Inhibition of Notch 1 signaling in the subacute stage after stroke promotes striatal astrocyte-derived neurogenesis.

Inhibition of Notch1 signaling has been shown to promote astrocyte-derived neurogenesis after stroke. To investigate the regulatory role of Notch1 signaling in this process, in this study, we used a rat model of stroke based on middle cerebral artery occlusion and assessed the behavior of reactive astrocytes post-stroke. We used the γ-secretase inhibitor N-[N-(3,5-diuorophenacetyl)-1-alanyl]-S-phenylglycine t-butylester (DAPT) to block Notch1 signaling at 1, 4, and 7 days after injury. Our results showed that only administration of DAPT at 4 days after stroke promoted astrocyte-derived neurogenesis, as manifested by recovery of white matter fiber bundle integrity on magnetic resonance imaging, which is consistent with recovery of neurologic function. These findings suggest that inhibition of Notch1 signaling at the subacute stage post-stroke mediates neural repair by promoting astrocyte-derived neurogenesis.

Differentiation of Treatment-Related Effects from Glioma Recurrence Using Machine Learning Classifiers Based Upon Pre-and Post-Contrast T1WI and T2 FLAIR Subtraction Features: A Two-Center Study.

PURPOSE: We propose three support vector machine (SVM) classifiers, using pre-and post-contrast T2 fluid-attenuated inversion recovery (FLAIR) subtraction and/or pre-and post-contrast T1WI subtraction, to differentiate treatment-related effects (TRE) from glioma recurrence. MATERIALS AND METHODS: Fifty-six postoperative high-grade glioma patients with suspicious progression after radiotherapy and chemotherapy from two centers were studied. Pre-and post-contrast T1WI and T2 FLAIR were collected. Each pre-contrast image was voxel-wise subtracted from the co-registered post-contrast image. Dataset was randomly split into training, and testing on a 7:3 ratio, accordingly subjected to a five fold cross validation. Best feature subsets were selected by Pearson correlation coefficient and recursive feature elimination, whereupon a radiomics classifier was built with SVM. The discriminating performance was assessed with the area under receiver-operating characteristic curve (AUC), accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). RESULTS: In all, 186 features were extracted on each subtraction map. Top nine T1WI subtraction features, top thirteen T2 FLAIR subtraction features and top thirteen combination features were selected to build optimal SVM classifiers accordingly. The accuracies/AUCs/sensitivity/specificity/PPV/NPV of SVM based on sole T1WI subtraction were 80.00%/80.00% (CI: 0.5370-1.0000)/100%/70.00%/62.50%/100%. Those results of SVM based on sole T2 FLAIR subtraction were 86.67%/84.00% (CI: 0.5962-1.0000)/100%/80%/71.43%/100%. Those results of SVM based on both T1WI subtraction and T2 FLAIR subtraction were 93.33%/94.00% (CI: 0.7778-1.0000)/100%/90%/83.33%/100%, respectively. CONCLUSION: Pre- and post-contrast T2 FLAIR subtraction provided added value for diagnosis between recurrence and TRE. SVM based on a combination of T1WI and T2 FLAIR subtraction maps was superior to the sole use of T1WI or T2 FLAIR for differentiating TRE from recurrence. The SVM classifier based on combination of pre-and post-contrast subtraction T2 FLAIR and T1WI imaging allowed for the accurate differential diagnosis of TRE from recurrence, which is of paramount importance for treatment management of postoperative glioma patients after radiation therapy.

A perfusion territory shift attributable solely to the secondary collaterals in moyamoya patients: a potential risk factor for preoperative hemorrhagic stroke revealed by t-ASL and 3D-TOF-MRA.

OBJECTIVE: The authors conducted a study to noninvasively and nonradioactively reveal moyamoya disease (MMD) intracerebral perfusion and perfusion territory supplied by the unilateral internal carotid artery (ICA) and external carotid artery (ECA) and bilateral vertebral arteries (VAs) before surgery and to further identify risk factors for preoperative hemorrhage in adult MMD. METHODS: Forty-three consecutive adult patients with bilateral MMD underwent unenhanced T1-weighted MRI, territorial arterial spin labeling (t-ASL), and unenhanced 3D time-of-flight MRA (3D-TOF-MRA). Clinical factors, including age, sex, hypertension, diabetes mellitus, hyperlipidemia, current smoking status, and history of taking aspirin, were gathered and stratified. Univariate logistic regression analyses were used to examine the relationship between various risk factors and the occurrence of preoperative hemorrhage. Stepwise multivariate logistic regression analyses were used to determine independent risk factors of preoperative hemorrhage in MMD. RESULTS: Among the 86 MMD hemispheres, t-ASL revealed 137 perfusion territory shifts in 79 hemispheres. Five distinct categories of perfusion territory shifts were observed on t-ASL maps. The subtypes of perfusion territory shift on t-ASL maps were further subdivided into 2 different categories, group A and group B, in combination with findings on 3D-TOF-MRA. A perfusion territory shift attributable solely to the secondary collaterals was a potential independent risk factor for preoperative hemorrhage (p = 0.026; 95% CI 1.201-18.615; OR 4.729). After eliminating the influence of the secondary collaterals, the primary collaterals had no significant effect on the risk of preoperative hemorrhage (p = 0.182). CONCLUSIONS: t-ASL could reveal comprehensive MMD cerebral blood perfusion and the vivid perfusion territory shifts fed by the unilateral ICA and ECA and bilateral VAs in a noninvasive, straightforward, nonradioactive, and nonenhanced manner. 3D-TOF-MRA could subdivide t-ASL perfusion territory shifts according to their shunt arteries. A perfusion territory shift attributable to the secondary collaterals is a potential independent risk factor for preoperative hemorrhage in MMD patients. A perfusion territory shift fed by the primary collaterals may not have a strong effect on preoperative hemorrhage in MMD patients. These findings make the combined modalities of t-ASL and 3D-TOF-MRA a feasible tool for MMD disease assessment, management, and surgical strategy planning.

Inhibition of Notch1 Signaling at the Subacute Stage of Stroke Promotes Endogenous Neurogenesis and Motor Recovery After Stroke.

Background and Purpose: It is still not clear whether Notch1 signaling inhibition can promote functional outcomes after stroke, given that it plays time-dependent roles in the sequential process of endogenous neurogenesis. The purpose of this study was to identify the appropriate time frame for Notch1 signaling inhibition according to the temporal evolution of Notch1 signaling activation and the responses of neural stem cells (NSCs), in order to target it for therapeutic intervention and stimulate neurorestorative strategies after stroke. Methods: Sprague-Dawley (SD) rats were subjected to 90-min of middle cerebral artery occlusion (MCAO). Rats were sacrificed before, and at day 1, day 2, day 3, day 4, and day 7 after ischemia for immunohistochemical analysis of the Notch intracellular domain (NICD), Nestin and doublecortin (Dcx). Next, MCAO rats were treated with the γ-secretase inhibitor N-[N-(3,5-di uorophenacetyl)-1-alanyl]-S-phenylglycine t-butylester (DAPT) or with saline at day 4 after ischemia, and subsequently evaluated with behavioral test analysis and magnetic resonance imaging (MRI). The rat brains were then harvested for immunohistochemical analysis of Dcx, NeuN and myelin basic protein (MBP) at 2, 3, 4, and 8 weeks. Results: Notch1 signaling was maximally activated at day 3 after ischemia in parallel with the temporal evolution of NSCs. Inhibiting Notch1 signaling at day 4 after reperfusion with DAPT further promoted recovery of MRI parameters of the corticospinal tract (CST) and the functional outcomes, concomitantly with an increase in neuroblasts, their migration to the ischemic boundary, and potential differentiation to mature neurons, as well as the amelioration of axonal bundle integrity. Conclusion: Inhibition of Notch1 signaling at the subacute stage of stroke could maximally promote endogenous neurogenesis and axonal reorganization.

Predictors of short-term outcome in patients with acute middle cerebral artery occlusion: unsuitability of fluid-attenuated inversion recovery vascular hyperintensity scores.

Fluid-attenuated inversion recovery (FLAIR) vascular hyperintensity (FVH) is used to assess leptomeningeal collateral circulation, but clinical outcomes of patients with FVH can be very different. The aim of the present study was to assess a FVH score and explore its relationship with clinical outcomes. Patients with acute ischemic stroke due to middle cerebral artery M1 occlusion underwent magnetic resonance imaging and were followed up at 10 days (National Institutes of Health Stroke Scale) and 90 days (modified Rankin Scale) to determine short-term clinical outcomes. Effective collateral circulation indirectly improved recovery of neurological function and short-term clinical outcome by extending the size of the pial penumbra and reducing infarct lesions. FVH score showed no correlation with 90-day functional clinical outcome and was not sufficient as an independent predictor of short-term clinical outcome.

MRI detects protective effects of DAPT treatment with modulation of microglia/macrophages at subacute and chronic stages following cerebral ischemia.

Notch homolog 1 (Notch 1) signaling is regarded as a potential therapeutic target for modulating the inflammatory response and exhibiting neuroprotective effects in cerebral injury following stroke. N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-phenylglycine t­butylester (DAPT) efficiently inhibits activation of the Notch 1 signaling pathway in microglia and may protect brain tissue from ischemic damage. However, the temporal proliferation and morphological alterations of microglia/macrophages throughout progression of the disease, as well as the comprehensive alterations of the whole brain following DAPT treatment, remain to be elucidated. The present study evaluated the temporal proliferation and the morphological alterations of microglia/macrophages over the period of the subacute and chronic stages, in addition to dynamic alterations of brain tissue, using the magnetic resonance imaging (MRI) method, following DAPT treatment. Sprague­Dawley rats (n=40) were subjected to 90 min of middle cerebral artery occlusion and were treated with DAPT (n=20) or acted as controls with no treatment (n=20). The two groups of rats underwent MRI scans prior to the induction of stroke symptoms and at 24 h, 7, 14, 21 and 28 days following the stroke. A total of five rats from each group were sacrificed at 7, 14, 21 and 28 days following induction of stroke. Compared with control rats, the MRI data of the ipsilateral striatum in treated rats revealed ameliorated brain edema at the subacute stage and recovered brain tissue at the chronic stage. In addition to this, treatment attenuated the round­shape and promoted a ramified­shape of microglia/macrophages. The present study confirmed the protective effect of DAPT treatment by dynamically monitoring the cerebral alterations and indicated the possibility of DAPT treatment to alter microglial characteristics to induce a protective effect, via inhibition of the Notch signaling pathway.

Evaluation of the characteristics of hepatic focal nodular hyperplasia: correlation between dynamic contrast-enhanced multislice computed tomography and pathological findings.

OBJECTIVE: To evaluate the characteristics of enhancement of focal nodular hyperplasia (FNH) of the liver by analyzing the dynamic contrast-enhanced multislice computed tomography (MSCT) features and correlating them with pathological findings. PATIENTS AND METHODS: Nine males and 16 females with pathologically confirmed FNH and complete preoperative contrast-enhanced MSCT data were recruited for this study. The imaging features of FNH on the pre- and postcontrast MSCT were analyzed by two experienced radiologists by consensus. RESULTS: Pathology showed central scars and abnormal blood vessels in 17 and 21 of 25 lesions, respectively, while MSCT with multiphase enhancement showed central scars in eight of the 17 lesions (47.1%) and abnormal arteries or draining veins in 13 of the 21 lesions (61.9%). Furthermore, abnormal draining veins in five lesions were found to be diagnostic, which is another important finding. CONCLUSION: Multiphase scanning can provide the panorama of FNH lesions and reveal their enhancement patterns and pathological characteristics. Abnormal blood vessels within or around the lesion are demonstrated more often than central scar, and both should be observed for FNH diagnosis.

The Initial Exploration of Adamkiewicz Artery Computed Tomographic Angiography With Monochromatic Reconstruction of Gemstone Spectral Imaging.

OBJECTIVE: The study aimed to explore whether optimal monochromatic reconstruction can improve the depiction of the Adamkiewicz artery (AKA) on gemstone spectral computed tomographic angiography (GSCTA) compared with the polychromatic reconstruction protocol. METHODS: The prospective study was approved by the ethics committee, and written informed consent was obtained from each patient. The 58 consecutive patients suspected of aortic aneurysm or dissection underwent aortic GSCTA. All images were reconstructed with both polychromatic (group A) and optimal monochromatic (group B) protocol. The CT values of the descending aorta and muscle, background noise, and the contrast-to-noise ratio were measured and calculated. With the criterion standard display of AKA, characteristic hairpin curve sign, 2 blinded radiologists analyzed data independently with the paired samples t, χ, and Mann-Whitney U test. RESULTS: The CT value of the descending aorta and the contrast-to-noise ratio of group B were significantly superior to group A (t = 12.7, P < 0.01; t = 15.2, P < 0.01). The visual rate of AKA (94.8%) in group B was significantly higher (χ = 4.2, P = 0.04) than group A (82.8%). Using a 5-point scale to assess, the score of the visualization efficiency of group B (226) was significantly higher (Z = -2.4, P = 0.02) than group A (192). CONCLUSIONS: The optimal monochromatic reconstruction for GSCTA can improve the visualization efficiency and quality of the AKA compared with the polychromatic reconstruction protocol.

Molecular imaging of angiogenesis to delineate the tumor margins in glioma rat model with endoglin-targeted paramagnetic liposomes using 3T MRI.

PURPOSE: To evaluate the use of endoglin-targeted paramagnetic liposomes in delineating the glioma margins using magnetic resonance (MR) angiogenesis imaging in a rat model. MATERIALS AND METHODS: Four liposome preparations, including nontargeted paramagnetic liposomes (Gd-SLs), isotype control IgG-coupled paramagnetic liposomes (IgG-Gd-SLs), endoglin monoclonal antibody coupled paramagnetic liposomes (MAb-Gd-SLs), and biotinylated antibodies (Bio-MAb)/streptavidin-coupled paramagnetic liposomes (SAv-Gd-SLs) for two-step pretargeting imaging, were formulated. All animal experiments were carried out with the approval of the Shanghai Animal Care. C6 glioma-bearing Sprague-Dawley rats were intravenously injected with gadolinium-diethylenetriaminepentaacetic acid (Gd-DTPA) or the previously mentioned liposomes (n = 5) and imaged with MR. T1 -weighted MRI was performed before and dynamically repeated after different contrast agents were injected. The enhancement features of the tumors were compared. RESULTS: The signal enhancement of the tumor in the two-step pretargeting group increased by 117.9 ± 5.3% at the periphery and 109.2 ± 3.5% in the center (P = 0.032) at the 8-hour timepoint after SAv-Gd-SLs injection. Ring-like enhancement margins were demonstrated at the periphery of the tumor in the two-step targeted group. The specificity of the targeted liposomes was supported by the competitive study. The signal of peak enhancement using MAb-Gd-SLs was 59% less than that of the two-step group and only slightly higher than the non-targeted groups. CONCLUSION: The two-step endoglin-targeted imaging using biotin-streptavidin interaction was demonstrated to induce intense enhancement of the tumor periphery, which implies that this advanced MR molecular contrast agent may be suitable for accurately delineating glioma tumor margins. J. Magn. Reson. Imaging 2015;41:1056-1064. © 2014 Wiley Periodicals, Inc.

The expression patterns of Reg IV gene in normal rat reproduction system.

Reg IV, the latest member of the regenerating gene family, has been documented in different tissues of human and rat, such as the colon, small intestine, stomach, and pancreas. Expression of Reg IV gene in distinct cell types has been correlated with its various functions in regeneration, cell growth and survival, proliferation and differentiation, cell adhesion, and resistance to apoptosis. However, there was no evidence to show whether the Reg IV protein is present in the reproductive system of normal rat. The aim of this study was to reveal the expression patterns of Reg IV in rat ovary and uterus. The expression of Reg IV was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot at mRNA and protein levels, respectively. The localization of Reg IV protein within rat ovary and uterus was investigated by immunohistochemistry (IHC). Our results showed that the expression of Reg IV in ovary was significantly higher than that in the uterus. The strong immunoreactive signals of Reg IV was observed in granulosa cells and oocytes of ovarian follicles, corpus luteum, and interstitial cells in rat ovary; only weak signals were detected in luminal and gland epithelium of rat endometrium. These findings first demonstrate the expression of Reg IV in ovary and uterus of the healthy rat at both mRNA and protein levels. It provides an evidence of Reg IV expression in rat reproductive system, which may help elucidate a potential role in cell growth and proliferation of reproductive system.

[Effects and mechanisms of autophagy of trophoblast cells in severe preeclampsia].

AIM: To investigate the occurrence law of autophagy in trophoblast cells from severe preeclampsia and its possible mechanisms of wrecking cells. METHODS: 23 cases of placenta tissues were collected from women suffer from severe preeclampsia and normal pregnant women respectively. Surface villous of placental was observed by scanning electron microscope, and ultrastructure and autophagosome of trophoblast cells were observed by transmission electron microscopic. The expressions of Beclin-1 and LC3II in placenta tissues were detected by Immunohistochemical technique, then its relevance whit weight of fetus and placental was analyzed. RESULTS: Compared with the control group, villous were decrease in placenta tissue from patients suffer from severe preeclampsia, and its arrangement was disordered; typical autophagosomes of trophoblast cells were observed by transmission electron microscope. The expression of Beclin-1, LC3IIwere significantly up-regulated in placenta tissue from patients suffer from severe preeclampsia.The correlation coefficient of Beclin-1, LC3II and placental weight, fetal weight respectively were -0.977, -0.930, -0.812, -0.849. CONCLUSION: Autophagy were significantly up-regulated in trophoblast cells from patients suffer from severe preeclampsia by increasing expression of Beclin-1, LC3II protein, and its expression was negatively related to weight of fetus and placental.

[Effects and molecular mechanisms of inhibiting FOXM1 expression on ovarian cancer cell line SKOV3 by Lentiviral vector].

AIM: To investigate the effects and possible molecular mechanisms of inhibiting FOXM1 expression on SKOV3 cells by lentiviral vector targeting FOXM1 shRNA. METHODS: SKOV3 cells were infected by lentiviral vector targeting FOXM1 shRNA with a multiplicity of infection (MOI) of 20, then growth curve of SKOV3 cells was determined by MTT assay, cell cycle was analysed by flow cytometry(FCM), and the expression of mRNA and protein of FOXM1, Cyclin D1, PLK1 by Real time PCR and Western blot. RESULTS: Lentiviral vector targeting FOXM1 shRNA with a multiplicity of infection (MOI) of 20 could significantly inhibit the growth of SKOV3 cells. After infected by lentiviral vector targeting FOXM1 shRNA, the G(0);/G(1); phase cells increased and the S-phase cells decreased, and the expression of mRNA and protein of FOXM1, Cyclin D1, PLK1 of SKOV3 cells were significantly down-regulated. CONCLUSION: Inhibiting FOXM1 expression has a significantly effect of inhibiting proliferation on SKOV3 cells. Blocking SKOV3 cells in the G(0);/G(1); phase by down-regulating the expression of Cyclin D1, PLK1 protein may be its mechanism.

Gadd45α as an upstream signaling molecule of p38 MAPK triggers oxidative stress-induced sFlt-1 and sEng upregulation in preeclampsia.

Preeclampsia (PE) is known to be associated with increased circulating levels of anti-angiogenic factors, such as soluble fms-related tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng). However, the way that placental oxidative stress results in the elevation of these two factors remains enigmatic. We have observed the overexpression of growth arrest and DNA damage-inducible 45 alpha (Gadd45α) and excessive activation of p38 mitogen-activated protein kinase (MAPK) in preeclamptic placentas compared with normotensive controls, together with increased levels of sFlt-1 and sEng in maternal sera in patients with PE. Moreover, Gadd45α knockdown or p38 inhibition provides protective effects in hypoxia/reoxygenation (H/R)-exposed human umbilical vein endothelial cells (HUVECs) by suppressing oxidative stress, inhibiting apoptosis, and promoting their potential for in vitro angiogenesis. A regulatory signaling pathway in which H/R intervention causes the induction of Gadd45α leading to p38 activation and ultimately an increase in sFlt-1 and sEng secretion in HUVECs has concurrently been established. Our study opens up a promising new avenue of investigation for increasing the understanding of the origin of sFlt-1 and sEng in PE and provides novel therapeutic targets for pregnancy complications arising from placental endothelial dysfunction.

[Study on p38 mitogen activated protein kinase in vascular endothelial cells dysfunction in preeclampsia].

OBJECTIVE: To study expression and activation of p38 mitogen activated protein kinase (MAPK) in vascular endothelial cells dysfunction in preeclampsia. METHODS: From Sept. 2009 to Mar. 2010, 54 pregnant women underwent deliveries in the First Affiliated Hospital of Chongqing Medical University were enrolled in this study, including 20 patients in mild preeclampsia group, 16 patients in severe preeclampsia group and 18 women with term cesarean section without perinatal complications as control group. Placental endothelial cells were labeled by CD34 to assay microvessel density (MVD) of each group. Immunohistochemical SP and western blot were used to detect localization and expression of p-p38 MAPK protein, respectively. The levels of sera soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng) were measured by ELISA. RESULTS: (1) The MVD of placenta were 103 ± 3 in control group, 81 ± 5 in mild preeclampsia group and 63 ± 4 in severe group, respectively, which showed statistical difference among each group (P < 0.05). (2) The expression of p38 MAPK protein were 0.84 ± 0.05 in control group, 0.90 ± 0.14 in mild group and 0.86 ± 0.18 in severe group, which did not reach remarkable difference among each group (P > 0.05). The expression of p-p38 MAPK protein were 0.13 ± 0.05 in control group, 0.59 ± 0.12 in mild group and 1.16 ± 0.18 in severe group, which show statistical difference among each group (P < 0.05). (3) The localization of p-p38 was in trophoblast, endothelial cells and a few stromal cells in placenta. (4) The level of sFlt-1 and sEng: (1) The concentration of sFlt-1 and sEng were (5.2 ± 0.3) and (10.9 ± 0.4) µg/L in control group, (12.5 ± 1.2) and (20.4 ± 5.3) µg/L in mild group and (19.3 ± 3.0) and (29.5 ± 3.7) µg/L in severe group. When drawing paired comparison in those groups, the differences showed statistical difference (P < 0.05). (2) There were positive correlations between p-p38 MAPK protein levels and the concentrations of serum sFlt-1, sEng in preeclampsia groups (r = 0.68, P < 0.05; r = 0.87, P < 0.05). CONCLUSIONS: The remarkable activation of the p38 MAPK in the placenta of patients with preeclampsia induced the increased levels of sFlt-l and sEng in maternal serum, which confer the injury of vascular endothelial cells that caused the significant decline of MVD in placentas. p38 MAPK signaling might be one of the key pathways in vascular endothelial cell dysfunction in preeclampsia.

[Report of a case with Joubert syndrome and literature review].

OBJECTIVE: To explore the clinical feature, imaging and their diagnostic value for Joubert syndrome (JS). METHOD: The clinical data, imaging feature, and 31 references from China Biomedical literature database (CBMdise) were reviewed and analyzed. RESULT: The age of onset of 32 patients including male 20 and female 12 ranged from 3 days to 6 years (mean 2.2 years). All the 32 patients with Joubert syndrome showed "slow growth" and "reduced muscle tension", 26 cases (81.3%) showed "gasp for breath", 26 cases (81.3%) showed "unusual motion of eyeball", 2 cases (6.3%) showed additional fingers (toes), 6 cases (18.8%) showed stretching tongue with agape. The typical imaging features of Joubert syndrome included "molar tooth sign", "midline cleavage" between cerebellar hemispheres and "bat-wing" like fourth ventricle, all the 32 patients with Joubert syndrome showed "midline cleavage", "molar tooth sign" was present in 29 cases (90.1%), and "bat-wing" like fourth ventricle in 30 cases (93.8%). CONCLUSION: Joubert syndrome is a rare congenital brain malformation. The typical clinical manifestations included "gasp for breath", "reduced tension of muscle", "slow growth" and "unusual motion of eyeball", and at the same time the patients had the following typical imaging features of brain: "molar tooth sign", "midline cleavage" and "bat-wing" like fourth ventricle.

CT volume measurement for prognostic evaluation of unresectable hepatocellular carcinoma after TACE.

AIM: To analyze the value of computed tomography (CT) volume measurements for evaluation of the survival rate of unresectable hepatocellular carcinoma (HCC) patients after transcatheter arterial chemoembolization (TACE). METHODS: One hundred and sixty-six unresectable HCC patients after TACE were involved in this retrospective study. Hepatic CT scan was performed for all patients before and 4 wk to 2 mo after TACE to define the morphologic features of HCC including its largest diameter, volume, product of the greatest axial dimension, tumor to liver volume ratio (TTLVR), and tumor shrinkage ratio. Clinical variables used in the study included gender, age, pattern of tumor growth, number of lesions, Child-Pugh classification of liver function, repeated TACE times, pre- or post-treatment alpha-fetoprotein (AFP) level, portal vein cancerous thrombus, tumor metastasis, degree of lipiodol retention within the tumor, and percutaneous ethanol injection. The correlation between survival time and clinical variables of patients or lesions was analyzed by combining morphologic features with the corresponding clinical and general data as input. A Cox proportional hazard model was used to analyze prognostic factors. The Kaplan-Meier method was used to calculate the cumulative survival time. Influence of the parameters on prognosis was analyzed by the log-rank test. RESULTS: The overall 6, 12, 24, 36 and 60 mo cumulative survival rates were 78.92%, 49.85%, 23.82%, 15.60% and 8.92%, respectively. The median survival time was 12 mo. Univariate and multivariate analysis showed that only 4 parameters were the independent prognostic factors including TTLVR (chi(2) = 14.328, P < 0.001), portal vein cancerous thrombus (chi(2) = 5.643, P = 0.018), repeated TACE times (chi(2) = 8.746, P = 0.003), and post-treatment serum AFP level (chi(2) = 5.416, P = 0.020). When the TTLVR value was less than 70%, the survival time was inversely correlated with the TTLVR value. CONCLUSION: CT volume measurement technique can predict the prognosis of unresectable HCC patients after TACE.

The role of Crk/Dock180/Rac1 pathway in the malignant behavior of human ovarian cancer cell SKOV3.

Small GTPases, particularly the Rho family, are key regulators of cell motility and migration. Dock180 was well known for the main target of signal adaptor protein Crk and acted as a guanine-nucleotide exchange factor for small GTPase Rac1. In the present study, Dock180 was found to combine primarily with CrkI other than CrkII, and its association with Elmo1 was also demonstrated in ovarian cancer cell SKOV3. To evaluate the role of Dock180 in human ovarian cancer cell, we performed RNAi-mediated knockdown of Dock180 in SKOV3 cells using small interfering RNA expression vector. In Dock180 knockdown cells, we found that Elmo1 expression and Rac1 activity were decreased simultaneously. By contrast, the expressions of both another Crk-combining molecule C3G and Rap1 activity were observed to increase obviously. Accordingly, all Dock180 knockdown cells present with evident change in cell morphology, reduced cell proliferation, and attenuated cell migration. Taken together, these results suggest that signal transfer of Crk/Dock180/Rac1 is implicated in actin cytoskeleton reorganization and thus in the cell proliferation, motility, invasion, and of human ovarian cancer cell line SKOV3.

[Polymorphism of estrogen receptor alpha genes Pvu I, Xba I and TA repeat sequence and hypomenorrhea with unknown aetiology].

OBJECTIVE: To study the roles of the polymorphism of the estrogen receptor genes in hypomenorrhea with unknown aetiology. METHODS: A case control study was carried out in south west of China, with 100 patients with hypomenorrhea in the case group and 100 eumenorrhea women in the control group. Molecular biology test was undertaken to test the restriction fragment length polymorphism (RFLP) of the first intron incision enzyme Pvu II, Xba I in ERa gene. Depuration, clone and sequence analysis was performed to the TA repeated sequence in the hypervariable region of estrogen receptor gene. The genotype distribution of ERa gene polymorphism was compard between the case and control groups. RESULTS: The patients with hypomennorrhea had higher P genotypic frequency (47.5%) than the control (30.5%), with an OR of 1. 810 (95% CI = 1.113-2.765, P = 0.012). The patients with hypomennorrhea had lower X genotypic frequency (20.5%) than the control (32.0%), with an OR of 0.641 (95% CI = 0.361-0. 898, P = 0.036). The patients with hypomennorrhea had higher frequency of TA13 allele (P = 0.006) and lower Frequency of TA15 allele frequency (P = 0.033) than the control. CONCLUSION: ERa gene polymorphism is associated with hypomenorrhea with unknown aetiology. P allele and TA13 allele may be risk factors, while X allele and TA15 allele may be protective factors.

Magnetic resonance angiography of carotid and cerebral arterial occlusion in rats using a clinical scanner.

In rat models to induce both focal cerebral ischemia and chronic cerebral hypoperfusion, it is highly desirable to verify the success of vessel occlusion and reopening with non-invasive method. The contrast-agent free 3D time-of-flight magnetic resonance angiography (TOF-MRA), diffusion-weighted imaging (DWI) and T2-weighted imaging by 3.0-T MR clinical scanner were applied when unilateral middle cerebral artery (MCA) was occluded and reopened, and after bilateral common carotid arteries were in ligation. The arterial angiograms of the rat brain and neck were achieved successfully in all chosen directions by the 3D TOF-MRA. It was shown that MCA in occlusion presented no signal in MRA, and the parenchyma of the ipsilateral MCA territory hypointensity signal in maps of apparent diffusion coefficient (ADC). After reperfusion, the signal intensity of ipsilateral MCA was resumed in MRA, and the decreased ADC was restored simultaneously. However, after 5h of reperfusion, it was found that the value of ADC deteriorated second time with high T2 value. In bilateral common carotid artery occlusion (BCCAO) rats, it can be confirmed by MRA that the effectively occluded BCCA presented the absent signal and the basilar artery became tortuous. As a result, MRA by clinical scanner was proved of a valuable method to validate transient middle cerebral artery occlusion (MCAO) and permanent BCCAO rat model.

[Association between Val80 polymorphism of the CYP19 gene, A163G polymorphism of the OPG gene and bone mineral density in post-menopausal Chinese women].

We investigated the association of the G/A polymorphism at Val80 of the cytochrome P450 family 19 (CYP19) gene, the A163G polymorphism of the osteoprotegerin (OPG) gene with bone mineral density (BMD) in 200 randomly selected postmenopausal women in Chongqing. Single nucleotide polymorphisms were detected by polymerase chain reac-tion-restriction fragment length polymorphism (PCR-RFLP). BMD of the proximal femur and lumbar spine (L2-4) was measured by NORLAND XR-46 dual-energy X-ray absorptiometer (DEXA). The frequencies of genotypes in these women were as follows: GG (19.5), GA (44.5%), AA (36.0%) for the Val80 polymorphism in CYP19; and AA (13.0%), AG (42.0%), GG (45.0%) for the A163G polymorphism in OPG. The distribution of genotype frequency was in Hardy-Weinberg equilibrium (P0.05). ANCOVA and multiple stepwise regression analysis showed the Val80 polymor-phism in the third exon of the CYP19 gene was not associated with the BMD in postmenopausal women (P0.05). Except for the trochanter region, BMD at the femoral neck, Ward's triangle, and L2-4 was lower in subjects with AG/GG/AG+GG genotypes than those with the AA genotype for the A163G polymorphism and A163G genotypes were associated with BMD at these skeletal regions in postmenopausal women (P0.05). A163G polymorphism resides in the promoter region of the OPG gene and its genotype distribution is significantly different among different ethnic groups. Our results indicate that the AA genotype might have some beneficial effect on BMD and the variant G allele might reduce BMD in postmenopausal women.