Initial-onset symptoms of preeclampsia and their relation to pregnancy outcomes.

Preeclampsia (PE) is a serious gestational idiopathic hypertensive disease, threatening both maternal and foetal safety. As a systemic disease, the initial-onset symptoms (IOSs) and clinical manifestations of PE can vary widely from patient to patient. However, a lack of evidence-based data on IOS and their relationship to their corresponding clinical features and pregnancy outcomes persists. We hypothesised that there would be a significant difference between the morbidity time, subsequent organ dysfunction and the status of mother and foetus in PE patients with different IOS. Moreover, early identification of the characteristics of the PE patients with different IOS could improve pregnancy outcomes through individualised prevention or intervention. This study aimed to analyse maternal and foetal condition and pregnancy outcomes of PE patients with different IOS, and to explore the disease progression and characteristics of maternal and foetal outcomes for different IOS, so as to provide the basis for future maternal and foetal monitoring of PE patients.Impact statementWhat is already known on this subject? In 2013, the American College of Obstetricians and Gynecologists revised their definition of PE, sparking a heated debate. Subsequently in 2015, China updated its guidelines to define PE as hypertensive pregnancy accompanied by involvement of any other organ or organ system, to include the heart, lungs, liver and kidneys, among others. However, IOS can be varied in PE, so the maternal management and foetal monitoring should be classified through different IOS. No evidence-based data on IOS in PE patients exist.What the results of this study add? Significant differences in mean morbidity times and mean delivery times were demonstrated among patients with different IOS; medians of the interval from morbidity to delivery were between 4 and 6 weeks. Significant differences in laboratory values were found in patients with different IOS. In patients that did not present with proteinuria as an IOS, 89.1% experienced proteinuria following diagnosis. Patients with the most severe complications presented with hypertension as an IOS. Follow-up visits demonstrated different foetal weight medians.What the implications are of these findings for clinical practice and/or further research? IOS could be an indicator to help evaluate the potential for different maternal and foetal complications and PE outcomes. Moreover, the duration of treatment for PE maybe 4-6 weeks.

Neuromedin B and its receptor influence the activity of myometrial primary cells in vitro through regulation of Il6 expression via the Rela/p65 pathway in mice.

The neuromedin B receptor (Nmbr) is an important physiological regulator of spontaneous activities and stress responses through different cascades as well as its autocrine and paracrine effects. Previous studies have revealed that neuromedin B (Nmb) and its receptor signal via the Rela (also known as p65)/Il6 pathway in a mouse model of pregnancy. This study investigated the mechanism of Nmbr signaling via the Rela/p65-Il6 pathway and regulation of the concentration of intracellular free calcium ([Ca(2+)](i)) during the onset of labor in primary mouse myometrial cell cultures isolated from mice in term labor. Data demonstrated Nmbr agonist-mediated upregulation of the DNA binding activity of Rela/p65, Il6 expression, and [Ca(2+)](i) in a concentration-dependent manner. Furthermore, a significant correlation was observed between DNA binding activity of Rela/p65 and Il6 expression. Moreover, this up-regulation was blocked by Nmbr and Rela/p65 knockdown, achieved by RNA interference (RNAi) technology. No significant differences were identified in the inhibition of Il6 expression as a result of Nmbr or Rela/p65 knockdown. However, significant differences were observed between the [Ca(2+)](i) in Rela/p65-specific group and that in the Nmbr-specific small interfering RNA (siRNA)-treated groups. These data demonstrated that the Nmb/Nmbr interaction in pregnant myometrial primary cells in vitro predominantly influenced uterine activity through regulation of Il6 expression via the Rela/p65 pathway, although the effects of Nmbr on [Ca(2+)](i) involved several pathways that remain to be elucidated.

Neuromedin B and its receptor induce labor onset and are associated with the RELA (NFKB P65)/IL6 pathway in pregnant mice.

Although the neuromedin B receptor (NMBR), a bombesin receptor family member, has been implicated in thermoregulation and in stimulation of both urogenital and gastrointestinal smooth muscle contraction, its underlying role in labor onset and its associated molecular mechanisms remain poorly understood. We examined the relationship between temporal and spatial NMBR expression in the myometrium of pregnant mice and potential mechanistic pathways leading to labor onset. Resultant data indicate that NMBR expression peaked at term and before parturition. Maternal exposure to the NMBR agonist neuromedin B (NMB) shortened the gestational age of pups, an effect that was also observed after oxytocin administration. Both RELA (NFKB P65) DNA-binding activity and interleukin 6 (Il6) mRNA expression were greatest during parturition and after maternal exposure to the highest NMB concentration administered (150 µg/kg). Furthermore, a significant correlation was observed among NMBR mRNA expression, RELA DNA-binding activity, and Il6 mRNA expression. These data demonstrate that NMB and its receptor can induce the onset of labor via a RELA/IL6-mediated pathway.

[The application and significance in prenatal diagnosis using G-banding, fluorescence in situ hybridization and comparative genomic hybridization].

OBJECTIVE: To investigate the procedure and the value of G-banding, fluorescence in sit hybridization (FISH) and comparative genomic hybridization (CGH) techniques in prenatal diagnosis. METHODS: Karyotype analyses with three diagnostic procedures, G-banding, G-banding and FISH, G-banding, FISH and CGH, were performed in the amniotic fluid samples taken from 102 fetuses at gestational ages 16-24 weeks. And the significance was valued in prenatal diagnosis. RESULTS: In the first procedure of karyotype analysis, 98 cases were diagnosed, 2 cases were not conformed while 2 cases were failed in all 102 cases. In the second procedure, 2 cases were determined, 1 case was not conformed and 1 case was still failed. In the third step, 2 cases were diagnosed. The diagnostic rate of the karyotype reached to 100% (102/102 cases) using all the three procedures. In total, seven cases with chromosomal abnormality were diagnosed. Four cases, 1 case and 2 cases were identified in the first step (4/7, 57.1%), the second (1/7, 14.3%) and the third (2/7, 28.5%), respectively. CONCLUSION: It can help improve the diagnostic rate of chromosomal aberrations and standardize diagnostic procedure to perform the three detecting steps in prenatal diagnosis.

[Systematic continuous sequence approach in diagnosing fetal deformity].

OBJECTIVE: To explore the value of ultrasonographic evaluation in fetal deformity in prenatal diagnosis by a systematic continuous sequence approach (SCSA). METHODS: Successive prenatal ultrasonographic evaluation was performed to monitor the whole anatomic structure,form, posture and movement of 16,685 fetuses during gestation aging 14 approximately 40(+3) weeks. RESULTS: Satisfactory ultrasonic images were obtained in 16,627 fetuses using the SCSA (99.65%). Of them, 514 abnormal fetuses were confirmed after subsequent labor or induced labor and 498 abnormal fetuses were correctly diagnosed using SCSA during prenatal stage (96.89%). Whereas 16 fetuses missed recognition (3.11%). Its sensitivity, specificity, positive and negative predictive value of diagnosis on fetal deformity were 96.98%, 99.96%, 98.66%, and 99.90 %, respectively. CONCLUSION: SCSA in prenatal ultrasonographic evaluation of the fetal structure and malformation is reliable and accurate.

Exosomes from human umbilical cord blood accelerate cutaneous wound healing through miR-21-3p-mediated promotion of angiogenesis and fibroblast function.

The application of blood plasma for soft tissue wound healing is receiving much more attention recently. Exosomes are critical paracrine mediators that can be obtained from biological fluids including plasma and be able to induce regenerative effects by transferring bioactive molecules such as microRNAs (miRNAs). This study aimed to investigate the effects of exosomes from human umbilical cord blood plasma (UCB-Exos) on wound healing and to elucidate the underlying mechanism. Methods: UCB-Exos were isolated by ultracentrifugation and subcutaneously injected into full-thickness skin wounds in mice. The efficacy of UCB-Exos on wound healing was evaluated by measuring wound closure rates, histological analysis and immunofluorescence examinations. In vitro, quantitative real-time PCR (qRT-PCR) analysis was performed to detect the expression levels of a class of miRNAs that have positive roles in regulating wound healing. The scratch wound assay, transwell assay and cell counting kit-8 analysis were conducted to assess the effects of UCB-Exos on migration and proliferation of human skin fibroblasts and endothelial cells. Tube formation assay was carried out to test the impact of UCB-Exos on angiogenic tube formation ability of endothelial cells. Meanwhile, by using specific RNA inhibitors or siRNAs, the roles of the candidate miRNA and its target genes in UCB-Exos-induced regulation of function of fibroblasts and endothelial cells were assessed. Results: The local transplantation of UCB-Exos into mouse skin wounds resulted in accelerated re-epithelialization, reduced scar widths, and enhanced angiogenesis. In vitro, UCB-Exos could promote the proliferation and migration of fibroblasts, and enhance the angiogenic activities of endothelial cells. Notably, miR-21-3p was found to be highly enriched in UCB-Exos and served as a critical mediator in UCB-Exos -induced regulatory effects through inhibition of phosphatase and tensin homolog (PTEN) and sprouty homolog 1 (SPRY1). Conclusion: Our results suggest that UCB-Exos are important effectors of plasma activity and can be used as a novel promising strategy for soft tissue wound healing.

[Scanning of drug targets related to uterus contraction from the uterine smooth muscles by cDNA microarray].

OBJECTIVE: To screen the differentially expressed gene profile from the smooth muscles in the fundus uterus at the active stage of labor, and to provide candidate genes for picking out the drug targets related to uterine contraction. METHODS: Differentially expressed genes of uterine smooth muscles in the corpus from pro and post spontaneous parturition and those induced by oxytocin,as well as those from the corpus and the lower portion spontaneous parturition,were scanned respectively by human full-length genetic cDNA microarray with 8064 probe sets. Semi-quantitative RT-PCR was applied to testify the expression of voltage dependent calcium channel-L subtype (CACNA). The differentially expressed genes in the structure and function of the drug targets were picked out by bio-informatics to serve as candidate drug targets related to uterine contraction. RESULTS: The expressions of 29 genes were upregulated in fundus smooth muscles from the pro and post natural parturition, the pro and post inductive parturition of oxytocin, and the natural parturition. The expression of CACNA gene in RT-PCR was in accordance with that in the microarray. Among the 29 genes, neuromedin B receptor (NMBR) gene and neuropeptide Y (NPY) gene were the genes which not only had the targets of uterine contracted medicine, but also could contract the uterine. The differential expression ratios of NMBR in the above 3 types of uterine myometrium were 6.9,11.3, and 9.0, respectively while those of NPY were 6.0,29.8, and 2.9 respectively. CONCLUSION: NMBR, whose expression in the uterine smooth muscles is always up-regulated at different parturition conditions, is likely to be an ideal candidate target of uterotonic drugs.

A new method for selective reduction in TRAP sequence with a contraindication to fetoscopic surgery: A case report.

RATIONALE: The preferred method for multifetal pregnancy reduction (MFPR) is a transabdominal intrathoracic or intracranial injection of potassium chloride (KCl). However, in monochorionic multiple pregnancies (MMPs), especially in monoamnionic multifetal pregnancies, selective feticide by this method is often associated with miscarriage of the remaining fetuses. Selective fetal reduction in MMPs by blood flow ablation using radiofrequency ablation or fetoscopic laser surgery may improve survival of the remaining fetus. Although often successful, MFPR by these methods is contraindicated in cases of twin reversed arterial perfusion (TRAP) sequence in triplet pregnancies complicated by polyhydramnios or anterior placenta, as it is difficult to locate the ablation target. PATIENT CONCERNS: 2 cases were admitted to Xiangya Hospital, Central South University with triplet pregnancies at 23 or 21weeks of gestation. DIAGNOSES: Case 1 was a 29-year-old woman with a triplet pregnancy in 2 distinct amniotic sacs and 1 fetus with multiple malformations. Case 2 was a 32-year-old woman who was identified as a triplet pregnancy with TRAP sequence with an acardiac/acephalic twin and anterior placenta. INTERVENTIONS: Both of the 2 cases were underwent a new method for MFPR involving fine needle amniotic fluid aspiration and injection of hypertonic sodium chloride (10% NaCl) into the Wharton jelly of the umbilical cord. OUTCOMES: The 2 cases resulted in selective feticide and the birth of the remaining infants from the triplet pregnancies. All infants were healthy at birth and the 2-year follow-up. LESSONS: The new approach provided a safer, more accessible, and more cost-effective method for MFPR in MMPs with a contraindication to fetoscopic surgery compared to radiofrequency ablation and fetoscopic laser surgery.

[Clinical analysis of pregnancy complicated with systemic lupus erythematosus].

OBJECTIVE: To investigate the interacting effects between pregnancy and flares of systemic lupus erythematosus (SLE) and to explore the best occasion for SLE patients' conception and the management during the pregnancy. METHODS: Thirty one cases of pregnancy complicated with SLE were investigated retrospectively, among whom 18 were in remission of SLE at the beginning of conception (Group A), and the other 13 either had high-activity of the disease or were first diagnosed as SLE during the pregnancy (Group B). Various doses of prednisone were administered to control SLE. RESULTS: SLE flares still occurred in 6 cases in Group A, but in all cases in Group B. Compared with Group A, the rates of fetal loss and early delivery were significantly higher in Group B (P < 0.05), while the survival rate and the weight of the new born were notably decreased in Group B (P < 0.05). CONCLUSION: Pregnancy and SLE interacted with each other unfavorably. Selection of remission stage for conception and proper management during the pregnancy could significantly improve the maternal-fetal safety.

[Fetal posterior cranial fossa in the second and third trimester].

OBJECTIVE: To examine the normal range of the width of posterior cranial fossa (WPCF) in the second and third trimester by ultrasonography, and to investigate its relationship with fetal congenital and chromosome abnormality. METHODS: WPCF of 2484 fetus (gestational age from 14 to 41 weeks) was measured by ultrasonograph routinely, and the infants were followed up. RESULTS: In 2848 fetus, 2772 were normal and 76 were abnormal. WPCF increased before 32 weeks, decreased after 33 weeks, the largest value of WPCF was 13.4 mm. The occurrence rate of WPCF> or =8 mm in normal fetus was 8.84%, and that in abnormal fetus was 17.46%. Most fetuses with chromosome abnormality had normal WPCF in the second trimester, but some fetuses with remarkable broadening in the late stage. Some abnormal fetuses (such as water head, Dandy-Walker's syndrome etc) showed significant extension of WPCF. CONCLUSION: WPCF increases before 32 weeks, decreases after 33 weeks;and can be easily measured during 29 - 32 weeks. WPCF of some fetus with chromosome abnormality or with congenital abnormality is remarkably broadened in the late stage. The fetus of WPCF> or =10 mm should be followed up closely, and antenatal diagnosis should be done if WPCF is more than 14 mm.

[Effects of oxytocin, misoprostol and nimodipine on expression of L-type voltage dependent calcium channel mRNA of the uterine myometrium and left ventricular myocardial cells in late pregnant rats].

OBJECTIVE: To study the effect of oxytocin, misoprostol and nimodiping on expressions of L-type voltage dependent calcium channel (VDCC-L) alpha(1) and VDCC-L beta(2) mRNA in the myometrium and left ventricular myocardial cells of the late pregnant rats. METHODS: A reverse transcription polymerase chain reaction (RT-PCR) method was used to examine and quantitate VDCC-L alpha(1) and VDCC-L beta(2) mRNA expressions in the uterine myometrium and left ventricular myocardial cells of rats, which were divided into the nature labor group, oxytocin group (oxytocin inducing labor), misoprostol group (misoprostol inducing labor) and nimodipine group (active labor at 48 hours after administering nimodipine). RESULTS: (1) In the uterine myometrium, the levels of VDCC-L alpha(1) mRNA expression in the nature labor, oxytocin, misoprostol and nimodipine group were 0.800 3 +/- 0.165 9, 0.863 1 +/- 0.192 1, 0.812 0 +/- 0.173 4 and 0.742 6 +/- 0.182 6, respectively. No significant difference was found in the four groups (P > 0.05). The levels of VDCC-L beta(2) mRNA expression in the above four groups were 0.646 9 +/- 0.130 4, 0.506 2 +/- 0.147 2, 0.500 5 +/- 0.135 6 and 0.492 9 +/- 0.127 6, respectively. There was no remarkable difference between the nature labor group and the other three administering drugs groups (P > 0.05). (2) In the left ventricular myocardial cells, expressions of VDCC-L alpha(1) in the nature labor, oxytocin, misoprostol and nimodipine group were 0.662 5 +/- 0.180 1, 0.636 5 +/- 0.157 8, 0.591 7 +/- 0.141 3 and 0.542 6 +/- 0.143 6, respectively; the levels of VDCC-L beta(2) mRNA expression were 0.670 2 +/- 0.140 5, 0.606 2 +/- 0.143 9, 0.591 4 +/- 0.121 9 and 0.585 2 +/- 0.131 0, respectively. Although both the levels of VDCC-L alpha(1) and VDCC-L beta(2) mRNA expression in the nature labor group were a little higher than those in the other three experiment groups, the statistic difference was not noted (P > 0.05). CONCLUSIONS: Oxytocin, misoprostol or nimodipine can induce or inhibit labor through regulating expressions of VDCC-L alpha(1) and VDCC-L beta(2) mRNA in the rat uterine myometrium and it may not have an adverse effect on heart function of normal pregnant rats. VDCC-L may be the common channel of labor induced by internal or external factors.

[Effects of calponin-1 gene silencing on the biological behavior of uterine smooth muscle cells].

OBJECTIVE: To study the effects of calponin-1 expression inhibition on the proliferation , invasiveness, apoptosis and cytoskeleton of uterine smooth muscle cells, and explore the molecular mechanism of calponin-1 in the uterine smooth muscle cells for labor onset. METHODS: siRNA-calponin-1 adenovirus plasmid was constructed and transfected into primarily cultured uterine smooth muscle cells. The proliferation, invasiveness and apoptosis of the cells were determined by MTT assay, matrigel invasion assays and flow cytometry, respectively. Rhodamine-Phalloidin was used for labeling filamentous actin (F-actin), and the morphology and the distribution of F-actin was observed under fluorescence microscopy and analyzed quantitatively. RESULTS: The motor ability of uterine smooth muscle cells decreased significantly after transfection with siRNA-calponin-1 adenovirus plasmid (P<0.05). The transfected cells showed thinner, loosened and irregular F-actin microfibers, and the cells in the empty vector and blank control groups showed thicker and longer F-actin microfibers. CONCLUSION: Inhibition of calponin-1 expression can inhibit uterine smooth muscle cell migration and cause the morphological change and rearrangement of F-actin without affecting its proliferation and apoptosis in vitro, suggesting that the morphological change and rearrangement of F-actin of uterine smooth muscle cell may be one of the important mechanisms in the labor onset.