[Objective]This study was designed to investigate capability of the conditioned media that originated from Renca cells to convert CIM~+ CD25~- T cells into CD4~+ CD25~+ T cells,which can exert immunosuppressive effect on effector T cells in vitro and in vivo.[Methods]The common media were mixed with the conditioned media at different ratios,and fresh enriched CD4~+ CD25~- T cells with MACS were cultured in mixed media for 7 days.At end-point of culture,the cells were collected and detected phenotypes in flow cytometer.Moreover,we detected immunosuppressive effect of converted CD4~+ CD25~+ T cells on effeetor T cells proliferation in one-way mixed lymphocytes reaction by using CCK-8,and we observed survival time and histology of grafts.The delayed type hypersensitivity was determined 14 days after transplantation.[Results]The mixed media could increase ratio of CD4~+ CD25~+ Foxp3~+ T cells in conditioned media ratio-dependent(P＜0.05),compared with control groups,when the mixed media contained no mote than 75% of conditioned media.The converted CD4~+ CD25~+ T cells significantly suppress proliferation of effector T cells in vitro,and prolong survival time of grafts,which were(29.6±1.4)d in converted CD4~+ CD25~+ T cells treated groups(P＜0.05),compared with that in untreated groups(9.8±0.6 d)or PBS treated groups(10.9±0.6 d).Moreover,delayed-type hypersensitivity reaction were conducted at day 14 after transplantation in the recipients,and the results showed that less pad swelling in the group treated with converted CD4~+ CD25~+ T cells than other control groups was found,according to measurement of pad swelling.In addition,progressed to complete necrosis of grafts were exhibited in the mice treated with PBS and untreated mice,whereas better healing of grafts and less lymphocytes infiltration were displayed in the mice treated with converted CD4~+ CD25~+ T cells,which were similar to the mice treated with natural regulatory T cells.[Conclusion]The converted CD4~+ CD25~+ T cells with Renca conditioned media play suppressive role in vitro and in vivo.

Objective To study influences of intervention in pregnant women with positive thyroid autoantibodies on the thyroid function of babies. Methods A total of 55 pregnant women were enrolled with positive thyroid peroxidase antibody (TPOAb) and/or thyroglobulin antibody (TgAb) during prenatal checkup. They were randomly divided into two groups: intervening group( n= 36, newborn group A) was treated with levothyroxine ( L-T4 ), and non-intervening group ( n= 19, newborn group B) was not treated. 30 cases of pregnant women with negative thyroid autoantibodies served as a normal population control group (newborn group N). Serum TSH, TPOAb, TgAb, TT3, TT4, FT3 and FT4 were measured by high-sensitive immunochemiluminescent assay ,and urinary iodine was also examined in the pregnant women. Fetal plasma TSH, TT3, TT4, FT3, and FT4 levels were measured after cutting the umbilical cord from placenta, and repeated measurements were made by 3-4 weeks and 8-10 weeks postpartum. Results At baseline, serum TSH levels of the pregnant women in intervening and nonintervening groups were significantly higher than that in control group ( P＜0.05 ). Non-intervening group had higher TSH and lower TT3, TT4, FT4 compared with the other two groups (P＜0. 05 or P＜0.01 ). The cord blood TSH levels of the neonates in both group B [(7.06 ± 1.31 ) mIU/L] and group A [(6.23 ± 1.26 ) mIU/L] were significantly higher than that of group N [(5.48±1. 17) mIU/L, P＜0.01 and 0. 05]. By 3-4 weeks postpartum,the serum TSH level [(3.21±0.70)mIU/L] in group B was significantly higher than those in group N [(2.72±0.51)mIU/L] and group A [(2.78±0.42) mIU/L, all P＜0.05]. The serum TSH level in group B [(2.99±0.57) mIU/L] was still higher than those in group N [(2.48±0.68) mIU/L] by 8 to 10 weeks postpartum (P＜0.05 ). Multiple stepwise regression analysis revealed that TSH, TPOAb, and urine iodine levels of mothers were independently related to TSH of their infants. Conclusion When differences in thyroid function exist in pregnant women, these differences also reside in their offspring. The thyroid function in neonates correlates with both the thyroid autoantibodies and thyroid function of their mothers.

In women with preeclampsia (PE), endothelial cell (EC) dysfunction can lead to altered secretion of paracrine factors that induce peripheral vasoconstriction and proteinuria. This study examined the hypothesis that PE sera may directly or indirectly, through human umbilical vein ECs (HUVECs), stimulate phospholipase C-gamma1-1,4,5-trisphosphate (PLC-gamma1-IP3) signaling, thereby increasing protein kinase C-alpha (PKC-alpha) activity, collagen I expression and intracellular Ca2+ concentrations ([Ca2+]i) in human umbilical artery smooth muscle cells (HUASMCs). HUASMCs and HUVECs were cocultured with normal or PE sera before PLC-gamma1 silencing. Increased PLC-gamma1 and IP3 receptor (IP3R) phosphorylation was observed in cocultured HUASMCs stimulated with PE sera (P<0.05). In addition, PE serum significantly increased HUASMC viability and reduced their apoptosis (P<0.05); these effects were abrogated with PLC-gamma1 silencing. Compared with normal sera, PE sera increased [Ca2+]i in cocultured HUASMCs (P<0.05), which was inhibited by PLC-gamma1 and IP3R silencing. Finally, PE sera-induced PKC-alpha activity and collagen I expression was inhibited by PLC-gamma1 small interfering RNA (siRNA) (P<0.05). These results suggest that vasoactive substances in the PE serum may induce deposition in the extracellular matrix through the activation of PLC-gamma1, which may in turn result in thickening and hardening of the placental vascular wall, placental blood supply shortage, fetal hypoxia-ischemia and intrauterine growth retardation or intrauterine fetal death. PE sera increased [Ca2+]i and induced PKC-alpha activation and collagen I expression in cocultured HUASMCs via the PLC-gamma1 pathway.
Adult ; Apoptosis ; Calcium/*metabolism ; Cell Line ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Collagen Type I/analysis/*metabolism ; Female ; Human Umbilical Vein Endothelial Cells ; Humans ; Muscle, Smooth, Vascular/*cytology/metabolism ; Phospholipase C gamma/genetics/*metabolism ; Pre-Eclampsia/*blood/*metabolism/pathology ; Pregnancy ; Protein Kinase C-alpha/metabolism ; RNA Interference ; *Signal Transduction ; Young Adult

Background::The potential impact of β cell function and insulin sensitivity on adverse pregnancy outcomes in women with gestational diabetes mellitus (GDM) remains uncertain. We aimed to investigate the association between β cell dysfunction, insulin resistance, and the composite adverse pregnancy outcomes.Methods::This observational study included 482 women diagnosed with GDM during pregnancy. Quantitative metrics on β cell function and insulin sensitivity during pregnancy were calculated using traditional equations. The association of β cell dysfunction and insulin resistance with the risk of the composite adverse pregnancy outcomes was investigated using multivariable-adjusted logistic regression models.Results::Multivariable-adjusted odds ratios (ORs) of adverse pregnancy outcomes across quartiles of homeostatic model assessment for insulin resistance (HOMA-IR) were 1.00, 0.95, 1.34, and 2.25, respectively ( P for trend = 0.011). When HOMA-IR was considered as a continuous variable, the multivariable-adjusted OR of adverse pregnancy outcomes was 1.34 (95% confidence interval 1.16-1.56) for each 1-unit increase in HOMA-IR. Multivariable-adjusted ORs of adverse pregnancy outcomes across quartiles of homeostatic model assessment for β cell function (HOMA-β) were 1.00, 0.51, 0.60, and 0.53, respectively ( P for trend = 0.068). When HOMA-β was considered as a continuous variable, the multivariable-adjusted OR of adverse pregnancy outcomes was 0.57 (95% CI 0.24-0.90) for each 1-unit increase in HOMA-β. However, other quantitative metrics were not associated with the composite adverse pregnancy outcomes. Conclusions::We demonstrated a significant association of β cell function and insulin sensitivity with the risk of adverse pregnancy outcomes. We have provided additional evidence on the early identification of adverse pregnancy outcomes besides the glycemic values.