LncRNA-NEAT1 blocks the Wnt/ß-catenin signaling pathway by targeting miR-217 to inhibit trophoblast cell migration and invasion.

OBJECTIVE: This study aimed to study the correlation between preeclampsia (PE) and lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1), and to examine the molecular mechanisms behind the development of PE. METHODS: 30 PE and 30 normal pregnant women placental samples were assessed the levels of NEAT1 and miR-217 by quantitative real-time PCR (qRT-PCR). The trophoblast cell line HTR8/SVneo was used for silencing NEAT1 or miR-217 inhibitor in the absence or presence of an inhibitor and H2O2. Cell counting Kit 8 (CCK-8), flow cytometry, and Transwell were used to detect cell proliferation, apoptosis, migration, and invasion. Luciferase reporter gene assay was utilized to verify the binding between miR-217 and Wnt family member 3 (Wnt3), and between the miR-217 and NEAT1. Proteins related to the Wnt/ß-catenin signaling pathway were detected using western blotting. RESULTS: The PE group exhibited a significantly downregulated expression of miR-217 and a significantly upregulated expression of NEAT1. NEAT1 targeted miR-217, and Wnt is a miR-217 target gene. siRNA-NEAT1 inhibited the apoptosis of trophoblast cells, but promoted their invasion, migration, and proliferation. MiR-217 inhibitor could partially reverse the effects of siRNA-NEAT1. The expression of the Wnt/ß-catenin signaling pathway-related proteins, WNT signaling pathway inhibitor 1 (DKK1), cyclin-D1 and ß-catenin, was significantly increased after siRNA-NEAT1. CONCLUSIONS: NEAT1 could reduce trophoblast cell invasion and migration by suppressing miR-217/Wnt signaling pathway, leading to PE.

2.79-µm efficient acousto-optic Q switched Er,Cr:YSGG laser with an LiNbO3 crystal modulator.

We fabricate a novel, to the best of our knowledge, acousto-optic Q switch in the 3-micron wavelength range using a longitudinal acoustic mode in a lithium niobate (LiNbO3) crystal. The device is designed based on properties of the crystallographic structure and material to obtain a high diffraction efficiency close to the theoretical prediction. The effectiveness of the device is verified by application in an Er,Cr:YSGG laser at 2.79 µm. The maximum diffraction efficiency reached 57% at the radio frequency of 40.68 MHz. At the repetition rate of 50 Hz, the maximum pulse energy was 17.6 mJ and the corresponding pulse width was 55.2 ns. The effectiveness of bulk LiNbO3 as an acousto-optic Q switch is verified for the first time.

Toxicity Assessment of 4 Azo Dyes in Zebrafish Embryos.

Azo dyes are used widely as color additives in food, drugs, and cosmetics; hence, there is an increasing concern about their safety and possible health hazards. In the present study, we chose 4 azo dyes tartrazine, Sunset Yellow, amaranth, and Allura red and evaluated their developmental toxicity on zebrafish embryos. At concentration levels of 5 to 50 mM, we found that azo dyes can induce hatching difficulty and developmental abnormalities such as cardiac edema, decreased heart rate, yolk sac edema, and spinal defects including spinal curvature and tail distortion. Exposure to 100 mM of each azo dye was completely embryolethal. The median lethal concentration (LC50), median effective concentration (EC50), and teratogenic index (TI) were calculated for each azo dye at 72 hours postfertilization. For tartrazine, the LC50 was 47.10 mM and EC50 value was at 42.66 mM with TI ratio of 1.10. For Sunset Yellow, the LC50 was 38.93 mM and EC50 value was at 29.81 mM with TI ratio of 1.31. For amaranth, the LC50 was 39.86 mM and EC50 value was at 31.94 mM with TI ratio of 1.25. For Allura red, the LC50 was 47.42 mM and EC50 value was 40.05 mM with TI ratio of 1.18. This study reports the developmental toxicity of azo dyes in zebrafish embryos at concentrations higher than the expected human exposures from consuming food and drugs containing azo dyes.

Developmental toxicity of auranofin in zebrafish embryos.

Auranofin (AF) is used in clinic for the treatment of rheumatoid arthritis, repurposing of AF as an anticancer drug has just finished a phase I/II clinical trial, but the developmental toxicity of AF remains obscure. This study focused on its developmental toxicity by using zebrafish embryos. Zebrafish embryos were exposed to different concentrations (1, 2.5, 5, 10 µm) of AF from 2 h post-fertilization (hpf) to 72 hpf. At 72 hpf, two major developmental defects caused by AF were found, namely severe pericardial edema and hypopigmentation, when embryos were exposed to concentrations higher than 2.5 µm. Biochemical detection of oxidative stress enzyme combined with expressions of a series of genes related to oxidative stress, cardiac, metal stress and pigment formation were subsequently tested. The superoxide dismutase activity was decreased while malondialdehyde content was accumulated by AF treatment. The expression of oxidative stress-related genes (sod1, gpx1a, gst), pigment-related genes (mitfb, trp-1a) and one metal stress-related gene ctr1 were all decreased by AF exposure. The expressions of cardiac-related genes (amhc, vmhc) and one metal-related gene hsp70 were found to be significantly upregulated by AF exposure. These findings indicated the potential developmental toxicity of AF on zebrafish early development. Copyright © 2016 John Wiley & Sons, Ltd.

[Separation of Chlorophyll Fluorescence from Scattering Light of Algal Water Based on the Polarization Technique].

Once chlorophyll molecules within the planktonic algae cells absorb light energy, they will release energy in the form of chlorophyll fluorescence emission. The elastic scattering light of particles in water is partially polarized, while chlorophyll fluorescence is unpolarized, so we can use the properties to separate the fluorescence signal from total scattering spectra for further retrieval the concentration of chlorophyll. But for coastal waters, the approach feasibility is still unclear. Based on this, we conducted the experiment in laboratory to analyze the influence of different concentrations of inorganic particles (IOP) and chlorophyll on the fluorescence extraction using polarization discrimination technique. The results indicate that, for algae water with different concentrations of IOP, the fluorescence peak will decrease while the concentration increase, but the retrieval result is still reliable when the concentration up to 300 mg·L(-1). For algae water with different concentrations of chlorophyll, the concentration of chlorophyll more higher, the efficiency of extracted fluorescence using polarization method is better, for common water, this approach can still work. The study proves that the polarization method is also applicable for complex water; it is of great importance for further detecting the concentration of chlorophyll in coastal waters using remote sensing.

Mycophenolic Acid-Induced Developmental Defects in Zebrafish Embryos.

With the increasing use of mycophenolic acid (MPA) in solid organ transplantation, some clinical studies indicate that it is also a human teratogen. However, it is unknown by which mechanism MPA acts as a teratogen. Mycophenolic acid was a selective blocker of de novo purine synthesis, and its immunosuppressive effect is mediated by the inhibition of inosine monophosphate dehydrogenase, which could be a target for MPA-induced toxicity as well. The aim of our study was to examine the direct influence of MPA exposure on zebrafish (Danio rerio) embryos. Morphological defects including tail curvature and severe pericardial edema in zebrafish embryos caused by MPA (3.7-11.1 µmol/L) were found in a dose-dependent manner. The teratogenic index (25% lethal concentration value (LC25)/no observed adverse effect level ratio) was 16, which indicated MPA as a teratogen. Quantitative polymerase chain reaction analysis revealed that the expression level of impdh1b and impdh2 was significantly reduced by MPA treatment at 8 µmol/L (equals to LC25 level). All the toxic effects could be partially reversed by the addition of 33.3 µmol/L guanosine. Our results indicated that MPA impairs the development of zebrafish embryos via inhibition of impdh activity, which subsequently caused a guanosine nucleotide depletion in vivo.

Expression of advanced glycation end products in placenta and concentration in maternal and umbilical serum in pre-eclampsia.

AIM: To study the function of free fatty acid (FFA) in maternal serum and advanced glycation end products (AGE) in maternal serum, umbilical cord blood and in the placenta of women with pre-eclampsia. METHODS: Sixty women with pre-eclampsia and 60 normal pregnant women as controls participated in this study. Women with pre-eclampsia were divided into an early-onset group and late-onset group, with 30 normal pregnant women as the early control group and 30 as the late control group. Competitive enzyme-linked immunosorbent assay was used to measure AGE in maternal serum and umbilical cord blood. Western blotting was performed to analyze AGE in the placenta. Improved copper agent colorimetry was performed to measure FFA in maternal serum. RESULTS: Mean AGE in maternal serum in the early-onset and late-onset pre-eclampsia groups was significantly higher than in the control groups. Mean concentration of umbilical cord blood AGE was significantly higher in the early-onset and late-onset pre-eclampsia groups when compared with the control groups. On western blot the level of placental AGE in the early-onset and late-onset pre-eclampsia groups was significantly higher than in normal placenta. Serum FFA in the early-onset and late-onset pre-eclampsia groups was higher than in healthy pregnant women. Maternal serum FFA in the early-onset and late-onset pre-eclampsia groups correlated positively with maternal serum AGE (r = 0.764 and r = 0.774, P < 0.05). CONCLUSIONS: AGE and FFA, which are upregulated in pre-eclampsia, are likely to be involved in pre-eclampsia through some common mechanism.

[Backscattering characteristics of Amphidinium carterae Hulburt].

Variability of the backscattering characteristics of the alga amphidinium carterae hulburt and its interpretation were examined using the measured data performed during July 2012 in the laboratory. The results indicated that the backscattering coefficient has the characteristic of spectral variability, and was highly power correlated with the chlorophyll concentration, the minimum of the correlation coefficient reached up to 0.96; otherwise, the shape of the particulate backscattering coefficient spectra was also changed from the low chlorophyll concentration to the higher concentration because of the influence of chlorophyll absorption. At the same time, the authors got another value of the particulate backscattering ratio ranging between 0.006 4 and 0.011 6 at 620 nm, although a global increase in particulate backscattering ratio with growing chlorophyll concentration was also visible, but the regularity is not obviously especially at the higher chlorophyll concentration and unlike backscattering coefficient, the spectral shapes of the backscattering ratio remained constant. The authors also found that particle density was one of the main factors controlling the variability of particles backscattering ratio for the pure algal water, and the authors fit a power law function to them and the correlation coefficient at 620 nm was as high as 0.98.

High expression of peroxisomal D-bifunctional protein in cytosol regulates apoptosis and energy metabolism of hepatocellular carcinoma cells via PI3K/AKT pathway.

Peroxisomal D-bifunctional protein (DBP) is an indispensable enzyme of the fatty acid ß-oxidation in the peroxisome of humans. However, the role of DBP in oncogenesis is poorly understood. Our previous studies have demonstrated that DBP overexpression promotes hepatocellular carcinoma (HCC) cell proliferation. In this study, we evaluated the expression of DBP in 75 primary HCC samples using RT-qPCR, immunohistochemistry, and Western blot, as well as its correlation with the prognosis of HCC. In addition, we explored the mechanisms by which DBP promotes HCC cell proliferation. We found that DBP expression was upregulated in HCC tumor tissues, and higher DBP expression was positively correlated with tumor size and TNM stage. Multinomial ordinal logistic regression analysis indicated that lower DBP mRNA level was an independent protective factor of HCC. Notably, DBP was overexpressed in the peroxisome and cytosol and mitochondria of tumor tissue cells. Xenograft tumor growth was promoted by overexpressing DBP outside peroxisome in vivo. Mechanistically, DBP overexpression in cytosol activated the PI3K/AKT signaling axis and promoted HCC cell proliferation by downregulating apoptosis via AKT/FOXO3a/Bim axis. In addition, overexpression of DBP increased glucose uptake and glycogen content via AKT/GSK3ß axis, as well as elevated the activity of mitochondrial respiratory chain complex III to increase ATP content via the mitochondrial translocation of p-GSK3ß in an AKT-dependent manner. Taken together, this study was the first to report the expression of DBP in peroxisome and cytosol, and that the cytosolic DBP has a critical role in the metabolic reprogramming and adaptation of HCC cells, which provides a valuable reference for instituting an HCC treatment plan.

[Double balloon endoscopy in diagnosis of patients with obscure gastrointestinal bleeding].

OBJECTIVE: To assess the diagnostic value of double balloon endoscopy (DBE) for obscure gastrointestinal bleeding (OGIB) METHODS: The data of 103 OGIB patients who underwent DBE from January 2007 to September 2010 in the First Affiliated Hospital, Zhejiang University School of Medicine were retrospectively analyzed. RESULTS: DBE was successfully performed in all 103 patients without complications. Of 103 patients, 66(64.1 %) had positive DBE findings and 28 had surgery procedures(27.2 %). Ninety-four patients finally acquired positive diagnosis, including small intestine tumor(31.1 %), angiodysplasia(22.3 %), exulceratio simplex(9.7 %), Crohn's disease(6.8 %), diverticulum(4.9 %), abdominal purpure(4.9 %), etc. Lesions occurred more frequently in proximal small intestine than in distal small intestine (56.3 % Compared with 30.1 %, P<0.001). CONCLUSION: DBE is a safe, effective and reliable procedure for the diagnosis of OGIB.

Associations between polymorphisms in VDR gene and the risk of osteoporosis: a meta-analysis.

The authors conducted this meta-analysis to robustly estimate relationships between polymorphisms in VDR gene and the risk of osteoporosis by integrating the results of previous works. Medline, Embase, Wanfang, VIP and CNKI were searched thoroughly for eligible studies, and 73 genetic association studies were finally included in this meta-analysis. We noticed that ApaI rs7975232, BsmI rs1544410 and TaqI rs731236 polymorphisms were significantly associated with the risk of osteoporosis in Caucasians. Moreover, BsmI rs1544410 and FokI rs10735810 polymorphisms were significantly associated with the risk of osteoporosis in Asians. In conclusion, this meta-analysis demonstrates that ApaI rs7975232, BsmI rs1544410 and TaqI rs731236 polymorphisms may influence the risk of osteoporosis in Caucasians, while BsmI rs1544410 and TaqI rs731236 polymorphisms may influence the risk of osteoporosis in Asians.

Inhibitory effect of main phenolic acid components of Jacobaea cannabifolia (Less.) on inflammation caused by PM2.5.

PM2.5 is an important environmental problem threatening human health at present, which poses serious harm to human body after inhalation. J. cannabifolia is a traditional Chinese medicine which exhibits anti-inflammatory effect. This study aimed to investigate the inhibitory effect of main phenolic acid components of J. cannabifolia on inflammation caused by PM2.5. Effect of PM2.5 on cell activity and apoptosis were determined by MTT, flow cytometry and calcein AM/PI staining. PHBA, PHPAA, and mixture of PHBA and PHPAA of different concentrations were given to RAW264.7 cells pretreated with PM2.5. The effect of drugs on cellular inflammatory factors was detected by ELISA. The expressions of TLRs related signal pathway at protein and gene levels were detected by western blot and qRT-PCR. The results showed that PM2.5 had no effect on cell activity and apoptosis within the determined concentration range. PHBA and PHPAA could markly inhibit the level of IL-1ß, IL-6, and TNF-α in RAW264.7 cells. Furthermore, the expressions of TLR2, TLR4, MyD88, IRAK1, TRAF6, TAK1, IKKß, and NF-κB induced by PM2.5 were markedly inhibited by PHBA and PHPAA at protein and gene levels. This study demonstrated that PHBA and PHPAA could attenuated inflammation caused by PM2.5 through suppressing TLRs related signal pathway.

Association between the reactivity of local cerebral oxygen saturation after hypo-to-hypercapnic tests and delirium after abdominal surgery in older adults: A prospective study.

Objective: This study aimed to investigate the correlation between changes in regional cerebral oxygen saturation (rSO2) and postoperative delirium in older adults undergoing major abdominal surgery. Materials and methods: This prospective study enrolled older adults scheduled for elective major abdominal surgery at the Second Affiliated Hospital of Anhui Medical University from August 2021 to January 2022. The change in rSO2 from baseline was determined using the hypo-to-hypercapnic test. The main study outcome was the occurrence of postoperative delirium. Results: A total of 101 participants were included for analysis, of whom 16 (15.8%) developed postoperative delirium. Compared with non-delirium participants, the mean arterial pressure and heart rate were not significantly different in the postoperative delirium group at T0, T1, T2, T3, T4, and T6 (all Pinteraction > 0.05), but the delirium group had lower pH, lower PaO2, and higher lactate levels at T4, T5, and T6 (all Pinteraction < 0.05). rSO2 at T0, T1, T2, T3, T4, and T6 was 69.0 (63.2-75.2), 70.7 ± 7.3, 68.2 ± 7.5, 72.1 ± 8.0, 69.9 ± 7.8, 67.4 ± 7.2, and 71.7 ± 8.1, respectively. The postoperative change in rSO2 during the hypercapnia test (TΔrSO2%) was 6.62 (5.31-9.36). Multivariable analysis showed that the Cumulative Illness Rating Scale (odd ratio, OR = 1.89, 95% confidence interval, CI: 1.10-3.25, P = 0.021), preoperative albumin levels (OR = 0.67, 95% CI: 0.48-0.94, P = 0.022), rSO2 at T4 (OR = 0.61, 95% CI: 0.41-0.89, P = 0.010), and postoperative TΔrSO2% (OR = 0.80, 95% CI: 0.66-0.98, P = 0.028) were independently associated with postoperative delirium in older adults undergoing elective abdominal surgery. Conclusion: The rSO2 measured at T4 and postoperative TΔrSO2% were independently associated with postoperative delirium in older adults undergoing elective abdominal surgery.

Prediction of pre-eclampsia complicated by fetal growth restriction and its perinatal outcome based on an artificial neural network model.

Objective: Pre-eclampsia (PE) complicated by fetal growth restriction (FGR) increases both perinatal mortality and the incidence of preterm birth and neonatal asphyxia. Because ultrasound measurements are bone markers, soft tissues, such as fetal fat and muscle, are ignored, and the selection of section surface and the influence of fetal position can lead to estimation errors. The early detection of FGR is not easy, resulting in a relative delay in intervention. It is assumed that FGR complicated with PE can be predicted by laboratory and clinical indicators. The present study adopts an artificial neural network (ANN) to assess the effect and predictive value of changes in maternal peripheral blood parameters and clinical indicators on the perinatal outcomes in patients with PE complicated by FGR. Methods: This study used a retrospective case-control approach. The correlation between maternal peripheral blood parameters and perinatal outcomes in pregnant patients with PE complicated by FGR was retrospectively analyzed, and an ANN was constructed to assess the value of the changes in maternal blood parameters in predicting the occurrence of PE complicated by FGR and adverse perinatal outcomes. Results: A total of 15 factors-maternal age, pre-pregnancy body mass index, inflammatory markers (neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio), coagulation parameters (prothrombin time and thrombin time), lipid parameters (high-density lipoprotein, low-density lipoprotein, and triglyceride counts), platelet parameters (mean platelet volume and plateletcrit), uric acid, lactate dehydrogenase, and total bile acids-were correlated with PE complicated by FGR. A total of six ANNs were constructed with the adoption of these parameters. The accuracy, sensitivity, and specificity of predicting the occurrence of the following diseases and adverse outcomes were respectively as follows: 84.3%, 97.7%, and 78% for PE complicated by FGR; 76.3%, 97.3%, and 68% for provider-initiated preterm births,; 81.9%, 97.2%, and 51% for predicting the severity of FGR; 80.3%, 92.9%, and 79% for premature rupture of membranes; 80.1%, 92.3%, and 79% for postpartum hemorrhage; and 77.6%, 92.3%, and 76% for fetal distress. Conclusion: An ANN model based on maternal peripheral blood parameters has a good predictive value for the occurrence of PE complicated by FGR and its adverse perinatal outcomes, such as the severity of FGR and preterm births in these patients.

Differences in Gall Induction of Flower-like Galls on Haloxylon by Psyllids (Hemiptera: Aphalaridae), and the Emergence of Corresponding Parasitoids.

Saxaul is a kind of dominant perennial psammophyte that widely distributes in arid and semi-arid desert areas, and it has multiple functions in preventing desertification, especially in windbreak and sand fixation. Various gall inducers induce galls on the saxaul, including the flower-like gall. Parasitoids have great potentiality in controlling gall inducers. However, studies about gall inducers and parasitoids of flower-like galls on Haloxylon, as well as the parasitic efficacy of the parasitoids, are rarely reported. In this study, the flower-like galls were observed on Haloxylon ammodendron and H. persicum in Fukang, Xinjiang, China. Two types of flower-like galls were found on H. ammodendron, while only one type was found on H. persicum. In total, five species of gall inducers and three species of parasitoids were obtained from the galls mentioned above. All the galls were induced by Caillardia (Hemiptera: Aphalaridae), which were mostly bivoltine in Fukang. Besides, their parasitoids Psyllaephaguscaillardiae and P. longiventris could be observed on all the types of galls. Additionally, correlative studies on the parasitization indexes demonstrated that all the dominant parasitoids of diverse flower-like galls were P. caillardiae, which were slightly more in number than the P. ogazae discovered in the flower bud-like galls. In addition, the relevance between the emergence or lifespan of parasitoids and temperature was also investigated. The results showed that the number of parasitoids emerging decreased rapidly after a period of enhancement with the increase of temperature, including an optimum temperature, while the lifespan of wasps gradually shortened with the temperature rising. Our results highlight the importance of the biological investigation of parasitoids in the gall inducers lived in closed galls, which may provide critical evidence for us to understand its potential application in biological control.

Mechanism of hepatotoxicity of first-line tyrosine kinase inhibitors: Gefitinib and afatinib.

AIMS: Both gefitinib and afatinib are epidermal growth factor tyrosine kinase inhibitors (EGFR-TKI) in the treatment of non-small cell lung cancer (NSCLC). It has been reported that gefitinib and afatinib could cause hepatotoxicity during the clinic treatment, therefore it is critical to investigate their hepatotoxicity systematically. In this study, zebrafish (Danio rerio) were used as model animals to compare the hepatotoxicity and their toxic mechanism. MAIN METHODS: The zebrafish transgenic line [Tg (fabp10a: dsRed; ela3l:EGFP) was used in this study. After larvae developed at 3 days post fertilization (dpf), they were put into different concentrations of gefitinib and afatinib. At 6 dpf, the viability, liver area, fluorescence intensity, histopathology, apoptosis, transaminase reflecting liver function, the absorption of yolk sac, and the expression of relative genes were observed and analyzed respectively. KEY FINDINGS: Both gefitinib and afatinib could induce the larvae hepatotoxicity dose-dependently. Based on the liver morphology, histopathology, apoptosis and function assessments, gefitinib showed higher toxicity, causing more serious liver damage. Both gefitinib and afatinib caused abnormal expressions of genes related to endoplasmic reticulum stress (ERS) pathway and apoptosis. For example, jnk, perk, bip, chop, ire1, bid, caspase3 and caspase9 were up-regulated, while xbp1s, grp78, bcl-2/bax, and caspase8 were down-regulated. The hepatotoxicity difference of gefitinib and afatinib might be due to the different expression level of related genes.

Diagnosis of obscure gastrointestinal hemorrhage with capsule endoscopy in combination with multiple-detector computed tomography.

AIM: To demonstrate the clinical efficacy of combination capsule endoscopy (CE) and multiple-detector computed tomography (MDCT) diagnostic imaging in the identification of gastrointestinal hemorrhages. METHODS: In the present study, 123 patients with gastrointestinal hemorrhages of obscure origin (GHOO) were examined with CE in combination with MDCT. The results were compared with findings of surgical pathology. RESULTS: Of the 123 patients, 57.72% (71/123) of the patients exhibited positive CE findings compared with 30.08% (37/123) on MDCT alone (P < 0.01). When used in combination, 65.85% (81/123) of patients scored positively. The detection rate due to the combination of diagnostic imaging was significantly higher than that of MDCT alone (P < 0.01), but was not significantly higher than that of CE alone (P > 0.05). Integrating the two diagnostic platforms improved the diagnosis of stromal tumors, hemangioma, Crohn's disease, vascular anomaly, Meckel's diverticulum, and ancylostomiasis. There was no significant difference in the positive detection rate between CE and MDCT when confirmed by surgical pathology. CONCLUSION: The contribution of CE is critical in the diagnosis of GHOO, given the fact that there is a significant difference in the detection rate between CE and MDCT, but there is no significant difference in the rate between CE plus MDCT and CE alone.

De-regulated STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis suppresses CML cell apoptosis and contributes to Imatinib resistance.

BACKGROUND: STAT5 plays an important role in the transformation of hematopoietic cells by BCR-ABL. However, the downstream target genes activated by STAT5 in chronic myeloid leukemia (CML) cells remain largely unclear. Here, we investigated the mechanistic functional relationship between STAT5A-regulated microRNA and CML cell apoptosis. METHODS: The expression of USP15, Caspase-6, STAT5A-regulated miR-202-5p and STAT5A was detected by qRT-PCR and Western blotting in CML cell lines and PBMCs of CML patients. Cell apoptosis was evaluated by flow cytometry. Both gain- and loss-of-function experiments were used to investigate the roles of USP15, miR-202-5p and STAT5A in CML. Luciferase reporter assay detected the effect of miR-202-5p on USP15 expression. Xenograft animal model was used to test the effect of anti-miR-202-5p and pimozide on K562 cell xenograft growth. RESULTS: USP15 expression was significantly downregulated in CML cell lines and PBMCs of CML patients. Depletion of USP15 increased, whereas overexpression of USP15 reduced the resistance of CML cells to Imatinib. Further, decreased deubiquitinating activity of USP15 by USP15 downregulation led to reduced caspase-6 level, thus attenuating CML cell apoptosis. Mechanistically, miR-202-5p was upregulated in K562G cells and negatively regulated USP15 expression by directly targeting USP15 3'-UTR. Correspondingly, upregulation of miR-202-5p enhanced the resistance of CML cells to Imatinib by inhibiting cell apoptosis. Importantly, STAT5A was upregulated in CML cells and directly activated miR-202-5p transcription by binding to the pre-miR-202 promoter. Pimozide induced CML cell apoptosis and significantly reduced K562 cell xenograft growth in vivo by blocking STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis. CONCLUSIONS: we provide the first evidence that de-regulated STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis suppresses the apoptosis of CML cells, targeting this pathway might be a promising therapeutic approach for the treatment of CML.

Local mild hypothermia induced by intra-arterial cold saline infusion prolongs the time window of onset of reperfusion injury after transient focal ischemia in rats.

OBJECTIVES: The aims of this study were to determine the effects of intra-arterial local hypothermia on infarct volume in rats with different durations of ischemia and to determine whether hypothermia can prolong the therapeutic time window compared with reperfusion without hypothermia. METHODS: Adult male Sprague-Dawley rats weighing 260-300 g were divided into control group (permanent MCA occlusion), normothermia groups (NT groups) and hypothermia groups (HT groups). NT groups included rats induced with blood reperfusion for 1.5, 2, 2.5 or 3 hour ischemia. In the HT groups with ischemia of 1.5, 2, 2.5 or 3 hours, 6 ml 20 degrees C normal saline solution was flushed at a speed of 0.6 ml/min, beginning 10 minutes before blood reperfusion. The infarct volumes of brains stained by TTC were observed 48 hours later. Brain temperature, blood flow and neurological scores were also recorded during this procedure. RESULTS: In the 1.5, 2, 2.5 and 3 hour ischemic groups, cold saline (20 degrees C infusion via the MCA) rapidly reduced the temperature of the MCA-supplied ischemic territory in the cortex from 37.0-37.1 to 32.8-33.2 degrees C and in the striatum from 37.3-37.5 to 33.2-33.3 degrees C. In NT groups, the average total infarct volumes of 1.5 and 2 hour ischemia (29.80 +/- 2.20 and 34.29 +/- 2.14%, respectively) were significantly less than that of the control group (48.41 +/- 5.82%), but the average total infarct volumes of the 2.5 and 3 hour ischemia groups (47.31 +/- 4.72 and 50.17 +/- 8.08%, respectively) did not change. Compared with the ischemia groups without local saline infusion, the average total infarct volumes of 1.5, 2 and 2.5 hours with local saline infusion to the ischemic territory (16.79 +/- 2.51, 23.09 +/- 4.63% and 25.19 +/- 7.82%, respectively) decreased significantly, but the average total infarct volume of 3 hour ischemia with local saline infusion (43.30 +/- 2.62%) was not different. CONCLUSION: Local cold saline infusion to the ischemic territory before reperfusion can lead to mild hypothermia of the ischemic territory and can prolong the therapeutic time window of reperfusion from 2 to 2.5 hours. Refinements of the cooling process, optimal target temperature, duration of the therapy and most importantly, clinical efficacy, require further study.

Induction of G2/M arrest by pseudolaric acid B is mediated by activation of the ATM signaling pathway.

AIM: The aim of this study was to investigate the mechanism of pseudolaric acid B (PLAB)-induced cell cycle arrest in human melanoma SK-28 cells. METHODS: Cell growth inhibition was detected by MTT assay, the cell cycle was analyzed by flow cytometry, and protein expression was examined by Western blot analysis. RESULTS: PLAB inhibited the growth of human melanoma cells and induced G(2)/M arrest in SK-28 cells, accompanied by an up-regulation of Cdc2 phosphorylation and a subsequent down-regulation of Cdc2 expression. Furthermore, PLAB decreased the expression of Cdc25C phosphatase and increased the expression of Wee1 kinase. Meanwhile, a reduction in Cdc2 activity was partly due to induction of the expression of p21(waf1/cip1) in a p53-dependent manner. In addition, PLAB activated the checkpoint kinase, Chk2, and increased the expression of p53, two major targets of ATM kinase. These effects were inhibited by caffeine, an ATM kinase inhibitor. We also found that PLAB significantly enhanced ATM kinase activity. CONCLUSION: Taken together, these results suggest that PLAB induced G(2)/M arrest in human melanoma cells via a mechanism involving the activation of ATM, and the effect of PLAB on Cdc2 activity was mediated via interactions with the Chk2-Cdc25C and p53 signalling pathways, two distinct downstream pathways of ATM. PLAB may be a promising chemopreventive agent for treating human melanoma.