Assessment of serum levels and placental bed tissue expression of IGF-1, bFGF, and PLGF in patients with placenta previa complicated with placenta accreta spectrum disorders.

OBJECTIVE: This study aims to detect the serum levels of IGF-1, bFGF, and PLGF and their expressions in placental bed tissues of patients with placenta previa complicated with PAS disorders. METHODS: This case and control study included 40 multiparous pregnant women with complete placenta previa between 34 weeks and 38 weeks of gestation and they were divided into two groups: 25 patients with PAS (case group) and 15 patients without PAS (control group). The venous blood samples were collected 2 h before the cesarean section, and the placental bed tissues were taken intraoperatively at the placental implantation site and then were histologically examined to evaluate the gravity of the myometrial invasion of the placenta. According to FIGO PAS increasing grading, the 25 patients were also divided into three groups: PAS grade I group, PAS grade II group, and PAS grade III group. The concentrations of IGF-1, bFGF, and PLGF in serum were measured using ELISA, and the mean ratio of the relative mRNA expression of each biomarker in placental bed tissues was calculated using qRT-PCR. The staining intensity and the positive cells were quantitatively measured and expressed as means by using Image J software for IHC analysis. RESULTS: IGF-1 had low serum levels and high placental bed expression in placenta previa patients with PAS disorders compared to those without PAS (all p < 0.0001). PLGF had high serum levels (p = 0.0200) and high placental bed expression (p < 0.0001) in placenta previa patients with PAS disorders compared to those without PAS. IGF-1 serum levels decreased up to PAS grade II (means were 24.3 ± 4.03, 21.98 ± 3.29, and 22.03 ± 7.31, respectively for PAS grade I, PAS grade II, PAS grade III groups, p = 0.0006). PLGF serum levels increased up to PAS grade II (means were 12.96 ± 2.74, 14.97 ± 2.56, and 14.89 ± 2.14, respectively for the three groups, p = 0.0392). However, IGF-1 and PLGF mRNA placental bed expression increased up to PAS grade III. The relative expression of mRNA means for the three groups was 3.194 ± 1.40, 3.509 ± 0.63, and 3.872 ± 0.70, respectively for IGF-1; and 2.784 ± 1.14, 2.810 ± 0.71, and 2.869 ± 0.48, respectively for PLGF (all p < 0.0001). Their IHC (immunohistochemical) staining also had increasing trends, but p > 0.05. bFGF was not significantly expressed in placenta previa with PAS disorders in most of the analysis sections (p > 0.05). CONCLUSIONS: Low serum levels and high expression in placental bed tissues of IGF-1, or high serum levels and high expression in placental bed tissues of PLGF, may differentiate placenta previa patients with FIGO PAS grade I and PAS grade II from those without PAS disorders. However, they could not significantly predict the degree of placental invasiveness in FIGO PAS grades II and III.

Placenta Accreta Spectrum Diagnosis Challenges and Controversies in Current Obstetrics: A Review.

Background: Placenta accreta spectrum (PAS) is the most common obstetric complication in current obstetrics in which the placenta is fully or partially attached to the uterine myometrial layer at delivery. This is commonly due to the deficiency of the uterine interface between the uterine endometrial and myometrial layers leading to abnormal decidualization at the uterine scar area, which permits the abnormally placental anchoring villous and trophoblasts, deeply invade the myometrium. The prevalence of PAS is globally at rising trends every day in modern obstetrics originally due to the high increasing rate of cesarean sections, placenta previa, and assisted reproductive technology (ART). Thus, the early and precise diagnosis of PAS is imperative to prevent maternal intrapartum or postpartum bleeding complications. Objective: The main aim of this review is to debate the current challenges and controversies in the routine diagnosis of PAS diseases in obstetrics. Data Source: We retrospectively reviewed the recent articles on different methods of diagnosing PAS in PubMed, Google Scholar, Web of Science, Medline, Embase, and other website databases. Results: Despite that, the standard ultrasound is a reliable and key tool for the diagnosis of PAS, the lack of ultrasound features does not exclude the diagnosis of PAS. Therefore, clinical assessment of risk factors, MRI tests, serological markers, and placental histopathological tests are also indispensable for the prediction of PAS. Previously, limited studies reached a high sensitivity rate of diagnosis PAS in appropriate cases, while many studies recommended the inclusion of different diagnosis methods to improve the diagnosis accuracy. Conclusion: A multidisciplinary squad with well-experienced obstetricians, radiologists, and histopathologists should be involved in the establishment of the early and conclusive diagnosis of PAS.

Netrin-1 promotes the vasculogenic capacity of human adipose-derived stem cells.

Adipose derived stem cells (ADSCs) have been increasingly explored for use in cell-based therapy against ischemic diseases. However, unsatisfactory angiogenesis limits the therapeutic efficacy. Netrin-1, a known axon guidance molecule, improves neovascularization in the ischemic region. Thus, our study was performed to evaluate the potential effect of Netrin-1 on the angiogenic behaviors of human ADSCs (hADSCs). hADSCs acquired from human abdominal adipose tissue were modified by liposome transfection of Netrin-1 plasmid, and the proliferation of hADSCs was determined by Cell Counting Kit-8 (CCK-8) assay. The transcript levels of pro-invasive proteins such as matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP-9), were measured to test migratory and invasive capabilities, and the levels of vascular endothelial growth factors were assayed to monitor angiogenic activity. Our results showed that Netrin-1 overexpression enhanced the proliferation of hADSCs, and promoted the migration and invasion of hADSCs, as indicated by increased levels of MMP-2 and MMP-9. Furthermore, Netrin-1 overexpression increased the expression of vascular endothelial growth factor and placental growth factor in hADSCs. Our results highlighted the possibility that genetic modification of hADSCs by Netrin-1 overexpression might be beneficial for cell transplantation therapy against ischemic diseases.

Decline of three farmland pest species in rapidly urbanizing landscapes.

Urbanization is a pressing challenge for earth's humans because it is changing not only natural environments but also agricultural lands. Yet, the consequences of cropland loss on pest insect populations that largely depend on these habitats remain largely unclear. We used a 17-year data set to investigate the dynamics of three moth pest species (i.e., striped stem borer, yellow stem borer, and pink stem borer) and their driving forces across the largest mega-urban region of China. Total abundance of three pest species is declined by about 80%, which was strongly associated with cropland loss during rapid urbanization. Our findings indicate that not only the increasing conversion of natural areas to human-dominated landscapes but also that of agricultural lands to urban landscapes can be critical to insect populations. It is therefore essential to monitor and understand the insect dynamics in rapidly urbanizing regions, which are currently found in many developing countries worldwide.

Accumulation of antibiotic resistance genes in full-scale drinking water biological activated carbon (BAC) filters during backwash cycles.

Biological activated carbon (BAC) filtration, a process widely used in drinking water treatment, was recently reported to harbor antibiotic resistance genes (ARGs). This emerging contamination is poorly understood. This study was conducted to investigate the occurrence of ARGs and bacterial community in full-scale BAC filters during the backwash cycle using high-throughput qPCR and high-throughput sequencing. A total of 178 ARGs were detected in all biofilm samples, with relative abundance ranging from 0.1 to 1.37 copies per 16S rRNA and absolute abundance ranging from 4.48 × 107 to 3.09 × 109 copies/g carbon. Biofilms sampled from different filters shared most detected ARGs and dominant genera including Bryobacter, Pedomicrobium, Reyranella, and Terrimonas, though their bacterial community structure differed significantly. After backwashing, the relative ARGs abundance increased by 1.5- to 3.8-folds and the absolute ARGs abundance increased by 0.90- to 1.12-logs in all biofilm samples during filter ripening, indicating that ARGs accumulated in filters during this period. Redundancy analysis suggested that such ARGs accumulation was mainly driven by horizontal gene transfer in winter, but highly correlated with the increasing relative abundance of genera Bryobacter and Acidibacter in summer. It was observed that 80.6 %-89.3% of the detected ARGs persisted in the filters despite of the backwashing. Given the high richness and relative abundance of ARGs in BAC filter and the ineffectiveness of backwashing in ARG removal, more stringent downstream disinfection strategies are deserved and more research is necessary to assess potential human health risks due to the persistence of ARGs in drinking water.

Detection and distribution of vbnc/viable pathogenic bacteria in full-scale drinking water treatment plants.

Viable but non-culturable (VBNC) bacteria have attracted widespread attention since they are inherently undetected by traditional culture-dependent methods. Importantly, VBNC bacteria could resuscitate under favorable conditions leading to significant public health concerns. Although the total number of viable bacteria has been theorized to be far greater than those that can be cultured, there have been no reports quantifying VBNC pathogenic bacteria in full-scale drinking water treatment plants (DWTPs). In this work, we used both culture-dependent and quantitative PCR combination with propidium monoazide (PMA) dye approaches to characterize cellular viability. Further, we established a method to quantify viable pathogens by relating specific gene copies to viable cell numbers. Ratios of culturable bacteria to viable 16S rRNA gene copies in water and biological activated carbon (BAC) biofilms were 0-4.75% and 0.04-56.24%, respectively. The VBNC E. coli, E. faecalis, P. aeruginosa, Salmonella sp., and Shigella sp. were detected at levels of 0-103 cells/100 mL in source water, 0-102 cells/100 mL in chlorinated water, and 0-103 cells/g in BAC biofilms. In addition, differences between the total and viable community structures after ozonation and chlorination were investigated. The relative abundance of opportunistic pathogens such as Mycobacterium, Sphingomonas, etc. increased in final water, likely due to their chlorine resistance. In summary, we detected significant quantities of viable/VBNC opportunistic pathogens in full-scale DWTPs, confirming that traditional, culture-dependent methods are inadequate for detecting VBNC bacteria. These findings suggest a need to develop and implement rapid, accurate methods for the detection of VBNC pathogenic bacteria in DWTPs to ensure the safety of drinking water.

Challenges for Obstetricians and the Countermeasures of COVID-19 Epidemic.

Since the outbreak of the novel coronavirus in Wuhan, China, as obstetricians, we also face great challenges. We need to identify pregnant patients with 2019 coronavirus disease infection timely, and give them appropriate treatment in order to obtain a good maternal and infant prognosis. Here, we would like to share a case and provide some suggestions on how to screen, diagnose and treat pregnant women with 2019 coronavirus disease infection during the outbreak.

Evaluation of RNA degradation in pure culture and field Microcystis samples preserved with various treatments.

RNA-based molecular technique (RT-qPCR) is a promising method for microcystin monitoring in lakes and reservoirs, but great lability of RNA in cyanobacterial samples limits its application. To date, no studies have investigated how to effectively preserve RNA in cyanobacterial samples. In this study, four different treatments (-80 °C freezer, -196 °C liquid nitrogen, 4 °C or 25 °C preservation after adding RNA protective fluid) were employed to preserve RNA in pure culture and field Microcystis samples, and RNA degradation in these treatments were systematically evaluated. Results showed liquid nitrogen was the most effective treatment to preserve RNA in pure culture and field Microcystis samples. RNA preservation using RNA protective fluid was temperature dependent. Low temperature (4 °C) could effectively slow down RNA degradation within a short time (1-7 d), since decay rate of mcyH mRNA (k = 0.00094 d-1) was much lower at 4 °C than that at 25 °C (0.0549 d-1) (P < 0.05). However, for field samples, RNA degradation was much faster than pure culture samples with the same treatment. Therefore, to better preserve RNA in field samples, a practical strategy for RNA preservation combining RNA protective fluid and liquid nitrogen, was proposed. Tests of field experiments showed it was more effective than individual treatment for RNA preservation in Microcystis samples during field sampling. Thus, this strategy could be employed to preserve RNA in cyanobacterial samples during field sampling, which will contribute to the application of RT-qPCR technique for microcystin monitoring in lakes and reservoirs.

Efficient One-Step Induction of Human Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) Produces MSC-Derived Neurospheres (MSC-NS) with Unique Transcriptional Profile and Enhanced Neurogenic and Angiogenic Secretomes.

Cell therapy has emerged as a promising strategy for treating neurological diseases such as stroke, spinal cord injury, and various neurodegenerative diseases, but both embryonic neural stem cells and human induced Pluripotent Stem Cell- (iPSC-) derived neural stem cells have major limitations which restrict their broad use in these diseases. We want to find a one-step induction method to transdifferentiate the more easily accessible Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) into neural stem/progenitor cells suitable for cell therapy purposes. In this study, UC-MSCs were induced to form neurospheres under a serum-free suspension culture with Epidermal Growth Factor- (EGF-) and basic Fibroblast Growth Factor- (bFGF-) containing medium within 12 hours. These MSC-derived neurospheres can self-renew to form secondary neurospheres and can be readily induced to become neurons and glial cells. Real-time PCR showed significantly upregulated expression of multiple stemness and neurogenic genes after induction. RNA transcriptional profiling study showed that UC-MSC-derived neurospheres had a unique transcriptional profile of their own, with features of both UC-MSCs and neural stem cells. RayBio human growth factor cytokine array analysis showed significantly upregulated expression levels of multiple neurogenic and angiogenic growth factors, skewing toward a neural stem cell phenotype. Thus, we believe that these UC-MSC-derived neurospheres have amenable features of both MSCs and neural stem/progenitor cells and have great potential in future stem cell transplantation clinical trials targeting neurological disorders.

EMT induced by loss of LKB1 promotes migration and invasion of liver cancer cells through ZEB1-induced YAP signaling.

Liver cancer cells often exhibit mesenchymal phenotypes, a critical phenotypic alteration of cancer cells termed the epithelial-mesenchymal transition (EMT). To examine whether liver kinase B1 (LKB1) serves a potential role in EMT in liver carcinogenesis, in the present study, it was determined that the expression of LKB1 decreased in the hepatocellular carcinoma (HCC) cell line, compared with a normal liver cell line. LKB1 overexpression decreased cell motility and invasiveness. Furthermore, the loss of LKB1 induced the expression of several EMT marker proteins, including that of Zinc Finger E-Box Binding Homeobox 1 (ZEB1). Notably, the expression of Yes-associated protein (YAP) was positively associated with that of ZEB1 in LKB1-knockdown cells with a mesenchymal phenotype. Here, we describe the direct regulation of the Hippo pathway effector YAP by ZEB1. The findings of the present study demonstrate that ZEB1 regulates the expression of YAP and regulates the expression of downstream target genes to promote malignant progression.

Conditioned Medium from Human Umbilical Vein Endothelial Cells Promotes Proliferation, Migration, Invasion and Angiogenesis of Adipose Derived Stem Cells.

Preeclampsia (PE) is a pregnancy-specific hypertensive complication, closely related to endothelial dysfunction. Adipose derived stem cells (ADSCs) have the capacity to differentiate into endothelial cells for vascular repair. Therefore, we hypothesized that induced endothelial differentiation of ADSCs might hold great potential for the treatment of PE. In this study, the primary ADSCs and human umbilical vein endothelial cells (HUVECs) were isolated by the collagenase digestion method. The supernatant of HUVECs was collected from the first generation of cells. Then, ADSCs were divided into two groups: ADSCs alone group and induced ADSCs (iADSCs) group. In iADSCs group, ADSCs were induced by HUVECs conditioned medium and ADSCs special culture medium at a ratio of 1:1 over a two-week period. In order to identify the endothelial characteristics of iADSCs, CD31 and CD34 were examined by flow cytometry. The proliferation, migration, invasion and angiogenesis assays were employed to compare the bioactivity of iADSCs and ADSCs. Furthermore, The levels of angiogenic related factors including vascular endothelial growth factor (VEGF) and placenta growth factor (P1GF) were detected by RT-PCR and Western blotting. Results showed conditioned medium from HUVECs promoted ADSCs proliferation, migration, invasion and angiogenesis. In addition, the levels of VEGF and P1GF were significantly enhanced in iADSCs group. This study uncovered the iADSCs application potential in the therapy and intervention of PE.

Transplantation of endothelial progenitor cells for improving placental perfusion in preeclamptic rats.

OBJECTIVE: Effective treatments for preeclampsia are currently unavailable. As endothelial progenitor cell-transplantation may improve ischemia, it is an important undertaking to study the role of endothelial progenitor cells for improving the symptom of preeclampsia. METHOD: Physiological and pathological changes in foetal rats and pregnant rats were monitored. Endothelial progenitor cells were isolated from the peripheral blood of normal rats and labelled by DiI. Endothelial progenitor cells were transplanted into the placenta of preeclampsia-like rats. Fluorescence microscopy was used to observe the differentiation of transplanted endothelial progenitor cells. Western blotting was used to observe the expression of nestin, an index of brain hypoxia in foetal rats. RESULT: The rats suffered from abdominal aortic constriction and NG-nitro-L-arginine methyl ester injection (group F). The proteinuria and blood pressure of pregnant rats in group F increased on the 13th day of pregnancy. The proteinuria and blood pressure of group F was higher than in other groups of rats. The weight of foetal rats and foetal heads significantly decreased in group F compared with other groups. Typical pathological changes of preeclampsia were observed in the placental tissue of group F. In preeclampsia-like rats, transplantation of endothelial progenitor cells led to an increase in placenta angiogenesis. The expression of nestin weakened in endothelial progenitor cell-transplanted rats compared with the non-transplantation group. After EPCs transplantation, physiological parameters in the preeclampsia-like rats were significantly decreased. CONCLUSION: Endothelial progenitor cells transplantation could improve preeclampsia-like symptom in rats and endothelial progenitor cell-transplantation relieves intrauterine hypoxia in brain tissues of foetal rats to a certain extent.

Simple and effective large-scale preparation of geniposide from fruit of Gardenia jasminoides Ellis using a liquid-liquid two-phase extraction.

Geniposide was prepared on a large-scale using a selective two-phase liquid-liquid extraction. The aqueous residue from the fruit of Gardenia jasminoides Ellis was treated with sodium carbonate and extracted with n-butanol several times. The n-butanol extracts were treated with activated granular charcoal to remove pigments and were then concentrated to produce a residue with a high solid content. The residue was crystallized to obtain geniposide with 98% purity. For large-scale synthesis, the residue (solid content 45%, geniposide 5.5%) was extracted to generate 70g of geniposide with 98% purity and 84.8% recovery using 1500g residue.

[In vivo and in vitro research study of the influence on placenta angiogenesis by gene silencing of netrin-1].

OBJECTIVE: To study influence on angiogenesis of placenta by gene silencing of netrin-1. METHODS: Netrin-1 gene in human umbilical vein endothelial cells (HUVEC) and placenta of pregnant rats were silenced by RNA interference. The following methods were used in this study, including the phenytetrazoliumromide (MTT) for viability, clone formation for proliferation, transwell for migration, and tube formation for angiogenesis in vitro. The change of fetal growth was recorded. Placental microvessel density in pregnant rats was measured by immunohistochemical CD(34) staining in vivo. RESULTS: (1) HUVEC: viability and proliferation of HUVEC were remarkably inhibited by gene silencing of netrin-1, which number of clone formation, migration cell, tube formation were from (69 ± 6)%, 86 ± 17, 37 ± 9 decreased to (46 ± 5)%, 46 ± 13 and 17 ± 5 (P < 0.05) respectively. (2) Placenta of pregnant rats: after netrin-1 gene silenced, fetal weight were decreased from (2.39 ± 0.17) g to (2.12 ± 0.10) g (P < 0.05). Placental microvessel density was decreased from (258 ± 38)/mm(2) to (197 ± 32)/mm(2) in vivo (P < 0.05). CONCLUSIONS: Gene silencing of netrin-1 could inhibit viability, proliferation, migration, tubal formation of HUVEC and angiogenesis of placenta. Netrin-1 plays an important role in regulating angiogenesis in placenta.

Role of axonal guidance factor netrin-1 in human placental vascular growth.

This study investigated the role of netrin-1 in placental vascular development. In vitro rat aortic ring assay and in vivo Matrigel plug assay were conducted to exmaine the effect of netrin-1 on angiogenesis. Human placental microvascular endothelial cells (HPMECs) were isolated and cultured and their viability, migration and tubular formation were studied, in order to examine the effects of netrin-1. The results showed that netrin-1 potently stimulated neovascularization in a mouse Matrigel plug in vivo and the sprouting of endothelial cells in rat aortic rings in vitro. In addition, netrin-1 enhanced the viability, migration and tube formation of HPMECs. Our study suggested that netrin-1 could significantly promote the formation of blood vessels of human placenta and may be a potential target for developing new therapeutic strategies for placental vasculature-related diseases.

Evaluation of "J"-shaped uterine incision during caesarean section in patients with placenta previa: a retrospective study.

This study evaluated the efficacy and safety of "J"-shaped uterine incision for caesarean section for patients diagnosed with placenta previa. A total of 55 consecutive cases of placenta previa treated in Union Hospital were retrospectively analyzed over a period of two years and 10 months. The subjects were divided into two groups with respect to the uterine incision. Twenty-four pregnant women with placenta previa who were indicated for caesarean section underwent the procedure using a new "J"-shaped uterine incision and 31 pregnant women with placenta previa received caesarean section that used the traditional transverse incision. The two groups were compared in terms of operation time, estimated blood loss, infant expulsion time, exhaust time and postoperative recovery. Meanwhile, comparison was also made in neonatal clinical data between the two groups. Compared with the "J"-shaped incision group, the traditional incision group had a lower Apgar scores (P<0.05). However, there existed no statistically significant differences in the overall time of operation and postoperative period of breaking wind (P>0.05). It is concluded that, with caesarean section for placenta previa patients, the "J"-shaped uterine incision significantly decreases intraoperative blood loss and facilitates the fetal delivery.

[Correlation of netrin-1 expression with invasion of extra villous trophoblasts].

OBJECTIVE: To investigate mechanism of netrin-1 regulating invasion of extra villous trophoblasts. METHODS: RT-PCR was used to detect six receptors expression including UNC5A, UNC5B, UNC5C, UNC5D, DCC and neogenin in extra villous trophoblast cell line TEV-1. The TEV-1 cells were cultured and devided into seven groups according to the concentration of netrin-1 adding into the medium, which include 10 microg/L, 50 microg/L, 100 microg/L, 500 microg/L, 1000 microg/L, 5000 microg/L and the control (the concentration of netrin-1 was 0 microg/L)groups. The proliferation and invasion of TEV-1 induced by netrin-1 were determined by CCK-8 assay and transwell invasion assay respectively. RESULTS: (1) Only neogenin and UNC5B were found to be expressed on TEV-1 by RT-PCR method. (2) In CCK-8 proliferation assay, after 72 hours culture, the proliferation of TEV-1 were 1.55 +/- 0.29 in 10 microg/L, 1.72 +/- 0.31 in 50 microg/L, 2.15 +/- 0.35 in 100 microg/L, 1.42 +/- 0.25 in 500 microg/L, 1.50 +/- 0.27 in 1000 microg/L, and 1.38 +/- 0.23 in 5000 microg/L group, which were all higher than 1.00 +/- 0.16 in control group significantly (P < 0.05). (3) In matrigel invasion assay, after 6 hours culture, the number of the trans-membrane cells in various netrin-1 group, including 41 +/- 4 in 10 microg/L, 47 +/- 5 in 50 microg/L, 55 +/- 6 in 100 microg/L, 44 +/- 5 in 500 microg/L, 43 +/- 5 in 1000 microg/L and 42 +/- 5 in 5000 microg/L group, were all higher than 30 +/- 4 in control group with statistical significance (P < 0.05). (4) The fold changes of neogenin were 1.50 +/- 0.16 in 10 microg/L, 1.83 +/- 0.19 in 50 microg/L, 2.24 +/- 0.25 in 100 microg/L, 2.12 +/- 0.24 in 500 microg/L, 2.12 +/- 0.23 in 1000 microg/L and 2.13 +/- 0.23 in 5000 microg/L group, which were all higher than 1.00 +/- 0.11 in control group significantly (P < 0.05). There were significant difference between group 10 microg/L and 50 microg/L, group 50 microg/L and 100 microg/L (P < 0.05). There were no significant difference between group 100 microg/L and 500 microg/L, group 1000 microg/L and 5000 microg/L (P > 0.05). (5) The fold changes of UNC5B 1.09 +/- 0.11 in 10 microg/L, 1.47 +/- 0.14 in 50 microg/L, 1.61 +/- 0.16 in 100 microg/L, 1.85 +/- 0.19 in 500 microg/L, 2.21 +/- 0.21 in 1000 microg/L and 2.42 +/- 0.23 in 5000 microg/L group, were all higher significantly when compared with 1.00 +/- 0.07 in control group (P < 0.05). There were significant difference between all groups (P < 0.05). CONCLUSION: Netrin-1 can promote the potential of proliferation and invasion of extravillous trophoblasts in vitro through its receptors including neogenin and UNC5B.

Changes in number and biological function of endothelial progenitor cells in hypertension disorder complicating pregnancy.

To examine the changes in number and function of endothelial progenitor cells (EPCs) from peripheral blood (PB) in hypertension disorder complicating pregnancy (HDCP), 20 women with HDCP and 20 normal pregnant women at the third trimester were studied. Mononuclear cells (MNCs) from PB were isolated by Ficoll density gradient centrifugation. EPCs were identified by positive expression of both CD34 and CD133 under fluorescence microscope and positive expression of factor VIII as shown by immunocytochemistry. The number of EPCs was flow-cytometrically determined. Proliferation and migration of EPCs were measured by MTT assay and modified Boyden chamber assay, respectively. The adhesion activity of EPCs was detected by counting the number of the adherent cells. The results showed that, compared with normal pregnant women, the number of EPCs was significantly reduced in HDCP (4.29%+/-1.21% vs 15.32%+/-2.00%, P<0.01), the functional activity of EPCs in HDCP, such as proliferation (13.45%+/-1.68% vs 18.45%+/-1.67%), migration (37.25+/-7.28 cells/field vs 67.10+/-9.55 cells/field) and adhesion activity (20.65+/-5.19 cells/field vs 34.40+/-6.72 cells/filed) was impaired (P<0.01). It is concluded that the number and function of EPCs are significantly decreased in HDCP.

Effects of Smac gene over-expression on the radiotherapeutic sensitivities of cervical cancer cell line HeLa.

BACKGROUND: The second mitochondria-derived activator of caspases (Smac) is a novel proapoptotic gene, which plays an important role in the apoptosis-inducing effects of irradiation on tumor cells. The purpose of this study was to investigate the effects of extrinsic Smac gene transfer and its over-expression in radiotherapeutic sensitivities of cervical cancer cells. METHODS: After the Smac gene was transferred into the cervical cancer cell line HeLa, subcloned cells were obtained by persistent G418 selection. Cellular Smac gene expression was detected by RT-PCR and Western blot, while in vitro cell viabilities were detected by trypan blue staining assay. After treatment with X-ray irradiation, cellular radiotherapeutic sensitivities were investigated by tetrazolium bromide colorimetry. Cellular apoptosis and its rate were determined by electronic microscopy, annexin V-FITC and propidium iodide staining flow cytometry. The expression and activities of cellular caspase-3 were assayed by Western blot and colorimetry. RESULTS: Smac mRNA and protein levels in HeLa/Smac cells and the selected subclone cell line of cervical cancer were significantly higher than those of HeLa (P < 0.01). There was no significant difference in cellular viabilities between them (P > 0.05). However, after irradiation with 8 Gy X-ray, growth activities of HeLa/Smac were reduced by 22.42% (P < 0.01). When compared with those of HeLa, partial HeLa/Smac cells presented characteristic morphological changes of apoptosis under electronic microscope, with higher apoptosis rates (16.4% vs. 6.2%, P < 0.01); the caspase-3 expression levels in HeLa/Smac cells were improved significantly (P < 0.01), while its activities were increased by 3.42 times (P < 0.01). CONCLUSIONS: Stable transfer of the extrinsic Smac gene and its over-expression in cervical cancer cell line could significantly enhance the expression and activities of cellular caspase-3 and ameliorate apoptosis-inducing effects of irradiation on cancer cells, which was a novel strategy to improve radiotherapeutic effects on cervical cancer.

The value of the soluable intercellular adhesion molecule-1 levels in matermal serum for determination of occult chorioamnionitis in premature rupture of membranes.

To compare the diagnostic value of soluble intercellular adhesion molecule 1 (sICAM-1) with that of c-reactive protein (CRP) for detecting chorioamnionitis (CAM) in serum of women with premature rupture of membranes (PROM), 55 pregnant women with PROM, including 18 pregnant women with preterm premature rupture of membranes (PPROM) and 20 normal pregnant women at term (TPROM) were studied. Maternal serum were measured by Sandwish enzyme-linked immunoabsorbent assay (ELISA) for sICAM. CAM was histologically confirmed after delivery. The results revealed that (1) maternal serum levels of sICAM-1 and CRP were significantly higher in women with PROM than those without it; (2) maternal serum levels of sICAM-1 and CRP were significantly higher in women with CAM than those without it; (3) serum levels of sICAM-1 in PPROM women were similar to those in TPROM women, whereas serum levels of CRP in PPROM women were significantly higher than those in TPROM women; (4) the sensitivity, specificity, positive predictive value, negative predictive value, Kappa index and area under receiver operating characteristic (ROC) curve of maternal serum sICAM-1 (cutoff 104.7 ng/ml) and CRP (cutoff 1.03 mg/dl) for diagnosing CAM were 100%, 91.2%, 87.5%, 100%, 0.20, 0.995 and 81.0%, 73.5%, 65.4%, 86.2%, 0.13, 0.811, respectively; (5) among the mild histological CAM group, severe histological CAM group and clinical CAM group, the difference in maternal serum levels of sICAM-1 were significantly (P<0.001), with the order of concentration from high level to low level corresponding to the severity of CAM. It is concluded that maternal serum level of ICAM-1 is superior to that of CRP as biomarker for diagnosing intraamniotic infection in pregnant women with PROM.