Electrochemical detection of genetic damage caused by the interaction of novel bifunctional anthraquinone-temozolomide antitumor hybrids with DNA modified electrode.

In this work, novel potential anthraquinone-temozolomide (TMZ) antitumor hybrids N-(2-((9,10-dioxo-9,10-dihydroanthracen-1-yl)amino)ethyl)-3-methyl-4-oxo-3,4-dihydroimidazo [5, 1-d][1,2,3,5]tetrazine-8-carboxamide (C-1) and 2-(9,10-dioxo-9,10-dihydroanthracen-1-yl)amino) ethyl-3-methyl-4-oxo-3,4-dihydroimidazo[5,1-d][1,2,3,5]tetrazine-8-carboxylate (C-9) were designed and synthesized successfully. The electrochemical behaviors of C-1 (C-9) involved the reversible processes of 9,10-anthraquinone ring, the irreversible reduction and oxidation processes of TMZ ring. Electrochemical biosensors were constructed with ctDNA, poly (dG) and poly (dA) modifying the surface of glassy carbon electrode (GCE) to evaluate the DNA oxidative damage caused by the interaction of C-1 (C-9) with DNA. Anthracycline skeleton and TMZ ring in C-1 (C-9) could exhibit bifunctional effects with both intercalating and alkylation modes toward DNA strands. The DNA biosensor had good practicability in mouse serum. The results of gel electrophoresis further demonstrated that C-1 (C-9) could effectively intercalated into ctDNA and disrupt plasmid conformation. Finally, anthraquinone-TMZ hybrid C-1 possessed high cytotoxicity toward A549 and GL261 cells, which could be a novel and optimal candidate for the clinic antitumor treatment.

Electrochemical detection of the oxidative damage of a potential pyrimido[5,4-g]pteridine-derived antitumor agent toward DNA.

In this work, we design and synthesize 2,2'-(7,9-dimethyl-2,4,6,8-tetraoxo-6,7,8,9-tetrahydropyrimido[5,4-g]pteridine-1,3(2H,4H)-diyl)bis(N,N-bis(2-chloroethyl)acetamide) (PT-MCA) as a novel DNA intercalator and potential antitumor agent. Electrochemical analysis reveals the redox process of PT-MCA on the electrode surface. The bioelectrochemical sensors are obtained by modifying the surface of GCE with calf thymus DNA (ctDNA), poly (dG), poly (dA), and G-quadruplex, respectively. The DNA oxidative damage induced by PT-MCA is investigated by comparing the peak intensity change of dGuo and dAdo and monitoring the peaks of the oxidation products of guanine and/or adenine (8-oxoGua and/or 2,8-oxoAde). UV-vis absorption and fluorescence spectra and gel electrophoresis are further employed to understand the intercalation of PT-MCA into DNA base pairs. Moreover, PT-MCA is proved to exhibit stronger anti-proliferation activity than mitoxantrone against both 4T1 and B16-F10 cancer cells. At last, the oxidative damage of PT-MCA toward ctDNA is not interfered by the coexistence of ions and also can be detected in real serums.

A child with household transmitted COVID-19.

BACKGROUND: Although people of all ages are susceptible to the novel coronavirus infection, which is presently named "Coronavirus Disease 2019" (COVID-19), there has been relatively few cases reported among children. Therefore, it is necessary to understand the clinical characteristics of COVID-19 in children and the differences from adults. CASE PRESENTATION: We report one pediatric case of COVID-19. A 14-month-old boy was admitted to the hospital with a symptom of fever, and was diagnosed with a mild form of COVID-19. The child's mother and grandmother also tested positive for SARS-CoV-2 RNA. However, the lymphocyte counts were normal. The chest computed tomography (CT) revealed scattered ground glass opacities in the right lower lobe close to the pleura and resorption after the treatment. The patient continued to test positive for SARS-CoV-2 RNA in the nasopharyngeal swabs and stool at 17 days after the disappearance of symptoms. CONCLUSION: The present pediatric case of COVID-19 was acquired through household transmission, and the symptoms were mild. Lymphocyte counts did not significantly decrease. The RNA of SARS-CoV-2 in stool and nasopharyngeal swabs remained positive for an extended period of time after the disappearance of symptoms. This suggests that attention should be given to the potential contagiousness of pediatric COVID-19 cases after clinical recovery.

[Heavy Metal Pollution and Potential Ecological Risk of Soil from Reclaimed Industrial Sites and Surrounding River Sediments].

Eight heavy metals (As, Cd, Cr, Cu, Ni, Pb, Zn, and Hg) in the soils from 53 reclaimed industrial sites and 23 river sediments in the Shanghai suburbs were determined using the Nemero pollution index and Hakanson potential ecological risk index (RI). The results show that the eight heavy metals exceed the secondary standard limits to different extents. The rate of standard excess is the most prominent for Cu and Zn; it accounts for 47.17% and 43.40%, respectively. The contents of heavy metals in the sediments from surrounding rivers are significantly lower than those in soil from reclaimed industrial sites. The comprehensive Nemero pollution index of heavy metals in reclaimed soil is 11.41, which represents a serious pollution level; Cu and Zn are the most prominent heavy metals. The Nemero pollution index of heavy metals in surrounding river sediments is 1.26, indicating slight pollution. Based on the single-potential ecological risk, the heavy metals at the reclaimed industrial sites all have a slightly latent ecological risk level, except for Cd (strong), Hg (strong), and Cu (moderate). The potential ecological RI is high (385.79). Heavy metals from the surrounding river sediments have a mild potential ecological risk level, except for Hg. Overall, the potential ecological level is mild (RI=83.91). In short, Cu and Zn are more prominent in reclaimed industrial soil. With respect to the toxic response factor, Cd and Hg should be focused on. The heavy metal pollution of surrounding river sediments is light, leading to a mild potential ecological risk level.

Selective Hydrodeoxygenation of 5-Hydroxymethylfurfural to 2,5-Dimethylfuran over Heterogeneous Iron Catalysts.

This work provided the first example of selective hydrodeoxygenation of 5-hydroxymethylfurfural (HMF) to 2,5-dimethylfuran (DMF) over heterogeneous Fe catalysts. A catalyst prepared by the pyrolysis of an Fe-phenanthroline complex on activated carbon at 800 °C was demonstrated to be the most active heterogeneous Fe catalyst. Under the optimal reaction conditions, complete conversion of HMF was achieved with 86.2 % selectivity to DMF. The reaction pathway was investigated thoroughly, and the hydrogenation of the C=O bond in HMF was demonstrated to be the rate-determining step during the hydrodeoxygenation, which could be accelerated greatly by using alcohol solvents as additional H-donors. The excellent stability of the Fe catalyst, which was probably a result of the well-preserved active species and the pore structure of the Fe catalyst in the presence of H2 , was demonstrated in batch and continuous flow fixed-bed reactors.

[Serum carbohydrate antigen-125 levels in patients with bronchiectasis].

OBJECTIVE: To evaluate the level of serum carbohydrate antigen-125(CA125) and its related factors in patients with bronchiectasis. METHODS: The clinical data of 504 patients with bronchiectasis in Zhejiang Putuo People's Hospital from June 2009 to June 2014 were collected in the study.The patients were divided into CA125 elevated group and CA125 normal group according to serum CA125 level,and the differences of serum CA125,age,gender, white blood cell(WBC),C-reactive protein(CRP), blood glucose and other test indicators were compared between two groups. RESULTS: There were 276 patients including 117 male and 159 female with elevated serum CA125.Their mean age was(66.3±13.1)years and the mean level of CA125 was(83.70±43.87) U/mL. There were 228 patients including 84 male and 144 female with normal CA125 levels. Their mean age was(67.5±10.5) years and the mean level of CA125 was(20.68±9.67)U/mL.The peripheral blood WBC in patients with CA125 elevated group[(10.08±5.68)×10(9)/L] was significantly higher than that in CA125 normal group[7.73±3.46)×10(9)/L], the difference was statistically significant(P<0.05).The medium of CRP level in patients with CA125 elevated group[22.98(3.18~196.88)mg/L] was significantly higher than that in CA125 normal group[6.34(0.50~97.66)mg/L](P<0.05). Correlation analysis showed that CA125 was positively correlated with WBC and CRP(P<0.05). Stepwise regression analysis showed that CRP was the only independent prognostic factors of CA125. Paired t test showed the presence of CA125 serum in patients with bronchiectasis had a significant difference between before and after anti-infection therapy(P<0.05). CONCLUSION: The serum levels of CA125 rise in patients with bronchiectasis,while it decrease after anti-infection therapy.CRP is an independent associated factor of serum CA125 level.

Action of thymol on spontaneous excitatory transmission in adult rat spinal substantia gelatinosa neurons.

Thymol, which is contained in thyme essential oil, has various actions including antinociception and nerve conduction inhibition. Although thymol activates transient receptor potential (TRP) channels expressed in heterologous cells, it remains to be examined whether this is so in native neurons. It has not yet been examined how thymol affects synaptic transmission. In order to know how thymol modulates excitatory transmission with a focus on TRP activation, we investigated its effect on glutamatergic spontaneous excitatory transmission in lamina II (substantia gelatinosa; SG) neurons with which nerve terminals expressing TRP channels make synaptic contacts. The experiment was performed by using the blind whole-cell patch-clamp technique in adult rat spinal cord slices. Superfusing thymol (1 mM) for 3 min reversibly increased the frequency of spontaneous excitatory postsynaptic current (sEPSC) with a minimal increase in its amplitude in all neurons examined. Seventy-seven% of the neurons produced an outward current at a holding potential of -70 mV. The sEPSC frequency increase and outward current produced by thymol were concentration-dependent with almost the same half-maximal effective concentration (EC50) values of 0.18 and 0.14 mM, respectively. These activities were repeated at a time interval of 30 min, although the sEPSC frequency increase but not outward current recovered with a slow time course. Voltage-gated Na(+)-channel blocker tetrodotoxin did not affect the thymol activities. The sEPSC frequency increase was inhibited by TRPA1 antagonist HC-030031 but not TRPV1 and TRPM8 antagonist (capsazepine and BCTC, respectively), while these antagonists had no effect on the outward current. This was so, albeit the two thymol activities had similar EC50 values. It is concluded that thymol increases the spontaneous release of L-glutamate onto SG neurons by activating TRPA1 channels while producing an outward current without TRP activation. Considering that the SG plays a pivotal role in modulating nociceptive transmission from the periphery, these actions of thymol could contribute to at least a part of its antinociceptive effect.

Excretion of urinary orosomucoid 1 protein is elevated in patients with chronic heart failure.

Easily screening markers for early detection of chronic heart failure (CHF) are lacking. We identified twenty differently expressed proteins including orosomucoid 1(ORM1) in urine between patients with CHF and normal controls by proteomic methods. Bioinformatics analyses suggested ORM1 could be used for further analysis. After verification by western blotting, the urinary levels of ORM1 were quantified with enzyme-linked immunosorbent assay (ELISA) by correcting for creatinine expression. The ORM1-Cr was significantly elevated in CHF patients than normal controls (6498.83 ± 4300.21 versus 2102.26 ± 1069.24 ng/mg). Furthermore, a Spearman analysis indicated that the urinary ORM1 levels had a high positive correlation with the classification of CHF, and the multivariate analysis suggested that the urinary ORM1 content was associated with the plasma amino-terminal pro- brain natriuretic peptide (NT-proBNP) (OR: 2.106, 95% CI: 1.213-3.524, P = 0.002) and the New York Heart Association (NYHA) classification (OR: 3.019, 95% CI: 1.329-4.721, P<0.001). In addition, receiving operating curve (ROC) analyses suggested that an optimum cut-off value of 2484.98 ng/mg with 90.91% sensitivity and 85.48% specificity, respectively, could be used for the diagnosis of CHF. To sum up, our findings indicate that ORM1 could be a potential novel urinary biomarker for the early detection of CHF.

Increased serum 2-oxoglutarate associated with high myocardial energy expenditure and poor prognosis in chronic heart failure patients.

Myocardial energy expenditure (MEE) and 2-oxoglutarate are elevated in chronic heart failure (CHF) patients compared with healthy controls. To explore whether 2-oxoglutarate could reflect the levels of MEE and predict the prognosis of CHF, 219 CHF patients and 66 healthy controls were enrolled. 2-Oxoglutarate was assayed with Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC/MS/MS). CHF patients were divided into 4 groups according to interquartile range of MEE and followed for death or recurrent hospital admission due to CHF for the mean follow-up time 6.64±0.16months. 2-Oxoglutarate was increased in CHF patients compared with controls (P<0.01) and correlated with estimated glomerular filtration rate (r=0.142, P=0.036), age (r=-0.269, P<0.01) and MEE levels (r=0.307, P<0.01) in a multiple linear correlation analysis in CHF patients. Furthermore, 2-oxoglutarate (OR=3.470, 95% CI=1.557 to 7.730, P=0.002), N-terminal pro-B-type natriuretic peptide (OR=4.013, 95% CI=1.553 to 10.365, P=0.004), age (OR=1.611, 95% CI=1.136 to 2.283, P=0.007) and left ventricular ejection fraction (OR=7.272, 95% CI=3.110 to 17.000, P<0.001) were independently associated with MEE on multiple logistic regression analysis. Kaplan-Meier event curves showed that high 2-oxoglutarate levels were associated with adverse outcomes (Log Rank, Chi(2)=4.026, P=0.045). This study showed that serum 2-oxoglutarate is associated with MEE levels, which can be used as potential biomarkers for MEE, and it can reflect the clinical severity and short-term outcome of CHF.

[Value of quality of life evaluation in prognosis of chronic obstructive pulmonary disease].

OBJECTIVE: To evaluate the quality of life in patients with chronic obstructive pulmonary disease (COPD) by COPD Assessment Test (CAT) and Body Mass Index, Airflow Obstruction, Dyspnea, Exercise Capacity Index (BODE). METHODS: One hundred patients with stable COPD admitted in Putuo People's Hospital were recruited in the study. CAT and BODE index were measured for each patient.The deaths and frequency of exacerbations were recorded during 3-year follow-up period,and the correlation between CAT and BODE in evaluating COPD prognosis was analyzed. RESULTS: There were 28, 30, 29 and 13 patients with CAT score of 1, 2, 3 and 4, respectively; while there were 31, 29, 28 and 12 cases with BODE scores of 1, 2, 3 and 4. CAT scores were well correlated with BODE evaluation in terms of overall score and scores of 4 items (r= -0.237, -0.772, 0.789, -0.767, 0.888, respectively, Ps<0.05). COPD exacerbation incidence and mortality increased with the increasing CAT levels. The rank sum test showed that there were no significant differences between CAT and BODE index in the frequency of acute exacerbation(P<0.05); and in the death toll, the difference was not significant(1 group Χ2=0.919, 2 group Χ2=0.001, 3 group Χ2=0.177,4 group Χ2=0.322, Ps>0.05). CONCLUSION: CAT is relevant to BODE in evaluating incidence of exacerbation and mortality for patients with COPD and CAT is more easily to be applied.

[Research advances on regulation of Pseudomonas aeruginosa biofilm formation and its therapeutic strategies].

Pseudomonas aeruginosa is an important pathogenic bacterium of nosocomial infections. The microbe easily produce biofilm which brings us much difficulties in clinical treatment. The formation processes of biofilm, including the stages of early bacteria planting, mushroom-like structure forming and extracellular matrix producing, are regulated by a series of molecules and genes. And quorum sensing system of the microbe is responsible for regulation of the whole process of biofilm formation. According to the process of biofilm formation and the mimitat associated regulation mechanism, several anti-biofilm therapeutic strategies have been applied in clinical medicine, and some novel drugs and methods are developed.

[The role of p38 mitogen activated protein kinase in the inflammatory response of human monocytes infected with nontypeable Haemophilus influenzae].

OBJECTIVE: To investigate the role of mitogen activated protein kinase (MAPK) pathway in the inflammatory response induced by nontypeable Haemophilus influenzae (NTHi). METHODS: NTHi was a clinical isolate identified by serum agglutination test and 16S rRNA gene sequencing. The peripheral monocytes isolated from adult healthy donors were divided into medium-treated group, NTHi-stimulated group, SB203580 (p38 MAPK inhibitor) -pretreated group and UO126 (p44/42 MAPK inhibitor) -pretreated group. Monocytes were co-cultured with NTHi and harvested 1 h and 4 h after stimulation. The phosphorylation of p38 and p44/42 MAPKs was detected by Western blot. The expression of toll-like receptor 4 (TLR4) was examined by flow cytometry 16 h after bacterial stimulation. Monocytes were preincubated with SB203580 or UO126 for 1 h and then stimulated with NTHi for 4 h and 16 h, respectively. The level of TNF-alpha in the supernatants was determined by Enzyme-linked immunosorbent assay (ELISA). RESULTS: The phosphorylation of p38 MAPK and p44/42 MAPK was rapidly induced by NTHi and continued for at least 4 h after stimulation. The expression of TLR4 on monocytes after NTHi stimulation was significantly up-regulated compared with the control group (11.8 +/- 1.6 vs 4.8 +/- 0.6, P < 0.05). The leve of TNF-alpha in the supernatants was increased 4 h and 16 h after bacterial stimulation compared with the control group (4 h : 16.4 +/- 5.3 vs 0.6 +/- 0.6, P < 0.01; 24 h : 30.2 +/- 10.7 vs 1.4 +/- 1.1, P < 0.01). SB203580 pretreatment decreased remarkably the TNF-alpha secretion from monocytes (P < 0.05) whereas UO126 had no significant effect on TNF-alpha level (P > 0.05). CONCLUSION: TLR4 is probably involved in the inflammatory response of monocytes induced by NTHi whereas p38 MAPK is the key signal molecule in this inflammatory reaction.

A case of chylothorax treated curatively with Sapylin, a streptococcus preparation.

Chylothorax is an uncommon disease where fatty fluid accumulates within the chest cavity. Conservative management, including repeated thoracentesis or pleurodesis, seems to be suitable to most cases. Herein, we present a case of efficacious pleurodesis by intrapleural injection of Sapylin, a streptococcus preparation, for the treatment of chylothorax. A 52-year-old non-smoking female farmer was diagnosed as idiopathic chylothorax after we ruled out possible causes including chest trauma, lymphoma, lung cancer, filariasis, tuberculosis, and etc. Two-time intra-thoracic injection of 3 Klinische Einheit (KE) Sapylin achieved rapid and effective control of chylothorax with no severe side effects. Sapylin may facilitate pleurodesis by producing a strong inflammatory response.