High efficiency removal of NO using waste calcium carbide slag by facile KOH modification.

Waste calcium carbide slags (CS), which are widely applied to desulfurisation, are not typically used in denitration. Herein, to well achieve waste control by waste, a facile and high-efficiency denitration strategy is developed using KOH to modify the calcium carbide slags (KCS). Various KCS samples were investigated using a series of physical and chemical characterisations. The performance test results showed that the KOH concentration and reaction temperature are the main factors affecting the denitration efficiency of KCS, and CS modified with 1.5 mol/L KOH (KCS-1.5) can achieve 100% denitration efficiency at 300°C. Such excellent removal efficiency is due to the catalytic oxidation of the oxygen-containing functional groups derived from the KCS. Further studies showed that KOH treatment significantly increased the concentration of oxygen vacancies, nitro compounds, and basic sites of CS. This study provides a novel strategy for the resource utilisation of waste CS in the future.

Genotyping genetic markers from LCN and degraded DNA by HRM and their application in hair shaft.

Degraded and low copy number (LCN) DNA samples are common challenging materials in forensic casework because they increase the difficulty of sample processing and reduce the possibility of obtaining genetic information from DNA. High-resolution melting (HRM) curve analysis is promising for genotyping genetic markers and has been applied to the detection of LCN and degraded DNA in the field of forensic science. However, the exact assessment based on HRM at multiple genetic markers from both degraded and LCN DNA materials has not been optimized. To explore the ability of HRM to genotype LCN and degraded DNA samples, we selected three genetic markers to genotype in experimental LCN and degraded DNA and practical hair shaft materials, which are often encountered as degraded and LCN DNA in forensic medicine. The results show that DNA samples of as low as 100 pg and as short as 60 bp were successfully genotyped by the HRM assay at all three genetic markers, whereas in hair shaft DNA, two loci were accurately genotyped. The HRM assay established in this study can be applied to LCN and degraded DNA analysis in forensic casework and can act as a reference point before genotyping short tandem repeat markers. Developing the HRM strategy for genotyping DNA genetic markers enriches detectable targets in hair shaft samples and provides valuable data for further exploration.

High-Resolution Melting Analysis as a Developed Method for Genotyping the PD Susceptibility Loci in LRRK2 Gene.

BACKGROUND: Single-nucleotide polymorphisms (SNPs) have been reported as a highly relevant point for the mechanisms of Parkinson's disease (PD). The invention of saturating dye makes it possible to identify heteroduplex DNA without redistribution during melting, which allows using high-resolution melting (HRM) to detect SNPs. However, the HRM analysis for detection of those SNPs associated with PD was rarely applied. METHODS: Two SNPs, G2385R and R1628P, located in leucine-rich repeat kinase 2 (LRRK2) gene were individually and multiplexedly genotyped using HRM analysis. The sequence variant observed in unexpected HRM curves was confirmed by DNA sequencing. RESULTS: HRM analysis identified successfully all genotypes both on R1628P and G2385R loci. The unexpected HRM curves appeared in R1628P amplicon generated from combinations of R1628P and rs11176013 loci. A multiplexed HRM assay that genotyped R1628P, rs11176013, and G2385R loci was efficiently established. CONCLUSIONS: The present HRM assay is a reliable and rapid method for genotyping R1628P and G2385R loci in LRRK2 gene, and multiplex HRM analysis results in high throughput and has the potential to facilitate a wide range of genotyping studies on PD susceptibility genes.

Proteomic characterization of secretory granules in dopaminergic neurons indicates chromogranin/secretogranin-mediated protein processing impairment in Parkinson's disease.

Parkinson's disease (PD) is an aging disorder related to vesicle transport dysfunctions and neurotransmitter secretion. Secretory granules (SGs) are large dense-core vesicles for the biosynthesis of neuropeptides and hormones. At present, the involvement of SGs impairment in PD remains unclear. In the current study, we found that the number of SGs in tyrosine hydroxylase-positive neurons and the marker proteins secretogranin III (Scg3) significantly decreased in the substantia nigra and striatum regions of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) exposed mice. Proteomic study of SGs purified from the dopaminergic SH-sy5Y cells under 1-methyl-4-phenylpyridinium (MPP+) treatments (ProteomeXchange PXD023937) identified 536 significantly differentially expressed proteins. The result indicated that disabled lysosome and peroxisome, lipid and energy metabolism disorders are three characteristic features. Protein-protein interaction analysis of 56 secretory proteins and 140 secreted proteins suggested that the peptide processing mediated by chromogranin/secretogranin in SGs was remarkably compromised, accompanied by decreased candidate proteins and peptides neurosecretory protein (VGF), neuropeptide Y, apolipoprotein E, and an increased level of proenkephalin. The current study provided an extensive proteinogram of SGs in PD. It is helpful to understand the molecular mechanisms in the disease.

CircSLC8A1 and circNFIX can be used as auxiliary diagnostic markers for sudden cardiac death caused by acute ischemic heart disease.

Sudden cardiac death (SCD) caused by acute ischemic heart disease (IHD) is a major cause of sudden death worldwide. Circular RNAs (circRNAs) are abundant in the heart and play important roles in cardiovascular diseases, but the role of circRNAs as biomarkers in the forensic diagnosis of SCD caused by acute IHD remains poorly characterized. To investigate the potential of two heart-enriched circRNAs, circNFIX and circSLC8A1, we explored the expression of these two circRNAs in different kinds of commonly used IHD models, and further verified their expressions in forensic autopsy cases. The results from both the IHD rat and H9c2 cell models revealed that circSlc8a1 level was upregulated, while the circNfix level was elevated in the early stage of ischemia and subsequently downregulated. The time-dependent expression patterns of the two circRNAs suggested their potential as SCD biomarkers. In autopsy cases, the results showed that the expression of these two circRNAs in the myocardium with acute IHD-related SCDs corresponded to the observations in the ischemic models. Further analysis related to myocardial ischemia indicated that circSLC8A1 showed high sensitivity and specificity for myocardial infarction and was positively correlated with creatine kinase MB in pericardial fluid. Downregulated circNFIX level could indicate the ischemic myocardial damage, and it was negatively correlated with the coronary artery stenosis grade. The combination of circSLC8A1 and circNFIX had better performance to discriminate IHD-related SCDs. The results suggested that circSLC8A1 and circNFIX may be used as auxiliary diagnostic markers for SCD caused by acute IHD in forensic medicine.

Secretogranin III upregulation is involved in parkinsonian toxin-mediated astroglia activation.

Environmental neurotoxins such as paraquat (PQ), manganese, and 1-1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) are associated with a higher risk of Parkinson's disease (PD). These parkinsonian toxins exert certain common toxicological effects on astroglia; however, their role in the regulatory functions of astroglial secretory proteins remains unclear. In a previous study, we observed that secretogranin II (SCG2) and secretogranin III (SCG3), which are important components of the regulated secretory pathway, were elevated in PQ-activated U118 astroglia. In the current study, we used the parkinsonian toxins dopamine (DA), active metabolite of MPTP (MPP+), MnCl2, and lipopolysaccharide (LPS) as inducers, and studied the potential regulation of SCG2 and SCG3. Our results showed that all the parkinsonian toxins except LPS affected astroglial viability but did not cause apoptosis. Exposure to DA, MPP+, and MnCl2 upregulated glial fibrillary acidic protein (GFAP), a marker for astrocyte activation, and stimulated the levels of several astrocytic-derived factors. Further, DA, MPP+, and MnCl2 exposure impeded astroglial cell cycle progression. Moreover, the expression of SCG3 was elevated, while its exosecretion was inhibited in astroglia activated by parkinsonian toxins. The level of SCG2 remained unchanged. In combination with our previous findings, the results of this study indicate that SCG3 may act as a cofactor in astrocyte activation stimulated by various toxins, and the regulation of SCG3 could be involved in the toxicological mechanism by which parkinsonian toxins affect astroglia.

Synthesis and activity evaluation of phenylurea derivatives as potent antitumor agents.

We have discovered several tubulin-active compounds in our previous studies. In the establishment of a compound library of small molecule weight tubulin ligands, 14 new N-3-haloacylaminophenyl-N'-(alkyl/aryl) urea analogs were designed and synthesized. The structure-activity relationship (SAR) analysis revealed that (i) the order of anticancer potency for the 3-haloacylamino chain was following -CH(2)Br>-CHBrCH(3); (ii) the N'-substituent moiety was not essential for the anticancer activity, and a proper alkyl substitution might enhance the anticancer activity. Among these analogs, the compounds 16j bearing bromoacetyl at the N'-end exhibited a potent activity against eight human tumor cell lines, including CEM (leukemia), Daudi (lymphoma), MCF-7 (breast cancer), Bel-7402 (hepatoma), DU-145 (prostate cancer), DND-1A (melanoma), LOVO (colon cancer) and MIA Paca (pancreatic cancer), with the IC(50) values between 0.38 and 4.07 microM. Interestingly, compound 16j killed cancer cells with a mechanism independent of the tubulin-based mechanism, indicating a significant change of the action mode after the structure modification.

Establishment of an alternative efficiently genotyping strategy for human ABO gene.

ABO genotyping is used in several disciplines, including transfusion, transplantation, human evolution, and forensic medicine. Detection of single nucleotide polymorphisms (SNPs) on a locus is a common way to identify different genotypes. In this study we developed a strategy for ABO genotyping, which can rapidly and efficiently detect SNPs. DNA fragments containing 4 SNPs in the ABO gene (c.261delG, c.297A > G, c.1009A > G, and c.1061delC) were amplified using individually and multiplexed polymerase chain reaction (PCR)-based methods and subsequently genotyped by high-resolution melting (HRM) analysis. Human blood ABO genotypes from 92 samples were successfully determined by HRM analysis. A total of 14 genotypes (A/A, A/O01, A/O02, A201/O01, A205/O01, B/B, B/O01, B/O02, A/B, A201/B, A205/B, O01/O01, O02/O02, O01/O02) were identified by analysis of the 4 SNPs of interest in this study. The results suggest that the present HRM assay is a reliable and rapid method for ABO blood type genotyping and it may offer an alternative to traditional genotyping methods.

[Determination of ceftazidime and impurities using high performance liquid chromatography].

A high performance liquid chromatographic (HPLC) method for the determination of ceftazidime and impurities in ceftazidime drug was developed and verified. An Alltima C18 column (250 mm x 4.6 mm, 5 microm) was used as the analysis column. Acetonitrile and phosphate buffer (22.6 g/L aqueous solution of ammonium dihydrogen phosphate, adjusted to pH 3.9 with 10% (v/v) phosphoric acid) were used as mobile phases with gradient elution at a flow rate of 1.3 mL/min. The column temperature was kept at 35 degrees C, and the detection wavelength was set at 255 nm. Fourteen impurities could be well separated. The assay exhibited a good linearity in the ceftazidime concentration range of 0.267-1069 microg/mL with a correlation coefficient of 1.0000. The limits of the quantitation and qualification of ceftazidime were 3.1 ng and 0.93 ng, respectively. The relative standard deviations (RSDs) of the interday and intraday (n=3) determinations at three concentration levels were 0.72% and 0.91%, respectively. At 4 degrees C ang under darkness, ceftazidime solution was stable for 24 h. The developed method is superior to the counterparts in British and Japanese pharmacopeias in the number of the impurities separated and detected.