To evaluate the role of nitric oxide in local function of endometrium, the pattern of expression of endothelial and inducible nitric oxide synthases in the human endometrium at proliferative and secretory phases and decidua was studied by using immunocytochemistry. Results showed: (1) At the proliferative phase, the eNOS immunostaining was confined to vascular endothelial, whereas the iNOS was not detected in any composition of endometrium. (2) At the secretory phase, surface epithelium and grandular epithelium showed positive staining for both eNOS and iNOS. The stroma remained uniformly negative throughout the menstrual cycle. (3) In the decidua, the expression of both isoforms was increased, while moderate iNOS immunoreactivity was observed in decidualized stromal cells. It was demonstrated that the expression of eNOS and iNOS was elevated at the secretory phase and in decidua, indicating the increased production of NO at these phases. The increasing of NO might take part in implantation through dilating the vessels and relaxing the smooth muscles and in menstration through promoting apoptosis.

To evaluate the role of nitric oxide in the embryo implantation in mice, three experiments were carried out using the mice implantation model. In the experimentⅠand experimentⅡ, NG-nitro-L-arginine methyl ester (L-NAME), the non-specific NOS inhibitor, was administered intraperitoneally at doses of 0.5 mg-5 mg on the day 3 of pregnancy and at dose of 3 mg on different days of pregnancy (day 1-6) . In the experiment Ⅲ, L-aragnine, a donor of NO, was co-administered with L-NAME to evaluate the effect of L-aragnine on L-NAME, and the embryo development was assessed. In all these three experiments, the endometrium was histologically examined. Results showed compared with the control groups intraperitoneal administration of a dose of L-NAME between 1 and 5 mg on the day 3 of pregnancy led to the decrease in the number of implantation sites (P<0.05), and 4 to 5 mg of L-NAME caused inhibition of implantation completely. L-NAME resulted in failure of pregnancy when administrated at 3 mg between day 3 and 5 of pregnancy. The characteristic vascular permeability changes and decidualization in the endometrium were significantly attenuated and embryo growth was retarded. The L-NAME-mediated effects were significantly reversed when L-aragnine was co-administered with L-NAME. This study demonstrated that NO promoted the implantation in mice through regulating the permeability and decidulization of endometrium and the development of embryo.

In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kunming mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.
Blastocyst/cytology ; Embryo Implantation ; Endometrium/*metabolism ; Gene Expression ; Gene Expression Regulation, Developmental ; Leukemia Inhibitory Factor/*biosynthesis ; Leukemia Inhibitory Factor/*genetics ; Medicine, Chinese Traditional ; Models, Biological ; Plant Extracts/pharmacology ; RNA, Messenger/metabolism ; Time Factors

The debate exists whether or not gonadotropin-releasing hormone (GnRH) analogs used in controlled ovarian hyperstimulation (COH) impair endometrial receptivity. Homeobox A11 (Hoxa11), Meis homeobox 1 (Meis1), cadherin 1 (Cdh1), and catenin beta 1 (Ctnnb1) are well known to be involved in successful implantation. In this study, the endometrial expression of Hoxa11, Meis1, Cdh1, and Ctnnb1 during the peri-implantation period was investigated in an in vitro fertilization (IVF) mouse model by real-time RT-PCR and Western blot to evaluate the relationship between Hoxa11, Meis1, Cdh1, and Ctnnb1 expression and the impact of the COH on endometrial receptivity. The mimic COH protocols included GnRH agonist plus human menopausal gonadotropin (HMG) (GnRH agonist group), GnRH antagonist plus HMG (GnRH antagonist group), and HMG alone (HMG group). The expression levels of Hoxa11, Meis1, Cdh1, and Ctnnb1 mRNA and protein were decreased in all of the COH groups. The expression levels of Hoxa11 and Ctnnb1 were the lowest in the GnRH agonist group, and those of Meis1 and Cdh1 were lower in the GnRH analog groups than the HMG group. There were positive correlations between the expression of Hoxa11 and Ctnnb1, as well as the expression of Meis1 and Cdh1 among all the groups. In conclusion, the COH protocols, particularly with GnRH analogs, suppressed Hoxa11, Meis1, Ctnnb1 and Cdh1 expression, in mouse endometrium during the peri-implantation period. Our data reveal a novel molecular mechanism by which the COH protocols might impair endometrial receptivity.

Objective To investigate the effect of sepsis on vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.Methods Thirty-six adult male SPF SpragueDawley rats,aged 2-3 months,weighing 200-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:control group (group C),sham operation group (group S) and sepsis group (group Sep).Sepsis was induced by cecum ligation and puncture (CLP) in rats anesthetized with intraperitoneal chloral hydrate 350 mg/kg.At 12 h after CLP,the sciatic nerve-pretibial muscle was prepared.Vecuronium was added to the culture medium with the final concentration of 0.08 μg/ml,and the sciatic nerve-pretibial muscle was incubated for 15 min.Before and after administration,evoked endplate potentials (EPPs) and miniature endplate potentiais (MEPPs) were recorded by using intracellular microelectrode.EPP/MEPP ratio was calculated.Results Compared to C and S groups,EPPs,MEPPs and EPP/MEPP ratio were significantly increased before and after administration in group Sep.EPPs,MEPPs and EPP/MEPP ratio were significantly lower after administration than before administration in the three groups.Conclusion Sepsis can promote acetylcholine release in neuromuscular junction,thus weakening vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.

Objective To evaluate the effects of unilastatin on brain injury in children undergoing aortic arch surgery under cardiopulmonary bypass (CPB).Methods Twenty ASA physical status Ⅲ or Ⅳ children of both sexes,aged 1-24 months,weighing 3-12 kg,undergoing repair of coarctation of aorta or interrupted aortic arch complicated with intracardiac malformations under CPB,were randomly divided into 2 groups (n =10 each):control group (group C) and ulinastatin group (group U).Ulinastatin 20 000 U/kg was diluted into 10 000 U/ml with normal saline and it was then injected intravenously in 3 parts (1/3 was injected via the internal jugular vein after induction of anesthesia; 1/3 at the beginning of CPB and 1/3 at 5 min before aortic unclamping).In group C the equal volume of normal saline was given instead of ulinastatin.Blood samples were taken from the radial artery after induction of anesthesia (T1),at 10 min after aortic clamping (T2),at 10 min after aortic unclamping (T3),at the end of CPB (T4),and at 6 and 24 h after termination of CPB (T5,T6) for determination of plasma S100B protein and neuron-specific enolase (NSE) concentrations.Results There was no significant difference in plasma levels of S100B protein and NSE at T1 between the two groups (P ＞ 0.05).Plasma S100B protein and NSE levels were significantly increased at T2-5 as compared to the baseline values at T1 in both groups (P ＜ 0.05).Plasma S100B protein and NSE levels were significantly lower at T2-5 in group U than in group C (P ＜ 0.05).Conclusion Ulinastatin can attenuate brain injury in children undergoing aortic arch surgery under CPB.

In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kun- ruing mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of L1F mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve em- bryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.

The debate exists whether or not gonadotropin-releasing hormone (GnRH) analogs used in controlled ovarian hyperstimulation (COH) impair endometrial receptivity.Homeobox A11 (Hoxall),Meis homeobox 1 (Meisl),cadherin 1 (Cdhl),and catenin beta 1 (Ctnnbl) are well known to be involved in successful implantation.In this study,the endometrial expression of Hoxall,Meisl,Cdhl,and Ctnnbl during the peri-implantation period was investigated in an in vitro fertilization (IVF) mouse model by real-time RT-PCR and Western blot to evaluate the relationship between Hoxall,Meisl,Cdhl,and Ctnnbl expression and the impact of the COH on endometrial receptivity.The mimic COH protocols included GnRH agonist plus human menopausal gonadotropin (HMG) (GnRH agonist group),GnRH antagonist plus HMG (GnRH antagonist group),and HMG alone (HMG group).The expression levels of Hoxall,Meisl,Cdhl,and Ctnnbl mRNA and protein were decreased in all of the COH groups.The expression levels of Hoxall and Ctnnbl were the lowest in the GnRH agonist group,and those of Meisl and Cdbl were lower in the GnRH analog groups than the HMG group.There were positive correlations between the expression of Hoxall and Ctnnbl,as well as the expression of Meisl and Cdhl among all the groups.In conclusion,the COH protocols,particularly with GnRH analogs,suppressed Hoxall,Meisl,Ctnnbl and Cdhl expression,in mouse endometrium during the peri-implantation period.Our data reveal a novel molecular mechanism by which the COH protocols might impair endometrial receptivity.