ALPL regulates the aggressive potential of high grade serous ovarian cancer cells via a non-canonical WNT pathway.

The ALPL gene is linked to hypophosphatasia, a rare genetic disease. Owing to the inverse relationships between ALPL expression and both the International Federation of Gynecology and Obstetrics (FIGO) stages and histological grades assigned to patients with serous ovarian cancer (SOC), this study was designed to explore the role and possible mechanisms of ALPL in cell motility of high grade SOC (HGSOC). The effects of ALPL overexpression on migration and invasion were detected in HGSOC cell lines SKOV3 and HEY. Gene ontology analysis for differential genes with ALPL overexpression identified several biological processes, including EMT, correlated with cell motility. Genes potentially implicated in EMT and associated with ALPL were screened using The Cancer Genome Atlas (TCGA) database. The WNT receptor Frizzled2 (FZD2) was identified and its role in HGSOC cell motility and survival was investigated. It was found that forced expression of ALPL could inhibit migration, invasion, and EMT in HGSOC cells. It also reduced the expression of FZD2 and its ligand WNT5A, accompanied by suppressed expression of their downstream target phosphorylated-STAT3 (pSTAT3). These effects were initiated via the FZD2 knockdown using siRNA and reversed by recombinant WNT5A protein. The relationship between FZD2 expression and poor HGSOC patient survival was also investigated. This data supports that ALPL might restrict the function of WNT5A-FZD2-STAT3 axis, a non-canonical WNT pathway for promoting EMT progression, which results in attenuated migration and invasion in HGSOC cells and improves survival in patients with SOC.

[Value of serum gamma-glutamyl transpeptidase combined with direct bilirubin in the diagnosis of biliary atresia in infants].

OBJECTIVE: To study the value of serum gamma-glutamyl transpeptidase (GGT) combined with direct bilirubin (DB) in the diagnosis of biliary atresia. METHODS: A total of 667 infants with cholestasis who were hospitalized and treated from July 2010 to December 2018 were enrolled as subjects. According to the results of intraoperative cholangiography and follow-up, they were divided into biliary atresia group with 234 infants and cholestasis group with 433 infants. The two groups were compared in terms of age of onset, sex, and serum levels of total bilirubin (TB), DB, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acid (TBA), and GGT. A receiver operating characteristic (ROC) curve analysis was performed for indices with statistical significance, and the area under the ROC curve (AUC) and the optimal cut-off value for diagnosis were calculated. RESULTS: The biliary atresia group had a significantly younger age of onset than the cholestasis group (P<0.001). There were no significant differences in sex, ALT, and AST between the two groups (P>0.05), while the biliary atresia group had significantly higher serum levels of TB, DB, TBA, and GGT than the cholestasis group (P<0.05). GGT combined with DB had the highest AUC of 0.892 (95% confidence interval: 0.868-0.916) in the diagnosis of biliary atresia. At the optimal cut-off values of 324.0 U/L for GGT and 115.1 µmmol/L for DB, GGT combined with DB had a sensitivity of 79.8% and a specificity of 83.2% in the diagnosis of biliary atresia. CONCLUSIONS: GGT combined with DB has high sensitivity and specificity in the diagnosis of biliary atresia and can be used as an effective indicator for diagnosis of biliary atresia in infants.

Development of a novel quantitative function between spectral value and metmyoglobin content in Tan mutton.

This study was aimed to establish a quantitative function between spectral reflectance values and metmyoglobin (MetMb) content in Tan mutton during refrigeration. Near-infrared hyperspectral data combined with generalized two-dimensional correlation spectroscopy (G2D-COS) method to identify characteristic bands and investigate the sequence of chemical waveband changes. Characteristic wavebands identified by G2D-COS analysis had the best performance in predicting the content of MetMb, with a high R2p of 0.849, a low RMSEP of 2.695 and a high RPD of 2.786. The results showed that the G2D-COS may be a powerful tool for describing intensity changes of MetMb band. The partial least square regression method was used to develop the relationships between the spectral values and MetMb content in Tan mutton meat for predicting MetMb content. This study has provided a convenient and rapid non-destructive quantitative method for assessing the color of Tan mutton meat.

[Analysis of spectral response of vegetation leaf biochemical components].

Based on the leaf spectra data measured with LOPEX'93, the authors analyzed the relationship between leaf spectra and leaf biochemical components including leaf chlorophyll and leaf water content. It was showed that leaf chlorophyll was highly correlated with the second derivation of leaf reflectance at wavelengths 700, 670, 600, 500, 490, 440, and 410 nm; similarly, it was also showed that leaf water content was highly correlated with the continuum removed leaf reflectance at wavelengths 2350, 2180, 2130, 2120, 1870, and 1820 nm, which correspond to the absorption features of water in shortwave infrared bands. In addition, the authors found that ND (Normalized Difference) has a higher response to chlorophyll than other spectral indexes and the correlation coefficient is 0.618; Ratio975 has a very high response to water content and the correlation coefficient is up to 0.996. Based on the above evidences, the authors built a model to retrieve leaf chlorophyll and leaf water content through ground measured leaf spectra data and the simulated leaf chlorophyll and leaf water content are very accurate as compared to the ground measurements.

Glutamine-fructose-6-phosphate transaminase 2 (GFPT2) promotes the EMT of serous ovarian cancer by activating the hexosamine biosynthetic pathway to increase the nuclear location of ß-catenin.

The hexosamine biosynthetic pathway (HBP), a branch of glucose metabolism, provides a substrate for glycosylation modification, which has a wide-ranging effect on cellular functions. Glutamine-fructose-6-phosphate transaminase 2 (GFPT2) has been reported to regulate the HBP as the first and rate-limiting enzyme. Given the inverse association between GFPT2 expression and survival of patients with serous ovarian cancer (SOC) observed in The Cancer Genome Atlas (TCGA) database, we attempted to investigate the role of GFPT2 and its related mechanisms in SOC. The results showed that GFPT2 was over-expressed in SOC tissues, and positive correlations with advanced stage (FIGO III/IV), suboptimal removal rate and poor survival were observed in 90 SOC patients. Cell migration and invasion were also inhibited in GFPT2 knockdown SKOV3 and HEY cells. The levels of O-linked ß-N-acetylglucosamine (O-GlcNAc) and intranuclear ß-catenin were evaluated and the observed increase in O-GlcNAcylation induced by GFPT2 may contribute to epithelial-mesenchymal transition (EMT). These data provide novel insights into the function of GFPT2 and O-GlcNAcylation in the EMT and thus the invasiveness SOC.

[Phytoremediation of Petroleum Contaminated Soils with Iris pseudacorus L. and the Metabolic Analysis in Roots].

In this study, we performed a greenhouse pot-culture experiment to investigate the potential of a wild ornamental plant Iris pseudacorus L. in remediating petroleum contaminated soils from the Dagang Oilfield in Tianjin, China. The results suggested that Iris pseudacorus L. had great resistance to ≤ 40,000 mg · kg(⁻¹ of total petroleum hydrocarbons (TPHs). The removal rate of TPHs with concentrations of 10,000 mg · kg⁻¹, 20,000 mg · kg⁻¹ and 40,000 mg · kg⁻¹ in soils by Iris pseudacorus L. was 42.1%, 33.1% 31.2%, respectively, much higher than those in the corresponding controls (31.8%, 21.3% 11.9%, respectively) (P < 0.05). The root specific surface area of Iris pseudacorus L. was determined by the root scanner. The results suggested that TPHs with concentrations of 10,000 mg · kg⁻¹, 20,000 mg · kg⁻¹ and 40,000 mg · kg⁻¹ in soils increased the root specific surface area comparing with the controls. Additionally, the metabolic analysis showed that root metabolism changed to different degrees under the stress of TPHs, and the levels or species of metabolites had a significant change (P < 0.001). Furthermore, the results showed that 5 of 11 metabolites (VIP value > 1.2) with the root specific surface area from the PLS-DA model analysis, including ethanedioic acid, lactic acid, 2-butenedioic acid, phosphate and propanedioic acid, were positively correlated with the root specific surface area, but the others, gluconic acid, uridine, butanoic acid, maltose, 9,12-octadecadienoic acid, phenylalanine, were negatively correlated with it. In conclusion, using Iris pseudacorus L. to remediate petroleum contaminated soils is feasible, and the metabolic analysis in roots is useful to better understand the metabolic response of plants exposure to petroleum contaminated soils, and then reveals its remediated mechanisms.

[Rhizospheric Mechanisms of Hemerocallis middendorfii Trautv. et Mey. Remediating Petroleum-contaminated Soil and Metabonomic Analyses of the Root Systems].

The effects of a special ornamental plant Hemerocallis middendorfii Trautv. et Mey. on remediating petroleum-contaminated soil from the Dagang Oilfield in Tianjin, China, was studied by a greenhouse pot-culture experiment and the gradients of TPHs were 0, 10,000 and 40,000 mg · kg⁻¹. The results suggested that H. middendorfii had a high tolerance to TPHs (≤ 40,000 mg · kg⁻¹). And H. middendorfii significantly (P < 0.05) promoted the removal rate of TPHs (53.7% and 33.4%) compared with corresponding controls (31.8% and 12.0%) by natural degradation, respectively. The relative abundance of amino acids, organic acids and sugars and others in soil were analyzed by gas chromatography-mass spectrometry (GC-MS), and PCA and PLS-DA models were to investigate the rhizospheric mechanisms. The results suggested that H. middendorfii changed the distribution characteristics of each component in soil, and the glucopyranoside played a key role in the removal of TPHs. Furthermore, the results about comparative metabolic profile showed that some special metabolites were only found in the contaminated groups, including alanine, tetradecanoic acid, hexadecanoic acid and 9,12-octadecadienoic acid. Additionally, the exposure of TPHs changed the primary metabolic flux of roots, and caused the significant (P < 0.01) change of metabolites. In conclusion, H. middendorfii might be an enduring ornamental plant for effective remediating TPHs (≤ 40,000 mg · kg⁻¹) in soil. But the exposure of TPHs had changed the metabolic profile of H. middendorfii in roots, which might be the metabolic response of H. middendorfii to petroleum-contaminated soil.

Effects of comprehensive intensive therapies on the change of intima-media thickness of carotid arteries in type 2 diabetic patients: A report of 4-year follow-up with a literature review.

BACKGROUND: The aim was to evaluate the effect of comprehensive intensive therapy on the carotid and femoral arteries of intima-media thickness in patients with type 2 diabetes mellitus after 4-year follow-up. METHODS: In this prospective 4-year study, patients (N = 210) with newly diagnosed type 2 diabetes received either comprehensive intensive therapy (n = 110) or conventional therapy (n = 100). Blood pressure, blood glucose and lipid levels were monitored every 3-6 months, and carotid and femoral arteries of intima-media thickness were monitored with ultrasonography. For the literature review, various databases were searched until 20 December 2014 for studies that evaluated effects of intensive multi-factorial therapies on comprehensive intensive therapy in type 2 diabetes mellitus patients. RESULTS: The comprehensive intensive therapy group had a smaller rate of carotid intima-media thickness increase than the conventional therapy (control) group (p < 0.05). The carotid intima-media thickness in comprehensive intensive therapy group remained stable while the adjusted rate of carotid intima-media thickness increase was 12.55% in the control group. The femoral intima-media thickness change was also smaller in comprehensive intensive therapy group but the difference over time did not reach significance. CONCLUSION: The carotid intima-media thickness remained stable in type 2 diabetes mellitus patients who received comprehensive intensive therapy, suggesting that multi-factorial intensive therapies might have potential in reducing macro-vascular events in these patients.

[Stable suppression of beta-catenin expression in prostate cancer cell line by retrovirus mediated RNAi].

OBJECTIVE: To set up a prostate cancer cell line in which beta-catenin expression is stably suppressed and to investigate the role of Wnt/beta-catenin signaling pathway in prostate tumorgenesis. METHODS: We select 3 sites in the complete coden sequence region of beta-catenin gene as the RNAi targets, ligated the annealed double pre-DNA strands into the retroviral vectors pSUPER-retro and transfected them into the packaging cells PA317, and then collected supernatant with retrovirus to infect DU145. After selection by puromycin and culture expansion, the stable cell clones were attained. Expression of the 2 target genes of Wnt/beta-catenin signaling pathway cyclinD1 and c-myc, was detected in the beta-catenin RNAi cells by Western blot. The effect of suppressing beta-catenin by RNAi on cell proliferation was quantified by methylthiazoletetrazolium (MTT) assay. RESULTS: Western blotting and RT-PCR showed that the expression level of beta-catenin in the 2 stable cell clones apparently decreased. CyclinD1 and c-myc expression decreased in the beta-catenin RNAi cells. MTT showed that the cell number of beta-catenin expression suppression cell clones decreased significantly (P < 0. 05), suggesting the cell proliferation was prevented. CONCLUSION: The beta-catenin gene stable suppression cell line was successfully established.

An increase in the cerebral infarction area during fatigue is mediated by il-6 through an induction of fibrinogen synthesis.

OBJECTIVES: Our study aimed to investigate the impact of fatigue on the severity of stroke and to explore the underlying mechanisms. METHODS: Fatigued male rats underwent middle cerebral artery occlusion and the infarcted brain area was determined. Then, coagulation parameters were assessed in the fatigued group and a control group. In addition, the level of fibrinogen was determined in rats deprived of sleep for various numbers of days. To study whether interleukin-6 was involved in fibrinogen synthesis during fatigue, we also measured levels of interleukin-6 in rats deprived of sleep for various numbers of days. Furthermore, brain injury by middle cerebral artery occlusion was measured in wild-type mice, interleukin-6-/- mice and wild-type mice treated with bezafibrate. RESULTS: More severe cerebral infarction was observed in the fatigued rats, resulting in an infarct ratio of 23.4%. The infarct ratio was significantly increased in the fatigued rats compared with that in the control group (8%, p<0.05). The level of fibrinogen was increased significantly in the fatigued rats compared with that in the control group. In addition, a marked reduction in fibrinogen level was observed in the fatigued interleukin-6-/- mice compared to their wild-type counterparts, whereas no difference was observed between fatigued wild-type mice and interleukin-6-/- rats treated with recombinant human interleukin-6. The reduction in brain injury due to middle cerebral artery occlusion during fatigue was observed in interleukin-6-/- mice and wild-type mice treated with bezafibrate. CONCLUSION: Fatigue could increase stroke severity and was associated with the interleukin-6-induced expression of fibrinogen.

An increase in the cerebral infarction area during fatigue is mediated by il-6 through an induction of fibrinogen synthesis

OBJECTIVES: Our study aimed to investigate the impact of fatigue on the severity of stroke and to explore the underlying mechanisms. METHODS: Fatigued male rats underwent middle cerebral artery occlusion and the infarcted brain area was determined. Then, coagulation parameters were assessed in the fatigued group and a control group. In addition, the level of fibrinogen was determined in rats deprived of sleep for various numbers of days. To study whether interleukin-6 was involved in fibrinogen synthesis during fatigue, we also measured levels of interleukin-6 in rats deprived of sleep for various numbers of days. Furthermore, brain injury by middle cerebral artery occlusion was measured in wild-type mice, interleukin-6-/- mice and wild-type mice treated with bezafibrate. RESULTS: More severe cerebral infarction was observed in the fatigued rats, resulting in an infarct ratio of 23.4%. The infarct ratio was significantly increased in the fatigued rats compared with that in the control group (8%, p<0.05). The level of fibrinogen was increased significantly in the fatigued rats compared with that in the control group. In addition, a marked reduction in fibrinogen level was observed in the fatigued interleukin-6-/- mice compared to their wild-type counterparts, whereas no difference was observed between fatigued wild-type mice and interleukin-6-/- rats treated with recombinant human interleukin-6. The reduction in brain injury due to middle cerebral artery occlusion during fatigue was observed in interleukin-6-/- mice and wild-type mice treated with bezafibrate. CONCLUSION: Fatigue could increase stroke severity and was associated with the interleukin-6-induced expression of fibrinogen. .