To explore the effect of advanced glycation end-products(AGEs)on cell viability and level of reactive oxygen species(ROS)in MIN6 cells. After intervention of various concentrations(100,200, and 400 mg/L)of AGEs for some time, cell viability was detected by MTT assay. 2', 7'-dichlorofluorescein diacetate(DCFH-DA)was used as a reactive oxygen species capture agent. The fluorescent intensity of 2', 7'-dichlorofluorescein(DCF), which was the product of cellular oxidation of DCFH-DA, was detected by flow cytometry. The level of ROS and insulin secretion was thus measured. Viability of MIN6 cells was inhibited by AGEs in a dose and time dependent manner(P＜0.05).Intracellular fluorescent intensity of DCF was markedly elevated in the AGEs groups as compared with that in the control group(P＜0.05).Insulin secretion was decreased in the AGEs groups than that in the control group(P＞0.05). The results suggest that AGEs inhibit the viability and induce oxidative stress in MIN6 cells by overproduction of ROS.

This study was aimed to improve the sensitivity of magnetic induction phase shift detection system for cerebral hemorrhage. In the study, a cerebral hemorrhage model with 13 rabbits was established by injection of autologous blood and the cerebral hemorrhage was detected by utilizing magnetic induction phase shift spectroscopy (MIPSS) detection method under the feature band. Sixty five groups of phase shift spectroscopy data were obtained. According to the characteristics of cerebral hemorrhage phase shift spectroscopy under the feature hand, an effective method, B-F distribution, to diagnose the severity of cerebral hemorrhage was designed. The results showed that using MIPSS detection method under feature band, the phase shift obviously growed with increase of injection volume of autologous blood, and the phase shift induced by a 3-mL injection reached -7.750 3 degrees ± 1.420 4 degrees. B-F distribution could effectively diagnose the severity of cerebral hemorrhage. It can be concluded that the sensitivity of the cerebral hemorrhage magnetic induction detection system is improved by one order of magnitude with the MIPSS detection method under the feature band.
Animals ; Cerebral Hemorrhage ; diagnosis ; Magnetic Phenomena ; Magnetics ; Rabbits ; Spectrum Analysis ; methods

Antibodies, Monoclonal ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; genetics ; metabolism ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; diagnosis ; genetics ; metabolism ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the role of autophagy in quercetin (Que)-induced apoptosis in human bladder carcinoma BIU-87 cells in vitro.

METHODSTo determine the proliferative inhibition by MTT colorimetric assay after treating BIU-87 cells with quercetin at various concentrations. To identify autophagy and apoptosis in the BIU-87 cells after Que treatment by monodansylcadaverin (MDC) and Hoechst 33258 fluorescent staining, respectively. To examine the cytotoxic effect of Que and influence of autophagy on apoptosis by studying LDH leakage rate and flow cytometry, after blocking the autophagy with 3-methlyadenine (3-MA), a specific autophagy inhibitor.

RESULTSThere was an obvious inhibitory effect of Que on the proliferation of BIU-87 cells in a time- and dose-dependent manner. The inhibition rate of BIU-87 cells after 200 µmol/L Que treatment for 72 hours was 89.2%. Autophagy and apoptosis were induced and detected in Que-treated BIU-87 cells and autophagy occurred earlier than apoptosis. The apoptosis peak became much higher after the autophagy was blocked. Whenever the autophagy was blocked before or after Que treatment, the Que-induced cytotoxicity in BIU-87 cells was enhanced.

CONCLUSIONSQuercetin significantly inhibits the proliferation of BIU-87 cells, and the autophagy is induced earlier than apoptosis. In the process of Que-induced apoptosis of BIU-87 cells, autophagy may play a protective role at the initiation phase, delay apoptosis and reduce the Que-induced death of BIU-87 cells.

Adenine ; analogs & derivatives ; pharmacology ; Antioxidants ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Autophagy ; drug effects ; physiology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; L-Lactate Dehydrogenase ; drug effects ; metabolism ; Quercetin ; administration & dosage ; pharmacology ; Urinary Bladder Neoplasms ; pathology

Objective:To understand the effects of proton and heavy ion radiotherapy on nutritional status in patients with malignant tumors and to analyze the influencing factors of adverse events.Methods:Patients with malignant tumors who received proton and heavy ion therapy between October 2016 and September 2021were retrospectively included. The demographic characteristics, clinical diagnosis, radiotherapy regimen, nutritional indicators and adverse events were collected. Paired t test was used to analyze the changes in nutritional status before and after treatment and logistic regression was used to analyze the influencing factors of adverse events. Results:A total of 2,390 patients were enrolled and were stratified into 4 groups according to different radiotherapy regimen, namely proton, heavy ion, proton + heavy ion and photon + heavy ion radiotherapies. The prevalence of nutritional risk were 17.5% and 27.8% at admission and discharge, respectively. The prevalence of nutritional risk at discharge were 73.9% ( χ2 = 237.149, P < 0.01) in patients who received photon + heavy ion radiotherapy and 30.8% ( χ2 = 36.925, P < 0.01) in those who received proton + heavy ion radiotherapy. The prevalence of critical weight loss at discharge was 14.1%, with the absolute weight loss of 4.84 kg ( t = 11.716, P < 0.01) and 1.52 kg ( t = 29.530, P < 0.01) in photon + heavy ion radiotherapy and proton + heavy ion radiotherapy groups, respectively. All groups showed significant changes in serum albumin (ALB) and total lymphocyte count (TLC). Specifically, photon + heavy ion and proton + heavy ion therapy had a greater effect on serum ALB and TLC, with a decrease of 2.88 g/L and 2.18 g/L for ALB as well as a decrease of (1.06×10 9) /L and (0.80×10 9) /L for TLC ( P < 0.01). Multivariate logistic regression analysis showed that nutritional risk at admission and concurrent chemotherapy were independent factors for adverse events of proton and heavy ion radiotherapy ( OR = 1.404, 95% CI: 1.039 to 1.898; OR = 2.370, 95% CI: 1.781 to 3.154). Compared with heavy ion radiotherapy, the other 3 groups had more adverse events (proton, OR = 3.982, 95% CI: 2.533 to 6.259; proton + heavy ion, OR = 4.995, 95% CI: 3.688 to 6.766; photon + heavy ion, OR = 7.716, 95% CI: 5.079 to 11.720). Conclusions:Patients receiving proton and heavy ion therapy showed poorer nutritional status. Photon + heavy ion therapy had the greatest impact on nutritional status. Nutritional risk at admission and concurrent chemotherapy were independent factors for adverse events in patients receiving proton and heavy ion therapy.

Delphi Technique ; Humans ; Radiculopathy ; drug therapy ; surgery ; therapy

OBJECTIVETo evaluate the diagnostic accuracy of 320-slice CT coronary angiography (CTA) in the evaluation of in-stent restenosis (ISR, ≥50% luminal narrowing) in comparison with quantitative coronary angiography (CAG).

METHODSA total of 69 patients with previous stent implantation who underwent both CTA and CAG were prospectively included. We assessed diagnostic valve for ISR with CTA in comparison with CAG.

RESULTSA total of 110 stents were implanted in these patients.CAG identified 14 ISR. CTA correctly identified 13 ISR and misdiagnosed 5 ISR in stents without ISR. Besides, 6 stents could not be evaluated by CTA due to unsatisfied image quality. Accordingly, sensitivity, specificity, positive and negative predictive value of CTA for diagnosing ISR were 93%, 89%, 54% and 99%, respectively. The image quality of CTA was significantly better in larger stents (percentages of good and moderate stent image of ≥3.0 mm and <3.0 mm: 56% vs. 27%, 25% vs. 49%) and which was linked with better diagnostic coincidence rate (95% vs. 78%) for larger stents. The image quality of CTA was significantly better in stents with thinner stent strut thickness (percentages of poor CTA stent image quality of stent strut thickness<140 µm and ≥140 µm: 12% vs. 45%, P<0.01) and which was associated with better diagnostic coincidence rate for stents with thinner stent strut thickness (94% vs. 76%, P<0.05). The image quality of CTA was also significantly better in single stent (percentages of poor CTA stent image quality of single stent vs. overlap and dedicated stent: 17% vs. 36%, P<0.05). However, heart rate (≥65 beats/min vs. <65 beats/min) during CTA acquisition was not associated with image quality and the diagnostic coincidence rate (all P>0.05).

CONCLUSIONSOur results indicate that 320-slice CTA allows accurate noninvasive assessment of significant in-stent restenosis in selected patients. Stents with a large diameter and thin struts are associated with better image quality and higher diagnostic accuracy.

Aged ; Aged, 80 and over ; Coronary Angiography ; Coronary Restenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Predictive Value of Tests ; Prospective Studies ; Sensitivity and Specificity ; Stents ; Tomography, X-Ray Computed ; methods

OBJECTIVETo develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria.

METHODSType-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products.

RESULTSTwenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification.

CONCLUSIONThe novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.

Bacteria ; classification ; genetics ; isolation & purification ; DNA, Bacterial ; analysis ; Food Contamination ; analysis ; Food Microbiology ; methods ; Foodborne Diseases ; microbiology ; Humans ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo explore the synergetic effect of norcantharidin (NCTD) and adriamycin (ADR) on the proliferation and apoptosis of multiple myeloma (MM) cells.

METHODSHuman MM cell line U266 cells were treated with NCTD alone (10 µmol/L) or in combination with ADR (0.25 µmol/L). MTT and Annexin V/PI staining were used to determine cell viability and apoptosis. The protein expression of nuclear factor-κB P65 (NF-κB P65), phosphorylated NF-κB p65 (p-NF-κB p65), NF-κB P65 inhibitor IκBα, phosphorylated IκBα (p-IκBα), survivin, Bcl-2 and Bax were determined by Western blot. Immunohistochemistry was used to determine the expression of vascular endothelial growth factor (VEGF).

RESULTS(1) NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by ADR. The combination of NCTD and ADR had synergistic anti-proliferation effect. (2) Combination of ADR and NCTD downregulated the expression of nuclear NF-κB P65 and cytoplasm p-IκBα induced by ADR. The expression of nuclear NF-κB P65 and cytoplasm p-IκBα decreased from 2.08 ± 0.29 and 0.39 ± 0.07 to 0.48 ± 0.08 and 0.02 ± 0.01 respectively, while the expression of the cytoplasm NF-κB P65 and IκBα were unchanged in the ADR alone group and the combined group. (3) The expression of survivin and bcl-2 decreased from 0.31 ± 0.05 and 0.23 ± 0.05 to 0.03 ± 0.02 and 0.05 ± 0.02, while the expression of Bax increased from 0.46 ± 0.06 to 0.62 ± 0.08 respectively in ADR alone group and combined group. (4) The positive rate of VEGF in ADR group and combination group were (44.6 ± 4.4)% and (27.0 ± 2.1)% respectively, indicating that NCTD could potentiate the inhibition effect on VEGF induced by ADR.

CONCLUSIONSThe results suggest that NCTD can potentialize the chemosensitivity of multiple myeloma cells to ADR through regulating NF-κB/IκBα signaling pathway and NF-κB-regulated gene products including survivin, Bcl-2, Bax and VEGF.

Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Doxorubicin ; pharmacology ; Humans ; I-kappa B Proteins ; metabolism ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; metabolism ; NF-KappaB Inhibitor alpha ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction ; drug effects ; Transcription Factor RelA ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the development and prognosis of the acute flaccid paralysis (AFP) associated with enterovirus 71 (EV71) infection through clinical follow-up study for clinical and magnetic resonance imaging (MRI) features based on the research progress of virology and pathology.

METHODSixteen children with HFMD associated with AFP in hospital from May 1, 2011 to August 31, 2011 were investigated and the patients received intensive rehabilitation training. The 16 cases were divided into two groups (the recovery or the sequela) by if the muscle strength recovered to level 4 after intensive rehabilitation. The MRI findings of 15 children were analyzed and among them, 6 patients were reexamined after one month. The clinical markers were compared between groups including course of disease, WBC, WBC in cerebrospinal fluid (CSF), ventilator support, therapy, the worst muscle strength, the initial tendon reflex, the muscle atrophy, and multi-limb paralysis. The data were analyzed by t test and χ2 test with SPSS10.0.

RESULTAll the 16 children were infected with enterovirus 71 (EV71). The myodynamia of 7 children were level 0, 4 children had serious upper limbs paralysis. The neck muscle in 3 cases and the brain stem motor ruckus in 4 cases were involved. The ankle clonus of non-completely paralyzed limbs in 14 cases occurred during rehabilitation. Eight children had the better prognosis, the other 8 children had sequela. 0 level muscle strength (0 case vs. 7 cases, χ2=12.4), the initial tendon reflex (2 cases vs. 8 cases, χ2=9.6), obvious muscle atrophy (0 case vs. 8 cases, χ2=16), were significantly different in the children with the recovery when compared to the sequela (P<0.01). The severe upper limbs paralysis had the worse prognosis than the severe lower limbs paralysis. MR imaging showed signs of spinal nerve root inflammation and the bilateral hyperintense lesions, symmetrical in the posterior portions of the medulla, pons, and asymmetrical in the ventral horns of cervical spinal cord. Signal enhancement occurred only in the early MRI examination.

CONCLUSIONIn the evolution of AFP due to EV71 infection, the upper motor neuron damage is common, the prognosis is related with the severity of early paralysis and neuron damage. MR imaging is helpful to understand the pathological mechanism of AFP.

Child, Preschool ; Enterovirus A, Human ; pathogenicity ; Female ; Follow-Up Studies ; Hand, Foot and Mouth Disease ; diagnosis ; pathology ; virology ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Paralysis ; diagnosis ; pathology ; virology ; Prognosis