Imaging characterization of myocardial function, fibrosis, and perfusion in a nonhuman primate model with heart failure-like features.

Introduction: The availability of a human-like chronic heart failure (HF) animal model was critical for affiliating development of novel therapeutic drug treatments. With the close physiology relatedness to humans, the non-human primate (NHP) HF model would be valuable to better understand the pathophysiology and pharmacology of HF. The purpose of this work was to present preliminary cardiac image findings using echocardiography and cardiovascular magnetic resonance (CMR) in a HF-like cynomolgus macaque model. Methods: The NHP diet-induced model developed cardiac phenotypes that exhibited diastolic dysfunction with reduced left ventricular ejection fraction (LVEF) or preserved LVEF. Twenty cynomolgus monkeys with cardiac dysfunction were selected by echocardiography and subsequently separated into two groups, LVEF < 65% (termed as HFrEF, n = 10) and LVEF ≥ 65% with diastolic dysfunction (termed as HFpEF, n = 10). Another group of ten healthy monkeys was used as the healthy control. All monkeys underwent a CMR study to measure global longitudinal strain (GLS), myocardial extracellular volume (ECV), and late gadolinium enhancement (LGE). In healthy controls and HFpEF group, quantitative perfusion imaging scans at rest and under dobutamine stress were performed and myocardial perfusion reserve (MPR) was subsequently obtained. Results: No LGE was observed in any monkey. Monkeys with HF-like features were significantly older, compared to the healthy control group. There were significant differences among the three groups in ECV (20.79 ± 3.65% in healthy controls; 27.06 ± 3.37% in HFpEF group, and 31.11 ± 4.50% in HFrEFgroup, p < 0.001), as well as for stress perfusion (2.40 ± 0.34 ml/min/g in healthy controls vs. 1.28 ± 0.24 ml/min/g in HFpEF group, p < 0.01) and corresponding MPR (1.83 ± 0.3 vs. 1.35 ± 0.29, p < 0.01). After adjusting for age, ECV (p = 0.01) and MPR (p = 0.048) still showed significant differences among the three groups. Conclusion: Our preliminary imaging findings demonstrated cardiac dysfunction, elevated ECV, and/or reduced MPR in this HF-like NHP model. This pilot study laid the foundation for further mechanistic research and the development of a drug testing platform for distinct HF pathophysiology.

Clinical study of invasive pulmonary aspergillosis following influenza A H1N1.

ABSTRACT: This study to analyze the clinical characteristics of patients with invasive pulmonary aspergillosis (IPA) following influenza A (H1N1) infection.We retrospectively analyzed 10 cases with IPA following H1N1 infection. The clinical manifestations, laboratory examination results, chest computed tomography, and treatments were analyzed.Clinical manifestations: all 10 cases had typical flu-like symptoms at the onset of the disease, among which 7 patients developed dyspnea in the late stage, and 8 patients had hemoptysis. Laboratory examination: the absolute and percentage of peripheral blood lymphocytes in all 10 patients were declined, among which 5 cases were with decreased CD3+ CD4+ T cells/lymphocytes; 9 cases with increased bronchoalveolar lavage fluid galactomannan; 6 cases with increased serum galactomannan; 1 case with bronchoalveolar lavage fluid cultured aspergillus fumigatus; and 2 cases with aspergillus by second-generation sequencing. Chest computed tomography: all patients showed multiple diffused ground-glass opacities at the beginning, along with linear or reticular interstitial changes. Two cases had multiple subarachnoid nodules with halo signs, 3 cases had consolidation in multiple segments of both lungs, 2 cases had cavities, and 4 cases were with pleural effusion. Treatment: 10 patients were treated with antiviral and anti-Aspergillus drugs after admission. Four patients received respiratory support. All 10 cases were cured and discharged.Early diagnosis of IPA in influenza A (H1N1) patients is the key to successful treatment.

Prevalence and determinants of potentially inappropriate medications prescribing in elderly patients in Chinese communities.

BACKGROUND: To assess the prevalence and associated risk of potentially inappropriate medications (PIMs) prescribing in community-dwelling elderly patients in China and to examine the most frequently used PIMs. This will provide a reference for the formulation of medication manuals for the community-dwelling elderly and further standardize the use of medications in elderly patients. METHODS: We conducted a cross-sectional retrospective study from April 1, 2020 to April 30, 2020. Data from elderly patients aged ≥65 years were collected from the Hengjie (N=2,294), Loujiang (N=3,972), and Tongxing communities (N=1,969) in Suzhou. The frequency of PIMs was detected using the 2019 Beers criteria and the 2017 Chinese criteria. Chi-square (for categorical variables), ANOVA (for continuous variables as applicable), and logistic regression were used to describe and identify potential predictors of PIMs. RESULTS: A total of 8,235 elderly patients were examined. Using the Chinese criteria, the prevalence of PIMs was 37.07%, which was slightly higher than that found using the 2019 Beers criteria (32.16%). The most prescribed PIMs were estazolam (21.53%) and insulin (4.60%) based on the Chinese criteria. Logistic regression analysis showed that advanced age, polypharmacy, and comorbid disease of patients were associated with a high risk of PIMs. Furthermore, the educational background and professional title of physicians were also associated with PIMs. CONCLUSIONS: Given the high prevalence of PIMs in the Chinese community-dwelling elderly population, the implementation of evidence-based interventions to promote rational clinical drug use could improve their quality of life.

Pathological study of otitis media with effusion after treatment with intranasal pulmonary surfactant.

OBJECTIVES/HYPOTHESIS: To evaluate the histopathological effect of intranasal pulmonary surfactant (PS) on the eustachian tube (ET) in guinea pigs with otitis media with effusion (OME). STUDY DESIGN: Randomized control trial. METHODS: Nonviable heat-killed Hemophilus influenzae solution was injected into the tympanum of guinea pigs by a trans-eardrum approach to establish OME. Guinea pigs were divided into four groups: normal controls (group A), untreated OME (group B), saline-treated (group C), PS-treated (group D). The response threshold of the guinea pigs was measured by auditory brainstem response (ABR), and data were analyzed by one-way analysis of variance. The histopathological changes in the osseous, cartilaginous, and muscular portions of the ET were observed systematically by light microscopy. RESULTS: The ABR threshold in OME group B was raised significantly compared with normal group (A). The response in saline-treated group C was not statistically significantly different compared with OME group B. Seven days after intranasal dripping of pulmonary surfactant in PS-treated group D, the response threshold showed at statistically significant decrease compared with OME B and saline-treated C groups. In OME group B and saline-treated group C, mucosa showed swelling with goblet cell hyperplasia, and cilia were irregularly arranged. In PS-treated group D, there was slight mucosal swelling with fewer goblet cells, and cilia were regularly arranged, similar to the normal group A. CONCLUSIONS: The findings of the study indicate that intranasal pulmonary surfactant drops have protective and hyposecretory effects on the mucociliary system of the ET in guinea pigs suffering from OME.

Relationship between symptoms and weight-bearing radiographic parameters of idiopathic flexible flatfoot in children.

BACKGROUND: Idiopathic flexible flatfoot in children most frequently improves with age and remains asymptomatic. It is a physiological variation of the normality that does not require treatment unless it becomes symptomatic. The aim of this research was to investigate the reason why some individuals with flexible flatfoot become symptomatic by analysis of the differences in the relative alignment of each segment of the foot between symptomatic and asymptomatic patients with idiopathic flexible flatfoot using radiographic measurements. METHODS: One hundred patients with idiopathic flexible flatfoot were retrospectively identified and divided into two groups: asymptomatic (n = 50) and symptomatic (n = 50). Standing anteroposterior and lateral radiographs of the foot were analyzed. Five measurements were calculated to describe the alignment of the foot. An independent-samples t-test and Logistic regression test were used for statistical analysis. RESULTS: Age and sex were similar in the two groups. The independent-samples test revealed significant differences in two parameters: the anteroposterior talonavicular coverage angle and the lateral talo-first metatarsal angle. When the Logistic regression test was performed, only the talonavicular coverage angle showed statistical significance. CONCLUSIONS: The lateral displacement of the navicular bone, measured by the anteroposterior talonavicular coverage angle, seems to be related to the onset of symptoms. In individuals with otherwise normal flexible flatfoot, an increase in this angle might be an important risk factor for developing symptoms.

Limb remote ischemic preconditioning protects the spinal cord from ischemia-reperfusion injury: a newly identified nonneuronal but reactive oxygen species-dependent pathway.

BACKGROUND: It remains to be established whether spinal cord ischemic tolerance can be induced by limb remote ischemic preconditioning (RIPC), and the mechanisms underlying the neuroprotective effects of RIPC on the spinal cord need to be clarified. METHODS: Spinal cord ischemia was studied in New Zealand White rabbits. In experiment 1, all rabbits were subjected to 20-min spinal cord ischemia by aortic occlusion. Thirty minutes before ischemia, rabbits were subjected to sham intervention or RIPC achieved by bilateral femoral artery occlusion (10 min ischemia/10 min reperfusion, two cycles). Dimethylthiourea (500 mg/kg, intravenously), a hydroxyl radical scavenger, or vehicle was given 1 h before RIPC. Antioxidant enzyme activity was measured along with spinal cord histology and neurologic function. In experiment 2, rabbits were subjected to spinal cord ischemia, with or without RIPC. In addition, rabbits were pretreated with various doses of hexamethonium. RESULTS: RIPC improved neurologic function and reduced histologic damage. This was associated with increased endogenous antioxidant activity. Dimethylthiourea inhibited the protective effects of RIPC. In contrast, there was no effect of hexamethonium on the protective effect of RIPC. CONCLUSIONS: An initial oxidative stress acts as a trigger to upregulate antioxidant enzyme activity, rather than the neural pathway, and plays an important role in the formation of the tolerance against spinal cord ischemia by limb RIPC.

Therapeutic time window and mechanism of tetramethylpyrazine on transient focal cerebral ischemia/reperfusion injury in rats.

The purpose of this study was to explore the therapeutic time window and mechanism of tetramethylpyrazine on transient focal cerebral ischemia/reperfusion injury. Middle cerebral artery occlusion was conducted in male Sprague-Dawley rats and 20mg/kg tetramethylpyrazine was injected intraperitoneally at different time points. Neurological deficit scores and brain infarction volumes were measured 72 h after reperfusion started. The expression of thioredoxin and thioredoxin reductase were examined at 6h and at 24h after reperfusion. Our results included the findings of a significant reduction in neurological deficit scores and infarction volume in the treatment group as compared to the control group. Ischemia/reperfusion injury resulted in a decrease in the expression of thioredoxin, while tetramethylpyrazine administration greatly elevated the expression of thioredoxin-1/thioredoxin-2 mRNA and thioredoxin reductase-1/thioredoxin reductase-2 mRNA. These findings suggest that administration of tetramethylpyrazine, within a 4h time period post-transient focal stroke, may reduce cerebral ischemic reperfusion damage. Moreover, the neuroprotective effect of tetramethylpyrazine may be mediated, in part, by an increase in genetic transcription of thioredoxin.

[Neuroprotective effect of preconditioning with cannabinoid receptor agonist WIN 55, 212 - 2 on focal cerebral ischemia: experiment with rats].

OBJECTIVE: To investigate the neuroprotective effect of preconditioning with cannabinoid (CB) receptor agonist WIN 55, 212 - 2 on focal cerebral ischemia. METHODS: Fifty male SD rats were randomly assigned to 5 equal groups: control group undergoing middle cerebral artery occlusion (MCAO) for 2 h only without any preconditioning; 3 WIN 55, 212 - 2 preconditioning groups (WIN1-3) injected intraperitoneally with WIN 55, 212 - 2 at the doses of 0.3, 1, and 3 mg/kg respectively 24 h before MCAO for 2 h, and DMSO group injected intraperitoneally with dimethyl sulfoxide (DMSO), solvent of WIN 55, 212 - 2 24 h before MCAO for 2 h. 24, 48, and 72 hours after reperfusion the neurological function score (NFS) was evaluated the rats were then decapitated with their brains taken out. Brain infarct volume was evaluated with 2% 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. RESULTS: The NFS values of the rats in WIN 55, 212 - 2 preconditioning groups were all significantly higher than those of the control and DMSO groups (all P < 0.05) while the infarct volumes of the WIN 55, 212 - 2 preconditioning groups were all significantly smaller than those of the control and DMSO groups (all P < 0.05) 24, 48, and 72 h after reperfusion. The infarct volumes of the WIN2 and WIN3 groups were both significantly smaller than that of the WIN1 group (both P < 0.05). However, there was no significant difference in the infarct volume between WIN2 and WIN3 groups (P = 0.928). CONCLUSION: WIN 55, 212 - 2 preconditioning has neuroprotective effect on focal cerebral ischemia with a dose-dependent manner.

[Effects of electroacupuncture at "Zusanli" (ST 36) on ERK1/2 phosphorylation in the dorsal horn of spinal cord of the rat].

OBJECTIVE: To observe the effect of electroacupuncture at Zusanli (ST 36) on phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) in the dorsal horn of spinal cord induced by plantar inflammation in the rat. METHODS: All the rats were randomly divided into 5 groups: normal control group, simple electroacupuncture group, formalin group, formalin plus ipsilateral electroacupuncture group and formalin plus contralateral electroacupuncture group. The acute inflammation animal model was made by injection of 100 microL of 4% formalin into the right posterior foot pad. Electroacupuncture was given at "Zusanli" (ST 36) for 30 min, with sparse-dense waves, frequency 2-15 Hz, and intensity 2-3 mA. One and a half hours latter, the rats were killed under anesthesia, and pERK1/2 expression in the lumbar dorsal horn were investigated with immunohistochemical method. RESULTS: The positive cells were rarely seen (6.45 +/- 1.05) in the superficial spinal cord in the control group; a few cells (14.07 +/- 3.19) in ipsilateral superficial spinal cord were found in the electroacupuncture group. The number of pERK1/2-positive neurons (26.57 +/- 4.93) in lamina I - II0 of the ipsilateral dorsal horn in the formalin group increased significantly. After electroacupuncture at ipsilateral Zusanli (ST 36), the number of positive cells (20.79 +/- 5.21) had a tendency to decrease, but with no statistically significant difference. However, after electroacupuncture at contralateral Zusanli (ST 36), the number of positive cells (14.75 +/- 3.03) significantly decreased as compared with the non-acupuncture group (P < 0.05). CONCLUSION: The inhibition of ERK1/2 phosphorylation in the spinal cord dorsal horn by electroacupuncture is possibly involved in acupuncture analgesic effect.

Effect of water restrictions on the physiological parameters, psychological behavior and brain c-Fos expression in rats.

OBJECTIVE: In order to characterize the feature of stress response induced by stressor with both physical and psychological natures, the effects of water restriction performed in different experimental modes on the physiological parameters, psychological behavioral manifestations and brain c-Fos expressions were observed and compared. METHODS: Fifty-eight male Wistar rats were used and randomly divided into three experimental groups (n = 18 for each) and a control group (n = 4). In control group, the rats were allowed to access drinking water freely at all experimental period. In the experimental groups the water supply to the rats was restricted. In timed water supply (TW) group, the water was supplied twice a day, 10 min for each in fixed hours every day. In empty bottle-served (EB) and water-restricted (WR) groups, the water was served only once a day for 10 min, either in the early morning or evening, and in the other time point scheduled for water supply only an empty bottle without water was provided in the EB group and nothing was given in the WR group. The quantities of drank water and eaten food, weight-gaining, and behavior score were observed every day. The serum level of corticosterone was assayed and the rats were sacrificed with fixative perfusion of 3 d, 7 d or 14 d, respectively, following water restriction (n = 6 for each time point in each group). The brain c-Fos expressions were examined with immunohistochemical method. RESULTS: The slow down of weight-gaining, rise of serum corticosterone level, occurrence of psychological behavioral manifestations of unpeaceful restlessness such as exploring and attacking, enhance of c-Fos expression in the subfornical organ (SFO), median preoptic nucleus (MnPO), area postrema (AP), hypothalamic paraventricular nucleus (PVN), supraoptic nucleus (SON), medial (MeA) and central (CeA) amygdaloid nucleus and ventrolateral septum (LSV) were noticed in both EB and WR groups, except the nucleus of solitary tract (NTS) in which the Fos expression was decreased. The changes of Fos expression in most of nuclei in EB group began at day 3, at least persisted till day 7, and backed down at day 14, while in WR group, similar changes started at day 7 and reached its peak at day 14. In TW group, only the concentration of corticosterone at day 7 was slightly increased and the rest indexes observed were unchanged. CONCLUSION: The results indicate that water restriction induces physical as well as psychological stress responses. And the water restrictions experimentally executed in different modes result in different manifestations of behavioral response and brain immediately early gene expression in discrete brain nuclei/regions.

[Preparation of polyclonal and monoclonal antibodies against CRF and change of CRF in the sleep-deprived rat brain].

AIM: To produce polyclonal and monoclonal antibodies against corticotropin-releasing factor (CRF) and to study the changes of CRF in sleep-deprived rat brain with the antibodies acquired. METHODS: Commercial CRF was linked to bovine throglobulin (BTG) and bovine serum albumin (BSA) respectively to produce immunogen and embedding antigen. New Zealand rabbits and BALB/c mice were immunized with the BTG-CRF immunogen to produce polyclonal and monoclonal antibodies, respectively. The acquired antibodies were appraised with ELISA and immnohistochemical staining. The characterized antibodies were used to observe the changes of CRF in the rat brain 48 h after sleep deprivation. RESULTS: CRF polyclonal antibody and 9 clones of monoclonal antibodies were obtained with high titer, affinity and specificity. Among them, the polyclonal antibody and 2 monoclonal antibodies (1D10, 2F4) were excellent in immunocytochemical staining. The CRF-like immunoreactive substances were found more strongly expressed in the neurons of paraverticular hypothalamic nucleus (PVN), central subnucleus of amygdala (CeA), oval subnucleus of bed nucleus of stria terminalis (BNSTov), and nucleus of solitary tract (NTS) in sleep-deprived rat brain. While they were much weaker and even absent in the control. CONCLUSION: The polyclonal and monoclonal antibodies against CRF were successfully produced for immunocytochemical studies. The results indicate that CRF may play an important role in stress-responsive modulation during sleep deprivation. PVN, CeA and BNSTov are integral part of brain circuit related to the stress modulation.

[Research of fibronectin, p53 and Ezrin protein with occurrence, progress and prognosis in the primary laryngeal carcinoma by the tissuechip technology].

OBJECTIVE: To evaluate the predictive role of fibronectin, p81 (Ezrin protein) and p53 gene in primary laryngeal carcinoma, it's relationship with epidemiology(smoking), histological grading, surgical treatment, TNM stage and prognosis were studied by the tissuechip technology. METHODS: The expression of fibronectin, p53 gene and p81 (Ezrin protein) on a series of 85 primary laryngeal carcinoma patients treated in our hospital between 1992 and 2000 was studied with tissuechip technology. The correlation of each score according to the intensity and percentage of labeled cells or intercellular substance with relevant clinical dada was statistically analyzed. RESULTS: Some cases were lost or boosted no tumor tissue in our tissuechip. Among the 70 cases available, 45.71% (32/70) of the specimens' basal membrane and extracellular matrix were strongly stained with fibronectin; there is statistical significance (P < 0.05) between primary tumor grading groups. Ezrin protein expressing rate is 87.3%, and the average percentage of its labeled cells is 53.68% (ranging from 0% to 100%, median is 58. 69%). There were significant difference between tumor grading groups, clinical early and late stages and 3-year survival rates (P < 0.05) after chi-square test. But no relation with smoking, gender, age and histological classes (P > 0.05). The average percentage of p53 positive cells is 21.6% (ranging from 0% to 90.3%, median is 5.85%) and 46.8% showed positive stains in our research. There was no statistical prominence in p53 protein demonstration between TNM stages, lymph node metastasis, 3-year survival rate, smoking, gender, age and histological classes (P > 0.05). CONCLUSIONS: The tissue microarray technique spent shorter time and less expense, and showed higher consistency in our essays. And the present study suggests fibronectin and p81 (Ezrin protein) could be the clinical discriminators in laryngeal carcinoma.