Arterial PO2, PCO2, and pH versus transcutaneous PO2 and PCO2 and tissue pH in the fetal dog

In the fetal dog, simultaneous recording by transcutaneous PO2 and PCO2 and tissue pH electrodes were compared to corresponding arterial values during hypoxic episodes produced by occlusion of the maternal abdominal aorta. Before occlusion, the differences between the paired values were minimal. Under anoxic conditions, expected changes in the peripheral circulation were observed. However, the transcutaneous PO2 was lower, the transcutaneous PCO2 much higher, and the tissue pH much lower than in blood. Continuous electrodes demonstrate changes resulting from gas and hydrogen ion coming from cells more readily than blood because they are closer to the former. Additionally, carbon dioxide and hydrogen ion are buffered to a greater degree in blood than in cells. Consequently, under conditions of stress and active metabolism, PCO2 is higher while PO2 and pH are lower in cells than in blood. When compared with monitoring of gases and acid-base balance through blood sampling, continuous transcutaneous and intracutaneous monitoring would seem to be more representative of the environment at the cellular level.     

Continuous intrapartum pH, pO2, pCO2, and SpO2 monitoring

The goal of intrapartum surveillance and its further development is better patient care for both the fetus and the gravida. A normal FHR pattern is usually associated with the delivery of a normal well-oxygenated infant; however, a nonreassuring FHR is not always associated with the delivery of a compromised infant. This situation has led to an increase in unnecessary obstetric interventions in the form of a rising cesarean section rate. Fetal scalp sampling was developed in an attempt to improve the predictive value of electronic FHR monitoring, but because this technique is not widely used, management decisions are frequently made using FHR patterns alone. Much research has been performed in the search for a continuous biochemical measurement of fetal status, including continuous pH, pO2, or pCO2 and various combinations of these methodologies. None of these measurements are used in current clinical practice, mainly owing to technical problems and difficulties associated with the continuous direct measurement of these parameters in fetal blood throughout labor. The promising new field of fetal pulse oximetry has the potential to provide reliable, meaningful, and reproducible data as shown in early cross-sectional studies and more recent longitudinal studies. By identifying developing hypoxia, this technology may reduce the uncertainty associated with electronic FHR monitoring. Fetal pulse oximetry may also provide critical information relating to the detection and management of the hypoxic fetus. Any new method of intrapartum fetal monitoring requires careful evaluation to assess its potential value before its introduction into clinical practice. The use of fetal SpO2 monitoring in the presence of a nonreassuring FHR pattern is being examined in a multicenter randomized controlled trial. This study will address the question of whether supplementary monitoring of fetal SpO2 levels can lead to a reduction in the cesarean section rate for fetal distress. The available data on fetal noninvasive pulse oximetry have been obtained from a combination of well-designed cohort studies (level II-2 evidence) or from earlier multiple time series (level II-3 evidence). The results from the US Multicenter Trial (level I evidence) should provide a significant addition to current evidence. A continuous fetal noninvasive monitor measuring fetal oxygenation directly could lead to an improvement in the sensitivity and specificity of fetal surveillance. This improvement could ultimately result in a reduction in unnecessary interventions by differentiating hypoxic fetuses from nonhypoxic fetuses and, more importantly, may lead to earlier intervention for fetuses in danger of serious compromise.     

LMTK2 and PARP-2 gene polymorphism and azoospermia secondary to meiotic arrest

Purpose

To investigate whether the human LMTK2 and PARP-2 gene defects are associated with azoospermia by meiotic arrest, mutational analysis was performed on Japanese men with azoospermia.

Methods

Via direct sequencing, mutational screening was carried out on the exon region of the genes, using genomic DNAs from 18 Japanese men. Statistical analysis was done on the detected single nucleotide polymorphisms (SNPs) in the patients and normal controls.

Results

Nine SNPs were detected in LMTK2 and five SNPs were detected in PARP-2. There were no significant differences in the genotype distribution and allele frequencies between the two groups in LMTK2. However, the genotype frequency of heterozygotes in SNP1 of PARP-2 was higher in the patient group. The haplotype analysis revealed that SNP1-SNP4 (T-A) of PARP-2 was significantly more frequent in the patient group.

Conclusion

The PARP-2 gene might be associated with azoospermia by meiotic arrest in humans.     

Macrophage-activating lipopeptide-2 induces cyclooxygenase-2 and prostaglandin E(2) via toll-like receptor 2 in human placental trophoblast cells

We have examined whether toll-like receptor (TLR)2-mediated stimulation by macrophage-activating lipopeptide-2 (MALP-2), originally purified from Mycoplasma fermentans, induces cyclooxygenase (COX)-2 and prostaglandin (PG)E(2) in human placental trophoblast cells. The signaling mechanism by which MALP-2 exerts its effect has also been examined. Human placental trophoblast cells isolated from term placenta were used. TLR expression in trophoblast cells was confirmed by multiplex PCR and immunocytochemistry, and examined whether MALP-2 induces COX-2 and PGE(2) by Northern blotting, RT-PCR, Western blotting and ELISA, respectively. The activation of NF-kappaB and MAP kinases (ERK1/2 and p38) was examined by Western blotting. The effects of inhibitors of NF-kappaB, MEK1/2 and p38 on MALP-2-induced PGE(2) production were also evaluated. TLR2, TLR6 and TLR4 were expressed in human placental trophoblast cells. MALP-2 significantly induced COX-2 expression and enhanced PGE(2) production in a dose-dependent manner. MALP-2 induced the activation of NF-kappaB, ERK1/2 and p38 MAPK. Inhibitors of NF-kappaB, MEK1/2 and p38 blocked MALP-2-inducible PGE(2) production. TLR2-mediated stimulation by MALP-2 induces COX-2 and PGE(2) in human placental trophoblast cells via NF-kappaB and MAP kinases pathways.     

Combined intracervical PGE2 and intra-amniotic PGF2 alpha for induction of 2nd trimester abortion

Fourteen consecutive patients (mean gestational age 18.1 weeks, range 15-23 weeks) referred for therapeutic termination of pregnancy were induced into abortion by intra-amniotic PGF2 alpha 40 mg followed by oxytocin stimulation. 14 other patients (mean gestational age 17.9 weeks, range 15-23 weeks) were pretreated with intracervical PGE2 1.0 mg in gel for 4 h prior to induction of abortion with intra-amniotic PGF2 alpha 40 mg without further stimulation. The induction-abortion interval for patients treated with intra-amniotic PGF2 alpha and oxytocin, was 19.1 +/- 2.94 h (+/- SE, n = 14) with a success rate of 80% after 24 h. After pretreatment with intracervical PGE2 1.0 mg in viscous gel, intra-amniotic PGF2 alpha 40 mg induced abortion after 11.2 +/- 1.12 h (+/- SE, n = 14) with a 100% success rate after 24 h. No systemic side effects of the PGE2 pretreatment were noted. No cervical laceration was observed. The results need further confirmation, but still suggest cervical priming with intracervical PGE2 1.0 mg in gel and subsequent induction of abortion by intra-amniotic PGF2 alpha 40 mg as an attractive principle for 2nd trimester abortion.     

Expression of gelatinase (MMP-2 and MMP-9) and cyclooxygenase-2 (COX-2) in endometrial carcinoma

OBJECTIVE: Matrix metalloproteinases (MMPs) are key players in the degradation of extracellular matrix and basement membranes, and are thus important in tumor invasion. Gelatinases (MMP-2 and MMP-9) in particular are prognostic factors in many solid tumors. In this study the immunohistochemical expression of both COX-2 and matrix metalloproteinases has been shown for the first time in endometrium carcinoma. METHODS: Forty-two endometrial carcinoma tissues were immunostained for MMP2 antibody (1:100, Rabbit polyclonal), MMP9 antibody (1:100, Rabbit polyclonal) and CoX2 antibody (1:100, Epitope specific rabbit antibody). RESULTS: 90.5% of the cases were positive for MMP-2 and MMP-9, and 83.3% of the cases were positive for COX-2. A statistically significant association was found between COX-2 overexpression and FIGO stage (p = 0.001). A positive correlation was also found with histological grade (p = 0.006), myometrial invasion (p = 0.033), vascular invasion (p = 0.017), and lymphatic invasion (p = 0.007). A positive correlation was found between MMP-2 overexpression and vascular and lymphatic invasion (p = 0.030 and p = 0.003, respectively). MMP-9 overexpression was also found to be correlated with vascular and lymphatic invasion (p = 0.001 and p = 0.012, respectively). Furthermore, there was a statistically significant correlation between MMP-2 and MMP-9 overexpression (p = 0.0001). CONCLUSION: The results showed that COX-2, MMP-2 and MMP-9 were expressed in a high percentage of primary endometrial carcinomas and their expressions may be associated closely with parameters of tumor aggressiveness.     

环氧合酶-2、bcl-2蛋白在宫颈鳞癌组织中的表达及意义

目的:探讨宫颈鳞癌组织中环氧合酶(COX-2)、bcl-2蛋白的表达及其与宫颈癌临床分期、病理分级、淋巴转移的关系。方法:采用免疫组化法对40例宫颈癌组织中COX-2和bcl-2蛋白的表达进行检测分析,以10例正常宫颈组织作为对照。结果:COX-2和bcl-2蛋白在正常宫颈组织中阳性表达率均为0(0/10)。COX-2在宫颈原位癌、Ⅰ~Ⅱ期及Ⅲ期的阳性表达率分别为50.0%、54.5%和66.7%,宫颈鳞癌及正常宫颈组织组之间比较差异有显著性(P<0.05),宫颈原位癌、宫颈鳞癌Ⅰ、Ⅱ、Ⅲ期间差异无显著性(P>0.05)。COX-2在Ⅰ~Ⅱ期宫颈鳞癌表达与淋巴结转移有关(P<0.05)。bcl-2在宫颈原位癌、宫颈鳞癌Ⅰ~Ⅱ期、Ⅲ期癌细胞胞浆中未着色,但在癌间质有不同程度染色。bcl-2在宫颈癌及正常宫颈组织组之间差异有显著性(P<0.05)。宫颈原位癌、宫颈癌Ⅰ、Ⅱ、Ⅲ期间差异无显著性(P>0.05)。bcl-2与癌细胞分化程度及淋巴结转移无关(P>0.05)。COX-2与bcl-2在宫颈鳞癌组织中表达无相关性(P>0.05)。结论:COX-2和bcl-2蛋白在正常宫颈组织中均不表达。COX-2在宫颈鳞癌中的表达与临床分期和病理分级无关,与淋巴结转移有关。二者在宫颈癌中的表达无相关性。检测宫颈鳞癌组织中COX-2的表达有利于早期诊断及估计宫颈癌患者的预后。     

Analysis of GM2 and anti-GM2 antibody in normal pregnancy

Oncofetal antigen immunogenic (OFA-I), an antigen common to the fetal brain and some malignant tumors (derived from ectoderm), is immunogenic in man, and anti-OFA-I antibody appears in the sera of not only malignant patients but also pregnant women. Biochemical analysis revealed that OFA-I is ganglioside GM2 and GD2. In the present study, GM2 and anti-GM2 antibody were investigated as to the kinetics in pregnant sera and the tissue localization in placenta and decidua. The results obtained were as follows: 1. The concentration of GM2 was 76.7 pmol/ml (mean) in pregnant sera and 10.8 pmol/ml (mean) in cord sera. GM2 was not measurable in the sera of non-pregnant women. 2. GM2 was detected by thin layer chromatogram-immunostaining methods in the gangliosides extracted from decidua and term placenta. GM2 was, however, undetected from villi at an early stage. 3. In immunohistological analysis for GM2, positive findings were observed in decidua and in villous stroma of term placenta. In contrast, no distinct positive findings were observed in trophoblast at any stage. 4. Anti-GM2 antibody appeared in pregnant sera at an early stage and was detectable up to the 5th day postpartum.     

Prostaglandin F2 alpha and E2 release by peritoneum with and without endometriosis

Peritoneal endometriotic implants and adjacent normal peritoneum from five patients were analyzed for prostaglandin (PG) release. Each tissue biopsy was incubated using medium 199 in triplicate at 37 degrees C for six hours, and PGF2 alpha and PGE2 concentrations were measured in the incubation medium every two hours. This study demonstrates that peritoneum involved with endometriosis releases significantly more PGF2 alpha and PGE2 (P less than .05) than adjacent normal peritoneum, and suggests that peritoneal endometriotic implants may be a source of the elevated peritoneal fluid PG levels previously reported in patients with endometriosis.     

Human placental oxytocin and its increase by prostaglandin E2 and F2 alpha

To investigate the physiological roles of human placental oxytocin (OT)-like substance, partial purification of human placental extract and placental tissue culture were carried out. 1. Both oxytocic immunoreaction in our own specific radioimmunoassay and bioaction on rat uterine contraction were observed in the larger (MW greater than or equal to 5,000) and the smaller molecular part (MW not equal to 1,000) in partially purified placental extracts by gel-filtration using Sephadex G-25. 2. Prostaglandin E2 and F2 alpha enhanced the immunoreactive OT in the cultured human term placental tissue at doses of 1-100 micrograms/ml. These findings are considered to be significant in basic studies of human placental OT-like substance.     

Cyclooxygenase-2 derived PGE2 and PGI2 play an important role via EP2 and PPARdelta receptors in early steps of oil induced decidualization in mice

Differentiation of endometrial stromal cells into decidual cells (decidualization) is prerequisite for blastocyst implantation. Different prostanoids are shown to be involved in the cascade of events found in implantation and decidualization. Previous reports described that cyclooxygenase-2 (COX2) derived prostacyclin (PGI2) plays an important role via peroxisome proliferator activated receptor (PPARdelta) nuclear receptor in implantation and decidualization. Herein, we investigated the role of COX2 derived PGE2 and PGI2 and examined the protein expression and regulation of COX1, COX2, membrane-bound prostaglandin E synthase (mPGES-1), prostaglandin I synthase (PGIS), PGE2 receptor (EP2) and PPARdelta in hormone primed oil infused uterine horn as well as in non-infused uterine horn (control horn). Our results show that selective COX2 inhibitor (Nimesulide) inhibits decidualization while COX1 inhibitor (SC560) does not affect decidualization. COX2, mPGES-1, PGIS, EP2 and PPARdelta immunostaining are strongly observed at 24 h and 48 h in oil-induced horn and than significantly reduced at 72 h and 120 h and absent in non-infused horn. However COX1 immunostaining is observed in infused as well as in non-infused horn. Our immunohistochemical studies corroborated well with follow up western blotting of the same proteins. PGE2 and PGI2 products were also elevated at 24h and 48 h after oil induction in infused horn in comparison to control horn. Our data suggest that COX2 derived both PGE2 and PGI2 mediate its function via EP2 and PPARdelta receptors in early steps of decidualization in mice.     

PGE2与胎儿胎盘循环

目的　探讨前列腺素Ｅ２（ＰＧＥ２ ）对正常妊娠及妊高征孕妇胎儿胎盘循环的调节作用。　方法　在５０例足月正常孕妇（对照组）及３７例妊高征合并ＩＵＧＲ孕妇（妊高征组）中,利用共焦点激光扫描显微镜测定了ＰＧＥ２对人脐静脉内皮细胞（ＨＵＶＥＣ）内游离钙离子（Ｃａ２＋ ）ｉ动态变化的影响,同时用放免法测定了其脐静脉血中ＰＧＥ２浓度。　结果　ＰＧＥ２ 使两组ＨＵＶＥＣ中（Ｃａ２＋ ）ｉ上升, 脐血中ＰＧＥ２ 值在妊高征组为５．４±０．６ ｐｇ／ｍ ｌ与对照组的１５．３±１．２ ｐｇ／ｍ ｌ相比,显示有意义的低值（Ｐ＜０．０５）。　结论　ＰＧＥ２ 可能为胎儿胎盘循环的血管扩张因子;妊高征组脐血中ＰＧＥ２ 浓度降低使胎儿胎盘血流阻力增加可能影响胎儿生长发育     

NS-398与前列腺素E_2对子宫内膜异位症子宫内膜细胞环氧合酶-2mRNA表达与凋亡的影响

目的 :探讨选择性环氧合酶 2 (COX- 2 )抑制剂NS -398与前列腺素E(PGE2 )对子宫内膜异位症患者子宫内膜细胞COX -2mRNA表达与细胞凋亡的影响。方法 :以体外培养的子宫内膜细胞为研究对象 ,分别用NS- 398与PGE2 处理。采用RT- PCR法、MTT法、酶联免疫吸附试验 (ELISA)和流式细胞术 ,检测刺激前后COX -2mRNA表达量、细胞增殖、凋亡与细胞周期分布情况以及上清液中凋亡抑制蛋白Bcl 2与PGE2 的释放量。结果 :NS- 398以剂量与时间依赖方式抑制COX -2mRNA的表达以及PGE2 与Bcl -2的分泌 ,抑制子宫内膜细胞增殖 ,诱导细胞凋亡 ,改变细胞周期分布 ,增加G0 /G1期细胞的比例。PGE2以时间与剂量依赖方式刺激子宫内膜细胞的COX- 2mRNA的表达 ,使Bcl- 2释放增加。同时 ,PGE2 可以逆转NS- 398对子宫内膜细胞的抑制作用 ,细胞增殖重新活跃 ,改变细胞周期分布 ,减少G0 /G1期细胞的比例 ,抑制细胞凋亡。结论 :COX- 2选择性抑制剂NS- 398,促进细胞凋亡 ,抑制细胞增殖 ,其机制可能与抑制COX -2的表达 ,降低PGE2 以及Bcl- 2释放 ,和改变细胞周期有关。PGE2 在体外能够刺激子宫内膜细胞COX- 2的表达升高 ,促进细胞增殖 ,抑制细胞凋亡。