and that of the third day after birth were (0.00 ± 0.00),(0.50 ± 0.15),(0.67 ±0.19),(1.33 ±0.36),(2.17 ±0.21),(2.92 ±0.08),respectively.LPS 1.0 mg/kg group on the 1st and 3rd day after birth were significantly different from the other groups (P ＜0.05),LPS 0.8 mg/kg group on the 1st and 3rd day after birth were significantly different from the control group and LPS 0.6 mg/kg group (P ＜ 0.05),and there was significant difference among the LPS 0.9 mg/kg group and the other groups on the 3rd day after birth (P ＜ 0.05).When the concentration of LPS was less than 0.8 mg/kg,there was no significant difference in the pathological scores of the lung tissues between each group (P ＞0.05).(3) The concentrations of IL-6,IL-8,and MMP-9 in the neonatal rats of LPS 0.7,0.8,0.9,and 1.0 mg/kg groups were significantly higher than those in the control group at 3 days of age,and the difference among LPS groups were statistically significant (P ＜ 0.05).Conclusion The intraperitoneal injection of LPS in rats 18 days after gestation can lead to lung injury,pulmonary edema,and increasing level of blood IL-6,IL-8,and MMP-9 in the neonatal rats,and this damage lasts until 7 days after birth which suggests the important meaning for lung diseases caused by intrauterine infection.

Objective To investigate the relationship between histological chorioamnionitis (HCA),funisitis (FV) and lung injury in preterm infants.Method Data of preterm infants with gestational age of less than 34 weeks,who were born in our Hospital and admitted to the neonatal ward during the period of October 2015 to August 2016 were retrospectively collected.According to reports of placenta pathologies,participants were assigned into three groups,non-infection group,HCA group and HCA + FV group.Demographic and infectious characteristics,duration of respiratory support,partial pressure of oxygen (PaO2) and pulmonary function testing results at corrected gestational age 40 weeks were compared among groups.Result A total of 151 preterm infants were included,with 33 cases in noninfection group,61 cases in HCA group and 57 cases in HCA + FV group.Non-invasive ventilation duration and number of days in room air in HCA + FV groups were both longer than those in the non-infection group [0 (0,66) h vs.0 (0,0) h,65 (0,176) h vs.0 (0,8) h].The duration of invasive ventilation and total ventilation days in HCA + FV group were longer than those of the other two groups [0 (0,4) h vs.0 (0,0) and 0 (0,0) h,0 (0,99) h vs.0 (0,0) and 0 (0,52) h],the differences were statistically significant (P ＜ 0.05).However,there was no significant difference of PaO2 after 0.5 hour respiratory support among the three groups (P ＞ 0.05).Pulmonary functional parameters,such as respiratory frequency,VPEF/TE,VPEF/VE,VPEF and TPEF,gradually decreased with the degree of inflammatory progress,but there was no statistically significant difference (P ＞ 0.05).Conclusion HCA and FV are highly associated with premature lung injury.The worse the placental inflammation is,the longer respiratory support duration will be needed or required.However,HCA and FV may have no significant effect on lung function.