Detection of pathogens from venous or arterial blood of patients with left-sided infective endocarditis by metagenomic next-generation sequencing: A prospective study.

BACKGROUND: Infective endocarditis is a life-threatening uncommon infectious disease, and we aimed to explore the clinical utility of venous or arterial blood-based metagenomic next-generation sequencing (mNGS) approaches to diagnose left-sided infective endocarditis (LSIE). METHODS: We prospectively studied 79 LSIE patients who received valvular surgery in our hospital. Results of blood culture, valve culture, venous blood-based mNGS, arterial blood-based mNGS, venous blood-based mNGS plus blood culture, and arterial blood-based mNGS plus blood culture were evaluated and compared. RESULTS: Both venous blood- and arterial blood-based mNGS methods displayed significantly higher positive detection rates than blood culture and valve culture (43.0 %, 49.4 % vs. 32.9 %, 19.0 %; P < 0.001). Strikingly, when combining blood-based mNGS and blood culture, the positive rate could be further improved to more than 60 %. Moreover, we found mNGS LSIE detection was closely associated with preoperative leukocyte (P = 0.027), neutrophil value (P = 0.018), vegetation ≥ 14 mm (P = 0.043), and vegetations in aortic valve (P = 0.048). In addition, we discovered that blood-based mNGS had a superir capacity over blood culture to detect gram-negative bacteria, fungi, Bartonella Quintana, and mixed infections than blood culture. CONCLUSION: This study indicates that venous blood- and arterial blood-based mNGS displayed high positive rate in the rapid detection of pathogens in high-risk LSIE patients.

Plastic bottles for chilled carbonated beverages as a source of microplastics and nanoplastics.

Carbonated beverages are characterized by low temperatures, multiple microbubbles, high pressure, and an acidic environment, creating ideal conditions for releasing contaminants from plastic bottles. However, the release patterns of microplastics (MPs) and nanoplastics (NPs) are poorly understood. We investigated the effects of plastic type, CO2 filling volume, temperature, sugar content, and additive on the leakage of MPs/NPs and heavy metals. Our results showed that polypropylene bottles released greater MPs (234±9.66 particles/L) and NPs (9.21±0.73 × 107 particles/L) than polyethylene and polyethylene terephthalate bottles. However, subjecting the plastic bottles to 3 repeated inflation treatments resulted in 91.65-93.18% removal of MPs/NPs. The release of MPs/NPs increased with increasing CO2 filling volume, driven by the synergistic effect of CO2 bubbles and pressure. After 4 freeze-thaw cycles, the release of MPs and NPs significantly increased, reaching 450±38.65 MPs and 2.91±0.10 × 108 NPs per liter, respectively. The presence of sugar leads to an elevation in MPs release compared to sucrose-free carbonated water, while the addition of additives to carbonated water exhibits negligible effects on MPs release. Interestingly, actual carbonated beverages demonstrated higher MPs concentrations (260.52±27.18-281.38±61.33 particles/L) than those observed in our well-controlled experimental setup. Our study highlights the non-negligible risk of MPs/NPs in carbonated beverages at low temperatures and suggests strategies to mitigate human ingestion of MPs/NPs, such as selecting appropriate plastic materials, high-pressure carbonated water pretreatment, and minimizing freeze-thaw cycles. Our findings provide insights for further study of the release patterns of the contaminants in natural environments with bubbles, pressure, low temperature, and freeze-thaw conditions.

Application of Metagenomic Next-Generation Sequencing in the Etiological Diagnosis of Infective Endocarditis During the Perioperative Period of Cardiac Surgery: A Prospective Cohort Study.

Objective: The present study aimed to prospectively evaluate the role of metagenomic next-generation sequencing (mNGS) in the etiological diagnosis of patients with perioperative infective endocarditis (IE). Methods: From May 1st, 2019 to December 31st, 2020, a total of 99 patients with IE were enrolled in the present study according to the modified Duke criteria, etiological, and pathological results. 11 non-IE patients undergoing heart valve surgery in the same period were selected as the control group. A blood culture test was performed immediately after admission, and the valves harvested operatively were examined by blood culture and mNGS. Results: In the IE group, there were 29 cases (29.3%) with positive blood culture, 16 cases (16.2%) with positive valve culture, and 85 cases (85.9%) with positive valve mNGS. Compared to culture-based detection, mNGS achieved better performance with a sensitivity, specificity, area under the curve (AUC) of 0.859, 0.727, and 0.793, respectively. The combined approach using culture and mNGS further improved the diagnostic accuracy (sensitivity 89.9%, specificity 72.7%, AUC 0.813). Preoperative white blood cell (P = 0.029) and neutrophils (P = 0.046) were identified as independent factors affecting the detection rate of mNGS. In the mNGS-positive group, 95 strains of pathogens were found and 10 cases were identified with mixed infection. There were 72 gram-positive bacteria and 14 gram-negative bacteria. mNGS positive group displayed higher species richness than mNGS negative group with enrichment of Streptococcus sanguis, Streptococcus buccalis, and Streptococcus griseus. Proteobacteria and Actinomycetes were enriched in mNGS negative group. Notably, six patients showed disconcordant results between culture and mNGS. Rothia aeria was identified in the blood culture, valve culture, and valve mNGS in one patient. Bartonella Quintana and Coxiella burnetii, which were fastidious intracellular bacteria, were found in two blood and valve culture-negative cases. Conclusions: mNGS outperformed the conventional culture method and displayed high accuracy in detecting pathogens in IE patients. This study provided support for the use of mNGS in the etiological diagnosis of IE.

The Research Progress of Trastuzumab-Induced Cardiotoxicity in HER-2-Positive Breast Cancer Treatment.

In recent years, the incidence of breast cancer has been increasing on an annual basis. Human epidermal growth factor receptor-2 (HER-2) is overexpressed in 15-20% human breast cancers, which is associated with poor prognosis and a high recurrence rate. Trastuzumab is the first humanized monoclonal antibody against HER-2. The most significant adverse effect of trastuzumab is cardiotoxicity, which has become an important factor in limiting the safe use of the drug. Unfortunately, the mechanism causing this cardiotoxicity is still not completely understood, and the use of preventive interventions remains controversial. This article focuses on trastuzumab-induced cardiotoxicity, reviewing the clinical application, potential cardiotoxicity, mechanism and discussing the potential interventions through summarizing related researches over the past tens of years.

Dihydroquercetin Activates AMPK/Nrf2/HO-1 Signaling in Macrophages and Attenuates Inflammation in LPS-Induced Endotoxemic Mice.

Dihydroquercetin (DHQ) is a flavonoid compound known for its anti-oxidant effects. Oxidative stress plays a dominant role in regulating the pathways associated with systemic inflammatory immune activation during endotoxemia. Whether and how DHQ regulates inflammatory responses in endotoxemia remains elusive. Here we show DHQ pretreatment effectively reduced the Ten-day mortality in bacterial endotoxin lipopolyssacharide (LPS)-challenged mice, suppressing LPS-induced inflammatory responses reflected by impaired production of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) in the serum of mice. In Raw 264.7 cells, DHQ pretreatment significantly inhibited the transcriptional upregulation of TNF-α, interferon-γ (IFN-γ), interleukin-10 (IL-10) and toll-like receptor 4 (TLR-4) after LPS stimulation. Additionally, knockdown of heme oxygenase-1 (HO-1), one of the most important DHQ induced antioxidant genes, cancelled the inhibition of DHQ treatment on LPS induced TNF-α, IFN-γ production. Nuclear factor erythroid 2-related factor 2 (Nrf2) expression and AMP-activated protein kinase (AMPK) phosphorylation were both enhanced by DHQ in Raw 264.7 cells, indicating a DHQ induced AMPK/Nrf2/HO-1 signal axis. In conclusion, DHQ pretreatment could protect mice against the inflammation and mortality associated with endotoxemia.

Evolutions of antibiotic resistance genes (ARGs), class 1 integron-integrase (intI1) and potential hosts of ARGs during sludge anaerobic digestion with the iron nanoparticles addition.

In this work, we investigated the impact of iron nanoparticle, including magnetite nanoparticles (Fe3O4 NPs) and nanoscale zero-valent iron (nZVI), on the anaerobic digestion (AD) performance. Moreover, the evolutions of antibiotic resistance genes (ARGs), class 1 integrons-integrase (intI1) and potential hosts of ARGs were also investigated. The optimal addition of Fe3O4 NPs and nZVI to promote methane production was 0.5 g/L and 1 g/L, which led to 22.07% and 23.02% increase in methane yield, respectively. The degradation rate of organic matter was also enhanced with the addition of Fe3O4 NPs or nZVI. The results of high-throughput sequencing showed that the reactors with iron NPs exhibited significant differences in microbial community structure, compared to the reactors with the non­iron NPs. Iron NPs have caused the relative abundance of the dominant bacteria (Proteobacteria, Firmicutes and Actinobacteria) generally decreased, while the dominant archaea (Euryarchaeota) increased in AD sludge. Quantitative PCR results revealed that iron NPs accelerated the reductions in total absolute abundance of ARGs, especially a beta-lactamase resistance encoded gene (blaOXA). Network analysis displayed that the attenuation of ARGs was mainly attributed to the decline of potential hosts (Proteobacteria, Firmicutes and Actinobacteria). Meanwhile, environmental factors (such as pH, soluble chemical oxygen demand and heavy metals) were also strongly correlated with ARGs.

Influence of nanoscale zero-valent iron and magnetite nanoparticles on anaerobic digestion performance and macrolide, aminoglycoside, ß-lactam resistance genes reduction.

The effect of nanoscale zero-valent iron (NZVI) and magnetite nanoparticles (Fe3O4 NPs) on anaerobic digestion (AD) performance was investigated through a series of 100-day semi-continuous mesophilic anaerobic digestions. The results indicated that biogas production had increased by 24.44% and 21.66% with the addition of 0.5 g/L Fe3O4 NPs and 1.0 g/L NZVI, respectively. Besides, the abundance of five widespread antibiotic resistance genes (ARGs) (ermF, ermA, ermT, aac(6')-IB, blaOXA-1) was also studied. The decrease in abundance of aac(6')-IB and blaOXA-1 was observed during the AD process with an average removal rate of 95.69% and 44.82%, respectively. Most of the ARGs, especially ermA and ermT, were less abundant in NZVI group compared with control group. The overall results suggested that the addition of NZVI and Fe3O4 NPs contributed to a better sludge anaerobic digestion performance, and NZVI was beneficial to the removal of some ARGs.

Visible-light-driven photocatalytic degradation of sulfamethazine by surface engineering of carbon nitride：Properties, degradation pathway and mechanisms.

Polymeric carbon nitride semiconductor has been explored as emerging metal-free photocatalyst for solving the energy shortage and environmental issues. However, the efficiency of carbon nitride is still not satisfying. Herein, a facile copolymerization between L-cysteine and dicyandiamide has been applied to forming the modified carbon nitride photocatalysts. The photocatalytic performance was evaluated through degrading sulfamethazine under visible light illumination. The ameliorative structure and tuned energy band result in visible-light adsorption enhancement. In addition, nitrogen vacancies offer more sites to adsorbing molecular oxygen, thereby facilitating the transfer of electrons from carbon nitride to the surface adsorbed oxygen. As a result, the degradation rate of optimized modified carbon nitride sample for sulfamethazine was 0.1062 min-1, which was almost 12 times than that of carbon nitride (0.0086 min-1). Superoxide radicals and holes were mainly responsible for the sulfamethazine photodegradation by modified carbon nitride. Two reaction intermediates/products were observed and identified by high performance liquid chromatography-mass spectrometer, and a possible reaction pathway was proposed. This study provides new insights into the design of highly efficient photocatalyst for other organic pollutants degradation.

Enhanced mesophilic anaerobic digestion of waste sludge with the iron nanoparticles addition and kinetic analysis.

As the functional material, iron nanoparticles effectively promote anaerobic digestion (AD) process, including the hydrolysis-acidification process and the biogas production. In this study, nano zero-valent iron (nZVI) and Fe3O4 nanoparticles (Fe3O4 NPs) were added to AD reactors respectively. The AD process was evaluated by the reactors performances, including pH, biogas yields and compositions, as well as the removal ratio of total solids (TS), volatile solids (VS) and soluble chemical oxygen demand (sCOD). Three models (first-order kinetic model, transfer function model and Cone model) were used to explore the kinetics of AD biogas production. The results showed that adding appropriate dose of nZVI or Fe3O4 NPs enhanced anaerobic digestibility of sludge. The highest cumulative biogas yield of 140.34 L with 0.5 g L-1 nZVI and 137.13 L with 1 g L-1 Fe3O4 NPs were obtained by the 80 days of mesophilic operation, respectively. Cumulative biogas productions of these two reactors were significantly enhanced up to 15.70% and 13.44%. TS removal rates reached >70% in all AD reactors with iron nanoparticles, and the highest sCOD removal rates of nZVI and Fe3O4 NPs digesters on the 80th day were 88.22% and 77.63%, respectively. The results of the three-day fermentation experiment and the kinetic parameters showed that the nZVI or Fe3O4 NPs enhanced the hydrolysis-acidification process of the AD, which eventually promoted biogas production. The Cone model was satisfied with the experimental results, which could be used to evaluate the kinetics of AD with iron nanoparticles more reasonably.

Impact of thyroid function on cystatin C in detecting acute kidney injury: a prospective, observational study.

BACKGROUND: Cystatin C (Cys C) used clinically for detecting early acute kidney injury (AKI) was reported to be associated with thyroid function. Therefore, whether the performance of Cys C is affected by thyroid hormones has raised concern in critically ill patients. This study aimed to investigate the impact of thyroid hormones on the diagnostic and predictive accuracy of Cys C for AKI, and hence optimize the clinical application of Cys C. METHODS: A prospective observational study was conducted in the general intensive care units (ICUs). Serum creatinine (SCr), Cys C, and thyroid function were documented for all patients at ICU admission. Patients were separated into five quintiles based on free triiodothyronine (FT3) and total triiodothyronine (TT3), and two categories according to the presence of low T3 syndrome or not. The impact of thyroid function on the performance of Cys C in diagnosing and predicting AKI was assessed by area under the receiver operating characteristic curve (AUC). RESULTS: The AKI incidence was 30.0% (402/1339); 225 patients had AKI upon entry, and 177 patients developed AKI during the subsequent 7 days. The AUCs for Cys C in detecting total AKI, established AKI, and later-onset AKI was 0.753, 0.797, and 0.669, respectively. The multiple linear regression analysis demonstrated that TT3 and FT3 were independently associated with Cys C. Overall, although Cys C did not yield any significant difference in AUCs for detecting AKI among patients with different thyroid hormones, the optimal cut-off value of Cys C to detect AKI was markedly different between patients with and without low T3 syndrome. CONCLUSIONS: The thyroid function had no significant impact on the diagnostic and predictive accuracy of Cys C in detecting AKI in ICU patients. However, the optimal cut-off value of Cys C to detect AKI could be affected by thyroid function.

Triterpenes and Meroterpenes with Neuroprotective Effects from Ganoderma leucocontextum.

Ganoderma leucocontextum is a well-known medicinal mushroom cultivated in the Tibet Plateau of China. Chemistry investigation on the fruiting bodies of this mushroom resulted in the isolation of sixteen secondary metabolites including three new lanostane triterpenes, ganoleucoins Q - S (1 - 3), as well as thirteen known compounds (4 - 16). The structures of compounds 1 - 3 were determined by NMR, MS, CD spectral analysis, and chemical derivation method. The neuroprotective effects of compounds 1 - 16 were tested on PC12 cells. Compounds 1 and 2 showed protective effects against the H2 O2 induced damage with the survival rate of 83.19 ± 0.92%, 73.37 ± 1.25% at the concentration of 200 µm, respectively. Meanwhile, compounds 1 and 2 induced neurite outgrowth at 50 - 200 µm. The results from this study suggested that G. leucocontextum and its metabolites may be potential functional food ingredients for the prevention of neurodegenerative diseases.

Adsorption of phosphate from aqueous solution using iron-zirconium modified activated carbon nanofiber: Performance and mechanism.

Phosphate (P) removal is significant for the prevention of eutrophication in natural waters. In this paper, a novel adsorbent for the removal of P from aqueous solution was synthesized by loading zirconium oxide and iron oxide onto activated carbon nanofiber (ACF-ZrFe) simultaneously. The adsorbent was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The results showed that P adsorption was highly pH dependent and the optimum pH was found to be 4.0. The isotherm of adsorption could be well described by the Langmuir model and the maximum P adsorption capacity was estimated to be 26.3mgP/g at 25°C. The kinetic data were well fitted to the pseudo-second-order equation, indicating that chemical sorption was the rate-limiting step. Moreover, co-existing ions including sulfate (SO42-), chloride (Cl-), nitrate (NO3-) and fluoride (F-) exhibited a distinct effect on P adsorption with the order of F->NO3->Cl->SO42-. Further investigations by FT-IR spectroscopy and pH variations associated with the adsorption process revealed that ligands exchange and electrostatic interactions were the dominant mechanisms for P adsorption. The findings reported in this work highlight the potential of using ACF-ZrFe as an effective adsorbent for the removal of P in natural waters.

Study and health risk assessment of the occurrence of iron and manganese in groundwater at the terminal of the Xiangjiang River.

The pollution of the surface water in the Xiangjiang watershed in China has received much attention, whereas the groundwater conditions in the area have long been ignored. This study investigates the occurrence of Fe and Mn in the groundwater of Chengxi Town located at the terminal of the Xiangjiang River. The study area was monitored for an entire year. Thereafter, the associated health risks were identified. Results showed that approximately 81 and 73 % of the measured samples exceeded the limits for Fe and Mn in Chinese drinking water, respectively. pH level was found to be negatively correlated with the concentrations of Fe and Mn in the groundwater in the study area. The occurrence of Fe in the groundwater showed significant seasonal fluctuations and was possibly affected by the change in environment conditions within the aquifer. By contrast, Mn remained relatively stable in most of the area during the whole year. Overall, no health threats for adults and children in the study area were determined according to the low health index values. Nevertheless, research attention and the implementation of relevant measures are needed for certain villages with exceptionally high Mn concentrations in the groundwater.

Pretreatment of Adipose Derived Stem Cells with Curcumin Facilitates Myocardial Recovery via Antiapoptosis and Angiogenesis.

The poor survival rate of transplanted stem cells in ischemic myocardium has limited their therapeutic efficacy. Curcumin has potent antioxidant property. This study investigates whether prior curcumin treatment protects stem cells from oxidative stress injury and improves myocardial recovery following cells transplantation. Autologous Sprague-Dawley rat adipose derived mesenchymal stem cells (ADSCs) were pretreated with or without curcumin. The hydrogen peroxide/serum deprivation (H2O2/SD) medium was used to mimic the ischemic condition in vitro. Cytoprotective effects of curcumin on ADSCs were evaluated. Curcumin pretreatment significantly increased cell viability and VEGF secretion, and decreased cell injury and apoptosis via regulation of PTEN/Akt/p53 and HO-1 signal proteins expression. The therapeutic potential of ADSCs implantation was investigated in myocardial ischemia-reperfusion injury (IRI) model. Transplantation of curcumin pretreated ADSCs not only resulted in better heart function, higher cells retention, and smaller infarct size, but also decreased myocardial apoptosis, promoted neovascularization, and increased VEGF level in ischemic myocardium. Together, priming of ADSCs with curcumin improved tolerance to oxidative stress injury and resulted in enhancement of their therapeutic potential of ADSCs for myocardial repair. Curcumin pretreatment is a promising adjuvant strategy for stem cells transplantation in myocardial restoration.

Advanced oxidation protein products as prognostic biomarkers for recovery from acute kidney injury after coronary artery bypass grafting.

Recovery from acute kidney injury (AKI) is related to long-term prognosis. This study, involving 56 patients with AKI and 56 controls from a prospective cohort undergoing coronary artery bypass grafting (CABG), investigated the prognostic performance of serum levels of advanced oxidation protein products (AOPPs) for predicting non-recovered AKI and non-completely recovered AKI. AOPP levels increased significantly 7 days after surgery in patients with non-recovered or non-completely recovered AKI. Increased AOPP levels were associated with both types of poor recovery from AKI. Results from receiver-operating characteristic (ROC) curves demonstrated that AOPP levels had good prognostic value for predicting non-recovered and non-completely recovered AKI.

Effects of leukocyte depletion on cardiopulmonary protection and inflammation after valve surgery.

BACKGROUND: To evaluate the effects of leukocyte-depleting filtration on myocardial and pulmonary protection and the inflammatory response in patients undergoing valve surgery. METHODS: Fifty-two patients who underwent mitral valve or mitral and aortic valve replacement were randomized into two groups with or without a leukocyte-depleting filter during surgery. The filter was used from 10 minutes before the release of the aortic cross-clamp to the end of cardiopulmonary bypass. RESULTS: Total leukocyte and neutrophil counts showed a short-term reduction in patients undergoing leukocyte filtration, but there was no significant difference between the two groups during the study. Serum levels of cardiac troponin I were lower than that of the control group (p=0.030). Leukocyte depletion resulted in a significantly higher oxygenation index (p=0.002) and a lower respiratory index (p=0.003) compared with the control group. Serum levels of interleukin-8 were significantly elevated in patients undergoing leukocyte filtration compared with patients without leukocyte filtration (p=0.001). There were no statistically significant differences between the two groups with regards to the concentration of interleukin-6 and TNFα, or the duration of intensive care and hospital stay. CONCLUSIONS: Leukocyte depletion is associated with improved myocardial and lung protection but does not appear to attenuate the inflammatory response in valve surgery.

[Prospective study of cystatin C for diagnosis of acute kidney injury after cardiac surgery].

OBJECTIVE: To prospectively study the value of cystatin C in diagnosis of acute kidney injury (AKI) in patients after cardiac surgery. METHODS: A total of 132 patients undergoing cardiopulmonary bypass were enrolled in this prospectively study. From each patient, blood samples were collected everyday before and after operation to detect the serum creatinine (Scr) and cystatin C levels by enzymatic method and particle-enhanced turbidimetric immunoassay (PETIA), respectively, and the glomerular filtration rate (eGFR) was estimated using MDRD equation. AKI diagnosis was made according to the RIFLE criteria of the Acute Dialysis Quality Initiative (ADQI) (R: Scr increased by > or =50%; I: Scr increased by > or =100%; F: Scr increased by > or =200%; L: Loss of kidney function; E: End-stage renal disease). Another AKI diagnostic criterion was also adopted according to the levels of cystatin C increment, namely an increase by > or =50%, > or =100%, and > or =200%. RESULTS: Twenty-nine patients (21.9%) developed AKI of varied severities, including 10 meeting the R-criteria, 12 the I-criteria, 7 the F-criteria, with the other 103 patients without AKI serving as the control group. Cystatin C of the 29 AKI patients was drastically increased in comparison with that of the control group (P<0.001). Significant linear correlation was found between cystatin C and Scr (r=0.732, P<0.001) and between [cystatin C]-1 and estimated GFR (R=0.803, P<0.001). By the two diagnostic criteria based on cystatin C and Scr levels, respectively, the median diagnostic time of AKI was 2 days (range 1-4 days) and 3 days (range 2-5 days) for R criteria (10 patients, P=0.014), 3.5 days (range 1-6 days) and 5 days (range 2-8 days) for I criteria (12 patients, P=0.008), and 5 days (range 3-7 days) and 6.5 days (range 4-9 days) for F criteria (7 patients, P=0.02), respectively. ROC analysis confirmed excellent accuracy of cystatin C in AKI diagnosis (AUC=0.992). With the cut-off value of cystatin C increment by > or =50%, the diagnostic sensitivity and specificity of AKI was 92% and 95%, respectively. CONCLUSION: Cystatin C can serve as a good indicator for AKI diagnosis to allow earlier detection of AKI than Scr-based diagnosis in patients after cardiac surgery.

[Treatment with oral sildenafil in patients with severe pulmonary hypertension after open heart operations].

OBJECTIVE: To evaluate the clinical effects of treatment with oral sildenafil on severe pulmonary hypertension after cardiac surgery. METHODS: From September 2002 to January 2005, oral sildenafil was added to the treatment regime in 27 cases of severe pulmonary hypertension after cardiac surgery. All these cases were given general treatments including intravenous prostaglandin E1 and inhalation of nitric oxide before the use of sildenafil, which did not show obvious effects on decreasing pulmonary pressure. Then a combined treatment [general treatment plus oral sildenafil (1-2 mg/kg, q8h; Pfizer Ltd)] was instituted. Pulmonary artery pressure, systolic pulmonary artery pressure/systolic systemic blood pressure (Pp/Ps) were measured before and every hour after adding sildenafil. RESULTS: One hour after adding sildenafil, the patients' pulmonary artery pressure decreased remarkably (P < 0.01) with no adverse effects on systematic artery pressure. SO(2) and PaO(2) of all cases improved respectively (P < 0.05). One or two days later, the patients' hemodynamics were stable and some patients stopped inhaling nitric oxide and the dosage of prostaglandin E1 decreased. 25 cases stopped use of ventilator and were discharged safely. 2 cases died of multiple organ dysfunction. CONCLUSION: Sildenafil is a highly selective and effective pulmonary hypertension vasodilator, which can be given for the treatment of pulmonary hypertension after cardiac surgery.

Human multidrug resistance associated protein 4 confers resistance to camptothecins.

PURPOSE: The multidrug resistance associated protein (MRP) 4 is a member of the adenosine triphosphate (ATP)-binding cassette transporter family. Camptothecins (CPTs) have shown substantial anticancer activity against a broad spectrum of tumors by inhibiting DNA topoisomerase I, but tumor resistance is one of the major reasons for therapeutic failure. P-glycoprotein, breast cancer resistance protein, MRP1, and MRP2 have been implicated in resistance to various CPTs including CPT-11 (irinotecan), SN-38 (the active metabolite of CPT-11), and topotecan. In this study, we explored the resistance profiles and intracellular accumulation of a panel of CPTs including CPT, CPT-11, SN-38, rubitecan, and 10-hydroxy-CPT (10-OH-CPT) in HepG2 cells with stably overexpressed human MRP4. Other anticancer agents such as paclitaxel, cyclophosphamide, and carboplatin were also included. METHODS: HepG2 cells were transfected with an empty vehicle plasmid (V/HepG2) or human MRP4 (MRP4/HepG2). The resistance profiles of test drugs in exponentially growing V/HepG2 and MRP4/HepG2 cells were examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazonium bromide (MTT) assay with 4 or 48 h exposure time of the test drug in the absence or presence of various MRP4 inhibitors. The accumulation of CPT-11, SN-38, and paclitaxel by V/HepG2 and MRP4/HepG2 cells was determined by validated high-performance liquid chromatography methods. RESULTS: Based on the resistance folds from the MTT assay with 48 h exposure time of the test drug, MRP4 conferred resistance to CPTs tested in the order 10-OH-CPT (14.21) > SN-38 carboxylate (9.70) > rubitecan (9.06) > SN-38 lactone (8.91) > CPT lactone (7.33) > CPT-11 lactone (5.64) > CPT carboxylate (4.30) > CPT-11 carboxylate (2.68). Overall, overexpression of MRP4 increased the IC50 values 1.78- to 14.21-fold for various CPTs in lactone or carboxylate form. The resistance of MRP4 to various CPTs tested was significantly reversed in the presence of dl-buthionine-(S,R)-sulfoximine (BSO, a gamma-glutamylcysteine synthetase inhibitor), MK571, celecoxib, or diclofenac (all MRP4 inhibitors). In addition, the accumulation of CPT-11 and SN-38 over 120 min in MRP4/HepG2 cells was significantly reduced compared to V/HepG2 cells, whereas the addition of celecoxib, MK571, or BSO significantly increased their accumulation in MRP4/HepG2 cells. There was no significant difference in the intracellular accumulation of paclitaxel in V/HepG2 and MRP4/HepG2 cells, indicating that P-glycoprotein was not involved in the observed resistance to CPTs in this study. MRP4 also conferred resistance to cyclophosphamide and this was partially reversed by BSO. However, MRP4 did not increase resistance to paclitaxel, carboplatin, etoposide (VP-16), 5-fluorouracil, and cyclosporine. CONCLUSIONS: Human MRP4 rendered significant resistance to cyclophosphamide, CPT, CPT-11, SN-38, rubitecan, and 10-OH-CPT. CPT-11 and SN-38 are substrates for MRP4. Further studies are needed to explore the role of MRP4 in resistance, toxicity, and pharmacokinetics of CPTs and cyclophosphamide.