Common occurrence of hotspots of single strand DNA breaks at transcriptional start sites.

BACKGROUND: We recently developed two high-resolution methods for genome-wide mapping of two prominent types of DNA damage, single-strand DNA breaks (SSBs) and abasic (AP) sites and found highly complex and non-random patterns of these lesions in mammalian genomes. One salient feature of SSB and AP sites was the existence of single-nucleotide hotspots for both lesions. RESULTS: In this work, we show that SSB hotspots are enriched in the immediate vicinity of transcriptional start sites (TSSs) in multiple normal mammalian tissues, however the magnitude of enrichment varies significantly with tissue type and appears to be limited to a subset of genes. SSB hotspots around TSSs are enriched on the template strand and associate with higher expression of the corresponding genes. Interestingly, SSB hotspots appear to be at least in part generated by the base-excision repair (BER) pathway from the AP sites. CONCLUSIONS: Our results highlight complex relationship between DNA damage and regulation of gene expression and suggest an exciting possibility that SSBs at TSSs might function as sensors of DNA damage to activate genes important for DNA damage response.

Genome-wide profiles of DNA damage represent highly accurate predictors of mammalian age.

The identification of novel age-related biomarkers represents an area of intense research interest. Despite multiple studies associating DNA damage with aging, there is a glaring paucity of DNA damage-based biomarkers of age, mainly due to the lack of precise methods for genome-wide surveys of different types of DNA damage. Recently, we developed two techniques for genome-wide mapping of the most prevalent types of DNA damage, single-strand breaks and abasic sites, with nucleotide-level resolution. Herein, we explored the potential of genomic patterns of DNA damage identified by these methods as a source of novel age-related biomarkers using mice as a model system. Strikingly, we found that models based on genomic patterns of either DNA lesion could accurately predict age with higher precision than the commonly used transcriptome analysis. Interestingly, the informative patterns were limited to relatively few genes and the DNA damage levels were positively or negatively correlated with age. These findings show that previously unexplored high-resolution genomic patterns of DNA damage contain useful information that can contribute significantly to both practical applications and basic science.

Triglyceride-glucose index predicts the risk of gestational diabetes mellitus: a systematic review and meta-analysis.

AIMS: We aimed to investigate the potential predictive efficacy of triglyceride-glucose (T/Gly) index for gestational diabetes mellitus (GDM) in a systematic review and meta-analysis. MATERIALS AND METHODS: Cohort studies demonstrating the association between T/Gly index measured at the first trimester or before pregnancy and the subsequent incidence of GDM were identified by search of PubMed, Embase, China National Knowledge Infrastructure, and WanFang databases. A random-effect model incorporating the heterogeneity was applied to pool the results. Five cohort studies including 382,213 women were included in this meta-analysis. RESULTS: Compared to those with the lowest category of T/Gly index, women with the highest category of T/Gly index were independently associated with higher risk of subsequent GDM (odds ratio [OR]: 2.52, 95% confidence interval [CI]: 1.33 ∼ 4.67, I2=65%, p=.004) after adjustment of potential confounding factors including age, body mass index, and family history of diabetes. The association was stronger in prospective cohort studies than retrospective cohort studies (p for subgroup difference=.007), and a significant association was detected in Asian women (OR: 3.30, 95% CI: 1.50-7.28, p =.003), but not in non-Asian women (OR: 0.96, 95% CI: 0.35-2.63, p=.94). CONCLUSIONS: Higher T/Gly index may independently predict the risk of GDM in Asian women.

Dysregulated miRNAs contribute to altered placental glucose metabolism in patients with gestational diabetes via targeting GLUT1 and HK2.

INTRODUCTION: Dysregulated genes in glucose transport and metabolize pathways have been found in patients with Gestational diabetes (GDM), but the underlying mechanisms were still unclear. MATERIALS AND METHODS: Placental villous samples were collected from 31 patients with GDM and 20 healthy controls. The expression of GLUT1, GLUT4, GLUT9 and HK2 was examined by immunoblotting and qRT-PCR. The miRNAs have the potential targeting GLUT1 and HK2 were predicted using online bioinformatics tool: TargetScan. The interaction between miRNAs and target genes were confirmed by dual luciferase assay and immunoblotting. The function of miR-9 and miR-22 on glucose metabolism was examined by glucose uptake assay and lactate secretion assay. RESULTS: GLUT1 and HK2 proteins level was found upregulated in patients with GDM, but the mRNA level was not significantly changed. Predicted by using bioinformatics tools and confirmed by dual luciferase assay and immunoblotting, GLUT1 was identified as a target of miR-9 and miR-22, whereas HK2 was identified as a target of miR-9. MiR-9 and miR-22 level was found reduced in the placenta villous and negatively correlated with the expression of GLUT1 and HK2. Functional studies indicated that miR-9 and miR-22 inhibitors upregulated the expression of GLUT1 and HK2, and then increased the glucose uptake, lactate secretion, cell viability and repressed apoptosis in primary syncytiotrophoblasts (STBs) and HTR8/SVneo cells. DISCUSSION: The upregulation of GLUT1 and HK2 in the placenta, which is induced by miR-9 and miR-22 reduction, contributes to the disordered glucose metabolism in patients with GDM.

The developmental pattern of the RAS/RAF/Erk1/2 pathway in the BTBR autism mouse model.

BTBR mice exhibit several autistic-like behaviors and are currently used as a model for understanding mechanisms that may be responsible for the pathogenesis of autism. Ras/Raf/ERK1/2 signaling has been suggested to play an important role in neural development, learning, memory, and cognition. Two studies reported that a deletion of a locus on chromosome 16 containing the mitogen-activated protein kinase 3 (MAPK3) gene, which encodes ERK1, is associated with autism. In the present study, Ras/Raf/ERK1/2 signaling was found to be up-regulated in BTBR mice relative to matched control B6 mice, to further suggest involvement in the pathogenesis of autism. To further characterize the developmental pattern of Ras/Raf/ERK1/2 signaling, varying stages during development were sampled to reveal an up-regulation in newborn and 2-week old BTBR mice relative to age-matched B6 mice. By the age of 3-week, Ras/Raf/ERK1/2 signaling in the brain of BTBR mice was unaltered relative to B6 mice, with this trend maintained in 6-week samples. These results suggest that the alteration of Ras/Raf/ERK signaling in the early developmental stages in mice could contribute to the noted autistic phenotype. Furthermore, these findings support the value of BTBR mice to serve as a human analog for autistic etiological research and aid in a better understanding of the developmental mechanisms of autism.

[Prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong province].

OBJECTIVE: To investigate the prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong province. METHODS: Data from 169 218 pregnant women in different regions of Guangdong province from January 2005 to June 2010 were analyzed retrospectively. The prevalence and epidemiological characteristics of thromboembolic disease during pregnancy or puerperium were investigated. RESULTS: Of the studied population, (1) 201 cases (1.3‰) suffered from thromboembolic disease during pregnancy or puerperium including 128 cases of deep vein thrombosis (DVT), 68 cases of cerebral venous thrombosis (CVT) and 5 pulmonary embolism, the prevalence rates were 0.8‰, 0.4‰, and 0.02‰ respectively. (2) Risk factors in different regions showed that, in the Pearl River Delta area, the major risk factors for DVT would include previous or family history of thrombosis, pregnancy complications, with medically involved diseases, prolonged bed rest and pregnancy weight gain>15 kg etc. While in eastern, western, northern parts of Guangdong, the major risk factors for DVT would include pregnancy weight gain>15 kg, prolonged bed rest, preeclampsia, cesarean section and complications during pregnancy. In Pearl River Delta region, the major risk factors for CVT would include eclampsia, preeclampsia, pregnancy complications, prolonged bed rest>3 days, past history or family history of thrombosis. While eclampsia, preeclampsia, advanced age or younger age, pregnancy weight gain>15 kg, complications during pregnancy were the major risk factors for CVT in the eastern, western or northern parts of Guangdong. CONCLUSION: Prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong were different. It was crucial to take effective measures in pregnant women with different epidemiological characteristics and risk factors to prevent and reduce the incidence of peripartum thromboembolic disease.

microRNA regulation of the expression of the estrogen receptor in endometrial cancer.

microRNAs (miRNAs) are non-coding RNA transcripts thought to be instrumental in controlling eukaryotic cell function. This study aimed to identify miRNAs associated with the differential expression of estrogen receptor α (ERα) and progesterone receptor (PR) in uterine endometrial cancer. First, the pathology of Ishikawa and KLE cell lines was identified by transplant and biopsy. Expression levels of ERα and PR and their response to estrogen and progesterone were examined, and total RNA was isolated to identify differentially expressed miRNAs by microarray analysis. miRNAs targeting ESR1 and PGR were predicted by miRANDA and TargetScan software, and their expression levels in cell lines and in patient samples were validated by real-time RT-PCR. Ishikawa and KLE represent estrogen-dependent and estrogen-independent endometrial cancers, respectively. As identified by miRNA microarray, 126 miRNAs were differentially expressed between the two cell lines; hsa-miR-100 and hsa-miR-99a were predicted to target ESR1, and hsa-miR-378 and hsa-miR-768-3p to target PGR. The differential expression of these miRNAs between Ishikawa and KLE was consistent in vivo and in vitro. Hsa-miR-100 was significantly down-regulated in the estrogen-dependent endometrial cancer samples as compared to the estrogen-independent samples and thus has the potential to target ESR1.

Pharmacokinetic comparison between systemic and local chemotherapy by carboplatin in dogs.

The aim of the current study was to compare pharmacokinetics of local chemotherapy by pelvic intra-arterial administration with intravenous injection in dogs. A total of 18 female dogs (weight: 10-15 kg) were randomly divided into 3 groups: the peripheral vein administration (group A, n = 6), the abdominal aorta administration (group B, n = 6), and the internal iliac artery administration (group C, n = 6). Carboplatin at a dose of 1.2 mg/kg was administered by infusing into the arteries or the vein. For analysis, plasma and uterine tissue samples were collected at different times following infusion. The peak local concentration of platinum in the uterus of dogs in group C was significantly higher than those of groups A and B (P < .05). The area under the tissue concentration-time curve (AUC) of uterine tissues was significantly higher in dogs of group C compared to those of the other 2 groups (P < .05). There was no significant difference in the AUC of the uterine tissues of dogs between groups A and B (P > .05). The peak concentration of platinum in plasma was significantly higher in group A compared to those of the other 2 administration routes (P < .05). We observed the pharmacokinetic advantages of local chemotherapy by internal iliac artery perfusion with the chemotherapeutic agent, carboplatin, to the uterus, thereby leading to a high-drug concentration that may be more effective in treating cervical cancer.

[Expression of ZNF217 in human ovarian cystadenocarcinoma and its clinical significance].

OBJECTIVE: To explore the correlation of ZNF217 expression to the carcinogenesis and progression of human ovarian cancer. METHODS: Immunohistochemistry and real-time RT-PCR were used to detect ZNF217 expression in human ovarian cystadenocarcinoma, ovarian cystadenoma and normal ovary tissues. RESULTS: The expression levels of ZNF217 protein and mRNA in ovarian cystadenocarcinoma was significantly higher than those in matched ovarian cystadenoma and normal tissues (P<0.05). No significant difference was found in the expression between ovarian cystadenoma and normal ovarian tissues (P>0.05). The mRNA expression in the specimens was consistent with the protein expression of ZNF217 (P<0.05). CONCLUSION: ZNF217 gene expression is closely correlated to the occurrence and clinical stages of ovarian carcinomas, suggesting that ZNF217 can be an important candidate gene responsible for the occurrence and progression of ovarian carcinomas.

[Ultrasonic study of deep-vein diameter and blood flow spectrum changes in full-term pregnant women].

OBJECTIVE: To explore the changes in lower limb deep vein diameters, blood flow velocity and blood biochemistry in full-term pregnant women for early diagnosis and treatment of prothrombotic state. METHODS: One hundred and twenty-eight full-term pregnant women at high risk of thrombosis (Group A), 61 healthy full-term pregnant women (Group B), and 42 healthy non-pregnant women (Group C) underwent high-resolution color Doppler ultrasound (CDU) for examining the deep veins of the lower limbs. The hematological indexes such as D-D, PLT, HGB, HCT, TT, APTT, PT, and FbgC were also observed in these 3 groups. RESULTS: Compared to Group B, the women in group A showed significantly increased diameters of the common femoral veins (CFV) and left superficial femoral vein (SFV), HCT and DD, but with significantly decreased peak blood flow in the bilateral popliteal veins (POPV) (P<0.01) and increased left POPV diameter (P=0.034). Compared to those in group C, the diameters of the bilateral CFVs, SFVs, POPV, and posterior tibial veins (PTVs) were significantly increased, but the peak blood flow in the bilateral CFVs and POPVs were significantly reduced in groups A and B; the PLT, HGB, HCT, DD, TT, APTT, PT, and FbgC also showed significant changes in groups A and B (P<0.01). CONCLUSION: The full-term pregnant women are at higher risk of prothrombotic state than non-pregnant women, and the full-term pregnant women with the high risk factors for thrombosis are more likely to have prothrombotic state than healthy full-term pregnant women. CDU examination of the lower limb deep veins can be of value in the diagnosis of prothrombotic state.

Pharmacokinetic comparison between pelvic transarterial chemoembolization and transcatheter arterial chemotherapy in an animal model.

Transcatheter arterial chemotherapy (TAC) and transarterial chemoembolization (TACE) have greatly benefited patients suffering from gynecologic malignancies. In cervical cancer treatment, TACE appears to be more effective than TAC. We compared the pharmacokinetics of TACE with those of TAC in an animal study. Twelve female dogs were randomly divided into 2 groups (n=6), and carboplatin was applied at a concentration of 1.2 mg/kg. Carboplatin and lipiodol-carboplatin were administered by perfusion into the bilateral internal iliac arteries of the animals in the two groups, respectively. Plasma and tissues from the ovaries and uterus were collected for analysis at various time points after TAC or TACE. Pharmacokinetic analysis indicated that the uterine tissue area under the concentration (AUC) of carboplatin was approximately 2 times higher in the TACE group than in the TAC group (P<0.01). By contrast, the ovarian tissue AUC of carboplatin was much lower in the TACE group than in the TAC group (P<0.01), and was much lower than the uterine tissue AUC (P<0.01) in both groups. The plasma AUC of carboplatin was more than 2 times higher in the TACE group than in the TAC group (P<0.01). In conclusion, a pharmacological advantage of TACE treatment with lipiodol-carboplatin for gynecologic malignancies was observed.

[Hysterectomy after cardiopulmonary resuscitation in patients with obstetric hemorrhagic shock].

OBJECTIVE: To explore the indication of hysterectomy after successful resuscitation of cardiac arrest due to obstetric hemorrhagic shock. METHODS: A retrospective analysis was conducted in 13 patients with cardiac arrest due to obstetric hemorrhagic shock in 7 hospitals of Guangzhou, including 12 patients undergoing hysterectomy and 1 undergoing uterine artery embolization. RESULTS: s After successful cardiopulmonary resuscitation, only 4 of the 13 patients undergoing hysterectomy or uterine artery embolization for continuing uterus hemorrhage survived. CONCLUSION: Detailed plans and emergency measures should be formulated in the management of high-risk pregnancies. Early diagnosis and active treatment of obstetric hemorrhagic shock with hysterectomy or uterine artery embolization are critical in preventing cardiac arrest and improving the survival of the patients.

[Clinical study of Osaka formula and improved multiparameter ultrasonic measurement for fetal weight estimation].

OBJECTIVE: To analyze the factors affecting the accuracy of Osaka formula multiparameter ultrasound-based fetal mass estimation, thereby establishing new formulas to improve the accuracy of the estimation. METHODS: A retrospective review was conducted among 519 healthy women with singleton pregnancy. Three days before the delivery (between 37 and 42 weeks' gestation), ultrasonic measurement of the fetal weight and other indices of the fetus was routinely performed. Correlation and multiple linear stepwise regression analysis were used to correct the 3 equations, which, along with Osaka University formula, were used to predict another 219 fetuses' birth weight. The coincidence rate of the predicted value and with the actual birth weight, and the absolute error and relative error were compared between the equations. RESULTS: The fetal abdominal area (AA) and abdominal circumference (AC) showed the most conspicuous influence on the estimated fetal birth weight, and fetal humerus length (HL) was more sensitive than femur length (FL) for the estimation. Three new regression equations were established, among which the equation 2 (fetal birth weight=1082.859+4.116xAAxHL) showed the best accuracy in clinical prediction. CONCLUSION: AA,AC and HL are more sensitive indices for estimation of the fetal birth weight, and the equation 2 established in this study still awaits further verification for its clinical value.

[Changes in serum endothelin and in pregnant rabbits with pulmonary embolism].

OBJECTIVE: To study change of serum endothelin (ET) and in serum endothelin and in pregnant rabbit with acute pulmonary embolism. METHODS: In 17 pregnant rabbits, autologous blood clots were administered into the right atrium via the femoral vein to establish the animal models of acute pulmonary embolism, and another 10 pregnant rabbits received injection with saline to serve as the control group. Serum levels of ET and CGRP were measured before, 0, 2, 4, 6 and 8 h after the injection. RESULTS: Acute pulmonary embolism was induced in all the rabbits receiving the injection of autologous blood clots with a success rate of 100%. In these rabbit models of pulmonary embolism, serum ET increased 0 h after the embolism and kept rising till 6 h, whereas CGRP level underwent immediate decrease after the embolism followed then by increase during the period from 4 to 8 h, showing significant difference from the levels of the control group. CONCLUSION: Significant difference in serum ET and CGRP levels occur in the event of acute pulmonary embolism in pregnant rabbits.