DNA-Peptide Interaction-Modulated Charge Reversal in Biomimetic Nanochannels for Simple and Efficient Detection of Histone Deacetylases.

Protein acetylation, a fundamental post-translational modification, plays a critical role in the regulation of gene expression and cellular processes. Monitoring histone deacetylases (HDACs) is important for understanding epigenetic dynamics and advancing the early diagnosis of malignancies. Here, we leverage the dynamic characteristics of DNA-peptide interactions in biomimetic nanochannels to develop a HDAC detection method. In specific, the catalysis of peptide deacetylation by HDACs triggers alterations in the charge states of the nanochannel surface to accommodate DNA molecules. Then, the interaction between DNA and peptides shifts the nanochannel surface charge from positive to negative, leading to a reversal of the ion current rectification (ICR). By calculation of the ICR ratio, quantitative detection of HDACs can be efficiently achieved using the nanochannel-based method in an enzyme-free and label-free manner. Our experimental results demonstrate that HDACs can be detected by using this method within a concentration range of 0.5-500 nM. The innate simplicity and efficiency of this strategy may render it a valuable tool for advancing both fundamental research and clinical applications in the realm of epigenetics and personalized medicine.

Engineered Escherichia coli as a Controlled-Release Biocarrier for Electrochemical Immunoassay.

Micro/nanocarriers hold great potential in bioanalysis for molecular recognition and signal amplification but are frequently hampered by harsh synthesis conditions and time-consuming labeling processes. Herein, we demonstrate that Escherichia coli (Ec) can be engineered as an efficient biocarrier for electrochemical immunoassay, which can load ultrahigh amounts of redox indicators and simultaneously be decorated with detection antibodies via a facile polydopamine (PDA)-mediated coating approach. Compared with conventional carrier materials, the entire preparation of the Ec biocarrier is simple, highly sustainable, and reproducible. Moreover, immune recognition and electrochemical transduction are performed independently, which eliminates the accumulation of biological interference on the electrode and simplifies electrode fabrication. Using human epidermal growth factor receptor 2 (HER2) as the model target, the proposed immunosensor exhibits excellent analytical performance with a low detection limit of 35 pg/mL. The successful design and deployment of Ec biocarrier may provide new guidance for developing biohybrids in biosensing applications.

IGF2BP2 enhances LincRNA01116 stability via m6 A: A potential biomarker and therapeutic target for patients with pre-eclampsia.

Pre-eclampsia (PE) is a serious complication in pregnant women characterized by failure of placental remodeling and is one of the primary causes of changes in the placental structure and function. The aberrant expression of long noncoding RNA is associated with the occurrence and progression of PE. This study found that linc01116 expression was significantly downregulated in PE patients and was related to poor uterine spiral artery remodeling. Knockdown of linc01116 remarkably decreased the angiogenesis of trophoblast cells in vitro and in vivo. Mechanistically, IGF2BP2 regulated linc01116 RNA stability via m6 A methylation. Bioinformatics and other experiments further revealed that linc01116 upregulates AAMP expression by adsorbing miR-210-3p in trophoblast cells. In conclusion, this study revealed the critical role of linc01116 in regulating trophoblast angiogenesis. Furthermore, the study provides new clues for detecting placental pathology in PE.

Preparation of Well-Constructed and Metal-Modified Covalent Organic Framework Nanoparticles for Biosensing Design with Cascade Catalytic Capability.

Uniform covalent organic framework nanoparticles (COF NPs) with a well-defined pore structure may provide a robust platform for scaffolding enzymes. Herein, bipyridine-based spherical COF NPs have been successfully prepared in this work through the Schiff base condensation reaction. Moreover, they are functionalized by metal modification and are further used for biosensor fabrication. Experimental results reveal that the metal-modified COF NPs also display impressive peroxidase-like catalytic activities, while they can load enzymes, such as glucose oxidase (GOx) and sarcosine oxidase (SOx), to develop a cascade catalysis system for design of various kinds of biosensors with very well performance. For example, the optimized GOx@Fe-COFs can achieve a sensitive detection of glucose with a low limit of detection (LOD) of 12.8 µM. Meanwhile, the enzymes also exhibit a commendable preservation of 80% enzymatic activity over a span of 14 days under ambient conditions. This work may pave the way for advancing cascade catalysis and the analysis of different kinds of biological molecules based on COF NPs.

Downregulated DUXAP8 lncRNA impedes trophoblast cell proliferation and migration by epigenetically upregulating TFPI2 expression.

BACKGROUND: Preeclampsia (PE), a pregnancy complication characterized by new-onset hypertension and proteinuria during the second trimester, is the leading cause of neonatal and maternal morbidity and mortality. In the etiology of PE, failure of uterine spiral artery remodeling may be related to functioning abnormally of trophoblast cells, leading to the occurrence and progression of PE. Recently, long noncoding RNAs (lncRNAs) have been reported to play critical roles in PE nowadays. This study aimed to investigate the expression and functions of the TFPI2 pathway-related lncRNA DUXAP8. METHODS: DUXAP8 expression in the placenta from pregnancies was examined using qPCR. Then, the in vitro functions of DUXAP8 were investigated through MTT, EdU, colony, transwell, and flow cytometry experiments. The downstream gene expression profiles were assessed using RNA transcriptome sequencing analysis and verified using qPCR and western blot. Furthermore, Immunoprecipitation (RIP), chromatin immunoprecipitation (CHIP) and fluorescence in situ hybridization (FISH) were used to detect the interaction between lncDUXAP8/EZH2/TFPI2. RESULTS: The expression of lncRNA DUXAP8 in placenta of patients with eclampsia was significantly decreased. After knockout of DUXAP8, the proliferation and migration of trophoblasts were significantly decreased, and the percentage of apoptosis was increased. Flow cytometry showed that low expression of DUXAP8 increased the accumulation of cells in G2/M phase, while overexpression of DUXAP8 had the opposite effect. We also proved that DUXAP8 epigenetically inhibited TFPI2 expression by recruiting EZH2 and mediating H3K27me3 modification. CONCLUSION: Together, these resulting data clarify that aberrant expression of DUXAP8 is involved in the potential PE development and progress. Unraveling the role of DUXAP8 will provide novel insights into the pathogenesis of PE.

A novel regulatory mechanism network mediated by lncRNA TUG1 that induces the impairment of spiral artery remodeling in preeclampsia.

Preeclampsia (PE) is associated with maternal and fetal perinatal morbidity and mortality, which brings tremendous suffering and imposes an economic burden worldwide. The failure of uterine spiral artery remodeling may be related to the abnormal function of trophoblasts and lead to the occurrence and progression of PE. Aberrant expression of long non-coding RNAs (lncRNAs) is involved in the failure of uterine spiral artery remodeling. However, the regulation of lncRNA expression in PE is poorly characterized. Here, we reported that hypoxia-induced microRNA (miR)-218 inhibited the expression of lncRNA TUG1 by targeting FOXP1. Further RNA sequencing and mechanism analysis revealed that silencing of TUG1 increased the expression of DNA demethylase TET3 and proliferation-related DUSP family, including DUSP2, DUSP4, and DUSP5, via binding to SUV39H1 in the nucleus. Moreover, TUG1 modulated the DUSP family in vitro through a TET3-mediated epigenetic mechanism. Taken together, our results unmask a new regulatory network mediated by TUG1 as an essential determinant of the pathogenesis of PE, which regulates cell growth and possibly the occurrence and development of other diseases.

Fear of childbirth and its determinants in pregnant women in the third trimester: a cross-sectional study.

BACKGROUND: Fear of childbirth (FOC) is a prevalent issue among pregnant women and significantly relates to adverse outcomes for the mother and child. However, it is not clear the prevalence and risk factors of FOC among pregnant women in a region with a moderate level of economic development in China. The aim of this study was to investigate the prevalence and risk factors of FOC among pregnant women in the third trimester of pregnancy in Lianyungang city, Eastern China. METHODS: A cross-sectional survey was conducted from December 2022 to February 2023 among pregnant women in the third trimester who met the inclusion criteria and visited Lianyungang Maternal and Child Health Hospital in Jiangsu Province, Eastern China. A structured questionnaire including sociodemographic characteristics, clinical characteristics, FOC, family function, doctor-patient communication, social support, general self-efficacy, anxiety, depression, insomnia symptoms, and quality of life was used to collect data. A multiple linear regression model was used to identify predictors of FOC. RESULTS: This study included 535 pregnant women in the third trimester. The mean score of FOC was 30.67 ± 10.18, and the median score was 29.00. The prevalence of FOC was 56.64%. Multiple linear regression analysis revealed that pregnant women with electronic screen exposure time more than 5 h per day (ß = 2.02, 95%CI: 0.50-3.53, P < 0.05), no history of cesarean section (ß = 2.66, 95%CI: 0.61-4.71, P < 0.05), likes sour food or hates greasy food (ß = 1.75, 95%CI: 0.00-3.50, P < 0.05), anxiety (ß = 0.50, 95%CI: 0.21-0.80, P < 0.05) and depression (ß = 0.30, 95%CI: 0.04-0.57, P < 0.05) were more likely to have a greater level of FOC than their counterparts. However, a significantly lower level of FOC was observed in pregnant women who were multipara (ß=-1.64, 95%CI: -3.27-0.01, P < 0.05), not worrying about delivery without family members (ß=-3.75, 95%CI: -5.26-2.25, P < 0.001), had good family function (ß=-0.32, 95%CI: -0.64-0.00, P < 0.05) and doctor-patient communication (ß=-0.33, 95%CI: -0.64-0.02, P < 0.05). CONCLUSIONS: The prevalence of FOC was high in Lianyungang city, Eastern China. FOC is influenced by multiple factors. There is an urgent need to develop interventions to reduce the prevalence of FOC in the third trimester of pregnancy, and to pay attention to pregnant women with risk factors for FOC.

Prevalence and associated factors of insomnia symptoms among pregnant women in the third trimester in a moderately developing region of China.

BACKGROUND: Insomnia is the most common sleep disorder in the general population, especially among pregnant women, and it is considered a major public health issue. Not only can it cause mental and physical problems in pregnant women, but it may also affect the growth of the fetus. However, there are few reports on the prevalence and influencing factors of insomnia symptoms in third-trimester women in China. The objective of this study was to assess the prevalence of insomnia symptoms among pregnant women in the third trimester in a moderately developing region of China and to further explore the associated factors of insomnia symptoms from various aspects. METHODS: A cross-sectional survey was conducted among eligible pregnant women in the third trimester from December 2022 to February 2023. Data on socio-demographic characteristics, clinical characteristics, and behavioral and psychological characteristics of pregnant women were collected through a structured questionnaire. The Chi-square test and multivariate logistics regression were applied to explore the associated factors of insomnia symptoms. RESULTS: A total of 535 pregnant women in the third trimester were included in this study, and the prevalence of insomnia symptoms was 59.8%. Multivariate logistic regression analysis revealed that pregnant women who lived together with elders (OR: 0.58, 95% CI: 0.40-0.86), had low perceived stress (OR: 0.58, 95% CI: 0.35-0.97), had no threatened abortion (OR: 0.55, 95% CI: 0.32-0.93) and had good doctor-patient communication (OR: 0.66, 95% CI: 0.45-0.98) were more likely to stay away from insomnia symptoms. However, pregnant women with anxiety symptoms (OR: 2.27, 95% CI: 1.28-4.03), fear of childbirth (OR: 1.63, 95% CI: 1.11-2.40) and a high experience of COVID-19 fear (OR: 1.61, 95% CI: 1.03-2.54) tended to have insomnia symptoms. CONCLUSIONS: The prevalence of insomnia symptoms in pregnant women is high in Lianyungang city in eastern China in the third trimester. Insomnia symptoms is influenced by multiple factors. There is an urgent need to develop interventions to reduce the prevalence of insomnia symptoms in the third trimester and to focus on pregnant women with risk factors for insomnia symptoms.

Functionalization of Covalent Organic Frameworks with Peptides by Polymer-Assisted Surface Modification and the Application for Protein Detection.

Although covalent organic frameworks (COFs) have received extensive attention for biomedical research due to their unique properties, their application is still hindered by the challenges of incorporating COFs with functional biomolecules. Since peptides have shown advantages in biomedical applications, herein, we propose the functionalization of COFs with peptides by a polymer-assisted surface modification strategy. Furthermore, a method based on the peptide-functionalized COFs for protein detection has also been developed to demonstrate their application potential. With the help of the polymers, peptides and horseradish peroxidase are attached onto COFs with a high surface density, and the developed method has achieved simple and sensitive detection of the secreted protein acidic and rich in cysteine. We speculate that the facile method proposed in this work to prepare peptide-functionalized COFs can not only benefit protein detection but also promote more biomedical applications of COFs.

Down-regulated lncRNA AGAP2-AS1 contributes to pre-eclampsia as a competing endogenous RNA for JDP2 by impairing trophoblastic phenotype.

Recently, growing evidence has shown that aberrant long non-coding RNA (lncRNA) expression in conjunction with an impaired trophoblastic phenotype could implicate the pathological process of pre-eclampsia (PE). However, only a small portion of lncRNAs has been characterized with regard to the function and molecular mechanisms involved in PE. There are still gaps in the available knowledge; as a result, there are currently only a few applicable treatments for PE in the context of lncRNA. Here, we found that lncRNA AGAP2-AS1 is abnormally down-regulated in severe PE placenta tissues. Using human trophoblasts, we established that AGAP2-AS1 knockdown could inhibit trophoblasts proliferation and invasion and promote cell apoptosis. Further, we showed that overexpression of AGAP2-AS1 substantially stimulated the development of the trophoblastic phenotype. Through high-throughput sequencing analysis, we demonstrated that silencing of AGAP2-AS1 favourably regulated various genes which are relevant to trophoblastic growth and invasion. Mechanistically, AGAP2-AS1 promoted the suppressor protein, Jun dimerization protein 2 (JDP2), by sponging miR-574-5p. Resultantly, further impairment of the trophoblastic phenotype was achieved by way of inhibiting cell growth, apoptosis and invasion. We also determined that the expression of AGAP2-AS1 could be mediated by FOXP1. Our results showed that the down-regulated expression of lncRNA AGAP2-AS1 might serve as a key suppressor in PE via inhibition of JDP2 at the post-transcriptional level by competing for miR-574; thus, this presents a novel therapeutic strategy for PE.

Downregulation of hsa_circ_0005243 induces trophoblast cell dysfunction and inflammation via the ß-catenin and NF-κB pathways.

BACKGROUND: Gestational diabetes mellitus (GDM) is a common complication in pregnancy that poses a serious threat to the health of both mother and child. While the specific etiology and pathogenesis of this disease are not fully understood, it is thought to arise due to a combination of insulin resistance, inflammation, and genetic factors. Circular RNAs (circRNAs) are a special kind of non-coding RNA that have attracted significant attention in recent years due to their diverse activities, including a potential regulatory role in pregnancy-related diseases, such as GDM. METHODS: We previously reported the existence of a novel circRNA, hsa_circ_0005243, which was identified by RNA sequencing. In this study, we examined its expression in 20 pregnant women with GDM and 20 normal pregnant controls using quantitative reverse transcription PCR analysis. Subsequent in vitro experiments were conducted following hsa_circ_0005243 knockdown in HTR-8/SVneo cells to examine the role of hsa_circ_0005243 in cell proliferation and migration, as well as the secretion of inflammatory factors such as tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6). Finally, we examined the expression of ß-catenin and nuclear factor kappa-B (NF-κB) signaling pathways to assess their role in GDM pathogenesis. RESULTS: Expression of hsa_circ_0005243 was significantly reduced in both the placenta and plasma of GDM patients. Knockdown of hsa_circ_0005243 in trophoblast cells significantly suppressed cell proliferation and migration ability. In addition, increased secretion of inflammatory factors (TNF-α and IL-6) was observed after hsa_circ_0005243 depletion. Further analyses showed that knockdown of hsa_circ_0005243 reduced the expression of ß-catenin and increased nuclear NF-κB p65 nuclear translocation. CONCLUSIONS: Downregulation of hsa_circ_0005243 may be associated with the pathogenesis of GDM via the regulation of ß-catenin and NF-κB signal pathways, suggesting a new potential therapeutic target for GDM.

[Relationship of group B streptococcus colonization in late pregnancy with perinatal outcomes].

OBJECTIVE: To investigate the relationship of group B streptococcus (GBS) colonization in late pregnancy with perinatal outcome. METHODS: Pregnant women who underwent antenatal check-up at General Hospital of PLA Eastern Theater Command and the First Affiliated Hospital of Nanjing Medical University from January 2016 to December 2018 were enrolled in the study. The vaginal and rectal swab samples were collected for GBS culture at 35-37 weeks of pregnancy. The perinatal outcomes of positive and negative GBS groups were compared. The GBS-positive group samples were tested for antibiotic susceptibility. In GBS positive group the maternal and child perinatal outcomes were compared between pregnant women with antibiotics treatment and those without antibiotics. RESULTS: A total of 13 000 pregnant women were enrolled, and the overall colonization rate of GBS was 3.65%(475/13 000). The colonization rate of GBS in the vagina was 2.33%(303/13 000), and the colonization rate in the rectum was 1.75%(227/13 000). Through the collection and detection of rectal specimens, the positive rate of GBS increased by 56.77%(172/303). The monthly colonization rate of GBS showed significant fluctuations with the highest in March and October (all P < 0.05). The sensitivity of 475 GBS-positive specimens to ceftriaxone, vancomycin and linezolid were 100%, and the sensitivity to ampicillin and penicillin were 97.26%and 93.47%, respectively. The resistance rates of the strains to levofloxacin, clindamycin, erythromycin and tetracycline were 30.11%, 48.00%, 52.21%and 88.63%. The incidence of premature rupture of membranes, postpartum hemorrhage, puerperal infection, neonatal pneumonia and sepsis in GBS positive group were significantly higher than those in GBS negative group (all P < 0.01). In pregnant women with positive GBS, the incidence of puerperal infection, neonatal infection and admission to the NICU in the antibiotic group were significantly lower than those in the non-antibiotic group ( P < 0.05 or P < 0.01). CONCLUSIONS: The total colonization rate of GBS is low. The detection of GBS can be significantly improved by supplementing rectal examination. Ceftriaxone, ampicillin and penicillin are currently the drugs of choice for the prevention and treatment of GBS-related diseases. GBS infection can increase the incidence of maternal and child complications. The use of antibiotics during labor can improve the outcome of mothers and infants.

Resveratrol promotes trophoblast invasion in pre-eclampsia by inducing epithelial-mesenchymal transition.

Impairment spiral arteries remodelling was considered to be the underlying cause of pathogenesis of pre-eclampsia (PE). Resveratrol (RE) was reported that it could modulate cellar phenotype to ameliorate diverse human diseases. However, the biological function of RE in PE remains poorly understood. In this report, we investigated the effect of RE on trophoblast phenotype both in vivo and in vitro. We conducted MTT and transwell assays to explore cell proliferation and invasion events in HTR-8/SVneo. In mice model, the clinical characteristics of PE were established through the injection of NG-nitro-l-arginine methyl ester (L-NAME). Furthermore, related experiments were performed to detect cellar phenotype-associated signalling pathway, including epithelial-mesenchymal transition (EMT) and Wnt/ß-catenin. Cell assays indicated that RE could increase trophoblasts migration and invasion. In addition, hypertension and proteinuria were markedly ameliorated by RE compared with the controls in PE mice model. Moreover, treatment by RE in trophoblasts or in PE model, we found that RE activated EMT progress through the regulation of E-cadherin, ß-catenin, N-cadherin, vimentin expression, and further altered the WNT-related gene expression, including WNT1, WNT3 and WNT5B. Our findings demonstrated that RE might stimulate the invasive capability of human trophoblasts by promoting EMT and mediating the Wnt/ß-catenin pathway in PE.

[Colonization of group B streptococcus in the semen of chronic prostatitis patients of childbearing age and its influence on perinatal outcomes].

OBJECTIVE: To investigate the colonization of group B streptococcus (GBS) in the semen of chronic prostatitis patients of childbearing age and its influence on perinatal outcomes. METHODS: This retrospective study included 592 cases of chronic prostatitis and another 472 non-prostatitis healthy males as controls. We collected semen samples from the subjects for bacterial and Ureaplasma urealyticum (UU) culture and quantitative fluorescence PCR analysis of chlamydia trachomatis (CT) and GBS. According to the results of culture, we divided the patients into a GBS-positive and a GBS-negative group and compared the perinatal outcomes among different groups of subjects. RESULTS: The rate of GBS colonization in the semen of the chronic prostatitis patients was 11.8% (70/592). Bacteria were detected in the semen of 54.4% of the patients (322/592), mainly including GBS (21.7% ï¼»70/322ï¼½) and E coli (19.9% ï¼»64/322ï¼½), and in 7.8% of the healthy controls (37/472), Staphylococcus aureus comprising 83.8% (31/37), with statistically significant difference in the rate of bacteria detection between the two groups (P < 0.01). The incidence rate of adverse perinatal outcomes in the cases of successful pregnancy was significantly higher in the GBS-positive (32.8% ï¼»19/58ï¼½) than in the GBS-negative (22.0% ï¼»29/132ï¼½) and the healthy control group (2.2% ï¼»6/271ï¼½) (P < 0.05). CONCLUSIONS: The rate of GBS colonization is significantly increased in the semen of chronic prostatitis patients of childbearing age, and so is the incidence of adverse perinatal outcomes in the spouses of GBS-positive males. Importance should be attached to normalized screening of GBS in chronic prostatitis patients and to standardized prevention and intervention as well.

Promotion of trophoblast invasion by lncRNA MVIH through inducing Jun-B.

Preeclampsia (PE), a pregnancy-specific disorder, is associated with impaired uterine spiral artery remodelling, which is related to the dysfunction of trophoblast cells. Lately, mounting evidence has indicated that aberrant expression of long non-coding RNAs (lncRNAs) is associated with various human diseases. The lncRNA MVIH transcript has been shown to decrease the severity of several diseases. However, the biological function of MVIH, which is down-regulated in placental tissues in PE, has not yet been clarified. Here, we report that MVIH may act as a vital factor in the pathogenesis of PE. In this study, functional analysis revealed that the silencing of MVIH expression via transfection with small interfering RNA (siRNAs) inhibited cell growth, migration, invasion, and angiogenesis in various trophoblast cell lines, and stimulation with MVIH could promote these functions. Mass spectrometry analysis revealed that MVIH could modulate Jun-B protein expression, which has been reported to potentially regulate cell growth and angiogenesis. Further cotransfection assays were performed, revealing that MVIH and Jun-B have a synergistic effect on the regulation of angiogenesis and cell proliferation. Taking these findings together, MVIH could be associated with PE and may be a candidate biomarker for its diagnosis and treatment.

The long non-coding RNA PVT1 represses ANGPTL4 transcription through binding with EZH2 in trophoblast cell.

Despite progress in diagnostics and treatment for preeclampsia, it remains the foremost cause of maternal and foetal perinatal morbidity and mortality worldwide. Over recent years, various lines of evidence have emphasized long non-coding RNAs (lncRNAs) which function as an innovative regulator of biological behaviour, as exemplified by proliferation, apoptosis and metastasis. However, the role of lncRNAs has not been well described in preeclampsia. Here, we identified a lncRNA, PVT1, whose expression was down-regulated in qRT-PCR analyses in severe preeclampsia. The effects of PVT1 on development were studied after suppression and overexpression of PVT1 in HTR-8/SVneo and JEG3 cells. PVT1 knockdown notably inhibited cell proliferation and stimulated cell cycle accumulation and apoptosis. Exogenous PVT1 significantly increased cell proliferation. Based on analysis of RNAseq data, we found that PVT1 could affect the expression of numerous genes, and then investigated the function and regulatory mechanism of PVT1 in trophoblast cells. Further mechanistic analyses implied that the action of PVT1 is moderately attributable to its repression of ANGPTL4 via association with the epigenetic repressor Ezh2. Altogether, our study suggests that PVT1 could play an essential role in preeclampsia progression and probably acts as a latent therapeutic marker; thus, it might be a useful prognostic marker when evaluating new therapies for patients with preeclampsia.

Down-regulation of LHPP in cervical cancer influences cell proliferation, metastasis and apoptosis by modulating AKT.

Cervical cancer is a leading severe malignancy throughout the world. Though various pathologies associated with cervical cancer progression have been demonstrated, further study is still necessary to reveal the tumorigenesis of cervical cancer. The protein histidine phosphatase LHPP is reported as a tumor suppressor. Histidine phosphorylation, also known as hidden phosphoproteome, is a poorly characterized post-translational modification of proteins. LHPP is evolutionarily conserved from worm to human. In the present study, we discovered that LHPP expression levels were lower in human cervical cancer tumors than that in adjacent normal tissue samples. LHPP expression levels were also reduced in several cervical cancer cell lines. Further, LHPP over-expression reduced the cell proliferation, migration and invasion, associated with the change of p53 and metastasis signaling pathways. Moreover, over-expressing LHPP markedly induced apoptosis in human cervical cancer cells via promoting the cleaved Caspse-3 and PARP. Importantly, we found that LHPP over-expression blocked AKT activation. Elevating AKT activity could abolish the role of LHPP over-expression in reducing cell proliferation and metastasis, as well as in inducing apoptotic response. Moreover, suppressing p53 expression with its inhibitor of PFTα abrogated the activity of LHPP to impede cell proliferation and metastasis, and to trigger apoptosis. AKT phosphorylation also restrained p53 expression levels in cervical cancer cells. In vivo, the anti-cervical cancer effects of LJPP were verified, which were also via the repression of cell proliferation and metastasis, and the induction of apoptosis. Therefore, LHPP could be considered as an effective candidate to develop effective therapeutic strategy against cervical cancer development.

Effect of lipoprotein-associated phospholipase A2 inhibitor on insulin resistance in streptozotocin-induced diabetic pregnant rats.

This paper aims to investigate the influence of lipoprotein-associated phospholipase A2 (Lp-PLA2) inhibitor, darapladib, on insulin resistance (IR) in streptozotocin (STZ)-induced diabetic pregnant rats. The rat models were divided into Control (normal pregnancy), STZ + saline (STZ-induced diabetic pregnant rats), STZ + Low-dose and STZ + High-dose darapladib (STZ-induced diabetic pregnant rats treated with low-/high-dose darapladib) groups. Pathological changes were observed by Hematoxylin-eosin (HE) and Immunohistochemistry staining. Lp-PLA2 levels were determined by enzyme-linked immunosorbent assay (ELISA). An automatic biochemical analyzer was used to measure the serum levels of biochemical indicators, and homeostatic model assessment for insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) were calculated. Western blot was applied to determine levels of inflammatory cytokines. Compared with Control group, rats in the STZ + saline group were significantly decreased in body weight, the number of embryo implantation, the number of insulin positive cells and pancreatic islet size as well as the islet endocrine cells, and high-density lipoprotein (HDL-C) level, but substantially increased in Lp-PLA2, low-density lipoprotein (LDL-C), fatty acids (FFA), serum total cholesterol (TC), triglyceride (TG) levels. Moreover, the increased fasting plasma glucose (FPG) and HOMA-IR and inflammatory cytokines but decreased fasting insulin (FINS) and ISI were also found in diabetic pregnant rats. On the contrary, rats in the darapladib-treated groups were just opposite to the STZ + saline group, and STZ + High-dose group improved better than STZ + Low-dose group. Thus, darapladib can improve lipid metabolism, and enhance insulin sensitivity of diabetic pregnant rats by regulating inflammatory cytokines.

Prenatal diagnosis of single gene disorders using amniotic fluid as the starting material for PCR.

A rapid and inexpensive method for fetal genetic diagnosis using amniotic fluid (AF) as the starting material was demonstrated in this study. Raw AF was added directly to polymerase chain reaction (PCR) mixtures with HpH buffer (a high pH buffer), without any pre-treatment. Amplified products were detected by gel electrophoresis and then subjected to Sanger sequencing. The AF from four fetuses, each expressing a single gene disorder (achondroplasia, hypochondroplasia, thanatophoric dysplasia, or X-linked hypohidrotic ectodermal dysplasia), were analyzed. DNA fragments of different lengths were efficiently amplified from 8 µl of AF, allowing each of these single gene disorders to be successfully diagnosed. Although the amplification efficiency of the AF-PCR method is comparable to that of the Chelex method, the amount of the AF sample required was considerably lower than that required for the Chelex method (10 ml). This proposed method of diagnosis is more efficient, simpler, and less expensive, and reduces the chance of cross-contamination relative to the Chelex method, which requires purified DNA or other pre-treatment processes. Our method offers a promising tool that can be used for the diagnosis of various gene disorders in fetuses.

Non-biased and efficient global amplification of a single-cell cDNA library.

Analysis of single-cell gene expression promises a more precise understanding of molecular mechanisms of a living system. Most techniques only allow studies of the expressions for limited numbers of gene species. When amplification of cDNA was carried out for analysing more genes, amplification biases were frequently reported. A non-biased and efficient global-amplification method, which uses a single-cell cDNA library immobilized on beads, was developed for analysing entire gene expressions for single cells. Every step in this analysis from reverse transcription to cDNA amplification was optimized. By removing degrading excess primers, the bias due to the digestion of cDNA was prevented. Since the residual reagents, which affect the efficiency of each subsequent reaction, could be removed by washing beads, the conditions for uniform and maximized amplification of cDNAs were achieved. The differences in the amplification rates for randomly selected eight genes were within 1.5-folds, which could be negligible for most of the applications of single-cell analysis. The global amplification gives a large amount of amplified cDNA (>100 µg) from a single cell (2-pg mRNA), and that amount is enough for downstream analysis. The proposed global-amplification method was used to analyse transcript ratios of multiple cDNA targets (from several copies to several thousand copies) quantitatively.