[Application of chromosomal microarray analysis for fetuses with talipes equinovarus].

OBJECTIVE: To investigate the application of fetuses with talipes equinovarus(TE)using chromosomal microarray analysis(CMA)technology. METHODS: From May 2012 to June 2015, 54 fetuses were found with TE and with or without other structural anomalies by prenatal ultrasound. Karyotyping was taking for them all, and the fetuses with normal karyotypes took another CMA test. The data were analyzed with CHAS software. Finally all the cases were followed up to know about their pregnancy outcomes. RESULTS: One of the 54 cases was detected with abnormal karyotype which was trisomy 18(2%, 1/54). CMA was undertaken to the remaining fetuses, they were divided into 2 groups, including isolated TE group(n= 38)and complex TE group(n=15). The detection rate of clinical significant copy number variations(CNV)by CMA was 11%(6/53), while isolated and complex TE group were 5%(2/38)and 4/15, respectively(P= 0.047). Of the 53 cases, 51 cases were successfully followed up. Eleven cases were found without TE after birth, and the false positive rate(FPR)of TE was 22%(11/51). CONCLUSIONS: Whole-genome high-resolution CMA increased the detection rate by 11% in fetuses with TE. With the FPR and the detection rate of the clinical significant CNV of 2 groups, whole-genome CMA could be recommended to the fetuses with complex TE group but normal karyotypes. A series of ultrasonic tests should be suggested to the isolate TE group, while with the abnormal ultrasound, fetuses would be suggested to have CMA test for decreasing the rates of invasive prenatal diagnosis and FPR.

Platelet parameters and expression of platelet membrane glycoprotein in childhood acute lymphoblastic leukemia.

Platelet activation and functional changes were investigated in acute lymphoblastic leukemia (ALL) to provide the basis for early diagnosis and evaluation of curative effect. Platelet parameters, immature platelet fraction (IPF%), immature platelet count (IPC), P-selectin (CD62p) (PAC-1) expression were detected in ALL, ALL-complete remission-induced (CR1), and normal groups with an automatic blood cell analyzer and flow cytometer. CD62p and PAC-1 were higher in the ALL group before adenosine-5-diphosphate (ADP) activation than in the normal group (P < 0.05); PAC-1 expression was higher and lower in the ALL-CR1 group than in normal and ALL groups (P < 0.05), respectively. CD62p and PAC-1 expression was lower in the ALL group than in the normal group after ADP activation (P < 0.05); PAC-1 expression was lower and higher in the ALL-CR1 group than in normal and ALL groups, respectively (P < 0.05). Platelet count (PLT), mean platelet volume (MPV), platelet hematocrit (PCT), and platelet distribution width (PDW) were lower in the ALL group than in the ALL-CR1 group (P < 0.05). PLT, MPV, and PCT did not differ between the ALL-CR1 group and the normal group (P > 0.05). PDW did not differ statistically among all groups (P > 0.05). IPF% and IPC values were higher and lower in the ALL group than in normal and ALL-CR1 groups (P < 0.05), respectively. These did not differ significantly between the normal group and the ALL-CR1 group (P > 0.05). Therefore, ALL patients demonstrate platelet activation and platelet dysfunction; platelet parameters and membrane glycoprotein expression can be used to evaluate the effect of ALL.

Repression of contexual fear memory induced by isoflurane is accompanied by reduction in histone acetylation and rescued by sodium butyrate.

BACKGROUND: Isoflurane produces amnesia in mice during contextual fear conditioning (CFC) trials. Histone acetylation is a form of chromatin modification involved in the transcriptional regulation underlying memory formation. We investigated whether isoflurane-induced repression of contextual fear memory is related to altered histone acetylation in the hippocampus, and whether it can be rescued by the histone deacetylases inhibitor sodium butyrate (SB). METHODS: Adult C57BL/6 mice were chronically given intraperitoneal injections of SB or vehicle for 28 days. Immediately before CFC training, the mice were exposed to isoflurane or air for 30 min and CFC testing was performed the next day. Hippocampal histone acetylation was analysed 1 h after CFC training. c-Fos, an immediate early gene (IEG) suggested to participate in learning and memory formation, was also investigated at the same timepoint. RESULTS: Mice exposed to isoflurane showed a reduction in freezing time during the CFC test. These mice also exhibited reduced hippocampal H3K14, H4K5, and H4K12 acetylation 1 h after CFC training, and also decreased c-Fos expression. All of these changes were attenuated in isoflurane-exposed mice that were chronically treated with SB. CONCLUSIONS: Isoflurane suppresses histone acetylation and down-regulates c-Fos gene expression in CA1 of the hippocampus after CFC training. These changes are associated with isoflurane-induced amnesia. The HDAC inhibitor SB prevented repressed contextual fear memory, presumably by promoting histone acetylation and histone acetylation-mediated gene expression in response to CFC training.

Propagation characteristics of a focused laser beam in a strontium barium niobate photorefractive crystal under reverse external electric field.

The propagation characteristics of a focused laser beam in a SBN:75 photorefractive crystal strongly depend on the signal-to-background intensity ratio (R=Is/Ib) under reverse external electric field. In the range 20>R>0.05, the laser beam shows enhanced self-defocusing behavior with increasing external electric field, while it shows self-focusing in the range 0.03>R>0.01. Spatial solitons are observed under a suitable reverse external electric field for R=0.025. A theoretical model is proposed to explain the experimental observations, which suggest a new type of soliton formation due to "enhancement" not "screening" of the external electrical field.

Isolation and expression analysis of the soybean GmPic gene.

The differential screening method was used to isolate the soy photoperiodic response-related genes and to further elucidate the molecular mechanisms of the soybean photoperiodic response. The light-sensitive species Zhong Dou 24 was used to receive long-time sunshine, short-time sunshine, and natural sunshine treatment. The cDNA-amplified fragment length polymorphism technique was used to screen the differentially expressed cDNA fragments. The rapid amplification of cDNA end technique was used to isolate the gene. Semi-quantitative reverse transcription polymerase chain reaction analysis was used to analyze the gene expression patterns in different light cycles. The gene had a total length of 983 bp, contained a complete open reading frame that encoded 248 amino acids, and shared homology with the mitochondrial phosphate transporter protein. The expression pattern analysis results showed that this gene was expressed in the early stages of soybean growth and development. The short-time sunshine inhibited its expression, whereas the long-time sunshine enhanced its expression. The differential screening method was used to isolate the soybean mitochondrial phosphate transporter gene. The gene may be used as a negative regulatory factor that is involved in the photoperiodic response of soybean.

Efficacy and safety of HSK3486 for the induction and maintenance of general anesthesia in elective surgical patients: a multicenter, randomized, open-label, propofol-controlled phase 2 clinical trial.

OBJECTIVE: To evaluate the efficacy and safety of HSK3486 for the induction and maintenance of general anesthesia in elective surgical patients, but excluding emergency, cardiothoracic, cerebral and endoscopic sinus cases. PATIENTS AND METHODS: A total of 40 eligible patients were randomly assigned to HSK3486 (n = 30) or propofol (n = 10) dosage groups in a ratio of 3:1. Drugs were administered as a bolus injection of 0.4 mg/kg (HSK3486) or 2.0 mg/kg (propofol) for induction, followed by maintenance infusion with the same anesthetic. An additional 6 non-randomized patients received propofol (2.0 mg/kg) for induction and were given HSK3486 for maintenance. RESULTS: The primary efficacy endpoint - the success rate of anesthesia maintenance - was 100% in the 3 arms. The secondary efficacy endpoints included times from discontinuation of HSK3486 or propofol maintenance to full alertness, respiratory recovery, extubation and reaching the goal of the Aldrete score. Also, the proportion of patients who constantly maintained BIS40-60 or those with a period of BIS40-60 during maintenance anesthesia showed no significant difference in the HSK3486 and propofol groups (all p > 0.05). Patients who received HSK3486 exhibited a higher satisfaction score from anesthesiologists during the induction period (p = 0.024). The occurrence and types of treatment-emergent adverse events were similar among the 3 arms, both with a severity of grade 1 or 2. Drug-related hypotension occurred in 14 (46.7%) and 7 (70.0%) patients treated with HSK3486 and propofol, respectively. CONCLUSIONS: HSK3486 exhibited good efficacy for the induction and maintenance of general anesthesia and was well tolerated by patients who underwent elective surgery.

Effects of ciprofol for the induction of general anesthesia in patients scheduled for elective surgery compared to propofol: a phase 3, multicenter, randomized, double-blind, comparative study.

OBJECTIVE: Ciprofol is a newly developed intravenous sedative-hypnotic drug. The objective of the study was to prove whether ciprofol was non-inferior to propofol for the successful induction of general anesthesia. The ideal post-induction sedation level was assessed by comparing patients' clinical symptoms and their hemodynamic effects in responding to noxious stimuli, mostly tracheal intubation and bispectral index (BIS) alterations following ciprofol/propofol administration. PATIENTS AND METHODS: In this multi-center, randomized, double-blind phase 3 trial, selective surgery patients were randomly assigned in a 1:1 ratio to either ciprofol 0.4 mg/kg (n = 88) or propofol 2.0 mg/kg (n = 88) groups. The primary endpoint was the percentage of patients with successful anesthesia inductions. Secondary endpoints included the times to successful induction of general anesthesia and loss of the eyelash reflex, changes in BIS, as well as safety indicators. RESULTS: The anesthesia induction success rates for both ciprofol 0.4 mg/kg and propofol 2 mg/kg groups were 100.0%, with a 95% CI lower success limit of -4.18% difference between the two groups, indicating that ciprofol was non-inferior to propofol. For secondary outcomes, the average time to successful anesthesia and loss of the eyelash reflex were 0.91 min and 0.80 min for ciprofol and 0.80 min and 0.71 min for propofol, respectively. The pattern of BIS changes with ciprofol was similar to propofol and stable during the anesthesia maintenance period. Safety was comparable with 88.6% TEAEs in the ciprofol group compared to 95.5% in the propofol group. The incidence of injection pain was significantly lower in the ciprofol group compared to the propofol group (6.8% vs. 20.5%, p < 0.05). In addition, the patients treated with ciprofol had a lesser increase in blood pressure and heart rate, and fewer cases with BIS > 60 within 15 min of intravenous administration, which indicated that ciprofol may provide a better ideal sedation level during the post-induction period under an equivalent dosing regimen to propofol. CONCLUSIONS: Ciprofol for patients undergoing selective surgery is a new option for the induction of general anesthesia.

Molecular cloning of rhamnose-binding lectin gene and its promoter region from snakehead Channa argus.

Lectins are sugar-binding proteins that mediate pathogen recognition and cell-cell interactions. A rhamnose-binding lectin (RBL) gene and its promoter region have been cloned and characterized from snakehead Channa argus. From the transcription initiation site, snakehead rhamnose-binding lectin (SHL) gene extends 2,382 bp to the end of the 3' untranslated region (UTR), and contains nine exons and eight introns. The open reading frame (ORF) of the SHL transcript has 675 bp which encodes 224 amino acids. The molecular structure of SHL is composed of two tandem repeat carbohydrate recognition domains (CRD) with 35% internal identity. Analysis of the gene organization of SHL indicates that the ancestral gene of RBL may diverge and evolve by exon shuffling and gene duplication, producing new forms to play their own roles in various organisms. The characteristics of SHL gene 5' flanking region are the presence of consensus nuclear factor of interleukin 6 (NF-IL6) and IFN-gamma activation (GAS) sites. The results provide indirect evidence that up-regulation of SHL expression may be induced in response to inflammatory stimuli, such as lipopolysaccharide (LPS), interleukin 6 (IL-6), and interferon gamma (IFN-gamma). The transcript of SHL mRNA was expressed in the head kidney, posterior kidney, spleen, liver, intestine, heart, muscle, and ovary. No tissue-specific expressive pattern is different from reported STLs, WCLs, and PFLs, suggesting that different types of RBLs exist in species-specific fish that have evolved and adapted to their surroundings.

Comparative responses in rare minnow exposed to 17beta-estradiol during different life stages.

Present in the excrement of humans and animals, 17beta-estradiol (E(2)) has been detected in the aquatic environment in a range from several nanograms to several hundred nanograms per liter. In this study, the sensitivities of rare minnows during different life stages to E(2) at environmentally relevant (5, 25, and 100 ng l(-1)) and high (1000 ng l(-1)) concentrations were compared using vitellogenin (VTG) and gonad development as biomarkers under semistatic conditions. After 21 days of exposure, VTG concentrations in whole-body homogenates were analyzed; the results indicated that the lowest observed effective concentration for VTG induction was 25 ng l(-1) E(2) in the adult stage, but 100 ng l(-1) E(2) in the larval and juvenile stages. After exposure in the early life stage, the larval and juvenile fish were transferred to clean water until gonad maturation. No significant difference in VTG induction was found between the exposure and control groups in the adults. However, a markedly increased proportion of females and appearance of hermaphrodism were observed in the juvenile-stage group exposed to 25 ng l(-1) E(2). These results showed that VTG induction in the adult stage is more sensitive than in larval and juvenile stages following exposure to E(2). The juvenile stage may be the critical period of gonad development. Sex ratio could be a sensitive biomarker indicating exposure to xenoestrogens in early-life-stage subchronic exposure tests. The results of this study provide useful information for selecting sensitive biomarkers properly in aquatic toxicology testing.

Structural responses of metallic glasses under neutron irradiation.

Seeking nuclear materials that possess a high resistance to particle irradiation damage is a long-standing issue. Permanent defects, induced by irradiation, are primary structural changes, the accumulation of which will lead to structural damage and performance degradation in crystalline materials served in nuclear plants. In this work, structural responses of neutron irradiation in metallic glasses (MGs) have been investigated by making a series of experimental measurements, coupled with simulations in ZrCu amorphous alloys. It is found that, compared with crystalline alloys, MGs have some specific structural responses to neutron irradiation. Although neutron irradiation can induce transient vacancy-like defects in MGs, they are fully annihilated after structural relaxation by rearrangement of free volumes. In addition, the rearrangement of free volumes depends strongly on constituent elements. In particular, the change in free volumes occurs around the Zr atoms, rather than the Cu centers. This implies that there is a feasible strategy for identifying glassy materials with high structural stability against neutron irradiation by tailoring the microstructures, the systems, or the compositions in alloys. This work will shed light on the development of materials with high irradiation resistance.

The influence of HOXA5-specific siRNA on the expression of Livin and Smac proteins.

OBJECTIVE: To Knockdown Homeobox A5 (HOXA5) expression by HOXA5-specific siRNA and evaluate the effects on Livin and Smac proteins expression in acute T cell leukemia Jurkat cells. MATERIALS AND METHODS: We designed and constructed HOXA5-specific siRNA, and using liposomes we transfected Jurkat cells with this siRNA. The experiment was designed for three groups: (i) experimental group with Jurkat cells transfected with HOXA5-specific siRNA (siRNA transfection group), (ii) negative control group (irrelevant siRNA transfection) with Jurkat cells transfected with pRNAT-U6.1-siD and (iii) normal control group (untransfected Jurkat cells, only with equivalent amounts of cells and medium). We used FQ-PCR and Western blot to detect the relative expression levels of HOXA5 mRNA and protein in each group separately. The Western blot was also used to detect Livin and Smac protein levels in Jurkat cells. RESULTS: Expression levels of HOXA5 mRNA and protein were significantly reduced in the group with Jurkat cells transfected with HOXA5 siRNA (p<0.05). The expression of Livin protein was significantly down-regulated (p<0.05) while the expression of Smac protein was significantly up-regulated (p <0.05). CONCLUSIONS: HOXA5-specific siRNA effectively silenced the HOXA5 gene expression and down-regulation of HOXA5 induced the down-regulation of Livin protein expression and up-regulation of Smac protein. We suggest the HOXA5 gene to be considered as the new target for acute leukemia gene therapy.

Swimming exercise ameliorates neurocognitive impairment induced by neonatal exposure to isoflurane and enhances hippocampal histone acetylation in mice.

Isoflurane-induced neurocognitive impairment in the developing rodent brain is well documented, and regular physical exercise has been demonstrated to be a viable intervention for some types of neurocognitive impairment. This study was designed to investigate the potential protective effect of swimming exercise on both neurocognitive impairment caused by repeated neonatal exposure to isoflurane and the underlying molecular mechanism. Mice received 0.75% isoflurane exposures for 4h on postnatal days 7, 8, and 9. From the third month after anesthesia, the mice were subjected to regular swimming exercise for 4weeks, followed by a contextual fear condition (CFC) trial. We found that repeated neonatal exposure to isoflurane reduced freezing behavior during CFC testing and deregulated hippocampal histone H4K12 acetylation. Conversely, mice subjected to regular swimming exercise showed enhanced hippocampal H3K9, H4K5, and H4K12 acetylation levels, increased numbers of c-Fos-positive cells 1h after CFC training, and less isoflurane-induced memory impairment. We also observed increases in histone acetylation and of cAMP-response element-binding protein (CREB)-binding protein (CBP) during the swimming exercise program. The results suggest that neonatal isoflurane exposure-induced memory impairment was associated with dysregulation of H4K12 acetylation, which may lead to less hippocampal activation following learning tasks. Swimming exercise was associated with enhanced hippocampal histone acetylation and CBP expression. Exercise most likely ameliorated isoflurane-induced memory impairment by enhancing hippocampal histone acetylation and activating more neuron cells during memory formation.

ATRA and As2O3 regulate differentiation of human hematopoietic stem cells into granulocyte progenitor via alteration of HoxB8 expression.

OBJECTIVE: This study aimed to investigate the effect of all-trans retinoic acid (ATRA) and/or arsenic trioxide (As2O3) on homeobox B8 (HOXB8) mRNA and protein expressions during the differentiation and proliferation of hematopoietic stem cells (HSCs) to colony forming unit-granulocyte (CFU-G) in order to explore the pathogenesis of leukemia mediated by HOXB8 at mRNA and protein level. MATERIALS AND METHODS: Twelve cord blood samples were collected from the fetal placenta umbilical vein and cultured in vitro. The proliferation and differentiation of cord blood HSCs into CFU-G was continuously disrupted with 10 nmol/l of ATRA and/or 10 nmol/l of As2O3. The expression of HOXB8 mRNA and protein were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western-blot, respectively. RESULTS: HOXB8 mRNA/protein expression was detected in control, ATRA, As2O3 and ATRA +As2O3 groups on days 3, 7, and 12 of culture. HOXB8 mRNA/protein expression was detectable on day 3, reached its highest level on day 7 and decreased on day 12. HOXB8 mRNA/protein expression in ATRA, As2O3 and ATRA+As2O3 was upregulated compared with control group (p < 0.05). CONCLUSIONS: There is a positive relationship between HOXB8 gene expression and granulocyte progenitor hematopoiesis. ATRA/As2O3 up-regulate the expression of HOXB8 mRNA/protein, and treatment of leukemia with ATRA/As2O3 may regulate HOX gene expression.

Evaluation of preoperative staging in advanced gastric cancer with MRI.

AIM: To evaluate the application of magnetic resonance imaging (MRI) to the preoperative staging of advanced gastric cancer. METHODS: An MRI (SE sequence) was preoperatively performed on 34 patients with advanced gastric cancer. The tumors were located at the cardia fundus in 11 patients, the corpus in 14, the antrum in ten and throughout the entire stomach in two. The images were analyzed and staged on the basis of the criteria proposed by Matsushita M. The results were compared with the corresponding histopathologic findings to analyze the rate of diagnostic accordance. RESULTS: The diagnostic rate accuracy by MRI was 77.8% (seven out of nince) for T2 tumors; 77.3% (17 out of 22) for T3 tumors and 100% (three out of three) for T4 tumors, with the overall accuracy equaling 79.4%. When grades T3 and T4 tumors were considered as a single group to determine the presence or absence of extraserosal invasion using MRI technology, the diagnostic accuracy was 88.3%. Statistically, MRI staging showed a significant correlation with the corresponding histopathologic staging using the Spearman correlation test (rs' = 0.743, P < 0.01). When the concordance between MRI and histopathologic staging results were studied according to tumor location, the staging accuracy was highest (90.9%) in tumors located in gastric cardia fundus. CONCLUSION: MR imaging is moderately valuable when staging advanced gastric cancer, especially for tumors located in gastric cardia fundus.

[Study of the reproducibility and sensitivity of laser microspectral analysis in argon atmosphere at reduced pressure].

In argon atmosphere at reduced pressure, the spectral analysis reproducibility and sensitivity are experimentally studied with the laser microprobe emission spectral analysis(LMESA) system for the metal alloy standard samples. For the atomic spectral analysis line, the relative standard deviation(RSD) in argon atmosphere at reduced pressure is better than that in air atmosphere. For the ionic spectral analysis line, the RSD is almost the same in both cases. The RSD is strongly affected by the composition of the sample and the concentration of elements. The measured slope of calibration curves for the determination of Cu, Zn and Mg in an aluminum alloy in argon atmosphere is about 1.5-2 times of that in the air atmosphere, which indicates an obvious improvement of the sensitivity and is helpful to expand the dynamic range of the spectral analysis.

NF-κB and Nrf2 signaling pathways contribute to wogonin-mediated inhibition of inflammation-associated colorectal carcinogenesis.

The transcriptional factors nuclear factor-κB (NF-κB) and NF-E2-related factor 2 (Nrf2) have been recently reported to have critical roles in protecting various tissues against inflammation and colitis-associated colorectal cancer (aberrant crypt foci). Our previous studies showed that wogonin (5,7-dihydroxy-8-methoxyflavone) possessed anti-neoplastic and anti-inflammatory activities. The present study extended these important earlier findings by exploring the effect of wogonin on the initiation and development of colitis-associated cancer. Wogonin lowered tumor incidence and inhibited the development of colorectal adenomas in azoxymethane- or dextran sulfate sodium-induced mice. We found that wogonin significantly decreased the secretion and expression of IL-6 and IL-1ß, reduced cell proliferation and nuclear expression of NF-κB in adenomas and surrounding tissues and promoted Nrf2 nuclear translocation in surrounding tissues, although overexpressed Nrf2 in tumor tissues was independent of wogonin administration. Furthermore, wogonin inhibited the interaction between human monocytic THP-1 cells and human colon cancer HCT116 cells, and significantly downregulated lipopolysaccharide-induced secretion of prototypical pro-inflammatory cytokines IL-6 and IL-1ß in THP-1 cells. Further mechanism research revealed that wogonin inhibited the nuclear translocation of NF-κB and phosphorylation of IκB and IKKα/ß, and promoted Nrf2 signaling pathway in HCT116 cells and THP-1 cells. Taken together, the present results indicated that wogonin effectively suppressed inflammation-associated colon carcinogenesis and cancer development, suggesting its potential as a chemopreventive agent against colitis-associated colon cancer.

Model for End-Stage Liver Disease (MELD) score does not predict outcomes of hepatitis B-induced acute-on-chronic liver failure in transplant recipients.

BACKGROUND: Acute-on-chronic liver failure (ACLF) is a severe clinical entity and liver transplantation is the only definitive therapy to salvage these patients. However, the timing of liver transplant for these patients remains unclear. METHODS: Seventy-eight patients undergoing liver transplantation because of hepatitis B ACLF were retrospectively analyzed from June 2004 to December 2010. The areas under the receiver operating characteristic curve (AUC) of Model for End-Stage Liver Disease (MELD) score and Child-Turcotte-Pugh (CTP) score for the post-transplantation outcomes were calculated. RESULTS: The median age was 44 years (range, 25-64 years), serum bilirubin 418.53 µmol/L (range, 112.90-971.40 µmol/L), INR 3.177 (range, 1.470-9.850), and creatinine 70.84 µmol/L (range, 12.39-844.1 µmol/L); the median MELD score was 32 (range, 21-53) and CTP score 12 (8-15). The AUCs of MELD and CTP scores for 3-month mortality were 0.581 (95% confidence interval [CI], 0.421-0.742; sensitivity, 87.5%; specificity, 32.8%) and 0.547 (95% CI, 0.401-0.693; sensitivity, 75%; specificity, 41%), respectively. Meanwhile, there were no significant differences in hospital mortality (P = .252) or morbidity (P = .338) between the patients with MELD score ≥30 and those <30. CONCLUSIONS: MELD score had no predictive ability for the outcomes of patients with hepatitis B ACLF after orthotopic liver transplantation.

Tumor necrosis factor-α inhibits angiotensin II receptor type 1 expression in dorsal root ganglion neurons via ß-catenin signaling.

Both tumor necrosis factor (TNF)-α and the angiotensin (Ang) II/angiotensin II receptor type 1 (AT1) axis play important roles in neuropathic pain and nociception. In the present study, we explored the interaction between the two systems by examining the mutual effects between TNF-α and the Ang II/AT1 receptor axis in dorsal root ganglion (DRG) neurons. Rat DRG neurons were treated with TNF-α in different concentrations for different lengths of time in the presence or absence of transcription inhibitor actinomycin D, TNF receptor 1 (TNFR1) inhibitor SPD304, ß-catenin signaling inhibitor CCT031374, or different kinase inhibitors. TNF-α decreased the AT1 receptor mRNA level as well as the AT1a receptor promoter activity in a dose-dependent manner within 30 h, which led to dose-dependent inhibition of Ang II-binding AT1 receptor level on the cell membrane. Actinomycin D (1 mg/ml), SPD304 (50 µM), p38 mitogen-activated protein kinase (MAPK) inhibitor PD169316 (25 µM), and CCT031374 (50 µM) completely abolished the inhibitory effect of TNF-α on AT1 receptor expression. TNF-α dose-dependently increased soluble ß-catenin and phosphorylated GSK-3ß levels, which was blocked by SPD304 and PD169316. In DRG neurons treated with AT2 receptor agonist CGP421140, or Ang II with or without AT1 receptor antagonist losartan or AT2 receptor antagonist PD123319 for 30 h, we found that Ang II and Ang II+PD123319 significantly decreased TNF-α expression, whereas CPG421140 and Ang II+losartan increased TNF-α expression. In conclusion, we demonstrate that TNF-α inhibits AT1 receptor expression at the transcription level via TNFR1 in rat DRG neurons by increasing the soluble ß-catenin level through the p38 MAPK/GSK-3ß pathway. In addition, Ang II appears to inhibit and induce TNF-α expression via the AT1 receptor and the AT2 receptor in DRG neurons, respectively. This is the first evidence of crosstalk between TNF-α and the Ang II/AT receptor axis in DRG neurons.

Oroxylin A induces dissociation of hexokinase II from the mitochondria and inhibits glycolysis by SIRT3-mediated deacetylation of cyclophilin D in breast carcinoma.

Oroxylin A is a major active component of the Chinese traditional medicinal plant Scutellaria baicalensis Georgi, which has been reported as a potential anticancer drug. We demonstrated that, Oroxylin A inhibited the glycolysis and the binding of hexokinase II (HK II) with mitochondria in human breast carcinoma cell lines, which was dependent on sirtuin-3 (SIRT3). The level of SIRT3 in mitochondria was increased by Oroxylin A. Then SIRT3 deacetylated cyclophilin D, diminished its peptidyl-prolyl cis-trans isomerase activity and induced its dissociation from the adenine nucleotide translocator. Finally, SIRT3-induced inactivation of cyclophilin D resulted in the detachment of mitochondrial HK II and the inhibition of glycolysis. These results have important implications for the metabolism reprogramming effect and the susceptibility to Oroxylin A-induced mitochondrial cytotoxicity through the regulation of SIRT3 in breast carcinoma.

Repeated intrathecal administration of ropivacaine causes neurotoxicity in rats.

Previous studies have shown that ropivacaine is the least neurotoxic local anaesthetic. Most of the data derive from short-term ropivacaine injection into the subarachnoid space. Intrathecal administration for a prolonged period, and the histological changes and behavioural effects of repeated intrathecal administration, have not previously been investigated. We studied the possible neurotoxicity of intrathecal injection of ropivacaine in a rat model. Rats received 0.12 ml/kg body weight of ropivacaine at concentrations of 0.5 or 1%, or normal saline only, via an implanted intrathecal catheter at 90-minute intervals for 12 hours. On days 1, 3, 5, 7, 14 and 28, the spinal cord was examined by light and electron microscopy at the L3 level. We assessed sensory thresholds to noxious stimulation, behavioural change and protein kinase B immunoreactivity for possible neuronal injury within the spinal cord. Ropivacaine 1% induced thermal hyperalgesia and mechanical allodynia, neuronal injury characterised by tissue oedema, proliferation of glial cells, neuronal morphology changes and degeneration and protein kinase B expression. There were no significant differences in motor function as a result of different concentrations of ropivacaine. Repeated intrathecal injection of ropivacaine 1% can induce neurotoxicity in rats. Our data suggests that expression of protein kinase B might be involved in this neurotoxicity.