We have found the parallel relationship between the live cell number of several tumor cell lines and the formation of MTT formazan with MTT colorimetric assay. The MTT colorimetric assay was compared with ~3H-Tdr incorporation assay for the proliferation of mouse spleen cells induced by mitogens or the activity of rat/mouse IL2,the results suggested that the ~3H-Tdr incorporation assay can be replaced by MTT colorimetric assay. MTT colorimetric assay have the advantages of simplicity, rapidity,save any radioisotope and no specific equivepment, etc, so under some conditions it would be a useful method for measuring cell proliferation or cytotoxicity in a laboratory.

OBJECTIVE: To observe the chemical groups changing in rat kidney with regard to fatal hyperthermia by Fourier transform infrared microspectroscopy (FTIR-MSP) and to provide a new method to diagnose fatal hyperthermia. METHODS: Rats were sacrificed by hyperthermia, brainstem injury, massive hemorrhage and asphyxiation and divided into groups. The renal samples were dissected immediately after death. The data of infrared spectroscopy in glomerulus were measured by FTIR-MSP. RESULTS: The absorbances of 3290, 3070, 2850, 1540 and 1396 cm(-1) significantly increased (P < 0.05), and the ratios of Al650/A3290 and A1650/A1540 significantly decreased (P < 0.05) in group of hyperthermia. CONCLUSION FTIR-MSP can analyze the changes of chemical groups of kidney as an auxiliary diagnosis for discriminating hyperthermia with other causes of death.
Animals ; Fever/mortality* ; Fourier Analysis ; Kidney/metabolism* ; Microspectrophotometry ; Rats ; Spectroscopy, Fourier Transform Infrared/methods*

Objective To investigate the effect of 9-cis retinoid acid(c-RA),a retinoid X receptor(RXR)agonist,on hydrogen peroxide(H2O2)induced apoptosis in cultured rat neonatal cardiomyoeytes,and to explore the mechanism.Method Cultured cardiomyocpes were randomly divided into three groups:normal group treated with vehicle(N group),H2O2 group treated with 100 μmol/L H2O2(H group),and c-RA group pretreated with 100nmol/L c-RA(H+R group).Cell viability was detected by MTT.Morphological changes of apoptotic cardiomyocytes were observed by Hoechst 33258 staining under fluorescence microscope.The apoptotic rate was determined by flow cytometry.Mitochondrial membrane potential(△(ψ)m)was measured by JC-1 dye.Cellular reactive oxygen species(ROS)production was detected by CM-H2DCFDA fluorescent probe.All measurement data wIe expressed as(x±s),and statistically analyzed using one-way ANOVA analysis and Dunnett test.Differences were considered significant when P was＜0.05.Results Treatment with c-RA significantly enhanced cell viability,reduced apoptosis ratio,stabled mitoehondrial membrane potential and reduced level of cellular reactive oxygen species.Conclusions RXR agonist c-RA inhibits H2O2-induced myocyte apoptosis in cultured rat neonatal cardiomyocytes,which may be related to alleviate oxidative stress injury.

OBJECTIVETo observe the relationship between the indicators of lung injury and the ratio of angiopoietin-2 (ang-2) to angiopoietin-1 (ang-1) in serum of rats with acute lung injury induced by phosgene.

METHODSEighty four adult male SD rats were randomly divided into 13 phosgene groups and 1 control group. Control group was exposed to fresh air for 5 minutes; the phosgene groups were exposed to phosgene (100% purity of 8.33 mg/L phosgene ) for 5 minutes. At different time points after exposure, the wet/dry weight ratio of mouse lungs was examined, the levels of ang-1 and ang-2 in serum of rats were detected with ELISA assay, and the white blood cell count and total protein levels in bronchoalveolar lavage fluid (BALF) of rats were measured.

RESULTSIn 48 h after exposure,the serum ang-2, lung wet/dry weight ratio, BALF white blood cell count, protein concentration tended to increase with time, and the serum ang-1 tended to decrease with time. The ratios of ang-2 to ang-1 at 6, 8, 10, 12, 14, 16, 18, 20, 22, 24 and 48 h after exposure in phosgene groups were significantly higher than that in control group (P < 0.05 or P < 0.01). There were the positive correlations between ang-2 and lung wet/dry ratio, BALF white blood cell count or protein concentration (P < 0.01). There were the negative correlations between ang-1 and lung wet/dry ratio or BALF white blood cell count (P < 0.01). There were the positive correlations between the ratio of ang-2 to ang-1 and lung wet/dry ratio, BALF white blood cell count or protein concentration (P < 0.01).

CONCLUSIONAfter exposure of rats to phosgene, there is a positive correlation between the ratio of ang-2 to ang-1 and lung injury indicators, which may be an early indicator of the severity of phosgene-induced acute lung injury.

Acute Lung Injury ; chemically induced ; metabolism ; Angiopoietin-1 ; metabolism ; Angiopoietin-2 ; metabolism ; Animals ; Disease Models, Animal ; Male ; Phosgene ; toxicity ; Rats ; Rats, Sprague-Dawley

Objective To investigate the number and activities changes of peripheral blood endothelial progenitor cells (EPCs) in continuous ambulatory peritoneal dialysis (CAPD) patients,and explore the connection between EPCs' number and the levels of advanced glycosylation end products (AGEs),homocysteine (Hcy) and C-reactive protein (CRP).Methods Twenty-five CAPD patients and thirty healthy volunteers were involved.Total mononuclear cells (MNCs) were isolated from peripheral blood of patients.EPCs were characterized as adherent cells by double staining of FITCUEA-1 and DiL-AcLDL binding,and were further demonstrated by positive cells of CD34,CD133 and KDR using flow cytometry.The abilities of cell proliferation,adhesion and migration were further observed by fluorescent microscope.The correlations between the CEPCs' number and the levels of AGEs,Hcy and CRP were analyzed.Results The number and activities including migration and adhesion of EPCs in CAPD group were significantly lower than control group (P ＜ 0.05).The levels of serum AGEs,Hey and CRP in CAPD patients were increased (all P＜0.05) and had negative correlation with EPCs' number.Conclusions The number and activities of EPCs decrease in patients with CAPD,and EPCs' number is negatively correlated to the levels of AGEs,Hcy and CRP.

Objective:To investigate the acceptance of advance care planning and its influencing factors in heart failure patients.Methods:A total of 208 patients with heart failure were surveyed by general data questionnaires and advance care planning acceptance questionnaires.Results:The total score of advance care planning acceptance of heart failure patients was (44.26 ± 11.73), the score of feeling dimension was (13.67 ± 5.72), the score of attitude dimension was (30.59 ± 6.33). 53.4%(111/208) of patients were willing to accept the talking about advance care planning. Regression analysis results showed that education level, New York Heart Association (NYHA) classification, communication status with medical staff and whether they had received life-sustaining treatment were important factors influencing of the acceptance of advance care planning in patients with heart failure.Conclusion:Patients with heart failure had higher acceptance of advance care planning. In clinical work, it is necessary to strengthen the scientific popularization of advance care planning in patients with low education level, low NYHA grade and no exposure to life-sustaining treatment. And strengthen the daily communication with patients to prepare for the follow-up advance care planning related communication.

Objective To explore the effect of action verb processing on the excitability of the motor cortex and any effect on the upper limb motor function of stroke survivors.Methods Experiment 1:The motor evoked potential by transcranial magnetic stimulation (TMS-MEP) was measured while 18 healthy subjects were processing concrete action verbs or abstract verbs.Experiment 2:Eight hemiplegic stroke survivors were asked to read silently text describing concrete hand actions,and then repeat and explain the meaning of the action verbs used in the text.This was repeated for 30 minutes per day,5 days per week for 3 weeks.In the 2nd week the same training was conducted except that the text was about abstract verbs.The modified Ashworth scale (MAS),Fugl-Meyer assessment (FMA),the Hong Kong version of the functional test for a hemiplegic upper extremity (FTHUE-HK) and a simple test for evaluating hand function (STEF) were used to assess upper limb motor function before and after the training each week.Results Experiment 1:Processing the concrete action verbs induced significantly greater MEP than processing the abstract verbs.Experiment 2:The average FMA and STEF ratings improved significantly after the training each week.Training with the concrete action verbs resulted in significantly better FMA and STEF scores than with the abstract verbs.The average MAS score increased significantly week by week.There was significantly more improvement in the average FTHUE-HK rating after the 1st and 3rd week of training than after the 2nd week.Conclusion Action verb processing induces greater motor cortex excitation than abstract verb processing among healthy subjects and better improves the upper limb motor function of stroke survivors.

Objective To investigate the mRNA and protein expression levels of interferon(IFN)-?in the peripheral blood of patients with systemic lupus erythematosus(SLE),to analyze the relationship between IFN-?and disease activity,and to evaluate the role of IFN-?in the pathogenesis of lupus.Methods SYBR green dyeⅠbased real-time quantatives PCR method was used to compare the mRNA expression levels of IFN-?in the peripheral blood leucocyte of SLE patients and healthy controls.Surum levels of IFN-?were measured with ELISA method.Results IFNA1 mRNA expression level in SLE patients(2.8?3.5)was signifi- cantly lower than that of normal controls(12.7?10.7,P=0.000).There was no significant difference between patients treated with glucocorticoid and those without in the expression level of IFNA1(P=0.549).Serum levels of IFN-?in SLE patients was significantly higher than that of normal controls(P=0.003).The SLEDAI score and anti-dsDNA antibody correlated positively,and complement components C3,C4 and leukocytes correlated negatively with serum concentration of IFN-?.IFN-?level correlated with the presence of fever and rash. Conclusion The close relationship between IFN-?serum level and disease activity in SLE patients suggests that IFN-?might be of importance in the disease process.

BACKGROUNDLiver targeting drug delivery systems can improve the curative effects and relieve the cytotoxicity of the chemotherapy drugs in the treatment of liver diseases. Nanoparticles carrying therapeutic drugs are currently under hot investigation with great clinical significance. This study was aimed to investigate the different tissue distribution of the adriamycin polybutylcyanoacrylate nanoparticle (ADM-PBCA-NP) in the mice body after an injection via lateral tail vein, and to study the liver targeting effects of ADM-PBCA-NP in different diameters on normal mice liver.

METHODSOne hundred and eighty Kunming mice were randomly divided into 6 groups with 30 mice in each group (5 treatment groups of ADM-PBCA-NP in the different diameter ranges, non-conjugated free adriamycin injection was employed as the control group). A single dose of either conjugated or free adriamycin equaled 2 mg/kg of body weight was delivered via the tail vein. Five mice in each trail were sacrificed at 5, 15, 30 minutes, 1, 5 and 12 hours postinjection, respectively. The adriamycin concentrations in the respectively collected liver, kidney, spleen, heart, lung and plasma were demonstrated using a high performance liquid chromatography with fluorescence detector.

RESULTSCompared with the control group, adriamycin was hardly detected in the heart muscle of the treatment groups (P < 0.05). The nanoparticle-conjugated adriamycin was cleaned up quickly from the kidney tissue. The adriamycin concentrations of the mice liver and spleen in the experimental groups were significantly higher than that in the control group, except for the group with the nanoparticles diameters of (22.3 +/- 6.2) nm (P < 0.05). The ADM-PBCA-NP in (101.0 +/- 20.3) nm diameter had the highest liver distribution, and the second highest adriamycin distribution in liver was the group of (143.0 +/- 23.5) nm diameter (P < 0.05). Moreover, adriamycin was released slowly in the liver during the detection period in the experimental groups. ADM-PBCA-NP in (22.3 +/- 6.2) nm diameter was not distributed in the tissue of the liver, kidney, heart, spleen, and lung.

CONCLUSIONSADM-PBCA-NP in 100 - 150 nm diameter range has the best liver targeting with a characteristic of slow medicine release. It also decreases the medicine distribution in the heart, kidney and lung. In the treatment of liver cancer, the polybutylcyanoacrylate nanoparticles system has a good liver targeting ability, which increases the anticancer activity and markedly decreases the toxicity of adriamycin.

Animals ; Antineoplastic Agents ; administration & dosage ; Doxorubicin ; administration & dosage ; Drug Delivery Systems ; Enbucrilate ; administration & dosage ; Liver ; metabolism ; Mice ; Nanostructures ; Tissue Distribution

Objective: To investigate the risk factors of ventricular arrhythmias in patients with Brugada syndrome. Methods: Clinical data of 60 Brugada syndrome patients admitted in the department of cardiology of the First Affiliated Hospital of Nanjing Medical University from March 2003 to December 2016 were collected and retrospectively analyzed. The age at diagnosis was (43.2±13.1) years (0.6-83.0 years), 98.3% were males (n=59), and the patients were followed up to (92±41) months (12-169 months). The 12-lead surface electrocardiogram (ECG) recorded at the time of diagnosis and showing the highest type 1 ST elevation, either spontaneously or after provocative drug test, was used for the analysis. Patients were divided into ventricular arrhythmia (VA, n=12) group and non-ventricular arrhythmia (non-VA, n=48) group depending on the presence or absence of clinical VA event. The demographic data and ECG data of the 2 groups were compared, and the independent risk factors of VA events were analyzed by stepwise logistic regression. Results: Incidence of family history of sudden death (7/12 vs. 22.9% (11/48)) and percentage of type 1 ST elevation in the peripheral ECG leads (6/12 vs. 16.67% (8/48)) were significantly higher in VA group than in non-VA group (both P<0.05). Max Tpeak-Tend (Max-Tpe) interval ((144±53)ms vs. (110±16)ms) and dispersion of Tpe ((74±50)ms vs. (43±17)ms) were significantly higher in VA group than in non-VA group (both P<0.05). The area under receiver operating characteristic (ROC) curves for the Max-Tpe interval was 0.693 and Max-Tpe interval ≥140 ms was determined as an optimized cutoff point with increased risk of VA event, which had a sensitivity of 50.0%, a specificity of 98.0%, a positive predictive value of 85.7%, and a negative predictive value of 88.7% for predicting VA event. The ROC curves for the dispersion of Tpe was 0.775 and dispersion of Tpe ≥45 ms was determined as an optimized cutoff point for predicting VA event, which had a sensitivity of 91.7%, a specificity of 64.6%, a positive predictive value of 39.3%, and a negative predictive value of 96.9% for predicting VA event. In multivariate analysis, Max-Tpe interval ≥140 ms (OR=27.53, 95%CI 1.07-706.77, P=0.045) and family history of sudden death (OR=24.63, 95%CI 2.05-295.38, P=0.011) were found to be the independent risk factors of arrhythmic events. Conclusions Max-Tpe interval ≥140 ms and family history of sudden death are risk factors of VA event in included patients with Brugada syndrome.