Group EI Research Articles

Cardiopulmonary exercise testing in long covid shows the presence of dysautonomia or chronotropic incompetence independent of subjective exercise intolerance and fatigue

BackgroundAfter COVID-19 infection, 10–20% of patients suffer from varying symptoms lasting more than 12 weeks (Long COVID, LC). Exercise intolerance and fatigue are common in LC. The aim was to measure the maximal exercise capacity of the LC patients with these symptoms and to analyze whether this capacity was related to heart rate (HR) responses at rest and during exercise and recovery, to find out possible sympathetic overactivity, dysautonomia or chronotropic incompetence.MethodsCardiopulmonary exercise test was conducted on 101 LC patients, who were admitted to exercise testing. The majority of them (86%) had been treated at home during their acute COVID-19 infection. Peak oxygen uptake (VO2peak), maximal power during the last 4 min of exercise (Wlast4), HRs, and other exercise test variables were compared between those with or without subjective exercise intolerance, fatigue, or both.ResultsThe measurements were performed in mean 12.7 months (SD 5.75) after COVID-19 infection in patients with exercise intolerance (group EI, 19 patients), fatigue (group F, 31 patients), their combination (group EI + F, 37 patients), or neither (group N, 14 patients). Exercise capacity was, in the mean, normal in all symptom groups and did not significantly differ among them. HRs were higher in group EI + F than in group N at maximum exercise (169/min vs. 158/min, p = 0.034) and 10 min after exercise (104/min vs. 87/min, p = 0.028). Independent of symptoms, 12 patients filled the criteria of dysautonomia associated with slightly decreased Wlast4 (73% vs. 91% of sex, age, height, and weight-based reference values p = 0.017) and 13 filled the criteria of chronotropic incompetence with the lowest Wlast4 (63% vs. 93%, p < 0.001), VO2peak (70% vs. 94%, p < 0.001), the lowest increase of systolic blood pressure (50 mmHg vs. 67 mmHg, p = 0.001), and the greatest prevalence of slight ECG-findings (p = 0.017) compared to patients without these features. The highest prevalence of chronotropic incompetence was seen in the group N (p = 0.022).ConclusionsThis study on LC patients with different symptoms showed that cardiopulmonary exercise capacity was in mean normal, with increased sympathetic activity in most patients. However, we identified subgroups with dysautonomia or chronotropic incompetence with a lowered exercise capacity as measured by Wlast4 or VO2peak. Subjective exercise intolerance and fatigue poorly foresaw the level of exercise capacity. The results could be used to plan the rehabilitation from LC and for selection of the patients suitable for it.

Differences in extracellular fluid volume between acute heart failure patients with and without high systolic blood pressure.

Some reports have suggested that hypertensive acute heart failure (AHF) is caused by intravascular congestion, not interstitial congestion. We evaluated the differences in extracellular fluid volume assessed by bioelectrical impedance analysis (BIA) between AHF patients with and without high systolic blood pressure (sBP). This prospective single-centre study (UMIN000030266) included 178 patients hospitalized due to AHF between September 2017 and August 2018. We calculated extracellular water (ECW), intracellular water (ICW), total body water (TBW), and ECW-to-TBW ratio (oedema index: EI) by BIA and evaluated conventional parameters as follows: weight, N-terminal pro brain natriuretic peptide values, and echocardiography parameters on admission and before discharge. One-year outcomes included all-cause death and re-admission due to heart failure. We compared patients with sBP>140mmHg on admission [clinical scenario 1 (CS1) group] and with sBP of ≤140mmHg on admission (non-CS1 group). The mean age of the patients was 79.5±11.1years, and 48.9% of the patients were female. EI on admission of 83 patients in the CS1 group was lower than that of 95 patients in the non-CS1 group. The change in EI from admission to before discharge was no significant in the CS1 group but was significant in the non-CS1 group. Comparing the changes from admission to before discharge between the CS1 and the non-CS1 group, delta ECW, delta ICW, delta TBW, and delta EI of the CS1 group were significantly smaller than those of the non-CS1 group. During the 1-year follow-up period after discharge of the 178 patients, the numbers of deaths and re-admissions due to acute HF were 26 (15%) and 49 (28%), respectively. Patients with high EI before discharge [>0.408 (median)] had significantly more cardiac events than patients with low EI [hazard ratio (HR): 2.15, 95% confidence interval (CI): 1.30-3.55]. Cox regression analysis revealed that higher EI as a continuous variable was significantly associated with worse outcome in non-CS1 group (HR: 1.46, 95% CI: 1.13-1.87), but not significantly associated with worse outcome in CS1 group (HR: 1.29, 95% CI: 0.98-1.69). EI on admission in patients with high sBP was not elevated, and changes in ECW, ICW, TBW, and EI in patients with high sBP were smaller than those in patients without high sBP. EI measured by BIA could distinguish AHF with interstitial or intravascular congestion.

The Efficacy of a Long-Acting Injectable Selenium Preparation Administered to Pregnant Ewes and Lambs.

Simple SummaryLack of selenium (Se) is a global problem that leads to increased exposure of animals to various diseases, as well as a reduction in productive and reproductive performance. Mineral supplementation can improve the immune system and could therefore be of assistance for lambs. We studied the effects of injectable supplementation with selenium. Supplementation of the Se lead to improved immune status, blood parameters, body weights, and average daily gain of lambs.The aim of this study was to evaluate the effects of a long-acting selenium (Se) preparation administered to sheep. The experiment was conducted on 30 dams and 36 lambs divided into three equal groups of 10 dams and 12 lambs each: Control—C, and two experimental groups—E (Se administered to pregnant ewes) and EI (Se administered directly to lambs after the colostral period). The Se preparation (Barium Selenate Injection, BVP Animal Care, 50 mg/mL) was administered by injection at 1 mL/50 kg (1 mg Se/kg) body weight (BW) to group E ewes in the third month of pregnancy (between 70 and 90 days) and to group EI lambs between 4 and 7 days of age. The following parameters were determined: Se concentration in the blood of ewes, milk yield, milk composition, Se concentration in milk; hematological, biochemical, and immunological parameters and Se concentration in the blood of lambs; growth rate and in vivo measurements of lean meat and fat content in lambs. Barium selenate significantly improved the Se status of dams and lambs, regardless of whether it was administered to pregnant ewes or directly to lambs in the first week of their life. The milk of ewes receiving the Se preparation was characterized by higher concentrations of fat and dry matter. The Se preparation induced significant changes in immunological parameters, thus enhancing defense mechanisms in lambs. The Se preparation exerted more stimulatory effects on humoral and cellular immune responses when administered directly to lambs after the colostral period (group EI) than to pregnant ewes (group E). The results of this study indicate that the long-acting Se preparation delivers benefits to sheep by boosting their immunity and, therefore, improving performance.

Systemic Effects of Photobiomodulation on the Morphology of the Thyroid and Sublingual Glands: A Study in Rabbits.

Objective: To investigate whether photobiomodulation (PBM) applied in a clinical situation with the purpose of improving the healing process of implants placed in the rabbit mandible would cause any morphological change in the thyroid and sublingual glands as a systemic effect of laser irradiation. Methods: Thirty-two New Zealand rabbits were randomly divided into four groups of eight animals each, one control group (CI, nonirradiated animals) and three experimental groups (EI, EII, and EIII) that received PBM postoperatively with an aluminum/gallium/arsenide laser diode (Theralase®) at a wavelength of 830 nm (infrared) and 50 mW output power applied to two irradiation fields per session, for a total of seven sessions. All rabbits underwent surgical extraction of the mandibular left incisor, followed by immediate placement of an osseointegrated implant in the fresh socket. The experimental groups EI, EII, and EIII received PBM at an energy density of 5, 2.5, and 10 J/cm2, respectively, per irradiation field. Results: There was no histomorphometric change in any of the groups. Conclusions: PBM, based on the irradiation protocol used in this study, does not cause morphological changes in the thyroid and sublingual glands when used to stimulate peri-implant bone healing in the rabbit mandible.

The Effect of Different Doses of Zearalenone in Feed on the Bioavailability of Zearalenone and Alpha-Zearalenol, and the Concentrations of Estradiol and Testosterone in the Peripheral Blood of Pre-Pubertal Gilts

The objective of this study was to determine the effect of long-term (48 days), per os administration of specific zearalenone (ZEN) doses (20 and 40 μg ZEN/kg BW in experimental groups EI and EII, which were equivalent to 200% and 400% of the upper range limit of the no-observed-adverse-effect-level (NOAEL), respectively) on the bioavailability of ZEN and the rate of changes in estradiol and testosterone concentrations in the peripheral blood of pre-pubertal gilts. ZEN and α-ZEL levels were similar until day 28. After day 28, α-ZEL concentrations increased significantly in group EI, whereas a significant rise in ZEN levels was noted in group EII. The presence of estradiol in peripheral blood plasma was not observed until day 20 of the experiment. Spontaneous secretion of estradiol was minimal, and it was determined at very low levels of up to 10 pg/mL in EI and EII groups. Testosterone concentrations ranged from 4 to 9 ng/mL in all groups. A decrease in the concentrations of both analyzed hormones was reported in the last stage of the experiment. The results of the experiment indicate that: (i) The bioavailability of ZEN in peripheral blood has low diagnostic value, (ii) exposure to low doses of ZEN induces minor changes in the concentrations of the analyzed hormones, which could lead to situational supraphysiological hormone levels and changes in endogenous hormonal balance.

The effect of estradiol on sevoflurane-induced epileptiform cortical EEG waves in neonatal rats

Objective To investigate the effect of estradiol on sevoflurane-induced epileptiform cortical EEG waves in neonatal rats. Methods Forty neonatal rats, aged 4-6 days, weighting 8-15 g, were divided into 5 groups according to random number table: normal saline control group (group C), estradiol group (group E), ICI182780 (group I), Formestane group (group O) and estradiol + ICI182780 (group EI). After establishing the monitoring model of cortical electroencephalogram in neonatal rats, EEG was recorded continuously for 30 minutes, then different treatment drugs were given according to different groups. After 30 minutes of continuous recording of EEG, anesthesia induction was started with inhalation of 6% sevoflurane for 3 minutes, followed by inhalation of 2.1% sevoflurane to maintain anesthesia. Sevoflurane anesthesia lasted for 1 hour.The righting reflex disappearance time was recorded after sevoflurane anesthesia.The total duration, single duration and frequency of EEG convulsion waves were recorded.Spike frequencies were recorded at different periods.Animal respiratory rate was recorded after sevoflurane induction. Results One rat in group I and one rat in group EI showed convulsion wave before treatment, which were excluded from the study.The righting reflex disappearance time of group S, group E, group I, group O and group EI were (24.4±2.5)s, (16.4±4.2)s, (31.8±5.2)s, (29.8±1.8)s, (24.8±2.7)s respectively, and there were statistically significant differences among the 5 groups (F=16.693, P 0.05). Compared with group S, the total duration, single duration and frequency of convulsion wave were significantly increased in group E during anesthesia, and the differences were statistically significant (P<0.05). Compared with group E and group S, the total duration, single duration and frequency of convulsion wave during anesthesia were significantly reduced in group I, group O and group EI, and the differences were statistically significant (P<0.05). Compared with group S, the spike frequency of group E increased significantly in each period, and the differences were statistically significant (P<0.05). The frequency of spike wave in group I, group O and group EI at 65-70 min and 90-95 min were lower than that in group S, and the differences were statistically significant (P<0.05). The frequency of spike wave between group O and group EI during 115-120 min was statistically different with group S and group E(P<0.05). Conclusion The mechanism of epileptiform EEG wave induced by sevoflurane anesthesia may be related with neuroactive steroidal estradiol. Key words: Sevoflurane; Neonatal rats; Epileptiform electroencephalography; Estradiol

OR-020 High-intensitive training leads to increasing apoptosis of podocyte in kidneys of rats

Objective Athletes often choose high-intensitive training load to improve athletic ability in their training cycle, its purpose is to pursue the super-compensation after training, but athletes and coaches frequently ignore the balance between load and reasonable recovery, which produce a sustained decline in athletic ability and physical function, and cause the viscera function disorder. High-intensitive training damages the structure of renal filtration barrier in rats, especially the destruction and fracture of Silt diaphragm ultrastructure, as well as the partial fusion of foot processes, etc., so that the large amount of macromolecular proteins in the blood leak out far beyond the threshold of renal tubular reabsorption and form exercise-induced proteinuria. Podocyte is one of the most important. This research establishes a model of rats which simulates the progressive-load training in the cycle of athletes. Observe the apoptosis of renal cells in rats by bioimaging. Determine whether long-term intensive training causes apoptosis in kidney of rats or podocytes, and detect the expression and distribution of Bcl-2 and Bax, and the protein expression of Caspase-3, which is the final regulatory factor of apoptosis pathway. Study the mechanism of the abnormal podocyte due to intensive training from the characterization of kidney to the molecular level, providing experimental basis for explaining the relationship between intensive-training and exercise-induced proteinuria to guide of scientific training. Methods This study selects 36 Sprague-Dawley rats, which are randomly divided into 3 groups: a control group (group C, 12), a group drawn immediately after exercise(group EI, 12), a group drawn 24 h after exercise(group EA, 12). Group C does not train. The rats in group EI and EA train on the treadmill with an increasing load for 6 weeks(10% grade, 6 d/w): in the first week, the rats run for 10 min at 10 m/min. Starting from the second week, the running speed increases by 5m/min/w, and the training time increases by 30min/w. In the last week the rats run to exhaustion if they could not maintain the target intensity. Record the exhausting time of rats, then group EI and group EA are respectively drawn immediately and 24 hours after exercise. Detect the apoptosis of renal cell apoptosis in rats by TUNEL, observe the ultrastructure of podocytes by TEM, detect urine total protein by BCA, serum and urine creatinine by Jaffe, serum urea by two-point dynamic method, the expression and distribution of Bcl-2 and Bax by immunohistochemistry, and the expression of Caspase-3 by western-blot. Results The rats in group EI and EA gradually lose weight at the first weekend of training, and their weight drop significantly from the third weekend to the end, it shows a significant difference compared with group C(p 0.05), while group EA is higher than group C and EI(p<0.05). Conclusions Persistent proteinuria is observed in rats after intensitive training, their renal function is disordered and cannot be effectively recovered after 24 h rest, and renal cell apoptosis increases. High-intensitive training reduces the expression of Bcl-2, and the Bax/Bcl-2 ratio increases with the prolongation of recovery time. Caspase-3 shows the same trend. It is suggested that the change in the expression and ratio of Bax and Bcl-2 in renal under excessive training stress is an important regulatory mechanism for the occurrence of apoptosis in renal cells. Meanwhile the apoptosis of renal cells increases in rats after training, and the apoptosis characteristics of podocyte in the glomerulus are obvious at 30 min and 24 h after exercise, and the structure and function of Slit diaphragm are damaged.

PO-081 Overtraining results in abnormality of renal silt diaphragm in rats with persistent proteinuria

Objective Silt diaphragm is the most important and bioactive membrane structure in the filtration barrier of kidney, and the root cause of proteinuria is the structural and functional abnormalities of Silt diaphragm. So far, there is little literature on the changes of silt diaphragm caused by overtraining. This research establishes a model of rats with exercise-induced proteinuria with long-term intensitive treadmill exercise, and it simulates the progressive-load training in the cycle of athletes. Histological and ultrastructural changes of kidney immediately and 24 h after exercise are observed, and it aims to analyze the change law of silt diaphragm during the occurrence of persistent proteinuria.&#x0D; Methods this study selects 36 Sprague-Dawley rats, which are randomly divided into 3 groups: a control group (group C, 12), a group drawn immediately after exercise(group EI, 12), a group drawn 24 h after exercise(group EA, 12). Group C does not train. The rats in group EI and EA train on the treadmill with an increasing load for 6 weeks(10% grade, 6 d/w): in the first week, the rats run for 10 min at 10 m/min. Starting from the second week, the running speed increases by 5m/min/w, and the training time increases by 30min/w. In the last week the rats run to exhaustion if they could not maintain the target intensity. Record the exhausting time of rats, then group EI and group EA are respectively drawn immediately and 24 hours after exercise. Observe the histological changes of renal glomerulus by optical microscope, and the ultrastructure of silt diaphragm by TEM. Detect urine total protein by BCA, serum and urine creatinine by Jaffe, serum testosterone and corticosterone by radioimmunoassay, serum urea by two-point dynamic method, and the expression of Nephrin by western-blot.&#x0D; Results The rats in group EI and EA gradually lose weight at the first weekend of training, and their weight drop significantly from the third weekend to the end, it shows a significant difference compared with group C(p&lt;0.01). There is no significant difference between the exercise group. Glomerular morphology, group C: The structure of glomerulus is compact, the boundary between vascular sphere and the wall of capsule is obvious, and the distribution of erythrocytes in vessels is regular; Group EI: The thickness of glomerulus membrane is uneven, the structure of the podocyte is incomplete, part of the foot process is fused, and SD is discontinuous; Group EA: Part of the glomerular endodermis is abnormal, part of the foot process is fused, detached, and unevenly distributed, and SD is discontinuous. Total protein/ creatinine in urine of rats 30 min and 24 h after exercise is significantly higher than that of group C(p&lt;0.01), and group EA is slightly retuned and lower than group EI(p&lt;0.05). Compared with group C, Serum Testosterone/Corticosterone of rats in group EI and EA is significantly decreased, and there is a significant difference (p&lt;0.01). However, there is a significant decrease in EA group but it is still significantly lower than group EI(p&lt;0.01). Serum corticosterone is significanty decreased in group EI and EA, and there is a significant difference (p&lt;0.01), while group EA is significantly decreased but still significantly lower than that in group EI(p&lt;0.01). Serum urea and espression of Nephrin are significantly decreased in group EI and EA(p&lt;0.01), but there are no significant difference between group EI and EA(p&gt;0.05).&#x0D; Conclusions The rats suffer from overtraining syndrome caused by intensitive training, and with persistent proteinuria, their renal function is disordered and cannot be effectively recovered after 24 h rest. Meanwhile, the renal morphology and ultrastructure of silt diaphragm in rats undergo "pathogenical-like" changes, which do not significantly decrease with the extension of recovery time. It is revealed that the injury of renal structure and ultrastructure of silt diaphragm caused by overtraining are the structural basis of continuous exercise-induced proteinuria.

Effect of emulsified isoflurane post-conditioning on mitochondrial function during lung ischemia-reperfusion in rats: an in vitro experiment

Objective To evaluate the effect of emulsified isoflurane post-conditioning on the mitochondrial function during lung ischemia-reperfusion (I/R) in rats in an in vitro experiment. Methods Twenty-four SPF healthy male Sprague-Dawley rats, weighing 250-300 g, were used in the study.After the animals were anesthetized, the lungs were removed, connected to the perfusion system and then divided into 4 groups (n=6 each) using a random number table: control group (group C), group I/R, emulsified isoflurane post-conditioning group (group EI) and intralipid post-conditioning group (group IL). After 20 min of equilibration, the lungs were continuously perfused for 105 min in group C, and the lungs were subjected to 45 min ischemia followed by 60 min reperfusion to establish the model of lung I/R injury in the other three groups.During the reperfusion period, the common perfusate was used in group I/R, the perfusate containing 1.68 mmol/L emulsified isoflurane was used in group EI, and the equal volume of perfusate containing 30% intralipid was used in group IL.At the end of the equilibration (T0), immediately after beginning of reperfusion (T1) and at 30 and 60 min of reperfusion (T2, 3), the arterial oxygen partial pressure (PaO2), airway resistance, pulmonary compliance and tidal volume (VT) were recorded.The right upper lobe of the lung was removed at T3 for determination of wet to dry weight ratio (W/D ratio). The right middle lobe of the lung was removed at T3 for pathologic examination with light microscope.The contents of reactive oxygen species (ROS), NAD+ and ATP in lung tissues were detected. Results Compared with group C, the PaO2, pulmonary compliance and VT were significantly decreased, and the airway resistance was increased at T1-3, and the W/D ratio and ROS content were increased, and NAD+ and ATP contents were decreased at T3 in I/R, EI and IL groups (P<0.05). Compared with I/R and IL groups, the PaO2, pulmonary compliance and VT were significantly increased, and the airway resistance was decreased at T2, 3, and the W/D ratio and ROS content were decreased, and NAD+ and ATP contents were increased at T3 in group EI (P<0.05). The pathologic changes of lungs were significantly attenuated in group EI as compared with group I/R. Conclusion The mechanism by which emulsified isoflurane post-conditioning attenuates lung I/R injury is related to decrease in mitochondrial dysfunction in rats in an in vitro experiment. Key words: Isoflurane; Fat emulsions, intravenous; Reperfusion injury; Lung; Mitochondrial; Ischemia post-conditioning

The predictive factors of α1-D/A adrenoceptor antagonist, naftopidil, dose increase therapy for male lower urinary tract symptoms caused by benign prostatic hyperplasia: INFORM study.

Introduction:We evaluated the predictive factors which affect the efficacy of naftopidil 50 mg/day therapy and dose increase therapy to administration of 75 mg/day after an initial dose of 50 mg/day.Materials and Methods:A total of 92 patients with male lower urinary tract symptoms/benign prostatic hyperplasia were administrated naftopidil 50 mg/day for 4 weeks (50 mg therapy). At week 4, the patients were divided into an effective and an ineffective group (Group E and Group I, respectively). For further 4 weeks, the dosage of naftopidil was increased to 75 mg/day in all patients. At week 8, the patients of Group E and Group I were divided into an effective and an ineffective group (Group EE, Group EI, Group IE, and Group II, respectively).Results:Postvoid residual (PVR) urine volume at baseline was a predictive factor for efficacy of 50 mg therapy. In Group E, change in International Prostate Symptom Score storage symptoms subscore from baseline to week 4 was a predictive factor for efficacy of this dose increase therapy. In Group I, change in maximum flow rate from baseline to week 4 was a predictive factor for efficacy of this dose increase therapy.Conclusions:The short term of naftopidil 50 mg therapy was ineffective for the patients who had large PVR. The predictive factor of this dose increase therapy might be a dynamic variable in 50 mg/day of dose period, but not a baseline variable at the time of 75 mg/day dosage starts.

Theoretical determination of aqueous acid–base pK values: electronic structure calculations and steered molecular dynamic simulations

The equilibrium constant (K) of several acid–base equilibria involving isomers of acetohydroxamic acid in aqueous solution is studied from a theoretical point of view applying electronic structure methods (at the M06-2X-SMD/6-311++G(d,p) and MP2-PCM/6-311++G(d,p) levels of theory) and steered molecular dynamic (SMD) simulations. The similarity of the results obtained indicates that SMD simulations can be successfully used to evaluate Gibbs energy changes in acid–base reactions in solution and pK values since these properties are in agreement with those found with quantum calculation and thermodynamic cycles. In the process of proton transfer from the imide isomers (the ZI and EI structures) toward the anion Z-amide, the deprotonation of the ZI(COH) molecule is the most favorable process kinetically and thermodynamically. It is observed that pK values are slightly higher for E-isomers and, particularly, for the deprotonation from the oxime group EI(NOH). Finally, we must emphasize the goodness of SMD simulations in solution to calculate this property as an alternative to using continuum solvation methods.

Effects of estrogen on epidermis growth of mice and proliferation of human epidermal cell line HaCaT and its mechanism

To observe the effects of estrogen on epidermis growth of mice and proliferation of keratinocytes (human epidermal cell line HaCaT), and to explore its mechanism. (1) Five adult C57BL/6 mice in estrus cycle were identified by vaginal exfoliative cytology diagnosis and set as estrus group, while another 5 adult C57BL/6 mice with ovary resected before sexual development were set as ovariectomized group. The full-thickness skin from the tail root of mice in two groups were collected. The thickness of epidermis was observed and measured after HE staining. The distribution of proliferating cell nuclear antigen (PCNA)-positive cells in epidermis was observed by immunohistochemical staining, the number of which was counted. (2) HaCaT cells in logarithmic growth phase were cultured with RPMI 1640 nutrient solution containing 10% fetal bovine serum, and they were divided into negative control group (NC), pure estradiol group (PE), protein kinase B (Akt) inhibitor group (AI), and extracellular signal-regulated kinase (ERK) inhibitor group (EI) according to the random number table, with 20 wells in each group. To nutrient solution of each group, 1 μL dimethyl sulfoxide, 1 μL 17β-estradiol (100 nmol/L), 1 μL LY294002 (10 μmol/L), and 1 μL PD98059 (30 μmol/L) were added in group NC, group PE, group AI, and group EI respectively, and the last two groups were added with 1 μL 17β-estradiol (100 nmol/L) in addition. At post culture hour (PCH) 0 (immediately after culture), 24, 48, 72, 5 wells of cells from each group were collected to detect the proliferation activity of cells by cell counting kit 8 and microplate reader. (3) HaCaT cells in logarithmic growth phase were collected, grouped, and treated with the above-mentioned methods, with 3 wells in each group. At PCH 72, cell cycle distribution was detected by flow cytometer to calculate proliferation index (PI) of cells. (4) HaCaT cells in logarithmic growth phase were collected, grouped, and treated with the above-mentioned methods, with 3 dishes in each group. At PCH 72, the protein levels of phosphorylated Akt (p-Akt), phosphorylated ERK (p-ERK), and PCNA were determined with Western blotting. The cell experiments were repeated for 3 times. Data were processed with t test, one-way analysis of variance, analysis of variance of factorial design, and LSD test. (1) The epidermis thickness of mice in ovariectomized group was (33.5±3.0) μm, which was obviously thinner than that in estrus group [(51.4±3.1) μm, t=20.7, P<0.01]. The PCNA-positive cells mainly aggregated in the basal layer of epidermis of mice in two groups. The number of PCNA-positive cells in epidermis of mice in ovariectomized group was 37±12 per 200 fold visual field, obviously fewer than that in estrus group (96±15 per 200 fold visual field, t=15.3, P<0.01). (2) During PCH 0 to 48, there were no significant differences in the proliferation activity of cells between group PE and group NC (with P values above 0.05). At PCH 72, compared with that in group NC, the proliferation activity of cells in group PE was obviously increased (P<0.01). The proliferation activity of cells in groups AI and EI was obviously lower than that in the previous two groups (with P values below 0.01). (3) Compared with that in group NC [(51.6±1.1)%], the PI of cells in group PE was obviously increased [(58.5±0.8)%, P<0.05]. The PI values of cells in groups AI and EI were (34.9±0.8)% and (48.2±0.4)% respectively, both obviously lower than those in the previous two groups (with P values below 0.01). (4) Compared with that of group NC (0.566±0.034), the protein level of p-Akt in cells of group PE was significantly increased (1.048±0.077, P<0.01). Compared with that of group PE, the protein level of p-Akt was obviously decreased in cells of groups AI and EI (respectively 0.682±0.095 and 0.672±0.019, with P values below 0.01). Compared with that of group NC (0.469±0.013), the protein level of p-ERK obviously increased in cells of groups PE, AI, and EI (respectively 1.064±0.089, 1.010±0.038, 0.778±0.065, with P values below 0.01). The protein level of p-ERK in cells of group EI was obviously lower than that in group PE (P<0.01). Compared with that of group NC (0.386±0.053), the protein level of PCNA was obviously increased in cells of group PE (0.743±0.043, P<0.01). The protein levels of PCNA in cells of groups AI and EI were 0.264±0.019 and 0.223±0.065 respectively, both obviously lower than those in the previous two groups (with P values below 0.01). Lack of estrogen damages the growth ability of epidermis of mice. Estrogen (17β-estradiol) can promote the proliferation of HaCaT cells by increasing the expression of PCNA via activating ERK/Akt signaling pathway.

Comparison of the effects of propofol and emulsified isoflurane alone or combined with dexmedetomidine on induction of anesthesia in dogs

ObjectiveTo compare the respective effects of propofol and emulsified isoflurane administered alone and in combination with dexmedetomidine on the quality of induction of anesthesia, physiological variables and recovery in dogs. Study designProspective, randomized, experimental trial. AnimalsThirty-six adult mixed-breed dogs. MethodsAnimals were randomly assigned to one of four induction protocols: propofol alone (group P); emulsified isoflurane alone (group EI); both propofol and dexmedetomidine (group PD), or both emulsified isoflurane and dexmedetomidine (group EID). Pulse rate (PR), respiratory rate (fR), non-invasive arterial blood pressure and arterial blood gases were measured at baseline, before induction, immediately after intubation (time 0), and at 5 minute intervals until the dog began to swallow and the trachea was extubated. The quality of induction and recovery, and degree of ataxia were scored by a single investigator unaware of group assignment. The durations of anesthesia and recovery, and the incidence of adverse events were recorded. ResultsThere were no clinically significant differences among the groups in induction quality. Systolic arterial pressure was lower in EID compared with P at 5 minutes. PR and fR were lower in PD and EID compared with P after induction. The PaCO2 at 5 minutes was higher than at baseline in group P. Ataxia score was lower in EID than in P. Time from induction to extubation and time from extubation to sternal recumbency were lower in EID compared with PD. Conclusions and clinical relevanceThere were no clinically significant differences among the groups in induction quality. In PD and EID, but not in P, PR and fR were decreased after induction. The EID combination resulted in smooth and rapid induction and recovery and thus may be useful clinically for induction of anesthesia.

Effects of emulsified isoflurane preconditioning on lung ischemia-reperfusion injury in rats

Objective To investigate the effects of emulsified isoflurane preconditioning on lung ischemia-reperfusion(I/R)injury in rats. Methods Thirty-two male Sprague-Dawley rats, weighing 250-320 g, were randomly divided into 4 groups(n=8 each)using a random number table: sham operation group(group S), group I/R, intralipid group(group IL), and emulsified isoflurane preconditioning group(group EI). Lung I/R was produced by clamping the left hilum of lungs for 1 h followed by 2 h of reperfusion in anesthetized rats.In group S, intraperitoneal normal saline 10.5 ml/kg was injected, but the hilum was only exposed 24 h later.In groups I/R, IL and EI, intraperitoneal normal saline 10.5 ml/kg, 30% intralipid 10.5 ml/kg and 8% emulsified isoflurane 10.5 ml/kg were injected intraperitoneally, respectively, and then the model was established 24 h later.At the end of reperfusion, blood samples were collected from the left ventricle for measurement of PaO2, PaCO2, plasma malondialdehyde(MDA)concentration(by ELISA). The rats were then sacrificed, and left lungs were removed immediately for microscopic examination and for determination of myleoperoxidase(MPO)activity(using colorimetric method)in pulmonary specimens.Wet/dry lung weight ratio(W/D ratio)was calculated. Results Compared with group S, W/D ratio, MPO activity, plasma MDA concentration and PaCO2 were significantly increased, and PaO2 was decreased in the other three groups.Compared with group I/R, W/D ratio, MPO activity, plasma MDA concentration and PaCO2 were significantly decreased, and PaO2 was increased in IL and EI groups.W/D ratio, MPO activity, plasma MDA concentration and PaCO2 were significantly lower, and PaO2 was higher in group EI than in group IL.The pathological changes of the lung were significantly attenuated in group EI compared with group I/R. Conclusion Emulsified isoflurane preconditioning can reduce lung I/R injury in rats, and inhibition of pulmonary inflammatory responses and of production of oxygen free radicals are involved in the mechanism. Key words: Isoflurane; Fat emulsions, intravenous; Ischemic preconditioning; Reperfusion injury; Lung

Multiple Levels Analysis of Group EI Infecting Employee EI and Performance

On the one hand, as the basic work forms, team is paid more and more attention. Team is a important view of research. At same time, studies find that emotional intelligence is not only a tool in management, but also is antecedent variables. But, on the other hand, according to the previous study, the researches on relation between emotional intelligence and performances are limited in individual level, and don’t make a deep study to the mechanism between them. Based on the two points, this paper chooses team members as object of study, and researches the mechanism from individual level and team level. These results open the black box that emotion intelligence effects performance. At same time, these results provide a good research perspective and bases for follow-up research and in human resource management practices.

The effect of experimental exposure to low doses of zearalenone on uterine histology and morphometry in prepubertal bitches

The experiment involved 30 clinically healthy prepubertal bitches aged approximately 70 days with an estimated initial body weight (BW) of 8 kg. The animals were randomly divided into two experimental groups (EI and EII) and a control group of 10 animals each. Group EI was administered 50 μg zearalenone (ZEN)/kg BW per os for 42 days, group EII received 75 μg zearalenone/kg BW per os for 42 days, and the control group was administered placebo per os for 42 days. The bitches were hysterectomized at the end of treatment, and samples of uterine tissue were collected for histological and morphometric analyses. The results of the study indicate that exposure to very low doses of ZEN (100% and 150% of the NOAEL) causes simple glandular hyperplasia of the endometrium accompanied by adenogenesis, angiogenesis, and vasodilatation with the related consequences. The noted changes were more pronounced in group EI and less visible in group EII in comparison with group C, which could be indicative of a hormetic dose response.

The effect of low doses of zearalenone and its metabolites on progesterone and 17β-estradiol concentrations in peripheral blood and body weights of pre-pubertal female Beagle dogs

The experiment involved 30 clinically healthy female Beagle dogs aged approximately 70 days with estimated initial body weight (BW) of 8 kg. The animals were randomly divided into two experimental groups (EI and EII) and a control group of 10 animals each. Group EI was intoxicated with 50 μg zearalenone/kg BW per os for 42 days, group EII received 75 μg zearalenone/kg BW per os for 42 days, and the control group was administered placebo per os for 42 days. The animals were weighed, and blood samples for analyses of the concentrations of zearalenone, its metabolites, progesterone and 17β-estradiol were collected seven times at seven-day intervals, one hour after mycotoxin administration. Biotransformation of zearalenone was observed in all groups throughout the experiment, and the highest percentage share of α-zearalenol was reported in group EII on the last five sampling dates (0.637–0.788 ng/ml, i.e. percentage share of 57.96–73.64%). The above had a significant influence on the non-physiological concentrations of progesterone and 17β-estradiol in both experimental (E) groups throughout the experiment. The lowest progesterone levels (0.131 ng/ml) were observed in group EII during the last test, and high concentrations of 17β-estradiol were found in group EII on the last two sampling dates (17.434 and 21.581 ng/ml, respectively) in comparison with control. Inhibited proliferation, manifested by a slower rate of body weight gain, was observed on the last but one day of zearalenone administration in both experimental groups. Our results indicate that NOAEL doses have stimulating/adaptive effects, whereas doses above NOAEL values suggest that even very low zearalenone doses can act as endocrine disruptors with regard to progesterone and 17β-estradiol.

The effects of experimental administration of low doses of zearalenone on the histology of ovaries in pre-pubertal bitches

The experiment involved 30 clinically healthy Beagle bitches aged approximately 70 days with an initial body weight of approximately 8 kg. The animals were randomly divided into two experimental groups (EI and EII) and a control group of 10 animals each. Group EI was administered 50 microg of body weight zearalenone/kg per os for 42 days, group EII received 75 microg of body weight zearalenone/kg per os for 42 days, and the control group was administered placeboper os for 42 days. The bitches were ovariectomized at the end of the treatment period for anatomopathological examination. At the same time, peripheral blood samples were collected for endocrinological analyses (17beta-estradiol and progesterone). Administartion of zearalenone particularly higher doses, resulted in the hyperestrogenism degeneration and atrophy of ovarian cells and tissues with accompanying edema and blood extravasation, leading to increased 17beta-estradiol concentrations and an insignificant decrease in progesterone levels.

Abstract 1873: Effect of intermittent calorie restriction on cancer stem cells.

Abstract Effect of intermittent calorie restriction (ICR) on cancer is controversial. We then examined the effect of ICR and food contents on cancer using mouse model. CT26 mouse colon cancer cells were inoculated into the back of syngeneic BALB/c mouse. A 24 hours-fasting was performed once a week for 4 weeks. A control diet and a high calorie diet were used for feeding. The body weight at week 4 of mice was not different among control diet (Group C), control diet+ICR (Group CI) and high fat diet+ICR (Group FI). The tumor weight, nodal metastasis and CD133-positive cancer stem cells (CSC) in the tumor were more pronounced in Group CI and FI than those in Group C and F. Blood CD133 mRNA levels after fasting breakage were increased in Group CI and FI. We have reported the prometastatic effect of trans fatty acids in colorectal cancer. Here effect of oral intake of trans fatty acids on tumor stem cells with ICR was examined using the same model. Mice fed with 10% elaidic acid diet with ICR- (Group E) or ICR+ (Group EI) were compared with Group C and CI. The tumor weight and tumor MIB1 index was Group EI, Group E, Group CI and Group C in high order. In Group EI, nucleostemin (NS)-positive tumor cells were transiently increased on the next day of ICR. During 12 ICR cycle, NS-positive CSCs were gradually increased in Group EI. These findings suggest that ICR might enhance cancer stem cell-activating factors. Citation Format: Yi Luo, Yasuhiko Kitadai, Yoshitomo Chihara, Rina Fujiwara, Kiyohide Fujimoto, Hiroki Kuniyasu. Effect of intermittent calorie restriction on cancer stem cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1873. doi:10.1158/1538-7445.AM2013-1873

The effects of joint immobilization on articular cartilage of the knee in previously exercised rats

Studies have determined the effects of joint immobilization on the articular cartilage of sedentary animals, but we are not aware of any studies reporting the effects of joint immobilization in previously trained animals. The objective of the present study was to determine whether exercise could prevent degeneration of the articular cartilage that accompanies joint immobilization. We used light microscopy to study the thickness, cell density, nuclear size, and collagen density of articular cartilage of the femoral condyle of Wistar rats subjected to aerobic physical activity on an adapted treadmill five times per week. Four groups of Wistar rats were used: a control group (C), an immobilized group (I), an exercised group (E), and an exercised and then immobilized group (EI). The right knee joints from rats in groups I and EI were immobilized at 90 °C of flexion using a plastic cast for 8 weeks. Cartilage thickness decreased significantly in group I (mean, 120.14 ± 15.6 μm, P < 0.05), but not in group EI (mean, 174 ± 2.25), and increased significantly in group E (mean, 289.49 ± 9.15) compared with group C (mean, 239.20 ± 6.25). The same results were obtained for cell density, nuclear size, and collagen density (in all cases, P < 0.05). We concluded that exercise can prevent degenerative changes in femoral articular cartilage caused by immobilization of the knee joint.