Subclinical paraganglioma of the retroperitoneum: A case report.

BACKGROUND: Paraganglioma (PGL) located in the retroperitoneum presents challenges in diagnosis and treatment due to its hidden location, lack of specific symptoms in the early stages, and absence of distinctive manifestations on imaging. CASE SUMMARY: A 56-year-old woman presented with a left upper abdominal mass discovered 1 wk ago during a physical examination. She did not have a history of smoking, alcohol consumption, or other harmful habits, no surgical procedures or infectious diseases, and had a 4-year history of hypertension. Upon admission, she did not exhibit fever, vomiting, or abdominal distension. Physical examination indicated mild percussion pain in the left upper abdomen, with no palpable enlargement of the liver or spleen. Laboratory tests and tumor markers showed no significant abnormalities. Enhanced computed tomography and magnetic resonance imaging of the upper abdomen revealed a cystic solid mass in the left epigastrium measuring approximately 6.5 cm × 4.5 cm, with inhomogeneous enhancement in the arterial phase, closely associated with the lesser curvature of the stomach and the pancreas. The patient underwent laparoscopic resection of the retroperitoneal mass, which was successfully removed without tumor rupture. A 12-month postoperative follow-up period showed good recovery. CONCLUSION: This case report details the successful laparoscopic resection of a retroperitoneal subclinical PGL, resulting in a good recovery observed at the 12-month follow-up. Interestingly, the patient also experienced unexpected cure of hypertensive disease.

Primary squamous cell carcinoma of the liver: A case report.

BACKGROUND: Squamous cell carcinoma (SCC) of the liver is rare, and is more commonly found in the skin, rectum, cervical or inguinal lymph nodes. CASE SUMMARY: A 73-year-old man had been experiencing right upper quadrant discomfort for some weeks. He had a 50-year history of smoking and drinking. On average, he smoked 20 cigarettes and consumed 200 galcoholdaily. He didn't have a history of hepatitis or surgery. Fever, vomiting, jaundice, dysuria, chills, and abdominal distention were not observed at the time of admission. Tenderness in the right upper quadrant was found on physical examination, but there was no palpable abdominal mass. No obvious abnormalities in laboratory tests and tumor markers were found. The plasma retention rate of indocyanine green (ICG) at 15 min was 1.35%. Subsequent abdominal ultrasonography showed a mixed echoic mass approximately 3.8 cm diameter in the left caudate lobe of the liver. Abdominal computed tomography confirmed a 3.0 cm × 3.5 cm irregular mass with inhomogeneous density and moderate delayed enhancement in the left caudate lobe of the liver. Laparoscopic left caudate lobectomy was performed to remove the liver mass. Intra-operative findings confirmed a non-cirrhotic liver, with a 3 cm × 3.5 cm white tumor mass in the left caudate lobe with no tumor rupture and no hemoperitoneum. The resection margin was 1.0 cm in width. CONCLUSION: We describe the first case of SCC in the left caudate lobe of the liver, which was successfully treated by surgical resection and postoperative immunotherapy. No tumor recurrence was observed during the 8-mo follow-up.

Effects of endoplasmic reticulum stress, liver function, insulin resistance and vascular endothelial function in patients with nonalcoholic fatty liver disease.

It is aimed to compare whether there are differences in endoplasmic reticulum stress, liver function, insulin resistance, and vascular endothelial function in patients with nonalcoholic fatty liver disease (NAFLD) with and without type 2 diabetes mellitus (T2DM). Forty patients with NAFLD, 38 patients with NAFLD combined with T2DM, and 30 patients with normal liver tissue were selected. They were set as Group A, Group B and Group C respectively. The expression level of glucose-regulated protein 94 (GRP94) and biochemical indicators such as alanine aminotransferase (ALT), free fatty acids (FFA), triglycerides (TG), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), fasting blood glucose (FBG), and fasting insulin (FINS) were measured. Also, the calculation of the homeostatic model assessment for insulin resistance (HOMA-IR) index was performed. The reactive hyperemia index (RHI) was detected to evaluate the vascular endothelial function of patients. The comparison between groups and multi-factor analysis of the influencing factors of RHI was conducted. Compared with Group C, the expressions in Group A and Group B were distinctly enhanced (P <0.05). Also, the expression of GRP94 protein in Group B was distinctly higher than that in Group A (P <0.05). The average optical density values of Groups A, B, and C were 0.327 ± 0.007, 0.350 ± 0.009, and 0.299 ± 0.006, respectively. A comparison between the three groups was performed. The differences had statistical significance (P <0.05). The differences in the TG, ALT, AST, GGT, FINS and HOMA-IR between Group A and Group B had statistical significance (P <0.05). The RHI values of Groups A, B, and C were 1.59 ± 0.23, 1.79 ± 0.32, and 2.05 ± 0.47, respectively. A comparison between the three groups was performed. The differences had statistical significance (P <0.05). FFA, ALT and FBG in patients with NAFLD are risk factors for endothelial dysfunction (P <0.05). The liver damage caused by NAFLD may be related to the expression of GRP94. FFA, ALT and FBG are risk factors for endothelial dysfunction in NAFLD patients.

The Incidence and Characteristics of Perinatal Stroke in Beijing: A Multicenter Study.

Objective: To assess the incidence, risk factors, and clinical characteristics of perinatal stroke in Beijing. Methods: This multicenter prospective study included all the live births from 17 representative maternal delivery hospitals in Beijing from March 1, 2019 to February 29, 2020. Neonates with a stroke were assigned to the study group. Clinical data, including general information, clinical manifestations, and risk factors, were collected. Up until 18 months after birth, neonates were routinely assessed according to the Ages and Stages Questionnaire (ASQ) and/or the Bayley scale. Statistical analysis was done using the chi-squared, t-tests, and logistic regression analysis using SPSS version 26.0. Outcomes: In total, 27 cases were identified and the incidence of perinatal stroke in Beijing was 1/2,660 live births, including 1/5,985 for ischemic stroke and 1/4,788 for hemorrhagic stroke. Seventeen cases (62.96%) of acute symptomatic stroke and convulsions within 72 h (10 cases, 37.04%) were the most common presentations. Ten patients showed no neurological symptoms and were found to have had a stroke through routine cranial ultrasonography after being hospitalized for non-neurological diseases. The risk factors include primiparity, placental or uterine abruption/acute chorioamnionitis, intrauterine distress, asphyxia, and severe infection. In the study group, 11.1% (3/27) of patients had adverse neurodevelopmental outcomes. The patients in the study group had lower scores for the ASQ than those in the control group in the communication, gross, and fine motor dimensions. Conclusion: The incidence of perinatal stroke in Beijing was consistent with that in other countries. Routine neuroimaging of infants with risk factors may enable identification of asymptomatic strokes in more patients. Patients who have suffered from a stroke may have neurological sequelae; therefore, early detection, treatment, and regular follow-ups are beneficial for improving their recovery outcomes.

Switching-behavior improvement in HfO2/ZnO bilayer memory devices by tailoring of interfacial and microstructural characteristics.

We investigated the effect of top contact interface and microstructural characteristics of the insulating layers on resistive switching behaviors by fabricating and characterizing the HfO2/ZnO bilayer heterostructures. Different thickness of ZnO underlying layer and different deposition temperatures of the upper HfO2layer were designed to analyze the intrinsic contribution of the crystalline microstructure of the insulating bilayer. Pt and Ti top electrodes were used to demonstrate the extrinsic contribution of the interface configuration. It was observed that all devices show bipolar RS characteristics. Unlike the device composed of Pt/HfO2/ZnO/Pt that exhibit an abrupt switching, a gradually continuous switching in the reset process was identified in the device composed of Ti/HfO2/ZnO/Pt. Interfacial charge migration process/characteristic plays a key role in the RS process as well as its conduction mechanism. The RS performance of the former is significantly better than that of the latter, including much lower reset voltage, two orders of magnitude larger OFF/ON ratio and HRS resistance. In addition, as compared to the intrinsic contribution arising from the microstructure of the HfO2/ZnO bilayer to the RS performances and current transport mechanism, the extrinsic effect contributed from the electrode characteristics (and its interface) is dominant.

Clinical characteristics of 967 children with pertussis: a single-center analysis over an 8-year period in Beijing, China.

The purpose of this study is to understand children's clinical characteristics with pertussis and analyze risk factors on critical pertussis patients. Demographic data from patients with pertussis at Children's Hospital affiliated to the Capital Institute of Pediatrics between March 2011 and December 2018 were collected. We retrospectively gathered more information with the positive exposure, vaccination, antibiotic usage before diagnosis, clinical manifestation, laboratory tests, therapy, and complications for hospitalized children. We divided the patients into severe and non-severe groups, comparing related factors and clinical characteristics among each group. In particular, we summarize the clinical features of the severe patients before aggravation. A total of 967 pertussis cases were diagnosed, of which 227 were hospitalized. The onset age younger than 3 months old accounted for the highest proportion, and 126 patients received hospitalization. For those patients, the incidence of post-tussive vomiting, paroxysmal cyanosis, post-tussive heart rate decrease, hypoxemia, severe pneumonia, and mechanical ventilation was significantly higher than that in the ≥ 3-month-old group (p < 0.05). Among 227 hospitalized patients, 54 suffered from severe pertussis. Risk factors for severe patients included early age of onset, pathogen exposure, and unvaccinated status. Cough paroxysms, post-tussive vomiting, paroxysmal cyanosis, facial flushing/cyanosis/fever during cough, increased WBC, and chest X-ray revealing pneumonia/consolidation/atelectasis were important indications of severe pertussis. Unvaccinated status was an independent risk factor for severe pertussis. The most vulnerable population was infants < 3 months old to pertussis, and may be on the severe end of the disease. Pediatricians must detect and treat severe cases promptly and recommend timely vaccination for all eligible children.

Enhancement of Memory Properties of Pentacene Field-Effect Transistor by the Reconstruction of an Inner Vertical Electric Field with an n-Type Semiconductor Interlayer.

Organic field-effect transistors (OFETs) as nonvolatile memory units are essential for lightweight and flexible electronics, yet the practical application remains a great challenge. The positively charged defects in pentacene film at the interface between pentacene and polymer caused by environmental conditions, as revealed by theoretical and experimental research works, result in unacceptable high programming/erasing (P/E) gate voltages in pentacene OFETs with polymer charge-trapping dielectric. Here, we report a pentacene OFET in which an n-type semiconductor layer was intercalated between a polymer and a blocking insulator. In this structure, the hole barrier caused by the defect layer can be adjusted by the thickness and charge-carrier density of the n-type semiconductor interlayer based on the electrostatic induction theory. This idea was implemented in an OFET structure Cu/pentacene/poly(2-vinyl naphthalene) (PVN)/ZnO/SiO2/Si(p+), which shows low P/E gate voltages, large field-effect mobility (0.73 cm2 V-1 s-1), fast P/E speeds (responding to a pulse width of 5 × 10-4 s), and long retention time in air.

Circulating miRNAs as biomarkers for severe acute pancreatitis associated with acute lung injury.

AIM: To identify circulating micro (mi)RNAs as biological markers for prediction of severe acute pancreatitis (SAP) with acute lung injury (ALI). METHODS: Twenty-four serum samples were respectively collected and classified as SAP associated with ALI and SAP without ALI, and the miRNA expression profiles were determined by microarray analysis. These miRNAs were validated by quantitative reverse transcription-polymerase chain reaction, and their putative targets were predicted by the online software TargetScan, miRanda and PicTar database. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (commonly known as KEGG) were used to predict their possible functions and pathways involved. RESULTS: We investigated 287 miRNAs based on microarray data analysis. Twelve miRNAs were differentially expressed in the patients with SAP with ALI and those with SAP without ALI. Hsa-miR-1260b, 762, 22-3p, 23b and 23a were differently up-regulated and hsa-miR-550a*, 324-5p, 484, 331-3p, 140-3p, 342-3p and 150 were differently down-regulated in patients with SAP with ALI compared to those with SAP without ALI. In addition, 85 putative target genes of the significantly dysregulated miRNAs were found by TargetScan, miRanda and PicTar. Finally, GO and pathway network analysis showed that they were mainly enriched in signal transduction, metabolic processes, cytoplasm and cell membranes. CONCLUSION: This is the first study to identify 12 circulating miRNAs in patients with SAP with ALI, which may be biomarkers for prediction of ALI after SAP.

De novo sequencing and analysis of the cranberry fruit transcriptome to identify putative genes involved in flavonoid biosynthesis, transport and regulation.

BACKGROUND: Cranberries (Vaccinium macrocarpon Ait.), renowned for their excellent health benefits, are an important berry crop. Here, we performed transcriptome sequencing of one cranberry cultivar, from fruits at two different developmental stages, on the Illumina HiSeq 2000 platform. Our main goals were to identify putative genes for major metabolic pathways of bioactive compounds and compare the expression patterns between white fruit (W) and red fruit (R) in cranberry. RESULTS: In this study, two cDNA libraries of W and R were constructed. Approximately 119 million raw sequencing reads were generated and assembled de novo, yielding 57,331 high quality unigenes with an average length of 739 bp. Using BLASTx, 38,460 unigenes were identified as putative homologs of annotated sequences in public protein databases, including NCBI NR, NT, Swiss-Prot, KEGG, COG and GO. Of these, 21,898 unigenes mapped to 128 KEGG pathways, with the metabolic pathways, secondary metabolites, glycerophospholipid metabolism, ether lipid metabolism, starch and sucrose metabolism, purine metabolism, and pyrimidine metabolism being well represented. Among them, many candidate genes were involved in flavonoid biosynthesis, transport and regulation. Furthermore, digital gene expression (DEG) analysis identified 3,257 unigenes that were differentially expressed between the two fruit developmental stages. In addition, 14,473 simple sequence repeats (SSRs) were detected. CONCLUSIONS: Our results present comprehensive gene expression information about the cranberry fruit transcriptome that could facilitate our understanding of the molecular mechanisms of fruit development in cranberries. Although it will be necessary to validate the functions carried out by these genes, these results could be used to improve the quality of breeding programs for the cranberry and related species.

Identification and expression analysis of MATE genes involved in flavonoid transport in blueberry plants.

Multidrug and toxic compound extrusion (MATE) proteins are the most recently identified family of multidrug transporters. In plants, this family is remarkably large compared to the human and bacteria counterpart, highlighting the importance of MATE proteins in this kingdom. Here 33 Unigenes annotated as MATE transporters were found in the blueberry fruit transcriptome, of which eight full-length cDNA sequences were identified and cloned. These proteins are composed of 477-517 residues, with molecular masses ~54 kDa, and theoretical isoelectric points from 5.35 to 8.41. Bioinformatics analysis predicted 10-12 putative transmembrane segments for VcMATEs, and localization to the plasma membrane without an N-terminal signal peptide. All blueberry MATE proteins shared 32.1-84.4% identity, among which VcMATE2, VcMATE3, VcMATE5, VcMATE7, VcMATE8, and VcMATE9 were more similar to the MATE-type flavonoid transporters. Phylogenetic analysis showed VcMATE2, VcMATE3, VcMATE5, VcMATE7, VcMATE8 and VcMATE9 clustered with MATE-type flavonoid transporters, indicating that they might be involved in flavonoid transport. VcMATE1 and VcMATE4 may be involved in the transport of secondary metabolites, the detoxification of xenobiotics, or the export of toxic cations. Real-time quantitative PCR demonstrated that the expression profile of the eight VcMATE genes varied spatially and temporally. Analysis of expression and anthocyanin accumulation indicated that there were some correlation between the expression profile and the accumulation of anthocyanins. These results showed VcMATEs might be involved in diverse physiological functions, and anthocyanins across the membranes might be mutually maintained by MATE-type flavonoid transporters and other mechanisms. This study will enrich the MATE-based transport mechanisms of secondary metabolite, and provide a new biotechonology strategy to develop better nutritional blueberry cultivars.

Plasmonic Ag deposited TiO2 nano-sheet film for enhanced photocatalytic hydrogen production by water splitting.

TiO2 nano-sheet film (TiO2 NSF) was prepared by a hydrothermal method. Ag nanoparticles (NPs) were then deposited on the surface of TiO2 NSF (Ag/TiO2 NSF) under microwave-assisted chemical reduction. The prepared samples were characterized by x-ray diffraction (XRD), scanning electron microscopy (SEM), UV-visible (UV-vis) absorption spectroscopy, x-ray photoelectron spectroscopy (XPS), photoluminescence (PL) spectroscopy, and Raman scattering spectroscopy. The results revealed that the Ag NPs were well dispersed on the anatase/rutile mixed-phase TiO2 nano-sheet surface with a metallic state. The visible light absorption and Raman scattering of TiO2 were enhanced by Ag NPs based on its surface plasmon resonance effect. Besides, Ag NPs could also effectively restrain the recombination of photogenerated electrons and holes. Photocatalytic water splitting was conducted on the films to obtain hydrogen, and the experimental results indicated that plasmonic Ag NPs could greatly enhance the photocatalytic activity of TiO2 due to the synergistic effect between electron transfer and surface plasmon resonance enhanced absorption. The hydrogen yield obtained from the optimal sample reached 8.1 µmol cm(-2) and the corresponding energy efficiency was about 0.47%, which was 8.5 times higher than that of pure TiO2 film. Additionally, the formation mechanism of TiO2 nano-sheet film is preliminarily discussed.

Preparation and characterization of self-assembled percolative BaTiO3-CoFe2O4 nanocomposites via magnetron co-sputtering.

BaTiO3-CoFe2O4 composite films were prepared on (100) SrTiO3 substrates by using a radio-frequency magnetron co-sputtering method at 750 °C. These films contained highly (001)-oriented crystalline phases of perovskite BaTiO3 and spinel CoFe2O4, which can form a self-assembled nanostructure with BaTiO3 well-dispersed into CoFe2O4 under optimized sputtering conditions. A prominent dielectric percolation behavior was observed in the self-assembled nanocomposite. Compared with pure BaTiO3 films sputtered under similar conditions, the nanocomposite film showed higher dielectric constants and lower dielectric losses together with a dramatically suppressed frequency dispersion. This dielectric percolation phenomenon can be explained by the 'micro-capacitor' model, which was supported by measurement results of the electric polarization and leakage current.

[Clinical research of Bordetella pertussis infection in infants with prolonged cough].

OBJECTIVE: To explore the prevalence of Bordetella pertussis (B. pertussis) infection in unvaccinated or incomplete vaccinated infants with cough for a prolonged duration. METHODS: The serum samples and nasopharyngeal secretions were collected from 176 patients with cough for a prolonged duration ( ≥ 2 weeks) from 2011 to 2012 at Children's Hospital Affiliated to Capital Institute of Pediatrics. Multiplex PCR of nasopharyngeal secretion was employed to identify B.pertussis. And enzyme-linked immunosorbent assay(ELISA) was used to detect antibody to pertussis toxin(PT-IgG). Total bacterial DNA was enacted from nasopharyngeal secretion and two-target IS481/PT of B.pertussis was detected by PCR. The sera and nasopharyngeal secretions were also collected from household contacts with cough for a prolonged duration. Their clinical characteristic and epidemiological profiles were collected and analyzed. RESULTS: B.Pertussis infection was demonstrated in 51 cases (29.0%). The patients ages were from 23 days to 4 years. Among them, 46 cases (90.2%) were aged under 12 months and 5 cases (9.8%) over 12 months. And 40 cases were unvaccinated (31 cases <3 months old, 4 cases 3-12 months old, 5 cases >5 years old) and 11 cases incompletely vaccinated. There were 31 males and 20 females. More patients were found in spring and summer than those in autumn and winter. Nine infant cases had 12 household contacts. Among 12 household contacts, 3 were PCR positive and 12 PT-IgG positive. Pertussis was remarkably critical in infants. Serious complications included failure to thrive, pneumonia, respiratory failure and seizures. CONCLUSIONS: B.pertussis infection is an important cause in unvaccinated or incomplete vaccinated infants with prolonged cough. Peak seasons of pertussis are spring and summer. Undiagnosed adolescents and adults with pertussis may be a significant source for transmission of B.pertussis to other susceptible children. Infants aged under 1 year are at risk for severe pertussis and life-threatening complications. As a rapid and sensitive method of detecting B.pertussis, PCR may be used in early phase.

[The experiment research on solution refractive index sensor based on tilted fiber Bragg grating].

The present paper analyzes the sensor's basic principle of the bare tilted fiber Bragg grating (TFBG) and surface plasmon resonance sensor (SPR) that deposited nanoscale gold-coating on the surface of the cladding. We simulated the transmission spectrums and some order cladding mode of TFBG in different concentration solutions by Integration and optical fiber grating software OptiGrating. So by the graphic observation and data analysis, a preliminary conclusion was got that in a certain sensing scope, the cladding modes of TFBG shift slightly to right with the increasing the solution refractive index(SRI),and the relation between resonance peak caused by the coupling of core mode and a certain cladding mode and the SRI was linear. Then the 45 nm thick gold coating was deposited on the surface of the TFBG cladding in a small-scale sputtering chamber KYKY SBC-12, and thermal field scanning electron microscopy presents that the effect of gold-coating was satisfactory to a certain extent in terms of microscopic level. The refractive index(RI) sensing experiments of different concentration solutions of NaCI, MgCI2, CaCI2 were carried out using bare and gold deposited TFBG. The RI sensing characteristics of both bare and gold deposited TFBGs respectively were studied by experiments. Meanwhile, it proved the conclusion that the cladding modes of TFBG drifted to right gradually when the SRI was increasing and the relations between resonance peak caused by the coupling of core mode and a certain cladding mode and the SRI were linear. And by quantitative analysis, we know that SPR sensor with the deposited namoscale gold layer on the surface of cladding enhanced the RI sensitivity dramatically by 2 to 500 nm RIU-1 which is 200 to 300 times larger than that of the bare tilted fiber Bragg grating approximately. The degrees of linear fittings of resonance peak caused by the coupling of core mode and a certain cladding mode and SRI of bare and gold-coating deposited SPR sensor are very good and both of them reach up to more than 0. 99.

[Survey of pertussis infection in infants aged under 3 months with persistent cough].

OBJECTIVE: To explore the prevalence of pertussis in hospitalized infants aged under 3 months with persistent cough. METHODS: The nasopharyngeal secretions and serum samples were collected from hospitalized infants aged under 3 months with cough for over 2 weeks from January 2011 to January 2012. The samples of nasopharyngeal secretion were suctioned and collected. Multiplex PCR assay was employed to identify Bordetella pertussis (B. pertussis) and enzyme-linked immunosorbent assay used to detect antibody to pertussis toxin (PT-IgG). Total bacterial DNA was exacted from nasopharyngeal secretion and two-target IS481/PT of B. pertussis was detected by PCR. RESULTS: Fifty-nine infants (32 boys and 27 girls) were enrolled. None of them was pre-immunized with diphtheria-pertussis-tetanus vaccine. Seventeen infants (28.8%) were B. Pertussis positive. Among 17 cases, 3 infants under 1 month, 4 infants 1 -2 months, and 10 infants 2 - 3 months. Three infants had household contacts with persistent cough and their serum antibodies to pertussis toxin were positive. Sixteen infants with pertussis had the paroxysms of frequent and rapid coughs while another 5 with pertussis had long inspiratory effort accompanied by a high-pitched "whoop" at the end of paroxysms. Seven infants with pertussis had conjunctiva bleeding, a special sign of pertussis. Ten infants had lymphocytosis with a predominant elevation of lymphocytes. CONCLUSIONS: B. pertussis is an important pathogen for the infants under 3 months with persistent cough. Multiplex PCR may be used to identify B. pertussis with a high sensitivity. The unrecognized close family members of the infants with pertussis are probably an important source of infection.

[Expression of nuclear factor-kappaB and its inhibitor in alveolar macrophages of patients with neonatal hyaline membrane disease].

OBJECTIVE: Inflammatory reaction and injury in immature lungs are associated with activation of nuclear factor-kappa B (NF-kappaB) to trigger proinflammatory cytokine release, but the mechanism thereof is not fully understood. The present study was conducted to understand possible relationship between expression of NF-kappaB and its inhibitor and severity and outcome of neonates with hyaline membrane disease (HMD). METHODS: Serial samples of bronchoalveolar lavage fluid (BALF) were obtained during mechanical ventilation from 31 preterm infants with HMD. These infants were divided into two groups: survivors group [n = 22, birth weight (1500 +/- 320) g and gestational age (31.2 +/- 1.8) weeks] and nonsurvivors group [birth weight (1340 +/- 280) g, gestational age (30.8 +/- 2.1) weeks]. Nineteen preterm infants [birth weight (1470 +/- 280) g, gestational age (30.6 +/- 1.9) weeks] without respiratory disorders were enrolled as control subjects. Alveolar macrophages (AM) were isolated by differential adherence. AM was cultured and treated with lipopolysaccharide (LPS) for 1 hr. Then, nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for NF-kappaB expression. NF-kappaB inhibitor (IkappaB-alpha protein) in cytoplasmic extracts was detected by using Western blotting and IL-1beta and IL-8 in BALF by enzyme-linked immunosorbent assay (ELISA). RESULTS: NF-kappaB complexes were observed by EMSA, they were characterized by competition with cold oligonucleotide and p65-specific antibodies. The addition of an excess of cold oligonucleotide, corresponding to the NF-kappaB binding site, turned off the signal of the band, showing that the band was specific. An excess of an irrelevant oligonucleotide (corresponding to the SP-1) did not show any effect. The addition of an anti-p65 antibody caused the supershift of the two upper bands. After EMSA, the NF-kappaB complexes were quantified by using a ImageQuant software. NF-kappaB expression in AM at 24 hrs was higher in all the patients with HMD as compared with control subjects (survives/control, 34.1 vs 11.4 RDU, P < 0.01; nonsurvivors/control, 55.2 vs 11.4 RDU, P < 0.01). The NF-kappaB expression in AM at 72 hrs was higher than that in control subjects but not for nonsurvivors (survivors/control, 47.8 vs 25.6 RDU, P < 0.01; nonsurvivors/control, 21.8 vs 25.6, P > 0.05). The NF-kappaB expression in AM from nonsurvivors was depressed at 72 hrs as compared to 24 hrs (21.8 vs 55.2, P < 0.01), whereas the NF-kappaB expression in AM from survivors was still higher at 72 hrs than that at 24 hrs (47.8 vs 34.1, t = 4.43, P < 0.01). CONCLUSION: Altered NF-kappaB activation in AM of BALF of neonates with HMD was observed, and it may be mediated by decreased IkappaB synthesis, increased IkappaB degradation, or both. In HMD nonsurvivors NF-kappaB translocation was hampered upon LPS activation.

Nuclear factor-kappa B expression in alveolar macrophages of mechanically ventilated neonates with respiratory distress syndrome.

BACKGROUND: Inflammatory reaction and injury in mature lungs are associated with activation of nuclear factor-kappaB (NF-kappaB) to trigger proinflammatory cytokine release. In preterm infants with immature lungs, this mechanism is not yet fully understood, therefore we investigated this mechanism in mechanically ventilated neonates with respiratory distress syndrome (RDS). METHODS: Serial samples of the airway aspirates (AA) were obtained during mechanical ventilation from 21 preterm infants with RDS, of which 12 were survivors (birth weight 1.48 +/- 0.32 kg and gestational age 31 +/- 1.5 weeks) and 9 nonsurvivors (1.34 +/- 0.31 kg and 30 +/- 2 weeks). Seven neonates matched for age and birth weight without respiratory disorders served as controls. Alveolar macrophages (AM) of AA were isolated by differential adherence, some were cultured with lipopolysaccharide (LPS) for 1 h. Then, nuclear extracts of AM were analyzed by electrophoretic mobility shift assay for NF-kappaB expression. The NF-kappaB inhibitor (IkappaB-alpha protein) in cytoplasmic extracts was detected by Western blot, and concentrations of IL-1beta and IL-8 in AA by enzyme-linked immunosorbent assay (ELISA). RESULTS: On day 2 NF-kappaB expression in AM was significantly increased in the survivors and nonsurvivors at 33.3 +/- 9 and 54.8 +/- 10.2 relative density units (RDU) compared to control infants (11.1 +/- 6.7; p < 0.01). Expression of IkappaB-alpha was significantly higher in controls than that in the survivors and nonsurvivors on days 2 and 4. Moreover, in the nonsurvivors of RDS, expression of NF-kappaB was decreased following LPS stimulation in vitro on day 4. IL-1beta and IL-8 levels in the AA supernatant were higher in the survivors than in controls on days 2 and 4, but lower than those of the nonsurvivors on day 2. There were close correlations between the expression of NF-kappaB and levels of IL-1beta (r = 0.78, p < 0.01), and IL-8 (r = 0.81, p < 0.01) in AA. CONCLUSION: There were alterations in NF-kappaB activity in the AM of mechanically ventilated preterm neonates with RDS, mediated by decreased synthesis and increased degradation of IkappaB.