| PART ICLINICAL OBSERVATION OF DIFFERENT HAARTREGIMENS ON HIV-INFECTED PREGNANTWOMEN AND HIV-UNINFECTED NEWBORNSObjective: To explore the effectiveness and safety of pregnant women andnewborns exposed to highly active antiretroviral therapy (HAART) duringpregnancy; to observe the pregnant results and changes of maternal or fetalhematopoiesis and immune function in two different regimen of HAART.Methods: According to the different HAART regimens,71HIV-infectedpregnant women with single birth were divided into2groups,40ofcontaining zidovudine (AZT) group（AZT group）,31of AZT-free group,40healthy pregnant women in the control group. The antiretroviral therapy wasbegun in or after the14th pregnant week. The HAART regimens of the AZTgroup were AZT+3TC+LPV/r or+EFV or+NVP, the regimens of theAZT-free group were TDF+3TC+LPV/r or+EFV or+NVP. The samples of venous blood of the maternal and umbilical cord blood were collected indifferent points of time respectively, before antiretroviral treatment, beforedelivery and after delivery. The clinical research information as fellows:①the basic clinical information of the pregnant women and newborns;②hematologic parameters;③CD4+, CD8+T lymphocytes in peripheral blood;④the index of HIV nucleic acid was detected in4to6weeks and4monthsafter birth. Results:(1) Among two groups of the HAART, there were nogestational hypertension and gestational diabetes, and there were no birthdefects, stillbirth and MTCT in newborns.(2) No severe neonatal asphyxiaoccurred, and there were not statistically significant difference in mildasphyxia among the three groups (P>0.05). There were not significantdifferences between two therapy groups in umbilical cord blood levels of Tlymphocytes. The birth weights of newborns exposed to HAART were lowerthan the control group (P <0.05). The low birth weight rate were15.00%,16.13%and5.00%in the three groups, the rates of the two therapy groupswere higher than the control group (P <0.05).(3) The median duration oftreatment in AZT group was20weeks, while in the AZT-free group was21weeks; before the treatment, there were no significant differences between twotreatment groups in CD4+T lymphocyte count and CD4/CD8rate. But beforedelivery, CD4+T lymphocyte count and CD4/CD8in the AZT group weresignificantly higher than the AZT-free group (P <0.05), CD4+was560.32±226.10cells/mm3and CD4/CD8rate was0.63±0.29in AZT grouprespectively. Before delivery, the CD4+T lymphocyte of the AZT group wassignificantly higher than that before therapy, the mean increased and reachedto129.79cells/mm3, the increase was30.15%(P <0.05). While CD8+Tlymphocyte was significant lower than that before therapy， the mean decreased and reached to415.97cells/mm3，the decrease was29.57%(P <0.05). In addition, the rate of CD4/CD8increased significantly (P＜0.05). Inthe AZT-free group, there were no such significant changes of lymphocyteindexes in the two points of time.(4) Before delivery, the hematologicparameters of pregnant women with megaloblastic anemia in the AZT groupcompared with it of the control group, the statistical results as fellows: RBC,Hb, HCT and N decreased (P <0.05), MCV, MCH and RDW increased (P <0.05). Among the three groups, there were no significant differences in WBC,PLT and mild anemia (P>0.05); but in AZT group, the rate of moderateanemia reached to17.50%, it was more significant higher than other groups (P<0.05).(5) Among three groups, there were significant differences in RBC,Hb, HCT, MCV, MCH, RDW, N, L and PLT of umbilical cord blood ofnewborns (P <0.01), but there was no significant difference in WBC. In theAZT group, Megaloblastic anemia of newborns occurred easily; comparedwith the control group, the changes of hematologic parameters: RBC, Hb,HCT and N decreased (P <0.05), but MCV, MCH, RDW and L increased (P<0.05). In the two groups exposed to HAART, the rates of mild anemia were52.50%and29.03%and more significant higher than that of the control group(P <0.05). In the AZT group，the rate of moderate anemia was17.50%, andmore significant higher than that in the AZT-free group (P <0.05).Conclusion:①The MTCT rate of HIV was the lower-level because of a longterm HAART treatment, it show that there are enough effectiveness and safetyin HAART.②In the state of immune reconstruction, the long term AZ-containing HAART regimen is better than AZT-free regimen.③Duringpregnancy, maternal and neonatal megaloblastic anemia occur easily innewborns and pregnancy women with AZT-containing HAART exposure. PART ⅡRESEARCH ON INHIBITIONS OF OFFSPRINGHEMATOPOIETIC STEM/PROGENITOR CELLACTIVITY CAUSED BY ZIDOVUDINE DURINGPREGNANCYObjectives: To observe the suppressions of hematopoietic stem/progenitorcell activity of newborn and newborn mice caused by the exposure ofzidovudine(AZT). Methods: The research of zidovudine intervention onhematopoietic stem/progenitor cell activity:①The clinical research: thesamples of umbilical cord blood of newborns exposed to HAART werecollected；the indexes of hematopoietic stem/progenitor cell activity asfellows: the proportion of CD34+cells detected by flow cytometer (FCM) andthe colony-forming colonies of granulocytes monophyletic progenitor cells(CFU-GM), erythroid progenitor cells (BFU-E) and megakaryocyte progenitorcells (CFU-Meg).②AZT intervention in vitro: umbilical cord bloodhematopoietic stem/progenitor cells and bone marrow cells of newborn micewere isolated and cultured in vitro. The concentrations of AZT were0.1μM,0.5μM,1.0μM,2.0μM and4.0μM respectively. The indexes of hematopoieticstem/progenitor cell activity as fellows: the colony-forming colonies ofCFU-GM, BFU-E, CFU-Meg and apoptosis rates of hematopoietic stem/ progenitor cells detected by FCM.③The pregnancy Balb/c mice wereexposed to AZT with the dose of5.0mg/day and10.0mg/day, exposureperiods, from the10thday of pregnancy to birth. The colony-forming coloniesof CFU-GM, BFU-E, CFU-Meg were measured in monocytes of bone marrowcell of newborn mice. Results: In15cases of cord blood of newborn ofpregnancy women exposed to HAART, the positive rate of CD34+cells was22.6±9.6%and the CFU-GM, BFU-E and CFU-Meg were significantlylower than normal levels (P<0.05). Above the concentrations of0.5μM AZT,the number of colonies in the CFU-GM, BFU-E and CFU-Meg of bonemarrow cells of newborn mice and hematopoietic stem/progenitor cells ofumbilical cord blood were significantly reduced, while the rates of apoptosiswas significantly higher than control in vitro. In the concentration of0.1μMAZT, the rate of apoptosis was not significantly changed, the CFU-GMcolonies were significantly reduced, and the BFU-E and the CFU-Megcolonies decreased, but the differences were not significant. In different dosesof AZT, the CFU-GM, the BFU-E and the CFU-Meg colonies of newbornswere significant less than normal levels in vivo. Conclusions: In thenewborns of pregnancy mice exposed to AZT, the activity and apoptosis ofhematopoietic stem/progenitor cells are inhibited and increased respectively.It suggests that mitochondrial toxicity of hematopoietic stem/progenitor cellscaused by AZT. PART ⅢRESEARCH ON SELF-RENEWAL SIGNALINGPATHWAY OF OFFSPRING HEMATOPOIETIC STEMCELL REGULATED BY ZIDOVUDINEObjectives: To investigate the regulation of self-renewal signaling pathwayPI3K-AKT-mTOR and Bmi-1gen of hematopoietic stem/progenitor cells, innewborns of pregnancy mice exposed to Zidovudine (AZT). Methods: Thepregnancy Balb/c mice were exposed to AZT with the dose of5.0mg/day and10.0mg/day, exposure periods, from the10thday of pregnancy to birth. Theoffspring hematopoietic stem/progenitor cells of bone marrows of wereisolated by Ficoll. The self-renewal signaling pathways PI3K/Akt/mTOR,Bmi-1and PTEN gene expression were measured by qRT-PCR and Westernblot. In addition, the control group was normal newborn mice, adult mice andaged mice. Results: In newborn mice of pregnancy Balb/c mice exposed toAZT, the mTOR expression levels of offspring hematopoietic stem/progenitor cells increased, but had no significant differences with adult mice,and were significantly less than the aged mice; while PTEN gene expressionwere not significant changes; Bmi-1gene expression level were more higherthan the normal newborn mice and adult mice, but were significantly less thanthe aged mice. The changes of protein expressions of P70S6,4EBP1D ofoffspring hematopoietic stem/progenitor cells as follows: newborn mice <newborn mice-AZT≈adult mice <aged mice. Conclusions: In offspringhematopoietic stem/progenitor cells，the limited up-regulation of mTOR ofself-renewal signaling pathway and self-renewal gen Bmi-1are caused by pregnancy mice exposed to AZT, but it can locate in normal range. |
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