[Predictive value of plasma exosomal miR-124-3p for the risk of chronic cerebral hypoperfusion].

Objective: To investigate the predictive value of plasma exosomal microRNA (miR)-124-3p in the risk of chronic cerebral hypoperfusion (CCH). Methods: A case-control study. Thirty patients who were diagnosed with CCH (CCH group) based on cranial artery spin labeling (ASL) in the neurology outpatient clinic of Sichuan Provincial People's Hospital from March 2022 to June 2022 and 30 healthy volunteers (control group) were included. Age, gender, smoking history, alcohol consumption history, diabetes history, hypertension, hyperlipidemia history, uric acid, fasting blood glucose, homocysteine and plasma exosomal miR-124-3p expression level were compared between the two groups. Comparisons of categorical variables were analyzed by either χ2 test or Fisher's exact test. If the data of continuous variables followed a normal distribution, they were expressed as mean±standard deviation (SD) and compared by t-test for two independent samples; otherwise, the data were expressed as M(Q1, Q3), and analyzed by Mann-Whitney U test for comparison between two groups. The correlation between cerebral blood flow and exosomal miR-124-3p levels was analyzed by Pearson's correlation. Binary multifactorial logistic regression analysis was used to determine the risk factors associated with CCH, and corresponding odds ratios (OR) and 95% confidence intervals (CI) were calculated. P<0.05 was considered significant. Results: There was no significant difference in age (64±8 vs. 60±8 years old), gender (33.3% vs. 30.0%), history of smoking (20.0% vs. 3.3%), alcohol consumption (20.0% vs. 6.7%), diabetes mellitus (13.3% vs. 13.3%), hypertension (53.3% vs. 30.0%), history of hyperlipidemia (46.7% vs. 36.7%), uric acid (288±60 vs.319±67 µmol/L), and fasting glucose [4.99(4.63, 5.91) vs. 5.28(5.09, 6.05) mmol/L] and homocysteine [11.35(10.18, 13.08) vs.11.00(9.78, 13.03) µmol/L] between the CCH and control groups (P>0.05). Plasma exosomal miR-124-3p expression was significantly higher in the CCH group than in the control group [13.08 (8.59, 21.55) vs. 2.85 (1.44, 5.10), respectively; U=169.50, P<0.001]. Pearson's correlation test showed that the level of exosomal miR-124-3p was negatively correlated with cerebral blood flow in the hypoperfused region in patients with CCH (r=-0.932, P<0.001). Multi-factor logistic regression analysis showed that plasma exosomal miR-124-3p was independently associated with the risk of CCH (OR=1.169,95%CI 1.063-1.286,P=0.001). Conclusions: The expression of plasma exosomal miR-124-3p is negatively correlated with cerebral blood flow in areas of low perfusion and is an independent risk factor for CCH. Plasma exosomal miR-124-3p may thus serve as a valid biomarker for CCH risk prediction.

[Extranodal nasal type natural killer/T-cell lymphoma of the digestive system: a clinicopathological study of thirteen cases].

Objective: To investigate the clinicopathological features, and diagnostic and differential diagnostic characteristics of extranodal nasal type natural killer/T-cell lymphoma (ENKTCL) of the digestive system. Methods: Thirteen cases of ENKTCL in the digestive system were collected at the Henan Provincial People's Hospital, Zhengzhou, China, from August 2000 to August 2020. The histopathological, immunohistochemical and in situ hybridization features were analyzed, as well as those of T-cell receptor (TCR) gene rearrangement in some cases. The patients were followed up. Results: There were 11 males and 2 females. The age ranged from 28 to 80 years (median=53 years). Seven cases were present in the colorectum, and 3 cases were present in the small intestine. The other three cases were in stomach, gallbladder and liver (one case each). The main clinical symptoms were fever, and abdominal pain, often accompanied by fatigue, diarrhea, hematochezia, elevated serum albumin, elevated lactate dehydrogenase, and increased peripheral blood EB virus DNA copy. Histologically, the tumor accompanied by a heavy admixture of inflammatory cells (small lymphocytes, plasma cells and histiocytes). There was diffuse dense tumor cell infiltrate, with prominent coagulative necrosis. The lymphomatous infiltrate had angiocentric and angio-necrotic changes. Immunohistochemically, lymphoid cells expressed CD3 in all cases. Some of them showed weakened/absent other T cell markers, while all of them expressed CD56 except 1 case. A few of the cases showed CD4-/CD8+ killer T cell phenotypes. In situ hybridization showed EB virus encoded RNA (EBER) was positive in all cases. Clonal TCR gene rearrangement was not detected in all 7 cases tested. The median survival time was 9 months. Conclusions: ENKTCL of the digestive system is extremely rare. It often predisposes the patients to acute abdomen such as perforation of the gastrointestinal tract. The treatment outcomes are dismal, and the prognosis is poor. Clinical and imaging studies are often non-specific. It is also easy to be misdiagnosed as non-specific ulcers. Combined with immunohistochemistry, in situ hybridization and TCR gene rearrangement analysis and better understanding of this tumor's clinicopathological characteristics can help improve its diagnosis and early treatment.

[Clinicopathological features of verrucous hemangioma].

Objective: To investigate the clinicopathological features, and differential diagnosis of verrucous hemangioma (VH). Methods: Twenty-eight VH cases diagnosed from 2005 to 2020 in Henan Provincial People's Hospital, Zhengzhou, China were analyzed retrospectively. Immunohistochemical studies were used to detect diagnostic markers. The mutation status of PIK3CA (exons 9 and 20) was detected using fluorescence PCR. Results: There were 13 males and 15 females in 28 cases, with the male to female ratio of 1.0∶1.2. There were 25 patients under the age of 18 years. The age range was from 10 months to 56 years (mean, 9.7 years; median, 4.5 years). There were 17 cases occurred in the lower extremities, 7 in the upper extremities and 4 in the trunk. All 28 cases were irregular red patches on the skin, which grew slowly. Some of them were thickened with uneven surface, which was light pink or red-white. Skin lesions of the 7 cases ranged from dark red and reddish brown, with a rough and hard surface. Satellite foci were present. Microscopically, 28 cases had a wide range of pathological features. Dilated, malformed vessels were observed from dermal papilla to deep soft tissue. Among them, the dermal papillary layer was mainly composed of many proliferating and expanding thin-walled capillaries and cavernous blood vessels. Thin-walled small vessels were found in the dermal reticular layer and subcutaneous fascia layer, with no obvious endothelial cell proliferation, occasional papillary hyperplasia, and lobular distribution of the malformed vessels in the fascia layer mixed with the fibroadipose tissue. There was epidermal papillary hyperplasia with hyperkeratosis and parakeratosis, lengthening and mutual fusion of epithelial horns. Immunohistochemistry showed that CD31, CD34, ERG and WT-1 were diffusely and strongly positive. The expression of GLUT-1 was present in superficial dermal vascular endothelial cells, but undetectable in the deep layer. The PIK3CA tests of 13 cases showed that no somatic mutations were found in exons 9 and 20. Twenty-five patients were followed up for 5 months to 10 years. Seven patients underwent multiple surgical resections and plastic surgeries due to the large size, and 8 patients had recurrence. Conclusions: VH is a rare congenital vascular malformation and more commonly occurs in infants and children. It tends to appear in limbs, especially lower limbs and distal limbs. Its morphology and immunophenotype are characteristic and should be distinguished from other vascular malformations and the resolution phase of infant hemangiomas. In about one third of the cases, postoperative recurrence may occur and long-term follow-up is often required.

Environmental Exposure-Associated Human Health Risk of Dioxin Compounds in the Vicinity of a Municipal Solid Waste Incinerator in Shanghai, China.

In order to assess environmental exposure-associated human health risk of dioxin compounds for the population in the vicinity of a municipal solid waste incinerator (MSWI) in Shanghai, the atmospheric samples (n = 24) and soils samples (n = 96) were collected and analyzed to obtain the concentration level, pollution characteristics and seasonal changes of dioxin compounds in environmental medias. The toxicity equivalent concentration range of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) was 30.9-409 fg WHO-TEQ·m-3 in atmosphere and 0.362-8.55 ng WHO-TEQ·kg-1 in soil. The non-carcinogenic health risk and carcinogenic health risk from PCDD/Fs environmental exposure of people living in the vicinity of the MSWI in Shanghai were all within the allowable range of the US Environmental Protection Agency, which implied that the MSWI in Shanghai did not produce additional risk for the population living in its vicinity.

[Application of pump-controlled retrograde trial off in weaning from veno-arterial extracorporeal membrane oxygenation in adult patients].

Objective: To Summarize the experience of pump-controlled retrograde trial off (PCRTO) in the process of veno-arterial extracorporeal membrane oxygenation (VA-ECMO) withdrawal in adult patients. Methods: Adult patients who received ECMO assistance in Intensive Care Unit for Cardiac Surgery from March to July 2019 were collected. According to our strategies, PCRTO was used if the patients can wean from VA-ECMO and hemodynamic indexes were recorded during the process. The statistics data was collected, including the 48 hours survival rate, ECMO re-assistance rate, thrombus complications, Intensive Care Unit (ICU) stay time and hospital stay time after weaning from VA-ECMO. The patients who failed in the test were continued to be assisted by ECMO. Results: There were 46 patients assisted by VA-ECMO in our center. In total, 21 adults who met the offline test standard underwent 26 PCRTOs, including 10 male adults (47.6%), with an age of 65 (55, 68) years old. Eighteen adults passed the withdrawal test. No new thrombus was found in the arteriovenous ultrasound of the lower extremity after weaning from ECMO, and no pulmonary embolism was found in the chest X-ray. The success rate of weaning from ECMO was 69.23%(18/26). The D-dimer decreased [584(348,2 107)µg/L vs 1 440(631,2 916)µg/L, P=0.014] and the left ventricular ejection fraction (LVEF) increased (51.4%±8.5% vs 46.9%±10.6%, P=0.013) on the next day after weaning. There were significant differences in heart rate (HR), central venous pressure (CVP), oxygenation index and lactate (Lac) during the PCRTO in the group which involved the cases of the 8 failed experiments (all P<0.05). Compared with the failure group, there were significant differences in age, blood flow rate, CVP before the test, HR, pulse oxygen saturation(SpO(2)), CVP, Lac and oxygenation index after the test, and the variations of SpO(2), CVP and Lac. Conclusion: PCRTO is a simple, reversible, safe and effective weaning method. It can be used in the process of VA-ECMO withdrawal in adult patients.

[A comparative study of 2- and 24-hour creatinine clearance in patients in intensive care unit].

Objective: To analyze the correlation and consistency of 2- and 24-hour endogenous creatinine clearance rate (Ccr) in patients in the intensive care unit (ICU). Methods: It was a prospective bicentric observational study.According to the inclusion criteria, the patients were selected from the ICU of Fuxing Hospital Affiliate to Capital University of Medical Sciences and Chuiyangliu Hospital Affiliated to Tsinghua University from September 2015 to December 2016.After inclusion, the mixed urine sample was collected at 9-11 am, 9-11 pm and all day long (7 am-7 am, 24 hours). The urine volume was recorded and the urine creatinine was detected.The correlation and consistency analysis of Ccr were applied in the group of 9-11 am, 9-11 pm and 7 am-7 am.Clinical data including sex, age, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ), the use of diuretics, mean arterial pressure, blood glucose and body temperature were recorded for the analysis of the factors affecting correlation and consistency of Ccr between 2 h and 24 h. Results: A total of 158 cases were included, 82(51.9%) cases were males, the mean age was (76±13) years, the APACHE Ⅱ score was 19±8.And 83 patients (52.5%) were admitted to the ICU due to pulmonary disease.The average Ccr in the group of 24 h was (70±58) ml/min, the Ccr in 105 patients (66.5%) was less than 80 ml/min, it was less than 60 ml/min in 88 patients (55.7%) and was less than 30 ml/min in 33 patients (20.9%). The fasting serum creatinine level in the second day morning after admitted to ICU was 77.0(52.8, 122.2) µmol/L.The correlation analysis showed that the relative indices of R(2) between the 24hCcr and 2hCcr in the morning (9-11 am) and evening (9-11 pm) was 0.704 and 0.825, respectively, both P<0.01, and the bias was -2.3 and 0.3 ml/min.The patients were divided into group A (D-value≤20%) and B (D-value>20%) according to the Ccr D-value of the 24 h and the 2 h in morning.There were 72 patients (45.6%) in group A, and 86 patients (54.4%) in group B. The multiple Logistic regression analysis indicated that the factors affecting the correlation and consistency between 2hCcr and 24hCcr were body temperature, age and the D-value of serum creatinine, and the odd ratio (95%CI) was 1.844(1.112-3.059), 1.046(1.020-1.072), 1.038(1.003-1.074), respectively, all P<0.05. Conclusions: There is significant correlation and consistency between 2hCcr and 24hCcr in ICU patients.And in most of the cases, the 24hCcr can be replaced by random 2hCcr, but the affecting factors of body temperature, age and the D-value of serum creatinine should be taken into account.

[Preventive effects of ulinastatin on acute respiratory distress syndrome].

OBJECTIVE: To explore the effect of ulinastatin on prevention of acute respiratory distress syndrome (ARDS). METHODS: A prospective multicentral cohort study was conducted. The patients from three intensive care units (ICUs) of grade A tertiary hospitals in Beijing and a ICU of grade A tertiary hospitals in Cangzhou from January 2012 to December 2014, included 77 ARDS at-risk patients with ulinastatin treatment and 108 ARDS at-risk patients without ulinastatin treatment (control) were eligible. Both groups received normal treatment; additionally, the intervention group received 600 000 units of ulinastatin via intravenous infusion for 5 days. The control group received the same amount of saline via intravenous infusion for 5 days. Venous blood human neutrophil elastase (HNE) and peptidase inhibitor 3 (PI3) levels were measured on days 1, 3, and 7, respectively. Other outcomes included acute physiology and chronic health evaluation scoring II (APACHE II), body temperature, respiratory rate, heart rate, mean arterial pressure, white blood cell counts, PaO2/FiO2, ARDS incident, mechanical ventilation time, ICU treatment and hospitalization duration, 28 days mortality. RESULTS: The PI3 levels showed no statistical difference on day 1, but significant differences on day 3 and day 7 between the two groups (P<0.01). HNE/PI3 ratio showed no statistical difference on day 1, but significant differences on day 3 and day 7 (P<0.05). PaO2/FiO2 was significantly higher in ulinastatin group on day 3 and day 7 (P<0.05). The incident rate for ulinastatin group was 15.58%, lower than that for the control group (33.33%), and the difference was statistically significant (P<0.05). The mechanical ventilation time and ICU treatment time in ulinastatin group was shorter than that in the control group, and the difference was statistically significant (P<0.05). There were no significant effects in other factors. CONCLUSION: Increased dose of ulinastatin can recover the balance of HNE and its antagonist, lower the HNE's damage to lungs, and further reduce the ARDS incident rate.

[Preventive effects of ulinastatin on acute respiratory distress syndrome].

OBJECTIVE: To explore the effect of ulinastatin on prevention of acute respiratory distress syndrome (ARDS). METHODS: A prospective multicentral cohort study was conducted. The patients from three intensive care units (ICUs) of grade A tertiary hospitals in Beijing and a ICU of grade A tertiary hospitals in Cangzhou from January 2012 to December 2014, included 77 ARDS at-risk patients with ulinastatin treatment and 108 ARDS at-risk patients without ulinastatin treatment (control) were eligible. Both groups received normal treatment; additionally, the intervention group received 600 000 units of ulinastatin via intravenous infusion for 5 days. The control group received the same amount of saline via intravenous infusion for 5 days. Venous blood human neutrophil elastase (HNE) and peptidase inhibitor 3 (PI3) levels were measured on days 1, 3, and 7, respectively. Other outcomes included acute physiology and chronic health evaluation scoring II (APACHE II), body temperature, respiratory rate, heart rate, mean arterial pressure, white blood cell counts, PaO2/FiO2, ARDS incident, mechanical ventilation time, ICU treatment and hospitalization duration, 28 days mortality. RESULTS: The PI3 levels showed no statistical difference on day 1, but significant differences on day 3 and day 7 between the two groups (P<0.01). HNE/PI3 ratio showed no statistical difference on day 1, but significant differences on day 3 and day 7 (P<0.05). PaO2/FiO2 was significantly higher in ulinastatin group on day 3 and day 7 (P<0.05). The incident rate for ulinastatin group was 15.58%, lower than that for the control group (33.33%), and the difference was statistically significant (P<0.05). The mechanical ventilation time and ICU treatment time in ulinastatin group was shorter than that in the control group, and the difference was statistically significant (P<0.05). There were no significant effects in other factors. CONCLUSION: Increased dose of ulinastatin can recover the balance of HNE and its antagonist, lower the HNE's damage to lungs, and further reduce the ARDS incident rate.

Combinatorial biochemical and chemical analyses of polychlorinated dibenzo-p-dioxins and dibenzofurans in agricultural soils from Chongming Island, Shanghai, China.

We compared polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/F) concentrations [expressed as toxic equivalent quantities (TEQs)] in agricultural soil samples from Chongming Island (Shanghai, China) determined using two analytical approaches, an enzyme-linked immunoassay (EIA) method and a high resolution gas chromatography-high resolution mass spectrometry (HRGC/HRMS) method. The PCDD/F concentrations in all 31 soil samples were at background levels (7.30-16.7 pg EIA-TEQ/g from the EIA analysis and 0.526-1.99 pg WHO-TEQ/g from the HRGC/HRMS analysis). Although, the EIA method overestimated the PCDD/F concentrations compared with the concentrations determined using the HRGC/HRMS method. The absence of false-negatives showed by the EIA analysis verified that this method is useful for preliminary sample screening (prior to HRGC/HRMS analysis) and the preliminary characterization of potentially contaminated sites.

Levels of PCDD/Fs in agricultural soils near two municipal waste incinerators in Shanghai, China.

A study was conducted on polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/F) in agricultural soils at 41 sites within a radius of 3 km from two municipal solid waste incinerators in Shanghai. The PCDD/F concentrations ranged from 71.32 to 3,881.44 pg g⁻¹ (0.64-61.15 pg I-TEQ g⁻¹). The highest PCDD/F concentrations were found approximately 1,000 m from the municipal solid waste incinerators. The PCDD/F homologue profiles of all soil samples were compared with the profiles from suspected PCDD/F sources by multivariate statistical analysis. The results showed that, the PCDD/F pollutions in some soil samples can be attributed to emissions from the municipal solid waste incinerators.

Hypersensitivity reactions to penicillins: studies in a group of patients with negative benzylpenicillin G skin test.

BACKGROUND: Although skin tests are usually employed to evaluate current penicillin allergy status, a negative result does not exclude hypersensitivity. There is a need for accurate in vitro tests to exclude hypersensitivity. A radioallergosorbent test (RAST) is a potentially good supplementary approach, but there is little information on the suitability of this method to diagnose penicillin hypersensitivity in subjects with a negative skin test to benzylpenicillin. METHODS: A total of 133 patients with a negative skin test to benzylpenicillin G (PG) and all of whom developed allergic reactions to PG were studied. RAST was used to detect eight kinds of specific IgE antibodies to penicillins in serum, which included four kinds of major and minor antigenic determinants to four penicillin drugs. The combination sites for the specific IgE antibodies were studied by RAST inhibition test. RESULTS: The rate of positive reactions for the specific IgE antibodies was 59.40% (79/133). Of the eight kinds of antigenic determinants, the positive rates for specific IgE against the major and minor determinants were 39.10% (52) and 42.86% (57) respectively. Of the four drugs, positive cases only to PG were 10 (7.5%), were significantly fewer than the cross-reacting positive cases (36) to PG (P < 0.01). In the RAST inhibition studies all drugs exhibited good inhibitory potencies, and in some instances the side-chain of the penicillins could induce specific responses with a variable degree of cross-reactivity among the different penicillins. CONCLUSION: Radioallergosorbent test is a good complementary test in persons who are skin-test negative with PG, and the sensitivity of RAST increases with increasing specificity of IgE antibodies to be detected. 6-APA and the groups, making part of the different side-chains on penicillins, all contributed to the cross-reactivity.

Molecular Detection of Theileria species in Cattle from Jilin Province, China.

Bovine theileriosis is a tick-borne disease that is hampering the development of the domestic cattle industry in northern China. This study involved a molecular survey of bovine Theileria species in 137 blood samples from cattle in the Jilin province of China. The DNA samples were screened by species-specific 18S rRNA PCR. Results revealed that 19.7% (27/137), 17.5% (24/137) and 10.9% (15/137) were found to be infected with Theileria sinensis, Theileria orientalis, respectively. Mixed infection was found in 8.8% (12/137). The overall detection rates of Baishan, Yanji, Jilin and Liaoyuan districts was 60.0%, 17.5%, 5.3% and 0%, respectively. There is little information on the detection and distribution of bovine Theileria species in northern China. Therefore, this study provides important data for understanding the epidemiology of Theileria species and designing appropriate approaches for the diagnosis and control of bovine theileriosis in northern China.

Reassortment and modification of hemagglutinin cleavage motif of avian/WSN influenza viruses generated by reverse genetics that correlate with attenuation.

Avian influenza associated with H9N2 and H5N1 subtypes of avian influenza viruses (AIVs) has raised great concerns in China. To study this problem, reverse genetics has been employed. Three reassortants, rgH9N2, rgH5N1 and rgH5N2, were prepared and compared. Their hemagglutinin (HA) and neuraminidase (NA) genes originated from Chinese AIV isolates of H9N2 or H5N1 subtype, while the rest of their genes were derived from A/WSN/33(H1N1) virus (WSN). In the H5 HA reassortants, the multibasic cleavage site was converted to a monobasic one. The results demonstrated that the reassortants did not produce CPE on MDCK cells in the absence of trypsin, showed egg-adaptation phenotype and stability of HA and NA during consecutive egg passages, and were not lethal to chickens and mice. However, the rgH5N1 reassortant exhibited a residual virulence in terms of lethality to chick embryos and pathogenesis in chickens. It can be concluded that (i) the genetic modification of H5 HA attenuated the H5 reassortants, (ii) the presence of internal WSN proteins contributed to the attenuated properties of the reassortants independently on H5 HA, and (iii) also the overall genome composition contributed to virulence differences. This report provides further contribution of reverse genetics to the knowledge of virulence of influenza viruses.

Beta-arrestin1 and 2 differently modulate metabotropic glutamate receptor 7 signaling in rat developmental sevoflurane-induced neuronal apoptosis.

Beta-arrestins (ß-arrs) are initially known as negative regulators of G protein-coupled receptors (GPCRs). Recently, there is increasing evidence suggesting that ß-arrs also serve as scaffolds and adapters that mediate distinct intracellular signal transduction initiated by GPCR activation. In the previous study, we have shown that metabotropic glutamate receptor 7 (mGluR7) and extracellular signal-regulated kinase 1 and 2 (ERK1/2) signaling may be involved in the developmental sevoflurane neurotoxicity. In the present study, we showed that activation of mGluR7 with a group III mGluRs orthosteric agonist LAP4 or an atypical mGluR7 allosteric agonist N,N'-bis(diphenylmethyl)-1,2-ethanediamine dihydrochloride (AMN082) significantly attenuated sevoflurane-induced neuronal apoptosis. Interestingly, this neuroprotective role of LAP4 could be partially reduced by ß-arr1 small interfering RNA (siRNA) or ß-arr2 siRNA transfection. In contrast, ß-arr2 siRNA transfection alone abolished the effects of AMN082 on sevoflurane neurotoxicity. In addition, administration of LAP4 or AMN082 significantly enhanced Phospho-ERK1/2 in sevoflurane neurotoxicity, which could be abrogated by ß-arr2 siRNA transfection, but not by ß-arr1 siRNA transfection. Increased ß-arr2-dependent Phospho-ERK1/2 signaling alleviated sevoflurane neurotoxicity by inhibiting bad phosphorylation. We also found that the neuroprotective role of AMN082 was completely reversed by ERK1/2 inhibitor 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene (U0126). Alternatively, treatment with U0126 partially suppressed the neuroprotective of LAP4, suggesting that other mechanisms may be implicated in this process. Further investigation indicated that, in the scenario of sevoflurane neurotoxicity, application of LAP4 (but not AMN082) increased the interaction of ß-arrs with transcriptional factors CREB binding protein (CBP) and p300. LAP4 also enhanced the ß-arr1-dependent H3 and H4 acetylation in sevoflurane neurotoxicity. For the behavior study, treatment with LAP4 or AMN082 significantly improved the emotional and spatial learning and memory disorders induced by postnatal sevoflurane exposure. These results suggested that ß-arr1 and 2 may differently modulate mGluR7 signaling in developmental sevoflurane neurotoxicity. This study also reveals a ß-arr-biased agonism at GPCRs (e.g. mGluR7).

Suppression of tumor angiogenesis by targeting the protein neddylation pathway.

Inhibition of protein neddylation, particularly cullin neddylation, has emerged as a promising anticancer strategy, as evidenced by the antitumor activity in preclinical studies of the Nedd8-activating enzyme (NAE) inhibitor MLN4924. This small molecule can block the protein neddylation pathway and is now in clinical trials. We and others have previously shown that the antitumor activity of MLN4924 is mediated by its ability to induce apoptosis, autophagy and senescence in a cell context-dependent manner. However, whether MLN4924 has any effect on tumor angiogenesis remains unexplored. Here we report that MLN4924 inhibits angiogenesis in various in vitro and in vivo models, leading to the suppression of tumor growth and metastasis in highly malignant pancreatic cancer, indicating that blockage of angiogenesis is yet another mechanism contributing to its antitumor activity. At the molecular level, MLN4924 inhibits Cullin-RING E3 ligases (CRLs) by cullin deneddylation, causing accumulation of RhoA at an early stage to impair angiogenic activity of vascular endothelial cells and subsequently DNA damage response, cell cycle arrest and apoptosis due to accumulation of other tumor-suppressive substrates of CRLs. Furthermore, we showed that inactivation of CRLs, via small interfering RNA (siRNA) silencing of its essential subunit ROC1/RBX1, recapitulates the antiangiogenic effect of MLN4924. Taken together, our study demonstrates a previously unrecognized role of neddylation in the regulation of tumor angiogenesis using both pharmaceutical and genetic approaches, and provides proof of concept evidence for future development of neddylation inhibitors (such as MLN4924) as a novel class of antiangiogenic agents.

The effects of metabotropic glutamate receptor 7 allosteric agonist N,N'-dibenzhydrylethane-1,2-diamine dihydrochloride on developmental sevoflurane neurotoxicity: role of extracellular signal-regulated kinase 1 and 2 mitogen-activated protein kinase signaling pathway.

The present study was designed to evaluate the possible neuroprotective effects of metabotropic glutamate receptor (mGluR7) allosteric agonist N,N'-dibenzhydrylethane-1,2-diamine dihydrochloride (AMN082) on developmental sevoflurane neurotoxicity. To achieve the objective, hippocampal cultures (7 DIV, 7 day in vitro) were treated with different doses of L-(+)-2-amino-4-phosphonobutyric acid (L-AP4, an agonist of group III mGluRs), (RS)-α-Methylserine-O-phosphate (MSOP, an antagonist of group III mGluRs), AMN082 or cis-2-[[(3,5-dichlorophenyl)amino]carbonyl]cyclohexanecarboxylic acid (VU0155041, an agonist of mGluR4) before exposed to sevoflurane. Cell apoptosis were determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL)-staining. For in vivo study, rat pups (7 PND, 7 postnatal day) were injected with AMN082, L-AP4 or saline before sevoflurane exposure. Extracellular signal-regulated kinase 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38, caspase-3, Bcl-2, and Bax were detected by Western blot. The locomotor activity and cognitive functions were evaluated by open-field test and Morris water maze (MWM), respectively. We found that L-AP4 prevented sevoflurane-induced cell apoptosis, but MSOP promoted. Specially, application of AMN082 contributed to the relief of sevoflurane-induced apoptosis in vitro, whereas VU0155041 did not. In addition, sevoflurane treatment led to a decrease of Bcl-2 and an increase of caspase-3 and Bax, which were mitigated by AMNO82 in vivo. Moreover, we showed that sevoflurane treatment resulted in a remarkable suppression of phospho-ERK1/2, which was restored by AMN082. Application of U0126 (an inhibitor of MEK) abolished the neuroprotective effects of AMN082 on sevoflurane neurotoxicity both in vitro and in vivo. In addition, sevoflurane exposure also led to an increase of phospho-JNK, but SP600125 (an inhibitor of JNK) did not attenuate sevoflurane-induced apoptosis. The total and phosphorylated p38 remained unchanged in sevoflurane-treated rat pups. Finally, AMN082 improved the learning and memory defects caused by postnatal sevoflurane exposure without alternations in emotion or locomotor activity. These preliminary data indicate that AMN082 may protect immature brain against sevoflurane neurotoxicity, and the ERK1/2 MAP kinase signaling is likely to be involved. Further studies are needed to fully assess the neuroprotective role of mGluR7 agonist AMN082 in developmental anesthetic neurotoxicity.

Polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxinlike biphenyls in sediments from the Suzhou Creek, China.

Polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs) were detected in sediments from Suzhou Creek with mean concentrations of 478.1, 245.1, and 4727.6 pg/g dw, respectively. WHO-TEQ concentrations of PCDD/Fs in sediments ranged from 2.90 to 13.96 pg/g dw, while TEQ concentrations of PCBs varied from 0.27 to 1.41 pg/g dw. OCDD or HpCDD were the dominant congeners but PeCDF or HpCDD was the major contributor to PCDD/Fs-TEQ in all the sites. For dioxinlike biphenyls, PCB 118 was the major congener while PCB-TEQ was attributable to PCB 126 in all the samples.