Age and menopausal status are important factors influencing the serum human epididymis secretory protein 4 level: a prospective cross-sectional study in healthy Chinese people / 中华医学杂志(英文版)

BACKGROUND@#Human epididymis secretory protein 4 (HE4) is a new ovarian cancer biomarker. The factors influencing HE4 levels are not clear, and the reference data in China are limited. Here, we aim to evaluate the effects of menopause and age on HE4 levels and to provide a possible reference value for HE4 in healthy Chinese people.@*METHODS@#A total of 2493 healthy females aged 40 years or older were recruited from March 2013 to March 2017 with the cooperation of four medical institutions across Beijing, China. The serum levels of HE4 and cancer antigen 125 (CA125) were measured by enzyme-linked immunosorbent assay. The Wilcoxon rank-sum test of variance and a stratified analysis were used to analyze the relationships among age, menopausal status, and levels of HE4 or CA125. Confidence intervals (5%-95%) were determined for reference ranges in different populations.@*RESULTS@#There was a statistically significant difference in median HE4 levels between the post-menopausal (n = 2168) and pre-menopausal groups (n = 325) (36.46 vs. 24.04 pmol/L, Z = -14.41, P < 0.001). HE4 increased significantly with age in the post-menopausal groups (H = 408.18, P < 0.001) but not in the pre-menopausal subjects (Z = -0.43, P = 0.67). The upper 95th percentile of HE4 levels were 44.63 pmol/L for pre-menopausal women, 78.17 pmol/L for post-menopausal women, and 73.3 pmol/L for all women. In the post-menopausal population, the HE4 reference ranges were 13.15 to 47.31, 14.31 to 58.04, 17.06 to 73.51, 24.50 to 115.25, and 35.71 to 212.37 pmol/L for different age groups from forty divided by decade. The CA125 level was affected mainly by menopausal status and not age.@*CONCLUSIONS@#Menopausal status and age were both important factors influencing the level of HE4, and age affected HE4 levels mainly in post-menopausal women. The HE4 level was higher in the post-menopausal population than in the pre-menopausal population and increased with age.

Evaluation of Circulating Endometrial Cells as a Biomarker for Endometriosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Circulating endometrial cells (CECs) have been reported to be present in the peripheral blood of women with endometriosis (EM), providing clear and specific evidence of the presence of ectopic lesions. In this study, we established a method with a high detection rate of CECs, assessed the diagnostic value of CECs for EM and compared with serum CA125, and proposed a hypothesis for the pathogenesis of EM from the new perspective of CECs.</p><p><b>METHODS</b>The participants were enrolled prospectively from October 2015 to July 2016. The peripheral blood samples were collected from 59 participants, and the blood cells were isolated for immunofluorescence staining via microfluidic chips. The cells that were positive for vimentin/cytokeratin and estrogen/progesterone receptor and negative for CD45 were identified as CECs. The serum CA125 level was tested with electrochemiluminescence immunoassay.</p><p><b>RESULTS</b>The detection rate of CECs reached 89.5% (17/19) in the EM group, which was significantly higher than that of the control group (15.0% [6/40], P < 0.001) and was independent of menstrual cycle phases. Furthermore, a positive CEC assay detected 4/5 cases of Stage I-II EM. In contrast, a positive CA125 test had limited value in detecting EM (13/19, 68.4%) and detected only one case of Stage I-II EM.</p><p><b>CONCLUSION</b>CECs are promising biomarkers for EM with great potential for a noninvasive diagnostic assay.</p>

Low programmed cell death 5 expression is a prognostic factor in ovarian cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Ovarian cancer is a leading gynecological malignancy. We investigated the prognostic value of programmed cell death 5 (PDCD5) in patients with ovarian cancer.</p><p><b>METHODS</b>Expression levels of PDCD5 mRNA and protein were examined in six ovarian cancer cell lines (SKOV3, CAOV3, ES2, OV1, 3AO, and HOC1A) and one normal ovarian epithelial cell line (T29) using reverse transcription polymerase chain reaction, Western blotting, and flow cytometry. After inducing PDCD5 induction in SKOV3 cells or treating this cell line with taxol or doxorubicin (either alone or combined), apoptosis was measured by Annexin V-FITC/propidium iodide staining. Correlations between PDCD5 protein expression and pathological features, histological grade, FIGO stage, effective cytoreductive surgery, and serum cancer antigen-125 values were evaluated in patients with ovarian cancer.</p><p><b>RESULTS</b>PDCD5 mRNA and protein expression were downregulated in ovarian cancer cells. Recombinant human PDCD5 increased doxorubicin-induced apoptosis in SKOV3 cells (15.96 ± 2.07%, vs. 3.17 ± 1.45% in controls). In patients with ovarian cancer, PDCD5 expression was inversely correlated with FIGO stage, pathological grade, and patient survival (P < 0.05, R = 0.7139 for survival).</p><p><b>CONCLUSIONS</b>PDCD5 expression is negatively correlated with disease progression and stage in ovarian cancer. Therefore, measuring PDCD5 expression may be a good method of determining the prognosis of ovarian cancer patients.</p>

Effect of high-fat diet on liver and placenta fatty infiltration in early onset preeclampsia-like mouse model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Preeclampsia, especially early onset of preeclampsia (PE), is a common and serious disorder with high maternal and perinatal morbidity and mortality. Dietary factor is one of the most important factors which may affect the occurrence and development of the disease. The aim of this study is to investigate the effects of dietary factors on pathological changes of liver and placenta in preeclampsia-like mouse model by establishing the model at multiple stages of gestation.</p><p><b>METHODS</b>Wild-type (WT) mice were injected subcutaneously with nitric oxide synthase (NOS) inhibitor L-arginine methyl ester (L-NAME, 50 mg×kg(-1)×d(-1)) to establish PE-like model (L-NAME group) at early-, mid-, and late-pregnant periods respectively; simultaneously, the control mice were injected with normal saline (NS group). All the groups were divided into subgroups, standard chow group (SC), and high-fat diet group (HF). ApoE(-/-) pregnant mice served as a control group. Systolic blood pressure (SBP), urine protein, and histopathologic changes of placenta and liver in all groups were observed and statistically analyzed.</p><p><b>RESULTS</b>In WT and apoE(-/-) L-NAME subgroups, blood pressure and urine protein were significantly higher than those in all the gestational age matched NS groups (P < 0.05). Compared to other groups, remarkable liver fatty infiltration and lipid storage in placenta were found in early- and mid-L-NAME subgroups in apoE(-/-) mice (P < 0.05), especially in the early- and mid-HF+L-NAME subgroups in apoE(-/-) mice (P < 0.05). More lipid storage droplets both in liver and placenta were found in ApoE(-/-) mice than that of WT groups (P < 0.05). Morphology histopathologic examination of placentas showed varying degrees of fibrinoid necrosis and villous interstitial edema in early- and mid-L-NAME both in HF and SC of apoE(-/-) and WT subgroups compared to NS controls (P < 0.05). But there was no significant difference between HF and SC subgroups (P > 0.05), and no difference between apoE(-/-) and WT groups (P > 0.05).</p><p><b>CONCLUSIONS</b>Preeclampsia-like conditions could be induced by L-NAME in mice at different gestational stages. Both WT and apoE(-/-) genotype mice with preeclampsia-like symptoms in early and mid stages of pregnancy presented lipid deposition in the placenta and hepatic fatty infiltration. To alter the environmental condition by feeding high-fat diet was harmful to the mother and the fetus. High-fat diet aggravated the impact of liver fatty infiltration at early and mid gestational stages especially in the apoE(-/-) mouse model. These results further revealed the association between early-onset preeclampsia and the dysoxidation of fatty acids.</p>

Inhibition of nitric oxide synthase lowers fatty acid oxidation in preeclampsia-like mice at early gestational stage / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Preeclampsia is one of hypertensive disorders in pregnancy. It is associated with abnormal lipid metabolism, including fatty acid oxidation metabolism. Long chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) plays an indispensable role in the oxidation of fatty acids. It has been reported that nitric oxide (NO) is one of the regulatory factors of the fatty acid oxidation pathway. The aim of this research was to investigate whether the nitric oxide synthase (NOS) inhibitor L-NAME may cause down-regulation of LCHAD in the pathogenesis of preeclampsia.</p><p><b>METHODS</b>Pregnant wild-type (WT) mice were treated with L-NAME or normal saline (NS) during gestation days 7 - 18 (early group), days 11 - 18 (mid group) and days 16 - 18 (late group), and apoE-/- mice served as a control. Systolic blood pressure (SBP), urine protein, feto-placental outcome, plasma lipid levels and NO concentrations were measured, and the expression of mRNA and protein for LCHAD in placental tissue were determined by real-time polymerase chain reaction (RT-PCR) and Western blotting, respectively.</p><p><b>RESULTS</b>In WT and apoE-/- mice, SBP and urinary protein increased following L-NAME injection. Fetal and placental weights and NO concentrations were reduced and total cholesterol, triglycerides and free fatty acid levels were increased in early and mid L-NAME groups in WT and apoE-/- mice, compared with the NS group. There was no significant difference between the late L-NAME group and NS group. RT-PCR and Western blotting analysis showed that the mRNA and protein levels of LCHAD expression were significantly down-regulated in the early and mid L-NAME groups but not in the late L-NAME group in the WT and apoE-/- mice compared with the corresponding NS groups.</p><p><b>CONCLUSIONS</b>Inhibition of NO in early and mid gestation in mice may cause hyperlipidemia and suppression of fatty acid oxidation, whereas preeclampsia-like conditions in late gestation may be a maternal vascular response to inhibition of NO.</p>

Effects of preeclampsia-like symptoms at early gestational stage on feto-placental outcomes in a mouse model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Early and late-onset preeclampsia is thought to be different disease entities. This study aimed to determine the effects of early-onset preeclampsia-like symptoms on feto-placental outcomes and the adverse impacts of various factors on placental and fetal growth and development at different gestational stages in a mouse model.</p><p><b>METHODS</b>Pregnant C57BL/6J mice were divided into control and preeclampsia (PE) groups, and injected subcutaneously with the nitric oxide synthase inhibitor L-arginine methyl ester (L-NAME) 50 mgxkg(-1)d(-1). The PE group was divided into early-, mid- and late-PE groups with L-NAME injections starting on days 7, 11 and 16 of pregnancy, respectively. Corresponding control groups were injected with saline at the same time points. Blood pressure was measured until days 14 and 18, when the fetuses and placentas were removed under anesthesia. Blood pressure, urinary protein, and fetal and placental conditions were analyzed.</p><p><b>RESULTS</b>Blood pressure and urinary protein increased following L-NAME injection. The fetal survival rate and fetal weight were reduced and the fetal absorption rate was increased in the early-PE group on days 14 and 18 of pregnancy, compared with the control group. There were no significant differences in these parameters between the late-PE group and the respective control group. Placental weights in the early- and mid-PE groups were significantly reduced at days 14 and 18 of pregnancy compared with the control groups, but there was no significant difference in placental weight between the late-PE group and the respective control group. Morphologic examination of placentas from the early- and mid-PE groups showed varying degrees of fibrinoid necrosis and villous interstitial edema, but no significant pathologic changes were found in the placentas from the late-PE or control groups.</p><p><b>CONCLUSION</b>Preeclampsia-like symptoms occurring during the early stage of pregnancy are more likely to affect placental and fetal development, whereas late onset preeclampsia-like symptoms have a direct impact on the mothers.</p>

Scanning electron microscopic observation: three-dimensional architecture of the collagen in hepatic fibrosis rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In the process of hepatic fibrosis, the accumulation of collagen fibers is strongly related to the hepatic function. The aim of this study was to investigate the three-dimensional architecture of the collagen network in the liver of rats with hepatic fibrosis.</p><p><b>METHODS</b>Healthy adult male Wistar rats (n = 32) were randomly divided into a control group (n = 16) and a hepatic fibrosis group (n = 16). In the control group, the rats were treated with peanut oil while the rats in hepatic fibrosis group were treated for 10 weeks with 60% CCl(4) diluted in peanut oil. The quantity of collagen fibers was detected by Western blotting; distribution of the collagen was detected by sirius red staining and polarized microscope; the three-dimensional architecture of collagen in the liver was observed under the scanning electron microscope after fixed tissues were treated with cell-maceration using NaOH. Statistical analysis was performed using the u test.</p><p><b>RESULTS</b>The quantity of collagen fibers increased significantly in the hepatic fibrosis group. With the aggravation of hepatic fibrosis, collagen fibers gradually accumulated. They interlaced the reticulation compartment and formed a round or ellipse liver tissue conglomeration like a grape framework that was disparate and wrapped up the normal liver lobule. The deposition of collagen fibers was obvious in adjacent hepatic parenchyma, especially around the portal tracts.</p><p><b>CONCLUSION</b>Our experiment showed the collagen proliferation and displays clearly the three-dimensional architecture of collagen fibers in rat liver with hepatic fibrosis by scanning electron microscope. It can provide a morphological foundation for the mechanisms of changed haemodynamics and portal hypertension in hepatic fibrosis.</p>